doc_id sent_index relation_id relation trigger trigger_offset arg_num arg_head arg_protein arg_domain arg_site arg_sugar arg_head_offset arg_base_np arg_base_np_offset arg_np arg_np_offset sent_text 22688517 5 0 gly glycopeptides 1059:1071 arg2 glycopeptides 1059:1071 glycopeptides 1059:1071 glycopeptides from non-specific protease digestion 1059:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 0 gly glycopeptides 1059:1071 arg1 digestion 1100:1108 non-specific protease digestion 1078:1108 non-specific protease digestion 1078:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 1 theme glycans 1452:1458 arg1 structure 1421:1429 the structure 1417:1429 the structure of glycopeptides and glycans 1417:1458 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 5 2 theme MS 683:684 arg1 analysis 694:701 MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 651:701 MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 651:701 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 3 theme spectra 1485:1491 arg1 interpretation 1493:1506 proper spectra interpretation 1478:1506 proper spectra interpretation 1478:1506 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 7 4 theme glycopeptides 1434:1446 arg1 structure 1421:1429 the structure 1417:1429 the structure of glycopeptides and glycans 1417:1458 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 6 5 theme type 1266:1269 arg1 N-glycans 1271:1279 heterogeneous complex type N-glycans 1244:1279 heterogeneous complex type N-glycans 1244:1279 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 5 6 from analysis 1047:1054 arg1 digestion 1100:1108 non-specific protease digestion 1078:1108 non-specific protease digestion 1078:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 7 with combination 798:808 arg1 confirmation 832:843 glycan structure confirmation 815:843 glycan structure confirmation 815:843 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 6 8 theme complex 1258:1264 arg1 N-glycans 1271:1279 heterogeneous complex type N-glycans 1244:1279 heterogeneous complex type N-glycans 1244:1279 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 5 9 theme released 868:875 arg1 glycans 877:883 released glycans 868:883 released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion 868:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 10 theme glycopeptides 1059:1071 arg1 analysis 1047:1054 direct MS analysis 1037:1054 direct MS analysis of glycopeptides from non-specific protease digestion 1037:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 2 11 theme LOX-1 235:239 arg1 N-glycosylation 216:230 The N-glycosylation 212:230 The N-glycosylation of LOX-1 212:239 The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. 22688517 4 12 theme present 427:433 arg1 study 435:439 The present study 423:439 The present study 423:439 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 5 13 theme non-specific 1078:1089 arg1 digestion 1100:1108 non-specific protease digestion 1078:1108 non-specific protease digestion 1078:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 2 14 from functions 281:289 arg1 vivo 294:297 vivo 294:297 vivo 294:297 The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. 22688517 0 15 theme low 87:89 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 15 theme low 87:89 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 4 16 gly N-glycosylation 549:563 arg2 sites 565:569 N-glycosylation sites 549:569 N-glycosylation sites 549:569 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 1 17 theme key 157:159 arg1 role 161:164 a key role 155:164 a key role 155:164 Human LOX-1/OLR 1 plays a key role in atherogenesis and endothelial dysfunction. 22688517 0 18 theme lipoprotein 99:109 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 18 theme lipoprotein 99:109 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 5 19 theme E 626:626 arg1 digestion 629:637 nonspecific protease (Pronase E) digestion 596:637 nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 596:701 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 8 20 theme new 1549:1551 arg1 light 1553:1557 new light 1549:1557 new light 1549:1557 The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. 22688517 5 21 theme site-specific 725:737 arg1 information 755:765 site-specific N-glycosylation information 725:765 site-specific N-glycosylation information of recombinant human LOX-1 725:792 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 0 22 theme human 60:64 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 22 theme human 60:64 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 4 23 theme human 501:505 arg1 LOX-1 507:511 recombinant human LOX-1 489:511 recombinant human LOX-1 489:511 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 6 24 gly glycosylation 1145:1157 arg2 site 1159:1162 one potential glycosylation site 1131:1162 one potential glycosylation site of recombinant human LOX-1 on Asn(139) 1131:1201 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 24 gly glycosylation 1145:1157 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 24 gly glycosylation 1145:1157 arg2 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 24 gly glycosylation 1145:1157 arg2 one 1131:1133 one 1131:1133 one 1131:1133 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 0 25 theme density 91:97 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 25 theme density 91:97 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 26 theme Site-specific 0:12 arg1 identification 30:43 Site-specific N-glycosylation identification 0:43 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 0:129 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 5 27 theme nonspecific 596:606 arg1 digestion 629:637 nonspecific protease (Pronase E) digestion 596:637 nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 596:701 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 28 theme diagnostic 1345:1354 arg1 ions 1356:1359 diagnostic ions 1345:1359 diagnostic ions 1345:1359 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 3 29 theme LOX-1 389:393 arg1 pattern 378:384 the N-glycosylation pattern 358:384 the N-glycosylation pattern of LOX-1 358:393 However, the N-glycosylation pattern of LOX-1 has not been described yet. 22688517 6 30 contain carry 1238:1242 arg1 site 1159:1162 one potential glycosylation site 1131:1162 one potential glycosylation site of recombinant human LOX-1 on Asn(139) 1131:1201 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 30 contain carry 1238:1242 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 30 contain carry 1238:1242 arg2 N-glycans 1271:1279 heterogeneous complex type N-glycans 1244:1279 heterogeneous complex type N-glycans 1244:1279 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 5 31 theme structure 822:830 arg1 confirmation 832:843 glycan structure confirmation 815:843 glycan structure confirmation 815:843 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 32 theme attached 978:985 arg1 LOX-1 995:999 LOX-1 995:999 LOX-1 995:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 32 theme attached 978:985 arg1 site 987:990 their corresponding attached site 958:990 N-glycans structures as well as their corresponding attached site of LOX-1 926:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 33 theme data 1330:1333 arg1 annotation 1302:1311 manual annotation 1295:1311 manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans 1295:1458 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 0 34 theme recombinant 48:58 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 34 theme recombinant 48:58 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 5 35 theme LOX-1 788:792 arg1 information 755:765 site-specific N-glycosylation information 725:765 site-specific N-glycosylation information of recombinant human LOX-1 725:792 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 36 theme MS 1044:1045 arg1 analysis 1047:1054 direct MS analysis 1037:1054 direct MS analysis of glycopeptides from non-specific protease digestion 1037:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 37 theme multistage 1316:1325 arg1 data 1330:1333 multistage MS data 1316:1333 multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans 1316:1458 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 8 38 theme relationships 1605:1617 arg1 research 1570:1577 further research 1562:1577 further research of the structure-function relationships of LOX-1 N-glycan 1562:1635 The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. 22688517 0 39 theme lectin-like 66:76 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 39 theme lectin-like 66:76 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 1 40 theme Human 131:135 arg1 LOX-1/OLR 137:145 Human LOX-1/OLR 1 131:147 Human LOX-1/OLR 1 131:147 Human LOX-1/OLR 1 plays a key role in atherogenesis and endothelial dysfunction. 22688517 8 41 theme LOX-1 1622:1626 arg1 N-glycan 1628:1635 LOX-1 N-glycan 1622:1635 LOX-1 N-glycan 1622:1635 The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. 22688517 6 42 theme glycosylation 1145:1157 arg1 site 1159:1162 one potential glycosylation site 1131:1162 one potential glycosylation site of recombinant human LOX-1 on Asn(139) 1131:1201 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 42 theme glycosylation 1145:1157 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 4 43 gly N-glycosylation 470:484 arg1 LOX-1 507:511 recombinant human LOX-1 489:511 recombinant human LOX-1 489:511 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 5 44 theme MS 665:666 arg1 analysis 694:701 MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 651:701 MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 651:701 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 45 theme multistage 672:681 arg1 MS 683:684 multistage MS 672:684 multistage MS 672:684 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 46 theme proper 1478:1483 arg1 interpretation 1493:1506 proper spectra interpretation 1478:1506 proper spectra interpretation 1478:1506 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 6 47 theme recombinant 1167:1177 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 1 48 theme endothelial 187:197 arg1 dysfunction 199:209 endothelial dysfunction 187:209 endothelial dysfunction 187:209 Human LOX-1/OLR 1 plays a key role in atherogenesis and endothelial dysfunction. 22688517 7 49 gly glycopeptides 1434:1446 arg2 glycopeptides 1434:1446 glycopeptides 1434:1446 glycopeptides 1434:1446 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 5 50 theme recombinant 770:780 arg1 LOX-1 788:792 recombinant human LOX-1 770:792 recombinant human LOX-1 770:792 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 51 theme tandem 891:896 arg1 MS. 898:900 tandem MS. 891:900 tandem MS. 891:900 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 52 theme MALDI-QIT-TOF 651:663 arg1 MS 687:688 MS(3) 687:691 MS(3) 687:691 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 52 theme MALDI-QIT-TOF 651:663 arg1 MS 665:666 MALDI-QIT-TOF MS 651:666 MALDI-QIT-TOF MS 651:666 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 6 53 theme human 1179:1183 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 54 theme heterogeneous 1244:1256 arg1 N-glycans 1271:1279 heterogeneous complex type N-glycans 1244:1279 heterogeneous complex type N-glycans 1244:1279 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 4 55 theme N-glycosylation 549:563 arg1 sites 565:569 N-glycosylation sites 549:569 N-glycosylation sites 549:569 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 5 56 theme N-glycans 926:934 arg1 structures 936:945 N-glycans structures 926:945 N-glycans structures as well as their corresponding attached site of LOX-1 926:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 2 57 gly N-glycosylation 216:230 arg1 LOX-1 235:239 LOX-1 235:239 LOX-1 235:239 The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. 22688517 2 58 theme biological 270:279 arg1 functions 281:289 its biological functions 266:289 its biological functions in vivo 266:297 The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. 22688517 6 59 theme LOX-1 1185:1189 arg1 site 1159:1162 one potential glycosylation site 1131:1162 one potential glycosylation site of recombinant human LOX-1 on Asn(139) 1131:1201 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 59 theme LOX-1 1185:1189 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 5 60 theme N-glycosylation 739:753 arg1 information 755:765 site-specific N-glycosylation information 725:765 site-specific N-glycosylation information of recombinant human LOX-1 725:792 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 61 theme protease 1091:1098 arg1 digestion 1100:1108 non-specific protease digestion 1078:1108 non-specific protease digestion 1078:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 0 62 theme receptor-1 111:120 arg1 identification 30:43 Site-specific N-glycosylation identification 0:43 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 0:129 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 4 63 theme N-glycan 528:535 arg1 profile 537:543 N-glycan profile 528:543 N-glycan profile 528:543 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 5 64 theme Pronase 618:624 arg1 digestion 629:637 nonspecific protease (Pronase E) digestion 596:637 nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 596:701 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 4 65 theme recombinant 489:499 arg1 LOX-1 507:511 recombinant human LOX-1 489:511 recombinant human LOX-1 489:511 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 5 66 from digestion 1100:1108 arg1 glycopeptides 1059:1071 glycopeptides 1059:1071 glycopeptides from non-specific protease digestion 1059:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 66 from digestion 1100:1108 arg1 analysis 1047:1054 direct MS analysis 1037:1054 direct MS analysis of glycopeptides from non-specific protease digestion 1037:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 67 theme LOX-1 995:999 arg1 structures 936:945 N-glycans structures 926:945 N-glycans structures as well as their corresponding attached site of LOX-1 926:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 67 theme LOX-1 995:999 arg1 LOX-1 995:999 LOX-1 995:999 LOX-1 995:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 67 theme LOX-1 995:999 arg1 site 987:990 their corresponding attached site 958:990 N-glycans structures as well as their corresponding attached site of LOX-1 926:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 3 68 theme N-glycosylation 362:376 arg1 pattern 378:384 the N-glycosylation pattern 358:384 the N-glycosylation pattern of LOX-1 358:393 However, the N-glycosylation pattern of LOX-1 has not been described yet. 22688517 4 69 theme LOX-1 507:511 arg1 N-glycosylation 470:484 the N-glycosylation 466:484 the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites 466:569 The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. 22688517 0 70 theme N-glycosylation 14:28 arg1 identification 30:43 Site-specific N-glycosylation identification 0:43 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 0:129 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 5 71 theme glycan 815:820 arg1 confirmation 832:843 glycan structure confirmation 815:843 glycan structure confirmation 815:843 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 72 theme protease 608:615 arg1 digestion 629:637 nonspecific protease (Pronase E) digestion 596:637 nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis 596:701 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 73 theme corresponding 964:976 arg1 LOX-1 995:999 LOX-1 995:999 LOX-1 995:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 5 73 theme corresponding 964:976 arg1 site 987:990 their corresponding attached site 958:990 N-glycans structures as well as their corresponding attached site of LOX-1 926:999 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 7 74 theme MS 1327:1328 arg1 data 1330:1333 multistage MS data 1316:1333 multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans 1316:1458 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 8 75 theme structure-function 1586:1603 arg1 relationships 1605:1617 the structure-function relationships 1582:1617 the structure-function relationships of LOX-1 N-glycan 1582:1635 The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. 22688517 5 76 theme human 782:786 arg1 LOX-1 788:792 recombinant human LOX-1 770:792 recombinant human LOX-1 770:792 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 3 77 gly N-glycosylation 362:376 arg1 LOX-1 389:393 LOX-1 389:393 LOX-1 389:393 However, the N-glycosylation pattern of LOX-1 has not been described yet. 22688517 5 78 theme direct 1037:1042 arg1 analysis 1047:1054 direct MS analysis 1037:1054 direct MS analysis of glycopeptides from non-specific protease digestion 1037:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 6 79 from site 1159:1162 arg1 139 1198:1200 139 1198:1200 139 1198:1200 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 79 from site 1159:1162 arg1 Asn 1194:1196 Asn 1194:1196 Asn(139) 1194:1201 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 0 80 theme oxidized 78:85 arg1 LOX-1 123:127 LOX-1 123:127 LOX-1 123:127 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 0 80 theme oxidized 78:85 arg1 receptor-1 111:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 48:120 recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) 48:128 Site-specific N-glycosylation identification of recombinant human lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). 22688517 8 81 theme further 1562:1568 arg1 research 1570:1577 further research 1562:1577 further research of the structure-function relationships of LOX-1 N-glycan 1562:1635 The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. 22688517 7 82 theme manual 1295:1300 arg1 annotation 1302:1311 manual annotation 1295:1311 manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans 1295:1458 In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. 22688517 8 83 theme N-glycan 1628:1635 arg1 relationships 1605:1617 the structure-function relationships 1582:1617 the structure-function relationships of LOX-1 N-glycan 1582:1635 The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. 22688517 5 84 dep using 885:889 arg1 reveal 914:919 reveal 914:919 reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion 914:1108 Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS(3)) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. 22688517 2 85 theme atherosclerosis 332:346 arg1 pathogenesis 316:327 the pathogenesis 312:327 the pathogenesis of atherosclerosis 312:346 The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. 22688517 6 86 theme potential 1135:1143 arg1 site 1159:1162 one potential glycosylation site 1131:1162 one potential glycosylation site of recombinant human LOX-1 on Asn(139) 1131:1201 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 22688517 6 86 theme potential 1135:1143 arg1 LOX-1 1185:1189 recombinant human LOX-1 1167:1189 recombinant human LOX-1 1167:1189 With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn(139) is readily identified and found to carry heterogeneous complex type N-glycans. 18420026 3 0 theme kDa 867:869 arg1 weights 836:842 molecular weights 826:842 molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14 826:1030 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 5 1 theme mouse 1286:1290 arg1 Necl1 1292:1296 mouse Necl1 1286:1296 mouse Necl1 1286:1296 There was no O-glycosylation site in either human NECL1 or mouse Necl1. 18420026 1 2 theme neural 224:229 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 2 theme neural 224:229 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 2 3 theme mouse 551:555 arg1 brains 557:562 mouse brains 551:562 mouse brains 551:562 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 7 4 theme increased 1656:1664 arg1 activity 1675:1682 an increased adhesion activity 1653:1682 an increased adhesion activity 1653:1682 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 7 5 dep showed 1636:1641 arg1 activity 1675:1682 an increased adhesion activity 1653:1682 an increased adhesion activity 1653:1682 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 6 6 with treatment 1339:1347 arg1 tissue 1372:1377 human fetal brain tissue 1354:1377 human fetal brain tissue 1354:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 7 theme transient 1396:1404 arg1 transfection 1406:1417 transient transfection 1396:1417 transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells 1396:1488 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 1 8 theme immunoglobulin-like 247:265 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 8 theme immunoglobulin-like 247:265 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 9 with glycoprotein 1539:1550 arg1 site 1580:1583 a single glycosylation site 1557:1583 a single glycosylation site at position N290KS 1557:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 10 theme kDa 671:673 arg1 weights 651:657 molecular weights 641:657 molecular weights of about 62 kDa and 48 kDa 641:684 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 1 11 theme bundles 437:443 arg1 formation 409:417 the formation 405:417 the formation of synapses, axon bundles and myelinated axons 405:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 2 12 theme developmental 577:589 arg1 stages 591:596 different developmental stages 567:596 different developmental stages 567:596 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 3 13 theme mouse 935:939 arg1 E16 941:943 mouse E16 935:943 mouse E16 935:943 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 5 14 gly O-glycosylation 1240:1254 arg2 site 1256:1259 no O-glycosylation site 1237:1259 no O-glycosylation site 1237:1259 There was no O-glycosylation site in either human NECL1 or mouse Necl1. 18420026 1 15 theme myelinated 449:458 arg1 axons 460:464 myelinated axons 449:464 myelinated axons 449:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 2 16 theme fetal 523:527 arg1 brains 539:544 human fetal and adult brains 517:544 human fetal and adult brains 517:544 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 0 17 theme adhesion 107:114 arg1 activity 116:123 its adhesion activity 103:123 its adhesion activity 103:123 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 1 18 contain has 301:303 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 18 contain has 301:303 arg2 activity 365:372 Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity 305:372 Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity 305:372 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 18 contain has 301:303 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 19 theme fetal 1360:1364 arg1 tissue 1372:1377 human fetal brain tissue 1354:1377 human fetal brain tissue 1354:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 20 gly glycoprotein 1539:1550 arg1 glycoprotein 1539:1550 an N-linked glycoprotein 1527:1550 an N-linked glycoprotein with a single glycosylation site at position N290KS 1527:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 20 gly glycoprotein 1539:1550 arg1 NECL1 1518:1522 human NECL1 1512:1522 human NECL1 1512:1522 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 21 theme N-linked 1530:1537 arg1 glycoprotein 1539:1550 an N-linked glycoprotein 1527:1550 an N-linked glycoprotein with a single glycosylation site at position N290KS 1527:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 21 theme N-linked 1530:1537 arg1 NECL1 1518:1522 human NECL1 1512:1522 human NECL1 1512:1522 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 22 theme fetal 706:710 arg1 brain 712:716 human fetal brain 700:716 human fetal brain 700:716 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 6 23 theme glycosylation 1443:1455 arg1 mutant 1462:1467 glycosylation site mutant 1443:1467 glycosylation site mutant 1443:1467 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 24 link N-linked 1530:1537 arg1 glycoprotein 1539:1550 an N-linked glycoprotein 1527:1550 an N-linked glycoprotein with a single glycosylation site at position N290KS 1527:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 24 link N-linked 1530:1537 arg1 NECL1 1518:1522 human NECL1 1512:1522 human NECL1 1512:1522 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 25 theme kDa 775:777 arg1 weight 756:761 a molecular weight 744:761 a molecular weight of about 48 kDa 744:777 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 7 26 gly glycosylation 1694:1706 arg2 site 1708:1711 the glycosylation site mutation 1690:1720 the glycosylation site mutation of NECL1 molecule 1690:1738 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 7 26 gly glycosylation 1694:1706 arg2 mutation 1713:1720 the glycosylation site mutation 1690:1720 the glycosylation site mutation of NECL1 molecule 1690:1738 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 2 27 theme adult 533:537 arg1 brains 539:544 human fetal and adult brains 517:544 human fetal and adult brains 517:544 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 3 28 with band 968:971 arg1 weight 990:995 a molecular weight 978:995 a molecular weight of about 48 kDa 978:1011 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 6 29 dep wild-type 1430:1438 arg1 NECL1 1469:1473 NECL1 1469:1473 NECL1 1469:1473 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 4 30 dep positions 1132:1140 arg1 positions 1132:1140 positions 1132:1140 positions N25LS and N290KS 1132:1157 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 30 dep positions 1132:1140 arg1 N290KS 1152:1157 N290KS 1152:1157 N290KS 1152:1157 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 30 dep positions 1132:1140 arg1 N25LS 1142:1146 N25LS 1142:1146 N25LS 1142:1146 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 2 31 from expression 494:503 arg1 brains 557:562 mouse brains 551:562 mouse brains 551:562 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 2 31 from expression 494:503 arg1 brains 539:544 human fetal and adult brains 517:544 human fetal and adult brains 517:544 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 0 32 theme single 48:53 arg1 site 71:74 a single N-glycosylation site 46:74 a single N-glycosylation site at N290KS 46:84 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 3 33 with band 734:737 arg1 weight 756:761 a molecular weight 744:761 a molecular weight of about 48 kDa 744:777 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 6 34 from transfection 1406:1417 arg1 results 1312:1318 the results 1308:1318 the results of N-Glycosidase F treatment with human fetal brain tissue 1308:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 34 from transfection 1406:1417 arg1 lysates 1383:1389 lysates 1383:1389 lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells 1383:1488 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 34 from transfection 1406:1417 arg1 cells 1484:1488 293ET cells 1478:1488 293ET cells 1478:1488 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 35 theme human 792:796 arg1 brain 804:808 human adult brain 792:808 human adult brain 792:808 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 1 36 theme homo- 324:328 arg1 activity 365:372 Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity 305:372 Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity 305:372 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 4 37 theme N-glycosylation 1088:1102 arg1 sites 1104:1108 two putative N-glycosylation sites 1075:1108 two putative N-glycosylation sites within human NECL1 1075:1127 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 6 38 with transfection 1406:1417 arg1 wild-type 1430:1438 wild-type 1430:1438 wild-type 1430:1438 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 38 with transfection 1406:1417 arg1 mutant 1462:1467 glycosylation site mutant 1443:1467 glycosylation site mutant 1443:1467 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 1 39 theme heterophilic 333:344 arg1 adhesion 356:363 heterophilic cell-cell adhesion 333:363 heterophilic cell-cell adhesion 333:363 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 0 40 gly N-glycosylation 55:69 arg2 N290KS 79:84 N290KS 79:84 N290KS 79:84 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 0 40 gly N-glycosylation 55:69 arg2 site 71:74 a single N-glycosylation site 46:74 a single N-glycosylation site at N290KS 46:84 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 1 41 theme Nectin-like 126:136 arg1 molecule 138:145 Nectin-like molecule 1 126:147 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 41 theme Nectin-like 126:136 arg1 NECL1 150:154 NECL1 150:154 NECL1 150:154 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 4 42 from positions 1185:1193 arg1 positions 1132:1140 positions 1132:1140 positions N25LS and N290KS 1132:1157 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 42 from positions 1185:1193 arg1 N290KS 1152:1157 N290KS 1152:1157 N290KS 1152:1157 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 42 from positions 1185:1193 arg1 N25LS 1142:1146 N25LS 1142:1146 N25LS 1142:1146 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 6 43 gly glycosylation 1443:1455 arg2 mutant 1462:1467 glycosylation site mutant 1443:1467 glycosylation site mutant 1443:1467 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 43 gly glycosylation 1443:1455 arg2 site 1457:1460 glycosylation site mutant 1443:1467 glycosylation site mutant 1443:1467 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 1 44 theme adhesion 356:363 arg1 activity 365:372 Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity 305:372 Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity 305:372 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 45 from now 190:192 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 45 from now 190:192 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 46 theme single 1559:1564 arg1 site 1580:1583 a single glycosylation site 1557:1583 a single glycosylation site at position N290KS 1557:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 47 theme kDa 682:684 arg1 weights 651:657 molecular weights 641:657 molecular weights of about 62 kDa and 48 kDa 641:684 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 0 48 theme Nectin-like 0:10 arg1 glycoprotein 28:39 a glycoprotein 26:39 a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity 26:123 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 0 48 theme Nectin-like 0:10 arg1 molecule 12:19 Nectin-like molecule 1 0:21 Nectin-like molecule 1 0:21 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 1 49 theme cell-cell 267:275 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 49 theme cell-cell 267:275 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 0 50 from N290KS 79:84 arg1 site 71:74 a single N-glycosylation site 46:74 a single N-glycosylation site at N290KS 46:84 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 3 51 theme kDa 856:858 arg1 weights 836:842 molecular weights 826:842 molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14 826:1030 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 5 52 theme O-glycosylation 1240:1254 arg1 site 1256:1259 no O-glycosylation site 1237:1259 no O-glycosylation site 1237:1259 There was no O-glycosylation site in either human NECL1 or mouse Necl1. 18420026 3 53 theme kDa 1009:1011 arg1 weight 990:995 a molecular weight 978:995 a molecular weight of about 48 kDa 978:1011 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 3 54 located found 783:787 arg1 brain 804:808 human adult brain 792:808 human adult brain 792:808 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 3 54 located found 783:787 arg2 band 734:737 only one band 725:737 only one band with a molecular weight of about 48 kDa 725:777 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 7 55 theme glycosylation 1694:1706 arg1 mutation 1713:1720 the glycosylation site mutation 1690:1720 the glycosylation site mutation of NECL1 molecule 1690:1738 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 3 56 theme molecular 641:649 arg1 weights 651:657 molecular weights 641:657 molecular weights of about 62 kDa and 48 kDa 641:684 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 6 57 theme treatment 1339:1347 arg1 results 1312:1318 the results 1308:1318 the results of N-Glycosidase F treatment with human fetal brain tissue 1308:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 57 theme treatment 1339:1347 arg1 lysates 1383:1389 lysates 1383:1389 lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells 1383:1488 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 4 58 gly N-glycosylation 1088:1102 arg2 sites 1104:1108 two putative N-glycosylation sites 1075:1108 two putative N-glycosylation sites within human NECL1 1075:1127 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 58 gly N-glycosylation 1088:1102 arg2 two 1075:1077 two 1075:1077 two 1075:1077 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 7 59 theme molecule 1731:1738 arg1 mutation 1713:1720 the glycosylation site mutation 1690:1720 the glycosylation site mutation of NECL1 molecule 1690:1738 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 0 60 gly glycoprotein 28:39 arg1 glycoprotein 28:39 a glycoprotein 26:39 a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity 26:123 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 0 60 gly glycoprotein 28:39 arg1 molecule 12:19 Nectin-like molecule 1 0:21 Nectin-like molecule 1 0:21 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 6 61 theme human 1512:1516 arg1 glycoprotein 1539:1550 an N-linked glycoprotein 1527:1550 an N-linked glycoprotein with a single glycosylation site at position N290KS 1527:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 61 theme human 1512:1516 arg1 NECL1 1518:1522 human NECL1 1512:1522 human NECL1 1512:1522 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 62 theme N-Glycosidase 1323:1335 arg1 F 1337:1337 N-Glycosidase F 1323:1337 N-Glycosidase F treatment with human fetal brain tissue 1323:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 7 63 theme adhesion 1666:1673 arg1 activity 1675:1682 an increased adhesion activity 1653:1682 an increased adhesion activity 1653:1682 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 3 64 dep found 691:695 arg1 bands 815:819 two bands 811:819 two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14 811:1030 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 1 65 theme synapses 422:429 arg1 formation 409:417 the formation 405:417 the formation of synapses, axon bundles and myelinated axons 405:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 66 theme tissue-specific 231:245 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 66 theme tissue-specific 231:245 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 4 67 theme mouse 1170:1174 arg1 Necl1 1176:1180 mouse Necl1 1170:1180 mouse Necl1 1170:1180 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 1 68 theme axon 432:435 arg1 bundles 437:443 axon bundles 432:443 axon bundles 432:443 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 3 69 with bands 630:634 arg1 weights 651:657 molecular weights 641:657 molecular weights of about 62 kDa and 48 kDa 641:684 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 5 70 theme human 1271:1275 arg1 NECL1 1277:1281 human NECL1 1271:1281 human NECL1 1271:1281 There was no O-glycosylation site in either human NECL1 or mouse Necl1. 18420026 4 71 dep positions 1185:1193 arg1 positions 1185:1193 positions 1185:1193 positions N23LS and N288KS 1185:1210 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 71 dep positions 1185:1193 arg1 N288KS 1205:1210 N288KS 1205:1210 N288KS 1205:1210 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 4 71 dep positions 1185:1193 arg1 N23LS 1195:1199 N23LS 1195:1199 N23LS 1195:1199 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 2 72 theme different 567:575 arg1 stages 591:596 different developmental stages 567:596 different developmental stages 567:596 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 7 73 theme aggregation 1610:1620 arg1 assay 1622:1626 Cell aggregation assay 1605:1626 Cell aggregation assay 1605:1626 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 3 74 theme human 700:704 arg1 brain 712:716 human fetal brain 700:716 human fetal brain 700:716 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 1 75 theme axons 460:464 arg1 formation 409:417 the formation 405:417 the formation of synapses, axon bundles and myelinated axons 405:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 76 theme brain 1366:1370 arg1 tissue 1372:1377 human fetal brain tissue 1354:1377 human fetal brain tissue 1354:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 2 77 theme human 517:521 arg1 brains 539:544 human fetal and adult brains 517:544 human fetal and adult brains 517:544 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 6 78 theme human 1354:1358 arg1 tissue 1372:1377 human fetal brain tissue 1354:1377 human fetal brain tissue 1354:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 2 79 theme NECL1 508:512 arg1 expression 494:503 the expression 490:503 the expression of NECL1 in human fetal and adult brains, and mouse brains 490:562 Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. 18420026 4 80 theme Bioinformatics 1033:1046 arg1 analysis 1048:1055 Bioinformatics analysis 1033:1055 Bioinformatics analysis 1033:1055 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 1 81 theme important 387:395 arg1 role 397:400 an important role 384:400 an important role 384:400 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 82 theme site 1457:1460 arg1 mutant 1462:1467 glycosylation site mutant 1443:1467 glycosylation site mutant 1443:1467 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 83 located found 691:695 arg1 brain 712:716 human fetal brain 700:716 human fetal brain 700:716 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 3 83 located found 691:695 arg2 bands 630:634 two bands 626:634 two bands with molecular weights of about 62 kDa and 48 kDa 626:684 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 7 84 theme Cell 1605:1608 arg1 assay 1622:1626 Cell aggregation assay 1605:1626 Cell aggregation assay 1605:1626 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 0 85 theme N-glycosylation 55:69 arg1 site 71:74 a single N-glycosylation site 46:74 a single N-glycosylation site at N290KS 46:84 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 18420026 3 86 with bands 815:819 arg1 weights 836:842 molecular weights 826:842 molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14 826:1030 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 1 87 theme molecule 138:145 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 87 theme molecule 138:145 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 88 from position 1588:1595 arg1 site 1580:1583 a single glycosylation site 1557:1583 a single glycosylation site at position N290KS 1557:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 1 89 theme -independent 311:322 arg1 homo- 324:328 Ca(2+)-independent homo- 305:328 Ca(2+)-independent homo- 305:328 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 3 90 theme adult 798:802 arg1 brain 804:808 human adult brain 792:808 human adult brain 792:808 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 4 91 theme human 1117:1121 arg1 NECL1 1123:1127 human NECL1 1117:1127 human NECL1 1117:1127 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 3 92 theme molecular 746:754 arg1 weight 756:761 a molecular weight 744:761 a molecular weight of about 48 kDa 744:777 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 1 93 theme cell-cell 346:354 arg1 adhesion 356:363 heterophilic cell-cell adhesion 333:363 heterophilic cell-cell adhesion 333:363 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 6 94 theme glycosylation 1566:1578 arg1 site 1580:1583 a single glycosylation site 1557:1583 a single glycosylation site at position N290KS 1557:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 95 theme 293ET 1478:1482 arg1 cells 1484:1488 293ET cells 1478:1488 293ET cells 1478:1488 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 4 96 theme putative 1079:1086 arg1 sites 1104:1108 two putative N-glycosylation sites 1075:1108 two putative N-glycosylation sites within human NECL1 1075:1127 Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. 18420026 1 97 theme adhesion 277:284 arg1 molecule 286:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule 222:293 a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons 222:464 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 1 97 theme adhesion 277:284 arg1 /CADM3/IGSF4B/TSLL1/SynCAM3 156:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3 126:182 Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. 18420026 3 98 theme molecular 980:988 arg1 weight 990:995 a molecular weight 978:995 a molecular weight of about 48 kDa 978:1011 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 3 99 theme molecular 826:834 arg1 weights 836:842 molecular weights 826:842 molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14 826:1030 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 7 100 theme NECL1 1725:1729 arg1 molecule 1731:1738 NECL1 molecule 1725:1738 NECL1 molecule 1725:1738 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 7 101 theme site 1708:1711 arg1 mutation 1713:1720 the glycosylation site mutation 1690:1720 the glycosylation site mutation of NECL1 molecule 1690:1738 Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. 18420026 6 102 theme F 1337:1337 arg1 treatment 1339:1347 N-Glycosidase F treatment 1323:1347 N-Glycosidase F treatment with human fetal brain tissue 1323:1377 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 3 103 theme expression 877:886 arg1 level 888:892 level 888:892 level 888:892 The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. 18420026 6 104 gly glycosylation 1566:1578 arg2 N290KS 1597:1602 N290KS 1597:1602 N290KS 1597:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 104 gly glycosylation 1566:1578 arg2 position 1588:1595 position 1588:1595 position N290KS 1588:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 6 104 gly glycosylation 1566:1578 arg2 site 1580:1583 a single glycosylation site 1557:1583 a single glycosylation site at position N290KS 1557:1602 Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. 18420026 0 105 with glycoprotein 28:39 arg1 site 71:74 a single N-glycosylation site 46:74 a single N-glycosylation site at N290KS 46:84 Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. 26536155 3 0 theme small 719:723 arg1 portion 733:739 a small peptide portion 717:739 a small peptide portion 717:739 The use of unspecific proteases such as Pronase can largely overcome this problem by generating glycopeptides with a small peptide portion. 26536155 5 1 theme porous 1282:1287 arg1 carbon-LC-ESI-QTOF-MS/MS 1301:1324 porous graphitized carbon-LC-ESI-QTOF-MS/MS 1282:1324 porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1282:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 1 2 gly O-glycopeptides 222:236 arg2 O-glycopeptides 222:236 O-glycopeptides 222:236 O-glycopeptides 222:236 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 5 3 with glycoproteins 1218:1230 arg1 carbon-LC-ESI-QTOF-MS/MS 1301:1324 porous graphitized carbon-LC-ESI-QTOF-MS/MS 1282:1324 porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1282:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 5 3 with glycoproteins 1218:1230 arg1 phase 1272:1276 integrated, sequential C18 reverse phase 1237:1276 integrated, sequential C18 reverse phase 1237:1276 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 1 4 theme glycosylation 326:338 arg1 site 340:343 one glycosylation site 322:343 one glycosylation site 322:343 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 6 5 gly glycoprotein 1442:1453 arg1 glycoprotein 1442:1453 glycoprotein standards 1442:1463 glycoprotein standards 1442:1463 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 7 6 theme single 1791:1796 arg1 run 1805:1807 a single sample run 1789:1807 a single sample run 1789:1807 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 2 7 theme site 581:584 arg1 identification 586:599 site identification 581:599 site identification 581:599 Trypsin is by far the most commonly used protease in glycoproteomic studies; however, it often results in long peptides that can harbor more than one glycan which may hamper site identification. 26536155 7 8 gly O-glycopeptide 1715:1728 arg2 O-glycopeptide 1715:1728 O-glycopeptide analysis 1715:1737 O-glycopeptide analysis 1715:1737 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 4 9 gly glycopeptides 762:774 arg2 glycopeptides 762:774 the resulting glycopeptides 748:774 the resulting glycopeptides 748:774 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 9 gly glycopeptides 762:774 arg2 useful 785:790 useful 785:790 useful 785:790 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 7 10 theme glycopeptide 1760:1771 arg1 coverage 1773:1780 a good glycopeptide coverage 1753:1780 a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample 1753:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 4 11 theme heterogeneous 961:973 arg1 properties 991:1000 the very heterogeneous physicochemical properties 952:1000 the very heterogeneous physicochemical properties of the glycopeptides 952:1021 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 3 12 theme unspecific 613:622 arg1 proteases 624:632 unspecific proteases 613:632 unspecific proteases such as Pronase 613:648 The use of unspecific proteases such as Pronase can largely overcome this problem by generating glycopeptides with a small peptide portion. 26536155 3 12 theme unspecific 613:622 arg1 Pronase 642:648 Pronase 642:648 Pronase 642:648 The use of unspecific proteases such as Pronase can largely overcome this problem by generating glycopeptides with a small peptide portion. 26536155 1 13 gly macroheterogeneity 350:367 arg1 glycoprotein 393:404 the glycoprotein 389:404 the glycoprotein 389:404 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 14 attach attached 310:317 arg1 site 340:343 one glycosylation site 322:343 one glycosylation site 322:343 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 14 attach attached 310:317 arg2 glycoforms 299:308 different glycoforms 289:308 different glycoforms attached to one glycosylation site 289:343 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 5 15 gly glycoproteins 1218:1230 arg1 glycoproteins 1218:1230 Pronase treated glycoproteins 1202:1230 Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1202:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 6 16 theme IgG3 1524:1527 arg1 glycosylation 1501:1513 the glycosylation 1497:1513 the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1497:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 7 17 gly glycoproteins 1676:1688 arg1 glycoproteins 1676:1688 glycoproteins 1676:1688 glycoproteins 1676:1688 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 0 18 theme Carbon-Liquid 82:94 arg1 Spectrometry 140:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry 57:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 1 19 theme glycoprotein 393:404 arg1 microheterogeneity 269:286 the microheterogeneity 265:286 the microheterogeneity (different glycoforms attached to one glycosylation site) 265:344 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 19 theme glycoprotein 393:404 arg1 macroheterogeneity 350:367 macroheterogeneity 350:367 macroheterogeneity (site occupancy) 350:384 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 19 theme glycoprotein 393:404 arg1 occupancy 375:383 site occupancy 370:383 site occupancy 370:383 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 7 20 theme small 1835:1839 arg1 sample 1852:1857 sample 1852:1857 sample 1852:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 7 20 theme small 1835:1839 arg1 amounts 1841:1847 only small amounts 1830:1847 only small amounts of sample 1830:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 4 21 theme mass 803:806 arg1 spectrometric 808:820 tandem mass spectrometric 796:820 tandem mass spectrometric investigation 796:834 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 5 22 theme collision-induced 1380:1396 arg1 dissociation 1398:1409 enhanced-energy collision-induced dissociation 1364:1409 enhanced-energy collision-induced dissociation 1364:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 0 23 theme Ionization 124:133 arg1 Spectrometry 140:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry 57:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 5 24 theme site-specific 1154:1166 arg1 N- 1168:1169 site-specific N- 1154:1169 site-specific N- 1154:1169 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 7 25 theme site-specific 1694:1706 arg1 N- 1708:1709 site-specific N- 1694:1709 site-specific N- 1694:1709 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 2 26 theme used 443:446 arg1 Trypsin 407:413 Trypsin 407:413 Trypsin 407:413 Trypsin is by far the most commonly used protease in glycoproteomic studies; however, it often results in long peptides that can harbor more than one glycan which may hamper site identification. 26536155 2 26 theme used 443:446 arg1 protease 448:455 the most commonly used protease 425:455 the most commonly used protease in glycoproteomic studies 425:481 Trypsin is by far the most commonly used protease in glycoproteomic studies; however, it often results in long peptides that can harbor more than one glycan which may hamper site identification. 26536155 0 27 theme Spectrometry 140:151 arg1 Approach 153:160 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 4 28 theme glycan 1096:1101 arg1 moiety 1103:1108 the glycan moiety 1092:1108 the glycan moiety 1092:1108 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 0 29 theme Site-Specific 0:12 arg1 N- 22:23 Site-Specific Protein N- 0:23 Site-Specific Protein N- 0:23 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 1 30 gly glycosylation 326:338 arg2 site 340:343 one glycosylation site 322:343 one glycosylation site 322:343 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 30 gly glycosylation 326:338 arg2 one 322:324 one 322:324 one 322:324 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 6 31 theme uncharacterized 1563:1577 arg1 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 31 theme uncharacterized 1563:1577 arg1 site 1593:1596 the hitherto uncharacterized glycosylation site Asn392 1550:1603 the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1550:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 5 32 theme reverse 1264:1270 arg1 phase 1272:1276 integrated, sequential C18 reverse phase 1237:1276 integrated, sequential C18 reverse phase 1237:1276 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 6 33 gly glycosylation 1579:1591 arg2 Asn392 1598:1603 Asn392 1598:1603 Asn392 1598:1603 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 33 gly glycosylation 1579:1591 arg1 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 33 gly glycosylation 1579:1591 arg2 site 1593:1596 the hitherto uncharacterized glycosylation site Asn392 1550:1603 the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1550:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 33 gly glycosylation 1579:1591 arg2 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 7 34 theme sample 1798:1803 arg1 run 1805:1807 a single sample run 1789:1807 a single sample run 1789:1807 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 2 35 theme long 513:516 arg1 peptides 518:525 long peptides 513:525 long peptides that can harbor more than one glycan which may hamper site identification 513:599 Trypsin is by far the most commonly used protease in glycoproteomic studies; however, it often results in long peptides that can harbor more than one glycan which may hamper site identification. 26536155 4 36 theme 1D-LC-ESI-MS/MS 868:882 arg1 approaches 884:893 conventional 1D-LC-ESI-MS/MS approaches 855:893 conventional 1D-LC-ESI-MS/MS approaches 855:893 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 5 37 theme glycoproteins 1218:1230 arg1 N- 1168:1169 site-specific N- 1154:1169 site-specific N- 1154:1169 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 5 37 theme glycoproteins 1218:1230 arg1 analysis 1190:1197 O-glycopeptide analysis 1175:1197 O-glycopeptide analysis 1175:1197 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 6 38 gly glycosylation 1501:1513 arg1 IgG3 1524:1527 human IgG3 1518:1527 human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1518:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 7 39 gly glycopeptide 1760:1771 arg2 glycopeptide 1760:1771 a good glycopeptide coverage 1753:1780 a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample 1753:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 0 40 theme C18-Porous 59:68 arg1 Spectrometry 140:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry 57:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 5 41 theme integrated 1237:1246 arg1 phase 1272:1276 integrated, sequential C18 reverse phase 1237:1276 integrated, sequential C18 reverse phase 1237:1276 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 0 42 dep N- 22:23 arg1 Analysis 45:52 Analysis 45:52 Analysis 45:52 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 1 43 theme different 289:297 arg1 glycoforms 299:308 different glycoforms 289:308 different glycoforms attached to one glycosylation site 289:343 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 7 44 theme analytical 1629:1638 arg1 tool 1640:1643 This analytical tool 1624:1643 This analytical tool 1624:1643 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 6 45 theme human 1518:1522 arg1 IgG3 1524:1527 human IgG3 1518:1527 human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1518:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 46 theme CH3 1612:1614 arg1 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 3 47 theme peptide 725:731 arg1 portion 733:739 a small peptide portion 717:739 a small peptide portion 717:739 The use of unspecific proteases such as Pronase can largely overcome this problem by generating glycopeptides with a small peptide portion. 26536155 5 48 theme graphitized 1289:1299 arg1 carbon-LC-ESI-QTOF-MS/MS 1301:1324 porous graphitized carbon-LC-ESI-QTOF-MS/MS 1282:1324 porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1282:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 1 49 theme N- 215:216 arg1 analysis 203:210 The analysis 199:210 The analysis of N- and O-glycopeptides 199:236 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 50 gly glycoprotein 393:404 arg1 glycoprotein 393:404 the glycoprotein 389:404 the glycoprotein 389:404 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 51 theme O-glycopeptides 222:236 arg1 analysis 203:210 The analysis 199:210 The analysis of N- and O-glycopeptides 199:236 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 5 52 theme Pronase 1202:1208 arg1 glycoproteins 1218:1230 Pronase treated glycoproteins 1202:1230 Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1202:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 4 53 theme incomplete 907:916 arg1 glycosylation 918:930 incomplete glycosylation 907:930 incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety 907:1108 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 0 54 theme Pronase 168:174 arg1 Glycopeptides 184:196 Pronase Treated Glycopeptides 168:196 Pronase Treated Glycopeptides 168:196 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 5 55 gly O-glycopeptide 1175:1188 arg2 O-glycopeptide 1175:1188 O-glycopeptide analysis 1175:1197 O-glycopeptide analysis 1175:1197 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 5 56 theme universal 1131:1139 arg1 workflow 1141:1148 a universal workflow 1129:1148 a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1129:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 3 57 theme proteases 624:632 arg1 use 606:608 The use 602:608 The use of unspecific proteases such as Pronase 602:648 The use of unspecific proteases such as Pronase can largely overcome this problem by generating glycopeptides with a small peptide portion. 26536155 7 58 theme good 1755:1758 arg1 coverage 1773:1780 a good glycopeptide coverage 1753:1780 a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample 1753:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 4 59 theme glycopeptides 1009:1021 arg1 properties 991:1000 the very heterogeneous physicochemical properties 952:1000 the very heterogeneous physicochemical properties of the glycopeptides 952:1021 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 1 60 dep microheterogeneity 269:286 arg1 glycoforms 299:308 different glycoforms 289:308 different glycoforms attached to one glycosylation site 289:343 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 6 61 theme glycoprotein 1442:1453 arg1 standards 1455:1463 glycoprotein standards 1442:1463 glycoprotein standards 1442:1463 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 62 from details 1539:1545 arg1 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 62 from details 1539:1545 arg1 site 1593:1596 the hitherto uncharacterized glycosylation site Asn392 1550:1603 the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1550:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 4 63 gly glycopeptides 1009:1021 arg2 glycopeptides 1009:1021 the glycopeptides 1005:1021 the glycopeptides 1005:1021 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 64 theme physicochemical 975:989 arg1 properties 991:1000 the very heterogeneous physicochemical properties 952:1000 the very heterogeneous physicochemical properties of the glycopeptides 952:1021 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 65 theme resulting 752:760 arg1 useful 785:790 useful 785:790 useful 785:790 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 65 theme resulting 752:760 arg1 glycopeptides 762:774 the resulting glycopeptides 748:774 the resulting glycopeptides 748:774 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 0 66 theme Graphitized 70:80 arg1 Spectrometry 140:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry 57:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 7 67 theme O-glycopeptide 1715:1728 arg1 analysis 1730:1737 O-glycopeptide analysis 1715:1737 O-glycopeptide analysis 1715:1737 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 5 68 theme lower- 1353:1358 arg1 combination 1338:1348 a combination 1336:1348 a combination of lower- 1336:1358 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 5 68 theme lower- 1353:1358 arg1 dissociation 1398:1409 enhanced-energy collision-induced dissociation 1364:1409 enhanced-energy collision-induced dissociation 1364:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 0 69 theme Chromatography-Electrospray 96:122 arg1 Spectrometry 140:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry 57:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 7 70 theme sample 1852:1857 arg1 sample 1852:1857 sample 1852:1857 sample 1852:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 7 70 theme sample 1852:1857 arg1 amounts 1841:1847 only small amounts 1830:1847 only small amounts of sample 1830:1857 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 5 71 theme enhanced-energy 1364:1378 arg1 dissociation 1398:1409 enhanced-energy collision-induced dissociation 1364:1409 enhanced-energy collision-induced dissociation 1364:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 4 72 with analysis 841:848 arg1 approaches 884:893 conventional 1D-LC-ESI-MS/MS approaches 855:893 conventional 1D-LC-ESI-MS/MS approaches 855:893 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 0 73 theme Mass 135:138 arg1 Spectrometry 140:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry 57:151 a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach 57:160 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 5 74 theme treated 1210:1216 arg1 glycoproteins 1218:1230 Pronase treated glycoproteins 1202:1230 Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation 1202:1409 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 1 75 gly microheterogeneity 269:286 arg1 glycoprotein 393:404 the glycoprotein 389:404 the glycoprotein 389:404 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 5 76 dep integrated 1237:1246 arg1 sequential 1249:1258 sequential 1249:1258 sequential 1249:1258 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 4 77 theme tandem 796:801 arg1 spectrometric 808:820 tandem mass spectrometric 796:820 tandem mass spectrometric investigation 796:834 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 78 theme peptide 1040:1046 arg1 sequence 1048:1055 the peptide sequence 1036:1055 the peptide sequence 1036:1055 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 79 theme spectrometric 808:820 arg1 investigation 822:834 tandem mass spectrometric investigation 796:834 tandem mass spectrometric investigation 796:834 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 0 80 theme Protein 14:20 arg1 N- 22:23 Site-Specific Protein N- 0:23 Site-Specific Protein N- 0:23 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 5 81 theme O-glycopeptide 1175:1188 arg1 analysis 1190:1197 O-glycopeptide analysis 1175:1197 O-glycopeptide analysis 1175:1197 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 6 82 theme glycosylation 1579:1591 arg1 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 82 theme glycosylation 1579:1591 arg1 site 1593:1596 the hitherto uncharacterized glycosylation site Asn392 1550:1603 the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1550:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 5 83 theme C18 1260:1262 arg1 phase 1272:1276 integrated, sequential C18 reverse phase 1237:1276 integrated, sequential C18 reverse phase 1237:1276 Here, we describe a universal workflow for site-specific N- and O-glycopeptide analysis of Pronase treated glycoproteins with integrated, sequential C18 reverse phase and porous graphitized carbon-LC-ESI-QTOF-MS/MS employing a combination of lower- and enhanced-energy collision-induced dissociation. 26536155 4 84 theme moiety 1103:1108 arg1 charges 1081:1087 charges 1081:1087 charges 1081:1087 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 84 theme moiety 1103:1108 arg1 size 1072:1075 size 1072:1075 size 1072:1075 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 4 84 theme moiety 1103:1108 arg1 sequence 1048:1055 the peptide sequence 1036:1055 the peptide sequence 1036:1055 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 2 85 theme glycoproteomic 460:473 arg1 studies 475:481 glycoproteomic studies 460:481 glycoproteomic studies 460:481 Trypsin is by far the most commonly used protease in glycoproteomic studies; however, it often results in long peptides that can harbor more than one glycan which may hamper site identification. 26536155 7 86 theme glycoproteins 1676:1688 arg1 variety 1665:1671 a variety 1663:1671 a variety of glycoproteins 1663:1688 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 7 86 theme glycoproteins 1676:1688 arg1 glycoproteins 1676:1688 glycoproteins 1676:1688 glycoproteins 1676:1688 This analytical tool can be applied to a variety of glycoproteins for site-specific N- and O-glycopeptide analysis, resulting in a good glycopeptide coverage within a single sample run and, thus, requiring only small amounts of sample. 26536155 4 87 theme conventional 855:866 arg1 approaches 884:893 conventional 1D-LC-ESI-MS/MS approaches 855:893 conventional 1D-LC-ESI-MS/MS approaches 855:893 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 1 88 theme site 370:373 arg1 macroheterogeneity 350:367 macroheterogeneity 350:367 macroheterogeneity (site occupancy) 350:384 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 1 88 theme site 370:373 arg1 occupancy 375:383 site occupancy 370:383 site occupancy 370:383 The analysis of N- and O-glycopeptides remains challenging due to the microheterogeneity (different glycoforms attached to one glycosylation site) and macroheterogeneity (site occupancy) of the glycoprotein. 26536155 0 89 theme Treated 176:182 arg1 Glycopeptides 184:196 Pronase Treated Glycopeptides 168:196 Pronase Treated Glycopeptides 168:196 Site-Specific Protein N- and O-Glycosylation Analysis by a C18-Porous Graphitized Carbon-Liquid Chromatography-Electrospray Ionization Mass Spectrometry Approach Using Pronase Treated Glycopeptides. 26536155 2 90 from protease 448:455 arg1 studies 475:481 glycoproteomic studies 460:481 glycoproteomic studies 460:481 Trypsin is by far the most commonly used protease in glycoproteomic studies; however, it often results in long peptides that can harbor more than one glycan which may hamper site identification. 26536155 3 91 gly glycopeptides 698:710 arg2 glycopeptides 698:710 glycopeptides 698:710 glycopeptides 698:710 The use of unspecific proteases such as Pronase can largely overcome this problem by generating glycopeptides with a small peptide portion. 26536155 4 92 dep glycosylation 918:930 arg1 coverage 932:939 coverage 932:939 coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety 932:1108 While the resulting glycopeptides are very useful for tandem mass spectrometric investigation, the analysis with conventional 1D-LC-ESI-MS/MS approaches can lead to incomplete glycosylation coverage because of the very heterogeneous physicochemical properties of the glycopeptides depending on the peptide sequence as well as the size and charges of the glycan moiety. 26536155 6 93 theme domain 1616:1621 arg1 domain 1616:1621 the CH3 domain 1608:1621 the CH3 domain 1608:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 26536155 6 93 theme domain 1616:1621 arg1 site 1593:1596 the hitherto uncharacterized glycosylation site Asn392 1550:1603 the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain 1550:1621 The approach was evaluated on glycoprotein standards and also applied to investigate the glycosylation of human IgG3 providing details on the hitherto uncharacterized glycosylation site Asn392 of the CH3 domain. 28060820 0 0 theme Decisive 60:67 arg1 Role 69:72 a Decisive Role 58:72 a Decisive Role of N-Glycosylation 58:91 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 5 1 theme glycans 1051:1057 arg1 devoid 1041:1046 devoid 1041:1046 devoid 1041:1046 Proteolysis of an IL-6R completely devoid of glycans is significantly impaired. 28060820 5 2 theme devoid 1041:1046 arg1 IL-6R 1024:1028 an IL-6R 1021:1028 an IL-6R completely devoid of glycans 1021:1057 Proteolysis of an IL-6R completely devoid of glycans is significantly impaired. 28060820 4 3 from exosite 984:990 arg1 D1 1002:1003 domain D1 995:1003 domain D1 995:1003 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 4 4 theme N-glycan 975:982 arg1 exosite 984:990 an N-glycan exosite 972:990 an N-glycan exosite in domain D1 972:1003 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 1 5 theme proinflammatory 203:217 arg1 properties 219:228 the proinflammatory properties 199:228 the proinflammatory properties of IL-6 199:236 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 2 6 theme human 436:440 arg1 serum 442:446 human serum 436:446 human serum 436:446 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 4 7 theme domain 995:1000 arg1 D1 1002:1003 domain D1 995:1003 domain D1 995:1003 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 2 8 theme sites 600:604 arg1 occupancy 560:568 the occupancy 556:568 the occupancy of all O- and N-glycosylation sites of the human sIL-6R 556:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 6 9 theme important 1112:1120 arg1 regulator 1122:1130 an important regulator 1109:1130 an important regulator for sIL-6R generation 1109:1152 Thus, glycosylation is an important regulator for sIL-6R generation. 28060820 6 9 theme important 1112:1120 arg1 glycosylation 1092:1104 glycosylation 1092:1104 glycosylation 1092:1104 Thus, glycosylation is an important regulator for sIL-6R generation. 28060820 3 10 theme Val-356 745:751 arg1 mutation 733:740 mutation 733:740 mutation of Val-356 733:751 The metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) uses this cleavage site in vitro, and mutation of Val-356 is sufficient to completely abrogate IL-6R proteolysis. 28060820 2 11 theme N-glycosylation 584:598 arg1 sites 600:604 all O- and N-glycosylation sites 573:604 sites 600:604 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 11 theme N-glycosylation 584:598 arg1 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 12 gly N-glycosylation 584:598 arg2 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 12 gly N-glycosylation 584:598 arg1 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 12 gly N-glycosylation 584:598 arg2 sites 600:604 all O- and N-glycosylation sites 573:604 sites 600:604 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 13 theme proteolytic 469:479 arg1 cleavage 481:488 proteolytic cleavage 469:488 proteolytic cleavage 469:488 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 5 14 theme IL-6R 1024:1028 arg1 Proteolysis 1006:1016 Proteolysis 1006:1016 Proteolysis of an IL-6R completely devoid of glycans 1006:1057 Proteolysis of an IL-6R completely devoid of glycans is significantly impaired. 28060820 2 15 theme O- 577:578 arg1 sites 600:604 all O- and N-glycosylation sites 573:604 sites 600:604 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 15 theme O- 577:578 arg1 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 16 gly occupancy 560:568 arg2 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 16 gly occupancy 560:568 arg2 sites 600:604 all O- and N-glycosylation sites 573:604 sites 600:604 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 0 17 theme N-Glycosylation 77:91 arg1 Origin 12:17 Proteolytic Origin 0:17 Proteolytic Origin of the Soluble Human IL-6R In Vivo 0:52 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 0 17 theme N-Glycosylation 77:91 arg1 Role 69:72 a Decisive Role 58:72 a Decisive Role of N-Glycosylation 58:91 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 4 18 theme cleavage 954:961 arg1 site 963:966 the cleavage site 950:966 the cleavage site 950:966 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 2 19 from form 428:431 arg1 serum 442:446 human serum 436:446 human serum 436:446 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 20 theme sIL-6R 619:624 arg1 sites 600:604 all O- and N-glycosylation sites 573:604 sites 600:604 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 20 theme sIL-6R 619:624 arg1 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 1 21 theme soluble 149:155 arg1 sIL-6R 172:177 sIL-6R 172:177 sIL-6R 172:177 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 1 21 theme soluble 149:155 arg1 receptor 162:169 its soluble IL-6 receptor 145:169 its soluble IL-6 receptor (sIL-6R) 145:178 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 0 22 theme Proteolytic 0:10 arg1 Origin 12:17 Proteolytic Origin 0:17 Proteolytic Origin of the Soluble Human IL-6R In Vivo 0:52 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 2 23 theme human 613:617 arg1 sIL-6R 619:624 the human sIL-6R 609:624 the human sIL-6R 609:624 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 1 24 theme IL-6 157:160 arg1 sIL-6R 172:177 sIL-6R 172:177 sIL-6R 172:177 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 1 24 theme IL-6 157:160 arg1 receptor 162:169 its soluble IL-6 receptor 145:169 its soluble IL-6 receptor (sIL-6R) 145:178 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 3 25 dep metalloprotease 631:645 arg1 metalloproteinase 665:681 metalloproteinase 17 665:684 metalloproteinase 17 (ADAM17) 665:693 The metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) uses this cleavage site in vitro, and mutation of Val-356 is sufficient to completely abrogate IL-6R proteolysis. 28060820 3 25 dep metalloprotease 631:645 arg1 ADAM17 687:692 ADAM17 687:692 ADAM17 687:692 The metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) uses this cleavage site in vitro, and mutation of Val-356 is sufficient to completely abrogate IL-6R proteolysis. 28060820 3 25 dep metalloprotease 631:645 arg1 disintegrin 649:659 a disintegrin 647:659 a disintegrin 647:659 The metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) uses this cleavage site in vitro, and mutation of Val-356 is sufficient to completely abrogate IL-6R proteolysis. 28060820 2 26 theme liquid 366:371 arg1 spectrometry 393:404 liquid chromatography-mass spectrometry 366:404 liquid chromatography-mass spectrometry 366:404 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 2 27 theme chromatography-mass 373:391 arg1 spectrometry 393:404 liquid chromatography-mass spectrometry 366:404 liquid chromatography-mass spectrometry 366:404 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 4 28 from site 963:966 arg1 D1 1002:1003 domain D1 995:1003 domain D1 995:1003 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 2 29 theme cleavage 499:506 arg1 site 508:511 its cleavage site 495:511 its cleavage site between Pro-355 and Val-356 495:539 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 0 30 theme Human 34:38 arg1 IL-6R 40:44 the Soluble Human IL-6R 22:44 the Soluble Human IL-6R In Vivo 22:52 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 1 31 theme attractive 257:266 arg1 target 280:285 an attractive therapeutic target 254:285 an attractive therapeutic target 254:285 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 0 32 theme Soluble 26:32 arg1 IL-6R 40:44 the Soluble Human IL-6R 22:44 the Soluble Human IL-6R In Vivo 22:52 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 1 33 theme therapeutic 268:278 arg1 target 280:285 an attractive therapeutic target 254:285 an attractive therapeutic target 254:285 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 4 34 theme IL-6R 870:874 arg1 signaling 853:861 signaling 853:861 signaling of the IL-6R 853:874 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 2 35 theme sIL-6R 421:426 arg1 form 428:431 an sIL-6R form 418:431 an sIL-6R form in human serum that originates from proteolytic cleavage 418:488 Here, we use liquid chromatography-mass spectrometry to identify an sIL-6R form in human serum that originates from proteolytic cleavage, map its cleavage site between Pro-355 and Val-356, and determine the occupancy of all O- and N-glycosylation sites of the human sIL-6R. 28060820 0 36 theme IL-6R 40:44 arg1 Origin 12:17 Proteolytic Origin 0:17 Proteolytic Origin of the Soluble Human IL-6R In Vivo 0:52 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 0 36 theme IL-6R 40:44 arg1 Role 69:72 a Decisive Role 58:72 a Decisive Role of N-Glycosylation 58:91 Proteolytic Origin of the Soluble Human IL-6R In Vivo and a Decisive Role of N-Glycosylation. 28060820 3 37 theme cleavage 705:712 arg1 site 714:717 this cleavage site 700:717 this cleavage site 700:717 The metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) uses this cleavage site in vitro, and mutation of Val-356 is sufficient to completely abrogate IL-6R proteolysis. 28060820 4 38 dep N- 916:917 arg1 sequon 938:943 sequon 938:943 sequon near the cleavage site and an N-glycan exosite in domain D1 938:1003 N- and O-glycosylation were dispensable for signaling of the IL-6R, but proteolysis was orchestrated by an N- and O-glycosylated sequon near the cleavage site and an N-glycan exosite in domain D1. 28060820 3 39 theme IL-6R 790:794 arg1 proteolysis 796:806 IL-6R proteolysis 790:806 IL-6R proteolysis 790:806 The metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) uses this cleavage site in vitro, and mutation of Val-356 is sufficient to completely abrogate IL-6R proteolysis. 28060820 6 40 theme sIL-6R 1136:1141 arg1 generation 1143:1152 sIL-6R generation 1136:1152 sIL-6R generation 1136:1152 Thus, glycosylation is an important regulator for sIL-6R generation. 28060820 1 41 theme cytokine 111:118 arg1 IL-6 135:138 IL-6 135:138 IL-6 135:138 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 1 41 theme cytokine 111:118 arg1 interleukin-6 120:132 the cytokine interleukin-6 107:132 the cytokine interleukin-6 (IL-6) 107:139 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 1 42 theme IL-6 233:236 arg1 properties 219:228 the proinflammatory properties 199:228 the proinflammatory properties of IL-6 199:236 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28060820 1 43 theme interleukin-6 120:132 arg1 Signaling 94:102 Signaling 94:102 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) 94:178 Signaling of the cytokine interleukin-6 (IL-6) via its soluble IL-6 receptor (sIL-6R) is responsible for the proinflammatory properties of IL-6 and constitutes an attractive therapeutic target, but how the sIL-6R is generated in vivo remains largely unclear. 28668641 0 0 theme serpin 60:65 arg1 activity 67:74 serpin activity 60:74 serpin activity 60:74 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 1 1 theme conformational 304:317 arg1 changes 319:325 conformational changes 304:325 conformational changes underlying serpin function 304:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 2 2 from effects 433:439 arg1 properties 456:465 biochemical properties 444:465 biochemical properties relevant to vaspin function 444:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 1 3 with glycoprotein 128:139 arg1 sites 176:180 three predicted glycosylation sites 146:180 three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 146:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 2 4 theme relevant 467:474 arg1 properties 456:465 biochemical properties 444:465 biochemical properties relevant to vaspin function 444:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 4 5 theme binding 672:678 arg1 affinity 680:687 binding affinity 672:687 binding affinity to heparin 672:698 Although binding affinity to heparin is slightly decreased, the protease inhibition reaction is still significantly accelerated in the presence of heparin. 28668641 1 6 theme reactive 233:240 arg1 loop 249:252 the reactive center loop 229:252 the reactive center loop 229:252 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 0 7 from Glycosylation 0:12 arg1 stability 105:113 thermal stability 97:113 thermal stability 97:113 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 7 from Glycosylation 0:12 arg1 activity 67:74 serpin activity 60:74 serpin activity 60:74 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 7 from Glycosylation 0:12 arg1 binding 85:91 heparin binding 77:91 heparin binding 77:91 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 1 8 gly glycosylation 162:174 arg2 residues 196:203 asparagine residues 185:203 asparagine residues 185:203 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 8 gly glycosylation 162:174 arg2 sites 176:180 three predicted glycosylation sites 146:180 three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 146:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 8 gly glycosylation 162:174 arg2 three 146:150 three 146:150 three 146:150 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 9 theme center 242:247 arg1 loop 249:252 the reactive center loop 229:252 the reactive center loop 229:252 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 10 theme serpin 338:343 arg1 function 345:352 serpin function 338:352 serpin function 338:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 11 from residues 196:203 arg1 sites 176:180 three predicted glycosylation sites 146:180 three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 146:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 4 12 theme inhibition 736:745 arg1 reaction 747:754 the protease inhibition reaction 723:754 the protease inhibition reaction 723:754 Although binding affinity to heparin is slightly decreased, the protease inhibition reaction is still significantly accelerated in the presence of heparin. 28668641 5 13 theme thermal 848:854 arg1 stability 856:864 thermal stability 848:864 thermal stability 848:864 Glycosylation did not affect thermal stability. 28668641 0 14 theme heparin 77:83 arg1 binding 85:91 heparin binding 77:91 heparin binding 77:91 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 1 15 theme predicted 152:160 arg1 sites 176:180 three predicted glycosylation sites 146:180 three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 146:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 2 16 theme vaspin 418:423 arg1 effects 433:439 its effects 429:439 its effects on biochemical properties relevant to vaspin function 429:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 2 16 theme vaspin 418:423 arg1 glycosylation 395:407 the glycosylation 391:407 the glycosylation of human vaspin 391:423 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 1 17 theme glycosylation 162:174 arg1 sites 176:180 three predicted glycosylation sites 146:180 three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 146:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 0 18 theme vaspin 23:28 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of human vaspin (SERPINA12) 0:40 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 18 theme vaspin 23:28 arg1 impact 50:55 its impact 46:55 its impact on serpin activity, heparin binding and thermal stability 46:113 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 19 theme thermal 97:103 arg1 stability 105:113 thermal stability 97:113 thermal stability 97:113 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 4 20 theme heparin 810:816 arg1 presence 798:805 the presence 794:805 the presence of heparin 794:816 Although binding affinity to heparin is slightly decreased, the protease inhibition reaction is still significantly accelerated in the presence of heparin. 28668641 4 21 theme protease 727:734 arg1 reaction 747:754 the protease inhibition reaction 723:754 the protease inhibition reaction 723:754 Although binding affinity to heparin is slightly decreased, the protease inhibition reaction is still significantly accelerated in the presence of heparin. 28668641 0 22 theme human 17:21 arg1 SERPINA12 31:39 SERPINA12 31:39 SERPINA12 31:39 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 22 theme human 17:21 arg1 vaspin 23:28 human vaspin 17:28 human vaspin (SERPINA12) 17:40 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 2 23 gly glycosylation 395:407 arg1 vaspin 418:423 human vaspin 412:423 human vaspin 412:423 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 2 23 gly glycosylation 395:407 arg1 properties 456:465 biochemical properties 444:465 biochemical properties relevant to vaspin function 444:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 2 23 gly glycosylation 395:407 arg1 study 363:367 this study 358:367 this study 358:367 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 2 24 theme biochemical 444:454 arg1 properties 456:465 biochemical properties 444:465 biochemical properties relevant to vaspin function 444:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 0 25 gly Glycosylation 0:12 arg1 SERPINA12 31:39 SERPINA12 31:39 SERPINA12 31:39 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 25 gly Glycosylation 0:12 arg1 vaspin 23:28 human vaspin 17:28 human vaspin (SERPINA12) 17:40 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 1 26 theme asparagine 185:194 arg1 residues 196:203 asparagine residues 185:203 asparagine residues 185:203 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 0 27 from impact 50:55 arg1 stability 105:113 thermal stability 97:113 thermal stability 97:113 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 27 from impact 50:55 arg1 activity 67:74 serpin activity 60:74 serpin activity 60:74 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 0 27 from impact 50:55 arg1 binding 85:91 heparin binding 77:91 heparin binding 77:91 Glycosylation of human vaspin (SERPINA12) and its impact on serpin activity, heparin binding and thermal stability. 28668641 2 28 theme human 412:416 arg1 vaspin 418:423 human vaspin 412:423 human vaspin 412:423 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 1 29 from proximity 216:224 arg1 located 205:211 located 205:211 located 205:211 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 30 from located 205:211 arg1 proximity 216:224 proximity 216:224 proximity to the reactive center loop 216:252 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 31 theme important 285:293 arg1 roles 295:299 important roles 285:299 important roles 285:299 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 3 32 theme kallikrein 649:658 arg1 inhibition 615:624 inhibition 615:624 inhibition of the target protease kallikrein 7 615:660 We show that vaspin is modified at all three sites and biochemical data demonstrate that glycosylation does not hinder inhibition of the target protease kallikrein 7. 28668641 3 33 theme biochemical 551:561 arg1 data 563:566 biochemical data 551:566 biochemical data 551:566 We show that vaspin is modified at all three sites and biochemical data demonstrate that glycosylation does not hinder inhibition of the target protease kallikrein 7. 28668641 1 34 gly glycoprotein 128:139 arg1 Vaspin 116:121 Vaspin 116:121 Vaspin 116:121 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 34 gly glycoprotein 128:139 arg1 glycoprotein 128:139 a glycoprotein 126:139 a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 126:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 1 35 theme located 205:211 arg1 sites 176:180 three predicted glycosylation sites 146:180 three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function 146:352 Vaspin is a glycoprotein with three predicted glycosylation sites at asparagine residues located in proximity to the reactive center loop and close to domains that play important roles in conformational changes underlying serpin function. 28668641 3 36 theme target 633:638 arg1 kallikrein 649:658 the target protease kallikrein 7 629:660 the target protease kallikrein 7 629:660 We show that vaspin is modified at all three sites and biochemical data demonstrate that glycosylation does not hinder inhibition of the target protease kallikrein 7. 28668641 2 37 theme vaspin 479:484 arg1 function 486:493 vaspin function 479:493 vaspin function 479:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28668641 3 38 theme protease 640:647 arg1 kallikrein 649:658 the target protease kallikrein 7 629:660 the target protease kallikrein 7 629:660 We show that vaspin is modified at all three sites and biochemical data demonstrate that glycosylation does not hinder inhibition of the target protease kallikrein 7. 28668641 2 39 from glycosylation 395:407 arg1 properties 456:465 biochemical properties 444:465 biochemical properties relevant to vaspin function 444:493 In this study, we have investigated the glycosylation of human vaspin and its effects on biochemical properties relevant to vaspin function. 28082515 10 0 theme immunofluorescence 1505:1522 arg1 studies 1524:1530 our interactomic and immunofluorescence studies 1484:1530 studies 1524:1530 Intriguingly, our interactomic and immunofluorescence studies showed that ΔITILELP is partially retained at the endoplasmic reticulum (ER). 28082515 3 1 theme cystinosin 508:517 arg1 activity 519:526 residual cystinosin activity 499:526 residual cystinosin activity as a transporter 499:543 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 1 2 theme cystine 260:266 arg1 accumulation 244:255 intralysosomal accumulation 229:255 intralysosomal accumulation of cystine 229:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 12 3 theme low 1964:1966 arg1 activity 1978:1985 low transport activity 1964:1985 low transport activity 1964:1985 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 7 4 theme wild-type 1077:1085 arg1 cystinosin 1087:1096 wild-type cystinosin 1077:1096 wild-type cystinosin 1077:1096 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 11 5 theme glycosylation 1834:1846 arg1 sites 1848:1852 its remaining glycosylation sites 1820:1852 its remaining glycosylation sites 1820:1852 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 8 6 theme ΔITILELP 1222:1229 arg1 instability 1231:1241 ΔITILELP instability 1222:1241 ΔITILELP instability 1222:1241 Additional lysosome inhibition experiments confirmed ΔITILELP instability and showed that the degradation was mainly lysosomal. 28082515 11 7 theme ΔITILELP 1631:1638 arg1 mutation 1640:1647 the ΔITILELP mutation 1627:1647 the ΔITILELP mutation 1627:1647 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 2 8 theme causative 273:281 arg1 CTNS 306:309 CTNS 306:309 CTNS 306:309 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 2 8 theme causative 273:281 arg1 gene 283:286 The causative gene 269:286 The causative gene for cystinosis 269:301 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 12 9 theme state 1938:1942 arg1 turnover 1878:1885 the high turnover 1869:1885 the high turnover of ΔITILELP 1869:1897 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 12 9 theme state 1938:1942 arg1 responsible 1997:2007 responsible 1997:2007 responsible 1997:2007 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 10 10 theme endoplasmic 1582:1592 arg1 ER 1605:1606 ER 1605:1606 ER 1605:1606 Intriguingly, our interactomic and immunofluorescence studies showed that ΔITILELP is partially retained at the endoplasmic reticulum (ER). 28082515 10 10 theme endoplasmic 1582:1592 arg1 reticulum 1594:1602 the endoplasmic reticulum 1578:1602 the endoplasmic reticulum (ER) 1578:1607 Intriguingly, our interactomic and immunofluorescence studies showed that ΔITILELP is partially retained at the endoplasmic reticulum (ER). 28082515 3 11 theme protein-protein 573:587 arg1 interactions 589:600 its protein-protein interactions 569:600 its protein-protein interactions 569:600 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 9 12 theme pathway 1427:1433 arg1 complex 1390:1396 the V-ATPase complex 1377:1396 the V-ATPase complex 1377:1396 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 9 12 theme pathway 1427:1433 arg1 members 1407:1413 some members 1402:1413 some members of the mTOR pathway, similar to the wild-type protein 1402:1467 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 11 13 theme sites 1848:1852 arg1 sites 1848:1852 its remaining glycosylation sites 1820:1852 its remaining glycosylation sites 1820:1852 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 11 13 theme sites 1848:1852 arg1 six 1813:1815 six 1813:1815 six 1813:1815 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 9 14 theme similar 1436:1442 arg1 pathway 1427:1433 the mTOR pathway 1418:1433 the mTOR pathway 1418:1433 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 8 15 theme lysosome 1180:1187 arg1 experiments 1200:1210 Additional lysosome inhibition experiments 1169:1210 Additional lysosome inhibition experiments 1169:1210 Additional lysosome inhibition experiments confirmed ΔITILELP instability and showed that the degradation was mainly lysosomal. 28082515 7 16 theme culture 1035:1041 arg1 study 1051:1055 cell culture (SILAC) study 1030:1055 cell culture (SILAC) study 1030:1055 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 5 17 theme consensus 812:820 arg1 site 822:825 a consensus site 810:825 a consensus site on N66, one of the protein's seven glycosylation sites 810:880 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 7 18 theme isotope 995:1001 arg1 labeling 1003:1010 Our dynamic stable isotope labeling 976:1010 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study 976:1055 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 12 19 theme immature 1915:1922 arg1 state 1938:1942 its immature glycosylation state 1911:1942 its immature glycosylation state in combination with low transport activity 1911:1985 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 5 20 gly glycosylation 862:874 arg2 seven 856:860 seven 856:860 seven 856:860 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 5 20 gly glycosylation 862:874 arg2 sites 876:880 the protein's seven glycosylation sites 842:880 the protein's seven glycosylation sites 842:880 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 11 21 gly glycosylation 1834:1846 arg2 sites 1848:1852 its remaining glycosylation sites 1820:1852 its remaining glycosylation sites 1820:1852 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 4 22 theme severe 704:709 arg1 forms 711:715 severe forms 704:715 severe forms 704:715 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 11 23 theme high-mannose 1785:1796 arg1 glycans 1798:1804 high-mannose glycans 1785:1804 high-mannose glycans 1785:1804 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 7 24 theme dynamic 980:986 arg1 labeling 1003:1010 Our dynamic stable isotope labeling 976:1010 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study 976:1055 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 7 25 theme amino 1015:1019 arg1 acids 1021:1025 amino acids 1015:1025 amino acids 1015:1025 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 0 26 theme Differential 59:70 arg1 Labeling 95:102 Differential Dynamic Stable Isotope Labeling 59:102 Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) 59:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 12 27 with combination 1947:1957 arg1 activity 1978:1985 low transport activity 1964:1985 low transport activity 1964:1985 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 5 28 theme glycosylation 862:874 arg1 sites 876:880 the protein's seven glycosylation sites 842:880 the protein's seven glycosylation sites 842:880 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 0 29 theme Stable 80:85 arg1 Labeling 95:102 Differential Dynamic Stable Isotope Labeling 59:102 Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) 59:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 0 30 from Impact 0:5 arg1 Stability 46:54 Protein Stability 38:54 Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) 38:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 12 31 theme ΔITILELP 1890:1897 arg1 turnover 1878:1885 the high turnover 1869:1885 the high turnover of ΔITILELP 1869:1897 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 12 31 theme ΔITILELP 1890:1897 arg1 responsible 1997:2007 responsible 1997:2007 responsible 1997:2007 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 6 32 theme faster 956:961 arg1 degradation 963:973 faster degradation 956:973 faster degradation 956:973 Our hypothesis was that the ΔITILELP mutant is less stable and undergoes faster degradation. 28082515 0 33 theme Amino 107:111 arg1 Acids 113:117 Amino Acids 107:117 Amino Acids in Cell Culture (SILAC) 107:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 9 34 from lysosome 1321:1328 arg1 capable 1349:1355 capable 1349:1355 capable 1349:1355 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 4 35 theme residual 740:747 arg1 activity 761:768 its residual transported activity 736:768 its residual transported activity 736:768 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 0 36 theme Cystinosin 10:19 arg1 Glycosylation 21:33 Cystinosin Glycosylation 10:33 Cystinosin Glycosylation 10:33 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 3 37 theme varying 447:453 arg1 severity 463:470 varying disease severity 447:470 varying disease severity 447:470 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 12 38 from state 1938:1942 arg1 combination 1947:1957 combination 1947:1957 combination with low transport activity 1947:1985 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 7 39 dep rapidly 1160:1166 arg1 times 1149:1153 three times 1143:1153 three times 1143:1153 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 12 40 located observed 2027:2034 arg1 patients 2044:2051 some patients 2039:2051 some patients 2039:2051 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 12 40 located observed 2027:2034 arg2 phenotype 2017:2025 the phenotype 2013:2025 the phenotype observed in some patients 2013:2051 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 2 41 theme proton-driven 362:374 arg1 cystinosin 338:347 the protein cystinosin 326:347 the protein cystinosin 326:347 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 2 41 theme proton-driven 362:374 arg1 transporter 384:394 a lysosomal proton-driven cystine transporter 350:394 a lysosomal proton-driven cystine transporter 350:394 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 1 42 theme intralysosomal 229:242 arg1 accumulation 244:255 intralysosomal accumulation 229:255 intralysosomal accumulation of cystine 229:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 11 43 contain carries 1777:1783 arg1 ΔITILELP 1768:1775 ΔITILELP 1768:1775 ΔITILELP 1768:1775 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 11 43 contain carries 1777:1783 arg3 six 1813:1815 six 1813:1815 six 1813:1815 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 11 43 contain carries 1777:1783 arg2 glycans 1798:1804 high-mannose glycans 1785:1804 high-mannose glycans 1785:1804 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 11 43 contain carries 1777:1783 arg3 sites 1848:1852 its remaining glycosylation sites 1820:1852 its remaining glycosylation sites 1820:1852 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 3 44 theme residual 499:506 arg1 activity 519:526 residual cystinosin activity 499:526 residual cystinosin activity as a transporter 499:543 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 11 45 theme protein 1656:1662 arg1 misfolding 1664:1673 protein misfolding 1656:1673 protein misfolding 1656:1673 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 1 46 theme rare 160:163 arg1 Cystinosis 144:153 Cystinosis 144:153 Cystinosis 144:153 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 1 46 theme rare 160:163 arg1 disorder 203:210 a rare autosomal recessive lysosomal storage disorder 158:210 a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine 158:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 12 47 theme transport 1968:1976 arg1 activity 1978:1985 low transport activity 1964:1985 low transport activity 1964:1985 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 3 48 with correlation 482:492 arg1 activity 519:526 residual cystinosin activity 499:526 residual cystinosin activity as a transporter 499:543 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 3 48 with correlation 482:492 arg1 maintenance 554:564 maintenance 554:564 maintenance of its protein-protein interactions 554:600 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 9 49 theme mTOR 1422:1425 arg1 pathway 1427:1433 the mTOR pathway 1418:1433 the mTOR pathway 1418:1433 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 12 50 theme glycosylation 1924:1936 arg1 state 1938:1942 its immature glycosylation state 1911:1942 its immature glycosylation state in combination with low transport activity 1911:1985 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 4 51 theme only 657:660 arg1 mutation 643:650 the ΔITILELP mutation 630:650 the ΔITILELP mutation 630:650 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 4 51 theme only 657:660 arg1 mutation 662:669 the only mutation 653:669 the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity 653:768 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 0 52 theme Cell 122:125 arg1 Culture 127:133 Cell Culture 122:133 Cell Culture (SILAC) 122:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 0 52 theme Cell 122:125 arg1 SILAC 136:140 SILAC 136:140 SILAC 136:140 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 1 53 theme recessive 175:183 arg1 Cystinosis 144:153 Cystinosis 144:153 Cystinosis 144:153 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 1 53 theme recessive 175:183 arg1 disorder 203:210 a rare autosomal recessive lysosomal storage disorder 158:210 a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine 158:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 2 54 theme protein 330:336 arg1 CTNS 306:309 CTNS 306:309 CTNS 306:309 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 2 54 theme protein 330:336 arg1 cystinosin 338:347 the protein cystinosin 326:347 the protein cystinosin 326:347 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 2 54 theme protein 330:336 arg1 transporter 384:394 a lysosomal proton-driven cystine transporter 350:394 a lysosomal proton-driven cystine transporter 350:394 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 7 55 theme cell 1030:1033 arg1 culture 1035:1041 cell culture 1030:1041 cell culture (SILAC) study 1030:1055 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 7 55 theme cell 1030:1033 arg1 SILAC 1044:1048 SILAC 1044:1048 SILAC 1044:1048 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 1 56 theme autosomal 165:173 arg1 Cystinosis 144:153 Cystinosis 144:153 Cystinosis 144:153 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 1 56 theme autosomal 165:173 arg1 disorder 203:210 a rare autosomal recessive lysosomal storage disorder 158:210 a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine 158:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 1 57 theme storage 195:201 arg1 Cystinosis 144:153 Cystinosis 144:153 Cystinosis 144:153 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 1 57 theme storage 195:201 arg1 disorder 203:210 a rare autosomal recessive lysosomal storage disorder 158:210 a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine 158:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 8 58 theme Additional 1169:1178 arg1 experiments 1200:1210 Additional lysosome inhibition experiments 1169:1210 Additional lysosome inhibition experiments 1169:1210 Additional lysosome inhibition experiments confirmed ΔITILELP instability and showed that the degradation was mainly lysosomal. 28082515 4 59 theme ΔITILELP 634:641 arg1 mutation 643:650 the ΔITILELP mutation 630:650 the ΔITILELP mutation 630:650 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 4 59 theme ΔITILELP 634:641 arg1 mutation 662:669 the only mutation 653:669 the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity 653:768 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 3 60 theme interactions 589:600 arg1 maintenance 554:564 maintenance 554:564 maintenance of its protein-protein interactions 554:600 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 9 61 theme wild-type 1451:1459 arg1 protein 1461:1467 the wild-type protein 1447:1467 the wild-type protein 1447:1467 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 8 62 theme inhibition 1189:1198 arg1 experiments 1200:1210 Additional lysosome inhibition experiments 1169:1210 Additional lysosome inhibition experiments 1169:1210 Additional lysosome inhibition experiments confirmed ΔITILELP instability and showed that the degradation was mainly lysosomal. 28082515 6 63 theme ΔITILELP 911:918 arg1 mutant 920:925 the ΔITILELP mutant 907:925 the ΔITILELP mutant 907:925 Our hypothesis was that the ΔITILELP mutant is less stable and undergoes faster degradation. 28082515 6 63 theme ΔITILELP 911:918 arg1 stable 935:940 stable 935:940 stable 935:940 Our hypothesis was that the ΔITILELP mutant is less stable and undergoes faster degradation. 28082515 0 64 from Acids 113:117 arg1 Culture 127:133 Cell Culture 122:133 Cell Culture (SILAC) 122:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 0 64 from Acids 113:117 arg1 SILAC 136:140 SILAC 136:140 SILAC 136:140 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 4 65 with inconsistent 718:729 arg1 activity 761:768 its residual transported activity 736:768 its residual transported activity 736:768 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 7 66 theme stable 988:993 arg1 labeling 1003:1010 Our dynamic stable isotope labeling 976:1010 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study 976:1055 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 11 67 theme Golgi 1726:1730 arg1 apparatus 1732:1740 the Golgi apparatus 1722:1740 the Golgi apparatus 1722:1740 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 0 68 theme Dynamic 72:78 arg1 Labeling 95:102 Differential Dynamic Stable Isotope Labeling 59:102 Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) 59:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 0 69 theme Isotope 87:93 arg1 Labeling 95:102 Differential Dynamic Stable Isotope Labeling 59:102 Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) 59:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 9 70 from capable 1349:1355 arg1 lysosome 1321:1328 the lysosome 1317:1328 the lysosome 1317:1328 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 28082515 5 71 theme sites 876:880 arg1 sites 876:880 the protein's seven glycosylation sites 842:880 the protein's seven glycosylation sites 842:880 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 5 71 theme sites 876:880 arg1 one 835:837 one 835:837 one 835:837 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 5 71 theme sites 876:880 arg1 N66 830:832 N66 830:832 N66 830:832 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 4 72 theme transported 749:759 arg1 activity 761:768 its residual transported activity 736:768 its residual transported activity 736:768 In this study, we focus on the ΔITILELP mutation, the only mutation reported that sometimes leads to severe forms, inconsistent with its residual transported activity. 28082515 5 73 from site 822:825 arg1 sites 876:880 the protein's seven glycosylation sites 842:880 the protein's seven glycosylation sites 842:880 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 5 73 from site 822:825 arg1 one 835:837 one 835:837 one 835:837 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 5 73 from site 822:825 arg1 N66 830:832 N66 830:832 N66 830:832 ΔITILELP is a deletion that eliminates a consensus site on N66, one of the protein's seven glycosylation sites. 28082515 1 74 theme lysosomal 185:193 arg1 Cystinosis 144:153 Cystinosis 144:153 Cystinosis 144:153 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 1 74 theme lysosomal 185:193 arg1 disorder 203:210 a rare autosomal recessive lysosomal storage disorder 158:210 a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine 158:266 Cystinosis is a rare autosomal recessive lysosomal storage disorder characterized by intralysosomal accumulation of cystine. 28082515 7 75 dep stable 1106:1111 arg1 whereas 1114:1120 whereas 1114:1120 whereas 1114:1120 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 12 76 theme high 1873:1876 arg1 turnover 1878:1885 the high turnover 1869:1885 the high turnover of ΔITILELP 1869:1897 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 12 76 theme high 1873:1876 arg1 responsible 1997:2007 responsible 1997:2007 responsible 1997:2007 We found that the high turnover of ΔITILELP, because of its immature glycosylation state in combination with low transport activity, might be responsible for the phenotype observed in some patients. 28082515 2 77 theme cystine 376:382 arg1 cystinosin 338:347 the protein cystinosin 326:347 the protein cystinosin 326:347 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 2 77 theme cystine 376:382 arg1 transporter 384:394 a lysosomal proton-driven cystine transporter 350:394 a lysosomal proton-driven cystine transporter 350:394 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 0 78 theme Glycosylation 21:33 arg1 Impact 0:5 Impact 0:5 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 0:142 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 11 79 theme ER 1676:1677 arg1 retention 1679:1687 ER retention 1676:1687 ER retention 1676:1687 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 0 80 theme Protein 38:44 arg1 Stability 46:54 Protein Stability 38:54 Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) 38:141 Impact of Cystinosin Glycosylation on Protein Stability by Differential Dynamic Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). 28082515 3 81 theme disease 455:461 arg1 severity 463:470 varying disease severity 447:470 varying disease severity 447:470 Over 100 mutations have been reported, leading to varying disease severity, often in correlation with residual cystinosin activity as a transporter and with maintenance of its protein-protein interactions. 28082515 11 82 theme remaining 1824:1832 arg1 sites 1848:1852 its remaining glycosylation sites 1820:1852 its remaining glycosylation sites 1820:1852 We proposed that the ΔITILELP mutation causes protein misfolding, ER retention and inability to be processed in the Golgi apparatus, and we demonstrated that ΔITILELP carries high-mannose glycans on all six of its remaining glycosylation sites. 28082515 7 83 from labeling 1003:1010 arg1 study 1051:1055 cell culture (SILAC) study 1030:1055 cell culture (SILAC) study 1030:1055 Our dynamic stable isotope labeling by amino acids in cell culture (SILAC) study clearly showed that wild-type cystinosin is very stable, whereas ΔITILELP is degraded three times more rapidly. 28082515 2 84 theme lysosomal 352:360 arg1 cystinosin 338:347 the protein cystinosin 326:347 the protein cystinosin 326:347 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 2 84 theme lysosomal 352:360 arg1 transporter 384:394 a lysosomal proton-driven cystine transporter 350:394 a lysosomal proton-driven cystine transporter 350:394 The causative gene for cystinosis is CTNS, which encodes the protein cystinosin, a lysosomal proton-driven cystine transporter. 28082515 9 85 theme V-ATPase 1381:1388 arg1 complex 1390:1396 the V-ATPase complex 1377:1396 the V-ATPase complex 1377:1396 We observed that in the lysosome, ΔITILELP is still capable of interacting with the V-ATPase complex and some members of the mTOR pathway, similar to the wild-type protein. 21515415 5 0 theme N-glycosylation 711:725 arg1 site 727:730 each N-glycosylation site 706:730 each N-glycosylation site 706:730 The size of the carbohydrate at each N-glycosylation site ranged from 3.14 to 4.2kDa. 21515415 7 1 theme PLTP 1065:1068 arg1 activity 1070:1077 PLTP activity 1065:1077 PLTP activity 1065:1077 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 2 dep resulted 1020:1027 arg1 compared 1140:1147 compared 1140:1147 compared to wild type 1140:1160 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 8 3 theme PLTP 1369:1372 arg1 activity 1404:1411 its phospholipid transfer activity 1378:1411 its phospholipid transfer activity 1378:1411 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 3 theme PLTP 1369:1372 arg1 secretion 1356:1364 cellular secretion 1347:1364 cellular secretion of PLTP 1347:1372 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 1 4 theme transfer 163:170 arg1 PLTP 181:184 PLTP 181:184 PLTP 181:184 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 1 4 theme transfer 163:170 arg1 protein 172:178 The plasma phospholipid transfer protein 139:178 The plasma phospholipid transfer protein (PLTP) 139:185 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 5 5 theme carbohydrate 690:701 arg1 size 678:681 The size 674:681 The size of the carbohydrate at each N-glycosylation site 674:730 The size of the carbohydrate at each N-glycosylation site ranged from 3.14 to 4.2kDa. 21515415 0 6 theme specific 76:83 arg1 activity 85:92 specific activity 76:92 specific activity 76:92 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 8 7 theme cellular 1347:1354 arg1 secretion 1356:1364 cellular secretion 1347:1364 cellular secretion of PLTP 1347:1372 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 7 8 theme significant 1086:1096 arg1 decrease 1105:1112 a significant 29-80% decrease 1084:1112 a significant 29-80% decrease in PLTP specific activity 1084:1138 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 0 9 from Impact 0:5 arg1 activity 63:70 activity 63:70 activity 63:70 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 0 9 from Impact 0:5 arg1 activity 85:92 specific activity 76:92 specific activity 76:92 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 0 9 from Impact 0:5 arg1 secretion 52:60 cellular secretion 43:60 cellular secretion 43:60 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 7 10 theme wild 1152:1155 arg1 type 1157:1160 wild type 1152:1160 wild type 1152:1160 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 2 11 contain has 244:246 arg1 It 241:242 It 241:242 It 241:242 It has six potential N-glycosylation sites. 21515415 2 11 contain has 244:246 arg2 sites 278:282 six potential N-glycosylation sites 248:282 six potential N-glycosylation sites 248:282 It has six potential N-glycosylation sites. 21515415 5 12 gly N-glycosylation 711:725 arg2 site 727:730 each N-glycosylation site 706:730 each N-glycosylation site 706:730 The size of the carbohydrate at each N-glycosylation site ranged from 3.14 to 4.2kDa. 21515415 6 13 dep secretion 922:930 arg1 % 934:934 8% 933:934 8% 933:934 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 6 13 dep secretion 922:930 arg1 % 939:939 14% 937:939 14% 937:939 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 6 13 dep secretion 922:930 arg1 % 947:947 32% 945:947 32% 945:947 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 8 14 from impact 1337:1342 arg1 activity 1404:1411 its phospholipid transfer activity 1378:1411 its phospholipid transfer activity 1378:1411 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 14 from impact 1337:1342 arg1 secretion 1356:1364 cellular secretion 1347:1364 cellular secretion of PLTP 1347:1372 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 7 15 theme glycosylation 1000:1012 arg1 sites 1014:1018 the six glycosylation sites 992:1018 the six glycosylation sites 992:1018 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 3 16 theme Flp-In 492:497 arg1 cells 499:503 Chinese hamster ovary Flp-In cells 470:503 Chinese hamster ovary Flp-In cells 470:503 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 0 17 theme phospholipid 108:119 arg1 protein 130:136 the plasma phospholipid transfer protein 97:136 the plasma phospholipid transfer protein 97:136 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 6 18 theme modest 844:849 arg1 enhancement 851:861 a modest enhancement 842:861 a modest enhancement (15% and 60%) 842:875 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 4 19 used utilized 664:671 arg2 sites 638:642 all six sites 630:642 all six sites 630:642 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 2 20 theme N-glycosylation 262:276 arg1 sites 278:282 six potential N-glycosylation sites 248:282 six potential N-glycosylation sites 248:282 It has six potential N-glycosylation sites. 21515415 7 21 attach Removal 951:957 arg2 N-glycosylation 962:976 N-glycosylation 962:976 N-glycosylation 962:976 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 21 attach Removal 951:957 arg3 sites 1014:1018 the six glycosylation sites 992:1018 the six glycosylation sites 992:1018 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 21 attach Removal 951:957 arg3 one 985:987 one 985:987 one 985:987 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 0 22 theme plasma 101:106 arg1 protein 130:136 the plasma phospholipid transfer protein 97:136 the plasma phospholipid transfer protein 97:136 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 3 23 theme alanine 386:392 arg1 mutations 400:408 alanine point mutations 386:408 alanine point mutations 386:408 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 7 24 from decrease 1105:1112 arg1 activity 1070:1077 PLTP activity 1065:1077 PLTP activity 1065:1077 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 24 from decrease 1105:1112 arg1 activity 1131:1138 PLTP specific activity 1117:1138 PLTP specific activity 1117:1138 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 2 25 theme potential 252:260 arg1 sites 278:282 six potential N-glycosylation sites 248:282 six potential N-glycosylation sites 248:282 It has six potential N-glycosylation sites. 21515415 8 26 theme carbohydrate 1298:1309 arg1 chains 1311:1316 the carbohydrate chains 1294:1316 the carbohydrate chains 1294:1316 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 3 27 theme PLTP 323:326 arg1 secretion 328:336 PLTP secretion 323:336 PLTP secretion 323:336 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 8 28 theme transfer 1395:1402 arg1 activity 1404:1411 its phospholipid transfer activity 1378:1411 its phospholipid transfer activity 1378:1411 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 6 29 from reduction 909:917 arg1 secretion 922:930 secretion 922:930 secretion (8%, 14% and 32%) 922:948 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 8 30 theme N-linked 1207:1214 arg1 requirement 1240:1250 a requirement 1238:1250 a requirement for secretion or activity 1238:1276 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 30 theme N-linked 1207:1214 arg1 chain 1229:1233 no single N-linked carbohydrate chain 1197:1233 no single N-linked carbohydrate chain 1197:1233 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 6 31 from effect 764:769 arg1 secretion 820:828 PLTP secretion 815:828 PLTP secretion 815:828 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 7 32 theme significant 1034:1044 arg1 decrease 1053:1060 a significant 35-78% decrease 1032:1060 a significant 35-78% decrease in PLTP activity 1032:1077 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 8 33 theme phospholipid 1382:1393 arg1 activity 1404:1411 its phospholipid transfer activity 1378:1411 its phospholipid transfer activity 1378:1411 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 3 34 contain containing 365:374 arg1 variants 356:363 six variants 352:363 six variants containing serine to alanine point mutations 352:408 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 3 34 contain containing 365:374 arg2 serine 376:381 serine 376:381 serine to alanine point mutations 376:408 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 8 35 theme chains 1311:1316 arg1 removal 1283:1289 the removal 1279:1289 the removal of the carbohydrate chains 1279:1316 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 0 36 theme N-glycosylation 24:38 arg1 Impact 0:5 Impact 0:5 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 0:137 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 0 37 theme protein 130:136 arg1 activity 63:70 activity 63:70 activity 63:70 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 0 37 theme protein 130:136 arg1 activity 85:92 specific activity 76:92 specific activity 76:92 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 0 37 theme protein 130:136 arg1 secretion 52:60 cellular secretion 43:60 cellular secretion 43:60 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 7 38 from one 985:987 arg1 Removal 951:957 Removal 951:957 Removal of N-glycosylation at any one of the six glycosylation sites 951:1018 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 39 theme N-glycosylation 962:976 arg1 Removal 951:957 Removal 951:957 Removal of N-glycosylation at any one of the six glycosylation sites 951:1018 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 0 40 theme site-specific 10:22 arg1 N-glycosylation 24:38 site-specific N-glycosylation 10:38 site-specific N-glycosylation 10:38 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 4 41 theme PLTP 543:546 arg1 mutants 548:554 the six PLTP mutants 535:554 the six PLTP mutants 535:554 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 0 42 theme transfer 121:128 arg1 protein 130:136 the plasma phospholipid transfer protein 97:136 the plasma phospholipid transfer protein 97:136 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 8 43 theme carbohydrate 1216:1227 arg1 requirement 1240:1250 a requirement 1238:1250 a requirement for secretion or activity 1238:1276 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 43 theme carbohydrate 1216:1227 arg1 chain 1229:1233 no single N-linked carbohydrate chain 1197:1233 no single N-linked carbohydrate chain 1197:1233 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 1 44 theme key 195:197 arg1 role 199:202 a key role 193:202 a key role 193:202 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 0 45 theme cellular 43:50 arg1 secretion 52:60 cellular secretion 43:60 cellular secretion 43:60 Impact of site-specific N-glycosylation on cellular secretion, activity and specific activity of the plasma phospholipid transfer protein. 21515415 7 46 theme specific 1122:1129 arg1 activity 1131:1138 PLTP specific activity 1117:1138 PLTP specific activity 1117:1138 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 47 theme 29-80 1098:1102 arg1 % 1103:1103 % 1103:1103 % 1103:1103 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 3 48 theme site-directed 427:439 arg1 mutagenesis 441:451 site-directed mutagenesis 427:451 site-directed mutagenesis 427:451 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 3 49 theme point 394:398 arg1 mutations 400:408 alanine point mutations 386:408 alanine point mutations 386:408 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 7 50 from decrease 1053:1060 arg1 activity 1070:1077 PLTP activity 1065:1077 PLTP activity 1065:1077 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 50 from decrease 1053:1060 arg1 activity 1131:1138 PLTP specific activity 1117:1138 PLTP specific activity 1117:1138 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 51 theme sites 1014:1018 arg1 sites 1014:1018 the six glycosylation sites 992:1018 the six glycosylation sites 992:1018 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 51 theme sites 1014:1018 arg1 one 985:987 one 985:987 one 985:987 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 6 52 theme PLTP 815:818 arg1 secretion 820:828 PLTP secretion 815:828 PLTP secretion 815:828 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 4 53 theme wild 587:590 arg1 type 592:595 wild type 587:595 wild type 587:595 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 2 54 gly N-glycosylation 262:276 arg2 six 248:250 six 248:250 six 248:250 It has six potential N-glycosylation sites. 21515415 2 54 gly N-glycosylation 262:276 arg2 sites 278:282 six potential N-glycosylation sites 248:282 six potential N-glycosylation sites 248:282 It has six potential N-glycosylation sites. 21515415 4 55 theme apparent 510:517 arg1 less 569:572 less 569:572 less 569:572 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 4 55 theme apparent 510:517 arg1 size 519:522 The apparent size 506:522 The apparent size of each of the six PLTP mutants 506:554 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 8 56 link N-linked 1207:1214 arg1 requirement 1240:1250 a requirement 1238:1250 a requirement for secretion or activity 1238:1276 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 56 link N-linked 1207:1214 arg1 chain 1229:1233 no single N-linked carbohydrate chain 1197:1233 no single N-linked carbohydrate chain 1197:1233 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 57 theme single 1200:1205 arg1 requirement 1240:1250 a requirement 1238:1250 a requirement for secretion or activity 1238:1276 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 57 theme single 1200:1205 arg1 chain 1229:1233 no single N-linked carbohydrate chain 1197:1233 no single N-linked carbohydrate chain 1197:1233 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 3 58 theme hamster 478:484 arg1 cells 499:503 Chinese hamster ovary Flp-In cells 470:503 Chinese hamster ovary Flp-In cells 470:503 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 4 59 theme each 527:530 arg1 less 569:572 less 569:572 less 569:572 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 4 59 theme each 527:530 arg1 size 519:522 The apparent size 506:522 The apparent size of each of the six PLTP mutants 506:554 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 7 60 theme % 1051:1051 arg1 decrease 1053:1060 a significant 35-78% decrease 1032:1060 a significant 35-78% decrease in PLTP activity 1032:1077 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 6 61 theme removal 804:810 arg1 effect 764:769 The effect 760:769 The effect of site-specific N-glycosylation removal on PLTP secretion 760:828 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 6 62 theme N-glycosylation 788:802 arg1 removal 804:810 site-specific N-glycosylation removal 774:810 site-specific N-glycosylation removal 774:810 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 7 63 theme PLTP 1117:1120 arg1 activity 1131:1138 PLTP specific activity 1117:1138 PLTP specific activity 1117:1138 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 3 64 theme ovary 486:490 arg1 cells 499:503 Chinese hamster ovary Flp-In cells 470:503 Chinese hamster ovary Flp-In cells 470:503 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 8 65 contain had 1318:1320 arg1 removal 1283:1289 the removal 1279:1289 the removal of the carbohydrate chains 1279:1316 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 8 65 contain had 1318:1320 arg2 impact 1337:1342 a quantitative impact 1322:1342 a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity 1322:1411 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 7 66 theme % 1103:1103 arg1 decrease 1105:1112 a significant 29-80% decrease 1084:1112 a significant 29-80% decrease in PLTP specific activity 1084:1138 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 5 67 from site 727:730 arg1 size 678:681 The size 674:681 The size of the carbohydrate at each N-glycosylation site 674:730 The size of the carbohydrate at each N-glycosylation site ranged from 3.14 to 4.2kDa. 21515415 7 68 gly glycosylation 1000:1012 arg2 sites 1014:1018 the six glycosylation sites 992:1018 the six glycosylation sites 992:1018 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 7 68 gly glycosylation 1000:1012 arg2 six 996:998 six 996:998 six 996:998 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 4 69 gly glycosylated 648:659 arg1 sites 638:642 all six sites 630:642 all six sites 630:642 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 6 70 theme site-specific 774:786 arg1 removal 804:810 site-specific N-glycosylation removal 774:810 site-specific N-glycosylation removal 774:810 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 8 71 theme quantitative 1324:1335 arg1 impact 1337:1342 a quantitative impact 1322:1342 a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity 1322:1411 These data indicate that although no single N-linked carbohydrate chain is a requirement for secretion or activity, the removal of the carbohydrate chains had a quantitative impact on cellular secretion of PLTP and its phospholipid transfer activity. 21515415 3 72 theme Chinese 470:476 arg1 cells 499:503 Chinese hamster ovary Flp-In cells 470:503 Chinese hamster ovary Flp-In cells 470:503 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 6 73 dep enhancement 851:861 arg1 % 866:866 15% 864:866 15% 864:866 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 6 73 dep enhancement 851:861 arg1 % 874:874 60% 872:874 60% 872:874 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 6 73 dep enhancement 851:861 arg1 effect 896:901 no effect 893:901 no effect 893:901 The effect of site-specific N-glycosylation removal on PLTP secretion varied from a modest enhancement (15% and 60%), or essentially no effect, to a reduction in secretion (8%, 14% and 32%). 21515415 3 74 theme sites 314:318 arg1 impact 298:303 the impact 294:303 the impact of these sites on PLTP secretion and activity 294:349 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 4 75 theme Western 600:606 arg1 blot 608:611 Western blot 600:611 Western blot 600:611 The apparent size of each of the six PLTP mutants was slightly less than that of wild type by Western blot, indicating that all six sites are glycosylated or utilized. 21515415 1 76 theme plasma 143:148 arg1 PLTP 181:184 PLTP 181:184 PLTP 181:184 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 1 76 theme plasma 143:148 arg1 protein 172:178 The plasma phospholipid transfer protein 139:178 The plasma phospholipid transfer protein (PLTP) 139:185 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 1 77 theme lipoprotein 217:227 arg1 metabolism 229:238 lipoprotein metabolism 217:238 lipoprotein metabolism 217:238 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 7 78 theme 35-78 1046:1050 arg1 % 1051:1051 % 1051:1051 % 1051:1051 Removal of N-glycosylation at any one of the six glycosylation sites resulted in a significant 35-78% decrease in PLTP activity, and a significant 29-80% decrease in PLTP specific activity compared to wild type. 21515415 1 79 theme phospholipid 150:161 arg1 PLTP 181:184 PLTP 181:184 PLTP 181:184 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 1 79 theme phospholipid 150:161 arg1 protein 172:178 The plasma phospholipid transfer protein 139:178 The plasma phospholipid transfer protein (PLTP) 139:185 The plasma phospholipid transfer protein (PLTP) plays a key role in lipid and lipoprotein metabolism. 21515415 3 80 from impact 298:303 arg1 secretion 328:336 PLTP secretion 323:336 PLTP secretion 323:336 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 21515415 3 80 from impact 298:303 arg1 activity 342:349 activity 342:349 activity 342:349 To study the impact of these sites on PLTP secretion and activity, six variants containing serine to alanine point mutations were prepared by site-directed mutagenesis and expressed in Chinese hamster ovary Flp-In cells. 22750213 3 0 theme PRiMA 603:607 arg1 glycosylation 609:621 PRiMA glycosylation 603:621 PRiMA glycosylation 603:621 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 2 1 from presence 410:417 arg1 AChE 478:481 AChE 478:481 AChE(T) 478:484 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 2 1 from presence 410:417 arg1 subunit 469:475 the AChE catalytic subunit 450:475 the AChE catalytic subunit (AChE(T)) 450:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 6 2 theme mouse 894:898 arg1 PRiMA 900:904 mouse PRiMA 894:904 mouse PRiMA 894:904 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 2 3 theme C-terminal 424:433 arg1 " 445:445 a C-terminal "t-peptide" 422:445 a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)) 422:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 4 4 contain containing 695:704 arg1 PRiMA 671:675 PRiMA 671:675 PRiMA 671:675 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 4 4 contain containing 695:704 arg1 glycoprotein 682:693 a glycoprotein 680:693 a glycoprotein containing two putative N-linked glycosylation sites 680:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 4 4 contain containing 695:704 arg2 sites 742:746 two putative N-linked glycosylation sites 706:746 two putative N-linked glycosylation sites 706:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 2 5 theme catalytic 459:467 arg1 AChE 478:481 AChE 478:481 AChE(T) 478:484 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 2 5 theme catalytic 459:467 arg1 subunit 469:475 the AChE catalytic subunit 450:475 the AChE catalytic subunit (AChE(T)) 450:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 7 6 gly glycoproteins 1167:1179 arg1 glycoproteins 1167:1179 membrane glycoproteins 1158:1179 membrane glycoproteins 1158:1179 This result is contrary to the reports that glycosylation is essential for conformation and trafficking of membrane glycoproteins. 22750213 6 7 theme PRiMA-linked 1022:1033 arg1 tetramers 1040:1048 PRiMA-linked AChE tetramers 1022:1048 PRiMA-linked AChE tetramers 1022:1048 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 5 8 theme glycosylation 837:849 arg1 PRiMA 859:863 PRiMA 859:863 PRiMA 859:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 8 theme glycosylation 837:849 arg1 asparagine-43 789:801 the asparagine-43 785:801 the asparagine-43 785:801 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 8 theme glycosylation 837:849 arg1 site 851:854 the N-linked glycosylation site 824:854 the N-linked glycosylation site of PRiMA 824:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 1 9 theme structural 230:239 arg1 protein 241:247 a structural protein 228:247 a structural protein called proline-rich membrane anchor (PRiMA) 228:291 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 2 10 theme AChE 454:457 arg1 AChE 478:481 AChE 478:481 AChE(T) 478:484 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 2 10 theme AChE 454:457 arg1 subunit 469:475 the AChE catalytic subunit 450:475 the AChE catalytic subunit (AChE(T)) 450:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 1 11 theme cell 316:319 arg1 membrane 321:328 the cell membrane 312:328 the cell membrane of neurons in the brain 312:352 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 6 12 theme tetramers 1040:1048 arg1 assembly 933:940 its assembly 929:940 its assembly with AChE(T) 929:953 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 12 theme tetramers 1040:1048 arg1 trafficking 1007:1017 the membrane trafficking 994:1017 the membrane trafficking of PRiMA-linked AChE tetramers 994:1048 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 12 theme tetramers 1040:1048 arg1 properties 970:979 the enzymatic properties 956:979 the enzymatic properties of AChE 956:987 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 13 theme Abolishing 866:875 arg1 glycosylation 877:889 Abolishing glycosylation 866:889 Abolishing glycosylation on mouse PRiMA 866:904 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 5 14 gly glycosylation 837:849 arg2 PRiMA 859:863 PRiMA 859:863 PRiMA 859:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 14 gly glycosylation 837:849 arg1 PRiMA 859:863 PRiMA 859:863 PRiMA 859:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 14 gly glycosylation 837:849 arg2 site 851:854 the N-linked glycosylation site 824:854 the N-linked glycosylation site of PRiMA 824:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 14 gly glycosylation 837:849 arg2 asparagine-43 789:801 the asparagine-43 785:801 the asparagine-43 785:801 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 4 15 theme putative 710:717 arg1 sites 742:746 two putative N-linked glycosylation sites 706:746 two putative N-linked glycosylation sites 706:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 0 16 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of proline-rich membrane anchor (PRiMA) 0:61 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 5 17 theme PRiMA 859:863 arg1 PRiMA 859:863 PRiMA 859:863 PRiMA 859:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 17 theme PRiMA 859:863 arg1 asparagine-43 789:801 the asparagine-43 785:801 the asparagine-43 785:801 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 17 theme PRiMA 859:863 arg1 site 851:854 the N-linked glycosylation site 824:854 the N-linked glycosylation site of PRiMA 824:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 3 18 gly glycosylation 609:621 arg1 oligomer 630:637 the oligomer assembly 626:646 the oligomer assembly 626:646 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 6 19 link PRiMA-linked 1022:1033 arg1 tetramers 1040:1048 PRiMA-linked AChE tetramers 1022:1048 PRiMA-linked AChE tetramers 1022:1048 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 1 20 theme neurons 333:339 arg1 membrane 321:328 the cell membrane 312:328 the cell membrane of neurons in the brain 312:352 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 4 21 theme glycosylation 728:740 arg1 sites 742:746 two putative N-linked glycosylation sites 706:746 two putative N-linked glycosylation sites 706:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 5 22 link N-linked 828:835 arg1 PRiMA 859:863 PRiMA 859:863 PRiMA 859:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 22 link N-linked 828:835 arg1 asparagine-43 789:801 the asparagine-43 785:801 the asparagine-43 785:801 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 22 link N-linked 828:835 arg1 site 851:854 the N-linked glycosylation site 824:854 the N-linked glycosylation site of PRiMA 824:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 0 23 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of proline-rich membrane anchor (PRiMA) 0:61 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 4 24 theme N-linked 719:726 arg1 sites 742:746 two putative N-linked glycosylation sites 706:746 two putative N-linked glycosylation sites 706:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 6 25 from glycosylation 877:889 arg1 PRiMA 900:904 mouse PRiMA 894:904 mouse PRiMA 894:904 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 0 26 theme proline-rich 26:37 arg1 anchor 48:53 proline-rich membrane anchor 26:53 proline-rich membrane anchor (PRiMA) 26:61 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 0 26 theme proline-rich 26:37 arg1 PRiMA 56:60 PRiMA 56:60 PRiMA 56:60 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 1 27 theme proline-rich 256:267 arg1 PRiMA 286:290 PRiMA 286:290 PRiMA 286:290 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 1 27 theme proline-rich 256:267 arg1 anchor 278:283 proline-rich membrane anchor 256:283 proline-rich membrane anchor (PRiMA) 256:291 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 0 28 theme globular 111:118 arg1 acetylcholinesterase 131:150 globular tetrameric acetylcholinesterase 111:150 globular tetrameric acetylcholinesterase 111:150 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 5 29 theme N-linked 828:835 arg1 PRiMA 859:863 PRiMA 859:863 PRiMA 859:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 29 theme N-linked 828:835 arg1 asparagine-43 789:801 the asparagine-43 785:801 the asparagine-43 785:801 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 5 29 theme N-linked 828:835 arg1 site 851:854 the N-linked glycosylation site 824:854 the N-linked glycosylation site of PRiMA 824:863 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 2 30 theme AChE 371:374 arg1 tetramers 376:384 AChE tetramers 371:384 AChE tetramers 371:384 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 2 31 theme tetramers 376:384 arg1 assembly 359:366 The assembly 355:366 The assembly of AChE tetramers with PRiMA 355:395 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 1 32 theme membrane 269:276 arg1 PRiMA 286:290 PRiMA 286:290 PRiMA 286:290 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 1 32 theme membrane 269:276 arg1 anchor 278:283 proline-rich membrane anchor 256:283 proline-rich membrane anchor (PRiMA) 256:291 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 6 33 theme AChE 1035:1038 arg1 tetramers 1040:1048 PRiMA-linked AChE tetramers 1022:1048 PRiMA-linked AChE tetramers 1022:1048 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 4 34 gly glycoprotein 682:693 arg1 PRiMA 671:675 PRiMA 671:675 PRiMA 671:675 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 4 34 gly glycoprotein 682:693 arg1 glycoprotein 682:693 a glycoprotein 680:693 a glycoprotein containing two putative N-linked glycosylation sites 680:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 1 35 from membrane 321:328 arg1 brain 348:352 the brain 344:352 the brain 344:352 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 0 36 theme anchor 48:53 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of proline-rich membrane anchor (PRiMA) 0:61 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 3 37 theme glycosylation 609:621 arg1 role 595:598 the role 591:598 the role of PRiMA glycosylation in the oligomer assembly 591:646 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 0 38 theme acetylcholinesterase 131:150 arg1 assembly 83:90 assembly 83:90 assembly 83:90 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 0 38 theme acetylcholinesterase 131:150 arg1 trafficking 96:106 trafficking 96:106 trafficking 96:106 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 7 39 theme membrane 1158:1165 arg1 glycoproteins 1167:1179 membrane glycoproteins 1158:1179 membrane glycoproteins 1158:1179 This result is contrary to the reports that glycosylation is essential for conformation and trafficking of membrane glycoproteins. 22750213 7 40 theme glycoproteins 1167:1179 arg1 trafficking 1143:1153 trafficking 1143:1153 trafficking 1143:1153 This result is contrary to the reports that glycosylation is essential for conformation and trafficking of membrane glycoproteins. 22750213 7 40 theme glycoproteins 1167:1179 arg1 conformation 1126:1137 conformation 1126:1137 conformation 1126:1137 This result is contrary to the reports that glycosylation is essential for conformation and trafficking of membrane glycoproteins. 22750213 3 41 gly glycosylation 492:504 arg1 T 514:514 T 514:514 T 514:514 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 3 41 gly glycosylation 492:504 arg1 AChE 509:512 AChE 509:512 AChE(T) 509:515 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 0 42 theme membrane 39:46 arg1 anchor 48:53 proline-rich membrane anchor 26:53 proline-rich membrane anchor (PRiMA) 26:61 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 0 42 theme membrane 39:46 arg1 PRiMA 56:60 PRiMA 56:60 PRiMA 56:60 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 0 43 theme tetrameric 120:129 arg1 acetylcholinesterase 131:150 globular tetrameric acetylcholinesterase 111:150 globular tetrameric acetylcholinesterase 111:150 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 2 44 from " 445:445 arg1 AChE 478:481 AChE 478:481 AChE(T) 478:484 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 2 44 from " 445:445 arg1 subunit 469:475 the AChE catalytic subunit 450:475 the AChE catalytic subunit (AChE(T)) 450:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 0 45 gly glycosylation 9:21 arg1 anchor 48:53 proline-rich membrane anchor 26:53 proline-rich membrane anchor (PRiMA) 26:61 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 0 45 gly glycosylation 9:21 arg1 acetylcholinesterase 131:150 globular tetrameric acetylcholinesterase 111:150 globular tetrameric acetylcholinesterase 111:150 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 0 45 gly glycosylation 9:21 arg1 PRiMA 56:60 PRiMA 56:60 PRiMA 56:60 N-linked glycosylation of proline-rich membrane anchor (PRiMA) is not required for assembly and trafficking of globular tetrameric acetylcholinesterase. 22750213 4 46 gly glycosylation 728:740 arg2 two 706:708 two 706:708 two 706:708 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 4 46 gly glycosylation 728:740 arg2 sites 742:746 two putative N-linked glycosylation sites 706:746 two putative N-linked glycosylation sites 706:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 6 47 with properties 970:979 arg1 T 952:952 T 952:952 T 952:952 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 47 with properties 970:979 arg1 AChE 947:950 AChE 947:950 AChE(T) 947:953 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 7 48 gly glycosylation 1095:1107 arg1 glycoproteins 1167:1179 membrane glycoproteins 1158:1179 membrane glycoproteins 1158:1179 This result is contrary to the reports that glycosylation is essential for conformation and trafficking of membrane glycoproteins. 22750213 3 49 theme oligomer 630:637 arg1 assembly 639:646 the oligomer assembly 626:646 the oligomer assembly 626:646 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 3 50 theme proper 549:554 arg1 assembly 556:563 its proper assembly 545:563 its proper assembly 545:563 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 2 51 with assembly 359:366 arg1 PRiMA 391:395 PRiMA 391:395 PRiMA 391:395 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 6 52 with trafficking 1007:1017 arg1 T 952:952 T 952:952 T 952:952 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 52 with trafficking 1007:1017 arg1 AChE 947:950 AChE 947:950 AChE(T) 947:953 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 3 53 theme AChE 509:512 arg1 glycosylation 492:504 The glycosylation 488:504 The glycosylation of AChE(T) 488:515 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 22750213 6 54 theme AChE 984:987 arg1 assembly 933:940 its assembly 929:940 its assembly with AChE(T) 929:953 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 54 theme AChE 984:987 arg1 trafficking 1007:1017 the membrane trafficking 994:1017 the membrane trafficking of PRiMA-linked AChE tetramers 994:1048 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 54 theme AChE 984:987 arg1 properties 970:979 the enzymatic properties 956:979 the enzymatic properties of AChE 956:987 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 55 theme membrane 998:1005 arg1 trafficking 1007:1017 the membrane trafficking 994:1017 the membrane trafficking of PRiMA-linked AChE tetramers 994:1048 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 56 gly glycosylation 877:889 arg1 PRiMA 900:904 mouse PRiMA 894:904 mouse PRiMA 894:904 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 2 57 from subunit 469:475 arg1 presence 410:417 the presence 406:417 the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)) 406:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 2 58 theme " 445:445 arg1 presence 410:417 the presence 406:417 the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)) 406:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 6 59 with assembly 933:940 arg1 T 952:952 T 952:952 T 952:952 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 6 59 with assembly 933:940 arg1 AChE 947:950 AChE 947:950 AChE(T) 947:953 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 1 60 theme globular 199:206 arg1 tetramers 208:216 globular tetramers 199:216 globular tetramers (G(4)) 199:223 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 5 61 theme site-directed 758:770 arg1 mutagenesis 772:782 site-directed mutagenesis 758:782 site-directed mutagenesis 758:782 By using site-directed mutagenesis, the asparagine-43 was identified to be the N-linked glycosylation site of PRiMA. 22750213 2 62 theme t-peptide 436:444 arg1 " 445:445 a C-terminal "t-peptide" 422:445 a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)) 422:485 The assembly of AChE tetramers with PRiMA requires the presence of a C-terminal "t-peptide" in the AChE catalytic subunit (AChE(T)). 22750213 1 63 dep tetramers 208:216 arg1 4 221:221 4 221:221 4 221:221 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 1 63 dep tetramers 208:216 arg1 G 219:219 G 219:219 G(4) 219:222 Acetylcholinesterase (AChE) is organized into globular tetramers (G(4)) by a structural protein called proline-rich membrane anchor (PRiMA), anchoring it into the cell membrane of neurons in the brain. 22750213 6 64 theme enzymatic 960:968 arg1 properties 970:979 the enzymatic properties 956:979 the enzymatic properties of AChE 956:987 Abolishing glycosylation on mouse PRiMA appeared not to affect its assembly with AChE(T), the enzymatic properties of AChE, and the membrane trafficking of PRiMA-linked AChE tetramers. 22750213 4 65 link N-linked 719:726 arg1 sites 742:746 two putative N-linked glycosylation sites 706:746 two putative N-linked glycosylation sites 706:746 PRiMA is a glycoprotein containing two putative N-linked glycosylation sites. 22750213 3 66 from role 595:598 arg1 assembly 639:646 the oligomer assembly 626:646 the oligomer assembly 626:646 The glycosylation of AChE(T) is known to be required for its proper assembly and trafficking; however, the role of PRiMA glycosylation in the oligomer assembly has not been revealed. 26274980 3 0 theme intact 1030:1035 arg1 nCG 1037:1039 intact nCG 1030:1039 intact nCG 1030:1039 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 2 1 theme Manα1,6Manβ1,4GlcNAcβ1,4 859:882 arg1 GlcNAcβ 892:898 Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 859:898 Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 859:898 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 5 2 theme N-glycosylation 1652:1666 arg1 analysis 1617:1624 the analysis 1613:1624 the analysis of unconventional protein N-glycosylation 1613:1666 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 4 3 theme Asn71-glycans 1312:1324 arg1 nature 1302:1307 the truncated nature 1288:1307 the truncated nature of Asn71-glycans (~5-17 Å) 1288:1334 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 0 4 theme central 201:207 arg1 N-Glycan 29:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 4 theme central 201:207 arg1 N-Glycopeptide 39:52 N-Glycopeptide 39:52 N-Glycopeptide 39:52 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 4 theme central 201:207 arg1 protease 216:223 a central serine protease 199:223 a central serine protease in the human innate immune system 199:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 4 theme central 201:207 arg1 Profiling 81:89 Intact N-Glycoprotein Profiling 59:89 Intact N-Glycoprotein Profiling 59:89 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 4 5 theme proteolytic 1381:1391 arg1 activity 1393:1400 the proteolytic activity 1377:1400 the proteolytic activity of the protein 1377:1415 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 3 6 theme C-terminal 1093:1102 arg1 both 1162:1165 both 1162:1165 both 1162:1165 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 3 6 theme C-terminal 1093:1102 arg1 variants 1115:1122 two C-terminal truncation variants 1089:1122 two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%) 1089:1159 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 0 7 theme Cathepsin 154:162 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 7 theme Cathepsin 154:162 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 1 8 theme N-glycan 407:414 arg1 N-glycopeptide 417:430 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 N-glycopeptide 417:430 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 3 9 from Ser244 1146:1151 arg1 both 1162:1165 both 1162:1165 both 1162:1165 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 3 9 from Ser244 1146:1151 arg1 variants 1115:1122 two C-terminal truncation variants 1089:1122 two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%) 1089:1159 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 6 10 theme Asn71-glycosylation 1753:1771 arg1 role 1741:1744 the functional role 1726:1744 the functional role of nCG Asn71-glycosylation 1726:1771 Importantly, this study now facilitates investigation of the functional role of nCG Asn71-glycosylation. 26274980 4 11 theme direct 1356:1361 arg1 modulation 1363:1372 their direct modulation 1350:1372 their direct modulation of the proteolytic activity of the protein 1350:1415 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 0 12 theme Neutrophil 167:176 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 12 theme Neutrophil 167:176 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 13 theme N-glycosylation 286:300 arg1 importance 268:277 the importance 264:277 the importance of its N-glycosylation 264:300 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 3 14 from Arg243 1127:1132 arg1 both 1162:1165 both 1162:1165 both 1162:1165 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 3 14 from Arg243 1127:1132 arg1 variants 1115:1122 two C-terminal truncation variants 1089:1122 two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%) 1089:1159 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 2 15 theme chitobiose 647:656 arg1 core 658:661 truncated chitobiose core 637:661 truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%) 637:701 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 0 16 theme G 188:188 arg1 Asn71-Glycosylation 114:132 Unconventional Asn71-Glycosylation 99:132 Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 99:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 2 17 gly α2,3-sialylated 946:960 arg1 N-glycans 970:978 α2,3-sialylated complex N-glycans 946:978 α2,3-sialylated complex N-glycans 946:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 6 18 theme functional 1730:1739 arg1 role 1741:1744 the functional role 1726:1744 the functional role of nCG Asn71-glycosylation 1726:1771 Importantly, this study now facilitates investigation of the functional role of nCG Asn71-glycosylation. 26274980 1 19 theme glycoprotein 444:455 arg1 profiling 457:465 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 profiling 457:465 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 0 20 theme N-Glycoprotein 66:79 arg1 N-Glycan 29:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 20 theme N-Glycoprotein 66:79 arg1 N-Glycopeptide 39:52 N-Glycopeptide 39:52 N-Glycopeptide 39:52 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 20 theme N-Glycoprotein 66:79 arg1 Profiling 81:89 Intact N-Glycoprotein Profiling 59:89 Intact N-Glycoprotein Profiling 59:89 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 20 theme N-Glycoprotein 66:79 arg1 protease 216:223 a central serine protease 199:223 a central serine protease in the human innate immune system 199:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 1 21 theme such 345:348 arg1 investigations 350:363 such investigations 345:363 such investigations 345:363 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 4 22 theme protein 1409:1415 arg1 activity 1393:1400 the proteolytic activity 1377:1400 the proteolytic activity of the protein 1377:1415 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 0 23 theme immune 245:250 arg1 system 252:257 the human innate immune system 228:257 the human innate immune system 228:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 1 24 dep micro- 495:500 arg1 the 491:493 the 491:493 the 491:493 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 2 25 theme occupied 569:576 arg1 Asn71 578:582 The fully occupied Asn71 559:582 The fully occupied Asn71 559:582 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 1 26 gly macro-heterogeneity 506:524 arg1 nCG 529:531 nCG 529:531 nCG 529:531 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 2 27 contain carried 584:590 arg1 Asn71 578:582 The fully occupied Asn71 559:582 The fully occupied Asn71 559:582 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 27 contain carried 584:590 arg2 N-glycosylation 607:621 unconventional N-glycosylation 592:621 unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans 592:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 0 28 theme Human 137:141 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 28 theme Human 137:141 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 1 29 theme complementary 378:390 arg1 N-glycopeptide 417:430 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 N-glycopeptide 417:430 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 0 30 theme Complementary 0:12 arg1 N-Glycan 29:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 30 theme Complementary 0:12 arg1 N-Glycopeptide 39:52 N-Glycopeptide 39:52 N-Glycopeptide 39:52 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 30 theme Complementary 0:12 arg1 protease 216:223 a central serine protease 199:223 a central serine protease in the human innate immune system 199:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 30 theme Complementary 0:12 arg1 Profiling 81:89 Intact N-Glycoprotein Profiling 59:89 Intact N-Glycoprotein Profiling 59:89 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 2 31 theme α2,6- 936:940 arg1 core 658:661 truncated chitobiose core 637:661 truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%) 637:701 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 31 theme α2,6- 936:940 arg1 N-glycans 720:728 paucimannosidic N-glycans 704:728 paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ) 704:899 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 31 theme α2,6- 936:940 arg1 levels 912:917 trace levels 906:917 trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans 906:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 32 dep % 761:761 arg1 Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ 805:835 Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ 805:835 Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ 805:835 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 32 dep % 761:761 arg1 GlcNAcβ 790:796 Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 764:796 Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 764:796 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 32 dep % 761:761 arg1 % 802:802 7.9% 799:802 7.9% 799:802 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 32 dep % 761:761 arg1 % 841:841 3.7% 838:841 3.7% 838:841 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 1 33 from individuals 546:556 arg1 macro-heterogeneity 506:524 macro-heterogeneity 506:524 macro-heterogeneity 506:524 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 1 33 from individuals 546:556 arg1 micro- 495:500 micro- 495:500 micro- 495:500 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 5 34 theme important 1576:1584 arg1 challenges 1586:1595 important challenges 1576:1595 important challenges associated with the analysis of unconventional protein N-glycosylation 1576:1666 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 3 35 theme LC-MS 1011:1015 arg1 profiling 1017:1025 High-resolution/mass accuracy LC-MS profiling 981:1025 High-resolution/mass accuracy LC-MS profiling of intact nCG 981:1039 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 2 36 theme N-glycans 970:978 arg1 core 658:661 truncated chitobiose core 637:661 truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%) 637:701 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 36 theme N-glycans 970:978 arg1 N-glycans 720:728 paucimannosidic N-glycans 704:728 paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ) 704:899 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 36 theme N-glycans 970:978 arg1 levels 912:917 trace levels 906:917 trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans 906:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 4 37 theme truncated 1292:1300 arg1 nature 1302:1307 the truncated nature 1288:1307 the truncated nature of Asn71-glycans (~5-17 Å) 1288:1334 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 3 38 theme High-resolution/mass 981:1000 arg1 profiling 1017:1025 High-resolution/mass accuracy LC-MS profiling 981:1025 High-resolution/mass accuracy LC-MS profiling of intact nCG 981:1039 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 2 39 theme trace 844:848 arg1 % 761:761 10.6% 757:761 10.6% 757:761 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 39 theme trace 844:848 arg1 level 850:854 trace level 844:854 trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 844:898 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 40 theme α2,3-sialylated 946:960 arg1 N-glycans 970:978 α2,3-sialylated complex N-glycans 946:978 α2,3-sialylated complex N-glycans 946:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 5 41 theme simple 1539:1544 arg1 glycoproteins 1546:1558 even relatively simple glycoproteins 1523:1558 even relatively simple glycoproteins 1523:1558 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 4 42 theme nCG 1272:1274 arg1 nCG 1272:1274 nCG 1272:1274 nCG 1272:1274 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 4 42 theme nCG 1272:1274 arg1 site 1264:1267 the active site 1253:1267 the active site of nCG 1253:1274 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 0 43 dep N-Glycan 29:36 arg1 Reveals 91:97 Reveals 91:97 Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 91:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 3 44 theme nCG 1037:1039 arg1 profiling 1017:1025 High-resolution/mass accuracy LC-MS profiling 981:1025 High-resolution/mass accuracy LC-MS profiling of intact nCG 981:1039 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 2 45 theme GlcNAcβ 892:898 arg1 % 761:761 10.6% 757:761 10.6% 757:761 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 45 theme GlcNAcβ 892:898 arg1 level 850:854 trace level 844:854 trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 844:898 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 1 46 gly N-glycopeptide 417:430 arg2 N-glycopeptide 417:430 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 N-glycopeptide 417:430 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 2 47 theme Fucα1,6 884:890 arg1 GlcNAcβ 892:898 Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 859:898 Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 859:898 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 5 48 theme protein 1644:1650 arg1 N-glycosylation 1652:1666 unconventional protein N-glycosylation 1629:1666 unconventional protein N-glycosylation 1629:1666 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 5 49 gly glycoproteins 1546:1558 arg1 glycoproteins 1546:1558 even relatively simple glycoproteins 1523:1558 even relatively simple glycoproteins 1523:1558 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 3 50 theme solvent-accessible 1191:1208 arg1 Met152 1210:1215 solvent-accessible Met152 1191:1215 solvent-accessible Met152 1191:1215 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 2 51 theme trace 906:910 arg1 levels 912:917 trace levels 906:917 trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans 906:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 0 52 theme serine 209:214 arg1 N-Glycan 29:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 52 theme serine 209:214 arg1 N-Glycopeptide 39:52 N-Glycopeptide 39:52 N-Glycopeptide 39:52 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 52 theme serine 209:214 arg1 protease 216:223 a central serine protease 199:223 a central serine protease in the human innate immune system 199:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 52 theme serine 209:214 arg1 Profiling 81:89 Intact N-Glycoprotein Profiling 59:89 Intact N-Glycoprotein Profiling 59:89 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 1 53 gly glycoprotein 444:455 arg1 glycoprotein 444:455 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 glycoprotein 444:455 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 5 54 theme continued 1443:1451 arg1 requirement 1453:1463 the continued requirement 1439:1463 the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins 1439:1558 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 3 55 theme truncation 1104:1113 arg1 both 1162:1165 both 1162:1165 both 1162:1165 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 3 55 theme truncation 1104:1113 arg1 variants 1115:1122 two C-terminal truncation variants 1089:1122 two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%) 1089:1159 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 0 56 theme human 232:236 arg1 system 252:257 the human innate immune system 228:257 the human innate immune system 228:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 57 theme G. 164:165 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 57 theme G. 164:165 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 4 58 dep proximal 1233:1240 arg1 Å 1247:1247 ~19 Å 1243:1247 ~19 Å 1243:1247 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 2 59 theme unconventional 592:605 arg1 N-glycosylation 607:621 unconventional N-glycosylation 592:621 unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans 592:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 4 60 theme activity 1393:1400 arg1 modulation 1363:1372 their direct modulation 1350:1372 their direct modulation of the proteolytic activity of the protein 1350:1415 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 0 61 theme cathepsin 178:186 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 61 theme cathepsin 178:186 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 2 62 dep core 658:661 arg1 GlcNAcβ 664:670 GlcNAcβ 664:670 GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7% 664:700 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 6 63 theme nCG 1749:1751 arg1 Asn71-glycosylation 1753:1771 nCG Asn71-glycosylation 1749:1771 nCG Asn71-glycosylation 1749:1771 Importantly, this study now facilitates investigation of the functional role of nCG Asn71-glycosylation. 26274980 0 64 from protease 216:223 arg1 system 252:257 the human innate immune system 228:257 the human innate immune system 228:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 2 65 dep GlcNAcβ 664:670 arg1 % 700:700 22.7% 696:700 GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7% 664:700 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 65 dep GlcNAcβ 664:670 arg1 % 677:677 55.2% 673:677 GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7% 664:700 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 65 dep GlcNAcβ 664:670 arg1 Fucα1,6GlcNAcβ 680:693 Fucα1,6GlcNAcβ 680:693 GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7% 664:700 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 66 theme truncated 637:645 arg1 core 658:661 truncated chitobiose core 637:661 truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%) 637:701 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 6 67 theme role 1741:1744 arg1 investigation 1709:1721 investigation 1709:1721 investigation of the functional role of nCG Asn71-glycosylation 1709:1771 Importantly, this study now facilitates investigation of the functional role of nCG Asn71-glycosylation. 26274980 1 68 theme intact 437:442 arg1 profiling 457:465 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 profiling 457:465 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 2 69 theme Fucα1,6 782:788 arg1 GlcNAcβ 790:796 Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 764:796 Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 764:796 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 70 dep N-glycans 720:728 arg1 Manβ1,4GlcNAcβ1,4GlcNAcβ 731:754 Manβ1,4GlcNAcβ1,4GlcNAcβ 731:754 Manβ1,4GlcNAcβ1,4GlcNAcβ 731:754 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 71 theme Manβ1,4GlcNAcβ1,4 764:780 arg1 GlcNAcβ 790:796 Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 764:796 Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 764:796 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 0 72 theme Unconventional 99:112 arg1 Asn71-Glycosylation 114:132 Unconventional Asn71-Glycosylation 99:132 Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 99:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 4 73 theme active 1257:1262 arg1 nCG 1272:1274 nCG 1272:1274 nCG 1272:1274 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 4 73 theme active 1257:1262 arg1 site 1264:1267 the active site 1253:1267 the active site of nCG 1253:1274 Asn71 appeared proximal (~19 Å) to the active site of nCG, but due to the truncated nature of Asn71-glycans (~5-17 Å) we questioned their direct modulation of the proteolytic activity of the protein. 26274980 0 74 theme innate 238:243 arg1 system 252:257 the human innate immune system 228:257 the human innate immune system 228:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 75 theme Neutrophil 143:152 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 75 theme Neutrophil 143:152 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 76 theme LC-MS/MS-Based 14:27 arg1 N-Glycan 29:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 76 theme LC-MS/MS-Based 14:27 arg1 N-Glycopeptide 39:52 N-Glycopeptide 39:52 N-Glycopeptide 39:52 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 76 theme LC-MS/MS-Based 14:27 arg1 protease 216:223 a central serine protease 199:223 a central serine protease in the human innate immune system 199:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 76 theme LC-MS/MS-Based 14:27 arg1 Profiling 81:89 Intact N-Glycoprotein Profiling 59:89 Intact N-Glycoprotein Profiling 59:89 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 1 77 theme LC-MS/MS-based 392:405 arg1 N-glycopeptide 417:430 complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling 378:465 N-glycopeptide 417:430 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 0 78 gly Asn71-Glycosylation 114:132 arg1 nCG 191:193 nCG 191:193 nCG 191:193 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 78 gly Asn71-Glycosylation 114:132 arg1 G 188:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G 137:188 Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) 137:194 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 5 79 theme complementary 1474:1486 arg1 technologies 1488:1499 complementary technologies 1474:1499 complementary technologies 1474:1499 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 26274980 2 80 theme paucimannosidic 704:718 arg1 N-glycans 720:728 paucimannosidic N-glycans 704:728 paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ) 704:899 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 81 dep Manβ1,4GlcNAcβ1,4GlcNAcβ 731:754 arg1 % 761:761 10.6% 757:761 10.6% 757:761 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 81 dep Manβ1,4GlcNAcβ1,4GlcNAcβ 731:754 arg1 level 850:854 trace level 844:854 trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ 844:898 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 2 82 theme monoantennary 922:934 arg1 α2,6- 936:940 monoantennary α2,6- 922:940 monoantennary α2,6- 922:940 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 1 83 theme nCG 529:531 arg1 macro-heterogeneity 506:524 macro-heterogeneity 506:524 macro-heterogeneity 506:524 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 1 83 theme nCG 529:531 arg1 micro- 495:500 micro- 495:500 micro- 495:500 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 0 84 theme Intact 59:64 arg1 N-Glycan 29:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan 0:36 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 84 theme Intact 59:64 arg1 N-Glycopeptide 39:52 N-Glycopeptide 39:52 N-Glycopeptide 39:52 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 84 theme Intact 59:64 arg1 Profiling 81:89 Intact N-Glycoprotein Profiling 59:89 Intact N-Glycoprotein Profiling 59:89 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 0 84 theme Intact 59:64 arg1 protease 216:223 a central serine protease 199:223 a central serine protease in the human innate immune system 199:257 Complementary LC-MS/MS-Based N-Glycan, N-Glycopeptide, and Intact N-Glycoprotein Profiling Reveals Unconventional Asn71-Glycosylation of Human Neutrophil Cathepsin G. Neutrophil cathepsin G (nCG) is a central serine protease in the human innate immune system, but the importance of its N-glycosylation remains largely undescribed. 26274980 2 85 theme complex 962:968 arg1 N-glycans 970:978 α2,3-sialylated complex N-glycans 946:978 α2,3-sialylated complex N-glycans 946:978 The fully occupied Asn71 carried unconventional N-glycosylation consisting of truncated chitobiose core (GlcNAcβ: 55.2%; Fucα1,6GlcNAcβ: 22.7%), paucimannosidic N-glycans (Manβ1,4GlcNAcβ1,4GlcNAcβ: 10.6%; Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ: 7.9%; Manα1,6Manβ1,4GlcNAcβ1,4GlcNAcβ: 3.7%, trace level of Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ), and trace levels of monoantennary α2,6- and α2,3-sialylated complex N-glycans. 26274980 1 86 theme healthy 538:544 arg1 individuals 546:556 healthy individuals 538:556 healthy individuals 538:556 To facilitate such investigations, we here use complementary LC-MS/MS-based N-glycan, N-glycopeptide, and intact glycoprotein profiling to accurately establish the micro- and macro-heterogeneity of nCG from healthy individuals. 26274980 3 87 theme accuracy 1002:1009 arg1 profiling 1017:1025 High-resolution/mass accuracy LC-MS profiling 981:1025 High-resolution/mass accuracy LC-MS profiling of intact nCG 981:1039 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 3 88 theme Met152 1210:1215 arg1 oxidation 1178:1186 oxidation 1178:1186 oxidation of solvent-accessible Met152 1178:1215 High-resolution/mass accuracy LC-MS profiling of intact nCG confirmed the Asn71-glycoprofile and identified two C-terminal truncation variants at Arg243 (57.8%) and Ser244 (42.2%), both displaying oxidation of solvent-accessible Met152. 26274980 5 89 theme unconventional 1629:1642 arg1 N-glycosylation 1652:1666 unconventional protein N-glycosylation 1629:1666 unconventional protein N-glycosylation 1629:1666 This work highlights the continued requirement of using complementary technologies to accurately profile even relatively simple glycoproteins and illustrates important challenges associated with the analysis of unconventional protein N-glycosylation. 23050552 5 0 theme detailed 584:591 arg1 analysis 593:600 the detailed analysis 580:600 the detailed analysis of the protein's O-glycosylation 580:633 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 5 1 dep one- 679:682 arg1 -MS 728:730 nanoLC-ESI-ion trap (IT)-MS 704:730 nanoLC-ESI-ion trap (IT)-MS 704:730 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 2 2 theme tissue 298:303 arg1 injury 305:310 tissue injury 298:310 tissue injury 298:310 The protein plays a crucial role in protecting the vascular network against the loss of blood after tissue injury. 23050552 4 3 theme O-linked 440:447 arg1 oligosaccharides 449:464 O-linked oligosaccharides 440:464 O-linked oligosaccharides 440:464 So far O-linked oligosaccharides have rarely been described. 23050552 5 4 theme tryptic- 515:522 arg1 use 508:510 use 508:510 use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation 508:633 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 1 5 theme paired 157:162 arg1 sets 164:167 paired sets 157:167 paired sets of three subunits (α, β, γ) 157:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 5 theme paired 157:162 arg1 subunits 178:185 three subunits 172:185 three subunits (α, β, γ) 172:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 9 6 theme monosialylated 1214:1227 arg1 T-antigen 1229:1237 a monosialylated T-antigen 1212:1237 a monosialylated T-antigen 1212:1237 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 7 7 theme O-glycopeptides 910:924 arg1 spectra 885:891 MS3 spectra 881:891 MS3 spectra of the generated O-glycopeptides 881:924 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 3 8 theme gamma 326:330 arg1 subunits 332:339 The beta and gamma subunits 313:339 The beta and gamma subunits each 313:344 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 7 9 theme backbone 958:965 arg1 cleavages 933:941 cleavages 933:941 cleavages of the peptide backbone 933:965 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 5 10 theme one- 679:682 arg1 approaches 732:741 both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches 674:741 both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches 674:741 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 6 11 theme backbone 861:868 arg1 cleavages 870:878 some peptide backbone cleavages 848:878 some peptide backbone cleavages 848:878 Glycopeptides were analyzed by ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages. 23050552 10 12 theme clotting 1332:1339 arg1 process 1341:1347 the blood clotting process 1322:1347 the blood clotting process 1322:1347 Our findings may help to improve the general understanding of human fibrinogen in the blood clotting process. 23050552 5 13 theme proteinase 528:537 arg1 glycopeptides 562:574 proteinase K-generated fibrinogen glycopeptides 528:574 proteinase K-generated fibrinogen glycopeptides 528:574 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 2 14 theme blood 286:290 arg1 loss 278:281 the loss 274:281 the loss of blood after tissue injury 274:310 The protein plays a crucial role in protecting the vascular network against the loss of blood after tissue injury. 23050552 3 15 dep subunits 332:339 arg1 each 341:344 each 341:344 each 341:344 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 3 16 contain contain 346:352 arg2 site 374:377 one N-glycosylation site 354:377 one N-glycosylation site 354:377 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 3 16 contain contain 346:352 arg1 subunits 332:339 The beta and gamma subunits 313:339 The beta and gamma subunits each 313:344 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 1 17 dep kDa 112:114 arg1 soluble 117:123 soluble 117:123 soluble 117:123 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 10 18 theme general 1277:1283 arg1 understanding 1285:1297 the general understanding 1273:1297 the general understanding of human fibrinogen in the blood clotting process 1273:1347 Our findings may help to improve the general understanding of human fibrinogen in the blood clotting process. 23050552 5 19 theme two-dimensional 688:702 arg1 approaches 732:741 both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches 674:741 both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches 674:741 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 5 20 theme glycopeptides 562:574 arg1 use 508:510 use 508:510 use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation 508:633 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 7 21 theme generated 900:908 arg1 O-glycopeptides 910:924 the generated O-glycopeptides 896:924 the generated O-glycopeptides 896:924 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 1 22 theme Human 86:90 arg1 glycoprotein 132:143 a 340 kDa, soluble plasma glycoprotein 106:143 a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ) 106:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 22 theme Human 86:90 arg1 fibrinogen 92:101 Human fibrinogen 86:101 Human fibrinogen 86:101 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 3 23 theme biantennary 411:421 arg1 N-glycan 423:430 a biantennary N-glycan 409:430 a biantennary N-glycan 409:430 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 9 24 gly monosialylated 1214:1227 arg1 T-antigen 1229:1237 a monosialylated T-antigen 1212:1237 a monosialylated T-antigen 1212:1237 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 8 25 theme attachment 1060:1069 arg1 sites 1071:1075 The previously reported N-glycan attachment sites 1027:1075 The previously reported N-glycan attachment sites of human fibrinogen 1027:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 8 25 theme attachment 1060:1069 arg1 fibrinogen 1086:1095 human fibrinogen 1080:1095 human fibrinogen 1080:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 5 26 theme nanoLC-ESI-ion 704:717 arg1 -MS 728:730 nanoLC-ESI-ion trap (IT)-MS 704:730 nanoLC-ESI-ion trap (IT)-MS 704:730 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 6 27 theme peptide 853:859 arg1 cleavages 870:878 some peptide backbone cleavages 848:878 some peptide backbone cleavages 848:878 Glycopeptides were analyzed by ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages. 23050552 3 28 theme N-glycosylation 358:372 arg1 site 374:377 one N-glycosylation site 354:377 one N-glycosylation site 354:377 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 0 29 theme Glycoproteomic 0:13 arg1 analysis 15:22 Glycoproteomic analysis 0:22 Glycoproteomic analysis of human fibrinogen 0:42 Glycoproteomic analysis of human fibrinogen reveals novel regions of O-glycosylation. 23050552 4 30 link O-linked 440:447 arg1 oligosaccharides 449:464 O-linked oligosaccharides 440:464 O-linked oligosaccharides 440:464 So far O-linked oligosaccharides have rarely been described. 23050552 5 31 dep -MS 728:730 arg1 IT 725:726 IT 725:726 IT 725:726 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 5 32 theme fibrinogen 551:560 arg1 glycopeptides 562:574 proteinase K-generated fibrinogen glycopeptides 528:574 proteinase K-generated fibrinogen glycopeptides 528:574 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 1 33 theme subunits 178:185 arg1 sets 164:167 paired sets 157:167 paired sets of three subunits (α, β, γ) 157:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 33 theme subunits 178:185 arg1 subunits 178:185 three subunits 172:185 three subunits (α, β, γ) 172:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 0 34 theme human 27:31 arg1 fibrinogen 33:42 human fibrinogen 27:42 human fibrinogen 27:42 Glycoproteomic analysis of human fibrinogen reveals novel regions of O-glycosylation. 23050552 5 35 theme K-generated 539:549 arg1 glycopeptides 562:574 proteinase K-generated fibrinogen glycopeptides 528:574 proteinase K-generated fibrinogen glycopeptides 528:574 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 2 36 theme vascular 249:256 arg1 network 258:264 the vascular network 245:264 the vascular network 245:264 The protein plays a crucial role in protecting the vascular network against the loss of blood after tissue injury. 23050552 10 37 theme fibrinogen 1308:1317 arg1 understanding 1285:1297 the general understanding 1273:1297 the general understanding of human fibrinogen in the blood clotting process 1273:1347 Our findings may help to improve the general understanding of human fibrinogen in the blood clotting process. 23050552 5 38 gly glycopeptides 562:574 arg2 glycopeptides 562:574 proteinase K-generated fibrinogen glycopeptides 528:574 proteinase K-generated fibrinogen glycopeptides 528:574 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 3 39 theme beta 317:320 arg1 subunits 332:339 The beta and gamma subunits 313:339 The beta and gamma subunits each 313:344 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 10 40 theme blood 1326:1330 arg1 process 1341:1347 the blood clotting process 1322:1347 the blood clotting process 1322:1347 Our findings may help to improve the general understanding of human fibrinogen in the blood clotting process. 23050552 6 41 theme linkages 835:842 arg1 cleavages 870:878 some peptide backbone cleavages 848:878 some peptide backbone cleavages 848:878 Glycopeptides were analyzed by ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages. 23050552 6 41 theme linkages 835:842 arg1 fragmentations 806:819 fragmentations 806:819 fragmentations of glycosidic linkages 806:842 Glycopeptides were analyzed by ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages. 23050552 1 42 theme 340 108:110 arg1 kDa 112:114 kDa 112:114 kDa 112:114 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 10 43 theme human 1302:1306 arg1 fibrinogen 1308:1317 human fibrinogen 1302:1317 human fibrinogen 1302:1317 Our findings may help to improve the general understanding of human fibrinogen in the blood clotting process. 23050552 7 44 from information 990:1000 arg1 sequence 1017:1024 the peptide sequence 1005:1024 the peptide sequence 1005:1024 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 8 45 theme reported 1042:1049 arg1 sites 1071:1075 The previously reported N-glycan attachment sites 1027:1075 The previously reported N-glycan attachment sites of human fibrinogen 1027:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 8 45 theme reported 1042:1049 arg1 fibrinogen 1086:1095 human fibrinogen 1080:1095 human fibrinogen 1080:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 8 46 theme fibrinogen 1086:1095 arg1 sites 1071:1075 The previously reported N-glycan attachment sites 1027:1075 The previously reported N-glycan attachment sites of human fibrinogen 1027:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 8 46 theme fibrinogen 1086:1095 arg1 fibrinogen 1086:1095 human fibrinogen 1080:1095 human fibrinogen 1080:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 6 47 theme glycosidic 824:833 arg1 linkages 835:842 glycosidic linkages 824:842 glycosidic linkages 824:842 Glycopeptides were analyzed by ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages. 23050552 0 48 theme fibrinogen 33:42 arg1 analysis 15:22 Glycoproteomic analysis 0:22 Glycoproteomic analysis of human fibrinogen 0:42 Glycoproteomic analysis of human fibrinogen reveals novel regions of O-glycosylation. 23050552 8 49 theme human 1080:1084 arg1 fibrinogen 1086:1095 human fibrinogen 1080:1095 human fibrinogen 1080:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 1 50 dep subunits 178:185 arg1 α 188:188 α 188:188 α 188:188 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 50 dep subunits 178:185 arg1 γ 194:194 γ 194:194 γ 194:194 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 50 dep subunits 178:185 arg1 β 191:191 β 191:191 β 191:191 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 10 51 from understanding 1285:1297 arg1 process 1341:1347 the blood clotting process 1322:1347 the blood clotting process 1322:1347 Our findings may help to improve the general understanding of human fibrinogen in the blood clotting process. 23050552 7 52 theme peptide 1009:1015 arg1 sequence 1017:1024 the peptide sequence 1005:1024 the peptide sequence 1005:1024 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 1 53 theme kDa 112:114 arg1 fibrinogen 92:101 Human fibrinogen 86:101 Human fibrinogen 86:101 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 53 theme kDa 112:114 arg1 glycoprotein 132:143 a 340 kDa, soluble plasma glycoprotein 106:143 a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ) 106:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 0 54 theme novel 52:56 arg1 regions 58:64 novel regions 52:64 novel regions of O-glycosylation 52:83 Glycoproteomic analysis of human fibrinogen reveals novel regions of O-glycosylation. 23050552 1 55 gly glycoprotein 132:143 arg1 fibrinogen 92:101 Human fibrinogen 86:101 Human fibrinogen 86:101 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 55 gly glycoprotein 132:143 arg1 glycoprotein 132:143 a 340 kDa, soluble plasma glycoprotein 106:143 a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ) 106:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 7 56 theme essential 980:988 arg1 information 990:1000 essential information 980:1000 essential information on the peptide sequence 980:1024 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 8 57 theme N-glycan 1051:1058 arg1 sites 1071:1075 The previously reported N-glycan attachment sites 1027:1075 The previously reported N-glycan attachment sites of human fibrinogen 1027:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 8 57 theme N-glycan 1051:1058 arg1 fibrinogen 1086:1095 human fibrinogen 1080:1095 human fibrinogen 1080:1095 The previously reported N-glycan attachment sites of human fibrinogen could be confirmed. 23050552 7 58 theme MS3 881:883 arg1 spectra 885:891 MS3 spectra 881:891 MS3 spectra of the generated O-glycopeptides 881:924 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 5 59 theme O-glycosylation 619:633 arg1 analysis 593:600 the detailed analysis 580:600 the detailed analysis of the protein's O-glycosylation 580:633 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 9 60 theme human 1178:1182 arg1 fibrinogen 1184:1193 human fibrinogen 1178:1193 human fibrinogen 1178:1193 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 3 61 used occupied 397:404 arg2 each 380:383 each 380:383 each 380:383 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 3 61 used occupied 397:404 arg2 site 374:377 one N-glycosylation site 354:377 one N-glycosylation site 354:377 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 7 62 gly O-glycopeptides 910:924 arg2 O-glycopeptides 910:924 the generated O-glycopeptides 896:924 the generated O-glycopeptides 896:924 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 23050552 2 63 theme crucial 218:224 arg1 role 226:229 a crucial role 216:229 a crucial role 216:229 The protein plays a crucial role in protecting the vascular network against the loss of blood after tissue injury. 23050552 9 64 theme O-glycosylation 1151:1165 arg1 all 1196:1198 all 1196:1198 all 1196:1198 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 9 64 theme O-glycosylation 1151:1165 arg1 regions 1167:1173 seven novel O-glycosylation regions 1139:1173 seven novel O-glycosylation regions in human fibrinogen 1139:1193 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 1 65 theme plasma 125:130 arg1 fibrinogen 92:101 Human fibrinogen 86:101 Human fibrinogen 86:101 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 1 65 theme plasma 125:130 arg1 glycoprotein 132:143 a 340 kDa, soluble plasma glycoprotein 106:143 a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ) 106:195 Human fibrinogen is a 340 kDa, soluble plasma glycoprotein composed of paired sets of three subunits (α, β, γ). 23050552 0 66 theme O-glycosylation 69:83 arg1 regions 58:64 novel regions 52:64 novel regions of O-glycosylation 52:83 Glycoproteomic analysis of human fibrinogen reveals novel regions of O-glycosylation. 23050552 6 67 theme ion 775:777 arg1 trap-MS/MS 779:788 ion trap-MS/MS 775:788 ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages 775:878 Glycopeptides were analyzed by ion trap-MS/MS which displayed fragmentations of glycosidic linkages and some peptide backbone cleavages. 23050552 9 68 theme novel 1145:1149 arg1 all 1196:1198 all 1196:1198 all 1196:1198 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 9 68 theme novel 1145:1149 arg1 regions 1167:1173 seven novel O-glycosylation regions 1139:1173 seven novel O-glycosylation regions in human fibrinogen 1139:1193 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 9 69 from regions 1167:1173 arg1 fibrinogen 1184:1193 human fibrinogen 1178:1193 human fibrinogen 1178:1193 Moreover, we describe seven novel O-glycosylation regions in human fibrinogen, all occupied by a monosialylated T-antigen. 23050552 5 70 theme trap 719:722 arg1 -MS 728:730 nanoLC-ESI-ion trap (IT)-MS 704:730 nanoLC-ESI-ion trap (IT)-MS 704:730 Here, we make use of tryptic- and proteinase K-generated fibrinogen glycopeptides for the detailed analysis of the protein's O-glycosylation by combining information obtained from both one- and two-dimensional nanoLC-ESI-ion trap (IT)-MS approaches. 23050552 3 71 gly N-glycosylation 358:372 arg2 one 354:356 one 354:356 one 354:356 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 3 71 gly N-glycosylation 358:372 arg2 site 374:377 one N-glycosylation site 354:377 one N-glycosylation site 354:377 The beta and gamma subunits each contain one N-glycosylation site, each of which is occupied by a biantennary N-glycan. 23050552 7 72 theme peptide 950:956 arg1 backbone 958:965 the peptide backbone 946:965 the peptide backbone 946:965 MS3 spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 21676880 0 0 theme membrane 92:99 arg1 biogenesis 109:118 membrane protein biogenesis 92:118 membrane protein biogenesis 92:118 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 6 1 theme new 1331:1333 arg1 mechanism 1344:1352 a new biogenic mechanism 1329:1352 a new biogenic mechanism for human disease 1329:1370 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 5 2 theme co-translational 1126:1141 arg1 site 1143:1146 a co-translational site 1124:1146 a co-translational site 1124:1146 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 2 3 gly N-glycosylation 421:435 arg1 site 454:457 one consensus site 440:457 one consensus site 440:457 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 4 4 theme cell 922:925 arg1 surface 927:933 the cell surface 918:933 the cell surface 918:933 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 5 5 theme range 978:982 arg1 inhibition 984:993 This long range inhibition 968:993 This long range inhibition 968:993 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 5 6 theme long 973:976 arg1 inhibition 984:993 This long range inhibition 968:993 This long range inhibition 968:993 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 0 7 theme protein 101:107 arg1 biogenesis 109:118 membrane protein biogenesis 92:118 membrane protein biogenesis 92:118 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 6 8 theme point 1253:1257 arg1 mutation 1259:1266 a single point mutation 1244:1266 a single point mutation 1244:1266 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 4 9 theme co-translational 768:783 arg1 site 785:788 the co-translational site 764:788 the co-translational site 764:788 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 4 10 theme post-translational 832:849 arg1 site 851:854 the post-translational site 828:854 the post-translational site 828:854 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 1 11 theme membrane 152:159 arg1 proteins 161:168 membrane proteins 152:168 membrane proteins 152:168 N-Glycosylation of membrane proteins is critical for their proper folding, co-assembly and subsequent matriculation through the secretory pathway. 21676880 2 12 theme congenital 508:517 arg1 deafness 519:526 congenital deafness 508:526 congenital deafness 508:526 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 3 13 gly N-glycosylation 565:579 arg2 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 13 gly N-glycosylation 565:579 arg2 two 552:554 two 552:554 two 552:554 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 13 gly N-glycosylation 565:579 arg2 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 13 gly N-glycosylation 565:579 arg2 site 636:639 a consensus site 624:639 a consensus site 624:639 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 4 14 theme KCNE1 885:889 arg1 subunits 891:898 unglycosylated KCNE1 subunits 870:898 unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel 870:965 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 3 15 dep site 615:618 arg1 residues 645:652 ∼20 residues 641:652 ∼20 residues 641:652 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 6 16 theme single 1246:1251 arg1 mutation 1259:1266 a single point mutation 1244:1266 a single point mutation 1244:1266 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 2 17 theme point 392:396 arg1 mutations 398:406 point mutations 392:406 point mutations that prevent N-glycosylation at one consensus site 392:457 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 3 18 dep acquires 677:684 arg1 post-translational 721:738 post-translational 721:738 post-translational 721:738 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 4 19 gly glycosylation 811:823 arg1 site 851:854 the post-translational site 828:854 the post-translational site 828:854 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 2 20 theme consensus 444:452 arg1 site 454:457 one consensus site 440:457 one consensus site 440:457 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 1 21 theme proper 192:197 arg1 folding 199:205 their proper folding 186:205 their proper folding 186:205 N-Glycosylation of membrane proteins is critical for their proper folding, co-assembly and subsequent matriculation through the secretory pathway. 21676880 5 22 theme mutagenic 1061:1069 arg1 conversion 1071:1080 mutagenic conversion 1061:1080 mutagenic conversion of the KCNE1 post-translational site into a co-translational site 1061:1146 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 5 23 theme post-translational 1018:1035 arg1 N-glycosylation 1037:1051 post-translational N-glycosylation 1018:1051 post-translational N-glycosylation 1018:1051 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 2 24 theme type 334:337 arg1 K 361:361 type I transmembrane KCNE1 K 334:361 type I transmembrane KCNE1 K 334:361 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 0 25 theme Post-translational 0:17 arg1 N-glycosylation 19:33 Post-translational N-glycosylation 0:33 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 0:131 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 6 26 theme biogenic 1335:1342 arg1 mechanism 1344:1352 a new biogenic mechanism 1329:1352 a new biogenic mechanism for human disease 1329:1370 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 3 27 theme N-glycosylation 565:579 arg1 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 27 theme N-glycosylation 565:579 arg1 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 27 theme N-glycosylation 565:579 arg1 site 636:639 a consensus site 624:639 a consensus site 624:639 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 6 28 theme human 1358:1362 arg1 disease 1364:1370 human disease 1358:1370 human disease 1358:1370 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 0 29 theme type 38:41 arg1 peptides 65:72 type I transmembrane KCNE1 peptides 38:72 type I transmembrane KCNE1 peptides 38:72 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 3 30 contain has 548:550 arg2 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 30 contain has 548:550 arg2 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 30 contain has 548:550 arg1 KCNE1 542:546 KCNE1 542:546 KCNE1 542:546 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 30 contain has 548:550 arg2 site 636:639 a consensus site 624:639 a consensus site 624:639 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 1 31 gly N-Glycosylation 133:147 arg1 proteins 161:168 membrane proteins 152:168 membrane proteins 152:168 N-Glycosylation of membrane proteins is critical for their proper folding, co-assembly and subsequent matriculation through the secretory pathway. 21676880 3 32 dep sites 581:585 arg1 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 32 dep sites 581:585 arg1 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 32 dep sites 581:585 arg1 site 636:639 a consensus site 624:639 a consensus site 624:639 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 2 33 theme addition 322:329 arg1 kinetics 301:308 the kinetics 297:308 the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness 297:526 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 4 34 gly unglycosylated 870:883 arg1 subunits 891:898 unglycosylated KCNE1 subunits 870:898 unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel 870:965 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 4 35 theme K 954:954 arg1 channel 959:965 their cognate K(+) channel 940:965 their cognate K(+) channel 940:965 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 3 36 theme ∼20 641:643 arg1 residues 645:652 ∼20 residues 641:652 ∼20 residues 641:652 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 2 37 theme N-glycan 313:320 arg1 addition 322:329 N-glycan addition 313:329 N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness 313:526 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 2 38 theme channel 366:372 arg1 β-subunits 374:383 type I transmembrane KCNE1 K(+) channel β-subunits 334:383 type I transmembrane KCNE1 K(+) channel β-subunits 334:383 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 0 39 theme transmembrane 45:57 arg1 peptides 65:72 type I transmembrane KCNE1 peptides 38:72 type I transmembrane KCNE1 peptides 38:72 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 1 40 theme subsequent 224:233 arg1 matriculation 235:247 subsequent matriculation 224:247 subsequent matriculation 224:247 N-Glycosylation of membrane proteins is critical for their proper folding, co-assembly and subsequent matriculation through the secretory pathway. 21676880 0 41 theme I 43:43 arg1 peptides 65:72 type I transmembrane KCNE1 peptides 38:72 type I transmembrane KCNE1 peptides 38:72 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 3 42 theme consensus 626:634 arg1 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 42 theme consensus 626:634 arg1 site 636:639 a consensus site 624:639 a consensus site 624:639 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 0 43 gly N-glycosylation 19:33 arg1 peptides 65:72 type I transmembrane KCNE1 peptides 38:72 type I transmembrane KCNE1 peptides 38:72 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 0 44 theme peptides 65:72 arg1 N-glycosylation 19:33 Post-translational N-glycosylation 0:33 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 0:131 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 6 45 theme multiple 1303:1310 arg1 sites 1312:1316 multiple sites 1303:1316 multiple sites 1303:1316 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 2 46 theme K 361:361 arg1 β-subunits 374:383 type I transmembrane KCNE1 K(+) channel β-subunits 334:383 type I transmembrane KCNE1 K(+) channel β-subunits 334:383 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 0 47 theme KCNE1 59:63 arg1 peptides 65:72 type I transmembrane KCNE1 peptides 38:72 type I transmembrane KCNE1 peptides 38:72 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 5 48 theme anterograde 1184:1194 arg1 trafficking 1196:1206 anterograde trafficking 1184:1206 anterograde trafficking 1184:1206 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 4 49 theme unglycosylated 870:883 arg1 subunits 891:898 unglycosylated KCNE1 subunits 870:898 unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel 870:965 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 2 50 theme KCNE1 355:359 arg1 K 361:361 type I transmembrane KCNE1 K 334:361 type I transmembrane KCNE1 K 334:361 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 3 51 theme distinct 556:563 arg1 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 51 theme distinct 556:563 arg1 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 51 theme distinct 556:563 arg1 site 636:639 a consensus site 624:639 a consensus site 624:639 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 4 52 theme cognate 946:952 arg1 K 954:954 their cognate K 940:954 their cognate K 940:954 Mutations that ablate the co-translational site concomitantly reduce glycosylation at the post-translational site, resulting in unglycosylated KCNE1 subunits that cannot reach the cell surface with their cognate K(+) channel. 21676880 5 53 theme KCNE1 1089:1093 arg1 site 1114:1117 the KCNE1 post-translational site 1085:1117 the KCNE1 post-translational site 1085:1117 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 3 54 theme typical 590:596 arg1 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 54 theme typical 590:596 arg1 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 2 55 theme transmembrane 341:353 arg1 K 361:361 type I transmembrane KCNE1 K 334:361 type I transmembrane KCNE1 K 334:361 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 3 56 theme co-translational 598:613 arg1 site 615:618 a typical co-translational site 588:618 a typical co-translational site 588:618 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 3 56 theme co-translational 598:613 arg1 sites 581:585 two distinct N-glycosylation sites 552:585 two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational) 552:739 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 5 57 theme post-translational 1095:1112 arg1 site 1114:1117 the KCNE1 post-translational site 1085:1117 the KCNE1 post-translational site 1085:1117 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 6 58 theme N-glycan 1280:1287 arg1 attachment 1289:1298 N-glycan attachment 1280:1298 N-glycan attachment 1280:1298 These results directly explain how a single point mutation can prevent N-glycan attachment at multiple sites, providing a new biogenic mechanism for human disease. 21676880 2 59 theme I 339:339 arg1 K 361:361 type I transmembrane KCNE1 K 334:361 type I transmembrane KCNE1 K 334:361 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 2 60 theme rhythm 497:502 arg1 disorders 472:480 disorders 472:480 disorders of the cardiac rhythm and congenital deafness 472:526 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 5 61 theme site 1114:1117 arg1 conversion 1071:1080 mutagenic conversion 1061:1080 mutagenic conversion of the KCNE1 post-translational site into a co-translational site 1061:1146 This long range inhibition is highly specific for post-translational N-glycosylation because mutagenic conversion of the KCNE1 post-translational site into a co-translational site restored both monoglycosylation and anterograde trafficking. 21676880 0 62 dep N-glycosylation 19:33 arg1 implications 75:86 implications 75:86 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 0:131 Post-translational N-glycosylation of type I transmembrane KCNE1 peptides: implications for membrane protein biogenesis and disease. 21676880 1 63 theme proteins 161:168 arg1 N-Glycosylation 133:147 N-Glycosylation 133:147 N-Glycosylation of membrane proteins 133:168 N-Glycosylation of membrane proteins is critical for their proper folding, co-assembly and subsequent matriculation through the secretory pathway. 21676880 1 64 theme secretory 261:269 arg1 pathway 271:277 the secretory pathway 257:277 the secretory pathway 257:277 N-Glycosylation of membrane proteins is critical for their proper folding, co-assembly and subsequent matriculation through the secretory pathway. 21676880 2 65 theme cardiac 489:495 arg1 rhythm 497:502 the cardiac rhythm 485:502 the cardiac rhythm 485:502 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 21676880 3 66 theme protein 702:708 arg1 synthesis 710:718 protein synthesis 702:718 protein synthesis 702:718 We show that KCNE1 has two distinct N-glycosylation sites: a typical co-translational site and a consensus site ∼20 residues away that unexpectedly acquires N-glycans after protein synthesis (post-translational). 21676880 2 67 theme deafness 519:526 arg1 disorders 472:480 disorders 472:480 disorders of the cardiac rhythm and congenital deafness 472:526 Here, we examine the kinetics of N-glycan addition to type I transmembrane KCNE1 K(+) channel β-subunits, where point mutations that prevent N-glycosylation at one consensus site give rise to disorders of the cardiac rhythm and congenital deafness. 19196183 0 0 theme cells 95:99 arg1 proteins 52:59 secreted proteins 43:59 secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach 43:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 9 1 theme secreted 1336:1343 arg1 proteins 1345:1352 more low-abundance and genuinely secreted proteins 1303:1352 more low-abundance and genuinely secreted proteins 1303:1352 As a consequence, more low-abundance and genuinely secreted proteins were identified. 19196183 9 1 theme secreted 1336:1343 arg1 consequence 1290:1300 a consequence 1288:1300 a consequence 1288:1300 As a consequence, more low-abundance and genuinely secreted proteins were identified. 19196183 2 2 dep sites 324:328 arg1 respect 293:299 respect 293:299 respect 293:299 However, with respect to their glycosylation sites, little attention has been paid. 19196183 4 3 used used 597:600 arg2 methods 524:530 capture methods 516:530 capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) 516:590 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 4 theme glycopeptides 501:513 arg1 enrichment 487:496 the enrichment 483:496 the enrichment of glycopeptides 483:513 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 0 5 theme carcinoma 85:93 arg1 cells 95:99 human hepatocellular carcinoma cells 64:99 human hepatocellular carcinoma cells 64:99 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 7 6 theme HA 1085:1086 arg1 159 1109:1111 159 1109:1111 159 1109:1111 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 6 theme HA 1085:1086 arg1 265 1102:1104 265 1102:1104 265 1102:1104 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 6 theme HA 1085:1086 arg1 HC 1098:1099 HA outweighs HC 1085:1099 HA outweighs HC (265 vs 159) 1085:1112 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 6 theme HA 1085:1086 arg1 glycosites 1062:1071 glycosites 1062:1071 glycosites identified 1062:1082 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 6 7 theme direct 852:857 arg1 comparison 859:868 A direct comparison 850:868 A direct comparison between HA and HC methods 850:894 A direct comparison between HA and HC methods was also investigated for the first time. 19196183 8 8 theme glycoproteomic 1260:1273 arg1 strategy 1275:1282 this glycoproteomic strategy 1255:1282 this glycoproteomic strategy 1255:1282 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 10 9 gly glycoproteins 1381:1393 arg1 glycoproteins 1381:1393 the glycoproteins 1377:1393 the glycoproteins identified 1377:1404 Among the glycoproteins identified, alpha-fetoprotein, CD44 and laminin have been reported to be implicated in HCC and its metastasis. 19196183 7 10 gly glycopeptides 976:988 arg2 glycopeptides 976:988 glycopeptides 976:988 glycopeptides 976:988 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 2 11 gly glycosylation 310:322 arg2 sites 324:328 their glycosylation sites 304:328 their glycosylation sites 304:328 However, with respect to their glycosylation sites, little attention has been paid. 19196183 0 12 from Identification 0:13 arg1 proteins 52:59 secreted proteins 43:59 secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach 43:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 5 13 theme nano-LC-ESI-MS/MS 668:684 arg1 analysis 686:693 nano-LC-ESI-MS/MS analysis 668:693 nano-LC-ESI-MS/MS analysis 668:693 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 1 14 from proteins 182:189 arg1 prevalent 169:177 prevalent 169:177 prevalent 169:177 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 5 15 gly glycosylation 710:722 arg2 sites 724:728 300 different glycosylation sites 696:728 300 different glycosylation sites within 194 unique glycoproteins 696:760 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 5 15 gly glycosylation 710:722 arg2 300 696:698 300 696:698 300 696:698 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 7 16 from brief 941:945 arg1 superior 997:1004 superior 997:1004 superior 997:1004 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 3 17 from analysis 383:390 arg1 proteins 429:436 secreted proteins 420:436 secreted proteins of human hepatocellular carcinoma cells 420:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 7 18 theme selectivity 960:970 arg1 terms 951:955 terms 951:955 terms of selectivity for glycopeptides 951:988 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 2 19 theme glycosylation 310:322 arg1 sites 324:328 their glycosylation sites 304:328 their glycosylation sites 304:328 However, with respect to their glycosylation sites, little attention has been paid. 19196183 7 20 dep HA 1009:1010 arg1 % 1026:1026 51.3% 1022:1026 51.3% 1022:1026 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 20 dep HA 1009:1010 arg1 % 1017:1017 92.9% 1013:1017 92.9% 1013:1017 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 0 21 theme proteomics 122:131 arg1 approach 133:140 a complementary proteomics approach 106:140 a complementary proteomics approach 106:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 0 22 from proteins 52:59 arg1 Identification 0:13 Identification 0:13 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 0:141 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 1 23 from prevalent 169:177 arg1 proteins 182:189 proteins 182:189 proteins destined for extracellular environments 182:229 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 3 24 theme human 441:445 arg1 cells 472:476 human hepatocellular carcinoma cells 441:476 human hepatocellular carcinoma cells 441:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 3 25 theme carcinoma 462:470 arg1 cells 472:476 human hepatocellular carcinoma cells 441:476 human hepatocellular carcinoma cells 441:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 0 26 theme complementary 108:120 arg1 approach 133:140 a complementary proteomics approach 106:140 a complementary proteomics approach 106:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 4 27 theme capture 516:522 arg1 methods 524:530 capture methods 516:530 capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) 516:590 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 3 28 gly N-glycosylation 395:409 arg2 sites 411:415 N-glycosylation sites 395:415 N-glycosylation sites 395:415 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 5 29 gly glycosites 787:796 arg2 glycosites 787:796 172 glycosites 783:796 172 glycosites 783:796 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 3 30 theme cells 472:476 arg1 proteins 429:436 secreted proteins 420:436 secreted proteins of human hepatocellular carcinoma cells 420:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 3 31 theme N-glycosylation 395:409 arg1 sites 411:415 N-glycosylation sites 395:415 N-glycosylation sites 395:415 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 5 32 theme methods 640:646 arg1 use 628:630 the use 624:630 the use of both methods in combination with nano-LC-ESI-MS/MS analysis 624:693 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 1 33 theme extracellular 204:216 arg1 environments 218:229 extracellular environments 204:229 extracellular environments 204:229 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 3 34 theme sites 411:415 arg1 analysis 383:390 the analysis 379:390 the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells 379:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 9 35 theme low-abundance 1308:1320 arg1 proteins 1345:1352 more low-abundance and genuinely secreted proteins 1303:1352 more low-abundance and genuinely secreted proteins 1303:1352 As a consequence, more low-abundance and genuinely secreted proteins were identified. 19196183 9 35 theme low-abundance 1308:1320 arg1 consequence 1290:1300 a consequence 1288:1300 a consequence 1288:1300 As a consequence, more low-abundance and genuinely secreted proteins were identified. 19196183 5 36 gly glycoproteins 748:760 arg1 glycoproteins 748:760 194 unique glycoproteins 737:760 194 unique glycoproteins 737:760 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 3 37 theme hepatocellular 447:460 arg1 cells 472:476 human hepatocellular carcinoma cells 441:476 human hepatocellular carcinoma cells 441:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 0 38 theme sites 34:38 arg1 Identification 0:13 Identification 0:13 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 0:141 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 4 39 theme hydrazide 567:575 arg1 chemistry 577:585 hydrazide chemistry 567:585 hydrazide chemistry (HC) 567:590 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 39 theme hydrazide 567:575 arg1 HC 588:589 HC 588:589 HC 588:589 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 1 40 gly glycosylated 265:276 arg1 proteins 252:259 all secreted proteins 239:259 all secreted proteins 239:259 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 1 40 gly glycosylated 265:276 arg1 glycosylation 152:164 N-linked glycosylation 143:164 N-linked glycosylation 143:164 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 2 41 theme little 331:336 arg1 attention 338:346 little attention 331:346 little attention 331:346 However, with respect to their glycosylation sites, little attention has been paid. 19196183 4 42 gly glycopeptides 501:513 arg2 glycopeptides 501:513 glycopeptides 501:513 glycopeptides 501:513 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 0 43 theme N-glycosylation 18:32 arg1 sites 34:38 N-glycosylation sites 18:38 N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach 18:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 8 44 gly N-glycosylated 1206:1219 arg1 albumin 1184:1190 bovine serum albumin 1171:1190 bovine serum albumin 1171:1190 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 8 44 gly N-glycosylated 1206:1219 arg1 contaminants 1140:1151 unavoidable contaminants 1128:1151 unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated 1128:1219 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 8 44 gly N-glycosylated 1206:1219 arg1 actin 1161:1165 actin 1161:1165 actin 1161:1165 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 6 45 theme first 926:930 arg1 time 932:935 the first time 922:935 the first time 922:935 A direct comparison between HA and HC methods was also investigated for the first time. 19196183 1 46 dep prevalent 169:177 arg1 glycosylated 265:276 glycosylated 265:276 are glycosylated 261:276 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 0 47 theme secreted 43:50 arg1 proteins 52:59 secreted proteins 43:59 secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach 43:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 6 48 dep HA 878:879 arg1 methods 888:894 methods 888:894 methods 888:894 A direct comparison between HA and HC methods was also investigated for the first time. 19196183 8 49 theme serum 1178:1182 arg1 albumin 1184:1190 bovine serum albumin 1171:1190 bovine serum albumin 1171:1190 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 5 50 with combination 651:661 arg1 analysis 686:693 nano-LC-ESI-MS/MS analysis 668:693 nano-LC-ESI-MS/MS analysis 668:693 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 5 51 theme unique 741:746 arg1 glycoproteins 748:760 194 unique glycoproteins 737:760 194 unique glycoproteins 737:760 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 1 52 theme secreted 243:250 arg1 proteins 252:259 all secreted proteins 239:259 all secreted proteins 239:259 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 7 53 theme outweighs 1088:1096 arg1 159 1109:1111 159 1109:1111 159 1109:1111 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 53 theme outweighs 1088:1096 arg1 265 1102:1104 265 1102:1104 265 1102:1104 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 53 theme outweighs 1088:1096 arg1 HC 1098:1099 HA outweighs HC 1085:1099 HA outweighs HC (265 vs 159) 1085:1112 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 53 theme outweighs 1088:1096 arg1 glycosites 1062:1071 glycosites 1062:1071 glycosites identified 1062:1082 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 1 54 link N-linked 143:150 arg1 glycosylation 152:164 N-linked glycosylation 143:164 N-linked glycosylation 143:164 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 0 55 gly N-glycosylation 18:32 arg2 sites 34:38 N-glycosylation sites 18:38 N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach 18:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 3 56 theme secreted 420:427 arg1 proteins 429:436 secreted proteins 420:436 secreted proteins of human hepatocellular carcinoma cells 420:476 Here, we report the analysis of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells. 19196183 8 57 theme bovine 1171:1176 arg1 albumin 1184:1190 bovine serum albumin 1171:1190 bovine serum albumin 1171:1190 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 7 58 theme glycosites 1062:1071 arg1 number 1052:1057 the number 1048:1057 the number of glycosites identified, HA outweighs HC (265 vs 159) 1048:1112 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 5 59 theme different 700:708 arg1 sites 724:728 300 different glycosylation sites 696:728 300 different glycosylation sites within 194 unique glycoproteins 696:760 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 1 60 theme N-linked 143:150 arg1 glycosylation 152:164 N-linked glycosylation 143:164 N-linked glycosylation 143:164 N-linked glycosylation is prevalent in proteins destined for extracellular environments; nearly all secreted proteins are glycosylated. 19196183 5 61 theme glycosylation 710:722 arg1 sites 724:728 300 different glycosylation sites 696:728 300 different glycosylation sites within 194 unique glycoproteins 696:760 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 19196183 4 62 theme hydrophilic 537:547 arg1 HA 559:560 HA 559:560 HA 559:560 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 62 theme hydrophilic 537:547 arg1 affinity 549:556 hydrophilic affinity 537:556 hydrophilic affinity (HA) 537:561 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 63 with methods 524:530 arg1 chemistry 577:585 hydrazide chemistry 567:585 hydrazide chemistry (HC) 567:590 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 63 with methods 524:530 arg1 HC 588:589 HC 588:589 HC 588:589 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 63 with methods 524:530 arg1 HA 559:560 HA 559:560 HA 559:560 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 4 63 with methods 524:530 arg1 affinity 549:556 hydrophilic affinity 537:556 hydrophilic affinity (HA) 537:561 For the enrichment of glycopeptides, capture methods with hydrophilic affinity (HA) and hydrazide chemistry (HC) were used complementarily. 19196183 7 64 from terms 951:955 arg1 superior 997:1004 superior 997:1004 superior 997:1004 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 0 65 theme hepatocellular 70:83 arg1 cells 95:99 human hepatocellular carcinoma cells 64:99 human hepatocellular carcinoma cells 64:99 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 7 66 gly glycosites 1062:1071 arg2 glycosites 1062:1071 glycosites 1062:1071 glycosites identified 1062:1082 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 66 gly glycosites 1062:1071 arg2 HC 1098:1099 HA outweighs HC 1085:1099 HA outweighs HC (265 vs 159) 1085:1112 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 0 67 with proteins 52:59 arg1 approach 133:140 a complementary proteomics approach 106:140 a complementary proteomics approach 106:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 7 68 from superior 997:1004 arg1 terms 951:955 terms 951:955 terms of selectivity for glycopeptides 951:988 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 7 68 from superior 997:1004 arg1 brief 941:945 brief 941:945 brief 941:945 In brief, in terms of selectivity for glycopeptides, HC is superior to HA (92.9% vs 51.3%); however, based on the number of glycosites identified, HA outweighs HC (265 vs 159). 19196183 0 69 theme human 64:68 arg1 cells 95:99 human hepatocellular carcinoma cells 64:99 human hepatocellular carcinoma cells 64:99 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 0 70 from sites 34:38 arg1 proteins 52:59 secreted proteins 43:59 secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach 43:140 Identification of N-glycosylation sites on secreted proteins of human hepatocellular carcinoma cells with a complementary proteomics approach. 19196183 8 71 theme unavoidable 1128:1138 arg1 albumin 1184:1190 bovine serum albumin 1171:1190 bovine serum albumin 1171:1190 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 8 71 theme unavoidable 1128:1138 arg1 contaminants 1140:1151 unavoidable contaminants 1128:1151 unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated 1128:1219 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 8 71 theme unavoidable 1128:1138 arg1 actin 1161:1165 actin 1161:1165 actin 1161:1165 Furthermore, unavoidable contaminants such as actin and bovine serum albumin which are not N-glycosylated could be easily depleted by using this glycoproteomic strategy. 19196183 5 72 from use 628:630 arg1 combination 651:661 combination 651:661 combination with nano-LC-ESI-MS/MS analysis 651:693 With the use of both methods in combination with nano-LC-ESI-MS/MS analysis, 300 different glycosylation sites within 194 unique glycoproteins were identified, and 172 glycosites have not been determined experimentally previously. 20511397 1 0 theme lipoprotein 327:337 arg1 clearance 339:347 remnant lipoprotein clearance 319:347 remnant lipoprotein clearance 319:347 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 5 1 theme secreted 1067:1074 arg1 apoE 1076:1079 both cellular and secreted apoE 1049:1079 both cellular and secreted apoE 1049:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 4 2 theme gel 869:871 arg1 electrophoresis 873:887 one- and two-dimensional gel electrophoresis 844:887 one- and two-dimensional gel electrophoresis 844:887 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 7 3 theme plasma 1401:1406 arg1 apoE 1408:1411 plasma apoE 1401:1411 plasma apoE 1401:1411 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 1 4 theme various 236:242 arg1 cells 244:248 various cells 236:248 various cells including hepatocytes and macrophages 236:286 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 4 theme various 236:242 arg1 macrophages 276:286 macrophages 276:286 macrophages 276:286 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 4 theme various 236:242 arg1 hepatocytes 260:270 hepatocytes 260:270 hepatocytes 260:270 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 7 5 link cell-derived 1347:1358 arg1 apoE 1360:1363 cell-derived apoE 1347:1363 cell-derived apoE 1347:1363 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 8 6 theme terminus 1444:1451 arg1 importance 1424:1433 the importance 1420:1433 the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding 1420:1519 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 1 7 gly glycoprotein 209:220 arg1 E 188:188 Apolipoprotein E 173:188 Apolipoprotein E (apoE) 173:195 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 7 gly glycoprotein 209:220 arg1 glycoprotein 209:220 a 34-kDa glycoprotein 200:220 a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages 200:286 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 5 8 dep -Hex 976:979 arg1 2 974:974 2 974:974 2 974:974 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 8 9 theme C 1442:1442 arg1 terminus 1444:1451 the C terminus 1438:1451 the C terminus of apoE 1438:1459 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 4 10 gly sialylation 768:778 arg1 apoE 805:808 cellular and secreted apoE 783:808 cellular and secreted apoE from primary human macrophages 783:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 4 11 theme human 823:827 arg1 macrophages 829:839 primary human macrophages 815:839 primary human macrophages 815:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 7 12 theme apoE 1307:1310 arg1 Comparison 1264:1273 Comparison 1264:1273 Comparison of plasma and cellular/secreted apoE from the same donor 1264:1330 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 2 13 gly glycosylated/sialylated 523:545 arg1 apoE 507:510 plasma apoE 500:510 plasma apoE 500:510 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 7 14 from Comparison 1264:1273 arg1 donor 1326:1330 the same donor 1317:1330 the same donor 1317:1330 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 0 15 theme mass 100:103 arg1 spectrometry 105:116 mass spectrometry 100:116 mass spectrometry 100:116 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 1 16 theme immune 350:355 arg1 responses 357:365 immune responses 350:365 immune responses 350:365 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 5 17 with glycoforms 949:958 arg1 2 992:992 2 992:992 2 992:992 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 5 17 with glycoforms 949:958 arg1 -Hex 976:979 (HexNAc)(2)-Hex(2)- 965:983 (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE 965:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 5 17 with glycoforms 949:958 arg1 HexNAc 966:971 HexNAc 966:971 HexNAc 966:971 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 5 17 with glycoforms 949:958 arg1 NeuAc 985:989 NeuAc 985:989 NeuAc 985:989 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 8 18 contain have 1540:1543 arg1 results 1528:1534 these results 1522:1534 these results 1522:1534 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 8 18 contain have 1540:1543 arg2 implications 1555:1566 important implications 1545:1566 important implications for our understanding of apoE biochemistry 1545:1609 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 4 19 theme secreted 796:803 arg1 apoE 805:808 cellular and secreted apoE 783:808 cellular and secreted apoE from primary human macrophages 783:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 3 20 theme plasma 604:609 arg1 apoE 611:614 plasma apoE 604:614 plasma apoE 604:614 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 5 21 theme cellular 1054:1061 arg1 apoE 1076:1079 both cellular and secreted apoE 1049:1079 both cellular and secreted apoE 1049:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 4 22 theme mass 893:896 arg1 spectrometry 898:909 mass spectrometry 893:909 mass spectrometry 893:909 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 8 23 theme biochemistry 1598:1609 arg1 understanding 1576:1588 our understanding 1572:1588 our understanding of apoE biochemistry 1572:1609 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 4 24 theme primary 815:821 arg1 macrophages 829:839 primary human macrophages 815:839 primary human macrophages 815:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 2 25 link cell-derived 552:563 arg1 apoE 565:568 cell-derived apoE 552:568 cell-derived apoE 552:568 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 2 26 theme multiple 447:454 arg1 glycoforms 484:493 multiple glycosylated and sialylated glycoforms 447:493 multiple glycosylated and sialylated glycoforms 447:493 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 2 27 theme sialylated 473:482 arg1 glycoforms 484:493 multiple glycosylated and sialylated glycoforms 447:493 multiple glycosylated and sialylated glycoforms 447:493 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 8 28 theme apoE 1456:1459 arg1 terminus 1444:1451 the C terminus 1438:1451 the C terminus of apoE 1438:1459 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 1 29 theme Apolipoprotein 173:186 arg1 E 188:188 Apolipoprotein E 173:188 Apolipoprotein E (apoE) 173:195 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 29 theme Apolipoprotein 173:186 arg1 glycoprotein 209:220 a 34-kDa glycoprotein 200:220 a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages 200:286 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 29 theme Apolipoprotein 173:186 arg1 apoE 191:194 apoE 191:194 apoE 191:194 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 30 theme Alzheimer 368:376 arg1 disease 378:384 Alzheimer disease 368:384 Alzheimer disease 368:384 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 3 31 theme glycan 583:588 arg1 structures 590:599 the glycan structures 579:599 the glycan structures on plasma apoE 579:614 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 7 32 gly sialylated 1385:1394 arg1 apoE 1360:1363 cell-derived apoE 1347:1363 cell-derived apoE 1347:1363 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 6 33 theme attachment 1252:1261 arg1 site 1237:1240 a novel site 1229:1240 a novel site of glycan attachment 1229:1261 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 6 33 theme attachment 1252:1261 arg1 attachment 1252:1261 glycan attachment 1245:1261 glycan attachment 1245:1261 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 6 33 theme attachment 1252:1261 arg1 Ser 1217:1219 Ser 1217:1219 Ser(290) 1217:1224 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 7 34 theme cell-derived 1347:1358 arg1 apoE 1360:1363 cell-derived apoE 1347:1363 cell-derived apoE 1347:1363 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 8 35 theme important 1545:1553 arg1 implications 1555:1566 important implications 1545:1566 important implications for our understanding of apoE biochemistry 1545:1609 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 8 36 theme apoE 1475:1478 arg1 solubility 1480:1489 apoE solubility 1475:1489 apoE solubility 1475:1489 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 0 37 link macrophage-derived 33:50 arg1 E 73:73 macrophage-derived human apolipoprotein E 33:73 macrophage-derived human apolipoprotein E 33:73 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 4 38 theme two-dimensional 853:867 arg1 electrophoresis 873:887 one- and two-dimensional gel electrophoresis 844:887 one- and two-dimensional gel electrophoresis 844:887 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 5 39 located detected 1025:1032 arg2 glycan 1018:1023 the most complex glycan 1001:1023 the most complex glycan detected on Thr(194) in both cellular and secreted apoE 1001:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 5 39 located detected 1025:1032 arg1 Thr 1037:1039 Thr 1037:1039 Thr(194) in both cellular and secreted apoE 1037:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 5 39 located detected 1025:1032 arg1 194 1041:1043 194 1041:1043 194 1041:1043 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 2 40 theme plasma 426:431 arg1 apoE 433:436 Cellular apoE and plasma apoE 408:436 apoE 433:436 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 3 41 theme complicated 653:663 arg1 structures 665:674 the more complicated structures 644:674 the more complicated structures on plasma and cellular/secreted apoE 644:711 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 4 42 theme cellular 783:790 arg1 apoE 805:808 cellular and secreted apoE 783:808 cellular and secreted apoE from primary human macrophages 783:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 2 43 theme glycosylated 456:467 arg1 glycoforms 484:493 multiple glycosylated and sialylated glycoforms 447:493 multiple glycosylated and sialylated glycoforms 447:493 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 7 44 theme cellular/secreted 1289:1305 arg1 apoE 1307:1310 cellular/secreted apoE 1289:1310 cellular/secreted apoE 1289:1310 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 0 45 theme human 52:56 arg1 E 73:73 macrophage-derived human apolipoprotein E 33:73 macrophage-derived human apolipoprotein E 33:73 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 8 46 theme lipid 1507:1511 arg1 binding 1513:1519 lipid binding 1507:1519 lipid binding 1507:1519 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 4 47 gly glycosylation 750:762 arg1 apoE 805:808 cellular and secreted apoE 783:808 cellular and secreted apoE from primary human macrophages 783:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 0 48 gly Glycosylation 0:12 arg1 E 73:73 macrophage-derived human apolipoprotein E 33:73 macrophage-derived human apolipoprotein E 33:73 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 2 49 theme Cellular 408:415 arg1 apoE 417:420 Cellular apoE and plasma apoE 408:436 apoE 417:420 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 0 50 theme macrophage-derived 33:50 arg1 E 73:73 macrophage-derived human apolipoprotein E 33:73 macrophage-derived human apolipoprotein E 33:73 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 0 51 theme novel 134:138 arg1 glycosylation 148:160 glycosylation 148:160 glycosylation 148:160 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 0 51 theme novel 134:138 arg1 site 140:143 a novel site 132:143 a novel site of glycosylation on Ser290 132:170 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 2 52 gly sialylated 473:482 arg1 glycoforms 484:493 multiple glycosylated and sialylated glycoforms 447:493 multiple glycosylated and sialylated glycoforms 447:493 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 2 53 theme cell-derived 552:563 arg1 apoE 565:568 cell-derived apoE 552:568 cell-derived apoE 552:568 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 3 54 from structures 665:674 arg1 plasma 679:684 plasma 679:684 plasma 679:684 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 3 54 from structures 665:674 arg1 apoE 708:711 cellular/secreted apoE 690:711 cellular/secreted apoE 690:711 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 0 55 theme E 73:73 arg1 sialylation 18:28 sialylation 18:28 sialylation 18:28 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 0 55 theme E 73:73 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation 0:12 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 1 56 theme important 301:309 arg1 role 311:314 an important role 298:314 an important role 298:314 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 0 57 theme glycosylation 148:160 arg1 glycosylation 148:160 glycosylation 148:160 glycosylation 148:160 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 0 57 theme glycosylation 148:160 arg1 site 140:143 a novel site 132:143 a novel site of glycosylation on Ser290 132:170 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 5 58 from Thr 1037:1039 arg1 apoE 1076:1079 both cellular and secreted apoE 1049:1079 both cellular and secreted apoE 1049:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 0 59 gly sialylation 18:28 arg1 E 73:73 macrophage-derived human apolipoprotein E 33:73 macrophage-derived human apolipoprotein E 33:73 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 6 60 theme additional 1087:1096 arg1 glycans 1098:1104 Four additional glycans 1082:1104 Four additional glycans 1082:1104 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 0 61 theme apolipoprotein 58:71 arg1 E 73:73 macrophage-derived human apolipoprotein E 33:73 macrophage-derived human apolipoprotein E 33:73 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 5 62 theme complex 1010:1016 arg1 glycan 1018:1023 the most complex glycan 1001:1023 the most complex glycan detected on Thr(194) in both cellular and secreted apoE 1001:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 2 63 gly glycosylated 456:467 arg1 glycoforms 484:493 multiple glycosylated and sialylated glycoforms 447:493 multiple glycosylated and sialylated glycoforms 447:493 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 7 64 from donor 1326:1330 arg1 plasma 1278:1283 plasma 1278:1283 plasma 1278:1283 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 7 64 from donor 1326:1330 arg1 apoE 1307:1310 cellular/secreted apoE 1289:1310 cellular/secreted apoE 1289:1310 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 7 64 from donor 1326:1330 arg1 Comparison 1264:1273 Comparison 1264:1273 Comparison of plasma and cellular/secreted apoE from the same donor 1264:1330 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 4 65 from sialylation 768:778 arg1 macrophages 829:839 primary human macrophages 815:839 primary human macrophages 815:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 6 66 theme novel 1231:1235 arg1 site 1237:1240 a novel site 1229:1240 a novel site of glycan attachment 1229:1261 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 6 66 theme novel 1231:1235 arg1 attachment 1252:1261 glycan attachment 1245:1261 glycan attachment 1245:1261 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 6 66 theme novel 1231:1235 arg1 Ser 1217:1219 Ser 1217:1219 Ser(290) 1217:1224 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 3 67 theme cellular/secreted 690:706 arg1 apoE 708:711 cellular/secreted apoE 690:711 cellular/secreted apoE 690:711 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 7 68 theme same 1321:1324 arg1 donor 1326:1330 the same donor 1317:1330 the same donor 1317:1330 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 6 69 theme glycan 1245:1250 arg1 attachment 1252:1261 glycan attachment 1245:1261 glycan attachment 1245:1261 Four additional glycans were identified on apoE(283-299), and using beta-elimination/alkylation by methylamine in vitro, we identified Ser(290) as a novel site of glycan attachment. 20511397 4 70 from glycosylation 750:762 arg1 macrophages 829:839 primary human macrophages 815:839 primary human macrophages 815:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 0 71 from site 140:143 arg1 Ser290 165:170 Ser290 165:170 Ser290 165:170 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 4 72 theme apoE 805:808 arg1 sialylation 768:778 sialylation 768:778 sialylation 768:778 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 4 72 theme apoE 805:808 arg1 glycosylation 750:762 glycosylation 750:762 glycosylation 750:762 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 0 73 gly glycosylation 148:160 arg2 glycosylation 148:160 glycosylation 148:160 glycosylation 148:160 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 0 73 gly glycosylation 148:160 arg2 site 140:143 a novel site 132:143 a novel site of glycosylation on Ser290 132:170 Glycosylation and sialylation of macrophage-derived human apolipoprotein E analyzed by SDS-PAGE and mass spectrometry: evidence for a novel site of glycosylation on Ser290. 20511397 8 74 theme apoE 1593:1596 arg1 biochemistry 1598:1609 apoE biochemistry 1593:1609 apoE biochemistry 1593:1609 Given the importance of the C terminus of apoE in regulating apoE solubility, stability, and lipid binding, these results may have important implications for our understanding of apoE biochemistry. 20511397 4 75 theme one- 844:847 arg1 electrophoresis 873:887 one- and two-dimensional gel electrophoresis 844:887 one- and two-dimensional gel electrophoresis 844:887 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 5 76 theme different 939:947 arg1 glycoforms 949:958 eight different glycoforms 933:958 eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE 933:1079 Our results identify eight different glycoforms with (HexNAc)(2)-Hex(2)-(NeuAc)(2) being the most complex glycan detected on Thr(194) in both cellular and secreted apoE. 20511397 3 77 from structures 590:599 arg1 apoE 611:614 plasma apoE 604:614 plasma apoE 604:614 Some of the glycan structures on plasma apoE are characterized; however, the more complicated structures on plasma and cellular/secreted apoE remain unidentified. 20511397 2 78 theme plasma 500:505 arg1 apoE 507:510 plasma apoE 500:510 plasma apoE 500:510 Cellular apoE and plasma apoE exist as multiple glycosylated and sialylated glycoforms with plasma apoE being less glycosylated/sialylated than cell-derived apoE. 20511397 1 79 theme remnant 319:325 arg1 clearance 339:347 remnant lipoprotein clearance 319:347 remnant lipoprotein clearance 319:347 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 7 80 theme plasma 1278:1283 arg1 Comparison 1264:1273 Comparison 1264:1273 Comparison of plasma and cellular/secreted apoE from the same donor 1264:1330 Comparison of plasma and cellular/secreted apoE from the same donor confirmed that cell-derived apoE is more extensively sialylated than plasma apoE. 20511397 4 81 from macrophages 829:839 arg1 apoE 805:808 cellular and secreted apoE 783:808 cellular and secreted apoE from primary human macrophages 783:839 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 4 81 from macrophages 829:839 arg1 sialylation 768:778 sialylation 768:778 sialylation 768:778 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 4 81 from macrophages 829:839 arg1 glycosylation 750:762 glycosylation 750:762 glycosylation 750:762 We investigated glycosylation and sialylation of cellular and secreted apoE from primary human macrophages by one- and two-dimensional gel electrophoresis and mass spectrometry. 20511397 1 82 theme 34-kDa 202:207 arg1 E 188:188 Apolipoprotein E 173:188 Apolipoprotein E (apoE) 173:195 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20511397 1 82 theme 34-kDa 202:207 arg1 glycoprotein 209:220 a 34-kDa glycoprotein 200:220 a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages 200:286 Apolipoprotein E (apoE) is a 34-kDa glycoprotein secreted from various cells including hepatocytes and macrophages and plays an important role in remnant lipoprotein clearance, immune responses, Alzheimer disease, and atherosclerosis. 20006580 3 0 from oocytes 441:447 arg1 recordings 417:426 Two-electrode voltage-clamp recordings 389:426 Two-electrode voltage-clamp recordings from Xenopus oocytes 389:447 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 1 1 theme subunits 151:158 arg1 loop 121:124 the first external loop 102:124 the first external loop of mouse and human TRESK subunits 102:158 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 0 2 theme TRESK 88:92 arg1 expression 47:56 cell surface expression 34:56 cell surface expression of two-pore-domain K+ channel TRESK 34:92 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 3 3 theme wildtype 544:551 arg1 TRESK 553:557 wildtype TRESK 544:557 wildtype TRESK 544:557 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 3 4 theme Xenopus 433:439 arg1 oocytes 441:447 Xenopus oocytes 433:447 Xenopus oocytes 433:447 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 1 5 gly N-glycosylation 171:185 arg2 one 160:162 one 160:162 one 160:162 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 1 5 gly N-glycosylation 171:185 arg2 sites 197:201 one or two N-glycosylation consensus sites 160:201 one or two N-glycosylation consensus sites 160:201 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 1 5 gly N-glycosylation 171:185 arg2 two 167:169 two 167:169 two 167:169 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 2 6 theme heterologous 364:375 arg1 expression 377:386 heterologous expression 364:386 heterologous expression 364:386 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 0 7 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 3 8 theme current 463:469 arg1 amplitudes 471:480 current amplitudes 463:480 current amplitudes of N-glycosylation mutants 463:507 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 3 9 theme N-glycosylation 485:499 arg1 mutants 501:507 N-glycosylation mutants 485:507 N-glycosylation mutants 485:507 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 1 10 theme N-glycosylation 171:185 arg1 sites 197:201 one or two N-glycosylation consensus sites 160:201 one or two N-glycosylation consensus sites 160:201 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 0 11 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 4 12 theme GFP-tagged 595:604 arg1 subunits 612:619 GFP-tagged TRESK subunits 595:619 GFP-tagged TRESK subunits 595:619 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 2 13 theme orthologues 318:328 arg1 residue 302:308 a single residue 293:308 a single residue of both orthologues 293:328 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 2 13 theme orthologues 318:328 arg1 orthologues 318:328 both orthologues 313:328 both orthologues 313:328 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 4 14 from surface 694:700 arg1 intensity 672:680 fluorescence intensity 659:680 fluorescence intensity at the cell surface 659:700 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 1 15 theme consensus 187:195 arg1 sites 197:201 one or two N-glycosylation consensus sites 160:201 one or two N-glycosylation consensus sites 160:201 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 0 16 theme cell 34:37 arg1 expression 47:56 cell surface expression 34:56 cell surface expression of two-pore-domain K+ channel TRESK 34:92 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 1 17 theme first 106:110 arg1 loop 121:124 the first external loop 102:124 the first external loop of mouse and human TRESK subunits 102:158 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 2 18 dep found 334:338 arg1 Using 234:238 Using 234:238 Using site directed mutagenesis and Western immunoblotting 234:291 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 5 19 theme N-glycosylation 754:768 arg1 mutants 770:776 the N-glycosylation mutants 750:776 the N-glycosylation mutants 750:776 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 1 20 theme external 112:119 arg1 loop 121:124 the first external loop 102:124 the first external loop of mouse and human TRESK subunits 102:158 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 2 21 theme single 295:300 arg1 residue 302:308 a single residue 293:308 a single residue of both orthologues 293:328 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 2 21 theme single 295:300 arg1 orthologues 318:328 both orthologues 313:328 both orthologues 313:328 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 3 22 theme voltage-clamp 403:415 arg1 recordings 417:426 Two-electrode voltage-clamp recordings 389:426 Two-electrode voltage-clamp recordings from Xenopus oocytes 389:447 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 5 23 theme lower 822:826 arg1 amplitudes 836:845 their lower current amplitudes 816:845 their lower current amplitudes 816:845 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 4 24 theme TRESK 606:610 arg1 subunits 612:619 GFP-tagged TRESK subunits 595:619 GFP-tagged TRESK subunits 595:619 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 5 25 theme mutants 770:776 arg1 Signals 739:745 Signals 739:745 Signals of the N-glycosylation mutants 739:776 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 2 26 theme site 240:243 arg1 mutagenesis 254:264 site directed mutagenesis 240:264 site directed mutagenesis 240:264 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 3 27 theme mutants 501:507 arg1 amplitudes 471:480 current amplitudes 463:480 current amplitudes of N-glycosylation mutants 463:507 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 0 28 theme surface 39:45 arg1 expression 47:56 cell surface expression 34:56 cell surface expression of two-pore-domain K+ channel TRESK 34:92 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 5 29 theme current 828:834 arg1 amplitudes 836:845 their lower current amplitudes 816:845 their lower current amplitudes 816:845 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 3 30 theme Two-electrode 389:401 arg1 recordings 417:426 Two-electrode voltage-clamp recordings 389:426 Two-electrode voltage-clamp recordings from Xenopus oocytes 389:447 Two-electrode voltage-clamp recordings from Xenopus oocytes revealed that current amplitudes of N-glycosylation mutants were reduced by 80% as compared to wildtype TRESK. 20006580 4 31 theme cell 689:692 arg1 surface 694:700 the cell surface 685:700 the cell surface 685:700 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 0 32 theme two-pore-domain 61:75 arg1 TRESK 88:92 two-pore-domain K+ channel TRESK 61:92 two-pore-domain K+ channel TRESK 61:92 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 4 33 theme confocal 718:725 arg1 microscopy 727:736 confocal microscopy 718:736 confocal microscopy 718:736 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 2 34 gly glycosylated 346:357 arg1 residue 302:308 a single residue 293:308 a single residue of both orthologues 293:328 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 2 34 gly glycosylated 346:357 arg1 orthologues 318:328 both orthologues 313:328 both orthologues 313:328 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 4 35 theme Xenopus 639:645 arg1 oocytes 647:653 Xenopus oocytes 639:653 Xenopus oocytes 639:653 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 5 36 theme inadequate 873:882 arg1 expression 892:901 inadequate surface expression 873:901 inadequate surface expression of the channel 873:916 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 4 37 theme membrane 575:582 arg1 targeting 584:592 membrane targeting 575:592 membrane targeting 575:592 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 20006580 2 38 theme Western 270:276 arg1 immunoblotting 278:291 Western immunoblotting 270:291 Western immunoblotting 270:291 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 1 39 theme mouse 129:133 arg1 subunits 151:158 mouse and human TRESK subunits 129:158 mouse and human TRESK subunits 129:158 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 5 40 theme surface 884:890 arg1 expression 892:901 inadequate surface expression 873:901 inadequate surface expression of the channel 873:916 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 0 41 theme channel 80:86 arg1 TRESK 88:92 two-pore-domain K+ channel TRESK 61:92 two-pore-domain K+ channel TRESK 61:92 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 5 42 theme channel 910:916 arg1 expression 892:901 inadequate surface expression 873:901 inadequate surface expression of the channel 873:916 Signals of the N-glycosylation mutants were reduced by >50%, indicating that their lower current amplitudes substantially result from inadequate surface expression of the channel. 20006580 1 43 theme human 139:143 arg1 subunits 151:158 mouse and human TRESK subunits 129:158 mouse and human TRESK subunits 129:158 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 0 44 theme K+ 77:78 arg1 TRESK 88:92 two-pore-domain K+ channel TRESK 61:92 two-pore-domain K+ channel TRESK 61:92 N-linked glycosylation determines cell surface expression of two-pore-domain K+ channel TRESK. 20006580 2 45 theme directed 245:252 arg1 mutagenesis 254:264 site directed mutagenesis 240:264 site directed mutagenesis 240:264 Using site directed mutagenesis and Western immunoblotting a single residue of both orthologues was found to be glycosylated upon heterologous expression. 20006580 1 46 theme TRESK 145:149 arg1 subunits 151:158 mouse and human TRESK subunits 129:158 mouse and human TRESK subunits 129:158 Within the first external loop of mouse and human TRESK subunits one or two N-glycosylation consensus sites were identified, respectively. 20006580 4 47 theme fluorescence 659:670 arg1 intensity 672:680 fluorescence intensity 659:680 fluorescence intensity at the cell surface 659:700 To investigate membrane targeting, GFP-tagged TRESK subunits were expressed in Xenopus oocytes and fluorescence intensity at the cell surface was measured by confocal microscopy. 15173186 8 0 theme secondary 1244:1252 arg1 contents 1264:1271 secondary structure contents 1244:1271 secondary structure contents similar to glycoprotein Ib 1244:1298 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 9 1 from located 1363:1369 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 9 1 from located 1363:1369 arg1 domains 1392:1398 the extracellular domains 1374:1398 the extracellular domains of other human Toll-like receptors 1374:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 9 2 located located 1363:1369 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 9 2 located located 1363:1369 arg1 domains 1392:1398 the extracellular domains 1374:1398 the extracellular domains of other human Toll-like receptors 1374:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 9 2 located located 1363:1369 arg2 sites 1353:1357 consensus glycosylation sites 1329:1357 consensus glycosylation sites 1329:1357 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 8 3 gly glycoprotein 1284:1295 arg1 glycoprotein 1284:1295 glycoprotein Ib 1284:1298 glycoprotein Ib 1284:1298 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 3 4 theme bacterial 537:545 arg1 lipopolysaccharide 547:564 bacterial lipopolysaccharide 537:564 bacterial lipopolysaccharide 537:564 For example, Toll-like receptor 2 appears to mediate responses to bacterial peptidoglycan and acylated lipoproteins and Toll-like receptor 4 to bacterial lipopolysaccharide. 15173186 4 5 theme structures 637:646 arg1 recognition 616:626 recognition 616:626 recognition of these structures 616:646 However, the structural principles that underlie recognition of these structures are poorly understood. 15173186 5 6 theme extracellular 726:738 arg1 domains 740:746 their extracellular domains 720:746 their extracellular domains 720:746 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 11 7 theme extracellular 1651:1663 arg1 domain 1665:1670 Toll-like receptor 2 extracellular domain 1630:1670 Toll-like receptor 2 extracellular domain 1630:1670 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 1 8 theme receptors 256:264 arg1 group 205:209 a group 203:209 a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors 203:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 1 8 theme receptors 256:264 arg1 receptors 287:295 so-called Toll-like receptors 267:295 so-called Toll-like receptors 267:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 1 8 theme receptors 256:264 arg1 receptors 256:264 evolutionarily related, germ line-encoded receptors 214:264 evolutionarily related, germ line-encoded receptors 214:264 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 9 9 gly glycosylation 1339:1351 arg2 sites 1353:1357 consensus glycosylation sites 1329:1357 consensus glycosylation sites 1329:1357 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 1 10 theme immune 168:173 arg1 defense 175:181 innate immune defense 161:181 innate immune defense 161:181 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 5 11 theme leucine-rich 696:707 arg1 repeats 709:715 leucine-rich repeats 696:715 leucine-rich repeats in their extracellular domains 696:746 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 11 12 theme predicted 1681:1689 arg1 sites 1705:1709 all four predicted glycosylation sites 1672:1709 all four predicted glycosylation sites 1672:1709 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 12 13 theme protein 1912:1918 arg1 secretion 1920:1928 efficient protein secretion 1902:1928 efficient protein secretion 1902:1928 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 9 14 theme consensus 1329:1337 arg1 sites 1353:1357 consensus glycosylation sites 1329:1357 consensus glycosylation sites 1329:1357 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 1 15 dep related 229:235 arg1 line-encoded 243:254 line-encoded 243:254 line-encoded 243:254 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 5 16 theme solenoid 779:786 arg1 structures 788:797 solenoid structures 779:797 solenoid structures 779:797 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 7 17 theme pattern 1110:1116 arg1 recognition 1118:1128 pattern recognition 1110:1128 pattern recognition 1110:1128 Toll-like receptor glycosylation is also likely to influence receptor surface representation, trafficking, and pattern recognition. 15173186 9 18 from domains 1392:1398 arg1 located 1363:1369 located 1363:1369 located 1363:1369 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 1 19 theme Most 122:125 arg1 organisms 134:142 Most higher organisms 122:142 Most higher organisms 122:142 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 6 20 contain contain 889:895 arg2 sites 930:934 N-linked glycosylation consensus sites 897:934 N-linked glycosylation consensus sites 897:934 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 6 20 contain contain 889:895 arg1 receptors 879:887 all Toll-like receptors 865:887 all Toll-like receptors 865:887 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 7 21 theme surface 1069:1075 arg1 representation 1077:1090 receptor surface representation 1060:1090 receptor surface representation 1060:1090 Toll-like receptor glycosylation is also likely to influence receptor surface representation, trafficking, and pattern recognition. 15173186 12 22 theme Toll-like 1842:1850 arg1 sites 1877:1881 Toll-like receptor 2 glycosylation sites 1842:1881 The remaining Toll-like receptor 2 glycosylation sites 1828:1881 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 12 23 gly glycosylation 1863:1875 arg2 sites 1877:1881 Toll-like receptor 2 glycosylation sites 1842:1881 The remaining Toll-like receptor 2 glycosylation sites 1828:1881 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 12 23 gly glycosylation 1863:1875 arg2 2 1861:1861 2 1861:1861 2 1861:1861 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 5 24 theme platelet 825:832 arg1 Ib 847:848 platelet glycoprotein Ib 825:848 platelet glycoprotein Ib 825:848 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 9 25 theme other 1403:1407 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 10 26 theme Toll-like 1554:1562 arg1 receptors 1564:1572 different Toll-like receptors 1544:1572 different Toll-like receptors 1544:1572 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 11 27 theme Toll-like 1630:1638 arg1 receptor 1640:1647 Toll-like receptor 2 1630:1649 Toll-like receptor 2 extracellular domain 1630:1670 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 0 28 theme efficient 84:92 arg1 biosynthesis 94:105 efficient biosynthesis 84:105 efficient biosynthesis 84:105 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 9 29 theme Toll-like 1415:1423 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 8 30 theme circular 1137:1144 arg1 spectroscopy 1156:1167 circular dichroism spectroscopy 1137:1167 circular dichroism spectroscopy 1137:1167 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 5 31 gly glycoprotein 834:845 arg1 glycoprotein 834:845 platelet glycoprotein Ib 825:848 platelet glycoprotein Ib 825:848 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 2 32 dep structures 381:390 arg1 response 339:346 response 339:346 response 339:346 In mammals Toll-like receptors signal in response to pathogen-associated microbial structures. 15173186 0 33 theme human 37:41 arg1 receptor 53:60 human toll-like receptor 2 37:62 human toll-like receptor 2 37:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 7 34 theme receptor 1009:1016 arg1 glycosylation 1018:1030 Toll-like receptor glycosylation 999:1030 Toll-like receptor glycosylation 999:1030 Toll-like receptor glycosylation is also likely to influence receptor surface representation, trafficking, and pattern recognition. 15173186 4 35 theme structural 580:589 arg1 principles 591:600 the structural principles 576:600 the structural principles that underlie recognition of these structures 576:646 However, the structural principles that underlie recognition of these structures are poorly understood. 15173186 0 36 theme glycosylation 14:26 arg1 sites 28:32 Four N-linked glycosylation sites 0:32 Four N-linked glycosylation sites in human toll-like receptor 2 0:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 1 37 theme Toll-like 277:285 arg1 receptors 287:295 so-called Toll-like receptors 267:295 so-called Toll-like receptors 267:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 1 37 theme Toll-like 277:285 arg1 receptors 256:264 evolutionarily related, germ line-encoded receptors 214:264 evolutionarily related, germ line-encoded receptors 214:264 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 6 38 theme Toll-like 941:949 arg1 receptor 951:958 Toll-like receptor 4 941:960 Toll-like receptor 4 941:960 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 11 39 theme site-directed 1581:1593 arg1 mutagenesis 1595:1605 site-directed mutagenesis 1581:1605 site-directed mutagenesis 1581:1605 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 3 40 theme bacterial 459:467 arg1 peptidoglycan 469:481 bacterial peptidoglycan 459:481 bacterial peptidoglycan 459:481 For example, Toll-like receptor 2 appears to mediate responses to bacterial peptidoglycan and acylated lipoproteins and Toll-like receptor 4 to bacterial lipopolysaccharide. 15173186 8 41 theme purified 1188:1195 arg1 proteins 1230:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 3 42 theme Toll-like 406:414 arg1 receptor 416:423 Toll-like receptor 2 406:425 Toll-like receptor 2 406:425 For example, Toll-like receptor 2 appears to mediate responses to bacterial peptidoglycan and acylated lipoproteins and Toll-like receptor 4 to bacterial lipopolysaccharide. 15173186 3 43 theme Toll-like 513:521 arg1 receptor 523:530 Toll-like receptor 4 513:532 Toll-like receptor 4 to bacterial lipopolysaccharide 513:564 For example, Toll-like receptor 2 appears to mediate responses to bacterial peptidoglycan and acylated lipoproteins and Toll-like receptor 4 to bacterial lipopolysaccharide. 15173186 0 44 theme toll-like 43:51 arg1 receptor 53:60 human toll-like receptor 2 37:62 human toll-like receptor 2 37:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 5 45 from repeats 709:715 arg1 domains 740:746 their extracellular domains 720:746 their extracellular domains 720:746 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 8 46 theme Toll-like 1203:1211 arg1 receptor 1213:1220 human Toll-like receptor 2 and 4 1197:1228 receptor 1213:1220 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 6 47 theme consensus 920:928 arg1 sites 930:934 N-linked glycosylation consensus sites 897:934 N-linked glycosylation consensus sites 897:934 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 7 48 theme Toll-like 999:1007 arg1 glycosylation 1018:1030 Toll-like receptor glycosylation 999:1030 Toll-like receptor glycosylation 999:1030 Toll-like receptor glycosylation is also likely to influence receptor surface representation, trafficking, and pattern recognition. 15173186 6 49 theme N-linked 897:904 arg1 sites 930:934 N-linked glycosylation consensus sites 897:934 N-linked glycosylation consensus sites 897:934 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 6 50 link N-linked 897:904 arg1 sites 930:934 N-linked glycosylation consensus sites 897:934 N-linked glycosylation consensus sites 897:934 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 8 51 theme 4 1228:1228 arg1 proteins 1230:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 10 52 theme significant 1460:1470 arg1 differences 1472:1482 significant differences 1460:1482 significant differences in the location and degree of conservation between sites in different Toll-like receptors 1460:1572 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 2 53 theme pathogen-associated 351:369 arg1 structures 381:390 pathogen-associated microbial structures 351:390 pathogen-associated microbial structures 351:390 In mammals Toll-like receptors signal in response to pathogen-associated microbial structures. 15173186 10 54 dep location 1491:1498 arg1 the 1487:1489 the 1487:1489 the 1487:1489 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 10 55 from sites 1535:1539 arg1 receptors 1564:1572 different Toll-like receptors 1544:1572 different Toll-like receptors 1544:1572 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 5 56 contain have 691:694 arg1 receptors 681:689 Toll-like receptors 671:689 Toll-like receptors 671:689 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 5 56 contain have 691:694 arg2 repeats 709:715 leucine-rich repeats 696:715 leucine-rich repeats in their extracellular domains 696:746 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 6 57 theme Toll-like 869:877 arg1 receptors 879:887 all Toll-like receptors 865:887 all Toll-like receptors 865:887 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 8 58 theme structure 1254:1262 arg1 contents 1264:1271 secondary structure contents 1244:1271 secondary structure contents similar to glycoprotein Ib 1244:1298 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 0 59 link N-linked 5:12 arg1 sites 28:32 Four N-linked glycosylation sites 0:32 Four N-linked glycosylation sites in human toll-like receptor 2 0:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 5 60 theme Toll-like 671:679 arg1 receptors 681:689 Toll-like receptors 671:689 Toll-like receptors 671:689 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 1 61 theme innate 161:166 arg1 defense 175:181 innate immune defense 161:181 innate immune defense 161:181 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 8 62 theme similar 1273:1279 arg1 contents 1264:1271 secondary structure contents 1244:1271 secondary structure contents similar to glycoprotein Ib 1244:1298 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 12 63 theme efficient 1902:1910 arg1 secretion 1920:1928 efficient protein secretion 1902:1928 efficient protein secretion 1902:1928 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 1 64 contain have 144:147 arg2 system 151:156 a system 149:156 a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors 149:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 1 64 contain have 144:147 arg1 organisms 134:142 Most higher organisms 122:142 Most higher organisms 122:142 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 2 65 theme Toll-like 309:317 arg1 receptors 319:327 Toll-like receptors 309:327 Toll-like receptors 309:327 In mammals Toll-like receptors signal in response to pathogen-associated microbial structures. 15173186 1 66 theme defense 175:181 arg1 system 151:156 a system 149:156 a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors 149:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 8 67 theme glycoprotein 1284:1295 arg1 Ib 1297:1298 glycoprotein Ib 1284:1298 glycoprotein Ib 1284:1298 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 10 68 theme conservation 1514:1525 arg1 degree 1504:1509 degree 1504:1509 degree 1504:1509 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 10 68 theme conservation 1514:1525 arg1 location 1491:1498 location 1491:1498 location 1491:1498 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 9 69 theme glycosylation 1339:1351 arg1 sites 1353:1357 consensus glycosylation sites 1329:1357 consensus glycosylation sites 1329:1357 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 11 70 theme glycosylation 1691:1703 arg1 sites 1705:1709 all four predicted glycosylation sites 1672:1709 all four predicted glycosylation sites 1672:1709 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 1 71 theme higher 127:132 arg1 organisms 134:142 Most higher organisms 122:142 Most higher organisms 122:142 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 12 72 theme glycosylation 1863:1875 arg1 sites 1877:1881 Toll-like receptor 2 glycosylation sites 1842:1881 The remaining Toll-like receptor 2 glycosylation sites 1828:1881 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 9 73 theme extracellular 1378:1390 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 9 73 theme extracellular 1378:1390 arg1 domains 1392:1398 the extracellular domains 1374:1398 the extracellular domains of other human Toll-like receptors 1374:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 0 74 from sites 28:32 arg1 receptor 53:60 human toll-like receptor 2 37:62 human toll-like receptor 2 37:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 11 75 theme receptor 1640:1647 arg1 domain 1665:1670 Toll-like receptor 2 extracellular domain 1630:1670 Toll-like receptor 2 extracellular domain 1630:1670 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 11 76 gly glycosylation 1691:1703 arg2 sites 1705:1709 all four predicted glycosylation sites 1672:1709 all four predicted glycosylation sites 1672:1709 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 11 76 gly glycosylation 1691:1703 arg2 four 1676:1679 four 1676:1679 four 1676:1679 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 12 77 theme remaining 1832:1840 arg1 sites 1877:1881 Toll-like receptor 2 glycosylation sites 1842:1881 The remaining Toll-like receptor 2 glycosylation sites 1828:1881 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 9 78 theme human 1409:1413 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 8 79 theme dichroism 1146:1154 arg1 spectroscopy 1156:1167 circular dichroism spectroscopy 1137:1167 circular dichroism spectroscopy 1137:1167 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 7 80 theme receptor 1060:1067 arg1 representation 1077:1090 receptor surface representation 1060:1090 receptor surface representation 1060:1090 Toll-like receptor glycosylation is also likely to influence receptor surface representation, trafficking, and pattern recognition. 15173186 12 81 theme receptor 1852:1859 arg1 sites 1877:1881 Toll-like receptor 2 glycosylation sites 1842:1881 The remaining Toll-like receptor 2 glycosylation sites 1828:1881 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 10 82 theme different 1544:1552 arg1 receptors 1564:1572 different Toll-like receptors 1544:1572 different Toll-like receptors 1544:1572 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 6 83 gly glycosylation 906:918 arg2 sites 930:934 N-linked glycosylation consensus sites 897:934 N-linked glycosylation consensus sites 897:934 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 5 84 theme similar 800:806 arg1 structures 788:797 solenoid structures 779:797 solenoid structures 779:797 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 9 85 theme receptors 1425:1433 arg1 receptors 1425:1433 other human Toll-like receptors 1403:1433 other human Toll-like receptors 1403:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 9 85 theme receptors 1425:1433 arg1 domains 1392:1398 the extracellular domains 1374:1398 the extracellular domains of other human Toll-like receptors 1374:1433 We have also analyzed where consensus glycosylation sites are located in the extracellular domains of other human Toll-like receptors. 15173186 0 86 theme N-linked 5:12 arg1 sites 28:32 Four N-linked glycosylation sites 0:32 Four N-linked glycosylation sites in human toll-like receptor 2 0:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 1 87 theme so-called 267:275 arg1 receptors 287:295 so-called Toll-like receptors 267:295 so-called Toll-like receptors 267:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 1 87 theme so-called 267:275 arg1 receptors 256:264 evolutionarily related, germ line-encoded receptors 214:264 evolutionarily related, germ line-encoded receptors 214:264 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 10 88 from differences 1472:1482 arg1 degree 1504:1509 degree 1504:1509 degree 1504:1509 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 10 88 from differences 1472:1482 arg1 location 1491:1498 location 1491:1498 location 1491:1498 We found that there are significant differences in the location and degree of conservation between sites in different Toll-like receptors. 15173186 3 89 theme acylated 487:494 arg1 lipoproteins 496:507 acylated lipoproteins 487:507 acylated lipoproteins 487:507 For example, Toll-like receptor 2 appears to mediate responses to bacterial peptidoglycan and acylated lipoproteins and Toll-like receptor 4 to bacterial lipopolysaccharide. 15173186 5 90 theme glycoprotein 834:845 arg1 Ib 847:848 platelet glycoprotein Ib 825:848 platelet glycoprotein Ib 825:848 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 8 91 theme human 1197:1201 arg1 receptor 1213:1220 human Toll-like receptor 2 and 4 1197:1228 receptor 1213:1220 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 8 92 theme receptor 1213:1220 arg1 proteins 1230:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 0 93 gly glycosylation 14:26 arg2 Four 0:3 Four 0:3 Four 0:3 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 0 93 gly glycosylation 14:26 arg2 sites 28:32 Four N-linked glycosylation sites 0:32 Four N-linked glycosylation sites in human toll-like receptor 2 0:62 Four N-linked glycosylation sites in human toll-like receptor 2 cooperate to direct efficient biosynthesis and secretion. 15173186 8 94 contain have 1239:1242 arg2 contents 1264:1271 secondary structure contents 1244:1271 secondary structure contents similar to glycoprotein Ib 1244:1298 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 8 94 contain have 1239:1242 arg1 proteins 1230:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 purified human Toll-like receptor 2 and 4 proteins 1188:1237 Using circular dichroism spectroscopy, we show here that purified human Toll-like receptor 2 and 4 proteins have secondary structure contents similar to glycoprotein Ib. 15173186 6 95 theme glycosylation 906:918 arg1 sites 930:934 N-linked glycosylation consensus sites 897:934 N-linked glycosylation consensus sites 897:934 Additionally, all Toll-like receptors contain N-linked glycosylation consensus sites, and Toll-like receptor 4 requires glycosylation for function. 15173186 5 96 located found 816:820 arg1 Ib 847:848 platelet glycoprotein Ib 825:848 platelet glycoprotein Ib 825:848 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 5 96 located found 816:820 arg2 that 811:814 that 811:814 that 811:814 Toll-like receptors have leucine-rich repeats in their extracellular domains and are thus believed to adopt solenoid structures, similar to that found in platelet glycoprotein Ib. 15173186 2 97 theme microbial 371:379 arg1 structures 381:390 pathogen-associated microbial structures 351:390 pathogen-associated microbial structures 351:390 In mammals Toll-like receptors signal in response to pathogen-associated microbial structures. 15173186 12 98 theme lesser 1943:1948 arg1 degree 1950:1955 a lesser degree 1941:1955 a lesser degree 1941:1955 The remaining Toll-like receptor 2 glycosylation sites also contribute to efficient protein secretion, albeit to a lesser degree. 15173186 11 99 gly core-glycosylated 1763:1779 arg1 site 1741:1744 one site 1737:1744 one site 1737:1744 Using site-directed mutagenesis, we have found that in Toll-like receptor 2 extracellular domain all four predicted glycosylation sites are substituted, although one site is inefficiently core-glycosylated and its removal drastically affects secretion. 15173186 1 100 theme related 229:235 arg1 receptors 287:295 so-called Toll-like receptors 267:295 so-called Toll-like receptors 267:295 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 15173186 1 100 theme related 229:235 arg1 receptors 256:264 evolutionarily related, germ line-encoded receptors 214:264 evolutionarily related, germ line-encoded receptors 214:264 Most higher organisms have a system of innate immune defense that is mediated by a group of evolutionarily related, germ line-encoded receptors, so-called Toll-like receptors. 20826823 0 0 theme N-glycosylation 67:81 arg1 role 59:62 the role 55:62 the role of N-glycosylation 55:81 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 0 theme N-glycosylation 67:81 arg1 structure 99:107 the crystal structure 87:107 the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407 87:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 1 1 theme kallikrein-kinin 326:341 arg1 systems 343:349 the renin-angiotensin and kallikrein-kinin systems 300:349 the renin-angiotensin and kallikrein-kinin systems 300:349 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 8 2 from structure 1302:1310 arg1 presence 1319:1326 the presence 1315:1326 the presence of an N domain-selective phosphinic inhibitor RXP407 1315:1379 The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. 20826823 4 3 theme mass 809:812 arg1 spectrometry 814:825 mass spectrometry 809:825 mass spectrometry 809:825 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 5 4 theme enzymatic 949:957 arg1 activity 959:966 enzymatic activity 949:966 enzymatic activity 949:966 A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. 20826823 4 5 theme sites 745:749 arg1 importance 698:707 the importance 694:707 the importance of the 10 potential N-linked glycan sites 694:749 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 6 6 gly glycosylated 1046:1057 arg1 sites 1036:1040 the potential glycan sites 1015:1040 the potential glycan sites 1015:1040 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 6 gly glycosylated 1046:1057 arg1 10 1009:1010 10 1009:1010 10 1009:1010 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 6 gly glycosylated 1046:1057 arg1 eight 1000:1004 eight 1000:1004 eight 1000:1004 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 7 theme domain 1126:1131 arg1 expression 1103:1112 expression 1103:1112 expression of active N domain 1103:1131 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 4 8 theme potential 719:727 arg1 sites 745:749 the 10 potential N-linked glycan sites 712:749 the 10 potential N-linked glycan sites 712:749 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 8 9 theme domain-selective 1336:1351 arg1 RXP407 1374:1379 an N domain-selective phosphinic inhibitor RXP407 1331:1379 an N domain-selective phosphinic inhibitor RXP407 1331:1379 The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. 20826823 6 10 theme active 1117:1122 arg1 domain 1126:1131 active N domain 1117:1131 active N domain 1117:1131 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 1 11 from role 292:295 arg1 systems 343:349 the renin-angiotensin and kallikrein-kinin systems 300:349 the renin-angiotensin and kallikrein-kinin systems 300:349 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 3 12 theme surface 519:525 arg1 glycosylation 527:539 surface glycosylation 519:539 surface glycosylation of the protein 519:554 Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. 20826823 5 13 theme site-directed 881:893 arg1 mutagenesis 895:905 site-directed mutagenesis 881:905 site-directed mutagenesis 881:905 A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. 20826823 4 14 gly glycosylation 593:605 arg1 N 614:614 the N domain 610:621 the N domain 610:621 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 4 15 theme glycosylation 593:605 arg1 role 585:588 the role 581:588 the role of glycosylation in the N domain 581:621 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 4 16 theme glycan 738:743 arg1 sites 745:749 the 10 potential N-linked glycan sites 712:749 the 10 potential N-linked glycan sites 712:749 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 2 17 theme N 382:382 arg1 domains 369:375 two domains 365:375 two domains 365:375 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 2 17 theme N 382:382 arg1 domains 390:396 the N and C domains 378:396 the N and C domains 378:396 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 0 18 theme crystal 91:97 arg1 structure 99:107 the crystal structure 87:107 the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407 87:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 2 19 contain contains 356:363 arg1 ACE 352:354 ACE 352:354 ACE 352:354 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 2 19 contain contains 356:363 arg2 domains 390:396 the N and C domains 378:396 the N and C domains 378:396 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 2 19 contain contains 356:363 arg2 domains 369:375 two domains 365:375 two domains 365:375 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 4 20 theme N-linked 729:736 arg1 sites 745:749 the 10 potential N-linked glycan sites 712:749 the 10 potential N-linked glycan sites 712:749 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 6 21 theme sites 1036:1040 arg1 sites 1036:1040 the potential glycan sites 1015:1040 the potential glycan sites 1015:1040 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 21 theme sites 1036:1040 arg1 10 1009:1010 10 1009:1010 10 1009:1010 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 0 22 theme N 4:4 arg1 domain 6:11 The N domain 0:11 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 0:174 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 22 theme N 4:4 arg1 enzyme 47:52 human angiotensin-I-converting enzyme 16:52 human angiotensin-I-converting enzyme 16:52 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 7 23 theme glycosylated 1218:1229 arg1 construct 1239:1247 The minimally glycosylated Ndom389 construct 1204:1247 The minimally glycosylated Ndom389 construct 1204:1247 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 7 23 theme glycosylated 1218:1229 arg1 suitable 1260:1267 suitable 1260:1267 suitable 1260:1267 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 6 24 theme N 1124:1124 arg1 domain 1126:1131 active N domain 1117:1131 active N domain 1117:1131 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 25 theme glycan 1029:1034 arg1 sites 1036:1040 the potential glycan sites 1015:1040 the potential glycan sites 1015:1040 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 1 26 theme blood 257:261 arg1 pressure 263:270 blood pressure 257:270 blood pressure 257:270 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 3 27 theme protein 548:554 arg1 glycosylation 527:539 surface glycosylation 519:539 surface glycosylation of the protein 519:554 Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. 20826823 1 28 theme Angiotensin-I-converting 176:199 arg1 ACE 209:211 ACE 209:211 ACE 209:211 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 1 28 theme Angiotensin-I-converting 176:199 arg1 enzyme 201:206 Angiotensin-I-converting enzyme 176:206 Angiotensin-I-converting enzyme (ACE) 176:212 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 1 29 theme pressure 263:270 arg1 regulation 243:252 the regulation 239:252 the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems 239:349 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 7 30 gly glycosylated 1218:1229 arg1 construct 1239:1247 The minimally glycosylated Ndom389 construct 1204:1247 The minimally glycosylated Ndom389 construct 1204:1247 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 7 30 gly glycosylated 1218:1229 arg1 suitable 1260:1267 suitable 1260:1267 suitable 1260:1267 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 6 31 theme 10 1009:1010 arg1 sites 1036:1040 the potential glycan sites 1015:1040 the potential glycan sites 1015:1040 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 31 theme 10 1009:1010 arg1 10 1009:1010 10 1009:1010 10 1009:1010 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 6 31 theme 10 1009:1010 arg1 eight 1000:1004 eight 1000:1004 eight 1000:1004 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 7 32 theme crystallization 1273:1287 arg1 studies 1289:1295 crystallization studies 1273:1295 crystallization studies 1273:1295 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 5 33 theme CHO 921:923 arg1 cells 925:929 CHO cells 921:929 CHO cells 921:929 A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. 20826823 8 34 theme RXP407 1374:1379 arg1 presence 1319:1326 the presence 1315:1326 the presence of an N domain-selective phosphinic inhibitor RXP407 1315:1379 The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. 20826823 8 35 theme phosphinic 1353:1362 arg1 RXP407 1374:1379 an N domain-selective phosphinic inhibitor RXP407 1331:1379 an N domain-selective phosphinic inhibitor RXP407 1331:1379 The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. 20826823 3 36 theme Structural 439:448 arg1 studies 450:456 Structural studies 439:456 Structural studies of ACE 439:463 Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. 20826823 0 37 theme angiotensin-I-converting 22:45 arg1 enzyme 47:52 human angiotensin-I-converting enzyme 16:52 human angiotensin-I-converting enzyme 16:52 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 4 38 theme N 614:614 arg1 domain 616:621 the N domain 610:621 the N domain 610:621 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 4 39 theme limited 784:790 arg1 proteolysis 792:802 limited proteolysis 784:802 limited proteolysis 784:802 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 5 40 theme glycosylation 840:852 arg1 mutants 854:860 glycosylation mutants 840:860 glycosylation mutants 840:860 A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. 20826823 2 41 theme C 388:388 arg1 domains 369:375 two domains 365:375 two domains 365:375 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 2 41 theme C 388:388 arg1 domains 390:396 the N and C domains 378:396 the N and C domains 378:396 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 0 42 theme human 16:20 arg1 enzyme 47:52 human angiotensin-I-converting enzyme 16:52 human angiotensin-I-converting enzyme 16:52 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 1 43 theme central 284:290 arg1 role 292:295 its central role 280:295 its central role in the renin-angiotensin and kallikrein-kinin systems 280:349 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 0 44 from role 59:62 arg1 complex 112:118 complex 112:118 complex with an N domain-specific phosphinic inhibitor, RXP407 112:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 6 45 theme thermal 1185:1191 arg1 stability 1193:1201 its thermal stability 1181:1201 its thermal stability 1181:1201 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 0 46 theme domain-specific 130:144 arg1 RXP407 168:173 RXP407 168:173 RXP407 168:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 46 theme domain-specific 130:144 arg1 inhibitor 157:165 an N domain-specific phosphinic inhibitor 125:165 an N domain-specific phosphinic inhibitor 125:165 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 6 47 theme C-terminal 1066:1075 arg1 sites 1077:1081 three C-terminal sites 1060:1081 three C-terminal sites 1060:1081 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 3 48 theme ACE 461:463 arg1 studies 450:456 Structural studies 439:456 Structural studies of ACE 439:463 Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. 20826823 0 49 theme enzyme 47:52 arg1 domain 6:11 The N domain 0:11 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 0:174 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 49 theme enzyme 47:52 arg1 enzyme 47:52 human angiotensin-I-converting enzyme 16:52 human angiotensin-I-converting enzyme 16:52 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 6 50 theme potential 1019:1027 arg1 sites 1036:1040 the potential glycan sites 1015:1040 the potential glycan sites 1015:1040 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 0 51 from structure 99:107 arg1 complex 112:118 complex 112:118 complex with an N domain-specific phosphinic inhibitor, RXP407 112:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 2 52 gly glycosylated 425:436 arg1 both 399:402 both 399:402 both 399:402 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 2 52 gly glycosylated 425:436 arg1 domains 390:396 the N and C domains 378:396 the N and C domains 378:396 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 2 52 gly glycosylated 425:436 arg1 domains 369:375 two domains 365:375 two domains 365:375 ACE contains two domains, the N and C domains, both of which are heavily glycosylated. 20826823 9 53 theme hinge 1450:1454 arg1 region 1456:1461 a hinge region 1448:1461 a hinge region that may contribute to the breathing motion proposed for substrate binding 1448:1536 The Ndom389 structure revealed a hinge region that may contribute to the breathing motion proposed for substrate binding. 20826823 4 54 link N-linked 729:736 arg1 sites 745:749 the 10 potential N-linked glycan sites 712:749 the 10 potential N-linked glycan sites 712:749 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 6 55 theme N-terminal 1146:1155 arg1 sites 1157:1161 two N-terminal sites 1142:1161 two N-terminal sites 1142:1161 At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. 20826823 8 56 theme Å 1403:1403 arg1 resolution 1405:1414 2.0 Å resolution 1399:1414 2.0 Å resolution 1399:1414 The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. 20826823 4 57 theme suitable 637:644 arg1 forms 646:650 suitable forms 637:650 suitable forms for crystallization 637:670 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 7 58 theme Ndom389 1231:1237 arg1 construct 1239:1247 The minimally glycosylated Ndom389 construct 1204:1247 The minimally glycosylated Ndom389 construct 1204:1247 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 7 58 theme Ndom389 1231:1237 arg1 suitable 1260:1267 suitable 1260:1267 suitable 1260:1267 The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. 20826823 8 59 theme inhibitor 1364:1372 arg1 RXP407 1374:1379 an N domain-selective phosphinic inhibitor RXP407 1331:1379 an N domain-selective phosphinic inhibitor RXP407 1331:1379 The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. 20826823 0 60 theme phosphinic 146:155 arg1 RXP407 168:173 RXP407 168:173 RXP407 168:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 60 theme phosphinic 146:155 arg1 inhibitor 157:165 an N domain-specific phosphinic inhibitor 125:165 an N domain-specific phosphinic inhibitor 125:165 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 9 61 theme substrate 1520:1528 arg1 binding 1530:1536 substrate binding 1520:1536 substrate binding 1520:1536 The Ndom389 structure revealed a hinge region that may contribute to the breathing motion proposed for substrate binding. 20826823 0 62 dep domain 6:11 arg1 role 59:62 the role 55:62 the role of N-glycosylation 55:81 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 62 dep domain 6:11 arg1 structure 99:107 the crystal structure 87:107 the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407 87:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 3 63 gly glycosylation 527:539 arg1 protein 548:554 the protein 544:554 the protein 544:554 Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. 20826823 0 64 with complex 112:118 arg1 RXP407 168:173 RXP407 168:173 RXP407 168:173 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 0 64 with complex 112:118 arg1 inhibitor 157:165 an N domain-specific phosphinic inhibitor 125:165 an N domain-specific phosphinic inhibitor 125:165 The N domain of human angiotensin-I-converting enzyme: the role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407. 20826823 5 65 theme thermal 972:978 arg1 stability 980:988 thermal stability 972:988 thermal stability 972:988 A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. 20826823 9 66 theme breathing 1490:1498 arg1 motion 1500:1505 the breathing motion 1486:1505 the breathing motion proposed for substrate binding 1486:1536 The Ndom389 structure revealed a hinge region that may contribute to the breathing motion proposed for substrate binding. 20826823 5 67 theme mutants 854:860 arg1 number 830:835 A number 828:835 A number of glycosylation mutants 828:860 A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. 20826823 9 68 theme Ndom389 1421:1427 arg1 structure 1429:1437 The Ndom389 structure 1417:1437 The Ndom389 structure 1417:1437 The Ndom389 structure revealed a hinge region that may contribute to the breathing motion proposed for substrate binding. 20826823 1 69 theme critical 222:229 arg1 role 231:234 a critical role 220:234 a critical role 220:234 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 4 70 from role 585:588 arg1 domain 616:621 the N domain 610:621 the N domain 610:621 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 1 71 theme renin-angiotensin 304:320 arg1 systems 343:349 the renin-angiotensin and kallikrein-kinin systems 300:349 the renin-angiotensin and kallikrein-kinin systems 300:349 Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. 20826823 4 72 theme enzymatic 757:765 arg1 deglycosylation 767:781 enzymatic deglycosylation 757:781 enzymatic deglycosylation 757:781 In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. 20826823 3 73 theme severe 488:493 arg1 difficulties 495:506 severe difficulties 488:506 severe difficulties 488:506 Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. 22448645 0 0 theme 5-HT₇a 87:92 arg1 receptor 94:101 the human serotonin 5-HT₇a receptor 67:101 the human serotonin 5-HT₇a receptor 67:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 2 1 theme N-glycosylation 325:339 arg1 role 317:320 the functional role 302:320 the functional role of N-glycosylation 302:339 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 2 1 theme N-glycosylation 325:339 arg1 presence 289:296 presence 289:296 presence 289:296 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 1 2 theme neurological 227:238 arg1 disorders 240:248 human psychiatric and neurological disorders 205:248 human psychiatric and neurological disorders 205:248 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 0 3 theme serotonin 77:85 arg1 receptor 94:101 the human serotonin 5-HT₇a receptor 67:101 the human serotonin 5-HT₇a receptor 67:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 8 4 theme 5-HT 1353:1356 arg1 receptor 1361:1368 the 5-HT(7) receptor 1349:1368 the 5-HT(7) receptor 1349:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 4 5 dep mutated 627:633 arg1 N66Q 684:687 N66Q 684:687 N66Q 684:687 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 4 5 dep mutated 627:633 arg1 N5Q 676:678 N5Q 676:678 N5Q 676:678 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 5 6 contain had 863:865 arg1 receptors 853:861 The mutant receptors 842:861 The mutant receptors 842:861 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 5 6 contain had 863:865 arg2 affinity 884:891 the same binding affinity 867:891 the same binding affinity for [(3) H]5-CT 867:907 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 5 6 contain had 863:865 arg2 potency 922:928 same potency 917:928 same potency 917:928 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 5 7 dep H 902:902 arg1 3 899:899 3 899:899 3 899:899 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 4 8 theme wild-type 768:776 arg1 receptor 778:785 the wild-type receptor 764:785 the wild-type receptor 764:785 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 5 9 theme 5-HT-induced 958:969 arg1 activation 971:980 5-HT-induced activation 958:980 5-HT-induced activation of adenylyl cyclase 958:1000 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 4 10 theme bands 744:748 arg1 mass 717:720 the molecular mass 703:720 the molecular mass of the immunoreactive bands 703:748 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 4 11 gly N-glycosylated 826:839 arg1 asparagines 809:819 both asparagines 804:819 both asparagines 804:819 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 6 12 from reduction 1024:1032 arg1 binding 1052:1058 maximal ligand binding 1037:1058 maximal ligand binding 1037:1058 However, there was a reduction in maximal ligand binding for the single and double mutants compared to the wild-type receptor. 22448645 3 13 gly N-glycosylated 547:560 arg1 form 562:565 an N-glycosylated form 544:565 an N-glycosylated form of the 5-HT(7(a)) receptor 544:592 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 3 13 gly N-glycosylated 547:560 arg1 receptor 585:592 the 5-HT(7(a)) receptor 570:592 the 5-HT(7(a)) receptor 570:592 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 0 14 theme receptor 94:101 arg1 glycosylation 50:62 N-linked glycosylation 41:62 N-linked glycosylation of the human serotonin 5-HT₇a receptor 41:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 5 15 theme same 917:920 arg1 potency 922:928 same potency 917:928 same potency 917:928 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 4 16 theme immunoreactive 729:742 arg1 bands 744:748 the immunoreactive bands 725:748 the immunoreactive bands 725:748 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 6 17 theme double 1079:1084 arg1 mutants 1086:1092 the single and double mutants 1064:1092 the single and double mutants 1064:1092 However, there was a reduction in maximal ligand binding for the single and double mutants compared to the wild-type receptor. 22448645 2 18 theme present 258:264 arg1 study 266:270 the present study 254:270 the present study 254:270 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 3 19 theme blot 380:383 arg1 analysis 385:392 Western blot analysis 372:392 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin 372:487 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 8 20 theme cell 1322:1325 arg1 expression 1335:1344 cell surface expression 1322:1344 cell surface expression of the 5-HT(7) receptor 1322:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 3 21 theme receptor 585:592 arg1 form 562:565 an N-glycosylated form 544:565 an N-glycosylated form of the 5-HT(7(a)) receptor 544:592 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 4 22 gly N-glycosylation 653:667 arg2 two 639:641 two 639:641 two 639:641 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 4 22 gly N-glycosylation 653:667 arg2 sites 669:673 the two predicted N-glycosylation sites 635:673 the two predicted N-glycosylation sites 635:673 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 5 23 theme mutant 846:851 arg1 receptors 853:861 The mutant receptors 842:861 The mutant receptors 842:861 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 3 24 theme HEK293T 397:403 arg1 cells 405:409 HEK293T cells 397:409 HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin 397:487 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 6 25 theme single 1068:1073 arg1 mutants 1086:1092 the single and double mutants 1064:1092 the single and double mutants 1064:1092 However, there was a reduction in maximal ligand binding for the single and double mutants compared to the wild-type receptor. 22448645 2 26 theme 5-HT 354:357 arg1 receptor 362:369 the human 5-HT(7) receptor 344:369 the human 5-HT(7) receptor 344:369 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 0 27 theme Biochemical 0:10 arg1 study 32:36 Biochemical and pharmacological study 0:36 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor 0:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 4 28 theme molecular 707:715 arg1 mass 717:720 the molecular mass 703:720 the molecular mass of the immunoreactive bands 703:748 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 0 29 link N-linked 41:48 arg1 glycosylation 50:62 N-linked glycosylation 41:62 N-linked glycosylation of the human serotonin 5-HT₇a receptor 41:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 7 30 theme cell 1254:1257 arg1 surface 1259:1265 the cell surface 1250:1265 the cell surface 1250:1265 Next, membrane labelling and immunocytochemical studies demonstrated that the N-glycosylation mutants were expressed at the cell surface. 22448645 2 31 theme human 348:352 arg1 receptor 362:369 the human 5-HT(7) receptor 344:369 the human 5-HT(7) receptor 344:369 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 7 32 theme immunocytochemical 1159:1176 arg1 studies 1178:1184 immunocytochemical studies 1159:1184 immunocytochemical studies 1159:1184 Next, membrane labelling and immunocytochemical studies demonstrated that the N-glycosylation mutants were expressed at the cell surface. 22448645 6 33 theme maximal 1037:1043 arg1 binding 1052:1058 maximal ligand binding 1037:1058 maximal ligand binding 1037:1058 However, there was a reduction in maximal ligand binding for the single and double mutants compared to the wild-type receptor. 22448645 3 34 theme tunicamycin 477:487 arg1 presence 465:472 the presence 461:472 the presence of tunicamycin 461:487 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 1 35 theme G-protein-coupled 155:171 arg1 receptor 173:180 a G-protein-coupled receptor 153:180 a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders 153:248 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 1 35 theme G-protein-coupled 155:171 arg1 receptor 141:148 The 5-hydroxytryptamine (5-HT)(7(a)) receptor 104:148 The 5-hydroxytryptamine (5-HT)(7(a)) receptor 104:148 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 0 36 theme pharmacological 16:30 arg1 study 32:36 Biochemical and pharmacological study 0:36 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor 0:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 4 37 theme predicted 643:651 arg1 sites 669:673 the two predicted N-glycosylation sites 635:673 the two predicted N-glycosylation sites 635:673 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 3 38 theme form 562:565 arg1 existence 531:539 the existence 527:539 the existence of an N-glycosylated form of the 5-HT(7(a)) receptor 527:592 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 1 39 theme 5-hydroxytryptamine 108:126 arg1 receptor 173:180 a G-protein-coupled receptor 153:180 a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders 153:248 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 1 39 theme 5-hydroxytryptamine 108:126 arg1 receptor 141:148 The 5-hydroxytryptamine (5-HT)(7(a)) receptor 104:148 The 5-hydroxytryptamine (5-HT)(7(a)) receptor 104:148 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 0 40 theme N-linked 41:48 arg1 glycosylation 50:62 N-linked glycosylation 41:62 N-linked glycosylation of the human serotonin 5-HT₇a receptor 41:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 3 41 theme cells 405:409 arg1 analysis 385:392 Western blot analysis 372:392 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin 372:487 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 5 42 theme binding 876:882 arg1 affinity 884:891 the same binding affinity 867:891 the same binding affinity for [(3) H]5-CT 867:907 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 7 43 theme membrane 1136:1143 arg1 labelling 1145:1153 membrane labelling 1136:1153 membrane labelling 1136:1153 Next, membrane labelling and immunocytochemical studies demonstrated that the N-glycosylation mutants were expressed at the cell surface. 22448645 2 44 theme receptor 362:369 arg1 role 317:320 the functional role 302:320 the functional role of N-glycosylation 302:339 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 2 44 theme receptor 362:369 arg1 presence 289:296 presence 289:296 presence 289:296 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 3 45 theme Western 372:378 arg1 analysis 385:392 Western blot analysis 372:392 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin 372:487 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 7 46 theme N-glycosylation 1208:1222 arg1 mutants 1224:1230 the N-glycosylation mutants 1204:1230 the N-glycosylation mutants 1204:1230 Next, membrane labelling and immunocytochemical studies demonstrated that the N-glycosylation mutants were expressed at the cell surface. 22448645 5 47 dep potency 922:928 arg1 the 913:915 the 913:915 the 913:915 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 3 48 theme N-glycosylated 547:560 arg1 form 562:565 an N-glycosylated form 544:565 an N-glycosylated form of the 5-HT(7(a)) receptor 544:592 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 6 49 theme ligand 1045:1050 arg1 binding 1052:1058 maximal ligand binding 1037:1058 maximal ligand binding 1037:1058 However, there was a reduction in maximal ligand binding for the single and double mutants compared to the wild-type receptor. 22448645 8 50 theme receptor 1361:1368 arg1 expression 1335:1344 cell surface expression 1322:1344 cell surface expression of the 5-HT(7) receptor 1322:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 0 51 theme glycosylation 50:62 arg1 study 32:36 Biochemical and pharmacological study 0:36 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor 0:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 5 52 theme adenylyl 985:992 arg1 cyclase 994:1000 adenylyl cyclase 985:1000 adenylyl cyclase 985:1000 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 3 53 theme 5-HT 438:441 arg1 receptor 449:456 the 5-HT(7(a)) receptor 434:456 the 5-HT(7(a)) receptor 434:456 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 3 54 theme band 504:507 arg1 shift 509:513 a band shift 502:513 a band shift 502:513 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 2 55 theme functional 306:315 arg1 role 317:320 the functional role 302:320 the functional role of N-glycosylation 302:339 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 5 56 theme cyclase 994:1000 arg1 activation 971:980 5-HT-induced activation 958:980 5-HT-induced activation of adenylyl cyclase 958:1000 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 22448645 2 57 dep presence 289:296 arg1 the 285:287 the 285:287 the 285:287 In the present study, we evaluate the presence and the functional role of N-glycosylation of the human 5-HT(7) receptor. 22448645 6 58 theme wild-type 1110:1118 arg1 receptor 1120:1127 the wild-type receptor 1106:1127 the wild-type receptor 1106:1127 However, there was a reduction in maximal ligand binding for the single and double mutants compared to the wild-type receptor. 22448645 8 59 theme surface 1327:1333 arg1 expression 1335:1344 cell surface expression 1322:1344 cell surface expression of the 5-HT(7) receptor 1322:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 4 60 theme N-glycosylation 653:667 arg1 sites 669:673 the two predicted N-glycosylation sites 635:673 the two predicted N-glycosylation sites 635:673 To further investigate this, we mutated the two predicted N-glycosylation sites (N5Q and N66Q) and compared the molecular mass of the immunoreactive bands with those of the wild-type receptor, indicating that both asparagines were N-glycosylated. 22448645 1 61 theme human 205:209 arg1 disorders 240:248 human psychiatric and neurological disorders 205:248 human psychiatric and neurological disorders 205:248 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 0 62 theme human 71:75 arg1 receptor 94:101 the human serotonin 5-HT₇a receptor 67:101 the human serotonin 5-HT₇a receptor 67:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 3 63 theme 5-HT 574:577 arg1 receptor 585:592 the 5-HT(7(a)) receptor 570:592 the 5-HT(7(a)) receptor 570:592 Western blot analysis of HEK293T cells transiently expressing the 5-HT(7(a)) receptor in the presence of tunicamycin gave rise to a band shift, indicating the existence of an N-glycosylated form of the 5-HT(7(a)) receptor. 22448645 0 64 gly glycosylation 50:62 arg1 receptor 94:101 the human serotonin 5-HT₇a receptor 67:101 the human serotonin 5-HT₇a receptor 67:101 Biochemical and pharmacological study of N-linked glycosylation of the human serotonin 5-HT₇a receptor. 22448645 8 65 gly N-glycosylation 1285:1299 arg1 surface 1327:1333 cell surface expression 1322:1344 cell surface expression of the 5-HT(7) receptor 1322:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 8 65 gly N-glycosylation 1285:1299 arg1 cell 1322:1325 cell surface expression 1322:1344 cell surface expression of the 5-HT(7) receptor 1322:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 8 65 gly N-glycosylation 1285:1299 arg1 receptor 1361:1368 the 5-HT(7) receptor 1349:1368 the 5-HT(7) receptor 1349:1368 We conclude that N-glycosylation is not important for cell surface expression of the 5-HT(7) receptor. 22448645 1 66 theme psychiatric 211:221 arg1 disorders 240:248 human psychiatric and neurological disorders 205:248 human psychiatric and neurological disorders 205:248 The 5-hydroxytryptamine (5-HT)(7(a)) receptor is a G-protein-coupled receptor critically involved in human psychiatric and neurological disorders. 22448645 5 67 theme same 871:874 arg1 affinity 884:891 the same binding affinity 867:891 the same binding affinity for [(3) H]5-CT 867:907 The mutant receptors had the same binding affinity for [(3) H]5-CT and the same potency and efficacy with regard to 5-HT-induced activation of adenylyl cyclase. 20837471 6 0 from Thr 1105:1107 arg1 ANGPTL3 1171:1177 ANGPTL3 1171:1177 ANGPTL3 1171:1177 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 4 1 theme Genome-wide 675:685 arg1 studies 703:709 Genome-wide SNP association studies 675:709 Genome-wide SNP association studies 675:709 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 1 2 theme endothelial 266:276 arg1 lipases 294:300 the endothelial and lipoprotein lipases 262:300 lipases 294:300 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 0 3 theme possible 91:98 arg1 role 100:103 possible role 91:103 possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids 91:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 2 4 theme RAPR 383:386 arg1 processing 371:380 proprotein convertase processing 349:380 proprotein convertase processing (RAPR(224)↓TT) for activation 349:410 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 4 theme RAPR 383:386 arg1 ↓TT 392:394 RAPR(224)↓TT 383:394 RAPR(224)↓TT 383:394 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 6 5 theme panel 983:987 arg1 Screening 968:976 Screening 968:976 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 968:1078 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 6 6 theme proteins 992:999 arg1 panel 983:987 a panel 981:987 a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 981:1078 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 0 7 theme polypeptide 108:118 arg1 GalNAc-transferase-2 120:139 polypeptide GalNAc-transferase-2 108:139 polypeptide GalNAc-transferase-2 108:139 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 3 8 theme ANGPTL3 666:672 arg1 processing 652:661 processing 652:661 processing of ANGPTL3 652:672 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 2 9 contain contains 437:444 arg1 site 432:435 the processing site 417:435 the processing site 417:435 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 9 contain contains 437:444 arg2 sites 483:487 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 4 10 theme polypeptide 731:741 arg1 GalNAc-transferase 743:760 polypeptide GalNAc-transferase 731:760 the polypeptide GalNAc-transferase gene 727:765 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 1 11 theme lipoprotein 282:292 arg1 lipases 294:300 the endothelial and lipoprotein lipases 262:300 lipases 294:300 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 3 12 from O-glycosylation 605:619 arg1 site 639:642 the processing site 624:642 the processing site 624:642 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 2 13 dep sites 483:487 arg1 TT 513:514 TT(226) 513:519 TT(226) 513:519 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 13 dep sites 483:487 arg1 C-terminal 501:510 C-terminal 501:510 C-terminal 501:510 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 3 14 theme in 539:540 arg1 system 553:558 an in vivo model system 536:558 an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3 536:672 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 1 15 theme lipases 294:300 arg1 inhibitor 249:257 an important inhibitor 236:257 an important inhibitor of the endothelial and lipoprotein lipases 236:300 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 1 15 theme lipases 294:300 arg1 target 323:328 a promising drug target 306:328 a promising drug target 306:328 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 1 15 theme lipases 294:300 arg1 protein 213:219 angiopoietin-like protein 3 195:221 The angiopoietin-like protein 3 (ANGPTL3) 191:231 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 5 16 theme critical 901:908 arg1 O-glycosylation 910:924 critical O-glycosylation 901:924 critical O-glycosylation of proteins involved in lipid metabolism 901:965 We hypothesized that the GalNAc-T2 transferase performed critical O-glycosylation of proteins involved in lipid metabolism. 20837471 4 17 theme triglyceride 817:828 arg1 levels 836:841 high triglyceride blood levels 812:841 high triglyceride blood levels 812:841 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 1 18 theme angiopoietin-like 195:211 arg1 protein 213:219 angiopoietin-like protein 3 195:221 The angiopoietin-like protein 3 (ANGPTL3) 191:231 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 1 18 theme angiopoietin-like 195:211 arg1 ANGPTL3 224:230 ANGPTL3 224:230 ANGPTL3 224:230 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 1 18 theme angiopoietin-like 195:211 arg1 inhibitor 249:257 an important inhibitor 236:257 an important inhibitor of the endothelial and lipoprotein lipases 236:300 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 0 19 theme GalNAc-transferase-2 120:139 arg1 role 100:103 possible role 91:103 possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids 91:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 4 20 theme high 812:815 arg1 levels 836:841 high triglyceride blood levels 812:841 high triglyceride blood levels 812:841 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 6 21 theme proprotein 1130:1139 arg1 processing 1152:1161 the proprotein convertase processing 1126:1161 the proprotein convertase processing 1126:1161 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 6 22 theme lipid 1017:1021 arg1 metabolism 1023:1032 lipid metabolism 1017:1032 lipid metabolism for potential sites glycosylated by GalNAc-T2 1017:1078 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 7 23 theme in 1295:1296 arg1 cleavage 1310:1317 in vitro furin cleavage 1295:1317 in vitro furin cleavage 1295:1317 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 2 24 theme potential 450:458 arg1 sites 483:487 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 3 25 gly O-glycosylation 605:619 arg1 site 639:642 the processing site 624:642 the processing site 624:642 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 6 26 from ANGPTL3 1171:1177 arg1 identification 1087:1100 identification 1087:1100 identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3 1087:1177 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 7 27 theme processing 1272:1281 arg1 site 1283:1286 the RAPR(224)↓TT processing site 1255:1286 the RAPR(224)↓TT processing site 1255:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 7 28 theme RAPR 1259:1262 arg1 site 1283:1286 the RAPR(224)↓TT processing site 1255:1286 the RAPR(224)↓TT processing site 1255:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 3 29 dep in 539:540 arg1 vivo 542:545 vivo 542:545 vivo 542:545 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 4 30 theme blood 830:834 arg1 levels 836:841 high triglyceride blood levels 812:841 high triglyceride blood levels 812:841 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 6 31 theme convertase 1141:1150 arg1 processing 1152:1161 the proprotein convertase processing 1126:1161 the proprotein convertase processing 1126:1161 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 0 32 theme convertase 37:46 arg1 activation 48:57 proprotein convertase activation 26:57 proprotein convertase activation of angiopoietin-like protein 3 26:88 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 5 33 gly O-glycosylation 910:924 arg1 proteins 929:936 proteins 929:936 proteins involved in lipid metabolism 929:965 We hypothesized that the GalNAc-T2 transferase performed critical O-glycosylation of proteins involved in lipid metabolism. 20837471 3 34 theme ldlD 567:570 arg1 cells 572:576 CHO ldlD cells 563:576 CHO ldlD cells 563:576 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 4 35 theme GalNAc-transferase 743:760 arg1 gene 791:794 a candidate gene 779:794 a candidate gene for low HDL and high triglyceride blood levels 779:841 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 4 35 theme GalNAc-transferase 743:760 arg1 GALNT2 768:773 GALNT2 768:773 GALNT2 768:773 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 4 35 theme GalNAc-transferase 743:760 arg1 gene 762:765 the polypeptide GalNAc-transferase gene 727:765 the polypeptide GalNAc-transferase gene 727:765 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 2 36 theme processing 421:430 arg1 site 432:435 the processing site 417:435 the processing site 417:435 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 0 37 theme proprotein 26:35 arg1 convertase 37:46 proprotein convertase 26:46 proprotein convertase activation of angiopoietin-like protein 3 26:88 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 1 38 theme promising 308:316 arg1 target 323:328 a promising drug target 306:328 a promising drug target 306:328 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 6 39 gly glycosylated 1054:1065 arg1 sites 1048:1052 potential sites 1038:1052 potential sites glycosylated by GalNAc-T2 1038:1078 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 0 40 from role 100:103 arg1 regulation 144:153 regulation 144:153 regulation of concentrations of plasma lipids 144:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 3 41 theme model 547:551 arg1 system 553:558 an in vivo model system 536:558 an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3 536:672 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 7 42 theme ↓TT 1268:1270 arg1 site 1283:1286 the RAPR(224)↓TT processing site 1255:1286 the RAPR(224)↓TT processing site 1255:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 6 43 dep Thr 1105:1107 arg1 site 1163:1166 adjacent to the proprotein convertase processing site 1114:1166 Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3 1105:1177 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 1 44 theme drug 318:321 arg1 target 323:328 a promising drug target 306:328 a promising drug target 306:328 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 6 45 theme adjacent 1114:1121 arg1 site 1163:1166 adjacent to the proprotein convertase processing site 1114:1166 Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3 1105:1177 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 4 46 theme low 800:802 arg1 HDL 804:806 low HDL 800:806 low HDL 800:806 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 6 47 theme potential 1038:1046 arg1 sites 1048:1052 potential sites 1038:1052 potential sites glycosylated by GalNAc-T2 1038:1078 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 0 48 theme concentrations 158:171 arg1 regulation 144:153 regulation 144:153 regulation of concentrations of plasma lipids 144:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 8 49 theme ANGPTL3 1348:1354 arg1 activation 1356:1365 ANGPTL3 activation 1348:1365 ANGPTL3 activation 1348:1365 The study demonstrates that ANGPTL3 activation is modulated by O-glycosylation and that this step is probably controlled by GalNAc-T2. 20837471 0 50 theme protein 80:86 arg1 activation 48:57 proprotein convertase activation 26:57 proprotein convertase activation of angiopoietin-like protein 3 26:88 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 7 51 dep in 1295:1296 arg1 vitro 1298:1302 vitro 1298:1302 vitro 1298:1302 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 0 52 theme lipids 183:188 arg1 concentrations 158:171 concentrations 158:171 concentrations of plasma lipids 158:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 7 53 gly glycosylation 1211:1223 arg1 Thr 1228:1230 Thr(226) 1228:1235 Thr(226) 1228:1235 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 7 53 gly glycosylation 1211:1223 arg1 peptide 1242:1248 a peptide 1240:1248 a peptide with the RAPR(224)↓TT processing site 1240:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 3 54 theme CHO 563:565 arg1 cells 572:576 CHO ldlD cells 563:576 CHO ldlD cells 563:576 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 4 55 theme association 691:701 arg1 studies 703:709 Genome-wide SNP association studies 675:709 Genome-wide SNP association studies 675:709 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 0 56 theme angiopoietin-like 62:78 arg1 protein 80:86 angiopoietin-like protein 3 62:88 angiopoietin-like protein 3 62:88 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 0 57 theme plasma 176:181 arg1 lipids 183:188 plasma lipids 176:188 plasma lipids 176:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 2 58 theme GalNAc 460:465 arg1 sites 483:487 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 59 gly O-glycosylation 467:481 arg2 sites 483:487 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 59 gly O-glycosylation 467:481 arg2 two 446:448 two 446:448 two 446:448 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 7 60 with peptide 1242:1248 arg1 site 1283:1286 the RAPR(224)↓TT processing site 1255:1286 the RAPR(224)↓TT processing site 1255:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 4 61 theme SNP 687:689 arg1 studies 703:709 Genome-wide SNP association studies 675:709 Genome-wide SNP association studies 675:709 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 7 62 theme furin 1304:1308 arg1 cleavage 1310:1317 in vitro furin cleavage 1295:1317 in vitro furin cleavage 1295:1317 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 7 63 from glycosylation 1211:1223 arg1 peptide 1242:1248 a peptide 1240:1248 a peptide with the RAPR(224)↓TT processing site 1240:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 2 64 theme convertase 360:369 arg1 processing 371:380 proprotein convertase processing 349:380 proprotein convertase processing (RAPR(224)↓TT) for activation 349:410 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 64 theme convertase 360:369 arg1 ↓TT 392:394 RAPR(224)↓TT 383:394 RAPR(224)↓TT 383:394 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 5 65 theme lipid 950:954 arg1 metabolism 956:965 lipid metabolism 950:965 lipid metabolism 950:965 We hypothesized that the GalNAc-T2 transferase performed critical O-glycosylation of proteins involved in lipid metabolism. 20837471 1 66 theme important 239:247 arg1 inhibitor 249:257 an important inhibitor 236:257 an important inhibitor of the endothelial and lipoprotein lipases 236:300 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 1 66 theme important 239:247 arg1 protein 213:219 angiopoietin-like protein 3 195:221 The angiopoietin-like protein 3 (ANGPTL3) 191:231 The angiopoietin-like protein 3 (ANGPTL3) is an important inhibitor of the endothelial and lipoprotein lipases and a promising drug target. 20837471 3 67 theme processing 628:637 arg1 site 639:642 the processing site 624:642 the processing site 624:642 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 2 68 theme proprotein 349:358 arg1 processing 371:380 proprotein convertase processing 349:380 proprotein convertase processing (RAPR(224)↓TT) for activation 349:410 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 2 68 theme proprotein 349:358 arg1 ↓TT 392:394 RAPR(224)↓TT 383:394 RAPR(224)↓TT 383:394 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 3 69 from system 553:558 arg1 cells 572:576 CHO ldlD cells 563:576 CHO ldlD cells 563:576 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 5 70 theme proteins 929:936 arg1 O-glycosylation 910:924 critical O-glycosylation 901:924 critical O-glycosylation of proteins involved in lipid metabolism 901:965 We hypothesized that the GalNAc-T2 transferase performed critical O-glycosylation of proteins involved in lipid metabolism. 20837471 7 71 theme Thr 1228:1230 arg1 glycosylation 1211:1223 GalNAc-T2 glycosylation 1201:1223 GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site 1201:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 20837471 6 72 theme Thr 1105:1107 arg1 identification 1087:1100 identification 1087:1100 identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3 1087:1177 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 0 73 dep modulates 16:24 arg1 role 100:103 possible role 91:103 possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids 91:188 O-glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3: possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids. 20837471 6 74 from identification 1087:1100 arg1 ANGPTL3 1171:1177 ANGPTL3 1171:1177 ANGPTL3 1171:1177 Screening of a panel of proteins known to affect lipid metabolism for potential sites glycosylated by GalNAc-T2 led to identification of Thr(226) adjacent to the proprotein convertase processing site in ANGPTL3. 20837471 3 75 used used 587:590 arg2 system 553:558 an in vivo model system 536:558 an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3 536:672 We developed an in vivo model system in CHO ldlD cells that was used to show that O-glycosylation in the processing site blocked processing of ANGPTL3. 20837471 2 76 theme O-glycosylation 467:481 arg1 sites 483:487 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)) 446:520 ANGPTL3 undergoes proprotein convertase processing (RAPR(224)↓TT) for activation, and the processing site contains two potential GalNAc O-glycosylation sites immediately C-terminal (TT(226)). 20837471 4 77 theme candidate 781:789 arg1 gene 791:794 a candidate gene 779:794 a candidate gene for low HDL and high triglyceride blood levels 779:841 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 4 77 theme candidate 781:789 arg1 gene 762:765 the polypeptide GalNAc-transferase gene 727:765 the polypeptide GalNAc-transferase gene 727:765 Genome-wide SNP association studies have identified the polypeptide GalNAc-transferase gene, GALNT2, as a candidate gene for low HDL and high triglyceride blood levels. 20837471 5 78 theme GalNAc-T2 869:877 arg1 transferase 879:889 the GalNAc-T2 transferase 865:889 the GalNAc-T2 transferase 865:889 We hypothesized that the GalNAc-T2 transferase performed critical O-glycosylation of proteins involved in lipid metabolism. 20837471 7 79 theme GalNAc-T2 1201:1209 arg1 glycosylation 1211:1223 GalNAc-T2 glycosylation 1201:1223 GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site 1201:1286 We demonstrated that GalNAc-T2 glycosylation of Thr(226) in a peptide with the RAPR(224)↓TT processing site blocks in vitro furin cleavage. 21733844 8 0 theme protein 1308:1314 arg1 presence 1270:1277 the presence 1266:1277 the presence of at least one glycosylated protein within each tetramer 1266:1335 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 8 0 theme protein 1308:1314 arg1 sufficient 1340:1349 sufficient 1340:1349 sufficient 1340:1349 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 3 1 from hAQP10 396:401 arg1 pastoris 423:430 the yeast Pichia pastoris 406:430 the yeast Pichia pastoris 406:430 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 7 2 theme stabilizing 1107:1117 arg1 effect 1119:1124 a stabilizing effect 1105:1124 a stabilizing effect of the N-linked glycan 1105:1147 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 4 3 from host 673:676 arg1 overproduced 652:663 overproduced 652:663 overproduced 652:663 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 4 4 theme glycosylation 563:575 arg1 sites 577:581 three predicted glycosylation sites 547:581 three predicted glycosylation sites for hAQP10 547:592 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 7 5 gly nonglycosylated 1069:1083 arg1 protein 1085:1091 the nonglycosylated protein 1065:1091 the nonglycosylated protein 1065:1091 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 1 6 theme glycerol 151:158 arg1 facilitator 121:131 a transmembrane facilitator 105:131 a transmembrane facilitator of both water and glycerol transport in the small intestine 105:191 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 1 6 theme glycerol 151:158 arg1 aquaporin10 81:91 Human aquaporin10 75:91 Human aquaporin10 (hAQP10) 75:100 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 4 7 theme predicted 553:561 arg1 sites 577:581 three predicted glycosylation sites 547:581 three predicted glycosylation sites for hAQP10 547:592 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 6 8 theme glycerol 886:893 arg1 transport 866:874 the transport 862:874 the transport of water, glycerol, erythritol, and xylitol, and glycosylation 862:937 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 2 9 from located 219:225 arg1 membrane 241:248 the apical membrane 230:248 the apical membrane of enterocytes 230:263 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 2 10 located located 219:225 arg2 aquaglyceroporin 199:214 This aquaglyceroporin 194:214 This aquaglyceroporin 194:214 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 2 10 located located 219:225 arg1 membrane 241:248 the apical membrane 230:248 the apical membrane of enterocytes 230:263 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 5 11 theme glycan 727:732 arg1 chromatography 743:756 both glycan affinity chromatography 722:756 chromatography 743:756 Nonglycosylated protein was isolated using both glycan affinity chromatography and through mutating asparagine 133 to a glutamine. 21733844 4 12 from overproduced 652:663 arg1 host 673:676 this host 668:676 this host 668:676 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 1 13 theme Human 75:79 arg1 facilitator 121:131 a transmembrane facilitator 105:131 a transmembrane facilitator of both water and glycerol transport in the small intestine 105:191 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 1 13 theme Human 75:79 arg1 hAQP10 94:99 hAQP10 94:99 hAQP10 94:99 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 1 13 theme Human 75:79 arg1 aquaporin10 81:91 Human aquaporin10 75:91 Human aquaporin10 (hAQP10) 75:100 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 4 14 theme overproduced 652:663 arg1 aquaporins 641:650 the human aquaporins 631:650 the human aquaporins overproduced in this host 631:676 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 4 15 theme human 635:639 arg1 aquaporins 641:650 the human aquaporins 631:650 the human aquaporins overproduced in this host 631:676 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 4 16 theme sites 577:581 arg1 sites 577:581 three predicted glycosylation sites 547:581 three predicted glycosylation sites for hAQP10 547:592 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 4 16 theme sites 577:581 arg1 one 540:542 one 540:542 one 540:542 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 7 17 theme N-linked 1133:1140 arg1 glycan 1142:1147 the N-linked glycan 1129:1147 the N-linked glycan 1129:1147 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 1 18 from facilitator 121:131 arg1 intestine 183:191 the small intestine 173:191 the small intestine 173:191 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 6 19 theme glycosylation 925:937 arg1 transport 866:874 the transport 862:874 the transport of water, glycerol, erythritol, and xylitol, and glycosylation 862:937 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 4 20 gly glycosylation 563:575 arg2 three 547:551 three 547:551 three 547:551 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 4 20 gly glycosylation 563:575 arg2 sites 577:581 three predicted glycosylation sites 547:581 three predicted glycosylation sites for hAQP10 547:592 This finding confirms one of three predicted glycosylation sites for hAQP10, and its glycosylation is unique for the human aquaporins overproduced in this host. 21733844 6 21 theme water 879:883 arg1 transport 866:874 the transport 862:874 the transport of water, glycerol, erythritol, and xylitol, and glycosylation 862:937 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 3 22 theme yeast 410:414 arg1 pastoris 423:430 the yeast Pichia pastoris 406:430 the yeast Pichia pastoris 406:430 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 7 23 theme nonglycosylated 1069:1083 arg1 protein 1085:1091 the nonglycosylated protein 1065:1091 the nonglycosylated protein 1065:1091 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 2 24 from membrane 241:248 arg1 located 219:225 located 219:225 located 219:225 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 1 25 theme small 177:181 arg1 intestine 183:191 the small intestine 173:191 the small intestine 173:191 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 6 26 theme little 943:948 arg1 effect 950:955 little effect 943:955 little effect 943:955 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 5 27 theme Nonglycosylated 679:693 arg1 protein 695:701 Nonglycosylated protein 679:701 Nonglycosylated protein 679:701 Nonglycosylated protein was isolated using both glycan affinity chromatography and through mutating asparagine 133 to a glutamine. 21733844 3 28 from Asn-133 481:487 arg1 C 515:515 the extracellular loop C 492:515 the extracellular loop C 492:515 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 3 29 gly glycosylated 465:476 arg2 Asn-133 481:487 Asn-133 481:487 Asn-133 in the extracellular loop C 481:515 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 3 29 gly glycosylated 465:476 arg1 protein 454:460 the protein 450:460 the protein 450:460 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 8 30 theme hAQP10 1176:1181 arg1 third 1167:1171 third 1167:1171 third 1167:1171 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 8 31 theme tetramer 1435:1442 arg1 protomers 1418:1426 the remaining hAQP10 protomers 1397:1426 the remaining hAQP10 protomers of the tetramer 1397:1442 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 1 32 dep water 141:145 arg1 transport 160:168 transport 160:168 transport 160:168 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 3 33 theme extracellular 496:508 arg1 C 515:515 the extracellular loop C 492:515 the extracellular loop C 492:515 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 0 34 theme human 47:51 arg1 protein 66:72 human aquaporin 10 protein 47:72 human aquaporin 10 protein 47:72 Glycosylation increases the thermostability of human aquaporin 10 protein. 21733844 5 35 gly Nonglycosylated 679:693 arg1 protein 695:701 Nonglycosylated protein 679:701 Nonglycosylated protein 679:701 Nonglycosylated protein was isolated using both glycan affinity chromatography and through mutating asparagine 133 to a glutamine. 21733844 5 36 dep chromatography 743:756 arg1 both 722:725 both 722:725 both 722:725 Nonglycosylated protein was isolated using both glycan affinity chromatography and through mutating asparagine 133 to a glutamine. 21733844 8 37 theme glycosylated 1295:1306 arg1 protein 1308:1314 at least one glycosylated protein 1282:1314 at least one glycosylated protein 1282:1314 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 3 38 theme Pichia 416:421 arg1 pastoris 423:430 the yeast Pichia pastoris 406:430 the yeast Pichia pastoris 406:430 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 7 39 theme glycan 1142:1147 arg1 effect 1119:1124 a stabilizing effect 1105:1124 a stabilizing effect of the N-linked glycan 1105:1147 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 7 40 theme °C 1048:1049 arg1 thermostability 1025:1039 increased thermostability 1015:1039 increased thermostability of 3-6 °C 1015:1049 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 6 41 contain had 939:941 arg2 effect 950:955 little effect 943:955 little effect 943:955 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 6 41 contain had 939:941 arg1 forms 820:824 All three forms 810:824 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation 810:937 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 7 42 theme glycosylated 988:999 arg1 hAQP10 1001:1006 glycosylated hAQP10 988:1006 glycosylated hAQP10 988:1006 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 5 43 theme affinity 734:741 arg1 chromatography 743:756 both glycan affinity chromatography 722:756 chromatography 743:756 Nonglycosylated protein was isolated using both glycan affinity chromatography and through mutating asparagine 133 to a glutamine. 21733844 3 44 theme overproduced 383:394 arg1 hAQP10 396:401 overproduced hAQP10 383:401 overproduced hAQP10 383:401 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 7 45 gly glycosylated 988:999 arg1 hAQP10 1001:1006 glycosylated hAQP10 988:1006 glycosylated hAQP10 988:1006 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 7 46 link N-linked 1133:1140 arg1 glycan 1142:1147 the N-linked glycan 1129:1147 the N-linked glycan 1129:1147 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 2 47 theme glycerol 323:330 arg1 passage 302:308 the passage 298:308 the passage of water and glycerol through these intestinal absorptive cells 298:372 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 0 48 theme aquaporin 53:61 arg1 protein 66:72 human aquaporin 10 protein 47:72 human aquaporin 10 protein 47:72 Glycosylation increases the thermostability of human aquaporin 10 protein. 21733844 8 49 gly glycosylated 1187:1198 arg1 hAQP10 1176:1181 hAQP10 1176:1181 hAQP10 was glycosylated 1176:1198 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 1 50 theme water 141:145 arg1 facilitator 121:131 a transmembrane facilitator 105:131 a transmembrane facilitator of both water and glycerol transport in the small intestine 105:191 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 1 50 theme water 141:145 arg1 aquaporin10 81:91 Human aquaporin10 75:91 Human aquaporin10 (hAQP10) 75:100 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 1 51 theme transmembrane 107:119 arg1 facilitator 121:131 a transmembrane facilitator 105:131 a transmembrane facilitator of both water and glycerol transport in the small intestine 105:191 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 1 51 theme transmembrane 107:119 arg1 aquaporin10 81:91 Human aquaporin10 75:91 Human aquaporin10 (hAQP10) 75:100 Human aquaporin10 (hAQP10) is a transmembrane facilitator of both water and glycerol transport in the small intestine. 21733844 5 52 dep using 716:720 arg1 mutating 770:777 mutating 770:777 mutating asparagine 133 to a glutamine 770:807 Nonglycosylated protein was isolated using both glycan affinity chromatography and through mutating asparagine 133 to a glutamine. 21733844 6 53 theme hAQP10 829:834 arg1 forms 820:824 All three forms 810:824 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation 810:937 All three forms of hAQP10 where found to facilitate the transport of water, glycerol, erythritol, and xylitol, and glycosylation had little effect on functionality. 21733844 2 54 theme enterocytes 253:263 arg1 membrane 241:248 the apical membrane 230:248 the apical membrane of enterocytes 230:263 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 8 55 theme structural 1373:1382 arg1 stability 1384:1392 an enhanced structural stability 1361:1392 an enhanced structural stability to the remaining hAQP10 protomers of the tetramer 1361:1442 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 2 56 theme water 313:317 arg1 passage 302:308 the passage 298:308 the passage of water and glycerol through these intestinal absorptive cells 298:372 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 8 57 theme enhanced 1364:1371 arg1 stability 1384:1392 an enhanced structural stability 1361:1392 an enhanced structural stability to the remaining hAQP10 protomers of the tetramer 1361:1442 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 7 58 theme increased 1015:1023 arg1 thermostability 1025:1039 increased thermostability 1015:1039 increased thermostability of 3-6 °C 1015:1049 In contrast, glycosylated hAQP10 showed increased thermostability of 3-6 °C compared with the nonglycosylated protein, suggesting a stabilizing effect of the N-linked glycan. 21733844 8 59 theme remaining 1401:1409 arg1 protomers 1418:1426 the remaining hAQP10 protomers 1397:1426 the remaining hAQP10 protomers of the tetramer 1397:1442 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 3 60 theme loop 510:513 arg1 C 515:515 the extracellular loop C 492:515 the extracellular loop C 492:515 Here we overproduced hAQP10 in the yeast Pichia pastoris and observed that the protein is glycosylated at Asn-133 in the extracellular loop C. 21733844 8 61 theme thermostability 1208:1222 arg1 mono-modal 1238:1247 mono-modal 1238:1247 mono-modal 1238:1247 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 8 61 theme thermostability 1208:1222 arg1 titration 1224:1232 the thermostability titration 1204:1232 the thermostability titration 1204:1232 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 2 62 theme absorptive 357:366 arg1 cells 368:372 these intestinal absorptive cells 340:372 these intestinal absorptive cells 340:372 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 8 63 gly glycosylated 1295:1306 arg1 protein 1308:1314 at least one glycosylated protein 1282:1314 at least one glycosylated protein 1282:1314 Because only one third of hAQP10 was glycosylated yet the thermostability titration was mono-modal, we suggest that the presence of at least one glycosylated protein within each tetramer is sufficient to convey an enhanced structural stability to the remaining hAQP10 protomers of the tetramer. 21733844 0 64 theme protein 66:72 arg1 thermostability 28:42 the thermostability 24:42 the thermostability of human aquaporin 10 protein 24:72 Glycosylation increases the thermostability of human aquaporin 10 protein. 21733844 2 65 theme apical 234:239 arg1 membrane 241:248 the apical membrane 230:248 the apical membrane of enterocytes 230:263 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 21733844 2 66 theme intestinal 346:355 arg1 cells 368:372 these intestinal absorptive cells 340:372 these intestinal absorptive cells 340:372 This aquaglyceroporin is located in the apical membrane of enterocytes and is believed to contribute to the passage of water and glycerol through these intestinal absorptive cells. 22387313 10 0 with trafficking 1413:1423 arg1 channels 1481:1488 Kv4 channels 1477:1488 Kv4 channels 1477:1488 In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels. 22387313 2 1 theme voltage-gated 352:364 arg1 channels 376:383 voltage-gated potassium channels 352:383 voltage-gated potassium channels of the Kv4 family 352:401 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 7 2 theme expression 920:929 arg1 levels 931:936 The surface expression levels 908:936 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q 908:986 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 1 3 theme serine 246:251 arg1 protease 253:260 the serine protease DPPIV/CD26 242:271 the serine protease DPPIV/CD26 242:271 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 5 4 dep showed 723:728 arg1 Using 679:683 Using 679:683 Using site-directed mutagenesis (N to Q) 679:718 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 5 5 theme site-directed 685:697 arg1 Q 717:717 N to Q 712:717 N to Q 712:717 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 5 5 theme site-directed 685:697 arg1 mutagenesis 699:709 site-directed mutagenesis 685:709 site-directed mutagenesis (N to Q) 685:718 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 3 6 theme N-glycosylation 524:538 arg1 sites 540:544 eight predicted N-glycosylation sites 508:544 eight predicted N-glycosylation sites 508:544 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 5 7 dep Q 717:717 arg1 to 714:715 to 714:715 to 714:715 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 10 8 theme cell 1432:1435 arg1 surface 1437:1443 the cell surface 1428:1443 the cell surface 1428:1443 In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels. 22387313 10 9 theme functional 1449:1458 arg1 interaction 1460:1470 functional interaction 1449:1470 functional interaction with Kv4 channels 1449:1488 In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels. 22387313 10 10 theme DPP10 1407:1411 arg1 trafficking 1413:1423 DPP10 trafficking 1407:1423 DPP10 trafficking to the cell surface 1407:1443 In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels. 22387313 4 11 from role 609:612 arg1 activity 669:676 functional activity 658:676 functional activity 658:676 In this study we investigated the role of N-glycosylation on DPP10 trafficking and functional activity. 22387313 4 11 from role 609:612 arg1 trafficking 642:652 DPP10 trafficking 636:652 DPP10 trafficking 636:652 In this study we investigated the role of N-glycosylation on DPP10 trafficking and functional activity. 22387313 9 12 theme DPP10 1317:1321 arg1 dimerization 1323:1334 DPP10 dimerization 1317:1334 DPP10 dimerization 1317:1334 Interestingly, N257 was crucial for this function and its substitution to glutamine completely blocked DPP10 sorting to the cell surface and prevented DPP10 dimerization. 22387313 6 13 theme DPP10 836:840 arg1 trafficking 842:852 DPP10 trafficking 836:852 DPP10 trafficking to the plasma membrane 836:875 Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. 22387313 3 14 theme predicted 514:522 arg1 sites 540:544 eight predicted N-glycosylation sites 508:544 eight predicted N-glycosylation sites 508:544 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 6 15 gly Glycosylation 777:789 arg2 residues 809:816 these specific residues 794:816 these specific residues 794:816 Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. 22387313 8 16 theme channel 1064:1070 arg1 complex 1072:1078 the Kv4.3/KChIP2a channel complex 1046:1078 the Kv4.3/KChIP2a channel complex 1046:1078 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 7 17 dep substitutions 945:957 arg1 N119Q 965:969 N119Q 965:969 N119Q 965:969 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 7 17 dep substitutions 945:957 arg1 N90Q 959:962 N90Q 959:962 N90Q 959:962 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 7 17 dep substitutions 945:957 arg1 N342Q 982:986 N342Q 982:986 N342Q 982:986 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 7 17 dep substitutions 945:957 arg1 N257Q 972:976 N257Q 972:976 N257Q 972:976 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 7 17 dep substitutions 945:957 arg1 substitutions 945:957 the substitutions 941:957 the substitutions N90Q, N119Q, N257Q and N342Q 941:986 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 3 18 gly N-glycosylation 524:538 arg2 sites 540:544 eight predicted N-glycosylation sites 508:544 eight predicted N-glycosylation sites 508:544 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 3 18 gly N-glycosylation 524:538 arg2 eight 508:512 eight 508:512 eight 508:512 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 2 19 theme family 396:401 arg1 channels 376:383 voltage-gated potassium channels 352:383 voltage-gated potassium channels of the Kv4 family 352:401 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 8 20 from effect 1119:1124 arg1 kinetics 1156:1163 current kinetics 1148:1163 current kinetics 1148:1163 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 8 21 theme Kv4.3/KChIP2a 1050:1062 arg1 complex 1072:1078 the Kv4.3/KChIP2a channel complex 1046:1078 the Kv4.3/KChIP2a channel complex 1046:1078 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 3 22 theme extracellular 553:565 arg1 domain 567:572 the extracellular domain 549:572 the extracellular domain 549:572 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 8 23 with interaction 1029:1039 arg1 complex 1072:1078 the Kv4.3/KChIP2a channel complex 1046:1078 the Kv4.3/KChIP2a channel complex 1046:1078 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 2 24 theme Kv4 392:394 arg1 family 396:401 the Kv4 family 388:401 the Kv4 family 388:401 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 2 25 theme biophysical 449:459 arg1 properties 461:470 biophysical properties 449:470 biophysical properties 449:470 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 3 26 contain containing 497:506 arg2 sites 540:544 eight predicted N-glycosylation sites 508:544 eight predicted N-glycosylation sites 508:544 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 3 26 contain containing 497:506 arg1 DPP10 473:477 DPP10 473:477 DPP10 473:477 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 3 26 contain containing 497:506 arg1 glycoprotein 484:495 a glycoprotein 482:495 a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain 482:572 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 0 27 with interaction 109:119 arg1 channels 130:137 Kv4 channels 126:137 Kv4 channels 126:137 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 10 28 with interaction 1460:1470 arg1 channels 1481:1488 Kv4 channels 1477:1488 Kv4 channels 1477:1488 In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels. 22387313 10 29 theme Kv4 1477:1479 arg1 channels 1481:1488 Kv4 channels 1477:1488 Kv4 channels 1477:1488 In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels. 22387313 0 30 theme mammalian 23:31 arg1 protein 64:70 the mammalian dipeptidyl aminopeptidase-like protein 10 19:73 the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) 19:81 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 0 30 theme mammalian 23:31 arg1 DPP10 76:80 DPP10 76:80 DPP10 76:80 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 0 31 with trafficking 93:103 arg1 channels 130:137 Kv4 channels 126:137 Kv4 channels 126:137 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 2 32 theme mammalian 308:316 arg1 brain 318:322 the mammalian brain 304:322 the mammalian brain 304:322 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 2 33 theme cell 421:424 arg1 expression 434:443 their cell surface expression 415:443 their cell surface expression 415:443 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 2 34 theme surface 426:432 arg1 expression 434:443 their cell surface expression 415:443 their cell surface expression 415:443 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 0 35 theme Kv4 126:128 arg1 channels 130:137 Kv4 channels 126:137 Kv4 channels 126:137 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 8 36 theme current 1148:1154 arg1 kinetics 1156:1163 current kinetics 1148:1163 current kinetics 1148:1163 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 1 37 theme type 199:202 arg1 protein 175:181 The dipeptidyl aminopeptidase-like protein 10 140:184 The dipeptidyl aminopeptidase-like protein 10 (DPP10) 140:192 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 37 theme type 199:202 arg1 homologue 229:237 a type II transmembrane protein homologue 197:237 a type II transmembrane protein homologue 197:237 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 0 38 theme aminopeptidase-like 44:62 arg1 protein 64:70 the mammalian dipeptidyl aminopeptidase-like protein 10 19:73 the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) 19:81 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 0 38 theme aminopeptidase-like 44:62 arg1 DPP10 76:80 DPP10 76:80 DPP10 76:80 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 6 39 from residues 809:816 arg1 Glycosylation 777:789 Glycosylation 777:789 Glycosylation at these specific residues 777:816 Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. 22387313 8 40 theme hastening 1109:1117 arg1 effect 1119:1124 the hastening effect 1105:1124 the hastening effect of wild type DPP10 on current kinetics 1105:1163 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 6 41 theme flow 892:895 arg1 cytometry 897:905 flow cytometry 892:905 flow cytometry 892:905 Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. 22387313 0 42 gly N-glycosylation 0:14 arg1 protein 64:70 the mammalian dipeptidyl aminopeptidase-like protein 10 19:73 the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) 19:81 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 0 42 gly N-glycosylation 0:14 arg1 DPP10 76:80 DPP10 76:80 DPP10 76:80 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 0 43 theme dipeptidyl 33:42 arg1 protein 64:70 the mammalian dipeptidyl aminopeptidase-like protein 10 19:73 the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) 19:81 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 0 43 theme dipeptidyl 33:42 arg1 DPP10 76:80 DPP10 76:80 DPP10 76:80 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 2 44 with complex 339:345 arg1 channels 376:383 voltage-gated potassium channels 352:383 voltage-gated potassium channels of the Kv4 family 352:401 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 1 45 theme transmembrane 207:219 arg1 protein 175:181 The dipeptidyl aminopeptidase-like protein 10 140:184 The dipeptidyl aminopeptidase-like protein 10 (DPP10) 140:192 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 45 theme transmembrane 207:219 arg1 homologue 229:237 a type II transmembrane protein homologue 197:237 a type II transmembrane protein homologue 197:237 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 5 46 theme N 712:712 arg1 Q 717:717 N to Q 712:717 N to Q 712:717 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 5 46 theme N 712:712 arg1 mutagenesis 699:709 site-directed mutagenesis 685:709 site-directed mutagenesis (N to Q) 685:718 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 0 47 theme protein 64:70 arg1 N-glycosylation 0:14 N-glycosylation 0:14 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) 0:81 N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels. 22387313 9 48 theme DPP10 1269:1273 arg1 sorting 1275:1281 DPP10 sorting 1269:1281 DPP10 sorting to the cell surface 1269:1301 Interestingly, N257 was crucial for this function and its substitution to glutamine completely blocked DPP10 sorting to the cell surface and prevented DPP10 dimerization. 22387313 6 49 theme specific 800:807 arg1 residues 809:816 these specific residues 794:816 these specific residues 794:816 Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. 22387313 8 50 theme type 1134:1137 arg1 effect 1119:1124 the hastening effect 1105:1124 the hastening effect of wild type DPP10 on current kinetics 1105:1163 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 5 51 gly N-glycosylation 735:749 arg2 positions 766:774 six positions 762:774 six positions 762:774 Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. 22387313 3 52 gly glycoprotein 484:495 arg1 DPP10 473:477 DPP10 473:477 DPP10 473:477 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 3 52 gly glycoprotein 484:495 arg1 glycoprotein 484:495 a glycoprotein 482:495 a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain 482:572 DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. 22387313 1 53 theme protein 221:227 arg1 protein 175:181 The dipeptidyl aminopeptidase-like protein 10 140:184 The dipeptidyl aminopeptidase-like protein 10 (DPP10) 140:192 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 53 theme protein 221:227 arg1 homologue 229:237 a type II transmembrane protein homologue 197:237 a type II transmembrane protein homologue 197:237 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 8 54 theme wild 1129:1132 arg1 type 1134:1137 wild type DPP10 1129:1143 wild type DPP10 1129:1143 Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. 22387313 6 55 theme plasma 861:866 arg1 membrane 868:875 the plasma membrane 857:875 the plasma membrane 857:875 Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. 22387313 1 56 theme aminopeptidase-like 155:173 arg1 protein 175:181 The dipeptidyl aminopeptidase-like protein 10 140:184 The dipeptidyl aminopeptidase-like protein 10 (DPP10) 140:192 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 56 theme aminopeptidase-like 155:173 arg1 DPP10 187:191 DPP10 187:191 DPP10 187:191 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 56 theme aminopeptidase-like 155:173 arg1 homologue 229:237 a type II transmembrane protein homologue 197:237 a type II transmembrane protein homologue 197:237 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 4 57 theme N-glycosylation 617:631 arg1 role 609:612 the role 605:612 the role of N-glycosylation on DPP10 trafficking and functional activity 605:676 In this study we investigated the role of N-glycosylation on DPP10 trafficking and functional activity. 22387313 4 58 theme functional 658:667 arg1 activity 669:676 functional activity 658:676 functional activity 658:676 In this study we investigated the role of N-glycosylation on DPP10 trafficking and functional activity. 22387313 1 59 theme dipeptidyl 144:153 arg1 protein 175:181 The dipeptidyl aminopeptidase-like protein 10 140:184 The dipeptidyl aminopeptidase-like protein 10 (DPP10) 140:192 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 59 theme dipeptidyl 144:153 arg1 DPP10 187:191 DPP10 187:191 DPP10 187:191 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 1 59 theme dipeptidyl 144:153 arg1 homologue 229:237 a type II transmembrane protein homologue 197:237 a type II transmembrane protein homologue 197:237 The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. 22387313 7 60 theme substitutions 945:957 arg1 levels 931:936 The surface expression levels 908:936 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q 908:986 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 9 61 theme cell 1290:1293 arg1 surface 1295:1301 the cell surface 1286:1301 the cell surface 1286:1301 Interestingly, N257 was crucial for this function and its substitution to glutamine completely blocked DPP10 sorting to the cell surface and prevented DPP10 dimerization. 22387313 2 62 theme potassium 366:374 arg1 channels 376:383 voltage-gated potassium channels 352:383 voltage-gated potassium channels of the Kv4 family 352:401 In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. 22387313 7 63 theme surface 912:918 arg1 levels 931:936 The surface expression levels 908:936 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q 908:986 The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. 22387313 4 64 theme DPP10 636:640 arg1 trafficking 642:652 DPP10 trafficking 636:652 DPP10 trafficking 636:652 In this study we investigated the role of N-glycosylation on DPP10 trafficking and functional activity. 20188224 5 0 theme in-gel 634:639 arg1 deglycosylation 641:655 in-gel deglycosylation 634:655 in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides 634:756 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 12 1 theme structural 1837:1846 arg1 basis 1848:1852 the structural basis 1833:1852 the structural basis of PGRN action 1833:1867 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 4 2 theme oligosaccharides 557:572 arg1 analysis 536:543 detailed analysis 527:543 detailed analysis of the PGRN oligosaccharides and glycopeptides 527:590 A method tailored to enable detailed analysis of the PGRN oligosaccharides and glycopeptides has been developed. 20188224 9 3 theme in-solution 1268:1278 arg1 digestion 1288:1296 in-solution pronase digestion 1268:1296 in-solution pronase digestion 1268:1296 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 6 4 theme sample 796:801 arg1 clean-up 803:810 rapid sample clean-up 790:810 rapid sample clean-up 790:810 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 2 5 theme cancer 259:264 arg1 growth 266:271 cancer growth 259:271 cancer growth 259:271 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 1 6 theme granulin 159:166 arg1 domain 168:173 the granulin domain 155:173 the granulin domain 155:173 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 7 7 from glycans 1047:1053 arg1 run 1078:1080 one chromatographic run 1058:1080 one chromatographic run 1058:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 2 8 theme PGRN 293:296 arg1 mutations 280:288 mutations 280:288 mutations of PGRN 280:296 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 1 9 theme short 188:192 arg1 sequences 201:209 short spacer sequences 188:209 short spacer sequences 188:209 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 3 10 theme glycoprotein 389:400 arg1 PGRN 379:382 PGRN 379:382 PGRN 379:382 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 3 10 theme glycoprotein 389:400 arg1 three 454:458 three 454:458 three 454:458 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 3 10 theme glycoprotein 389:400 arg1 glycoprotein 389:400 a glycoprotein 387:400 a glycoprotein 387:400 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 7 11 from run 1078:1080 arg1 characterization 1009:1024 structural characterization 998:1024 structural characterization of released N-linked glycans in one chromatographic run 998:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 9 12 theme particular 1342:1351 arg1 sites 1353:1357 particular sites 1342:1357 particular sites 1342:1357 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 1 13 theme modular 117:123 arg1 protein 125:131 a modular protein 115:131 a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences 115:209 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 1 13 theme modular 117:123 arg1 PGRN 107:110 PGRN 107:110 PGRN 107:110 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 11 14 theme sites 1589:1593 arg1 Glycosylation 1514:1526 Glycosylation 1514:1526 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites 1514:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 5 15 theme PNGase 688:693 arg1 F 685:685 peptide-N-glycosidase F 663:685 peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides 663:756 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 5 15 theme PNGase 688:693 arg1 F 695:695 PNGase F 688:695 PNGase F 688:695 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 7 16 theme glycans 1047:1053 arg1 characterization 1009:1024 structural characterization 998:1024 structural characterization of released N-linked glycans in one chromatographic run 998:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 12 17 theme important 1792:1800 arg1 implications 1802:1813 important implications 1792:1813 important implications for understanding the structural basis of PGRN action 1792:1867 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 11 18 theme N-glycosylation 1563:1577 arg1 sites 1589:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 2 19 theme PGRN 235:238 arg1 expression 221:230 Elevated expression 212:230 Elevated expression of PGRN 212:238 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 7 20 theme released 1029:1036 arg1 glycans 1047:1053 released N-linked glycans 1029:1053 released N-linked glycans in one chromatographic run 1029:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 12 21 theme granulin 1759:1766 arg1 domains 1768:1774 granulin domains 1759:1774 granulin domains 1759:1774 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 7 22 from characterization 1009:1024 arg1 run 1078:1080 one chromatographic run 1058:1080 one chromatographic run 1058:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 8 23 theme In-gel 1083:1088 arg1 digestion 1098:1106 In-gel tryptic digestion 1083:1106 In-gel tryptic digestion 1083:1106 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 0 24 theme N-glycosylation 90:104 arg1 analysis 66:73 the analysis 62:73 the analysis of progranulin N-glycosylation 62:104 Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation. 20188224 12 25 theme glycosylation 1726:1738 arg1 sites 1740:1744 the observed glycosylation sites 1713:1744 the observed glycosylation sites 1713:1744 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 5 26 theme released 732:739 arg1 oligosaccharides 741:756 the released oligosaccharides 728:756 the released oligosaccharides 728:756 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 7 27 theme oligosaccharides 971:986 arg1 analysis 945:952 analysis 945:952 analysis of permethylated oligosaccharides 945:986 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 8 28 gly deglycosylated 1157:1170 arg1 protein 1172:1178 deglycosylated protein 1157:1178 deglycosylated protein 1157:1178 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 2 29 theme onset 355:359 arg1 form 361:364 an early onset form 346:364 an early onset form of dementia 346:376 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 29 theme onset 355:359 arg1 degeneration 325:336 frontotemporal lobar degeneration 304:336 frontotemporal lobar degeneration (FTLD) 304:343 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 9 30 theme pronase 1280:1286 arg1 digestion 1288:1296 in-solution pronase digestion 1268:1296 in-solution pronase digestion 1268:1296 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 11 31 theme potential 1548:1556 arg1 sites 1589:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 3 32 theme consensus 435:443 arg1 sequons 445:451 five N-glycosylation consensus sequons 414:451 five N-glycosylation consensus sequons 414:451 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 8 33 theme gel 1135:1137 arg1 pieces 1139:1144 the gel pieces 1131:1144 the gel pieces containing deglycosylated protein 1131:1178 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 6 34 theme fragmentation 878:890 arg1 sensitivity 827:837 sensitivity 827:837 sensitivity of MS detection and mass spectrometric fragmentation 827:890 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 4 35 theme detailed 527:534 arg1 analysis 536:543 detailed analysis 527:543 detailed analysis of the PGRN oligosaccharides and glycopeptides 527:590 A method tailored to enable detailed analysis of the PGRN oligosaccharides and glycopeptides has been developed. 20188224 9 36 attach attached 1330:1337 arg2 N-glycan 1321:1328 N-glycan 1321:1328 N-glycan attached at particular sites 1321:1357 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 9 36 attach attached 1330:1337 arg3 sites 1353:1357 particular sites 1342:1357 particular sites 1342:1357 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 0 37 theme mass 31:34 arg1 methods 50:56 mass spectrometric methods 31:56 mass spectrometric methods for the analysis of progranulin N-glycosylation 31:104 Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation. 20188224 6 38 theme mass 859:862 arg1 fragmentation 878:890 mass spectrometric fragmentation 859:890 mass spectrometric fragmentation 859:890 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 7 39 used used 936:939 arg2 LC-ESI-MS/MS 919:930 Reversed-phase monolithic LC-ESI-MS/MS 893:930 Reversed-phase monolithic LC-ESI-MS/MS 893:930 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 3 40 gly N-glycosylation 419:433 arg2 sequons 445:451 five N-glycosylation consensus sequons 414:451 five N-glycosylation consensus sequons 414:451 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 0 41 theme methods 50:56 arg1 application 16:26 application 16:26 application 16:26 Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation. 20188224 0 41 theme methods 50:56 arg1 Development 0:10 Development 0:10 Development 0:10 Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation. 20188224 6 42 theme detection 845:853 arg1 sensitivity 827:837 sensitivity 827:837 sensitivity of MS detection and mass spectrometric fragmentation 827:890 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 10 43 theme glycosylation 1499:1511 arg1 sites 1490:1494 the sites 1486:1494 the sites of glycosylation 1486:1511 The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. 20188224 10 43 theme glycosylation 1499:1511 arg1 glycosylation 1499:1511 glycosylation 1499:1511 glycosylation 1499:1511 The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. 20188224 2 44 theme lobar 319:323 arg1 degeneration 325:336 frontotemporal lobar degeneration 304:336 frontotemporal lobar degeneration (FTLD) 304:343 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 44 theme lobar 319:323 arg1 form 361:364 an early onset form 346:364 an early onset form of dementia 346:376 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 44 theme lobar 319:323 arg1 FTLD 339:342 FTLD 339:342 FTLD 339:342 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 3 45 theme N-glycosylation 419:433 arg1 sequons 445:451 five N-glycosylation consensus sequons 414:451 five N-glycosylation consensus sequons 414:451 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 3 46 gly glycoprotein 389:400 arg1 PGRN 379:382 PGRN 379:382 PGRN 379:382 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 3 46 gly glycoprotein 389:400 arg1 glycoprotein 389:400 a glycoprotein 387:400 a glycoprotein 387:400 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 8 47 contain containing 1146:1155 arg1 pieces 1139:1144 the gel pieces 1131:1144 the gel pieces containing deglycosylated protein 1131:1178 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 8 47 contain containing 1146:1155 arg2 protein 1172:1178 deglycosylated protein 1157:1178 deglycosylated protein 1157:1178 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 8 48 theme site 1201:1204 arg1 determination 1206:1218 N-glycosylation site determination 1185:1218 N-glycosylation site determination 1185:1218 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 7 49 theme Reversed-phase 893:906 arg1 LC-ESI-MS/MS 919:930 Reversed-phase monolithic LC-ESI-MS/MS 893:930 Reversed-phase monolithic LC-ESI-MS/MS 893:930 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 12 50 theme PGRN 1857:1860 arg1 action 1862:1867 PGRN action 1857:1867 PGRN action 1857:1867 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 4 51 theme PGRN 552:555 arg1 oligosaccharides 557:572 the PGRN oligosaccharides 548:572 the PGRN oligosaccharides 548:572 A method tailored to enable detailed analysis of the PGRN oligosaccharides and glycopeptides has been developed. 20188224 10 52 theme glycans 1462:1468 arg1 structures 1435:1444 the structures 1431:1444 the structures of the released glycans 1431:1468 The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. 20188224 9 53 gly glycopeptides 1234:1246 arg2 glycopeptides 1234:1246 glycopeptides 1234:1246 glycopeptides 1234:1246 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 11 54 gly Glycosylation 1514:1526 arg1 sites 1589:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 6 55 theme rapid 790:794 arg1 clean-up 803:810 rapid sample clean-up 790:810 rapid sample clean-up 790:810 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 1 56 theme domain 168:173 arg1 repeats 144:150 7 1/2 repeats 138:150 7 1/2 repeats of the granulin domain separated by short spacer sequences 138:209 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 7 57 link N-linked 1038:1045 arg1 glycans 1047:1053 released N-linked glycans 1029:1053 released N-linked glycans in one chromatographic run 1029:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 9 58 theme N-glycan 1321:1328 arg1 species 1310:1316 species 1310:1316 species of N-glycan attached at particular sites 1310:1357 In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. 20188224 1 59 theme spacer 194:199 arg1 sequences 201:209 short spacer sequences 188:209 short spacer sequences 188:209 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 5 60 theme peptide-N-glycosidase 663:683 arg1 F 685:685 peptide-N-glycosidase F 663:685 peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides 663:756 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 5 60 theme peptide-N-glycosidase 663:683 arg1 F 695:695 PNGase F 688:695 PNGase F 688:695 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 2 61 theme Elevated 212:219 arg1 expression 221:230 Elevated expression 212:230 Elevated expression of PGRN 212:238 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 11 62 theme consensus 1579:1587 arg1 sites 1589:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 11 63 gly N-glycosylation 1563:1577 arg2 five 1543:1546 five 1543:1546 five 1543:1546 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 11 63 gly N-glycosylation 1563:1577 arg2 sites 1589:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 7 64 theme chromatographic 1062:1076 arg1 run 1078:1080 one chromatographic run 1058:1080 one chromatographic run 1058:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 3 65 contain containing 403:412 arg2 sequons 445:451 five N-glycosylation consensus sequons 414:451 five N-glycosylation consensus sequons 414:451 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 3 65 contain containing 403:412 arg1 PGRN 379:382 PGRN 379:382 PGRN 379:382 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 3 65 contain containing 403:412 arg1 glycoprotein 389:400 a glycoprotein 387:400 a glycoprotein 387:400 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 11 66 theme PGRN 1558:1561 arg1 sites 1589:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 four out of five potential PGRN N-glycosylation consensus sites 1531:1593 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 5 67 theme oligosaccharides 741:756 arg1 permethylation 710:723 permethylation 710:723 permethylation of the released oligosaccharides 710:756 The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. 20188224 0 68 theme progranulin 78:88 arg1 N-glycosylation 90:104 progranulin N-glycosylation 78:104 progranulin N-glycosylation 78:104 Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation. 20188224 8 69 theme tryptic 1090:1096 arg1 digestion 1098:1106 In-gel tryptic digestion 1083:1106 In-gel tryptic digestion 1083:1106 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 7 70 theme N-linked 1038:1045 arg1 glycans 1047:1053 released N-linked glycans 1029:1053 released N-linked glycans in one chromatographic run 1029:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 4 71 gly glycopeptides 578:590 arg2 glycopeptides 578:590 glycopeptides 578:590 glycopeptides 578:590 A method tailored to enable detailed analysis of the PGRN oligosaccharides and glycopeptides has been developed. 20188224 1 72 with protein 125:131 arg1 repeats 144:150 7 1/2 repeats 138:150 7 1/2 repeats of the granulin domain separated by short spacer sequences 138:209 PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. 20188224 7 73 theme structural 998:1007 arg1 characterization 1009:1024 structural characterization 998:1024 structural characterization of released N-linked glycans in one chromatographic run 998:1080 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 12 74 theme observed 1717:1724 arg1 sites 1740:1744 the observed glycosylation sites 1713:1744 the observed glycosylation sites 1713:1744 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 12 75 theme sites 1740:1744 arg1 sites 1740:1744 the observed glycosylation sites 1713:1744 the observed glycosylation sites 1713:1744 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 12 75 theme sites 1740:1744 arg1 Two 1706:1708 Two 1706:1708 Two 1706:1708 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 10 76 gly glycosylation 1499:1511 arg2 glycosylation 1499:1511 glycosylation 1499:1511 glycosylation 1499:1511 The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. 20188224 10 76 gly glycosylation 1499:1511 arg2 sites 1490:1494 the sites 1486:1494 the sites of glycosylation 1486:1511 The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. 20188224 2 77 theme early 349:353 arg1 form 361:364 an early onset form 346:364 an early onset form of dementia 346:376 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 77 theme early 349:353 arg1 degeneration 325:336 frontotemporal lobar degeneration 304:336 frontotemporal lobar degeneration (FTLD) 304:343 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 7 78 theme permethylated 957:969 arg1 oligosaccharides 971:986 permethylated oligosaccharides 957:986 permethylated oligosaccharides 957:986 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 20188224 12 79 gly glycosylation 1726:1738 arg2 sites 1740:1744 the observed glycosylation sites 1713:1744 the observed glycosylation sites 1713:1744 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 3 80 theme granulin 481:488 arg1 domains 490:496 granulin domains 481:496 granulin domains 481:496 PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. 20188224 0 81 theme spectrometric 36:48 arg1 methods 50:56 mass spectrometric methods 31:56 mass spectrometric methods for the analysis of progranulin N-glycosylation 31:104 Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation. 20188224 8 82 theme deglycosylated 1157:1170 arg1 protein 1172:1178 deglycosylated protein 1157:1178 deglycosylated protein 1157:1178 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 6 83 theme spectrometric 864:876 arg1 fragmentation 878:890 mass spectrometric fragmentation 859:890 mass spectrometric fragmentation 859:890 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 11 84 theme four 1666:1669 arg1 four 1666:1669 four 1666:1669 four 1666:1669 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 11 84 theme four 1666:1669 arg1 one 1655:1657 one 1655:1657 one 1655:1657 Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. 20188224 2 85 theme dementia 369:376 arg1 form 361:364 an early onset form 346:364 an early onset form of dementia 346:376 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 85 theme dementia 369:376 arg1 degeneration 325:336 frontotemporal lobar degeneration 304:336 frontotemporal lobar degeneration (FTLD) 304:343 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 12 86 contain have 1787:1790 arg1 domains 1768:1774 granulin domains 1759:1774 granulin domains 1759:1774 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 12 86 contain have 1787:1790 arg2 implications 1802:1813 important implications 1792:1813 important implications for understanding the structural basis of PGRN action 1792:1867 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 8 87 theme N-glycosylation 1185:1199 arg1 determination 1206:1218 N-glycosylation site determination 1185:1218 N-glycosylation site determination 1185:1218 In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. 20188224 6 88 theme MS 842:843 arg1 detection 845:853 MS detection 842:853 MS detection 842:853 Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. 20188224 4 89 theme glycopeptides 578:590 arg1 analysis 536:543 detailed analysis 527:543 detailed analysis of the PGRN oligosaccharides and glycopeptides 527:590 A method tailored to enable detailed analysis of the PGRN oligosaccharides and glycopeptides has been developed. 20188224 10 90 theme released 1453:1460 arg1 glycans 1462:1468 the released glycans 1449:1468 the released glycans 1449:1468 The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. 20188224 2 91 theme frontotemporal 304:317 arg1 degeneration 325:336 frontotemporal lobar degeneration 304:336 frontotemporal lobar degeneration (FTLD) 304:343 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 91 theme frontotemporal 304:317 arg1 form 361:364 an early onset form 346:364 an early onset form of dementia 346:376 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 2 91 theme frontotemporal 304:317 arg1 FTLD 339:342 FTLD 339:342 FTLD 339:342 Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. 20188224 12 92 theme action 1862:1867 arg1 basis 1848:1852 the structural basis 1833:1852 the structural basis of PGRN action 1833:1867 Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action. 20188224 7 93 theme monolithic 908:917 arg1 LC-ESI-MS/MS 919:930 Reversed-phase monolithic LC-ESI-MS/MS 893:930 Reversed-phase monolithic LC-ESI-MS/MS 893:930 Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. 17924658 7 0 theme specific 1238:1245 arg1 activity 1247:1254 a specific activity 1236:1254 a specific activity of 237 U/mg 1236:1266 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 4 1 theme hydrolysis 609:618 arg1 product 620:626 a hydrolysis product 607:626 a hydrolysis product 607:626 This intermediate can then react with water to release glutamate as a hydrolysis product or with an amino acid or dipeptide to form a transpeptidation product. 17924658 4 1 theme hydrolysis 609:618 arg1 glutamate 594:602 glutamate 594:602 glutamate 594:602 This intermediate can then react with water to release glutamate as a hydrolysis product or with an amino acid or dipeptide to form a transpeptidation product. 17924658 7 2 theme culture 1114:1120 arg1 purification 1137:1148 subsequent purification 1126:1148 subsequent purification by immobilized metal affinity chromatography 1126:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 7 2 theme culture 1114:1120 arg1 L 1109:1109 1 L 1107:1109 1 L of culture 1107:1120 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 10 3 theme enzyme 1668:1673 arg1 Incubation 1650:1659 Incubation 1650:1659 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis 1650:1757 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 4 theme N-linked 1917:1924 arg1 sites 1940:1944 the seven N-linked glycosylation sites 1907:1944 the seven N-linked glycosylation sites 1907:1944 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 5 theme enzymatic 1709:1717 arg1 digest 1719:1724 enzymatic digest 1709:1724 enzymatic digest 1709:1724 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 4 6 theme transpeptidation 673:688 arg1 product 690:696 a transpeptidation product 671:696 a transpeptidation product 671:696 This intermediate can then react with water to release glutamate as a hydrolysis product or with an amino acid or dipeptide to form a transpeptidation product. 17924658 0 7 theme Kinetic 0:6 arg1 characterization 8:23 characterization 8:23 characterization 8:23 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 1 8 theme glutathione 211:221 arg1 control 235:241 glutathione homeostasis control 211:241 glutathione homeostasis control 211:241 Gamma-glutamyltranspeptidase (GGT) is a heterodimeric enzyme important for glutathione homeostasis control. 17924658 6 9 theme present 895:901 arg1 work 903:906 the present work 891:906 the present work 891:906 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 10 10 theme mass 1731:1734 arg1 analysis 1750:1757 mass spectrometric analysis 1731:1757 mass spectrometric analysis 1731:1757 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 11 theme sites 1940:1944 arg1 four 1899:1902 four 1899:1902 four 1899:1902 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 11 theme sites 1940:1944 arg1 sites 1940:1944 the seven N-linked glycosylation sites 1907:1944 the seven N-linked glycosylation sites 1907:1944 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 5 12 theme mechanism 729:737 arg1 study 716:720 Further detailed study 699:720 Further detailed study of the mechanism underlying these reactions 699:764 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 2 13 theme apoptosis 336:344 arg1 inhibition 346:355 apoptosis inhibition 336:355 apoptosis inhibition 336:355 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 5 14 theme multistep 852:860 arg1 purification 862:873 a multistep purification 850:873 a multistep purification from tissue 850:885 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 8 15 theme min-1 1428:1432 arg1 kcat 1407:1410 a kcat 1405:1410 a kcat of 13.4 x 10(3) min-1 1405:1432 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 8 15 theme min-1 1428:1432 arg1 values 1450:1455 apparent KM values 1438:1455 apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine 1438:1511 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 7 16 theme immobilized 1153:1163 arg1 chromatography 1180:1193 immobilized metal affinity chromatography 1153:1193 immobilized metal affinity chromatography 1153:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 8 17 theme apparent 1438:1445 arg1 values 1450:1455 apparent KM values 1438:1455 apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine 1438:1511 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 5 18 theme detailed 707:714 arg1 study 716:720 Further detailed study 699:720 Further detailed study of the mechanism underlying these reactions 699:764 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 11 19 theme kinetic 1968:1974 arg1 data 1976:1979 These structural and kinetic data 1947:1979 These structural and kinetic data 1947:1979 These structural and kinetic data are discussed with respect to a homology model generated to facilitate visualization. 17924658 6 20 theme 68 1023:1024 arg1 kDa 1026:1028 kDa 1026:1028 kDa 1026:1028 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 0 21 theme glycosylation 65:77 arg1 gamma-glutamyltranspeptidase 106:133 recombinant human gamma-glutamyltranspeptidase 88:133 recombinant human gamma-glutamyltranspeptidase 88:133 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 21 theme glycosylation 65:77 arg1 sites 79:83 glycosylation sites 65:83 glycosylation sites 65:83 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 10 22 theme unambiguous 1778:1788 arg1 identification 1790:1803 the first unambiguous identification 1768:1803 the first unambiguous identification of Thr381 1768:1813 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 22 theme unambiguous 1778:1788 arg1 nucleophile 1834:1844 the active site nucleophile 1818:1844 the active site nucleophile of human gamma-glutamyltranspeptidase 1818:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 5 23 from tissue 880:885 arg1 purification 862:873 a multistep purification 850:873 a multistep purification from tissue 850:885 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 8 24 dep 13.4 1415:1418 arg1 3 1425:1425 3 1425:1425 3 1425:1425 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 7 25 theme enzyme 1222:1227 arg1 mg 1207:1208 1.6 mg 1203:1208 1.6 mg of purified enzyme 1203:1227 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 1 26 theme important 197:205 arg1 enzyme 190:195 a heterodimeric enzyme 174:195 a heterodimeric enzyme important for glutathione homeostasis control 174:241 Gamma-glutamyltranspeptidase (GGT) is a heterodimeric enzyme important for glutathione homeostasis control. 17924658 1 26 theme important 197:205 arg1 Gamma-glutamyltranspeptidase 136:163 Gamma-glutamyltranspeptidase 136:163 Gamma-glutamyltranspeptidase (GGT) 136:169 Gamma-glutamyltranspeptidase (GGT) is a heterodimeric enzyme important for glutathione homeostasis control. 17924658 0 27 theme human 100:104 arg1 gamma-glutamyltranspeptidase 106:133 recombinant human gamma-glutamyltranspeptidase 88:133 recombinant human gamma-glutamyltranspeptidase 88:133 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 9 28 theme GGT-mediated 1518:1529 arg1 hydrolysis 1531:1540 The GGT-mediated hydrolysis 1514:1540 The GGT-mediated hydrolysis of glutathione 1514:1555 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 17924658 6 29 theme Pichia 991:996 arg1 pastoris 998:1005 Pichia pastoris 991:1005 Pichia pastoris 991:1005 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 3 30 theme first 379:383 arg1 step 385:388 the first step 375:388 the first step of its ping-pong mechanism 375:415 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 8 31 theme mM 1492:1493 arg1 kcat 1407:1410 a kcat 1405:1410 a kcat of 13.4 x 10(3) min-1 1405:1432 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 8 31 theme mM 1492:1493 arg1 values 1450:1455 apparent KM values 1438:1455 apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine 1438:1511 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 8 32 theme mass 1379:1382 arg1 spectrometry 1384:1395 mass spectrometry 1379:1395 mass spectrometry 1379:1395 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 4 33 dep release 586:592 arg1 form 666:669 form 666:669 to form a transpeptidation product 663:696 This intermediate can then react with water to release glutamate as a hydrolysis product or with an amino acid or dipeptide to form a transpeptidation product. 17924658 5 34 theme protein 826:832 arg1 quantities 812:821 the low quantities 804:821 the low quantities of protein available after a multistep purification from tissue 804:885 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 3 35 theme ping-pong 397:405 arg1 mechanism 407:415 its ping-pong mechanism 393:415 its ping-pong mechanism 393:415 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 7 36 theme metal 1165:1169 arg1 chromatography 1180:1193 immobilized metal affinity chromatography 1153:1193 immobilized metal affinity chromatography 1153:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 10 37 with Incubation 1650:1659 arg1 analysis 1750:1757 mass spectrometric analysis 1731:1757 mass spectrometric analysis 1731:1757 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 37 with Incubation 1650:1659 arg1 digest 1719:1724 enzymatic digest 1709:1724 enzymatic digest 1709:1724 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 37 with Incubation 1650:1659 arg1 inhibitor 1698:1706 a mechanism-based inhibitor 1680:1706 a mechanism-based inhibitor 1680:1706 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 38 theme active 1822:1827 arg1 identification 1790:1803 the first unambiguous identification 1768:1803 the first unambiguous identification of Thr381 1768:1813 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 38 theme active 1822:1827 arg1 nucleophile 1834:1844 the active site nucleophile 1818:1844 the active site nucleophile of human gamma-glutamyltranspeptidase 1818:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 0 39 dep acylation 51:59 arg1 the 47:49 the 47:49 the 47:49 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 6 40 theme His-tagged 1030:1039 arg1 protein 1041:1047 a 68 kDa His-tagged protein 1021:1047 a 68 kDa His-tagged protein 1021:1047 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 3 41 theme acyl-enzyme 513:523 arg1 intermediate 525:536 an acyl-enzyme intermediate 510:536 an acyl-enzyme intermediate 510:536 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 7 42 theme subsequent 1126:1135 arg1 purification 1137:1148 subsequent purification 1126:1148 subsequent purification by immobilized metal affinity chromatography 1126:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 2 43 theme physiological 280:292 arg1 disease 327:333 Parkinson's disease 315:333 Parkinson's disease 315:333 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 2 43 theme physiological 280:292 arg1 disorders 294:302 many physiological disorders 275:302 many physiological disorders 275:302 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 2 43 theme physiological 280:292 arg1 diabetes 362:369 diabetes 362:369 diabetes 362:369 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 2 43 theme physiological 280:292 arg1 inhibition 346:355 apoptosis inhibition 336:355 apoptosis inhibition 336:355 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 10 44 theme human 1849:1853 arg1 gamma-glutamyltranspeptidase 1855:1882 human gamma-glutamyltranspeptidase 1849:1882 human gamma-glutamyltranspeptidase 1849:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 1 45 theme heterodimeric 176:188 arg1 enzyme 190:195 a heterodimeric enzyme 174:195 a heterodimeric enzyme important for glutathione homeostasis control 174:241 Gamma-glutamyltranspeptidase (GGT) is a heterodimeric enzyme important for glutathione homeostasis control. 17924658 1 45 theme heterodimeric 176:188 arg1 Gamma-glutamyltranspeptidase 136:163 Gamma-glutamyltranspeptidase 136:163 Gamma-glutamyltranspeptidase (GGT) 136:169 Gamma-glutamyltranspeptidase (GGT) is a heterodimeric enzyme important for glutathione homeostasis control. 17924658 10 46 theme mechanism-based 1682:1696 arg1 inhibitor 1698:1706 a mechanism-based inhibitor 1680:1706 a mechanism-based inhibitor 1680:1706 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 4 47 theme amino 639:643 arg1 acid 645:648 an amino acid 636:648 an amino acid 636:648 This intermediate can then react with water to release glutamate as a hydrolysis product or with an amino acid or dipeptide to form a transpeptidation product. 17924658 7 48 theme purified 1213:1220 arg1 enzyme 1222:1227 purified enzyme 1213:1227 purified enzyme 1213:1227 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 10 49 link N-linked 1917:1924 arg1 sites 1940:1944 the seven N-linked glycosylation sites 1907:1944 the seven N-linked glycosylation sites 1907:1944 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 6 50 theme human 921:925 arg1 GGT 927:929 human GGT 921:929 human GGT 921:929 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 9 51 theme KM 1610:1611 arg1 value 1613:1617 a KM value 1608:1617 a KM value of 7.3 microM for glutathione 1608:1647 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 17924658 7 52 from expression 1064:1073 arg1 purification 1137:1148 subsequent purification 1126:1148 subsequent purification by immobilized metal affinity chromatography 1126:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 7 52 from expression 1064:1073 arg1 L 1109:1109 1 L 1107:1109 1 L of culture 1107:1120 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 11 53 theme homology 2013:2020 arg1 model 2022:2026 a homology model 2011:2026 a homology model generated to facilitate visualization 2011:2064 These structural and kinetic data are discussed with respect to a homology model generated to facilitate visualization. 17924658 7 54 from secretion 1079:1087 arg1 purification 1137:1148 subsequent purification 1126:1148 subsequent purification by immobilized metal affinity chromatography 1126:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 7 54 from secretion 1079:1087 arg1 L 1109:1109 1 L 1107:1109 1 L of culture 1107:1120 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 7 55 theme enzyme 1097:1102 arg1 expression 1064:1073 The optimized expression 1050:1073 The optimized expression 1050:1073 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 7 55 theme enzyme 1097:1102 arg1 secretion 1079:1087 secretion 1079:1087 secretion 1079:1087 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 0 56 theme acylation 51:59 arg1 identification 29:42 identification 29:42 identification 29:42 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 56 theme acylation 51:59 arg1 characterization 8:23 characterization 8:23 characterization 8:23 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 9 57 theme microM 1626:1631 arg1 value 1613:1617 a KM value 1608:1617 a KM value of 7.3 microM for glutathione 1608:1647 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 17924658 9 57 theme microM 1626:1631 arg1 kcat 1587:1590 a kcat 1585:1590 a kcat of 53 min-1 1585:1602 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 17924658 10 58 theme spectrometric 1736:1748 arg1 analysis 1750:1757 mass spectrometric analysis 1731:1757 mass spectrometric analysis 1731:1757 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 1 59 theme homeostasis 223:233 arg1 control 235:241 glutathione homeostasis control 211:241 glutathione homeostasis control 211:241 Gamma-glutamyltranspeptidase (GGT) is a heterodimeric enzyme important for glutathione homeostasis control. 17924658 10 60 theme glycosylation 1926:1938 arg1 sites 1940:1944 the seven N-linked glycosylation sites 1907:1944 the seven N-linked glycosylation sites 1907:1944 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 7 61 theme optimized 1054:1062 arg1 expression 1064:1073 The optimized expression 1050:1073 The optimized expression 1050:1073 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 10 62 theme first 1772:1776 arg1 identification 1790:1803 the first unambiguous identification 1768:1803 the first unambiguous identification of Thr381 1768:1813 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 62 theme first 1772:1776 arg1 nucleophile 1834:1844 the active site nucleophile 1818:1844 the active site nucleophile of human gamma-glutamyltranspeptidase 1818:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 6 63 theme pPICZalphaA 951:961 arg1 vector 963:968 the pPICZalphaA vector 947:968 the pPICZalphaA vector 947:968 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 8 64 theme KM 1447:1448 arg1 values 1450:1455 apparent KM values 1438:1455 apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine 1438:1511 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 8 65 theme Kinetic 1269:1275 arg1 parameters 1277:1286 Kinetic parameters 1269:1286 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine 1269:1358 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 5 66 theme Further 699:705 arg1 study 716:720 Further detailed study 699:720 Further detailed study of the mechanism underlying these reactions 699:764 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 10 67 theme Thr381 1808:1813 arg1 identification 1790:1803 the first unambiguous identification 1768:1803 the first unambiguous identification of Thr381 1768:1813 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 67 theme Thr381 1808:1813 arg1 nucleophile 1834:1844 the active site nucleophile 1818:1844 the active site nucleophile of human gamma-glutamyltranspeptidase 1818:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 6 68 theme kDa 1026:1028 arg1 protein 1041:1047 a 68 kDa His-tagged protein 1021:1047 a 68 kDa His-tagged protein 1021:1047 In the present work the gene for human GGT was cloned into the pPICZalphaA vector and transformed into Pichia pastoris to express as a 68 kDa His-tagged protein. 17924658 8 69 theme mM 1465:1466 arg1 kcat 1407:1410 a kcat 1405:1410 a kcat of 13.4 x 10(3) min-1 1405:1432 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 8 69 theme mM 1465:1466 arg1 values 1450:1455 apparent KM values 1438:1455 apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine 1438:1511 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 8 70 theme transpeptidation 1296:1311 arg1 reaction 1313:1320 the transpeptidation reaction 1292:1320 the transpeptidation reaction between glutathione and glycylglycine 1292:1358 Kinetic parameters for the transpeptidation reaction between glutathione and glycylglycine were determined by mass spectrometry, giving a kcat of 13.4 x 10(3) min-1 and apparent KM values of 1.11 mM for glutathione and 8.1 mM for glycylglycine. 17924658 0 71 theme sites 79:83 arg1 identification 29:42 identification 29:42 identification 29:42 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 71 theme sites 79:83 arg1 characterization 8:23 characterization 8:23 characterization 8:23 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 11 72 theme structural 1953:1962 arg1 data 1976:1979 These structural and kinetic data 1947:1979 These structural and kinetic data 1947:1979 These structural and kinetic data are discussed with respect to a homology model generated to facilitate visualization. 17924658 0 73 theme recombinant 88:98 arg1 gamma-glutamyltranspeptidase 106:133 recombinant human gamma-glutamyltranspeptidase 88:133 recombinant human gamma-glutamyltranspeptidase 88:133 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 74 theme gamma-glutamyltranspeptidase 106:133 arg1 gamma-glutamyltranspeptidase 106:133 recombinant human gamma-glutamyltranspeptidase 88:133 recombinant human gamma-glutamyltranspeptidase 88:133 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 74 theme gamma-glutamyltranspeptidase 106:133 arg1 acylation 51:59 acylation 51:59 acylation 51:59 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 74 theme gamma-glutamyltranspeptidase 106:133 arg1 sites 79:83 glycosylation sites 65:83 glycosylation sites 65:83 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 75 gly glycosylation 65:77 arg1 gamma-glutamyltranspeptidase 106:133 recombinant human gamma-glutamyltranspeptidase 88:133 recombinant human gamma-glutamyltranspeptidase 88:133 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 75 gly glycosylation 65:77 arg2 gamma-glutamyltranspeptidase 106:133 recombinant human gamma-glutamyltranspeptidase 88:133 recombinant human gamma-glutamyltranspeptidase 88:133 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 0 75 gly glycosylation 65:77 arg2 sites 79:83 glycosylation sites 65:83 glycosylation sites 65:83 Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human gamma-glutamyltranspeptidase. 17924658 7 76 theme affinity 1171:1178 arg1 chromatography 1180:1193 immobilized metal affinity chromatography 1153:1193 immobilized metal affinity chromatography 1153:1193 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 9 77 theme glutathione 1545:1555 arg1 hydrolysis 1531:1540 The GGT-mediated hydrolysis 1514:1540 The GGT-mediated hydrolysis of glutathione 1514:1555 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 17924658 3 78 dep in 443:444 arg1 vivo 446:449 vivo 446:449 vivo 446:449 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 5 79 theme available 834:842 arg1 quantities 812:821 the low quantities 804:821 the low quantities of protein available after a multistep purification from tissue 804:885 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 10 80 gly glycosylation 1926:1938 arg2 seven 1911:1915 seven 1911:1915 seven 1911:1915 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 80 gly glycosylation 1926:1938 arg2 sites 1940:1944 the seven N-linked glycosylation sites 1907:1944 the seven N-linked glycosylation sites 1907:1944 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 2 81 theme many 275:278 arg1 disease 327:333 Parkinson's disease 315:333 Parkinson's disease 315:333 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 2 81 theme many 275:278 arg1 disorders 294:302 many physiological disorders 275:302 many physiological disorders 275:302 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 2 81 theme many 275:278 arg1 diabetes 362:369 diabetes 362:369 diabetes 362:369 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 2 81 theme many 275:278 arg1 inhibition 346:355 apoptosis inhibition 336:355 apoptosis inhibition 336:355 It has also been implicated in many physiological disorders, including Parkinson's disease, apoptosis inhibition, and diabetes. 17924658 3 82 theme mechanism 407:415 arg1 step 385:388 the first step 375:388 the first step of its ping-pong mechanism 375:415 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 10 83 theme site 1829:1832 arg1 identification 1790:1803 the first unambiguous identification 1768:1803 the first unambiguous identification of Thr381 1768:1813 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 83 theme site 1829:1832 arg1 nucleophile 1834:1844 the active site nucleophile 1818:1844 the active site nucleophile of human gamma-glutamyltranspeptidase 1818:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 3 84 theme in 443:444 arg1 substrate 451:459 its in vivo substrate 439:459 its in vivo substrate 439:459 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 3 84 theme in 443:444 arg1 glutathione 426:436 glutathione 426:436 glutathione 426:436 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 5 85 theme low 808:810 arg1 quantities 812:821 the low quantities 804:821 the low quantities of protein available after a multistep purification from tissue 804:885 Further detailed study of the mechanism underlying these reactions is hindered at least for some GGTs by the low quantities of protein available after a multistep purification from tissue. 17924658 10 86 theme gamma-glutamyltranspeptidase 1855:1882 arg1 identification 1790:1803 the first unambiguous identification 1768:1803 the first unambiguous identification of Thr381 1768:1813 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 10 86 theme gamma-glutamyltranspeptidase 1855:1882 arg1 nucleophile 1834:1844 the active site nucleophile 1818:1844 the active site nucleophile of human gamma-glutamyltranspeptidase 1818:1882 Incubation of the enzyme with a mechanism-based inhibitor, enzymatic digest, and mass spectrometric analysis provided the first unambiguous identification of Thr381 as the active site nucleophile of human gamma-glutamyltranspeptidase, and confirmed four of the seven N-linked glycosylation sites. 17924658 3 87 theme intermediate 525:536 arg1 formation 497:505 formation 497:505 formation of an acyl-enzyme intermediate 497:536 In the first step of its ping-pong mechanism it binds glutathione, its in vivo substrate, and releases cysteinylglycine upon formation of an acyl-enzyme intermediate. 17924658 7 88 theme U/mg 1263:1266 arg1 activity 1247:1254 a specific activity 1236:1254 a specific activity of 237 U/mg 1236:1266 The optimized expression and secretion of this enzyme in 1 L of culture and subsequent purification by immobilized metal affinity chromatography yielded 1.6 mg of purified enzyme having a specific activity of 237 U/mg. 17924658 9 89 theme min-1 1598:1602 arg1 value 1613:1617 a KM value 1608:1617 a KM value of 7.3 microM for glutathione 1608:1647 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 17924658 9 89 theme min-1 1598:1602 arg1 kcat 1587:1590 a kcat 1585:1590 a kcat of 53 min-1 1585:1602 The GGT-mediated hydrolysis of glutathione was also studied, providing a kcat of 53 min-1 and a KM value of 7.3 microM for glutathione. 19508227 0 0 theme endoplasmic 96:106 arg1 reticulum 108:116 the endoplasmic reticulum 92:116 the endoplasmic reticulum 92:116 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 0 1 from sites 54:58 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of tetraspanin Tspan-1 at four distinct sites 0:58 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 1 2 gly N-glycosylation 242:256 arg2 sites 258:262 all four potential N-glycosylation sites 223:262 all four potential N-glycosylation sites 223:262 We showed that Tspan-1, a tetraspanin overexpressed in many human cancers, harbours oligosaccharides at all four potential N-glycosylation sites. 19508227 1 2 gly N-glycosylation 242:256 arg2 four 227:230 four 227:230 four 227:230 We showed that Tspan-1, a tetraspanin overexpressed in many human cancers, harbours oligosaccharides at all four potential N-glycosylation sites. 19508227 3 3 theme correct 448:454 arg1 folding 456:462 correct folding 448:462 correct folding 448:462 Glycosylation seemed to be required for correct folding and subsequent transition through the endoplasmic reticulum. 19508227 2 4 contain contain 377:383 arg2 carbohydrates 393:405 complex carbohydrates 385:405 complex carbohydrates 385:405 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 4 contain contain 377:383 arg1 sites 360:364 two distinct glycosylation sites 333:364 two distinct glycosylation sites 333:364 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 5 theme abundant 274:281 arg1 form 283:286 Its most abundant form 265:286 Its most abundant form 265:286 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 1 6 theme many 174:177 arg1 cancers 185:191 many human cancers 174:191 many human cancers 174:191 We showed that Tspan-1, a tetraspanin overexpressed in many human cancers, harbours oligosaccharides at all four potential N-glycosylation sites. 19508227 1 7 theme human 179:183 arg1 cancers 185:191 many human cancers 174:191 many human cancers 174:191 We showed that Tspan-1, a tetraspanin overexpressed in many human cancers, harbours oligosaccharides at all four potential N-glycosylation sites. 19508227 0 8 theme Tspan-1 29:35 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of tetraspanin Tspan-1 at four distinct sites 0:58 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 2 9 theme sugar 316:320 arg1 chains 322:327 only mannose-rich sugar chains 298:327 only mannose-rich sugar chains 298:327 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 0 10 theme tetraspanin 17:27 arg1 Tspan-1 29:35 tetraspanin Tspan-1 17:35 tetraspanin Tspan-1 17:35 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 2 11 contain contained 288:296 arg2 chains 322:327 only mannose-rich sugar chains 298:327 only mannose-rich sugar chains 298:327 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 11 contain contained 288:296 arg1 form 283:286 Its most abundant form 265:286 Its most abundant form 265:286 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 12 theme mannose-rich 303:314 arg1 chains 322:327 only mannose-rich sugar chains 298:327 only mannose-rich sugar chains 298:327 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 3 13 theme endoplasmic 502:512 arg1 reticulum 514:522 the endoplasmic reticulum 498:522 the endoplasmic reticulum 498:522 Glycosylation seemed to be required for correct folding and subsequent transition through the endoplasmic reticulum. 19508227 0 14 gly Glycosylation 0:12 arg2 sites 54:58 four distinct sites 40:58 four distinct sites 40:58 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 0 14 gly Glycosylation 0:12 arg1 Tspan-1 29:35 tetraspanin Tspan-1 17:35 tetraspanin Tspan-1 17:35 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 2 15 gly glycosylation 346:358 arg2 two 333:335 two 333:335 two 333:335 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 15 gly glycosylation 346:358 arg2 sites 360:364 two distinct glycosylation sites 333:364 two distinct glycosylation sites 333:364 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 0 16 theme distinct 45:52 arg1 sites 54:58 four distinct sites 40:58 four distinct sites 40:58 Glycosylation of tetraspanin Tspan-1 at four distinct sites promotes its transition through the endoplasmic reticulum. 19508227 2 17 theme glycosylation 346:358 arg1 sites 360:364 two distinct glycosylation sites 333:364 two distinct glycosylation sites 333:364 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 18 theme distinct 337:344 arg1 sites 360:364 two distinct glycosylation sites 333:364 two distinct glycosylation sites 333:364 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 2 19 theme complex 385:391 arg1 carbohydrates 393:405 complex carbohydrates 385:405 complex carbohydrates 385:405 Its most abundant form contained only mannose-rich sugar chains but two distinct glycosylation sites could also contain complex carbohydrates. 19508227 3 20 theme subsequent 468:477 arg1 transition 479:488 subsequent transition 468:488 subsequent transition 468:488 Glycosylation seemed to be required for correct folding and subsequent transition through the endoplasmic reticulum. 19508227 1 21 theme potential 232:240 arg1 sites 258:262 all four potential N-glycosylation sites 223:262 all four potential N-glycosylation sites 223:262 We showed that Tspan-1, a tetraspanin overexpressed in many human cancers, harbours oligosaccharides at all four potential N-glycosylation sites. 19508227 1 22 theme N-glycosylation 242:256 arg1 sites 258:262 all four potential N-glycosylation sites 223:262 all four potential N-glycosylation sites 223:262 We showed that Tspan-1, a tetraspanin overexpressed in many human cancers, harbours oligosaccharides at all four potential N-glycosylation sites. 22412906 9 0 from membrane 1482:1489 arg1 line 1595:1598 the INS-1 pancreatic beta cell line 1564:1598 the INS-1 pancreatic beta cell line 1564:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 0 from membrane 1482:1489 arg1 expression 1437:1446 expression 1437:1446 expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line 1437:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 7 1 theme GIP 1256:1258 arg1 receptor 1260:1267 the GIP receptor 1252:1267 the GIP receptor 1252:1267 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 10 2 theme Resonance 1781:1789 arg1 Transfer 1798:1805 Bioluminescence Resonance Energy Transfer 1765:1805 Bioluminescence Resonance Energy Transfer 1765:1805 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 1 3 theme endocrine 242:250 arg1 cells 252:256 gut endocrine cells 238:256 gut endocrine cells 238:256 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 10 4 theme GIP 1628:1630 arg1 mutant 1641:1646 a GIP receptor mutant 1626:1646 a GIP receptor mutant lacking N-glycosylation 1626:1670 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 5 5 theme extracellular 922:934 arg1 N-terminus 936:945 a large extracellular N-terminus 914:945 a large extracellular N-terminus 914:945 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 3 6 from membrane 691:698 arg1 exit 618:621 the exit 614:621 the exit of functional GPCRs from the ER 614:653 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 6 from membrane 691:698 arg1 maintenance 665:675 their maintenance 659:675 their maintenance at the plasma membrane 659:698 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 9 7 theme GIP 1455:1457 arg1 receptor 1459:1466 the GIP receptor 1451:1466 the GIP receptor 1451:1466 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 5 8 theme GIP 882:884 arg1 receptors 896:904 both the GIP and GLP-1 receptors 873:904 receptors 896:904 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 10 9 theme heteromer 1850:1858 arg1 formation 1817:1825 formation 1817:1825 formation of a GIP-GLP1 receptor heteromer 1817:1858 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 5 10 theme GLP-1 890:894 arg1 receptors 896:904 both the GIP and GLP-1 receptors 873:904 receptors 896:904 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 1 11 attach released 224:231 arg2 Polypeptide 166:176 Glucose-dependent Insulinotropic Polypeptide 133:176 Glucose-dependent Insulinotropic Polypeptide (GIP) 133:182 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 11 attach released 224:231 arg2 GLP-1 213:217 GLP-1 213:217 GLP-1 213:217 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 11 attach released 224:231 arg2 Peptide-1 202:210 Glucagon-like Peptide-1 188:210 Glucagon-like Peptide-1 (GLP-1) 188:218 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 11 attach released 224:231 arg1 cells 252:256 gut endocrine cells 238:256 gut endocrine cells 238:256 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 11 attach released 224:231 arg2 GIP 179:181 GIP 179:181 GIP 179:181 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 9 12 gly N-glycosylation 1400:1414 arg1 receptor 1459:1466 the GIP receptor 1451:1466 the GIP receptor 1451:1466 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 10 13 theme GIP-GLP1 1832:1839 arg1 heteromer 1850:1858 a GIP-GLP1 receptor heteromer 1830:1858 a GIP-GLP1 receptor heteromer 1830:1858 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 10 14 theme co-expressed 1686:1697 arg1 receptor 1714:1721 co-expressed wild type GLP1 receptor 1686:1721 co-expressed wild type GLP1 receptor 1686:1721 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 9 15 theme receptor 1459:1466 arg1 expression 1437:1446 expression 1437:1446 expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line 1437:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 5 16 theme many 852:855 arg1 GPCRs 866:870 many family B GPCRs 852:870 many family B GPCRs 852:870 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 9 17 theme glucose-induced 1525:1539 arg1 secretion 1549:1557 glucose-induced insulin secretion 1525:1557 glucose-induced insulin secretion from the INS-1 pancreatic beta cell line 1525:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 3 18 theme GPCRs 637:641 arg1 exit 618:621 the exit 614:621 the exit of functional GPCRs from the ER 614:653 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 18 theme GPCRs 637:641 arg1 maintenance 665:675 their maintenance 659:675 their maintenance at the plasma membrane 659:698 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 9 19 theme secretion 1549:1557 arg1 potentiation 1509:1520 efficient GIP potentiation 1495:1520 efficient GIP potentiation 1495:1520 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 19 theme secretion 1549:1557 arg1 membrane 1482:1489 plasma membrane 1475:1489 plasma membrane 1475:1489 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 5 20 theme multiple 952:959 arg1 sites 971:975 multiple consensus sites 952:975 multiple consensus sites for Asn-linked (N)-glycosylation 952:1008 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 1 21 theme Glucose-dependent 133:149 arg1 GIP 179:181 GIP 179:181 GIP 179:181 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 21 theme Glucose-dependent 133:149 arg1 Polypeptide 166:176 Glucose-dependent Insulinotropic Polypeptide 133:176 Glucose-dependent Insulinotropic Polypeptide (GIP) 133:182 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 0 22 theme expression 49:58 arg1 Regulation 0:9 Regulation 0:9 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 0:108 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 1 23 theme coupled 321:327 arg1 GPCRs 340:344 GPCRs 340:344 GPCRs 340:344 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 23 theme coupled 321:327 arg1 receptors 329:337 their cognate G-protein coupled receptors 297:337 their cognate G-protein coupled receptors (GPCRs) 297:345 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 5 24 theme family 857:862 arg1 GPCRs 866:870 many family B GPCRs 852:870 many family B GPCRs 852:870 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 9 25 theme pancreatic 1574:1583 arg1 line 1595:1598 the INS-1 pancreatic beta cell line 1564:1598 the INS-1 pancreatic beta cell line 1564:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 4 26 theme processes 744:752 arg1 importance 713:722 the importance 709:722 the importance of these regulatory processes 709:752 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 9 27 theme cell 1590:1593 arg1 line 1595:1598 the INS-1 pancreatic beta cell line 1564:1598 the INS-1 pancreatic beta cell line 1564:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 28 from line 1595:1598 arg1 potentiation 1509:1520 efficient GIP potentiation 1495:1520 efficient GIP potentiation 1495:1520 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 28 from line 1595:1598 arg1 membrane 1482:1489 plasma membrane 1475:1489 plasma membrane 1475:1489 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 28 from line 1595:1598 arg1 secretion 1549:1557 glucose-induced insulin secretion 1525:1557 glucose-induced insulin secretion from the INS-1 pancreatic beta cell line 1525:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 4 29 theme GLP-1 804:808 arg1 expression 782:791 functional expression 771:791 functional expression of GIP and GLP-1 receptors 771:818 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 0 30 theme receptor 83:90 arg1 heteromerization 92:107 receptor heteromerization 83:107 receptor heteromerization 83:107 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 5 31 theme N 993:993 arg1 -glycosylation 995:1008 Asn-linked (N)-glycosylation 981:1008 Asn-linked (N)-glycosylation 981:1008 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 7 32 from expression 1183:1192 arg1 parallel 1210:1217 parallel 1210:1217 parallel 1210:1217 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 2 33 theme pleiotropic 396:406 arg1 responses 408:416 tissue-selective pleiotropic responses 379:416 tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells 379:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 2 34 theme secretion 471:479 arg1 augmentation 431:442 augmentation 431:442 augmentation of glucose-induced insulin secretion from pancreatic beta cells 431:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 7 35 from function 1198:1205 arg1 parallel 1210:1217 parallel 1210:1217 parallel 1210:1217 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 4 36 dep GIP 796:798 arg1 receptors 810:818 receptors 810:818 receptors 810:818 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 10 37 theme GLP1 1709:1712 arg1 receptor 1714:1721 co-expressed wild type GLP1 receptor 1686:1721 co-expressed wild type GLP1 receptor 1686:1721 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 2 38 theme glucose-induced 447:461 arg1 secretion 471:479 glucose-induced insulin secretion 447:479 glucose-induced insulin secretion from pancreatic beta cells 447:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 4 39 theme GIP 796:798 arg1 expression 782:791 functional expression 771:791 functional expression of GIP and GLP-1 receptors 771:818 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 0 40 theme GIP 14:16 arg1 Regulation 0:9 Regulation 0:9 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 0:108 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 5 41 theme Asn-linked 981:990 arg1 -glycosylation 995:1008 Asn-linked (N)-glycosylation 981:1008 Asn-linked (N)-glycosylation 981:1008 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 7 42 theme cell 1170:1173 arg1 expression 1183:1192 cell surface expression 1170:1192 cell surface expression 1170:1192 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 0 43 theme GLP1 22:25 arg1 expression 49:58 GLP1 receptor cell surface expression 22:58 GLP1 receptor cell surface expression 22:58 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 2 44 theme pancreatic 486:495 arg1 cells 502:506 pancreatic beta cells 486:506 pancreatic beta cells 486:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 10 45 theme Bioluminescence 1765:1779 arg1 Transfer 1798:1805 Bioluminescence Resonance Energy Transfer 1765:1805 Bioluminescence Resonance Energy Transfer 1765:1805 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 0 46 theme cell 36:39 arg1 expression 49:58 GLP1 receptor cell surface expression 22:58 GLP1 receptor cell surface expression 22:58 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 1 47 theme cognate 303:309 arg1 GPCRs 340:344 GPCRs 340:344 GPCRs 340:344 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 47 theme cognate 303:309 arg1 receptors 329:337 their cognate G-protein coupled receptors 297:337 their cognate G-protein coupled receptors (GPCRs) 297:345 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 6 48 theme ovary 1132:1136 arg1 cells 1138:1142 Chinese hamster ovary cells 1116:1142 Chinese hamster ovary cells 1116:1142 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 6 49 gly glycosylated 1060:1071 arg1 residues 1048:1055 these Asn residues 1038:1055 these Asn residues 1038:1055 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 6 49 gly glycosylated 1060:1071 arg1 each 1030:1033 each 1030:1033 each 1030:1033 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 4 50 from impact 761:766 arg1 expression 782:791 functional expression 771:791 functional expression of GIP and GLP-1 receptors 771:818 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 6 51 theme Chinese 1116:1122 arg1 cells 1138:1142 Chinese hamster ovary cells 1116:1142 Chinese hamster ovary cells 1116:1142 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 3 52 theme co-translational 558:573 arg1 N-glycosylation 509:523 N-glycosylation 509:523 N-glycosylation 509:523 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 52 theme co-translational 558:573 arg1 oligomerization 538:552 receptor oligomerization 529:552 receptor oligomerization 529:552 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 52 theme co-translational 558:573 arg1 processes 575:583 co-translational processes 558:583 co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane 558:698 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 5 53 link Asn-linked 981:990 arg1 -glycosylation 995:1008 Asn-linked (N)-glycosylation 981:1008 Asn-linked (N)-glycosylation 981:1008 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 2 54 from augmentation 431:442 arg1 cells 502:506 pancreatic beta cells 486:506 pancreatic beta cells 486:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 10 55 theme Energy 1791:1796 arg1 Transfer 1798:1805 Bioluminescence Resonance Energy Transfer 1765:1805 Bioluminescence Resonance Energy Transfer 1765:1805 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 1 56 theme gut 238:240 arg1 cells 252:256 gut endocrine cells 238:256 gut endocrine cells 238:256 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 10 57 theme receptor 1632:1639 arg1 mutant 1641:1646 a GIP receptor mutant 1626:1646 a GIP receptor mutant lacking N-glycosylation 1626:1670 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 9 58 theme beta 1585:1588 arg1 line 1595:1598 the INS-1 pancreatic beta cell line 1564:1598 the INS-1 pancreatic beta cell line 1564:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 59 theme plasma 1475:1480 arg1 membrane 1482:1489 plasma membrane 1475:1489 plasma membrane 1475:1489 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 5 60 theme B 864:864 arg1 GPCRs 866:870 many family B GPCRs 852:870 many family B GPCRs 852:870 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 10 61 theme receptor 1841:1848 arg1 heteromer 1850:1858 a GIP-GLP1 receptor heteromer 1830:1858 a GIP-GLP1 receptor heteromer 1830:1858 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 9 62 theme GIP 1505:1507 arg1 potentiation 1509:1520 efficient GIP potentiation 1495:1520 efficient GIP potentiation 1495:1520 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 5 63 theme large 916:920 arg1 N-terminus 936:945 a large extracellular N-terminus 914:945 a large extracellular N-terminus 914:945 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 10 64 theme wild 1699:1702 arg1 receptor 1714:1721 co-expressed wild type GLP1 receptor 1686:1721 co-expressed wild type GLP1 receptor 1686:1721 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 9 65 theme efficient 1495:1503 arg1 potentiation 1509:1520 efficient GIP potentiation 1495:1520 efficient GIP potentiation 1495:1520 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 3 66 theme functional 626:635 arg1 GPCRs 637:641 functional GPCRs 626:641 functional GPCRs 626:641 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 9 67 theme insulin 1541:1547 arg1 secretion 1549:1557 glucose-induced insulin secretion 1525:1557 glucose-induced insulin secretion from the INS-1 pancreatic beta cell line 1525:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 8 68 theme endoplasmic 1377:1387 arg1 reticulum 1389:1397 the endoplasmic reticulum 1373:1397 the endoplasmic reticulum 1373:1397 N-glycosylation mainly lengthens receptor half-life by reducing degradation in the endoplasmic reticulum. 22412906 8 69 theme receptor 1327:1334 arg1 half-life 1336:1344 receptor half-life 1327:1344 receptor half-life 1327:1344 N-glycosylation mainly lengthens receptor half-life by reducing degradation in the endoplasmic reticulum. 22412906 6 70 theme hamster 1124:1130 arg1 cells 1138:1142 Chinese hamster ovary cells 1116:1142 Chinese hamster ovary cells 1116:1142 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 1 71 theme Glucagon-like 188:200 arg1 Peptide-1 202:210 Glucagon-like Peptide-1 188:210 Glucagon-like Peptide-1 (GLP-1) 188:218 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 71 theme Glucagon-like 188:200 arg1 GLP-1 213:217 GLP-1 213:217 GLP-1 213:217 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 9 72 theme INS-1 1568:1572 arg1 line 1595:1598 the INS-1 pancreatic beta cell line 1564:1598 the INS-1 pancreatic beta cell line 1564:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 10 73 theme mutant 1641:1646 arg1 expression 1612:1621 Functional expression 1601:1621 Functional expression of a GIP receptor mutant lacking N-glycosylation 1601:1670 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 5 74 contain possess 906:912 arg1 receptors 896:904 both the GIP and GLP-1 receptors 873:904 receptors 896:904 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 5 74 contain possess 906:912 arg2 N-terminus 936:945 a large extracellular N-terminus 914:945 a large extracellular N-terminus 914:945 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 7 75 theme stronger 1230:1237 arg1 control 1239:1245 stronger control 1230:1245 stronger control over the GIP receptor than the GLP-1 receptor 1230:1291 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 1 76 theme Insulinotropic 151:164 arg1 GIP 179:181 GIP 179:181 GIP 179:181 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 76 theme Insulinotropic 151:164 arg1 Polypeptide 166:176 Glucose-dependent Insulinotropic Polypeptide 133:176 Glucose-dependent Insulinotropic Polypeptide (GIP) 133:182 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 4 77 theme regulatory 733:742 arg1 processes 744:752 these regulatory processes 727:752 these regulatory processes 727:752 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 1 78 dep meal 127:130 arg1 response 113:120 response 113:120 response 113:120 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 3 79 from ER 652:653 arg1 exit 618:621 the exit 614:621 the exit of functional GPCRs from the ER 614:653 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 79 from ER 652:653 arg1 maintenance 665:675 their maintenance 659:675 their maintenance at the plasma membrane 659:698 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 6 80 theme human 1085:1089 arg1 receptor 1091:1098 either human receptor 1078:1098 either human receptor 1078:1098 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 2 81 from cells 502:506 arg1 secretion 471:479 glucose-induced insulin secretion 447:479 glucose-induced insulin secretion from pancreatic beta cells 447:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 2 81 from cells 502:506 arg1 augmentation 431:442 augmentation 431:442 augmentation of glucose-induced insulin secretion from pancreatic beta cells 431:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 2 82 theme tissue-selective 379:394 arg1 responses 408:416 tissue-selective pleiotropic responses 379:416 tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells 379:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 5 83 theme consensus 961:969 arg1 sites 971:975 multiple consensus sites 952:975 multiple consensus sites for Asn-linked (N)-glycosylation 952:1008 Like many family B GPCRs, both the GIP and GLP-1 receptors possess a large extracellular N-terminus with multiple consensus sites for Asn-linked (N)-glycosylation. 22412906 2 84 theme insulin 463:469 arg1 secretion 471:479 glucose-induced insulin secretion 447:479 glucose-induced insulin secretion from pancreatic beta cells 447:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 4 85 theme functional 771:780 arg1 expression 782:791 functional expression 771:791 functional expression of GIP and GLP-1 receptors 771:818 Despite the importance of these regulatory processes, their impact on functional expression of GIP and GLP-1 receptors has not been well studied. 22412906 10 86 theme type 1704:1707 arg1 receptor 1714:1721 co-expressed wild type GLP1 receptor 1686:1721 co-expressed wild type GLP1 receptor 1686:1721 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 7 87 theme surface 1175:1181 arg1 expression 1183:1192 cell surface expression 1170:1192 cell surface expression 1170:1192 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 3 88 theme plasma 684:689 arg1 membrane 691:698 the plasma membrane 680:698 the plasma membrane 680:698 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 0 89 theme receptor 27:34 arg1 expression 49:58 GLP1 receptor cell surface expression 22:58 GLP1 receptor cell surface expression 22:58 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 6 90 theme Asn 1044:1046 arg1 residues 1048:1055 these Asn residues 1038:1055 these Asn residues 1038:1055 Here, we show that each of these Asn residues is glycosylated when either human receptor is expressed in Chinese hamster ovary cells. 22412906 2 91 theme beta 497:500 arg1 cells 502:506 pancreatic beta cells 486:506 pancreatic beta cells 486:506 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 22412906 0 92 theme surface 41:47 arg1 expression 49:58 GLP1 receptor cell surface expression 22:58 GLP1 receptor cell surface expression 22:58 Regulation of GIP and GLP1 receptor cell surface expression by N-glycosylation and receptor heteromerization. 22412906 9 93 dep membrane 1482:1489 arg1 the 1471:1473 the 1471:1473 the 1471:1473 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 3 94 theme receptor 529:536 arg1 N-glycosylation 509:523 N-glycosylation 509:523 N-glycosylation 509:523 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 94 theme receptor 529:536 arg1 oligomerization 538:552 receptor oligomerization 529:552 receptor oligomerization 529:552 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 3 94 theme receptor 529:536 arg1 processes 575:583 co-translational processes 558:583 co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane 558:698 N-glycosylation and receptor oligomerization are co-translational processes that are thought to regulate the exit of functional GPCRs from the ER and their maintenance at the plasma membrane. 22412906 1 95 theme G-protein 311:319 arg1 GPCRs 340:344 GPCRs 340:344 GPCRs 340:344 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 1 95 theme G-protein 311:319 arg1 receptors 329:337 their cognate G-protein coupled receptors 297:337 their cognate G-protein coupled receptors (GPCRs) 297:345 In response to a meal, Glucose-dependent Insulinotropic Polypeptide (GIP) and Glucagon-like Peptide-1 (GLP-1) are released from gut endocrine cells into the circulation and interact with their cognate G-protein coupled receptors (GPCRs). 22412906 9 96 from potentiation 1509:1520 arg1 line 1595:1598 the INS-1 pancreatic beta cell line 1564:1598 the INS-1 pancreatic beta cell line 1564:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 9 96 from potentiation 1509:1520 arg1 expression 1437:1446 expression 1437:1446 expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line 1437:1598 N-glycosylation is also required for expression of the GIP receptor at the plasma membrane and efficient GIP potentiation of glucose-induced insulin secretion from the INS-1 pancreatic beta cell line. 22412906 7 97 theme GLP-1 1278:1282 arg1 receptor 1284:1291 the GLP-1 receptor 1274:1291 the GLP-1 receptor 1274:1291 N-glycosylation enhances cell surface expression and function in parallel but exerts stronger control over the GIP receptor than the GLP-1 receptor. 22412906 10 98 theme Functional 1601:1610 arg1 expression 1612:1621 Functional expression 1601:1621 Functional expression of a GIP receptor mutant lacking N-glycosylation 1601:1670 Functional expression of a GIP receptor mutant lacking N-glycosylation is rescued by co-expressed wild type GLP1 receptor, which, together with data obtained using Bioluminescence Resonance Energy Transfer, suggests formation of a GIP-GLP1 receptor heteromer. 22412906 2 99 theme Receptor 348:355 arg1 activation 357:366 Receptor activation 348:366 Receptor activation 348:366 Receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. 17715132 5 0 theme membrane 764:771 arg1 targeting 773:781 plasma membrane targeting 757:781 plasma membrane targeting 757:781 We show for the first time that Pannexin1 is glycosylated at Asn-254 and that this residue is important for plasma membrane targeting. 17715132 5 1 gly glycosylated 694:705 arg2 Asn-254 710:716 Asn-254 710:716 Asn-254 710:716 We show for the first time that Pannexin1 is glycosylated at Asn-254 and that this residue is important for plasma membrane targeting. 17715132 5 1 gly glycosylated 694:705 arg1 Pannexin1 681:689 Pannexin1 681:689 Pannexin1 681:689 We show for the first time that Pannexin1 is glycosylated at Asn-254 and that this residue is important for plasma membrane targeting. 17715132 4 2 dep analyzed 531:538 arg1 Using 496:500 Using 496:500 Using site-directed mutagenesis 496:526 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 6 3 theme extracellular 822:834 arg1 surface 836:842 its extracellular surface 818:842 its extracellular surface 818:842 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 2 4 theme junction-like 345:357 arg1 structures 359:368 gap junction-like structures 341:368 gap junction-like structures 341:368 Based on membrane topology, folding and secondary structure prediction, pannexins are proposed to form gap junction-like structures. 17715132 4 5 link N-linked 555:562 arg1 sites 578:582 three putative N-linked glycosylation sites 540:582 three putative N-linked glycosylation sites 540:582 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 5 6 theme first 665:669 arg1 time 671:674 the first time 661:674 the first time 661:674 We show for the first time that Pannexin1 is glycosylated at Asn-254 and that this residue is important for plasma membrane targeting. 17715132 7 7 theme junctional 999:1008 arg1 areas 1010:1014 Pannexin1 junctional areas 989:1014 Pannexin1 junctional areas 989:1014 Ultrastructural analysis by electron microscopy confirmed that Pannexin1 junctional areas do not appear as canonical gap junctions. 17715132 2 8 theme structure 288:296 arg1 prediction 298:307 secondary structure prediction 278:307 secondary structure prediction 278:307 Based on membrane topology, folding and secondary structure prediction, pannexins are proposed to form gap junction-like structures. 17715132 0 9 theme plasma 80:85 arg1 membrane 87:94 the plasma membrane 76:94 the plasma membrane 76:94 Pannexin1 channels contain a glycosylation site that targets the hexamer to the plasma membrane. 17715132 9 10 theme Pannexin1 1168:1176 arg1 mechanism 1201:1209 a significant mechanism 1187:1209 a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues 1187:1308 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 9 10 theme Pannexin1 1168:1176 arg1 N-glycosylation 1149:1163 N-glycosylation 1149:1163 N-glycosylation of Pannexin1 1149:1176 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 2 11 theme secondary 278:286 arg1 prediction 298:307 secondary structure prediction 278:307 secondary structure prediction 278:307 Based on membrane topology, folding and secondary structure prediction, pannexins are proposed to form gap junction-like structures. 17715132 6 12 theme Pannexin1 805:813 arg1 glycosylation 788:800 The glycosylation 784:800 The glycosylation of Pannexin1 at its extracellular surface 784:842 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 1 13 theme different 163:171 arg1 tissues 173:179 many different tissues 158:179 many different tissues 158:179 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 9 14 theme membrane 1251:1258 arg1 proteins 1260:1267 these membrane proteins 1245:1267 these membrane proteins 1245:1267 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 0 15 contain contain 19:25 arg2 site 43:46 a glycosylation site 27:46 a glycosylation site that targets the hexamer to the plasma membrane 27:94 Pannexin1 channels contain a glycosylation site that targets the hexamer to the plasma membrane. 17715132 0 15 contain contain 19:25 arg1 channels 10:17 Pannexin1 channels 0:17 Pannexin1 channels 0:17 Pannexin1 channels contain a glycosylation site that targets the hexamer to the plasma membrane. 17715132 3 16 theme cell 441:444 arg1 surface 446:452 the cell surface 437:452 the cell surface 437:452 We show here that Pannexin1 forms a hexameric channel and reaches the cell surface but, unlike connexins, is N-glycosylated. 17715132 6 17 gly glycosylation 788:800 arg2 surface 836:842 its extracellular surface 818:842 its extracellular surface 818:842 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 6 17 gly glycosylation 788:800 arg1 surface 836:842 its extracellular surface 818:842 its extracellular surface 818:842 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 6 17 gly glycosylation 788:800 arg1 Pannexin1 805:813 Pannexin1 805:813 Pannexin1 805:813 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 3 18 gly N-glycosylated 480:493 arg1 Pannexin1 389:397 Pannexin1 389:397 Pannexin1 389:397 We show here that Pannexin1 forms a hexameric channel and reaches the cell surface but, unlike connexins, is N-glycosylated. 17715132 0 19 theme Pannexin1 0:8 arg1 channels 10:17 Pannexin1 channels 0:17 Pannexin1 channels 0:17 Pannexin1 channels contain a glycosylation site that targets the hexamer to the plasma membrane. 17715132 4 20 theme glycosylation 564:576 arg1 sites 578:582 three putative N-linked glycosylation sites 540:582 three putative N-linked glycosylation sites 540:582 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 9 21 theme proteins 1260:1267 arg1 trafficking 1230:1240 the trafficking 1226:1240 the trafficking of these membrane proteins to the cell surface in different tissues 1226:1308 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 4 22 theme N-linked 555:562 arg1 sites 578:582 three putative N-linked glycosylation sites 540:582 three putative N-linked glycosylation sites 540:582 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 8 23 theme plasma 1116:1121 arg1 membrane 1123:1130 the plasma membrane 1112:1130 the plasma membrane 1112:1130 Rather, Pannexin1 channels are distributed throughout the plasma membrane. 17715132 7 24 theme Pannexin1 989:997 arg1 areas 1010:1014 Pannexin1 junctional areas 989:1014 Pannexin1 junctional areas 989:1014 Ultrastructural analysis by electron microscopy confirmed that Pannexin1 junctional areas do not appear as canonical gap junctions. 17715132 6 25 theme intercellular 903:915 arg1 channel 917:923 an intercellular channel 900:923 an intercellular channel 900:923 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 1 26 theme many 158:161 arg1 tissues 173:179 many different tissues 158:179 many different tissues 158:179 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 3 27 theme hexameric 407:415 arg1 channel 417:423 a hexameric channel 405:423 a hexameric channel 405:423 We show here that Pannexin1 forms a hexameric channel and reaches the cell surface but, unlike connexins, is N-glycosylated. 17715132 9 28 theme cell 1276:1279 arg1 surface 1281:1287 the cell surface 1272:1287 the cell surface 1272:1287 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 1 29 theme vertebrate 203:212 arg1 system 230:235 the vertebrate central nervous system 199:235 the vertebrate central nervous system 199:235 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 0 30 theme glycosylation 29:41 arg1 site 43:46 a glycosylation site 27:46 a glycosylation site that targets the hexamer to the plasma membrane 27:94 Pannexin1 channels contain a glycosylation site that targets the hexamer to the plasma membrane. 17715132 6 31 from surface 836:842 arg1 glycosylation 788:800 The glycosylation 784:800 The glycosylation of Pannexin1 at its extracellular surface 784:842 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 7 32 theme gap 1043:1045 arg1 junctions 1047:1055 canonical gap junctions 1033:1055 canonical gap junctions 1033:1055 Ultrastructural analysis by electron microscopy confirmed that Pannexin1 junctional areas do not appear as canonical gap junctions. 17715132 1 33 theme central 214:220 arg1 system 230:235 the vertebrate central nervous system 199:235 the vertebrate central nervous system 199:235 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 4 34 gly glycosylation 564:576 arg2 sites 578:582 three putative N-linked glycosylation sites 540:582 three putative N-linked glycosylation sites 540:582 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 4 34 gly glycosylation 564:576 arg2 three 540:544 three 540:544 three 540:544 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 2 35 theme gap 341:343 arg1 structures 359:368 gap junction-like structures 341:368 gap junction-like structures 341:368 Based on membrane topology, folding and secondary structure prediction, pannexins are proposed to form gap junction-like structures. 17715132 4 36 theme site-directed 502:514 arg1 mutagenesis 516:526 site-directed mutagenesis 502:526 site-directed mutagenesis 502:526 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 4 37 theme putative 546:553 arg1 sites 578:582 three putative N-linked glycosylation sites 540:582 three putative N-linked glycosylation sites 540:582 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 9 38 theme different 1292:1300 arg1 tissues 1302:1308 different tissues 1292:1308 different tissues 1292:1308 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 8 39 theme Pannexin1 1066:1074 arg1 channels 1076:1083 Pannexin1 channels 1066:1083 Pannexin1 channels 1066:1083 Rather, Pannexin1 channels are distributed throughout the plasma membrane. 17715132 9 40 gly N-glycosylation 1149:1163 arg1 Pannexin1 1168:1176 Pannexin1 1168:1176 Pannexin1 1168:1176 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 7 41 theme electron 954:961 arg1 microscopy 963:972 electron microscopy 954:972 electron microscopy 954:972 Ultrastructural analysis by electron microscopy confirmed that Pannexin1 junctional areas do not appear as canonical gap junctions. 17715132 0 42 gly glycosylation 29:41 arg2 site 43:46 a glycosylation site 27:46 a glycosylation site that targets the hexamer to the plasma membrane 27:94 Pannexin1 channels contain a glycosylation site that targets the hexamer to the plasma membrane. 17715132 7 43 theme canonical 1033:1041 arg1 junctions 1047:1055 canonical gap junctions 1033:1055 canonical gap junctions 1033:1055 Ultrastructural analysis by electron microscopy confirmed that Pannexin1 junctional areas do not appear as canonical gap junctions. 17715132 1 44 theme discovered 117:126 arg1 proteins 136:143 newly discovered channel proteins 111:143 newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system 111:235 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 1 44 theme discovered 117:126 arg1 Pannexins 97:105 Pannexins 97:105 Pannexins 97:105 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 7 45 theme Ultrastructural 926:940 arg1 analysis 942:949 Ultrastructural analysis 926:949 Ultrastructural analysis by electron microscopy 926:972 Ultrastructural analysis by electron microscopy confirmed that Pannexin1 junctional areas do not appear as canonical gap junctions. 17715132 1 46 theme nervous 222:228 arg1 system 230:235 the vertebrate central nervous system 199:235 the vertebrate central nervous system 199:235 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 1 47 theme channel 128:134 arg1 proteins 136:143 newly discovered channel proteins 111:143 newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system 111:235 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 1 47 theme channel 128:134 arg1 Pannexins 97:105 Pannexins 97:105 Pannexins 97:105 Pannexins are newly discovered channel proteins expressed in many different tissues and abundantly in the vertebrate central nervous system. 17715132 6 48 dep dock 887:890 arg1 unlikely 853:860 unlikely 853:860 unlikely 853:860 The glycosylation of Pannexin1 at its extracellular surface makes it unlikely that two oligomers could dock to form an intercellular channel. 17715132 2 49 theme membrane 247:254 arg1 topology 256:263 membrane topology 247:263 membrane topology 247:263 Based on membrane topology, folding and secondary structure prediction, pannexins are proposed to form gap junction-like structures. 17715132 9 50 theme significant 1189:1199 arg1 mechanism 1201:1209 a significant mechanism 1187:1209 a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues 1187:1308 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 9 50 theme significant 1189:1199 arg1 N-glycosylation 1149:1163 N-glycosylation 1149:1163 N-glycosylation of Pannexin1 1149:1176 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 9 51 from trafficking 1230:1240 arg1 tissues 1302:1308 different tissues 1292:1308 different tissues 1292:1308 We propose that N-glycosylation of Pannexin1 could be a significant mechanism for regulating the trafficking of these membrane proteins to the cell surface in different tissues. 17715132 4 52 theme channel 629:635 arg1 expression 637:646 channel expression 629:646 channel expression 629:646 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 5 53 theme plasma 757:762 arg1 targeting 773:781 plasma membrane targeting 757:781 plasma membrane targeting 757:781 We show for the first time that Pannexin1 is glycosylated at Asn-254 and that this residue is important for plasma membrane targeting. 17715132 4 54 theme mutation 617:624 arg1 effects 601:607 the effects 597:607 the effects of each mutation on channel expression 597:646 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 17715132 4 55 from effects 601:607 arg1 expression 637:646 channel expression 629:646 channel expression 629:646 Using site-directed mutagenesis we analyzed three putative N-linked glycosylation sites and examined the effects of each mutation on channel expression. 20507882 7 0 theme alpha-dystroglycan 1436:1453 arg1 O-glycosylation 1417:1431 the complex in vivo O-glycosylation 1397:1431 the complex in vivo O-glycosylation of alpha-dystroglycan 1397:1453 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 6 1 theme glycan 1058:1063 arg1 sites 1076:1080 glycan attachment sites 1058:1080 glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1058:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 1 theme glycan 1058:1063 arg1 structure 1109:1117 the NeuAcHexHexNAcHex-O structure 1085:1117 the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1085:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 1 2 theme alpha-dystroglycan 180:197 arg1 important 202:210 important 202:210 important 202:210 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 1 2 theme alpha-dystroglycan 180:197 arg1 glycosylation 137:149 The glycosylation 133:149 The glycosylation of the extracellular protein alpha-dystroglycan 133:197 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 0 3 theme alpha-dystroglycan 86:103 arg1 alpha-dystroglycan 86:103 alpha-dystroglycan 86:103 alpha-dystroglycan 86:103 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 0 3 theme alpha-dystroglycan 86:103 arg1 domain 76:81 the mucin-like domain 61:81 the mucin-like domain of alpha-dystroglycan from human skeletal muscle 61:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 5 4 gly glycosylation 994:1006 arg2 site 1008:1011 glycosylation site occupancy 994:1021 glycosylation site occupancy 994:1021 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 5 4 gly glycosylation 994:1006 arg2 occupancy 1013:1021 glycosylation site occupancy 994:1021 glycosylation site occupancy 994:1021 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 0 5 theme skeletal 116:123 arg1 muscle 125:130 human skeletal muscle 110:130 human skeletal muscle 110:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 6 6 theme first 1047:1051 arg1 time 1053:1056 the first time 1043:1056 the first time 1043:1056 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 7 7 gly glycopeptides 1308:1320 arg2 glycopeptides 1308:1320 Twenty-five glycopeptides 1296:1320 Twenty-five glycopeptides 1296:1320 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 1 8 theme several 301:307 arg1 forms 309:313 several forms 301:313 several forms of congenital muscular dystrophies 301:348 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 0 9 theme human 110:114 arg1 muscle 125:130 human skeletal muscle 110:130 human skeletal muscle 110:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 7 10 gly O-glycosylation 1417:1431 arg1 alpha-dystroglycan 1436:1453 alpha-dystroglycan 1436:1453 alpha-dystroglycan 1436:1453 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 3 11 theme C-terminal 658:667 arg1 alpha-dystroglycan 679:696 alpha-dystroglycan 679:696 alpha-dystroglycan 679:696 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 3 11 theme C-terminal 658:667 arg1 domain 669:674 the mucin-like and C-terminal domain 639:674 domain 669:674 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 2 12 dep immunoprecipitation 354:372 arg1 strategies 421:430 capture-and-release enrichment strategies 390:430 capture-and-release enrichment strategies 390:430 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 7 13 theme in 1409:1410 arg1 O-glycosylation 1417:1431 the complex in vivo O-glycosylation 1397:1431 the complex in vivo O-glycosylation of alpha-dystroglycan 1397:1453 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 7 14 theme Twenty-five 1296:1306 arg1 glycopeptides 1308:1320 Twenty-five glycopeptides 1296:1320 Twenty-five glycopeptides 1296:1320 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 4 15 theme anchored 760:767 arg1 structures 769:778 anchored structures 760:778 anchored structures 760:778 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 2 16 theme alpha-dystroglycan 470:487 arg1 glycopeptides 453:465 tryptic glycopeptides 445:465 tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle 445:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 6 17 theme anticipated 1140:1150 arg1 sLacNAc-Man 1197:1207 sLacNAc-Man 1197:1207 sLacNAc-Man 1197:1207 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 17 theme anticipated 1140:1150 arg1 Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan 1152:1194 the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan 1136:1194 the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1136:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 3 18 theme Nano-liquid 517:527 arg1 spectrometry 556:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 6 19 from domain 1233:1238 arg1 muscle 1288:1293 human skeletal muscle 1273:1293 human skeletal muscle 1273:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 3 20 theme mucin-like 643:652 arg1 alpha-dystroglycan 679:696 alpha-dystroglycan 679:696 alpha-dystroglycan 679:696 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 3 20 theme mucin-like 643:652 arg1 domain 669:674 the mucin-like and C-terminal domain 639:674 domain 669:674 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 3 21 theme tandem 544:549 arg1 spectrometry 556:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 6 22 theme structure 1109:1117 arg1 sites 1076:1080 glycan attachment sites 1058:1080 glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1058:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 22 theme structure 1109:1117 arg1 structure 1109:1117 the NeuAcHexHexNAcHex-O structure 1085:1117 the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1085:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 23 theme skeletal 1279:1286 arg1 muscle 1288:1293 human skeletal muscle 1273:1293 human skeletal muscle 1273:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 3 24 gly glycopeptides 595:607 arg2 glycopeptides 595:607 glycopeptides 595:607 glycopeptides 595:607 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 5 25 theme occupancy 1013:1021 arg1 level 985:989 level 985:989 level of glycosylation site occupancy 985:1021 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 5 25 theme occupancy 1013:1021 arg1 structures 970:979 glycan core structures 958:979 glycan core structures 958:979 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 7 26 theme complex 1401:1407 arg1 O-glycosylation 1417:1431 the complex in vivo O-glycosylation 1397:1431 the complex in vivo O-glycosylation of alpha-dystroglycan 1397:1453 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 2 27 gly glycopeptides 453:465 arg1 alpha-dystroglycan 470:487 alpha-dystroglycan 470:487 alpha-dystroglycan 470:487 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 2 27 gly glycopeptides 453:465 arg2 glycopeptides 453:465 tryptic glycopeptides 445:465 tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle 445:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 2 27 gly glycopeptides 453:465 arg1 muscle 509:514 human skeletal muscle 494:514 human skeletal muscle 494:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 5 28 theme glycosylation 994:1006 arg1 occupancy 1013:1021 glycosylation site occupancy 994:1021 glycosylation site occupancy 994:1021 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 4 29 theme sialic 856:861 arg1 acid 863:866 sialic acid 856:866 sialic acid 856:866 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 1 30 theme ligand-binding 220:233 arg1 activity 235:242 its ligand-binding activity 216:242 its ligand-binding activity 216:242 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 0 31 theme O-glycan 34:41 arg1 structures 43:52 site-specific O-glycan structures 20:52 site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle 20:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 1 32 theme congenital 318:327 arg1 dystrophies 338:348 congenital muscular dystrophies 318:348 congenital muscular dystrophies 318:348 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 1 33 gly glycosylation 137:149 arg1 alpha-dystroglycan 180:197 the extracellular protein alpha-dystroglycan 154:197 the extracellular protein alpha-dystroglycan 154:197 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 3 34 used used 573:576 arg2 spectrometry 556:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 6 35 theme human 1273:1277 arg1 muscle 1288:1293 human skeletal muscle 1273:1293 human skeletal muscle 1273:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 36 theme human 1243:1247 arg1 alpha-dystroglycan 1249:1266 human alpha-dystroglycan 1243:1266 human alpha-dystroglycan from human skeletal muscle 1243:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 0 37 theme site-specific 20:32 arg1 structures 43:52 site-specific O-glycan structures 20:52 site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle 20:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 1 38 theme muscular 329:336 arg1 dystrophies 338:348 congenital muscular dystrophies 318:348 congenital muscular dystrophies 318:348 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 4 39 dep had 719:721 arg1 elongated 798:806 elongated 798:806 elongated 798:806 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 5 40 theme glycan 958:963 arg1 structures 970:979 glycan core structures 958:979 glycan core structures 958:979 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 2 41 theme skeletal 500:507 arg1 muscle 509:514 human skeletal muscle 494:514 human skeletal muscle 494:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 5 42 theme Hex-O-Thr 899:907 arg1 glycopeptides 909:921 Hex-O-Thr glycopeptides 899:921 Hex-O-Thr glycopeptides 899:921 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 1 43 theme dystrophies 338:348 arg1 forms 309:313 several forms 301:313 several forms of congenital muscular dystrophies 301:348 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 3 44 theme mass 551:554 arg1 spectrometry 556:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 2 45 theme tryptic 445:451 arg1 glycopeptides 453:465 tryptic glycopeptides 445:465 tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle 445:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 2 46 theme human 494:498 arg1 muscle 509:514 human skeletal muscle 494:514 human skeletal muscle 494:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 0 47 theme structures 43:52 arg1 Characterization 0:15 Characterization 0:15 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 0:131 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 6 48 theme NeuAcHexHexNAcHex-O 1089:1107 arg1 structure 1109:1117 the NeuAcHexHexNAcHex-O structure 1085:1117 the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1085:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 4 49 contain had 719:721 arg2 HexNAc-O-Ser/Thr 743:758 HexNAc-O-Ser/Thr 743:758 HexNAc-O-Ser/Thr 743:758 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 4 49 contain had 719:721 arg2 Hex-O-Thr 730:738 Hex-O-Thr 730:738 Hex-O-Thr 730:738 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 4 49 contain had 719:721 arg1 O-glycans 703:711 The O-glycans 699:711 The O-glycans found 699:717 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 1 50 theme altered 249:255 arg1 glycosylation 268:280 altered or blocked glycosylation 249:280 altered or blocked glycosylation 249:280 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 0 51 theme mucin-like 65:74 arg1 alpha-dystroglycan 86:103 alpha-dystroglycan 86:103 alpha-dystroglycan 86:103 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 0 51 theme mucin-like 65:74 arg1 domain 76:81 the mucin-like domain 61:81 the mucin-like domain of alpha-dystroglycan from human skeletal muscle 61:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 7 52 dep in 1409:1410 arg1 vivo 1412:1415 vivo 1412:1415 vivo 1412:1415 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 3 53 theme chromatography 529:542 arg1 spectrometry 556:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry 517:567 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 2 54 theme enrichment 410:419 arg1 strategies 421:430 capture-and-release enrichment strategies 390:430 capture-and-release enrichment strategies 390:430 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 6 55 theme alpha-dystroglycan 1249:1266 arg1 domain 1233:1238 the mucin-like domain 1218:1238 the mucin-like domain of human alpha-dystroglycan from human skeletal muscle 1218:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 55 theme alpha-dystroglycan 1249:1266 arg1 alpha-dystroglycan 1249:1266 human alpha-dystroglycan 1243:1266 human alpha-dystroglycan from human skeletal muscle 1243:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 7 56 theme human 1346:1350 arg1 alpha-dystroglycan 1352:1369 human alpha-dystroglycan 1346:1369 human alpha-dystroglycan 1346:1369 Twenty-five glycopeptides were characterized from human alpha-dystroglycan, which provide insight to the complex in vivo O-glycosylation of alpha-dystroglycan. 20507882 2 57 theme capture-and-release 390:408 arg1 strategies 421:430 capture-and-release enrichment strategies 390:430 capture-and-release enrichment strategies 390:430 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 3 58 theme alpha-dystroglycan 679:696 arg1 alpha-dystroglycan 679:696 alpha-dystroglycan 679:696 alpha-dystroglycan 679:696 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 3 58 theme alpha-dystroglycan 679:696 arg1 domain 669:674 the mucin-like and C-terminal domain 639:674 domain 669:674 Nano-liquid chromatography tandem mass spectrometry was used to identify both glycopeptides and peptides corresponding to the mucin-like and C-terminal domain of alpha-dystroglycan. 20507882 0 59 from muscle 125:130 arg1 alpha-dystroglycan 86:103 alpha-dystroglycan 86:103 alpha-dystroglycan 86:103 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 0 59 from muscle 125:130 arg1 domain 76:81 the mucin-like domain 61:81 the mucin-like domain of alpha-dystroglycan from human skeletal muscle 61:130 Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. 20507882 5 60 gly glycopeptides 909:921 arg2 glycopeptides 909:921 Hex-O-Thr glycopeptides 899:921 Hex-O-Thr glycopeptides 899:921 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 4 61 dep Hex-O-Thr 730:738 arg1 structures 769:778 anchored structures 760:778 anchored structures 760:778 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 4 62 dep elongated 798:806 arg1 terminated 840:849 terminated 840:849 terminated with sialic acid 840:866 The O-glycans found had either Hex-O-Thr or HexNAc-O-Ser/Thr anchored structures, which were often elongated and frequently, but not always, terminated with sialic acid. 20507882 2 63 theme sialic 378:383 arg1 acid 385:388 sialic acid 378:388 sialic acid 378:388 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 1 64 theme blocked 260:266 arg1 glycosylation 268:280 altered or blocked glycosylation 249:280 altered or blocked glycosylation 249:280 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 6 65 theme mucin-like 1222:1231 arg1 domain 1233:1238 the mucin-like domain 1218:1238 the mucin-like domain of human alpha-dystroglycan from human skeletal muscle 1218:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 65 theme mucin-like 1222:1231 arg1 alpha-dystroglycan 1249:1266 human alpha-dystroglycan 1243:1266 human alpha-dystroglycan from human skeletal muscle 1243:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 66 from muscle 1288:1293 arg1 domain 1233:1238 the mucin-like domain 1218:1238 the mucin-like domain of human alpha-dystroglycan from human skeletal muscle 1218:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 66 from muscle 1288:1293 arg1 alpha-dystroglycan 1249:1266 human alpha-dystroglycan 1243:1266 human alpha-dystroglycan from human skeletal muscle 1243:1293 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 1 67 theme extracellular 158:170 arg1 alpha-dystroglycan 180:197 the extracellular protein alpha-dystroglycan 154:197 the extracellular protein alpha-dystroglycan 154:197 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 20507882 5 68 theme site 1008:1011 arg1 occupancy 1013:1021 glycosylation site occupancy 994:1021 glycosylation site occupancy 994:1021 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 6 69 theme attachment 1065:1074 arg1 sites 1076:1080 glycan attachment sites 1058:1080 glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1058:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 6 69 theme attachment 1065:1074 arg1 structure 1109:1117 the NeuAcHexHexNAcHex-O structure 1085:1117 the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man) 1085:1208 We demonstrate for the first time glycan attachment sites of the NeuAcHexHexNAcHex-O structure corresponding to the anticipated Neu5Acalpha3Galbeta4GlcNAcbeta2Man-O-glycan (sLacNAc-Man), within the mucin-like domain of human alpha-dystroglycan from human skeletal muscle. 20507882 5 70 theme core 965:968 arg1 structures 970:979 glycan core structures 958:979 glycan core structures 958:979 The HexNAc-O-Ser/Thr, but not Hex-O-Thr glycopeptides, displayed heterogeneity regarding glycan core structures and level of glycosylation site occupancy. 20507882 2 71 from muscle 509:514 arg1 glycopeptides 453:465 tryptic glycopeptides 445:465 tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle 445:514 By immunoprecipitation and sialic acid capture-and-release enrichment strategies, we isolated tryptic glycopeptides of alpha-dystroglycan from human skeletal muscle. 20507882 1 72 theme protein 172:178 arg1 alpha-dystroglycan 180:197 the extracellular protein alpha-dystroglycan 154:197 the extracellular protein alpha-dystroglycan 154:197 The glycosylation of the extracellular protein alpha-dystroglycan is important for its ligand-binding activity, and altered or blocked glycosylation is associated with several forms of congenital muscular dystrophies. 21763278 0 0 from processing 18:27 arg1 cardiomyocytes 66:79 cardiomyocytes 66:79 cardiomyocytes 66:79 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 13 1 from Lys-79 1354:1359 arg1 mutations 1322:1330 single or double mutations 1305:1330 single or double mutations at Arg-73, Arg-76 and Lys-79 1305:1359 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 7 2 theme pro-BNP 740:746 arg1 processing 748:757 pro-BNP processing 740:757 pro-BNP processing in cells 740:766 The effects of furin and corin expression on pro-BNP processing in cells also were examined. 21763278 7 3 from effects 699:705 arg1 processing 748:757 pro-BNP processing 740:757 pro-BNP processing in cells 740:766 The effects of furin and corin expression on pro-BNP processing in cells also were examined. 21763278 4 4 theme murine 503:508 arg1 cardiomyocytes 515:528 murine HL-1 cardiomyocytes 503:528 murine HL-1 cardiomyocytes 503:528 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 9 5 from Thr-71 928:933 arg1 Mutation 916:923 Mutation 916:923 Mutation at Thr-71 916:933 Mutation at Thr-71 reduced O-glycans on pro-BNP and increased pro-BNP processing. 21763278 7 6 theme furin 710:714 arg1 effects 699:705 The effects 695:705 The effects of furin and corin expression on pro-BNP processing in cells 695:766 The effects of furin and corin expression on pro-BNP processing in cells also were examined. 21763278 15 7 gly glycosylation 1520:1532 arg1 cardiomyocytes 1537:1550 cardiomyocytes 1537:1550 cardiomyocytes 1537:1550 Our data indicate that pro-BNP glycosylation in cardiomyocytes differed significantly from that in HEK 293 cells. 21763278 13 8 theme single 1305:1310 arg1 mutations 1322:1330 single or double mutations 1305:1330 single or double mutations at Arg-73, Arg-76 and Lys-79 1305:1359 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 0 9 from Glycosylation 0:12 arg1 cardiomyocytes 66:79 cardiomyocytes 66:79 cardiomyocytes 66:79 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 13 10 from Arg-76 1343:1348 arg1 mutations 1322:1330 single or double mutations 1305:1330 single or double mutations at Arg-73, Arg-76 and Lys-79 1305:1359 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 6 11 theme glycosylation 623:635 arg1 inhibitors 637:646 glycosylation inhibitors 623:646 glycosylation inhibitors 623:646 Glycosidases and glycosylation inhibitors were used to examine carbohydrates on pro-BNP. 21763278 15 12 theme HEK 1588:1590 arg1 cells 1596:1600 HEK 293 cells 1588:1600 HEK 293 cells 1588:1600 Our data indicate that pro-BNP glycosylation in cardiomyocytes differed significantly from that in HEK 293 cells. 21763278 15 13 from glycosylation 1520:1532 arg1 cardiomyocytes 1537:1550 cardiomyocytes 1537:1550 cardiomyocytes 1537:1550 Our data indicate that pro-BNP glycosylation in cardiomyocytes differed significantly from that in HEK 293 cells. 21763278 12 14 from Arg-73 1185:1190 arg1 Mutations 1172:1180 Mutations 1172:1180 Mutations at Arg-73 and Arg-76 1172:1201 Mutations at Arg-73 and Arg-76, but not Lys-79, prevented pro-BNP processing. 21763278 8 15 theme HEK 805:807 arg1 cells 813:817 HEK 293 cells 805:817 HEK 293 cells 805:817 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 9 16 theme increased 968:976 arg1 processing 986:995 increased pro-BNP processing 968:995 increased pro-BNP processing 968:995 Mutation at Thr-71 reduced O-glycans on pro-BNP and increased pro-BNP processing. 21763278 11 17 theme HEK 1125:1127 arg1 cells 1133:1137 HEK 293 cells 1125:1137 HEK 293 cells 1125:1137 In HEK 293 cells, pro-BNP was processed by furin. 21763278 1 18 theme B-type 82:87 arg1 biomarkers 144:153 biomarkers 144:153 biomarkers for the diagnosis of heart failure 144:188 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 18 theme B-type 82:87 arg1 peptides 131:138 its related peptides 119:138 its related peptides 119:138 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 18 theme B-type 82:87 arg1 BNP 110:112 BNP 110:112 BNP 110:112 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 18 theme B-type 82:87 arg1 peptide 101:107 B-type natriuretic peptide 82:107 B-type natriuretic peptide (BNP) 82:113 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 8 19 theme O-glycans 873:881 arg1 O-glycans 873:881 O-glycans 873:881 O-glycans 873:881 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 8 19 theme O-glycans 873:881 arg1 amounts 862:868 significant amounts 850:868 significant amounts of O-glycans with terminal oligosialic acids 850:913 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 6 20 from carbohydrates 669:681 arg1 pro-BNP 686:692 pro-BNP 686:692 pro-BNP 686:692 Glycosidases and glycosylation inhibitors were used to examine carbohydrates on pro-BNP. 21763278 0 21 theme peptide 55:61 arg1 processing 18:27 processing 18:27 processing 18:27 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 0 21 theme peptide 55:61 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation 0:12 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 3 22 from processing 385:394 arg1 cardiomyocytes 410:423 cardiomyocytes 410:423 cardiomyocytes 410:423 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 1 23 theme natriuretic 89:99 arg1 biomarkers 144:153 biomarkers 144:153 biomarkers for the diagnosis of heart failure 144:188 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 23 theme natriuretic 89:99 arg1 peptides 131:138 its related peptides 119:138 its related peptides 119:138 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 23 theme natriuretic 89:99 arg1 BNP 110:112 BNP 110:112 BNP 110:112 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 23 theme natriuretic 89:99 arg1 peptide 101:107 B-type natriuretic peptide 82:107 B-type natriuretic peptide (BNP) 82:113 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 8 24 theme oligosialic 897:907 arg1 acids 909:913 terminal oligosialic acids 888:913 terminal oligosialic acids 888:913 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 2 25 from sites 241:245 arg1 pro-BNP 275:281 human pro-BNP 269:281 human pro-BNP 269:281 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 1 26 theme heart 176:180 arg1 failure 182:188 heart failure 176:188 heart failure 176:188 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 10 27 theme pro-BNP 1069:1075 arg1 WT 1077:1078 pro-BNP WT 1069:1078 pro-BNP WT 1069:1078 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 10 28 theme T71A 1084:1087 arg1 mutant 1089:1094 T71A mutant 1084:1094 T71A mutant 1084:1094 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 13 29 theme HL-1 1253:1256 arg1 cardiomyocytes 1258:1271 HL-1 cardiomyocytes 1253:1271 HL-1 cardiomyocytes 1253:1271 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 1 30 theme failure 182:188 arg1 diagnosis 163:171 the diagnosis 159:171 the diagnosis of heart failure 159:188 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 3 31 gly glycosylation 355:367 arg1 study 336:340 this study 331:340 this study 331:340 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 3 31 gly glycosylation 355:367 arg1 cardiomyocytes 410:423 cardiomyocytes 410:423 cardiomyocytes 410:423 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 3 31 gly glycosylation 355:367 arg1 pro-BNP 399:405 pro-BNP 399:405 pro-BNP 399:405 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 5 32 theme Western 588:594 arg1 blotting 596:603 Western blotting 588:603 Western blotting 588:603 Pro-BNP and BNP were analyzed by immunoprecipitation and Western blotting. 21763278 13 33 theme double 1315:1320 arg1 mutations 1322:1330 single or double mutations 1305:1330 single or double mutations at Arg-73, Arg-76 and Lys-79 1305:1359 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 10 34 theme HL-1 1001:1004 arg1 cardiomyocytes 1006:1019 HL-1 cardiomyocytes 1001:1019 HL-1 cardiomyocytes 1001:1019 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 3 35 theme proteolytic 373:383 arg1 processing 385:394 proteolytic processing 373:394 proteolytic processing 373:394 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 13 36 theme pro-BNP 1377:1383 arg1 processing 1385:1394 pro-BNP processing 1377:1394 pro-BNP processing 1377:1394 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 2 37 theme functional 291:300 arg1 significance 302:313 the functional significance 287:313 the functional significance 287:313 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 2 37 theme functional 291:300 arg1 unclear 319:325 unclear 319:325 unclear 319:325 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 8 38 theme terminal 888:895 arg1 acids 909:913 terminal oligosialic acids 888:913 terminal oligosialic acids 888:913 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 0 39 theme natriuretic 43:53 arg1 peptide 55:61 pro-B-type natriuretic peptide 32:61 pro-B-type natriuretic peptide 32:61 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 12 40 theme pro-BNP 1230:1236 arg1 processing 1238:1247 pro-BNP processing 1230:1247 pro-BNP processing 1230:1247 Mutations at Arg-73 and Arg-76, but not Lys-79, prevented pro-BNP processing. 21763278 15 41 theme pro-BNP 1512:1518 arg1 glycosylation 1520:1532 pro-BNP glycosylation 1512:1532 pro-BNP glycosylation in cardiomyocytes 1512:1550 Our data indicate that pro-BNP glycosylation in cardiomyocytes differed significantly from that in HEK 293 cells. 21763278 7 42 theme corin 720:724 arg1 effects 699:705 The effects 695:705 The effects of furin and corin expression on pro-BNP processing in cells 695:766 The effects of furin and corin expression on pro-BNP processing in cells also were examined. 21763278 0 43 gly Glycosylation 0:12 arg1 peptide 55:61 pro-B-type natriuretic peptide 32:61 pro-B-type natriuretic peptide 32:61 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 0 44 theme pro-B-type 32:41 arg1 peptide 55:61 pro-B-type natriuretic peptide 32:61 pro-B-type natriuretic peptide 32:61 Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes. 21763278 6 45 used used 653:656 arg2 Glycosidases 606:617 Glycosidases 606:617 Glycosidases 606:617 Glycosidases and glycosylation inhibitors were used to examine carbohydrates on pro-BNP. 21763278 6 45 used used 653:656 arg2 inhibitors 637:646 glycosylation inhibitors 623:646 glycosylation inhibitors 623:646 Glycosidases and glycosylation inhibitors were used to examine carbohydrates on pro-BNP. 21763278 7 46 from processing 748:757 arg1 cells 762:766 cells 762:766 cells 762:766 The effects of furin and corin expression on pro-BNP processing in cells also were examined. 21763278 8 47 contain contained 840:848 arg2 pro-BNP 832:838 recombinant pro-BNP 820:838 recombinant pro-BNP 820:838 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 8 47 contain contained 840:848 arg1 cells 813:817 HEK 293 cells 805:817 HEK 293 cells 805:817 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 8 47 contain contained 840:848 arg1 pro-BNP 832:838 recombinant pro-BNP 820:838 recombinant pro-BNP 820:838 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 8 47 contain contained 840:848 arg2 amounts 862:868 significant amounts 850:868 significant amounts of O-glycans with terminal oligosialic acids 850:913 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 8 47 contain contained 840:848 arg2 O-glycans 873:881 O-glycans 873:881 O-glycans 873:881 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 9 48 theme pro-BNP 978:984 arg1 processing 986:995 increased pro-BNP processing 968:995 increased pro-BNP processing 968:995 Mutation at Thr-71 reduced O-glycans on pro-BNP and increased pro-BNP processing. 21763278 3 49 theme pro-BNP 399:405 arg1 processing 385:394 proteolytic processing 373:394 proteolytic processing 373:394 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 3 49 theme pro-BNP 399:405 arg1 glycosylation 355:367 glycosylation 355:367 glycosylation 355:367 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 4 50 theme HL-1 510:513 arg1 cardiomyocytes 515:528 murine HL-1 cardiomyocytes 503:528 murine HL-1 cardiomyocytes 503:528 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 13 51 from Arg-73 1335:1340 arg1 mutations 1322:1330 single or double mutations 1305:1330 single or double mutations at Arg-73, Arg-76 and Lys-79 1305:1359 In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. 21763278 3 52 from glycosylation 355:367 arg1 cardiomyocytes 410:423 cardiomyocytes 410:423 cardiomyocytes 410:423 In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. 21763278 2 53 theme O-glycosylation 225:239 arg1 sites 241:245 several O-glycosylation sites 217:245 several O-glycosylation sites 217:245 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 2 53 theme O-glycosylation 225:239 arg1 Thr-71 258:263 Thr-71 258:263 Thr-71 258:263 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 4 54 theme HEK 485:487 arg1 cells 493:497 HEK 293 cells 485:497 HEK 293 cells 485:497 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 2 55 gly O-glycosylation 225:239 arg2 Thr-71 258:263 Thr-71 258:263 Thr-71 258:263 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 2 55 gly O-glycosylation 225:239 arg2 sites 241:245 several O-glycosylation sites 217:245 several O-glycosylation sites 217:245 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 16 56 theme HEK 1606:1608 arg1 cells 1614:1618 HEK 293 cells 1606:1618 HEK 293 cells 1606:1618 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 21763278 8 57 with amounts 862:868 arg1 acids 909:913 terminal oligosialic acids 888:913 terminal oligosialic acids 888:913 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 4 58 theme pro-BNP 432:438 arg1 wild-type 440:448 Human pro-BNP wild-type 426:448 Human pro-BNP wild-type (WT) 426:453 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 4 58 theme pro-BNP 432:438 arg1 WT 451:452 WT 451:452 WT 451:452 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 2 59 theme several 217:223 arg1 sites 241:245 several O-glycosylation sites 217:245 several O-glycosylation sites 217:245 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 2 59 theme several 217:223 arg1 Thr-71 258:263 Thr-71 258:263 Thr-71 258:263 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 16 60 dep corin 1679:1683 arg1 cleaved 1685:1691 cleaved 1685:1691 corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79 1679:1756 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 21763278 10 61 theme little 1047:1052 arg1 O-glycans 1054:1062 little O-glycans 1047:1062 little O-glycans 1047:1062 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 10 62 contain contained 1037:1045 arg2 Thr-71 1030:1035 residue Thr-71 1022:1035 residue Thr-71 1022:1035 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 10 62 contain contained 1037:1045 arg2 O-glycans 1054:1062 little O-glycans 1047:1062 little O-glycans 1047:1062 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 10 62 contain contained 1037:1045 arg1 cardiomyocytes 1006:1019 HL-1 cardiomyocytes 1001:1019 HL-1 cardiomyocytes 1001:1019 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 10 62 contain contained 1037:1045 arg1 Thr-71 1030:1035 residue Thr-71 1022:1035 residue Thr-71 1022:1035 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 4 63 theme Human 426:430 arg1 wild-type 440:448 Human pro-BNP wild-type 426:448 Human pro-BNP wild-type (WT) 426:453 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 4 63 theme Human 426:430 arg1 WT 451:452 WT 451:452 WT 451:452 Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. 21763278 1 64 theme related 123:129 arg1 biomarkers 144:153 biomarkers 144:153 biomarkers for the diagnosis of heart failure 144:188 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 64 theme related 123:129 arg1 peptides 131:138 its related peptides 119:138 its related peptides 119:138 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 1 64 theme related 123:129 arg1 peptide 101:107 B-type natriuretic peptide 82:107 B-type natriuretic peptide (BNP) 82:113 B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. 21763278 2 65 theme human 269:273 arg1 pro-BNP 275:281 human pro-BNP 269:281 human pro-BNP 269:281 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 2 66 theme Recent 191:196 arg1 studies 198:204 Recent studies 191:204 Recent studies 191:204 Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. 21763278 10 67 theme residue 1022:1028 arg1 Thr-71 1030:1035 residue Thr-71 1022:1035 residue Thr-71 1022:1035 In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. 21763278 7 68 dep furin 710:714 arg1 expression 726:735 expression 726:735 expression 726:735 The effects of furin and corin expression on pro-BNP processing in cells also were examined. 21763278 12 69 from Arg-76 1196:1201 arg1 Mutations 1172:1180 Mutations 1172:1180 Mutations at Arg-73 and Arg-76 1172:1201 Mutations at Arg-73 and Arg-76, but not Lys-79, prevented pro-BNP processing. 21763278 14 70 theme pro-BNP 1450:1456 arg1 processing 1458:1467 pro-BNP processing 1450:1467 pro-BNP processing 1450:1467 Only when all these three residues were mutated, was pro-BNP processing completely blocked. 21763278 8 71 theme recombinant 820:830 arg1 pro-BNP 832:838 recombinant pro-BNP 820:838 recombinant pro-BNP 820:838 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 16 72 from Arg-76 1646:1651 arg1 pro-BNP 1635:1641 pro-BNP 1635:1641 pro-BNP at Arg-76 1635:1651 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 21763278 8 73 theme significant 850:860 arg1 O-glycans 873:881 O-glycans 873:881 O-glycans 873:881 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 8 73 theme significant 850:860 arg1 amounts 862:868 significant amounts 850:868 significant amounts of O-glycans with terminal oligosialic acids 850:913 We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. 21763278 16 74 theme multiple 1704:1711 arg1 Arg-73 1732:1737 Arg-73 1732:1737 Arg-73 1732:1737 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 21763278 16 74 theme multiple 1704:1711 arg1 residues 1713:1720 multiple residues 1704:1720 multiple residues including Arg-73, Arg-76 and Lys-79 1704:1756 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 21763278 16 74 theme multiple 1704:1711 arg1 Lys-79 1751:1756 Lys-79 1751:1756 Lys-79 1751:1756 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 21763278 16 74 theme multiple 1704:1711 arg1 Arg-76 1740:1745 Arg-76 1740:1745 Arg-76 1740:1745 In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79. 15379548 10 0 theme ZP3 1843:1845 arg1 proteins 1847:1854 human and mouse ZP3 proteins 1827:1854 human and mouse ZP3 proteins 1827:1854 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 7 1 theme C-terminal 1159:1168 arg1 peptides 1170:1177 several C-terminal peptides 1151:1177 several C-terminal peptides 1151:1177 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 4 2 theme proteins 532:539 arg1 structure 489:497 secondary structure 479:497 secondary structure 479:497 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 4 2 theme proteins 532:539 arg1 processing 503:512 processing 503:512 processing 503:512 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 8 3 from residues 1274:1281 arg1 domain 1305:1310 the conserved zona domain 1286:1310 the conserved zona domain 1286:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 6 4 theme mass 870:873 arg1 spectrometry 875:886 LC-QTOF mass spectrometry 862:886 LC-QTOF mass spectrometry 862:886 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 2 5 gly glycoproteins 285:297 arg1 glycoproteins 285:297 three glycoproteins 279:297 three glycoproteins (ZP1, ZP2, ZP3) 279:313 The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2, ZP3). 15379548 9 6 theme O-glycans 1709:1717 arg1 clusters 1697:1704 clusters 1697:1704 clusters of O-glycans 1697:1717 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 5 7 theme taxon-specific 617:630 arg1 recognition 642:652 taxon-specific sperm-egg recognition 617:652 taxon-specific sperm-egg recognition 617:652 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 8 8 theme additional 1435:1444 arg1 bonds 1456:1460 two additional disulfide bonds 1431:1460 two additional disulfide bonds 1431:1460 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 9 theme bonds 1456:1460 arg1 indeterminate 1466:1478 indeterminate 1466:1478 indeterminate 1466:1478 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 9 theme bonds 1456:1460 arg1 linkage 1420:1426 the precise linkage 1408:1426 the precise linkage of two additional disulfide bonds 1408:1460 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 6 10 theme complete 843:850 arg1 coverage 852:859 nearly complete coverage 836:859 nearly complete coverage 836:859 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 11 theme N-terminal 929:938 arg1 peptide 947:953 an N-terminal signal peptide 926:953 an N-terminal signal peptide (amino acids 1-22) 926:972 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 11 theme N-terminal 929:938 arg1 acids 962:966 amino acids 1-22 956:971 amino acids 1-22 956:971 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 4 12 theme biological 578:587 arg1 material 589:596 biological material 578:596 biological material 578:596 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 5 13 theme structural 672:681 arg1 differences 683:693 structural differences 672:693 structural differences in a zona protein required for matrix formation 672:741 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 5 14 theme mouse 820:824 arg1 ZP3 826:828 mouse ZP3 820:828 mouse ZP3 820:828 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 10 15 theme binding 1928:1934 arg1 explanations 1891:1902 alternative explanations 1879:1902 alternative explanations of taxon-specific sperm binding 1879:1934 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 8 16 theme Cys 1365:1367 arg1 /Cys 1337:1340 Cys(46)/Cys(140) 1330:1345 Cys(46)/Cys(140) 1330:1345 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 16 theme Cys 1365:1367 arg1 /Cys 1373:1376 Cys(217)/Cys(282) 1365:1381 Cys(217)/Cys(282) 1365:1381 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 9 17 theme potential 1598:1606 arg1 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 8 18 theme remaining 1505:1513 arg1 residues 1529:1536 the remaining four cysteine residues 1501:1536 the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)) 1501:1577 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 10 19 theme taxon-specific 1907:1920 arg1 binding 1928:1934 taxon-specific sperm binding 1907:1934 taxon-specific sperm binding 1907:1934 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 9 20 theme oligosaccharide 1617:1631 arg1 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 8 21 theme cysteine 1520:1527 arg1 residues 1529:1536 the remaining four cysteine residues 1501:1536 the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)) 1501:1577 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 22 dep ascertained 1317:1327 arg1 /Cys 1337:1340 Cys(46)/Cys(140) 1330:1345 Cys(46)/Cys(140) 1330:1345 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 22 dep ascertained 1317:1327 arg1 /Cys 1392:1395 Cys(239)/Cys(300) 1384:1400 Cys(239)/Cys(300) 1384:1400 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 22 dep ascertained 1317:1327 arg1 /Cys 1355:1358 Cys(78)/Cys(99) 1348:1362 Cys(78)/Cys(99) 1348:1362 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 22 dep ascertained 1317:1327 arg1 /Cys 1373:1376 Cys(217)/Cys(282) 1365:1381 Cys(217)/Cys(282) 1365:1381 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 9 23 theme sites 1641:1645 arg1 Three 1580:1584 Three 1580:1584 Three 1580:1584 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 23 theme sites 1641:1645 arg1 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 7 24 theme several 1151:1157 arg1 peptides 1170:1177 several C-terminal peptides 1151:1177 several C-terminal peptides 1151:1177 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 2 25 theme mouse 243:247 arg1 pellucida 254:262 The mouse zona pellucida 239:262 The mouse zona pellucida 239:262 The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2, ZP3). 15379548 2 26 dep glycoproteins 285:297 arg1 ZP1 300:302 ZP1 300:302 ZP1 300:302 The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2, ZP3). 15379548 2 26 dep glycoproteins 285:297 arg1 ZP3 310:312 ZP3 310:312 ZP3 310:312 The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2, ZP3). 15379548 2 26 dep glycoproteins 285:297 arg1 ZP2 305:307 ZP2 305:307 ZP2 305:307 The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2, ZP3). 15379548 0 27 theme CHO 89:91 arg1 cells 93:97 glycosylation-deficient CHO cells 65:97 glycosylation-deficient CHO cells 65:97 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 6 28 theme amino 956:960 arg1 peptide 947:953 an N-terminal signal peptide 926:953 an N-terminal signal peptide (amino acids 1-22) 926:972 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 28 theme amino 956:960 arg1 acids 962:966 amino acids 1-22 956:971 amino acids 1-22 956:971 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 7 29 dep pyroglutamate 1119:1131 arg1 glutamine 1093:1101 The resultant N-terminal glutamine 1068:1101 The resultant N-terminal glutamine 1068:1101 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 29 dep pyroglutamate 1119:1131 arg1 to 1116:1117 to 1116:1117 to 1116:1117 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 29 dep pyroglutamate 1119:1131 arg1 pyrGln 1134:1139 pyrGln(23) 1134:1143 pyrGln(23) 1134:1143 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 29 dep pyroglutamate 1119:1131 arg1 pyroglutamate 1119:1131 pyroglutamate 1119:1131 pyroglutamate 1119:1131 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 5 30 theme zona 700:703 arg1 protein 705:711 a zona protein 698:711 a zona protein required for matrix formation 698:741 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 6 31 theme peptide 947:953 arg1 release 982:988 the release 978:988 the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424) 978:1065 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 31 theme peptide 947:953 arg1 cleavage 914:921 the cleavage 910:921 the cleavage of an N-terminal signal peptide (amino acids 1-22) 910:972 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 32 used used 892:895 arg2 spectrometry 875:886 LC-QTOF mass spectrometry 862:886 LC-QTOF mass spectrometry 862:886 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 0 33 theme human 42:46 arg1 ZP3 48:50 recombinant human ZP3 30:50 recombinant human ZP3 30:50 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 6 34 theme C-terminal 1013:1022 arg1 domain 1038:1043 a C-terminal transmembrane domain 1011:1043 a C-terminal transmembrane domain (amino acids 379-424) 1011:1065 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 34 theme C-terminal 1013:1022 arg1 acids 1052:1056 amino acids 379-424 1046:1064 amino acids 379-424 1046:1064 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 8 35 theme residues 1274:1281 arg1 linkages 1247:1254 The disulfide bond linkages 1228:1254 The disulfide bond linkages of eight cysteine residues in the conserved zona domain 1228:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 3 36 theme acid 385:388 arg1 sequence 390:397 the cDNA nucleic acid sequence 368:397 the cDNA nucleic acid sequence 368:397 The primary structure of each has been deduced from the cDNA nucleic acid sequence, and each has been analyzed by mass spectrometry. 15379548 0 37 theme Mass 0:3 arg1 analysis 18:25 Mass spectrometry analysis 0:25 Mass spectrometry analysis of recombinant human ZP3 0:50 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 3 38 theme each 341:344 arg1 structure 328:336 The primary structure 316:336 The primary structure of each 316:344 The primary structure of each has been deduced from the cDNA nucleic acid sequence, and each has been analyzed by mass spectrometry. 15379548 4 39 theme processing 503:512 arg1 determination 458:470 determination 458:470 determination of the secondary structure and processing of the human zona proteins 458:539 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 6 40 theme secreted 993:1000 arg1 ZP3 1002:1004 secreted ZP3 993:1004 secreted ZP3 993:1004 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 3 41 theme cDNA 372:375 arg1 sequence 390:397 the cDNA nucleic acid sequence 368:397 the cDNA nucleic acid sequence 368:397 The primary structure of each has been deduced from the cDNA nucleic acid sequence, and each has been analyzed by mass spectrometry. 15379548 8 42 theme zona 1300:1303 arg1 domain 1305:1310 the conserved zona domain 1286:1310 the conserved zona domain 1286:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 9 43 located observed 1724:1731 arg1 regions 1744:1750 two regions 1740:1750 two regions 1740:1750 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 43 located observed 1724:1731 arg1 acids 1759:1763 amino acids 156-173 and 260-281 1753:1783 amino acids 156-173 and 260-281 1753:1783 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 43 located observed 1724:1731 arg2 clusters 1697:1704 clusters 1697:1704 clusters of O-glycans 1697:1717 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 5 44 theme human 756:760 arg1 ZP3 762:764 recombinant human ZP3 744:764 recombinant human ZP3 744:764 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 0 45 theme recombinant 30:40 arg1 ZP3 48:50 recombinant human ZP3 30:50 recombinant human ZP3 30:50 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 5 46 theme matrix 726:731 arg1 formation 733:741 matrix formation 726:741 matrix formation 726:741 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 0 47 theme ZP3 48:50 arg1 analysis 18:25 Mass spectrometry analysis 0:25 Mass spectrometry analysis of recombinant human ZP3 0:50 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 4 48 theme structure 489:497 arg1 determination 458:470 determination 458:470 determination of the secondary structure and processing of the human zona proteins 458:539 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 1 49 theme mouse 227:231 arg1 eggs 233:236 mouse eggs 227:236 mouse eggs 227:236 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 6 50 from domain 1038:1043 arg1 release 982:988 the release 978:988 the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424) 978:1065 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 50 from domain 1038:1043 arg1 cleavage 914:921 the cleavage 910:921 the cleavage of an N-terminal signal peptide (amino acids 1-22) 910:972 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 51 theme amino 1046:1050 arg1 domain 1038:1043 a C-terminal transmembrane domain 1011:1043 a C-terminal transmembrane domain (amino acids 379-424) 1011:1065 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 51 theme amino 1046:1050 arg1 acids 1052:1056 amino acids 379-424 1046:1064 amino acids 379-424 1046:1064 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 5 52 from differences 683:693 arg1 protein 705:711 a zona protein 698:711 a zona protein required for matrix formation 698:741 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 10 53 theme mouse 1837:1841 arg1 proteins 1847:1854 human and mouse ZP3 proteins 1827:1854 human and mouse ZP3 proteins 1827:1854 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 4 54 theme zona 527:530 arg1 proteins 532:539 the human zona proteins 517:539 the human zona proteins 517:539 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 6 55 theme LC-QTOF 862:868 arg1 spectrometry 875:886 LC-QTOF mass spectrometry 862:886 LC-QTOF mass spectrometry 862:886 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 10 56 theme human 1827:1831 arg1 proteins 1847:1854 human and mouse ZP3 proteins 1827:1854 human and mouse ZP3 proteins 1827:1854 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 8 57 theme disulfide 1232:1240 arg1 linkages 1247:1254 The disulfide bond linkages 1228:1254 The disulfide bond linkages of eight cysteine residues in the conserved zona domain 1228:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 58 from domain 1305:1310 arg1 linkages 1247:1254 The disulfide bond linkages 1228:1254 The disulfide bond linkages of eight cysteine residues in the conserved zona domain 1228:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 7 59 theme N-terminal 1082:1091 arg1 glutamine 1093:1101 The resultant N-terminal glutamine 1068:1101 The resultant N-terminal glutamine 1068:1101 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 59 theme N-terminal 1082:1091 arg1 to 1116:1117 to 1116:1117 to 1116:1117 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 59 theme N-terminal 1082:1091 arg1 pyrGln 1134:1139 pyrGln(23) 1134:1143 pyrGln(23) 1134:1143 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 59 theme N-terminal 1082:1091 arg1 pyroglutamate 1119:1131 pyroglutamate 1119:1131 pyroglutamate 1119:1131 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 9 60 dep acids 1759:1763 arg1 156-173 1765:1771 156-173 1765:1771 156-173 1765:1771 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 60 dep acids 1759:1763 arg1 260-281 1777:1783 260-281 1777:1783 260-281 1777:1783 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 8 61 theme disulfide 1446:1454 arg1 bonds 1456:1460 two additional disulfide bonds 1431:1460 two additional disulfide bonds 1431:1460 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 1 62 theme human 198:202 arg1 sperm 204:208 human sperm 198:208 human sperm 198:208 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 6 63 theme signal 940:945 arg1 peptide 947:953 an N-terminal signal peptide 926:953 an N-terminal signal peptide (amino acids 1-22) 926:972 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 63 theme signal 940:945 arg1 acids 962:966 amino acids 1-22 956:971 amino acids 1-22 956:971 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 9 64 used occupied 1683:1690 arg2 Three 1580:1584 Three 1580:1584 Three 1580:1584 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 64 used occupied 1683:1690 arg2 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 10 65 theme alternative 1879:1889 arg1 explanations 1891:1902 alternative explanations 1879:1902 alternative explanations of taxon-specific sperm binding 1879:1934 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 9 66 theme amino 1753:1757 arg1 regions 1744:1750 two regions 1740:1750 two regions 1740:1750 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 66 theme amino 1753:1757 arg1 acids 1759:1763 amino acids 156-173 and 260-281 1753:1783 amino acids 156-173 and 260-281 1753:1783 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 67 dep sites 1641:1645 arg1 125 1652:1654 125 1652:1654 125 1652:1654 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 67 dep sites 1641:1645 arg1 Asn 1668:1670 Asn(272) 1668:1675 Asn(272) 1668:1675 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 67 dep sites 1641:1645 arg1 Asn 1658:1660 Asn 1658:1660 Asn(147) 1658:1665 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 67 dep sites 1641:1645 arg1 Asn 1648:1650 Asn 1648:1650 Asn(125) 1648:1655 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 9 68 link N-linked 1608:1615 arg1 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 4 69 theme material 589:596 arg1 paucity 567:573 the paucity 563:573 the paucity of biological material 563:596 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 5 70 theme sperm-egg 632:640 arg1 recognition 642:652 taxon-specific sperm-egg recognition 617:652 taxon-specific sperm-egg recognition 617:652 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 10 71 theme sperm 1922:1926 arg1 binding 1928:1934 taxon-specific sperm binding 1907:1934 taxon-specific sperm binding 1907:1934 Taken together, these data indicate that human and mouse ZP3 proteins are quite similar, and alternative explanations of taxon-specific sperm binding warrant exploration. 15379548 9 72 theme N-linked 1608:1615 arg1 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 1 73 theme extracellular 125:137 arg1 matrix 139:144 an extracellular matrix 122:144 an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs 122:236 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 1 73 theme extracellular 125:137 arg1 pellucida 109:117 The zona pellucida 100:117 The zona pellucida 100:117 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 8 74 theme Cys 1348:1350 arg1 /Cys 1337:1340 Cys(46)/Cys(140) 1330:1345 Cys(46)/Cys(140) 1330:1345 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 74 theme Cys 1348:1350 arg1 /Cys 1355:1358 Cys(78)/Cys(99) 1348:1362 Cys(78)/Cys(99) 1348:1362 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 9 75 theme binding 1633:1639 arg1 sites 1641:1645 the four potential N-linked oligosaccharide binding sites 1589:1645 the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) 1589:1676 Three of the four potential N-linked oligosaccharide binding sites (Asn(125), Asn(147), Asn(272)) were occupied, and clusters of O-glycans were observed within two regions, amino acids 156-173 and 260-281. 15379548 0 76 theme glycosylation-deficient 65:87 arg1 cells 93:97 glycosylation-deficient CHO cells 65:97 glycosylation-deficient CHO cells 65:97 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 2 77 theme zona 249:252 arg1 pellucida 254:262 The mouse zona pellucida 239:262 The mouse zona pellucida 239:262 The mouse zona pellucida is composed of three glycoproteins (ZP1, ZP2, ZP3). 15379548 4 78 dep structure 489:497 arg1 the 475:477 the 475:477 the 475:477 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 8 79 theme residues 1529:1536 arg1 clustering 1487:1496 clustering 1487:1496 clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)) 1487:1577 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 1 80 theme taxon-specific 160:173 arg1 fertilization 175:187 taxon-specific fertilization 160:187 taxon-specific fertilization in which human sperm will not bind to mouse eggs 160:236 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 6 81 dep acids 962:966 arg1 1-22 968:971 1-22 968:971 amino acids 1-22 956:971 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 8 82 theme bond 1242:1245 arg1 linkages 1247:1254 The disulfide bond linkages 1228:1254 The disulfide bond linkages of eight cysteine residues in the conserved zona domain 1228:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 6 83 theme transmembrane 1024:1036 arg1 domain 1038:1043 a C-terminal transmembrane domain 1011:1043 a C-terminal transmembrane domain (amino acids 379-424) 1011:1065 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 83 theme transmembrane 1024:1036 arg1 acids 1052:1056 amino acids 379-424 1046:1064 amino acids 379-424 1046:1064 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 3 84 theme primary 320:326 arg1 structure 328:336 The primary structure 316:336 The primary structure of each 316:344 The primary structure of each has been deduced from the cDNA nucleic acid sequence, and each has been analyzed by mass spectrometry. 15379548 0 85 theme spectrometry 5:16 arg1 analysis 18:25 Mass spectrometry analysis 0:25 Mass spectrometry analysis of recombinant human ZP3 0:50 Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. 15379548 8 86 theme cysteine 1265:1272 arg1 residues 1274:1281 eight cysteine residues 1259:1281 eight cysteine residues in the conserved zona domain 1259:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 5 87 theme CHO-Lec3.2.8.1 783:796 arg1 cells 798:802 CHO-Lec3.2.8.1 cells 783:802 CHO-Lec3.2.8.1 cells 783:802 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 1 88 theme zona 104:107 arg1 matrix 139:144 an extracellular matrix 122:144 an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs 122:236 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 1 88 theme zona 104:107 arg1 pellucida 109:117 The zona pellucida 100:117 The zona pellucida 100:117 The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization in which human sperm will not bind to mouse eggs. 15379548 6 89 theme ZP3 1002:1004 arg1 release 982:988 the release 978:988 the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424) 978:1065 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 6 89 theme ZP3 1002:1004 arg1 cleavage 914:921 the cleavage 910:921 the cleavage of an N-terminal signal peptide (amino acids 1-22) 910:972 With nearly complete coverage, LC-QTOF mass spectrometry was used to determine the cleavage of an N-terminal signal peptide (amino acids 1-22) and the release of secreted ZP3 from a C-terminal transmembrane domain (amino acids 379-424). 15379548 4 90 theme human 521:525 arg1 proteins 532:539 the human zona proteins 517:539 the human zona proteins 517:539 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 5 91 theme recombinant 744:754 arg1 ZP3 762:764 recombinant human ZP3 744:764 recombinant human ZP3 744:764 To investigate if taxon-specific sperm-egg recognition was ascribable to structural differences in a zona protein required for matrix formation, recombinant human ZP3 was expressed in CHO-Lec3.2.8.1 cells and compared to mouse ZP3. 15379548 3 92 theme nucleic 377:383 arg1 sequence 390:397 the cDNA nucleic acid sequence 368:397 the cDNA nucleic acid sequence 368:397 The primary structure of each has been deduced from the cDNA nucleic acid sequence, and each has been analyzed by mass spectrometry. 15379548 4 93 theme secondary 479:487 arg1 structure 489:497 secondary structure 479:497 secondary structure 479:497 However, determination of the secondary structure and processing of the human zona proteins have been hampered by the paucity of biological material. 15379548 8 94 theme conserved 1290:1298 arg1 domain 1305:1310 the conserved zona domain 1286:1310 the conserved zona domain 1286:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 7 95 theme resultant 1072:1080 arg1 glutamine 1093:1101 The resultant N-terminal glutamine 1068:1101 The resultant N-terminal glutamine 1068:1101 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 95 theme resultant 1072:1080 arg1 to 1116:1117 to 1116:1117 to 1116:1117 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 95 theme resultant 1072:1080 arg1 pyrGln 1134:1139 pyrGln(23) 1134:1143 pyrGln(23) 1134:1143 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 7 95 theme resultant 1072:1080 arg1 pyroglutamate 1119:1131 pyroglutamate 1119:1131 pyroglutamate 1119:1131 The resultant N-terminal glutamine was cyclized to pyroglutamate (pyrGln(23)), and several C-terminal peptides were detected, including one ending at Asn(350). 15379548 8 96 dep residues 1529:1536 arg1 Cys 1559:1561 Cys(322) 1559:1566 Cys(322) 1559:1566 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 96 dep residues 1529:1536 arg1 Cys 1549:1551 Cys(321) 1549:1556 Cys(321) 1549:1556 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 96 dep residues 1529:1536 arg1 Cys 1539:1541 Cys(319) 1539:1546 Cys(319) 1539:1546 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 96 dep residues 1529:1536 arg1 Cys 1569:1571 Cys(327) 1569:1576 Cys(327) 1569:1576 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 97 theme Cys 1384:1386 arg1 /Cys 1337:1340 Cys(46)/Cys(140) 1330:1345 Cys(46)/Cys(140) 1330:1345 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 97 theme Cys 1384:1386 arg1 /Cys 1392:1395 Cys(239)/Cys(300) 1384:1400 Cys(239)/Cys(300) 1384:1400 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 3 98 theme mass 430:433 arg1 spectrometry 435:446 mass spectrometry 430:446 mass spectrometry 430:446 The primary structure of each has been deduced from the cDNA nucleic acid sequence, and each has been analyzed by mass spectrometry. 15379548 8 99 from linkages 1247:1254 arg1 domain 1305:1310 the conserved zona domain 1286:1310 the conserved zona domain 1286:1310 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 100 theme Cys 1330:1332 arg1 /Cys 1337:1340 Cys(46)/Cys(140) 1330:1345 Cys(46)/Cys(140) 1330:1345 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 100 theme Cys 1330:1332 arg1 /Cys 1392:1395 Cys(239)/Cys(300) 1384:1400 Cys(239)/Cys(300) 1384:1400 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 100 theme Cys 1330:1332 arg1 /Cys 1355:1358 Cys(78)/Cys(99) 1348:1362 Cys(78)/Cys(99) 1348:1362 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 100 theme Cys 1330:1332 arg1 /Cys 1373:1376 Cys(217)/Cys(282) 1365:1381 Cys(217)/Cys(282) 1365:1381 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 101 theme precise 1412:1418 arg1 indeterminate 1466:1478 indeterminate 1466:1478 indeterminate 1466:1478 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 15379548 8 101 theme precise 1412:1418 arg1 linkage 1420:1426 the precise linkage 1408:1426 the precise linkage of two additional disulfide bonds 1408:1460 The disulfide bond linkages of eight cysteine residues in the conserved zona domain were ascertained (Cys(46)/Cys(140), Cys(78)/Cys(99), Cys(217)/Cys(282), Cys(239)/Cys(300)), but the precise linkage of two additional disulfide bonds was indeterminate due to clustering of the remaining four cysteine residues (Cys(319), Cys(321), Cys(322), Cys(327)). 19004833 0 0 theme N-glycosylation 71:85 arg1 Identification 0:13 Identification 0:13 Identification of the active site of DS-epimerase 1 0:50 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 0 0 theme N-glycosylation 71:85 arg1 requirement 56:66 requirement 56:66 requirement of N-glycosylation for enzyme function 56:105 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 7 1 theme Concomitant 1151:1161 arg1 recreation 1163:1172 Concomitant recreation 1151:1172 Concomitant recreation of the glycosidic linkage 1151:1198 Concomitant recreation of the glycosidic linkage ends the reaction, generating iduronic acid. 19004833 7 2 theme glycosidic 1181:1190 arg1 linkage 1192:1198 the glycosidic linkage 1177:1198 the glycosidic linkage 1177:1198 Concomitant recreation of the glycosidic linkage ends the reaction, generating iduronic acid. 19004833 6 3 from side 1074:1077 arg1 carbon 1047:1052 the C5 carbon 1040:1052 the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction 1040:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 5 4 theme general 847:853 arg1 base 855:858 a general base 845:858 a general base abstracting the C5 proton from glucuronic acid 845:905 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 5 4 theme general 847:853 arg1 His-450 824:830 His-450 824:830 His-450 824:830 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 6 5 theme linkage 946:952 arg1 cleavage 919:926 Subsequent cleavage 908:926 Subsequent cleavage of the glycosidic linkage by Tyr-261 908:963 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 0 6 theme enzyme 91:96 arg1 function 98:105 enzyme function 91:105 enzyme function 91:105 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 3 7 from residues 440:447 arg1 Tyr-261 477:483 Tyr-261 477:483 Tyr-261 477:483 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 3 7 from residues 440:447 arg1 His-450 490:496 His-450 490:496 His-450 490:496 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 3 7 from residues 440:447 arg1 His-205 468:474 His-205 468:474 His-205 468:474 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 3 7 from residues 440:447 arg1 DS-epimerase 452:463 DS-epimerase 1 452:465 DS-epimerase 1 452:465 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 4 8 theme conserved 638:646 arg1 residues 648:655 more conserved residues 633:655 more conserved residues 633:655 These residues were systematically mutated to alanine or more conserved residues, which resulted in complete loss of epimerase activity. 19004833 4 9 theme activity 703:710 arg1 loss 685:688 complete loss 676:688 complete loss of epimerase activity 676:710 These residues were systematically mutated to alanine or more conserved residues, which resulted in complete loss of epimerase activity. 19004833 6 10 theme glycosidic 935:944 arg1 linkage 946:952 the glycosidic linkage 931:952 the glycosidic linkage 931:952 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 6 11 theme opposite 1098:1105 arg1 side 1074:1077 the side 1070:1077 the side of the sugar plane opposite to the side of previous proton abstraction 1070:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 2 12 contain have 327:330 arg2 role 338:341 a key role 332:341 a key role 332:341 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 2 12 contain have 327:330 arg1 domains 245:251 Iduronic acid domains 231:251 Iduronic acid domains 231:251 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 7 13 theme linkage 1192:1198 arg1 recreation 1163:1172 Concomitant recreation 1151:1172 Concomitant recreation of the glycosidic linkage 1151:1198 Concomitant recreation of the glycosidic linkage ends the reaction, generating iduronic acid. 19004833 8 14 theme proper 1271:1276 arg1 N-glycosylation 1278:1292 proper N-glycosylation 1271:1292 proper N-glycosylation of DS-epimerase 1 1271:1310 In addition, we show that proper N-glycosylation of DS-epimerase 1 is required for enzyme activity. 19004833 4 15 theme complete 676:683 arg1 loss 685:688 complete loss 676:688 complete loss of epimerase activity 676:710 These residues were systematically mutated to alanine or more conserved residues, which resulted in complete loss of epimerase activity. 19004833 8 16 theme DS-epimerase 1297:1308 arg1 N-glycosylation 1278:1292 proper N-glycosylation 1271:1292 proper N-glycosylation of DS-epimerase 1 1271:1310 In addition, we show that proper N-glycosylation of DS-epimerase 1 is required for enzyme activity. 19004833 9 17 theme basis 1407:1411 arg1 description 1377:1387 the first description 1367:1387 the first description of the structural basis for epimerization 1367:1429 This study represents the first description of the structural basis for epimerization by a glycosaminoglycan epimerase. 19004833 6 18 theme sugar 1086:1090 arg1 plane 1092:1096 the sugar plane 1082:1096 the sugar plane 1082:1096 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 9 19 theme glycosaminoglycan 1436:1452 arg1 epimerase 1454:1462 a glycosaminoglycan epimerase 1434:1462 a glycosaminoglycan epimerase 1434:1462 This study represents the first description of the structural basis for epimerization by a glycosaminoglycan epimerase. 19004833 6 20 theme C5 1044:1045 arg1 carbon 1047:1052 the C5 carbon 1040:1052 the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction 1040:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 4 21 theme epimerase 693:701 arg1 activity 703:710 epimerase activity 693:710 epimerase activity 693:710 These residues were systematically mutated to alanine or more conserved residues, which resulted in complete loss of epimerase activity. 19004833 6 22 theme plane 1092:1096 arg1 side 1074:1077 the side 1070:1077 the side of the sugar plane opposite to the side of previous proton abstraction 1070:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 2 23 from domains 245:251 arg1 sulfate 265:271 dermatan sulfate 256:271 dermatan sulfate 256:271 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 3 24 theme catalytic 396:404 arg1 site 406:409 the catalytic site 392:409 the catalytic site 392:409 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 0 25 theme active 22:27 arg1 DS-epimerase 37:48 DS-epimerase 1 37:50 DS-epimerase 1 37:50 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 0 25 theme active 22:27 arg1 site 29:32 the active site 18:32 the active site of DS-epimerase 1 18:50 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 8 26 gly N-glycosylation 1278:1292 arg1 enzyme 1328:1333 enzyme activity 1328:1342 enzyme activity 1328:1342 In addition, we show that proper N-glycosylation of DS-epimerase 1 is required for enzyme activity. 19004833 8 26 gly N-glycosylation 1278:1292 arg1 DS-epimerase 1297:1308 DS-epimerase 1 1297:1310 DS-epimerase 1 1297:1310 In addition, we show that proper N-glycosylation of DS-epimerase 1 is required for enzyme activity. 19004833 5 27 theme C5 876:877 arg1 proton 879:884 the C5 proton 872:884 the C5 proton from glucuronic acid 872:905 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 3 28 theme tertiary 502:509 arg1 modeling 521:528 tertiary structure modeling 502:528 tertiary structure modeling 502:528 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 1 29 theme biological 182:191 arg1 functions 193:201 biological functions 182:201 biological functions in development and disease 182:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 5 30 theme epimerase 778:786 arg1 reactions 788:796 epimerase reactions 778:796 epimerase reactions 778:796 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 6 31 theme Subsequent 908:917 arg1 cleavage 919:926 Subsequent cleavage 908:926 Subsequent cleavage of the glycosidic linkage by Tyr-261 908:963 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 3 32 theme structure 511:519 arg1 modeling 521:528 tertiary structure modeling 502:528 tertiary structure modeling 502:528 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 9 33 theme first 1371:1375 arg1 description 1377:1387 the first description 1367:1387 the first description of the structural basis for epimerization 1367:1429 This study represents the first description of the structural basis for epimerization by a glycosaminoglycan epimerase. 19004833 1 34 theme functions 193:201 arg1 variety 171:177 a variety 169:177 a variety of biological functions in development and disease 169:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 34 theme functions 193:201 arg1 functions 193:201 biological functions 182:201 biological functions in development and disease 182:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 7 35 theme iduronic 1230:1237 arg1 acid 1239:1242 iduronic acid 1230:1242 iduronic acid 1230:1242 Concomitant recreation of the glycosidic linkage ends the reaction, generating iduronic acid. 19004833 2 36 theme Iduronic 231:238 arg1 domains 245:251 Iduronic acid domains 231:251 Iduronic acid domains 231:251 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 9 37 theme structural 1396:1405 arg1 basis 1407:1411 the structural basis 1392:1411 the structural basis for epimerization 1392:1429 This study represents the first description of the structural basis for epimerization by a glycosaminoglycan epimerase. 19004833 0 38 theme site 29:32 arg1 Identification 0:13 Identification 0:13 Identification of the active site of DS-epimerase 1 0:50 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 0 38 theme site 29:32 arg1 requirement 56:66 requirement 56:66 requirement of N-glycosylation for enzyme function 56:105 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 8 39 theme enzyme 1328:1333 arg1 activity 1335:1342 enzyme activity 1328:1342 enzyme activity 1328:1342 In addition, we show that proper N-glycosylation of DS-epimerase 1 is required for enzyme activity. 19004833 3 40 theme putative 421:428 arg1 residues 440:447 three putative catalytic residues 415:447 three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450 415:496 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 5 41 theme close 741:745 arg1 relationship 747:758 the close relationship 737:758 the close relationship between lyase and epimerase reactions 737:796 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 1 42 contain has 165:167 arg2 functions 193:201 biological functions 182:201 biological functions in development and disease 182:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 42 contain has 165:167 arg1 polysaccharide 146:159 a highly sulfated polysaccharide 128:159 a highly sulfated polysaccharide 128:159 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 42 contain has 165:167 arg1 sulfate 117:123 Dermatan sulfate 108:123 Dermatan sulfate 108:123 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 42 contain has 165:167 arg2 variety 171:177 a variety 169:177 a variety of biological functions in development and disease 169:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 3 43 theme amino 534:538 arg1 conservation 545:556 amino acid conservation 534:556 amino acid conservation 534:556 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 1 44 from development 206:216 arg1 variety 171:177 a variety 169:177 a variety of biological functions in development and disease 169:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 44 from development 206:216 arg1 functions 193:201 biological functions 182:201 biological functions in development and disease 182:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 45 theme Dermatan 108:115 arg1 polysaccharide 146:159 a highly sulfated polysaccharide 128:159 a highly sulfated polysaccharide 128:159 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 45 theme Dermatan 108:115 arg1 sulfate 117:123 Dermatan sulfate 108:123 Dermatan sulfate 108:123 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 0 46 theme DS-epimerase 37:48 arg1 DS-epimerase 37:48 DS-epimerase 1 37:50 DS-epimerase 1 37:50 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 0 46 theme DS-epimerase 37:48 arg1 site 29:32 the active site 18:32 the active site of DS-epimerase 1 18:50 Identification of the active site of DS-epimerase 1 and requirement of N-glycosylation for enzyme function. 19004833 3 47 theme acid 540:543 arg1 conservation 545:556 amino acid conservation 534:556 amino acid conservation 534:556 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 2 48 theme DS-epimerases 312:324 arg1 action 298:303 the action 294:303 the action of two DS-epimerases 294:324 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 6 49 theme previous 1122:1129 arg1 abstraction 1138:1148 previous proton abstraction 1122:1148 previous proton abstraction 1122:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 2 50 theme dermatan 256:263 arg1 sulfate 265:271 dermatan sulfate 256:271 dermatan sulfate 256:271 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 1 51 from functions 193:201 arg1 disease 222:228 disease 222:228 disease 222:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 51 from functions 193:201 arg1 development 206:216 development 206:216 development 206:216 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 5 52 theme glucuronic 891:900 arg1 acid 902:905 glucuronic acid 891:905 glucuronic acid 891:905 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 6 53 theme 4,5-unsaturated 977:991 arg1 intermediate 1003:1014 a 4,5-unsaturated hexuronic intermediate 975:1014 a 4,5-unsaturated hexuronic intermediate 975:1014 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 1 54 from disease 222:228 arg1 variety 171:177 a variety 169:177 a variety of biological functions in development and disease 169:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 54 from disease 222:228 arg1 functions 193:201 biological functions 182:201 biological functions in development and disease 182:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 6 55 theme abstraction 1138:1148 arg1 side 1114:1117 the side 1110:1117 the side of previous proton abstraction 1110:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 1 56 from variety 171:177 arg1 disease 222:228 disease 222:228 disease 222:228 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 56 from variety 171:177 arg1 development 206:216 development 206:216 development 206:216 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 57 theme sulfated 137:144 arg1 polysaccharide 146:159 a highly sulfated polysaccharide 128:159 a highly sulfated polysaccharide 128:159 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 1 57 theme sulfated 137:144 arg1 sulfate 117:123 Dermatan sulfate 108:123 Dermatan sulfate 108:123 Dermatan sulfate is a highly sulfated polysaccharide and has a variety of biological functions in development and disease. 19004833 6 58 theme hexuronic 993:1001 arg1 intermediate 1003:1014 a 4,5-unsaturated hexuronic intermediate 975:1014 a 4,5-unsaturated hexuronic intermediate 975:1014 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 6 59 theme proton 1131:1136 arg1 abstraction 1138:1148 previous proton abstraction 1122:1148 previous proton abstraction 1122:1148 Subsequent cleavage of the glycosidic linkage by Tyr-261 generates a 4,5-unsaturated hexuronic intermediate, which is protonated at the C5 carbon by His-205 from the side of the sugar plane opposite to the side of previous proton abstraction. 19004833 3 60 theme catalytic 430:438 arg1 residues 440:447 three putative catalytic residues 415:447 three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450 415:496 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 5 61 from acid 902:905 arg1 proton 879:884 the C5 proton 872:884 the C5 proton from glucuronic acid 872:905 Based on these data and the close relationship between lyase and epimerase reactions, we propose a model where His-450 functions as a general base abstracting the C5 proton from glucuronic acid. 19004833 2 62 theme acid 240:243 arg1 domains 245:251 Iduronic acid domains 231:251 Iduronic acid domains 231:251 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 19004833 3 63 from site 406:409 arg1 Tyr-261 477:483 Tyr-261 477:483 Tyr-261 477:483 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 3 63 from site 406:409 arg1 His-450 490:496 His-450 490:496 His-450 490:496 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 3 63 from site 406:409 arg1 His-205 468:474 His-205 468:474 His-205 468:474 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 3 63 from site 406:409 arg1 DS-epimerase 452:463 DS-epimerase 1 452:465 DS-epimerase 1 452:465 We have identified the catalytic site and three putative catalytic residues in DS-epimerase 1, His-205, Tyr-261, and His-450, by tertiary structure modeling and amino acid conservation to heparinase II. 19004833 2 64 theme key 334:336 arg1 role 338:341 a key role 332:341 a key role 332:341 Iduronic acid domains in dermatan sulfate, which are formed by the action of two DS-epimerases, have a key role in mediating these functions. 26783088 9 0 theme Structural 1617:1626 arg1 data 1628:1631 DATABASE: Structural data 1607:1631 DATABASE: Structural data 1607:1631 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 8 1 from framework 1526:1534 arg1 mapping 1452:1458 the mapping 1448:1458 the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function 1448:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 3 2 theme bond 525:528 arg1 likely 542:547 likely 542:547 likely 542:547 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 3 2 theme bond 525:528 arg1 network 530:536 a disulfide bond network 513:536 a disulfide bond network 513:536 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 1 3 theme catalytic 271:279 arg1 domain 281:286 a conserved catalytic domain 259:286 a conserved catalytic domain 259:286 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 4 4 theme Zn 649:650 arg1 ions 656:659 two Zn(2+) ions 645:659 two Zn(2+) ions 645:659 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 8 5 from characterization 1553:1568 arg1 function 1597:1604 aSMase function 1590:1604 aSMase function 1590:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 1 6 theme 3a 213:214 arg1 SMPDL3a 217:223 3a (SMPDL3a) 213:224 3a (SMPDL3a) 213:224 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 6 7 with complex 1079:1085 arg1 product 1096:1102 the product 5'-cytidine monophosphate (CMP) 1092:1134 the product 5'-cytidine monophosphate (CMP) 1092:1134 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 7 with complex 1079:1085 arg1 consistent 1157:1166 consistent 1157:1166 consistent 1157:1166 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 7 with complex 1079:1085 arg1 structure 1139:1147 a structure 1137:1147 a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1137:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 9 8 theme PDB 1654:1656 arg1 database 1658:1665 the PDB database 1650:1665 the PDB database 1650:1665 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 4 9 theme 2+ 652:653 arg1 ions 656:659 two Zn(2+) ions 645:659 two Zn(2+) ions 645:659 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 6 10 theme substrate/product 1216:1232 arg1 modes 1203:1207 several distinct coordination modes 1173:1207 several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1173:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 0 11 with homologue 113:121 arg1 activity 157:164 a novel nucleotide hydrolase activity 128:164 a novel nucleotide hydrolase activity 128:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 9 12 from database 1658:1665 arg1 available 1637:1645 available 1637:1645 available 1637:1645 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 6 13 theme coordination 1190:1201 arg1 modes 1203:1207 several distinct coordination modes 1173:1207 several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1173:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 0 14 theme novel 130:134 arg1 activity 157:164 a novel nucleotide hydrolase activity 128:164 a novel nucleotide hydrolase activity 128:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 5 15 theme recent 779:784 arg1 work 798:801 recent biochemical work 779:801 recent biochemical work 779:801 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 9 16 theme accession 1677:1685 arg1 numbers 1687:1693 the accession numbers 5EBB and 5EBE 1673:1707 the accession numbers 5EBB and 5EBE 1673:1707 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 9 16 theme accession 1677:1685 arg1 5EBE 1704:1707 5EBE 1704:1707 5EBE 1704:1707 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 9 16 theme accession 1677:1685 arg1 5EBB 1695:1698 5EBB 1695:1698 5EBB 1695:1698 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 7 17 theme transfer 1345:1352 arg1 mechanism 1354:1362 a phosphoryl transfer mechanism 1332:1362 a phosphoryl transfer mechanism for SMPDL3a 1332:1374 Based on the structure of CMP complexes, we propose a phosphoryl transfer mechanism for SMPDL3a. 26783088 9 18 theme DATABASE 1607:1614 arg1 data 1628:1631 DATABASE: Structural data 1607:1631 DATABASE: Structural data 1607:1631 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 6 19 from substrate/product 1216:1232 arg1 site 1248:1251 the active site 1237:1251 the active site during the reaction cycle 1237:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 0 20 theme catalytic 18:26 arg1 mechanism 28:36 catalytic mechanism 18:36 catalytic mechanism 18:36 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 5 21 theme work 798:801 arg1 extension 766:774 an extension 763:774 an extension of recent biochemical work 763:801 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 9 22 from available 1637:1645 arg1 database 1658:1665 the PDB database 1650:1665 the PDB database 1650:1665 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 8 23 theme aSMase 1412:1417 arg1 model 1397:1401 a homology model 1386:1401 a homology model of human aSMase 1386:1417 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 5 24 theme modified 865:872 arg1 nucleotides 874:884 several modified nucleotides 857:884 several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin 857:1021 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 6 25 theme distinct 1181:1188 arg1 modes 1203:1207 several distinct coordination modes 1173:1207 several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1173:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 8 26 from function 1597:1604 arg1 characterization 1553:1568 further characterization 1545:1568 further characterization of their effects on aSMase function 1545:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 4 27 used occupied 633:640 arg2 SMPDL3a 622:628 SMPDL3a 622:628 SMPDL3a 622:628 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 4 27 used occupied 633:640 arg2 site 614:617 the binuclear site 600:617 the binuclear site of SMPDL3a 600:628 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 7 28 theme complexes 1310:1318 arg1 structure 1293:1301 the structure 1289:1301 the structure of CMP complexes 1289:1318 Based on the structure of CMP complexes, we propose a phosphoryl transfer mechanism for SMPDL3a. 26783088 0 29 theme phosphodiesterase 61:77 arg1 structure 4:12 structure 4:12 structure 4:12 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 0 29 theme phosphodiesterase 61:77 arg1 mechanism 28:36 catalytic mechanism 18:36 catalytic mechanism 18:36 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 1 30 theme Human 167:171 arg1 enzyme 240:245 a secreted enzyme 229:245 a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease 229:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 30 theme Human 167:171 arg1 phosphodiesterase 190:206 Human sphingomyelinase phosphodiesterase 167:206 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) 167:224 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 0 31 theme 3a--an 84:89 arg1 sphingomyelinase 96:111 3a--an acid sphingomyelinase 84:111 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity 84:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 6 32 with consistent 1157:1166 arg1 modes 1203:1207 several distinct coordination modes 1173:1207 several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1173:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 4 33 theme enzyme 707:712 arg1 activity 714:721 enzyme activity 707:721 enzyme activity 707:721 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 0 34 theme sphingomyelinase 96:111 arg1 homologue 113:121 3a--an acid sphingomyelinase homologue 84:121 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity 84:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 5 35 theme diphosphate 938:948 arg1 ethanolamine 950:961 cytidine diphosphate ethanolamine 929:961 cytidine diphosphate ethanolamine 929:961 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 8 36 theme Niemann-Pick 1472:1483 arg1 disease 1485:1491 Niemann-Pick disease 1472:1491 selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function 1463:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 1 37 theme disease 387:393 arg1 causative 361:369 causative 361:369 causative 361:369 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 7 38 theme CMP 1306:1308 arg1 complexes 1310:1318 CMP complexes 1306:1318 CMP complexes 1306:1318 Based on the structure of CMP complexes, we propose a phosphoryl transfer mechanism for SMPDL3a. 26783088 8 39 theme mutations 1493:1501 arg1 mapping 1452:1458 the mapping 1448:1458 the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function 1448:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 1 40 theme human 293:297 arg1 sphingomyelinase 304:319 human acid sphingomyelinase 293:319 human acid sphingomyelinase (aSMase) 293:328 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 40 theme human 293:297 arg1 enzyme 335:340 the enzyme 331:340 the enzyme carrying mutations causative of Niemann-Pick disease 331:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 40 theme human 293:297 arg1 aSMase 322:327 aSMase 322:327 aSMase 322:327 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 5 41 theme cytidine 899:906 arg1 5'-diphosphocholine 908:926 cytidine 5'-diphosphocholine 899:926 cytidine 5'-diphosphocholine 899:926 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 2 42 theme calcineurin-like 448:463 arg1 fold 465:468 a calcineurin-like fold 446:468 a calcineurin-like fold 446:468 We have solved the structure of SMPDL3a revealing a calcineurin-like fold. 26783088 8 43 from mutations 1493:1501 arg1 framework 1526:1534 a three-dimensional framework 1506:1534 a three-dimensional framework to guide further characterization of their effects on aSMase function 1506:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 5 44 theme aSMase 991:996 arg1 sphingomyelin 1009:1021 sphingomyelin 1009:1021 sphingomyelin 1009:1021 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 5 44 theme aSMase 991:996 arg1 substrate 998:1006 the aSMase substrate 987:1006 the aSMase substrate 987:1006 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 9 45 dep numbers 1687:1693 arg1 numbers 1687:1693 the accession numbers 5EBB and 5EBE 1673:1707 the accession numbers 5EBB and 5EBE 1673:1707 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 9 45 dep numbers 1687:1693 arg1 5EBE 1704:1707 5EBE 1704:1707 5EBE 1704:1707 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 9 45 dep numbers 1687:1693 arg1 5EBB 1695:1698 5EBB 1695:1698 5EBB 1695:1698 DATABASE: Structural data are available in the PDB database under the accession numbers 5EBB and 5EBE. 26783088 0 46 theme human 41:45 arg1 phosphodiesterase 61:77 human sphingomyelin phosphodiesterase 41:77 human sphingomyelin phosphodiesterase 41:77 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 6 47 theme 5'-cytidine 1104:1114 arg1 CMP 1131:1133 CMP 1131:1133 CMP 1131:1133 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 47 theme 5'-cytidine 1104:1114 arg1 monophosphate 1116:1128 5'-cytidine monophosphate 1104:1128 the product 5'-cytidine monophosphate (CMP) 1092:1134 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 48 dep product 1096:1102 arg1 CMP 1131:1133 CMP 1131:1133 CMP 1131:1133 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 48 dep product 1096:1102 arg1 monophosphate 1116:1128 5'-cytidine monophosphate 1104:1128 the product 5'-cytidine monophosphate (CMP) 1092:1134 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 1 49 contain carrying 342:349 arg1 sphingomyelinase 304:319 human acid sphingomyelinase 293:319 human acid sphingomyelinase (aSMase) 293:328 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 49 contain carrying 342:349 arg1 enzyme 335:340 the enzyme 331:340 the enzyme carrying mutations causative of Niemann-Pick disease 331:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 49 contain carrying 342:349 arg2 mutations 351:359 mutations 351:359 mutations causative of Niemann-Pick disease 351:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 6 50 from SMPDL3a 1068:1074 arg1 complex 1079:1085 complex 1079:1085 complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1079:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 51 theme reaction 1264:1271 arg1 cycle 1273:1277 the reaction cycle 1260:1277 the reaction cycle 1260:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 52 from complex 1079:1085 arg1 structure 1055:1063 the structure 1051:1063 the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1051:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 3 53 theme disulfide 515:523 arg1 likely 542:547 likely 542:547 likely 542:547 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 3 53 theme disulfide 515:523 arg1 network 530:536 a disulfide bond network 513:536 a disulfide bond network 513:536 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 1 54 theme conserved 261:269 arg1 domain 281:286 a conserved catalytic domain 259:286 a conserved catalytic domain 259:286 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 6 55 theme SMPDL3a 1068:1074 arg1 structure 1055:1063 the structure 1051:1063 the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1051:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 56 theme active 1241:1246 arg1 site 1248:1251 the active site 1237:1251 the active site during the reaction cycle 1237:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 6 57 from structure 1055:1063 arg1 complex 1079:1085 complex 1079:1085 complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1079:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 3 58 theme human 573:577 arg1 aSMase 579:584 human aSMase 573:584 human aSMase 573:584 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 0 59 theme nucleotide 136:145 arg1 activity 157:164 a novel nucleotide hydrolase activity 128:164 a novel nucleotide hydrolase activity 128:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 8 60 theme human 1406:1410 arg1 aSMase 1412:1417 human aSMase 1406:1417 human aSMase 1406:1417 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 4 61 theme SMPDL3a 622:628 arg1 SMPDL3a 622:628 SMPDL3a 622:628 SMPDL3a 622:628 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 4 61 theme SMPDL3a 622:628 arg1 site 614:617 the binuclear site 600:617 the binuclear site of SMPDL3a 600:628 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 8 62 from effects 1579:1585 arg1 function 1597:1604 aSMase function 1590:1604 aSMase function 1590:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 5 63 theme biochemical 786:796 arg1 work 798:801 recent biochemical work 779:801 recent biochemical work 779:801 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 5 64 theme several 857:863 arg1 nucleotides 874:884 several modified nucleotides 857:884 several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin 857:1021 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 7 65 theme phosphoryl 1334:1343 arg1 mechanism 1354:1362 a phosphoryl transfer mechanism 1332:1362 a phosphoryl transfer mechanism for SMPDL3a 1332:1374 Based on the structure of CMP complexes, we propose a phosphoryl transfer mechanism for SMPDL3a. 26783088 4 66 theme excess 670:675 arg1 Zn 677:678 excess Zn 670:678 excess Zn(2+) 670:682 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 4 66 theme excess 670:675 arg1 2+ 680:681 2+ 680:681 2+ 680:681 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 6 67 from modes 1203:1207 arg1 site 1248:1251 the active site 1237:1251 the active site during the reaction cycle 1237:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 5 68 theme nucleotides 874:884 arg1 hydrolysis 843:852 the hydrolysis 839:852 the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin 839:1021 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 6 69 theme several 1173:1179 arg1 modes 1203:1207 several distinct coordination modes 1173:1207 several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1173:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 3 70 theme glycosylation 487:499 arg1 pattern 501:507 glycosylation pattern 487:507 glycosylation pattern 487:507 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 8 71 theme effects 1579:1585 arg1 characterization 1553:1568 further characterization 1545:1568 further characterization of their effects on aSMase function 1545:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 0 72 theme acid 91:94 arg1 sphingomyelinase 96:111 3a--an acid sphingomyelinase 84:111 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity 84:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 1 73 theme sphingomyelinase 173:188 arg1 enzyme 240:245 a secreted enzyme 229:245 a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease 229:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 73 theme sphingomyelinase 173:188 arg1 phosphodiesterase 190:206 Human sphingomyelinase phosphodiesterase 167:206 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) 167:224 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 8 74 theme selected 1463:1470 arg1 mutations 1493:1501 selected Niemann-Pick disease mutations 1463:1501 selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function 1463:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 1 75 theme causative 361:369 arg1 mutations 351:359 mutations 351:359 mutations causative of Niemann-Pick disease 351:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 5 76 theme cytidine 929:936 arg1 ethanolamine 950:961 cytidine diphosphate ethanolamine 929:961 cytidine diphosphate ethanolamine 929:961 As an extension of recent biochemical work we uncovered that SMPDL3a catalyses the hydrolysis of several modified nucleotides that include cytidine 5'-diphosphocholine, cytidine diphosphate ethanolamine and ADP-ribose, but not the aSMase substrate, sphingomyelin. 26783088 8 77 theme disease 1485:1491 arg1 mutations 1493:1501 selected Niemann-Pick disease mutations 1463:1501 selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function 1463:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 1 78 theme Niemann-Pick 374:385 arg1 disease 387:393 Niemann-Pick disease 374:393 Niemann-Pick disease 374:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 4 79 theme activity 714:721 arg1 inhibition 693:702 inhibition 693:702 inhibition of enzyme activity 693:721 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 8 80 theme three-dimensional 1508:1524 arg1 framework 1526:1534 a three-dimensional framework 1506:1534 a three-dimensional framework to guide further characterization of their effects on aSMase function 1506:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 8 81 from mapping 1452:1458 arg1 framework 1526:1534 a three-dimensional framework 1506:1534 a three-dimensional framework to guide further characterization of their effects on aSMase function 1506:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 1 82 theme acid 299:302 arg1 sphingomyelinase 304:319 human acid sphingomyelinase 293:319 human acid sphingomyelinase (aSMase) 293:328 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 82 theme acid 299:302 arg1 enzyme 335:340 the enzyme 331:340 the enzyme carrying mutations causative of Niemann-Pick disease 331:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 82 theme acid 299:302 arg1 aSMase 322:327 aSMase 322:327 aSMase 322:327 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 2 83 theme SMPDL3a 428:434 arg1 structure 415:423 the structure 411:423 the structure of SMPDL3a 411:434 We have solved the structure of SMPDL3a revealing a calcineurin-like fold. 26783088 8 84 theme further 1545:1551 arg1 characterization 1553:1568 further characterization 1545:1568 further characterization of their effects on aSMase function 1545:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 3 85 theme dimetal 473:479 arg1 likely 542:547 likely 542:547 likely 542:547 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 3 85 theme dimetal 473:479 arg1 site 481:484 A dimetal site 471:484 A dimetal site 471:484 A dimetal site, glycosylation pattern and a disulfide bond network are likely to be conserved also in human aSMase. 26783088 0 86 theme sphingomyelin 47:59 arg1 phosphodiesterase 61:77 human sphingomyelin phosphodiesterase 41:77 human sphingomyelin phosphodiesterase 41:77 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 0 87 theme hydrolase 147:155 arg1 activity 157:164 a novel nucleotide hydrolase activity 128:164 a novel nucleotide hydrolase activity 128:164 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 4 88 theme additional 742:751 arg1 sites 753:757 additional sites 742:757 additional sites 742:757 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 4 89 theme binuclear 604:612 arg1 SMPDL3a 622:628 SMPDL3a 622:628 SMPDL3a 622:628 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 4 89 theme binuclear 604:612 arg1 site 614:617 the binuclear site 600:617 the binuclear site of SMPDL3a 600:628 We show that the binuclear site of SMPDL3a is occupied by two Zn(2+) ions and that excess Zn(2+) leads to inhibition of enzyme activity through binding to additional sites. 26783088 6 90 from site 1248:1251 arg1 modes 1203:1207 several distinct coordination modes 1173:1207 several distinct coordination modes of the substrate/product in the active site during the reaction cycle 1173:1277 We subsequently determined the structure of SMPDL3a in complex with the product 5'-cytidine monophosphate (CMP), a structure that is consistent with several distinct coordination modes of the substrate/product in the active site during the reaction cycle. 26783088 1 91 theme secreted 231:238 arg1 enzyme 240:245 a secreted enzyme 229:245 a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease 229:393 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 1 91 theme secreted 231:238 arg1 phosphodiesterase 190:206 Human sphingomyelinase phosphodiesterase 167:206 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) 167:224 Human sphingomyelinase phosphodiesterase like 3a (SMPDL3a) is a secreted enzyme that shares a conserved catalytic domain with human acid sphingomyelinase (aSMase), the enzyme carrying mutations causative of Niemann-Pick disease. 26783088 0 92 dep structure 4:12 arg1 The 0:2 The 0:2 The 0:2 The structure and catalytic mechanism of human sphingomyelin phosphodiesterase like 3a--an acid sphingomyelinase homologue with a novel nucleotide hydrolase activity. 26783088 8 93 theme aSMase 1590:1595 arg1 function 1597:1604 aSMase function 1590:1604 aSMase function 1590:1604 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 26783088 8 94 theme homology 1388:1395 arg1 model 1397:1401 a homology model 1386:1401 a homology model of human aSMase 1386:1417 Finally, a homology model of human aSMase was constructed to allow for the mapping of selected Niemann-Pick disease mutations on a three-dimensional framework to guide further characterization of their effects on aSMase function. 19571171 5 0 theme substrates 860:869 arg1 sulfation 833:841 sulfation 833:841 sulfation of two different substrates 833:869 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 1 theme N410 798:801 arg1 glycans 803:809 The N196 and N410 glycans 785:809 The N196 and N410 glycans 785:809 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 3 2 theme sites 517:521 arg1 three 470:474 three 470:474 three 470:474 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 3 2 theme sites 517:521 arg1 sites 517:521 four potential N-linked glycosylation sites 479:521 four potential N-linked glycosylation sites 479:521 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 5 3 theme epitope 1048:1054 arg1 levels 995:1000 reduced levels 987:1000 reduced levels of a sulfated peripheral lymph node addressin epitope 987:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 4 theme sulfated 1135:1142 arg1 addressin 1166:1174 a sulfated peripheral lymph node addressin 1133:1174 a sulfated peripheral lymph node addressin epitope 1133:1182 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 8 5 theme misglycosylated 1592:1606 arg1 enzyme 1608:1613 misglycosylated enzyme 1592:1613 misglycosylated enzyme 1592:1613 Altered trafficking of mutants may be associated with the mechanisms by which misglycosylated enzyme is degraded. 19571171 5 6 theme peripheral 1144:1153 arg1 addressin 1166:1174 a sulfated peripheral lymph node addressin 1133:1174 a sulfated peripheral lymph node addressin epitope 1133:1182 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 0 7 from Effects 0:6 arg1 activity 34:41 activity 34:41 activity 34:41 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 0 7 from Effects 0:6 arg1 localization 47:58 localization 47:58 localization 47:58 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 0 8 gly N-glycosylation 11:25 arg1 GlcNAc-6-sulfotransferase 63:87 GlcNAc-6-sulfotransferase 1 63:89 GlcNAc-6-sulfotransferase 1 63:89 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 6 9 theme enzyme 1326:1331 arg1 localization 1333:1344 enzyme localization 1326:1344 enzyme localization 1326:1344 The glycans' effects on enzyme activity may be mediated, in part, by changes in enzyme localization. 19571171 5 10 theme lymph 1155:1159 arg1 addressin 1166:1174 a sulfated peripheral lymph node addressin 1133:1174 a sulfated peripheral lymph node addressin epitope 1133:1182 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 4 11 theme surface 610:616 arg1 glycans 618:624 sulfated cell surface glycans 596:624 sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan 596:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 8 12 theme mutants 1537:1543 arg1 trafficking 1522:1532 Altered trafficking 1514:1532 Altered trafficking of mutants 1514:1543 Altered trafficking of mutants may be associated with the mechanisms by which misglycosylated enzyme is degraded. 19571171 4 13 theme sulfate 695:701 arg1 Lewis 714:718 sialyl Lewis 707:718 sialyl Lewis 707:718 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 2 14 contain has 375:377 arg2 effects 350:356 the effects 346:356 the effects that each glycan has on enzyme activity, specificity, and localization 346:427 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 2 14 contain has 375:377 arg1 glycan 368:373 each glycan 363:373 each glycan 363:373 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 2 15 mod modified 311:318 arg1 GlcNAc6ST-1 296:306 GlcNAc6ST-1 296:306 GlcNAc6ST-1 296:306 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 2 15 mod modified 311:318 arg3 glycans 334:340 N-linked glycans 325:340 N-linked glycans 325:340 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 4 16 theme extended 752:759 arg1 glycan 777:782 a putative extended core 1 O-linked glycan 741:782 a putative extended core 1 O-linked glycan 741:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 5 17 theme node 1161:1164 arg1 addressin 1166:1174 a sulfated peripheral lymph node addressin 1133:1174 a sulfated peripheral lymph node addressin epitope 1133:1182 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 7 18 theme most 1353:1356 arg1 mutants 1358:1364 most mutants 1353:1364 most mutants that lacked glycans 1353:1384 While most mutants that lacked glycans localized normally within the Golgi, the N428A mutant and a mutant lacking all glycans were also found to localize ectopically. 19571171 6 19 theme enzyme 1270:1275 arg1 activity 1277:1284 enzyme activity 1270:1284 enzyme activity 1270:1284 The glycans' effects on enzyme activity may be mediated, in part, by changes in enzyme localization. 19571171 4 20 theme putative 743:750 arg1 glycan 777:782 a putative extended core 1 O-linked glycan 741:782 a putative extended core 1 O-linked glycan 741:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 6 21 from effects 1259:1265 arg1 activity 1277:1284 enzyme activity 1270:1284 enzyme activity 1270:1284 The glycans' effects on enzyme activity may be mediated, in part, by changes in enzyme localization. 19571171 3 22 gly glycosylation 503:515 arg2 four 479:482 four 479:482 four 479:482 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 3 22 gly glycosylation 503:515 arg2 sites 517:521 four potential N-linked glycosylation sites 479:521 four potential N-linked glycosylation sites 479:521 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 6 23 from changes 1315:1321 arg1 localization 1333:1344 enzyme localization 1326:1344 enzyme localization 1326:1344 The glycans' effects on enzyme activity may be mediated, in part, by changes in enzyme localization. 19571171 4 24 theme sialyl 707:712 arg1 Lewis 714:718 sialyl Lewis 707:718 sialyl Lewis 707:718 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 5 25 theme Lewis 1237:1241 arg1 X 1243:1243 sulfate sialyl Lewis X 1222:1243 sulfate sialyl Lewis X 1222:1243 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 1 26 theme l-selectin 221:230 arg1 ligand 232:237 the l-selectin ligand 217:237 the l-selectin ligand on endothelial cells 217:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 5 27 theme N196 789:792 arg1 glycans 803:809 The N196 and N410 glycans 785:809 The N196 and N410 glycans 785:809 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 28 dep able 929:932 arg1 cells 872:876 cells 872:876 cells that express an enzyme lacking the N410 glycan 872:923 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 28 dep able 929:932 arg1 substrate 964:972 substrate 964:972 substrate 964:972 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 28 dep able 929:932 arg1 Lewis 956:960 the sialyl Lewis 945:960 the sialyl Lewis 945:960 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 2 29 theme N-linked 325:332 arg1 glycans 334:340 N-linked glycans 325:340 N-linked glycans 325:340 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 1 30 theme ligand 232:237 arg1 sulfation 204:212 sulfation 204:212 sulfation of the l-selectin ligand on endothelial cells 204:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 0 31 theme N-glycosylation 11:25 arg1 Effects 0:6 Effects 0:6 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1 0:89 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 5 32 theme N410 913:916 arg1 glycan 918:923 the N410 glycan 909:923 the N410 glycan 909:923 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 33 theme peripheral 1016:1025 arg1 addressin 1038:1046 a sulfated peripheral lymph node addressin 1005:1046 a sulfated peripheral lymph node addressin epitope 1005:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 4 34 link O-linked 768:775 arg1 glycan 777:782 a putative extended core 1 O-linked glycan 741:782 a putative extended core 1 O-linked glycan 741:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 4 35 dep glycans 618:624 arg1 unable 685:690 unable 685:690 unable 685:690 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 4 36 theme N428 550:553 arg1 glycan 555:560 The N428 glycan 546:560 The N428 glycan 546:560 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 1 37 gly glycoprotein 167:178 arg1 glycoprotein 167:178 a Golgi-resident glycoprotein 150:178 a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells 150:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 1 37 gly glycoprotein 167:178 arg1 responsible 188:198 responsible 188:198 responsible 188:198 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 1 37 gly glycoprotein 167:178 arg1 N-Acetylglucosamine-6-sulfotransferase-1 92:131 N-Acetylglucosamine-6-sulfotransferase-1 92:131 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) 92:145 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 5 38 theme lymph 1027:1031 arg1 addressin 1038:1046 a sulfated peripheral lymph node addressin 1005:1046 a sulfated peripheral lymph node addressin epitope 1005:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 4 39 theme mutant 646:651 arg1 enzyme 653:658 a mutant enzyme 644:658 a mutant enzyme lacking this glycan 644:678 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 1 40 theme endothelial 242:252 arg1 cells 254:258 endothelial cells 242:258 endothelial cells 242:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 2 41 link N-linked 325:332 arg1 glycans 334:340 N-linked glycans 325:340 N-linked glycans 325:340 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 5 42 theme node 1033:1036 arg1 addressin 1038:1046 a sulfated peripheral lymph node addressin 1005:1046 a sulfated peripheral lymph node addressin epitope 1005:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 8 43 theme Altered 1514:1520 arg1 trafficking 1522:1532 Altered trafficking 1514:1532 Altered trafficking of mutants 1514:1543 Altered trafficking of mutants may be associated with the mechanisms by which misglycosylated enzyme is degraded. 19571171 5 44 from ability 1211:1217 arg1 impaired 1193:1200 impaired 1193:1200 impaired 1193:1200 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 45 theme N196 1101:1104 arg1 glycan 1106:1111 the N196 glycan 1097:1111 the N196 glycan 1097:1111 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 46 theme addressin 1038:1046 arg1 epitope 1048:1054 a sulfated peripheral lymph node addressin epitope 1005:1054 a sulfated peripheral lymph node addressin epitope 1005:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 0 47 dep activity 34:41 arg1 the 30:32 the 30:32 the 30:32 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 1 48 from cells 254:258 arg1 sulfation 204:212 sulfation 204:212 sulfation of the l-selectin ligand on endothelial cells 204:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 3 49 theme N-linked 494:501 arg1 sites 517:521 four potential N-linked glycosylation sites 479:521 four potential N-linked glycosylation sites 479:521 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 5 50 theme reduced 987:993 arg1 levels 995:1000 reduced levels 987:1000 reduced levels of a sulfated peripheral lymph node addressin epitope 987:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 1 51 from ligand 232:237 arg1 cells 254:258 endothelial cells 242:258 endothelial cells 242:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 3 52 theme glycosylation 503:515 arg1 sites 517:521 four potential N-linked glycosylation sites 479:521 four potential N-linked glycosylation sites 479:521 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 3 53 link N-linked 494:501 arg1 sites 517:521 four potential N-linked glycosylation sites 479:521 four potential N-linked glycosylation sites 479:521 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 5 54 theme sialyl 949:954 arg1 cells 872:876 cells 872:876 cells that express an enzyme lacking the N410 glycan 872:923 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 54 theme sialyl 949:954 arg1 Lewis 956:960 the sialyl Lewis 945:960 the sialyl Lewis 945:960 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 1 55 theme Golgi-resident 152:165 arg1 glycoprotein 167:178 a Golgi-resident glycoprotein 150:178 a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells 150:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 1 55 theme Golgi-resident 152:165 arg1 responsible 188:198 responsible 188:198 responsible 188:198 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 1 55 theme Golgi-resident 152:165 arg1 N-Acetylglucosamine-6-sulfotransferase-1 92:131 N-Acetylglucosamine-6-sulfotransferase-1 92:131 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) 92:145 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 5 56 theme different 850:858 arg1 substrates 860:869 two different substrates 846:869 two different substrates 846:869 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 57 theme addressin 1166:1174 arg1 epitope 1176:1182 a sulfated peripheral lymph node addressin epitope 1133:1182 a sulfated peripheral lymph node addressin epitope 1133:1182 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 5 58 from impaired 1193:1200 arg1 ability 1211:1217 their ability 1205:1217 their ability 1205:1217 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 7 59 theme N428A 1427:1431 arg1 mutant 1433:1438 the N428A mutant 1423:1438 the N428A mutant 1423:1438 While most mutants that lacked glycans localized normally within the Golgi, the N428A mutant and a mutant lacking all glycans were also found to localize ectopically. 19571171 4 60 theme core 761:764 arg1 glycan 777:782 a putative extended core 1 O-linked glycan 741:782 a putative extended core 1 O-linked glycan 741:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 3 61 theme potential 484:492 arg1 sites 517:521 four potential N-linked glycosylation sites 479:521 four potential N-linked glycosylation sites 479:521 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 5 62 theme sulfate 1222:1228 arg1 X 1243:1243 sulfate sialyl Lewis X 1222:1243 sulfate sialyl Lewis X 1222:1243 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 4 63 dep Lewis 714:718 arg1 the 703:705 the 703:705 the 703:705 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 0 64 theme GlcNAc-6-sulfotransferase 63:87 arg1 activity 34:41 activity 34:41 activity 34:41 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 0 64 theme GlcNAc-6-sulfotransferase 63:87 arg1 localization 47:58 localization 47:58 localization 47:58 Effects of N-glycosylation on the activity and localization of GlcNAc-6-sulfotransferase 1. 19571171 5 65 theme sulfated 1007:1014 arg1 addressin 1038:1046 a sulfated peripheral lymph node addressin 1005:1046 a sulfated peripheral lymph node addressin epitope 1005:1054 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 3 66 dep three 470:474 arg1 N410 530:533 N410 530:533 N410 530:533 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 3 66 dep three 470:474 arg1 N428 540:543 N428 540:543 N428 540:543 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 3 66 dep three 470:474 arg1 N196 524:527 N196 524:527 N196 524:527 We determined that glycans are added at three of four potential N-linked glycosylation sites: N196, N410, and N428. 19571171 4 67 theme glycans 618:624 arg1 production 582:591 the production 578:591 the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan 578:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 2 68 theme enzyme 382:387 arg1 activity 389:396 enzyme activity 382:396 enzyme activity 382:396 Here, we report the sites at which GlcNAc6ST-1 is modified with N-linked glycans and the effects that each glycan has on enzyme activity, specificity, and localization. 19571171 5 69 theme sialyl 1230:1235 arg1 X 1243:1243 sulfate sialyl Lewis X 1222:1243 sulfate sialyl Lewis X 1222:1243 The N196 and N410 glycans differentially affect sulfation of two different substrates: cells that express an enzyme lacking the N410 glycan are able to sulfate the sialyl Lewis X substrate, but produce reduced levels of a sulfated peripheral lymph node addressin epitope and cells that express an enzyme lacking the N196 glycan are able to produce a sulfated peripheral lymph node addressin epitope, but are impaired in their ability to sulfate sialyl Lewis X. 19571171 4 70 theme cell 605:608 arg1 glycans 618:624 sulfated cell surface glycans 596:624 sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan 596:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 8 71 gly misglycosylated 1592:1606 arg1 enzyme 1608:1613 misglycosylated enzyme 1592:1613 misglycosylated enzyme 1592:1613 Altered trafficking of mutants may be associated with the mechanisms by which misglycosylated enzyme is degraded. 19571171 4 72 theme O-linked 768:775 arg1 glycan 777:782 a putative extended core 1 O-linked glycan 741:782 a putative extended core 1 O-linked glycan 741:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 19571171 1 73 from sulfation 204:212 arg1 cells 254:258 endothelial cells 242:258 endothelial cells 242:258 N-Acetylglucosamine-6-sulfotransferase-1 (GlcNAc6ST-1) is a Golgi-resident glycoprotein that is responsible for sulfation of the l-selectin ligand on endothelial cells. 19571171 4 74 theme sulfated 596:603 arg1 glycans 618:624 sulfated cell surface glycans 596:624 sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan 596:782 The N428 glycan is required for the production of sulfated cell surface glycans: cells expressing a mutant enzyme lacking this glycan were unable to sulfate the sialyl Lewis X tetrasaccharide or a putative extended core 1 O-linked glycan. 18405659 8 0 theme HeLa 1388:1391 arg1 cells 1393:1397 HeLa cells 1388:1397 HeLa cells 1388:1397 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 1 1 theme human 144:148 arg1 transporter 172:182 The human proton-coupled folate transporter 140:182 The human proton-coupled folate transporter (HsPCFT, SLC46A1) 140:200 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 8 2 theme amino 1308:1312 arg1 termini 1326:1332 amino or carboxyl termini 1308:1332 termini 1326:1332 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 2 3 theme cell 405:408 arg1 lines 410:414 cell lines 405:414 cell lines 405:414 On Western blot, HsPCFT migrates as a broad band (~55 kDa), higher than predicted (~50 kDa) in cell lines. 18405659 0 4 from glycosylation 9:21 arg1 function 74:81 function 74:81 function 74:81 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 4 from glycosylation 9:21 arg1 mobility 61:68 electrophoretic mobility 45:68 electrophoretic mobility 45:68 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 1 5 theme proton-coupled 150:163 arg1 transporter 172:182 The human proton-coupled folate transporter 140:182 The human proton-coupled folate transporter (HsPCFT, SLC46A1) 140:200 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 0 6 theme human 90:94 arg1 HsPCFT 131:136 HsPCFT 131:136 HsPCFT 131:136 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 6 theme human 90:94 arg1 transporter 118:128 the human proton-coupled folate transporter 86:128 the human proton-coupled folate transporter (HsPCFT) 86:137 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 8 7 theme domains 1477:1483 arg1 location 1459:1466 the intracellular location 1441:1466 the intracellular location of these domains 1441:1483 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 7 8 dep NH 1147:1148 arg1 the 1143:1145 the 1143:1145 the 1143:1145 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 7 9 theme double 1089:1094 arg1 mutant 1096:1101 the double mutant 1085:1101 the double mutant 1085:1101 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 4 10 with cells 631:635 arg1 F 663:663 peptidyl N-glycanase F 642:663 peptidyl N-glycanase F 642:663 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 5 11 gly glycosylation 791:803 arg2 sites 805:809 two canonical glycosylation sites 777:809 two canonical glycosylation sites 777:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 5 11 gly glycosylation 791:803 arg2 two 777:779 two 777:779 two 777:779 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 3 12 theme blot 425:428 arg1 analysis 430:437 Western blot analysis 417:437 Western blot analysis 417:437 Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. 18405659 1 13 theme folates 261:267 arg1 transport 248:256 transport 248:256 transport of folates into the liver, brain and other tissues 248:307 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 1 13 theme folates 261:267 arg1 absorption 222:231 intestinal absorption 211:231 intestinal absorption of folates 211:242 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 0 14 theme folate 111:116 arg1 HsPCFT 131:136 HsPCFT 131:136 HsPCFT 131:136 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 14 theme folate 111:116 arg1 transporter 118:128 the human proton-coupled folate transporter 86:128 the human proton-coupled folate transporter (HsPCFT) 86:137 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 8 15 theme deglycosylated 1272:1285 arg1 HA 1294:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 0 16 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 3 17 theme membrane 453:460 arg1 preparations 462:473 membrane preparations 453:473 membrane preparations 453:473 Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. 18405659 4 18 theme fractions 592:600 arg1 Treatment 570:578 Treatment 570:578 Treatment 570:578 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 1 19 theme folate 165:170 arg1 transporter 172:182 The human proton-coupled folate transporter 140:182 The human proton-coupled folate transporter (HsPCFT, SLC46A1) 140:200 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 0 20 theme proton-coupled 96:109 arg1 HsPCFT 131:136 HsPCFT 131:136 HsPCFT 131:136 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 20 theme proton-coupled 96:109 arg1 transporter 118:128 the human proton-coupled folate transporter 86:128 the human proton-coupled folate transporter (HsPCFT) 86:137 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 7 21 theme Transport 1027:1035 arg1 function 1037:1044 Transport function 1027:1044 Transport function 1027:1044 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 6 22 theme double 980:985 arg1 mutant 987:992 the double mutant 976:992 the double mutant 976:992 Single mutants retained full transport activity; the double mutant retained a majority of activity. 18405659 5 23 theme canonical 781:789 arg1 sites 805:809 two canonical glycosylation sites 777:809 two canonical glycosylation sites 777:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 0 24 from impact 31:36 arg1 function 74:81 function 74:81 function 74:81 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 24 from impact 31:36 arg1 mobility 61:68 electrophoretic mobility 45:68 electrophoretic mobility 45:68 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 8 25 theme intracellular 1445:1457 arg1 location 1459:1466 the intracellular location 1441:1466 the intracellular location of these domains 1441:1483 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 4 26 from cells 669:673 arg1 Treatment 570:578 Treatment 570:578 Treatment 570:578 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 4 26 from cells 669:673 arg1 fractions 592:600 membrane fractions 583:600 membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells 583:673 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 5 27 theme kDa 853:855 arg1 protein 857:863 a ~47 kDa protein 847:863 a ~47 kDa protein 847:863 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 4 28 from Treatment 570:578 arg1 cells 669:673 cells 669:673 cells 669:673 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 4 28 from Treatment 570:578 arg1 cells 631:635 HsPCFT-transfected HeLa cells 607:635 HsPCFT-transfected HeLa cells 607:635 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 2 29 theme broad 348:352 arg1 band 354:357 a broad band 346:357 a broad band (~55 kDa) 346:367 On Western blot, HsPCFT migrates as a broad band (~55 kDa), higher than predicted (~50 kDa) in cell lines. 18405659 2 29 theme broad 348:352 arg1 kDa 364:366 ~55 kDa 360:366 ~55 kDa 360:366 On Western blot, HsPCFT migrates as a broad band (~55 kDa), higher than predicted (~50 kDa) in cell lines. 18405659 0 30 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 3 31 theme loading 499:505 arg1 buffer 507:512 the loading buffer 495:512 the loading buffer 495:512 Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. 18405659 4 32 theme membrane 583:590 arg1 fractions 592:600 membrane fractions 583:600 membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells 583:673 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 8 33 theme carboxyl 1317:1324 arg1 termini 1326:1332 amino or carboxyl termini 1308:1332 termini 1326:1332 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 5 34 theme ~47 849:851 arg1 protein 857:863 a ~47 kDa protein 847:863 a ~47 kDa protein 847:863 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 7 35 dep tagged 1126:1131 arg1 HA 1122:1123 HA 1122:1123 HA 1122:1123 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 7 35 dep tagged 1126:1131 arg1 hemagglutinin 1107:1119 hemagglutinin 1107:1119 hemagglutinin (HA) 1107:1124 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 6 36 theme full 951:954 arg1 activity 966:973 full transport activity 951:973 full transport activity 951:973 Single mutants retained full transport activity; the double mutant retained a majority of activity. 18405659 8 37 theme Wild-type 1259:1267 arg1 HA 1294:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 0 38 theme transporter 118:128 arg1 function 74:81 function 74:81 function 74:81 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 38 theme transporter 118:128 arg1 mobility 61:68 electrophoretic mobility 45:68 electrophoretic mobility 45:68 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 3 39 theme protein 561:567 arg1 aggregation 542:552 aggregation 542:552 aggregation of the protein 542:567 Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. 18405659 0 40 dep mobility 61:68 arg1 the 41:43 the 41:43 the 41:43 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 5 41 theme asparagine 754:763 arg1 residues 765:772 asparagine residues 754:772 asparagine residues of two canonical glycosylation sites 754:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 5 41 theme asparagine 754:763 arg1 sites 805:809 two canonical glycosylation sites 777:809 two canonical glycosylation sites 777:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 7 42 theme COOH 1156:1159 arg1 terminus 1161:1168 COOH terminus 1156:1168 COOH terminus 1156:1168 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 3 43 theme Western 417:423 arg1 analysis 430:437 Western blot analysis 417:437 Western blot analysis 417:437 Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. 18405659 6 44 theme activity 1017:1024 arg1 majority 1005:1012 a majority 1003:1012 a majority of activity 1003:1024 Single mutants retained full transport activity; the double mutant retained a majority of activity. 18405659 7 45 theme protein 1250:1256 arg1 degradation 1216:1226 degradation 1216:1226 degradation of the deglycosylated protein 1216:1256 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 5 46 theme sites 805:809 arg1 residues 765:772 asparagine residues 754:772 asparagine residues of two canonical glycosylation sites 754:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 5 46 theme sites 805:809 arg1 sites 805:809 two canonical glycosylation sites 777:809 two canonical glycosylation sites 777:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 5 47 theme residues 765:772 arg1 Substitution 738:749 Substitution 738:749 Substitution of asparagine residues of two canonical glycosylation sites to glutamine 738:822 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 7 48 theme molecular 1050:1058 arg1 size 1060:1063 molecular size 1050:1063 molecular size 1050:1063 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 5 49 theme glycosylation 791:803 arg1 sites 805:809 two canonical glycosylation sites 777:809 two canonical glycosylation sites 777:809 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 0 50 gly glycosylation 9:21 arg1 function 74:81 function 74:81 function 74:81 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 0 50 gly glycosylation 9:21 arg1 mobility 61:68 electrophoretic mobility 45:68 electrophoretic mobility 45:68 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 7 51 theme deglycosylated 1235:1248 arg1 protein 1250:1256 the deglycosylated protein 1231:1256 the deglycosylated protein 1231:1256 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 5 52 dep smaller 900:906 arg1 kDa 913:915 ~35 kDa 909:915 ~35 kDa 909:915 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 0 53 theme electrophoretic 45:59 arg1 mobility 61:68 electrophoretic mobility 45:68 electrophoretic mobility 45:68 N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). 18405659 4 54 with cells 669:673 arg1 F 663:663 peptidyl N-glycanase F 642:663 peptidyl N-glycanase F 642:663 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 6 55 theme Single 927:932 arg1 mutants 934:940 Single mutants 927:940 Single mutants 927:940 Single mutants retained full transport activity; the double mutant retained a majority of activity. 18405659 4 56 with Treatment 570:578 arg1 tunicamycin 680:690 tunicamycin 680:690 tunicamycin 680:690 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 8 57 theme HsPCFT 1287:1292 arg1 HA 1294:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 4 58 theme peptidyl 642:649 arg1 F 663:663 peptidyl N-glycanase F 642:663 peptidyl N-glycanase F 642:663 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 1 59 theme other 295:299 arg1 tissues 301:307 other tissues 295:307 other tissues 295:307 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 6 60 theme transport 956:964 arg1 activity 966:973 full transport activity 951:973 full transport activity 951:973 Single mutants retained full transport activity; the double mutant retained a majority of activity. 18405659 1 61 dep transporter 172:182 arg1 SLC46A1 193:199 SLC46A1 193:199 SLC46A1 193:199 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 1 61 dep transporter 172:182 arg1 HsPCFT 185:190 HsPCFT 185:190 HsPCFT 185:190 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 2 62 theme Western 313:319 arg1 blot 321:324 Western blot 313:324 Western blot 313:324 On Western blot, HsPCFT migrates as a broad band (~55 kDa), higher than predicted (~50 kDa) in cell lines. 18405659 4 63 theme kDa 725:727 arg1 species 729:735 a ~35 kDa species 719:735 a ~35 kDa species 719:735 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 3 64 theme degrees 523:529 arg1 C 531:531 50 degrees C 520:531 50 degrees C 520:531 Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. 18405659 4 65 theme HeLa 626:629 arg1 cells 631:635 HsPCFT-transfected HeLa cells 607:635 HsPCFT-transfected HeLa cells 607:635 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 1 66 theme intestinal 211:220 arg1 absorption 222:231 intestinal absorption 211:231 intestinal absorption of folates 211:242 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 5 67 theme smaller 900:906 arg1 protein 918:924 a smaller (~35 kDa) protein 898:924 a smaller (~35 kDa) protein 898:924 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 4 68 theme N-glycanase 651:661 arg1 F 663:663 peptidyl N-glycanase F 642:663 peptidyl N-glycanase F 642:663 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 8 69 theme plasma 1367:1372 arg1 membrane 1374:1381 the plasma membrane 1363:1381 the plasma membrane 1363:1381 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 5 70 theme sites 887:891 arg1 substitution 866:877 substitution 866:877 substitution of both sites 866:891 Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. 18405659 4 71 theme HsPCFT-transfected 607:624 arg1 cells 631:635 HsPCFT-transfected HeLa cells 607:635 HsPCFT-transfected HeLa cells 607:635 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 7 72 theme anti-HA 1189:1195 arg1 antibody 1197:1204 an anti-HA antibody 1186:1204 an anti-HA antibody excluding degradation of the deglycosylated protein 1186:1256 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 8 73 gly deglycosylated 1272:1285 arg1 HA 1294:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA 1259:1295 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 7 74 gly deglycosylated 1235:1248 arg1 protein 1250:1256 the deglycosylated protein 1231:1256 the deglycosylated protein 1231:1256 Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. 18405659 8 75 with consistent 1425:1434 arg1 location 1459:1466 the intracellular location 1441:1466 the intracellular location of these domains 1441:1483 Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. 18405659 4 76 from cells 631:635 arg1 Treatment 570:578 Treatment 570:578 Treatment 570:578 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 4 76 from cells 631:635 arg1 fractions 592:600 membrane fractions 583:600 membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells 583:673 Treatment of membrane fractions from HsPCFT-transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. 18405659 1 77 theme folates 236:242 arg1 transport 248:256 transport 248:256 transport of folates into the liver, brain and other tissues 248:307 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 18405659 1 77 theme folates 236:242 arg1 absorption 222:231 intestinal absorption 211:231 intestinal absorption of folates 211:242 The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. 15532026 1 0 theme mannose 287:293 arg1 M6P 308:310 M6P 308:310 M6P 308:310 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 1 0 theme mannose 287:293 arg1 6-phosphate 295:305 mannose 6-phosphate 287:305 mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes 287:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 9 1 theme overall 1677:1683 arg1 efficiency 1685:1694 the overall efficiency 1673:1694 the overall efficiency of M6P formation 1673:1711 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 9 2 from loss 1560:1563 arg1 subunit 1609:1615 the gamma subunit 1599:1615 the gamma subunit 1599:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 8 3 contain contain 1482:1488 arg1 cells 1476:1480 ML III cells 1469:1480 ML III cells 1469:1480 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 8 3 contain contain 1482:1488 arg2 dimer 1499:1503 a GNPTAG dimer 1490:1503 a GNPTAG dimer of reduced molecular mass 1490:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 5 4 theme protease 969:976 arg1 sorting/transport 933:949 the sorting/transport 929:949 the sorting/transport of this lysosomal protease 929:976 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 4 5 from gene 663:666 arg1 siblings 675:682 two siblings 671:682 two siblings with ML III 671:694 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 9 6 theme formation 1703:1711 arg1 localization 1646:1657 the intracellular localization 1628:1657 the intracellular localization of GNPTAG 1628:1667 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 9 6 theme formation 1703:1711 arg1 efficiency 1685:1694 the overall efficiency 1673:1694 the overall efficiency of M6P formation 1673:1711 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 6 7 theme ammonium 1004:1011 arg1 chloride 1013:1020 ammonium chloride 1004:1020 ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, 1004:1098 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 6 8 theme ML 1173:1174 arg1 fibroblasts 1180:1190 ML III fibroblasts 1173:1190 ML III fibroblasts 1173:1190 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 5 9 theme labeled 871:877 arg1 CtsD 892:895 CtsD 892:895 CtsD 892:895 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 5 9 theme labeled 871:877 arg1 D 889:889 metabolically labeled cathepsin D 857:889 metabolically labeled cathepsin D (CtsD) 857:896 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 1 10 theme initial 254:260 arg1 step 262:265 the initial step 250:265 the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes 250:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 4 11 theme elevated 704:711 arg1 activities 713:722 elevated activities 704:722 elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts 704:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 0 12 theme glycosylation 146:158 arg1 site 160:163 a used glycosylation site 139:163 a used glycosylation site 139:163 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 8 13 dep dimer 1455:1459 arg1 contain 1482:1488 contain 1482:1488 contain a GNPTAG dimer of reduced molecular mass 1482:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 9 14 from site 1591:1594 arg1 subunit 1609:1615 the gamma subunit 1599:1615 the gamma subunit 1599:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 8 15 theme reduced 1508:1514 arg1 mass 1526:1529 reduced molecular mass 1508:1529 reduced molecular mass 1508:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 0 16 from mutation 8:15 arg1 GNPTAG 80:85 GNPTAG 80:85 GNPTAG 80:85 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 0 16 from mutation 8:15 arg1 subunit 71:77 UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit 20:77 UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III 20:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 8 17 theme Western 1367:1373 arg1 analysis 1380:1387 Western blot analysis 1367:1387 Western blot analysis of extracts from control fibroblasts 1367:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 7 18 theme novel 1243:1247 arg1 c.347_349delACA 1270:1284 c.347_349delACA 1270:1284 c.347_349delACA (p.Asn116del) 1270:1298 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 7 18 theme novel 1243:1247 arg1 mutation 1260:1267 a novel homozygous mutation 1241:1267 a novel homozygous mutation 1241:1267 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 1 19 theme lysosomal 370:378 arg1 enzymes 380:386 soluble lysosomal enzymes 362:386 soluble lysosomal enzymes 362:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 5 20 from fibroblasts 903:913 arg1 Immunoprecipitation 834:852 Immunoprecipitation 834:852 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts 834:913 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 5 21 theme lysosomal 959:967 arg1 protease 969:976 this lysosomal protease 954:976 this lysosomal protease 954:976 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 0 22 from subunit 71:77 arg1 siblings 95:102 two siblings 91:102 two siblings with mucolipidosis type III 91:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 8 23 theme extracts 1392:1399 arg1 analysis 1380:1387 Western blot analysis 1367:1387 Western blot analysis of extracts from control fibroblasts 1367:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 0 24 theme mucolipidosis 109:121 arg1 type 123:126 mucolipidosis type III 109:130 mucolipidosis type III 109:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 9 25 theme glycosylation 1577:1589 arg1 site 1591:1594 the used glycosylation site 1568:1594 the used glycosylation site in the gamma subunit 1568:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 4 26 theme lysosomal 735:743 arg1 enzymes 745:751 several lysosomal enzymes 727:751 several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts 727:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 2 27 theme multisubunit 417:428 arg1 enzyme 430:435 a multisubunit enzyme 415:435 a multisubunit enzyme composed of three subunits (alpha2beta2gamma2) that are products of two genes 415:513 The phosphotransferase is a multisubunit enzyme composed of three subunits (alpha2beta2gamma2) that are products of two genes. 15532026 2 27 theme multisubunit 417:428 arg1 phosphotransferase 393:410 The phosphotransferase 389:410 The phosphotransferase 389:410 The phosphotransferase is a multisubunit enzyme composed of three subunits (alpha2beta2gamma2) that are products of two genes. 15532026 7 28 gly glycosylation 1347:1359 arg2 site 1361:1364 a potential N-linked glycosylation site 1326:1364 a potential N-linked glycosylation site 1326:1364 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 6 29 dep 20 1121:1122 arg1 to 1118:1119 to 1118:1119 to 1118:1119 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 1 30 theme residues 313:320 arg1 formation 274:282 the formation 270:282 the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes 270:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 9 31 theme gamma 1603:1607 arg1 subunit 1609:1615 the gamma subunit 1599:1615 the gamma subunit 1599:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 9 32 from subunit 1609:1615 arg1 loss 1560:1563 the loss 1556:1563 the loss of the used glycosylation site in the gamma subunit 1556:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 7 33 theme N-linked 1338:1345 arg1 site 1361:1364 a potential N-linked glycosylation site 1326:1364 a potential N-linked glycosylation site 1326:1364 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 2 34 theme genes 509:513 arg1 subunits 455:462 three subunits 449:462 three subunits (alpha2beta2gamma2) that are products of two genes 449:513 The phosphotransferase is a multisubunit enzyme composed of three subunits (alpha2beta2gamma2) that are products of two genes. 15532026 2 34 theme genes 509:513 arg1 products 493:500 products 493:500 products of two genes 493:513 The phosphotransferase is a multisubunit enzyme composed of three subunits (alpha2beta2gamma2) that are products of two genes. 15532026 0 35 theme UDP-N-acetylglucosamine-1-phosphotransferase 20:63 arg1 GNPTAG 80:85 GNPTAG 80:85 GNPTAG 80:85 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 0 35 theme UDP-N-acetylglucosamine-1-phosphotransferase 20:63 arg1 subunit 71:77 UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit 20:77 UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III 20:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 6 36 theme receptor 1078:1085 arg1 interaction 1087:1097 the CtsD-M6P receptor interaction 1065:1097 the CtsD-M6P receptor interaction 1065:1097 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 8 37 theme glycosylated 1442:1453 arg1 dimer 1455:1459 a 97 kDa glycosylated dimer 1433:1459 a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass 1433:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 1 38 theme efficient 339:347 arg1 transport 349:357 the efficient transport 335:357 the efficient transport of soluble lysosomal enzymes 335:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 4 39 from activities 713:722 arg1 fibroblasts 765:775 cultured fibroblasts serum and diminished activities 756:807 cultured fibroblasts serum and diminished activities in cultured fibroblasts 756:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 6 40 theme synthesized 1138:1148 arg1 CtsD 1150:1153 the newly synthesized CtsD 1128:1153 the newly synthesized CtsD 1128:1153 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 4 41 theme cultured 812:819 arg1 fibroblasts 821:831 cultured fibroblasts 812:831 cultured fibroblasts 812:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 0 42 gly glycosylation 146:158 arg2 site 160:163 a used glycosylation site 139:163 a used glycosylation site 139:163 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 9 43 theme GNPTAG 1662:1667 arg1 localization 1646:1657 the intracellular localization 1628:1657 the intracellular localization of GNPTAG 1628:1667 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 9 43 theme GNPTAG 1662:1667 arg1 efficiency 1685:1694 the overall efficiency 1673:1694 the overall efficiency of M6P formation 1673:1711 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 8 44 from fibroblasts 1414:1424 arg1 extracts 1392:1399 extracts 1392:1399 extracts from control fibroblasts 1392:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 8 44 from fibroblasts 1414:1424 arg1 analysis 1380:1387 Western blot analysis 1367:1387 Western blot analysis of extracts from control fibroblasts 1367:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 8 45 theme mass 1526:1529 arg1 dimer 1499:1503 a GNPTAG dimer 1490:1503 a GNPTAG dimer of reduced molecular mass 1490:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 1 46 theme 6-phosphate 295:305 arg1 residues 313:320 mannose 6-phosphate (M6P) residues 287:320 mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes 287:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 7 47 link N-linked 1338:1345 arg1 site 1361:1364 a potential N-linked glycosylation site 1326:1364 a potential N-linked glycosylation site 1326:1364 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 6 48 theme M6P-dependent 1198:1210 arg1 manner 1212:1217 an M6P-dependent manner 1195:1217 an M6P-dependent manner 1195:1217 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 9 49 theme M6P 1699:1701 arg1 formation 1703:1711 M6P formation 1699:1711 M6P formation 1699:1711 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 4 50 theme GNPTAG 656:661 arg1 gene 663:666 the GNPTAG gene 652:666 the GNPTAG gene in two siblings with ML III 652:694 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 4 51 with siblings 675:682 arg1 ML 689:690 ML III 689:694 ML III 689:694 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 6 52 theme chloride 1013:1020 arg1 Addition 992:999 Addition 992:999 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, 992:1098 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 0 53 with siblings 95:102 arg1 type 123:126 mucolipidosis type III 109:130 mucolipidosis type III 109:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 0 54 theme used 141:144 arg1 site 160:163 a used glycosylation site 139:163 a used glycosylation site 139:163 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 5 55 theme cathepsin 879:887 arg1 CtsD 892:895 CtsD 892:895 CtsD 892:895 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 5 55 theme cathepsin 879:887 arg1 D 889:889 metabolically labeled cathepsin D 857:889 metabolically labeled cathepsin D (CtsD) 857:896 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 8 56 theme GNPTAG 1492:1497 arg1 dimer 1499:1503 a GNPTAG dimer 1490:1503 a GNPTAG dimer of reduced molecular mass 1490:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 3 57 theme mucolipidosis 602:614 arg1 type 616:619 mucolipidosis type III 602:623 mucolipidosis type III (ML III) 602:632 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 3 57 theme mucolipidosis 602:614 arg1 ML 626:627 ML III 626:631 ML III 626:631 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 3 58 from defective 575:583 arg1 patients 588:595 patients 588:595 patients with mucolipidosis type III (ML III) 588:632 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 1 59 from step 262:265 arg1 formation 274:282 the formation 270:282 the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes 270:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 6 60 theme CtsD-M6P 1069:1076 arg1 interaction 1087:1097 the CtsD-M6P receptor interaction 1065:1097 the CtsD-M6P receptor interaction 1065:1097 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 8 61 theme molecular 1516:1524 arg1 mass 1526:1529 reduced molecular mass 1508:1529 reduced molecular mass 1508:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 3 62 from patients 588:595 arg1 gene 520:523 The gene 516:523 The gene encoding the gamma-subunit (GNPTAG) 516:559 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 3 62 from patients 588:595 arg1 defective 575:583 defective 575:583 defective 575:583 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 7 63 theme homozygous 1249:1258 arg1 c.347_349delACA 1270:1284 c.347_349delACA 1270:1284 c.347_349delACA (p.Asn116del) 1270:1298 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 7 63 theme homozygous 1249:1258 arg1 mutation 1260:1267 a novel homozygous mutation 1241:1267 a novel homozygous mutation 1241:1267 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 8 64 dep glycosylated 1442:1453 arg1 kDa 1438:1440 97 kDa 1435:1440 97 kDa 1435:1440 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 1 65 theme soluble 362:368 arg1 enzymes 380:386 soluble lysosomal enzymes 362:386 soluble lysosomal enzymes 362:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 4 66 dep fibroblasts 765:775 arg1 serum 777:781 serum 777:781 serum 777:781 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 4 66 dep fibroblasts 765:775 arg1 activities 798:807 diminished activities 787:807 diminished activities 787:807 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 7 67 theme direct 1223:1228 arg1 sequencing 1230:1239 direct sequencing 1223:1239 direct sequencing 1223:1239 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 1 68 theme enzymes 380:386 arg1 transport 349:357 the efficient transport 335:357 the efficient transport of soluble lysosomal enzymes 335:386 The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. 15532026 4 69 from fibroblasts 765:775 arg1 activities 713:722 elevated activities 704:722 elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts 704:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 4 69 from fibroblasts 765:775 arg1 fibroblasts 821:831 cultured fibroblasts 812:831 cultured fibroblasts 812:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 6 70 theme pH-dependent 1033:1044 arg1 processes 1046:1054 pH-dependent processes 1033:1054 pH-dependent processes 1033:1054 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 6 70 theme pH-dependent 1033:1044 arg1 interaction 1087:1097 the CtsD-M6P receptor interaction 1065:1097 the CtsD-M6P receptor interaction 1065:1097 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 8 71 theme blot 1375:1378 arg1 analysis 1380:1387 Western blot analysis 1367:1387 Western blot analysis of extracts from control fibroblasts 1367:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 4 72 theme cultured 756:763 arg1 fibroblasts 765:775 cultured fibroblasts serum and diminished activities 756:807 cultured fibroblasts serum and diminished activities in cultured fibroblasts 756:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 9 73 theme used 1572:1575 arg1 site 1591:1594 the used glycosylation site 1568:1594 the used glycosylation site in the gamma subunit 1568:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 0 74 theme novel 2:6 arg1 mutation 8:15 A novel mutation 0:15 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III 0:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 4 75 from enzymes 745:751 arg1 fibroblasts 765:775 cultured fibroblasts serum and diminished activities 756:807 cultured fibroblasts serum and diminished activities in cultured fibroblasts 756:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 9 76 theme site 1591:1594 arg1 loss 1560:1563 the loss 1556:1563 the loss of the used glycosylation site in the gamma subunit 1556:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 7 77 theme glycosylation 1347:1359 arg1 site 1361:1364 a potential N-linked glycosylation site 1326:1364 a potential N-linked glycosylation site 1326:1364 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 4 78 theme several 727:733 arg1 enzymes 745:751 several lysosomal enzymes 727:751 several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts 727:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 3 79 with patients 588:595 arg1 type 616:619 mucolipidosis type III 602:623 mucolipidosis type III (ML III) 602:632 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 3 79 with patients 588:595 arg1 ML 626:627 ML III 626:631 ML III 626:631 The gene encoding the gamma-subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). 15532026 7 80 theme potential 1328:1336 arg1 site 1361:1364 a potential N-linked glycosylation site 1326:1364 a potential N-linked glycosylation site 1326:1364 By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. 15532026 8 81 theme control 1406:1412 arg1 fibroblasts 1414:1424 control fibroblasts 1406:1424 control fibroblasts 1406:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 9 82 gly glycosylation 1577:1589 arg2 site 1591:1594 the used glycosylation site 1568:1594 the used glycosylation site in the gamma subunit 1568:1615 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 4 83 theme enzymes 745:751 arg1 activities 713:722 elevated activities 704:722 elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts 704:831 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 0 84 theme gamma 65:69 arg1 GNPTAG 80:85 GNPTAG 80:85 GNPTAG 80:85 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 0 84 theme gamma 65:69 arg1 subunit 71:77 UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit 20:77 UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III 20:130 A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. 15532026 9 85 theme intracellular 1632:1644 arg1 localization 1646:1657 the intracellular localization 1628:1657 the intracellular localization of GNPTAG 1628:1667 These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. 15532026 6 86 theme CtsD 1150:1153 arg1 CtsD 1150:1153 the newly synthesized CtsD 1128:1153 the newly synthesized CtsD 1128:1153 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 6 86 theme CtsD 1150:1153 arg1 % 1123:1123 15 to 20% 1115:1123 15 to 20% of the newly synthesized CtsD 1115:1153 Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. 15532026 4 87 theme diminished 787:796 arg1 activities 798:807 diminished activities 787:807 diminished activities 787:807 We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. 15532026 8 88 from analysis 1380:1387 arg1 fibroblasts 1414:1424 control fibroblasts 1406:1424 control fibroblasts 1406:1424 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 5 89 theme D 889:889 arg1 Immunoprecipitation 834:852 Immunoprecipitation 834:852 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts 834:913 Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. 15532026 8 90 theme ML 1469:1470 arg1 cells 1476:1480 ML III cells 1469:1480 ML III cells 1469:1480 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 15532026 8 91 gly glycosylated 1442:1453 arg1 dimer 1455:1459 a 97 kDa glycosylated dimer 1433:1459 a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass 1433:1529 Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. 17711303 12 0 theme receptor 1969:1976 arg1 regulation 1978:1987 agonist-induced receptor regulation 1953:1987 agonist-induced receptor regulation 1953:1987 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 12 1 theme receptor 1925:1932 arg1 protein 1934:1940 the receptor protein 1921:1940 the receptor protein 1921:1940 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 4 2 theme Peptide-N-glycosidase 588:608 arg1 digestion 612:620 Peptide-N-glycosidase F digestion 588:620 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin 588:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 2 3 theme 55-kDa 284:289 arg1 FLAG-hKOR 238:246 FLAG-hKOR 238:246 FLAG-hKOR 238:246 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 2 3 theme 55-kDa 284:289 arg1 band 291:294 a broad and diffuse 55-kDa band 264:294 band 291:294 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 8 4 contain had 1199:1201 arg2 level 1222:1226 a lower expression level 1203:1226 a lower expression level than the wild type 1203:1245 When transiently expressed, hKOR-N25/39Q had a lower expression level than the wild type. 17711303 8 4 contain had 1199:1201 arg1 hKOR-N25/39Q 1186:1197 hKOR-N25/39Q 1186:1197 hKOR-N25/39Q 1186:1197 When transiently expressed, hKOR-N25/39Q had a lower expression level than the wild type. 17711303 10 5 theme rate 1484:1487 arg1 constant 1489:1496 the maturation rate constant 1469:1496 the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q 1469:1536 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 5 theme rate 1484:1487 arg1 times 1544:1548 6 times 1542:1548 6 times that of the mature form of the wild type 1542:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 5 theme rate 1484:1487 arg1 ka 1499:1500 ka 1499:1500 ka 1499:1500 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 3 6 theme 45-kDa 517:522 arg1 band 524:527 the 45-kDa band 513:527 the 45-kDa band 513:527 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 3 6 theme 45-kDa 517:522 arg1 N-glycosylated 532:545 N-glycosylated 532:545 N-glycosylated 532:545 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 9 7 theme intermediate 1375:1386 arg1 forms 1399:1403 the intermediate and mature forms 1371:1403 the intermediate and mature forms 1371:1403 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 12 8 theme biosynthesis 1897:1908 arg1 pathway 1910:1916 the biosynthesis pathway 1893:1916 the biosynthesis pathway of the receptor protein 1893:1940 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 7 9 theme mutant 1060:1065 arg1 hKOR-N25/39Q 1067:1078 The double mutant hKOR-N25/39Q 1049:1078 The double mutant hKOR-N25/39Q 1049:1078 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 7 9 theme mutant 1060:1065 arg1 38-kDa 1125:1130 38-kDa 1125:1130 38-kDa 1125:1130 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 7 9 theme mutant 1060:1065 arg1 43-kDa 1098:1103 43-kDa 1098:1103 43-kDa 1098:1103 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 9 10 theme mature 1392:1397 arg1 forms 1399:1403 the intermediate and mature forms 1371:1403 the intermediate and mature forms 1371:1403 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 6 11 theme N-terminal 943:952 arg1 domain 954:959 the N-terminal domain 939:959 the N-terminal domain 939:959 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain reduced the Mr by approximately 5 kDa, indicating that both residues were glycosylated. 17711303 8 12 theme expression 1211:1220 arg1 level 1222:1226 a lower expression level 1203:1226 a lower expression level than the wild type 1203:1245 When transiently expressed, hKOR-N25/39Q had a lower expression level than the wild type. 17711303 7 13 dep 43-kDa 1098:1103 arg1 band 1152:1155 band 1152:1155 band 1152:1155 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 0 14 theme biosynthesis 110:121 arg1 pathway 123:129 the biosynthesis pathway 106:129 the biosynthesis pathway 106:129 N-Glycosylation of the human kappa opioid receptor enhances its stability but slows its trafficking along the biosynthesis pathway. 17711303 10 15 theme hKOR-N25/39Q 1525:1536 arg1 form 1517:1520 the 43-kDa form 1506:1520 the 43-kDa form of hKOR-N25/39Q 1506:1536 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 11 16 theme ligand 1746:1751 arg1 affinity 1761:1768 ligand binding affinity 1746:1768 ligand binding affinity 1746:1768 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 5 17 theme Mr 806:807 arg1 bands 809:813 lower Mr bands 800:813 lower Mr bands 800:813 FLAG-hKOR was reduced to lower Mr bands by neuraminidase and O-glycosidase, indicating that the hKOR contains O-linked glycan. 17711303 1 18 theme functional 213:222 arg1 significance 224:235 its functional significance 209:235 its functional significance 209:235 We examined glycosylation of FLAG-hKOR expressed in CHO cells and determined its functional significance. 17711303 8 19 theme wild 1237:1240 arg1 type 1242:1245 the wild type 1233:1245 the wild type 1233:1245 When transiently expressed, hKOR-N25/39Q had a lower expression level than the wild type. 17711303 10 20 theme form 1517:1520 arg1 constant 1489:1496 the maturation rate constant 1469:1496 the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q 1469:1536 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 20 theme form 1517:1520 arg1 times 1544:1548 6 times 1542:1548 6 times that of the mature form of the wild type 1542:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 20 theme form 1517:1520 arg1 ka 1499:1500 ka 1499:1500 ka 1499:1500 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 2 21 dep diffuse 307:313 arg1 45-kDa 315:320 45-kDa 315:320 45-kDa 315:320 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 11 22 theme receptor 1649:1656 arg1 phosphorylation 1658:1672 increased agonist-induced receptor phosphorylation 1623:1672 increased agonist-induced receptor phosphorylation 1623:1672 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 4 23 theme kDa 719:721 arg1 species 698:704 two species 694:704 two species of 43 and 38 kDa 694:721 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 11 24 theme increased 1623:1631 arg1 phosphorylation 1658:1672 increased agonist-induced receptor phosphorylation 1623:1672 increased agonist-induced receptor phosphorylation 1623:1672 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 4 25 gly O-glycosylated 759:772 arg1 band 751:754 the 43-kDa band 740:754 the 43-kDa band 740:754 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 4 25 gly O-glycosylated 759:772 arg1 O-glycosylated 759:772 O-glycosylated 759:772 O-glycosylated 759:772 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 1 26 theme FLAG-hKOR 161:169 arg1 glycosylation 144:156 glycosylation 144:156 glycosylation of FLAG-hKOR expressed in CHO cells 144:192 We examined glycosylation of FLAG-hKOR expressed in CHO cells and determined its functional significance. 17711303 2 27 gly glycosylated 378:389 arg1 receptor 366:373 the receptor 362:373 the receptor 362:373 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 9 28 dep times 1426:1430 arg1 those 1432:1436 those 1432:1436 those 1432:1436 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 3 29 theme 45-kDa 422:427 arg1 band 429:432 the 45-kDa band 418:432 the 45-kDa band to approximately 38 kDa 418:456 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 5 30 theme lower 800:804 arg1 bands 809:813 lower Mr bands 800:813 lower Mr bands 800:813 FLAG-hKOR was reduced to lower Mr bands by neuraminidase and O-glycosidase, indicating that the hKOR contains O-linked glycan. 17711303 10 31 theme mature 1562:1567 arg1 form 1569:1572 the mature form 1558:1572 the mature form of the wild type 1558:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 0 32 theme kappa 29:33 arg1 receptor 42:49 the human kappa opioid receptor 19:49 the human kappa opioid receptor 19:49 N-Glycosylation of the human kappa opioid receptor enhances its stability but slows its trafficking along the biosynthesis pathway. 17711303 9 33 theme pulse-chase 1297:1307 arg1 experiments 1309:1319 pulse-chase experiments 1297:1319 pulse-chase experiments 1297:1319 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 10 34 from times 1544:1548 arg1 addition 1459:1466 addition 1459:1466 addition 1459:1466 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 12 35 theme protein 1934:1940 arg1 pathway 1910:1916 the biosynthesis pathway 1893:1916 the biosynthesis pathway of the receptor protein 1893:1940 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 3 36 theme Endoglycosidase 392:406 arg1 H 408:408 Endoglycosidase H 392:408 Endoglycosidase H 392:408 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 11 37 theme receptor-G 1773:1782 arg1 coupling 1792:1799 receptor-G protein coupling 1773:1799 receptor-G protein coupling 1773:1799 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 7 38 theme mature 1106:1111 arg1 form 1113:1116 mature form 1106:1116 mature form 1106:1116 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 7 38 theme mature 1106:1111 arg1 43-kDa 1098:1103 43-kDa 1098:1103 43-kDa 1098:1103 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 4 39 with incubation 645:654 arg1 tunicamycin 670:680 tunicamycin 670:680 tunicamycin 670:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 10 40 dep times 1544:1548 arg1 that 1550:1553 that 1550:1553 that 1550:1553 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 41 theme wild 1581:1584 arg1 type 1586:1589 the wild type 1577:1589 the wild type 1577:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 4 42 theme F 610:610 arg1 digestion 612:620 Peptide-N-glycosidase F digestion 588:620 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin 588:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 10 43 theme maturation 1473:1482 arg1 constant 1489:1496 the maturation rate constant 1469:1496 the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q 1469:1536 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 43 theme maturation 1473:1482 arg1 times 1544:1548 6 times 1542:1548 6 times that of the mature form of the wild type 1542:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 43 theme maturation 1473:1482 arg1 ka 1499:1500 ka 1499:1500 ka 1499:1500 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 4 44 with hKOR 637:640 arg1 tunicamycin 670:680 tunicamycin 670:680 tunicamycin 670:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 9 45 theme rate 1348:1351 arg1 ke 1364:1365 ke 1364:1365 ke 1364:1365 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 9 45 theme rate 1348:1351 arg1 times 1426:1430 approximately 3 times 1410:1430 approximately 3 times those of the wild type 1410:1453 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 9 45 theme rate 1348:1351 arg1 constants 1353:1361 the turnover rate constants 1335:1361 the turnover rate constants (ke) of the intermediate and mature forms 1335:1403 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 6 46 theme Asn25 914:918 arg1 Mutation 902:909 Mutation 902:909 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain 902:959 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain reduced the Mr by approximately 5 kDa, indicating that both residues were glycosylated. 17711303 12 47 theme agonist-induced 1953:1967 arg1 regulation 1978:1987 agonist-induced receptor regulation 1953:1987 agonist-induced receptor regulation 1953:1987 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 2 48 theme diffuse 276:282 arg1 FLAG-hKOR 238:246 FLAG-hKOR 238:246 FLAG-hKOR 238:246 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 2 48 theme diffuse 276:282 arg1 band 291:294 a broad and diffuse 55-kDa band 264:294 band 291:294 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 0 49 theme receptor 42:49 arg1 N-Glycosylation 0:14 N-Glycosylation 0:14 N-Glycosylation of the human kappa opioid receptor 0:49 N-Glycosylation of the human kappa opioid receptor enhances its stability but slows its trafficking along the biosynthesis pathway. 17711303 4 50 theme cells 659:663 arg1 incubation 645:654 incubation 645:654 incubation of cells with tunicamycin 645:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 4 50 theme cells 659:663 arg1 hKOR 637:640 solubilized hKOR 625:640 solubilized hKOR 625:640 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 10 51 theme 43-kDa 1510:1515 arg1 form 1517:1520 the 43-kDa form 1506:1520 the 43-kDa form of hKOR-N25/39Q 1506:1536 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 7 52 theme double 1053:1058 arg1 hKOR-N25/39Q 1067:1078 The double mutant hKOR-N25/39Q 1049:1078 The double mutant hKOR-N25/39Q 1049:1078 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 7 52 theme double 1053:1058 arg1 38-kDa 1125:1130 38-kDa 1125:1130 38-kDa 1125:1130 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 7 52 theme double 1053:1058 arg1 43-kDa 1098:1103 43-kDa 1098:1103 43-kDa 1098:1103 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 2 53 theme diffuse 307:313 arg1 FLAG-hKOR 238:246 FLAG-hKOR 238:246 FLAG-hKOR 238:246 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 2 53 theme diffuse 307:313 arg1 band 322:325 a less diffuse 45-kDa band 300:325 a less diffuse 45-kDa band by immunoblotting 300:343 FLAG-hKOR was resolved as a broad and diffuse 55-kDa band and a less diffuse 45-kDa band by immunoblotting, indicating that the receptor is glycosylated. 17711303 1 54 theme CHO 184:186 arg1 cells 188:192 CHO cells 184:192 CHO cells 184:192 We examined glycosylation of FLAG-hKOR expressed in CHO cells and determined its functional significance. 17711303 9 55 theme forms 1399:1403 arg1 ke 1364:1365 ke 1364:1365 ke 1364:1365 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 9 55 theme forms 1399:1403 arg1 times 1426:1430 approximately 3 times 1410:1430 approximately 3 times those of the wild type 1410:1453 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 9 55 theme forms 1399:1403 arg1 constants 1353:1361 the turnover rate constants 1335:1361 the turnover rate constants (ke) of the intermediate and mature forms 1335:1403 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 8 56 theme lower 1205:1209 arg1 level 1222:1226 a lower expression level 1203:1226 a lower expression level than the wild type 1203:1245 When transiently expressed, hKOR-N25/39Q had a lower expression level than the wild type. 17711303 3 57 theme 55-kDa 481:486 arg1 band 488:491 the 55-kDa band 477:491 the 55-kDa band 477:491 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 4 58 theme incubation 645:654 arg1 digestion 612:620 Peptide-N-glycosidase F digestion 588:620 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin 588:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 11 59 theme binding 1753:1759 arg1 affinity 1761:1768 ligand binding affinity 1746:1768 ligand binding affinity 1746:1768 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 1 60 gly glycosylation 144:156 arg1 FLAG-hKOR 161:169 FLAG-hKOR 161:169 FLAG-hKOR 161:169 We examined glycosylation of FLAG-hKOR expressed in CHO cells and determined its functional significance. 17711303 3 61 theme hybrid-type 568:578 arg1 glycan 580:585 hybrid-type glycan 568:585 hybrid-type glycan 568:585 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 6 62 theme Asn39 923:927 arg1 Mutation 902:909 Mutation 902:909 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain 902:959 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain reduced the Mr by approximately 5 kDa, indicating that both residues were glycosylated. 17711303 3 63 gly N-glycosylated 532:545 arg1 band 524:527 the 45-kDa band 513:527 the 45-kDa band 513:527 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 3 63 gly N-glycosylated 532:545 arg1 N-glycosylated 532:545 N-glycosylated 532:545 N-glycosylated 532:545 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 9 64 theme wild 1445:1448 arg1 type 1450:1453 the wild type 1441:1453 the wild type 1441:1453 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 5 65 contain contains 876:883 arg1 hKOR 871:874 the hKOR 867:874 the hKOR 867:874 FLAG-hKOR was reduced to lower Mr bands by neuraminidase and O-glycosidase, indicating that the hKOR contains O-linked glycan. 17711303 5 65 contain contains 876:883 arg2 glycan 894:899 O-linked glycan 885:899 O-linked glycan 885:899 FLAG-hKOR was reduced to lower Mr bands by neuraminidase and O-glycosidase, indicating that the hKOR contains O-linked glycan. 17711303 12 66 gly N-glycosylation 1808:1822 arg1 hKOR 1831:1834 the hKOR 1827:1834 the hKOR 1827:1834 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 11 67 theme agonist-induced 1633:1647 arg1 phosphorylation 1658:1672 increased agonist-induced receptor phosphorylation 1623:1672 increased agonist-induced receptor phosphorylation 1623:1672 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 12 68 theme important 1842:1850 arg1 roles 1852:1856 important roles 1842:1856 important roles 1842:1856 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 5 69 theme O-linked 885:892 arg1 glycan 894:899 O-linked glycan 885:899 O-linked glycan 885:899 FLAG-hKOR was reduced to lower Mr bands by neuraminidase and O-glycosidase, indicating that the hKOR contains O-linked glycan. 17711303 4 70 theme 43-kDa 744:749 arg1 band 751:754 the 43-kDa band 740:754 the 43-kDa band 740:754 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 4 70 theme 43-kDa 744:749 arg1 O-glycosylated 759:772 O-glycosylated 759:772 O-glycosylated 759:772 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 7 71 theme intermediate 1133:1144 arg1 38-kDa 1125:1130 38-kDa 1125:1130 38-kDa 1125:1130 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 7 71 theme intermediate 1133:1144 arg1 form 1146:1149 intermediate form 1133:1149 intermediate form 1133:1149 The double mutant hKOR-N25/39Q was resolved as a 43-kDa (mature form) and a 38-kDa (intermediate form) band. 17711303 9 72 theme CHO 1251:1253 arg1 cells 1255:1259 CHO cells 1251:1259 CHO cells stably expressing the hKOR-N25/39Q 1251:1294 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 4 73 theme hKOR 637:640 arg1 digestion 612:620 Peptide-N-glycosidase F digestion 588:620 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin 588:680 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 17711303 3 74 with N-glycosylated 532:545 arg1 high-mannose 552:563 high-mannose 552:563 high-mannose 552:563 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 3 74 with N-glycosylated 532:545 arg1 glycan 580:585 hybrid-type glycan 568:585 hybrid-type glycan 568:585 Endoglycosidase H cleaved the 45-kDa band to approximately 38 kDa but did not change the 55-kDa band, demonstrating that the 45-kDa band is N-glycosylated with high-mannose or hybrid-type glycan. 17711303 12 75 theme hKOR 1831:1834 arg1 N-glycosylation 1808:1822 N-glycosylation 1808:1822 N-glycosylation of the hKOR 1808:1834 Thus, N-glycosylation of the hKOR plays important roles in stability and trafficking along the biosynthesis pathway of the receptor protein as well as agonist-induced receptor regulation. 17711303 6 76 from Mutation 902:909 arg1 domain 954:959 the N-terminal domain 939:959 the N-terminal domain 939:959 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain reduced the Mr by approximately 5 kDa, indicating that both residues were glycosylated. 17711303 11 77 theme hKOR-N25/39Q 1596:1607 arg1 mutant 1609:1614 The hKOR-N25/39Q mutant 1592:1614 The hKOR-N25/39Q mutant 1592:1614 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 0 78 theme human 23:27 arg1 receptor 42:49 the human kappa opioid receptor 19:49 the human kappa opioid receptor 19:49 N-Glycosylation of the human kappa opioid receptor enhances its stability but slows its trafficking along the biosynthesis pathway. 17711303 5 79 link O-linked 885:892 arg1 glycan 894:899 O-linked glycan 885:899 O-linked glycan 885:899 FLAG-hKOR was reduced to lower Mr bands by neuraminidase and O-glycosidase, indicating that the hKOR contains O-linked glycan. 17711303 10 80 theme form 1569:1572 arg1 constant 1489:1496 the maturation rate constant 1469:1496 the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q 1469:1536 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 10 80 theme form 1569:1572 arg1 times 1544:1548 6 times 1542:1548 6 times that of the mature form of the wild type 1542:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 0 81 theme opioid 35:40 arg1 receptor 42:49 the human kappa opioid receptor 19:49 the human kappa opioid receptor 19:49 N-Glycosylation of the human kappa opioid receptor enhances its stability but slows its trafficking along the biosynthesis pathway. 17711303 0 82 gly N-Glycosylation 0:14 arg1 receptor 42:49 the human kappa opioid receptor 19:49 the human kappa opioid receptor 19:49 N-Glycosylation of the human kappa opioid receptor enhances its stability but slows its trafficking along the biosynthesis pathway. 17711303 10 83 theme type 1586:1589 arg1 form 1569:1572 the mature form 1558:1572 the mature form of the wild type 1558:1589 In addition, the maturation rate constant (ka) of the 43-kDa form of hKOR-N25/39Q was 6 times that of the mature form of the wild type. 17711303 11 84 theme protein 1784:1790 arg1 coupling 1792:1799 receptor-G protein coupling 1773:1799 receptor-G protein coupling 1773:1799 The hKOR-N25/39Q mutant showed increased agonist-induced receptor phosphorylation, desensitization, internalization, and downregulation, without changing ligand binding affinity or receptor-G protein coupling. 17711303 6 85 gly glycosylated 1035:1046 arg1 residues 1021:1028 both residues 1016:1028 both residues 1016:1028 Mutation of Asn25 or Asn39 to Gln in the N-terminal domain reduced the Mr by approximately 5 kDa, indicating that both residues were glycosylated. 17711303 9 86 theme turnover 1339:1346 arg1 ke 1364:1365 ke 1364:1365 ke 1364:1365 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 9 86 theme turnover 1339:1346 arg1 times 1426:1430 approximately 3 times 1410:1430 approximately 3 times those of the wild type 1410:1453 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 9 86 theme turnover 1339:1346 arg1 constants 1353:1361 the turnover rate constants 1335:1361 the turnover rate constants (ke) of the intermediate and mature forms 1335:1403 In CHO cells stably expressing the hKOR-N25/39Q, pulse-chase experiments revealed that the turnover rate constants (ke) of the intermediate and mature forms were approximately 3 times those of the wild type. 17711303 4 87 theme solubilized 625:635 arg1 hKOR 637:640 solubilized hKOR 625:640 solubilized hKOR 625:640 Peptide-N-glycosidase F digestion of solubilized hKOR or incubation of cells with tunicamycin resulted in two species of 43 and 38 kDa, suggesting that the 43-kDa band is O-glycosylated. 21138434 4 0 theme consensus 875:883 arg1 sequences 901:909 each consensus N-glycosylation sequences 870:909 each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 870:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 1 1 with channel 203:209 arg1 isoforms 236:243 potential molecular isoforms 216:243 potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 216:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 3 2 theme h5-HT3A 698:704 arg1 subunit 706:712 the h5-HT3A subunit 694:712 the h5-HT3A subunit 694:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 4 3 theme sequences 901:909 arg1 Disruption 856:865 Disruption 856:865 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 856:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 3 4 theme N-glycosylation 719:733 arg1 tunicamycin 745:755 the N-glycosylation inhibitor tunicamycin 715:755 the N-glycosylation inhibitor tunicamycin 715:755 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 4 5 theme h5-HT3A 1093:1099 arg1 subunit 1101:1107 the h5-HT3A subunit 1089:1107 the h5-HT3A subunit 1089:1107 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 3 6 theme protein 816:822 arg1 size 769:772 the size 765:772 the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa) 765:853 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 3 7 theme subunit 621:627 arg1 expression 595:604 transient expression 585:604 transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit 585:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 1 8 theme ligand-gated 186:197 arg1 receptor 161:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 1 8 theme ligand-gated 186:197 arg1 channel 203:209 a pentameric ligand-gated ion channel 173:209 a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 173:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 3 9 theme predominant 781:791 arg1 protein 816:822 predominant h5-HT3B-immunoreactive protein 781:822 the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa) 777:853 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 4 10 from subunit 926:932 arg1 Disruption 856:865 Disruption 856:865 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 856:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 5 11 dep sequences 1197:1205 arg1 or 1221:1222 or 1221:1222 or 1221:1222 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 3 12 theme kidney 648:653 arg1 cells 655:659 human embryonic kidney cells 632:659 human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit 632:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 3 12 theme kidney 648:653 arg1 cells 669:673 HEK293 cells 662:673 HEK293 cells 662:673 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 1 13 theme 5-hydroxytryptamine 129:147 arg1 receptor 161:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 1 13 theme 5-hydroxytryptamine 129:147 arg1 channel 203:209 a pentameric ligand-gated ion channel 173:209 a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 173:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 5 14 theme h5-HT3B 1271:1277 arg1 protein 1287:1293 the mutant h5-HT3B subunit protein 1260:1293 the mutant h5-HT3B subunit protein 1260:1293 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 3 15 theme ∼ 825:825 arg1 kDa 830:832 ∼ 55 kDa 825:832 ∼ 55 kDa reduced to ∼ 40 kDa 825:852 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 6 16 theme present 1364:1370 arg1 study 1372:1376 The present study 1360:1376 The present study 1360:1376 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 4 17 from sequences 901:909 arg1 subunit 926:932 the h5-HT3B subunit 914:932 the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 914:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 3 18 theme human 632:636 arg1 kidney 648:653 human embryonic kidney 632:653 human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit 632:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 2 19 theme 5-HT3B 494:499 arg1 subunit 501:507 the human (h) 5-HT3B subunit 480:507 the human (h) 5-HT3B subunit 480:507 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 0 20 theme subunit 75:81 arg1 subunit 75:81 the human 5-HT3B receptor subunit 49:81 the human 5-HT3B receptor subunit 49:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 0 20 theme subunit 75:81 arg1 residues 37:44 N-glycosylated residues 22:44 N-glycosylated residues of the human 5-HT3B receptor subunit 22:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 1 21 theme subunits 357:364 arg1 combinations 276:287 different subunit combinations 258:287 different subunit combinations 258:287 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 2 22 theme cell 532:535 arg1 expression 546:555 cell membrane expression 532:555 cell membrane expression 532:555 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 6 23 gly N-glycosylation 1402:1416 arg2 subunit 1439:1445 the h5-HT3B subunit 1427:1445 the h5-HT3B subunit 1427:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 6 23 gly N-glycosylation 1402:1416 arg1 subunit 1439:1445 the h5-HT3B subunit 1427:1445 the h5-HT3B subunit 1427:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 6 23 gly N-glycosylation 1402:1416 arg2 sites 1418:1422 utilised N-glycosylation sites 1393:1422 utilised N-glycosylation sites of the h5-HT3B subunit 1393:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 2 24 theme cell 419:422 arg1 trafficking 432:442 cell surface trafficking 419:442 cell surface trafficking 419:442 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 0 25 theme cell 99:102 arg1 expression 113:122 cell membrane expression 99:122 cell membrane expression 99:122 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 3 26 dep protein 816:822 arg1 kDa 830:832 ∼ 55 kDa 825:832 ∼ 55 kDa reduced to ∼ 40 kDa 825:852 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 0 27 theme receptor 66:73 arg1 subunit 75:81 the human 5-HT3B receptor subunit 49:81 the human 5-HT3B receptor subunit 49:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 2 28 theme subunit 403:409 arg1 N-glycosylation 373:387 N-glycosylation 373:387 N-glycosylation of the 5-HT3A subunit 373:409 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 1 29 theme different 258:266 arg1 combinations 276:287 different subunit combinations 258:287 different subunit combinations 258:287 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 5 30 theme protein 1287:1293 arg1 expression 1246:1255 the expression 1242:1255 the expression of the mutant h5-HT3B subunit protein in the cell membrane 1242:1314 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 2 31 theme h 491:491 arg1 subunit 501:507 the human (h) 5-HT3B subunit 480:507 the human (h) 5-HT3B subunit 480:507 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 6 32 theme utilised 1393:1400 arg1 subunit 1439:1445 the h5-HT3B subunit 1427:1445 the h5-HT3B subunit 1427:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 6 32 theme utilised 1393:1400 arg1 sites 1418:1422 utilised N-glycosylation sites 1393:1422 utilised N-glycosylation sites of the h5-HT3B subunit 1393:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 1 33 theme different 296:304 arg1 modifications 325:337 different post-translational modifications 296:337 different post-translational modifications 296:337 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 0 34 theme residues 37:44 arg1 identification 4:17 The identification 0:17 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 0:123 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 4 35 theme molecular 993:1001 arg1 kDa 1020:1022 2-4 kDa 1016:1022 2-4 kDa 1016:1022 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 4 35 theme molecular 993:1001 arg1 weight 1003:1008 a reduced molecular weight 983:1008 a reduced molecular weight (by ∼ 2-4 kDa) of each mutant 983:1038 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 0 36 theme 5-HT3B 59:64 arg1 subunit 75:81 the human 5-HT3B receptor subunit 49:81 the human 5-HT3B receptor subunit 49:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 0 37 gly N-glycosylated 22:35 arg1 subunit 75:81 the human 5-HT3B receptor subunit 49:81 the human 5-HT3B receptor subunit 49:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 0 37 gly N-glycosylated 22:35 arg1 residues 37:44 N-glycosylated residues 22:44 N-glycosylated residues of the human 5-HT3B receptor subunit 22:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 4 38 theme mutant 1033:1038 arg1 kDa 1020:1022 2-4 kDa 1016:1022 2-4 kDa 1016:1022 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 4 38 theme mutant 1033:1038 arg1 weight 1003:1008 a reduced molecular weight 983:1008 a reduced molecular weight (by ∼ 2-4 kDa) of each mutant 983:1038 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 6 39 theme subunit 1482:1488 arg1 expression 1490:1499 subunit expression 1482:1499 subunit expression in the cell membrane 1482:1520 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 1 40 theme ion 199:201 arg1 receptor 161:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 1 40 theme ion 199:201 arg1 channel 203:209 a pentameric ligand-gated ion channel 173:209 a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 173:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 2 41 theme human 484:488 arg1 subunit 501:507 the human (h) 5-HT3B subunit 480:507 the human (h) 5-HT3B subunit 480:507 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 5 42 theme Immunocytochemical 1110:1127 arg1 studies 1129:1135 Immunocytochemical studies 1110:1135 Immunocytochemical studies 1110:1135 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 1 43 theme molecular 226:234 arg1 isoforms 236:243 potential molecular isoforms 216:243 potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 216:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 6 44 theme subunit 1439:1445 arg1 subunit 1439:1445 the h5-HT3B subunit 1427:1445 the h5-HT3B subunit 1427:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 6 44 theme subunit 1439:1445 arg1 sites 1418:1422 utilised N-glycosylation sites 1393:1422 utilised N-glycosylation sites of the h5-HT3B subunit 1393:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 5 45 theme N-glycosylation 1181:1195 arg1 sequences 1197:1205 the N-glycosylation sequences 1177:1205 the N-glycosylation sequences (individually or combined) 1177:1232 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 4 46 theme N-glycosylation 885:899 arg1 sequences 901:909 each consensus N-glycosylation sequences 870:909 each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 870:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 5 47 from expression 1246:1255 arg1 membrane 1307:1314 the cell membrane 1298:1314 the cell membrane 1298:1314 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 3 48 theme inhibitor 735:743 arg1 tunicamycin 745:755 the N-glycosylation inhibitor tunicamycin 715:755 the N-glycosylation inhibitor tunicamycin 715:755 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 3 49 theme transient 585:593 arg1 expression 595:604 transient expression 585:604 transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit 585:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 4 50 theme HEK293 1058:1063 arg1 cells 1065:1069 HEK293 cells 1058:1069 HEK293 cells stably expressing the h5-HT3A subunit 1058:1107 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 6 51 theme cell 1508:1511 arg1 membrane 1513:1520 the cell membrane 1504:1520 the cell membrane 1504:1520 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 3 52 theme h5-HT3B 613:619 arg1 subunit 621:627 the h5-HT3B subunit 609:627 the h5-HT3B subunit 609:627 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 1 53 theme pentameric 175:184 arg1 receptor 161:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor 125:168 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 1 53 theme pentameric 175:184 arg1 channel 203:209 a pentameric ligand-gated ion channel 173:209 a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 173:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 2 54 theme membrane 537:544 arg1 expression 546:555 cell membrane expression 532:555 cell membrane expression 532:555 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 3 55 theme h5-HT3B-immunoreactive 793:814 arg1 protein 816:822 predominant h5-HT3B-immunoreactive protein 781:822 the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa) 777:853 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 3 56 theme embryonic 638:646 arg1 kidney 648:653 human embryonic kidney 632:653 human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit 632:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 6 57 theme receptor 1550:1557 arg1 function 1559:1566 5-HT(3) receptor function 1542:1566 5-HT(3) receptor function 1542:1566 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 4 58 theme h5-HT3B 918:924 arg1 subunit 926:932 the h5-HT3B subunit 914:932 the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 914:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 5 59 theme mutant 1264:1269 arg1 protein 1287:1293 the mutant h5-HT3B subunit protein 1260:1293 the mutant h5-HT3B subunit protein 1260:1293 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 4 60 dep subunit 926:932 arg1 N182S 964:968 N182S 964:968 N182S 964:968 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 4 60 dep subunit 926:932 arg1 N147S 954:958 N147S 954:958 N147S 954:958 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 4 60 dep subunit 926:932 arg1 N75S 941:944 N75S 941:944 N75S 941:944 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 2 61 theme subunit 501:507 arg1 N-glycosylation 461:475 N-glycosylation 461:475 N-glycosylation of the human (h) 5-HT3B subunit 461:507 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 1 62 theme individual 346:355 arg1 subunits 357:364 the individual subunits 342:364 the individual subunits 342:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 5 63 theme cell 1302:1305 arg1 membrane 1307:1314 the cell membrane 1298:1314 the cell membrane 1298:1314 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 3 64 theme ∼ 845:845 arg1 kDa 850:852 ∼ 40 kDa 845:852 ∼ 40 kDa 845:852 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 0 65 theme membrane 104:111 arg1 expression 113:122 cell membrane expression 99:122 cell membrane expression 99:122 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 4 66 from Disruption 856:865 arg1 subunit 926:932 the h5-HT3B subunit 914:932 the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) 914:969 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 6 67 theme h5-HT3B 1431:1437 arg1 subunit 1439:1445 the h5-HT3B subunit 1427:1445 the h5-HT3B subunit 1427:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 2 68 theme N-glycosylation 461:475 arg1 influence 517:525 the influence 513:525 the influence upon cell membrane expression 513:555 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 2 68 theme N-glycosylation 461:475 arg1 presence 449:456 the presence 445:456 the presence of N-glycosylation of the human (h) 5-HT3B subunit 445:507 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 5 69 theme subunit 1279:1285 arg1 protein 1287:1293 the mutant h5-HT3B subunit protein 1260:1293 the mutant h5-HT3B subunit protein 1260:1293 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 2 70 theme 5-HT3A 396:401 arg1 subunit 403:409 the 5-HT3A subunit 392:409 the 5-HT3A subunit 392:409 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 0 71 theme N-glycosylated 22:35 arg1 subunit 75:81 the human 5-HT3B receptor subunit 49:81 the human 5-HT3B receptor subunit 49:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 0 71 theme N-glycosylated 22:35 arg1 residues 37:44 N-glycosylated residues 22:44 N-glycosylated residues of the human 5-HT3B receptor subunit 22:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 1 72 theme subunit 268:274 arg1 combinations 276:287 different subunit combinations 258:287 different subunit combinations 258:287 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 6 73 dep identified 1382:1391 arg1 prerequisite 1525:1536 a prerequisite 1523:1536 a prerequisite for 5-HT(3) receptor function 1523:1566 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 3 74 theme HEK293 662:667 arg1 cells 655:659 human embryonic kidney cells 632:659 human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit 632:712 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 3 74 theme HEK293 662:667 arg1 cells 669:673 HEK293 cells 662:673 HEK293 cells 662:673 Following transient expression of the h5-HT3B subunit by human embryonic kidney cells (HEK293 cells) stably expressing the h5-HT3A subunit, the N-glycosylation inhibitor tunicamycin reduced the size of the predominant h5-HT3B-immunoreactive protein (∼ 55 kDa reduced to ∼ 40 kDa). 21138434 6 75 theme N-glycosylation 1402:1416 arg1 subunit 1439:1445 the h5-HT3B subunit 1427:1445 the h5-HT3B subunit 1427:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 6 75 theme N-glycosylation 1402:1416 arg1 sites 1418:1422 utilised N-glycosylation sites 1393:1422 utilised N-glycosylation sites of the h5-HT3B subunit 1393:1445 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 5 76 theme each 1169:1172 arg1 disruption 1155:1164 disruption 1155:1164 disruption of each of the N-glycosylation sequences (individually or combined) 1155:1232 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 2 77 gly N-glycosylation 373:387 arg1 subunit 403:409 the 5-HT3A subunit 392:409 the 5-HT3A subunit 392:409 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 2 78 theme surface 424:430 arg1 trafficking 432:442 cell surface trafficking 419:442 cell surface trafficking 419:442 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 0 79 theme human 53:57 arg1 subunit 75:81 the human 5-HT3B receptor subunit 49:81 the human 5-HT3B receptor subunit 49:81 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 1 80 theme post-translational 306:323 arg1 modifications 325:337 different post-translational modifications 296:337 different post-translational modifications 296:337 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 21138434 6 81 from expression 1490:1499 arg1 membrane 1513:1520 the cell membrane 1504:1520 the cell membrane 1504:1520 The present study has identified utilised N-glycosylation sites of the h5-HT3B subunit and demonstrated that they promote subunit expression in the cell membrane; a prerequisite for 5-HT(3) receptor function. 21138434 4 82 theme reduced 985:991 arg1 kDa 1020:1022 2-4 kDa 1016:1022 2-4 kDa 1016:1022 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 4 82 theme reduced 985:991 arg1 weight 1003:1008 a reduced molecular weight 983:1008 a reduced molecular weight (by ∼ 2-4 kDa) of each mutant 983:1038 Disruption of each consensus N-glycosylation sequences in the h5-HT3B subunit (N31S, N75S, N117S, N147S and N182S) resulted in a reduced molecular weight (by ∼ 2-4 kDa) of each mutant when expressed by HEK293 cells stably expressing the h5-HT3A subunit. 21138434 2 83 gly N-glycosylation 461:475 arg1 subunit 501:507 the human (h) 5-HT3B subunit 480:507 the human (h) 5-HT3B subunit 480:507 Since N-glycosylation of the 5-HT3A subunit impacts cell surface trafficking, the presence of N-glycosylation of the human (h) 5-HT3B subunit and the influence upon cell membrane expression was investigated. 21138434 0 84 dep identification 4:17 arg1 importance 84:93 importance 84:93 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 0:123 The identification of N-glycosylated residues of the human 5-HT3B receptor subunit: importance for cell membrane expression. 21138434 5 85 theme h5-HT3A 1343:1349 arg1 subunit 1351:1357 the h5-HT3A subunit 1339:1357 the h5-HT3A subunit 1339:1357 Immunocytochemical studies demonstrated that disruption of each of the N-glycosylation sequences (individually or combined) reduced the expression of the mutant h5-HT3B subunit protein in the cell membrane when co-expressed with the h5-HT3A subunit. 21138434 1 86 theme potential 216:224 arg1 isoforms 236:243 potential molecular isoforms 216:243 potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits 216:364 The 5-hydroxytryptamine 3 (5-HT(3)) receptor is a pentameric ligand-gated ion channel with potential molecular isoforms arising from different subunit combinations and/or different post-translational modifications of the individual subunits. 28402104 7 0 theme visual 1417:1422 arg1 GPCRs 1424:1428 monostable visual GPCRs 1406:1428 monostable visual GPCRs 1406:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 6 1 theme active 1022:1027 arg1 states 1029:1034 inactive and active states 1009:1034 inactive and active states of green cone opsin 1009:1054 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 5 2 gly glycosylated 956:967 arg1 sites 924:928 the predicted glycosylation sites 896:928 the predicted glycosylation sites at N32 and N34 896:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 6 3 theme extracellular 1110:1122 arg1 region 1135:1140 the extracellular N-terminal region 1106:1140 the extracellular N-terminal region 1106:1140 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 7 4 theme Pro-Pro 1359:1365 arg1 motif 1367:1371 a conserved Pro-Pro motif 1347:1371 a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs 1347:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 7 5 theme transmembrane 1258:1270 arg1 helix 1272:1276 transmembrane helix four (TM4) 1258:1287 transmembrane helix four (TM4) 1258:1287 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 6 6 theme site 1200:1203 arg1 accessibility 1159:1171 an increased accessibility 1146:1171 an increased accessibility of the chromophore binding site 1146:1203 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 6 6 theme site 1200:1203 arg1 accessibility 1089:1101 a reduced solvent accessibility 1071:1101 a reduced solvent accessibility of the extracellular N-terminal region 1071:1140 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 4 7 from opsin 631:635 arg1 extent 594:599 The extent 590:599 The extent of H/D exchange in green cone opsin 590:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 4 7 from opsin 631:635 arg1 greater 641:647 greater 641:647 greater 641:647 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 6 8 theme solvent 1081:1087 arg1 accessibility 1089:1101 a reduced solvent accessibility 1071:1101 a reduced solvent accessibility of the extracellular N-terminal region 1071:1140 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 6 9 theme chromophore 1180:1190 arg1 site 1200:1203 the chromophore binding site 1176:1203 the chromophore binding site 1176:1203 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 1 10 theme protein 195:201 arg1 responsible 302:312 responsible 302:312 responsible 302:312 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 1 10 theme protein 195:201 arg1 component 203:211 the protein component 191:211 the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal 191:368 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 7 11 theme alignments 1327:1336 arg1 analysis 1306:1313 an analysis 1303:1313 an analysis of sequence alignments 1303:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 4 12 theme exchange 608:615 arg1 extent 594:599 The extent 590:599 The extent of H/D exchange in green cone opsin 590:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 4 12 theme exchange 608:615 arg1 greater 641:647 greater 641:647 greater 641:647 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 4 13 from exchange 608:615 arg1 opsin 631:635 green cone opsin 620:635 green cone opsin 620:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 5 14 theme cone 804:807 arg1 opsin 809:813 green cone opsin 798:813 green cone opsin 798:813 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 6 15 theme deuterium 984:992 arg1 uptake 994:999 deuterium uptake 984:999 deuterium uptake 984:999 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 4 16 theme green 620:624 arg1 opsin 631:635 green cone opsin 620:635 green cone opsin 620:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 0 17 theme Monostable 128:137 arg1 Receptors 164:172 Monostable Visual G Protein-Coupled Receptors 128:172 Monostable Visual G Protein-Coupled Receptors 128:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 1 18 theme sensitive 222:230 arg1 receptors 250:258 light sensitive G protein-coupled receptors 216:258 light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye 216:291 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 1 18 theme sensitive 222:230 arg1 GPCRs 261:265 GPCRs 261:265 GPCRs 261:265 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 4 19 theme dark 674:677 arg1 states 692:697 the dark and bleached states 670:697 the dark and bleached states 670:697 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 1 20 from component 203:211 arg1 retina 275:280 the retina 271:280 the retina of the eye 271:291 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 4 21 theme bleached 683:690 arg1 states 692:697 the dark and bleached states 670:697 the dark and bleached states 670:697 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 8 22 from side 1522:1525 arg1 locus 1472:1476 the locus 1468:1476 the locus of chromophore release at the extracellular side of TM4 and TM5 1468:1540 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 0 23 theme Receptors 164:172 arg1 Loop 118:121 Extracellular Loop 2 104:123 Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors 104:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 2 24 theme green 483:487 arg1 opsin 494:498 green cone opsin 483:498 green cone opsin 483:498 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 0 25 from Motif 95:99 arg1 Loop 118:121 Extracellular Loop 2 104:123 Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors 104:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 6 26 theme region 1135:1140 arg1 accessibility 1159:1171 an increased accessibility 1146:1171 an increased accessibility of the chromophore binding site 1146:1203 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 6 26 theme region 1135:1140 arg1 accessibility 1089:1101 a reduced solvent accessibility 1071:1101 a reduced solvent accessibility of the extracellular N-terminal region 1071:1140 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 4 27 from extent 594:599 arg1 opsin 631:635 green cone opsin 620:635 green cone opsin 620:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 0 28 theme Conserved 77:85 arg1 Motif 95:99 a Conserved Pro-Pro Motif 75:99 a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors 75:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 5 29 theme inactive 847:854 arg1 dark 841:844 both dark (inactive) and bleached (active) states 836:884 dark 841:844 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 2 30 theme light 505:509 arg1 activation 511:520 light activation 505:520 light activation 505:520 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 4 31 theme structural 720:729 arg1 heterogeneity 731:743 a higher structural heterogeneity 711:743 a higher structural heterogeneity for green cone opsin 711:764 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 0 32 theme Extracellular 104:116 arg1 Loop 118:121 Extracellular Loop 2 104:123 Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors 104:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 1 33 theme biochemical 351:361 arg1 signal 363:368 a biochemical signal 349:368 a biochemical signal 349:368 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 5 34 theme predicted 900:908 arg1 sites 924:928 the predicted glycosylation sites 896:928 the predicted glycosylation sites at N32 and N34 896:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 0 35 theme Hydrogen/Deuterium 0:17 arg1 Spectrometry 33:44 Hydrogen/Deuterium Exchange Mass Spectrometry 0:44 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin 0:65 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 7 36 theme helix 1272:1276 arg1 side 1250:1253 the extracellular side 1232:1253 the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments 1232:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 2 37 theme H/D 405:407 arg1 exchange 410:417 hydrogen/deuterium (H/D) exchange 385:417 hydrogen/deuterium (H/D) exchange coupled with mass spectrometry 385:448 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 0 38 theme Mass 28:31 arg1 Spectrometry 33:44 Hydrogen/Deuterium Exchange Mass Spectrometry 0:44 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin 0:65 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 2 39 theme hydrogen/deuterium 385:402 arg1 exchange 410:417 hydrogen/deuterium (H/D) exchange 385:417 hydrogen/deuterium (H/D) exchange coupled with mass spectrometry 385:448 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 7 40 theme extracellular 1236:1248 arg1 side 1250:1253 the extracellular side 1232:1253 the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments 1232:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 8 41 theme release 1493:1499 arg1 locus 1472:1476 the locus 1468:1476 the locus of chromophore release at the extracellular side of TM4 and TM5 1468:1540 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 0 42 theme Green 55:59 arg1 Opsin 61:65 Human Green Opsin 49:65 Human Green Opsin 49:65 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 7 43 from side 1250:1253 arg1 exchange 1220:1227 Increased H/D exchange 1206:1227 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments 1206:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 6 44 theme opsin 1050:1054 arg1 states 1029:1034 inactive and active states 1009:1034 inactive and active states of green cone opsin 1009:1054 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 4 45 theme cone 755:758 arg1 opsin 760:764 green cone opsin 749:764 green cone opsin 749:764 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 7 46 theme GPCRs 1424:1428 arg1 loop 1390:1393 extracellular loop 2 1376:1395 extracellular loop 2 (EL2) of monostable visual GPCRs 1376:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 7 46 theme GPCRs 1424:1428 arg1 EL2 1398:1400 EL2 1398:1400 EL2 1398:1400 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 7 47 theme H/D 1216:1218 arg1 exchange 1220:1227 Increased H/D exchange 1206:1227 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments 1206:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 8 48 theme future 1571:1576 arg1 evaluation 1589:1598 future functional evaluation 1571:1598 future functional evaluation 1571:1598 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 1 49 theme receptors 250:258 arg1 responsible 302:312 responsible 302:312 responsible 302:312 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 1 49 theme receptors 250:258 arg1 component 203:211 the protein component 191:211 the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal 191:368 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 7 50 theme monostable 1406:1415 arg1 GPCRs 1424:1428 monostable visual GPCRs 1406:1428 monostable visual GPCRs 1406:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 8 51 theme TM4 1530:1532 arg1 side 1522:1525 the extracellular side 1504:1525 the extracellular side of TM4 and TM5 1504:1540 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 1 52 from retina 275:280 arg1 responsible 302:312 responsible 302:312 responsible 302:312 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 1 52 from retina 275:280 arg1 component 203:211 the protein component 191:211 the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal 191:368 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 8 53 theme TM5 1538:1540 arg1 side 1522:1525 the extracellular side 1504:1525 the extracellular side of TM4 and TM5 1504:1540 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 7 54 theme extracellular 1376:1388 arg1 loop 1390:1393 extracellular loop 2 1376:1395 extracellular loop 2 (EL2) of monostable visual GPCRs 1376:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 7 54 theme extracellular 1376:1388 arg1 EL2 1398:1400 EL2 1398:1400 EL2 1398:1400 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 5 55 from N32 933:935 arg1 sites 924:928 the predicted glycosylation sites 896:928 the predicted glycosylation sites at N32 and N34 896:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 4 56 theme cone 626:629 arg1 opsin 631:635 green cone opsin 620:635 green cone opsin 620:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 6 57 theme binding 1192:1198 arg1 site 1200:1203 the chromophore binding site 1176:1203 the chromophore binding site 1176:1203 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 6 58 theme reduced 1073:1079 arg1 accessibility 1089:1101 a reduced solvent accessibility 1071:1101 a reduced solvent accessibility of the extracellular N-terminal region 1071:1140 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 5 59 theme green 798:802 arg1 opsin 809:813 green cone opsin 798:813 green cone opsin 798:813 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 7 60 theme conserved 1349:1357 arg1 motif 1367:1371 a conserved Pro-Pro motif 1347:1371 a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs 1347:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 2 61 from changes 472:478 arg1 opsin 494:498 green cone opsin 483:498 green cone opsin 483:498 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 4 62 theme H/D 604:606 arg1 exchange 608:615 H/D exchange 604:615 H/D exchange in green cone opsin 604:635 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 0 63 theme Visual 139:144 arg1 Receptors 164:172 Monostable Visual G Protein-Coupled Receptors 128:172 Monostable Visual G Protein-Coupled Receptors 128:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 5 64 from N34 941:943 arg1 sites 924:928 the predicted glycosylation sites 896:928 the predicted glycosylation sites at N32 and N34 896:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 1 65 theme light 216:220 arg1 receptors 250:258 light sensitive G protein-coupled receptors 216:258 light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye 216:291 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 1 65 theme light 216:220 arg1 GPCRs 261:265 GPCRs 261:265 GPCRs 261:265 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 7 66 theme sequence 1318:1325 arg1 alignments 1327:1336 sequence alignments 1318:1336 sequence alignments 1318:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 2 67 used used 380:383 arg2 we 377:378 we 377:378 we 377:378 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 1 68 from receptors 250:258 arg1 retina 275:280 the retina 271:280 the retina of the eye 271:291 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 6 69 theme increased 1149:1157 arg1 accessibility 1159:1171 an increased accessibility 1146:1171 an increased accessibility of the chromophore binding site 1146:1203 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 5 70 theme bleached 861:868 arg1 states 879:884 both dark (inactive) and bleached (active) states 836:884 states 879:884 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 7 71 theme Increased 1206:1214 arg1 exchange 1220:1227 Increased H/D exchange 1206:1227 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments 1206:1336 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 6 72 theme N-terminal 1124:1133 arg1 region 1135:1140 the extracellular N-terminal region 1106:1140 the extracellular N-terminal region 1106:1140 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 0 73 theme Pro-Pro 87:93 arg1 Motif 95:99 a Conserved Pro-Pro Motif 75:99 a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors 75:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 7 74 from motif 1367:1371 arg1 loop 1390:1393 extracellular loop 2 1376:1395 extracellular loop 2 (EL2) of monostable visual GPCRs 1376:1428 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 7 74 from motif 1367:1371 arg1 EL2 1398:1400 EL2 1398:1400 EL2 1398:1400 Increased H/D exchange at the extracellular side of transmembrane helix four (TM4) combined with an analysis of sequence alignments revealed a conserved Pro-Pro motif in extracellular loop 2 (EL2) of monostable visual GPCRs. 28402104 2 75 theme conformational 457:470 arg1 changes 472:478 conformational changes 457:478 conformational changes in green cone opsin upon light activation 457:520 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 1 76 theme light 338:342 arg1 transduction 322:333 the transduction 318:333 the transduction of light into a biochemical signal 318:368 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 8 77 theme new 1450:1452 arg1 insights 1454:1461 new insights 1450:1461 new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 1450:1540 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 4 78 theme higher 713:718 arg1 heterogeneity 731:743 a higher structural heterogeneity 711:743 a higher structural heterogeneity for green cone opsin 711:764 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 6 79 theme green 1039:1043 arg1 opsin 1050:1054 green cone opsin 1039:1054 green cone opsin 1039:1054 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 6 80 theme inactive 1009:1016 arg1 states 1029:1034 inactive and active states 1009:1034 inactive and active states of green cone opsin 1009:1054 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 2 81 theme cone 489:492 arg1 opsin 494:498 green cone opsin 483:498 green cone opsin 483:498 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 0 82 theme Exchange 19:26 arg1 Spectrometry 33:44 Hydrogen/Deuterium Exchange Mass Spectrometry 0:44 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin 0:65 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 5 83 theme active 871:876 arg1 states 879:884 both dark (inactive) and bleached (active) states 836:884 states 879:884 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 6 84 theme uptake 994:999 arg1 Comparison 970:979 Comparison 970:979 Comparison of deuterium uptake between inactive and active states of green cone opsin 970:1054 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 8 85 theme chromophore 1481:1491 arg1 release 1493:1499 chromophore release 1481:1499 chromophore release 1481:1499 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 0 86 theme Protein-Coupled 148:162 arg1 Receptors 164:172 Monostable Visual G Protein-Coupled Receptors 128:172 Monostable Visual G Protein-Coupled Receptors 128:172 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 0 87 theme Human 49:53 arg1 Opsin 61:65 Human Green Opsin 49:65 Human Green Opsin 49:65 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 1 88 theme eye 289:291 arg1 retina 275:280 the retina 271:280 the retina of the eye 271:291 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 5 89 gly glycosylation 910:922 arg2 N34 941:943 N34 941:943 N34 941:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 5 89 gly glycosylation 910:922 arg2 N32 933:935 N32 933:935 N32 933:935 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 5 89 gly glycosylation 910:922 arg2 sites 924:928 the predicted glycosylation sites 896:928 the predicted glycosylation sites at N32 and N34 896:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 0 90 theme Opsin 61:65 arg1 Spectrometry 33:44 Hydrogen/Deuterium Exchange Mass Spectrometry 0:44 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin 0:65 Hydrogen/Deuterium Exchange Mass Spectrometry of Human Green Opsin Reveals a Conserved Pro-Pro Motif in Extracellular Loop 2 of Monostable Visual G Protein-Coupled Receptors. 28402104 2 91 theme mass 432:435 arg1 spectrometry 437:448 mass spectrometry 432:448 mass spectrometry 432:448 Here, we used hydrogen/deuterium (H/D) exchange coupled with mass spectrometry to map conformational changes in green cone opsin upon light activation. 28402104 8 92 theme extracellular 1508:1520 arg1 side 1522:1525 the extracellular side 1504:1525 the extracellular side of TM4 and TM5 1504:1540 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 5 93 theme glycosylation 910:922 arg1 sites 924:928 the predicted glycosylation sites 896:928 the predicted glycosylation sites at N32 and N34 896:943 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 28402104 6 94 theme cone 1045:1048 arg1 opsin 1050:1054 green cone opsin 1039:1054 green cone opsin 1039:1054 Comparison of deuterium uptake between inactive and active states of green cone opsin also disclosed a reduced solvent accessibility of the extracellular N-terminal region and an increased accessibility of the chromophore binding site. 28402104 4 95 theme green 749:753 arg1 opsin 760:764 green cone opsin 749:764 green cone opsin 749:764 The extent of H/D exchange in green cone opsin was greater than in rhodopsin in the dark and bleached states, suggesting a higher structural heterogeneity for green cone opsin. 28402104 8 96 theme functional 1578:1587 arg1 evaluation 1589:1598 future functional evaluation 1571:1598 future functional evaluation 1571:1598 These data present new insights into the locus of chromophore release at the extracellular side of TM4 and TM5 and provide a foundation for future functional evaluation. 28402104 1 97 theme protein-coupled 234:248 arg1 receptors 250:258 light sensitive G protein-coupled receptors 216:258 light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye 216:291 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 1 97 theme protein-coupled 234:248 arg1 GPCRs 261:265 GPCRs 261:265 GPCRs 261:265 Opsins comprise the protein component of light sensitive G protein-coupled receptors (GPCRs) in the retina of the eye that are responsible for the transduction of light into a biochemical signal. 28402104 5 98 theme Further 767:773 arg1 analysis 775:782 Further analysis 767:782 Further analysis 767:782 Further analysis revealed that green cone opsin exists as a dimer in both dark (inactive) and bleached (active) states, and that the predicted glycosylation sites at N32 and N34 are indeed glycosylated. 12022871 9 0 theme active 1342:1347 arg1 enzyme 1365:1370 an active nonglycosylated enzyme 1339:1370 an active nonglycosylated enzyme 1339:1370 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 10 1 theme retrieval 1431:1439 arg1 signal 1441:1446 the putative C-terminal endoplasmic reticulum retrieval signal 1385:1446 the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) 1385:1453 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 10 1 theme retrieval 1431:1439 arg1 HIEL 1449:1452 HIEL 1449:1452 HIEL 1449:1452 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 6 2 theme inactive 1062:1069 arg1 enzyme 1071:1076 a catalytically inactive enzyme 1046:1076 a catalytically inactive enzyme 1046:1076 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 3 3 from Expression 507:516 arg1 coli 546:549 Escherichia coli 534:549 Escherichia coli 534:549 Expression of human TGH in Escherichia coli yields a protein without enzymatic activity, which suggests that posttranslational processing is necessary for the catalytic activity. 12022871 8 4 gly glycosylated 1241:1252 arg2 TGH 1233:1235 Human TGH 1227:1235 Human TGH 1227:1235 Human TGH was glycosylated in the insect cells. 12022871 8 4 gly glycosylated 1241:1252 arg1 TGH 1233:1235 Human TGH 1227:1235 Human TGH 1227:1235 Human TGH was glycosylated in the insect cells. 12022871 8 4 gly glycosylated 1241:1252 arg1 cells 1268:1272 the insect cells 1257:1272 the insect cells 1257:1272 Human TGH was glycosylated in the insect cells. 12022871 2 5 theme enzyme 425:430 arg1 relationship 405:416 The structure-activity relationship 382:416 The structure-activity relationship of the enzyme 382:430 The structure-activity relationship of the enzyme was investigated by site-directed mutagenesis and heterologous expression. 12022871 11 6 theme alpha/beta 1556:1565 arg1 fold 1577:1580 a lipase alpha/beta hydrolase fold 1547:1580 a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285) 1547:1657 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 6 7 dep residues 1003:1010 arg1 E354A 1020:1024 E354A 1020:1024 E354A 1020:1024 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 6 7 dep residues 1003:1010 arg1 H468A 1031:1035 H468A 1031:1035 H468A 1031:1035 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 6 7 dep residues 1003:1010 arg1 S221A 1013:1017 S221A 1013:1017 S221A 1013:1017 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 6 7 dep residues 1003:1010 arg1 residues 1003:1010 the residues 999:1010 the residues (S221A, E354A, and H468A) 999:1036 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 0 8 theme catalytic 115:123 arg1 triad 125:129 the catalytic triad 111:129 the catalytic triad 111:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 6 9 theme residues 1003:1010 arg1 mutagenesis 984:994 Site-directed mutagenesis 970:994 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) 970:1036 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 10 10 theme protein 1497:1503 arg1 secretion 1473:1481 secretion 1473:1481 secretion of the mutant protein 1473:1503 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 4 11 theme sequence 788:795 arg1 processing 752:761 correct processing 744:761 correct processing of the N-terminal signal sequence 744:795 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 1 12 theme lipolysis/re-esterification 294:320 arg1 cycle 322:326 the lipolysis/re-esterification cycle 290:326 the lipolysis/re-esterification cycle 290:326 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 9 13 gly nonglycosylated 1349:1363 arg1 enzyme 1365:1370 an active nonglycosylated enzyme 1339:1370 an active nonglycosylated enzyme 1339:1370 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 10 14 theme mutant 1490:1495 arg1 protein 1497:1503 the mutant protein 1486:1503 the mutant protein 1486:1503 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 3 15 theme enzymatic 576:584 arg1 activity 586:593 enzymatic activity 576:593 enzymatic activity 576:593 Expression of human TGH in Escherichia coli yields a protein without enzymatic activity, which suggests that posttranslational processing is necessary for the catalytic activity. 12022871 4 16 theme active 825:830 arg1 enzyme 832:837 a catalytically active enzyme 809:837 a catalytically active enzyme 809:837 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 4 17 theme baculovirus-infected 700:719 arg1 cells 726:730 baculovirus-infected Sf-9 cells 700:730 baculovirus-infected Sf-9 cells 700:730 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 9 18 gly N-glycosylation 1303:1317 arg2 N79A 1325:1328 N79A 1325:1328 N79A 1325:1328 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 9 18 gly N-glycosylation 1303:1317 arg2 site 1319:1322 the putative N-glycosylation site 1290:1322 the putative N-glycosylation site (N79A) 1290:1329 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 11 19 theme structure 1527:1535 arg1 model 1508:1512 A model 1506:1512 A model of human TGH structure 1506:1535 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 1 20 theme microsomal 188:197 arg1 hydrolase 173:181 Triacylglycerol hydrolase 157:181 Triacylglycerol hydrolase 157:181 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 1 20 theme microsomal 188:197 arg1 enzyme 199:204 a microsomal enzyme 186:204 a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins 186:379 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 0 21 theme triad 125:129 arg1 identification 93:106 identification 93:106 identification of the catalytic triad 93:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 21 theme triad 125:129 arg1 site 151:154 a glycosylation site 135:154 a glycosylation site 135:154 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 21 theme triad 125:129 arg1 triad 125:129 the catalytic triad 111:129 the catalytic triad 111:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 11 22 theme disulfide 1616:1624 arg1 C274-C285 1648:1656 C274-C285 1648:1656 C274-C285 1648:1656 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 22 theme disulfide 1616:1624 arg1 C87-C116 1635:1642 C87-C116 1635:1642 C87-C116 1635:1642 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 22 theme disulfide 1616:1624 arg1 bridges 1626:1632 two disulfide bridges 1612:1632 two disulfide bridges (C87-C116 and C274-C285) 1612:1657 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 4 23 theme signal 781:786 arg1 sequence 788:795 the N-terminal signal sequence 766:795 the N-terminal signal sequence 766:795 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 6 24 theme Site-directed 970:982 arg1 mutagenesis 984:994 Site-directed mutagenesis 970:994 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) 970:1036 Site-directed mutagenesis of the residues (S221A, E354A, and H468A) yielded a catalytically inactive enzyme. 12022871 7 25 theme wild-type 1151:1159 arg1 enzyme 1161:1166 the wild-type enzyme 1147:1166 the wild-type enzyme 1147:1166 CD spectra of purified mutant proteins were very similar to that of the wild-type enzyme, which suggests that the mutations did not affect folding. 12022871 11 26 dep bridges 1626:1632 arg1 C274-C285 1648:1656 C274-C285 1648:1656 C274-C285 1648:1656 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 26 dep bridges 1626:1632 arg1 C87-C116 1635:1642 C87-C116 1635:1642 C87-C116 1635:1642 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 26 dep bridges 1626:1632 arg1 bridges 1626:1632 two disulfide bridges 1612:1632 two disulfide bridges (C87-C116 and C274-C285) 1612:1657 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 3 27 theme human 521:525 arg1 TGH 527:529 human TGH 521:529 human TGH 521:529 Expression of human TGH in Escherichia coli yields a protein without enzymatic activity, which suggests that posttranslational processing is necessary for the catalytic activity. 12022871 0 28 theme Structure-function 0:17 arg1 analysis 19:26 Structure-function analysis 0:26 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 0:155 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 11 29 theme active 1596:1601 arg1 site 1603:1606 a buried active site 1587:1606 a buried active site 1587:1606 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 30 theme human 1517:1521 arg1 structure 1527:1535 human TGH structure 1517:1535 human TGH structure 1517:1535 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 2 31 theme site-directed 452:464 arg1 mutagenesis 466:476 site-directed mutagenesis 452:476 site-directed mutagenesis 452:476 The structure-activity relationship of the enzyme was investigated by site-directed mutagenesis and heterologous expression. 12022871 0 32 theme human 31:35 arg1 hydrolase 53:61 human triacylglycerol hydrolase 31:61 human triacylglycerol hydrolase 31:61 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 33 theme glycosylation 137:149 arg1 site 151:154 a glycosylation site 135:154 a glycosylation site 135:154 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 33 theme glycosylation 137:149 arg1 triad 125:129 the catalytic triad 111:129 the catalytic triad 111:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 11 34 theme buried 1589:1594 arg1 site 1603:1606 a buried active site 1587:1606 a buried active site 1587:1606 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 5 35 theme glutamic 911:918 arg1 E354 926:929 E354 926:929 E354 926:929 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) was identified. 12022871 5 35 theme glutamic 911:918 arg1 acid 920:923 glutamic acid 911:923 glutamic acid (E354) 911:930 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) was identified. 12022871 11 36 theme TGH 1523:1525 arg1 structure 1527:1535 human TGH structure 1517:1535 human TGH structure 1517:1535 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 1 37 theme stored 222:227 arg1 triacylglycerol 241:255 stored cytoplasmic triacylglycerol 222:255 stored cytoplasmic triacylglycerol in the liver 222:268 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 11 38 with fold 1577:1580 arg1 C274-C285 1648:1656 C274-C285 1648:1656 C274-C285 1648:1656 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 38 with fold 1577:1580 arg1 site 1603:1606 a buried active site 1587:1606 a buried active site 1587:1606 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 38 with fold 1577:1580 arg1 C87-C116 1635:1642 C87-C116 1635:1642 C87-C116 1635:1642 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 11 38 with fold 1577:1580 arg1 bridges 1626:1632 two disulfide bridges 1612:1632 two disulfide bridges (C87-C116 and C274-C285) 1612:1657 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 3 39 theme catalytic 666:674 arg1 activity 676:683 the catalytic activity 662:683 the catalytic activity 662:683 Expression of human TGH in Escherichia coli yields a protein without enzymatic activity, which suggests that posttranslational processing is necessary for the catalytic activity. 12022871 1 40 theme cytoplasmic 229:239 arg1 triacylglycerol 241:255 stored cytoplasmic triacylglycerol 222:255 stored cytoplasmic triacylglycerol in the liver 222:268 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 0 41 theme hydrolase 53:61 arg1 analysis 19:26 Structure-function analysis 0:26 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 0:155 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 5 42 theme putative 842:849 arg1 triad 861:865 A putative catalytic triad 840:865 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) 840:952 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) was identified. 12022871 9 43 theme putative 1294:1301 arg1 N79A 1325:1328 N79A 1325:1328 N79A 1325:1328 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 9 43 theme putative 1294:1301 arg1 site 1319:1322 the putative N-glycosylation site 1290:1322 the putative N-glycosylation site (N79A) 1290:1329 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 3 44 theme TGH 527:529 arg1 Expression 507:516 Expression 507:516 Expression of human TGH in Escherichia coli 507:549 Expression of human TGH in Escherichia coli yields a protein without enzymatic activity, which suggests that posttranslational processing is necessary for the catalytic activity. 12022871 4 45 theme N-terminal 770:779 arg1 sequence 788:795 the N-terminal signal sequence 766:795 the N-terminal signal sequence 766:795 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 2 46 theme heterologous 482:493 arg1 expression 495:504 heterologous expression 482:504 heterologous expression 482:504 The structure-activity relationship of the enzyme was investigated by site-directed mutagenesis and heterologous expression. 12022871 1 47 theme very-low-density 351:366 arg1 lipoproteins 368:379 very-low-density lipoproteins 351:379 very-low-density lipoproteins 351:379 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 0 48 theme triacylglycerol 37:51 arg1 hydrolase 53:61 human triacylglycerol hydrolase 31:61 human triacylglycerol hydrolase 31:61 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 9 49 theme N-glycosylation 1303:1317 arg1 N79A 1325:1328 N79A 1325:1328 N79A 1325:1328 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 9 49 theme N-glycosylation 1303:1317 arg1 site 1319:1322 the putative N-glycosylation site 1290:1322 the putative N-glycosylation site (N79A) 1290:1329 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 8 50 theme insect 1261:1266 arg1 cells 1268:1272 the insect cells 1257:1272 the insect cells 1257:1272 Human TGH was glycosylated in the insect cells. 12022871 1 51 from triacylglycerol 241:255 arg1 liver 264:268 the liver 260:268 the liver 260:268 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 4 52 from Expression 686:695 arg1 cells 726:730 baculovirus-infected Sf-9 cells 700:730 baculovirus-infected Sf-9 cells 700:730 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 0 53 theme site-directed 66:78 arg1 mutagenesis 80:90 site-directed mutagenesis 66:90 site-directed mutagenesis 66:90 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 1 54 theme lipoproteins 368:379 arg1 assembly 339:346 the assembly 335:346 the assembly of very-low-density lipoproteins 335:379 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 9 55 theme site 1319:1322 arg1 Mutagenesis 1275:1285 Mutagenesis 1275:1285 Mutagenesis of the putative N-glycosylation site (N79A) 1275:1329 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 10 56 theme endoplasmic 1409:1419 arg1 reticulum 1421:1429 C-terminal endoplasmic reticulum 1398:1429 the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) 1385:1453 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 7 57 theme mutant 1102:1107 arg1 proteins 1109:1116 purified mutant proteins 1093:1116 purified mutant proteins 1093:1116 CD spectra of purified mutant proteins were very similar to that of the wild-type enzyme, which suggests that the mutations did not affect folding. 12022871 5 58 theme catalytic 851:859 arg1 triad 861:865 A putative catalytic triad 840:865 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) 840:952 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) was identified. 12022871 10 59 theme C-terminal 1398:1407 arg1 reticulum 1421:1429 C-terminal endoplasmic reticulum 1398:1429 the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) 1385:1453 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 0 60 gly glycosylation 137:149 arg2 triad 125:129 the catalytic triad 111:129 the catalytic triad 111:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 60 gly glycosylation 137:149 arg1 triad 125:129 the catalytic triad 111:129 the catalytic triad 111:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 60 gly glycosylation 137:149 arg2 site 151:154 a glycosylation site 135:154 a glycosylation site 135:154 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 7 61 theme proteins 1109:1116 arg1 spectra 1082:1088 CD spectra 1079:1088 CD spectra of purified mutant proteins 1079:1116 CD spectra of purified mutant proteins were very similar to that of the wild-type enzyme, which suggests that the mutations did not affect folding. 12022871 10 62 theme putative 1389:1396 arg1 signal 1441:1446 the putative C-terminal endoplasmic reticulum retrieval signal 1385:1446 the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) 1385:1453 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 10 62 theme putative 1389:1396 arg1 HIEL 1449:1452 HIEL 1449:1452 HIEL 1449:1452 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 11 63 theme lipase 1549:1554 arg1 fold 1577:1580 a lipase alpha/beta hydrolase fold 1547:1580 a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285) 1547:1657 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 8 64 theme Human 1227:1231 arg1 TGH 1233:1235 Human TGH 1227:1235 Human TGH 1227:1235 Human TGH was glycosylated in the insect cells. 12022871 7 65 theme purified 1093:1100 arg1 proteins 1109:1116 purified mutant proteins 1093:1116 purified mutant proteins 1093:1116 CD spectra of purified mutant proteins were very similar to that of the wild-type enzyme, which suggests that the mutations did not affect folding. 12022871 2 66 theme structure-activity 386:403 arg1 relationship 405:416 The structure-activity relationship 382:416 The structure-activity relationship of the enzyme 382:430 The structure-activity relationship of the enzyme was investigated by site-directed mutagenesis and heterologous expression. 12022871 10 67 theme reticulum 1421:1429 arg1 signal 1441:1446 the putative C-terminal endoplasmic reticulum retrieval signal 1385:1446 the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) 1385:1453 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 10 67 theme reticulum 1421:1429 arg1 HIEL 1449:1452 HIEL 1449:1452 HIEL 1449:1452 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 5 68 theme nucleophilic 883:894 arg1 serine 896:901 a nucleophilic serine 881:901 a nucleophilic serine (S221) 881:908 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) was identified. 12022871 5 68 theme nucleophilic 883:894 arg1 S221 904:907 S221 904:907 S221 904:907 A putative catalytic triad consisting of a nucleophilic serine (S221), glutamic acid (E354), and histidine (H468) was identified. 12022871 11 69 theme hydrolase 1567:1575 arg1 fold 1577:1580 a lipase alpha/beta hydrolase fold 1547:1580 a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285) 1547:1657 A model of human TGH structure suggested a lipase alpha/beta hydrolase fold with a buried active site and two disulfide bridges (C87-C116 and C274-C285). 12022871 0 70 dep analysis 19:26 arg1 site 151:154 a glycosylation site 135:154 a glycosylation site 135:154 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 70 dep analysis 19:26 arg1 identification 93:106 identification 93:106 identification of the catalytic triad 93:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 0 70 dep analysis 19:26 arg1 triad 125:129 the catalytic triad 111:129 the catalytic triad 111:129 Structure-function analysis of human triacylglycerol hydrolase by site-directed mutagenesis: identification of the catalytic triad and a glycosylation site. 12022871 7 71 theme CD 1079:1080 arg1 spectra 1082:1088 CD spectra 1079:1088 CD spectra of purified mutant proteins 1079:1116 CD spectra of purified mutant proteins were very similar to that of the wild-type enzyme, which suggests that the mutations did not affect folding. 12022871 1 72 theme Triacylglycerol 157:171 arg1 hydrolase 173:181 Triacylglycerol hydrolase 157:181 Triacylglycerol hydrolase 157:181 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 1 72 theme Triacylglycerol 157:171 arg1 enzyme 199:204 a microsomal enzyme 186:204 a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins 186:379 Triacylglycerol hydrolase is a microsomal enzyme that hydrolyzes stored cytoplasmic triacylglycerol in the liver and participates in the lipolysis/re-esterification cycle during the assembly of very-low-density lipoproteins. 12022871 4 73 theme correct 744:750 arg1 processing 752:761 correct processing 744:761 correct processing of the N-terminal signal sequence 744:795 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 4 74 theme Sf-9 721:724 arg1 cells 726:730 baculovirus-infected Sf-9 cells 700:730 baculovirus-infected Sf-9 cells 700:730 Expression in baculovirus-infected Sf-9 cells resulted in correct processing of the N-terminal signal sequence and yielded a catalytically active enzyme. 12022871 9 75 theme nonglycosylated 1349:1363 arg1 enzyme 1365:1370 an active nonglycosylated enzyme 1339:1370 an active nonglycosylated enzyme 1339:1370 Mutagenesis of the putative N-glycosylation site (N79A) yielded an active nonglycosylated enzyme. 12022871 10 76 theme signal 1441:1446 arg1 Deletion 1373:1380 Deletion 1373:1380 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) 1373:1453 Deletion of the putative C-terminal endoplasmic reticulum retrieval signal (HIEL) did not result in secretion of the mutant protein. 12022871 3 77 theme posttranslational 616:632 arg1 processing 634:643 posttranslational processing 616:643 posttranslational processing 616:643 Expression of human TGH in Escherichia coli yields a protein without enzymatic activity, which suggests that posttranslational processing is necessary for the catalytic activity. 16040958 0 0 theme amino-terminal 78:91 arg1 inhibitor 110:118 the c1 inhibitor 103:118 the c1 inhibitor 103:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 0 theme amino-terminal 78:91 arg1 domain 93:98 the amino-terminal domain 74:98 the amino-terminal domain of the c1 inhibitor 74:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 1 1 theme endotoxin 321:329 arg1 shock 331:335 endotoxin shock 321:335 endotoxin shock 321:335 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 4 2 from residues 959:966 arg1 mutation 899:906 The mutation 895:906 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) 895:1007 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 3 3 theme Asn 846:848 arg1 substitution 830:841 substitution 830:841 substitution of Asn(47) or Asn(59) 830:863 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 1 4 theme C1 250:251 arg1 inhibitor 253:261 The C1 inhibitor 246:261 The C1 inhibitor (C1INH) 246:269 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 4 theme C1 250:251 arg1 protein 303:309 a plasma complement regulatory protein 272:309 a plasma complement regulatory protein 272:309 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 4 theme C1 250:251 arg1 C1INH 264:268 C1INH 264:268 C1INH 264:268 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 3 5 from effective 791:799 arg1 binding 808:814 its binding 804:814 its binding to LPS 804:821 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 2 6 theme glycosylation 650:662 arg1 sites 664:668 the three N-linked glycosylation sites 631:668 the three N-linked glycosylation sites 631:668 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 0 7 theme Inhibitor 159:167 arg1 interaction 137:147 interaction 137:147 interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A 137:243 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 1 8 theme nonserpin 427:435 arg1 region 437:442 its amino-terminal heavily glycosylated nonserpin region 387:442 its amino-terminal heavily glycosylated nonserpin region 387:442 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 9 gly glycosylated 414:425 arg1 region 437:442 its amino-terminal heavily glycosylated nonserpin region 387:442 its amino-terminal heavily glycosylated nonserpin region 387:442 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 2 10 theme LPS-binding 537:547 arg1 s 554:554 the potential LPS-binding site(s) 523:555 the potential LPS-binding site(s) within the amino-terminal domain 523:588 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 5 11 from binding 1134:1140 arg1 macrophages 1231:1241 RAW 264.7 macrophages 1221:1241 RAW 264.7 macrophages 1221:1241 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 0 12 theme lipid 237:241 arg1 A 243:243 lipid A 237:243 lipid A 237:243 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 7 13 from Asn 1499:1501 arg1 carbohydrate 1483:1494 the N-linked carbohydrate 1470:1494 the N-linked carbohydrate at Asn(3) 1470:1504 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 0 14 with interaction 137:147 arg1 lipopolysaccharide 214:231 Salmonella enterica serovar typhimurium lipopolysaccharide 174:231 Salmonella enterica serovar typhimurium lipopolysaccharide 174:231 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 14 with interaction 137:147 arg1 A 243:243 lipid A 237:243 lipid A 237:243 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 2 15 theme potential 527:535 arg1 s 554:554 the potential LPS-binding site(s) 523:555 the potential LPS-binding site(s) within the amino-terminal domain 523:588 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 5 16 theme factor 1189:1194 arg1 alpha 1196:1200 tumor necrosis factor alpha 1174:1200 tumor necrosis factor alpha mRNA expression 1174:1216 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 5 17 theme necrosis 1180:1187 arg1 alpha 1196:1200 tumor necrosis factor alpha 1174:1200 tumor necrosis factor alpha mRNA expression 1174:1216 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 2 18 theme N-linked 641:648 arg1 sites 664:668 the three N-linked glycosylation sites 631:668 the three N-linked glycosylation sites 631:668 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 6 19 theme lipid 1302:1306 arg1 A 1308:1308 diphosphoryl lipid A 1289:1308 diphosphoryl lipid A 1289:1308 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 0 20 theme c1 107:108 arg1 inhibitor 110:118 the c1 inhibitor 103:118 the c1 inhibitor 103:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 7 21 link N-linked 1474:1481 arg1 carbohydrate 1483:1494 the N-linked carbohydrate 1470:1494 the N-linked carbohydrate at Asn(3) 1470:1504 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 0 22 theme typhimurium 202:212 arg1 lipopolysaccharide 214:231 Salmonella enterica serovar typhimurium lipopolysaccharide 174:231 Salmonella enterica serovar typhimurium lipopolysaccharide 174:231 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 23 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation at Asn3 0:29 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 7 24 with interaction 1395:1405 arg1 LPS 1445:1447 gram-negative bacterial LPS 1421:1447 gram-negative bacterial LPS 1421:1447 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 5 25 from up-regulation 1157:1169 arg1 macrophages 1231:1241 RAW 264.7 macrophages 1221:1241 RAW 264.7 macrophages 1221:1241 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 2 26 gly glycosylation 650:662 arg2 three 635:639 three 635:639 three 635:639 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 2 26 gly glycosylation 650:662 arg2 sites 664:668 the three N-linked glycosylation sites 631:668 the three N-linked glycosylation sites 631:668 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 1 27 theme bacterial 463:471 arg1 lipopolysaccharide 473:490 gram-negative bacterial lipopolysaccharide 449:490 gram-negative bacterial lipopolysaccharide (LPS) 449:496 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 27 theme bacterial 463:471 arg1 LPS 493:495 LPS 493:495 LPS 493:495 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 5 28 theme alpha 1196:1200 arg1 expression 1207:1216 tumor necrosis factor alpha mRNA expression 1174:1216 tumor necrosis factor alpha mRNA expression 1174:1216 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 5 29 theme LPS 1130:1132 arg1 binding 1134:1140 LPS binding 1130:1140 LPS binding 1130:1140 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 2 30 link N-linked 641:648 arg1 sites 664:668 the three N-linked glycosylation sites 631:668 the three N-linked glycosylation sites 631:668 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 7 31 theme gram-negative 1421:1433 arg1 LPS 1445:1447 gram-negative bacterial LPS 1421:1447 gram-negative bacterial LPS 1421:1447 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 5 32 theme expression 1207:1216 arg1 up-regulation 1157:1169 LPS-induced up-regulation 1145:1169 LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages 1145:1241 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 5 32 theme expression 1207:1216 arg1 binding 1134:1140 LPS binding 1130:1140 LPS binding 1130:1140 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 3 33 from binding 808:814 arg1 effective 791:799 effective 791:799 effective 791:799 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 3 33 from binding 808:814 arg1 mutant 728:733 A mutant 726:733 A mutant in which Asn(3) was replaced with Ala 726:771 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 7 34 theme N-linked 1474:1481 arg1 carbohydrate 1483:1494 the N-linked carbohydrate 1470:1494 the N-linked carbohydrate at Asn(3) 1470:1504 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 0 35 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation at Asn3 0:29 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 7 36 theme bacterial 1435:1443 arg1 LPS 1445:1447 gram-negative bacterial LPS 1421:1447 gram-negative bacterial LPS 1421:1447 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 7 37 theme charged 1528:1534 arg1 residues 1536:1543 the positively charged residues 1513:1543 the positively charged residues within the amino-terminal domain 1513:1576 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 1 38 with interaction 372:382 arg1 lipopolysaccharide 473:490 gram-negative bacterial lipopolysaccharide 449:490 gram-negative bacterial lipopolysaccharide (LPS) 449:496 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 38 with interaction 372:382 arg1 LPS 493:495 LPS 493:495 LPS 493:495 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 6 39 theme recombinant 1351:1361 arg1 C1INH 1373:1377 recombinant wild-type C1INH 1351:1377 recombinant wild-type C1INH 1351:1377 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 4 40 theme amino 948:952 arg1 Lys 1000:1002 Lys 1000:1002 Lys 1000:1002 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 40 theme amino 948:952 arg1 Lys 987:989 Lys 987:989 Lys 987:989 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 40 theme amino 948:952 arg1 Lys 978:980 Lys 978:980 Lys 978:980 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 40 theme amino 948:952 arg1 Arg 969:971 Arg(18) 969:975 Arg(18) 969:975 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 40 theme amino 948:952 arg1 residues 959:966 all four positively charged amino acid residues 920:966 all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) 920:1007 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 5 41 theme RAW 1221:1223 arg1 macrophages 1231:1241 RAW 264.7 macrophages 1221:1241 RAW 264.7 macrophages 1221:1241 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 1 42 theme gram-negative 449:461 arg1 lipopolysaccharide 473:490 gram-negative bacterial lipopolysaccharide 449:490 gram-negative bacterial lipopolysaccharide (LPS) 449:496 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 42 theme gram-negative 449:461 arg1 LPS 493:495 LPS 493:495 LPS 493:495 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 4 43 theme charged 940:946 arg1 Lys 1000:1002 Lys 1000:1002 Lys 1000:1002 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 43 theme charged 940:946 arg1 Lys 987:989 Lys 987:989 Lys 987:989 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 43 theme charged 940:946 arg1 Lys 978:980 Lys 978:980 Lys 978:980 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 43 theme charged 940:946 arg1 Arg 969:971 Arg(18) 969:975 Arg(18) 969:975 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 43 theme charged 940:946 arg1 residues 959:966 all four positively charged amino acid residues 920:966 all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) 920:1007 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 0 44 theme inhibitor 110:118 arg1 inhibitor 110:118 the c1 inhibitor 103:118 the c1 inhibitor 103:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 44 theme inhibitor 110:118 arg1 domain 93:98 the amino-terminal domain 74:98 the amino-terminal domain of the c1 inhibitor 74:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 5 45 theme mRNA 1202:1205 arg1 expression 1207:1216 tumor necrosis factor alpha mRNA expression 1174:1216 tumor necrosis factor alpha mRNA expression 1174:1216 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 3 46 theme Asn 857:859 arg1 substitution 830:841 substitution 830:841 substitution of Asn(47) or Asn(59) 830:863 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 4 47 theme C1INH 911:915 arg1 mutation 899:906 The mutation 895:906 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) 895:1007 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 1 48 theme plasma 274:279 arg1 inhibitor 253:261 The C1 inhibitor 246:261 The C1 inhibitor (C1INH) 246:269 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 48 theme plasma 274:279 arg1 protein 303:309 a plasma complement regulatory protein 272:309 a plasma complement regulatory protein 272:309 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 49 theme direct 365:370 arg1 interaction 372:382 the direct interaction 361:382 the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS) 361:496 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 6 50 theme diphosphoryl 1289:1300 arg1 A 1308:1308 diphosphoryl lipid A 1289:1308 diphosphoryl lipid A 1289:1308 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 5 51 theme C1INH 1069:1073 arg1 mutants 1075:1081 The C1INH mutants 1065:1081 The C1INH mutants that did not bind to LPS 1065:1106 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 1 52 theme complement 281:290 arg1 inhibitor 253:261 The C1 inhibitor 246:261 The C1 inhibitor (C1INH) 246:269 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 52 theme complement 281:290 arg1 protein 303:309 a plasma complement regulatory protein 272:309 a plasma complement regulatory protein 272:309 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 2 53 theme site 549:552 arg1 s 554:554 the potential LPS-binding site(s) 523:555 the potential LPS-binding site(s) within the amino-terminal domain 523:588 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 0 54 gly glycosylation 9:21 arg1 Inhibitor 159:167 the C1 Inhibitor 152:167 the C1 Inhibitor 152:167 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 54 gly glycosylation 9:21 arg1 Asn3 26:29 Asn3 26:29 Asn3 26:29 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 54 gly glycosylation 9:21 arg2 Asn3 26:29 Asn3 26:29 Asn3 26:29 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 2 55 theme acid 711:714 arg1 residues 716:723 the four positively charged amino acid residues 677:723 the four positively charged amino acid residues 677:723 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 0 56 theme charged 50:56 arg1 residues 58:65 the positively charged residues 35:65 the positively charged residues within the amino-terminal domain of the c1 inhibitor 35:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 1 57 theme regulatory 292:301 arg1 inhibitor 253:261 The C1 inhibitor 246:261 The C1 inhibitor (C1INH) 246:269 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 57 theme regulatory 292:301 arg1 protein 303:309 a plasma complement regulatory protein 272:309 a plasma complement regulatory protein 272:309 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 7 58 theme C1INH 1410:1414 arg1 interaction 1395:1405 the interaction 1391:1405 the interaction of C1INH with gram-negative bacterial LPS 1391:1447 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 7 58 theme C1INH 1410:1414 arg1 dependent 1452:1460 dependent 1452:1460 dependent 1452:1460 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 2 59 theme amino 705:709 arg1 residues 716:723 the four positively charged amino acid residues 677:723 the four positively charged amino acid residues 677:723 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 4 60 dep residues 959:966 arg1 Lys 987:989 Lys 987:989 Lys 987:989 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 residues 959:966 all four positively charged amino acid residues 920:966 all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) 920:1007 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 30 991:992 30 991:992 30 991:992 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 55 1004:1005 55 1004:1005 55 1004:1005 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 Lys 978:980 Lys 978:980 Lys 978:980 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 Arg 969:971 Arg(18) 969:975 Arg(18) 969:975 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 22 982:983 22 982:983 22 982:983 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 60 dep residues 959:966 arg1 Lys 1000:1002 Lys 1000:1002 Lys 1000:1002 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 6 61 with comparison 1327:1336 arg1 that 1343:1346 that 1343:1346 that 1343:1346 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 2 62 theme amino-terminal 568:581 arg1 domain 583:588 the amino-terminal domain 564:588 the amino-terminal domain 564:588 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 2 63 theme charged 697:703 arg1 residues 716:723 the four positively charged amino acid residues 677:723 the four positively charged amino acid residues 677:723 To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. 16040958 4 64 theme near-complete 1021:1033 arg1 failure 1035:1041 near-complete failure 1021:1041 near-complete failure to interact with LPS 1021:1062 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 0 65 theme C1 156:157 arg1 Inhibitor 159:167 the C1 Inhibitor 152:167 the C1 Inhibitor 152:167 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 1 66 theme amino-terminal 391:404 arg1 region 437:442 its amino-terminal heavily glycosylated nonserpin region 387:442 its amino-terminal heavily glycosylated nonserpin region 387:442 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 1 67 theme region 437:442 arg1 interaction 372:382 the direct interaction 361:382 the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS) 361:496 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 6 68 theme mutants 1278:1284 arg1 binding 1261:1267 the binding 1257:1267 the binding of C1INH mutants to diphosphoryl lipid A 1257:1308 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 3 69 theme little 869:874 arg1 effect 876:881 little effect 869:881 little effect 869:881 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 4 70 theme acid 954:957 arg1 Lys 1000:1002 Lys 1000:1002 Lys 1000:1002 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 70 theme acid 954:957 arg1 Lys 987:989 Lys 987:989 Lys 987:989 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 70 theme acid 954:957 arg1 Lys 978:980 Lys 978:980 Lys 978:980 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 70 theme acid 954:957 arg1 Arg 969:971 Arg(18) 969:975 Arg(18) 969:975 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 4 70 theme acid 954:957 arg1 residues 959:966 all four positively charged amino acid residues 920:966 all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) 920:1007 The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near-complete failure to interact with LPS. 16040958 5 71 theme tumor 1174:1178 arg1 alpha 1196:1200 tumor necrosis factor alpha 1174:1200 tumor necrosis factor alpha mRNA expression 1174:1216 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 6 72 theme wild-type 1363:1371 arg1 C1INH 1373:1377 recombinant wild-type C1INH 1351:1377 recombinant wild-type C1INH 1351:1377 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 6 73 theme C1INH 1272:1276 arg1 mutants 1278:1284 C1INH mutants 1272:1284 C1INH mutants 1272:1284 In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. 16040958 1 74 theme glycosylated 414:425 arg1 region 437:442 its amino-terminal heavily glycosylated nonserpin region 387:442 its amino-terminal heavily glycosylated nonserpin region 387:442 The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino-terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). 16040958 5 75 theme LPS-induced 1145:1155 arg1 up-regulation 1157:1169 LPS-induced up-regulation 1145:1169 LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages 1145:1241 The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. 16040958 7 76 theme amino-terminal 1556:1569 arg1 domain 1571:1576 the amino-terminal domain 1552:1576 the amino-terminal domain 1552:1576 Therefore, the interaction of C1INH with gram-negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. 16040958 0 77 from Asn3 26:29 arg1 residues 58:65 the positively charged residues 35:65 the positively charged residues within the amino-terminal domain of the c1 inhibitor 35:118 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 0 77 from Asn3 26:29 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation at Asn3 0:29 N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. 16040958 3 78 contain had 865:867 arg1 substitution 830:841 substitution 830:841 substitution of Asn(47) or Asn(59) 830:863 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 16040958 3 78 contain had 865:867 arg2 effect 876:881 little effect 869:881 little effect 869:881 A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. 27033522 11 0 theme PEBP4 1511:1515 arg1 siRNA 1517:1521 PEBP4 siRNA 1511:1521 PEBP4 siRNA 1511:1521 In contrast, PEBP4 siRNA suppressed phosphorylation of Act at S473. 27033522 3 1 theme glycosylation 483:495 arg1 sites 497:501 potential glycosylation sites 473:501 potential glycosylation sites 473:501 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 9 2 theme epitope 1300:1306 arg1 tag 1308:1310 a C-terminal epitope tag 1287:1310 a C-terminal epitope tag 1287:1310 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 3 3 theme functional 303:312 arg1 differences 314:324 the functional differences 299:324 the functional differences among PEBP1-4 299:338 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 5 4 dep both 747:750 arg1 mouse 752:756 mouse 752:756 mouse 752:756 Our data revealed that both mouse and human PEBP4 were secreted when the epitope was tagged at their C-terminus. 27033522 12 5 theme multifunctional 1613:1627 arg1 PEBP4 1602:1606 PEBP4 1602:1606 PEBP4 1602:1606 Therefore, our results suggest that PEBP4 is a multifunctional protein and can be secreted. 27033522 12 5 theme multifunctional 1613:1627 arg1 protein 1629:1635 a multifunctional protein 1611:1635 a multifunctional protein 1611:1635 Therefore, our results suggest that PEBP4 is a multifunctional protein and can be secreted. 27033522 4 6 theme carboxyl 643:650 arg1 terminus 656:663 carboxyl (C) terminus 643:663 carboxyl (C) terminus 643:663 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 0 7 theme multiple 81:88 arg1 functions 90:98 multiple functions 81:98 multiple functions 81:98 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 9 8 theme N-terminal 1201:1210 arg1 PEBP4 1219:1223 N-terminal tagged PEBP4 1201:1223 N-terminal tagged PEBP4 1201:1223 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 3 9 theme intracellular 524:536 arg1 PEBP1-3 512:518 PEBP1-3 512:518 PEBP1-3 512:518 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 3 9 theme intracellular 524:536 arg1 proteins 538:545 intracellular proteins 524:545 intracellular proteins 524:545 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 13 10 theme cellular 1749:1756 arg1 events 1758:1763 cellular events 1749:1763 cellular events 1749:1763 It will be important to investigate the mechanism by which PEBP4 is secreted and regulates cellular events. 27033522 6 11 theme conserved 898:906 arg1 domain 908:913 the C-terminal conserved domain 883:913 the C-terminal conserved domain in addition to the N-terminal signal sequence 883:959 To our surprise, secretion was dependent upon the C-terminal conserved domain in addition to the N-terminal signal sequence. 27033522 3 12 theme underlying 348:357 arg1 mechanism 359:367 the underlying mechanism 344:367 the underlying mechanism for their actions 344:385 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 9 13 theme tagged 1212:1217 arg1 PEBP4 1219:1223 N-terminal tagged PEBP4 1201:1223 N-terminal tagged PEBP4 1201:1223 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 7 14 theme recombinant 1013:1023 arg1 protein 1025:1031 the recombinant protein 1009:1031 the recombinant protein 1009:1031 When the epitope was placed to the N-terminus, the recombinant protein failed to secrete and instead, was retained in the cytoplasm. 27033522 6 15 theme C-terminal 887:896 arg1 domain 908:913 the C-terminal conserved domain 883:913 the C-terminal conserved domain in addition to the N-terminal signal sequence 883:959 To our surprise, secretion was dependent upon the C-terminal conserved domain in addition to the N-terminal signal sequence. 27033522 9 16 theme PEBP4 1219:1223 arg1 overexpression 1183:1196 overexpression 1183:1196 overexpression of N-terminal tagged PEBP4 1183:1223 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 1 17 theme proteins 177:184 arg1 superfamily 162:172 a superfamily 160:172 a superfamily of proteins that are conserved from bacteria to humans 160:227 Phosphatidylethanolamine binding proteins (PEBP) represent a superfamily of proteins that are conserved from bacteria to humans. 27033522 9 18 contain have 1320:1323 arg1 that 1277:1280 that 1277:1280 that 1277:1280 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 9 18 contain have 1320:1323 arg2 effect 1333:1338 such an effect 1325:1338 such an effect 1325:1338 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 3 19 gly glycosylation 483:495 arg2 sites 497:501 potential glycosylation sites 473:501 potential glycosylation sites 473:501 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 4 20 theme amino 621:625 arg1 terminus 631:638 amino (N) terminus 621:638 amino (N) terminus 621:638 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 0 21 theme binding 25:31 arg1 protein 33:39 Phosphatidylethanolamine binding protein 4 0:41 Phosphatidylethanolamine binding protein 4 (PEBP4) 0:49 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 21 theme binding 25:31 arg1 PEBP4 44:48 PEBP4 44:48 PEBP4 44:48 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 21 theme binding 25:31 arg1 protein 65:71 a secreted protein 54:71 a secreted protein 54:71 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 9 22 theme ERK 1254:1256 arg1 activation 1258:1267 ERK activation 1254:1267 ERK activation 1254:1267 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 3 23 theme sequence 403:410 arg1 alignment 412:420 a sequence alignment 401:420 a sequence alignment 401:420 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 0 24 theme Phosphatidylethanolamine 0:23 arg1 protein 33:39 Phosphatidylethanolamine binding protein 4 0:41 Phosphatidylethanolamine binding protein 4 (PEBP4) 0:49 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 24 theme Phosphatidylethanolamine 0:23 arg1 PEBP4 44:48 PEBP4 44:48 PEBP4 44:48 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 24 theme Phosphatidylethanolamine 0:23 arg1 protein 65:71 a secreted protein 54:71 a secreted protein 54:71 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 4 25 theme N 628:628 arg1 terminus 631:638 amino (N) terminus 621:638 amino (N) terminus 621:638 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 8 26 theme Mass 1095:1098 arg1 spectrometry 1100:1111 Mass spectrometry 1095:1111 Mass spectrometry 1095:1111 Mass spectrometry detected asparagine (N)-glycosylation on the secreted PEBP4. 27033522 8 27 theme secreted 1158:1165 arg1 PEBP4 1167:1171 the secreted PEBP4 1154:1171 the secreted PEBP4 1154:1171 Mass spectrometry detected asparagine (N)-glycosylation on the secreted PEBP4. 27033522 7 28 dep failed 1033:1038 arg1 retained 1068:1075 retained 1068:1075 was retained in the cytoplasm 1064:1092 When the epitope was placed to the N-terminus, the recombinant protein failed to secrete and instead, was retained in the cytoplasm. 27033522 10 29 theme shRNA 1373:1377 arg1 transfection 1351:1362 transfection 1351:1362 transfection of PEBP4 shRNA 1351:1377 Likewise, transfection of PEBP4 shRNA did not appear to affect ERK activation, suggesting that PEBP4 does not participate in the regulation of this pathway. 27033522 4 30 theme signal 677:682 arg1 sequence 684:691 the signal sequence 673:691 the signal sequence 673:691 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 4 31 theme Myc 602:604 arg1 epitope 606:612 Myc epitope 602:612 Myc epitope 602:612 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 3 32 theme potential 473:481 arg1 sites 497:501 potential glycosylation sites 473:501 potential glycosylation sites 473:501 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 10 33 theme PEBP4 1367:1371 arg1 shRNA 1373:1377 PEBP4 shRNA 1367:1377 PEBP4 shRNA 1367:1377 Likewise, transfection of PEBP4 shRNA did not appear to affect ERK activation, suggesting that PEBP4 does not participate in the regulation of this pathway. 27033522 10 34 theme pathway 1489:1495 arg1 regulation 1470:1479 the regulation 1466:1479 the regulation of this pathway 1466:1495 Likewise, transfection of PEBP4 shRNA did not appear to affect ERK activation, suggesting that PEBP4 does not participate in the regulation of this pathway. 27033522 9 35 theme activation 1258:1267 arg1 inhibition 1240:1249 an inhibition 1237:1249 an inhibition of ERK activation by EGF 1237:1274 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 8 36 gly -glycosylation 1136:1149 arg1 PEBP4 1167:1171 the secreted PEBP4 1154:1171 the secreted PEBP4 1154:1171 Mass spectrometry detected asparagine (N)-glycosylation on the secreted PEBP4. 27033522 4 37 with constructs 586:595 arg1 epitope 606:612 Myc epitope 602:612 Myc epitope 602:612 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 6 38 theme signal 945:950 arg1 sequence 952:959 the N-terminal signal sequence 930:959 the N-terminal signal sequence 930:959 To our surprise, secretion was dependent upon the C-terminal conserved domain in addition to the N-terminal signal sequence. 27033522 3 39 contain contains 443:450 arg1 PEBP4 437:441 PEBP4 437:441 PEBP4 437:441 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 3 39 contain contains 443:450 arg2 sites 497:501 potential glycosylation sites 473:501 potential glycosylation sites 473:501 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 3 39 contain contains 443:450 arg2 peptide 461:467 a signal peptide 452:467 a signal peptide 452:467 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 1 40 theme Phosphatidylethanolamine 101:124 arg1 proteins 134:141 Phosphatidylethanolamine binding proteins 101:141 Phosphatidylethanolamine binding proteins (PEBP) 101:148 Phosphatidylethanolamine binding proteins (PEBP) represent a superfamily of proteins that are conserved from bacteria to humans. 27033522 1 40 theme Phosphatidylethanolamine 101:124 arg1 PEBP 144:147 PEBP 144:147 PEBP 144:147 Phosphatidylethanolamine binding proteins (PEBP) represent a superfamily of proteins that are conserved from bacteria to humans. 27033522 8 41 theme asparagine 1122:1131 arg1 -glycosylation 1136:1149 asparagine (N)-glycosylation 1122:1149 asparagine (N)-glycosylation 1122:1149 Mass spectrometry detected asparagine (N)-glycosylation on the secreted PEBP4. 27033522 0 42 contain has 77:79 arg1 protein 33:39 Phosphatidylethanolamine binding protein 4 0:41 Phosphatidylethanolamine binding protein 4 (PEBP4) 0:49 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 42 contain has 77:79 arg1 PEBP4 44:48 PEBP4 44:48 PEBP4 44:48 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 42 contain has 77:79 arg2 functions 90:98 multiple functions 81:98 multiple functions 81:98 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 42 contain has 77:79 arg1 protein 65:71 a secreted protein 54:71 a secreted protein 54:71 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 1 43 theme binding 126:132 arg1 proteins 134:141 Phosphatidylethanolamine binding proteins 101:141 Phosphatidylethanolamine binding proteins (PEBP) 101:148 Phosphatidylethanolamine binding proteins (PEBP) represent a superfamily of proteins that are conserved from bacteria to humans. 27033522 1 43 theme binding 126:132 arg1 PEBP 144:147 PEBP 144:147 PEBP 144:147 Phosphatidylethanolamine binding proteins (PEBP) represent a superfamily of proteins that are conserved from bacteria to humans. 27033522 11 44 theme Act 1553:1555 arg1 phosphorylation 1534:1548 phosphorylation 1534:1548 phosphorylation of Act 1534:1555 In contrast, PEBP4 siRNA suppressed phosphorylation of Act at S473. 27033522 3 45 theme signal 454:459 arg1 peptide 461:467 a signal peptide 452:467 a signal peptide 452:467 To determine the functional differences among PEBP1-4 and the underlying mechanism for their actions, we performed a sequence alignment and found that PEBP4 contains a signal peptide and potential glycosylation sites, whereas PEBP1-3 are intracellular proteins. 27033522 6 46 theme N-terminal 934:943 arg1 sequence 952:959 the N-terminal signal sequence 930:959 the N-terminal signal sequence 930:959 To our surprise, secretion was dependent upon the C-terminal conserved domain in addition to the N-terminal signal sequence. 27033522 4 47 dep terminus 631:638 arg1 the 617:619 the 617:619 the 617:619 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 27033522 5 48 dep PEBP4 768:772 arg1 human 762:766 human 762:766 human 762:766 Our data revealed that both mouse and human PEBP4 were secreted when the epitope was tagged at their C-terminus. 27033522 5 48 dep PEBP4 768:772 arg1 both 747:750 both 747:750 both 747:750 Our data revealed that both mouse and human PEBP4 were secreted when the epitope was tagged at their C-terminus. 27033522 0 49 theme secreted 56:63 arg1 protein 65:71 a secreted protein 54:71 a secreted protein 54:71 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 0 49 theme secreted 56:63 arg1 protein 33:39 Phosphatidylethanolamine binding protein 4 0:41 Phosphatidylethanolamine binding protein 4 (PEBP4) 0:49 Phosphatidylethanolamine binding protein 4 (PEBP4) is a secreted protein and has multiple functions. 27033522 10 50 theme ERK 1404:1406 arg1 activation 1408:1417 ERK activation 1404:1417 ERK activation 1404:1417 Likewise, transfection of PEBP4 shRNA did not appear to affect ERK activation, suggesting that PEBP4 does not participate in the regulation of this pathway. 27033522 9 51 theme C-terminal 1289:1298 arg1 tag 1308:1310 a C-terminal epitope tag 1287:1310 a C-terminal epitope tag 1287:1310 Although overexpression of N-terminal tagged PEBP4 resulted in an inhibition of ERK activation by EGF, that with a C-terminal epitope tag did not have such an effect. 27033522 4 52 theme C 653:653 arg1 terminus 656:663 carboxyl (C) terminus 643:663 carboxyl (C) terminus 643:663 To test if PEBP4 is secreted, we made constructs with Myc epitope at the amino (N) terminus or carboxyl (C) terminus to mask the signal sequence or keep it free, respectively. 21712440 4 0 theme APP/Aβ 868:873 arg1 glycopeptides 875:887 37 APP/Aβ glycopeptides 865:887 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13) 865:965 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 4 1 theme glycopeptides 994:1006 arg1 series 973:978 a series 971:978 a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence 971:1082 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 1 2 theme Aβ 266:267 arg1 -peptides 269:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 9 3 theme sialylated 1687:1696 arg1 O-glycans 1698:1706 sialylated O-glycans 1687:1706 sialylated O-glycans 1687:1706 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 7 4 gly glycosylation 1498:1510 arg1 isoforms 1532:1539 the Aβ1-38/40/42 isoforms 1515:1539 the Aβ1-38/40/42 isoforms 1515:1539 We could not detect any glycosylation of the Aβ1-38/40/42 isoforms. 21712440 3 5 theme chromatography-tandem 665:685 arg1 spectrometry 692:703 then liquid chromatography-tandem mass spectrometry 653:703 then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation 653:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 2 6 contain carrying 424:431 arg2 O-glycans 445:453 O-glycans 445:453 O-glycans 445:453 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 6 contain carrying 424:431 arg1 glycoprotein 411:422 a well-known membrane glycoprotein 389:422 a well-known membrane glycoprotein carrying both N- and O-glycans 389:453 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 6 contain carrying 424:431 arg1 APP 382:384 APP 382:384 APP 382:384 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 6 contain carrying 424:431 arg2 N- 438:439 N- 438:439 N- 438:439 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 1 7 theme amyloid 198:204 arg1 APP 225:227 APP 225:227 APP 225:227 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 1 7 theme amyloid 198:204 arg1 protein 216:222 human amyloid precursor protein 192:222 human amyloid precursor protein (APP) 192:228 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 1 8 theme protein 216:222 arg1 processing 178:187 The proteolytic processing 162:187 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, 162:292 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 2 9 theme glycopeptides 513:525 arg1 occurrence 483:492 the occurrence 479:492 the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF) 479:554 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 3 10 theme structural 709:718 arg1 analysis 720:727 structural analysis 709:727 structural analysis using collision-induced dissociation and electron capture dissociation 709:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 0 11 theme human 135:139 arg1 fluid 155:159 human cerebrospinal fluid 135:159 human cerebrospinal fluid 135:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 2 12 theme released 497:504 arg1 glycopeptides 513:525 released APP/Aβ glycopeptides 497:525 released APP/Aβ glycopeptides 497:525 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 3 13 theme electron 770:777 arg1 dissociation 787:798 electron capture dissociation 770:798 electron capture dissociation 770:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 4 14 gly unglycosylated 819:832 arg1 peptides 841:848 33 unglycosylated APP/Aβ peptides 816:848 33 unglycosylated APP/Aβ peptides 816:848 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 6 15 theme O-acetylated 1398:1409 arg1 terminals 1375:1383 the disialylated terminals 1358:1383 the disialylated terminals occasionally O-acetylated or lactonized 1358:1423 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 4 16 theme Aβ 1072:1073 arg1 sequence 1075:1082 the Aβ sequence 1068:1082 the Aβ sequence 1068:1082 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 8 17 from CSF 1622:1624 arg1 times 1579:1583 up to 2.5 times 1569:1583 up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF 1569:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 4 18 gly glycopeptides 994:1006 arg2 glycopeptides 994:1006 APP/AβX-15 glycopeptides 983:1006 APP/AβX-15 glycopeptides 983:1006 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 6 19 link linked 1286:1291 arg1 structures 1342:1351 (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures 1308:1351 (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures 1308:1351 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 6 19 link linked 1286:1291 arg1 O-glycans 1293:1301 The Tyr10 linked O-glycans 1276:1301 The Tyr10 linked O-glycans 1276:1301 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 0 20 from beta-peptides 118:130 arg1 fluid 155:159 human cerebrospinal fluid 135:159 human cerebrospinal fluid 135:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 8 21 from peptides 1610:1617 arg1 CSF 1622:1624 CSF 1622:1624 CSF 1622:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 5 22 gly glycosylated 1253:1264 arg2 Tyr10 1269:1273 Tyr10 1269:1273 Tyr10 1269:1273 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 9 23 theme mammalian 1757:1765 arg1 protein 1767:1773 a mammalian protein 1755:1773 a mammalian protein 1755:1773 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 6 24 dep HexNAc-O- 1332:1340 arg1 Neu5Ac 1309:1314 Neu5Ac 1309:1314 Neu5Ac 1309:1314 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 0 25 theme tyrosine 57:64 arg1 glycosylations 66:79 threonine, serine, and tyrosine glycosylations 34:79 threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 34:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 1 26 theme critical 310:317 arg1 step 319:322 a critical step 308:322 a critical step in the pathogenesis of Alzheimer's disease (AD) 308:370 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 8 27 theme glycosylated 1594:1605 arg1 peptides 1610:1617 Tyr10 glycosylated Aβ peptides 1588:1617 Tyr10 glycosylated Aβ peptides in CSF 1588:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 2 28 gly glycopeptides 513:525 arg2 glycopeptides 513:525 released APP/Aβ glycopeptides 497:525 released APP/Aβ glycopeptides 497:525 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 7 29 theme isoforms 1532:1539 arg1 glycosylation 1498:1510 any glycosylation 1494:1510 any glycosylation of the Aβ1-38/40/42 isoforms 1494:1539 We could not detect any glycosylation of the Aβ1-38/40/42 isoforms. 21712440 1 30 dep Aβ1-42 286:291 arg1 e.g. 280:283 e.g. 280:283 e.g. 280:283 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 8 31 theme Aβ 1607:1608 arg1 peptides 1610:1617 Tyr10 glycosylated Aβ peptides 1588:1617 Tyr10 glycosylated Aβ peptides in CSF 1588:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 8 32 theme peptides 1610:1617 arg1 times 1579:1583 up to 2.5 times 1569:1583 up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF 1569:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 5 33 theme Aβ1-X 1152:1156 arg1 glycopeptides 1133:1145 27 glycopeptides 1130:1145 27 glycopeptides 1130:1145 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 5 33 theme Aβ1-X 1152:1156 arg1 series 1158:1163 the Aβ1-X series 1148:1163 the Aβ1-X series 1148:1163 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 0 34 theme precursor 92:100 arg1 beta-peptides 118:130 amyloid precursor protein/amyloid beta-peptides 84:130 amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 84:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 6 35 theme terminal 1439:1446 arg1 linkage 1465:1471 a terminal Neu5Acα2,8Neu5Ac linkage 1437:1471 a terminal Neu5Acα2,8Neu5Ac linkage 1437:1471 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 0 36 gly glycosylations 66:79 arg1 beta-peptides 118:130 amyloid precursor protein/amyloid beta-peptides 84:130 amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 84:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 2 37 theme cerebrospinal 530:542 arg1 CSF 551:553 CSF 551:553 CSF 551:553 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 37 theme cerebrospinal 530:542 arg1 fluid 544:548 cerebrospinal fluid 530:548 cerebrospinal fluid (CSF) 530:554 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 0 38 theme beta-peptides 118:130 arg1 glycosylations 66:79 threonine, serine, and tyrosine glycosylations 34:79 threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 34:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 0 39 theme Site-specific 0:12 arg1 characterization 14:29 Site-specific characterization 0:29 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 0:160 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 6 40 theme lactonized 1414:1423 arg1 terminals 1375:1383 the disialylated terminals 1358:1383 the disialylated terminals occasionally O-acetylated or lactonized 1358:1423 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 2 41 theme well-known 391:400 arg1 glycoprotein 411:422 a well-known membrane glycoprotein 389:422 a well-known membrane glycoprotein carrying both N- and O-glycans 389:453 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 41 theme well-known 391:400 arg1 APP 382:384 APP 382:384 APP 382:384 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 9 42 theme amyloidogenic 1820:1832 arg1 pathway 1834:1840 the amyloidogenic pathway 1816:1840 the amyloidogenic pathway associated with AD 1816:1859 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 4 43 theme APP/Aβ 834:839 arg1 peptides 841:848 33 unglycosylated APP/Aβ peptides 816:848 33 unglycosylated APP/Aβ peptides 816:848 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 1 44 from step 319:322 arg1 pathogenesis 331:342 the pathogenesis 327:342 the pathogenesis of Alzheimer's disease (AD) 327:370 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 6 45 dep linked 1286:1291 arg1 Tyr10 1280:1284 Tyr10 1280:1284 Tyr10 1280:1284 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 5 46 gly glycopeptides 1133:1145 arg2 glycopeptides 1133:1145 27 glycopeptides 1130:1145 27 glycopeptides 1130:1145 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 5 46 gly glycopeptides 1133:1145 arg2 series 1158:1163 the Aβ1-X series 1148:1163 the Aβ1-X series 1148:1163 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 8 47 from increase 1557:1564 arg1 patients 1636:1643 six AD patients 1629:1643 six AD patients 1629:1643 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 1 48 theme shorter 235:241 arg1 -peptides 269:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 2 49 gly glycoprotein 411:422 arg1 glycoprotein 411:422 a well-known membrane glycoprotein 389:422 a well-known membrane glycoprotein carrying both N- and O-glycans 389:453 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 49 gly glycoprotein 411:422 arg1 APP 382:384 APP 382:384 APP 382:384 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 1 50 theme proteolytic 166:176 arg1 processing 178:187 The proteolytic processing 162:187 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, 162:292 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 0 51 from fluid 155:159 arg1 glycosylations 66:79 threonine, serine, and tyrosine glycosylations 34:79 threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 34:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 4 52 dep Thr 939:941 arg1 -20 953:955 -20 953:955 -20 953:955 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 4 52 dep Thr 939:941 arg1 -13 962:964 -13 962:964 -13 962:964 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 1 53 theme amyloid 255:261 arg1 -peptides 269:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 8 54 dep 2.5 1575:1577 arg1 to 1572:1573 to 1572:1573 to 1572:1573 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 3 55 theme liquid 658:663 arg1 spectrometry 692:703 then liquid chromatography-tandem mass spectrometry 653:703 then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation 653:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 8 56 theme non-AD 1663:1668 arg1 patients 1670:1677 seven non-AD patients 1657:1677 seven non-AD patients 1657:1677 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 8 57 theme Tyr10 1588:1592 arg1 peptides 1610:1617 Tyr10 glycosylated Aβ peptides 1588:1617 Tyr10 glycosylated Aβ peptides in CSF 1588:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 6 58 gly disialylated 1362:1373 arg1 terminals 1375:1383 the disialylated terminals 1358:1383 the disialylated terminals occasionally O-acetylated or lactonized 1358:1423 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 4 59 gly glycopeptides 875:887 arg2 glycopeptides 875:887 37 APP/Aβ glycopeptides 865:887 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13) 865:965 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 1 60 theme human 192:196 arg1 APP 225:227 APP 225:227 APP 225:227 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 1 60 theme human 192:196 arg1 protein 216:222 human amyloid precursor protein 192:222 human amyloid precursor protein (APP) 192:228 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 3 61 theme mass 687:690 arg1 spectrometry 692:703 then liquid chromatography-tandem mass spectrometry 653:703 then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation 653:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 9 62 gly sialylated 1687:1696 arg1 O-glycans 1698:1706 sialylated O-glycans 1687:1706 sialylated O-glycans 1687:1706 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 1 63 theme precursor 206:214 arg1 APP 225:227 APP 225:227 APP 225:227 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 1 63 theme precursor 206:214 arg1 protein 216:222 human amyloid precursor protein 192:222 human amyloid precursor protein (APP) 192:228 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 3 64 theme collision-induced 735:751 arg1 dissociation 753:764 collision-induced dissociation 735:764 collision-induced dissociation 735:764 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 0 65 theme cerebrospinal 141:153 arg1 fluid 155:159 human cerebrospinal fluid 135:159 human cerebrospinal fluid 135:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 2 66 theme APP/Aβ 506:511 arg1 glycopeptides 513:525 released APP/Aβ glycopeptides 497:525 released APP/Aβ glycopeptides 497:525 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 9 67 theme Tyr 1729:1731 arg1 first 1746:1750 first 1746:1750 first 1746:1750 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 9 67 theme Tyr 1729:1731 arg1 residue 1733:1739 a Tyr residue 1727:1739 a Tyr residue 1727:1739 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 3 68 from samples 641:647 arg1 glycopeptides 618:630 glycopeptides 618:630 glycopeptides 618:630 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 3 68 from samples 641:647 arg1 peptides 605:612 Aβ peptides 602:612 Aβ peptides 602:612 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 3 68 from samples 641:647 arg1 spectrometry 692:703 then liquid chromatography-tandem mass spectrometry 653:703 then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation 653:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 0 69 from glycosylations 66:79 arg1 fluid 155:159 human cerebrospinal fluid 135:159 human cerebrospinal fluid 135:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 4 70 theme sequence 1075:1082 arg1 Asp1 1060:1063 Asp1 1060:1063 Asp1 of the Aβ sequence 1060:1082 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 8 71 from times 1579:1583 arg1 CSF 1622:1624 CSF 1622:1624 CSF 1622:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 6 72 with structures 1342:1351 arg1 terminals 1375:1383 the disialylated terminals 1358:1383 the disialylated terminals occasionally O-acetylated or lactonized 1358:1423 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 4 73 theme sialylated 894:903 arg1 core 905:908 sialylated core 1 894:910 sialylated core 1 894:910 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 3 74 theme CSF 637:639 arg1 samples 641:647 CSF samples 637:647 CSF samples 637:647 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 3 75 theme 6E10 569:572 arg1 antibody 574:581 the 6E10 antibody 565:581 the 6E10 antibody 565:581 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 6 76 theme disialylated 1362:1373 arg1 terminals 1375:1383 the disialylated terminals 1358:1383 the disialylated terminals occasionally O-acetylated or lactonized 1358:1423 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 4 77 from -63 1021:1023 arg1 relation 1048:1055 relation 1048:1055 relation to Asp1 of the Aβ sequence 1048:1082 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 4 78 with glycopeptides 875:887 arg1 core 905:908 sialylated core 1 894:910 sialylated core 1 894:910 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 2 79 from occurrence 483:492 arg1 CSF 551:553 CSF 551:553 CSF 551:553 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 79 from occurrence 483:492 arg1 fluid 544:548 cerebrospinal fluid 530:548 cerebrospinal fluid (CSF) 530:554 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 0 80 theme glycosylations 66:79 arg1 characterization 14:29 Site-specific characterization 0:29 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 0:160 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 8 81 theme times 1579:1583 arg1 increase 1557:1564 an increase 1554:1564 an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients 1554:1643 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 4 82 gly sialylated 894:903 arg1 core 905:908 sialylated core 1 894:910 sialylated core 1 894:910 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 3 83 used used 560:563 arg2 We 557:558 We 557:558 We 557:558 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 5 84 theme glycopeptides 1133:1145 arg1 series 1120:1125 a series 1118:1125 a series 1118:1125 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 5 84 theme glycopeptides 1133:1145 arg1 all 1240:1242 all 1240:1242 all 1240:1242 Unexpectedly, we also identified a series of 27 glycopeptides, the Aβ1-X series, where X was 20 (DAEFRHDSGYEVHHQKLVFF), 19, 18, 17, 16, and 15, which were all uniquely glycosylated on Tyr10. 21712440 0 85 theme amyloid 84:90 arg1 beta-peptides 118:130 amyloid precursor protein/amyloid beta-peptides 84:130 amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 84:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 6 86 theme HexNAc-O- 1332:1340 arg1 structures 1342:1351 (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures 1308:1351 (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures 1308:1351 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 6 86 theme HexNAc-O- 1332:1340 arg1 O-glycans 1293:1301 The Tyr10 linked O-glycans 1276:1301 The Tyr10 linked O-glycans 1276:1301 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 0 87 theme protein/amyloid 102:116 arg1 beta-peptides 118:130 amyloid precursor protein/amyloid beta-peptides 84:130 amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 84:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 6 88 theme Neu5Acα2,8Neu5Ac 1448:1463 arg1 linkage 1465:1471 a terminal Neu5Acα2,8Neu5Ac linkage 1437:1471 a terminal Neu5Acα2,8Neu5Ac linkage 1437:1471 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 9 89 theme APP 1789:1791 arg1 processing 1793:1802 APP processing 1789:1802 APP processing 1789:1802 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 4 90 attach attached 927:934 arg2 O-glycans 917:925 O-glycans 917:925 O-glycans attached to Thr(-39, -21, -20, and -13) 917:965 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 4 90 attach attached 927:934 arg1 Thr 939:941 Thr 939:941 Thr(-39, -21, -20, and -13) 939:965 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 9 91 from protein 1767:1773 arg1 first 1746:1750 first 1746:1750 first 1746:1750 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 9 91 from protein 1767:1773 arg1 residue 1733:1739 a Tyr residue 1727:1739 a Tyr residue 1727:1739 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 8 92 gly glycosylated 1594:1605 arg1 peptides 1610:1617 Tyr10 glycosylated Aβ peptides 1588:1617 Tyr10 glycosylated Aβ peptides in CSF 1588:1624 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 2 93 theme membrane 402:409 arg1 glycoprotein 411:422 a well-known membrane glycoprotein 389:422 a well-known membrane glycoprotein carrying both N- and O-glycans 389:453 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 2 93 theme membrane 402:409 arg1 APP 382:384 APP 382:384 APP 382:384 Although APP is a well-known membrane glycoprotein carrying both N- and O-glycans, nothing is known about the occurrence of released APP/Aβ glycopeptides in cerebrospinal fluid (CSF). 21712440 0 94 theme threonine 34:42 arg1 glycosylations 66:79 threonine, serine, and tyrosine glycosylations 34:79 threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 34:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 8 95 theme AD 1633:1634 arg1 patients 1636:1643 six AD patients 1629:1643 six AD patients 1629:1643 We observed an increase of up to 2.5 times of Tyr10 glycosylated Aβ peptides in CSF in six AD patients compared to seven non-AD patients. 21712440 9 96 from first 1746:1750 arg1 protein 1767:1773 a mammalian protein 1755:1773 a mammalian protein 1755:1773 APP/Aβ sialylated O-glycans, including that of a Tyr residue, the first in a mammalian protein, may modulate APP processing, inhibiting the amyloidogenic pathway associated with AD. 21712440 3 97 theme Aβ 602:603 arg1 peptides 605:612 Aβ peptides 602:612 Aβ peptides 602:612 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 0 98 theme serine 45:50 arg1 glycosylations 66:79 threonine, serine, and tyrosine glycosylations 34:79 threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid 34:159 Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid beta-peptides in human cerebrospinal fluid. 21712440 4 99 theme unglycosylated 819:832 arg1 peptides 841:848 33 unglycosylated APP/Aβ peptides 816:848 33 unglycosylated APP/Aβ peptides 816:848 In addition to 33 unglycosylated APP/Aβ peptides, we identified 37 APP/Aβ glycopeptides with sialylated core 1 like O-glycans attached to Thr(-39, -21, -20, and -13), in a series of APP/AβX-15 glycopeptides, where X was -63, -57, -52, and -45, in relation to Asp1 of the Aβ sequence. 21712440 1 100 theme aggregating 243:253 arg1 -peptides 269:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 3 101 gly glycopeptides 618:630 arg2 glycopeptides 618:630 glycopeptides 618:630 glycopeptides 618:630 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 3 101 gly glycopeptides 618:630 arg1 samples 641:647 CSF samples 637:647 CSF samples 637:647 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 21712440 1 102 theme β 263:263 arg1 -peptides 269:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 shorter aggregating amyloid β (Aβ)-peptides 235:277 The proteolytic processing of human amyloid precursor protein (APP) into shorter aggregating amyloid β (Aβ)-peptides, e.g., Aβ1-42, is considered a critical step in the pathogenesis of Alzheimer's disease (AD). 21712440 6 103 theme linked 1286:1291 arg1 structures 1342:1351 (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures 1308:1351 (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures 1308:1351 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 6 103 theme linked 1286:1291 arg1 O-glycans 1293:1301 The Tyr10 linked O-glycans 1276:1301 The Tyr10 linked O-glycans 1276:1301 The Tyr10 linked O-glycans were (Neu5Ac)(1-2)Hex(Neu5Ac)HexNAc-O- structures with the disialylated terminals occasionally O-acetylated or lactonized, indicating a terminal Neu5Acα2,8Neu5Ac linkage. 21712440 3 104 theme capture 779:785 arg1 dissociation 787:798 electron capture dissociation 770:798 electron capture dissociation 770:798 We used the 6E10 antibody and immunopurified Aβ peptides and glycopeptides from CSF samples and then liquid chromatography-tandem mass spectrometry for structural analysis using collision-induced dissociation and electron capture dissociation. 9030779 0 0 theme site-directed 91:103 arg1 mutagenesis 105:115 site-directed mutagenesis 91:115 site-directed mutagenesis 91:115 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 5 1 theme lysosomal 706:714 arg1 targeting 716:724 lysosomal targeting 706:724 lysosomal targeting 706:724 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 1 2 theme marker 191:196 arg1 present 198:204 a mannose-6-phosphate recognition marker present 157:204 a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes 157:276 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 7 3 theme primary 933:939 arg1 product 953:959 the primary translation product 929:959 the primary translation product 929:959 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 7 4 theme glycosylation 1076:1088 arg1 site 1090:1093 glycosylation site six 1076:1097 glycosylation site six 1076:1097 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 1 5 from present 198:204 arg1 oligosaccharides 227:242 asparagine-linked oligosaccharides 209:242 asparagine-linked oligosaccharides 209:242 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 2 6 theme oligosaccharide 342:356 arg1 chains 358:363 the six potential N-linked oligosaccharide chains 315:363 the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) 315:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 1 7 theme soluble 123:129 arg1 enzymes 141:147 Most soluble lysosomal enzymes 118:147 Most soluble lysosomal enzymes 118:147 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 6 8 gly N-glycosylation 801:815 arg2 sites 817:821 the two C-terminal N-glycosylation sites 782:821 the two C-terminal N-glycosylation sites 782:821 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 6 8 gly N-glycosylation 801:815 arg2 two 786:788 two 786:788 two 786:788 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 7 9 gly glycosylation 875:887 arg2 site 889:892 glycosylation site five 875:897 glycosylation site five 875:897 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 7 9 gly glycosylation 875:887 arg2 five 894:897 five 894:897 five 894:897 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 5 10 theme N-terminal 658:667 arg1 sites 683:687 the four N-terminal glycosylation sites 649:687 the four N-terminal glycosylation sites 649:687 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 2 11 theme N-linked 333:340 arg1 chains 358:363 the six potential N-linked oligosaccharide chains 315:363 the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) 315:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 8 12 theme cleavage 1273:1280 arg1 site 1247:1250 the site 1243:1250 the site of early proteolytic cleavage of newly synthesized ASM 1243:1305 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 8 12 theme cleavage 1273:1280 arg1 located 1315:1321 located 1315:1321 located 1315:1321 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 8 12 theme cleavage 1273:1280 arg1 cleavage 1273:1280 early proteolytic cleavage 1255:1280 early proteolytic cleavage of newly synthesized ASM 1255:1305 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 8 13 theme early 1255:1259 arg1 cleavage 1273:1280 early proteolytic cleavage 1255:1280 early proteolytic cleavage of newly synthesized ASM 1255:1305 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 5 14 theme sites 683:687 arg1 Elimination 634:644 Elimination 634:644 Elimination of the four N-terminal glycosylation sites 634:687 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 5 15 theme glycosylation 669:681 arg1 sites 683:687 the four N-terminal glycosylation sites 649:687 the four N-terminal glycosylation sites 649:687 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 2 16 theme potential 323:331 arg1 chains 358:363 the six potential N-linked oligosaccharide chains 315:363 the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) 315:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 7 17 dep led 1099:1101 arg1 retained 1155:1162 retained 1155:1162 retained inside the endoplasmic reticulum/Golgi 1155:1201 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 7 18 theme endoplasmic 1175:1185 arg1 reticulum/Golgi 1187:1201 the endoplasmic reticulum/Golgi 1171:1201 the endoplasmic reticulum/Golgi 1171:1201 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 6 19 theme enzyme 856:861 arg1 formation 836:844 the formation 832:844 the formation of mature enzyme 832:861 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 8 20 theme proteolytic 1261:1271 arg1 cleavage 1273:1280 early proteolytic cleavage 1255:1280 early proteolytic cleavage of newly synthesized ASM 1255:1305 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 7 21 theme rapid 911:915 arg1 cleavage 917:924 rapid cleavage 911:924 rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi 911:1201 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 1 22 theme asparagine-linked 209:225 arg1 oligosaccharides 227:242 asparagine-linked oligosaccharides 209:242 asparagine-linked oligosaccharides 209:242 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 7 23 theme endoplasmic 1027:1037 arg1 reticulum/Golgi 1039:1053 the endoplasmic reticulum/Golgi 1023:1053 the endoplasmic reticulum/Golgi 1023:1053 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 4 24 theme sites 618:622 arg1 five 604:607 five 604:607 five 604:607 Evidence is presented that five of these sites are used. 9030779 4 24 theme sites 618:622 arg1 sites 618:622 these sites 612:622 these sites 612:622 Evidence is presented that five of these sites are used. 9030779 6 25 theme mature 849:854 arg1 enzyme 856:861 mature enzyme 849:861 mature enzyme 849:861 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 2 26 theme acid 374:377 arg1 ASM 397:399 ASM 397:399 ASM 397:399 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 26 theme acid 374:377 arg1 sphingomyelinase 379:394 human acid sphingomyelinase 368:394 human acid sphingomyelinase (ASM) 368:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 8 27 theme second 1335:1340 arg1 sites 1366:1370 the second and third glycosylation sites 1331:1370 the second and third glycosylation sites 1331:1370 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 0 28 theme Functional 0:9 arg1 characterization 11:26 Functional characterization 0:26 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 0:116 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 1 29 link asparagine-linked 209:225 arg1 oligosaccharides 227:242 asparagine-linked oligosaccharides 209:242 asparagine-linked oligosaccharides 209:242 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 2 30 theme human 368:372 arg1 ASM 397:399 ASM 397:399 ASM 397:399 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 30 theme human 368:372 arg1 sphingomyelinase 379:394 human acid sphingomyelinase 368:394 human acid sphingomyelinase (ASM) 368:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 31 from influence 302:310 arg1 targeting 425:433 targeting 425:433 targeting 425:433 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 31 from influence 302:310 arg1 processing 440:449 processing 440:449 processing 440:449 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 31 from influence 302:310 arg1 activity 415:422 catalytic activity 405:422 catalytic activity 405:422 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 3 32 theme N-glycosylation 471:485 arg1 site 487:490 Each N-glycosylation site 466:490 Each N-glycosylation site 466:490 Each N-glycosylation site was modified by site-directed mutagenesis and subsequently expressed in COS-1 cells. 9030779 7 33 gly glycosylation 1076:1088 arg2 site 1090:1093 glycosylation site six 1076:1097 glycosylation site six 1076:1097 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 7 33 gly glycosylation 1076:1088 arg2 six 1095:1097 six 1095:1097 six 1095:1097 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 1 34 theme Most 118:121 arg1 enzymes 141:147 Most soluble lysosomal enzymes 118:147 Most soluble lysosomal enzymes 118:147 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 1 35 theme proper 248:253 arg1 targeting 255:263 proper targeting 248:263 proper targeting to lysosomes 248:276 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 7 36 theme site 1090:1093 arg1 deletion 1064:1071 deletion 1064:1071 deletion of glycosylation site six 1064:1097 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 8 37 theme third 1346:1350 arg1 sites 1366:1370 the second and third glycosylation sites 1331:1370 the second and third glycosylation sites 1331:1370 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 2 38 theme chains 358:363 arg1 influence 302:310 the influence 298:310 the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme 298:463 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 7 39 theme site 889:892 arg1 Absence 864:870 Absence 864:870 Absence of glycosylation site five 864:897 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 2 40 theme enzyme 458:463 arg1 targeting 425:433 targeting 425:433 targeting 425:433 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 40 theme enzyme 458:463 arg1 processing 440:449 processing 440:449 processing 440:449 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 2 40 theme enzyme 458:463 arg1 activity 415:422 catalytic activity 405:422 catalytic activity 405:422 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 0 41 theme sites 51:55 arg1 characterization 11:26 Functional characterization 0:26 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 0:116 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 2 42 link N-linked 333:340 arg1 chains 358:363 the six potential N-linked oligosaccharide chains 315:363 the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) 315:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 7 43 theme ASM 1135:1137 arg1 precursor 1139:1147 an inactive ASM precursor 1123:1147 an inactive ASM precursor 1123:1147 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 0 44 theme N-glycosylation 35:49 arg1 sites 51:55 the N-glycosylation sites 31:55 the N-glycosylation sites of human acid sphingomyelinase 31:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 0 44 theme N-glycosylation 35:49 arg1 sphingomyelinase 71:86 human acid sphingomyelinase 60:86 human acid sphingomyelinase 60:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 2 45 theme catalytic 405:413 arg1 activity 415:422 catalytic activity 405:422 catalytic activity 405:422 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 7 46 theme precursor 1139:1147 arg1 formation 1110:1118 the formation 1106:1118 the formation of an inactive ASM precursor 1106:1147 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 8 47 theme glycosylation 1352:1364 arg1 sites 1366:1370 the second and third glycosylation sites 1331:1370 the second and third glycosylation sites 1331:1370 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 0 48 theme human 60:64 arg1 sphingomyelinase 71:86 human acid sphingomyelinase 60:86 human acid sphingomyelinase 60:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 3 49 theme COS-1 564:568 arg1 cells 570:574 COS-1 cells 564:574 COS-1 cells 564:574 Each N-glycosylation site was modified by site-directed mutagenesis and subsequently expressed in COS-1 cells. 9030779 8 50 theme ASM 1303:1305 arg1 cleavage 1273:1280 early proteolytic cleavage 1255:1280 early proteolytic cleavage of newly synthesized ASM 1255:1305 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 7 51 theme glycosylation 875:887 arg1 site 889:892 glycosylation site five 875:897 glycosylation site five 875:897 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 5 52 theme enzymatic 742:750 arg1 activity 752:759 enzymatic activity 742:759 enzymatic activity 742:759 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 1 53 theme lysosomal 131:139 arg1 enzymes 141:147 Most soluble lysosomal enzymes 118:147 Most soluble lysosomal enzymes 118:147 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 7 54 theme inactive 1126:1133 arg1 precursor 1139:1147 an inactive ASM precursor 1123:1147 an inactive ASM precursor 1123:1147 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 6 55 theme sites 817:821 arg1 removal 771:777 removal 771:777 removal of the two C-terminal N-glycosylation sites 771:821 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 3 56 mod modified 496:503 arg1 site 487:490 Each N-glycosylation site 466:490 Each N-glycosylation site 466:490 Each N-glycosylation site was modified by site-directed mutagenesis and subsequently expressed in COS-1 cells. 9030779 3 56 mod modified 496:503 arg3 mutagenesis 522:532 site-directed mutagenesis 508:532 site-directed mutagenesis 508:532 Each N-glycosylation site was modified by site-directed mutagenesis and subsequently expressed in COS-1 cells. 9030779 7 57 theme inactive 981:988 arg1 protein 990:996 an enzymatically inactive protein 964:996 an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi 964:1201 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 4 58 used used 628:631 arg2 sites 618:622 these sites 612:622 these sites 612:622 Evidence is presented that five of these sites are used. 9030779 4 58 used used 628:631 arg2 five 604:607 five 604:607 five 604:607 Evidence is presented that five of these sites are used. 9030779 0 59 gly N-glycosylation 35:49 arg2 sites 51:55 the N-glycosylation sites 31:55 the N-glycosylation sites of human acid sphingomyelinase 31:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 0 59 gly N-glycosylation 35:49 arg1 sphingomyelinase 71:86 human acid sphingomyelinase 60:86 human acid sphingomyelinase 60:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 0 59 gly N-glycosylation 35:49 arg2 sphingomyelinase 71:86 human acid sphingomyelinase 60:86 human acid sphingomyelinase 60:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 6 60 theme N-glycosylation 801:815 arg1 sites 817:821 the two C-terminal N-glycosylation sites 782:821 the two C-terminal N-glycosylation sites 782:821 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 8 61 gly glycosylation 1352:1364 arg2 sites 1366:1370 the second and third glycosylation sites 1331:1370 the second and third glycosylation sites 1331:1370 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 6 62 theme C-terminal 790:799 arg1 sites 817:821 the two C-terminal N-glycosylation sites 782:821 the two C-terminal N-glycosylation sites 782:821 However, removal of the two C-terminal N-glycosylation sites inhibits the formation of mature enzyme. 9030779 2 63 theme sphingomyelinase 379:394 arg1 chains 358:363 the six potential N-linked oligosaccharide chains 315:363 the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) 315:400 We have determined the influence of the six potential N-linked oligosaccharide chains of human acid sphingomyelinase (ASM) on catalytic activity, targeting, and processing of the enzyme. 9030779 8 64 theme synthesized 1291:1301 arg1 ASM 1303:1305 newly synthesized ASM 1285:1305 newly synthesized ASM 1285:1305 Our results also provide evidence that the site of early proteolytic cleavage of newly synthesized ASM must be located between the second and third glycosylation sites. 9030779 0 65 theme sphingomyelinase 71:86 arg1 sites 51:55 the N-glycosylation sites 31:55 the N-glycosylation sites of human acid sphingomyelinase 31:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 0 65 theme sphingomyelinase 71:86 arg1 sphingomyelinase 71:86 human acid sphingomyelinase 60:86 human acid sphingomyelinase 60:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 7 66 theme translation 941:951 arg1 product 953:959 the primary translation product 929:959 the primary translation product 929:959 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 9030779 3 67 gly N-glycosylation 471:485 arg2 site 487:490 Each N-glycosylation site 466:490 Each N-glycosylation site 466:490 Each N-glycosylation site was modified by site-directed mutagenesis and subsequently expressed in COS-1 cells. 9030779 3 68 theme site-directed 508:520 arg1 mutagenesis 522:532 site-directed mutagenesis 508:532 site-directed mutagenesis 508:532 Each N-glycosylation site was modified by site-directed mutagenesis and subsequently expressed in COS-1 cells. 9030779 1 69 theme mannose-6-phosphate 159:177 arg1 present 198:204 a mannose-6-phosphate recognition marker present 157:204 a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes 157:276 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 5 70 gly glycosylation 669:681 arg2 sites 683:687 the four N-terminal glycosylation sites 649:687 the four N-terminal glycosylation sites 649:687 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 5 70 gly glycosylation 669:681 arg2 four 653:656 four 653:656 four 653:656 Elimination of the four N-terminal glycosylation sites does not disturb lysosomal targeting, processing, or enzymatic activity. 9030779 0 71 theme acid 66:69 arg1 sphingomyelinase 71:86 human acid sphingomyelinase 60:86 human acid sphingomyelinase 60:86 Functional characterization of the N-glycosylation sites of human acid sphingomyelinase by site-directed mutagenesis. 9030779 1 72 theme recognition 179:189 arg1 present 198:204 a mannose-6-phosphate recognition marker present 157:204 a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes 157:276 Most soluble lysosomal enzymes require a mannose-6-phosphate recognition marker present on asparagine-linked oligosaccharides for proper targeting to lysosomes. 9030779 7 73 theme product 953:959 arg1 cleavage 917:924 rapid cleavage 911:924 rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi 911:1201 Absence of glycosylation site five resulted in rapid cleavage of the primary translation product to an enzymatically inactive protein which accumulated inside the endoplasmic reticulum/Golgi, whereas deletion of glycosylation site six led to the formation of an inactive ASM precursor, also retained inside the endoplasmic reticulum/Golgi. 12970363 4 0 theme fluorescence 601:612 arg1 imaging 614:620 fluorescence imaging 601:620 fluorescence imaging 601:620 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 6 1 from motifs 906:911 arg1 TRPC6 916:920 TRPC6 916:920 TRPC6 916:920 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 6 2 link N-linked 1008:1015 arg1 sites 1031:1035 both extracellular N-linked glycosylation sites 989:1035 both extracellular N-linked glycosylation sites 989:1035 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 1 3 theme functional 246:255 arg1 similarities 257:268 functional similarities 246:268 sequence homology as well as functional similarities 217:268 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 0 4 link N-linked 0:7 arg1 glycosylation 17:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 0 4 link N-linked 0:7 arg1 determinant 42:52 a major determinant 34:52 a major determinant for basal TRPC3 and TRPC6 channel activity 34:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 10 5 gly glycosylation 1397:1409 arg2 site 1411:1414 the e2 glycosylation site 1390:1414 the e2 glycosylation site 1390:1414 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 7 6 theme wild 1103:1106 arg1 type 1108:1111 TRPC6 wild type 1097:1111 TRPC6 wild type 1097:1111 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 2 7 theme subfamily 296:304 arg1 Members 271:277 Members 271:277 Members of the TRPC3/6/7 subfamily 271:304 Members of the TRPC3/6/7 subfamily share common biophysical characteristics and are activated by diacylglycerol in a membrane-delimited manner. 12970363 11 8 from TRPC3 1676:1680 arg1 engineering 1619:1629 engineering 1619:1629 engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 1619:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 4 9 theme considerable 661:672 arg1 activity 687:694 considerable constitutive activity 661:694 considerable constitutive activity 661:694 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 11 10 theme glycosylation 1695:1707 arg1 status 1709:1714 the glycosylation status 1691:1714 the glycosylation status in TRPC6 1691:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 12 11 theme TRPC6 1856:1860 arg1 regulation 1842:1851 the tight regulation 1832:1851 the tight regulation of TRPC6 through phospholipase C-activating receptors 1832:1905 We conclude that the glycosylation pattern plays a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors. 12970363 11 12 theme TRPC3 1742:1746 arg1 activity 1754:1761 TRPC3 basal activity 1742:1761 TRPC3 basal activity 1742:1761 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 7 13 theme cell 1073:1076 arg1 lysates 1078:1084 HEK 293 cell lysates 1065:1084 HEK 293 cell lysates expressing TRPC6 wild type and mutants 1065:1123 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 12 14 theme phospholipase 1870:1882 arg1 receptors 1897:1905 phospholipase C-activating receptors 1870:1905 phospholipase C-activating receptors 1870:1905 We conclude that the glycosylation pattern plays a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors. 12970363 10 15 theme receptor-regulated 1494:1511 arg1 TRPC6 1513:1517 the tightly receptor-regulated TRPC6 1482:1517 the tightly receptor-regulated TRPC6 1482:1517 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 7 16 theme HEK 1065:1067 arg1 lysates 1078:1084 HEK 293 cell lysates 1065:1084 HEK 293 cell lysates expressing TRPC6 wild type and mutants 1065:1123 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 11 17 from site 1668:1671 arg1 TRPC3 1676:1680 TRPC3 1676:1680 TRPC3 1676:1680 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 11 18 theme site 1668:1671 arg1 engineering 1619:1629 engineering 1619:1629 engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 1619:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 1 19 theme receptor-activated 117:134 arg1 channels 143:150 receptor-activated cation channels 117:150 receptor-activated cation channels (TRPC channels) 117:166 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 1 19 theme receptor-activated 117:134 arg1 channels 158:165 TRPC channels 153:165 TRPC channels 153:165 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 11 20 from engineering 1619:1629 arg1 TRPC3 1676:1680 TRPC3 1676:1680 TRPC3 1676:1680 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 10 21 gly monoglycosylated 1432:1447 arg1 TRPC3 1449:1453 the monoglycosylated TRPC3 1428:1453 the monoglycosylated TRPC3 1428:1453 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 11 22 theme second 1648:1653 arg1 site 1668:1671 an additional second glycosylated site 1634:1671 an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 1634:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 5 23 theme functional 803:812 arg1 difference 814:823 the functional difference 799:823 the functional difference 799:823 To identify potential molecular correlates accounting for the functional difference, we analyzed the glycosylation pattern of TRPC6 compared with TRPC3. 12970363 1 24 theme channels 143:150 arg1 family 107:112 The TRPC family 98:112 The TRPC family of receptor-activated cation channels (TRPC channels) 98:166 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 6 25 theme N-linked 1008:1015 arg1 sites 1031:1035 both extracellular N-linked glycosylation sites 989:1035 both extracellular N-linked glycosylation sites 989:1035 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 7 26 theme second 1203:1208 arg1 e2 1230:1231 e2 1230:1231 e2 1230:1231 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 26 theme second 1203:1208 arg1 loop 1224:1227 the first (e1) and second extracellular loop 1184:1227 the first (e1) and second extracellular loop (e2) 1184:1232 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 1 27 theme TRPC 153:156 arg1 channels 143:150 receptor-activated cation channels 117:150 receptor-activated cation channels (TRPC channels) 117:166 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 1 27 theme TRPC 153:156 arg1 channels 158:165 TRPC channels 153:165 TRPC channels 153:165 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 0 28 theme channel 80:86 arg1 activity 88:95 basal TRPC3 and TRPC6 channel activity 58:95 basal TRPC3 and TRPC6 channel activity 58:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 3 29 theme cellular 557:564 arg1 roles 566:570 distinct cellular roles 548:570 distinct cellular roles 548:570 At present, it is only poorly understood whether members of the TRPC3/6/7 subfamily are functionally redundant or whether they serve distinct cellular roles. 12970363 4 30 theme imaging 614:620 arg1 strategies 622:631 electrophysiological and fluorescence imaging strategies 576:631 electrophysiological and fluorescence imaging strategies 576:631 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 0 31 theme TRPC3 64:68 arg1 activity 88:95 basal TRPC3 and TRPC6 channel activity 58:95 basal TRPC3 and TRPC6 channel activity 58:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 10 32 theme active 1541:1546 arg1 channel 1548:1554 a constitutively active channel 1524:1554 a constitutively active channel 1524:1554 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 0 33 theme N-linked 0:7 arg1 glycosylation 17:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 0 33 theme N-linked 0:7 arg1 determinant 42:52 a major determinant 34:52 a major determinant for basal TRPC3 and TRPC6 channel activity 34:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 12 34 theme pivotal 1815:1821 arg1 role 1823:1826 a pivotal role 1813:1826 a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors 1813:1905 We conclude that the glycosylation pattern plays a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors. 12970363 11 35 gly glycosylated 1655:1666 arg1 site 1668:1671 an additional second glycosylated site 1634:1671 an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 1634:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 10 36 theme functional 1568:1577 arg1 characteristics 1579:1593 functional characteristics 1568:1593 functional characteristics of TRPC3 1568:1602 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 12 37 theme glycosylation 1785:1797 arg1 pattern 1799:1805 the glycosylation pattern 1781:1805 the glycosylation pattern 1781:1805 We conclude that the glycosylation pattern plays a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors. 12970363 7 38 dep first 1188:1192 arg1 e1 1195:1196 e1 1195:1196 e1 1195:1196 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 0 39 theme basal 58:62 arg1 activity 88:95 basal TRPC3 and TRPC6 channel activity 58:95 basal TRPC3 and TRPC6 channel activity 58:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 3 40 theme subfamily 489:497 arg1 members 464:470 members 464:470 members of the TRPC3/6/7 subfamily 464:497 At present, it is only poorly understood whether members of the TRPC3/6/7 subfamily are functionally redundant or whether they serve distinct cellular roles. 12970363 5 41 theme glycosylation 842:854 arg1 pattern 856:862 the glycosylation pattern 838:862 the glycosylation pattern of TRPC6 838:871 To identify potential molecular correlates accounting for the functional difference, we analyzed the glycosylation pattern of TRPC6 compared with TRPC3. 12970363 7 42 dep Zhu 1297:1299 arg1 Biol 1346:1349 Biol 1346:1349 Biol 1346:1349 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 43 theme monoglycosylated 1252:1267 arg1 channel 1275:1281 the monoglycosylated TRPC3 channel 1248:1281 the monoglycosylated TRPC3 channel 1248:1281 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 10 44 theme glycosylation 1397:1409 arg1 site 1411:1414 the e2 glycosylation site 1390:1414 the e2 glycosylation site 1390:1414 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 11 45 theme additional 1637:1646 arg1 site 1668:1671 an additional second glycosylated site 1634:1671 an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 1634:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 2 46 theme biophysical 319:329 arg1 characteristics 331:345 common biophysical characteristics 312:345 common biophysical characteristics 312:345 Members of the TRPC3/6/7 subfamily share common biophysical characteristics and are activated by diacylglycerol in a membrane-delimited manner. 12970363 10 47 theme monoglycosylated 1432:1447 arg1 TRPC3 1449:1453 the monoglycosylated TRPC3 1428:1453 the monoglycosylated TRPC3 1428:1453 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 9 48 dep 273 1358:1360 arg1 8675-8679 1363:1371 8675-8679 1363:1371 8675-8679 1363:1371 273, 8675-8679). 12970363 1 49 theme sequence 217:224 arg1 homology 226:233 sequence homology 217:233 sequence homology as well as functional similarities 217:268 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 6 50 gly glycosylation 1017:1029 arg2 sites 1031:1035 both extracellular N-linked glycosylation sites 989:1035 both extracellular N-linked glycosylation sites 989:1035 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 6 51 dep mutated 927:933 arg1 Asn 936:938 Asn 936:938 Asn to Gln 936:945 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 3 52 theme distinct 548:555 arg1 roles 566:570 distinct cellular roles 548:570 distinct cellular roles 548:570 At present, it is only poorly understood whether members of the TRPC3/6/7 subfamily are functionally redundant or whether they serve distinct cellular roles. 12970363 6 53 dep Asn 936:938 arg1 Gln 943:945 Gln 943:945 Asn to Gln 936:945 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 6 53 dep Asn 936:938 arg1 to 940:941 to 940:941 to 940:941 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 7 54 theme TRPC6 1143:1147 arg1 model 1134:1138 a model 1132:1138 a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol 1132:1349 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 11 55 from status 1709:1714 arg1 TRPC6 1719:1723 TRPC6 1719:1723 TRPC6 1719:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 7 56 theme TRPC6 1097:1101 arg1 type 1108:1111 TRPC6 wild type 1097:1111 TRPC6 wild type 1097:1111 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 12 57 theme C-activating 1884:1895 arg1 receptors 1897:1905 phospholipase C-activating receptors 1870:1905 phospholipase C-activating receptors 1870:1905 We conclude that the glycosylation pattern plays a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors. 12970363 5 58 gly glycosylation 842:854 arg1 TRPC6 867:871 TRPC6 867:871 TRPC6 867:871 To identify potential molecular correlates accounting for the functional difference, we analyzed the glycosylation pattern of TRPC6 compared with TRPC3. 12970363 7 59 theme lysates 1078:1084 arg1 analysis 1053:1060 Immunoblotting analysis 1038:1060 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants 1038:1123 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 6 60 theme in 951:952 arg1 mutagenesis 960:970 in vitro mutagenesis 951:970 in vitro mutagenesis 951:970 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 6 61 dep in 951:952 arg1 vitro 954:958 vitro 954:958 vitro 954:958 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 4 62 theme constitutive 674:685 arg1 activity 687:694 considerable constitutive activity 661:694 considerable constitutive activity 661:694 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 11 63 theme basal 1748:1752 arg1 activity 1754:1761 TRPC3 basal activity 1742:1761 TRPC3 basal activity 1742:1761 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 1 64 theme TRPC 102:105 arg1 family 107:112 The TRPC family 98:112 The TRPC family of receptor-activated cation channels (TRPC channels) 98:166 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 7 65 theme Immunoblotting 1038:1051 arg1 analysis 1053:1060 Immunoblotting analysis 1038:1060 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants 1038:1123 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 66 theme extracellular 1210:1222 arg1 e2 1230:1231 e2 1230:1231 e2 1230:1231 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 66 theme extracellular 1210:1222 arg1 loop 1224:1227 the first (e1) and second extracellular loop 1184:1227 the first (e1) and second extracellular loop (e2) 1184:1232 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 1 67 theme cation 136:141 arg1 channels 143:150 receptor-activated cation channels 117:150 receptor-activated cation channels (TRPC channels) 117:166 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 1 67 theme cation 136:141 arg1 channels 158:165 TRPC channels 153:165 TRPC channels 153:165 The TRPC family of receptor-activated cation channels (TRPC channels) can be subdivided into four subfamilies based on sequence homology as well as functional similarities. 12970363 3 68 theme TRPC3/6/7 479:487 arg1 subfamily 489:497 the TRPC3/6/7 subfamily 475:497 the TRPC3/6/7 subfamily 475:497 At present, it is only poorly understood whether members of the TRPC3/6/7 subfamily are functionally redundant or whether they serve distinct cellular roles. 12970363 6 69 theme glycosylation 1017:1029 arg1 sites 1031:1035 both extracellular N-linked glycosylation sites 989:1035 both extracellular N-linked glycosylation sites 989:1035 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 11 70 theme glycosylated 1655:1666 arg1 site 1668:1671 an additional second glycosylated site 1634:1671 an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 1634:1723 Reciprocally, engineering of an additional second glycosylated site in TRPC3 to mimic the glycosylation status in TRPC6 markedly reduced TRPC3 basal activity. 12970363 0 71 theme TRPC6 74:78 arg1 activity 88:95 basal TRPC3 and TRPC6 channel activity 58:95 basal TRPC3 and TRPC6 channel activity 58:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 6 72 theme extracellular 994:1006 arg1 sites 1031:1035 both extracellular N-linked glycosylation sites 989:1035 both extracellular N-linked glycosylation sites 989:1035 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 4 73 theme regulated 722:730 arg1 channel 732:738 a tightly regulated channel 712:738 a tightly regulated channel 712:738 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 4 73 theme regulated 722:730 arg1 TRPC6 703:707 TRPC6 703:707 TRPC6 703:707 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 5 74 theme molecular 763:771 arg1 correlates 773:782 potential molecular correlates 753:782 potential molecular correlates accounting for the functional difference 753:823 To identify potential molecular correlates accounting for the functional difference, we analyzed the glycosylation pattern of TRPC6 compared with TRPC3. 12970363 7 75 dep opposed 1237:1243 arg1 L 1333:1333 L 1333:1333 L 1333:1333 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 75 dep opposed 1237:1243 arg1 B. 1293:1294 B. 1293:1294 B. 1293:1294 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 75 dep opposed 1237:1243 arg1 Zhu 1297:1299 Zhu 1297:1299 Zhu 1297:1299 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 75 dep opposed 1237:1243 arg1 D. 1313:1314 D. 1313:1314 D. 1313:1314 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 75 dep opposed 1237:1243 arg1 1998 1337:1340 1998 1337:1340 1998 1337:1340 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 75 dep opposed 1237:1243 arg1 X. 1302:1303 X. 1302:1303 X. 1302:1303 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 75 dep opposed 1237:1243 arg1 Birnbaumer 1321:1330 Birnbaumer 1321:1330 Birnbaumer 1321:1330 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 2 76 theme common 312:317 arg1 characteristics 331:345 common biophysical characteristics 312:345 common biophysical characteristics 312:345 Members of the TRPC3/6/7 subfamily share common biophysical characteristics and are activated by diacylglycerol in a membrane-delimited manner. 12970363 7 77 theme first 1188:1192 arg1 e2 1230:1231 e2 1230:1231 e2 1230:1231 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 77 theme first 1188:1192 arg1 loop 1224:1227 the first (e1) and second extracellular loop 1184:1227 the first (e1) and second extracellular loop (e2) 1184:1232 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 2 78 theme membrane-delimited 388:405 arg1 manner 407:412 a membrane-delimited manner 386:412 a membrane-delimited manner 386:412 Members of the TRPC3/6/7 subfamily share common biophysical characteristics and are activated by diacylglycerol in a membrane-delimited manner. 12970363 0 79 theme protein 9:15 arg1 glycosylation 17:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 0 79 theme protein 9:15 arg1 determinant 42:52 a major determinant 34:52 a major determinant for basal TRPC3 and TRPC6 channel activity 34:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 5 80 theme TRPC6 867:871 arg1 pattern 856:862 the glycosylation pattern 838:862 the glycosylation pattern of TRPC6 838:871 To identify potential molecular correlates accounting for the functional difference, we analyzed the glycosylation pattern of TRPC6 compared with TRPC3. 12970363 7 81 gly glycosylated 1164:1175 arg1 model 1134:1138 a model 1132:1138 a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol 1132:1349 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 4 82 theme electrophysiological 576:595 arg1 strategies 622:631 electrophysiological and fluorescence imaging strategies 576:631 electrophysiological and fluorescence imaging strategies 576:631 By electrophysiological and fluorescence imaging strategies we show that TRPC3 displays considerable constitutive activity, while TRPC6 is a tightly regulated channel. 12970363 0 83 theme major 36:40 arg1 glycosylation 17:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation 0:29 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 0 83 theme major 36:40 arg1 determinant 42:52 a major determinant 34:52 a major determinant for basal TRPC3 and TRPC6 channel activity 34:95 N-linked protein glycosylation is a major determinant for basal TRPC3 and TRPC6 channel activity. 12970363 6 84 theme S/T 901:903 arg1 motifs 906:911 Two NX(S/T) motifs 894:911 Two NX(S/T) motifs in TRPC6 894:920 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 7 85 theme TRPC3 1269:1273 arg1 channel 1275:1281 the monoglycosylated TRPC3 channel 1248:1281 the monoglycosylated TRPC3 channel 1248:1281 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 7 86 gly monoglycosylated 1252:1267 arg1 channel 1275:1281 the monoglycosylated TRPC3 channel 1248:1281 the monoglycosylated TRPC3 channel 1248:1281 Immunoblotting analysis of HEK 293 cell lysates expressing TRPC6 wild type and mutants favors a model of TRPC6 that is dually glycosylated within the first (e1) and second extracellular loop (e2) as opposed to the monoglycosylated TRPC3 channel (Vannier, B., Zhu, X., Brown, D., and Birnbaumer, L. (1998) J. Biol. 12970363 6 87 theme NX 898:899 arg1 motifs 906:911 Two NX(S/T) motifs 894:911 Two NX(S/T) motifs in TRPC6 894:920 Two NX(S/T) motifs in TRPC6 were mutated (Asn to Gln) by in vitro mutagenesis to delete one or both extracellular N-linked glycosylation sites. 12970363 5 88 theme potential 753:761 arg1 correlates 773:782 potential molecular correlates 753:782 potential molecular correlates accounting for the functional difference 753:823 To identify potential molecular correlates accounting for the functional difference, we analyzed the glycosylation pattern of TRPC6 compared with TRPC3. 12970363 10 89 theme site 1411:1414 arg1 Elimination 1375:1385 Elimination 1375:1385 Elimination 1375:1385 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 10 90 theme e2 1394:1395 arg1 site 1411:1414 the e2 glycosylation site 1390:1414 the e2 glycosylation site 1390:1414 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 12970363 12 91 theme tight 1836:1840 arg1 regulation 1842:1851 the tight regulation 1832:1851 the tight regulation of TRPC6 through phospholipase C-activating receptors 1832:1905 We conclude that the glycosylation pattern plays a pivotal role for the tight regulation of TRPC6 through phospholipase C-activating receptors. 12970363 10 92 theme TRPC3 1598:1602 arg1 characteristics 1579:1593 functional characteristics 1568:1593 functional characteristics of TRPC3 1568:1602 Elimination of the e2 glycosylation site, missing in the monoglycosylated TRPC3, was sufficient to convert the tightly receptor-regulated TRPC6 into a constitutively active channel, displaying functional characteristics of TRPC3. 18070108 6 0 from conformations 842:854 arg1 BMP-6 859:863 BMP-6 859:863 BMP-6 859:863 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 6 1 theme crucial 755:761 arg1 role 763:766 a crucial role 753:766 a crucial role 753:766 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 2 2 theme related 318:324 arg1 proteins 335:342 more than 30 structurally related secreted proteins 292:342 more than 30 structurally related secreted proteins 292:342 This superfamily is formed by more than 30 structurally related secreted proteins. 18070108 10 3 theme BMP-6 1500:1504 arg1 epitope 1489:1495 the binding epitope 1477:1495 the binding epitope of BMP-6 1477:1504 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 7 4 theme promiscuous 1045:1055 arg1 binding 1057:1063 promiscuous binding 1045:1063 promiscuous binding of different type I receptors to BMPs 1045:1101 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 10 5 theme BMP-6 1637:1641 arg1 specificity 1622:1632 type I receptor specificity 1606:1632 type I receptor specificity of BMP-6 1606:1641 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 8 6 theme Further 1104:1110 arg1 studies 1112:1118 Further studies 1104:1118 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors 1104:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 3 7 theme superfamily 482:492 arg1 ligands 494:500 other TGF-beta superfamily ligands 467:500 other TGF-beta superfamily ligands 467:500 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 9 8 theme moiety 1369:1374 arg1 absence 1341:1347 the absence 1337:1347 the absence of the carbohydrate moiety 1337:1374 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 6 9 theme BMP-2 807:811 arg1 interactions 791:802 the type I receptor interactions 771:802 the type I receptor interactions of BMP-2 771:811 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 8 10 theme I 1194:1194 arg1 receptors 1196:1204 type I receptors 1189:1204 type I receptors 1189:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 10 11 theme recognition 1521:1531 arg1 motif 1564:1568 an N-glycosylation motif 1545:1568 i.e. an N-glycosylation motif 1540:1568 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 10 11 theme recognition 1521:1531 arg1 motif 1533:1537 an unusual recognition motif 1510:1537 an unusual recognition motif 1510:1537 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 7 12 theme important 1014:1022 arg1 feature 1024:1030 an important feature 1011:1030 an important feature required for promiscuous binding of different type I receptors to BMPs 1011:1101 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 7 12 theme important 1014:1022 arg1 Flexibility 956:966 Flexibility 956:966 Flexibility of this loop segment 956:987 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 6 13 theme prehelix 726:733 arg1 loop 735:738 the prehelix loop 722:738 the prehelix loop 722:738 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 9 14 theme activin 1377:1383 arg1 signaling 1410:1418 activin receptor type I-mediated signaling 1377:1418 activin receptor type I-mediated signaling of BMP-6 1377:1427 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 4 15 theme possible 579:586 arg1 asymmetry 588:596 possible asymmetry 579:596 possible asymmetry between the two monomeric subunits 579:631 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 9 16 theme type 1394:1397 arg1 signaling 1410:1418 activin receptor type I-mediated signaling 1377:1418 activin receptor type I-mediated signaling of BMP-6 1377:1427 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 8 17 theme BMP-6 1153:1157 arg1 interaction 1138:1148 the interaction 1134:1148 the interaction of BMP-6 with different ectodomains of type I receptors 1134:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 6 18 theme dynamic 886:892 arg1 flexibility 894:904 possible dynamic flexibility 877:904 possible dynamic flexibility of the prehelix loop in its unbound conformation 877:953 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 5 19 theme loops 670:674 arg1 conformation 646:657 the conformation 642:657 the conformation of several loops 642:674 Indeed, the conformation of several loops differs between both monomers. 18070108 8 20 theme different 1164:1172 arg1 ectodomains 1174:1184 different ectodomains 1164:1184 different ectodomains of type I receptors 1164:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 8 20 theme different 1164:1172 arg1 receptors 1196:1204 type I receptors 1189:1204 type I receptors 1189:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 7 21 theme segment 981:987 arg1 feature 1024:1030 an important feature 1011:1030 an important feature required for promiscuous binding of different type I receptors to BMPs 1011:1101 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 7 21 theme segment 981:987 arg1 Flexibility 956:966 Flexibility 956:966 Flexibility of this loop segment 956:987 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 4 22 theme monomeric 614:622 arg1 subunits 624:631 the two monomeric subunits 606:631 the two monomeric subunits 606:631 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 8 23 theme BMP-6 1248:1252 arg1 Asn73 1239:1243 Asn73 1239:1243 Asn73 of BMP-6 1239:1252 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 8 24 from N-glycosylation 1220:1234 arg1 epitope 1267:1273 the wrist epitope 1257:1273 the wrist epitope 1257:1273 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 3 25 dep BMP-7 508:512 arg1 e.g. 503:506 e.g. 503:506 e.g. 503:506 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 6 26 theme unbound 934:940 arg1 conformation 942:953 its unbound conformation 930:953 its unbound conformation 930:953 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 10 27 gly N-glycosylation 1548:1562 arg2 motif 1564:1568 an N-glycosylation motif 1545:1568 i.e. an N-glycosylation motif 1540:1568 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 10 27 gly N-glycosylation 1548:1562 arg2 motif 1533:1537 an unusual recognition motif 1510:1537 an unusual recognition motif 1510:1537 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 8 28 from Asn73 1239:1243 arg1 N-glycosylation 1220:1234 N-glycosylation 1220:1234 N-glycosylation at Asn73 of BMP-6 in the wrist epitope 1220:1273 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 3 29 dep structure 435:443 arg1 similar 448:454 similar 448:454 similar 448:454 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 6 30 theme prehelix 913:920 arg1 loop 922:925 the prehelix loop 909:925 the prehelix loop 909:925 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 0 31 theme bone 27:30 arg1 protein 46:52 bone morphogenetic protein 6 27:54 bone morphogenetic protein 6 27:54 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 0 32 theme Type 0:3 arg1 binding 16:22 Type I receptor binding 0:22 Type I receptor binding of bone morphogenetic protein 6 0:54 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 6 33 from flexibility 894:904 arg1 conformation 942:953 its unbound conformation 930:953 its unbound conformation 930:953 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 8 34 theme receptor 1317:1324 arg1 type 1326:1329 the activin receptor type I 1305:1331 the activin receptor type I 1305:1331 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 0 35 theme receptor 7:14 arg1 binding 16:22 Type I receptor binding 0:22 Type I receptor binding of bone morphogenetic protein 6 0:54 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 10 36 theme important 1588:1596 arg1 role 1598:1601 an important role 1585:1601 an important role 1585:1601 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 7 37 theme receptors 1085:1093 arg1 binding 1057:1063 promiscuous binding 1045:1063 promiscuous binding of different type I receptors to BMPs 1045:1101 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 6 38 theme receptor 782:789 arg1 interactions 791:802 the type I receptor interactions 771:802 the type I receptor interactions of BMP-2 771:811 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 3 39 theme human 370:374 arg1 BMP-6 376:380 human BMP-6 370:380 human BMP-6 370:380 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 1 40 dep together 139:146 arg1 with 148:151 with 148:151 with 148:151 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 0 41 theme morphogenetic 32:44 arg1 protein 46:52 bone morphogenetic protein 6 27:54 bone morphogenetic protein 6 27:54 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 6 42 theme type 775:778 arg1 receptor 782:789 type I receptor 775:789 the type I receptor interactions of BMP-2 771:811 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 10 43 theme I 1611:1611 arg1 specificity 1622:1632 type I receptor specificity 1606:1632 type I receptor specificity of BMP-6 1606:1641 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 4 44 theme full 548:551 arg1 BMP-6 561:565 the full dimeric BMP-6 544:565 the full dimeric BMP-6 544:565 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 10 45 theme binding 1481:1487 arg1 epitope 1489:1495 the binding epitope 1477:1495 the binding epitope of BMP-6 1477:1504 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 1 46 dep factor 173:178 arg1 -beta 185:189 -beta 185:189 -beta 185:189 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 0 47 theme protein 46:52 arg1 binding 16:22 Type I receptor binding 0:22 Type I receptor binding of bone morphogenetic protein 6 0:54 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 2 48 theme secreted 326:333 arg1 proteins 335:342 more than 30 structurally related secreted proteins 292:342 more than 30 structurally related secreted proteins 292:342 This superfamily is formed by more than 30 structurally related secreted proteins. 18070108 6 49 theme different 832:840 arg1 conformations 842:854 two rather different conformations 821:854 two rather different conformations in BMP-6 821:863 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 4 50 theme asymmetric 519:528 arg1 unit 530:533 The asymmetric unit 515:533 The asymmetric unit 515:533 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 1 51 theme TGF-beta 229:236 arg1 superfamily 238:248 the TGF-beta superfamily 225:248 the TGF-beta superfamily of ligands 225:259 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 3 52 theme TGF-beta 473:480 arg1 ligands 494:500 other TGF-beta superfamily ligands 467:500 other TGF-beta superfamily ligands 467:500 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 9 53 theme carbohydrate 1356:1367 arg1 moiety 1369:1374 the carbohydrate moiety 1352:1374 the carbohydrate moiety 1352:1374 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 7 54 theme type 1078:1081 arg1 receptors 1085:1093 different type I receptors 1068:1093 different type I receptors 1068:1093 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 1 55 theme transforming 153:164 arg1 TGF 181:183 TGF 181:183 TGF 181:183 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 1 55 theme transforming 153:164 arg1 factor 173:178 transforming growth factor 153:178 transforming growth factor (TGF) 153:184 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 8 56 theme type 1189:1192 arg1 receptors 1196:1204 type I receptors 1189:1204 type I receptors 1189:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 10 57 theme unusual 1513:1519 arg1 motif 1564:1568 an N-glycosylation motif 1545:1568 i.e. an N-glycosylation motif 1540:1568 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 10 57 theme unusual 1513:1519 arg1 motif 1533:1537 an unusual recognition motif 1510:1537 an unusual recognition motif 1510:1537 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 9 58 theme receptor 1385:1392 arg1 signaling 1410:1418 activin receptor type I-mediated signaling 1377:1418 activin receptor type I-mediated signaling of BMP-6 1377:1427 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 8 59 theme receptors 1196:1204 arg1 ectodomains 1174:1184 different ectodomains 1164:1184 different ectodomains of type I receptors 1164:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 8 59 theme receptors 1196:1204 arg1 receptors 1196:1204 type I receptors 1189:1204 type I receptors 1189:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 10 60 theme N-glycosylation 1548:1562 arg1 motif 1564:1568 an N-glycosylation motif 1545:1568 i.e. an N-glycosylation motif 1540:1568 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 10 60 theme N-glycosylation 1548:1562 arg1 motif 1533:1537 an unusual recognition motif 1510:1537 an unusual recognition motif 1510:1537 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 5 61 theme several 662:668 arg1 loops 670:674 several loops 662:674 several loops 662:674 Indeed, the conformation of several loops differs between both monomers. 18070108 4 62 contain contains 535:542 arg2 BMP-6 561:565 the full dimeric BMP-6 544:565 the full dimeric BMP-6 544:565 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 4 62 contain contains 535:542 arg1 unit 530:533 The asymmetric unit 515:533 The asymmetric unit 515:533 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 1 63 theme growth 166:171 arg1 TGF 181:183 TGF 181:183 TGF 181:183 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 1 63 theme growth 166:171 arg1 factor 173:178 transforming growth factor 153:178 transforming growth factor (TGF) 153:184 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 9 64 theme I-mediated 1399:1408 arg1 signaling 1410:1418 activin receptor type I-mediated signaling 1377:1418 activin receptor type I-mediated signaling of BMP-6 1377:1427 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 6 65 theme possible 877:884 arg1 flexibility 894:904 possible dynamic flexibility 877:904 possible dynamic flexibility of the prehelix loop in its unbound conformation 877:953 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 8 66 with interaction 1138:1148 arg1 ectodomains 1174:1184 different ectodomains 1164:1184 different ectodomains of type I receptors 1164:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 8 66 with interaction 1138:1148 arg1 receptors 1196:1204 type I receptors 1189:1204 type I receptors 1189:1204 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 7 67 theme loop 976:979 arg1 segment 981:987 this loop segment 971:987 this loop segment 971:987 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 1 68 theme morphogenetic 108:120 arg1 BMPs 132:135 BMPs 132:135 BMPs 132:135 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 1 68 theme morphogenetic 108:120 arg1 proteins 122:129 Bone morphogenetic proteins 103:129 Bone morphogenetic proteins (BMPs) 103:136 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 3 69 theme overall 427:433 arg1 structure 435:443 the overall structure 423:443 the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7 423:512 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 8 70 gly N-glycosylation 1220:1234 arg1 Asn73 1239:1243 Asn73 1239:1243 Asn73 of BMP-6 1239:1252 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 8 70 gly N-glycosylation 1220:1234 arg2 Asn73 1239:1243 Asn73 1239:1243 Asn73 of BMP-6 1239:1252 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 8 70 gly N-glycosylation 1220:1234 arg1 epitope 1267:1273 the wrist epitope 1257:1273 the wrist epitope 1257:1273 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 1 71 theme Bone 103:106 arg1 BMPs 132:135 BMPs 132:135 BMPs 132:135 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 1 71 theme Bone 103:106 arg1 proteins 122:129 Bone morphogenetic proteins 103:129 Bone morphogenetic proteins (BMPs) 103:136 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 8 72 theme wrist 1261:1265 arg1 epitope 1267:1273 the wrist epitope 1257:1273 the wrist epitope 1257:1273 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 6 73 theme loop 922:925 arg1 flexibility 894:904 possible dynamic flexibility 877:904 possible dynamic flexibility of the prehelix loop in its unbound conformation 877:953 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 0 74 theme ligand 95:100 arg1 N-glycosylation 72:86 N-glycosylation 72:86 N-glycosylation of the ligand 72:100 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 9 75 theme BMP-6 1423:1427 arg1 signaling 1410:1418 activin receptor type I-mediated signaling 1377:1418 activin receptor type I-mediated signaling of BMP-6 1377:1427 In the absence of the carbohydrate moiety, activin receptor type I-mediated signaling of BMP-6 is totally diminished. 18070108 0 76 theme I 5:5 arg1 binding 16:22 Type I receptor binding 0:22 Type I receptor binding of bone morphogenetic protein 6 0:54 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 0 77 gly N-glycosylation 72:86 arg1 ligand 95:100 the ligand 91:100 the ligand 91:100 Type I receptor binding of bone morphogenetic protein 6 is dependent on N-glycosylation of the ligand. 18070108 1 78 theme ligands 253:259 arg1 superfamily 238:248 the TGF-beta superfamily 225:248 the TGF-beta superfamily of ligands 225:259 Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)-beta and activins/inhibins, constitute the TGF-beta superfamily of ligands. 18070108 3 79 theme crystal 349:355 arg1 structure 357:365 The crystal structure 345:365 The crystal structure of human BMP-6 345:380 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 8 80 theme activin 1309:1315 arg1 type 1326:1329 the activin receptor type I 1305:1331 the activin receptor type I 1305:1331 Further studies investigating the interaction of BMP-6 with different ectodomains of type I receptors revealed that N-glycosylation at Asn73 of BMP-6 in the wrist epitope is crucial for recognition by the activin receptor type I. 18070108 7 81 theme I 1083:1083 arg1 receptors 1085:1093 different type I receptors 1068:1093 different type I receptors 1068:1093 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 3 82 theme A 420:420 arg1 resolution 402:411 a resolution 400:411 a resolution of 2.1 A 400:420 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 7 83 theme different 1068:1076 arg1 receptors 1085:1093 different type I receptors 1068:1093 different type I receptors 1068:1093 Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. 18070108 6 84 theme I 780:780 arg1 receptor 782:789 type I receptor 775:789 the type I receptor interactions of BMP-2 771:811 In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP-2, adopts two rather different conformations in BMP-6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. 18070108 3 85 theme BMP-6 376:380 arg1 structure 357:365 The crystal structure 345:365 The crystal structure of human BMP-6 345:380 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 18070108 10 86 theme receptor 1613:1620 arg1 specificity 1622:1632 type I receptor specificity 1606:1632 type I receptor specificity of BMP-6 1606:1641 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 4 87 theme dimeric 553:559 arg1 BMP-6 561:565 the full dimeric BMP-6 544:565 the full dimeric BMP-6 544:565 The asymmetric unit contains the full dimeric BMP-6, indicating possible asymmetry between the two monomeric subunits. 18070108 10 88 theme type 1606:1609 arg1 specificity 1622:1632 type I receptor specificity 1606:1632 type I receptor specificity of BMP-6 1606:1641 Thus, flexibility within the binding epitope of BMP-6 and an unusual recognition motif, i.e. an N-glycosylation motif, possibly play an important role in type I receptor specificity of BMP-6. 18070108 3 89 theme other 467:471 arg1 ligands 494:500 other TGF-beta superfamily ligands 467:500 other TGF-beta superfamily ligands 467:500 The crystal structure of human BMP-6 was determined to a resolution of 2.1 A; the overall structure is similar to that of other TGF-beta superfamily ligands, e.g. BMP-7. 21671571 0 0 theme active 100:105 arg1 center 107:112 the active center 96:112 the active center 96:112 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 5 1 from base 753:756 arg1 bond 741:744 a disulfide bond 729:744 a disulfide bond at the base of the active center that was not present in previous reports of hQC structure 729:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 1 from base 753:756 arg1 present 792:798 present 792:798 present 792:798 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 3 2 from structures 420:429 arg1 mQC 490:492 mQC 490:492 mQC 490:492 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 3 2 from structures 420:429 arg1 hQC 475:477 hQC 475:477 hQC 475:477 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 3 2 from structures 420:429 arg1 mice 484:487 mice 484:487 mice (mQC) 484:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 3 2 from structures 420:429 arg1 humans 467:472 humans 467:472 humans (hQC) 467:478 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 4 3 theme enzymes 534:540 arg1 similar 546:552 similar 546:552 similar 546:552 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 4 3 theme enzymes 534:540 arg1 structures 516:525 the overall structures 504:525 the overall structures of the enzymes 504:540 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 2 4 theme development 377:387 arg1 inhibitors 322:331 inhibitors 322:331 inhibitors of QCs 322:338 As the formation of pGlu-amyloid has been linked with Alzheimer's disease, inhibitors of QCs are currently the subject of intense development. 21671571 2 4 theme development 377:387 arg1 subject 358:364 the subject 354:364 the subject of intense development 354:387 As the formation of pGlu-amyloid has been linked with Alzheimer's disease, inhibitors of QCs are currently the subject of intense development. 21671571 7 5 theme previous 1046:1053 arg1 structure 1055:1063 previous structure 1046:1063 previous structure 1046:1063 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 5 6 located present 792:798 arg2 bond 741:744 a disulfide bond 729:744 a disulfide bond at the base of the active center that was not present in previous reports of hQC structure 729:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 6 located present 792:798 arg2 present 792:798 present 792:798 present 792:798 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 6 located present 792:798 arg1 reports 812:818 previous reports 803:818 previous reports of hQC structure 803:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 2 7 theme QCs 336:338 arg1 inhibitors 322:331 inhibitors 322:331 inhibitors of QCs 322:338 As the formation of pGlu-amyloid has been linked with Alzheimer's disease, inhibitors of QCs are currently the subject of intense development. 21671571 2 7 theme QCs 336:338 arg1 subject 358:364 the subject 354:364 the subject of intense development 354:387 As the formation of pGlu-amyloid has been linked with Alzheimer's disease, inhibitors of QCs are currently the subject of intense development. 21671571 7 8 from conformation 1081:1092 arg1 QCs 1132:1134 both glycosylated human and murine QCs 1097:1134 both glycosylated human and murine QCs 1097:1134 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 0 9 gly glycosylated 14:25 arg1 cyclases 48:55 glycosylated mammalian glutaminyl cyclases 14:55 glycosylated mammalian glutaminyl cyclases 14:55 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 9 10 theme hydrophobic 1489:1499 arg1 funnel 1501:1506 a hydrophobic funnel 1487:1506 a hydrophobic funnel 1487:1506 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 7 11 theme murine 1125:1130 arg1 QCs 1132:1134 both glycosylated human and murine QCs 1097:1134 both glycosylated human and murine QCs 1097:1134 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 6 12 theme Site-directed 838:850 arg1 mutagenesis 852:862 Site-directed mutagenesis 838:862 Site-directed mutagenesis 838:862 Site-directed mutagenesis suggests a structure-stabilizing role of the disulfide bond. 21671571 7 13 theme conserved 967:975 arg1 W 997:997 W 997:997 W(207) 997:1002 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 7 13 theme conserved 967:975 arg1 residue 988:994 the conserved tryptophan residue 963:994 the conserved tryptophan residue 963:994 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 3 14 from mice 484:487 arg1 structures 420:429 three crystal structures 406:429 three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 406:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 3 14 from mice 484:487 arg1 QC 459:460 N-glycosylated mammalian QC 434:460 N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 434:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 8 15 theme inhibitors 1263:1272 arg1 constants 1250:1258 the binding constants 1238:1258 the binding constants of inhibitors such as the highly potent PQ50 (PBD150) 1238:1312 Although mutagenesis of W(207) into leucine or glutamine altered substrate conversion significantly, the binding constants of inhibitors such as the highly potent PQ50 (PBD150) were minimally affected. 21671571 9 16 theme crystal 1343:1349 arg1 structure 1351:1359 The crystal structure 1339:1359 The crystal structure of PQ50 bound to the active center of murine QC 1339:1407 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 5 17 theme conserved 696:704 arg1 residues 715:722 two conserved cysteine residues 692:722 two conserved cysteine residues 692:722 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 7 18 theme multiple 1021:1028 arg1 orientations 1030:1041 multiple orientations 1021:1041 multiple orientations 1021:1041 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 8 19 theme substrate 1202:1210 arg1 conversion 1212:1221 substrate conversion 1202:1221 substrate conversion 1202:1221 Although mutagenesis of W(207) into leucine or glutamine altered substrate conversion significantly, the binding constants of inhibitors such as the highly potent PQ50 (PBD150) were minimally affected. 21671571 2 20 theme pGlu-amyloid 267:278 arg1 formation 254:262 the formation 250:262 the formation of pGlu-amyloid 250:278 As the formation of pGlu-amyloid has been linked with Alzheimer's disease, inhibitors of QCs are currently the subject of intense development. 21671571 9 21 theme PQ50 1364:1367 arg1 structure 1351:1359 The crystal structure 1339:1359 The crystal structure of PQ50 bound to the active center of murine QC 1339:1407 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 1 22 theme glutaminyl 171:180 arg1 precursors 194:203 glutaminyl or glutamyl precursors 171:203 glutaminyl or glutamyl precursors 171:203 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 5 23 theme cysteine 706:713 arg1 residues 715:722 two conserved cysteine residues 692:722 two conserved cysteine residues 692:722 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 8 24 theme binding 1242:1248 arg1 constants 1250:1258 the binding constants 1238:1258 the binding constants of inhibitors such as the highly potent PQ50 (PBD150) 1238:1312 Although mutagenesis of W(207) into leucine or glutamine altered substrate conversion significantly, the binding constants of inhibitors such as the highly potent PQ50 (PBD150) were minimally affected. 21671571 7 25 theme single 1074:1079 arg1 conformation 1081:1092 a single conformation 1072:1092 a single conformation in both glycosylated human and murine QCs 1072:1134 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 9 26 theme principal 1417:1425 arg1 determinants 1435:1446 principal binding determinants 1417:1446 principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel 1417:1506 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 1 27 dep pyroglutamate 139:151 arg1 pE 162:163 pE 162:163 pE 162:163 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 1 27 dep pyroglutamate 139:151 arg1 pGlu 154:157 pGlu 154:157 pGlu 154:157 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 10 28 contain containing 1569:1578 arg1 QCs 1565:1567 two mammalian QCs 1551:1567 two mammalian QCs containing typical, conserved post-translational modifications 1551:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 10 28 contain containing 1569:1578 arg2 modifications 1618:1630 typical, conserved post-translational modifications 1580:1630 typical, conserved post-translational modifications 1580:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 2 29 theme intense 369:375 arg1 development 377:387 intense development 369:387 intense development 369:387 As the formation of pGlu-amyloid has been linked with Alzheimer's disease, inhibitors of QCs are currently the subject of intense development. 21671571 7 30 theme glycosylated 1102:1113 arg1 QCs 1132:1134 both glycosylated human and murine QCs 1097:1134 both glycosylated human and murine QCs 1097:1134 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 5 31 theme disulfide 731:739 arg1 bond 741:744 a disulfide bond 729:744 a disulfide bond at the base of the active center that was not present in previous reports of hQC structure 729:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 31 theme disulfide 731:739 arg1 present 792:798 present 792:798 present 792:798 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 3 32 from humans 467:472 arg1 structures 420:429 three crystal structures 406:429 three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 406:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 3 32 from humans 467:472 arg1 QC 459:460 N-glycosylated mammalian QC 434:460 N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 434:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 8 33 theme potent 1293:1298 arg1 PBD150 1306:1311 PBD150 1306:1311 PBD150 1306:1311 Although mutagenesis of W(207) into leucine or glutamine altered substrate conversion significantly, the binding constants of inhibitors such as the highly potent PQ50 (PBD150) were minimally affected. 21671571 8 33 theme potent 1293:1298 arg1 PQ50 1300:1303 the highly potent PQ50 1282:1303 the highly potent PQ50 (PBD150) 1282:1312 Although mutagenesis of W(207) into leucine or glutamine altered substrate conversion significantly, the binding constants of inhibitors such as the highly potent PQ50 (PBD150) were minimally affected. 21671571 1 34 theme glutamyl 185:192 arg1 precursors 194:203 glutaminyl or glutamyl precursors 171:203 glutaminyl or glutamyl precursors 171:203 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 7 35 theme human 1115:1119 arg1 QCs 1132:1134 both glycosylated human and murine QCs 1097:1134 both glycosylated human and murine QCs 1097:1134 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 4 36 theme overall 508:514 arg1 similar 546:552 similar 546:552 similar 546:552 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 4 36 theme overall 508:514 arg1 structures 516:525 the overall structures 504:525 the overall structures of the enzymes 504:540 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 10 37 theme post-translational 1599:1616 arg1 modifications 1618:1630 typical, conserved post-translational modifications 1580:1630 typical, conserved post-translational modifications 1580:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 6 38 theme bond 919:922 arg1 role 897:900 a structure-stabilizing role 873:900 a structure-stabilizing role of the disulfide bond 873:922 Site-directed mutagenesis suggests a structure-stabilizing role of the disulfide bond. 21671571 4 39 from loops 596:600 arg1 neighborhood 609:620 the neighborhood 605:620 the neighborhood of the active center 605:641 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 0 40 theme mammalian 27:35 arg1 cyclases 48:55 glycosylated mammalian glutaminyl cyclases 14:55 glycosylated mammalian glutaminyl cyclases 14:55 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 10 41 dep typical 1580:1586 arg1 conserved 1589:1597 conserved 1589:1597 conserved 1589:1597 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 3 42 theme mammalian 449:457 arg1 QC 459:460 N-glycosylated mammalian QC 434:460 N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 434:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 0 43 theme glycosylated 14:25 arg1 cyclases 48:55 glycosylated mammalian glutaminyl cyclases 14:55 glycosylated mammalian glutaminyl cyclases 14:55 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 10 44 theme mammalian 1555:1563 arg1 QCs 1565:1567 two mammalian QCs 1551:1567 two mammalian QCs containing typical, conserved post-translational modifications 1551:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 9 45 theme catalytic 1464:1472 arg1 ion 1479:1481 the catalytic zinc ion 1460:1481 the catalytic zinc ion 1460:1481 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 4 46 theme center 636:641 arg1 neighborhood 609:620 the neighborhood 605:620 the neighborhood of the active center 605:641 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 3 47 theme QC 459:460 arg1 structures 420:429 three crystal structures 406:429 three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 406:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 0 48 theme cyclases 48:55 arg1 Structures 0:9 Structures 0:9 Structures of glycosylated mammalian glutaminyl cyclases 0:55 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 4 49 theme conformational 651:664 arg1 variability 666:676 conformational variability 651:676 conformational variability 651:676 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 9 50 theme murine 1399:1404 arg1 QC 1406:1407 murine QC 1399:1407 murine QC 1399:1407 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 10 51 theme typical 1580:1586 arg1 modifications 1618:1630 typical, conserved post-translational modifications 1580:1630 typical, conserved post-translational modifications 1580:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 4 52 theme surface 588:594 arg1 loops 596:600 two surface loops 584:600 two surface loops in the neighborhood of the active center 584:641 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 9 53 theme active 1382:1387 arg1 center 1389:1394 the active center 1378:1394 the active center of murine QC 1378:1407 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 0 54 theme glutaminyl 37:46 arg1 cyclases 48:55 glycosylated mammalian glutaminyl cyclases 14:55 glycosylated mammalian glutaminyl cyclases 14:55 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 6 55 theme disulfide 909:917 arg1 bond 919:922 the disulfide bond 905:922 the disulfide bond 905:922 Site-directed mutagenesis suggests a structure-stabilizing role of the disulfide bond. 21671571 10 56 theme first 1531:1535 arg1 comparison 1537:1546 a first comparison 1529:1546 a first comparison of two mammalian QCs containing typical, conserved post-translational modifications 1529:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 5 57 theme active 765:770 arg1 center 772:777 the active center 761:777 the active center 761:777 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 1 58 theme glutaminyl 221:230 arg1 QC 242:243 QC 242:243 QC 242:243 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 1 58 theme glutaminyl 221:230 arg1 cyclases 232:239 glutaminyl cyclases 221:239 glutaminyl cyclases (QC) 221:244 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 0 59 theme conformational 64:77 arg1 variability 79:89 conformational variability 64:89 conformational variability 64:89 Structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center. 21671571 5 60 theme hQC 823:825 arg1 structure 827:835 hQC structure 823:835 hQC structure 823:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 3 61 theme crystal 412:418 arg1 structures 420:429 three crystal structures 406:429 three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 406:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 3 62 gly N-glycosylated 434:447 arg1 QC 459:460 N-glycosylated mammalian QC 434:460 N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 434:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 5 63 from present 792:798 arg1 reports 812:818 previous reports 803:818 previous reports of hQC structure 803:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 1 64 theme pyroglutamate 139:151 arg1 Formation 115:123 Formation 115:123 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors 115:203 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 1 65 from Formation 115:123 arg1 precursors 194:203 glutaminyl or glutamyl precursors 171:203 glutaminyl or glutamyl precursors 171:203 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 7 66 theme active 948:953 arg1 center 955:960 the active center 944:960 the active center 944:960 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 7 67 theme tryptophan 977:986 arg1 W 997:997 W 997:997 W(207) 997:1002 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 7 67 theme tryptophan 977:986 arg1 residue 988:994 the conserved tryptophan residue 963:994 the conserved tryptophan residue 963:994 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 9 68 theme QC 1406:1407 arg1 center 1389:1394 the active center 1378:1394 the active center of murine QC 1378:1407 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 9 69 theme zinc 1474:1477 arg1 ion 1479:1481 the catalytic zinc ion 1460:1481 the catalytic zinc ion 1460:1481 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 5 70 attach present 792:798 arg2 bond 741:744 a disulfide bond 729:744 a disulfide bond at the base of the active center that was not present in previous reports of hQC structure 729:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 70 attach present 792:798 arg2 present 792:798 present 792:798 present 792:798 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 70 attach present 792:798 arg1 reports 812:818 previous reports 803:818 previous reports of hQC structure 803:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 7 71 gly glycosylated 1102:1113 arg1 QCs 1132:1134 both glycosylated human and murine QCs 1097:1134 both glycosylated human and murine QCs 1097:1134 At the entrance to the active center, the conserved tryptophan residue, W(207), which displayed multiple orientations in previous structure, shows a single conformation in both glycosylated human and murine QCs. 21671571 5 72 from reports 812:818 arg1 bond 741:744 a disulfide bond 729:744 a disulfide bond at the base of the active center that was not present in previous reports of hQC structure 729:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 72 from reports 812:818 arg1 present 792:798 present 792:798 present 792:798 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 9 73 theme binding 1427:1433 arg1 determinants 1435:1446 principal binding determinants 1417:1446 principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel 1417:1506 The crystal structure of PQ50 bound to the active center of murine QC reveals principal binding determinants provided by the catalytic zinc ion and a hydrophobic funnel. 21671571 3 74 theme N-glycosylated 434:447 arg1 QC 459:460 N-glycosylated mammalian QC 434:460 N-glycosylated mammalian QC from humans (hQC) and mice (mQC) 434:493 Here, we report three crystal structures of N-glycosylated mammalian QC from humans (hQC) and mice (mQC). 21671571 1 75 from precursors 194:203 arg1 Formation 115:123 Formation 115:123 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors 115:203 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 1 75 from precursors 194:203 arg1 pyroglutamate 139:151 pyroglutamate 139:151 pyroglutamate 139:151 Formation of N-terminal pyroglutamate (pGlu or pE) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (QC). 21671571 4 76 theme active 629:634 arg1 center 636:641 the active center 625:641 the active center 625:641 Whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighborhood of the active center exhibit conformational variability. 21671571 5 77 theme previous 803:810 arg1 reports 812:818 previous reports 803:818 previous reports of hQC structure 803:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 10 78 theme QCs 1565:1567 arg1 comparison 1537:1546 a first comparison 1529:1546 a first comparison of two mammalian QCs containing typical, conserved post-translational modifications 1529:1630 This study presents a first comparison of two mammalian QCs containing typical, conserved post-translational modifications. 21671571 8 79 theme W 1161:1161 arg1 mutagenesis 1146:1156 mutagenesis 1146:1156 mutagenesis of W(207) into leucine or glutamine 1146:1192 Although mutagenesis of W(207) into leucine or glutamine altered substrate conversion significantly, the binding constants of inhibitors such as the highly potent PQ50 (PBD150) were minimally affected. 21671571 6 80 theme structure-stabilizing 875:895 arg1 role 897:900 a structure-stabilizing role 873:900 a structure-stabilizing role of the disulfide bond 873:922 Site-directed mutagenesis suggests a structure-stabilizing role of the disulfide bond. 21671571 5 81 theme center 772:777 arg1 base 753:756 the base 749:756 the base of the active center 749:777 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21671571 5 82 theme structure 827:835 arg1 reports 812:818 previous reports 803:818 previous reports of hQC structure 803:835 Furthermore, two conserved cysteine residues form a disulfide bond at the base of the active center that was not present in previous reports of hQC structure. 21199866 0 0 theme ATP-binding 79:89 arg1 transporters 106:117 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 0 theme ATP-binding 79:89 arg1 ABCB6 119:123 ABCB6 119:123 ABCB6 119:123 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 0 theme ATP-binding 79:89 arg1 /ABCC8 159:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 12 1 theme mutant 1541:1546 arg1 allele 1548:1553 The mutant allele 1537:1553 The mutant allele 1537:1553 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 4 2 gly glycosylated 534:545 arg1 ABCB6 525:529 endogenous ABCB6 514:529 endogenous ABCB6 514:529 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 4 2 gly glycosylated 534:545 arg2 ABCB6 525:529 endogenous ABCB6 514:529 endogenous ABCB6 514:529 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 4 2 gly glycosylated 534:545 arg1 types 564:568 multiple cell types 550:568 multiple cell types 550:568 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 12 3 theme amino-terminal 1595:1608 arg1 cysteine 1610:1617 a conserved amino-terminal cysteine 1583:1617 a conserved amino-terminal cysteine 1583:1617 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 7 4 theme additional 1014:1023 arg1 Cys-26 1063:1068 Cys-26 1063:1068 Cys-26 1063:1068 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 7 4 theme additional 1014:1023 arg1 cysteine 1053:1060 an additional highly conserved ER luminal cysteine 1011:1060 an additional highly conserved ER luminal cysteine (Cys-26) 1011:1069 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 0 5 dep transporters 106:117 arg1 transporters 106:117 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 5 dep transporters 106:117 arg1 ABCB6 119:123 ABCB6 119:123 ABCB6 119:123 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 5 dep transporters 106:117 arg1 /ABCC8 159:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 11 6 theme disease-causing 1417:1431 arg1 mutation 1433:1440 a disease-causing mutation 1415:1440 a disease-causing mutation 1415:1440 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 1 7 theme mitochondrial 221:233 arg1 ABCB6 210:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 1 7 theme mitochondrial 221:233 arg1 transporter 245:255 a mitochondrial porphyrin transporter 219:255 a mitochondrial porphyrin transporter that activates porphyrin biosynthesis 219:293 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 10 8 theme conserved 1356:1364 arg1 cysteines 1366:1374 conserved cysteines 1356:1374 conserved cysteines 1356:1374 We discovered that other ABC transporters with an amino terminus in the ER had similarly configured conserved cysteines. 21199866 11 9 theme sulfonylurea 1449:1460 arg1 receptor 1462:1469 sulfonylurea receptor 1 1449:1471 the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia 1445:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 11 9 theme sulfonylurea 1449:1460 arg1 SUR1 1474:1477 SUR1 1474:1477 SUR1 1474:1477 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 4 10 from site 660:663 arg1 terminus 702:709 its amino terminus 692:709 its amino terminus 692:709 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 1 11 theme porphyrin 235:243 arg1 ABCB6 210:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 1 11 theme porphyrin 235:243 arg1 transporter 245:255 a mitochondrial porphyrin transporter 219:255 a mitochondrial porphyrin transporter that activates porphyrin biosynthesis 219:293 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 0 12 theme cassette 91:98 arg1 transporters 106:117 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 12 theme cassette 91:98 arg1 ABCB6 119:123 ABCB6 119:123 ABCB6 119:123 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 12 theme cassette 91:98 arg1 /ABCC8 159:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 10 13 theme amino 1306:1310 arg1 terminus 1312:1319 an amino terminus 1303:1319 an amino terminus in the ER 1303:1329 We discovered that other ABC transporters with an amino terminus in the ER had similarly configured conserved cysteines. 21199866 10 14 from terminus 1312:1319 arg1 ER 1328:1329 the ER 1324:1329 the ER 1324:1329 We discovered that other ABC transporters with an amino terminus in the ER had similarly configured conserved cysteines. 21199866 13 15 theme ABC 1704:1706 arg1 transporters 1708:1719 ABC transporters 1704:1719 ABC transporters 1704:1719 These results suggest that for ABC transporters the propensity to form a disulfide bond in the ER defines a unique checkpoint that determines whether a protein is ER-retained. 21199866 7 16 theme ER 1042:1043 arg1 Cys-26 1063:1068 Cys-26 1063:1068 Cys-26 1063:1068 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 7 16 theme ER 1042:1043 arg1 cysteine 1053:1060 an additional highly conserved ER luminal cysteine 1011:1060 an additional highly conserved ER luminal cysteine (Cys-26) 1011:1069 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 2 17 theme plasma 420:425 arg1 membrane 427:434 the plasma membrane 416:434 the plasma membrane 416:434 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 4 18 theme atypical 651:658 arg1 site 660:663 only one atypical site 642:663 only one atypical site for glycosylation (NXC) in its amino terminus 642:709 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 0 19 theme ABC 101:103 arg1 transporters 106:117 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 19 theme ABC 101:103 arg1 ABCB6 119:123 ABCB6 119:123 ABCB6 119:123 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 19 theme ABC 101:103 arg1 /ABCC8 159:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 9 20 theme disulfide 1240:1248 arg1 bond 1250:1253 a disulfide bond 1238:1253 a disulfide bond 1238:1253 Further analysis revealed that these two cysteines form a disulfide bond. 21199866 7 21 theme luminal 1045:1051 arg1 Cys-26 1063:1068 Cys-26 1063:1068 Cys-26 1063:1068 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 7 21 theme luminal 1045:1051 arg1 cysteine 1053:1060 an additional highly conserved ER luminal cysteine 1011:1060 an additional highly conserved ER luminal cysteine (Cys-26) 1011:1069 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 12 22 from mutation 1571:1578 arg1 cysteine 1610:1617 a conserved amino-terminal cysteine 1583:1617 a conserved amino-terminal cysteine 1583:1617 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 10 23 theme other 1275:1279 arg1 transporters 1285:1296 other ABC transporters 1275:1296 other ABC transporters with an amino terminus in the ER 1275:1329 We discovered that other ABC transporters with an amino terminus in the ER had similarly configured conserved cysteines. 21199866 5 24 theme consensus 821:829 arg1 NXS 837:839 NXS 837:839 NXS 837:839 ABCB6 remained glycosylated when the highly conserved cysteine (Cys-8) was substituted with serine to make a consensus site, NXS. 21199866 5 24 theme consensus 821:829 arg1 site 831:834 a consensus site 819:834 a consensus site 819:834 ABCB6 remained glycosylated when the highly conserved cysteine (Cys-8) was substituted with serine to make a consensus site, NXS. 21199866 0 25 theme intramolecular 10:23 arg1 bond 35:38 Conserved intramolecular disulfide bond 0:38 Conserved intramolecular disulfide bond 0:38 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 2 26 theme cellular 386:393 arg1 compartments 395:406 other cellular compartments 380:406 other cellular compartments such as the plasma membrane 380:434 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 2 26 theme cellular 386:393 arg1 membrane 427:434 the plasma membrane 416:434 the plasma membrane 416:434 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 0 27 theme Conserved 0:8 arg1 bond 35:38 Conserved intramolecular disulfide bond 0:38 Conserved intramolecular disulfide bond 0:38 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 4 28 theme multiple 550:557 arg1 types 564:568 multiple cell types 550:568 multiple cell types 550:568 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 7 29 theme conserved 1032:1040 arg1 Cys-26 1063:1068 Cys-26 1063:1068 Cys-26 1063:1068 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 7 29 theme conserved 1032:1040 arg1 cysteine 1053:1060 an additional highly conserved ER luminal cysteine 1011:1060 an additional highly conserved ER luminal cysteine (Cys-26) 1011:1069 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 4 30 theme amino 696:700 arg1 terminus 702:709 its amino terminus 692:709 its amino terminus 692:709 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 7 31 contain has 1007:1009 arg1 terminus 989:996 The amino terminus 979:996 The amino terminus of ABCB6 979:1005 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 7 31 contain has 1007:1009 arg2 Cys-26 1063:1068 Cys-26 1063:1068 Cys-26 1063:1068 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 7 31 contain has 1007:1009 arg2 cysteine 1053:1060 an additional highly conserved ER luminal cysteine 1011:1060 an additional highly conserved ER luminal cysteine (Cys-26) 1011:1069 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 2 32 theme other 380:384 arg1 compartments 395:406 other cellular compartments 380:406 other cellular compartments such as the plasma membrane 380:434 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 2 32 theme other 380:384 arg1 membrane 427:434 the plasma membrane 416:434 the plasma membrane 416:434 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 11 33 theme receptor 1462:1469 arg1 /ABCC8 1479:1484 the sulfonylurea receptor 1 (SUR1)/ABCC8 1445:1484 the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia 1445:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 13 34 theme disulfide 1746:1754 arg1 bond 1756:1759 a disulfide bond 1744:1759 a disulfide bond 1744:1759 These results suggest that for ABC transporters the propensity to form a disulfide bond in the ER defines a unique checkpoint that determines whether a protein is ER-retained. 21199866 1 35 theme ATP-binding 171:181 arg1 cassette 183:190 ATP-binding cassette 171:190 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 1 35 theme ATP-binding 171:181 arg1 ABC 193:195 ABC 193:195 ABC 193:195 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 0 36 theme disulfide 25:33 arg1 bond 35:38 Conserved intramolecular disulfide bond 0:38 Conserved intramolecular disulfide bond 0:38 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 1 37 theme porphyrin 272:280 arg1 biosynthesis 282:293 porphyrin biosynthesis 272:293 porphyrin biosynthesis 272:293 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 0 38 theme transporters 106:117 arg1 fate 71:74 fate 71:74 fate 71:74 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 38 theme transporters 106:117 arg1 trafficking 55:65 trafficking 55:65 trafficking 55:65 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 1 39 theme cassette 183:190 arg1 ABCB6 210:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 1 39 theme cassette 183:190 arg1 transporter 245:255 a mitochondrial porphyrin transporter 219:255 a mitochondrial porphyrin transporter that activates porphyrin biosynthesis 219:293 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 4 40 theme cell 559:562 arg1 types 564:568 multiple cell types 550:568 multiple cell types 550:568 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 0 41 theme sulfonylurea 129:140 arg1 receptor 142:149 sulfonylurea receptor 1 129:151 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 41 theme sulfonylurea 129:140 arg1 SUR1 154:157 SUR1 154:157 SUR1 154:157 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 8 42 theme ER 1168:1169 arg1 retention 1171:1179 ER retention 1168:1179 ER retention 1168:1179 When Cys-26 was mutated alone or in combination with Cys-8, it also resulted in instability and ER retention. 21199866 5 43 theme conserved 756:764 arg1 cysteine 766:773 the highly conserved cysteine 745:773 the highly conserved cysteine (Cys-8) 745:781 ABCB6 remained glycosylated when the highly conserved cysteine (Cys-8) was substituted with serine to make a consensus site, NXS. 21199866 5 43 theme conserved 756:764 arg1 Cys-8 776:780 Cys-8 776:780 Cys-8 776:780 ABCB6 remained glycosylated when the highly conserved cysteine (Cys-8) was substituted with serine to make a consensus site, NXS. 21199866 6 44 theme ABCB6 898:902 arg1 degradation 904:914 ABCB6 degradation 898:914 ABCB6 degradation 898:914 However, this substitution blocked ER exit and produced ABCB6 degradation, which was mostly reversed by the proteasomal inhibitor MG132. 21199866 11 45 with patient 1493:1499 arg1 hypoglycemia 1523:1534 hyperinsulinemic hypoglycemia 1506:1534 hyperinsulinemic hypoglycemia 1506:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 4 46 theme endogenous 514:523 arg1 ABCB6 525:529 endogenous ABCB6 514:529 endogenous ABCB6 514:529 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 11 47 from discovery 1402:1410 arg1 /ABCC8 1479:1484 the sulfonylurea receptor 1 (SUR1)/ABCC8 1445:1484 the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia 1445:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 11 47 from discovery 1402:1410 arg1 patient 1493:1499 a patient 1491:1499 a patient with hyperinsulinemic hypoglycemia 1491:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 11 48 theme hyperinsulinemic 1506:1521 arg1 hypoglycemia 1523:1534 hyperinsulinemic hypoglycemia 1506:1534 hyperinsulinemic hypoglycemia 1506:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 10 49 with transporters 1285:1296 arg1 terminus 1312:1319 an amino terminus 1303:1319 an amino terminus in the ER 1303:1329 We discovered that other ABC transporters with an amino terminus in the ER had similarly configured conserved cysteines. 21199866 0 50 theme receptor 142:149 arg1 transporters 106:117 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8 79:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 0 50 theme receptor 142:149 arg1 /ABCC8 159:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 sulfonylurea receptor 1 (SUR1)/ABCC8 129:164 Conserved intramolecular disulfide bond is critical to trafficking and fate of ATP-binding cassette (ABC) transporters ABCB6 and sulfonylurea receptor 1 (SUR1)/ABCC8. 21199866 2 51 theme targeting 334:342 arg1 sequence 344:351 a canonical mitochondrial targeting sequence 308:351 a canonical mitochondrial targeting sequence 308:351 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 12 52 contain contains 1560:1567 arg1 allele 1548:1553 The mutant allele 1537:1553 The mutant allele 1537:1553 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 12 52 contain contains 1560:1567 arg2 mutation 1571:1578 a mutation 1569:1578 a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface 1569:1670 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 12 53 theme cell 1659:1662 arg1 surface 1664:1670 the cell surface 1655:1670 the cell surface 1655:1670 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 9 54 theme Further 1182:1188 arg1 analysis 1190:1197 Further analysis 1182:1197 Further analysis 1182:1197 Further analysis revealed that these two cysteines form a disulfide bond. 21199866 8 55 with combination 1108:1118 arg1 Cys-8 1125:1129 Cys-8 1125:1129 Cys-8 1125:1129 When Cys-26 was mutated alone or in combination with Cys-8, it also resulted in instability and ER retention. 21199866 2 56 theme mitochondrial 320:332 arg1 sequence 344:351 a canonical mitochondrial targeting sequence 308:351 a canonical mitochondrial targeting sequence 308:351 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 12 57 theme conserved 1585:1593 arg1 cysteine 1610:1617 a conserved amino-terminal cysteine 1583:1617 a conserved amino-terminal cysteine 1583:1617 The mutant allele only contains a mutation in a conserved amino-terminal cysteine, producing SUR1 that fails to reach the cell surface. 21199866 4 58 theme endoplasmic 606:616 arg1 ER 629:630 ER 629:630 ER 629:630 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 4 58 theme endoplasmic 606:616 arg1 reticulum 618:626 the endoplasmic reticulum 602:626 the endoplasmic reticulum (ER) 602:631 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 7 59 theme amino 983:987 arg1 terminus 989:996 The amino terminus 979:996 The amino terminus of ABCB6 979:1005 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 4 60 contain has 638:640 arg2 site 660:663 only one atypical site 642:663 only one atypical site for glycosylation (NXC) in its amino terminus 642:709 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 4 60 contain has 638:640 arg1 ABCB6 525:529 endogenous ABCB6 514:529 endogenous ABCB6 514:529 In this study, we show that endogenous ABCB6 is glycosylated in multiple cell types, indicating trafficking through the endoplasmic reticulum (ER), and has only one atypical site for glycosylation (NXC) in its amino terminus. 21199866 2 61 theme canonical 310:318 arg1 sequence 344:351 a canonical mitochondrial targeting sequence 308:351 a canonical mitochondrial targeting sequence 308:351 ABCB6 lacks a canonical mitochondrial targeting sequence but reportedly traffics to other cellular compartments such as the plasma membrane. 21199866 1 62 theme transporter 198:208 arg1 ABCB6 210:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 167:214 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 1 62 theme transporter 198:208 arg1 transporter 245:255 a mitochondrial porphyrin transporter 219:255 a mitochondrial porphyrin transporter that activates porphyrin biosynthesis 219:293 The ATP-binding cassette (ABC) transporter ABCB6 is a mitochondrial porphyrin transporter that activates porphyrin biosynthesis. 21199866 11 63 theme mutation 1433:1440 arg1 discovery 1402:1410 the discovery 1398:1410 the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia 1398:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 13 64 theme unique 1781:1786 arg1 checkpoint 1788:1797 a unique checkpoint 1779:1797 a unique checkpoint that determines whether a protein is ER-retained 1779:1846 These results suggest that for ABC transporters the propensity to form a disulfide bond in the ER defines a unique checkpoint that determines whether a protein is ER-retained. 21199866 6 65 theme inhibitor 962:970 arg1 MG132 972:976 the proteasomal inhibitor MG132 946:976 the proteasomal inhibitor MG132 946:976 However, this substitution blocked ER exit and produced ABCB6 degradation, which was mostly reversed by the proteasomal inhibitor MG132. 21199866 6 66 theme ER 877:878 arg1 exit 880:883 ER exit 877:883 ER exit 877:883 However, this substitution blocked ER exit and produced ABCB6 degradation, which was mostly reversed by the proteasomal inhibitor MG132. 21199866 7 67 theme ABCB6 1001:1005 arg1 terminus 989:996 The amino terminus 979:996 The amino terminus of ABCB6 979:1005 The amino terminus of ABCB6 has an additional highly conserved ER luminal cysteine (Cys-26). 21199866 10 68 theme ABC 1281:1283 arg1 transporters 1285:1296 other ABC transporters 1275:1296 other ABC transporters with an amino terminus in the ER 1275:1329 We discovered that other ABC transporters with an amino terminus in the ER had similarly configured conserved cysteines. 21199866 11 69 from patient 1493:1499 arg1 /ABCC8 1479:1484 the sulfonylurea receptor 1 (SUR1)/ABCC8 1445:1484 the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia 1445:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 11 69 from patient 1493:1499 arg1 discovery 1402:1410 the discovery 1398:1410 the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia 1398:1534 This analysis led to the discovery of a disease-causing mutation in the sulfonylurea receptor 1 (SUR1)/ABCC8 from a patient with hyperinsulinemic hypoglycemia. 21199866 6 70 theme proteasomal 950:960 arg1 MG132 972:976 the proteasomal inhibitor MG132 946:976 the proteasomal inhibitor MG132 946:976 However, this substitution blocked ER exit and produced ABCB6 degradation, which was mostly reversed by the proteasomal inhibitor MG132. 21056543 0 0 theme blood 64:68 arg1 inhibitor 80:88 human blood protein C inhibitor 58:88 human blood protein C inhibitor 58:88 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 4 1 theme blood 498:502 arg1 plasma 504:509 human blood plasma 492:509 human blood plasma 492:509 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 1 2 gly glycoprotein 129:140 arg1 inhibitor 101:109 Protein C inhibitor 91:109 Protein C inhibitor (PCI) 91:115 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 1 2 gly glycoprotein 129:140 arg1 glycoprotein 129:140 a 57-kDa glycoprotein 120:140 a 57-kDa glycoprotein that exists in many tissues and secretions in human 120:192 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 0 3 theme human 58:62 arg1 inhibitor 80:88 human blood protein C inhibitor 58:88 human blood protein C inhibitor 58:88 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 5 4 contain possess 902:908 arg2 pattern 924:930 a similar PCI pattern 910:930 a similar PCI pattern 910:930 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 5 4 contain possess 902:908 arg1 individuals 861:871 individuals 861:871 individuals of two different ethnicities 861:900 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 0 5 theme C 78:78 arg1 inhibitor 80:88 human blood protein C inhibitor 58:88 human blood protein C inhibitor 58:88 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 6 6 theme core 1114:1117 arg1 NeuAcGalGalNAc 1152:1165 NeuAcGalGalNAc 1152:1165 NeuAcGalGalNAc 1152:1165 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 6 theme core 1114:1117 arg1 O-glycan 1126:1133 a core type 1 O-glycan 1112:1133 a core type 1 O-glycan 1112:1133 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 7 with Thr20 1101:1105 arg1 NeuAcGalGalNAc 1152:1165 NeuAcGalGalNAc 1152:1165 NeuAcGalGalNAc 1152:1165 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 7 with Thr20 1101:1105 arg1 O-glycan 1126:1133 a core type 1 O-glycan 1112:1133 a core type 1 O-glycan 1112:1133 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 4 8 attach isolated 478:485 arg1 plasma 504:509 human blood plasma 492:509 human blood plasma 492:509 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 8 attach isolated 478:485 arg2 PCI 474:476 PCI 474:476 PCI isolated from human blood plasma 474:509 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 9 from amino-terminus 733:746 arg1 absence 701:707 absence 701:707 absence 701:707 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 9 from amino-terminus 733:746 arg1 presence 689:696 presence 689:696 presence 689:696 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 0 10 theme protein 70:76 arg1 inhibitor 80:88 human blood protein C inhibitor 58:88 human blood protein C inhibitor 58:88 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 2 11 theme superfamily 221:231 arg1 specificity 282:292 unusually broad protease specificity 257:292 unusually broad protease specificity 257:292 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 2 11 theme superfamily 221:231 arg1 member 200:205 a member 198:205 a member of the serpin superfamily of proteins 198:243 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 3 12 theme tumors 419:424 arg1 fertilization 390:402 fertilization 390:402 fertilization 390:402 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 12 theme tumors 419:424 arg1 coagulation 377:387 blood coagulation 371:387 blood coagulation 371:387 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 12 theme tumors 419:424 arg1 defence 439:445 pathogen defence 430:445 pathogen defence 430:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 12 theme tumors 419:424 arg1 prevention 405:414 prevention 405:414 prevention of tumors 405:424 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 2 13 theme serpin 214:219 arg1 superfamily 221:231 the serpin superfamily 210:231 the serpin superfamily of proteins 210:243 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 1 14 theme 57-kDa 122:127 arg1 inhibitor 101:109 Protein C inhibitor 91:109 Protein C inhibitor (PCI) 91:115 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 1 14 theme 57-kDa 122:127 arg1 glycoprotein 129:140 a 57-kDa glycoprotein 120:140 a 57-kDa glycoprotein that exists in many tissues and secretions in human 120:192 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 6 15 gly O-glycosylated 1083:1096 arg2 Thr20 1101:1105 Thr20 1101:1105 Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc 1101:1165 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 15 gly O-glycosylated 1083:1096 arg2 PCI 1056:1058 PCI 1056:1058 PCI in both individuals 1056:1078 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 15 gly O-glycosylated 1083:1096 arg1 PCI 1056:1058 PCI 1056:1058 PCI in both individuals 1056:1078 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 15 gly O-glycosylated 1083:1096 arg1 Thr20 1101:1105 Thr20 1101:1105 Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc 1101:1165 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 5 16 theme different 880:888 arg1 ethnicities 890:900 two different ethnicities 876:900 two different ethnicities 876:900 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 4 17 theme human 492:496 arg1 plasma 504:509 human blood plasma 492:509 human blood plasma 492:509 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 18 theme forms 664:668 arg1 occupancy 629:637 N-glycosylation occupancy 613:637 N-glycosylation occupancy 613:637 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 18 theme forms 664:668 arg1 presence 648:655 the presence 644:655 the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus 644:746 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 18 theme forms 664:668 arg1 structures 601:610 N-glycan structures 592:610 N-glycan structures 592:610 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 19 from differences 577:587 arg1 occupancy 629:637 N-glycosylation occupancy 613:637 N-glycosylation occupancy 613:637 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 19 from differences 577:587 arg1 presence 648:655 the presence 644:655 the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus 644:746 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 19 from differences 577:587 arg1 structures 601:610 N-glycan structures 592:610 N-glycan structures 592:610 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 0 20 theme Further 0:6 arg1 insight 8:14 Further insight 0:14 Further insight into the roles of the glycans 0:44 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 4 21 theme N-glycan 592:599 arg1 structures 601:610 N-glycan structures 592:610 N-glycan structures 592:610 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 7 22 theme several 1248:1254 arg1 sites 1271:1275 several ligand-binding sites 1248:1275 several ligand-binding sites of the inhibitor 1248:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 22 theme several 1248:1254 arg1 inhibitor 1284:1292 the inhibitor 1280:1292 the inhibitor 1280:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 5 23 theme single 832:837 arg1 individuals 839:849 single individuals 832:849 single individuals 832:849 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 6 24 from PCI 1056:1058 arg1 individuals 1068:1078 both individuals 1063:1078 both individuals 1063:1078 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 4 25 attach presence 689:696 arg2 acids 720:724 6 amino acids 712:724 6 amino acids 712:724 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 25 attach presence 689:696 arg1 amino-terminus 733:746 the amino-terminus 729:746 the amino-terminus 729:746 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 2 26 theme proteins 236:243 arg1 superfamily 221:231 the serpin superfamily 210:231 the serpin superfamily of proteins 210:243 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 1 27 theme many 157:160 arg1 tissues 162:168 many tissues 157:168 many tissues 157:168 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 3 28 theme pathogen 430:437 arg1 defence 439:445 pathogen defence 430:445 pathogen defence 430:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 5 29 theme PCI 920:922 arg1 pattern 924:930 a similar PCI pattern 910:930 a similar PCI pattern 910:930 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 7 30 theme O-glycan 1196:1203 arg1 site 1216:1219 the O-glycan attachment site 1192:1219 the O-glycan attachment site 1192:1219 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 30 theme O-glycan 1196:1203 arg1 located 1224:1230 located 1224:1230 located 1224:1230 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 3 31 theme blood 371:375 arg1 coagulation 377:387 blood coagulation 371:387 blood coagulation 371:387 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 32 theme processes 350:358 arg1 prevention 405:414 prevention 405:414 prevention of tumors 405:424 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 32 theme processes 350:358 arg1 range 341:345 a wide range 334:345 a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence 334:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 32 theme processes 350:358 arg1 coagulation 377:387 blood coagulation 371:387 blood coagulation 371:387 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 32 theme processes 350:358 arg1 defence 439:445 pathogen defence 430:445 pathogen defence 430:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 32 theme processes 350:358 arg1 fertilization 390:402 fertilization 390:402 fertilization 390:402 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 4 33 theme acids 720:724 arg1 absence 701:707 absence 701:707 absence 701:707 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 33 theme acids 720:724 arg1 presence 689:696 presence 689:696 presence 689:696 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 0 34 attach attached 46:53 arg1 inhibitor 80:88 human blood protein C inhibitor 58:88 human blood protein C inhibitor 58:88 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 0 34 attach attached 46:53 arg2 insight 8:14 Further insight 0:14 Further insight into the roles of the glycans 0:44 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 7 35 theme attachment 1205:1214 arg1 site 1216:1219 the O-glycan attachment site 1192:1219 the O-glycan attachment site 1192:1219 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 35 theme attachment 1205:1214 arg1 located 1224:1230 located 1224:1230 located 1224:1230 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 0 36 theme glycans 38:44 arg1 roles 25:29 the roles 21:29 the roles of the glycans 21:44 Further insight into the roles of the glycans attached to human blood protein C inhibitor. 21056543 5 37 theme such 785:788 arg1 heterogeneity 790:802 such heterogeneity 785:802 such heterogeneity 785:802 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 4 38 gly occupancy 629:637 arg2 forms 664:668 two forms 660:668 two forms that differ by the presence or absence of 6 amino acids at the amino-terminus 660:746 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 1 39 theme Protein 91:97 arg1 inhibitor 101:109 Protein C inhibitor 91:109 Protein C inhibitor (PCI) 91:115 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 1 39 theme Protein 91:97 arg1 glycoprotein 129:140 a 57-kDa glycoprotein 120:140 a 57-kDa glycoprotein that exists in many tissues and secretions in human 120:192 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 1 39 theme Protein 91:97 arg1 PCI 112:114 PCI 112:114 PCI 112:114 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 3 40 theme range 341:345 arg1 regulation 320:329 the regulation 316:329 the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence 316:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 7 41 theme inhibitor 1284:1292 arg1 sites 1271:1275 several ligand-binding sites 1248:1275 several ligand-binding sites of the inhibitor 1248:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 41 theme inhibitor 1284:1292 arg1 inhibitor 1284:1292 the inhibitor 1280:1292 the inhibitor 1280:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 1 42 theme C 99:99 arg1 inhibitor 101:109 Protein C inhibitor 91:109 Protein C inhibitor (PCI) 91:115 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 1 42 theme C 99:99 arg1 glycoprotein 129:140 a 57-kDa glycoprotein 120:140 a 57-kDa glycoprotein that exists in many tissues and secretions in human 120:192 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 1 42 theme C 99:99 arg1 PCI 112:114 PCI 112:114 PCI 112:114 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 7 43 theme ligand-binding 1256:1269 arg1 sites 1271:1275 several ligand-binding sites 1248:1275 several ligand-binding sites of the inhibitor 1248:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 43 theme ligand-binding 1256:1269 arg1 inhibitor 1284:1292 the inhibitor 1280:1292 the inhibitor 1280:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 1 44 from secretions 174:183 arg1 human 188:192 human 188:192 human 188:192 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 6 45 theme type 1119:1122 arg1 NeuAcGalGalNAc 1152:1165 NeuAcGalGalNAc 1152:1165 NeuAcGalGalNAc 1152:1165 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 6 45 theme type 1119:1122 arg1 O-glycan 1126:1133 a core type 1 O-glycan 1112:1133 a core type 1 O-glycan 1112:1133 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 5 46 theme ethnicities 890:900 arg1 individuals 861:871 individuals 861:871 individuals of two different ethnicities 861:900 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 2 47 theme protease 273:280 arg1 specificity 282:292 unusually broad protease specificity 257:292 unusually broad protease specificity 257:292 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 2 47 theme protease 273:280 arg1 member 200:205 a member 198:205 a member of the serpin superfamily of proteins 198:243 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 1 48 from tissues 162:168 arg1 human 188:192 human 188:192 human 188:192 Protein C inhibitor (PCI) is a 57-kDa glycoprotein that exists in many tissues and secretions in human. 21056543 7 49 from located 1224:1230 arg1 proximity 1235:1243 proximity 1235:1243 proximity to several ligand-binding sites of the inhibitor 1235:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 2 50 theme broad 267:271 arg1 specificity 282:292 unusually broad protease specificity 257:292 unusually broad protease specificity 257:292 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 2 50 theme broad 267:271 arg1 member 200:205 a member 198:205 a member of the serpin superfamily of proteins 198:243 As a member of the serpin superfamily of proteins it displays unusually broad protease specificity. 21056543 6 51 theme experimental 1029:1040 arg1 evidence 1042:1049 experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc 1029:1165 experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc 1029:1165 Furthermore, we have provided experimental evidence that PCI in both individuals is O-glycosylated on Thr20 with a core type 1 O-glycan, which is mostly NeuAcGalGalNAc. 21056543 7 52 located located 1224:1230 arg1 proximity 1235:1243 proximity 1235:1243 proximity to several ligand-binding sites of the inhibitor 1235:1292 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 52 located located 1224:1230 arg2 located 1224:1230 located 1224:1230 located 1224:1230 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 52 located located 1224:1230 arg2 site 1216:1219 the O-glycan attachment site 1192:1219 the O-glycan attachment site 1192:1219 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 4 53 theme amino 714:718 arg1 acids 720:724 6 amino acids 712:724 6 amino acids 712:724 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 4 54 theme N-glycosylation 613:627 arg1 occupancy 629:637 N-glycosylation occupancy 613:637 N-glycosylation occupancy 613:637 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 21056543 3 55 theme wide 336:339 arg1 prevention 405:414 prevention 405:414 prevention of tumors 405:424 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 55 theme wide 336:339 arg1 range 341:345 a wide range 334:345 a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence 334:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 55 theme wide 336:339 arg1 coagulation 377:387 blood coagulation 371:387 blood coagulation 371:387 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 55 theme wide 336:339 arg1 defence 439:445 pathogen defence 430:445 pathogen defence 430:445 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 3 55 theme wide 336:339 arg1 fertilization 390:402 fertilization 390:402 fertilization 390:402 PCI is implicated in the regulation of a wide range of processes, including blood coagulation, fertilization, prevention of tumors and pathogen defence. 21056543 5 56 theme similar 912:918 arg1 pattern 924:930 a similar PCI pattern 910:930 a similar PCI pattern 910:930 In this study we have verified that such heterogeneity exists in PCI purified from single individuals, and that individuals of two different ethnicities possess a similar PCI pattern, verifying that the micro-heterogeneity is conserved among humans. 21056543 7 57 from proximity 1235:1243 arg1 site 1216:1219 the O-glycan attachment site 1192:1219 the O-glycan attachment site 1192:1219 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 7 57 from proximity 1235:1243 arg1 located 1224:1230 located 1224:1230 located 1224:1230 Modeling suggested that the O-glycan attachment site is located in proximity to several ligand-binding sites of the inhibitor. 21056543 4 58 dep presence 689:696 arg1 the 685:687 the 685:687 the 685:687 It has been reported that PCI isolated from human blood plasma is highly heterogeneous, and that this heterogeneity is caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of two forms that differ by the presence or absence of 6 amino acids at the amino-terminus. 23527852 2 0 contain containing 279:288 arg2 O-glycosylation 310:324 a mucin-type core-1 O-glycosylation 290:324 a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids 290:366 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 2 0 contain containing 279:288 arg1 all 275:277 all 275:277 all 275:277 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 4 1 theme apoCIII 600:606 arg1 isoforms 608:615 apoCIII isoforms 600:615 apoCIII isoforms in serum protein profiles 600:641 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 6 2 theme new 816:818 arg1 isoforms 828:835 Six new apoCIII isoforms 812:835 Six new apoCIII isoforms 812:835 Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. 23527852 2 3 theme core-1 303:308 arg1 O-glycosylation 310:324 a mucin-type core-1 O-glycosylation 290:324 a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids 290:366 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 9 4 gly fucosylated 1205:1215 arg1 isoforms 1217:1224 the fucosylated isoforms 1201:1224 the fucosylated isoforms 1201:1224 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 6 5 theme ultrahigh 894:902 arg1 precision 904:912 ultrahigh precision 894:912 ultrahigh precision 894:912 Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. 23527852 10 6 dep implications 1367:1378 arg1 have 1401:1404 have 1401:1404 may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies 1397:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 3 7 from Changes 369:375 arg1 abundance 393:401 the relative abundance 380:401 the relative abundance of these glycoforms 380:421 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 5 8 theme RPC18-based 720:730 arg1 method 746:751 a fully automated RPC18-based magnetic bead method 702:751 a fully automated RPC18-based magnetic bead method 702:751 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 9 9 theme serum 1251:1255 arg1 samples 1257:1263 96 serum samples 1248:1263 96 serum samples 1248:1263 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 1 10 gly glycoprotein 175:186 arg1 Apolipoprotein-CIII 124:142 Apolipoprotein-CIII 124:142 Apolipoprotein-CIII (apoCIII) 124:152 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 1 10 gly glycoprotein 175:186 arg1 glycoprotein 175:186 an abundant blood glycoprotein 157:186 an abundant blood glycoprotein associated with lipoprotein particles 157:224 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 10 11 gly fucosylated 1284:1294 arg1 isoforms 1316:1323 fucosylated apolipoprotein-CIII isoforms 1284:1323 fucosylated apolipoprotein-CIII isoforms 1284:1323 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 9 12 theme fucosylated 1205:1215 arg1 isoforms 1217:1224 the fucosylated isoforms 1201:1224 the fucosylated isoforms 1201:1224 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 7 13 gly fucosylated 981:991 arg1 moieties 963:970 more complex glycan moieties 943:970 more complex glycan moieties that are fucosylated instead of sialylated 943:1013 These were characterized by more complex glycan moieties that are fucosylated instead of sialylated. 23527852 1 14 theme abundant 160:167 arg1 Apolipoprotein-CIII 124:142 Apolipoprotein-CIII 124:142 Apolipoprotein-CIII (apoCIII) 124:152 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 1 14 theme abundant 160:167 arg1 glycoprotein 175:186 an abundant blood glycoprotein 157:186 an abundant blood glycoprotein associated with lipoprotein particles 157:224 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 8 15 theme trap 1128:1131 arg1 MS/MS 1133:1137 bottom-up LC-ion trap MS/MS 1111:1137 bottom-up LC-ion trap MS/MS 1111:1137 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 4 16 theme 15T 517:519 arg1 Fourier 527:533 ultrahigh resolution 15T MALDI Fourier 496:533 ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS 496:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 4 17 from isoforms 608:615 arg1 profiles 634:641 serum protein profiles 620:641 serum protein profiles 620:641 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 9 18 theme isoforms 1217:1224 arg1 abundance 1188:1196 abundance 1188:1196 abundance 1188:1196 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 9 18 theme isoforms 1217:1224 arg1 presence 1175:1182 presence 1175:1182 presence 1175:1182 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 0 19 theme human 112:116 arg1 sera 118:121 human sera 112:121 human sera 112:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 8 20 theme glycosylation 1062:1074 arg1 site 1076:1079 the glycosylation site 1058:1079 the glycosylation site 1058:1079 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 10 21 theme different 1507:1515 arg1 pathologies 1517:1527 many different pathologies 1502:1527 many different pathologies 1502:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 10 22 theme isoforms 1316:1323 arg1 findings 1272:1279 These findings 1266:1279 These findings of fucosylated apolipoprotein-CIII isoforms 1266:1323 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 4 23 theme ultrahigh 496:504 arg1 Fourier 527:533 ultrahigh resolution 15T MALDI Fourier 496:533 ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS 496:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 10 24 theme fucosylated 1284:1294 arg1 isoforms 1316:1323 fucosylated apolipoprotein-CIII isoforms 1284:1323 fucosylated apolipoprotein-CIII isoforms 1284:1323 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 7 25 theme complex 948:954 arg1 moieties 963:970 more complex glycan moieties 943:970 more complex glycan moieties that are fucosylated instead of sialylated 943:1013 These were characterized by more complex glycan moieties that are fucosylated instead of sialylated. 23527852 0 26 theme new 18:20 arg1 glycoforms 42:51 new apolipoprotein-CIII glycoforms 18:51 new apolipoprotein-CIII glycoforms 18:51 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 8 27 theme bottom-up 1111:1119 arg1 MS/MS 1133:1137 bottom-up LC-ion trap MS/MS 1111:1137 bottom-up LC-ion trap MS/MS 1111:1137 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 9 28 dep presence 1175:1182 arg1 the 1171:1173 the 1171:1173 the 1171:1173 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 0 29 theme MALDI-FTICR 79:89 arg1 spectrometry 96:107 ultrahigh resolution MALDI-FTICR mass spectrometry 58:107 ultrahigh resolution MALDI-FTICR mass spectrometry of human sera 58:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 10 30 theme further 1333:1339 arg1 research 1341:1348 further research 1333:1348 further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies 1333:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 3 31 theme pathologies 468:478 arg1 variety 447:453 a variety 445:453 a variety of different pathologies 445:478 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 3 31 theme pathologies 468:478 arg1 pathologies 468:478 different pathologies 458:478 different pathologies 458:478 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 0 32 theme glycoforms 42:51 arg1 Identification 0:13 Identification 0:13 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 0:122 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 5 33 theme extraction 793:802 arg1 method 804:809 an RPC4 cartridge-based solid phase extraction method 757:809 an RPC4 cartridge-based solid phase extraction method 757:809 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 6 34 theme measurement 871:881 arg1 errors 883:888 low-ppm mass measurement errors 858:888 low-ppm mass measurement errors 858:888 Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. 23527852 4 35 theme ion 545:547 arg1 resonance 559:567 ion cyclotron resonance 545:567 ion cyclotron resonance (FTICR) MS 545:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 4 35 theme ion 545:547 arg1 FTICR 570:574 FTICR 570:574 FTICR 570:574 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 5 36 theme RPC4 760:763 arg1 method 804:809 an RPC4 cartridge-based solid phase extraction method 757:809 an RPC4 cartridge-based solid phase extraction method 757:809 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 0 37 theme ultrahigh 58:66 arg1 spectrometry 96:107 ultrahigh resolution MALDI-FTICR mass spectrometry 58:107 ultrahigh resolution MALDI-FTICR mass spectrometry of human sera 58:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 6 38 theme low-ppm 858:864 arg1 errors 883:888 low-ppm mass measurement errors 858:888 low-ppm mass measurement errors 858:888 Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. 23527852 4 39 theme resonance 559:567 arg1 MS 577:578 ion cyclotron resonance (FTICR) MS 545:578 ion cyclotron resonance (FTICR) MS 545:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 2 40 theme different 233:241 arg1 glycoforms 243:252 Three different glycoforms 227:252 Three different glycoforms 227:252 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 0 41 with Identification 0:13 arg1 spectrometry 96:107 ultrahigh resolution MALDI-FTICR mass spectrometry 58:107 ultrahigh resolution MALDI-FTICR mass spectrometry of human sera 58:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 2 42 theme sialic 355:360 arg1 acids 362:366 either zero, one or two sialic acids 331:366 either zero, one or two sialic acids 331:366 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 10 43 from alterations 1440:1450 arg1 apoCIII 1455:1461 apoCIII 1455:1461 apoCIII 1455:1461 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 4 44 theme serum 620:624 arg1 profiles 634:641 serum protein profiles 620:641 serum protein profiles 620:641 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 5 45 theme serum 662:666 arg1 proteins 668:675 serum proteins 662:675 serum proteins 662:675 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 6 46 theme apoCIII 820:826 arg1 isoforms 828:835 Six new apoCIII isoforms 812:835 Six new apoCIII isoforms 812:835 Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. 23527852 8 47 theme glycan 1031:1036 arg1 moiety 1038:1043 the glycan moiety 1027:1043 the glycan moiety 1027:1043 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 5 48 theme solid 781:785 arg1 method 804:809 an RPC4 cartridge-based solid phase extraction method 757:809 an RPC4 cartridge-based solid phase extraction method 757:809 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 1 49 theme lipoprotein 204:214 arg1 particles 216:224 lipoprotein particles 204:224 lipoprotein particles 204:224 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 5 50 theme automated 710:718 arg1 method 746:751 a fully automated RPC18-based magnetic bead method 702:751 a fully automated RPC18-based magnetic bead method 702:751 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 9 51 from variation 1158:1166 arg1 abundance 1188:1196 abundance 1188:1196 abundance 1188:1196 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 9 51 from variation 1158:1166 arg1 presence 1175:1182 presence 1175:1182 presence 1175:1182 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 5 52 theme magnetic 732:739 arg1 method 746:751 a fully automated RPC18-based magnetic bead method 702:751 a fully automated RPC18-based magnetic bead method 702:751 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 9 53 theme samples 1257:1263 arg1 samples 1257:1263 96 serum samples 1248:1263 96 serum samples 1248:1263 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 9 53 theme samples 1257:1263 arg1 set 1241:1243 a set 1239:1243 a set of 96 serum samples 1239:1263 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 10 54 theme studies 1426:1432 arg1 plethora 1414:1421 the plethora 1410:1421 the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies 1410:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 4 55 used used 584:587 arg2 Fourier 527:533 ultrahigh resolution 15T MALDI Fourier 496:533 ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS 496:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 2 56 theme mucin-type 292:301 arg1 O-glycosylation 310:324 a mucin-type core-1 O-glycosylation 290:324 a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids 290:366 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 2 57 with O-glycosylation 310:324 arg1 acids 362:366 either zero, one or two sialic acids 331:366 either zero, one or two sialic acids 331:366 Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. 23527852 8 58 gly glycosylation 1062:1074 arg2 site 1076:1079 the glycosylation site 1058:1079 the glycosylation site 1058:1079 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 0 59 theme mass 91:94 arg1 spectrometry 96:107 ultrahigh resolution MALDI-FTICR mass spectrometry 58:107 ultrahigh resolution MALDI-FTICR mass spectrometry of human sera 58:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 4 60 theme MALDI 521:525 arg1 Fourier 527:533 ultrahigh resolution 15T MALDI Fourier 496:533 ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS 496:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 5 61 theme bead 741:744 arg1 method 746:751 a fully automated RPC18-based magnetic bead method 702:751 a fully automated RPC18-based magnetic bead method 702:751 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 5 62 theme cartridge-based 765:779 arg1 method 804:809 an RPC4 cartridge-based solid phase extraction method 757:809 an RPC4 cartridge-based solid phase extraction method 757:809 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 1 63 theme blood 169:173 arg1 Apolipoprotein-CIII 124:142 Apolipoprotein-CIII 124:142 Apolipoprotein-CIII (apoCIII) 124:152 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 1 63 theme blood 169:173 arg1 glycoprotein 175:186 an abundant blood glycoprotein 157:186 an abundant blood glycoprotein associated with lipoprotein particles 157:224 Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. 23527852 9 64 located found 1230:1234 arg1 samples 1257:1263 96 serum samples 1248:1263 96 serum samples 1248:1263 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 9 64 located found 1230:1234 arg2 variation 1158:1166 A large variation 1150:1166 A large variation in the presence and abundance of the fucosylated isoforms 1150:1224 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 9 64 located found 1230:1234 arg1 set 1241:1243 a set 1239:1243 a set of 96 serum samples 1239:1263 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 0 65 theme sera 118:121 arg1 spectrometry 96:107 ultrahigh resolution MALDI-FTICR mass spectrometry 58:107 ultrahigh resolution MALDI-FTICR mass spectrometry of human sera 58:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 3 66 theme relative 384:391 arg1 abundance 393:401 the relative abundance 380:401 the relative abundance of these glycoforms 380:421 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 8 67 theme top-down 1082:1089 arg1 ESI-FTICR-MS/MS 1091:1105 top-down ESI-FTICR-MS/MS 1082:1105 top-down ESI-FTICR-MS/MS 1082:1105 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 4 68 theme resolution 506:515 arg1 Fourier 527:533 ultrahigh resolution 15T MALDI Fourier 496:533 ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS 496:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 10 69 theme many 1502:1505 arg1 pathologies 1517:1527 many different pathologies 1502:1527 many different pathologies 1502:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 10 70 theme apolipoprotein-CIII 1296:1314 arg1 isoforms 1316:1323 fucosylated apolipoprotein-CIII isoforms 1284:1323 fucosylated apolipoprotein-CIII isoforms 1284:1323 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 3 71 theme different 458:466 arg1 pathologies 468:478 different pathologies 458:478 different pathologies 458:478 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 3 72 theme glycoforms 412:421 arg1 abundance 393:401 the relative abundance 380:401 the relative abundance of these glycoforms 380:421 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 0 73 theme apolipoprotein-CIII 22:40 arg1 glycoforms 42:51 new apolipoprotein-CIII glycoforms 18:51 new apolipoprotein-CIII glycoforms 18:51 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 8 74 theme LC-ion 1121:1126 arg1 MS/MS 1133:1137 bottom-up LC-ion trap MS/MS 1111:1137 bottom-up LC-ion trap MS/MS 1111:1137 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 7 75 theme glycan 956:961 arg1 moieties 963:970 more complex glycan moieties 943:970 more complex glycan moieties that are fucosylated instead of sialylated 943:1013 These were characterized by more complex glycan moieties that are fucosylated instead of sialylated. 23527852 5 76 theme phase 787:791 arg1 method 804:809 an RPC4 cartridge-based solid phase extraction method 757:809 an RPC4 cartridge-based solid phase extraction method 757:809 For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. 23527852 3 77 located observed 433:440 arg1 variety 447:453 a variety 445:453 a variety of different pathologies 445:478 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 3 77 located observed 433:440 arg1 pathologies 468:478 different pathologies 458:478 different pathologies 458:478 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 3 77 located observed 433:440 arg2 Changes 369:375 Changes 369:375 Changes in the relative abundance of these glycoforms 369:421 Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. 23527852 4 78 theme cyclotron 549:557 arg1 resonance 559:567 ion cyclotron resonance 545:567 ion cyclotron resonance (FTICR) MS 545:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 4 78 theme cyclotron 549:557 arg1 FTICR 570:574 FTICR 570:574 FTICR 570:574 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 10 79 theme pathologies 1517:1527 arg1 development 1487:1497 the development 1483:1497 the development of many different pathologies 1483:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 9 80 theme large 1152:1156 arg1 variation 1158:1166 A large variation 1150:1166 A large variation in the presence and abundance of the fucosylated isoforms 1150:1224 A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. 23527852 10 81 attach linked 1473:1478 arg2 alterations 1440:1450 alterations 1440:1450 alterations in apoCIII 1440:1461 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 10 81 attach linked 1473:1478 arg1 development 1487:1497 the development 1483:1497 the development of many different pathologies 1483:1527 These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies. 23527852 0 82 theme resolution 68:77 arg1 spectrometry 96:107 ultrahigh resolution MALDI-FTICR mass spectrometry 58:107 ultrahigh resolution MALDI-FTICR mass spectrometry of human sera 58:121 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 4 83 dep Fourier 527:533 arg1 transform 535:543 transform 535:543 transform ion cyclotron resonance (FTICR) MS 535:578 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 23527852 6 84 theme mass 866:869 arg1 errors 883:888 low-ppm mass measurement errors 858:888 low-ppm mass measurement errors 858:888 Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. 23527852 8 85 used used 1144:1147 arg2 ESI-FTICR-MS/MS 1091:1105 top-down ESI-FTICR-MS/MS 1082:1105 top-down ESI-FTICR-MS/MS 1082:1105 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 8 85 used used 1144:1147 arg2 MS/MS 1133:1137 bottom-up LC-ion trap MS/MS 1111:1137 bottom-up LC-ion trap MS/MS 1111:1137 To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. 23527852 0 86 gly glycoforms 42:51 arg1 apolipoprotein-CIII 22:40 new apolipoprotein-CIII glycoforms 18:51 new apolipoprotein-CIII glycoforms 18:51 Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera. 23527852 4 87 theme protein 626:632 arg1 profiles 634:641 serum protein profiles 620:641 serum protein profiles 620:641 In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. 15863501 0 0 theme group 91:95 arg1 potential 72:80 tumorigenic potential 60:80 tumorigenic potential 60:80 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 0 0 theme group 91:95 arg1 distribution 9:20 Cellular distribution 0:20 Cellular distribution 0:20 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 0 0 theme group 91:95 arg1 modification 42:53 post-translational modification 23:53 post-translational modification 23:53 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 1 1 theme unique 246:251 arg1 N- 253:254 unique N- 246:254 unique N- 246:254 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 9 2 theme cancer 1676:1681 arg1 development 1683:1693 cancer development 1676:1693 cancer development 1676:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 3 theme type-specific 1575:1587 arg1 processing 1589:1598 unique cell type-specific processing 1563:1598 unique cell type-specific processing 1563:1598 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 4 dep cell 1716:1719 arg1 angiogenesis 1732:1743 angiogenesis 1732:1743 angiogenesis 1732:1743 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 4 dep cell 1716:1719 arg1 growth 1721:1726 growth 1721:1726 growth 1721:1726 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 0 5 theme human 85:89 arg1 group 91:95 human group III 85:99 human group III 85:99 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 6 6 theme sPLA 1020:1023 arg1 Expression 1006:1015 Expression 1006:1015 Expression of sPLA(2)-III 1006:1030 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 3 7 gly N-glycosylated 651:664 arg2 sites 683:687 two consensus sites 669:687 two consensus sites 669:687 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 7 gly N-glycosylated 651:664 arg1 production 569:578 the production 565:578 the production 565:578 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 7 gly N-glycosylated 651:664 arg1 N-glycosylated 651:664 N-glycosylated 651:664 N-glycosylated 651:664 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 1 8 theme secreted 146:153 arg1 2 171:171 2 171:171 2 171:171 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 8 theme secreted 146:153 arg1 sPLA 175:178 sPLA(2)-III 175:185 sPLA(2)-III 175:185 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 8 theme secreted 146:153 arg1 A 169:169 Human group III secreted phospholipase A 130:169 Human group III secreted phospholipase A(2) (sPLA(2)-III) 130:186 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 6 9 theme cell 1176:1179 arg1 proliferation 1181:1193 cell proliferation 1176:1193 cell proliferation 1176:1193 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 3 10 dep N- 483:484 arg1 the 479:481 the 479:481 the 479:481 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 9 11 from N-glycosylation 1604:1618 arg1 development 1683:1693 cancer development 1676:1693 cancer development 1676:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 2 12 theme cell 433:436 arg1 types 438:442 various cell types 425:442 various cell types 425:442 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 4 13 with tissues 812:818 arg1 inflammation 825:836 inflammation 825:836 inflammation 825:836 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 4 13 with tissues 812:818 arg1 injury 848:853 ischemic injury 839:853 ischemic injury 839:853 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 4 13 with tissues 812:818 arg1 cancer 860:865 cancer 860:865 cancer 860:865 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 4 14 theme ischemic 839:846 arg1 injury 848:853 ischemic injury 839:853 ischemic injury 839:853 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 9 15 theme potential 1643:1651 arg1 role 1653:1656 the potential role 1639:1656 the potential role of this enzyme in cancer development 1639:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 1 16 theme C-terminal 260:269 arg1 domains 271:277 C-terminal domains 260:277 C-terminal domains 260:277 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 2 17 theme various 425:431 arg1 types 438:442 various cell types 425:442 various cell types 425:442 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 2 18 from functions 412:420 arg1 types 438:442 various cell types 425:442 various cell types 425:442 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 3 19 theme consensus 673:681 arg1 sites 683:687 two consensus sites 669:687 two consensus sites 669:687 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 1 20 theme central 202:208 arg1 domain 228:233 a central group III sPLA(2) domain 200:233 a central group III sPLA(2) domain flanked by unique N- and C-terminal domains 200:277 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 8 21 theme small 1402:1406 arg1 RNA 1420:1422 small interfering RNA 1402:1422 small interfering RNA for sPLA(2)-III 1402:1438 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 7 22 theme larger 1315:1320 arg1 tumors 1328:1333 larger solid tumors 1315:1333 larger solid tumors 1315:1333 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 1 23 theme phospholipase 155:167 arg1 2 171:171 2 171:171 2 171:171 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 23 theme phospholipase 155:167 arg1 sPLA 175:178 sPLA(2)-III 175:185 sPLA(2)-III 175:185 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 23 theme phospholipase 155:167 arg1 A 169:169 Human group III secreted phospholipase A 130:169 Human group III secreted phospholipase A(2) (sPLA(2)-III) 130:186 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 0 24 theme phospholipase 110:122 arg1 2 126:126 2 126:126 2 126:126 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 0 24 theme phospholipase 110:122 arg1 A 124:124 phospholipase A 110:124 phospholipase A(2) 110:127 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 8 25 theme cancer 1512:1517 arg1 cells 1519:1523 colorectal cancer cells 1501:1523 colorectal cancer cells 1501:1523 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 7 26 theme control 1377:1383 arg1 cells 1385:1389 control cells 1377:1389 control cells 1377:1389 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 4 27 theme human 806:810 arg1 tissues 812:818 human tissues 806:818 human tissues with inflammation, ischemic injury, and cancer 806:865 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 5 28 theme this 882:885 arg1 support 871:877 support 871:877 support of this 871:885 In support of this, sPLA(2)-III was induced in cultured microvascular endothelial cells after stimulation with proinflammatory cytokines. 15863501 5 29 theme microvascular 924:936 arg1 cells 950:954 cultured microvascular endothelial cells 915:954 cultured microvascular endothelial cells 915:954 In support of this, sPLA(2)-III was induced in cultured microvascular endothelial cells after stimulation with proinflammatory cytokines. 15863501 8 30 theme colorectal 1501:1510 arg1 cells 1519:1523 colorectal cancer cells 1501:1523 colorectal cancer cells 1501:1523 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 8 31 theme interfering 1408:1418 arg1 RNA 1420:1422 small interfering RNA 1402:1422 small interfering RNA for sPLA(2)-III 1402:1438 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 0 32 theme Cellular 0:7 arg1 distribution 9:20 Cellular distribution 0:20 Cellular distribution 0:20 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 1 33 theme group 210:214 arg1 domain 228:233 a central group III sPLA(2) domain 200:233 a central group III sPLA(2) domain flanked by unique N- and C-terminal domains 200:277 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 7 34 theme -III-transfected 1285:1300 arg1 cells 1302:1306 the sPLA(2)-III-transfected cells 1274:1306 the sPLA(2)-III-transfected cells 1274:1306 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 6 35 theme catalytic 1223:1231 arg1 activity 1233:1240 sPLA(2)-III catalytic activity 1211:1240 sPLA(2)-III catalytic activity 1211:1240 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 9 36 from role 1653:1656 arg1 development 1683:1693 cancer development 1676:1693 cancer development 1676:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 0 37 theme post-translational 23:40 arg1 modification 42:53 post-translational modification 23:53 post-translational modification 23:53 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 5 38 theme cultured 915:922 arg1 cells 950:954 cultured microvascular endothelial cells 915:954 cultured microvascular endothelial cells 915:954 In support of this, sPLA(2)-III was induced in cultured microvascular endothelial cells after stimulation with proinflammatory cytokines. 15863501 6 39 theme colorectal 1082:1091 arg1 cells 1100:1104 colorectal cancer cells 1082:1104 colorectal cancer cells transfected with sPLA(2)-III 1082:1133 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 7 40 theme nude 1263:1266 arg1 mice 1268:1271 nude mice 1263:1271 nude mice 1263:1271 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 3 41 theme sPLA 612:615 arg1 domain 620:625 only the sPLA(2) domain 603:625 only the sPLA(2) domain 603:625 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 9 42 theme enzyme 1666:1671 arg1 role 1653:1656 the potential role 1639:1656 the potential role of this enzyme in cancer development 1639:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 42 theme enzyme 1666:1671 arg1 N-glycosylation 1604:1618 N-glycosylation 1604:1618 N-glycosylation 1604:1618 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 42 theme enzyme 1666:1671 arg1 processing 1589:1598 unique cell type-specific processing 1563:1598 unique cell type-specific processing 1563:1598 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 8 43 theme PGE 1462:1464 arg1 production 1469:1478 PGE(2) production 1462:1478 PGE(2) production 1462:1478 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 1 44 theme sPLA 220:223 arg1 domain 228:233 a central group III sPLA(2) domain 200:233 a central group III sPLA(2) domain flanked by unique N- and C-terminal domains 200:277 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 7 45 theme increased 1340:1348 arg1 angiogenesis 1350:1361 increased angiogenesis 1340:1361 increased angiogenesis 1340:1361 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 6 46 theme PGE 1154:1156 arg1 production 1161:1170 enhanced PGE(2) production 1145:1170 enhanced PGE(2) production 1145:1170 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 5 47 theme endothelial 938:948 arg1 cells 950:954 cultured microvascular endothelial cells 915:954 cultured microvascular endothelial cells 915:954 In support of this, sPLA(2)-III was induced in cultured microvascular endothelial cells after stimulation with proinflammatory cytokines. 15863501 9 48 gly N-glycosylation 1604:1618 arg1 sPLA 1623:1626 sPLA(2)-III 1623:1633 sPLA(2)-III 1623:1633 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 48 gly N-glycosylation 1604:1618 arg1 enzyme 1666:1671 this enzyme 1661:1671 this enzyme 1661:1671 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 48 gly N-glycosylation 1604:1618 arg1 development 1683:1693 cancer development 1676:1693 cancer development 1676:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 49 theme unique 1563:1568 arg1 processing 1589:1598 unique cell type-specific processing 1563:1598 unique cell type-specific processing 1563:1598 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 2 50 theme sPLA 298:301 arg1 sufficient 323:332 sufficient 323:332 sufficient 323:332 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 2 50 theme sPLA 298:301 arg1 domain 306:311 the sPLA(2) domain 294:311 the sPLA(2) domain alone 294:317 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 4 51 from endothelium 791:801 arg1 tissues 812:818 human tissues 806:818 human tissues with inflammation, ischemic injury, and cancer 806:865 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 6 52 theme enhanced 1145:1152 arg1 production 1161:1170 enhanced PGE(2) production 1145:1170 enhanced PGE(2) production 1145:1170 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 5 53 with stimulation 962:972 arg1 cytokines 995:1003 proinflammatory cytokines 979:1003 proinflammatory cytokines 979:1003 In support of this, sPLA(2)-III was induced in cultured microvascular endothelial cells after stimulation with proinflammatory cytokines. 15863501 6 54 theme cancer 1093:1098 arg1 cells 1100:1104 colorectal cancer cells 1082:1104 colorectal cancer cells transfected with sPLA(2)-III 1082:1133 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 0 55 theme tumorigenic 60:70 arg1 potential 72:80 tumorigenic potential 60:80 tumorigenic potential 60:80 Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A(2). 15863501 6 56 theme tumor 1065:1069 arg1 cells 1071:1075 various tumor cells 1057:1075 various tumor cells 1057:1075 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 2 57 theme prostaglandin 373:385 arg1 2 389:389 2 389:389 2 389:389 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 2 57 theme prostaglandin 373:385 arg1 PGE 393:395 PGE(2) 393:398 PGE(2) 393:398 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 2 57 theme prostaglandin 373:385 arg1 E 387:387 its prostaglandin E 369:387 its prostaglandin E(2) (PGE(2))-generating functions in various cell types 369:442 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 4 58 theme microvascular 777:789 arg1 endothelium 791:801 the microvascular endothelium 773:801 the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer 773:865 Immunohistochemistry demonstrated that sPLA(2)-III was preferentially expressed in the microvascular endothelium in human tissues with inflammation, ischemic injury, and cancer. 15863501 6 59 theme various 1057:1063 arg1 cells 1071:1075 various tumor cells 1057:1075 various tumor cells 1057:1075 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 3 60 theme cell 467:470 arg1 types 472:476 not all cell types 459:476 not all cell types 459:476 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 61 from sites 683:687 arg1 production 569:578 the production 565:578 the production 565:578 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 61 from sites 683:687 arg1 N-glycosylated 651:664 N-glycosylated 651:664 N-glycosylated 651:664 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 2 62 theme -generating 400:410 arg1 functions 412:420 its prostaglandin E(2) (PGE(2))-generating functions 369:420 its prostaglandin E(2) (PGE(2))-generating functions in various cell types 369:442 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 3 63 theme C-terminal 490:499 arg1 sPLA 512:515 sPLA(2)-III 512:522 sPLA(2)-III 512:522 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 63 theme C-terminal 490:499 arg1 domains 501:507 C-terminal domains 490:507 C-terminal domains 490:507 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 9 64 from processing 1589:1598 arg1 development 1683:1693 cancer development 1676:1693 cancer development 1676:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 6 65 theme sPLA 1211:1214 arg1 activity 1233:1240 sPLA(2)-III catalytic activity 1211:1240 sPLA(2)-III catalytic activity 1211:1240 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 5 66 theme proinflammatory 979:993 arg1 cytokines 995:1003 proinflammatory cytokines 979:1003 proinflammatory cytokines 979:1003 In support of this, sPLA(2)-III was induced in cultured microvascular endothelial cells after stimulation with proinflammatory cytokines. 15863501 9 67 theme tumor 1710:1714 arg1 cell 1716:1719 tumor cell growth and angiogenesis 1710:1743 tumor cell growth and angiogenesis 1710:1743 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 8 68 theme cells 1519:1523 arg1 proliferation 1484:1496 proliferation 1484:1496 proliferation of colorectal cancer cells 1484:1523 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 8 68 theme cells 1519:1523 arg1 production 1469:1478 PGE(2) production 1462:1478 PGE(2) production 1462:1478 Moreover, small interfering RNA for sPLA(2)-III significantly reduced PGE(2) production and proliferation of colorectal cancer cells. 15863501 7 69 theme solid 1322:1326 arg1 tumors 1328:1333 larger solid tumors 1315:1333 larger solid tumors 1315:1333 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 9 70 theme sPLA 1623:1626 arg1 role 1653:1656 the potential role 1639:1656 the potential role of this enzyme in cancer development 1639:1693 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 70 theme sPLA 1623:1626 arg1 N-glycosylation 1604:1618 N-glycosylation 1604:1618 N-glycosylation 1604:1618 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 9 70 theme sPLA 1623:1626 arg1 processing 1589:1598 unique cell type-specific processing 1563:1598 unique cell type-specific processing 1563:1598 Taken together, these results reveal unique cell type-specific processing and N-glycosylation of sPLA(2)-III and the potential role of this enzyme in cancer development by stimulating tumor cell growth and angiogenesis. 15863501 2 71 theme E 387:387 arg1 functions 412:420 its prostaglandin E(2) (PGE(2))-generating functions 369:420 its prostaglandin E(2) (PGE(2))-generating functions in various cell types 369:442 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 6 72 theme sPLA 1123:1126 arg1 -III 1130:1133 sPLA(2)-III 1123:1133 sPLA(2)-III 1123:1133 Expression of sPLA(2)-III was also associated with various tumor cells, and colorectal cancer cells transfected with sPLA(2)-III exhibited enhanced PGE(2) production and cell proliferation, which required sPLA(2)-III catalytic activity. 15863501 7 73 theme sPLA 1278:1281 arg1 cells 1302:1306 the sPLA(2)-III-transfected cells 1274:1306 the sPLA(2)-III-transfected cells 1274:1306 When implanted into nude mice, the sPLA(2)-III-transfected cells formed larger solid tumors with increased angiogenesis compared with control cells. 15863501 1 74 theme Human 130:134 arg1 2 171:171 2 171:171 2 171:171 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 74 theme Human 130:134 arg1 sPLA 175:178 sPLA(2)-III 175:185 sPLA(2)-III 175:185 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 74 theme Human 130:134 arg1 A 169:169 Human group III secreted phospholipase A 130:169 Human group III secreted phospholipase A(2) (sPLA(2)-III) 130:186 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 2 75 theme catalytic 342:350 arg1 activity 352:359 its catalytic activity 338:359 its catalytic activity 338:359 We found that the sPLA(2) domain alone was sufficient for its catalytic activity and for its prostaglandin E(2) (PGE(2))-generating functions in various cell types. 15863501 3 76 theme sPLA 512:515 arg1 sPLA 512:515 sPLA(2)-III 512:522 sPLA(2)-III 512:522 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 76 theme sPLA 512:515 arg1 N- 483:484 N- 483:484 N- 483:484 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 76 theme sPLA 512:515 arg1 domains 501:507 C-terminal domains 490:507 C-terminal domains 490:507 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 77 contain containing 592:601 arg1 form 587:590 the form 583:590 the form containing only the sPLA(2) domain 583:625 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 77 contain containing 592:601 arg2 domain 620:625 only the sPLA(2) domain 603:625 only the sPLA(2) domain 603:625 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 1 78 theme group 136:140 arg1 2 171:171 2 171:171 2 171:171 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 78 theme group 136:140 arg1 sPLA 175:178 sPLA(2)-III 175:185 sPLA(2)-III 175:185 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 1 78 theme group 136:140 arg1 A 169:169 Human group III secreted phospholipase A 130:169 Human group III secreted phospholipase A(2) (sPLA(2)-III) 130:186 Human group III secreted phospholipase A(2) (sPLA(2)-III) consists of a central group III sPLA(2) domain flanked by unique N- and C-terminal domains. 15863501 3 79 theme form 587:590 arg1 production 569:578 the production 565:578 the production 565:578 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15863501 3 79 theme form 587:590 arg1 N-glycosylated 651:664 N-glycosylated 651:664 N-glycosylated 651:664 In several if not all cell types, the N- and C-terminal domains of sPLA(2)-III were proteolytically removed, leading to the production of the form containing only the sPLA(2) domain, which could be further N-glycosylated at two consensus sites. 15458386 7 0 theme C-terminal 970:979 arg1 enzyme 998:1003 the C-terminal domain-truncated enzyme 966:1003 the C-terminal domain-truncated enzyme 966:1003 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 8 1 theme endosome-like 1105:1117 arg1 structures 1119:1128 endosome-like structures 1105:1128 endosome-like structures 1105:1128 Deglycosylation blocked the release of the enzyme that accumulated in endosome-like structures. 15458386 9 2 theme enzyme 1135:1140 arg1 activity 1142:1149 The enzyme activity 1131:1149 The enzyme activity 1131:1149 The enzyme activity was also decreased by mutations of the residues forming the putative metal-binding sites and the active core. 15458386 3 3 theme amino 340:344 arg1 residues 351:358 The amino acid residues 336:358 The amino acid residues forming the two metal-binding sites in NPP 336:401 The amino acid residues forming the two metal-binding sites in NPP are conserved, and those of the active core are modified. 15458386 11 4 gly N-glycosylation 1525:1539 arg1 core 1629:1632 the predicted active core 1608:1632 the predicted active core 1608:1632 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 11 4 gly N-glycosylation 1525:1539 arg1 sites 1598:1602 the putative metal-binding sites 1571:1602 the putative metal-binding sites 1571:1602 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 11 5 theme sites 1598:1602 arg1 N-glycosylation 1525:1539 defective N-glycosylation 1515:1539 defective N-glycosylation 1515:1539 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 11 5 theme sites 1598:1602 arg1 alterations 1556:1566 structural alterations 1545:1566 structural alterations of the putative metal-binding sites and the predicted active core 1545:1632 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 10 6 theme sequence 1293:1300 arg1 Substitution 1261:1272 Substitution 1261:1272 Substitution of the active core sequence with that of NPP or mutation of T75 in the core 1261:1348 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 4 7 theme enzyme 503:508 arg1 changes 488:494 the functional changes 473:494 the functional changes of the enzyme induced by deglycosylation and mutagenesis 473:551 We examined the functional changes of the enzyme induced by deglycosylation and mutagenesis. 15458386 11 8 theme active 1622:1627 arg1 core 1629:1632 the predicted active core 1608:1632 the predicted active core 1608:1632 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 10 9 with Substitution 1261:1272 arg1 that 1307:1310 that 1307:1310 that 1307:1310 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 7 10 theme plasma 941:946 arg1 membrane 948:955 the plasma membrane 937:955 the plasma membrane 937:955 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 6 11 gly glycosylated 789:800 arg1 sites 774:778 these sites 768:778 these sites 768:778 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 3 12 theme metal-binding 376:388 arg1 sites 390:394 the two metal-binding sites 368:394 the two metal-binding sites in NPP 368:401 The amino acid residues forming the two metal-binding sites in NPP are conserved, and those of the active core are modified. 15458386 3 13 from sites 390:394 arg1 NPP 399:401 NPP 399:401 NPP 399:401 The amino acid residues forming the two metal-binding sites in NPP are conserved, and those of the active core are modified. 15458386 9 14 theme putative 1211:1218 arg1 sites 1234:1238 the putative metal-binding sites 1207:1238 the putative metal-binding sites 1207:1238 The enzyme activity was also decreased by mutations of the residues forming the putative metal-binding sites and the active core. 15458386 5 15 theme expressed 632:640 arg1 enzyme 642:647 the expressed enzyme 628:647 the expressed enzyme 628:647 Treating alk-SMase cDNA-transfected COS-7 cells with tunicamycin rendered the expressed enzyme completely inactive. 15458386 9 16 theme residues 1190:1197 arg1 mutations 1173:1181 mutations 1173:1181 mutations of the residues forming the putative metal-binding sites and the active core 1173:1258 The enzyme activity was also decreased by mutations of the residues forming the putative metal-binding sites and the active core. 15458386 10 17 theme active 1281:1286 arg1 sequence 1293:1300 the active core sequence 1277:1300 the active core sequence 1277:1300 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 10 18 theme T75 1334:1336 arg1 NPP 1315:1317 NPP 1315:1317 NPP 1315:1317 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 10 18 theme T75 1334:1336 arg1 mutation 1322:1329 mutation 1322:1329 mutation 1322:1329 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 7 19 from membrane 948:955 arg1 enzyme 908:913 the wild-type enzyme 894:913 the wild-type enzyme 894:913 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 7 19 from membrane 948:955 arg1 located 926:932 located 926:932 located 926:932 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 11 20 theme defective 1515:1523 arg1 N-glycosylation 1525:1539 defective N-glycosylation 1515:1539 defective N-glycosylation 1515:1539 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 1 21 theme NPP 182:184 arg1 family 217:222 the NPP (nucleotide phosphodiesterase) family 178:222 the NPP (nucleotide phosphodiesterase) family 178:222 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 11 22 theme alk-SMase 1472:1480 arg1 activity 1482:1489 alk-SMase activity 1472:1489 alk-SMase activity 1472:1489 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 5 23 theme COS-7 590:594 arg1 cells 596:600 alk-SMase cDNA-transfected COS-7 cells 563:600 alk-SMase cDNA-transfected COS-7 cells 563:600 Treating alk-SMase cDNA-transfected COS-7 cells with tunicamycin rendered the expressed enzyme completely inactive. 15458386 10 24 from NPP 1315:1317 arg1 core 1345:1348 the core 1341:1348 the core 1341:1348 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 4 25 theme functional 477:486 arg1 changes 488:494 the functional changes 473:494 the functional changes of the enzyme induced by deglycosylation and mutagenesis 473:551 We examined the functional changes of the enzyme induced by deglycosylation and mutagenesis. 15458386 6 26 theme enzyme 844:849 arg1 activity 851:858 the enzyme activity 840:858 the enzyme activity 840:858 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 1 27 theme Intestinal 101:110 arg1 alk-SMase 112:120 Intestinal alk-SMase 101:120 Intestinal alk-SMase (alkaline sphingomyelinase) 101:148 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 1 27 theme Intestinal 101:110 arg1 sphingomyelinase 132:147 alkaline sphingomyelinase 123:147 alkaline sphingomyelinase 123:147 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 1 27 theme Intestinal 101:110 arg1 ectoenzyme 156:165 an ectoenzyme 153:165 an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family 153:222 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 1 28 theme nucleotide 187:196 arg1 family 217:222 the NPP (nucleotide phosphodiesterase) family 178:222 the NPP (nucleotide phosphodiesterase) family 178:222 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 0 29 theme Functional 0:9 arg1 studies 11:17 Functional studies 0:17 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis 0:98 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis. 15458386 10 30 from mutation 1322:1329 arg1 core 1345:1348 the core 1341:1348 the core 1341:1348 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 7 31 theme wild-type 898:906 arg1 enzyme 908:913 the wild-type enzyme 894:913 the wild-type enzyme 894:913 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 7 31 theme wild-type 898:906 arg1 located 926:932 located 926:932 located 926:932 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 1 32 theme phosphodiesterase 198:214 arg1 family 217:222 the NPP (nucleotide phosphodiesterase) family 178:222 the NPP (nucleotide phosphodiesterase) family 178:222 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 0 33 theme human 22:26 arg1 sphingomyelinase 48:63 human intestinal alkaline sphingomyelinase 22:63 human intestinal alkaline sphingomyelinase 22:63 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis. 15458386 11 34 theme core 1629:1632 arg1 N-glycosylation 1525:1539 defective N-glycosylation 1515:1539 defective N-glycosylation 1515:1539 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 11 34 theme core 1629:1632 arg1 alterations 1556:1566 structural alterations 1545:1566 structural alterations of the putative metal-binding sites and the predicted active core 1545:1632 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 11 35 theme putative 1575:1582 arg1 sites 1598:1602 the putative metal-binding sites 1571:1602 the putative metal-binding sites 1571:1602 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 11 36 theme predicted 1612:1620 arg1 core 1629:1632 the predicted active core 1608:1632 the predicted active core 1608:1632 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 9 37 theme metal-binding 1220:1232 arg1 sites 1234:1238 the putative metal-binding sites 1207:1238 the putative metal-binding sites 1207:1238 The enzyme activity was also decreased by mutations of the residues forming the putative metal-binding sites and the active core. 15458386 1 38 theme alkaline 123:130 arg1 alk-SMase 112:120 Intestinal alk-SMase 101:120 Intestinal alk-SMase (alkaline sphingomyelinase) 101:148 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 1 38 theme alkaline 123:130 arg1 sphingomyelinase 132:147 alkaline sphingomyelinase 123:147 alkaline sphingomyelinase 123:147 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 6 39 theme sites 718:722 arg1 Mutations 670:678 Mutations 670:678 Mutations of the five potential N-glycosylation sites individually and in combination 670:754 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 11 40 theme structural 1545:1554 arg1 alterations 1556:1566 structural alterations 1545:1566 structural alterations of the putative metal-binding sites and the predicted active core 1545:1632 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 0 41 theme alkaline 39:46 arg1 sphingomyelinase 48:63 human intestinal alkaline sphingomyelinase 22:63 human intestinal alkaline sphingomyelinase 22:63 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis. 15458386 7 42 theme Immunogold 861:870 arg1 labelling 872:880 Immunogold labelling 861:880 Immunogold labelling 861:880 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 2 43 theme N-glycosylation 247:261 arg1 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 6 44 theme N-glycosylation 702:716 arg1 sites 718:722 the five potential N-glycosylation sites 683:722 the five potential N-glycosylation sites individually and in combination 683:754 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 0 45 theme intestinal 28:37 arg1 sphingomyelinase 48:63 human intestinal alkaline sphingomyelinase 22:63 human intestinal alkaline sphingomyelinase 22:63 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis. 15458386 2 46 gly N-glycosylation 247:261 arg2 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 2 46 gly N-glycosylation 247:261 arg2 five 232:235 five 232:235 five 232:235 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 6 47 gly N-glycosylation 702:716 arg2 five 687:690 five 687:690 five 687:690 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 6 47 gly N-glycosylation 702:716 arg2 sites 718:722 the five potential N-glycosylation sites 683:722 the five potential N-glycosylation sites individually and in combination 683:754 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 2 48 theme potential 237:245 arg1 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 10 49 theme enzyme 1364:1369 arg1 activity 1371:1378 the enzyme activity 1360:1378 the enzyme activity against sphingomyelin 1360:1400 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 3 50 theme active 435:440 arg1 core 442:445 the active core 431:445 the active core 431:445 The amino acid residues forming the two metal-binding sites in NPP are conserved, and those of the active core are modified. 15458386 6 51 theme deficient 806:814 arg1 glycosylation 816:828 deficient glycosylation 806:828 deficient glycosylation 806:828 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 0 52 theme sphingomyelinase 48:63 arg1 studies 11:17 Functional studies 0:17 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis 0:98 Functional studies of human intestinal alkaline sphingomyelinase by deglycosylation and mutagenesis. 15458386 7 53 located located 926:932 arg1 membrane 948:955 the plasma membrane 937:955 the plasma membrane 937:955 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 7 53 located located 926:932 arg2 located 926:932 located 926:932 located 926:932 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 7 53 located located 926:932 arg2 enzyme 908:913 the wild-type enzyme 894:913 the wild-type enzyme 894:913 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 5 54 theme alk-SMase 563:571 arg1 cells 596:600 alk-SMase cDNA-transfected COS-7 cells 563:600 alk-SMase cDNA-transfected COS-7 cells 563:600 Treating alk-SMase cDNA-transfected COS-7 cells with tunicamycin rendered the expressed enzyme completely inactive. 15458386 2 55 dep N- 318:319 arg1 the 314:316 the 314:316 the 314:316 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 6 56 theme potential 692:700 arg1 sites 718:722 the five potential N-glycosylation sites 683:722 the five potential N-glycosylation sites individually and in combination 683:754 Mutations of the five potential N-glycosylation sites individually and in combination showed that these sites were all glycosylated and deficient glycosylation decreased the enzyme activity. 15458386 8 57 theme enzyme 1078:1083 arg1 release 1063:1069 the release 1059:1069 the release of the enzyme that accumulated in endosome-like structures 1059:1128 Deglycosylation blocked the release of the enzyme that accumulated in endosome-like structures. 15458386 2 58 theme transmembrane 284:296 arg1 domains 298:304 predicated transmembrane domains 273:304 predicated transmembrane domains 273:304 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 5 59 theme cDNA-transfected 573:588 arg1 cells 596:600 alk-SMase cDNA-transfected COS-7 cells 563:600 alk-SMase cDNA-transfected COS-7 cells 563:600 Treating alk-SMase cDNA-transfected COS-7 cells with tunicamycin rendered the expressed enzyme completely inactive. 15458386 7 60 theme domain-truncated 981:996 arg1 enzyme 998:1003 the C-terminal domain-truncated enzyme 966:1003 the C-terminal domain-truncated enzyme 966:1003 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 2 61 contain has 228:230 arg2 domains 298:304 predicated transmembrane domains 273:304 predicated transmembrane domains 273:304 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 2 61 contain has 228:230 arg2 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 2 61 contain has 228:230 arg1 It 225:226 It 225:226 It 225:226 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 2 62 theme predicated 273:282 arg1 domains 298:304 predicated transmembrane domains 273:304 predicated transmembrane domains 273:304 It has five potential N-glycosylation sites and predicated transmembrane domains at both the N- and C-termini. 15458386 10 63 theme active 1438:1443 arg1 enzyme 1427:1432 the enzyme 1423:1432 the enzyme NPP active 1423:1443 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 3 64 theme acid 346:349 arg1 residues 351:358 The amino acid residues 336:358 The amino acid residues forming the two metal-binding sites in NPP 336:401 The amino acid residues forming the two metal-binding sites in NPP are conserved, and those of the active core are modified. 15458386 7 65 from located 926:932 arg1 membrane 948:955 the plasma membrane 937:955 the plasma membrane 937:955 Immunogold labelling showed that the wild-type enzyme was mainly located in the plasma membrane, whereas the C-terminal domain-truncated enzyme was released into the medium. 15458386 11 66 theme metal-binding 1584:1596 arg1 sites 1598:1602 the putative metal-binding sites 1571:1602 the putative metal-binding sites 1571:1602 Our results indicate that alk-SMase activity is severely affected by defective N-glycosylation and structural alterations of the putative metal-binding sites and the predicted active core. 15458386 10 67 theme core 1288:1291 arg1 sequence 1293:1300 the active core sequence 1277:1300 the active core sequence 1277:1300 Substitution of the active core sequence with that of NPP or mutation of T75 in the core abolished the enzyme activity against sphingomyelin but failed to render the enzyme NPP active. 15458386 9 68 theme active 1248:1253 arg1 core 1255:1258 the active core 1244:1258 the active core 1244:1258 The enzyme activity was also decreased by mutations of the residues forming the putative metal-binding sites and the active core. 15458386 1 69 theme related 167:173 arg1 alk-SMase 112:120 Intestinal alk-SMase 101:120 Intestinal alk-SMase (alkaline sphingomyelinase) 101:148 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 15458386 1 69 theme related 167:173 arg1 ectoenzyme 156:165 an ectoenzyme 153:165 an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family 153:222 Intestinal alk-SMase (alkaline sphingomyelinase) is an ectoenzyme related to the NPP (nucleotide phosphodiesterase) family. 8702538 10 0 from position 1684:1691 arg1 located 1663:1669 located 1663:1669 located 1663:1669 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 0 from position 1684:1691 arg1 glycoproteins 1714:1726 the CD22 and CD33 glycoproteins 1696:1726 the CD22 and CD33 glycoproteins 1696:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 1 1 theme I-type 246:251 arg1 class 253:257 the recently identified I-type class 222:257 the recently identified I-type class of lectins 222:268 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 0 2 link N-linked 9:16 arg1 site 32:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 2 3 theme Recent 271:276 arg1 work 278:281 Recent work 271:281 Recent work 271:281 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 8 4 theme second 1314:1319 arg1 domains 1324:1330 the first or second Ig domains 1301:1330 the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1301:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 4 theme second 1314:1319 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 7 5 theme sequence 1154:1161 arg1 NCT 1163:1165 the sequence NCT 1150:1165 the sequence NCT 1150:1165 Interestingly, this site is characterized by the sequence NCT, where the cysteine is thought to be involved in an intrachain disulfide bond. 8702538 1 6 theme superfamily 154:164 arg1 receptor 197:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 6 theme superfamily 154:164 arg1 CD22 128:131 CD22 128:131 CD22 128:131 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 10 7 link N-linked 1635:1642 arg1 critical 1731:1738 critical 1731:1738 critical 1731:1738 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 7 link N-linked 1635:1642 arg1 site 1658:1661 a single N-linked glycosylation site 1626:1661 a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins 1626:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 1 8 theme lymphocyte-specific 168:186 arg1 receptor 197:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 8 theme lymphocyte-specific 168:186 arg1 CD22 128:131 CD22 128:131 CD22 128:131 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 6 9 theme site 1016:1019 arg1 mutation 962:969 mutation 962:969 mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 962:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 10 10 theme similar 1676:1682 arg1 position 1684:1691 a similar position 1674:1691 a similar position in the CD22 and CD33 glycoproteins 1674:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 3 11 theme CD22 597:600 arg1 receptor 602:609 the CD22 receptor 593:609 the CD22 receptor itself 593:616 CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. 8702538 6 12 theme N-linked 993:1000 arg1 site 1016:1019 a single potential N-linked glycosylation site 974:1019 a single potential N-linked glycosylation site 974:1019 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 10 13 theme glycosylation 1644:1656 arg1 critical 1731:1738 critical 1731:1738 critical 1731:1738 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 13 theme glycosylation 1644:1656 arg1 site 1658:1661 a single N-linked glycosylation site 1626:1661 a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins 1626:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 8 14 theme first 1305:1309 arg1 domains 1324:1330 the first or second Ig domains 1301:1330 the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1301:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 14 theme first 1305:1309 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 10 15 theme single 1628:1633 arg1 critical 1731:1738 critical 1731:1738 critical 1731:1738 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 15 theme single 1628:1633 arg1 site 1658:1661 a single N-linked glycosylation site 1626:1661 a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins 1626:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 6 16 link N-linked 993:1000 arg1 site 1016:1019 a single potential N-linked glycosylation site 974:1019 a single potential N-linked glycosylation site 974:1019 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 9 17 theme motif 1496:1500 arg1 mutation 1478:1485 mutation 1478:1485 mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 1478:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 8 18 theme similar 1275:1281 arg1 motifs 1291:1296 similar NC(T/S) motifs 1275:1296 similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1275:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 6 19 theme single 976:981 arg1 site 1016:1019 a single potential N-linked glycosylation site 974:1019 a single potential N-linked glycosylation site 974:1019 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 4 20 theme adhesion 752:759 arg1 regulation 724:733 the regulation 720:733 the regulation of CD22-mediated adhesion 720:759 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 5 21 from mutagenesis 807:817 arg1 CD22 864:867 CD22 864:867 CD22 864:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 5 22 link N-linked 832:839 arg1 sites 855:859 potential N-linked glycosylation sites 822:859 potential N-linked glycosylation sites on CD22 822:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 10 23 gly glycosylation 1644:1656 arg2 critical 1731:1738 critical 1731:1738 critical 1731:1738 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 23 gly glycosylation 1644:1656 arg2 site 1658:1661 a single N-linked glycosylation site 1626:1661 a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins 1626:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 9 24 theme Ig 1515:1516 arg1 CD33 1546:1549 the I-type lectin CD33 1528:1549 the I-type lectin CD33 1528:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 9 24 theme Ig 1515:1516 arg1 domain 1518:1523 the first Ig domain 1505:1523 the first Ig domain of the I-type lectin CD33 1505:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 5 25 theme site-specific 793:805 arg1 mutagenesis 807:817 site-specific mutagenesis 793:817 site-specific mutagenesis of potential N-linked glycosylation sites on CD22 793:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 5 26 theme ligand 929:934 arg1 recognition 936:946 ligand recognition 929:946 ligand recognition 929:946 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 2 27 gly glycoproteins 418:430 arg1 glycoproteins 418:430 selected cell surface glycoproteins 396:430 selected cell surface glycoproteins 396:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 8 28 theme lectins 1346:1352 arg1 domains 1324:1330 the first or second Ig domains 1301:1330 the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1301:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 28 theme lectins 1346:1352 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 10 29 gly glycoproteins 1714:1726 arg1 glycoproteins 1714:1726 the CD22 and CD33 glycoproteins 1696:1726 the CD22 and CD33 glycoproteins 1696:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 8 30 from domains 1324:1330 arg1 mutagenesis 1260:1270 Site-directed mutagenesis 1246:1270 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1246:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 0 31 dep lectins 105:111 arg1 lectins 105:111 the I-type lectins CD22 and CD33 94:125 the I-type lectins CD22 and CD33 94:125 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 0 31 dep lectins 105:111 arg1 CD33 122:125 CD33 122:125 CD33 122:125 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 0 31 dep lectins 105:111 arg1 CD22 113:116 CD22 113:116 CD22 113:116 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 6 32 theme immunoglobulin 1034:1047 arg1 CD22 1059:1062 CD22 1059:1062 CD22 1059:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 6 32 theme immunoglobulin 1034:1047 arg1 domain 1049:1054 the first immunoglobulin domain 1024:1054 the first immunoglobulin domain of CD22 1024:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 0 33 theme recognition 79:89 arg1 regulation 58:67 the regulation 54:67 the regulation of ligand recognition by the I-type lectins CD22 and CD33 54:125 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 4 34 gly glycosylation 660:672 arg2 sites 674:678 N-linked glycosylation sites 651:678 N-linked glycosylation sites on the CD22 molecule 651:699 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 9 35 theme CD33 1546:1549 arg1 CD33 1546:1549 the I-type lectin CD33 1528:1549 the I-type lectin CD33 1528:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 9 35 theme CD33 1546:1549 arg1 domain 1518:1523 the first Ig domain 1505:1523 the first Ig domain of the I-type lectin CD33 1505:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 2 36 theme selected 396:403 arg1 glycoproteins 418:430 selected cell surface glycoproteins 396:430 selected cell surface glycoproteins 396:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 7 37 theme disulfide 1230:1238 arg1 bond 1240:1243 an intrachain disulfide bond 1216:1243 an intrachain disulfide bond 1216:1243 Interestingly, this site is characterized by the sequence NCT, where the cysteine is thought to be involved in an intrachain disulfide bond. 8702538 3 38 theme CD22-ligand 433:443 arg1 interaction 445:455 CD22-ligand interaction 433:455 CD22-ligand interaction 433:455 CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. 8702538 1 39 theme lectins 262:268 arg1 class 253:257 the recently identified I-type class 222:257 the recently identified I-type class of lectins 222:268 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 9 40 theme acid 1571:1574 arg1 activity 1584:1591 its sialic acid binding activity 1560:1591 its sialic acid binding activity 1560:1591 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 0 41 theme N-linked 9:16 arg1 site 32:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 10 42 theme ligand 1755:1760 arg1 recognition 1762:1772 ligand recognition 1755:1772 ligand recognition by both receptors 1755:1790 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 2 43 link N-linked 367:374 arg1 oligosaccharides 376:391 terminal N-linked oligosaccharides 358:391 terminal N-linked oligosaccharides on selected cell surface glycoproteins 358:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 2 44 theme surface 410:416 arg1 glycoproteins 418:430 selected cell surface glycoproteins 396:430 selected cell surface glycoproteins 396:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 5 45 theme N-linked 832:839 arg1 sites 855:859 potential N-linked glycosylation sites 822:859 potential N-linked glycosylation sites on CD22 822:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 4 46 link N-linked 651:658 arg1 sites 674:678 N-linked glycosylation sites 651:678 N-linked glycosylation sites on the CD22 molecule 651:699 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 2 47 from oligosaccharides 376:391 arg1 glycoproteins 418:430 selected cell surface glycoproteins 396:430 selected cell surface glycoproteins 396:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 6 48 theme ligand 1085:1090 arg1 recognition 1092:1102 ligand recognition 1085:1102 ligand recognition 1085:1102 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 3 49 theme alpha2 508:513 arg1 activity 477:484 the activity 473:484 the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself 473:616 CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. 8702538 5 50 theme sites 855:859 arg1 mutagenesis 807:817 site-specific mutagenesis 793:817 site-specific mutagenesis of potential N-linked glycosylation sites on CD22 793:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 0 51 gly glycosylation 18:30 arg2 site 32:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 8 52 theme acid 1451:1454 arg1 binding 1456:1462 sialic acid binding 1444:1462 sialic acid binding 1444:1462 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 10 53 theme CD22 1700:1703 arg1 glycoproteins 1714:1726 the CD22 and CD33 glycoproteins 1696:1726 the CD22 and CD33 glycoproteins 1696:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 2 54 theme terminal 358:365 arg1 oligosaccharides 376:391 terminal N-linked oligosaccharides 358:391 terminal N-linked oligosaccharides on selected cell surface glycoproteins 358:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 4 55 theme glycosylation 660:672 arg1 sites 674:678 N-linked glycosylation sites 651:678 N-linked glycosylation sites on the CD22 molecule 651:699 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 6 56 from mutation 962:969 arg1 CD22 1059:1062 CD22 1059:1062 CD22 1059:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 6 56 from mutation 962:969 arg1 domain 1049:1054 the first immunoglobulin domain 1024:1054 the first immunoglobulin domain of CD22 1024:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 1 57 theme identified 235:244 arg1 class 253:257 the recently identified I-type class 222:257 the recently identified I-type class of lectins 222:268 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 2 58 attach linked 339:344 arg2 acid 334:337 sialic acid 327:337 sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins 327:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 2 58 attach linked 339:344 arg1 oligosaccharides 376:391 terminal N-linked oligosaccharides 358:391 terminal N-linked oligosaccharides on selected cell surface glycoproteins 358:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 1 59 theme class 253:257 arg1 receptor 197:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 59 theme class 253:257 arg1 member 212:217 a member 210:217 a member of the recently identified I-type class of lectins 210:268 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 59 theme class 253:257 arg1 CD22 128:131 CD22 128:131 CD22 128:131 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 10 60 theme CD33 1709:1712 arg1 glycoproteins 1714:1726 the CD22 and CD33 glycoproteins 1696:1726 the CD22 and CD33 glycoproteins 1696:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 3 61 theme CD22-mediated 556:568 arg1 binding 570:576 CD22-mediated binding 556:576 CD22-mediated binding 556:576 CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. 8702538 1 62 theme immunoglobulin 139:152 arg1 receptor 197:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 62 theme immunoglobulin 139:152 arg1 CD22 128:131 CD22 128:131 CD22 128:131 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 8 63 from mutagenesis 1260:1270 arg1 domains 1324:1330 the first or second Ig domains 1301:1330 the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1301:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 63 from mutagenesis 1260:1270 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 64 theme NC 1283:1284 arg1 motifs 1291:1296 similar NC(T/S) motifs 1275:1296 similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1275:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 1 65 theme B 166:166 arg1 receptor 197:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 65 theme B 166:166 arg1 CD22 128:131 CD22 128:131 CD22 128:131 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 8 66 theme T/S 1286:1288 arg1 motifs 1291:1296 similar NC(T/S) motifs 1275:1296 similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1275:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 6 67 gly glycosylation 1002:1014 arg2 site 1016:1019 a single potential N-linked glycosylation site 974:1019 a single potential N-linked glycosylation site 974:1019 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 1 68 theme adhesion 188:195 arg1 receptor 197:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 an immunoglobulin superfamily B lymphocyte-specific adhesion receptor 136:204 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 1 68 theme adhesion 188:195 arg1 CD22 128:131 CD22 128:131 CD22 128:131 CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. 8702538 6 69 theme glycosylation 1002:1014 arg1 site 1016:1019 a single potential N-linked glycosylation site 974:1019 a single potential N-linked glycosylation site 974:1019 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 9 70 theme I-type 1532:1537 arg1 CD33 1546:1549 the I-type lectin CD33 1528:1549 the I-type lectin CD33 1528:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 8 71 theme motifs 1291:1296 arg1 mutagenesis 1260:1270 Site-directed mutagenesis 1246:1270 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1246:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 10 72 theme N-linked 1635:1642 arg1 critical 1731:1738 critical 1731:1738 critical 1731:1738 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 72 theme N-linked 1635:1642 arg1 site 1658:1661 a single N-linked glycosylation site 1626:1661 a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins 1626:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 9 73 theme NCS 1492:1494 arg1 motif 1496:1500 a NCS motif 1490:1500 a NCS motif 1490:1500 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 8 74 theme Site-directed 1246:1258 arg1 mutagenesis 1260:1270 Site-directed mutagenesis 1246:1270 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1246:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 6 75 theme potential 983:991 arg1 site 1016:1019 a single potential N-linked glycosylation site 974:1019 a single potential N-linked glycosylation site 974:1019 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 4 76 theme CD22 687:690 arg1 molecule 692:699 the CD22 molecule 683:699 the CD22 molecule 683:699 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 4 77 theme CD22-mediated 738:750 arg1 adhesion 752:759 CD22-mediated adhesion 738:759 CD22-mediated adhesion 738:759 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 9 78 theme first 1509:1513 arg1 CD33 1546:1549 the I-type lectin CD33 1528:1549 the I-type lectin CD33 1528:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 9 78 theme first 1509:1513 arg1 domain 1518:1523 the first Ig domain 1505:1523 the first Ig domain of the I-type lectin CD33 1505:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 8 79 theme Ig 1321:1322 arg1 domains 1324:1330 the first or second Ig domains 1301:1330 the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1301:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 79 theme Ig 1321:1322 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 4 80 from sites 674:678 arg1 molecule 692:699 the CD22 molecule 683:699 the CD22 molecule 683:699 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 6 81 theme CD22 1059:1062 arg1 CD22 1059:1062 CD22 1059:1062 CD22 1059:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 6 81 theme CD22 1059:1062 arg1 domain 1049:1054 the first immunoglobulin domain 1024:1054 the first immunoglobulin domain of CD22 1024:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 0 82 theme ligand 72:77 arg1 recognition 79:89 ligand recognition 72:89 ligand recognition 72:89 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 5 83 from sites 855:859 arg1 CD22 864:867 CD22 864:867 CD22 864:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 8 84 from motifs 1291:1296 arg1 domains 1324:1330 the first or second Ig domains 1301:1330 the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1301:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 84 from motifs 1291:1296 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 85 theme I-type 1339:1344 arg1 glycoprotein 1372:1383 myelin-associated glycoprotein 1354:1383 myelin-associated glycoprotein 1354:1383 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 85 theme I-type 1339:1344 arg1 sialoadhesin 1390:1401 sialoadhesin 1390:1401 sialoadhesin 1390:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 85 theme I-type 1339:1344 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 9 86 theme lectin 1539:1544 arg1 CD33 1546:1549 the I-type lectin CD33 1528:1549 the I-type lectin CD33 1528:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 6 87 theme first 1028:1032 arg1 CD22 1059:1062 CD22 1059:1062 CD22 1059:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 6 87 theme first 1028:1032 arg1 domain 1049:1054 the first immunoglobulin domain 1024:1054 the first immunoglobulin domain of CD22 1024:1062 We show that mutation of a single potential N-linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. 8702538 8 88 theme myelin-associated 1354:1370 arg1 glycoprotein 1372:1383 myelin-associated glycoprotein 1354:1383 myelin-associated glycoprotein 1354:1383 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 88 theme myelin-associated 1354:1370 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 0 89 theme I-type 98:103 arg1 lectins 105:111 the I-type lectins CD22 and CD33 94:125 the I-type lectins CD22 and CD33 94:125 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 0 89 theme I-type 98:103 arg1 CD33 122:125 CD33 122:125 CD33 122:125 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 0 89 theme I-type 98:103 arg1 CD22 113:116 CD22 113:116 CD22 113:116 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 7 90 theme intrachain 1219:1228 arg1 bond 1240:1243 an intrachain disulfide bond 1216:1243 an intrachain disulfide bond 1216:1243 Interestingly, this site is characterized by the sequence NCT, where the cysteine is thought to be involved in an intrachain disulfide bond. 8702538 0 91 theme single 2:7 arg1 site 32:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 5 92 from CD22 864:867 arg1 mutagenesis 807:817 site-specific mutagenesis 793:817 site-specific mutagenesis of potential N-linked glycosylation sites on CD22 793:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 9 93 theme sialic 1564:1569 arg1 activity 1584:1591 its sialic acid binding activity 1560:1591 its sialic acid binding activity 1560:1591 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 2 94 theme N-linked 367:374 arg1 oligosaccharides 376:391 terminal N-linked oligosaccharides 358:391 terminal N-linked oligosaccharides on selected cell surface glycoproteins 358:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 0 95 theme glycosylation 18:30 arg1 site 32:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site 0:35 A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. 8702538 8 96 gly glycoprotein 1372:1383 arg1 glycoprotein 1372:1383 myelin-associated glycoprotein 1354:1383 myelin-associated glycoprotein 1354:1383 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 96 gly glycoprotein 1372:1383 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 9 97 theme binding 1576:1582 arg1 activity 1584:1591 its sialic acid binding activity 1560:1591 its sialic acid binding activity 1560:1591 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 5 98 theme potential 822:830 arg1 sites 855:859 potential N-linked glycosylation sites 822:859 potential N-linked glycosylation sites on CD22 822:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 8 99 dep lectins 1346:1352 arg1 glycoprotein 1372:1383 myelin-associated glycoprotein 1354:1383 myelin-associated glycoprotein 1354:1383 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 99 dep lectins 1346:1352 arg1 sialoadhesin 1390:1401 sialoadhesin 1390:1401 sialoadhesin 1390:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 8 99 dep lectins 1346:1352 arg1 lectins 1346:1352 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 the I-type lectins myelin-associated glycoprotein, and sialoadhesin 1335:1401 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 2 100 theme cell 405:408 arg1 glycoproteins 418:430 selected cell surface glycoproteins 396:430 selected cell surface glycoproteins 396:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 5 101 theme glycosylation 841:853 arg1 sites 855:859 potential N-linked glycosylation sites 822:859 potential N-linked glycosylation sites on CD22 822:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 8702538 2 102 theme sialic 327:332 arg1 acid 334:337 sialic acid 327:337 sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins 327:430 Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N-linked oligosaccharides on selected cell surface glycoproteins. 8702538 4 103 theme N-linked 651:658 arg1 sites 674:678 N-linked glycosylation sites 651:678 N-linked glycosylation sites on the CD22 molecule 651:699 These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. 8702538 8 104 theme sialic 1444:1449 arg1 binding 1456:1462 sialic acid binding 1444:1462 sialic acid binding 1444:1462 Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. 8702538 3 105 theme beta-galactoside 491:506 arg1 alpha2 508:513 a beta-galactoside alpha2 489:513 a beta-galactoside alpha2 489:513 CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. 8702538 3 105 theme beta-galactoside 491:506 arg1 6-sialyltransferase 516:534 6-sialyltransferase 516:534 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself 516:616 CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6-sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. 8702538 9 106 from mutation 1478:1485 arg1 CD33 1546:1549 the I-type lectin CD33 1528:1549 the I-type lectin CD33 1528:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 9 106 from mutation 1478:1485 arg1 domain 1518:1523 the first Ig domain 1505:1523 the first Ig domain of the I-type lectin CD33 1505:1549 In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. 8702538 10 107 theme located 1663:1669 arg1 critical 1731:1738 critical 1731:1738 critical 1731:1738 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 10 107 theme located 1663:1669 arg1 site 1658:1661 a single N-linked glycosylation site 1626:1661 a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins 1626:1726 These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. 8702538 5 108 gly glycosylation 841:853 arg2 sites 855:859 potential N-linked glycosylation sites 822:859 potential N-linked glycosylation sites on CD22 822:867 In this work we have performed site-specific mutagenesis of potential N-linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. 22040171 2 0 theme chromatographic 366:380 arg1 purification 382:393 a single step chromatographic purification 352:393 a single step chromatographic purification of the protein from the media 352:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 4 1 theme heterodimer 847:857 arg1 formation 827:835 formation 827:835 formation of an α/β heterodimer 827:857 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 2 2 theme step 361:364 arg1 purification 382:393 a single step chromatographic purification 352:393 a single step chromatographic purification of the protein from the media 352:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 6 3 theme cysteine 1061:1068 arg1 residues 1070:1077 the cysteine residues 1057:1077 the cysteine residues 1057:1077 The significance of the cysteine residues was established by their selective alkylation resulting in almost complete loss of activity. 22040171 4 4 theme enzyme 794:799 arg1 mass 775:778 the mass 771:778 the mass of the active enzyme 771:799 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 8 5 theme throughput 1368:1377 arg1 assay 1379:1383 a high throughput assay 1361:1383 a high throughput assay 1361:1383 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 8 5 theme throughput 1368:1377 arg1 useful 1394:1399 useful 1394:1399 useful 1394:1399 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 6 6 theme activity 1162:1169 arg1 loss 1154:1157 almost complete loss 1138:1157 almost complete loss of activity 1138:1169 The significance of the cysteine residues was established by their selective alkylation resulting in almost complete loss of activity. 22040171 8 7 theme quantities 1338:1347 arg1 production 1313:1322 The production 1309:1322 The production of sufficient quantities of NAAA 1309:1355 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 8 7 theme quantities 1338:1347 arg1 assay 1379:1383 a high throughput assay 1361:1383 a high throughput assay 1361:1383 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 8 7 theme quantities 1338:1347 arg1 useful 1394:1399 useful 1394:1399 useful 1394:1399 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 3 8 theme PNGase 589:594 arg1 F 586:586 peptide-N-glycosidase F 564:586 peptide-N-glycosidase F (PNGase F) 564:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 8 theme PNGase 589:594 arg1 F 596:596 PNGase F 589:596 PNGase F 589:596 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 1 9 theme N-Acylethanolamine-hydrolyzing 90:119 arg1 enzyme 156:161 a lysosomal enzyme 144:161 a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses 144:265 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 1 9 theme N-Acylethanolamine-hydrolyzing 90:119 arg1 NAAA 135:138 NAAA 135:138 NAAA 135:138 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 1 9 theme N-Acylethanolamine-hydrolyzing 90:119 arg1 amidase 126:132 N-Acylethanolamine-hydrolyzing acid amidase 90:132 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) 90:139 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 6 10 theme selective 1104:1112 arg1 alkylation 1114:1123 their selective alkylation 1098:1123 their selective alkylation resulting in almost complete loss of activity 1098:1169 The significance of the cysteine residues was established by their selective alkylation resulting in almost complete loss of activity. 22040171 1 11 theme acid 121:124 arg1 enzyme 156:161 a lysosomal enzyme 144:161 a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses 144:265 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 1 11 theme acid 121:124 arg1 NAAA 135:138 NAAA 135:138 NAAA 135:138 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 1 11 theme acid 121:124 arg1 amidase 126:132 N-Acylethanolamine-hydrolyzing acid amidase 90:132 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) 90:139 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 3 12 theme desorption/ionization 448:468 arg1 spectrometry 490:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 426:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 13 theme mass 485:488 arg1 spectrometry 490:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 426:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 14 theme Matrix-assisted 426:440 arg1 spectrometry 490:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 426:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 7 15 theme purified 1176:1183 arg1 enzyme 1185:1190 The purified enzyme 1172:1190 The purified enzyme 1172:1190 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 6 16 theme complete 1145:1152 arg1 loss 1154:1157 almost complete loss 1138:1157 almost complete loss of activity 1138:1169 The significance of the cysteine residues was established by their selective alkylation resulting in almost complete loss of activity. 22040171 3 17 theme spectrometry 490:501 arg1 analysis 516:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis 426:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 0 18 theme spectrometric 5:17 arg1 characterization 19:34 Mass spectrometric characterization 0:34 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 0:88 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 22040171 4 19 theme α/β 843:845 arg1 heterodimer 847:857 an α/β heterodimer 840:857 an α/β heterodimer 840:857 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 5 20 theme amino 917:921 arg1 sequence 928:935 the amino acid sequence 913:935 the amino acid sequence 913:935 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 8 21 theme enzyme 1484:1489 arg1 function 1467:1474 function 1467:1474 function 1467:1474 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 8 21 theme enzyme 1484:1489 arg1 structure 1453:1461 structure 1453:1461 structure 1453:1461 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 2 22 theme protein 402:408 arg1 pro-enzyme 327:336 the NAAA pro-enzyme 318:336 the NAAA pro-enzyme (zymogen) 318:346 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 2 22 theme protein 402:408 arg1 purification 382:393 a single step chromatographic purification 352:393 a single step chromatographic purification of the protein from the media 352:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 2 22 theme protein 402:408 arg1 zymogen 339:345 zymogen 339:345 zymogen 339:345 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 0 23 theme Mass 0:3 arg1 characterization 19:34 Mass spectrometric characterization 0:34 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 0:88 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 22040171 8 24 dep structure 1453:1461 arg1 the 1449:1451 the 1449:1451 the 1449:1451 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 4 25 theme exclusion 736:744 arg1 chromatography 746:759 Size exclusion chromatography 731:759 Size exclusion chromatography 731:759 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 3 26 theme time-of-flight 470:483 arg1 spectrometry 490:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 426:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 4 27 theme ± 807:807 arg1 kDa 811:813 45 ± 3 kDa 804:813 45 ± 3 kDa 804:813 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 8 28 theme novel 1416:1420 arg1 inhibitors 1422:1431 novel inhibitors 1416:1431 novel inhibitors 1416:1431 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 5 29 theme sequence 928:935 arg1 evidence 977:984 evidence 977:984 evidence for the lack of a disulfide bond between subunits 977:1034 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 5 29 theme sequence 928:935 arg1 % 908:908 more than 80% 896:908 more than 80% of the amino acid sequence, including the N-terminal peptides 896:970 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 5 29 theme sequence 928:935 arg1 sequence 928:935 the amino acid sequence 913:935 the amino acid sequence 913:935 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 1 30 theme lipid 218:222 arg1 amide 224:228 a lipid amide 216:228 a lipid amide that inhibits inflammatory responses 216:265 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 1 30 theme lipid 218:222 arg1 palmitoylethanolamine 187:207 palmitoylethanolamine 187:207 palmitoylethanolamine (PEA) 187:213 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 5 31 theme acid 923:926 arg1 sequence 928:935 the amino acid sequence 913:935 the amino acid sequence 913:935 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 3 32 theme peptide-N-glycosidase 564:584 arg1 F 586:586 peptide-N-glycosidase F 564:586 peptide-N-glycosidase F (PNGase F) 564:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 32 theme peptide-N-glycosidase 564:584 arg1 F 596:596 PNGase F 589:596 PNGase F 589:596 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 2 33 theme NAAA 322:325 arg1 pro-enzyme 327:336 the NAAA pro-enzyme 318:336 the NAAA pro-enzyme (zymogen) 318:346 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 2 33 theme NAAA 322:325 arg1 zymogen 339:345 zymogen 339:345 zymogen 339:345 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 0 34 theme N-acylethanolamine-hydrolyzing 45:74 arg1 amidase 81:87 human N-acylethanolamine-hydrolyzing acid amidase 39:87 human N-acylethanolamine-hydrolyzing acid amidase 39:87 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 22040171 7 35 theme 4-methyl 1269:1276 arg1 substrate 1255:1263 a novel fluorogenic substrate 1235:1263 a novel fluorogenic substrate 1235:1263 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 35 theme 4-methyl 1269:1276 arg1 PAMCA 1301:1305 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 5 36 theme MS 870:871 arg1 fingerprinting 873:886 MALDI-TOF MS fingerprinting 860:886 MALDI-TOF MS fingerprinting 860:886 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 3 37 theme MALDI-TOF 503:511 arg1 analysis 516:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis 426:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 0 38 theme human 39:43 arg1 amidase 81:87 human N-acylethanolamine-hydrolyzing acid amidase 39:87 human N-acylethanolamine-hydrolyzing acid amidase 39:87 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 22040171 3 39 theme laser 442:446 arg1 spectrometry 490:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 426:501 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 40 theme zymogen 658:664 arg1 cleavage 642:649 acid cleavage 637:649 acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) 637:707 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 41 gly glycosylation 612:624 arg2 4 610:610 4 610:610 4 610:610 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 41 gly glycosylation 612:624 arg2 sites 626:630 4 glycosylation sites 610:630 4 glycosylation sites 610:630 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 0 42 theme amidase 81:87 arg1 characterization 19:34 Mass spectrometric characterization 0:34 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 0:88 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 22040171 5 43 theme MALDI-TOF 860:868 arg1 fingerprinting 873:886 MALDI-TOF MS fingerprinting 860:886 MALDI-TOF MS fingerprinting 860:886 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 5 44 theme N-terminal 952:961 arg1 peptides 963:970 the N-terminal peptides 948:970 the N-terminal peptides 948:970 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 3 45 theme glycosylation 612:624 arg1 sites 626:630 4 glycosylation sites 610:630 4 glycosylation sites 610:630 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 8 46 theme sufficient 1327:1336 arg1 quantities 1338:1347 sufficient quantities 1327:1347 sufficient quantities of NAAA 1327:1355 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 2 47 theme single 354:359 arg1 step 361:364 a single step 352:364 a single step chromatographic purification of the protein from the media 352:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 0 48 theme acid 76:79 arg1 amidase 81:87 human N-acylethanolamine-hydrolyzing acid amidase 39:87 human N-acylethanolamine-hydrolyzing acid amidase 39:87 Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. 22040171 5 49 theme disulfide 1004:1012 arg1 bond 1014:1017 a disulfide bond 1002:1017 a disulfide bond between subunits 1002:1034 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 8 50 theme high 1363:1366 arg1 assay 1379:1383 a high throughput assay 1361:1383 a high throughput assay 1361:1383 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 8 50 theme high 1363:1366 arg1 useful 1394:1399 useful 1394:1399 useful 1394:1399 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 1 51 theme lysosomal 146:154 arg1 enzyme 156:161 a lysosomal enzyme 144:161 a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses 144:265 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 1 51 theme lysosomal 146:154 arg1 amidase 126:132 N-Acylethanolamine-hydrolyzing acid amidase 90:132 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) 90:139 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 7 52 theme novel 1237:1241 arg1 substrate 1255:1263 a novel fluorogenic substrate 1235:1263 a novel fluorogenic substrate 1235:1263 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 52 theme novel 1237:1241 arg1 PAMCA 1301:1305 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 5 53 theme bond 1014:1017 arg1 lack 994:997 the lack 990:997 the lack of a disulfide bond between subunits 990:1034 MALDI-TOF MS fingerprinting covered more than 80% of the amino acid sequence, including the N-terminal peptides, and evidence for the lack of a disulfide bond between subunits. 22040171 2 54 theme cell 290:293 arg1 line 295:298 a HEK293 cell line 281:298 a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media 281:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 4 55 theme Size 731:734 arg1 chromatography 746:759 Size exclusion chromatography 731:759 Size exclusion chromatography 731:759 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 8 56 theme NAAA 1352:1355 arg1 quantities 1338:1347 sufficient quantities 1327:1347 sufficient quantities of NAAA 1327:1355 The production of sufficient quantities of NAAA and a high throughput assay could be useful in discovering novel inhibitors and determining the structure and function of this enzyme. 22040171 1 57 theme inflammatory 244:255 arg1 responses 257:265 inflammatory responses 244:265 inflammatory responses 244:265 N-Acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal enzyme that primarily degrades palmitoylethanolamine (PEA), a lipid amide that inhibits inflammatory responses. 22040171 4 58 theme active 787:792 arg1 enzyme 794:799 the active enzyme 783:799 the active enzyme 783:799 Size exclusion chromatography estimated the mass of the active enzyme as 45 ± 3 kDa, suggesting formation of an α/β heterodimer. 22040171 2 59 theme HEK293 283:288 arg1 line 295:298 a HEK293 cell line 281:298 a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media 281:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 3 60 theme zymogen 532:538 arg1 analysis 516:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis 426:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 7 61 theme palmitamide 1288:1298 arg1 substrate 1255:1263 a novel fluorogenic substrate 1235:1263 a novel fluorogenic substrate 1235:1263 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 61 theme palmitamide 1288:1298 arg1 PAMCA 1301:1305 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 62 theme fluorogenic 1243:1253 arg1 substrate 1255:1263 a novel fluorogenic substrate 1235:1263 a novel fluorogenic substrate 1235:1263 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 62 theme fluorogenic 1243:1253 arg1 PAMCA 1301:1305 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 3 63 theme MS 513:514 arg1 analysis 516:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis 426:523 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) 426:597 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 3 64 theme acid 637:640 arg1 cleavage 642:649 acid cleavage 637:649 acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) 637:707 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MALDI-TOF MS analysis of the zymogen (47.7 kDa) treated with peptide-N-glycosidase F (PNGase F) identified 4 glycosylation sites, and acid cleavage of the zymogen into α- and β-subunits (14.6 and 33.3 kDa) activated the enzyme. 22040171 2 65 from media 419:423 arg1 pro-enzyme 327:336 the NAAA pro-enzyme 318:336 the NAAA pro-enzyme (zymogen) 318:346 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 2 65 from media 419:423 arg1 purification 382:393 a single step chromatographic purification 352:393 a single step chromatographic purification of the protein from the media 352:423 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 2 65 from media 419:423 arg1 zymogen 339:345 zymogen 339:345 zymogen 339:345 We developed a HEK293 cell line stably expressing the NAAA pro-enzyme (zymogen) and a single step chromatographic purification of the protein from the media. 22040171 7 66 theme coumarin 1278:1285 arg1 substrate 1255:1263 a novel fluorogenic substrate 1235:1263 a novel fluorogenic substrate 1235:1263 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 66 theme coumarin 1278:1285 arg1 PAMCA 1301:1305 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 67 theme N- 1266:1267 arg1 substrate 1255:1263 a novel fluorogenic substrate 1235:1263 a novel fluorogenic substrate 1235:1263 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 7 67 theme N- 1266:1267 arg1 PAMCA 1301:1305 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 N-(4-methyl coumarin) palmitamide (PAMCA) 1266:1306 The purified enzyme was kinetically characterized with PEA and a novel fluorogenic substrate, N-(4-methyl coumarin) palmitamide (PAMCA). 22040171 6 68 theme residues 1070:1077 arg1 significance 1041:1052 The significance 1037:1052 The significance of the cysteine residues 1037:1077 The significance of the cysteine residues was established by their selective alkylation resulting in almost complete loss of activity. 21987822 4 0 theme first 937:941 arg1 acids 952:956 the highly conserved first 18 amino acids 916:956 the highly conserved first 18 amino acids of the mucin-like domain 916:981 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 4 1 theme Deletion 749:756 arg1 analyses 771:778 Deletion and mutation analyses 749:778 analyses 771:778 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 2 2 theme phosphorylated 514:527 arg1 O-mannose 529:537 ligand-binding moieties to phosphorylated O-mannose 487:537 ligand-binding moieties to phosphorylated O-mannose 487:537 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 7 3 theme extracellular 1676:1688 arg1 proteins 1697:1704 extracellular matrix proteins 1676:1704 extracellular matrix proteins 1676:1704 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 4 4 theme conserved 927:935 arg1 acids 952:956 the highly conserved first 18 amino acids 916:956 the highly conserved first 18 amino acids of the mucin-like domain 916:981 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 1 5 theme O-glycosylated 182:195 arg1 receptor 218:225 a highly O-glycosylated extracellular matrix receptor 173:225 a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells 173:356 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 1 5 theme O-glycosylated 182:195 arg1 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 7 6 theme N-terminal 1378:1387 arg1 region 1389:1394 the N-terminal region 1374:1394 the N-terminal region adjacent to the paired Thr residues 1374:1430 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 0 7 theme laminin 113:119 arg1 binding 121:127 laminin binding 113:127 laminin binding 113:127 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. 21987822 3 8 theme α-dystroglycan 733:746 arg1 α-dystroglycan 733:746 α-dystroglycan 733:746 α-dystroglycan 733:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 3 8 theme α-dystroglycan 733:746 arg1 domain 723:728 the mucin-like domain 708:728 the mucin-like domain of α-dystroglycan 708:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 4 9 theme amino 946:950 arg1 acids 952:956 the highly conserved first 18 amino acids 916:956 the highly conserved first 18 amino acids of the mucin-like domain 916:981 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 7 10 theme Thr 1419:1421 arg1 residues 1423:1430 the paired Thr residues 1408:1430 the paired Thr residues 1408:1430 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 3 11 theme LARGE 580:584 arg1 modification 586:597 the LARGE modification 576:597 the LARGE modification required for laminin- and virus-binding 576:637 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 5 12 theme residues 1015:1022 arg1 importance 988:997 The importance 984:997 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry 984:1107 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 1 13 theme cell 290:293 arg1 surface 295:301 the cell surface 286:301 the cell surface 286:301 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 7 14 theme α-dystroglycan 1597:1610 arg1 terminus 1578:1585 the extreme N terminus 1564:1585 the extreme N terminus of mature α-dystroglycan 1564:1610 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 5 15 theme laminin-binding 1027:1041 arg1 activity 1058:1065 laminin-binding and clustering activity 1027:1065 laminin-binding and clustering activity on myoblasts 1027:1078 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 5 16 from myoblasts 1070:1078 arg1 importance 988:997 The importance 984:997 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry 984:1107 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 7 17 theme ligand-binding 1529:1542 arg1 crucial 1619:1625 crucial 1619:1625 crucial 1619:1625 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 7 17 theme ligand-binding 1529:1542 arg1 activity 1544:1551 the ligand-binding activity 1525:1551 the ligand-binding activity 1525:1551 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 7 18 theme dystroglycan 1482:1493 arg1 maturation 1468:1477 posttranslational maturation 1450:1477 posttranslational maturation of dystroglycan 1450:1493 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 1 19 theme extracellular 197:209 arg1 receptor 218:225 a highly O-glycosylated extracellular matrix receptor 173:225 a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells 173:356 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 1 19 theme extracellular 197:209 arg1 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 3 20 from terminus 696:703 arg1 located 671:677 located 671:677 located 671:677 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 6 21 theme amino 1223:1227 arg1 acids 1229:1233 five amino acids 1218:1233 five amino acids 1218:1233 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 5 22 theme clustering 1047:1056 arg1 activity 1058:1065 laminin-binding and clustering activity 1027:1065 laminin-binding and clustering activity on myoblasts 1027:1078 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 5 23 from activity 1058:1065 arg1 myoblasts 1070:1078 myoblasts 1070:1078 myoblasts 1070:1078 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 1 24 theme matrix 211:216 arg1 receptor 218:225 a highly O-glycosylated extracellular matrix receptor 173:225 a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells 173:356 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 1 24 theme matrix 211:216 arg1 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 7 25 from arenaviruses 1718:1729 arg1 surface 1743:1749 the cell surface 1734:1749 the cell surface 1734:1749 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 5 26 from importance 988:997 arg1 myoblasts 1070:1078 myoblasts 1070:1078 myoblasts 1070:1078 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 5 26 from importance 988:997 arg1 activity 1058:1065 laminin-binding and clustering activity 1027:1065 laminin-binding and clustering activity on myoblasts 1027:1078 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 5 26 from importance 988:997 arg1 entry 1103:1107 arenavirus cell entry 1087:1107 arenavirus cell entry 1087:1107 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 2 27 theme key 407:409 arg1 molecule 411:418 a key molecule 405:418 a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan 405:555 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 2 27 theme key 407:409 arg1 Like-acetylglucosaminyltransferase 359:392 Like-acetylglucosaminyltransferase 359:392 Like-acetylglucosaminyltransferase (LARGE) 359:400 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 2 28 theme α-dystroglycan 459:472 arg1 α-dystroglycan 459:472 α-dystroglycan 459:472 α-dystroglycan 459:472 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 2 28 theme α-dystroglycan 459:472 arg1 domain 449:454 the N-terminal domain 434:454 the N-terminal domain of α-dystroglycan 434:472 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 7 29 theme N 1576:1576 arg1 terminus 1578:1585 the extreme N terminus 1564:1585 the extreme N terminus of mature α-dystroglycan 1564:1610 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 7 30 theme matrix 1690:1695 arg1 proteins 1697:1704 extracellular matrix proteins 1676:1704 extracellular matrix proteins 1676:1704 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 6 31 theme acids 1229:1233 arg1 sequence 1206:1213 a sequence 1204:1213 a sequence 1204:1213 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 7 32 theme adjacent 1396:1403 arg1 region 1389:1394 the N-terminal region 1374:1394 the N-terminal region adjacent to the paired Thr residues 1374:1430 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 6 33 theme ProVal 1255:1260 arg1 sequence 1206:1213 a sequence 1204:1213 a sequence 1204:1213 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 2 34 dep Like-acetylglucosaminyltransferase 359:392 arg1 LARGE 395:399 LARGE 395:399 LARGE 395:399 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 2 35 theme N-terminal 438:447 arg1 α-dystroglycan 459:472 α-dystroglycan 459:472 α-dystroglycan 459:472 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 2 35 theme N-terminal 438:447 arg1 domain 449:454 the N-terminal domain 434:454 the N-terminal domain of α-dystroglycan 434:472 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 6 36 contain contains 1263:1270 arg2 O-glycosylation 1287:1301 phosphorylated O-glycosylation 1272:1301 phosphorylated O-glycosylation 1272:1301 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 6 36 contain contains 1263:1270 arg1 sequence 1206:1213 a sequence 1204:1213 a sequence 1204:1213 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 0 37 theme arenavirus 133:142 arg1 infection 144:152 arenavirus infection 133:152 arenavirus infection 133:152 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. 21987822 3 38 theme N 694:694 arg1 terminus 696:703 the extreme N terminus 682:703 the extreme N terminus of the mucin-like domain of α-dystroglycan 682:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 4 39 theme ligand-binding 801:814 arg1 activity 816:823 the ligand-binding activity 797:823 the ligand-binding activity of α-dystroglycan 797:841 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 5 40 with analysis 1136:1143 arg1 dystroglycan 1162:1173 full-length dystroglycan 1150:1173 full-length dystroglycan 1150:1173 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 7 41 theme paired 1412:1417 arg1 residues 1423:1430 the paired Thr residues 1408:1430 the paired Thr residues 1408:1430 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 6 42 theme phosphorylated 1272:1285 arg1 O-glycosylation 1287:1301 phosphorylated O-glycosylation 1272:1301 phosphorylated O-glycosylation 1272:1301 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 7 43 theme cell 1738:1741 arg1 surface 1743:1749 the cell surface 1734:1749 the cell surface 1734:1749 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 0 44 theme -dependent 42:51 arg1 modification 53:64 Like-acetylglucosaminyltransferase (LARGE)-dependent modification 0:64 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 0:95 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. 21987822 7 45 theme proteins 1697:1704 arg1 assembly 1664:1671 the assembly 1660:1671 the assembly of extracellular matrix proteins 1660:1704 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 2 46 theme moieties 502:509 arg1 O-mannose 529:537 ligand-binding moieties to phosphorylated O-mannose 487:537 ligand-binding moieties to phosphorylated O-mannose 487:537 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 0 47 from Thr-317/319 85:95 arg1 modification 53:64 Like-acetylglucosaminyltransferase (LARGE)-dependent modification 0:64 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 0:95 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. 21987822 3 48 theme specific 649:656 arg1 residues 662:669 specific Thr residues 649:669 specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan 649:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 4 49 from -319 903:906 arg1 modification 875:886 LARGE modification 869:886 LARGE modification at Thr-317 and -319 869:906 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 6 50 dep LARGE 1325:1329 arg1 sufficient 1334:1343 sufficient 1334:1343 sufficient 1334:1343 We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. 21987822 2 51 theme ligand-binding 487:500 arg1 O-mannose 529:537 ligand-binding moieties to phosphorylated O-mannose 487:537 ligand-binding moieties to phosphorylated O-mannose 487:537 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 1 52 theme Old 324:326 arg1 arenaviruses 334:345 Old World arenaviruses 324:345 Old World arenaviruses 324:345 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 2 53 dep O-mannose 529:537 arg1 to 511:512 to 511:512 to 511:512 Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. 21987822 3 54 theme extreme 686:692 arg1 terminus 696:703 the extreme N terminus 682:703 the extreme N terminus of the mucin-like domain of α-dystroglycan 682:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 5 55 theme cell 1098:1101 arg1 entry 1103:1107 arenavirus cell entry 1087:1107 arenavirus cell entry 1087:1107 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 4 56 theme mucin-like 965:974 arg1 domain 976:981 the mucin-like domain 961:981 the mucin-like domain 961:981 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 4 57 theme mutation 762:769 arg1 analyses 771:778 Deletion and mutation analyses 749:778 analyses 771:778 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 1 58 theme World 328:332 arg1 arenaviruses 334:345 Old World arenaviruses 324:345 Old World arenaviruses 324:345 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 3 59 theme mucin-like 712:721 arg1 α-dystroglycan 733:746 α-dystroglycan 733:746 α-dystroglycan 733:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 3 59 theme mucin-like 712:721 arg1 domain 723:728 the mucin-like domain 708:728 the mucin-like domain of α-dystroglycan 708:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 0 60 dep -dependent 42:51 arg1 LARGE 36:40 LARGE 36:40 LARGE 36:40 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. 21987822 4 61 from Thr-317 891:897 arg1 modification 875:886 LARGE modification 869:886 LARGE modification at Thr-317 and -319 869:906 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 5 62 theme full-length 1150:1160 arg1 dystroglycan 1162:1173 full-length dystroglycan 1150:1173 full-length dystroglycan 1150:1173 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 1 63 theme arenaviruses 334:345 arg1 entry 315:319 the entry 311:319 the entry of Old World arenaviruses into cells 311:356 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 1 64 gly O-glycosylated 182:195 arg1 receptor 218:225 a highly O-glycosylated extracellular matrix receptor 173:225 a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells 173:356 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 1 64 gly O-glycosylated 182:195 arg1 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan 155:168 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 3 65 theme domain 723:728 arg1 terminus 696:703 the extreme N terminus 682:703 the extreme N terminus of the mucin-like domain of α-dystroglycan 682:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 3 66 theme located 671:677 arg1 residues 662:669 specific Thr residues 649:669 specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan 649:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 5 67 theme arenavirus 1087:1096 arg1 entry 1103:1107 arenavirus cell entry 1087:1107 arenavirus cell entry 1087:1107 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 5 68 theme paired 1008:1013 arg1 residues 1015:1022 these paired residues 1002:1022 these paired residues 1002:1022 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 7 69 theme posttranslational 1450:1466 arg1 maturation 1468:1477 posttranslational maturation 1450:1477 posttranslational maturation of dystroglycan 1450:1493 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 7 70 theme extreme 1568:1574 arg1 terminus 1578:1585 the extreme N terminus 1564:1585 the extreme N terminus of mature α-dystroglycan 1564:1610 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 4 71 theme α-dystroglycan 828:841 arg1 activity 816:823 the ligand-binding activity 797:823 the ligand-binding activity of α-dystroglycan 797:841 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 4 72 theme domain 976:981 arg1 acids 952:956 the highly conserved first 18 amino acids 916:956 the highly conserved first 18 amino acids of the mucin-like domain 916:981 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 5 73 theme mutational 1125:1134 arg1 analysis 1136:1143 mutational analysis 1125:1143 mutational analysis with full-length dystroglycan 1125:1173 The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. 21987822 3 74 theme Thr 658:660 arg1 residues 662:669 specific Thr residues 649:669 specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan 649:746 Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. 21987822 1 75 theme basement 265:272 arg1 membrane 274:281 the basement membrane 261:281 the basement membrane 261:281 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 21987822 0 76 theme dystroglycan 69:80 arg1 modification 53:64 Like-acetylglucosaminyltransferase (LARGE)-dependent modification 0:64 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 0:95 Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. 21987822 4 77 theme LARGE 869:873 arg1 modification 875:886 LARGE modification 869:886 LARGE modification at Thr-317 and -319 869:906 Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. 21987822 7 78 theme mature 1590:1595 arg1 α-dystroglycan 1597:1610 mature α-dystroglycan 1590:1610 mature α-dystroglycan 1590:1610 Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface. 21987822 1 79 theme membrane 274:281 arg1 anchoring 248:256 anchoring 248:256 anchoring of the basement membrane to the cell surface 248:301 α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. 18930737 10 0 theme salt 1333:1336 arg1 bridges 1338:1344 two salt bridges 1329:1344 two salt bridges 1329:1344 At the two ends of vertical interface, two salt bridges are formed between pairs of Lys41-Asp233 and Lys68-Glu229. 18930737 2 1 theme human 313:317 arg1 plasma 327:332 human seminal plasma 313:332 human seminal plasma 313:332 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 1 2 theme zinc 213:216 arg1 alpha2-glycoprotein 218:236 zinc alpha2-glycoprotein 213:236 zinc alpha2-glycoprotein (ZAG) 213:242 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 1 2 theme zinc 213:216 arg1 ZAG 239:241 ZAG 239:241 ZAG 239:241 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 7 3 theme ZAG 1079:1081 arg1 alpha3 1069:1074 domain alpha3 1062:1074 domain alpha3 of ZAG 1062:1081 The structure of the complex shows that the beta-structure of PIP is ideally aligned with the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins. 18930737 12 4 theme structure 1574:1582 arg1 corner 1551:1556 the corner 1547:1556 the corner of the L-shaped structure 1547:1582 The internal space at the corner of the L-shaped structure is filled with solvent molecules including a carbonate ion. 18930737 7 5 theme long 1093:1096 arg1 interface 1098:1106 a long interface 1091:1106 a long interface between two proteins 1091:1127 The structure of the complex shows that the beta-structure of PIP is ideally aligned with the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins. 18930737 7 6 theme domain 1062:1067 arg1 alpha3 1069:1074 domain alpha3 1062:1074 domain alpha3 of ZAG 1062:1081 The structure of the complex shows that the beta-structure of PIP is ideally aligned with the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins. 18930737 13 7 theme major 1782:1786 arg1 histocompatibility 1788:1805 class I major histocompatibility 1774:1805 the class I major histocompatibility complex structures 1770:1824 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 7 8 theme complex 971:977 arg1 structure 954:962 The structure 950:962 The structure of the complex 950:977 The structure of the complex shows that the beta-structure of PIP is ideally aligned with the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins. 18930737 12 9 theme L-shaped 1565:1572 arg1 structure 1574:1582 the L-shaped structure 1561:1582 the L-shaped structure 1561:1582 The internal space at the corner of the L-shaped structure is filled with solvent molecules including a carbonate ion. 18930737 6 10 dep found 905:909 arg1 glycosylated 911:922 glycosylated 911:922 is found glycosylated with four sugar residues 902:947 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 7 11 theme alpha3 1069:1074 arg1 beta-structure 1044:1057 the beta-structure 1040:1057 the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins 1040:1127 The structure of the complex shows that the beta-structure of PIP is ideally aligned with the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins. 18930737 13 12 theme histocompatibility 1788:1805 arg1 structures 1815:1824 the class I major histocompatibility complex structures 1770:1824 the class I major histocompatibility complex structures 1770:1824 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 6 13 theme N-glycosylation 858:872 arg1 site 874:877 one potential N-glycosylation site 844:877 one potential N-glycosylation site at Asn77 844:886 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 0 14 gly alpha2-glycoprotein 59:77 arg1 ZAG 80:82 ZAG 80:82 ZAG 80:82 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 0 14 gly alpha2-glycoprotein 59:77 arg1 alpha2-glycoprotein 59:77 zinc alpha2-glycoprotein 54:77 zinc alpha2-glycoprotein (ZAG) 54:83 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 13 15 from area 1663:1666 arg1 complex 1675:1681 the complex 1671:1681 the complex 1671:1681 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 5 16 theme sheets 741:746 arg1 form 703:706 the form 699:706 the form of two antiparallel beta-pleated sheets 699:746 The scaffolding of PIP consists of seven beta-strands that are organized in the form of two antiparallel beta-pleated sheets, resulting in the formation of a sandwiched beta-sheet. 18930737 10 17 theme Lys68-Glu229 1391:1402 arg1 pairs 1365:1369 pairs 1365:1369 pairs of Lys41-Asp233 and Lys68-Glu229 1365:1402 At the two ends of vertical interface, two salt bridges are formed between pairs of Lys41-Asp233 and Lys68-Glu229. 18930737 11 18 theme salt 1502:1505 arg1 bridge 1507:1512 another salt bridge 1494:1512 another salt bridge 1494:1512 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 8 19 theme proximal 1134:1141 arg1 beta-strands 1143:1154 The proximal beta-strands 1130:1154 The proximal beta-strands at the long interface 1130:1176 The proximal beta-strands at the long interface are arranged in an antiparallel manner. 18930737 3 20 theme crystallographic 475:490 arg1 method 492:497 X-ray crystallographic method 469:497 X-ray crystallographic method 469:497 The structure of the complex has been determined using X-ray crystallographic method and refined to an R(cryst) of 0.199 (R(free)=0.239). 18930737 6 21 theme sugar 934:938 arg1 residues 940:947 four sugar residues 929:947 four sugar residues 929:947 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 2 22 theme 20 357:358 arg1 % 359:359 % 359:359 % 359:359 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 0 23 theme prolactin-inducible 89:107 arg1 PIP 118:120 PIP 118:120 PIP 118:120 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 0 23 theme prolactin-inducible 89:107 arg1 protein 109:115 prolactin-inducible protein 89:115 prolactin-inducible protein (PIP) 89:121 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 11 24 theme PIP 1489:1491 arg1 loop 1481:1484 a loop 1479:1484 a loop of PIP 1479:1491 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 24 theme PIP 1489:1491 arg1 domains 1460:1466 alpha1-alpha2 domains 1446:1466 alpha1-alpha2 domains of ZAG 1446:1473 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 24 theme PIP 1489:1491 arg1 PIP 1489:1491 PIP 1489:1491 PIP 1489:1491 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 24 theme PIP 1489:1491 arg1 ZAG 1471:1473 ZAG 1471:1473 ZAG 1471:1473 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 7 25 theme PIP 1012:1014 arg1 beta-structure 994:1007 the beta-structure 990:1007 the beta-structure of PIP 990:1014 The structure of the complex shows that the beta-structure of PIP is ideally aligned with the beta-structure of domain alpha3 of ZAG to form a long interface between two proteins. 18930737 1 26 gly alpha2-glycoprotein 218:236 arg1 alpha2-glycoprotein 218:236 zinc alpha2-glycoprotein 213:236 zinc alpha2-glycoprotein (ZAG) 213:242 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 1 26 gly alpha2-glycoprotein 218:236 arg1 ZAG 239:241 ZAG 239:241 ZAG 239:241 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 0 27 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the novel complex 0:37 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 2 28 theme hexaethylene 393:404 arg1 glycol 406:411 5% hexaethylene glycol 390:411 5% hexaethylene glycol 390:411 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 6 29 from Asn77 882:886 arg1 site 874:877 one potential N-glycosylation site 844:877 one potential N-glycosylation site at Asn77 844:886 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 11 30 theme perpendicular 1412:1424 arg1 interface 1426:1434 the perpendicular interface 1408:1434 the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP 1408:1491 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 4 31 theme ZAG 569:571 arg1 similar 584:590 similar 584:590 similar 584:590 The structure of ZAG is broadly similar to the structure of serum ZAG. 18930737 4 31 theme ZAG 569:571 arg1 structure 556:564 The structure 552:564 The structure of ZAG 552:571 The structure of ZAG is broadly similar to the structure of serum ZAG. 18930737 2 32 theme % 391:391 arg1 glycol 406:411 5% hexaethylene glycol 390:411 5% hexaethylene glycol 390:411 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 6 33 theme potential 848:856 arg1 site 874:877 one potential N-glycosylation site 844:877 one potential N-glycosylation site at Asn77 844:886 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 8 34 from interface 1168:1176 arg1 beta-strands 1143:1154 The proximal beta-strands 1130:1154 The proximal beta-strands at the long interface 1130:1176 The proximal beta-strands at the long interface are arranged in an antiparallel manner. 18930737 4 35 theme ZAG 618:620 arg1 structure 599:607 the structure 595:607 the structure of serum ZAG 595:620 The structure of ZAG is broadly similar to the structure of serum ZAG. 18930737 1 36 theme first 162:166 arg1 This 150:153 This 150:153 This 150:153 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 1 36 theme first 162:166 arg1 report 168:173 the first report 158:173 the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) 158:280 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 2 37 theme 5 390:390 arg1 % 391:391 % 391:391 % 391:391 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 6 38 gly glycosylated 911:922 arg1 same 897:900 same 897:900 same 897:900 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 13 39 theme overall 1648:1654 arg1 A 1704:1704 approximately 914 A 1686:1704 approximately 914 A(2) 1686:1707 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 13 39 theme overall 1648:1654 arg1 area 1663:1666 The overall buried area 1644:1666 The overall buried area in the complex 1644:1681 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 1 40 theme prolactin-inducible 248:266 arg1 PIP 277:279 PIP 277:279 PIP 277:279 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 1 40 theme prolactin-inducible 248:266 arg1 protein 268:274 prolactin-inducible protein 248:274 prolactin-inducible protein (PIP) 248:280 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 6 41 gly N-glycosylation 858:872 arg2 one 844:846 one 844:846 one 844:846 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 6 41 gly N-glycosylation 858:872 arg2 Asn77 882:886 Asn77 882:886 Asn77 882:886 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 6 41 gly N-glycosylation 858:872 arg2 site 874:877 one potential N-glycosylation site 844:877 one potential N-glycosylation site at Asn77 844:886 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 13 42 theme buried 1656:1661 arg1 A 1704:1704 approximately 914 A 1686:1704 approximately 914 A(2) 1686:1707 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 13 42 theme buried 1656:1661 arg1 area 1663:1666 The overall buried area 1644:1666 The overall buried area in the complex 1644:1681 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 3 43 dep R 536:536 arg1 free 538:541 free 538:541 free 538:541 The structure of the complex has been determined using X-ray crystallographic method and refined to an R(cryst) of 0.199 (R(free)=0.239). 18930737 0 44 theme complex 31:37 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the novel complex 0:37 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 6 45 theme PIP 831:833 arg1 sequence 819:826 The amino acid sequence 804:826 The amino acid sequence of PIP 804:833 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 10 46 theme interface 1318:1326 arg1 ends 1301:1304 the two ends 1293:1304 the two ends of vertical interface 1293:1326 At the two ends of vertical interface, two salt bridges are formed between pairs of Lys41-Asp233 and Lys68-Glu229. 18930737 5 47 theme antiparallel 715:726 arg1 sheets 741:746 two antiparallel beta-pleated sheets 711:746 two antiparallel beta-pleated sheets 711:746 The scaffolding of PIP consists of seven beta-strands that are organized in the form of two antiparallel beta-pleated sheets, resulting in the formation of a sandwiched beta-sheet. 18930737 2 48 theme polyethylene 361:372 arg1 glycol 374:379 20% polyethylene glycol 9000 357:384 20% polyethylene glycol 9000 357:384 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 0 49 theme novel 25:29 arg1 complex 31:37 the novel complex 21:37 the novel complex 21:37 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 10 50 theme vertical 1309:1316 arg1 interface 1318:1326 vertical interface 1309:1326 vertical interface 1309:1326 At the two ends of vertical interface, two salt bridges are formed between pairs of Lys41-Asp233 and Lys68-Glu229. 18930737 8 51 theme long 1163:1166 arg1 interface 1168:1176 the long interface 1159:1176 the long interface 1159:1176 The proximal beta-strands at the long interface are arranged in an antiparallel manner. 18930737 3 52 theme 0.199 529:533 arg1 cryst 519:523 cryst 519:523 cryst 519:523 The structure of the complex has been determined using X-ray crystallographic method and refined to an R(cryst) of 0.199 (R(free)=0.239). 18930737 3 52 theme 0.199 529:533 arg1 R 517:517 an R 514:517 an R(cryst) of 0.199 (R(free)=0.239) 514:549 The structure of the complex has been determined using X-ray crystallographic method and refined to an R(cryst) of 0.199 (R(free)=0.239). 18930737 9 53 theme salt 1256:1259 arg1 bridges 1261:1267 three salt bridges 1250:1267 three salt bridges 1250:1267 There are 12 hydrogen bonds and three salt bridges between ZAG and PIP. 18930737 2 54 theme % 359:359 arg1 glycol 374:379 20% polyethylene glycol 9000 357:384 20% polyethylene glycol 9000 357:384 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 18930737 13 55 theme complex 1807:1813 arg1 structures 1815:1824 the class I major histocompatibility complex structures 1770:1824 the class I major histocompatibility complex structures 1770:1824 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 0 56 theme seminal 134:140 arg1 plasma 142:147 human seminal plasma 128:147 human seminal plasma 128:147 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 5 57 theme PIP 642:644 arg1 scaffolding 627:637 The scaffolding 623:637 The scaffolding of PIP 623:644 The scaffolding of PIP consists of seven beta-strands that are organized in the form of two antiparallel beta-pleated sheets, resulting in the formation of a sandwiched beta-sheet. 18930737 13 58 theme class 1774:1778 arg1 histocompatibility 1788:1805 class I major histocompatibility 1774:1805 the class I major histocompatibility complex structures 1770:1824 The overall buried area in the complex is approximately 914 A(2), which is considerably higher than the 660 A(2) reported for the class I major histocompatibility complex structures. 18930737 3 59 theme complex 435:441 arg1 structure 418:426 The structure 414:426 The structure of the complex 414:441 The structure of the complex has been determined using X-ray crystallographic method and refined to an R(cryst) of 0.199 (R(free)=0.239). 18930737 6 60 theme acid 814:817 arg1 sequence 819:826 The amino acid sequence 804:826 The amino acid sequence of PIP 804:833 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 0 61 theme human 128:132 arg1 plasma 142:147 human seminal plasma 128:147 human seminal plasma 128:147 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 1 62 from report 168:173 arg1 formation 182:190 the formation 178:190 the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) 178:280 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 5 63 theme beta-sheet 792:801 arg1 formation 766:774 the formation 762:774 the formation of a sandwiched beta-sheet 762:801 The scaffolding of PIP consists of seven beta-strands that are organized in the form of two antiparallel beta-pleated sheets, resulting in the formation of a sandwiched beta-sheet. 18930737 12 64 theme internal 1529:1536 arg1 space 1538:1542 The internal space 1525:1542 The internal space at the corner of the L-shaped structure 1525:1582 The internal space at the corner of the L-shaped structure is filled with solvent molecules including a carbonate ion. 18930737 6 65 theme amino 808:812 arg1 sequence 819:826 The amino acid sequence 804:826 The amino acid sequence of PIP 804:833 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 6 66 contain contains 835:842 arg1 sequence 819:826 The amino acid sequence 804:826 The amino acid sequence of PIP 804:833 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 6 66 contain contains 835:842 arg2 site 874:877 one potential N-glycosylation site 844:877 one potential N-glycosylation site at Asn77 844:886 The amino acid sequence of PIP contains one potential N-glycosylation site at Asn77, and the same is found glycosylated with four sugar residues. 18930737 4 67 theme serum 612:616 arg1 ZAG 618:620 serum ZAG 612:620 serum ZAG 612:620 The structure of ZAG is broadly similar to the structure of serum ZAG. 18930737 9 68 theme hydrogen 1231:1238 arg1 bonds 1240:1244 12 hydrogen bonds 1228:1244 12 hydrogen bonds 1228:1244 There are 12 hydrogen bonds and three salt bridges between ZAG and PIP. 18930737 8 69 theme antiparallel 1197:1208 arg1 manner 1210:1215 an antiparallel manner 1194:1215 an antiparallel manner 1194:1215 The proximal beta-strands at the long interface are arranged in an antiparallel manner. 18930737 12 70 from corner 1551:1556 arg1 space 1538:1542 The internal space 1525:1542 The internal space at the corner of the L-shaped structure 1525:1582 The internal space at the corner of the L-shaped structure is filled with solvent molecules including a carbonate ion. 18930737 12 71 theme carbonate 1629:1637 arg1 ion 1639:1641 a carbonate ion 1627:1641 a carbonate ion 1627:1641 The internal space at the corner of the L-shaped structure is filled with solvent molecules including a carbonate ion. 18930737 12 72 theme solvent 1599:1605 arg1 molecules 1607:1615 solvent molecules 1599:1615 solvent molecules including a carbonate ion 1599:1641 The internal space at the corner of the L-shaped structure is filled with solvent molecules including a carbonate ion. 18930737 3 73 theme X-ray 469:473 arg1 method 492:497 X-ray crystallographic method 469:497 X-ray crystallographic method 469:497 The structure of the complex has been determined using X-ray crystallographic method and refined to an R(cryst) of 0.199 (R(free)=0.239). 18930737 10 74 theme Lys41-Asp233 1374:1385 arg1 pairs 1365:1369 pairs 1365:1369 pairs of Lys41-Asp233 and Lys68-Glu229 1365:1402 At the two ends of vertical interface, two salt bridges are formed between pairs of Lys41-Asp233 and Lys68-Glu229. 18930737 5 75 theme beta-pleated 728:739 arg1 sheets 741:746 two antiparallel beta-pleated sheets 711:746 two antiparallel beta-pleated sheets 711:746 The scaffolding of PIP consists of seven beta-strands that are organized in the form of two antiparallel beta-pleated sheets, resulting in the formation of a sandwiched beta-sheet. 18930737 11 76 theme ZAG 1471:1473 arg1 loop 1481:1484 a loop 1479:1484 a loop of PIP 1479:1491 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 76 theme ZAG 1471:1473 arg1 domains 1460:1466 alpha1-alpha2 domains 1446:1466 alpha1-alpha2 domains of ZAG 1446:1473 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 76 theme ZAG 1471:1473 arg1 PIP 1489:1491 PIP 1489:1491 PIP 1489:1491 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 76 theme ZAG 1471:1473 arg1 ZAG 1471:1473 ZAG 1471:1473 ZAG 1471:1473 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 1 77 theme complex 197:203 arg1 formation 182:190 the formation 178:190 the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) 178:280 This is the first report on the formation of a complex between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP). 18930737 0 78 theme zinc 54:57 arg1 ZAG 80:82 ZAG 80:82 ZAG 80:82 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 0 78 theme zinc 54:57 arg1 alpha2-glycoprotein 59:77 zinc alpha2-glycoprotein 54:77 zinc alpha2-glycoprotein (ZAG) 54:83 Crystal structure of the novel complex formed between zinc alpha2-glycoprotein (ZAG) and prolactin-inducible protein (PIP) from human seminal plasma. 18930737 11 79 theme alpha1-alpha2 1446:1458 arg1 domains 1460:1466 alpha1-alpha2 domains 1446:1466 alpha1-alpha2 domains of ZAG 1446:1473 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 79 theme alpha1-alpha2 1446:1458 arg1 PIP 1489:1491 PIP 1489:1491 PIP 1489:1491 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 11 79 theme alpha1-alpha2 1446:1458 arg1 ZAG 1471:1473 ZAG 1471:1473 ZAG 1471:1473 On the perpendicular interface involving alpha1-alpha2 domains of ZAG and a loop of PIP, another salt bridge is formed. 18930737 2 80 theme seminal 319:325 arg1 plasma 327:332 human seminal plasma 313:332 human seminal plasma 313:332 The complex was purified from human seminal plasma and crystallized using 20% polyethylene glycol 9000 and 5% hexaethylene glycol. 10066782 7 0 theme FN 1101:1102 arg1 domains 1108:1114 the Ig-like and NH2-terminal FN III domains 1072:1114 the Ig-like and NH2-terminal FN III domains causing them to fold 1072:1135 An unusual cysteine connectivity between Cys6-Cys174, which links the Ig-like and NH2-terminal FN III domains causing them to fold back onto each other, has not previously been observed among cytokine receptors. 10066782 6 1 theme domain 940:945 arg1 motifs 947:952 known cytokine-binding domain motifs 917:952 known cytokine-binding domain motifs 917:952 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 1 2 theme IL-6 229:232 arg1 receptor 234:241 IL-6 receptor 229:241 IL-6 receptor (IL-6R) 229:249 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 2 theme IL-6 229:232 arg1 IL-6R 244:248 IL-6R 244:248 IL-6R 244:248 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 2 3 theme Ig-like 415:421 arg1 domain 423:428 an amino-terminal Ig-like domain 397:428 an amino-terminal Ig-like domain 397:428 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 3 4 theme WSXWS 593:597 arg1 motif 608:612 a WSXWS sequence motif 591:612 a WSXWS sequence motif 591:612 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 2 5 dep domains 388:394 arg1 domains 468:474 two fibronectin-type III (FN III) domains 434:474 two fibronectin-type III (FN III) domains 434:474 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 2 5 dep domains 388:394 arg1 domain 423:428 an amino-terminal Ig-like domain 397:428 an amino-terminal Ig-like domain 397:428 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 5 6 theme free 834:837 arg1 cysteines 839:847 two free cysteines 830:847 two free cysteines 830:847 Mass spectrometric analysis of these peptides revealed four disulfide bonds and two free cysteines. 10066782 0 7 theme interleukin-6 92:104 arg1 receptor 106:113 the human interleukin-6 receptor 82:113 the human interleukin-6 receptor 82:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 3 8 theme sequence 599:606 arg1 motif 608:612 a WSXWS sequence motif 591:612 a WSXWS sequence motif 591:612 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 8 9 theme Cys258 1330:1335 arg1 proportion 1316:1325 a small proportion 1308:1325 a small proportion of Cys258 1308:1335 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 9 theme Cys258 1330:1335 arg1 reactive 1341:1348 reactive 1341:1348 reactive 1341:1348 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 5 10 theme peptides 787:794 arg1 analysis 769:776 Mass spectrometric analysis 750:776 Mass spectrometric analysis of these peptides 750:794 Mass spectrometric analysis of these peptides revealed four disulfide bonds and two free cysteines. 10066782 0 11 theme human 86:90 arg1 receptor 106:113 the human interleukin-6 receptor 82:113 the human interleukin-6 receptor 82:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 7 12 theme Ig-like 1076:1082 arg1 domains 1108:1114 the Ig-like and NH2-terminal FN III domains 1072:1114 the Ig-like and NH2-terminal FN III domains causing them to fold 1072:1135 An unusual cysteine connectivity between Cys6-Cys174, which links the Ig-like and NH2-terminal FN III domains causing them to fold back onto each other, has not previously been observed among cytokine receptors. 10066782 4 13 theme reducing 734:741 arg1 agent 743:747 reducing agent 734:747 reducing agent 734:747 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 4 14 theme sIL-6R 677:682 arg1 structure 654:662 the disulfide structure 640:662 the disulfide structure of the human sIL-6R 640:682 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 2 15 theme " 341:341 arg1 part 343:346 The extracellular "soluble" part 315:346 The extracellular "soluble" part of the IL-6R (sIL-6R) 315:368 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 5 16 theme Mass 750:753 arg1 analysis 769:776 Mass spectrometric analysis 750:776 Mass spectrometric analysis of these peptides 750:794 Mass spectrometric analysis of these peptides revealed four disulfide bonds and two free cysteines. 10066782 0 17 theme receptor 106:113 arg1 receptor 106:113 the human interleukin-6 receptor 82:113 the human interleukin-6 receptor 82:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 17 theme receptor 106:113 arg1 domain 72:77 the extracellular domain 54:77 the extracellular domain of the human interleukin-6 receptor 54:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 6 18 theme known 917:921 arg1 motifs 947:952 known cytokine-binding domain motifs 917:952 known cytokine-binding domain motifs 917:952 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 4 19 theme human 671:675 arg1 sIL-6R 677:682 the human sIL-6R 667:682 the human sIL-6R 667:682 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 5 20 theme spectrometric 755:767 arg1 analysis 769:776 Mass spectrometric analysis 750:776 Mass spectrometric analysis of these peptides 750:794 Mass spectrometric analysis of these peptides revealed four disulfide bonds and two free cysteines. 10066782 0 21 theme bond 10:13 arg1 structure 15:23 Disulfide bond structure 0:23 Disulfide bond structure 0:23 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 3 22 theme FN 485:486 arg1 domains 492:498 The two FN III domains 477:498 The two FN III domains 477:498 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 6 23 theme superfamily 985:995 arg1 domains 997:1003 known Ig superfamily domains 976:1003 known Ig superfamily domains 976:1003 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 1 24 theme hexameric 169:177 arg1 complex 179:185 a hexameric complex 167:185 a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130 167:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 24 theme hexameric 169:177 arg1 receptor 155:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 0 25 theme Disulfide 0:8 arg1 structure 15:23 Disulfide bond structure 0:23 Disulfide bond structure 0:23 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 4 26 theme agent 743:747 arg1 absence 710:716 absence 710:716 absence 710:716 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 4 26 theme agent 743:747 arg1 presence 722:729 presence 722:729 presence 722:729 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 7 27 theme cysteine 1017:1024 arg1 connectivity 1026:1037 An unusual cysteine connectivity 1006:1037 An unusual cysteine connectivity 1006:1037 An unusual cysteine connectivity between Cys6-Cys174, which links the Ig-like and NH2-terminal FN III domains causing them to fold back onto each other, has not previously been observed among cytokine receptors. 10066782 2 28 theme soluble 334:340 arg1 part 343:346 The extracellular "soluble" part 315:346 The extracellular "soluble" part of the IL-6R (sIL-6R) 315:368 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 7 29 theme NH2-terminal 1088:1099 arg1 domains 1108:1114 the Ig-like and NH2-terminal FN III domains 1072:1114 the Ig-like and NH2-terminal FN III domains causing them to fold 1072:1135 An unusual cysteine connectivity between Cys6-Cys174, which links the Ig-like and NH2-terminal FN III domains causing them to fold back onto each other, has not previously been observed among cytokine receptors. 10066782 8 30 with reactive 1341:1348 arg1 4-vinylpyridine 1376:1390 the alkylating agent 4-vinylpyridine 1355:1390 the alkylating agent 4-vinylpyridine 1355:1390 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 31 theme small 1310:1314 arg1 proportion 1316:1325 a small proportion 1308:1325 a small proportion of Cys258 1308:1335 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 31 theme small 1310:1314 arg1 reactive 1341:1348 reactive 1341:1348 reactive 1341:1348 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 32 theme free 1226:1229 arg1 cysteines 1231:1239 The two free cysteines 1218:1239 The two free cysteines (Cys192 and Cys258) 1218:1259 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 32 theme free 1226:1229 arg1 cysteinyl-cysteines 1278:1296 cysteinyl-cysteines 1278:1296 cysteinyl-cysteines 1278:1296 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 32 theme free 1226:1229 arg1 Cys258 1253:1258 Cys258 1253:1258 Cys258 1253:1258 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 32 theme free 1226:1229 arg1 Cys192 1242:1247 Cys192 1242:1247 Cys192 1242:1247 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 6 33 theme known 976:980 arg1 domains 997:1003 known Ig superfamily domains 976:1003 known Ig superfamily domains 976:1003 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 6 34 dep disulfides 854:863 arg1 Cys102-Cys113 865:877 Cys102-Cys113 865:877 Cys102-Cys113 865:877 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 6 34 dep disulfides 854:863 arg1 disulfides 854:863 The disulfides Cys102-Cys113 and Cys146-Cys157 850:895 The disulfides Cys102-Cys113 and Cys146-Cys157 850:895 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 6 34 dep disulfides 854:863 arg1 Cys146-Cys157 883:895 Cys146-Cys157 883:895 Cys146-Cys157 883:895 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 9 35 theme N-glycosylation 1415:1429 arg1 sites 1431:1435 the four potential N-glycosylation sites 1396:1435 the four potential N-glycosylation sites 1396:1435 Of the four potential N-glycosylation sites, carbohydrate moieties were identified on Asn36, Asn74, and Asn202, but not on Asn226. 10066782 4 36 theme disulfide 644:652 arg1 structure 654:662 the disulfide structure 640:662 the disulfide structure of the human sIL-6R 640:682 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 2 37 theme amino-terminal 400:413 arg1 domain 423:428 an amino-terminal Ig-like domain 397:428 an amino-terminal Ig-like domain 397:428 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 7 38 theme cytokine 1198:1205 arg1 receptors 1207:1215 cytokine receptors 1198:1215 cytokine receptors 1198:1215 An unusual cysteine connectivity between Cys6-Cys174, which links the Ig-like and NH2-terminal FN III domains causing them to fold back onto each other, has not previously been observed among cytokine receptors. 10066782 3 39 theme conserved 559:567 arg1 residues 578:585 4 conserved cysteine residues 557:585 4 conserved cysteine residues 557:585 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 6 40 theme Ig 982:983 arg1 domains 997:1003 known Ig superfamily domains 976:1003 known Ig superfamily domains 976:1003 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 2 41 theme IL-6R 355:359 arg1 part 343:346 The extracellular "soluble" part 315:346 The extracellular "soluble" part of the IL-6R (sIL-6R) 315:368 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 0 42 theme N-glycosylation 29:43 arg1 sites 45:49 N-glycosylation sites 29:49 N-glycosylation sites 29:49 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 42 theme N-glycosylation 29:43 arg1 domain 72:77 the extracellular domain 54:77 the extracellular domain of the human interleukin-6 receptor 54:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 1 43 theme high 260:263 arg1 gp130 308:312 gp130 308:312 gp130 308:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 43 theme high 260:263 arg1 converter 274:282 the high affinity converter 256:282 the high affinity converter 256:282 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 8 44 theme agent 1370:1374 arg1 4-vinylpyridine 1376:1390 the alkylating agent 4-vinylpyridine 1355:1390 the alkylating agent 4-vinylpyridine 1355:1390 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 3 45 theme cytokine-binding 513:528 arg1 domain 530:535 the cytokine-binding domain 509:535 the cytokine-binding domain defined by a set of 4 conserved cysteine residues 509:585 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 2 46 theme fibronectin-type 438:453 arg1 domains 468:474 two fibronectin-type III (FN III) domains 434:474 two fibronectin-type III (FN III) domains 434:474 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 2 47 theme FN 460:461 arg1 domains 468:474 two fibronectin-type III (FN III) domains 434:474 two fibronectin-type III (FN III) domains 434:474 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 8 48 theme alkylating 1359:1368 arg1 4-vinylpyridine 1376:1390 the alkylating agent 4-vinylpyridine 1355:1390 the alkylating agent 4-vinylpyridine 1355:1390 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 49 dep cysteines 1231:1239 arg1 cysteines 1231:1239 The two free cysteines 1218:1239 The two free cysteines (Cys192 and Cys258) 1218:1259 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 49 dep cysteines 1231:1239 arg1 Cys258 1253:1258 Cys258 1253:1258 Cys258 1253:1258 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 8 49 dep cysteines 1231:1239 arg1 Cys192 1242:1247 Cys192 1242:1247 Cys192 1242:1247 The two free cysteines (Cys192 and Cys258) were detected as cysteinyl-cysteines, although a small proportion of Cys258 was reactive with the alkylating agent 4-vinylpyridine. 10066782 5 50 theme disulfide 810:818 arg1 bonds 820:824 four disulfide bonds 805:824 four disulfide bonds 805:824 Mass spectrometric analysis of these peptides revealed four disulfide bonds and two free cysteines. 10066782 2 51 theme extracellular 319:331 arg1 part 343:346 The extracellular "soluble" part 315:346 The extracellular "soluble" part of the IL-6R (sIL-6R) 315:368 The extracellular "soluble" part of the IL-6R (sIL-6R) consists of three domains: an amino-terminal Ig-like domain and two fibronectin-type III (FN III) domains. 10066782 9 52 gly N-glycosylation 1415:1429 arg2 four 1400:1403 four 1400:1403 four 1400:1403 Of the four potential N-glycosylation sites, carbohydrate moieties were identified on Asn36, Asn74, and Asn202, but not on Asn226. 10066782 9 52 gly N-glycosylation 1415:1429 arg2 sites 1431:1435 the four potential N-glycosylation sites 1396:1435 the four potential N-glycosylation sites 1396:1435 Of the four potential N-glycosylation sites, carbohydrate moieties were identified on Asn36, Asn74, and Asn202, but not on Asn226. 10066782 6 53 with consistent 901:910 arg1 motifs 947:952 known cytokine-binding domain motifs 917:952 known cytokine-binding domain motifs 917:952 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 6 53 with consistent 901:910 arg1 Cys28-Cys77 959:969 Cys28-Cys77 959:969 Cys28-Cys77 959:969 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 6 54 with motifs 947:952 arg1 domains 997:1003 known Ig superfamily domains 976:1003 known Ig superfamily domains 976:1003 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 1 55 theme high 120:123 arg1 complex 179:185 a hexameric complex 167:185 a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130 167:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 55 theme high 120:123 arg1 receptor 155:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 4 56 theme peptide 687:693 arg1 mapping 695:701 peptide mapping 687:701 peptide mapping 687:701 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 0 57 gly N-glycosylation 29:43 arg2 sites 45:49 N-glycosylation sites 29:49 N-glycosylation sites 29:49 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 57 gly N-glycosylation 29:43 arg1 receptor 106:113 the human interleukin-6 receptor 82:113 the human interleukin-6 receptor 82:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 57 gly N-glycosylation 29:43 arg1 domain 72:77 the extracellular domain 54:77 the extracellular domain of the human interleukin-6 receptor 54:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 57 gly N-glycosylation 29:43 arg2 domain 72:77 the extracellular domain 54:77 the extracellular domain of the human interleukin-6 receptor 54:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 9 58 theme carbohydrate 1438:1449 arg1 moieties 1451:1458 carbohydrate moieties 1438:1458 carbohydrate moieties 1438:1458 Of the four potential N-glycosylation sites, carbohydrate moieties were identified on Asn36, Asn74, and Asn202, but not on Asn226. 10066782 1 59 theme affinity 125:132 arg1 complex 179:185 a hexameric complex 167:185 a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130 167:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 59 theme affinity 125:132 arg1 receptor 155:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 3 60 theme cysteine 569:576 arg1 residues 578:585 4 conserved cysteine residues 557:585 4 conserved cysteine residues 557:585 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 1 61 dep molecules 205:213 arg1 each 215:218 each 215:218 each 215:218 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 61 dep molecules 205:213 arg1 converter 274:282 the high affinity converter 256:282 the high affinity converter 256:282 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 61 dep molecules 205:213 arg1 IL-6 223:226 IL-6 223:226 IL-6 223:226 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 61 dep molecules 205:213 arg1 receptor 234:241 IL-6 receptor 229:241 IL-6 receptor (IL-6R) 229:249 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 61 dep molecules 205:213 arg1 gp130 308:312 gp130 308:312 gp130 308:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 61 dep molecules 205:213 arg1 IL-6R 244:248 IL-6R 244:248 IL-6R 244:248 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 62 theme interleukin-6 134:146 arg1 complex 179:185 a hexameric complex 167:185 a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130 167:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 62 theme interleukin-6 134:146 arg1 receptor 155:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 7 63 theme unusual 1009:1015 arg1 connectivity 1026:1037 An unusual cysteine connectivity 1006:1037 An unusual cysteine connectivity 1006:1037 An unusual cysteine connectivity between Cys6-Cys174, which links the Ig-like and NH2-terminal FN III domains causing them to fold back onto each other, has not previously been observed among cytokine receptors. 10066782 0 64 theme domain 72:77 arg1 structure 15:23 Disulfide bond structure 0:23 Disulfide bond structure 0:23 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 64 theme domain 72:77 arg1 sites 45:49 N-glycosylation sites 29:49 N-glycosylation sites 29:49 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 64 theme domain 72:77 arg1 domain 72:77 the extracellular domain 54:77 the extracellular domain of the human interleukin-6 receptor 54:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 3 65 theme residues 578:585 arg1 residues 578:585 4 conserved cysteine residues 557:585 4 conserved cysteine residues 557:585 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 3 65 theme residues 578:585 arg1 set 550:552 a set 548:552 a set of 4 conserved cysteine residues 548:585 The two FN III domains comprise the cytokine-binding domain defined by a set of 4 conserved cysteine residues and a WSXWS sequence motif. 10066782 9 66 theme potential 1405:1413 arg1 sites 1431:1435 the four potential N-glycosylation sites 1396:1435 the four potential N-glycosylation sites 1396:1435 Of the four potential N-glycosylation sites, carbohydrate moieties were identified on Asn36, Asn74, and Asn202, but not on Asn226. 10066782 4 67 dep absence 710:716 arg1 the 706:708 the 706:708 the 706:708 Here, we have determined the disulfide structure of the human sIL-6R by peptide mapping in the absence and presence of reducing agent. 10066782 0 68 theme extracellular 58:70 arg1 receptor 106:113 the human interleukin-6 receptor 82:113 the human interleukin-6 receptor 82:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 0 68 theme extracellular 58:70 arg1 domain 72:77 the extracellular domain 54:77 the extracellular domain of the human interleukin-6 receptor 54:113 Disulfide bond structure and N-glycosylation sites of the extracellular domain of the human interleukin-6 receptor. 10066782 1 69 theme signaling 288:296 arg1 molecule 298:305 signaling molecule 288:305 signaling molecule 288:305 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 6 70 theme cytokine-binding 923:938 arg1 motifs 947:952 known cytokine-binding domain motifs 917:952 known cytokine-binding domain motifs 917:952 The disulfides Cys102-Cys113 and Cys146-Cys157 are consistent with known cytokine-binding domain motifs, and Cys28-Cys77 with known Ig superfamily domains. 10066782 1 71 theme IL-6 149:152 arg1 complex 179:185 a hexameric complex 167:185 a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130 167:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 71 theme IL-6 149:152 arg1 receptor 155:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor 116:162 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 72 theme affinity 265:272 arg1 gp130 308:312 gp130 308:312 gp130 308:312 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 10066782 1 72 theme affinity 265:272 arg1 converter 274:282 the high affinity converter 256:282 the high affinity converter 256:282 The high affinity interleukin-6 (IL-6) receptor is a hexameric complex consisting of two molecules each of IL-6, IL-6 receptor (IL-6R), and the high affinity converter and signaling molecule, gp130. 22735700 5 0 theme 1.6 Å 689:693 arg1 resolution 695:704 1.6 Å resolution 689:704 1.6 Å resolution 689:704 We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. 22735700 6 1 theme structural 858:867 arg1 differences 869:879 distinct structural differences 849:879 distinct structural differences 849:879 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 7 2 theme catalytic 886:894 arg1 features 896:903 The catalytic features 882:903 The catalytic features of RNase T2 in presence of bivalent cations 882:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 1 3 theme matter 117:122 arg1 disease 124:130 white matter disease 111:130 white matter disease of the human brain 111:149 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 3 4 from humans 479:484 arg1 family 448:453 the Rh/T2/S family 436:453 the Rh/T2/S family of acidic hydrolases in humans 436:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 3 5 from family 448:453 arg1 humans 479:484 humans 479:484 humans 479:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 4 6 theme activity 624:631 arg1 independent 595:605 independent 595:605 independent 595:605 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 7 7 theme mutations 1013:1021 arg1 consequences 982:993 the structural consequences 967:993 the structural consequences of known clinical mutations 967:1021 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 6 8 theme high 731:734 arg1 similarity 736:745 high similarity 731:745 high similarity to those of known T2 RNase structures from plants 731:795 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 2 9 theme clinical 253:260 arg1 symptoms 262:269 clinical symptoms 253:269 clinical symptoms (psychomotor impairments, spasticity and epilepsy) 253:320 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 4 10 theme tumor 526:530 arg1 properties 544:553 tumor suppressing properties 526:553 tumor suppressing properties of RNase T2 526:565 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 6 11 from plants 790:795 arg1 structures 774:783 known T2 RNase structures 759:783 known T2 RNase structures from plants 759:795 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 4 12 theme T2 564:565 arg1 properties 544:553 tumor suppressing properties 526:553 tumor suppressing properties of RNase T2 526:565 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 3 13 theme family 448:453 arg1 member 426:431 the only member 417:431 the only member of the Rh/T2/S family of acidic hydrolases in humans 417:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 3 13 theme family 448:453 arg1 T2 411:412 RNase T2 405:412 RNase T2 405:412 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 8 14 theme action 1161:1166 arg1 mode 1153:1156 its mode 1149:1156 its mode of action independent of its enzymatic activity 1149:1204 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 5 15 theme human 671:675 arg1 T2 683:684 human RNase T2 671:684 human RNase T2 671:684 We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. 22735700 2 16 dep symptoms 262:269 arg1 spasticity 297:306 spasticity 297:306 spasticity 297:306 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 16 dep symptoms 262:269 arg1 impairments 284:294 psychomotor impairments 272:294 psychomotor impairments 272:294 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 16 dep symptoms 262:269 arg1 epilepsy 312:319 epilepsy 312:319 epilepsy 312:319 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 7 17 theme known 998:1002 arg1 mutations 1013:1021 known clinical mutations 998:1021 known clinical mutations 998:1021 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 3 18 theme only 421:424 arg1 member 426:431 the only member 417:431 the only member of the Rh/T2/S family of acidic hydrolases in humans 417:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 3 18 theme only 421:424 arg1 T2 411:412 RNase T2 405:412 RNase T2 405:412 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 7 19 from features 896:903 arg1 presence 920:927 presence 920:927 presence of bivalent cations 920:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 5 20 theme RNase 677:681 arg1 T2 683:684 human RNase T2 671:684 human RNase T2 671:684 We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. 22735700 7 21 theme clinical 1004:1011 arg1 mutations 1013:1021 known clinical mutations 998:1021 known clinical mutations 998:1021 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 6 22 theme core 715:718 arg1 fold 720:723 The α+β core fold 707:723 The α+β core fold 707:723 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 2 23 theme matter 233:238 arg1 lesions 240:246 multifocal white matter lesions 216:246 multifocal white matter lesions 216:246 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 3 24 theme RNase 405:409 arg1 member 426:431 the only member 417:431 the only member of the Rh/T2/S family of acidic hydrolases in humans 417:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 3 24 theme RNase 405:409 arg1 T2 411:412 RNase T2 405:412 RNase T2 405:412 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 8 25 theme human 1096:1100 arg1 T2 1108:1109 human RNase T2 1096:1109 human RNase T2 1096:1109 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 6 26 theme distinct 849:856 arg1 differences 869:879 distinct structural differences 849:879 distinct structural differences 849:879 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 6 27 theme α+β 711:713 arg1 fold 720:723 The α+β core fold 707:723 The α+β core fold 707:723 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 2 28 theme psychomotor 272:282 arg1 spasticity 297:306 spasticity 297:306 spasticity 297:306 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 28 theme psychomotor 272:282 arg1 impairments 284:294 psychomotor impairments 272:294 psychomotor impairments 272:294 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 28 theme psychomotor 272:282 arg1 epilepsy 312:319 epilepsy 312:319 epilepsy 312:319 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 1 29 theme human 139:143 arg1 brain 145:149 the human brain 135:149 the human brain 135:149 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 8 30 theme enzymatic 1187:1195 arg1 activity 1197:1204 its enzymatic activity 1183:1204 its enzymatic activity 1183:1204 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 8 31 theme further 1059:1065 arg1 insight 1067:1073 further insight 1059:1073 further insight into the function of human RNase T2 1059:1109 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 1 32 theme brain 145:149 arg1 disease 124:130 white matter disease 111:130 white matter disease of the human brain 111:149 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 8 33 theme T2 1108:1109 arg1 function 1084:1091 the function 1080:1091 the function of human RNase T2 1080:1109 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 4 34 theme recent 490:495 arg1 years 497:501 recent years 490:501 recent years 490:501 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 8 35 theme activity 1197:1204 arg1 independent 1168:1178 independent 1168:1178 independent 1168:1178 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 3 36 from hydrolases 465:474 arg1 humans 479:484 humans 479:484 humans 479:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 2 37 theme deficiency 377:386 arg1 pathomechanism 350:363 the pathomechanism 346:363 the pathomechanism of RNase T2 deficiency 346:386 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 8 38 theme RNase 1102:1106 arg1 T2 1108:1109 human RNase T2 1096:1109 human RNase T2 1096:1109 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 4 39 theme new 504:506 arg1 properties 544:553 tumor suppressing properties 526:553 tumor suppressing properties of RNase T2 526:565 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 4 39 theme new 504:506 arg1 functions 508:516 new functions 504:516 new functions such as tumor suppressing properties of RNase T2 504:565 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 7 40 theme structural 971:980 arg1 consequences 982:993 the structural consequences 967:993 the structural consequences of known clinical mutations 967:1021 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 1 41 theme human 76:80 arg1 T2 88:89 human RNase T2 76:89 human RNase T2 76:89 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 2 42 theme T2 374:375 arg1 deficiency 377:386 RNase T2 deficiency 368:386 RNase T2 deficiency 368:386 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 0 43 theme only 30:33 arg1 T2 47:48 the only human RNase T2 26:48 the only human RNase T2 26:48 Structure and activity of the only human RNase T2. 22735700 3 44 theme Rh/T2/S 440:446 arg1 family 448:453 the Rh/T2/S family 436:453 the Rh/T2/S family of acidic hydrolases in humans 436:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 7 45 from T2 914:915 arg1 presence 920:927 presence 920:927 presence of bivalent cations 920:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 2 46 theme lobe 201:204 arg1 cysts 206:210 bilateral temporal lobe cysts 182:210 bilateral temporal lobe cysts 182:210 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 47 theme RNase 368:372 arg1 deficiency 377:386 RNase T2 deficiency 368:386 RNase T2 deficiency 368:386 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 4 48 theme catalytic 614:622 arg1 activity 624:631 its catalytic activity 610:631 its catalytic activity 610:631 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 5 49 theme X-ray 652:656 arg1 structure 658:666 the X-ray structure 648:666 the X-ray structure of human RNase T2 at 1.6 Å resolution 648:704 We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. 22735700 6 50 theme RNase 768:772 arg1 structures 774:783 known T2 RNase structures 759:783 known T2 RNase structures from plants 759:795 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 2 51 theme temporal 192:199 arg1 cysts 206:210 bilateral temporal lobe cysts 182:210 bilateral temporal lobe cysts 182:210 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 7 52 from presence 920:927 arg1 features 896:903 The catalytic features 882:903 The catalytic features of RNase T2 in presence of bivalent cations 882:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 0 53 theme RNase 41:45 arg1 T2 47:48 the only human RNase T2 26:48 the only human RNase T2 26:48 Structure and activity of the only human RNase T2. 22735700 7 54 theme cations 941:947 arg1 presence 920:927 presence 920:927 presence of bivalent cations 920:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 6 55 theme T2 765:766 arg1 structures 774:783 known T2 RNase structures 759:783 known T2 RNase structures from plants 759:795 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 2 56 theme bilateral 182:190 arg1 cysts 206:210 bilateral temporal lobe cysts 182:210 bilateral temporal lobe cysts 182:210 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 1 57 theme RNase 82:86 arg1 T2 88:89 human RNase T2 76:89 human RNase T2 76:89 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 0 58 theme human 35:39 arg1 T2 47:48 the only human RNase T2 26:48 the only human RNase T2 26:48 Structure and activity of the only human RNase T2. 22735700 6 59 theme loop 831:834 arg1 regions 836:842 the external loop regions 818:842 the external loop regions 818:842 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 6 60 theme known 759:763 arg1 structures 774:783 known T2 RNase structures 759:783 known T2 RNase structures from plants 759:795 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 5 61 from resolution 695:704 arg1 structure 658:666 the X-ray structure 648:666 the X-ray structure of human RNase T2 at 1.6 Å resolution 648:704 We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. 22735700 7 62 theme RNase 908:912 arg1 T2 914:915 RNase T2 908:915 RNase T2 in presence of bivalent cations 908:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 1 63 theme T2 88:89 arg1 gene 68:71 the gene 64:71 the gene of human RNase T2 64:89 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 6 64 theme external 822:829 arg1 regions 836:842 the external loop regions 818:842 the external loop regions 818:842 The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. 22735700 2 65 theme brain 161:165 arg1 abnormalities 167:179 brain abnormalities 161:179 brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) 161:247 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 66 theme white 227:231 arg1 lesions 240:246 multifocal white matter lesions 216:246 multifocal white matter lesions 216:246 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 7 67 theme T2 914:915 arg1 features 896:903 The catalytic features 882:903 The catalytic features of RNase T2 in presence of bivalent cations 882:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 7 68 theme bivalent 932:939 arg1 cations 941:947 bivalent cations 932:947 bivalent cations 932:947 The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. 22735700 2 69 theme multifocal 216:225 arg1 lesions 240:246 multifocal white matter lesions 216:246 multifocal white matter lesions 216:246 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 3 70 theme hydrolases 465:474 arg1 family 448:453 the Rh/T2/S family 436:453 the Rh/T2/S family of acidic hydrolases in humans 436:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 4 71 theme suppressing 532:542 arg1 properties 544:553 tumor suppressing properties 526:553 tumor suppressing properties of RNase T2 526:565 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 5 72 theme T2 683:684 arg1 structure 658:666 the X-ray structure 648:666 the X-ray structure of human RNase T2 at 1.6 Å resolution 648:704 We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. 22735700 8 73 theme independent 1168:1178 arg1 action 1161:1166 action 1161:1166 action independent of its enzymatic activity 1161:1204 Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity. 22735700 4 74 theme RNase 558:562 arg1 T2 564:565 RNase T2 558:565 RNase T2 558:565 In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. 22735700 3 75 theme acidic 458:463 arg1 hydrolases 465:474 acidic hydrolases 458:474 acidic hydrolases in humans 458:484 RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. 22735700 2 76 dep abnormalities 167:179 arg1 lesions 240:246 multifocal white matter lesions 216:246 multifocal white matter lesions 216:246 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 2 76 dep abnormalities 167:179 arg1 cysts 206:210 bilateral temporal lobe cysts 182:210 bilateral temporal lobe cysts 182:210 Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. 22735700 0 77 theme T2 47:48 arg1 activity 14:21 activity 14:21 activity 14:21 Structure and activity of the only human RNase T2. 22735700 0 77 theme T2 47:48 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and activity of the only human RNase T2. 22735700 1 78 theme white 111:115 arg1 disease 124:130 white matter disease 111:130 white matter disease of the human brain 111:149 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 22735700 1 79 from Mutations 51:59 arg1 gene 68:71 the gene 64:71 the gene of human RNase T2 64:89 Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. 2498325 2 0 theme human 113:117 arg1 E 140:140 human apolipoprotein (apo) E 113:140 human apolipoprotein (apo) E 113:140 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 4 1 theme sugar 364:368 arg1 analysis 370:377 amino sugar analysis 358:377 amino sugar analysis 358:377 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 6 2 theme glycosylation 872:884 arg1 sites 886:890 glycosylation sites 872:890 glycosylation sites 872:890 Site-specific mutants of apoE, designed to eliminate or alter glycosylation sites, were expressed in HeLa cells by acute transfection. 2498325 3 3 theme carbohydrate 230:241 arg1 site 254:257 The carbohydrate attachment site 226:257 The carbohydrate attachment site of plasma apoE 226:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 3 3 theme carbohydrate 230:241 arg1 apoE 269:272 plasma apoE 262:272 plasma apoE 262:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 10 4 theme hamster 1550:1556 arg1 cells 1570:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 7 5 theme attachment 1076:1085 arg1 Thr194 1041:1046 Thr194 1041:1046 Thr194 1041:1046 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 7 5 theme attachment 1076:1085 arg1 site 1055:1058 the site 1051:1058 the site of carbohydrate attachment in these cells 1051:1100 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 7 5 theme attachment 1076:1085 arg1 attachment 1076:1085 carbohydrate attachment 1063:1085 carbohydrate attachment 1063:1085 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 4 6 theme Sequence 336:343 arg1 analysis 345:352 Sequence analysis 336:352 Sequence analysis 336:352 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 8 7 theme Apolipoprotein 1176:1189 arg1 Gly196----Ser 1207:1219 Gly196----Ser 1207:1219 Gly196----Ser 1207:1219 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 8 7 theme Apolipoprotein 1176:1189 arg1 E 1191:1191 Apolipoprotein E 1176:1191 Apolipoprotein E(Thr194----Asn,Gly196----Ser) 1176:1220 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 3 8 theme plasma 262:267 arg1 apoE 269:272 plasma apoE 262:272 plasma apoE 262:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 0 9 theme apolipoprotein 23:36 arg1 E 38:38 human apolipoprotein E 17:38 human apolipoprotein E 17:38 Glycosylation of human apolipoprotein E. 2498325 5 10 theme apoE 662:665 arg1 vectors 686:692 human apoE plasmid expression vectors 656:692 human apoE plasmid expression vectors 656:692 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 10 11 theme CHO 1585:1587 arg1 cells 1594:1598 CHO ldlD cells 1585:1598 CHO ldlD cells 1585:1598 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 5 12 theme human 656:660 arg1 vectors 686:692 human apoE plasmid expression vectors 656:692 human apoE plasmid expression vectors 656:692 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 4 13 theme peptide 387:393 arg1 analysis 370:377 amino sugar analysis 358:377 amino sugar analysis 358:377 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 13 theme peptide 387:393 arg1 analysis 345:352 Sequence analysis 336:352 Sequence analysis 336:352 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 6 14 theme HeLa 911:914 arg1 cells 916:920 HeLa cells 911:920 HeLa cells 911:920 Site-specific mutants of apoE, designed to eliminate or alter glycosylation sites, were expressed in HeLa cells by acute transfection. 2498325 9 15 gly N-glycosylated 1435:1448 arg1 apoE 1426:1429 this apoE 1421:1429 this apoE 1421:1429 Studies with tunicamycin indicated that this apoE was N-glycosylated at Asn194. 2498325 9 15 gly N-glycosylated 1435:1448 arg2 Asn194 1453:1458 Asn194 1453:1458 Asn194 1453:1458 Studies with tunicamycin indicated that this apoE was N-glycosylated at Asn194. 2498325 5 16 theme plasmid 667:673 arg1 vectors 686:692 human apoE plasmid expression vectors 656:692 human apoE plasmid expression vectors 656:692 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 5 17 theme attachment 744:753 arg1 sites 755:759 potential carbohydrate attachment sites 721:759 potential carbohydrate attachment sites 721:759 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 3 18 theme single 293:298 arg1 peptide 308:314 a single tryptic peptide 291:314 a single tryptic peptide (residues 192-206) 291:333 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 3 18 theme single 293:298 arg1 residues 317:324 residues 192-206 317:332 residues 192-206 317:332 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 4 19 theme carbohydrate 464:475 arg1 moiety 477:482 the carbohydrate moiety 460:482 the carbohydrate moiety 460:482 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 8 20 from position 1280:1287 arg1 N-glycosylation 1261:1275 N-glycosylation 1261:1275 N-glycosylation at position 194 1261:1291 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 12 21 dep secreted 1738:1745 arg1 confirms 1811:1818 confirms 1811:1818 confirms that glycosylation is not essential for secretion of apoE 1811:1876 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 5 22 theme Mammalian 584:592 arg1 cells 594:598 Mammalian cells 584:598 Mammalian cells that normally do not express apoE 584:632 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 8 23 theme apparent 1321:1328 arg1 weight 1340:1345 a higher apparent molecular weight 1312:1345 a higher apparent molecular weight than native, O-glycosylated apoE 1312:1378 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 5 24 theme potential 721:729 arg1 sites 755:759 potential carbohydrate attachment sites 721:759 potential carbohydrate attachment sites 721:759 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 1 25 theme threonine 77:85 arg1 site 69:72 The carbohydrate attachment site 41:72 The carbohydrate attachment site 41:72 The carbohydrate attachment site is threonine 194. 2498325 1 25 theme threonine 77:85 arg1 194 87:89 threonine 194 77:89 threonine 194 77:89 The carbohydrate attachment site is threonine 194. 2498325 2 26 theme E 140:140 arg1 glycosylation 96:108 The glycosylation 92:108 The glycosylation of human apolipoprotein (apo) E 92:140 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 12 27 theme apoE 1873:1876 arg1 secretion 1860:1868 secretion 1860:1868 secretion of apoE 1860:1876 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 8 28 theme native 1352:1357 arg1 apoE 1375:1378 native, O-glycosylated apoE 1352:1378 native, O-glycosylated apoE 1352:1378 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 0 29 theme human 17:21 arg1 E 38:38 human apolipoprotein E 17:38 human apolipoprotein E 17:38 Glycosylation of human apolipoprotein E. 2498325 10 30 theme transfected 1468:1478 arg1 lines 1485:1489 Stably transfected cell lines 1461:1489 Stably transfected cell lines expressing human apoE 1461:1511 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 2 31 theme purified 160:167 arg1 apoE 176:179 purified plasma apoE 160:179 purified plasma apoE 160:179 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 12 32 gly glycosylation 1825:1837 arg1 apoE 1873:1876 apoE 1873:1876 apoE 1873:1876 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 0 33 theme E 38:38 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of human apolipoprotein E 0:38 Glycosylation of human apolipoprotein E. 2498325 9 34 from Asn194 1453:1458 arg1 N-glycosylated 1435:1448 N-glycosylated 1435:1448 N-glycosylated 1435:1448 Studies with tunicamycin indicated that this apoE was N-glycosylated at Asn194. 2498325 3 35 dep residues 317:324 arg1 192-206 326:332 192-206 326:332 192-206 326:332 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 12 36 theme glycosylation 1790:1802 arg1 absence 1779:1785 the absence 1775:1785 the absence of glycosylation 1775:1802 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 2 37 gly glycosylation 96:108 arg1 E 140:140 human apolipoprotein (apo) E 113:140 human apolipoprotein (apo) E 113:140 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 5 38 theme apoE 777:780 arg1 glycosylation 782:794 apoE glycosylation 777:794 apoE glycosylation 777:794 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 6 39 theme apoE 835:838 arg1 mutants 824:830 Site-specific mutants 810:830 Site-specific mutants 810:830 Site-specific mutants of apoE, designed to eliminate or alter glycosylation sites, were expressed in HeLa cells by acute transfection. 2498325 10 40 theme human 1502:1506 arg1 apoE 1508:1511 human apoE 1502:1511 human apoE 1502:1511 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 10 41 theme Chinese 1542:1548 arg1 cells 1570:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 1 42 theme carbohydrate 45:56 arg1 site 69:72 The carbohydrate attachment site 41:72 The carbohydrate attachment site 41:72 The carbohydrate attachment site is threonine 194. 2498325 1 42 theme carbohydrate 45:56 arg1 194 87:89 threonine 194 77:89 threonine 194 77:89 The carbohydrate attachment site is threonine 194. 2498325 2 43 theme apolipoprotein 119:132 arg1 E 140:140 human apolipoprotein (apo) E 113:140 human apolipoprotein (apo) E 113:140 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 12 44 theme high 1747:1750 arg1 levels 1752:1757 high levels 1747:1757 high levels of apoE 1747:1765 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 10 45 theme CHO 1565:1567 arg1 cells 1570:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 12 46 theme transfected 1710:1720 arg1 cells 1727:1731 The transfected ldlD cells 1706:1731 The transfected ldlD cells 1706:1731 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 10 47 theme ovary 1558:1562 arg1 cells 1570:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 11 48 theme transfected 1643:1653 arg1 cells 1665:1669 The transfected wild-type cells 1639:1669 The transfected wild-type cells 1639:1669 The transfected wild-type cells secreted multiply sialylated apoE. 2498325 7 49 theme carbohydrate 1063:1074 arg1 attachment 1076:1085 carbohydrate attachment 1063:1085 carbohydrate attachment 1063:1085 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 8 50 dep native 1352:1357 arg1 O-glycosylated 1360:1373 O-glycosylated 1360:1373 O-glycosylated 1360:1373 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 3 51 theme attachment 243:252 arg1 site 254:257 The carbohydrate attachment site 226:257 The carbohydrate attachment site of plasma apoE 226:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 3 51 theme attachment 243:252 arg1 apoE 269:272 plasma apoE 262:272 plasma apoE 262:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 12 52 theme ldlD 1722:1725 arg1 cells 1727:1731 The transfected ldlD cells 1706:1731 The transfected ldlD cells 1706:1731 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 2 53 theme apo 135:137 arg1 E 140:140 human apolipoprotein (apo) E 113:140 human apolipoprotein (apo) E 113:140 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 4 54 theme amino 358:362 arg1 analysis 370:377 amino sugar analysis 358:377 amino sugar analysis 358:377 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 5 55 theme expression 675:684 arg1 vectors 686:692 human apoE plasmid expression vectors 656:692 human apoE plasmid expression vectors 656:692 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 4 56 gly glycosylated 570:581 arg1 asialo-apoE 551:561 asialo-apoE 551:561 asialo-apoE 551:561 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 3 57 theme apoE 269:272 arg1 site 254:257 The carbohydrate attachment site 226:257 The carbohydrate attachment site of plasma apoE 226:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 3 57 theme apoE 269:272 arg1 apoE 269:272 plasma apoE 262:272 plasma apoE 262:272 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 6 58 theme acute 925:929 arg1 transfection 931:942 acute transfection 925:942 acute transfection 925:942 Site-specific mutants of apoE, designed to eliminate or alter glycosylation sites, were expressed in HeLa cells by acute transfection. 2498325 8 59 theme potential 1242:1250 arg1 site 1252:1255 a potential site 1240:1255 a potential site for N-glycosylation at position 194 1240:1291 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 10 60 theme ldlD 1589:1592 arg1 cells 1594:1598 CHO ldlD cells 1585:1598 CHO ldlD cells 1585:1598 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 5 61 from utilization 706:716 arg1 secretion 799:807 secretion 799:807 secretion 799:807 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 2 62 theme transfected 202:212 arg1 lines 219:223 transfected cell lines 202:223 transfected cell lines 202:223 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 6 63 gly glycosylation 872:884 arg2 sites 886:890 glycosylation sites 872:890 glycosylation sites 872:890 Site-specific mutants of apoE, designed to eliminate or alter glycosylation sites, were expressed in HeLa cells by acute transfection. 2498325 7 64 theme asialo 1009:1014 arg1 isoform 1016:1022 the asialo isoform 1005:1022 the asialo isoform 1005:1022 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 7 64 theme asialo 1009:1014 arg1 E 960:960 Apolipoprotein E 945:960 Apolipoprotein E(Thr194----Ala) 945:975 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 10 65 from defective 1611:1619 arg1 glycosylation 1624:1636 glycosylation 1624:1636 glycosylation 1624:1636 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 11 66 gly sialylated 1689:1698 arg1 apoE 1700:1703 multiply sialylated apoE 1680:1703 multiply sialylated apoE 1680:1703 The transfected wild-type cells secreted multiply sialylated apoE. 2498325 5 67 theme carbohydrate 731:742 arg1 sites 755:759 potential carbohydrate attachment sites 721:759 potential carbohydrate attachment sites 721:759 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 7 68 theme Apolipoprotein 945:958 arg1 Thr194----Ala 962:974 Thr194----Ala 962:974 Thr194----Ala 962:974 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 7 68 theme Apolipoprotein 945:958 arg1 isoform 1016:1022 the asialo isoform 1005:1022 the asialo isoform 1005:1022 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 7 68 theme Apolipoprotein 945:958 arg1 E 960:960 Apolipoprotein E 945:960 Apolipoprotein E(Thr194----Ala) 945:975 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 5 69 theme sites 755:759 arg1 utilization 706:716 the utilization 702:716 the utilization of potential carbohydrate attachment sites 702:759 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 5 69 theme sites 755:759 arg1 role 769:772 the role 765:772 the role of apoE glycosylation in secretion 765:807 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 8 70 gly N-glycosylation 1261:1275 arg1 position 1280:1287 position 194 1280:1291 position 194 1280:1291 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 8 70 gly N-glycosylation 1261:1275 arg2 position 1280:1287 position 194 1280:1291 position 194 1280:1291 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 3 71 theme tryptic 300:306 arg1 peptide 308:314 a single tryptic peptide 291:314 a single tryptic peptide (residues 192-206) 291:333 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 3 71 theme tryptic 300:306 arg1 residues 317:324 residues 192-206 317:332 residues 192-206 317:332 The carbohydrate attachment site of plasma apoE was localized to a single tryptic peptide (residues 192-206). 2498325 5 72 from role 769:772 arg1 secretion 799:807 secretion 799:807 secretion 799:807 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 1 73 theme attachment 58:67 arg1 site 69:72 The carbohydrate attachment site 41:72 The carbohydrate attachment site 41:72 The carbohydrate attachment site is threonine 194. 2498325 1 73 theme attachment 58:67 arg1 194 87:89 threonine 194 77:89 threonine 194 77:89 The carbohydrate attachment site is threonine 194. 2498325 8 74 theme higher 1314:1319 arg1 weight 1340:1345 a higher apparent molecular weight 1312:1345 a higher apparent molecular weight than native, O-glycosylated apoE 1312:1378 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 5 75 theme glycosylation 782:794 arg1 utilization 706:716 the utilization 702:716 the utilization of potential carbohydrate attachment sites 702:759 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 5 75 theme glycosylation 782:794 arg1 role 769:772 the role 765:772 the role of apoE glycosylation in secretion 765:807 Mammalian cells that normally do not express apoE were transfected with human apoE plasmid expression vectors to test the utilization of potential carbohydrate attachment sites and the role of apoE glycosylation in secretion. 2498325 7 76 from site 1055:1058 arg1 cells 1096:1100 these cells 1090:1100 these cells 1090:1100 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 8 77 theme molecular 1330:1338 arg1 weight 1340:1345 a higher apparent molecular weight 1312:1345 a higher apparent molecular weight than native, O-glycosylated apoE 1312:1378 Apolipoprotein E(Thr194----Asn,Gly196----Ser), which introduces a potential site for N-glycosylation at position 194, was secreted with a higher apparent molecular weight than native, O-glycosylated apoE. 2498325 7 78 theme apoE 1139:1142 arg1 glycosylation 1122:1134 glycosylation 1122:1134 glycosylation of apoE 1122:1142 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 12 79 theme apoE 1762:1765 arg1 levels 1752:1757 high levels 1747:1757 high levels of apoE 1747:1765 The transfected ldlD cells also secreted high levels of apoE even in the absence of glycosylation, which confirms that glycosylation is not essential for secretion of apoE. 2498325 10 80 theme cell 1480:1483 arg1 lines 1485:1489 Stably transfected cell lines 1461:1489 Stably transfected cell lines expressing human apoE 1461:1511 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 2 81 theme plasma 169:174 arg1 apoE 176:179 purified plasma apoE 160:179 purified plasma apoE 160:179 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 4 82 attach derived 395:401 arg2 peptide 387:393 this peptide 382:393 this peptide derived from asialo-, monosialo-, or disialo-apoE 382:443 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 82 attach derived 395:401 arg1 disialo-apoE 432:443 disialo-apoE 432:443 disialo-apoE 432:443 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 82 attach derived 395:401 arg1 asialo- 408:414 asialo- 408:414 asialo- 408:414 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 82 attach derived 395:401 arg1 monosialo- 417:426 monosialo- 417:426 monosialo- 417:426 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 0 83 gly Glycosylation 0:12 arg1 E 38:38 human apolipoprotein E 17:38 human apolipoprotein E 17:38 Glycosylation of human apolipoprotein E. 2498325 11 84 theme wild-type 1655:1663 arg1 cells 1665:1669 The transfected wild-type cells 1639:1669 The transfected wild-type cells 1639:1669 The transfected wild-type cells secreted multiply sialylated apoE. 2498325 7 85 gly glycosylation 1122:1134 arg1 apoE 1139:1142 apoE 1139:1142 apoE 1139:1142 Apolipoprotein E(Thr194----Ala) was secreted exclusively as the asialo isoform, confirming that Thr194 is the site of carbohydrate attachment in these cells and indicating that glycosylation of apoE is not essential for secretion. 2498325 9 86 with Studies 1381:1387 arg1 tunicamycin 1394:1404 tunicamycin 1394:1404 tunicamycin 1394:1404 Studies with tunicamycin indicated that this apoE was N-glycosylated at Asn194. 2498325 2 87 theme cell 214:217 arg1 lines 219:223 transfected cell lines 202:223 transfected cell lines 202:223 The glycosylation of human apolipoprotein (apo) E was examined with purified plasma apoE and apoE produced by transfected cell lines. 2498325 11 88 theme sialylated 1689:1698 arg1 apoE 1700:1703 multiply sialylated apoE 1680:1703 multiply sialylated apoE 1680:1703 The transfected wild-type cells secreted multiply sialylated apoE. 2498325 4 89 attach attached 487:494 arg1 Thr194 504:509 Thr194 504:509 Thr194 504:509 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 89 attach attached 487:494 arg2 moiety 477:482 the carbohydrate moiety 460:482 the carbohydrate moiety 460:482 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 89 attach attached 487:494 arg3 monosialo- 514:523 monosialo- 514:523 monosialo- 514:523 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 4 89 attach attached 487:494 arg3 disialo-apoE 529:540 disialo-apoE 529:540 disialo-apoE 529:540 Sequence analysis and amino sugar analysis of this peptide derived from asialo-, monosialo-, or disialo-apoE indicated that the carbohydrate moiety is attached only to Thr194 in monosialo- and disialo-apoE and that asialo-apoE is not glycosylated. 2498325 6 90 theme Site-specific 810:822 arg1 mutants 824:830 Site-specific mutants 810:830 Site-specific mutants 810:830 Site-specific mutants of apoE, designed to eliminate or alter glycosylation sites, were expressed in HeLa cells by acute transfection. 2498325 10 91 from glycosylation 1624:1636 arg1 defective 1611:1619 defective 1611:1619 defective 1611:1619 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 2498325 10 92 theme wild-type 1532:1540 arg1 cells 1570:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 wild-type Chinese hamster ovary (CHO) cells 1532:1574 Stably transfected cell lines expressing human apoE were prepared from wild-type Chinese hamster ovary (CHO) cells and from CHO ldlD cells, which are defective in glycosylation. 19737401 8 0 theme cysteine 1103:1110 arg1 residues 1112:1119 three cysteine residues 1097:1119 three cysteine residues present in the BST-2 extracellular domain 1097:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 8 1 from domain 1156:1161 arg1 present 1121:1127 present 1121:1127 present 1121:1127 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 9 2 theme wild 1357:1360 arg1 type 1362:1365 wild type 1357:1365 wild type 1357:1365 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 8 3 theme present 1121:1127 arg1 residues 1112:1119 three cysteine residues 1097:1119 three cysteine residues present in the BST-2 extracellular domain 1097:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 6 4 theme virus 877:881 arg1 release 883:889 HIV-1 virus release 871:889 HIV-1 virus release 871:889 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 3 5 theme cell 516:519 arg1 surface 521:527 the cell surface 512:527 the cell surface 512:527 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 19737401 1 6 theme infected 241:248 arg1 cells 250:254 infected cells 241:254 infected cells 241:254 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 12 7 theme BST-2 1946:1950 arg1 inhibition 1890:1899 inhibition 1890:1899 inhibition of virus release 1890:1916 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 7 theme BST-2 1946:1950 arg1 properties 1932:1941 separable properties 1922:1941 separable properties of BST-2 1922:1950 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 7 theme BST-2 1946:1950 arg1 sensitivity 1874:1884 Vpu sensitivity 1870:1884 Vpu sensitivity 1870:1884 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 1 8 theme Human 157:161 arg1 type 186:189 The Human Immunodeficiency virus type 1 153:191 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 1 8 theme Human 157:161 arg1 HIV-1 194:198 HIV-1 194:198 HIV-1 194:198 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 8 9 theme extracellular 1142:1154 arg1 domain 1156:1161 the BST-2 extracellular domain 1132:1161 the BST-2 extracellular domain 1132:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 5 10 link N-linked 738:745 arg1 glycosylation 747:759 N-linked glycosylation 738:759 N-linked glycosylation 738:759 We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. 19737401 1 11 theme virus 180:184 arg1 type 186:189 The Human Immunodeficiency virus type 1 153:191 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 1 11 theme virus 180:184 arg1 HIV-1 194:198 HIV-1 194:198 HIV-1 194:198 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 8 12 theme release 1226:1232 arg1 inhibition 1206:1215 inhibition 1206:1215 inhibition of virus release 1206:1232 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 8 12 theme release 1226:1232 arg1 sensitivity 1239:1249 sensitivity 1239:1249 sensitivity to Vpu 1239:1256 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 10 13 theme cell 1508:1511 arg1 surface 1513:1519 the cell surface 1504:1519 the cell surface of transfected 293T cells 1504:1545 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 4 14 theme BST-2 628:632 arg1 properties 614:623 structural properties 603:623 structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu 603:700 RESULTS: Here we defined structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu. 19737401 11 15 theme release 1757:1763 arg1 inhibition 1737:1746 inhibition 1737:1746 inhibition of virus release 1737:1763 CONCLUSION: While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. 19737401 5 16 theme extracellular 781:793 arg1 domain 795:800 the extracellular domain 777:800 the extracellular domain 777:800 We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. 19737401 6 17 theme surface 909:915 arg1 expression 917:926 surface expression 909:926 surface expression 909:926 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 10 18 theme 293T 1536:1539 arg1 cells 1541:1545 transfected 293T cells 1524:1545 transfected 293T cells 1524:1545 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 4 19 theme release 667:673 arg1 inhibition 647:656 inhibition 647:656 inhibition of virus release 647:673 RESULTS: Here we defined structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu. 19737401 12 20 theme separable 1922:1930 arg1 inhibition 1890:1899 inhibition 1890:1899 inhibition of virus release 1890:1916 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 20 theme separable 1922:1930 arg1 properties 1932:1941 separable properties 1922:1941 separable properties of BST-2 1922:1950 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 20 theme separable 1922:1930 arg1 sensitivity 1874:1884 Vpu sensitivity 1870:1884 Vpu sensitivity 1870:1884 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 5 21 theme N-linked 738:745 arg1 glycosylation 747:759 N-linked glycosylation 738:759 N-linked glycosylation 738:759 We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. 19737401 10 22 from defect 1437:1442 arg1 trafficking 1453:1463 BST-2 trafficking 1447:1463 BST-2 trafficking 1447:1463 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 11 23 theme cysteine-linked 1685:1699 arg1 dimers 1701:1706 cysteine-linked dimers 1685:1706 cysteine-linked dimers 1685:1706 CONCLUSION: While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. 19737401 0 24 theme HIV-1 89:93 arg1 release 101:107 HIV-1 virus release 89:107 HIV-1 virus release 89:107 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 10 25 theme similar 1577:1583 arg1 Vpu 1573:1575 Vpu 1573:1575 Vpu similar to wild type BST-2 1573:1602 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 0 26 theme release 101:107 arg1 inhibition 75:84 inhibition 75:84 inhibition of HIV-1 virus release 75:107 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 9 27 from cysteines 1296:1304 arg1 ectodomain 1313:1322 its ectodomain 1309:1322 its ectodomain 1309:1322 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 12 28 theme surface 1811:1817 arg1 expression 1819:1828 surface expression 1811:1828 surface expression 1811:1828 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 29 theme Vpu 1833:1835 arg1 sensitivity 1837:1847 Vpu sensitivity 1833:1847 Vpu sensitivity 1833:1847 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 10 30 theme gross 1431:1435 arg1 defect 1437:1442 a gross defect 1429:1442 a gross defect in BST-2 trafficking 1429:1463 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 3 31 theme current 415:421 arg1 model 431:435 A current working model 413:435 A current working model 413:435 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 19737401 10 32 theme wild 1588:1591 arg1 BST-2 1598:1602 wild type BST-2 1588:1602 wild type BST-2 1588:1602 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 0 33 theme dimers 33:38 arg1 important 61:69 important 61:69 important 61:69 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 0 33 theme dimers 33:38 arg1 formation 4:12 The formation 0:12 The formation of cysteine-linked dimers of BST-2/tetherin 0:56 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 8 34 theme triple 1048:1053 arg1 mutants 1064:1070 single, double, or triple cysteine mutants 1029:1070 single, double, or triple cysteine mutants 1029:1070 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 7 35 link cysteine-linked 993:1007 arg1 dimers 1009:1014 cysteine-linked dimers 993:1014 cysteine-linked dimers 993:1014 Rodent BST-2 was previously found to form cysteine-linked dimers. 19737401 0 36 theme BST-2/tetherin 43:56 arg1 dimers 33:38 cysteine-linked dimers 17:38 cysteine-linked dimers of BST-2/tetherin 17:56 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 1 37 theme CD4 295:297 arg1 degradation 280:290 proteasomal degradation 268:290 proteasomal degradation of CD4 268:297 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 8 38 theme mutants 1064:1070 arg1 Analysis 1017:1024 Analysis 1017:1024 Analysis of single, double, or triple cysteine mutants 1017:1070 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 6 39 theme N-linked 812:819 arg1 glycosylation 821:833 N-linked glycosylation 812:833 N-linked glycosylation 812:833 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 8 40 from present 1121:1127 arg1 domain 1156:1161 the BST-2 extracellular domain 1132:1161 the BST-2 extracellular domain 1132:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 3 41 theme tethering 483:491 arg1 particles 499:507 tethering viral particles 483:507 tethering viral particles 483:507 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 19737401 9 42 theme HIV-1 1384:1388 arg1 virions 1390:1396 Vpu-deficient HIV-1 virions 1370:1396 Vpu-deficient HIV-1 virions 1370:1396 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 12 43 theme virus 1904:1908 arg1 release 1910:1916 virus release 1904:1916 virus release 1904:1916 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 5 44 mod modified 726:733 arg3 glycosylation 747:759 N-linked glycosylation 738:759 N-linked glycosylation 738:759 We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. 19737401 5 44 mod modified 726:733 arg1 BST-2 717:721 BST-2 717:721 BST-2 717:721 We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. 19737401 6 45 link N-linked 812:819 arg1 glycosylation 821:833 N-linked glycosylation 812:833 N-linked glycosylation 812:833 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 1 46 theme virus 222:226 arg1 release 228:234 virus release 222:234 virus release from infected cells 222:254 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 6 47 theme release 883:889 arg1 inhibition 857:866 inhibition 857:866 inhibition of HIV-1 virus release 857:889 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 0 48 link cysteine-linked 17:31 arg1 dimers 33:38 cysteine-linked dimers 17:38 cysteine-linked dimers of BST-2/tetherin 17:56 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 9 49 theme type 1362:1365 arg1 release 1346:1352 release 1346:1352 release of wild type or Vpu-deficient HIV-1 virions 1346:1396 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 8 50 theme residues 1112:1119 arg1 one 1090:1092 one 1090:1092 one 1090:1092 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 8 50 theme residues 1112:1119 arg1 residues 1112:1119 three cysteine residues 1097:1119 three cysteine residues present in the BST-2 extracellular domain 1097:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 6 51 theme HIV-1 871:875 arg1 release 883:889 HIV-1 virus release 871:889 HIV-1 virus release 871:889 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 1 52 theme Immunodeficiency 163:178 arg1 type 186:189 The Human Immunodeficiency virus type 1 153:191 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 1 52 theme Immunodeficiency 163:178 arg1 HIV-1 194:198 HIV-1 194:198 HIV-1 194:198 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 9 53 theme Vpu-deficient 1370:1382 arg1 virions 1390:1396 Vpu-deficient HIV-1 virions 1370:1396 Vpu-deficient HIV-1 virions 1370:1396 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 10 54 theme transfected 1524:1534 arg1 cells 1541:1545 transfected 293T cells 1524:1545 transfected 293T cells 1524:1545 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 1 55 from cells 250:254 arg1 release 228:234 virus release 222:234 virus release from infected cells 222:254 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 9 56 theme virions 1390:1396 arg1 release 1346:1352 release 1346:1352 release of wild type or Vpu-deficient HIV-1 virions 1346:1396 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 8 57 theme BST-2 1136:1140 arg1 domain 1156:1161 the BST-2 extracellular domain 1132:1161 the BST-2 extracellular domain 1132:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 8 58 attach present 1121:1127 arg1 domain 1156:1161 the BST-2 extracellular domain 1132:1161 the BST-2 extracellular domain 1132:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 8 58 attach present 1121:1127 arg2 residues 1112:1119 three cysteine residues 1097:1119 three cysteine residues present in the BST-2 extracellular domain 1097:1161 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 1 59 theme type 186:189 arg1 protein 205:211 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 9 60 from contrast 1262:1269 arg1 unable 1328:1333 unable 1328:1333 unable 1328:1333 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 4 61 theme structural 603:612 arg1 properties 614:623 structural properties 603:623 structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu 603:700 RESULTS: Here we defined structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu. 19737401 12 62 theme Vpu 1870:1872 arg1 inhibition 1890:1899 inhibition 1890:1899 inhibition of virus release 1890:1916 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 62 theme Vpu 1870:1872 arg1 sensitivity 1874:1884 Vpu sensitivity 1870:1884 Vpu sensitivity 1870:1884 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 62 theme Vpu 1870:1872 arg1 properties 1932:1941 separable properties 1922:1941 separable properties of BST-2 1922:1950 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 10 63 theme cells 1541:1545 arg1 surface 1513:1519 the cell surface 1504:1519 the cell surface of transfected 293T cells 1504:1545 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 3 64 theme subsequent 554:563 arg1 endocytosis 565:575 their subsequent endocytosis 548:575 their subsequent endocytosis 548:575 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 19737401 12 65 theme dimerization 1782:1793 arg1 lack 1774:1777 lack 1774:1777 lack of dimerization 1774:1793 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 8 66 theme BST-2 1182:1186 arg1 dimerization 1188:1199 BST-2 dimerization 1182:1199 BST-2 dimerization 1182:1199 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 11 67 theme virus 1751:1755 arg1 release 1757:1763 virus release 1751:1763 virus release 1751:1763 CONCLUSION: While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. 19737401 7 68 theme cysteine-linked 993:1007 arg1 dimers 1009:1014 cysteine-linked dimers 993:1014 cysteine-linked dimers 993:1014 Rodent BST-2 was previously found to form cysteine-linked dimers. 19737401 12 69 theme release 1910:1916 arg1 inhibition 1890:1899 inhibition 1890:1899 inhibition of virus release 1890:1916 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 69 theme release 1910:1916 arg1 sensitivity 1874:1884 Vpu sensitivity 1870:1884 Vpu sensitivity 1870:1884 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 69 theme release 1910:1916 arg1 properties 1932:1941 separable properties 1922:1941 separable properties of BST-2 1922:1950 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 4 70 theme virus 661:665 arg1 release 667:673 virus release 661:673 virus release 661:673 RESULTS: Here we defined structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu. 19737401 5 71 from sites 768:772 arg1 domain 795:800 the extracellular domain 777:800 the extracellular domain 777:800 We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. 19737401 12 72 theme BST-2 1852:1856 arg1 expression 1819:1828 surface expression 1811:1828 surface expression 1811:1828 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 12 72 theme BST-2 1852:1856 arg1 sensitivity 1837:1847 Vpu sensitivity 1833:1847 Vpu sensitivity 1833:1847 However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2. 19737401 7 73 theme Rodent 951:956 arg1 BST-2 958:962 Rodent BST-2 951:962 Rodent BST-2 951:962 Rodent BST-2 was previously found to form cysteine-linked dimers. 19737401 11 74 link cysteine-linked 1685:1699 arg1 dimers 1701:1706 cysteine-linked dimers 1685:1706 cysteine-linked dimers 1685:1706 CONCLUSION: While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. 19737401 0 75 theme virus 95:99 arg1 release 101:107 HIV-1 virus release 89:107 HIV-1 virus release 89:107 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 11 76 theme dimers 1701:1706 arg1 formation 1672:1680 formation 1672:1680 formation of cysteine-linked dimers 1672:1706 CONCLUSION: While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. 19737401 8 77 theme single 1029:1034 arg1 mutants 1064:1070 single, double, or triple cysteine mutants 1029:1070 single, double, or triple cysteine mutants 1029:1070 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 8 78 theme virus 1220:1224 arg1 release 1226:1232 virus release 1220:1232 virus release 1220:1232 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 2 79 theme HIV-1 366:370 arg1 release 378:384 HIV-1 virus release 366:384 HIV-1 virus release 366:384 Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. 19737401 8 80 theme double 1037:1042 arg1 mutants 1064:1070 single, double, or triple cysteine mutants 1029:1070 single, double, or triple cysteine mutants 1029:1070 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 0 81 theme cysteine-linked 17:31 arg1 dimers 33:38 cysteine-linked dimers 17:38 cysteine-linked dimers of BST-2/tetherin 17:56 The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu. 19737401 2 82 theme sensitive 395:403 arg1 manner 405:410 a Vpu sensitive manner 389:410 a Vpu sensitive manner 389:410 Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. 19737401 6 83 gly glycosylation 821:833 arg1 surface 909:915 surface expression 909:926 surface expression 909:926 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 6 83 gly glycosylation 821:833 arg1 release 883:889 HIV-1 virus release 871:889 HIV-1 virus release 871:889 However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. 19737401 10 84 theme type 1593:1596 arg1 BST-2 1598:1602 wild type BST-2 1588:1602 wild type BST-2 1588:1602 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 1 85 theme proteasomal 268:278 arg1 degradation 280:290 proteasomal degradation 268:290 proteasomal degradation of CD4 268:297 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 8 86 theme cysteine 1055:1062 arg1 mutants 1064:1070 single, double, or triple cysteine mutants 1029:1070 single, double, or triple cysteine mutants 1029:1070 Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. 19737401 2 87 theme virus 372:376 arg1 release 378:384 HIV-1 virus release 366:384 HIV-1 virus release 366:384 Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. 19737401 3 88 theme working 423:429 arg1 model 431:435 A current working model 413:435 A current working model 413:435 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 19737401 2 89 theme host 340:343 arg1 BST-2/CD317 323:333 BST-2/CD317 323:333 BST-2/CD317 323:333 Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. 19737401 2 89 theme host 340:343 arg1 factor 345:350 a host factor 338:350 a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner 338:410 Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. 19737401 10 90 theme BST-2 1447:1451 arg1 trafficking 1453:1463 BST-2 trafficking 1447:1463 BST-2 trafficking 1447:1463 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 3 91 theme virus 466:470 arg1 release 472:478 virus release 466:478 virus release by tethering viral particles to the cell surface 466:527 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 19737401 9 92 from unable 1328:1333 arg1 contrast 1262:1269 contrast 1262:1269 contrast 1262:1269 In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. 19737401 11 93 theme BST-2 1623:1627 arg1 glycosylation 1629:1641 BST-2 glycosylation 1623:1641 BST-2 glycosylation 1623:1641 CONCLUSION: While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. 19737401 2 94 theme Recent 300:305 arg1 work 307:310 Recent work 300:310 Recent work 300:310 Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. 19737401 1 95 theme Vpu 201:203 arg1 protein 205:211 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein 153:211 BACKGROUND: The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. 19737401 10 96 theme mutant 1472:1477 arg1 protein 1479:1485 the mutant protein 1468:1485 the mutant protein 1468:1485 This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2. 19737401 3 97 theme viral 493:497 arg1 particles 499:507 tethering viral particles 483:507 tethering viral particles 483:507 A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis. 20823119 6 0 theme IgA1 878:881 arg1 protein 891:897 a Gal-deficient IgA1 myeloma protein 862:897 a Gal-deficient IgA1 myeloma protein 862:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 1 theme ion 1040:1042 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 1 theme ion 1040:1042 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 4 2 theme spectrometric 541:553 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 5 3 theme entire 630:635 arg1 range 637:641 the entire range 626:641 the entire range of O-glycan attachment sites 626:670 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 2 4 theme region 268:273 arg1 O-glycans 280:288 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 6 5 theme glycopeptides 845:857 arg1 glycoforms 824:833 all glycoforms 820:833 all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein 820:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 7 6 from localization 1199:1210 arg1 glycoforms 1266:1275 the six most abundant glycoforms 1244:1275 the six most abundant glycoforms 1244:1275 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 7 6 from localization 1199:1210 arg1 sites 1292:1296 the sites 1288:1296 the sites deficient in Gal 1288:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 6 7 with patients 931:938 arg1 IgAN 945:948 IgAN 945:948 IgAN 945:948 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 2 8 theme -deficient 251:260 arg1 O-glycans 280:288 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 1 9 theme primary 160:166 arg1 glomerulonephritis 168:185 the most common primary glomerulonephritis 144:185 the most common primary glomerulonephritis in the world 144:198 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 1 9 theme primary 160:166 arg1 nephropathy 122:132 IgA nephropathy 118:132 IgA nephropathy (IgAN) 118:139 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 9 10 theme kidney 1660:1665 arg1 disease 1667:1673 a chronic kidney disease 1650:1673 a chronic kidney disease 1650:1673 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 10 11 theme sites 1768:1772 arg1 analysis 1746:1753 the analysis 1742:1753 the analysis of clustered sites of O-glycosylation 1742:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 10 12 theme O-glycosylation 1777:1791 arg1 O-glycosylation 1777:1791 O-glycosylation 1777:1791 O-glycosylation 1777:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 10 12 theme O-glycosylation 1777:1791 arg1 sites 1768:1772 clustered sites 1758:1772 clustered sites of O-glycosylation 1758:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 6 13 theme aberrant 914:921 arg1 IgA1 923:926 the aberrant IgA1 910:926 the aberrant IgA1 in patients with IgAN 910:948 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 5 14 theme attachment 655:664 arg1 sites 666:670 O-glycan attachment sites 646:670 O-glycan attachment sites 646:670 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 2 15 theme glycosylated 212:223 arg1 IgA1 225:228 Aberrantly glycosylated IgA1 201:228 Aberrantly glycosylated IgA1 201:228 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 6 16 theme protein 891:897 arg1 glycopeptides 845:857 the HR glycopeptides 838:857 the HR glycopeptides of a Gal-deficient IgA1 myeloma protein 838:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 7 17 theme deficient 1298:1306 arg1 sites 1292:1296 the sites 1288:1296 the sites deficient in Gal 1288:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 1 18 from glomerulonephritis 168:185 arg1 world 194:198 the world 190:198 the world 190:198 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 8 19 theme LC-electron 1412:1422 arg1 analysis 1452:1459 on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis 1360:1459 analysis 1452:1459 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 7 20 theme fragments 1164:1172 arg1 variety 1145:1151 a variety 1143:1151 a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal 1143:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 7 20 theme fragments 1164:1172 arg1 fragments 1164:1172 IgA1 HR fragments 1156:1172 IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal 1156:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 8 21 theme dissociation 1433:1444 arg1 analysis 1452:1459 on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis 1360:1459 analysis 1452:1459 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 7 22 theme IgA1 1156:1159 arg1 fragments 1164:1172 IgA1 HR fragments 1156:1172 IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal 1156:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 0 23 theme electron 78:85 arg1 dissociation 104:115 electron capture/transfer dissociation 78:115 electron capture/transfer dissociation 78:115 Clustered O-glycans of IgA1: defining macro- and microheterogeneity by use of electron capture/transfer dissociation. 20823119 9 24 theme new 1491:1493 arg1 approach 1515:1522 a new clinically relevant approach 1489:1522 a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease 1489:1673 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 0 25 theme dissociation 104:115 arg1 use 71:73 use 71:73 use of electron capture/transfer dissociation 71:115 Clustered O-glycans of IgA1: defining macro- and microheterogeneity by use of electron capture/transfer dissociation. 20823119 7 26 from deficient 1298:1306 arg1 Gal 1311:1313 Gal 1311:1313 Gal 1311:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 6 27 theme AI-ECD 1015:1020 arg1 Fourier 1022:1028 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 5 28 theme threonine 733:741 arg1 residues 743:750 the clustered serine and threonine residues 708:750 the clustered serine and threonine residues in the HR of IgA1 heavy chain 708:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 2 29 theme disease 339:345 arg1 pathogenesis 319:330 the pathogenesis 315:330 the pathogenesis of the disease 315:345 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 7 30 theme IgA-specific 1111:1122 arg1 proteases 1124:1132 The IgA-specific proteases 1107:1132 The IgA-specific proteases 1107:1132 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 7 31 from Gal 1311:1313 arg1 deficient 1298:1306 deficient 1298:1306 deficient 1298:1306 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 6 32 theme trypsin 1003:1009 arg1 use 954:956 use 954:956 use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 954:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 4 33 theme dissociation 514:525 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 6 34 from IgA1 923:926 arg1 patients 931:938 patients 931:938 patients with IgAN 931:948 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 35 dep Fourier 1022:1028 arg1 transform 1030:1038 transform 1030:1038 transform 1030:1038 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 35 dep Fourier 1022:1028 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 35 dep Fourier 1022:1028 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 9 36 theme ECD/electron 1538:1549 arg1 fragmentation 1578:1590 ECD/electron transfer dissociation-type fragmentation 1538:1590 ECD/electron transfer dissociation-type fragmentation 1538:1590 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 6 37 theme proteases 991:999 arg1 combination 963:973 a combination 961:973 a combination of IgA-specific proteases 961:999 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 4 38 theme AI-ECD 528:533 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 9 39 theme dissociation-type 1560:1576 arg1 fragmentation 1578:1590 ECD/electron transfer dissociation-type fragmentation 1538:1590 ECD/electron transfer dissociation-type fragmentation 1538:1590 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 5 40 theme heavy 770:774 arg1 chain 776:780 IgA1 heavy chain 765:780 IgA1 heavy chain 765:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 7 41 gly O-glycosylation 1219:1233 arg2 sites 1235:1239 all O-glycosylation sites 1215:1239 all O-glycosylation sites 1215:1239 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 7 42 theme sites 1235:1239 arg1 localization 1199:1210 unambiguous localization 1187:1210 unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal 1187:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 4 43 theme ion-electron 493:504 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 8 44 theme on-line 1360:1366 arg1 LC 1391:1392 LC 1391:1392 LC 1391:1392 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 8 44 theme on-line 1360:1366 arg1 chromatography 1375:1388 on-line liquid chromatography 1360:1388 on-line liquid chromatography (LC) 1360:1393 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 3 45 theme specific 434:441 arg1 sites 443:447 specific sites 434:447 specific sites 434:447 It is not known whether the glycosylation defect occurs randomly or preferentially at specific sites. 20823119 4 46 theme IgA1 567:570 arg1 O-glycosylation 572:586 IgA1 O-glycosylation 567:586 IgA1 O-glycosylation 567:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 8 47 theme chromatography 1375:1388 arg1 MS/MS 1402:1406 on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis 1360:1459 MS/MS 1402:1406 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 10 48 theme applicable 1716:1725 arg1 principles 1727:1736 generally applicable principles 1706:1736 generally applicable principles 1706:1736 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 6 49 theme myeloma 883:889 arg1 protein 891:897 a Gal-deficient IgA1 myeloma protein 862:897 a Gal-deficient IgA1 myeloma protein 862:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 50 theme cyclotron 1044:1052 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 50 theme cyclotron 1044:1052 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 51 theme Gal-deficient 864:876 arg1 protein 891:897 a Gal-deficient IgA1 myeloma protein 862:897 a Gal-deficient IgA1 myeloma protein 862:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 4 52 theme mass 536:539 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 10 53 gly O-glycosylation 1777:1791 arg2 O-glycosylation 1777:1791 O-glycosylation 1777:1791 O-glycosylation 1777:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 10 53 gly O-glycosylation 1777:1791 arg2 sites 1768:1772 clustered sites 1758:1772 clustered sites of O-glycosylation 1758:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 4 54 theme analysis 555:562 arg1 utility 472:478 the utility 468:478 the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 468:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 6 55 theme tandem 1073:1078 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 55 theme tandem 1073:1078 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 2 56 theme hinge 262:266 arg1 region 268:273 hinge region 262:273 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 2 56 theme hinge 262:266 arg1 HR 276:277 HR 276:277 HR 276:277 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 1 57 theme common 153:158 arg1 glomerulonephritis 168:185 the most common primary glomerulonephritis 144:185 the most common primary glomerulonephritis in the world 144:198 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 1 57 theme common 153:158 arg1 nephropathy 122:132 IgA nephropathy 118:132 IgA nephropathy (IgAN) 118:139 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 6 58 theme HR 842:843 arg1 glycopeptides 845:857 the HR glycopeptides 838:857 the HR glycopeptides of a Gal-deficient IgA1 myeloma protein 838:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 9 59 theme chronic 1652:1658 arg1 disease 1667:1673 a chronic kidney disease 1650:1673 a chronic kidney disease 1650:1673 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 2 60 with IgA1 225:228 arg1 O-glycans 280:288 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 galactose (Gal)-deficient hinge region (HR) O-glycans 236:288 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 9 61 theme disease 1667:1673 arg1 pathogenesis 1634:1645 pathogenesis 1634:1645 pathogenesis of a chronic kidney disease 1634:1673 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 10 62 theme clustered 1758:1766 arg1 O-glycosylation 1777:1791 O-glycosylation 1777:1791 O-glycosylation 1777:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 10 62 theme clustered 1758:1766 arg1 sites 1768:1772 clustered sites 1758:1772 clustered sites of O-glycosylation 1758:1791 Furthermore, this work offers generally applicable principles for the analysis of clustered sites of O-glycosylation. 20823119 6 63 theme mass 1080:1083 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 63 theme mass 1080:1083 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 8 64 theme -AI-ECD 1394:1400 arg1 MS/MS 1402:1406 on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis 1360:1459 MS/MS 1402:1406 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 2 65 gly glycosylated 212:223 arg1 IgA1 225:228 Aberrantly glycosylated IgA1 201:228 Aberrantly glycosylated IgA1 201:228 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 5 66 theme O-glycan 646:653 arg1 sites 666:670 O-glycan attachment sites 646:670 O-glycan attachment sites 646:670 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 1 67 theme IgA 118:120 arg1 glomerulonephritis 168:185 the most common primary glomerulonephritis 144:185 the most common primary glomerulonephritis in the world 144:198 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 1 67 theme IgA 118:120 arg1 IgAN 135:138 IgAN 135:138 IgAN 135:138 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 1 67 theme IgA 118:120 arg1 nephropathy 122:132 IgA nephropathy 118:132 IgA nephropathy (IgAN) 118:139 IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. 20823119 6 68 gly glycopeptides 845:857 arg1 protein 891:897 a Gal-deficient IgA1 myeloma protein 862:897 a Gal-deficient IgA1 myeloma protein 862:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 68 gly glycopeptides 845:857 arg2 glycopeptides 845:857 the HR glycopeptides 838:857 the HR glycopeptides of a Gal-deficient IgA1 myeloma protein 838:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 5 69 theme sites 666:670 arg1 range 637:641 the entire range 626:641 the entire range of O-glycan attachment sites 626:670 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 6 70 theme combination 963:973 arg1 use 954:956 use 954:956 use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 954:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 8 71 theme transfer 1424:1431 arg1 analysis 1452:1459 on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis 1360:1459 analysis 1452:1459 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 9 72 dep new 1491:1493 arg1 relevant 1506:1513 relevant 1506:1513 relevant 1506:1513 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 7 73 theme HR 1161:1162 arg1 fragments 1164:1172 IgA1 HR fragments 1156:1172 IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal 1156:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 5 74 from residues 743:750 arg1 HR 759:760 the HR 755:760 the HR of IgA1 heavy chain 755:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 8 75 theme MS/MS 1446:1450 arg1 analysis 1452:1459 on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis 1360:1459 analysis 1452:1459 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 5 76 theme clustered 712:720 arg1 residues 743:750 the clustered serine and threonine residues 708:750 the clustered serine and threonine residues in the HR of IgA1 heavy chain 708:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 0 77 theme capture/transfer 87:102 arg1 dissociation 104:115 electron capture/transfer dissociation 78:115 electron capture/transfer dissociation 78:115 Clustered O-glycans of IgA1: defining macro- and microheterogeneity by use of electron capture/transfer dissociation. 20823119 5 78 theme serine 722:727 arg1 residues 743:750 the clustered serine and threonine residues 708:750 the clustered serine and threonine residues in the HR of IgA1 heavy chain 708:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 5 79 theme chain 776:780 arg1 HR 759:760 the HR 755:760 the HR of IgA1 heavy chain 755:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 7 80 theme abundant 1257:1264 arg1 glycoforms 1266:1275 the six most abundant glycoforms 1244:1275 the six most abundant glycoforms 1244:1275 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 7 80 theme abundant 1257:1264 arg1 sites 1292:1296 the sites 1288:1296 the sites deficient in Gal 1288:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 6 81 gly glycoforms 824:833 arg1 glycopeptides 845:857 the HR glycopeptides 838:857 the HR glycopeptides of a Gal-deficient IgA1 myeloma protein 838:897 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 3 82 theme glycosylation 376:388 arg1 defect 390:395 the glycosylation defect 372:395 the glycosylation defect 372:395 It is not known whether the glycosylation defect occurs randomly or preferentially at specific sites. 20823119 0 83 theme IgA1 23:26 arg1 O-glycans 10:18 O-glycans 10:18 O-glycans of IgA1 10:26 Clustered O-glycans of IgA1: defining macro- and microheterogeneity by use of electron capture/transfer dissociation. 20823119 6 84 theme resonance 1054:1062 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 84 theme resonance 1054:1062 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 9 85 theme transfer 1551:1558 arg1 fragmentation 1578:1590 ECD/electron transfer dissociation-type fragmentation 1538:1590 ECD/electron transfer dissociation-type fragmentation 1538:1590 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 6 86 theme IgA-specific 978:989 arg1 proteases 991:999 IgA-specific proteases 978:999 IgA-specific proteases 978:999 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 4 87 theme activated 483:491 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 5 88 theme IgA1 765:768 arg1 chain 776:780 IgA1 heavy chain 765:780 IgA1 heavy chain 765:780 However, locating and characterizing the entire range of O-glycan attachment sites are analytically challenging due to the clustered serine and threonine residues in the HR of IgA1 heavy chain. 20823119 7 89 theme O-glycosylation 1219:1233 arg1 sites 1235:1239 all O-glycosylation sites 1215:1239 all O-glycosylation sites 1215:1239 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 4 90 theme capture 506:512 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 2 91 theme pivotal 299:305 arg1 role 307:310 a pivotal role 297:310 a pivotal role 297:310 Aberrantly glycosylated IgA1, with galactose (Gal)-deficient hinge region (HR) O-glycans, plays a pivotal role in the pathogenesis of the disease. 20823119 6 92 theme FT-ICR 1065:1070 arg1 MS/MS 1099:1103 MS/MS 1099:1103 MS/MS 1099:1103 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 6 92 theme FT-ICR 1065:1070 arg1 spectrometry 1085:1096 ion cyclotron resonance (FT-ICR) tandem mass spectrometry 1040:1096 AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS) 1015:1104 To address this problem, we analyzed all glycoforms of the HR glycopeptides of a Gal-deficient IgA1 myeloma protein, mimicking the aberrant IgA1 in patients with IgAN, by use of a combination of IgA-specific proteases + trypsin and AI-ECD Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS). 20823119 4 93 theme O-glycosylation 572:586 arg1 analysis 555:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis 483:562 activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation 483:586 We have described the utility of activated ion-electron capture dissociation (AI-ECD) mass spectrometric analysis of IgA1 O-glycosylation. 20823119 9 94 theme molecular 1606:1614 arg1 events 1616:1621 the molecular events 1602:1621 the molecular events leading to pathogenesis of a chronic kidney disease 1602:1673 Our results thus represent a new clinically relevant approach that requires ECD/electron transfer dissociation-type fragmentation to define the molecular events leading to pathogenesis of a chronic kidney disease. 20823119 7 95 theme unambiguous 1187:1197 arg1 localization 1199:1210 unambiguous localization 1187:1210 unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal 1187:1313 The IgA-specific proteases provided a variety of IgA1 HR fragments that allowed unambiguous localization of all O-glycosylation sites in the six most abundant glycoforms, including the sites deficient in Gal. 20823119 8 96 theme liquid 1368:1373 arg1 LC 1391:1392 LC 1391:1392 LC 1391:1392 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 20823119 8 96 theme liquid 1368:1373 arg1 chromatography 1375:1388 on-line liquid chromatography 1360:1388 on-line liquid chromatography (LC) 1360:1393 Additionally, this protocol was adapted for on-line liquid chromatography (LC)-AI-ECD MS/MS and LC-electron transfer dissociation MS/MS analysis. 22588082 3 0 gly O-fucosylation 424:437 arg1 TSR 442:444 TSR 442:444 TSR 442:444 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 2 1 theme TSR 345:347 arg1 sequence 359:366 the TSR consensus sequence 341:366 the TSR consensus sequence for POFUT2-dependent fucosylation 341:400 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 7 2 theme substrate 1157:1165 arg1 specificity 1167:1177 substrate specificity 1157:1177 substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function 1157:1281 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 6 3 theme unusual 995:1001 arg1 structure 1006:1014 the unusual 3D structure 991:1014 the unusual 3D structure of a small part of the TSR 991:1041 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 0 4 theme repeat 63:68 arg1 fold 70:73 thrombospondin type 1 repeat fold 41:73 thrombospondin type 1 repeat fold 41:73 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 22588082 7 5 theme fold 1107:1110 arg1 motifs 1112:1117 distinct conserved 3D fold motifs 1085:1117 distinct conserved 3D fold motifs 1085:1117 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 1 6 theme serine/threonine 164:179 arg1 repeats 215:221 thrombospondin type 1 repeats 193:221 thrombospondin type 1 repeats (TSR) 193:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 6 theme serine/threonine 164:179 arg1 residues 181:188 serine/threonine residues 164:188 serine/threonine residues of thrombospondin type 1 repeats (TSR) 164:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 7 7 theme 3D 1104:1105 arg1 motifs 1112:1117 distinct conserved 3D fold motifs 1085:1117 distinct conserved 3D fold motifs 1085:1117 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 2 8 contain contain 333:339 arg1 proteins 324:331 >40 human proteins 314:331 >40 human proteins 314:331 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 2 8 contain contain 333:339 arg2 sequence 359:366 the TSR consensus sequence 341:366 the TSR consensus sequence for POFUT2-dependent fucosylation 341:400 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 7 9 theme distinct 1085:1092 arg1 motifs 1112:1117 distinct conserved 3D fold motifs 1085:1117 distinct conserved 3D fold motifs 1085:1117 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 6 10 theme artificial 868:877 arg1 substrate 888:896 an artificial mini-TSR substrate 865:896 an artificial mini-TSR substrate 865:896 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 7 11 theme sequence 1240:1247 arg1 diversity 1249:1257 very wide sequence diversity 1230:1257 very wide sequence diversity 1230:1257 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 1 12 located found 155:159 arg2 O-fucosylation 103:116 Protein O-fucosylation 95:116 Protein O-fucosylation 95:116 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 12 located found 155:159 arg1 residues 181:188 serine/threonine residues 164:188 serine/threonine residues of thrombospondin type 1 repeats (TSR) 164:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 12 located found 155:159 arg2 modification 142:153 a post-translational modification 121:153 a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR) 121:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 12 located found 155:159 arg1 repeats 215:221 thrombospondin type 1 repeats 193:221 thrombospondin type 1 repeats (TSR) 193:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 7 13 theme conserved 1094:1102 arg1 motifs 1112:1117 distinct conserved 3D fold motifs 1085:1117 distinct conserved 3D fold motifs 1085:1117 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 3 14 theme human 502:506 arg1 POFUT2 508:513 human POFUT2 502:513 human POFUT2 502:513 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 3 15 theme functional 479:488 arg1 analysis 490:497 a structural and functional analysis 462:497 a structural and functional analysis of human POFUT2 and its TSR substrate 462:535 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 5 16 theme wild-type 747:755 arg1 POFUT2 768:773 POFUT2 768:773 POFUT2 768:773 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 3 17 theme structural 464:473 arg1 analysis 490:497 a structural and functional analysis 462:497 a structural and functional analysis of human POFUT2 and its TSR substrate 462:535 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 2 18 theme protein 270:276 arg1 O-fucosyltransferase 278:297 protein O-fucosyltransferase 2 270:299 the protein O-fucosyltransferase 2 (POFUT2) 266:308 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 2 18 theme protein 270:276 arg1 POFUT2 302:307 POFUT2 302:307 POFUT2 302:307 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 2 19 theme POFUT2-dependent 372:387 arg1 fucosylation 389:400 POFUT2-dependent fucosylation 372:400 POFUT2-dependent fucosylation 372:400 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 4 20 theme acceptor 646:653 arg1 sites 663:667 TSR acceptor binding sites 642:667 TSR acceptor binding sites 642:667 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 7 21 theme motifs 1112:1117 arg1 recognition 1070:1080 recognition 1070:1080 recognition of distinct conserved 3D fold motifs 1070:1117 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 7 21 theme motifs 1112:1117 arg1 mechanism 1136:1144 a mechanism 1134:1144 a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function 1134:1281 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 3 22 theme substrate 527:535 arg1 analysis 490:497 a structural and functional analysis 462:497 a structural and functional analysis of human POFUT2 and its TSR substrate 462:535 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 4 23 theme TSR 642:644 arg1 sites 663:667 TSR acceptor binding sites 642:667 TSR acceptor binding sites 642:667 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 5 24 theme TSR 779:781 arg1 measurements 731:742 steady-state kinetic measurements 710:742 steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR 710:781 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 7 25 theme function 1274:1281 arg1 specificity 1167:1177 substrate specificity 1157:1177 substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function 1157:1281 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 7 26 theme diversity 1249:1257 arg1 specificity 1167:1177 substrate specificity 1157:1177 substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function 1157:1281 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 1 27 theme thrombospondin 193:206 arg1 repeats 215:221 thrombospondin type 1 repeats 193:221 thrombospondin type 1 repeats (TSR) 193:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 27 theme thrombospondin 193:206 arg1 TSR 224:226 TSR 224:226 TSR 224:226 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 5 28 theme catalytic 809:817 arg1 mechanism 819:827 the catalytic mechanism 805:827 the catalytic mechanism 805:827 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 6 29 theme TSR 956:958 arg1 sequence 968:975 the TSR protein sequence 952:975 the TSR protein sequence 952:975 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 1 30 theme type 208:211 arg1 repeats 215:221 thrombospondin type 1 repeats 193:221 thrombospondin type 1 repeats (TSR) 193:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 30 theme type 208:211 arg1 TSR 224:226 TSR 224:226 TSR 224:226 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 0 31 theme POFUT2 19:24 arg1 Structure 0:8 Structure 0:8 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 0:93 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 22588082 4 32 theme sugar 626:630 arg1 donor 632:636 sugar donor 626:636 sugar donor 626:636 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 0 33 theme human 13:17 arg1 POFUT2 19:24 human POFUT2 13:24 human POFUT2 13:24 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 22588082 6 34 theme protein 960:966 arg1 sequence 968:975 the TSR protein sequence 952:975 the TSR protein sequence 952:975 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 3 35 theme TSR 523:525 arg1 substrate 527:535 its TSR substrate 519:535 its TSR substrate 519:535 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 6 36 theme mini-TSR 879:886 arg1 substrate 888:896 an artificial mini-TSR substrate 865:896 an artificial mini-TSR substrate 865:896 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 1 37 theme Protein 95:101 arg1 O-fucosylation 103:116 Protein O-fucosylation 95:116 Protein O-fucosylation 95:116 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 37 theme Protein 95:101 arg1 modification 142:153 a post-translational modification 121:153 a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR) 121:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 38 theme repeats 215:221 arg1 repeats 215:221 thrombospondin type 1 repeats 193:221 thrombospondin type 1 repeats (TSR) 193:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 38 theme repeats 215:221 arg1 residues 181:188 serine/threonine residues 164:188 serine/threonine residues of thrombospondin type 1 repeats (TSR) 164:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 4 39 theme POFUT2 560:565 arg1 structures 546:555 Crystal structures 538:555 Crystal structures of POFUT2 538:565 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 5 40 theme steady-state 710:721 arg1 measurements 731:742 steady-state kinetic measurements 710:742 steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR 710:781 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 3 41 contain carried 450:456 arg1 we 447:448 we 447:448 we 447:448 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 3 41 contain carried 450:456 arg2 analysis 490:497 a structural and functional analysis 462:497 a structural and functional analysis of human POFUT2 and its TSR substrate 462:535 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 2 42 theme fucose 234:239 arg1 transfer 241:248 The fucose transfer 230:248 The fucose transfer 230:248 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 4 43 theme binding 655:661 arg1 sites 663:667 TSR acceptor binding sites 642:667 TSR acceptor binding sites 642:667 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 6 44 theme part 1027:1030 arg1 structure 1006:1014 the unusual 3D structure 991:1014 the unusual 3D structure of a small part of the TSR 991:1041 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 5 45 theme kinetic 723:729 arg1 measurements 731:742 steady-state kinetic measurements 710:742 steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR 710:781 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 6 46 theme 3D 1003:1004 arg1 structure 1006:1014 the unusual 3D structure 991:1014 the unusual 3D structure of a small part of the TSR 991:1041 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 6 47 theme small 1021:1025 arg1 part 1027:1030 a small part 1019:1030 a small part of the TSR 1019:1041 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 0 48 theme thrombospondin 41:54 arg1 fold 70:73 thrombospondin type 1 repeat fold 41:73 thrombospondin type 1 repeat fold 41:73 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 22588082 5 49 theme POFUT2 768:773 arg1 measurements 731:742 steady-state kinetic measurements 710:742 steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR 710:781 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 3 50 from O-fucosylation 424:437 arg1 TSR 442:444 TSR 442:444 TSR 442:444 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 4 51 theme Crystal 538:544 arg1 structures 546:555 Crystal structures 538:555 Crystal structures of POFUT2 538:565 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 5 52 theme Structural 670:679 arg1 findings 681:688 Structural findings 670:688 Structural findings 670:688 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 3 53 theme POFUT2 508:513 arg1 analysis 490:497 a structural and functional analysis 462:497 a structural and functional analysis of human POFUT2 and its TSR substrate 462:535 To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. 22588082 7 54 theme biological 1263:1272 arg1 function 1274:1281 biological function 1263:1281 biological function 1263:1281 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 7 55 theme wide 1235:1238 arg1 diversity 1249:1257 very wide sequence diversity 1230:1257 very wide sequence diversity 1230:1257 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 7 56 used used 1126:1129 arg2 mechanism 1136:1144 a mechanism 1134:1144 a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function 1134:1281 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 7 56 used used 1126:1129 arg2 recognition 1070:1080 recognition 1070:1080 recognition of distinct conserved 3D fold motifs 1070:1117 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 2 57 theme human 318:322 arg1 proteins 324:331 >40 human proteins 314:331 >40 human proteins 314:331 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 4 58 theme GT-B 603:606 arg1 fold 608:611 the classical GT-B fold 589:611 the classical GT-B fold 589:611 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 6 59 theme TSR 1039:1041 arg1 part 1027:1030 a small part 1019:1030 a small part of the TSR 1019:1041 By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. 22588082 1 60 theme post-translational 123:140 arg1 O-fucosylation 103:116 Protein O-fucosylation 95:116 Protein O-fucosylation 95:116 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 1 60 theme post-translational 123:140 arg1 modification 142:153 a post-translational modification 121:153 a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR) 121:227 Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). 22588082 4 61 theme classical 593:601 arg1 fold 608:611 the classical GT-B fold 589:611 the classical GT-B fold 589:611 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22588082 5 62 theme mutant 761:766 arg1 POFUT2 768:773 POFUT2 768:773 POFUT2 768:773 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 5 63 theme substrate 833:841 arg1 specificity 843:853 substrate specificity 833:853 substrate specificity 833:853 Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. 22588082 7 64 theme folded 1211:1216 arg1 proteins 1218:1225 completely folded proteins 1200:1225 completely folded proteins 1200:1225 Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function. 22588082 0 65 theme type 56:59 arg1 fold 70:73 thrombospondin type 1 repeat fold 41:73 thrombospondin type 1 repeat fold 41:73 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 22588082 0 66 dep Structure 0:8 arg1 insights 27:34 insights 27:34 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 0:93 Structure of human POFUT2: insights into thrombospondin type 1 repeat fold and O-fucosylation. 22588082 2 67 theme consensus 349:357 arg1 sequence 359:366 the TSR consensus sequence 341:366 the TSR consensus sequence for POFUT2-dependent fucosylation 341:400 The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. 22588082 4 68 theme fold 608:611 arg1 variation 576:584 a variation 574:584 a variation of the classical GT-B fold 574:611 Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. 22106953 0 0 theme atomic 82:87 arg1 basis 89:93 the atomic basis 78:93 the atomic basis for distinct roles in antigen processing 78:134 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 6 1 theme different 898:906 arg1 specificities 927:939 the different peptide N-terminal specificities 894:939 the different peptide N-terminal specificities between ERAP1 and ERAP2 894:963 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 2 2 theme first 362:366 arg1 structure 368:376 the first structure 358:376 the first structure of ERAP2 determined at 3.08 Å by X-ray crystallography 358:431 We report here the first structure of ERAP2 determined at 3.08 Å by X-ray crystallography. 22106953 5 3 theme large 741:745 arg1 cavity 756:761 A large internal cavity 739:761 A large internal cavity adjacent to the catalytic site 739:792 A large internal cavity adjacent to the catalytic site can accommodate large peptide substrates. 22106953 8 4 theme large 1241:1245 arg1 property 1275:1282 a key property 1269:1282 a key property of their biological role 1269:1307 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 4 theme large 1241:1245 arg1 sequences 1258:1266 sequences 1258:1266 sequences 1258:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 4 theme large 1241:1245 arg1 variety 1247:1253 a large variety 1239:1253 a large variety of sequences 1239:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 7 5 theme proposed 1112:1119 arg1 heterodimer 1133:1143 a proposed ERAP1-ERAP2 heterodimer 1110:1143 a proposed ERAP1-ERAP2 heterodimer 1110:1143 A noncrystallographic dimer observed may constitute a model for a proposed ERAP1-ERAP2 heterodimer. 22106953 1 6 theme mature 268:273 arg1 epitopes 275:282 mature epitopes 268:282 mature epitopes for binding to major histocompatibility class I molecules 268:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 7 7 theme ERAP1-ERAP2 1121:1131 arg1 heterodimer 1133:1143 a proposed ERAP1-ERAP2 heterodimer 1110:1143 a proposed ERAP1-ERAP2 heterodimer 1110:1143 A noncrystallographic dimer observed may constitute a model for a proposed ERAP1-ERAP2 heterodimer. 22106953 4 8 theme determined 681:690 arg1 structure 692:700 the recently determined structure 668:700 the recently determined structure of ERAP1 in its closed conformation 668:736 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 5 9 theme large 810:814 arg1 substrates 824:833 large peptide substrates 810:833 large peptide substrates 810:833 A large internal cavity adjacent to the catalytic site can accommodate large peptide substrates. 22106953 8 10 theme sequences 1258:1266 arg1 property 1275:1282 a key property 1269:1282 a key property of their biological role 1269:1307 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 10 theme sequences 1258:1266 arg1 sequences 1258:1266 sequences 1258:1266 sequences 1258:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 10 theme sequences 1258:1266 arg1 variety 1247:1253 a large variety 1239:1253 a large variety of sequences 1239:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 0 11 theme distinct 99:106 arg1 roles 108:112 distinct roles 99:112 distinct roles in antigen processing 99:134 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 5 12 theme internal 747:754 arg1 cavity 756:761 A large internal cavity 739:761 A large internal cavity adjacent to the catalytic site 739:792 A large internal cavity adjacent to the catalytic site can accommodate large peptide substrates. 22106953 8 13 theme role 1304:1307 arg1 property 1275:1282 a key property 1269:1282 a key property of their biological role 1269:1307 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 13 theme role 1304:1307 arg1 sequences 1258:1266 sequences 1258:1266 sequences 1258:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 13 theme role 1304:1307 arg1 variety 1247:1253 a large variety 1239:1253 a large variety of sequences 1239:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 3 14 theme active 518:523 arg1 site 525:528 the active site 514:528 the active site of the enzyme 514:542 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 3 14 theme active 518:523 arg1 enzyme 537:542 the enzyme 533:542 the enzyme 533:542 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 0 15 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human endoplasmic reticulum aminopeptidase 2 0:68 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 1 16 dep aminopeptidases 159:173 arg1 aminopeptidases 159:173 Endoplasmic reticulum aminopeptidases 137:173 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 137:189 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 16 dep aminopeptidases 159:173 arg1 ERAP2 185:189 ERAP2 185:189 ERAP2 185:189 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 16 dep aminopeptidases 159:173 arg1 ERAP1 175:179 ERAP1 175:179 ERAP1 175:179 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 6 17 theme structural 867:876 arg1 explanation 878:888 a structural explanation 865:888 a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 865:963 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 5 18 theme catalytic 779:787 arg1 site 789:792 the catalytic site 775:792 the catalytic site 775:792 A large internal cavity adjacent to the catalytic site can accommodate large peptide substrates. 22106953 4 19 theme ERAP1 705:709 arg1 structure 692:700 the recently determined structure 668:700 the recently determined structure of ERAP1 in its closed conformation 668:736 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 8 20 theme key 1271:1273 arg1 property 1275:1282 a key property 1269:1282 a key property of their biological role 1269:1307 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 20 theme key 1271:1273 arg1 sequences 1258:1266 sequences 1258:1266 sequences 1258:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 8 20 theme key 1271:1273 arg1 variety 1247:1253 a large variety 1239:1253 a large variety of sequences 1239:1266 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 2 21 theme ERAP2 381:385 arg1 structure 368:376 the first structure 358:376 the first structure of ERAP2 determined at 3.08 Å by X-ray crystallography 358:431 We report here the first structure of ERAP2 determined at 3.08 Å by X-ray crystallography. 22106953 5 22 theme peptide 816:822 arg1 substrates 824:833 large peptide substrates 810:833 large peptide substrates 810:833 A large internal cavity adjacent to the catalytic site can accommodate large peptide substrates. 22106953 0 23 theme antigen 117:123 arg1 processing 125:134 antigen processing 117:134 antigen processing 117:134 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 4 24 theme closed 718:723 arg1 conformation 725:736 its closed conformation 714:736 its closed conformation 714:736 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 3 25 theme electron 459:466 arg1 density 468:474 residual electron density 450:474 residual electron density 450:474 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 3 26 theme enzyme 537:542 arg1 site 525:528 the active site 514:528 the active site of the enzyme 514:542 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 3 26 theme enzyme 537:542 arg1 enzyme 537:542 the enzyme 533:542 the enzyme 533:542 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 2 27 theme X-ray 411:415 arg1 crystallography 417:431 X-ray crystallography 411:431 X-ray crystallography 411:431 We report here the first structure of ERAP2 determined at 3.08 Å by X-ray crystallography. 22106953 0 28 theme endoplasmic 31:41 arg1 aminopeptidase 53:66 human endoplasmic reticulum aminopeptidase 2 25:68 human endoplasmic reticulum aminopeptidase 2 25:68 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 1 29 theme major 299:303 arg1 molecules 332:340 major histocompatibility class I molecules 299:340 major histocompatibility class I molecules 299:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 4 30 theme domain 619:624 arg1 organization 626:637 The overall domain organization 607:637 The overall domain organization 607:637 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 4 30 theme domain 619:624 arg1 similar 649:655 similar 649:655 similar 649:655 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 1 31 theme vast 211:214 arg1 precursors 245:254 antigenic peptide precursors 227:254 antigenic peptide precursors 227:254 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 31 theme vast 211:214 arg1 variety 216:222 a vast variety 209:222 a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules 209:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 0 32 theme human 25:29 arg1 aminopeptidase 53:66 human endoplasmic reticulum aminopeptidase 2 25:68 human endoplasmic reticulum aminopeptidase 2 25:68 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 1 33 theme histocompatibility 305:322 arg1 molecules 332:340 major histocompatibility class I molecules 299:340 major histocompatibility class I molecules 299:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 3 34 theme residual 450:457 arg1 density 468:474 residual electron density 450:474 residual electron density 450:474 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 0 35 theme aminopeptidase 53:66 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human endoplasmic reticulum aminopeptidase 2 0:68 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 1 36 theme class 324:328 arg1 molecules 332:340 major histocompatibility class I molecules 299:340 major histocompatibility class I molecules 299:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 3 37 theme lysine 479:484 arg1 residue 486:492 a lysine residue 477:492 a lysine residue 477:492 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 0 38 theme reticulum 43:51 arg1 aminopeptidase 53:66 human endoplasmic reticulum aminopeptidase 2 25:68 human endoplasmic reticulum aminopeptidase 2 25:68 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 1 39 theme I 330:330 arg1 molecules 332:340 major histocompatibility class I molecules 299:340 major histocompatibility class I molecules 299:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 7 40 theme noncrystallographic 1048:1066 arg1 dimer 1068:1072 A noncrystallographic dimer 1046:1072 A noncrystallographic dimer observed 1046:1081 A noncrystallographic dimer observed may constitute a model for a proposed ERAP1-ERAP2 heterodimer. 22106953 0 41 from roles 108:112 arg1 processing 125:134 antigen processing 117:134 antigen processing 117:134 The crystal structure of human endoplasmic reticulum aminopeptidase 2 reveals the atomic basis for distinct roles in antigen processing. 22106953 6 42 theme such 983:986 arg1 differences 988:998 such differences 983:998 such differences 983:998 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 8 43 theme biological 1293:1302 arg1 role 1304:1307 their biological role 1287:1307 their biological role 1287:1307 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 6 44 theme N-terminal 916:925 arg1 specificities 927:939 the different peptide N-terminal specificities 894:939 the different peptide N-terminal specificities between ERAP1 and ERAP2 894:963 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 3 45 theme enzyme-product 583:596 arg1 complex 598:604 an enzyme-product complex 580:604 an enzyme-product complex 580:604 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 6 46 theme whole 1022:1026 arg1 sequence 1036:1043 the whole peptide sequence 1018:1043 the whole peptide sequence 1018:1043 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 4 47 from structure 692:700 arg1 conformation 725:736 its closed conformation 714:736 its closed conformation 714:736 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 6 48 theme peptide 1028:1034 arg1 sequence 1036:1043 the whole peptide sequence 1018:1043 the whole peptide sequence 1018:1043 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 8 49 theme homologous 1191:1200 arg1 aminopeptidases 1202:1216 two homologous aminopeptidases 1187:1216 two homologous aminopeptidases 1187:1216 Overall, the structure helps explain how two homologous aminopeptidases cooperate to process a large variety of sequences, a key property of their biological role. 22106953 1 50 theme antigenic 227:235 arg1 precursors 245:254 antigenic peptide precursors 227:254 antigenic peptide precursors 227:254 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 6 51 theme ERAP2 840:844 arg1 structure 846:854 The ERAP2 structure 836:854 The ERAP2 structure 836:854 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 4 52 theme overall 611:617 arg1 organization 626:637 The overall domain organization 607:637 The overall domain organization 607:637 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 4 52 theme overall 611:617 arg1 similar 649:655 similar 649:655 similar 649:655 The overall domain organization is highly similar to that of the recently determined structure of ERAP1 in its closed conformation. 22106953 1 53 theme Endoplasmic 137:147 arg1 reticulum 149:157 Endoplasmic reticulum 137:157 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 137:189 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 54 theme peptide 237:243 arg1 precursors 245:254 antigenic peptide precursors 227:254 antigenic peptide precursors 227:254 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 5 55 theme adjacent 763:770 arg1 cavity 756:761 A large internal cavity 739:761 A large internal cavity adjacent to the catalytic site 739:792 A large internal cavity adjacent to the catalytic site can accommodate large peptide substrates. 22106953 6 56 theme peptide 908:914 arg1 specificities 927:939 the different peptide N-terminal specificities 894:939 the different peptide N-terminal specificities between ERAP1 and ERAP2 894:963 The ERAP2 structure provides a structural explanation for the different peptide N-terminal specificities between ERAP1 and ERAP2 and suggests that such differences extend throughout the whole peptide sequence. 22106953 3 57 dep density 468:474 arg1 the 437:439 the 437:439 the 437:439 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 3 57 dep density 468:474 arg1 basis 441:445 basis 441:445 basis 441:445 On the basis of residual electron density, a lysine residue has been modeled in the active site of the enzyme; thus, the structure corresponds to an enzyme-product complex. 22106953 1 58 theme reticulum 149:157 arg1 aminopeptidases 159:173 Endoplasmic reticulum aminopeptidases 137:173 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 137:189 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 58 theme reticulum 149:157 arg1 ERAP2 185:189 ERAP2 185:189 ERAP2 185:189 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 58 theme reticulum 149:157 arg1 ERAP1 175:179 ERAP1 175:179 ERAP1 175:179 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 59 theme precursors 245:254 arg1 precursors 245:254 antigenic peptide precursors 227:254 antigenic peptide precursors 227:254 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 22106953 1 59 theme precursors 245:254 arg1 variety 216:222 a vast variety 209:222 a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules 209:340 Endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 cooperate to trim a vast variety of antigenic peptide precursors to generate mature epitopes for binding to major histocompatibility class I molecules. 16046623 6 0 gly glycosylated 1020:1031 arg1 protein 1040:1046 the protein 1036:1046 the protein used for crystallization 1036:1071 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 6 0 gly glycosylated 1020:1031 arg1 sites 1009:1013 the predicted sites 995:1013 the predicted sites 995:1013 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 6 0 gly glycosylated 1020:1031 arg2 all 988:990 all 988:990 all 988:990 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 6 0 gly glycosylated 1020:1031 arg1 all 988:990 all 988:990 all 988:990 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 6 0 gly glycosylated 1020:1031 arg2 sites 1009:1013 the predicted sites 995:1013 the predicted sites 995:1013 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 1 1 theme protein-1 159:167 arg1 expression 121:130 The expression 117:130 The expression of human vascular adhesion protein-1 (hVAP-1) 117:176 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 4 2 theme key 722:724 arg1 role 726:729 a key role 720:729 a key role 720:729 Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. 16046623 0 3 theme structural 67:76 arg1 features 78:85 unique structural features 60:85 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 0:115 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 0 4 with features 78:85 arg1 implications 103:114 functional implications 92:114 functional implications 92:114 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 3 5 theme intriguing 472:481 arg1 features 483:490 some intriguing features 467:490 some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling 467:667 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 5 6 theme sugar 861:865 arg1 units 867:871 sugar units 861:871 sugar units 861:871 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 0 7 theme functional 92:101 arg1 implications 103:114 functional implications 92:114 functional implications 92:114 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 3 8 theme fundamental 506:516 arg1 roles 518:522 fundamental roles 506:522 fundamental roles 506:522 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 4 9 theme substrate 693:701 arg1 channel 703:709 the substrate channel 689:709 the substrate channel 689:709 Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. 16046623 7 10 theme active 1152:1157 arg1 site 1159:1162 each active site 1147:1162 each active site 1147:1162 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 2 11 theme X-ray 312:316 arg1 structure 318:326 the X-ray structure 308:326 the X-ray structure 308:326 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 7 12 theme solvent-exposed 1102:1116 arg1 motif 1122:1126 a solvent-exposed RGD motif 1100:1126 a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 1100:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 7 13 from existence 1087:1095 arg1 entrance 1135:1142 the entrance 1131:1142 the entrance to each active site in hVAP-1 1131:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 6 14 theme predicted 999:1007 arg1 sites 1009:1013 the predicted sites 995:1013 the predicted sites 995:1013 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 7 15 theme RGD 1118:1120 arg1 motif 1122:1126 a solvent-exposed RGD motif 1100:1126 a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 1100:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 4 16 theme substrate 750:758 arg1 entry 760:764 the substrate entry 746:764 the substrate entry 746:764 Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. 16046623 7 17 theme motif 1122:1126 arg1 existence 1087:1095 the existence 1083:1095 the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 1083:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 2 18 theme other 403:407 arg1 CAOs 409:412 other CAOs 403:412 other CAOs 403:412 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 4 19 from channel 703:709 arg1 Leu469 679:684 Leu469 679:684 Leu469 at the substrate channel 679:709 Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. 16046623 3 20 theme adhesive 531:538 arg1 functions 554:562 the adhesive and enzymatic functions 527:562 the adhesive and enzymatic functions of hVAP-1 527:572 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 0 21 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 0:115 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 1 22 theme peripheral 267:276 arg1 tissue 278:283 the peripheral tissue 263:283 the peripheral tissue 263:283 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 2 23 theme amine 354:358 arg1 CAO 369:371 CAO 369:371 CAO 369:371 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 23 theme amine 354:358 arg1 hVAP-1 331:336 hVAP-1 331:336 hVAP-1 331:336 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 23 theme amine 354:358 arg1 oxidase 360:366 a human copper amine oxidase 339:366 a human copper amine oxidase (CAO) 339:372 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 24 theme human 341:345 arg1 CAO 369:371 CAO 369:371 CAO 369:371 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 24 theme human 341:345 arg1 hVAP-1 331:336 hVAP-1 331:336 hVAP-1 331:336 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 24 theme human 341:345 arg1 oxidase 360:366 a human copper amine oxidase 339:366 a human copper amine oxidase (CAO) 339:372 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 5 25 theme predicted 912:920 arg1 sites 938:942 the six predicted N-glycosylation sites 904:942 the six predicted N-glycosylation sites 904:942 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 7 26 theme functional 1202:1211 arg1 role 1213:1216 a functional role 1200:1216 a functional role 1200:1216 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 3 27 theme hVAP-1 608:613 arg1 role 600:603 the role 596:603 the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling 596:667 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 1 28 theme vascular 141:148 arg1 hVAP-1 170:175 hVAP-1 170:175 hVAP-1 170:175 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 28 theme vascular 141:148 arg1 protein-1 159:167 human vascular adhesion protein-1 135:167 human vascular adhesion protein-1 (hVAP-1) 135:176 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 0 29 theme vascular 31:38 arg1 protein-1 49:57 the human vascular adhesion protein-1 21:57 the human vascular adhesion protein-1 21:57 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 7 30 contain have 1195:1198 arg2 role 1213:1216 a functional role 1200:1216 a functional role 1200:1216 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 7 30 contain have 1195:1198 arg1 it 1188:1189 it 1188:1189 it 1188:1189 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 5 31 theme sites 938:942 arg1 two 897:899 two 897:899 two 897:899 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 5 31 theme sites 938:942 arg1 sites 938:942 the six predicted N-glycosylation sites 904:942 the six predicted N-glycosylation sites 904:942 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 0 32 theme human 25:29 arg1 protein-1 49:57 the human vascular adhesion protein-1 21:57 the human vascular adhesion protein-1 21:57 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 3 33 contain have 501:504 arg2 roles 518:522 fundamental roles 506:522 fundamental roles 506:522 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 3 33 contain have 501:504 arg1 features 483:490 some intriguing features 467:490 some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling 467:667 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 1 34 from blood 254:258 arg1 lymphocytes 237:247 lymphocytes 237:247 lymphocytes from blood 237:258 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 34 from blood 254:258 arg1 extravasation 220:232 extravasation 220:232 extravasation of lymphocytes from blood to the peripheral tissue 220:283 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 3 35 theme hVAP-1 567:572 arg1 functions 554:562 the adhesive and enzymatic functions 527:562 the adhesive and enzymatic functions of hVAP-1 527:572 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 1 36 theme inflammation 201:212 arg1 sites 192:196 sites 192:196 sites of inflammation 192:212 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 36 theme inflammation 201:212 arg1 inflammation 201:212 inflammation 201:212 inflammation 201:212 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 0 37 theme protein-1 49:57 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 0:115 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 3 38 from role 600:603 arg1 signaling 659:667 signaling 659:667 signaling 659:667 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 3 38 from role 600:603 arg1 inflammation 618:629 inflammation 618:629 inflammation 618:629 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 3 38 from role 600:603 arg1 attachment 643:652 lymphocyte attachment 632:652 lymphocyte attachment 632:652 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 5 39 located observed 885:892 arg1 two 897:899 two 897:899 two 897:899 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 5 39 located observed 885:892 arg1 sites 938:942 the six predicted N-glycosylation sites 904:942 the six predicted N-glycosylation sites 904:942 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 5 39 located observed 885:892 arg2 units 867:871 sugar units 861:871 sugar units 861:871 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 3 40 theme enzymatic 544:552 arg1 functions 554:562 the adhesive and enzymatic functions 527:562 the adhesive and enzymatic functions of hVAP-1 527:572 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 2 41 theme hVAP-1 331:336 arg1 structure 318:326 the X-ray structure 308:326 the X-ray structure 308:326 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 0 42 theme adhesion 40:47 arg1 protein-1 49:57 the human vascular adhesion protein-1 21:57 the human vascular adhesion protein-1 21:57 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 3 43 theme lymphocyte 632:641 arg1 attachment 643:652 lymphocyte attachment 632:652 lymphocyte attachment 632:652 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 4 44 theme active 838:843 arg1 site 845:848 the active site 834:848 the active site 834:848 Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. 16046623 7 45 from entrance 1135:1142 arg1 motif 1122:1126 a solvent-exposed RGD motif 1100:1126 a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 1100:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 7 45 from entrance 1135:1142 arg1 existence 1087:1095 the existence 1083:1095 the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 1083:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 7 45 from entrance 1135:1142 arg1 hVAP-1 1167:1172 hVAP-1 1167:1172 hVAP-1 1167:1172 Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. 16046623 1 46 from extravasation 220:232 arg1 blood 254:258 blood 254:258 blood 254:258 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 47 theme human 135:139 arg1 hVAP-1 170:175 hVAP-1 170:175 hVAP-1 170:175 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 47 theme human 135:139 arg1 protein-1 159:167 human vascular adhesion protein-1 135:167 human vascular adhesion protein-1 (hVAP-1) 135:176 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 0 48 theme unique 60:65 arg1 features 78:85 unique structural features 60:85 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 0:115 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 6 49 theme mutagenesis 955:965 arg1 analysis 967:974 mutagenesis analysis 955:974 mutagenesis analysis 955:974 Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. 16046623 0 50 dep structure 8:16 arg1 features 78:85 unique structural features 60:85 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 0:115 Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. 16046623 2 51 theme copper 347:352 arg1 CAO 369:371 CAO 369:371 CAO 369:371 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 51 theme copper 347:352 arg1 hVAP-1 331:336 hVAP-1 331:336 hVAP-1 331:336 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 2 51 theme copper 347:352 arg1 oxidase 360:366 a human copper amine oxidase 339:366 a human copper amine oxidase (CAO) 339:372 We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. 16046623 3 52 theme dimer 443:447 arg1 structure 449:457 The dimer structure 439:457 The dimer structure 439:457 The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. 16046623 5 53 gly N-glycosylation 922:936 arg2 sites 938:942 the six predicted N-glycosylation sites 904:942 the six predicted N-glycosylation sites 904:942 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 5 53 gly N-glycosylation 922:936 arg2 six 908:910 six 908:910 six 908:910 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 5 54 theme N-glycosylation 922:936 arg1 sites 938:942 the six predicted N-glycosylation sites 904:942 the six predicted N-glycosylation sites 904:942 Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. 16046623 1 55 theme adhesion 150:157 arg1 hVAP-1 170:175 hVAP-1 170:175 hVAP-1 170:175 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 55 theme adhesion 150:157 arg1 protein-1 159:167 human vascular adhesion protein-1 135:167 human vascular adhesion protein-1 (hVAP-1) 135:176 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 16046623 1 56 theme lymphocytes 237:247 arg1 extravasation 220:232 extravasation 220:232 extravasation of lymphocytes from blood to the peripheral tissue 220:283 The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. 17018531 11 0 theme fatty 1921:1925 arg1 acids 1927:1931 fatty acids 1921:1931 fatty acids 1921:1931 Binding was not detected for various glycolipids, phospholipids, or fatty acids. 17018531 1 1 theme other 247:251 arg1 lipids 253:258 other lipids 247:258 other lipids 247:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 4 2 theme purified 573:580 arg1 NPC2 582:585 Highly purified NPC2 566:585 Highly purified NPC2 566:585 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 3 3 theme functional 514:523 arg1 properties 525:534 the structural and functional properties 495:534 the structural and functional properties of recombinant human protein 495:563 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 9 4 theme chromatography-based 1431:1450 arg1 assays 1460:1465 chromatography-based binding assays 1431:1465 chromatography-based binding assays 1431:1465 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 5 5 theme endoglycosidase 874:888 arg1 H 890:890 endoglycosidase H 874:890 endoglycosidase H (Endo H) 874:899 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 5 theme endoglycosidase 874:888 arg1 H 898:898 Endo H 893:898 Endo H 893:898 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 3 6 theme structural 499:508 arg1 properties 525:534 the structural and functional properties 495:534 the structural and functional properties of recombinant human protein 495:563 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 12 7 theme sterol 2026:2031 arg1 transport 2033:2041 lysosomal sterol transport 2016:2041 lysosomal sterol transport 2016:2041 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 6 8 theme NPC2-deficient 1137:1150 arg1 fibroblasts 1152:1162 NPC2-deficient fibroblasts 1137:1162 NPC2-deficient fibroblasts 1137:1162 All glycoforms are endocytosed and ameliorate the cholesterol storage phenotype of NPC2-deficient fibroblasts. 17018531 4 9 gly glycosylated 620:631 arg1 isoforms 633:640 glycosylated isoforms 620:640 glycosylated isoforms 620:640 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 10 10 theme NPC2-deficient 1825:1838 arg1 liver 1846:1850 NPC2-deficient mouse liver 1825:1850 NPC2-deficient mouse liver 1825:1850 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 1 11 theme neurodegenerative 154:170 arg1 disorder 172:179 a fatal neurodegenerative disorder 146:179 a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids 146:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 1 11 theme neurodegenerative 154:170 arg1 disease 135:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 2 12 theme disease 277:283 arg1 form 265:268 One form 261:268 One form of the disease 261:283 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 4 13 theme glycosylated 620:631 arg1 isoforms 633:640 glycosylated isoforms 620:640 glycosylated isoforms 620:640 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 0 14 from disease 47:53 arg1 deficient 18:26 deficient 18:26 deficient 18:26 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 4 15 theme autopsy 683:689 arg1 specimens 691:699 human brain autopsy specimens 671:699 human brain autopsy specimens 671:699 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 10 16 theme cholesterol-related 1626:1644 arg1 molecules 1646:1654 cholesterol-related molecules 1626:1654 cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) 1626:1782 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 5 17 link N-linked 767:774 arg1 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 18 from protein 818:824 arg1 present 796:802 present 796:802 present 796:802 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 9 19 with complex 1511:1517 arg1 dehydroergosterol 1559:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 5 20 theme H-sensitive 1004:1014 arg1 oligosaccharides 1036:1051 Endo H-sensitive or Endo H-resistant oligosaccharides 999:1051 Endo H-sensitive or Endo H-resistant oligosaccharides 999:1051 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 21 theme Endo 893:896 arg1 H 890:890 endoglycosidase H 874:890 endoglycosidase H (Endo H) 874:899 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 21 theme Endo 893:896 arg1 H 898:898 Endo H 893:898 Endo H 893:898 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 9 22 theme cholesterol 1540:1550 arg1 dehydroergosterol 1559:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 4 23 theme human 671:675 arg1 specimens 691:699 human brain autopsy specimens 671:699 human brain autopsy specimens 671:699 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 0 24 theme multiple 68:75 arg1 glycoforms 77:86 multiple glycoforms 68:86 multiple glycoforms that bind a variety of sterols 68:117 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 4 25 located observed 659:666 arg2 that 654:657 that 654:657 that 654:657 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 4 25 located observed 659:666 arg1 specimens 691:699 human brain autopsy specimens 671:699 human brain autopsy specimens 671:699 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 5 26 from present 796:802 arg1 protein 818:824 the mature protein 807:824 the mature protein 807:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 0 27 from deficient 18:26 arg1 disease 47:53 Niemann-Pick C2 disease 31:53 Niemann-Pick C2 disease 31:53 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 2 28 gly glycoprotein 340:351 arg1 glycoprotein 340:351 a soluble lysosomal glycoprotein 320:351 a soluble lysosomal glycoprotein that binds cholesterol 320:374 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 2 28 gly glycoprotein 340:351 arg1 NPC2 314:317 NPC2 314:317 NPC2 314:317 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 8 29 theme cation-exchange 1359:1373 arg1 chromatography 1375:1388 cation-exchange chromatography 1359:1388 cation-exchange chromatography 1359:1388 All glycoforms bind cholesterol, and sterol binding to NPC2 significantly alters its behavior upon cation-exchange chromatography. 17018531 5 30 theme mature 811:816 arg1 protein 818:824 the mature protein 807:824 the mature protein 807:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 3 31 theme NPC2 425:428 arg1 function 413:420 the biological function 398:420 the biological function of NPC2 398:428 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 0 32 theme deficient 18:26 arg1 NPC2 0:3 NPC2 0:3 NPC2 0:3 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 0 32 theme deficient 18:26 arg1 protein 10:16 the protein 6:16 the protein deficient in Niemann-Pick C2 disease 6:53 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 5 33 theme N-linked 767:774 arg1 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 10 34 theme cholesterol 1734:1744 arg1 acetate 1746:1752 cholesterol acetate 1734:1752 cholesterol acetate 1734:1752 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 0 35 theme Niemann-Pick 31:42 arg1 disease 47:53 Niemann-Pick C2 disease 31:53 Niemann-Pick C2 disease 31:53 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 5 36 used utilized 952:959 arg2 Asn-116 932:938 Asn-116 932:938 Asn-116 932:938 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 1 37 theme endolysosomal 201:213 arg1 accumulation 215:226 an endolysosomal accumulation 198:226 an endolysosomal accumulation of cholesterol and other lipids 198:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 1 38 theme Niemann-Pick 120:131 arg1 disorder 172:179 a fatal neurodegenerative disorder 146:179 a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids 146:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 1 38 theme Niemann-Pick 120:131 arg1 disease 135:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 7 39 contain contains 1203:1210 arg2 mixture 1214:1220 a mixture 1212:1220 a mixture of both free and lipid-bound protein 1212:1257 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 7 39 contain contains 1203:1210 arg1 preparation 1191:1201 the purified preparation 1178:1201 the purified preparation 1178:1201 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 7 39 contain contains 1203:1210 arg2 preparation 1191:1201 the purified preparation 1178:1201 the purified preparation 1178:1201 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 7 39 contain contains 1203:1210 arg1 addition 1168:1175 addition 1168:1175 addition 1168:1175 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 2 40 theme lysosomal 330:338 arg1 glycoprotein 340:351 a soluble lysosomal glycoprotein 320:351 a soluble lysosomal glycoprotein that binds cholesterol 320:374 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 2 40 theme lysosomal 330:338 arg1 NPC2 314:317 NPC2 314:317 NPC2 314:317 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 0 41 theme C2 44:45 arg1 disease 47:53 Niemann-Pick C2 disease 31:53 Niemann-Pick C2 disease 31:53 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 7 42 theme lipid-bound 1239:1249 arg1 protein 1251:1257 both free and lipid-bound protein 1225:1257 both free and lipid-bound protein 1225:1257 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 10 43 dep molecules 1646:1654 arg1 precursors 1669:1678 cholesterol precursors 1657:1678 cholesterol precursors 1657:1678 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 10 43 dep molecules 1646:1654 arg1 5-alpha-cholestan-3-one 1759:1781 5-alpha-cholestan-3-one 1759:1781 5-alpha-cholestan-3-one 1759:1781 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 10 43 dep molecules 1646:1654 arg1 sterols 1687:1693 plant sterols 1681:1693 plant sterols 1681:1693 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 10 43 dep molecules 1646:1654 arg1 sulfate 1725:1731 cholesterol sulfate 1713:1731 cholesterol sulfate 1713:1731 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 10 43 dep molecules 1646:1654 arg1 oxysterols 1701:1710 some oxysterols 1696:1710 some oxysterols 1696:1710 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 10 43 dep molecules 1646:1654 arg1 acetate 1746:1752 cholesterol acetate 1734:1752 cholesterol acetate 1734:1752 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 2 44 from deficiency 300:309 arg1 glycoprotein 340:351 a soluble lysosomal glycoprotein 320:351 a soluble lysosomal glycoprotein that binds cholesterol 320:374 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 2 44 from deficiency 300:309 arg1 NPC2 314:317 NPC2 314:317 NPC2 314:317 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 7 45 theme free 1230:1233 arg1 protein 1251:1257 both free and lipid-bound protein 1225:1257 both free and lipid-bound protein 1225:1257 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 5 46 attach present 796:802 arg1 protein 818:824 the mature protein 807:824 the mature protein 807:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 46 attach present 796:802 arg2 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 1 47 theme lipids 253:258 arg1 accumulation 215:226 an endolysosomal accumulation 198:226 an endolysosomal accumulation of cholesterol and other lipids 198:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 3 48 theme recombinant 539:549 arg1 protein 557:563 recombinant human protein 539:563 recombinant human protein 539:563 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 5 49 attach linked 861:866 arg2 Asn-39 851:856 Asn-39 851:856 Asn-39 851:856 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 49 attach linked 861:866 arg1 oligosaccharide 911:925 an endoglycosidase H (Endo H)-sensitive oligosaccharide 871:925 an endoglycosidase H (Endo H)-sensitive oligosaccharide 871:925 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 9 50 theme analog 1552:1557 arg1 dehydroergosterol 1559:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 9 51 theme binding 1452:1458 arg1 assays 1460:1465 chromatography-based binding assays 1431:1465 chromatography-based binding assays 1431:1465 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 6 52 theme fibroblasts 1152:1162 arg1 phenotype 1124:1132 the cholesterol storage phenotype 1100:1132 the cholesterol storage phenotype of NPC2-deficient fibroblasts 1100:1162 All glycoforms are endocytosed and ameliorate the cholesterol storage phenotype of NPC2-deficient fibroblasts. 17018531 4 53 theme isoforms 633:640 arg1 mixture 609:615 a complex mixture 599:615 a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens 599:699 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 10 54 theme mouse 1840:1844 arg1 liver 1846:1850 NPC2-deficient mouse liver 1825:1850 NPC2-deficient mouse liver 1825:1850 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 1 55 theme fatal 148:152 arg1 disorder 172:179 a fatal neurodegenerative disorder 146:179 a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids 146:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 1 55 theme fatal 148:152 arg1 disease 135:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 12 56 theme lysosomal 2016:2024 arg1 transport 2033:2041 lysosomal sterol transport 2016:2041 lysosomal sterol transport 2016:2041 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 5 57 dep unmodified 975:984 arg1 either 962:967 either 962:967 either 962:967 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 58 used utilized 841:848 arg2 Asn-19 827:832 Asn-19 827:832 Asn-19 827:832 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 3 59 theme human 551:555 arg1 protein 557:563 recombinant human protein 539:563 recombinant human protein 539:563 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 6 60 theme storage 1116:1122 arg1 phenotype 1124:1132 the cholesterol storage phenotype 1100:1132 the cholesterol storage phenotype of NPC2-deficient fibroblasts 1100:1162 All glycoforms are endocytosed and ameliorate the cholesterol storage phenotype of NPC2-deficient fibroblasts. 17018531 4 61 theme complex 601:607 arg1 mixture 609:615 a complex mixture 599:615 a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens 599:699 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 10 62 theme cholesterol 1657:1667 arg1 precursors 1669:1678 cholesterol precursors 1657:1678 cholesterol precursors 1657:1678 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 0 63 theme sterols 111:117 arg1 sterols 111:117 sterols 111:117 sterols 111:117 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 0 63 theme sterols 111:117 arg1 variety 100:106 a variety 98:106 a variety of sterols 98:117 NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. 17018531 5 64 theme H-resistant 1024:1034 arg1 oligosaccharides 1036:1051 Endo H-sensitive or Endo H-resistant oligosaccharides 999:1051 Endo H-sensitive or Endo H-resistant oligosaccharides 999:1051 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 4 65 theme similar 643:649 arg1 isoforms 633:640 glycosylated isoforms 620:640 glycosylated isoforms 620:640 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 10 66 theme molecules 1646:1654 arg1 range 1617:1621 a range 1615:1621 a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) 1615:1782 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 7 67 theme purified 1182:1189 arg1 preparation 1191:1201 the purified preparation 1178:1201 the purified preparation 1178:1201 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 3 68 theme protein 557:563 arg1 properties 525:534 the structural and functional properties 495:534 the structural and functional properties of recombinant human protein 495:563 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 9 69 theme equimolar 1501:1509 arg1 complex 1511:1517 an equimolar complex 1498:1517 an equimolar complex with the fluorescent cholesterol analog dehydroergosterol 1498:1575 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 12 70 theme NPC2 2008:2011 arg1 function 1996:2003 a direct and specialized function 1971:2003 a direct and specialized function of NPC2 in lysosomal sterol transport 1971:2041 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 4 71 theme brain 677:681 arg1 specimens 691:699 human brain autopsy specimens 671:699 human brain autopsy specimens 671:699 Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. 17018531 9 72 theme fluorescent 1528:1538 arg1 dehydroergosterol 1559:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 the fluorescent cholesterol analog dehydroergosterol 1524:1575 Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. 17018531 8 73 theme sterol 1297:1302 arg1 binding 1304:1310 sterol binding 1297:1310 sterol binding to NPC2 1297:1318 All glycoforms bind cholesterol, and sterol binding to NPC2 significantly alters its behavior upon cation-exchange chromatography. 17018531 5 74 theme spectrometric 707:719 arg1 analysis 721:728 Mass spectrometric analysis 702:728 Mass spectrometric analysis 702:728 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 12 75 theme direct 1973:1978 arg1 function 1996:2003 a direct and specialized function 1971:2003 a direct and specialized function of NPC2 in lysosomal sterol transport 1971:2041 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 1 76 theme C 133:133 arg1 disorder 172:179 a fatal neurodegenerative disorder 146:179 a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids 146:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 1 76 theme C 133:133 arg1 disease 135:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease 120:141 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 17018531 5 77 theme Mass 702:705 arg1 analysis 721:728 Mass spectrometric analysis 702:728 Mass spectrometric analysis 702:728 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 12 78 theme specialized 1984:1994 arg1 function 1996:2003 a direct and specialized function 1971:2003 a direct and specialized function of NPC2 in lysosomal sterol transport 1971:2041 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 12 79 theme biochemical 1940:1950 arg1 properties 1952:1961 These biochemical properties 1934:1961 These biochemical properties 1934:1961 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 10 80 theme plant 1681:1685 arg1 sterols 1687:1693 plant sterols 1681:1693 plant sterols 1681:1693 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 5 81 theme present 796:802 arg1 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 3 82 dep understand 387:396 arg1 results 453:459 results 453:459 results in disease 453:470 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 3 83 theme biological 402:411 arg1 function 413:420 the biological function 398:420 the biological function of NPC2 398:428 To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. 17018531 5 84 theme -sensitive 900:909 arg1 oligosaccharide 911:925 an endoglycosidase H (Endo H)-sensitive oligosaccharide 871:925 an endoglycosidase H (Endo H)-sensitive oligosaccharide 871:925 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 85 theme potential 757:765 arg1 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 86 theme glycosylation 776:788 arg1 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 11 87 theme various 1882:1888 arg1 glycolipids 1890:1900 various glycolipids 1882:1900 various glycolipids 1882:1900 Binding was not detected for various glycolipids, phospholipids, or fatty acids. 17018531 6 88 theme cholesterol 1104:1114 arg1 phenotype 1124:1132 the cholesterol storage phenotype 1100:1132 the cholesterol storage phenotype of NPC2-deficient fibroblasts 1100:1162 All glycoforms are endocytosed and ameliorate the cholesterol storage phenotype of NPC2-deficient fibroblasts. 17018531 7 89 theme protein 1251:1257 arg1 mixture 1214:1220 a mixture 1212:1220 a mixture of both free and lipid-bound protein 1212:1257 In addition, the purified preparation contains a mixture of both free and lipid-bound protein. 17018531 10 90 theme cholesterol 1713:1723 arg1 sulfate 1725:1731 cholesterol sulfate 1713:1731 cholesterol sulfate 1713:1731 In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. 17018531 12 91 from function 1996:2003 arg1 transport 2033:2041 lysosomal sterol transport 2016:2041 lysosomal sterol transport 2016:2041 These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport. 17018531 5 92 gly glycosylation 776:788 arg2 three 751:755 three 751:755 three 751:755 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 5 92 gly glycosylation 776:788 arg2 sites 790:794 the three potential N-linked glycosylation sites 747:794 the three potential N-linked glycosylation sites present in the mature protein 747:824 Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. 17018531 2 93 theme soluble 322:328 arg1 glycoprotein 340:351 a soluble lysosomal glycoprotein 320:351 a soluble lysosomal glycoprotein that binds cholesterol 320:374 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 2 93 theme soluble 322:328 arg1 NPC2 314:317 NPC2 314:317 NPC2 314:317 One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. 17018531 1 94 theme cholesterol 231:241 arg1 accumulation 215:226 an endolysosomal accumulation 198:226 an endolysosomal accumulation of cholesterol and other lipids 198:258 Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. 11080501 8 0 theme cysteine-rich 1687:1699 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 0 theme cysteine-rich 1687:1699 arg1 motif 1733:1737 GA733 type 1 motif 1720:1737 GA733 type 1 motif 1720:1737 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 0 theme cysteine-rich 1687:1699 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 1 1 from level 290:294 arg1 epithelia 311:319 most normal epithelia 299:319 most normal epithelia 299:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 6 2 gly glycosylation 1275:1287 arg2 sites 1289:1293 glycosylation sites 1275:1293 glycosylation sites 1275:1293 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 8 3 theme thyroglobulin 1757:1769 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 3 theme thyroglobulin 1757:1769 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 3 theme thyroglobulin 1757:1769 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 1 4 theme most 237:240 arg1 carcinoma 265:273 most human gastrointestinal carcinoma 237:273 most human gastrointestinal carcinoma 237:273 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 3 5 theme disulfide-bonding 481:497 arg1 pattern 499:505 the disulfide-bonding pattern 477:505 the disulfide-bonding pattern of the GA733-2 antigen 477:528 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 8 6 theme 1A 1776:1777 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 6 theme 1A 1776:1777 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 6 theme 1A 1776:1777 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 6 7 link N-linked 1333:1340 arg1 carbohydrate 1342:1353 N-linked carbohydrate 1333:1353 N-linked carbohydrate 1333:1353 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 6 8 theme sites 1289:1293 arg1 Analysis 1263:1270 Analysis 1263:1270 Analysis of glycosylation sites 1263:1293 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 6 9 theme N-linked 1333:1340 arg1 carbohydrate 1342:1353 N-linked carbohydrate 1333:1353 N-linked carbohydrate 1333:1353 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 3 10 theme GA733-2 514:520 arg1 antigen 522:528 the GA733-2 antigen 510:528 the GA733-2 antigen 510:528 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 8 11 theme type 1793:1796 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 11 theme type 1793:1796 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 1 12 theme GA733-2 170:176 arg1 glycoprotein 204:215 a cell surface glycoprotein 189:215 a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia 189:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 1 12 theme GA733-2 170:176 arg1 antigen 178:184 The GA733-2 antigen 166:184 The GA733-2 antigen 166:184 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 7 13 theme glycosylation 1443:1455 arg1 sites 1457:1461 the three potential N-linked glycosylation sites 1414:1461 the three potential N-linked glycosylation sites 1414:1461 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 6 14 theme carbohydrate 1376:1387 arg1 groups 1389:1394 no O-linked carbohydrate groups 1364:1394 no O-linked carbohydrate groups 1364:1394 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 3 15 theme laser 567:571 arg1 spectrometry 600:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 1 16 theme gastrointestinal 248:263 arg1 carcinoma 265:273 most human gastrointestinal carcinoma 237:273 most human gastrointestinal carcinoma 237:273 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 7 17 theme potential 1424:1432 arg1 sites 1457:1461 the three potential N-linked glycosylation sites 1414:1461 the three potential N-linked glycosylation sites 1414:1461 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 4 18 theme growth 995:1000 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 5 19 theme disulfide 1153:1161 arg1 pattern 1163:1169 the recently determined disulfide pattern 1129:1169 the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1129:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 0 20 theme disulfide 17:25 arg1 assignments 32:42 disulfide bond assignments 17:42 disulfide bond assignments 17:42 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 8 21 theme nonglycosylated 1821:1835 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 21 theme nonglycosylated 1821:1835 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 21 theme nonglycosylated 1821:1835 arg1 motif 1876:1880 GA733 type 3 motif 1863:1880 GA733 type 3 motif 1863:1880 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 2 22 theme adhesion 442:449 arg1 molecules 451:459 adhesion molecules 442:459 adhesion molecules 442:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 0 23 theme human 77:81 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 1 24 theme normal 304:309 arg1 epithelia 311:319 most normal epithelia 299:319 most normal epithelia 299:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 0 25 theme carcinoma 100:108 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 1 26 gly glycoprotein 204:215 arg1 glycoprotein 204:215 a cell surface glycoprotein 189:215 a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia 189:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 1 26 gly glycoprotein 204:215 arg1 antigen 178:184 The GA733-2 antigen 166:184 The GA733-2 antigen 166:184 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 0 27 dep GA733-2 118:124 arg1 EGP 136:138 EGP 136:138 EGP 136:138 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 27 dep GA733-2 118:124 arg1 CO17-1A 127:133 CO17-1A 127:133 CO17-1A 127:133 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 27 dep GA733-2 118:124 arg1 KSA 148:150 KSA 148:150 KSA 148:150 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 27 dep GA733-2 118:124 arg1 Ep-CAM 157:162 Ep-CAM 157:162 Ep-CAM 157:162 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 27 dep GA733-2 118:124 arg1 KS1-4 141:145 KS1-4 141:145 KS1-4 141:145 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 4 28 theme expected 976:983 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 29 theme N 821:821 arg1 terminus 823:830 the N terminus 817:830 the N terminus 817:830 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 0 30 theme GA733-2 118:124 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 30 theme GA733-2 118:124 arg1 assignments 32:42 disulfide bond assignments 17:42 disulfide bond assignments 17:42 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 30 theme GA733-2 118:124 arg1 sites 64:68 N-glycosylation sites 48:68 N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 48:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 8 31 dep domains 1670:1676 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 31 dep domains 1670:1676 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 31 dep domains 1670:1676 arg1 motif 1733:1737 GA733 type 1 motif 1720:1737 GA733 type 1 motif 1720:1737 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 31 dep domains 1670:1676 arg1 motif 1876:1880 GA733 type 3 motif 1863:1880 GA733 type 3 motif 1863:1880 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 31 dep domains 1670:1676 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 31 dep domains 1670:1676 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 31 dep domains 1670:1676 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 32 theme type 1869:1872 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 32 theme type 1869:1872 arg1 motif 1876:1880 GA733 type 3 motif 1863:1880 GA733 type 3 motif 1863:1880 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 5 33 theme factor-binding 1230:1243 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 4 34 theme Numbering 771:779 arg1 GA733-2 781:787 Numbering GA733-2 771:787 Numbering GA733-2 771:787 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 2 35 theme cell-cell 339:347 arg1 It 322:323 It 322:323 It 322:323 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 2 35 theme cell-cell 339:347 arg1 protein 358:364 an unusual cell-cell adhesion protein 328:364 an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules 328:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 1 36 theme human 242:246 arg1 carcinoma 265:273 most human gastrointestinal carcinoma 237:273 most human gastrointestinal carcinoma 237:273 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 2 37 theme obvious 392:398 arg1 relationship 400:411 any obvious relationship 388:411 any obvious relationship to the four known classes of adhesion molecules 388:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 5 38 with consistent 1113:1122 arg1 pattern 1163:1169 the recently determined disulfide pattern 1129:1169 the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1129:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 4 39 dep Cys1-Cys4 872:880 arg1 pattern 926:932 a novel pattern 918:932 a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure 918:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 39 dep Cys1-Cys4 872:880 arg1 which 909:913 which 909:913 which 909:913 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 5 40 theme growth 1223:1228 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 3 41 theme purified 642:649 arg1 peptides 659:666 purified tryptic peptides 642:666 purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated 642:768 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 4 42 theme disulfide 849:857 arg1 Cys1-Cys4 872:880 Cys1-Cys4 872:880 Cys1-Cys4 872:880 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 42 theme disulfide 849:857 arg1 linkages 859:866 the first three disulfide linkages 833:866 the first three disulfide linkages 833:866 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 0 43 theme assignments 32:42 arg1 Determination 0:12 Determination 0:12 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 0:164 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 5 44 theme thyroglobulin 1178:1190 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 44 theme thyroglobulin 1178:1190 arg1 domain 1200:1205 the thyroglobulin type 1A domain 1174:1205 the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1174:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 4 45 theme structure 1024:1032 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 45 theme structure 1024:1032 arg1 domain 954:959 a cysteine-rich domain 938:959 a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure 938:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 5 46 theme disulfide 1050:1058 arg1 Cys7-Cys8 1073:1081 Cys7-Cys8 1073:1081 Cys7-Cys8 1073:1081 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 46 theme disulfide 1050:1058 arg1 linkages 1060:1067 The next three disulfide linkages 1035:1067 The next three disulfide linkages 1035:1067 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 8 47 theme antigen 1635:1641 arg1 domain 1613:1618 the extracellular domain 1595:1618 the extracellular domain of the GA733-2 antigen 1595:1641 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 47 theme antigen 1635:1641 arg1 antigen 1635:1641 the GA733-2 antigen 1623:1641 the GA733-2 antigen 1623:1641 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 2 48 theme molecules 451:459 arg1 classes 431:437 the four known classes 416:437 the four known classes of adhesion molecules 416:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 0 49 theme N-glycosylation 48:62 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 49 theme N-glycosylation 48:62 arg1 sites 64:68 N-glycosylation sites 48:68 N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 48:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 5 50 theme 1A 1197:1198 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 50 theme 1A 1197:1198 arg1 domain 1200:1205 the thyroglobulin type 1A domain 1174:1205 the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1174:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 3 51 theme mass 595:598 arg1 spectrometry 600:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 8 52 theme distinct 1661:1668 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 52 theme distinct 1661:1668 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 52 theme distinct 1661:1668 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 52 theme distinct 1661:1668 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 3 53 theme peptides 659:666 arg1 spectrometry 600:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 3 53 theme peptides 659:666 arg1 sequencing 628:637 N-terminal sequencing 617:637 N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated 617:768 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 4 54 theme first 837:841 arg1 Cys1-Cys4 872:880 Cys1-Cys4 872:880 Cys1-Cys4 872:880 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 54 theme first 837:841 arg1 linkages 859:866 the first three disulfide linkages 833:866 the first three disulfide linkages 833:866 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 8 55 theme type 1726:1729 arg1 motif 1733:1737 GA733 type 1 motif 1720:1737 GA733 type 1 motif 1720:1737 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 55 theme type 1726:1729 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 1 56 theme surface 196:202 arg1 glycoprotein 204:215 a cell surface glycoprotein 189:215 a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia 189:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 1 56 theme surface 196:202 arg1 antigen 178:184 The GA733-2 antigen 166:184 The GA733-2 antigen 166:184 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 5 57 theme next 1039:1042 arg1 Cys7-Cys8 1073:1081 Cys7-Cys8 1073:1081 Cys7-Cys8 1073:1081 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 57 theme next 1039:1042 arg1 linkages 1060:1067 The next three disulfide linkages 1035:1067 The next three disulfide linkages 1035:1067 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 4 58 theme disulfide 1014:1022 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 5 59 theme proteins 1245:1252 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 59 theme proteins 1245:1252 arg1 domain 1200:1205 the thyroglobulin type 1A domain 1174:1205 the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1174:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 6 60 link O-linked 1367:1374 arg1 groups 1389:1394 no O-linked carbohydrate groups 1364:1394 no O-linked carbohydrate groups 1364:1394 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 8 61 theme novel 1681:1685 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 61 theme novel 1681:1685 arg1 motif 1733:1737 GA733 type 1 motif 1720:1737 GA733 type 1 motif 1720:1737 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 61 theme novel 1681:1685 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 62 theme N-terminal 1701:1710 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 62 theme N-terminal 1701:1710 arg1 motif 1733:1737 GA733 type 1 motif 1720:1737 GA733 type 1 motif 1720:1737 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 62 theme N-terminal 1701:1710 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 6 63 theme GA733-2 1307:1313 arg1 antigen 1315:1321 GA733-2 antigen 1307:1321 GA733-2 antigen 1307:1321 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 8 64 theme cysteine-rich 1743:1755 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 64 theme cysteine-rich 1743:1755 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 64 theme cysteine-rich 1743:1755 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 7 65 gly glycosylated 1479:1490 arg1 Asn175 1464:1469 Asn175 1464:1469 Asn175 1464:1469 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 1 66 theme lower 284:288 arg1 level 290:294 a lower level 282:294 a lower level on most normal epithelia 282:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 3 67 theme antigen 522:528 arg1 pattern 499:505 the disulfide-bonding pattern 477:505 the disulfide-bonding pattern of the GA733-2 antigen 477:528 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 8 68 theme type 1771:1774 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 68 theme type 1771:1774 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 68 theme type 1771:1774 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 3 69 theme matrix-assisted 551:565 arg1 spectrometry 600:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 8 70 theme GA733 1787:1791 arg1 motif 1800:1804 GA733 type 2 motif 1787:1804 GA733 type 2 motif 1787:1804 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 70 theme GA733 1787:1791 arg1 domain 1779:1784 a cysteine-rich thyroglobulin type 1A domain 1741:1784 a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif) 1741:1805 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 7 71 gly glycosylation 1443:1455 arg2 sites 1457:1461 the three potential N-linked glycosylation sites 1414:1461 the three potential N-linked glycosylation sites 1414:1461 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 7 71 gly glycosylation 1443:1455 arg2 three 1418:1422 three 1418:1422 three 1418:1422 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 4 72 theme cysteine-rich 940:952 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 72 theme cysteine-rich 940:952 arg1 domain 954:959 a cysteine-rich domain 938:959 a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure 938:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 6 73 theme O-linked 1367:1374 arg1 groups 1389:1394 no O-linked carbohydrate groups 1364:1394 no O-linked carbohydrate groups 1364:1394 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 7 74 theme N-linked 1434:1441 arg1 sites 1457:1461 the three potential N-linked glycosylation sites 1414:1461 the three potential N-linked glycosylation sites 1414:1461 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 4 75 theme novel 920:924 arg1 pattern 926:932 a novel pattern 918:932 a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure 918:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 4 75 theme novel 920:924 arg1 which 909:913 which 909:913 which 909:913 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 5 76 dep proteins 1245:1252 arg1 6 1260:1260 6 1260:1260 6 1260:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 76 dep proteins 1245:1252 arg1 1 1254:1254 1 1254:1254 1 1254:1254 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 3 77 theme desorption/ionization 573:593 arg1 spectrometry 600:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 matrix-assisted laser desorption/ionization mass spectrometry 551:611 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 7 78 gly glycosylated 1560:1571 arg1 Asn51 1540:1544 Asn51 1540:1544 Asn51 1540:1544 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 4 79 theme epidermal 985:993 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 0 80 theme gastrointestinal 83:98 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 1 81 theme most 299:302 arg1 epithelia 311:319 most normal epithelia 299:319 most normal epithelia 299:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 8 82 theme unique 1814:1819 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 82 theme unique 1814:1819 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 82 theme unique 1814:1819 arg1 motif 1876:1880 GA733 type 3 motif 1863:1880 GA733 type 3 motif 1863:1880 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 4 83 theme factor-like 1002:1012 arg1 structure 1024:1032 the expected epidermal growth factor-like disulfide structure 972:1032 the expected epidermal growth factor-like disulfide structure 972:1032 Numbering GA733-2 cysteines sequentially from the N terminus, the first three disulfide linkages are Cys1-Cys4, Cys2-Cys6, and Cys3-Cys5, which is a novel pattern for a cysteine-rich domain instead of the expected epidermal growth factor-like disulfide structure. 11080501 0 84 theme antigen 110:116 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 6 85 contain contained 1323:1331 arg2 carbohydrate 1342:1353 N-linked carbohydrate 1333:1353 N-linked carbohydrate 1333:1353 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 6 85 contain contained 1323:1331 arg1 antigen 1315:1321 GA733-2 antigen 1307:1321 GA733-2 antigen 1307:1321 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 8 86 gly nonglycosylated 1821:1835 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 86 gly nonglycosylated 1821:1835 arg1 domains 1670:1676 three distinct domains 1655:1676 three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1655:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 86 gly nonglycosylated 1821:1835 arg1 motif 1876:1880 GA733 type 3 motif 1863:1880 GA733 type 3 motif 1863:1880 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 87 theme extracellular 1599:1611 arg1 domain 1613:1618 the extracellular domain 1595:1618 the extracellular domain of the GA733-2 antigen 1595:1641 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 87 theme extracellular 1599:1611 arg1 antigen 1635:1641 the GA733-2 antigen 1623:1641 the GA733-2 antigen 1623:1641 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 2 88 theme adhesion 349:356 arg1 It 322:323 It 322:323 It 322:323 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 2 88 theme adhesion 349:356 arg1 protein 358:364 an unusual cell-cell adhesion protein 328:364 an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules 328:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 2 89 theme known 425:429 arg1 classes 431:437 the four known classes 416:437 the four known classes of adhesion molecules 416:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 5 90 theme determined 1142:1151 arg1 pattern 1163:1169 the recently determined disulfide pattern 1129:1169 the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1129:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 8 91 theme GA733 1863:1867 arg1 domain 1837:1842 a unique nonglycosylated domain 1812:1842 a unique nonglycosylated domain without cysteines (GA733 type 3 motif) 1812:1881 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 91 theme GA733 1863:1867 arg1 motif 1876:1880 GA733 type 3 motif 1863:1880 GA733 type 3 motif 1863:1880 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 2 92 theme unusual 331:337 arg1 It 322:323 It 322:323 It 322:323 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 2 92 theme unusual 331:337 arg1 protein 358:364 an unusual cell-cell adhesion protein 328:364 an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules 328:459 It is an unusual cell-cell adhesion protein that does not exhibit any obvious relationship to the four known classes of adhesion molecules. 11080501 0 93 theme bond 27:30 arg1 assignments 32:42 disulfide bond assignments 17:42 disulfide bond assignments 17:42 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 5 94 theme insulin-like 1210:1221 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 6 95 theme glycosylation 1275:1287 arg1 sites 1289:1293 glycosylation sites 1275:1293 glycosylation sites 1275:1293 Analysis of glycosylation sites showed that GA733-2 antigen contained N-linked carbohydrate but that no O-linked carbohydrate groups were detected. 11080501 8 96 theme GA733-2 1627:1633 arg1 antigen 1635:1641 the GA733-2 antigen 1623:1641 the GA733-2 antigen 1623:1641 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 7 97 gly glycosylated 1522:1533 arg1 Asn88 1501:1505 Asn88 1501:1505 Asn88 1501:1505 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 7 98 link N-linked 1434:1441 arg1 sites 1457:1461 the three potential N-linked glycosylation sites 1414:1461 the three potential N-linked glycosylation sites 1414:1461 Of the three potential N-linked glycosylation sites, Asn175 was not glycosylated, whereas Asn88 was completely glycosylated, and Asn51 was partially glycosylated. 11080501 3 99 theme N-terminal 617:626 arg1 sequencing 628:637 N-terminal sequencing 617:637 N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated 617:768 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 0 100 theme sites 64:68 arg1 Determination 0:12 Determination 0:12 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 0:164 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 5 101 theme type 1192:1195 arg1 proteins 1245:1252 insulin-like growth factor-binding proteins 1 and 6 1210:1260 insulin-like growth factor-binding proteins 1 and 6 1210:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 5 101 theme type 1192:1195 arg1 domain 1200:1205 the thyroglobulin type 1A domain 1174:1205 the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1174:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 8 102 theme GA733 1720:1724 arg1 motif 1733:1737 GA733 type 1 motif 1720:1737 GA733 type 1 motif 1720:1737 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 8 102 theme GA733 1720:1724 arg1 domain 1712:1717 a novel cysteine-rich N-terminal domain 1679:1717 a novel cysteine-rich N-terminal domain (GA733 type 1 motif) 1679:1738 These data show that the extracellular domain of the GA733-2 antigen consists of three distinct domains; a novel cysteine-rich N-terminal domain (GA733 type 1 motif), a cysteine-rich thyroglobulin type 1A domain (GA733 type 2 motif), and a unique nonglycosylated domain without cysteines (GA733 type 3 motif). 11080501 5 103 theme domain 1200:1205 arg1 pattern 1163:1169 the recently determined disulfide pattern 1129:1169 the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6 1129:1260 The next three disulfide linkages are Cys7-Cys8, Cys9-Cys10, and Cys11-Cys12, consistent with the recently determined disulfide pattern of the thyroglobulin type 1A domain of insulin-like growth factor-binding proteins 1 and 6. 11080501 3 104 theme tryptic 651:657 arg1 peptides 659:666 purified tryptic peptides 642:666 purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated 642:768 In this study, the disulfide-bonding pattern of the GA733-2 antigen was determined using matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of purified tryptic peptides treated with 2-[2'-nitrophenylsulfonyl]-3-methyl-3-bromoindolenine or partially reduced and alkylated. 11080501 1 105 theme cell 191:194 arg1 glycoprotein 204:215 a cell surface glycoprotein 189:215 a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia 189:319 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 1 105 theme cell 191:194 arg1 antigen 178:184 The GA733-2 antigen 166:184 The GA733-2 antigen 166:184 The GA733-2 antigen is a cell surface glycoprotein highly expressed on most human gastrointestinal carcinoma and at a lower level on most normal epithelia. 11080501 0 106 gly N-glycosylation 48:62 arg2 sites 64:68 N-glycosylation sites 48:68 N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 48:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 106 gly N-glycosylation 48:62 arg2 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 11080501 0 106 gly N-glycosylation 48:62 arg1 GA733-2 118:124 the human gastrointestinal carcinoma antigen GA733-2 73:124 the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM) 73:163 Determination of disulfide bond assignments and N-glycosylation sites of the human gastrointestinal carcinoma antigen GA733-2 (CO17-1A, EGP, KS1-4, KSA, and Ep-CAM). 26146185 7 0 theme mass 853:856 arg1 analysis 871:878 Native mass spectrometry analysis 846:878 Native mass spectrometry analysis 846:878 Native mass spectrometry analysis showed that there is competition between the heterophilic and homophilic interactions. 26146185 2 1 theme receptor 324:331 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 1 theme receptor 324:331 arg1 domain 293:298 the N-terminal immunoglobulin superfamily (IgSF) domain 244:298 the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions 244:385 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 8 2 theme SRCR 1028:1031 arg1 domains 1033:1039 consecutive SRCR domains 1016:1039 consecutive SRCR domains 1016:1039 These data give insight into how interactions of consecutive SRCR domains are perturbed by SNPs and potential therapeutic reagents. 26146185 3 3 theme T-cell 488:493 arg1 activation 495:504 T-cell activation 488:504 T-cell activation 488:504 CD6 expression is mainly restricted to T cells, and the interaction between CD6 and CD166 regulates T-cell activation. 26146185 2 4 theme superfamily 274:284 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 4 theme superfamily 274:284 arg1 domain 293:298 the N-terminal immunoglobulin superfamily (IgSF) domain 244:298 the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions 244:385 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 4 5 theme SRCR 564:567 arg1 CD6 580:582 CD6 580:582 CD6 580:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 5 theme SRCR 564:567 arg1 domains 569:575 the three SRCR domains 554:575 the three SRCR domains of CD6 554:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 5 6 theme first 626:630 arg1 structure 632:640 This first structure 621:640 This first structure of consecutive SRCR domains 621:668 This first structure of consecutive SRCR domains reveals a nonlinear organization. 26146185 2 7 theme immunoglobulin 259:272 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 7 theme immunoglobulin 259:272 arg1 domain 293:298 the N-terminal immunoglobulin superfamily (IgSF) domain 244:298 the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions 244:385 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 1 8 theme transmembrane 85:97 arg1 CD6 76:78 CD6 76:78 CD6 76:78 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 1 8 theme transmembrane 85:97 arg1 protein 99:105 a transmembrane protein 83:105 a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains 83:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 7 9 theme heterophilic 925:936 arg1 interactions 953:964 the heterophilic and homophilic interactions 921:964 interactions 953:964 Native mass spectrometry analysis showed that there is competition between the heterophilic and homophilic interactions. 26146185 2 10 theme N-terminal 248:257 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 10 theme N-terminal 248:257 arg1 domain 293:298 the N-terminal immunoglobulin superfamily (IgSF) domain 244:298 the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions 244:385 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 6 11 from SNP 774:776 arg1 CD6 781:783 CD6 781:783 CD6 781:783 We characterized the binding sites on CD6 and CD166 and showed that a SNP in CD6 causes glycosylation that hinders the CD6/CD166 interaction. 26146185 1 12 theme scavenger 153:161 arg1 domains 193:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 8 13 theme domains 1033:1039 arg1 interactions 1000:1011 interactions 1000:1011 interactions of consecutive SRCR domains 1000:1039 These data give insight into how interactions of consecutive SRCR domains are perturbed by SNPs and potential therapeutic reagents. 26146185 1 14 theme receptor 163:170 arg1 domains 193:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 8 15 theme consecutive 1016:1026 arg1 domains 1033:1039 consecutive SRCR domains 1016:1039 consecutive SRCR domains 1016:1039 These data give insight into how interactions of consecutive SRCR domains are perturbed by SNPs and potential therapeutic reagents. 26146185 1 16 contain containing 136:145 arg2 domains 193:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 1 16 contain containing 136:145 arg1 region 129:134 an extracellular region 112:134 an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains 112:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 1 17 theme cysteine 172:179 arg1 domains 193:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 4 18 theme X-ray 526:530 arg1 structures 540:549 the X-ray crystal structures 522:549 the X-ray crystal structures of the three SRCR domains of CD6 522:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 1 19 with protein 99:105 arg1 region 129:134 an extracellular region 112:134 an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains 112:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 1 20 theme rich 181:184 arg1 domains 193:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 5 21 theme domains 662:668 arg1 structure 632:640 This first structure 621:640 This first structure of consecutive SRCR domains 621:668 This first structure of consecutive SRCR domains reveals a nonlinear organization. 26146185 2 22 theme IgSF 287:290 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 22 theme IgSF 287:290 arg1 domain 293:298 the N-terminal immunoglobulin superfamily (IgSF) domain 244:298 the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions 244:385 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 4 23 theme N-terminal 592:601 arg1 domains 603:609 two N-terminal domains 588:609 two N-terminal domains of CD166 588:618 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 23 theme N-terminal 592:601 arg1 domains 569:575 the three SRCR domains 554:575 the three SRCR domains of CD6 554:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 23 theme N-terminal 592:601 arg1 CD166 614:618 CD166 614:618 CD166 614:618 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 0 24 theme CD6 14:16 arg1 Structures 0:9 Structures 0:9 Structures of CD6 0:16 Structures of CD6 and Its Ligand CD166 Give Insight into Their Interaction. 26146185 0 24 theme CD6 14:16 arg1 CD166 33:37 Its Ligand CD166 22:37 Its Ligand CD166 22:37 Structures of CD6 and Its Ligand CD166 Give Insight into Their Interaction. 26146185 6 25 theme CD6/CD166 823:831 arg1 interaction 833:843 the CD6/CD166 interaction 819:843 the CD6/CD166 interaction 819:843 We characterized the binding sites on CD6 and CD166 and showed that a SNP in CD6 causes glycosylation that hinders the CD6/CD166 interaction. 26146185 1 26 theme SRCR 187:190 arg1 domains 193:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 three scavenger receptor cysteine rich (SRCR) domains 147:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 0 27 theme Ligand 26:31 arg1 CD166 33:37 Its Ligand CD166 22:37 Its Ligand CD166 22:37 Structures of CD6 and Its Ligand CD166 Give Insight into Their Interaction. 26146185 4 28 theme crystal 532:538 arg1 structures 540:549 the X-ray crystal structures 522:549 the X-ray crystal structures of the three SRCR domains of CD6 522:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 2 29 theme proximal 215:222 arg1 CD6 234:236 CD6 234:236 CD6 234:236 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 29 theme proximal 215:222 arg1 domain 224:229 The membrane proximal domain 202:229 The membrane proximal domain of CD6 202:236 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 3 30 theme CD6 388:390 arg1 expression 392:401 CD6 expression 388:401 CD6 expression 388:401 CD6 expression is mainly restricted to T cells, and the interaction between CD6 and CD166 regulates T-cell activation. 26146185 5 31 theme nonlinear 680:688 arg1 organization 690:701 a nonlinear organization 678:701 a nonlinear organization 678:701 This first structure of consecutive SRCR domains reveals a nonlinear organization. 26146185 2 32 theme membrane 206:213 arg1 CD6 234:236 CD6 234:236 CD6 234:236 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 32 theme membrane 206:213 arg1 domain 224:229 The membrane proximal domain 202:229 The membrane proximal domain of CD6 202:236 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 33 theme homophilic 363:372 arg1 interactions 374:385 homophilic interactions 363:385 homophilic interactions 363:385 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 34 theme surface 316:322 arg1 CD166 334:338 CD166 334:338 CD166 334:338 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 34 theme surface 316:322 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 8 35 theme potential 1067:1075 arg1 reagents 1089:1096 potential therapeutic reagents 1067:1096 potential therapeutic reagents 1067:1096 These data give insight into how interactions of consecutive SRCR domains are perturbed by SNPs and potential therapeutic reagents. 26146185 6 36 theme binding 725:731 arg1 sites 733:737 the binding sites 721:737 the binding sites on CD6 and CD166 721:754 We characterized the binding sites on CD6 and CD166 and showed that a SNP in CD6 causes glycosylation that hinders the CD6/CD166 interaction. 26146185 2 37 theme cell 311:314 arg1 CD166 334:338 CD166 334:338 CD166 334:338 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 37 theme cell 311:314 arg1 receptor 324:331 another cell surface receptor 303:331 another cell surface receptor 303:331 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 5 38 theme consecutive 645:655 arg1 domains 662:668 consecutive SRCR domains 645:668 consecutive SRCR domains 645:668 This first structure of consecutive SRCR domains reveals a nonlinear organization. 26146185 7 39 theme homophilic 942:951 arg1 interactions 953:964 the heterophilic and homophilic interactions 921:964 interactions 953:964 Native mass spectrometry analysis showed that there is competition between the heterophilic and homophilic interactions. 26146185 5 40 theme SRCR 657:660 arg1 domains 662:668 consecutive SRCR domains 645:668 consecutive SRCR domains 645:668 This first structure of consecutive SRCR domains reveals a nonlinear organization. 26146185 2 41 theme CD6 234:236 arg1 CD6 234:236 CD6 234:236 CD6 234:236 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 2 41 theme CD6 234:236 arg1 domain 224:229 The membrane proximal domain 202:229 The membrane proximal domain of CD6 202:236 The membrane proximal domain of CD6 binds the N-terminal immunoglobulin superfamily (IgSF) domain of another cell surface receptor, CD166, which also engages in homophilic interactions. 26146185 8 42 dep potential 1067:1075 arg1 therapeutic 1077:1087 therapeutic 1077:1087 therapeutic 1077:1087 These data give insight into how interactions of consecutive SRCR domains are perturbed by SNPs and potential therapeutic reagents. 26146185 6 43 from sites 733:737 arg1 CD166 750:754 CD166 750:754 CD166 750:754 We characterized the binding sites on CD6 and CD166 and showed that a SNP in CD6 causes glycosylation that hinders the CD6/CD166 interaction. 26146185 6 43 from sites 733:737 arg1 CD6 742:744 CD6 742:744 CD6 742:744 We characterized the binding sites on CD6 and CD166 and showed that a SNP in CD6 causes glycosylation that hinders the CD6/CD166 interaction. 26146185 7 44 theme spectrometry 858:869 arg1 analysis 871:878 Native mass spectrometry analysis 846:878 Native mass spectrometry analysis 846:878 Native mass spectrometry analysis showed that there is competition between the heterophilic and homophilic interactions. 26146185 4 45 theme domains 569:575 arg1 domains 603:609 two N-terminal domains 588:609 two N-terminal domains of CD166 588:618 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 45 theme domains 569:575 arg1 domains 569:575 the three SRCR domains 554:575 the three SRCR domains of CD6 554:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 45 theme domains 569:575 arg1 CD166 614:618 CD166 614:618 CD166 614:618 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 45 theme domains 569:575 arg1 structures 540:549 the X-ray crystal structures 522:549 the X-ray crystal structures of the three SRCR domains of CD6 522:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 1 46 theme extracellular 115:127 arg1 region 129:134 an extracellular region 112:134 an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains 112:199 CD6 is a transmembrane protein with an extracellular region containing three scavenger receptor cysteine rich (SRCR) domains. 26146185 4 47 theme CD166 614:618 arg1 domains 603:609 two N-terminal domains 588:609 two N-terminal domains of CD166 588:618 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 47 theme CD166 614:618 arg1 domains 569:575 the three SRCR domains 554:575 the three SRCR domains of CD6 554:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 47 theme CD166 614:618 arg1 CD166 614:618 CD166 614:618 CD166 614:618 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 47 theme CD166 614:618 arg1 structures 540:549 the X-ray crystal structures 522:549 the X-ray crystal structures of the three SRCR domains of CD6 522:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 7 48 theme Native 846:851 arg1 analysis 871:878 Native mass spectrometry analysis 846:878 Native mass spectrometry analysis 846:878 Native mass spectrometry analysis showed that there is competition between the heterophilic and homophilic interactions. 26146185 4 49 theme CD6 580:582 arg1 CD6 580:582 CD6 580:582 CD6 580:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 26146185 4 49 theme CD6 580:582 arg1 domains 569:575 the three SRCR domains 554:575 the three SRCR domains of CD6 554:582 We have solved the X-ray crystal structures of the three SRCR domains of CD6 and two N-terminal domains of CD166. 9572875 0 0 theme disulfide 99:107 arg1 linkages 109:116 disulfide linkages 99:116 disulfide linkages 99:116 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 8 1 theme disulfide-linked 1400:1415 arg1 fragments 1425:1433 observed disulfide-linked tryptic fragments 1391:1433 observed disulfide-linked tryptic fragments 1391:1433 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 3 2 theme glycosylated 659:670 arg1 IGFBP-6 672:678 glycosylated IGFBP-6 659:678 glycosylated IGFBP-6 659:678 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 2 3 theme medium-pressure 575:589 arg1 chromatography 591:604 reverse-phase medium-pressure chromatography 561:604 reverse-phase medium-pressure chromatography 561:604 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 4 4 contain contained 762:770 arg2 hexose 824:829 hexose 824:829 hexose 824:829 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 4 4 contain contained 762:770 arg2 N-acetylhexosamine 804:821 N-acetylhexosamine 804:821 N-acetylhexosamine 804:821 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 4 4 contain contained 762:770 arg2 monosaccharides 777:791 8-16 monosaccharides 772:791 8-16 monosaccharides 772:791 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 4 4 contain contained 762:770 arg2 acid 855:858 N-acetylneuraminic acid 836:858 N-acetylneuraminic acid 836:858 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 4 4 contain contained 762:770 arg1 glycoforms 751:760 Major glycoforms 745:760 Major glycoforms 745:760 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 8 5 theme latter 1584:1589 arg1 regions 1591:1597 the latter regions 1580:1597 the latter regions 1580:1597 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 8 5 theme latter 1584:1589 arg1 interconnected 1607:1620 interconnected 1607:1620 interconnected 1607:1620 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 5 6 theme ESMS 980:983 arg1 sequencing 995:1004 Edman sequencing 989:1004 Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography 989:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 6 theme ESMS 980:983 arg1 combination 965:975 a combination 963:975 a combination of ESMS 963:983 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 1 7 theme high-affinity 252:264 arg1 IGFBP-6 303:309 IGFBP-6 303:309 IGFBP-6 303:309 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 7 theme high-affinity 252:264 arg1 IGFBPs 284:289 IGFBPs 284:289 IGFBPs 284:289 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 7 theme high-affinity 252:264 arg1 proteins 274:281 high-affinity binding proteins 252:281 high-affinity binding proteins (IGFBPs) 252:290 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 7 8 theme greater 1229:1235 arg1 resistance 1237:1246 greater resistance 1229:1246 greater resistance to proteolysis 1229:1261 Glycosylated IGFBP-6 exhibited greater resistance to proteolysis by chymotrypsin and trypsin than nonglycosylated IGFBP-6. 9572875 8 9 theme bond 1338:1341 arg1 positions 1343:1351 Native disulfide bond positions 1321:1351 Native disulfide bond positions in IGFBP-6 1321:1362 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 5 10 theme reverse-phase 1040:1052 arg1 chromatography 1075:1088 reverse-phase high-pressure liquid chromatography 1040:1088 reverse-phase high-pressure liquid chromatography 1040:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 4 11 theme N-acetylneuraminic 836:853 arg1 acid 855:858 N-acetylneuraminic acid 836:858 N-acetylneuraminic acid 836:858 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 1 12 theme binding 266:272 arg1 IGFBP-6 303:309 IGFBP-6 303:309 IGFBP-6 303:309 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 12 theme binding 266:272 arg1 IGFBPs 284:289 IGFBPs 284:289 IGFBPs 284:289 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 12 theme binding 266:272 arg1 proteins 274:281 high-affinity binding proteins 252:281 high-affinity binding proteins (IGFBPs) 252:290 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 2 13 theme affinity 533:540 arg1 chromatography 542:555 IGF-II affinity chromatography 526:555 IGF-II affinity chromatography 526:555 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 3 14 gly glycosylated 659:670 arg1 IGFBP-6 672:678 glycosylated IGFBP-6 659:678 glycosylated IGFBP-6 659:678 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 5 15 theme high-pressure 1054:1066 arg1 chromatography 1075:1088 reverse-phase high-pressure liquid chromatography 1040:1088 reverse-phase high-pressure liquid chromatography 1040:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 1 16 theme proteins 274:281 arg1 O-glycosylated 353:366 O-glycosylated 353:366 O-glycosylated 353:366 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 16 theme proteins 274:281 arg1 family 242:247 a family 240:247 a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated 240:366 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 2 17 theme IGF-II 526:531 arg1 chromatography 542:555 IGF-II affinity chromatography 526:555 IGF-II affinity chromatography 526:555 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 5 18 theme fragments 1017:1025 arg1 sequencing 995:1004 Edman sequencing 989:1004 Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography 989:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 18 theme fragments 1017:1025 arg1 combination 965:975 a combination 963:975 a combination of ESMS 963:983 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 19 theme liquid 1068:1073 arg1 chromatography 1075:1088 reverse-phase high-pressure liquid chromatography 1040:1088 reverse-phase high-pressure liquid chromatography 1040:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 2 20 theme nonglycosylated 386:400 arg1 IGFBP-6 420:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 2 21 theme human 414:418 arg1 IGFBP-6 420:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 0 22 theme human 121:125 arg1 factor 147:152 human insulin-like growth factor 121:152 human insulin-like growth factor binding protein 6 121:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 5 23 theme IGFBP-6 884:890 arg1 Thr145 927:932 Thr145 927:932 Thr145 927:932 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 23 theme IGFBP-6 884:890 arg1 Thr146 935:940 Thr146 935:940 Thr146 935:940 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 23 theme IGFBP-6 884:890 arg1 sites 875:879 Glycosylation sites 861:879 Glycosylation sites of IGFBP-6 861:890 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 23 theme IGFBP-6 884:890 arg1 Thr126 911:916 Thr126 911:916 Thr126 911:916 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 23 theme IGFBP-6 884:890 arg1 Ser144 919:924 Ser144 919:924 Ser144 919:924 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 23 theme IGFBP-6 884:890 arg1 Ser152 947:952 Ser152 947:952 Ser152 947:952 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 23 theme IGFBP-6 884:890 arg1 IGFBP-6 884:890 IGFBP-6 884:890 IGFBP-6 884:890 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 7 24 theme Glycosylated 1198:1209 arg1 IGFBP-6 1211:1217 Glycosylated IGFBP-6 1198:1217 Glycosylated IGFBP-6 1198:1217 Glycosylated IGFBP-6 exhibited greater resistance to proteolysis by chymotrypsin and trypsin than nonglycosylated IGFBP-6. 9572875 5 25 theme tryptic 1009:1015 arg1 fragments 1017:1025 tryptic fragments 1009:1025 tryptic fragments separated by reverse-phase high-pressure liquid chromatography 1009:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 1 26 theme insulin-like 188:199 arg1 IGFs 217:220 IGFs 217:220 IGFs 217:220 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 26 theme insulin-like 188:199 arg1 factors 208:214 insulin-like growth factors 188:214 insulin-like growth factors (IGFs) 188:221 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 6 27 contain contained 1167:1175 arg1 others 1160:1165 the others 1156:1165 the others 1156:1165 One oligosaccharide chain contained 5-6 monosaccharides, whereas the others contained 2-4 monosaccharides. 9572875 6 27 contain contained 1167:1175 arg2 monosaccharides 1181:1195 2-4 monosaccharides 1177:1195 2-4 monosaccharides 1177:1195 One oligosaccharide chain contained 5-6 monosaccharides, whereas the others contained 2-4 monosaccharides. 9572875 3 28 gly heterogeneity 702:714 arg1 composition 732:742 carbohydrate composition 719:742 carbohydrate composition 719:742 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 1 29 theme growth 201:206 arg1 IGFs 217:220 IGFs 217:220 IGFs 217:220 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 29 theme growth 201:206 arg1 factors 208:214 insulin-like growth factors 188:214 insulin-like growth factors (IGFs) 188:221 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 2 30 theme ovary 458:462 arg1 cells 464:468 Chinese hamster ovary cells 442:468 Chinese hamster ovary cells 442:468 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 8 31 link disulfide-linked 1400:1415 arg1 fragments 1425:1433 observed disulfide-linked tryptic fragments 1391:1433 observed disulfide-linked tryptic fragments 1391:1433 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 2 32 gly nonglycosylated 386:400 arg1 IGFBP-6 420:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 3 33 theme mass 631:634 arg1 ESMS 650:653 ESMS 650:653 ESMS 650:653 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 3 33 theme mass 631:634 arg1 spectrometry 636:647 Electrospray ionization mass spectrometry 607:647 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 607:678 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 1 34 gly O-glycosylated 353:366 arg1 O-glycosylated 353:366 O-glycosylated 353:366 O-glycosylated 353:366 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 34 gly O-glycosylated 353:366 arg1 family 242:247 a family 240:247 a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated 240:366 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 1 35 theme factors 208:214 arg1 actions 177:183 The actions 173:183 The actions of insulin-like growth factors (IGFs) 173:221 The actions of insulin-like growth factors (IGFs) are modulated by a family of high-affinity binding proteins (IGFBPs), including IGFBP-6, which preferentially binds IGF-II and is O-glycosylated. 9572875 0 36 theme sites 34:38 arg1 Identification 0:13 Identification 0:13 Identification of O-glycosylation sites 0:38 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 0 36 theme sites 34:38 arg1 characterization 52:67 partial characterization 44:67 partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6 44:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 0 37 theme factor 147:152 arg1 structure 85:93 carbohydrate structure 72:93 carbohydrate structure 72:93 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 0 37 theme factor 147:152 arg1 linkages 109:116 disulfide linkages 99:116 disulfide linkages 99:116 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 7 38 gly nonglycosylated 1296:1310 arg1 IGFBP-6 1312:1318 nonglycosylated IGFBP-6 1296:1318 nonglycosylated IGFBP-6 1296:1318 Glycosylated IGFBP-6 exhibited greater resistance to proteolysis by chymotrypsin and trypsin than nonglycosylated IGFBP-6. 9572875 0 39 theme O-glycosylation 18:32 arg1 sites 34:38 O-glycosylation sites 18:38 O-glycosylation sites 18:38 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 0 40 theme growth 140:145 arg1 factor 147:152 human insulin-like growth factor 121:152 human insulin-like growth factor binding protein 6 121:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 8 41 theme previous 1555:1562 arg1 suggestion 1564:1573 a previous suggestion that the latter regions are not interconnected 1553:1620 a previous suggestion that the latter regions are not interconnected 1553:1620 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 8 41 theme previous 1555:1562 arg1 subdomains 1482:1491 two disulfide-linked subdomains 1461:1491 two disulfide-linked subdomains within each of the N- and C-terminal regions 1461:1536 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 3 42 theme Electrospray 607:618 arg1 ESMS 650:653 ESMS 650:653 ESMS 650:653 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 3 42 theme Electrospray 607:618 arg1 spectrometry 636:647 Electrospray ionization mass spectrometry 607:647 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 607:678 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 9 43 theme IGFBP-6 1634:1640 arg1 model 1625:1629 A model 1623:1629 A model of IGFBP-6 1623:1640 A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated. 9572875 0 44 theme partial 44:50 arg1 characterization 52:67 partial characterization 44:67 partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6 44:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 2 45 theme reverse-phase 561:573 arg1 chromatography 591:604 reverse-phase medium-pressure chromatography 561:604 reverse-phase medium-pressure chromatography 561:604 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 3 46 theme carbohydrate 719:730 arg1 composition 732:742 carbohydrate composition 719:742 carbohydrate composition 719:742 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 0 47 theme insulin-like 127:138 arg1 factor 147:152 human insulin-like growth factor 121:152 human insulin-like growth factor binding protein 6 121:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 4 48 theme Major 745:749 arg1 glycoforms 751:760 Major glycoforms 745:760 Major glycoforms 745:760 Major glycoforms contained 8-16 monosaccharides, including N-acetylhexosamine, hexose, and N-acetylneuraminic acid. 9572875 8 49 dep N- 1512:1513 arg1 the 1508:1510 the 1508:1510 the 1508:1510 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 7 50 theme nonglycosylated 1296:1310 arg1 IGFBP-6 1312:1318 nonglycosylated IGFBP-6 1296:1318 nonglycosylated IGFBP-6 1296:1318 Glycosylated IGFBP-6 exhibited greater resistance to proteolysis by chymotrypsin and trypsin than nonglycosylated IGFBP-6. 9572875 9 51 gly O-glycosylated 1730:1743 arg1 region 1714:1719 a central region 1704:1719 a central region which is O-glycosylated 1704:1743 A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated. 9572875 9 51 gly O-glycosylated 1730:1743 arg1 O-glycosylated 1730:1743 O-glycosylated 1730:1743 O-glycosylated 1730:1743 A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated. 9572875 6 52 theme oligosaccharide 1095:1109 arg1 chain 1111:1115 One oligosaccharide chain 1091:1115 One oligosaccharide chain 1091:1115 One oligosaccharide chain contained 5-6 monosaccharides, whereas the others contained 2-4 monosaccharides. 9572875 3 53 theme considerable 689:700 arg1 heterogeneity 702:714 considerable heterogeneity 689:714 considerable heterogeneity of carbohydrate composition 689:742 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 2 54 theme hamster 450:456 arg1 cells 464:468 Chinese hamster ovary cells 442:468 Chinese hamster ovary cells 442:468 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 8 55 theme Native 1321:1326 arg1 positions 1343:1351 Native disulfide bond positions 1321:1351 Native disulfide bond positions in IGFBP-6 1321:1362 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 5 56 theme Glycosylation 861:873 arg1 Thr145 927:932 Thr145 927:932 Thr145 927:932 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 56 theme Glycosylation 861:873 arg1 Thr146 935:940 Thr146 935:940 Thr146 935:940 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 56 theme Glycosylation 861:873 arg1 sites 875:879 Glycosylation sites 861:879 Glycosylation sites of IGFBP-6 861:890 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 56 theme Glycosylation 861:873 arg1 Thr126 911:916 Thr126 911:916 Thr126 911:916 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 56 theme Glycosylation 861:873 arg1 Ser144 919:924 Ser144 919:924 Ser144 919:924 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 56 theme Glycosylation 861:873 arg1 Ser152 947:952 Ser152 947:952 Ser152 947:952 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 5 56 theme Glycosylation 861:873 arg1 IGFBP-6 884:890 IGFBP-6 884:890 IGFBP-6 884:890 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 0 57 gly O-glycosylation 18:32 arg2 sites 34:38 O-glycosylation sites 18:38 O-glycosylation sites 18:38 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 8 58 from positions 1343:1351 arg1 IGFBP-6 1356:1362 IGFBP-6 1356:1362 IGFBP-6 1356:1362 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 5 59 theme Edman 989:993 arg1 sequencing 995:1004 Edman sequencing 989:1004 Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography 989:1088 Glycosylation sites of IGFBP-6 were identified as Thr126, Ser144, Thr145, Thr146, and Ser152 by using a combination of ESMS and Edman sequencing of tryptic fragments separated by reverse-phase high-pressure liquid chromatography. 9572875 3 60 theme ionization 620:629 arg1 ESMS 650:653 ESMS 650:653 ESMS 650:653 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 3 60 theme ionization 620:629 arg1 spectrometry 636:647 Electrospray ionization mass spectrometry 607:647 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 607:678 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 3 61 theme IGFBP-6 672:678 arg1 ESMS 650:653 ESMS 650:653 ESMS 650:653 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 3 61 theme IGFBP-6 672:678 arg1 spectrometry 636:647 Electrospray ionization mass spectrometry 607:647 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 607:678 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 8 62 theme tryptic 1417:1423 arg1 fragments 1425:1433 observed disulfide-linked tryptic fragments 1391:1433 observed disulfide-linked tryptic fragments 1391:1433 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 8 63 theme disulfide-linked 1465:1480 arg1 suggestion 1564:1573 a previous suggestion that the latter regions are not interconnected 1553:1620 a previous suggestion that the latter regions are not interconnected 1553:1620 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 8 63 theme disulfide-linked 1465:1480 arg1 subdomains 1482:1491 two disulfide-linked subdomains 1461:1491 two disulfide-linked subdomains within each of the N- and C-terminal regions 1461:1536 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 8 64 theme C-terminal 1519:1528 arg1 regions 1530:1536 C-terminal regions 1519:1536 C-terminal regions 1519:1536 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 3 65 theme composition 732:742 arg1 heterogeneity 702:714 considerable heterogeneity 689:714 considerable heterogeneity of carbohydrate composition 689:742 Electrospray ionization mass spectrometry (ESMS) of glycosylated IGFBP-6 revealed considerable heterogeneity of carbohydrate composition. 9572875 6 66 contain contained 1117:1125 arg1 chain 1111:1115 One oligosaccharide chain 1091:1115 One oligosaccharide chain 1091:1115 One oligosaccharide chain contained 5-6 monosaccharides, whereas the others contained 2-4 monosaccharides. 9572875 6 66 contain contained 1117:1125 arg2 monosaccharides 1131:1145 5-6 monosaccharides 1127:1145 5-6 monosaccharides 1127:1145 One oligosaccharide chain contained 5-6 monosaccharides, whereas the others contained 2-4 monosaccharides. 9572875 9 67 theme distinct 1670:1677 arg1 domains 1679:1685 these distinct domains 1664:1685 these distinct domains 1664:1685 A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated. 9572875 0 68 theme structure 85:93 arg1 Identification 0:13 Identification 0:13 Identification of O-glycosylation sites 0:38 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 0 68 theme structure 85:93 arg1 characterization 52:67 partial characterization 44:67 partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6 44:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 8 69 theme disulfide 1328:1336 arg1 positions 1343:1351 Native disulfide bond positions 1321:1351 Native disulfide bond positions in IGFBP-6 1321:1362 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 2 70 theme Glycosylated 369:380 arg1 IGFBP-6 420:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 2 71 theme recombinant 402:412 arg1 IGFBP-6 420:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6 369:426 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 9 72 theme central 1706:1712 arg1 region 1714:1719 a central region 1704:1719 a central region which is O-glycosylated 1704:1743 A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated. 9572875 9 72 theme central 1706:1712 arg1 O-glycosylated 1730:1743 O-glycosylated 1730:1743 O-glycosylated 1730:1743 A model of IGFBP-6 is developed in which these distinct domains are separated by a central region which is O-glycosylated. 9572875 0 73 theme carbohydrate 72:83 arg1 structure 85:93 carbohydrate structure 72:93 carbohydrate structure 72:93 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 8 74 link disulfide-linked 1465:1480 arg1 suggestion 1564:1573 a previous suggestion that the latter regions are not interconnected 1553:1620 a previous suggestion that the latter regions are not interconnected 1553:1620 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 8 74 link disulfide-linked 1465:1480 arg1 subdomains 1482:1491 two disulfide-linked subdomains 1461:1491 two disulfide-linked subdomains within each of the N- and C-terminal regions 1461:1536 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 2 75 theme Chinese 442:448 arg1 cells 464:468 Chinese hamster ovary cells 442:468 Chinese hamster ovary cells 442:468 Glycosylated and nonglycosylated recombinant human IGFBP-6, expressed in Chinese hamster ovary cells and Escherichia coli, respectively, were purified using IGF-II affinity chromatography and reverse-phase medium-pressure chromatography. 9572875 8 76 theme observed 1391:1398 arg1 fragments 1425:1433 observed disulfide-linked tryptic fragments 1391:1433 observed disulfide-linked tryptic fragments 1391:1433 Native disulfide bond positions in IGFBP-6 were localized by means of observed disulfide-linked tryptic fragments, revealing that there are two disulfide-linked subdomains within each of the N- and C-terminal regions and confirming a previous suggestion that the latter regions are not interconnected. 9572875 0 77 theme linkages 109:116 arg1 Identification 0:13 Identification 0:13 Identification of O-glycosylation sites 0:38 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 9572875 0 77 theme linkages 109:116 arg1 characterization 52:67 partial characterization 44:67 partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6 44:170 Identification of O-glycosylation sites and partial characterization of carbohydrate structure and disulfide linkages of human insulin-like growth factor binding protein 6. 18491227 0 0 from Asn-633 19:25 arg1 Unglycosylation 0:14 Unglycosylation 0:14 Unglycosylation at Asn-633 0:25 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 2 1 theme N-glycosylation 336:350 arg1 sites 352:356 four potential N-glycosylation sites 321:356 four potential N-glycosylation sites in E-cadherin 321:370 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 2 2 theme N-glycan 461:468 arg1 elimination 446:456 the elimination 442:456 the elimination of N-glycan at Asn-633 442:479 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 2 3 theme potential 326:334 arg1 sites 352:356 four potential N-glycosylation sites 321:356 four potential N-glycosylation sites in E-cadherin 321:370 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 9 4 from Asn-633 1476:1482 arg1 N-glycosylation 1457:1471 N-glycosylation 1457:1471 N-glycosylation at Asn-633 1457:1482 In conclusion, this study revealed that N-glycosylation at Asn-633 is essential for E-cadherin expression, folding and trafficking. 18491227 1 5 theme -dependent 241:250 arg1 adhesion 262:269 Ca(2+)-dependent cell-cell adhesion 235:269 Ca(2+)-dependent cell-cell adhesion 235:269 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 0 6 theme endoplasmic 102:112 arg1 reticulum 114:122 endoplasmic reticulum 102:122 endoplasmic reticulum 102:122 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 2 7 theme previous 277:284 arg1 study 286:290 a previous study 275:290 a previous study 275:290 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 1 8 theme cell-cell 252:260 arg1 adhesion 262:269 Ca(2+)-dependent cell-cell adhesion 235:269 Ca(2+)-dependent cell-cell adhesion 235:269 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 3 9 theme N-glycan 693:700 arg1 Asn-633 705:711 the N-glycan at Asn-633 689:711 the N-glycan at Asn-633 689:711 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 2 10 theme E-cadherin 503:512 arg1 expression 514:523 E-cadherin expression 503:523 E-cadherin expression 503:523 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 3 11 theme at 702:703 arg1 Asn-633 705:711 the N-glycan at Asn-633 689:711 the N-glycan at Asn-633 689:711 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 12 from Asn-633 705:711 arg1 N-glycan 693:700 the N-glycan at Asn-633 689:711 the N-glycan at Asn-633 689:711 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 12 from Asn-633 705:711 arg1 at 702:703 the N-glycan at Asn-633 689:711 the N-glycan at Asn-633 689:711 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 2 13 theme human 401:405 arg1 cells 424:428 human breast carcinoma cells 401:428 human breast carcinoma cells 401:428 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 8 14 theme M4 1302:1303 arg1 E-cadherin 1305:1314 M4 E-cadherin 1302:1314 M4 E-cadherin 1302:1314 Furthermore, we found that N-glycans of M4 E-cadherin were modified in immature high mannose type, suggesting that it could not depart to Golgi apparatus. 18491227 7 15 theme M4 1040:1041 arg1 E-cadherin 1043:1052 M4 E-cadherin 1040:1052 M4 E-cadherin binding to calnexin 1040:1072 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 3 16 from Asn-633 647:653 arg1 folding 727:733 E-cadherin folding 716:733 E-cadherin folding 716:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 7 17 theme E-cadherin 1043:1052 arg1 ration 1030:1035 the ration 1026:1035 the ration of M4 E-cadherin binding to calnexin 1026:1072 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 8 18 theme high 1342:1345 arg1 type 1355:1358 immature high mannose type 1333:1358 immature high mannose type 1333:1358 Furthermore, we found that N-glycans of M4 E-cadherin were modified in immature high mannose type, suggesting that it could not depart to Golgi apparatus. 18491227 7 19 theme other 1124:1128 arg1 variants 1130:1137 other variants 1124:1137 other variants 1124:1137 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 9 20 gly N-glycosylation 1457:1471 arg2 Asn-633 1476:1482 Asn-633 1476:1482 Asn-633 1476:1482 In conclusion, this study revealed that N-glycosylation at Asn-633 is essential for E-cadherin expression, folding and trafficking. 18491227 9 20 gly N-glycosylation 1457:1471 arg1 Asn-633 1476:1482 Asn-633 1476:1482 Asn-633 1476:1482 In conclusion, this study revealed that N-glycosylation at Asn-633 is essential for E-cadherin expression, folding and trafficking. 18491227 9 20 gly N-glycosylation 1457:1471 arg1 E-cadherin 1501:1510 E-cadherin expression 1501:1521 E-cadherin expression 1501:1521 In conclusion, this study revealed that N-glycosylation at Asn-633 is essential for E-cadherin expression, folding and trafficking. 18491227 6 21 theme ERAD 938:941 arg1 substrate 925:933 the substrate 921:933 the substrate of ERAD 921:941 M4 E-cadherin was recognized as the substrate of ERAD and was retro-translocated from ER lumen to cytoplasm by p97. 18491227 6 21 theme ERAD 938:941 arg1 E-cadherin 892:901 M4 E-cadherin 889:901 M4 E-cadherin 889:901 M4 E-cadherin was recognized as the substrate of ERAD and was retro-translocated from ER lumen to cytoplasm by p97. 18491227 9 22 theme E-cadherin 1501:1510 arg1 expression 1512:1521 E-cadherin expression 1501:1521 E-cadherin expression 1501:1521 In conclusion, this study revealed that N-glycosylation at Asn-633 is essential for E-cadherin expression, folding and trafficking. 18491227 3 23 theme E-cadherin 716:725 arg1 folding 727:733 E-cadherin folding 716:733 E-cadherin folding 716:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 0 24 dep and 86:88 arg1 degraded 143:150 degraded 143:150 sequentially degraded by ERAD 130:158 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 1 25 theme single 187:192 arg1 E-cadherin 171:180 The human E-cadherin 161:180 The human E-cadherin 161:180 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 1 25 theme single 187:192 arg1 protein 215:221 a single transmembrane domain protein 185:221 a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion 185:269 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 2 26 from sites 352:356 arg1 E-cadherin 361:370 E-cadherin 361:370 E-cadherin 361:370 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 3 27 theme molecular 581:589 arg1 mechanism 591:599 the molecular mechanism 577:599 the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding 577:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 2 28 theme breast 407:412 arg1 cells 424:428 human breast carcinoma cells 401:428 human breast carcinoma cells 401:428 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 2 29 theme carcinoma 414:422 arg1 cells 424:428 human breast carcinoma cells 401:428 human breast carcinoma cells 401:428 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 3 30 theme Asn-633 705:711 arg1 M4 656:657 M4 656:657 M4 656:657 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 30 theme Asn-633 705:711 arg1 degradation 661:671 degradation 661:671 degradation 661:671 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 30 theme Asn-633 705:711 arg1 Asn-633 647:653 Asn-633 647:653 Asn-633 (M4) 647:658 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 30 theme Asn-633 705:711 arg1 role 681:684 the role 677:684 the role of the N-glycan at Asn-633 in E-cadherin folding 677:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 1 31 theme human 165:169 arg1 E-cadherin 171:180 The human E-cadherin 161:180 The human E-cadherin 161:180 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 1 31 theme human 165:169 arg1 protein 215:221 a single transmembrane domain protein 185:221 a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion 185:269 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 1 32 theme domain 208:213 arg1 E-cadherin 171:180 The human E-cadherin 161:180 The human E-cadherin 161:180 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 1 32 theme domain 208:213 arg1 protein 215:221 a single transmembrane domain protein 185:221 a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion 185:269 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 3 33 from degradation 661:671 arg1 folding 727:733 E-cadherin folding 716:733 E-cadherin folding 716:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 0 34 gly Unglycosylation 0:14 arg1 Asn-633 19:25 Asn-633 19:25 Asn-633 19:25 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 0 34 gly Unglycosylation 0:14 arg2 Asn-633 19:25 Asn-633 19:25 Asn-633 19:25 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 0 35 theme extracellular 32:44 arg1 domain 46:51 extracellular domain 32:51 extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD 32:158 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 0 35 theme extracellular 32:44 arg1 E-cadherin 56:65 E-cadherin 56:65 E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD 56:158 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 2 36 gly N-glycosylation 336:350 arg2 sites 352:356 four potential N-glycosylation sites 321:356 four potential N-glycosylation sites in E-cadherin 321:370 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 2 36 gly N-glycosylation 336:350 arg2 four 321:324 four 321:324 four 321:324 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 8 37 theme immature 1333:1340 arg1 type 1355:1358 immature high mannose type 1333:1358 immature high mannose type 1333:1358 Furthermore, we found that N-glycans of M4 E-cadherin were modified in immature high mannose type, suggesting that it could not depart to Golgi apparatus. 18491227 0 38 theme E-cadherin 56:65 arg1 domain 46:51 extracellular domain 32:51 extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD 32:158 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 0 38 theme E-cadherin 56:65 arg1 E-cadherin 56:65 E-cadherin 56:65 E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD 56:158 Unglycosylation at Asn-633 made extracellular domain of E-cadherin folded incorrectly and arrested in endoplasmic reticulum, then sequentially degraded by ERAD. 18491227 2 39 from Asn-633 473:479 arg1 elimination 446:456 the elimination 442:456 the elimination of N-glycan at Asn-633 442:479 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 8 40 theme E-cadherin 1305:1314 arg1 N-glycans 1289:1297 N-glycans 1289:1297 N-glycans of M4 E-cadherin 1289:1314 Furthermore, we found that N-glycans of M4 E-cadherin were modified in immature high mannose type, suggesting that it could not depart to Golgi apparatus. 18491227 5 41 theme M4 874:875 arg1 E-cadherin 877:886 M4 E-cadherin 874:886 M4 E-cadherin 874:886 Either MG132 or DMM could efficiently block degradation of M4 E-cadherin. 18491227 3 42 from role 681:684 arg1 folding 727:733 E-cadherin folding 716:733 E-cadherin folding 716:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 8 43 theme mannose 1347:1353 arg1 type 1355:1358 immature high mannose type 1333:1358 immature high mannose type 1333:1358 Furthermore, we found that N-glycans of M4 E-cadherin were modified in immature high mannose type, suggesting that it could not depart to Golgi apparatus. 18491227 7 44 theme E-cadherin 1216:1225 arg1 domain 1203:1208 cytoplasmic domain 1191:1208 cytoplasmic domain of M4 E-cadherin 1191:1225 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 7 44 theme E-cadherin 1216:1225 arg1 E-cadherin 1216:1225 M4 E-cadherin 1213:1225 M4 E-cadherin 1213:1225 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 5 45 theme E-cadherin 877:886 arg1 degradation 859:869 degradation 859:869 degradation of M4 E-cadherin 859:886 Either MG132 or DMM could efficiently block degradation of M4 E-cadherin. 18491227 4 46 theme M4 770:771 arg1 E-cadherin 773:782 M4 E-cadherin 770:782 M4 E-cadherin 770:782 We treated cells stably expressed M4 E-cadherin with MG123, DMM, respectively. 18491227 8 47 theme Golgi 1400:1404 arg1 apparatus 1406:1414 Golgi apparatus 1400:1414 Golgi apparatus 1400:1414 Furthermore, we found that N-glycans of M4 E-cadherin were modified in immature high mannose type, suggesting that it could not depart to Golgi apparatus. 18491227 7 48 theme M4 1213:1214 arg1 E-cadherin 1216:1225 M4 E-cadherin 1213:1225 M4 E-cadherin 1213:1225 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 7 49 theme misfolded 1165:1173 arg1 it 1156:1157 it 1156:1157 it 1156:1157 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 7 49 theme misfolded 1165:1173 arg1 protein 1175:1181 a misfolded protein 1163:1181 a misfolded protein 1163:1181 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 1 50 theme transmembrane 194:206 arg1 E-cadherin 171:180 The human E-cadherin 161:180 The human E-cadherin 161:180 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 1 50 theme transmembrane 194:206 arg1 protein 215:221 a single transmembrane domain protein 185:221 a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion 185:269 The human E-cadherin is a single transmembrane domain protein involved in Ca(2+)-dependent cell-cell adhesion. 18491227 7 51 theme cytoplasmic 1191:1201 arg1 domain 1203:1208 cytoplasmic domain 1191:1208 cytoplasmic domain of M4 E-cadherin 1191:1225 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 7 51 theme cytoplasmic 1191:1201 arg1 E-cadherin 1216:1225 M4 E-cadherin 1213:1225 M4 E-cadherin 1213:1225 It was observed that the ration of M4 E-cadherin binding to calnexin was significantly increased compared with that of other variants, suggesting that it was a misfolded protein, though cytoplasmic domain of M4 E-cadherin could associate with beta-catenin. 18491227 3 52 gly N-glycosylation 628:642 arg1 M4 656:657 M4 656:657 M4 656:657 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 52 gly N-glycosylation 628:642 arg1 degradation 661:671 degradation 661:671 degradation 661:671 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 52 gly N-glycosylation 628:642 arg1 Asn-633 647:653 Asn-633 647:653 Asn-633 (M4) 647:658 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 3 52 gly N-glycosylation 628:642 arg1 role 681:684 the role 677:684 the role of the N-glycan at Asn-633 in E-cadherin folding 677:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 6 53 theme M4 889:890 arg1 substrate 925:933 the substrate 921:933 the substrate of ERAD 921:941 M4 E-cadherin was recognized as the substrate of ERAD and was retro-translocated from ER lumen to cytoplasm by p97. 18491227 6 53 theme M4 889:890 arg1 E-cadherin 892:901 M4 E-cadherin 889:901 M4 E-cadherin 889:901 M4 E-cadherin was recognized as the substrate of ERAD and was retro-translocated from ER lumen to cytoplasm by p97. 18491227 3 54 theme E-cadherin 604:613 arg1 mechanism 591:599 the molecular mechanism 577:599 the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding 577:733 In this study we investigated the molecular mechanism of E-cadherin, which lacks N-glycosylation at Asn-633 (M4), degradation and the role of the N-glycan at Asn-633 in E-cadherin folding. 18491227 2 55 used occupied 376:383 arg2 sites 352:356 four potential N-glycosylation sites 321:356 four potential N-glycosylation sites in E-cadherin 321:370 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 2 55 used occupied 376:383 arg2 all 314:316 all 314:316 all 314:316 In a previous study, we demonstrated that all of four potential N-glycosylation sites in E-cadherin are occupied by N-glycans in human breast carcinoma cells in vivo and the elimination of N-glycan at Asn-633 dramatically affected E-cadherin expression and made it degraded. 18491227 6 56 theme ER 975:976 arg1 lumen 978:982 ER lumen 975:982 ER lumen to cytoplasm 975:995 M4 E-cadherin was recognized as the substrate of ERAD and was retro-translocated from ER lumen to cytoplasm by p97. 19141282 6 0 theme IL-7/IL-7Ralpha 880:894 arg1 structures 896:905 The IL-7/IL-7Ralpha structures 876:905 The IL-7/IL-7Ralpha structures 876:905 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 1 1 theme T 223:223 arg1 cells 231:235 T and B cells 223:235 T and B cells 223:235 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 2 2 theme binding 314:320 arg1 constants 322:330 its binding constants 310:330 its binding constants 310:330 We report that the IL-7Ralpha ectodomain uses glycosylation to modulate its binding constants to IL-7, unlike the other receptors in the gamma(c) family. 19141282 6 3 theme IL-7 969:972 arg1 cascade 984:990 the IL-7 signaling cascade 965:990 the IL-7 signaling cascade 965:990 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 3 4 theme enhanced 489:496 arg1 affinity 498:505 the enhanced affinity 485:505 the enhanced affinity 485:505 IL-7 binds glycosylated IL-7Ralpha 300-fold more tightly than unglycosylated IL-7Ralpha, and the enhanced affinity is attributed primarily to an accelerated on rate. 19141282 5 5 with interface 770:778 arg1 IL-7 785:788 IL-7 785:788 IL-7 785:788 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 1 6 theme B 229:229 arg1 cells 231:235 T and B cells 223:235 T and B cells 223:235 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 1 7 theme cells 231:235 arg1 homeostasis 208:218 homeostasis 208:218 homeostasis 208:218 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 1 7 theme cells 231:235 arg1 development 192:202 development 192:202 development 192:202 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 4 8 theme IL-7 583:586 arg1 comparison 569:578 Structural comparison 558:578 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha 558:625 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 3 9 gly unglycosylated 454:467 arg1 IL-7Ralpha 469:478 unglycosylated IL-7Ralpha 454:478 unglycosylated IL-7Ralpha 454:478 IL-7 binds glycosylated IL-7Ralpha 300-fold more tightly than unglycosylated IL-7Ralpha, and the enhanced affinity is attributed primarily to an accelerated on rate. 19141282 4 10 theme IL-7Ralpha 616:625 arg1 forms 607:611 both forms 602:611 both forms of IL-7Ralpha 602:625 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 1 11 theme crucial 139:145 arg1 complex 131:137 a complex 129:137 a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells 129:235 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 6 12 theme signaling 974:982 arg1 cascade 984:990 the IL-7 signaling cascade 965:990 the IL-7 signaling cascade 965:990 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 6 13 theme IL-7-related 1060:1071 arg1 diseases 1073:1080 IL-7-related diseases 1060:1080 IL-7-related diseases 1060:1080 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 5 14 theme binding 762:768 arg1 interface 770:778 the binding interface 758:778 the binding interface with IL-7 758:788 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 4 15 theme binding 691:697 arg1 interface 699:707 the binding interface 687:707 the binding interface 687:707 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 1 16 theme several 150:156 arg1 cascades 168:175 several signaling cascades 150:175 several signaling cascades leading to the development and homeostasis of T and B cells 150:235 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 0 17 theme Structural 0:9 arg1 studies 27:33 Structural and biophysical studies 0:33 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex 0:70 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex. 19141282 5 18 theme expressed 811:819 arg1 mutations 821:829 the expressed mutations 807:829 the expressed mutations 807:829 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 1 19 theme signaling 158:166 arg1 cascades 168:175 several signaling cascades 150:175 several signaling cascades leading to the development and homeostasis of T and B cells 150:235 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 2 20 theme c 381:381 arg1 family 384:389 the gamma(c) family 371:389 the gamma(c) family 371:389 We report that the IL-7Ralpha ectodomain uses glycosylation to modulate its binding constants to IL-7, unlike the other receptors in the gamma(c) family. 19141282 6 21 theme recognition 943:953 arg1 events 955:960 the molecular recognition events 929:960 the molecular recognition events of the IL-7 signaling cascade 929:990 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 5 22 from defects 853:859 arg1 IL-7Ralpha 864:873 IL-7Ralpha 864:873 IL-7Ralpha 864:873 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 2 23 theme IL-7Ralpha 257:266 arg1 ectodomain 268:277 the IL-7Ralpha ectodomain 253:277 the IL-7Ralpha ectodomain 253:277 We report that the IL-7Ralpha ectodomain uses glycosylation to modulate its binding constants to IL-7, unlike the other receptors in the gamma(c) family. 19141282 4 24 from complex 591:597 arg1 comparison 569:578 Structural comparison 558:578 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha 558:625 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 2 25 theme gamma 375:379 arg1 family 384:389 the gamma(c) family 371:389 the gamma(c) family 371:389 We report that the IL-7Ralpha ectodomain uses glycosylation to modulate its binding constants to IL-7, unlike the other receptors in the gamma(c) family. 19141282 0 26 theme biophysical 15:25 arg1 studies 27:33 Structural and biophysical studies 0:33 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex 0:70 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex. 19141282 6 27 theme molecular 933:941 arg1 events 955:960 the molecular recognition events 929:960 the molecular recognition events of the IL-7 signaling cascade 929:990 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 1 28 theme gamma 102:106 arg1 receptor 111:118 the gamma(c) receptor 98:118 the gamma(c) receptor 98:118 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 3 29 theme glycosylated 403:414 arg1 IL-7Ralpha 416:425 glycosylated IL-7Ralpha 403:425 glycosylated IL-7Ralpha 403:425 IL-7 binds glycosylated IL-7Ralpha 300-fold more tightly than unglycosylated IL-7Ralpha, and the enhanced affinity is attributed primarily to an accelerated on rate. 19141282 5 30 theme IL-7Ralpha 732:741 arg1 mutations 719:727 The SCID mutations 710:727 The SCID mutations of IL-7Ralpha 710:741 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 3 31 gly glycosylated 403:414 arg1 IL-7Ralpha 416:425 glycosylated IL-7Ralpha 403:425 glycosylated IL-7Ralpha 403:425 IL-7 binds glycosylated IL-7Ralpha 300-fold more tightly than unglycosylated IL-7Ralpha, and the enhanced affinity is attributed primarily to an accelerated on rate. 19141282 4 32 from IL-7 583:586 arg1 complex 591:597 complex 591:597 complex 591:597 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 2 33 theme other 352:356 arg1 receptors 358:366 the other receptors 348:366 the other receptors in the gamma(c) family 348:389 We report that the IL-7Ralpha ectodomain uses glycosylation to modulate its binding constants to IL-7, unlike the other receptors in the gamma(c) family. 19141282 1 34 dep development 192:202 arg1 the 188:190 the 188:190 the 188:190 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 0 35 theme IL-7/IL-7Ralpha 48:62 arg1 complex 64:70 the human IL-7/IL-7Ralpha complex 38:70 the human IL-7/IL-7Ralpha complex 38:70 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex. 19141282 4 36 from comparison 569:578 arg1 complex 591:597 complex 591:597 complex 591:597 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 0 37 theme human 42:46 arg1 complex 64:70 the human IL-7/IL-7Ralpha complex 38:70 the human IL-7/IL-7Ralpha complex 38:70 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex. 19141282 3 38 theme unglycosylated 454:467 arg1 IL-7Ralpha 469:478 unglycosylated IL-7Ralpha 454:478 unglycosylated IL-7Ralpha 454:478 IL-7 binds glycosylated IL-7Ralpha 300-fold more tightly than unglycosylated IL-7Ralpha, and the enhanced affinity is attributed primarily to an accelerated on rate. 19141282 1 39 theme c 108:108 arg1 receptor 111:118 the gamma(c) receptor 98:118 the gamma(c) receptor 98:118 IL-7 and IL-7Ralpha bind the gamma(c) receptor, forming a complex crucial to several signaling cascades leading to the development and homeostasis of T and B cells. 19141282 5 40 theme SCID 714:717 arg1 mutations 719:727 The SCID mutations 710:727 The SCID mutations of IL-7Ralpha 710:741 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 5 41 theme protein 837:843 arg1 defects 853:859 protein folding defects 837:859 protein folding defects in IL-7Ralpha 837:873 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 4 42 theme Structural 558:567 arg1 comparison 569:578 Structural comparison 558:578 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha 558:625 Structural comparison of IL-7 in complex to both forms of IL-7Ralpha reveals that glycosylation does not participate directly in the binding interface. 19141282 6 43 theme new 1034:1036 arg1 therapeutics 1038:1049 new therapeutics 1034:1049 new therapeutics 1034:1049 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 19141282 2 44 from receptors 358:366 arg1 family 384:389 the gamma(c) family 371:389 the gamma(c) family 371:389 We report that the IL-7Ralpha ectodomain uses glycosylation to modulate its binding constants to IL-7, unlike the other receptors in the gamma(c) family. 19141282 5 45 theme folding 845:851 arg1 defects 853:859 protein folding defects 837:859 protein folding defects in IL-7Ralpha 837:873 The SCID mutations of IL-7Ralpha locate outside the binding interface with IL-7, suggesting that the expressed mutations cause protein folding defects in IL-7Ralpha. 19141282 0 46 theme complex 64:70 arg1 studies 27:33 Structural and biophysical studies 0:33 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex 0:70 Structural and biophysical studies of the human IL-7/IL-7Ralpha complex. 19141282 6 47 theme cascade 984:990 arg1 events 955:960 the molecular recognition events 929:960 the molecular recognition events of the IL-7 signaling cascade 929:990 The IL-7/IL-7Ralpha structures provide a window into the molecular recognition events of the IL-7 signaling cascade and provide sites to target for designing new therapeutics to treat IL-7-related diseases. 10756055 0 0 theme virus 100:104 arg1 type 106:109 human immunodeficiency virus type 1 77:111 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 5 1 theme contact 1147:1153 arg1 sites 1155:1159 the Env-coreceptor contact sites 1128:1159 the Env-coreceptor contact sites 1128:1159 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 0 2 theme immunodeficiency 83:98 arg1 type 106:109 human immunodeficiency virus type 1 77:111 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 4 3 theme HIV-1 895:899 arg1 isolates 901:908 HIV-1 isolates 895:908 HIV-1 isolates 895:908 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 5 4 theme framework 1091:1099 arg1 explanation 968:978 a possible explanation 957:978 a possible explanation for the various observations of restricted virus entry in some cell types 957:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 5 4 theme framework 1091:1099 arg1 understanding 1070:1082 further our understanding 1058:1082 further our understanding of the framework of elements that represent the Env-coreceptor contact sites 1058:1159 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 2 5 theme HIV-1 510:514 arg1 strains 516:522 both X4 and R5 laboratory-adapted and primary HIV-1 strains 464:522 strains 516:522 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 4 6 theme far-reaching 791:802 arg1 implications 804:815 far-reaching implications 791:815 far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 791:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 3 7 theme existing 569:576 arg1 structures 599:608 existing common extracellular structures 569:608 existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction 569:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 1 8 dep receptors 137:145 arg1 receptors 137:145 The chemokine receptors CXCR4 and CCR5 123:160 The chemokine receptors CXCR4 and CCR5 123:160 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 8 dep receptors 137:145 arg1 CCR5 157:160 CCR5 157:160 CCR5 157:160 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 8 dep receptors 137:145 arg1 CXCR4 147:151 CXCR4 147:151 CXCR4 147:151 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 5 9 theme entry 1029:1033 arg1 observations 996:1007 the various observations 984:1007 the various observations of restricted virus entry in some cell types 984:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 0 10 theme type 106:109 arg1 isolates 113:120 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 3 11 theme glycoprotein 736:747 arg1 interaction 755:765 HIV envelope glycoprotein (Env) interaction 723:765 HIV envelope glycoprotein (Env) interaction 723:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 5 12 theme virus 1023:1027 arg1 entry 1029:1033 restricted virus entry 1012:1033 restricted virus entry in some cell types 1012:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 4 13 theme glycosylated 873:884 arg1 CXCR4 886:890 cell type-dependent glycosylated CXCR4 853:890 cell type-dependent glycosylated CXCR4 853:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 0 14 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of CXCR4 0:30 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 1 15 theme principal 170:178 arg1 receptors 137:145 The chemokine receptors CXCR4 and CCR5 123:160 The chemokine receptors CXCR4 and CCR5 123:160 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 15 theme principal 170:178 arg1 coreceptors 180:190 the principal coreceptors 166:190 the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates 166:270 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 3 16 theme HIV 723:725 arg1 Env 750:752 Env 750:752 Env 750:752 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 3 16 theme HIV 723:725 arg1 glycoprotein 736:747 HIV envelope glycoprotein 723:747 HIV envelope glycoprotein (Env) interaction 723:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 4 17 theme type-dependent 858:871 arg1 CXCR4 886:890 cell type-dependent glycosylated CXCR4 853:890 cell type-dependent glycosylated CXCR4 853:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 2 18 from removal 341:347 arg1 CXCR4 388:392 CXCR4 388:392 CXCR4 388:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 5 19 theme restricted 1012:1021 arg1 entry 1029:1033 restricted virus entry 1012:1033 restricted virus entry in some cell types 1012:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 2 20 theme sites 379:383 arg1 removal 341:347 the removal 337:347 the removal of the N-linked glycosylation sites in CXCR4 337:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 3 21 theme elements 675:682 arg1 similarity 651:660 a conserved three-dimensional similarity 621:660 a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 621:700 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 3 22 theme envelope 727:734 arg1 Env 750:752 Env 750:752 Env 750:752 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 3 22 theme envelope 727:734 arg1 glycoprotein 736:747 HIV envelope glycoprotein 723:747 HIV envelope glycoprotein (Env) interaction 723:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 5 23 theme possible 959:966 arg1 explanation 968:978 a possible explanation 957:978 a possible explanation for the various observations of restricted virus entry in some cell types 957:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 5 24 from entry 1029:1033 arg1 types 1048:1052 some cell types 1038:1052 some cell types 1038:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 2 25 theme glycosylation 365:377 arg1 sites 379:383 the N-linked glycosylation sites 352:383 the N-linked glycosylation sites in CXCR4 352:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 0 26 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of CXCR4 0:30 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 1 27 theme isolates 263:270 arg1 infection 196:204 infection 196:204 infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates 196:270 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 2 28 gly glycosylation 365:377 arg2 sites 379:383 the N-linked glycosylation sites 352:383 the N-linked glycosylation sites in CXCR4 352:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 2 29 theme N-linked 356:363 arg1 sites 379:383 the N-linked glycosylation sites 352:383 the N-linked glycosylation sites in CXCR4 352:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 0 30 theme CXCR4 26:30 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of CXCR4 0:30 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 3 31 gly glycoprotein 736:747 arg1 Env 750:752 Env 750:752 Env 750:752 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 3 31 gly glycoprotein 736:747 arg1 glycoprotein 736:747 HIV envelope glycoprotein 723:747 HIV envelope glycoprotein (Env) interaction 723:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 5 32 theme further 1058:1064 arg1 understanding 1070:1082 further our understanding 1058:1082 further our understanding of the framework of elements that represent the Env-coreceptor contact sites 1058:1159 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 4 33 theme cell 853:856 arg1 CXCR4 886:890 cell type-dependent glycosylated CXCR4 853:890 cell type-dependent glycosylated CXCR4 853:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 5 34 theme cell 1043:1046 arg1 types 1048:1052 some cell types 1038:1052 some cell types 1038:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 5 35 theme various 988:994 arg1 observations 996:1007 the various observations 984:1007 the various observations of restricted virus entry in some cell types 984:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 3 36 theme important 665:673 arg1 elements 675:682 important elements 665:682 important elements of CXCR4 and CCR5 665:700 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 2 37 dep observation 320:330 arg1 allows 406:411 allows 406:411 allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains 406:522 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 1 38 theme X4 209:210 arg1 isolates 263:270 X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates 209:270 isolates 263:270 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 0 39 theme coreceptor 38:47 arg1 function 49:56 coreceptor function 38:56 coreceptor function 38:56 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 5 40 from observations 996:1007 arg1 types 1048:1052 some cell types 1038:1052 some cell types 1038:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 2 41 link N-linked 356:363 arg1 sites 379:383 the N-linked glycosylation sites 352:383 the N-linked glycosylation sites in CXCR4 352:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 3 42 theme extracellular 585:597 arg1 structures 599:608 existing common extracellular structures 569:608 existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction 569:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 5 43 from types 1048:1052 arg1 observations 996:1007 the various observations 984:1007 the various observations of restricted virus entry in some cell types 984:1052 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 5 44 theme elements 1104:1111 arg1 framework 1091:1099 the framework 1087:1099 the framework of elements that represent the Env-coreceptor contact sites 1087:1159 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 0 45 gly glycosylation 9:21 arg1 CXCR4 26:30 CXCR4 26:30 CXCR4 26:30 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 1 46 theme R5 216:217 arg1 infection 196:204 infection 196:204 infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates 196:270 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 2 47 theme universal 439:447 arg1 protein 417:423 the protein 413:423 the protein 413:423 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 2 47 theme universal 439:447 arg1 coreceptor 449:458 a universal coreceptor 437:458 a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains 437:522 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 4 48 contain have 786:789 arg1 results 774:780 These results 768:780 These results 768:780 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 4 48 contain have 786:789 arg2 implications 804:815 far-reaching implications 791:815 far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 791:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 4 49 gly glycosylated 873:884 arg1 CXCR4 886:890 cell type-dependent glycosylated CXCR4 853:890 cell type-dependent glycosylated CXCR4 853:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 2 50 theme primary 502:508 arg1 strains 516:522 both X4 and R5 laboratory-adapted and primary HIV-1 strains 464:522 strains 516:522 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 4 51 theme CXCR4 886:890 arg1 recognition 838:848 the differential recognition 821:848 the differential recognition of cell type-dependent glycosylated CXCR4 821:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 3 52 theme common 578:583 arg1 structures 599:608 existing common extracellular structures 569:608 existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction 569:765 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 1 53 theme chemokine 127:135 arg1 receptors 137:145 The chemokine receptors CXCR4 and CCR5 123:160 The chemokine receptors CXCR4 and CCR5 123:160 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 53 theme chemokine 127:135 arg1 CCR5 157:160 CCR5 157:160 CCR5 157:160 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 53 theme chemokine 127:135 arg1 CXCR4 147:151 CXCR4 147:151 CXCR4 147:151 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 53 theme chemokine 127:135 arg1 coreceptors 180:190 the principal coreceptors 166:190 the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates 166:270 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 4 54 theme differential 825:836 arg1 recognition 838:848 the differential recognition 821:848 the differential recognition of cell type-dependent glycosylated CXCR4 821:890 These results may have far-reaching implications for the differential recognition of cell type-dependent glycosylated CXCR4 by HIV-1 isolates and their evolution in vivo. 10756055 3 55 theme CCR5 697:700 arg1 elements 675:682 important elements 665:682 important elements of CXCR4 and CCR5 665:700 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 3 56 theme conserved 623:631 arg1 similarity 651:660 a conserved three-dimensional similarity 621:660 a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 621:700 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 2 57 from sites 379:383 arg1 CXCR4 388:392 CXCR4 388:392 CXCR4 388:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 2 58 theme unexpected 309:318 arg1 observation 320:330 the unexpected observation 305:330 the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains 305:522 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 1 59 theme human 219:223 arg1 type 248:251 human immunodeficiency virus type 1 219:253 human immunodeficiency virus type 1 (HIV-1) 219:261 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 59 theme human 219:223 arg1 HIV-1 256:260 HIV-1 256:260 HIV-1 256:260 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 3 60 theme three-dimensional 633:649 arg1 similarity 651:660 a conserved three-dimensional similarity 621:660 a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 621:700 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 1 61 theme immunodeficiency 225:240 arg1 type 248:251 human immunodeficiency virus type 1 219:253 human immunodeficiency virus type 1 (HIV-1) 219:261 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 61 theme immunodeficiency 225:240 arg1 HIV-1 256:260 HIV-1 256:260 HIV-1 256:260 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 0 62 theme human 77:81 arg1 type 106:109 human immunodeficiency virus type 1 77:111 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 3 63 theme CXCR4 687:691 arg1 elements 675:682 important elements 665:682 important elements of CXCR4 and CCR5 665:700 We hypothesize that this alteration unmasks existing common extracellular structures reflecting a conserved three-dimensional similarity of important elements of CXCR4 and CCR5 that are involved in HIV envelope glycoprotein (Env) interaction. 10756055 2 64 from CXCR4 388:392 arg1 removal 341:347 the removal 337:347 the removal of the N-linked glycosylation sites in CXCR4 337:392 Here we report on the unexpected observation that the removal of the N-linked glycosylation sites in CXCR4 potentially allows the protein to serve as a universal coreceptor for both X4 and R5 laboratory-adapted and primary HIV-1 strains. 10756055 1 65 theme virus 242:246 arg1 type 248:251 human immunodeficiency virus type 1 219:253 human immunodeficiency virus type 1 (HIV-1) 219:261 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 1 65 theme virus 242:246 arg1 HIV-1 256:260 HIV-1 256:260 HIV-1 256:260 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10756055 0 66 theme CCR5-dependent 62:75 arg1 isolates 113:120 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 CCR5-dependent human immunodeficiency virus type 1 isolates 62:120 N-linked glycosylation of CXCR4 masks coreceptor function for CCR5-dependent human immunodeficiency virus type 1 isolates. 10756055 5 67 theme Env-coreceptor 1132:1145 arg1 sites 1155:1159 the Env-coreceptor contact sites 1128:1159 the Env-coreceptor contact sites 1128:1159 They also suggest a possible explanation for the various observations of restricted virus entry in some cell types and further our understanding of the framework of elements that represent the Env-coreceptor contact sites. 10756055 1 68 theme type 248:251 arg1 isolates 263:270 X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates 209:270 isolates 263:270 The chemokine receptors CXCR4 and CCR5 are the principal coreceptors for infection of X4 and R5 human immunodeficiency virus type 1 (HIV-1) isolates, respectively. 10531415 4 0 contain containing 643:652 arg2 mutations 654:662 mutations 654:662 mutations 654:662 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 4 0 contain containing 643:652 arg1 receptors 633:641 D5 receptors 630:641 D5 receptors containing mutations in the three predicted sites of N-linked glycosylation 630:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 7 1 theme mature 1088:1093 arg1 receptors 1095:1103 mature receptors 1088:1103 mature receptors 1088:1103 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 1 2 theme D1 235:236 arg1 subtypes 263:270 the D1 and D5 dopamine receptor subtypes 231:270 the D1 and D5 dopamine receptor subtypes 231:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 6 3 theme site 890:893 arg1 Mutation 865:872 Mutation 865:872 Mutation of a comparable site in the D1 receptor at position N5 865:927 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 7 4 theme binding 1339:1345 arg1 properties 1347:1356 the receptor's ligand binding properties 1317:1356 the receptor's ligand binding properties 1317:1356 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 4 5 theme Polymerase 560:569 arg1 reaction 577:584 Polymerase chain reaction 560:584 Polymerase chain reaction mutagenesis 560:596 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 6 6 from Mutation 865:872 arg1 receptor 905:912 the D1 receptor 898:912 the D1 receptor 898:912 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 3 7 theme plasma 511:516 arg1 localization 527:538 the plasma membrane localization 507:538 the plasma membrane localization of the D1 receptor 507:557 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 1 8 theme D5 242:243 arg1 subtypes 263:270 the D1 and D5 dopamine receptor subtypes 231:270 the D1 and D5 dopamine receptor subtypes 231:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 0 9 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 7 10 theme N-glycosidase 1059:1071 arg1 digestion 1075:1083 N-glycosidase F digestion 1059:1083 N-glycosidase F digestion of mature receptors 1059:1103 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 7 11 theme receptor 1134:1141 arg1 H 1158:1158 (3)H 1155:1158 (3)H 1155:1158 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 7 11 theme receptor 1134:1141 arg1 antagonist 1143:1152 the D5 receptor antagonist [(3)H]SCH23390 1127:1167 the D5 receptor antagonist [(3)H]SCH23390 1127:1167 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 5 12 theme membrane 842:849 arg1 localization 851:862 D5 receptor plasma membrane localization 823:862 D5 receptor plasma membrane localization 823:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 4 13 theme receptors 633:641 arg1 panel 621:625 a panel 619:625 a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation 619:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 2 14 theme transfected 286:296 arg1 cells 306:310 transfected HEK 293 cells 286:310 transfected HEK 293 cells 286:310 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 0 15 theme receptors 96:104 arg1 localization 55:66 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 1 16 theme glycosylation 176:188 arg1 role 159:162 the role 155:162 the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes 155:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 7 17 theme antagonist 1143:1152 arg1 binding 1116:1122 binding 1116:1122 binding of the D5 receptor antagonist [(3)H]SCH23390 1116:1167 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 0 18 theme transfected 109:119 arg1 cells 131:135 transfected mammalian cells 109:135 transfected mammalian cells 109:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 1 19 theme functional 193:202 arg1 expression 217:226 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 6 20 theme position 917:924 arg1 N5 926:927 position N5 917:927 position N5 917:927 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 4 21 theme predicted 677:685 arg1 sites 687:691 the three predicted sites 667:691 the three predicted sites of N-linked glycosylation 667:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 4 21 theme predicted 677:685 arg1 glycosylation 705:717 N-linked glycosylation 696:717 N-linked glycosylation 696:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 7 22 theme Tunicamycin 999:1009 arg1 treatment 1011:1019 Tunicamycin treatment 999:1019 Tunicamycin treatment during receptor biosynthesis 999:1048 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 3 23 theme receptor 550:557 arg1 localization 527:538 the plasma membrane localization 507:538 the plasma membrane localization of the D1 receptor 507:557 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 1 24 from role 159:162 arg1 expression 217:226 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 7 25 theme D5 1267:1268 arg1 receptors 1270:1278 D5 receptors 1267:1278 D5 receptors 1267:1278 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 5 26 theme mutant 734:739 arg1 receptors 741:749 mutant receptors 734:749 mutant receptors 734:749 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 8 27 from requirement 1402:1412 arg1 expression 1468:1477 functional cell surface expression 1444:1477 functional cell surface expression of D1 and D5 dopamine receptors 1444:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 4 28 used used 602:605 arg2 mutagenesis 586:596 Polymerase chain reaction mutagenesis 560:596 Polymerase chain reaction mutagenesis 560:596 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 6 29 theme D1 966:967 arg1 receptor 969:976 the D1 receptor 962:976 the D1 receptor 962:976 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 2 30 with Treatment 273:281 arg1 tunicamycin 317:327 tunicamycin 317:327 tunicamycin 317:327 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 2 30 with Treatment 273:281 arg1 inhibitor 333:341 an inhibitor 330:341 an inhibitor of N-linked oligosaccharide addition 330:378 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 3 31 contain had 490:492 arg1 treatment 480:488 tunicamycin treatment 468:488 tunicamycin treatment 468:488 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 3 31 contain had 490:492 arg2 effect 497:502 no effect 494:502 no effect 494:502 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 1 32 link N-linked 167:174 arg1 glycosylation 176:188 N-linked glycosylation 167:188 N-linked glycosylation 167:188 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 4 33 gly glycosylation 705:717 arg2 sites 687:691 the three predicted sites 667:691 the three predicted sites of N-linked glycosylation 667:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 4 33 gly glycosylation 705:717 arg2 glycosylation 705:717 N-linked glycosylation 696:717 N-linked glycosylation 696:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 2 34 theme receptors 421:429 arg1 localization 402:413 localization 402:413 localization of D5 receptors 402:429 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 8 35 theme cell 1455:1458 arg1 expression 1468:1477 functional cell surface expression 1444:1477 functional cell surface expression of D1 and D5 dopamine receptors 1444:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 4 36 theme glycosylation 705:717 arg1 sites 687:691 the three predicted sites 667:691 the three predicted sites of N-linked glycosylation 667:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 4 36 theme glycosylation 705:717 arg1 glycosylation 705:717 N-linked glycosylation 696:717 N-linked glycosylation 696:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 2 37 theme oligosaccharide 355:369 arg1 addition 371:378 N-linked oligosaccharide addition 346:378 N-linked oligosaccharide addition 346:378 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 0 38 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 1 39 theme dopamine 245:252 arg1 subtypes 263:270 the D1 and D5 dopamine receptor subtypes 231:270 the D1 and D5 dopamine receptor subtypes 231:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 7 40 theme surface 1245:1251 arg1 expression 1253:1262 the cell surface expression 1236:1262 the cell surface expression of D5 receptors 1236:1278 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 5 41 theme major 802:806 arg1 determinant 808:818 the major determinant 798:818 the major determinant of D5 receptor plasma membrane localization 798:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 5 41 theme major 802:806 arg1 glycosylation 766:778 glycosylation 766:778 glycosylation of residue N7 766:792 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 1 42 theme subtypes 263:270 arg1 expression 217:226 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 8 43 theme D1 1482:1483 arg1 expression 1468:1477 functional cell surface expression 1444:1477 functional cell surface expression of D1 and D5 dopamine receptors 1444:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 2 44 link N-linked 346:353 arg1 addition 371:378 N-linked oligosaccharide addition 346:378 N-linked oligosaccharide addition 346:378 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 6 45 contain had 929:931 arg1 Mutation 865:872 Mutation 865:872 Mutation of a comparable site in the D1 receptor at position N5 865:927 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 6 45 contain had 929:931 arg2 effect 936:941 no effect 933:941 no effect 933:941 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 0 46 gly glycosylation 9:21 arg1 D1 83:84 D1 83:84 D1 83:84 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 0 46 gly glycosylation 9:21 arg1 receptors 96:104 dopamine receptors 87:104 dopamine receptors 87:104 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 0 46 gly glycosylation 9:21 arg1 membrane 46:53 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 0 46 gly glycosylation 9:21 arg1 D5 71:72 D5 71:72 D5 71:72 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 0 46 gly glycosylation 9:21 arg1 plasma 39:44 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 6 47 theme cell 985:988 arg1 surface 990:996 the cell surface 981:996 the cell surface 981:996 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 8 48 theme D5 1489:1490 arg1 receptors 1501:1509 D5 dopamine receptors 1489:1509 D5 dopamine receptors 1489:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 5 49 theme residue 783:789 arg1 N7 791:792 residue N7 783:792 residue N7 783:792 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 8 50 link N-linked 1418:1425 arg1 glycosylation 1427:1439 N-linked glycosylation 1418:1439 N-linked glycosylation 1418:1439 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 8 51 theme receptors 1501:1509 arg1 expression 1468:1477 functional cell surface expression 1444:1477 functional cell surface expression of D1 and D5 dopamine receptors 1444:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 6 52 from N5 926:927 arg1 Mutation 865:872 Mutation 865:872 Mutation of a comparable site in the D1 receptor at position N5 865:927 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 5 53 gly glycosylation 766:778 arg1 N7 791:792 residue N7 783:792 residue N7 783:792 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 2 54 theme cells 306:310 arg1 Treatment 273:281 Treatment 273:281 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, 273:379 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 1 55 theme cell 204:207 arg1 expression 217:226 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 4 56 theme chain 571:575 arg1 reaction 577:584 Polymerase chain reaction 560:584 Polymerase chain reaction mutagenesis 560:596 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 6 57 theme D1 902:903 arg1 receptor 905:912 the D1 receptor 898:912 the D1 receptor 898:912 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 3 58 theme tunicamycin 468:478 arg1 treatment 480:488 tunicamycin treatment 468:488 tunicamycin treatment 468:488 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 4 59 link N-linked 696:703 arg1 glycosylation 705:717 N-linked glycosylation 696:717 N-linked glycosylation 696:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 7 60 theme F 1073:1073 arg1 digestion 1075:1083 N-glycosidase F digestion 1059:1083 N-glycosidase F digestion of mature receptors 1059:1103 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 6 61 theme comparable 879:888 arg1 site 890:893 a comparable site 877:893 a comparable site in the D1 receptor 877:912 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 1 62 gly glycosylation 176:188 arg1 subtypes 263:270 the D1 and D5 dopamine receptor subtypes 231:270 the D1 and D5 dopamine receptor subtypes 231:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 1 62 gly glycosylation 176:188 arg1 cell 204:207 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 1 62 gly glycosylation 176:188 arg1 surface 209:215 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 5 63 theme plasma 835:840 arg1 localization 851:862 D5 receptor plasma membrane localization 823:862 D5 receptor plasma membrane localization 823:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 0 64 theme membrane 46:53 arg1 localization 55:66 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 7 65 theme ligand 1332:1337 arg1 properties 1347:1356 the receptor's ligand binding properties 1317:1356 the receptor's ligand binding properties 1317:1356 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 2 66 theme HEK 298:300 arg1 cells 306:310 transfected HEK 293 cells 286:310 transfected HEK 293 cells 286:310 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 5 67 theme localization 851:862 arg1 determinant 808:818 the major determinant 798:818 the major determinant of D5 receptor plasma membrane localization 798:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 5 67 theme localization 851:862 arg1 glycosylation 766:778 glycosylation 766:778 glycosylation of residue N7 766:792 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 4 68 theme D5 630:631 arg1 receptors 633:641 D5 receptors 630:641 D5 receptors containing mutations in the three predicted sites of N-linked glycosylation 630:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 7 69 theme receptor 1028:1035 arg1 biosynthesis 1037:1048 receptor biosynthesis 1028:1048 receptor biosynthesis 1028:1048 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 3 70 theme membrane 518:525 arg1 localization 527:538 the plasma membrane localization 507:538 the plasma membrane localization of the D1 receptor 507:557 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 1 71 theme N-linked 167:174 arg1 glycosylation 176:188 N-linked glycosylation 167:188 N-linked glycosylation 167:188 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 0 72 theme mammalian 121:129 arg1 cells 131:135 transfected mammalian cells 109:135 transfected mammalian cells 109:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 5 73 theme D5 823:824 arg1 localization 851:862 D5 receptor plasma membrane localization 823:862 D5 receptor plasma membrane localization 823:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 3 74 theme D1 547:548 arg1 receptor 550:557 the D1 receptor 543:557 the D1 receptor 543:557 In contrast, tunicamycin treatment had no effect on the plasma membrane localization of the D1 receptor. 10531415 8 75 theme N-linked 1418:1425 arg1 glycosylation 1427:1439 N-linked glycosylation 1418:1439 N-linked glycosylation 1418:1439 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 8 76 theme differential 1389:1400 arg1 requirement 1402:1412 a differential requirement 1387:1412 a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors 1387:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 0 77 theme D5 71:72 arg1 localization 55:66 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 6 78 theme receptor 969:976 arg1 delivery 950:957 the delivery 946:957 the delivery of the D1 receptor to the cell surface 946:996 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 7 79 theme receptors 1270:1278 arg1 expression 1253:1262 the cell surface expression 1236:1262 the cell surface expression of D5 receptors 1236:1278 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 5 80 theme receptors 741:749 arg1 Expression 720:729 Expression 720:729 Expression of mutant receptors 720:749 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 8 81 theme functional 1444:1453 arg1 expression 1468:1477 functional cell surface expression 1444:1477 functional cell surface expression of D1 and D5 dopamine receptors 1444:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 0 82 theme D1 83:84 arg1 localization 55:66 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 0 83 from localization 55:66 arg1 cells 131:135 transfected mammalian cells 109:135 transfected mammalian cells 109:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 0 84 theme dopamine 87:94 arg1 receptors 96:104 dopamine receptors 87:104 dopamine receptors 87:104 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 8 85 theme surface 1460:1466 arg1 expression 1468:1477 functional cell surface expression 1444:1477 functional cell surface expression of D1 and D5 dopamine receptors 1444:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 2 86 theme addition 371:378 arg1 tunicamycin 317:327 tunicamycin 317:327 tunicamycin 317:327 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 2 86 theme addition 371:378 arg1 inhibitor 333:341 an inhibitor 330:341 an inhibitor of N-linked oligosaccharide addition 330:378 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 5 87 theme N7 791:792 arg1 determinant 808:818 the major determinant 798:818 the major determinant of D5 receptor plasma membrane localization 798:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 5 87 theme N7 791:792 arg1 glycosylation 766:778 glycosylation 766:778 glycosylation of residue N7 766:792 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 7 88 theme cell 1240:1243 arg1 expression 1253:1262 the cell surface expression 1236:1262 the cell surface expression of D5 receptors 1236:1278 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 2 89 theme D5 418:419 arg1 receptors 421:429 D5 receptors 418:429 D5 receptors 418:429 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 4 90 theme N-linked 696:703 arg1 glycosylation 705:717 N-linked glycosylation 696:717 N-linked glycosylation 696:717 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 2 91 theme N-linked 346:353 arg1 addition 371:378 N-linked oligosaccharide addition 346:378 N-linked oligosaccharide addition 346:378 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 1 92 theme receptor 254:261 arg1 subtypes 263:270 the D1 and D5 dopamine receptor subtypes 231:270 the D1 and D5 dopamine receptor subtypes 231:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 7 93 dep H 1158:1158 arg1 3 1156:1156 3 1156:1156 3 1156:1156 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 0 94 theme plasma 39:44 arg1 localization 55:66 plasma membrane localization 39:66 plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells 39:135 N-linked glycosylation is required for plasma membrane localization of D5, but not D1, dopamine receptors in transfected mammalian cells. 10531415 7 95 theme key 1224:1226 arg1 role 1228:1231 a key role 1222:1231 a key role 1222:1231 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 2 96 theme plasma 438:443 arg1 membrane 445:452 the plasma membrane 434:452 the plasma membrane 434:452 Treatment of transfected HEK 293 cells with tunicamycin, an inhibitor of N-linked oligosaccharide addition, was found to prevent localization of D5 receptors in the plasma membrane. 10531415 8 97 theme dopamine 1492:1499 arg1 receptors 1501:1509 D5 dopamine receptors 1489:1509 D5 dopamine receptors 1489:1509 Together, our data indicate a differential requirement for N-linked glycosylation in functional cell surface expression of D1 and D5 dopamine receptors. 10531415 7 98 theme receptors 1095:1103 arg1 digestion 1075:1083 N-glycosidase F digestion 1059:1083 N-glycosidase F digestion of mature receptors 1059:1103 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 6 99 from receptor 905:912 arg1 Mutation 865:872 Mutation 865:872 Mutation of a comparable site in the D1 receptor at position N5 865:927 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 7 100 theme oligosaccharide 1192:1206 arg1 moieties 1208:1215 oligosaccharide moieties 1192:1215 oligosaccharide moieties 1192:1215 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 4 101 theme reaction 577:584 arg1 mutagenesis 586:596 Polymerase chain reaction mutagenesis 560:596 Polymerase chain reaction mutagenesis 560:596 Polymerase chain reaction mutagenesis was used to generate a panel of D5 receptors containing mutations in the three predicted sites of N-linked glycosylation. 10531415 1 102 theme surface 209:215 arg1 expression 217:226 functional cell surface expression 193:226 functional cell surface expression of the D1 and D5 dopamine receptor subtypes 193:270 We have analyzed the role of N-linked glycosylation in functional cell surface expression of the D1 and D5 dopamine receptor subtypes. 10531415 5 103 theme receptor 826:833 arg1 localization 851:862 D5 receptor plasma membrane localization 823:862 D5 receptor plasma membrane localization 823:862 Expression of mutant receptors indicated that glycosylation of residue N7 was the major determinant of D5 receptor plasma membrane localization. 10531415 6 104 from site 890:893 arg1 receptor 905:912 the D1 receptor 898:912 the D1 receptor 898:912 Mutation of a comparable site in the D1 receptor at position N5 had no effect on the delivery of the D1 receptor to the cell surface. 10531415 7 105 theme D5 1131:1132 arg1 H 1158:1158 (3)H 1155:1158 (3)H 1155:1158 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 10531415 7 105 theme D5 1131:1132 arg1 antagonist 1143:1152 the D5 receptor antagonist [(3)H]SCH23390 1127:1167 the D5 receptor antagonist [(3)H]SCH23390 1127:1167 Tunicamycin treatment during receptor biosynthesis, but not N-glycosidase F digestion of mature receptors, abrogated binding of the D5 receptor antagonist [(3)H]SCH23390, suggesting that while oligosaccharide moieties play a key role in the cell surface expression of D5 receptors, they do not appear to contribute to the receptor's ligand binding properties. 21569239 10 0 from regulation 1507:1516 arg1 part 1456:1459 part 1456:1459 part of the membrane targeting of F-spondin in its regulation of axon development 1456:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 7 1 dep N- 1020:1021 arg1 the 1016:1018 the 1016:1018 the 1016:1018 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 10 2 theme core 1334:1337 arg1 structure 1339:1347 its core structure 1330:1347 its core structure 1330:1347 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 2 3 theme F-spondin 339:347 arg1 domain 349:354 the F-spondin domain 335:354 the F-spondin domain (FS domain) 335:366 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 2 3 theme F-spondin 339:347 arg1 domain 360:365 FS domain 357:365 FS domain 357:365 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 1 4 theme axon 257:260 arg1 outgrowth 262:270 axon outgrowth 257:270 axon outgrowth 257:270 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 4 5 theme human 623:627 arg1 domain 642:647 human F-spondin FS domain 623:647 human F-spondin FS domain 623:647 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 10 6 theme common 1354:1359 arg1 Ca2+- 1361:1365 a common Ca2+- 1352:1365 a common Ca2+- 1352:1365 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 4 7 theme FS 639:640 arg1 domain 642:647 human F-spondin FS domain 623:647 human F-spondin FS domain 623:647 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 10 8 theme axon 1521:1524 arg1 development 1526:1536 axon development 1521:1536 axon development 1521:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 3 9 located found 518:522 arg1 F-spondins 527:536 F-spondins 527:536 F-spondins 527:536 The FS domain is found in F-spondins, mindins, M-spondin and amphiF-spondin. 21569239 3 9 located found 518:522 arg1 amphiF-spondin 562:575 amphiF-spondin 562:575 amphiF-spondin 562:575 The FS domain is found in F-spondins, mindins, M-spondin and amphiF-spondin. 21569239 3 9 located found 518:522 arg1 M-spondin 548:556 M-spondin 548:556 M-spondin 548:556 The FS domain is found in F-spondins, mindins, M-spondin and amphiF-spondin. 21569239 3 9 located found 518:522 arg2 domain 508:513 The FS domain 501:513 The FS domain 501:513 The FS domain is found in F-spondins, mindins, M-spondin and amphiF-spondin. 21569239 3 9 located found 518:522 arg1 mindins 539:545 mindins 539:545 mindins 539:545 The FS domain is found in F-spondins, mindins, M-spondin and amphiF-spondin. 21569239 1 10 theme extracellular 176:188 arg1 BACKGROUND 136:145 BACKGROUND 136:145 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 136:309 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 10 theme extracellular 176:188 arg1 protein 203:209 a multi-domain extracellular matrix (ECM) protein 161:209 a multi-domain extracellular matrix (ECM) protein 161:209 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 10 theme extracellular 176:188 arg1 F-spondin 148:156 F-spondin 148:156 F-spondin 148:156 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 0 11 theme matrix 120:125 arg1 protein 127:133 an extracellular matrix protein 103:133 an extracellular matrix protein 103:133 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 7 12 link N-linked 1074:1081 arg1 site 1097:1100 a highly conserved N-linked glycosylation site 1055:1100 a highly conserved N-linked glycosylation site 1055:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 13 theme bonds 994:998 arg1 feature 928:934 The unique feature 917:934 The unique feature of F-spondin FS domain 917:957 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 13 theme bonds 994:998 arg1 presence 966:973 the presence 962:973 the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site 962:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 14 theme domain 952:957 arg1 feature 928:934 The unique feature 917:934 The unique feature of F-spondin FS domain 917:957 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 14 theme domain 952:957 arg1 presence 966:973 the presence 962:973 the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site 962:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 6 15 theme overall 880:886 arg1 structures 888:897 their overall structures 874:897 their overall structures 874:897 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 0 16 from variant 92:98 arg1 protein 127:133 an extracellular matrix protein 103:133 an extracellular matrix protein 103:133 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 7 17 theme N-linked 1074:1081 arg1 site 1097:1100 a highly conserved N-linked glycosylation site 1055:1100 a highly conserved N-linked glycosylation site 1055:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 11 18 theme F-spondin 1664:1672 arg1 functions 1651:1659 the functions 1647:1659 the functions of F-spondin 1647:1672 The structural properties of the FS domain revealed in this study pave the way for further exploration into the functions of F-spondin. 21569239 7 19 theme F-spondin 939:947 arg1 domain 952:957 F-spondin FS domain 939:957 F-spondin FS domain 939:957 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 5 20 with variant 717:723 arg1 fold 768:771 an 8-stranded antiparallel β-sandwich fold 730:771 an 8-stranded antiparallel β-sandwich fold 730:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 0 21 theme domain 58:63 arg1 structure 4:12 The structure 0:12 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 0:134 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 5 22 theme β-sandwich 757:766 arg1 fold 768:771 an 8-stranded antiparallel β-sandwich fold 730:771 an 8-stranded antiparallel β-sandwich fold 730:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 2 23 theme commissural 467:477 arg1 axons 479:483 commissural axons 467:483 commissural axons 467:483 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 10 24 theme C2 1385:1386 arg1 domain 1388:1393 lipid-binding C2 domain 1371:1393 lipid-binding C2 domain 1371:1393 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 0 25 theme F-spondin 68:76 arg1 F-spondin 68:76 F-spondin 68:76 F-spondin 68:76 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 0 25 theme F-spondin 68:76 arg1 domain 58:63 the Ca²+-binding, glycosylated F-spondin domain 17:63 domain 58:63 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 9 26 theme F-spondin 1224:1232 arg1 domain 1237:1242 the F-spondin FS domain 1220:1242 the F-spondin FS domain 1220:1242 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 5 27 theme Ca2+-binding 694:705 arg1 variant 717:723 a Ca2+-binding C2 domain variant 692:723 a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold 692:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 9 28 theme domain 1237:1242 arg1 structure 1207:1215 The structure 1203:1215 The structure of the F-spondin FS domain 1203:1242 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 5 29 theme domain 710:715 arg1 variant 717:723 a Ca2+-binding C2 domain variant 692:723 a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold 692:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 5 30 theme antiparallel 744:755 arg1 fold 768:771 an 8-stranded antiparallel β-sandwich fold 730:771 an 8-stranded antiparallel β-sandwich fold 730:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 1 31 theme cell 276:279 arg1 migration 281:289 cell migration 276:289 cell migration 276:289 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 6 32 theme mindin 838:843 arg1 F-spondin 824:832 F-spondin 824:832 F-spondin 824:832 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 32 theme mindin 838:843 arg1 domains 813:819 the FS domains 806:819 the FS domains of F-spondin and mindin 806:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 32 theme mindin 838:843 arg1 mindin 838:843 mindin 838:843 mindin 838:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 10 33 theme FS 1423:1424 arg1 domain 1426:1431 the F-spondin FS domain 1409:1431 the F-spondin FS domain 1409:1431 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 10 33 theme FS 1423:1424 arg1 responsible 1440:1450 responsible 1440:1450 responsible 1440:1450 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 8 34 located found 1130:1134 arg1 mindin 1139:1144 mindin 1139:1144 mindin 1139:1144 The integrin-binding motif found in mindin is not conserved in the F-spondin FS domain. 21569239 8 34 located found 1130:1134 arg2 motif 1124:1128 The integrin-binding motif 1103:1128 The integrin-binding motif found in mindin 1103:1144 The integrin-binding motif found in mindin is not conserved in the F-spondin FS domain. 21569239 7 35 theme site 1097:1100 arg1 feature 928:934 The unique feature 917:934 The unique feature of F-spondin FS domain 917:957 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 35 theme site 1097:1100 arg1 presence 966:973 the presence 962:973 the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site 962:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 9 36 theme ECM 1300:1302 arg1 molecule 1304:1311 the multiple-domain ECM molecule 1280:1311 the multiple-domain ECM molecule 1280:1311 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 0 37 theme Ca²+-binding 21:32 arg1 F-spondin 68:76 F-spondin 68:76 F-spondin 68:76 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 0 37 theme Ca²+-binding 21:32 arg1 domain 58:63 the Ca²+-binding, glycosylated F-spondin domain 17:63 domain 58:63 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 6 38 theme F-spondin 824:832 arg1 F-spondin 824:832 F-spondin 824:832 F-spondin 824:832 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 38 theme F-spondin 824:832 arg1 domains 813:819 the FS domains 806:819 the FS domains of F-spondin and mindin 806:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 38 theme F-spondin 824:832 arg1 mindin 838:843 mindin 838:843 mindin 838:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 0 39 theme glycosylated 35:46 arg1 F-spondin 68:76 F-spondin 68:76 F-spondin 68:76 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 0 39 theme glycosylated 35:46 arg1 domain 58:63 the Ca²+-binding, glycosylated F-spondin domain 17:63 domain 58:63 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 6 40 theme domains 813:819 arg1 identical 863:871 identical 863:871 identical 863:871 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 40 theme domains 813:819 arg1 sequences 793:801 the primary sequences 781:801 the primary sequences of the FS domains of F-spondin and mindin 781:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 10 41 from targeting 1477:1485 arg1 regulation 1507:1516 its regulation 1503:1516 its regulation of axon development 1503:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 10 42 theme targeting 1477:1485 arg1 part 1456:1459 part 1456:1459 part of the membrane targeting of F-spondin in its regulation of axon development 1456:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 2 43 theme FS 357:358 arg1 domain 349:354 the F-spondin domain 335:354 the F-spondin domain (FS domain) 335:366 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 2 43 theme FS 357:358 arg1 domain 360:365 FS domain 357:365 FS domain 357:365 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 0 44 theme F-spondin 48:56 arg1 F-spondin 68:76 F-spondin 68:76 F-spondin 68:76 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 0 44 theme F-spondin 48:56 arg1 domain 58:63 the Ca²+-binding, glycosylated F-spondin domain 17:63 domain 58:63 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 11 45 theme domain 1575:1580 arg1 properties 1554:1563 The structural properties 1539:1563 The structural properties of the FS domain revealed in this study 1539:1603 The structural properties of the FS domain revealed in this study pave the way for further exploration into the functions of F-spondin. 21569239 7 46 theme domain 1044:1049 arg1 N- 1020:1021 N- 1020:1021 N- 1020:1021 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 6 47 theme primary 785:791 arg1 identical 863:871 identical 863:871 identical 863:871 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 47 theme primary 785:791 arg1 sequences 793:801 the primary sequences 781:801 the primary sequences of the FS domains of F-spondin and mindin 781:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 10 48 theme F-spondin 1490:1498 arg1 targeting 1477:1485 the membrane targeting 1464:1485 the membrane targeting of F-spondin in its regulation of axon development 1464:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 3 49 theme FS 505:506 arg1 domain 508:513 The FS domain 501:513 The FS domain 501:513 The FS domain is found in F-spondins, mindins, M-spondin and amphiF-spondin. 21569239 8 50 theme integrin-binding 1107:1122 arg1 motif 1124:1128 The integrin-binding motif 1103:1128 The integrin-binding motif found in mindin 1103:1144 The integrin-binding motif found in mindin is not conserved in the F-spondin FS domain. 21569239 8 51 theme F-spondin 1170:1178 arg1 domain 1183:1188 the F-spondin FS domain 1166:1188 the F-spondin FS domain 1166:1188 The integrin-binding motif found in mindin is not conserved in the F-spondin FS domain. 21569239 10 52 theme development 1526:1536 arg1 regulation 1507:1516 its regulation 1503:1516 its regulation of axon development 1503:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 5 53 theme 8-stranded 733:742 arg1 fold 768:771 an 8-stranded antiparallel β-sandwich fold 730:771 an 8-stranded antiparallel β-sandwich fold 730:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 1 54 theme multi-domain 163:174 arg1 BACKGROUND 136:145 BACKGROUND 136:145 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 136:309 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 54 theme multi-domain 163:174 arg1 protein 203:209 a multi-domain extracellular matrix (ECM) protein 161:209 a multi-domain extracellular matrix (ECM) protein 161:209 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 54 theme multi-domain 163:174 arg1 F-spondin 148:156 F-spondin 148:156 F-spondin 148:156 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 4 55 theme F-spondin 629:637 arg1 domain 642:647 human F-spondin FS domain 623:647 human F-spondin FS domain 623:647 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 0 56 theme extracellular 106:118 arg1 protein 127:133 an extracellular matrix protein 103:133 an extracellular matrix protein 103:133 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 10 57 theme structure 1339:1347 arg1 homology 1318:1325 The homology 1314:1325 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain 1314:1393 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 1 58 theme matrix 190:195 arg1 BACKGROUND 136:145 BACKGROUND 136:145 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 136:309 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 58 theme matrix 190:195 arg1 protein 203:209 a multi-domain extracellular matrix (ECM) protein 161:209 a multi-domain extracellular matrix (ECM) protein 161:209 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 58 theme matrix 190:195 arg1 F-spondin 148:156 F-spondin 148:156 F-spondin 148:156 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 59 theme ECM 198:200 arg1 BACKGROUND 136:145 BACKGROUND 136:145 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 136:309 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 59 theme ECM 198:200 arg1 protein 203:209 a multi-domain extracellular matrix (ECM) protein 161:209 a multi-domain extracellular matrix (ECM) protein 161:209 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 59 theme ECM 198:200 arg1 F-spondin 148:156 F-spondin 148:156 F-spondin 148:156 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 4 60 theme crystal 602:608 arg1 structure 610:618 the crystal structure 598:618 the crystal structure of human F-spondin FS domain at 1.95Å resolution 598:667 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 7 61 theme disulfide 984:992 arg1 bonds 994:998 three disulfide bonds 978:998 three disulfide bonds associated with the N- and C-termini of the domain 978:1049 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 62 theme FS 949:950 arg1 domain 952:957 F-spondin FS domain 939:957 F-spondin FS domain 939:957 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 2 63 theme axons 479:483 arg1 projection 453:462 the projection 449:462 the projection of commissural axons to floor plate 449:498 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 4 64 theme domain 642:647 arg1 structure 610:618 the crystal structure 598:618 the crystal structure of human F-spondin FS domain at 1.95Å resolution 598:667 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 10 65 theme lipid-binding 1371:1383 arg1 domain 1388:1393 lipid-binding C2 domain 1371:1393 lipid-binding C2 domain 1371:1393 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 4 66 theme 1.95Å 652:656 arg1 resolution 658:667 1.95Å resolution 652:667 1.95Å resolution 652:667 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 0 67 theme C2-domain 82:90 arg1 variant 92:98 A C2-domain variant 80:98 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 0:134 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 9 68 theme FS 1234:1235 arg1 domain 1237:1242 the F-spondin FS domain 1220:1242 the F-spondin FS domain 1220:1242 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 7 69 theme unique 921:926 arg1 feature 928:934 The unique feature 917:934 The unique feature of F-spondin FS domain 917:957 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 7 69 theme unique 921:926 arg1 presence 966:973 the presence 962:973 the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site 962:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 10 70 from part 1456:1459 arg1 regulation 1507:1516 its regulation 1503:1516 its regulation of axon development 1503:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 5 71 theme C2 707:708 arg1 variant 717:723 a Ca2+-binding C2 domain variant 692:723 a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold 692:771 The structure reveals a Ca2+-binding C2 domain variant with an 8-stranded antiparallel β-sandwich fold. 21569239 2 72 theme floor 488:492 arg1 plate 494:498 floor plate 488:498 floor plate 488:498 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 9 73 theme structural 1258:1267 arg1 studies 1269:1275 the structural studies 1254:1275 the structural studies of the multiple-domain ECM molecule 1254:1311 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 2 74 theme cell 424:427 arg1 membrane 429:436 the cell membrane 420:436 the cell membrane 420:436 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 8 75 theme FS 1180:1181 arg1 domain 1183:1188 the F-spondin FS domain 1166:1188 the F-spondin FS domain 1166:1188 The integrin-binding motif found in mindin is not conserved in the F-spondin FS domain. 21569239 0 76 gly glycosylated 35:46 arg1 F-spondin 68:76 F-spondin 68:76 F-spondin 68:76 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 0 76 gly glycosylated 35:46 arg1 domain 58:63 the Ca²+-binding, glycosylated F-spondin domain 17:63 domain 58:63 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 7 77 theme glycosylation 1083:1095 arg1 site 1097:1100 a highly conserved N-linked glycosylation site 1055:1100 a highly conserved N-linked glycosylation site 1055:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 9 78 theme multiple-domain 1284:1298 arg1 molecule 1304:1311 the multiple-domain ECM molecule 1280:1311 the multiple-domain ECM molecule 1280:1311 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 10 79 theme F-spondin 1413:1421 arg1 domain 1426:1431 the F-spondin FS domain 1409:1431 the F-spondin FS domain 1409:1431 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 10 79 theme F-spondin 1413:1421 arg1 responsible 1440:1450 responsible 1440:1450 responsible 1440:1450 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 7 80 gly glycosylation 1083:1095 arg2 site 1097:1100 a highly conserved N-linked glycosylation site 1055:1100 a highly conserved N-linked glycosylation site 1055:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 2 81 theme proteolytic 379:389 arg1 fragment 391:398 a proteolytic fragment 377:398 a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate 377:498 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 7 82 theme conserved 1064:1072 arg1 site 1097:1100 a highly conserved N-linked glycosylation site 1055:1100 a highly conserved N-linked glycosylation site 1055:1100 The unique feature of F-spondin FS domain is the presence of three disulfide bonds associated with the N- and C-termini of the domain and a highly conserved N-linked glycosylation site. 21569239 9 83 theme molecule 1304:1311 arg1 studies 1269:1275 the structural studies 1254:1275 the structural studies of the multiple-domain ECM molecule 1254:1311 CONCLUSION: The structure of the F-spondin FS domain completes the structural studies of the multiple-domain ECM molecule. 21569239 6 84 theme FS 810:811 arg1 F-spondin 824:832 F-spondin 824:832 F-spondin 824:832 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 84 theme FS 810:811 arg1 domains 813:819 the FS domains 806:819 the FS domains of F-spondin and mindin 806:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 6 84 theme FS 810:811 arg1 mindin 838:843 mindin 838:843 mindin 838:843 Though the primary sequences of the FS domains of F-spondin and mindin are less than 36% identical, their overall structures are very similar. 21569239 11 85 theme further 1622:1628 arg1 exploration 1630:1640 further exploration 1622:1640 further exploration into the functions of F-spondin 1622:1672 The structural properties of the FS domain revealed in this study pave the way for further exploration into the functions of F-spondin. 21569239 11 86 theme structural 1543:1552 arg1 properties 1554:1563 The structural properties 1539:1563 The structural properties of the FS domain revealed in this study 1539:1603 The structural properties of the FS domain revealed in this study pave the way for further exploration into the functions of F-spondin. 21569239 2 87 theme reelin_N 315:322 arg1 domain 324:329 The reelin_N domain 311:329 The reelin_N domain 311:329 The reelin_N domain and the F-spondin domain (FS domain) comprise a proteolytic fragment that interacts with the cell membrane and guides the projection of commissural axons to floor plate. 21569239 0 88 dep structure 4:12 arg1 variant 92:98 A C2-domain variant 80:98 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 0:134 The structure of the Ca²+-binding, glycosylated F-spondin domain of F-spondin - A C2-domain variant in an extracellular matrix protein. 21569239 10 89 theme membrane 1468:1475 arg1 targeting 1477:1485 the membrane targeting 1464:1485 the membrane targeting of F-spondin in its regulation of axon development 1464:1536 The homology of its core structure to a common Ca2+- and lipid-binding C2 domain suggests that the F-spondin FS domain may be responsible for part of the membrane targeting of F-spondin in its regulation of axon development. 21569239 11 90 theme FS 1572:1573 arg1 domain 1575:1580 the FS domain 1568:1580 the FS domain revealed in this study 1568:1603 The structural properties of the FS domain revealed in this study pave the way for further exploration into the functions of F-spondin. 21569239 4 91 from resolution 658:667 arg1 structure 610:618 the crystal structure 598:618 the crystal structure of human F-spondin FS domain at 1.95Å resolution 598:667 RESULTS: We present the crystal structure of human F-spondin FS domain at 1.95Å resolution. 21569239 1 92 theme contact-repellent 217:233 arg1 BACKGROUND 136:145 BACKGROUND 136:145 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 136:309 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 21569239 1 92 theme contact-repellent 217:233 arg1 molecule 235:242 a contact-repellent molecule 215:242 a contact-repellent molecule 215:242 BACKGROUND: F-spondin is a multi-domain extracellular matrix (ECM) protein and a contact-repellent molecule that directs axon outgrowth and cell migration during development. 9535843 6 0 from site 927:930 arg1 position 935:942 position 418 935:946 position 418 935:946 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 6 1 theme glycosylation 913:925 arg1 site 927:930 a unique glycosylation site 904:930 a unique glycosylation site 904:930 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 7 2 theme additional 1098:1107 arg1 loops 1123:1127 two additional extracellular loops 1094:1127 two additional extracellular loops between H4 and H5, and H6 and H7 1094:1160 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 7 3 theme site 1027:1030 arg1 insertion 1036:1044 insertion 1036:1044 insertion of ten separate glycosylation sites 1036:1080 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 7 3 theme site 1027:1030 arg1 removal 1011:1017 removal 1011:1017 removal of this site 1011:1030 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 6 4 theme unique 906:911 arg1 site 927:930 a unique glycosylation site 904:930 a unique glycosylation site 904:930 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 7 5 theme extracellular 1109:1121 arg1 loops 1123:1127 two additional extracellular loops 1094:1127 two additional extracellular loops between H4 and H5, and H6 and H7 1094:1160 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 4 6 theme experimental 697:708 arg1 evidence 710:717 experimental evidence 697:717 experimental evidence supporting this model 697:739 However, experimental evidence supporting this model is missing. 9535843 6 7 from presence 892:899 arg1 position 935:942 position 418 935:946 position 418 935:946 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 9 8 theme hydrophobic 1258:1268 arg1 intracellular 1329:1341 intracellular 1329:1341 intracellular 1329:1341 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 9 8 theme hydrophobic 1258:1268 arg1 region 1270:1275 the first hydrophobic region 1248:1275 the first hydrophobic region of Trp rather than being a transmembrane segment 1248:1324 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 3 9 theme transmembrane 571:583 arg1 segments 590:597 six transmembrane (TM) segments 567:597 six transmembrane (TM) segments flanked by intracellular N and C termini 567:638 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 1 10 theme phospholipase 275:287 arg1 pathway 291:297 the phospholipase C pathway 271:297 the phospholipase C pathway 271:297 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 0 11 theme glycosylation-scanning 79:100 arg1 mutagenesis 102:112 glycosylation-scanning mutagenesis 79:112 glycosylation-scanning mutagenesis 79:112 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 2 12 theme Kyte-Doolittle 300:313 arg1 analysis 315:322 Kyte-Doolittle analysis 300:322 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins 300:365 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 1 13 theme C 289:289 arg1 pathway 291:297 the phospholipase C pathway 271:297 the phospholipase C pathway 271:297 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 0 14 theme epitope 118:124 arg1 immunocytochemistry 126:144 epitope immunocytochemistry 118:144 epitope immunocytochemistry 118:144 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 3 15 theme alpha1 525:530 arg1 subunits 532:539 voltage-gated calcium channel alpha1 subunits 495:539 voltage-gated calcium channel alpha1 subunits 495:539 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 10 16 theme region 1440:1445 arg1 center 1412:1417 the center 1408:1417 the center of the putative pore region 1408:1445 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 3 17 theme voltage-gated 495:507 arg1 channel 517:523 voltage-gated calcium channel 495:523 voltage-gated calcium channel alpha1 subunits 495:539 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 3 18 dep N 624:624 arg1 termini 632:638 termini 632:638 termini 632:638 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 8 19 theme transmembrane 1202:1214 arg1 segments 1216:1223 six transmembrane segments 1198:1223 six transmembrane segments formed of H2-H7 1198:1239 This demonstrated the existence of six transmembrane segments formed of H2-H7. 9535843 5 20 theme human 759:763 arg1 Trp 765:767 human Trp 3 759:769 human Trp 3 759:769 Using human Trp 3 to test Trp topology, we now confirm the intracellular nature of the termini by immunocytochemistry. 9535843 9 21 theme Trp 1280:1282 arg1 intracellular 1329:1341 intracellular 1329:1341 intracellular 1329:1341 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 9 21 theme Trp 1280:1282 arg1 region 1270:1275 the first hydrophobic region 1248:1275 the first hydrophobic region of Trp rather than being a transmembrane segment 1248:1324 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 5 22 theme Trp 779:781 arg1 topology 783:790 Trp topology 779:790 Trp topology 779:790 Using human Trp 3 to test Trp topology, we now confirm the intracellular nature of the termini by immunocytochemistry. 9535843 3 23 theme limited 464:470 arg1 similarity 481:490 A limited sequence similarity 462:490 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits 462:539 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 0 24 theme membrane 4:11 arg1 topology 13:20 The membrane topology 0:20 The membrane topology of human transient receptor potential 3 as 0:63 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 1 25 theme receptor 157:164 arg1 Trp 177:179 Trp 177:179 Trp 177:179 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 1 25 theme receptor 157:164 arg1 potential 166:174 Transient receptor potential 147:174 Transient receptor potential (Trp) proteins 147:189 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 7 26 theme glycosylation 1062:1074 arg1 sites 1076:1080 ten separate glycosylation sites 1049:1080 ten separate glycosylation sites 1049:1080 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 6 27 theme extracellular 967:979 arg1 loop 981:984 one extracellular loop 963:984 one extracellular loop between H2 and H3 963:1002 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 2 28 theme acid 337:340 arg1 sequence 342:349 the amino acid sequence 327:349 the amino acid sequence of Trp proteins 327:365 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 1 29 theme ion 196:198 arg1 channels 200:207 ion channels 196:207 ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway 196:297 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 9 30 theme first 1252:1256 arg1 intracellular 1329:1341 intracellular 1329:1341 intracellular 1329:1341 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 9 30 theme first 1252:1256 arg1 region 1270:1275 the first hydrophobic region 1248:1275 the first hydrophobic region of Trp rather than being a transmembrane segment 1248:1324 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 1 31 theme pathway 291:297 arg1 stimulation 256:266 stimulation 256:266 stimulation of the phospholipase C pathway 256:297 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 7 32 theme sites 1076:1080 arg1 insertion 1036:1044 insertion 1036:1044 insertion of ten separate glycosylation sites 1036:1080 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 7 32 theme sites 1076:1080 arg1 removal 1011:1017 removal 1011:1017 removal of this site 1011:1030 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 2 33 theme amino 331:335 arg1 sequence 342:349 the amino acid sequence 327:349 the amino acid sequence of Trp proteins 327:365 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 7 34 gly glycosylation 1062:1074 arg2 ten 1049:1051 ten 1049:1051 ten 1049:1051 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 7 34 gly glycosylation 1062:1074 arg2 sites 1076:1080 ten separate glycosylation sites 1049:1080 ten separate glycosylation sites 1049:1080 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 3 35 theme TM 586:587 arg1 segments 590:597 six transmembrane (TM) segments 567:597 six transmembrane (TM) segments flanked by intracellular N and C termini 567:638 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 2 36 theme transmembrane 438:450 arg1 segments 452:459 forming transmembrane segments 430:459 forming transmembrane segments 430:459 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 2 37 theme segments 452:459 arg1 potential 417:425 potential 417:425 potential of forming transmembrane segments 417:459 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 7 38 theme separate 1053:1060 arg1 sites 1076:1080 ten separate glycosylation sites 1049:1080 ten separate glycosylation sites 1049:1080 After removal of this site and insertion of ten separate glycosylation sites, we defined two additional extracellular loops between H4 and H5, and H6 and H7. 9535843 10 39 gly glycosylated 1451:1462 arg1 site 1393:1396 A site 1391:1396 A site placed in the center of the putative pore region 1391:1445 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 2 40 theme forming 430:436 arg1 segments 452:459 forming transmembrane segments 430:459 forming transmembrane segments 430:459 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 0 41 theme transient 31:39 arg1 potential 50:58 human transient receptor potential 3 25:60 human transient receptor potential 3 25:60 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 9 42 theme transmembrane 1304:1316 arg1 segment 1318:1324 a transmembrane segment 1302:1324 a transmembrane segment 1302:1324 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 8 43 theme segments 1216:1223 arg1 existence 1185:1193 the existence 1181:1193 the existence of six transmembrane segments formed of H2-H7 1181:1239 This demonstrated the existence of six transmembrane segments formed of H2-H7. 9535843 3 44 theme sequence 472:479 arg1 similarity 481:490 A limited sequence similarity 462:490 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits 462:539 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 0 45 theme human 25:29 arg1 potential 50:58 human transient receptor potential 3 25:60 human transient receptor potential 3 25:60 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 10 46 theme channel 1571:1577 arg1 assembly 1579:1586 channel assembly 1571:1586 channel assembly 1571:1586 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 3 47 theme putative 646:653 arg1 region 660:665 a putative pore region 644:665 a putative pore region between TM5 and TM6 644:685 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 1 48 theme calcium 227:233 arg1 entry 235:239 the calcium entry 223:239 the calcium entry observed after stimulation of the phospholipase C pathway 223:297 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 0 49 theme potential 50:58 arg1 topology 13:20 The membrane topology 0:20 The membrane topology of human transient receptor potential 3 as 0:63 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 10 50 theme putative 1426:1433 arg1 region 1440:1445 the putative pore region 1422:1445 the putative pore region 1422:1445 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 6 51 from position 935:942 arg1 presence 892:899 presence 892:899 presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3 892:1002 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 3 52 theme pore 655:658 arg1 region 660:665 a putative pore region 644:665 a putative pore region between TM5 and TM6 644:685 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 2 53 theme proteins 358:365 arg1 sequence 342:349 the amino acid sequence 327:349 the amino acid sequence of Trp proteins 327:365 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 0 54 theme receptor 41:48 arg1 potential 50:58 human transient receptor potential 3 25:60 human transient receptor potential 3 25:60 The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. 9535843 3 55 theme intracellular 610:622 arg1 N 624:624 intracellular N 610:624 intracellular N 610:624 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 2 56 theme Trp 354:356 arg1 proteins 358:365 Trp proteins 354:365 Trp proteins 354:365 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 10 57 theme pore 1435:1438 arg1 region 1440:1445 the putative pore region 1422:1445 the putative pore region 1422:1445 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 3 58 theme segments 590:597 arg1 region 660:665 a putative pore region 644:665 a putative pore region between TM5 and TM6 644:685 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 3 58 theme segments 590:597 arg1 prediction 553:562 the prediction 549:562 the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini 549:638 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 10 59 theme assembly 1579:1586 arg1 stage 1562:1566 a late stage 1555:1566 a late stage of channel assembly 1555:1586 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 1 60 theme Transient 147:155 arg1 Trp 177:179 Trp 177:179 Trp 177:179 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 1 60 theme Transient 147:155 arg1 potential 166:174 Transient receptor potential 147:174 Transient receptor potential (Trp) proteins 147:189 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 2 61 theme sequence 342:349 arg1 analysis 315:322 Kyte-Doolittle analysis 300:322 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins 300:365 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 3 62 theme calcium 509:515 arg1 channel 517:523 voltage-gated calcium channel 495:523 voltage-gated calcium channel alpha1 subunits 495:539 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 10 63 theme late 1557:1560 arg1 stage 1562:1566 a late stage 1555:1566 a late stage of channel assembly 1555:1586 A site placed in the center of the putative pore region was glycosylated, suggesting that this region may have been luminal and was reinserted into the membrane at a late stage of channel assembly. 9535843 5 64 theme termini 840:846 arg1 nature 826:831 the intracellular nature 808:831 the intracellular nature of the termini by immunocytochemistry 808:869 Using human Trp 3 to test Trp topology, we now confirm the intracellular nature of the termini by immunocytochemistry. 9535843 1 65 theme potential 166:174 arg1 proteins 182:189 Transient receptor potential (Trp) proteins 147:189 Transient receptor potential (Trp) proteins 147:189 Transient receptor potential (Trp) proteins form ion channels implicated in the calcium entry observed after stimulation of the phospholipase C pathway. 9535843 3 66 theme channel 517:523 arg1 subunits 532:539 voltage-gated calcium channel alpha1 subunits 495:539 voltage-gated calcium channel alpha1 subunits 495:539 A limited sequence similarity to voltage-gated calcium channel alpha1 subunits lead to the prediction of six transmembrane (TM) segments flanked by intracellular N and C termini and a putative pore region between TM5 and TM6. 9535843 9 67 theme protein-protein 1361:1375 arg1 interactions 1377:1388 protein-protein interactions 1361:1388 protein-protein interactions 1361:1388 Thus, the first hydrophobic region of Trp rather than being a transmembrane segment is intracellular and available for protein-protein interactions. 9535843 6 68 theme site 927:930 arg1 presence 892:899 presence 892:899 presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3 892:1002 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 9535843 5 69 theme intracellular 812:824 arg1 nature 826:831 the intracellular nature 808:831 the intracellular nature of the termini by immunocytochemistry 808:869 Using human Trp 3 to test Trp topology, we now confirm the intracellular nature of the termini by immunocytochemistry. 9535843 2 70 theme hydrophobic 384:394 arg1 H1-H7 405:409 H1-H7 405:409 H1-H7 405:409 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 2 70 theme hydrophobic 384:394 arg1 regions 396:402 seven hydrophobic regions 378:402 seven hydrophobic regions (H1-H7) 378:410 Kyte-Doolittle analysis of the amino acid sequence of Trp proteins identifies seven hydrophobic regions (H1-H7) with potential of forming transmembrane segments. 9535843 6 71 gly glycosylation 913:925 arg2 site 927:930 a unique glycosylation site 904:930 a unique glycosylation site 904:930 We also demonstrate presence of a unique glycosylation site in position 418, which defines one extracellular loop between H2 and H3. 12527193 7 0 dep Crumbs 970:975 arg1 contrast 947:954 contrast 947:954 contrast 947:954 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 7 1 theme short 1003:1007 arg1 domain 1023:1028 a very short extracellular domain 996:1028 a very short extracellular domain 996:1028 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 8 2 theme extracellular 1213:1225 arg1 domains 1227:1233 divergent extracellular domains 1203:1233 divergent extracellular domains 1203:1233 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 4 3 theme eye 546:548 arg1 disorders 550:558 the eye disorders 542:558 the eye disorders 542:558 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 4 3 theme eye 546:548 arg1 pigmentosa 571:580 retinitis pigmentosa 561:580 retinitis pigmentosa 561:580 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 4 3 theme eye 546:548 arg1 amaurosis 603:611 Leber congenital amaurosis 586:611 Leber congenital amaurosis 586:611 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 6 4 theme current 842:848 arg1 report 850:855 the current report 838:855 the current report 838:855 In the current report we have cloned a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3). 12527193 7 5 theme extracellular 1009:1021 arg1 domain 1023:1028 a very short extracellular domain 996:1028 a very short extracellular domain 996:1028 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 2 6 theme polarity 341:348 arg1 disruption 322:331 disruption 322:331 disruption of cell polarity 322:348 Overexpression of Crumbs in Drosophila epithelia expands the apical surface and leads to disruption of cell polarity. 12527193 9 7 link linked 1360:1365 arg1 site 1381:1384 the conserved N linked glycosylation site 1344:1384 the conserved N linked glycosylation site 1344:1384 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 9 7 link linked 1360:1365 arg1 necessary 1408:1416 necessary 1408:1416 necessary 1408:1416 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 3 8 theme Drosophila 351:360 arg1 Crumbs 362:367 Drosophila Crumbs 351:367 Drosophila Crumbs 351:367 Drosophila Crumbs also interacts with two other polarity genes, Stardust and Discs Lost. 12527193 4 9 theme Crumbs 500:505 arg1 orthologue 475:484 a human orthologue 467:484 a human orthologue 467:484 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 9 10 theme CRB3 1422:1425 arg1 targeting 1434:1442 CRB3 apical targeting 1422:1442 CRB3 apical targeting 1422:1442 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 9 11 theme apical 1427:1432 arg1 targeting 1434:1442 CRB3 apical targeting 1422:1442 CRB3 apical targeting 1422:1442 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 6 12 theme human 897:901 arg1 paralogue 903:911 a human paralogue 895:911 a human paralogue of CRB1 called Crumbs3 (CRB3) 895:941 In the current report we have cloned a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3). 12527193 5 13 theme Stardust 705:712 arg1 orthologues 690:700 mammalian orthologues 680:700 mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively 680:832 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 10 14 theme tight 1574:1578 arg1 junction 1580:1587 the tight junction 1570:1587 the tight junction 1570:1587 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 5 15 theme Pals1 781:785 arg1 PATJ 814:817 PATJ 814:817 PATJ 814:817 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 5 15 theme Pals1 781:785 arg1 junction 804:811 Pals1 associated tight junction 781:811 Pals1 associated tight junction (PATJ) 781:818 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 4 16 theme congenital 592:601 arg1 disorders 550:558 the eye disorders 542:558 the eye disorders 542:558 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 4 16 theme congenital 592:601 arg1 amaurosis 603:611 Leber congenital amaurosis 586:611 Leber congenital amaurosis 586:611 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 9 17 theme tight 1324:1328 arg1 junctions 1330:1338 tight junctions 1324:1338 tight junctions 1324:1338 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 7 18 theme intracellular 1073:1085 arg1 domain 1087:1092 a conserved intracellular domain 1061:1092 a conserved intracellular domain that allows it to complex with Pals1 and PATJ 1061:1138 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 9 19 theme apical 1305:1310 arg1 surface 1312:1318 the apical surface 1301:1318 the apical surface 1301:1318 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 6 20 theme CRB1 916:919 arg1 paralogue 903:911 a human paralogue 895:911 a human paralogue of CRB1 called Crumbs3 (CRB3) 895:941 In the current report we have cloned a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3). 12527193 2 21 theme Drosophila 261:270 arg1 epithelia 272:280 Drosophila epithelia 261:280 Drosophila epithelia 261:280 Overexpression of Crumbs in Drosophila epithelia expands the apical surface and leads to disruption of cell polarity. 12527193 7 22 theme conserved 1063:1071 arg1 domain 1087:1092 a conserved intracellular domain 1061:1092 a conserved intracellular domain that allows it to complex with Pals1 and PATJ 1061:1138 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 10 23 theme Crumbs 1482:1487 arg1 family 1497:1502 the Crumbs protein family 1478:1502 the Crumbs protein family 1478:1502 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 4 24 theme human 469:473 arg1 orthologue 475:484 a human orthologue 467:484 a human orthologue 467:484 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 8 25 theme N-glycosylation 1258:1272 arg1 site 1274:1277 a conserved N-glycosylation site 1246:1277 a conserved N-glycosylation site 1246:1277 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 1 26 theme important 159:167 arg1 role 169:172 an important role 156:172 an important role 156:172 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 0 27 theme Mammalian 0:8 arg1 protein 43:49 a small transmembrane protein 21:49 a small transmembrane protein linked to protein associated with Lin-7 (Pals1) 21:97 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 0 27 theme Mammalian 0:8 arg1 Crumbs3 10:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 10 28 theme protein 1489:1495 arg1 family 1497:1502 the Crumbs protein family 1478:1502 the Crumbs protein family 1478:1502 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 10 29 theme specialized 1455:1465 arg1 isoform 1467:1473 a specialized isoform 1453:1473 a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction 1453:1587 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 10 29 theme specialized 1455:1465 arg1 CRB3 1445:1448 CRB3 1445:1448 CRB3 1445:1448 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 7 30 theme Drosophila 959:968 arg1 Crumbs 970:975 Drosophila Crumbs 959:975 Drosophila Crumbs 959:975 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 8 31 theme conserved 1248:1256 arg1 site 1274:1277 a conserved N-glycosylation site 1246:1277 a conserved N-glycosylation site 1246:1277 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 4 32 theme retinitis 561:569 arg1 disorders 550:558 the eye disorders 542:558 the eye disorders 542:558 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 4 32 theme retinitis 561:569 arg1 pigmentosa 571:580 retinitis pigmentosa 561:580 retinitis pigmentosa 561:580 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 9 33 theme glycosylation 1367:1379 arg1 site 1381:1384 the conserved N linked glycosylation site 1344:1384 the conserved N linked glycosylation site 1344:1384 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 9 33 theme glycosylation 1367:1379 arg1 necessary 1408:1416 necessary 1408:1416 necessary 1408:1416 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 0 34 attach linked 51:56 arg2 protein 43:49 a small transmembrane protein 21:49 a small transmembrane protein linked to protein associated with Lin-7 (Pals1) 21:97 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 0 34 attach linked 51:56 arg1 protein 61:67 protein 61:67 protein associated with Lin-7 (Pals1) 61:97 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 0 34 attach linked 51:56 arg2 Crumbs3 10:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 8 35 theme divergent 1203:1211 arg1 domains 1227:1233 divergent extracellular domains 1203:1233 divergent extracellular domains 1203:1233 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 2 36 from Overexpression 233:246 arg1 epithelia 272:280 Drosophila epithelia 261:280 Drosophila epithelia 261:280 Overexpression of Crumbs in Drosophila epithelia expands the apical surface and leads to disruption of cell polarity. 12527193 8 37 theme Mouse 1141:1145 arg1 CRB3 1157:1160 Mouse and human CRB3 1141:1160 Mouse and human CRB3 1141:1160 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 4 38 theme Recent 440:445 arg1 work 447:450 Recent work 440:450 Recent work 440:450 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 2 39 theme apical 294:299 arg1 surface 301:307 the apical surface 290:307 the apical surface 290:307 Overexpression of Crumbs in Drosophila epithelia expands the apical surface and leads to disruption of cell polarity. 12527193 1 40 theme epithelial 177:186 arg1 polarity 193:200 epithelial cell polarity 177:200 epithelial cell polarity 177:200 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 0 41 theme transmembrane 29:41 arg1 protein 43:49 a small transmembrane protein 21:49 a small transmembrane protein linked to protein associated with Lin-7 (Pals1) 21:97 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 0 41 theme transmembrane 29:41 arg1 Crumbs3 10:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 6 42 theme length 879:884 arg1 cDNA 886:889 a full length cDNA 872:889 a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3) 872:941 In the current report we have cloned a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3). 12527193 5 43 theme associated 787:796 arg1 PATJ 814:817 PATJ 814:817 PATJ 814:817 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 5 43 theme associated 787:796 arg1 junction 804:811 Pals1 associated tight junction 781:811 Pals1 associated tight junction (PATJ) 781:818 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 10 44 theme apical 1551:1556 arg1 membrane 1558:1565 the apical membrane 1547:1565 the apical membrane 1547:1565 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 1 45 theme cell 188:191 arg1 polarity 193:200 epithelial cell polarity 177:200 epithelial cell polarity 177:200 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 0 46 theme small 23:27 arg1 protein 43:49 a small transmembrane protein 21:49 a small transmembrane protein linked to protein associated with Lin-7 (Pals1) 21:97 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 0 46 theme small 23:27 arg1 Crumbs3 10:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 0:16 Mammalian Crumbs3 is a small transmembrane protein linked to protein associated with Lin-7 (Pals1). 12527193 6 47 theme full 874:877 arg1 cDNA 886:889 a full length cDNA 872:889 a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3) 872:941 In the current report we have cloned a full length cDNA for a human paralogue of CRB1 called Crumbs3 (CRB3). 12527193 5 48 theme mammalian 680:688 arg1 orthologues 690:700 mammalian orthologues 680:700 mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively 680:832 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 8 49 theme human 1151:1155 arg1 CRB3 1157:1160 Mouse and human CRB3 1141:1160 Mouse and human CRB3 1141:1160 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 5 50 theme tight 798:802 arg1 PATJ 814:817 PATJ 814:817 PATJ 814:817 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 5 50 theme tight 798:802 arg1 junction 804:811 Pals1 associated tight junction 781:811 Pals1 associated tight junction (PATJ) 781:818 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 3 51 theme other 393:397 arg1 genes 408:412 two other polarity genes 389:412 two other polarity genes 389:412 Drosophila Crumbs also interacts with two other polarity genes, Stardust and Discs Lost. 12527193 8 52 gly N-glycosylation 1258:1272 arg2 site 1274:1277 a conserved N-glycosylation site 1246:1277 a conserved N-glycosylation site 1246:1277 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 5 53 theme human 645:649 arg1 CRB1 651:654 human CRB1 645:654 human CRB1 645:654 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 9 54 theme linked 1360:1365 arg1 site 1381:1384 the conserved N linked glycosylation site 1344:1384 the conserved N linked glycosylation site 1344:1384 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 9 54 theme linked 1360:1365 arg1 necessary 1408:1416 necessary 1408:1416 necessary 1408:1416 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 2 55 theme cell 336:339 arg1 polarity 341:348 cell polarity 336:348 cell polarity 336:348 Overexpression of Crumbs in Drosophila epithelia expands the apical surface and leads to disruption of cell polarity. 12527193 3 56 theme polarity 399:406 arg1 genes 408:412 two other polarity genes 389:412 two other polarity genes 389:412 Drosophila Crumbs also interacts with two other polarity genes, Stardust and Discs Lost. 12527193 1 57 theme Drosophila 100:109 arg1 Crumbs 111:116 Drosophila Crumbs 100:116 Drosophila Crumbs 100:116 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 1 57 theme Drosophila 100:109 arg1 protein 137:143 a transmembrane protein 121:143 a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development 121:230 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 7 58 contain has 1057:1059 arg1 it 1054:1055 it 1054:1055 it 1054:1055 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 7 58 contain has 1057:1059 arg2 domain 1087:1092 a conserved intracellular domain 1061:1092 a conserved intracellular domain that allows it to complex with Pals1 and PATJ 1061:1138 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 9 59 dep linked 1360:1365 arg1 N 1358:1358 N 1358:1358 N 1358:1358 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 8 60 theme intracellular 1177:1189 arg1 domains 1191:1197 identical intracellular domains 1167:1197 identical intracellular domains 1167:1197 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 8 61 contain have 1162:1165 arg2 domains 1227:1233 divergent extracellular domains 1203:1233 divergent extracellular domains 1203:1233 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 8 61 contain have 1162:1165 arg1 CRB3 1157:1160 Mouse and human CRB3 1141:1160 Mouse and human CRB3 1141:1160 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 8 61 contain have 1162:1165 arg2 domains 1191:1197 identical intracellular domains 1167:1197 identical intracellular domains 1167:1197 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 9 62 gly glycosylation 1367:1379 arg2 site 1381:1384 the conserved N linked glycosylation site 1344:1384 the conserved N linked glycosylation site 1344:1384 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 9 62 gly glycosylation 1367:1379 arg2 necessary 1408:1416 necessary 1408:1416 necessary 1408:1416 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 2 63 theme Crumbs 251:256 arg1 Overexpression 233:246 Overexpression 233:246 Overexpression of Crumbs in Drosophila epithelia 233:280 Overexpression of Crumbs in Drosophila epithelia expands the apical surface and leads to disruption of cell polarity. 12527193 8 64 theme identical 1167:1175 arg1 domains 1191:1197 identical intracellular domains 1167:1197 identical intracellular domains 1167:1197 Mouse and human CRB3 have identical intracellular domains but divergent extracellular domains except for a conserved N-glycosylation site. 12527193 7 65 contain has 992:994 arg1 CRB3 987:990 CRB3 987:990 CRB3 987:990 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 7 65 contain has 992:994 arg2 domain 1023:1028 a very short extracellular domain 996:1028 a very short extracellular domain 996:1028 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 1 66 theme photoreceptor 206:218 arg1 development 220:230 photoreceptor development 206:230 photoreceptor development 206:230 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 5 67 theme Discs 718:722 arg1 orthologues 690:700 mammalian orthologues 680:700 mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively 680:832 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 5 68 with complex 667:673 arg1 orthologues 690:700 mammalian orthologues 680:700 mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively 680:832 Our work has demonstrated that human CRB1 can form a complex with mammalian orthologues of Stardust and Discs Lost, known as protein associated with Lin-7 (Pals1) and Pals1 associated tight junction (PATJ), respectively. 12527193 9 69 theme conserved 1348:1356 arg1 site 1381:1384 the conserved N linked glycosylation site 1344:1384 the conserved N linked glycosylation site 1344:1384 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 9 69 theme conserved 1348:1356 arg1 necessary 1408:1416 necessary 1408:1416 necessary 1408:1416 CRB3 is localized to the apical surface and tight junctions but the conserved N linked glycosylation site does not appear to be necessary for CRB3 apical targeting. 12527193 4 70 theme Leber 586:590 arg1 disorders 550:558 the eye disorders 542:558 the eye disorders 542:558 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 4 70 theme Leber 586:590 arg1 amaurosis 603:611 Leber congenital amaurosis 586:611 Leber congenital amaurosis 586:611 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 10 71 theme family 1497:1502 arg1 isoform 1467:1473 a specialized isoform 1453:1473 a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction 1453:1587 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 10 71 theme family 1497:1502 arg1 CRB3 1445:1448 CRB3 1445:1448 CRB3 1445:1448 CRB3 is a specialized isoform of the Crumbs protein family that is expressed in epithelia and can tie the apical membrane to the tight junction. 12527193 4 72 theme Drosophila 489:498 arg1 Crumbs 500:505 Drosophila Crumbs 489:505 Drosophila Crumbs 489:505 Recent work has identified a human orthologue of Drosophila Crumbs, known as CRB1, that is mutated in the eye disorders, retinitis pigmentosa and Leber congenital amaurosis. 12527193 7 73 with complex 1112:1118 arg1 PATJ 1135:1138 PATJ 1135:1138 PATJ 1135:1138 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 7 73 with complex 1112:1118 arg1 Pals1 1125:1129 Pals1 1125:1129 Pals1 1125:1129 In contrast to Drosophila Crumbs and CRB1, CRB3 has a very short extracellular domain but like these proteins it has a conserved intracellular domain that allows it to complex with Pals1 and PATJ. 12527193 1 74 theme transmembrane 123:135 arg1 Crumbs 111:116 Drosophila Crumbs 100:116 Drosophila Crumbs 100:116 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 12527193 1 74 theme transmembrane 123:135 arg1 protein 137:143 a transmembrane protein 121:143 a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development 121:230 Drosophila Crumbs is a transmembrane protein that plays an important role in epithelial cell polarity and photoreceptor development. 22511793 6 0 with protein 1125:1131 arg1 domain 1168:1173 a highly glycosylated luminal domain 1138:1173 a highly glycosylated luminal domain 1138:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 10 1 theme role 1741:1744 arg1 understanding 1707:1719 the understanding 1703:1719 the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder 1703:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 1 2 theme lobar 252:256 arg1 degeneration 258:269 frontotemporal lobar degeneration 237:269 frontotemporal lobar degeneration 237:269 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 10 3 from understanding 1707:1719 arg1 disorder 1798:1805 an incurable neurodegenerative disorder 1767:1805 an incurable neurodegenerative disorder 1767:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 10 3 from understanding 1707:1719 arg1 FTLD 1761:1764 FTLD 1761:1764 FTLD 1761:1764 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 3 4 theme TMEM106B 545:552 arg1 expression 554:563 TMEM106B expression 545:563 TMEM106B expression 545:563 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 2 5 theme TMEM106B 363:370 arg1 polymorphisms 390:402 TMEM106B single nucleotide polymorphisms 363:402 TMEM106B single nucleotide polymorphisms 363:402 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 10 6 from role 1741:1744 arg1 disorder 1798:1805 an incurable neurodegenerative disorder 1767:1805 an incurable neurodegenerative disorder 1767:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 10 6 from role 1741:1744 arg1 FTLD 1761:1764 FTLD 1761:1764 FTLD 1761:1764 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 1 7 theme major 180:184 arg1 factor 191:196 a major risk factor 178:196 a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology 178:324 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 7 theme major 180:184 arg1 FTLD 272:275 FTLD 272:275 FTLD 272:275 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 7 theme major 180:184 arg1 TMEM106B 151:158 TMEM106B 151:158 TMEM106B 151:158 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 5 8 theme subcellular 904:914 arg1 localization 916:927 subcellular localization 904:927 subcellular localization 904:927 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 6 9 theme luminal 1160:1166 arg1 domain 1168:1173 a highly glycosylated luminal domain 1138:1173 a highly glycosylated luminal domain 1138:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 10 10 theme neurodegenerative 1780:1796 arg1 disorder 1798:1805 an incurable neurodegenerative disorder 1767:1805 an incurable neurodegenerative disorder 1767:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 10 10 theme neurodegenerative 1780:1796 arg1 FTLD 1761:1764 FTLD 1761:1764 FTLD 1761:1764 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 9 11 theme -ATPases 1440:1447 arg1 inhibition 1413:1422 the inhibition 1409:1422 the inhibition of vacuolar H(+)-ATPases 1409:1447 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 3 12 theme patient 583:589 arg1 serum 591:595 patient serum 583:595 patient serum 583:595 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 4 13 theme GRN 696:698 arg1 levels 700:705 GRN levels 696:705 GRN levels 696:705 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 9 14 theme unexpected 1527:1536 arg1 link 1550:1553 an unexpected biochemical link 1524:1553 an unexpected biochemical link to GRN 1524:1560 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 3 15 from levels 573:578 arg1 serum 591:595 patient serum 583:595 patient serum 583:595 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 7 16 theme endoplasmic 1253:1263 arg1 reticulum 1265:1273 the endoplasmic reticulum 1249:1273 the endoplasmic reticulum to late cellular compartments 1249:1303 Glycosylation is partially required for the transport of TMEM106B beyond the endoplasmic reticulum to late cellular compartments. 22511793 6 17 theme sequential 1013:1022 arg1 mutagenesis 1024:1034 sequential mutagenesis 1013:1034 sequential mutagenesis 1013:1034 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 4 18 from expression 735:744 arg1 cells 774:778 heterologous cells 761:778 heterologous cells 761:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 5 19 theme protein 964:970 arg1 orientation 888:898 membrane orientation 879:898 membrane orientation 879:898 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 5 19 theme protein 964:970 arg1 localization 916:927 subcellular localization 904:927 subcellular localization 904:927 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 6 20 theme differential 976:987 arg1 extraction 998:1007 differential membrane extraction 976:1007 differential membrane extraction 976:1007 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 4 21 theme TMEM106B 749:756 arg1 expression 735:744 knockdown or exogenous expression 712:744 knockdown or exogenous expression of TMEM106B in heterologous cells 712:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 2 22 theme nucleotide 379:388 arg1 polymorphisms 390:402 TMEM106B single nucleotide polymorphisms 363:402 TMEM106B single nucleotide polymorphisms 363:402 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 0 23 theme major 94:98 arg1 factor 105:110 a major risk factor 92:110 a major risk factor for frontotemporal lobar degeneration 92:148 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 23 theme major 94:98 arg1 orientation 9:19 Membrane orientation 0:19 Membrane orientation 0:19 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 10 24 theme biochemical 1661:1671 arg1 basis 1693:1697 a biochemical and cell biological basis 1659:1697 a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder 1659:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 0 25 theme Membrane 0:7 arg1 factor 105:110 a major risk factor 92:110 a major risk factor for frontotemporal lobar degeneration 92:148 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 25 theme Membrane 0:7 arg1 orientation 9:19 Membrane orientation 0:19 Membrane orientation 0:19 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 2 26 theme significant 336:346 arg1 association 348:358 The most significant association 327:358 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP 327:424 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 6 27 gly N-glycosylation 1049:1063 arg2 sites 1065:1069 potential N-glycosylation sites 1039:1069 potential N-glycosylation sites 1039:1069 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 1 28 theme DNA-binding 287:297 arg1 protein 299:305 TAR DNA-binding protein 283:305 TAR DNA-binding protein (TDP) 283:311 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 28 theme DNA-binding 287:297 arg1 TDP 308:310 TDP 308:310 TDP 308:310 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 29 with factor 191:196 arg1 protein 299:305 TAR DNA-binding protein 283:305 TAR DNA-binding protein (TDP) 283:311 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 29 with factor 191:196 arg1 TDP 308:310 TDP 308:310 TDP 308:310 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 6 30 theme N-glycosylation 1049:1063 arg1 sites 1065:1069 potential N-glycosylation sites 1039:1069 potential N-glycosylation sites 1039:1069 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 0 31 theme 106B 75:78 arg1 factor 105:110 a major risk factor 92:110 a major risk factor for frontotemporal lobar degeneration 92:148 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 31 theme 106B 75:78 arg1 orientation 9:19 Membrane orientation 0:19 Membrane orientation 0:19 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 31 theme 106B 75:78 arg1 localization 37:48 subcellular localization 25:48 subcellular localization 25:48 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 10 32 theme pathological 1728:1739 arg1 role 1741:1744 the pathological role 1724:1744 the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder 1724:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 0 33 theme transmembrane 53:65 arg1 TMEM106B 81:88 TMEM106B 81:88 TMEM106B 81:88 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 33 theme transmembrane 53:65 arg1 106B 75:78 transmembrane protein 106B 53:78 transmembrane protein 106B (TMEM106B) 53:89 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 4 34 theme heterologous 761:772 arg1 cells 774:778 heterologous cells 761:778 heterologous cells 761:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 9 35 theme vacuolar 1427:1434 arg1 H 1436:1436 vacuolar H 1427:1436 vacuolar H 1427:1436 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 2 36 with association 348:358 arg1 risk 409:412 risk 409:412 risk of FTLD-TDP 409:424 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 10 37 theme biological 1682:1691 arg1 basis 1693:1697 a biochemical and cell biological basis 1659:1697 a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder 1659:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 6 38 theme integral 1107:1114 arg1 TMEM106B 1086:1093 TMEM106B 1086:1093 TMEM106B 1086:1093 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 6 38 theme integral 1107:1114 arg1 protein 1125:1131 a type 2 integral membrane protein 1098:1131 a type 2 integral membrane protein with a highly glycosylated luminal domain 1098:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 3 39 theme Subsequent 485:494 arg1 studies 496:502 Subsequent studies 485:502 Subsequent studies 485:502 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 1 40 theme frontotemporal 237:250 arg1 degeneration 258:269 frontotemporal lobar degeneration 237:269 frontotemporal lobar degeneration 237:269 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 6 41 theme type 1100:1103 arg1 TMEM106B 1086:1093 TMEM106B 1086:1093 TMEM106B 1086:1093 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 6 41 theme type 1100:1103 arg1 protein 1125:1131 a type 2 integral membrane protein 1098:1131 a type 2 integral membrane protein with a highly glycosylated luminal domain 1098:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 10 42 theme incurable 1770:1778 arg1 disorder 1798:1805 an incurable neurodegenerative disorder 1767:1805 an incurable neurodegenerative disorder 1767:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 10 42 theme incurable 1770:1778 arg1 FTLD 1761:1764 FTLD 1761:1764 FTLD 1761:1764 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 1 43 theme association 215:225 arg1 study 227:231 a genome-wide association study 201:231 a genome-wide association study 201:231 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 3 44 theme inverse 517:523 arg1 correlation 525:535 an inverse correlation 514:535 an inverse correlation between TMEM106B expression and GRN levels in patient serum 514:595 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 2 45 located observed 430:437 arg2 association 348:358 The most significant association 327:358 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP 327:424 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 2 45 located observed 430:437 arg1 patients 442:449 patients 442:449 patients with progranulin (GRN) mutations 442:482 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 3 46 theme GRN 569:571 arg1 levels 573:578 GRN levels 569:578 GRN levels 569:578 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 6 47 gly glycosylated 1147:1158 arg1 domain 1168:1173 a highly glycosylated luminal domain 1138:1173 a highly glycosylated luminal domain 1138:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 1 48 theme risk 186:189 arg1 factor 191:196 a major risk factor 178:196 a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology 178:324 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 48 theme risk 186:189 arg1 FTLD 272:275 FTLD 272:275 FTLD 272:275 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 48 theme risk 186:189 arg1 TMEM106B 151:158 TMEM106B 151:158 TMEM106B 151:158 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 6 49 theme glycosylated 1147:1158 arg1 domain 1168:1173 a highly glycosylated luminal domain 1138:1173 a highly glycosylated luminal domain 1138:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 0 50 theme lobar 131:135 arg1 degeneration 137:148 frontotemporal lobar degeneration 116:148 frontotemporal lobar degeneration 116:148 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 9 51 theme TMEM106B 1487:1494 arg1 levels 1477:1482 the levels 1473:1482 the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions 1473:1635 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 5 52 theme TMEM106B 818:825 arg1 function 827:834 TMEM106B function 818:834 TMEM106B function in health and disease 818:856 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 8 53 theme Endogenous 1306:1315 arg1 TMEM106B 1342:1349 Endogenous as well as overexpressed TMEM106B 1306:1349 Endogenous as well as overexpressed TMEM106B 1306:1349 Endogenous as well as overexpressed TMEM106B localizes to late endosomes and lysosomes. 22511793 5 54 from function 827:834 arg1 health 839:844 health 839:844 health 839:844 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 5 54 from function 827:834 arg1 disease 850:856 disease 850:856 disease 850:856 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 5 55 theme membrane 879:886 arg1 orientation 888:898 membrane orientation 879:898 membrane orientation 879:898 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 4 56 theme significant 670:680 arg1 alteration 682:691 a significant alteration 668:691 a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells 668:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 3 57 from expression 554:563 arg1 serum 591:595 patient serum 583:595 patient serum 583:595 Subsequent studies suggested an inverse correlation between TMEM106B expression and GRN levels in patient serum. 22511793 1 58 from factor 191:196 arg1 study 227:231 a genome-wide association study 201:231 a genome-wide association study 201:231 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 10 59 from FTLD 1761:1764 arg1 understanding 1707:1719 the understanding 1703:1719 the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder 1703:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 22511793 6 60 theme potential 1039:1047 arg1 sites 1065:1069 potential N-glycosylation sites 1039:1069 potential N-glycosylation sites 1039:1069 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 8 61 theme overexpressed 1328:1340 arg1 TMEM106B 1342:1349 Endogenous as well as overexpressed TMEM106B 1306:1349 Endogenous as well as overexpressed TMEM106B 1306:1349 Endogenous as well as overexpressed TMEM106B localizes to late endosomes and lysosomes. 22511793 2 62 with patients 442:449 arg1 mutations 474:482 progranulin (GRN) mutations 456:482 progranulin (GRN) mutations 456:482 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 9 63 theme biochemical 1538:1548 arg1 link 1550:1553 an unexpected biochemical link 1524:1553 an unexpected biochemical link to GRN 1524:1560 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 1 64 theme genome-wide 203:213 arg1 association 215:225 a genome-wide association 201:225 a genome-wide association study 201:231 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 7 65 theme TMEM106B 1233:1240 arg1 transport 1220:1228 the transport 1216:1228 the transport of TMEM106B beyond the endoplasmic reticulum to late cellular compartments 1216:1303 Glycosylation is partially required for the transport of TMEM106B beyond the endoplasmic reticulum to late cellular compartments. 22511793 8 66 theme late 1364:1367 arg1 endosomes 1369:1377 late endosomes 1364:1377 late endosomes 1364:1377 Endogenous as well as overexpressed TMEM106B localizes to late endosomes and lysosomes. 22511793 5 67 theme uncharacterized 948:962 arg1 protein 964:970 this completely uncharacterized protein 932:970 this completely uncharacterized protein 932:970 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 6 68 theme membrane 989:996 arg1 extraction 998:1007 differential membrane extraction 976:1007 differential membrane extraction 976:1007 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 4 69 theme exogenous 725:733 arg1 expression 735:744 knockdown or exogenous expression 712:744 knockdown or exogenous expression of TMEM106B in heterologous cells 712:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 0 70 theme risk 100:103 arg1 factor 105:110 a major risk factor 92:110 a major risk factor for frontotemporal lobar degeneration 92:148 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 70 theme risk 100:103 arg1 orientation 9:19 Membrane orientation 0:19 Membrane orientation 0:19 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 5 71 dep orientation 888:898 arg1 the 875:877 the 875:877 the 875:877 To provide a basis for understanding TMEM106B function in health and disease, we investigated the membrane orientation and subcellular localization of this completely uncharacterized protein. 22511793 2 72 theme single 372:377 arg1 polymorphisms 390:402 TMEM106B single nucleotide polymorphisms 363:402 TMEM106B single nucleotide polymorphisms 363:402 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 4 73 theme levels 700:705 arg1 alteration 682:691 a significant alteration 668:691 a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells 668:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 0 74 theme subcellular 25:35 arg1 localization 37:48 subcellular localization 25:48 subcellular localization 25:48 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 2 75 theme FTLD-TDP 417:424 arg1 risk 409:412 risk 409:412 risk of FTLD-TDP 409:424 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 4 76 theme knockdown 712:720 arg1 expression 735:744 knockdown or exogenous expression 712:744 knockdown or exogenous expression of TMEM106B in heterologous cells 712:778 However, in this study, this was not confirmed as we failed to detect a significant alteration of GRN levels upon knockdown or exogenous expression of TMEM106B in heterologous cells. 22511793 6 77 theme membrane 1116:1123 arg1 TMEM106B 1086:1093 TMEM106B 1086:1093 TMEM106B 1086:1093 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 6 77 theme membrane 1116:1123 arg1 protein 1125:1131 a type 2 integral membrane protein 1098:1131 a type 2 integral membrane protein with a highly glycosylated luminal domain 1098:1173 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 1 78 theme TAR 283:285 arg1 protein 299:305 TAR DNA-binding protein 283:305 TAR DNA-binding protein (TDP) 283:311 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 1 78 theme TAR 283:285 arg1 TDP 308:310 TDP 308:310 TDP 308:310 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 6 79 theme sites 1065:1069 arg1 extraction 998:1007 differential membrane extraction 976:1007 differential membrane extraction 976:1007 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 6 79 theme sites 1065:1069 arg1 mutagenesis 1024:1034 sequential mutagenesis 1013:1034 sequential mutagenesis 1013:1034 By differential membrane extraction and sequential mutagenesis of potential N-glycosylation sites, we identified TMEM106B as a type 2 integral membrane protein with a highly glycosylated luminal domain. 22511793 2 80 theme progranulin 456:466 arg1 mutations 474:482 progranulin (GRN) mutations 456:482 progranulin (GRN) mutations 456:482 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 2 81 theme polymorphisms 390:402 arg1 association 348:358 The most significant association 327:358 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP 327:424 The most significant association of TMEM106B single nucleotide polymorphisms with risk of FTLD-TDP was observed in patients with progranulin (GRN) mutations. 22511793 0 82 theme protein 67:73 arg1 TMEM106B 81:88 TMEM106B 81:88 TMEM106B 81:88 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 0 82 theme protein 67:73 arg1 106B 75:78 transmembrane protein 106B 53:78 transmembrane protein 106B (TMEM106B) 53:89 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 9 83 theme same 1621:1624 arg1 conditions 1626:1635 the same conditions 1617:1635 the same conditions 1617:1635 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 0 84 theme frontotemporal 116:129 arg1 degeneration 137:148 frontotemporal lobar degeneration 116:148 frontotemporal lobar degeneration 116:148 Membrane orientation and subcellular localization of transmembrane protein 106B (TMEM106B), a major risk factor for frontotemporal lobar degeneration. 22511793 7 85 theme cellular 1283:1290 arg1 compartments 1292:1303 late cellular compartments 1278:1303 late cellular compartments 1278:1303 Glycosylation is partially required for the transport of TMEM106B beyond the endoplasmic reticulum to late cellular compartments. 22511793 9 86 theme H 1436:1436 arg1 -ATPases 1440:1447 vacuolar H(+)-ATPases 1427:1447 vacuolar H(+)-ATPases 1427:1447 Interestingly, the inhibition of vacuolar H(+)-ATPases significantly increased the levels of TMEM106B, a finding that may provide an unexpected biochemical link to GRN, because this protein is also strongly increased under the same conditions. 22511793 7 87 theme late 1278:1281 arg1 compartments 1292:1303 late cellular compartments 1278:1303 late cellular compartments 1278:1303 Glycosylation is partially required for the transport of TMEM106B beyond the endoplasmic reticulum to late cellular compartments. 22511793 1 88 dep factor 191:196 arg1 pathology 316:324 -43 pathology 312:324 a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology 178:324 TMEM106B was identified as a major risk factor in a genome-wide association study for frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP)-43 pathology. 22511793 10 89 theme TMEM106B 1749:1756 arg1 role 1741:1744 the pathological role 1724:1744 the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder 1724:1805 Our findings provide a biochemical and cell biological basis for the understanding of the pathological role of TMEM106B in FTLD, an incurable neurodegenerative disorder. 14699159 4 0 theme de 952:953 arg1 glycosylation 968:980 de novo partial glycosylation 952:980 de novo partial glycosylation of newly synthesized p90ATF6 952:1009 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 3 1 theme S1P/S2P 644:650 arg1 system 661:666 the S1P/S2P protease system 640:666 the S1P/S2P protease system 640:666 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 8 2 theme multiple 1550:1557 arg1 sites 1575:1579 single or multiple N-glycosylation sites 1540:1579 single or multiple N-glycosylation sites 1540:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 1 3 theme cellular 226:233 arg1 homeostasis 235:245 cellular homeostasis 226:245 cellular homeostasis 226:245 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 7 4 theme transport 1346:1354 arg1 rate 1325:1328 a faster rate 1316:1328 a faster rate of constitutive transport to the Golgi 1316:1367 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 4 5 theme Ca 840:841 arg1 mechanism 878:886 a triggering mechanism 865:886 a triggering mechanism for the UPR 865:898 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 4 5 theme Ca 840:841 arg1 stress 857:862 ER Ca(2+) depletion stress 837:862 ER Ca(2+) depletion stress 837:862 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 4 6 theme partial 960:966 arg1 glycosylation 968:980 de novo partial glycosylation 952:980 de novo partial glycosylation of newly synthesized p90ATF6 952:1009 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 8 7 theme Additional 1491:1500 arg1 analysis 1502:1509 Additional analysis 1491:1509 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites 1491:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 3 8 theme nuclear 682:688 arg1 p60ATF6 695:701 a nuclear form p60ATF6 680:701 a nuclear form p60ATF6 that acts as a transcriptional activator 680:742 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 3 8 theme nuclear 682:688 arg1 activator 734:742 a transcriptional activator 716:742 a transcriptional activator 716:742 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 7 9 from level 1392:1396 arg1 nucleus 1416:1422 the nucleus 1412:1422 the nucleus 1412:1422 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 9 10 theme novel 1777:1781 arg1 mechanism 1783:1791 a novel mechanism 1775:1791 a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR 1775:1925 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 2 11 contain contains 413:420 arg1 protein 404:410 a 90-kDa ER transmembrane protein 378:410 a 90-kDa ER transmembrane protein 378:410 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 11 contain contains 413:420 arg1 ATF6 372:375 ATF6 372:375 ATF6 372:375 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 11 contain contains 413:420 arg2 sites 476:480 three evolutionarily conserved N-linked glycosylation sites 422:480 three evolutionarily conserved N-linked glycosylation sites 422:480 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 1 12 theme protein 161:167 arg1 response 169:176 the unfolded protein response 148:176 the unfolded protein response (UPR) 148:182 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 12 theme protein 161:167 arg1 UPR 179:181 UPR 179:181 UPR 179:181 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 7 13 theme faster 1318:1323 arg1 rate 1325:1328 a faster rate 1316:1328 a faster rate of constitutive transport to the Golgi 1316:1367 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 3 14 theme p90ATF6 555:561 arg1 activation 563:572 p90ATF6 activation 555:572 p90ATF6 activation 555:572 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 8 15 theme constitutive 1600:1611 arg1 activity 1629:1636 higher constitutive transactivating activity 1593:1636 higher constitutive transactivating activity 1593:1636 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 8 16 gly N-glycosylation 1559:1573 arg2 sites 1575:1579 single or multiple N-glycosylation sites 1540:1579 single or multiple N-glycosylation sites 1540:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 9 17 theme glycosylation 1805:1817 arg1 sensor 1852:1857 a sensor 1850:1857 a sensor for ER homeostasis 1850:1876 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 9 17 theme glycosylation 1805:1817 arg1 status 1819:1824 the glycosylation status 1801:1824 the glycosylation status of p90ATF6 1801:1835 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 7 18 theme stress 1483:1488 arg1 absence 1469:1475 the absence 1465:1475 the absence of ER stress 1465:1488 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 4 19 theme triggering 867:876 arg1 mechanism 878:886 a triggering mechanism 865:886 a triggering mechanism for the UPR 865:898 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 4 19 theme triggering 867:876 arg1 stress 857:862 ER Ca(2+) depletion stress 837:862 ER Ca(2+) depletion stress 837:862 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 8 20 theme wild 1643:1646 arg1 type 1648:1651 wild type 1643:1651 wild type ATF6 1643:1656 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 4 21 theme 2+ 843:844 arg1 mechanism 878:886 a triggering mechanism 865:886 a triggering mechanism for the UPR 865:898 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 4 21 theme 2+ 843:844 arg1 stress 857:862 ER Ca(2+) depletion stress 837:862 ER Ca(2+) depletion stress 837:862 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 2 22 link N-linked 453:460 arg1 sites 476:480 three evolutionarily conserved N-linked glycosylation sites 422:480 three evolutionarily conserved N-linked glycosylation sites 422:480 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 4 23 theme depletion 847:855 arg1 mechanism 878:886 a triggering mechanism 865:886 a triggering mechanism for the UPR 865:898 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 4 23 theme depletion 847:855 arg1 stress 857:862 ER Ca(2+) depletion stress 837:862 ER Ca(2+) depletion stress 837:862 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 1 24 theme key 115:117 arg1 activator 135:143 a key transcriptional activator 113:143 a key transcriptional activator 113:143 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 24 theme key 115:117 arg1 ATF6 105:108 ATF6 105:108 ATF6 105:108 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 5 25 theme single 1026:1031 arg1 acid 1039:1042 a single amino acid 1024:1042 a single amino acid 1024:1042 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 7 26 theme stronger 1428:1435 arg1 activity 1453:1460 stronger transactivating activity 1428:1460 stronger transactivating activity 1428:1460 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 2 27 theme glycosylation 462:474 arg1 sites 476:480 three evolutionarily conserved N-linked glycosylation sites 422:480 three evolutionarily conserved N-linked glycosylation sites 422:480 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 5 28 gly glycosylation 1064:1076 arg2 site 1078:1081 the N-linked glycosylation site 1051:1081 the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6 1051:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 3 29 theme glycosylation 793:805 arg1 significance 760:771 the functional significance 745:771 the functional significance of p90ATF6 N-linked glycosylation 745:805 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 3 29 theme glycosylation 793:805 arg1 unknown 810:816 unknown 810:816 unknown 810:816 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 1 30 theme stresses 319:326 arg1 stresses 319:326 environmental and physiological stresses 287:326 environmental and physiological stresses that target the endoplasmic reticulum (ER) 287:369 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 30 theme stresses 319:326 arg1 variety 276:282 a variety 274:282 a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER) 274:369 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 3 31 link N-linked 784:791 arg1 glycosylation 793:805 p90ATF6 N-linked glycosylation 776:805 p90ATF6 N-linked glycosylation 776:805 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 3 32 theme p90ATF6 776:782 arg1 glycosylation 793:805 p90ATF6 N-linked glycosylation 776:805 p90ATF6 N-linked glycosylation 776:805 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 0 33 theme unfolded 78:85 arg1 response 95:102 the unfolded protein response 74:102 the unfolded protein response 74:102 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 2 34 theme carboxyl 493:500 arg1 domain 510:515 its carboxyl luminal domain 489:515 its carboxyl luminal domain 489:515 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 3 35 from ER 600:601 arg1 transit 583:589 transit 583:589 transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system 583:666 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 0 36 theme response 95:102 arg1 activation 60:69 activation 60:69 activation of the unfolded protein response 60:102 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 1 37 theme endoplasmic 344:354 arg1 ER 367:368 ER 367:368 ER 367:368 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 37 theme endoplasmic 344:354 arg1 reticulum 356:364 the endoplasmic reticulum 340:364 the endoplasmic reticulum (ER) 340:369 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 4 38 theme ATF6 926:929 arg1 formation 913:921 the formation 909:921 the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6 909:1009 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 9 39 theme underglycosylated 1683:1699 arg1 proteins 1701:1708 underglycosylated proteins 1683:1708 underglycosylated proteins 1683:1708 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 5 40 theme closest 1083:1089 arg1 site 1078:1081 the N-linked glycosylation site 1051:1081 the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6 1051:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 2 41 theme transmembrane 390:402 arg1 protein 404:410 a 90-kDa ER transmembrane protein 378:410 a 90-kDa ER transmembrane protein 378:410 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 41 theme transmembrane 390:402 arg1 ATF6 372:375 ATF6 372:375 ATF6 372:375 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 42 theme 90-kDa 380:385 arg1 protein 404:410 a 90-kDa ER transmembrane protein 378:410 a 90-kDa ER transmembrane protein 378:410 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 42 theme 90-kDa 380:385 arg1 ATF6 372:375 ATF6 372:375 ATF6 372:375 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 0 43 theme ATF6 22:25 arg1 Underglycosylation 0:17 Underglycosylation 0:17 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 0:103 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 9 44 from accumulation 1667:1678 arg1 ER 1717:1718 the ER 1713:1718 the ER 1713:1718 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 2 45 gly glycosylation 462:474 arg2 three 422:426 three 422:426 three 422:426 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 45 gly glycosylation 462:474 arg2 sites 476:480 three evolutionarily conserved N-linked glycosylation sites 422:480 three evolutionarily conserved N-linked glycosylation sites 422:480 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 46 theme conserved 443:451 arg1 sites 476:480 three evolutionarily conserved N-linked glycosylation sites 422:480 three evolutionarily conserved N-linked glycosylation sites 422:480 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 0 47 theme sensing 38:44 arg1 mechanism 46:54 a novel sensing mechanism 30:54 a novel sensing mechanism for activation of the unfolded protein response 30:102 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 5 48 theme glycosylation 1064:1076 arg1 site 1078:1081 the N-linked glycosylation site 1051:1081 the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6 1051:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 7 49 from activity 1453:1460 arg1 nucleus 1416:1422 the nucleus 1412:1422 the nucleus 1412:1422 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 4 50 theme p90ATF6 1003:1009 arg1 glycosylation 968:980 de novo partial glycosylation 952:980 de novo partial glycosylation of newly synthesized p90ATF6 952:1009 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 8 51 theme p90ATF6 1514:1520 arg1 mutants 1522:1528 p90ATF6 mutants 1514:1528 p90ATF6 mutants targeting single or multiple N-glycosylation sites 1514:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 6 52 gly N-glycoprotein 1256:1269 arg1 N-glycoprotein 1256:1269 N-glycoprotein 1256:1269 N-glycoprotein 1256:1269 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 6 53 theme p90ATF6 1180:1186 arg1 association 1188:1198 p90ATF6 association 1180:1198 p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein 1180:1269 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 5 54 theme amino 1033:1037 arg1 acid 1039:1042 a single amino acid 1024:1042 a single amino acid 1024:1042 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 7 55 theme constitutive 1333:1344 arg1 transport 1346:1354 constitutive transport 1333:1354 constitutive transport to the Golgi 1333:1367 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 3 56 theme protease 652:659 arg1 system 661:666 the S1P/S2P protease system 640:666 the S1P/S2P protease system 640:666 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 9 57 theme ER 1863:1864 arg1 homeostasis 1866:1876 ER homeostasis 1863:1876 ER homeostasis 1863:1876 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 5 58 theme p90ATF6 1119:1125 arg1 terminus 1107:1114 the carboxyl terminus 1094:1114 the carboxyl terminus of p90ATF6 1094:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 4 59 theme ER 837:838 arg1 mechanism 878:886 a triggering mechanism 865:886 a triggering mechanism for the UPR 865:898 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 4 59 theme ER 837:838 arg1 stress 857:862 ER Ca(2+) depletion stress 837:862 ER Ca(2+) depletion stress 837:862 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 3 60 theme form 690:693 arg1 p60ATF6 695:701 a nuclear form p60ATF6 680:701 a nuclear form p60ATF6 that acts as a transcriptional activator 680:742 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 3 60 theme form 690:693 arg1 activator 734:742 a transcriptional activator 716:742 a transcriptional activator 716:742 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 8 61 theme N-glycosylation 1559:1573 arg1 sites 1575:1579 single or multiple N-glycosylation sites 1540:1579 single or multiple N-glycosylation sites 1540:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 1 62 theme unfolded 152:159 arg1 response 169:176 the unfolded protein response 148:176 the unfolded protein response (UPR) 148:182 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 62 theme unfolded 152:159 arg1 UPR 179:181 UPR 179:181 UPR 179:181 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 63 theme response 169:176 arg1 activator 135:143 a key transcriptional activator 113:143 a key transcriptional activator 113:143 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 63 theme response 169:176 arg1 ATF6 105:108 ATF6 105:108 ATF6 105:108 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 8 64 theme higher 1593:1598 arg1 activity 1629:1636 higher constitutive transactivating activity 1593:1636 higher constitutive transactivating activity 1593:1636 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 7 65 theme ER 1480:1481 arg1 stress 1483:1488 ER stress 1480:1488 ER stress 1480:1488 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 6 66 theme major 1221:1225 arg1 calreticulin 1205:1216 calreticulin 1205:1216 calreticulin 1205:1216 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 6 66 theme major 1221:1225 arg1 chaperone 1242:1250 a major Ca(2+)-binding chaperone 1219:1250 a major Ca(2+)-binding chaperone for N-glycoprotein 1219:1269 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 8 67 theme transactivating 1613:1627 arg1 activity 1629:1636 higher constitutive transactivating activity 1593:1636 higher constitutive transactivating activity 1593:1636 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 9 68 theme p90ATF6 1829:1835 arg1 sensor 1852:1857 a sensor 1850:1857 a sensor for ER homeostasis 1850:1876 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 9 68 theme p90ATF6 1829:1835 arg1 status 1819:1824 the glycosylation status 1801:1824 the glycosylation status of p90ATF6 1801:1835 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 9 69 gly glycosylation 1805:1817 arg1 p90ATF6 1829:1835 p90ATF6 1829:1835 p90ATF6 1829:1835 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 3 70 theme transcriptional 718:732 arg1 p60ATF6 695:701 a nuclear form p60ATF6 680:701 a nuclear form p60ATF6 that acts as a transcriptional activator 680:742 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 3 70 theme transcriptional 718:732 arg1 activator 734:742 a transcriptional activator 716:742 a transcriptional activator 716:742 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 8 71 theme type 1648:1651 arg1 ATF6 1653:1656 wild type ATF6 1643:1656 wild type ATF6 1643:1656 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 7 72 theme transactivating 1437:1451 arg1 activity 1453:1460 stronger transactivating activity 1428:1460 stronger transactivating activity 1428:1460 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 6 73 with association 1188:1198 arg1 calreticulin 1205:1216 calreticulin 1205:1216 calreticulin 1205:1216 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 6 73 with association 1188:1198 arg1 chaperone 1242:1250 a major Ca(2+)-binding chaperone 1219:1250 a major Ca(2+)-binding chaperone for N-glycoprotein 1219:1269 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 4 74 gly glycosylation 968:980 arg1 p90ATF6 1003:1009 newly synthesized p90ATF6 985:1009 newly synthesized p90ATF6 985:1009 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 1 75 theme transcriptional 119:133 arg1 activator 135:143 a key transcriptional activator 113:143 a key transcriptional activator 113:143 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 1 75 theme transcriptional 119:133 arg1 ATF6 105:108 ATF6 105:108 ATF6 105:108 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 6 76 theme -binding 1233:1240 arg1 calreticulin 1205:1216 calreticulin 1205:1216 calreticulin 1205:1216 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 6 76 theme -binding 1233:1240 arg1 chaperone 1242:1250 a major Ca(2+)-binding chaperone 1219:1250 a major Ca(2+)-binding chaperone for N-glycoprotein 1219:1269 This mutation sharply reduces p90ATF6 association with calreticulin, a major Ca(2+)-binding chaperone for N-glycoprotein. 14699159 1 77 theme physiological 305:317 arg1 stresses 319:326 environmental and physiological stresses 287:326 environmental and physiological stresses that target the endoplasmic reticulum (ER) 287:369 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 2 78 theme N-linked 453:460 arg1 sites 476:480 three evolutionarily conserved N-linked glycosylation sites 422:480 three evolutionarily conserved N-linked glycosylation sites 422:480 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 0 79 theme protein 87:93 arg1 response 95:102 the unfolded protein response 74:102 the unfolded protein response 74:102 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 9 80 gly underglycosylated 1683:1699 arg1 proteins 1701:1708 underglycosylated proteins 1683:1708 underglycosylated proteins 1683:1708 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 7 81 theme p60ATF6 1401:1407 arg1 level 1392:1396 a higher level 1383:1396 a higher level of p60ATF6 in the nucleus 1383:1422 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 7 81 theme p60ATF6 1401:1407 arg1 activity 1453:1460 stronger transactivating activity 1428:1460 stronger transactivating activity 1428:1460 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 2 82 theme luminal 502:508 arg1 domain 510:515 its carboxyl luminal domain 489:515 its carboxyl luminal domain 489:515 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 3 83 theme N-linked 784:791 arg1 glycosylation 793:805 p90ATF6 N-linked glycosylation 776:805 p90ATF6 N-linked glycosylation 776:805 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 4 84 dep de 952:953 arg1 novo 955:958 novo 955:958 novo 955:958 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 9 85 theme ATF6 1892:1895 arg1 activation 1897:1906 ATF6 activation 1892:1906 ATF6 activation 1892:1906 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 9 86 theme proteins 1701:1708 arg1 inducer 1732:1738 a potent inducer 1723:1738 a potent inducer for the UPR 1723:1750 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 9 86 theme proteins 1701:1708 arg1 accumulation 1667:1678 accumulation 1667:1678 accumulation of underglycosylated proteins in the ER 1667:1718 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 2 87 theme ER 387:388 arg1 protein 404:410 a 90-kDa ER transmembrane protein 378:410 a 90-kDa ER transmembrane protein 378:410 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 2 87 theme ER 387:388 arg1 ATF6 372:375 ATF6 372:375 ATF6 372:375 ATF6, a 90-kDa ER transmembrane protein, contains three evolutionarily conserved N-linked glycosylation sites within its carboxyl luminal domain. 14699159 0 88 theme novel 32:36 arg1 mechanism 46:54 a novel sensing mechanism 30:54 a novel sensing mechanism for activation of the unfolded protein response 30:102 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 5 89 theme carboxyl 1098:1105 arg1 terminus 1107:1114 the carboxyl terminus 1094:1114 the carboxyl terminus of p90ATF6 1094:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 0 90 gly Underglycosylation 0:17 arg1 ATF6 22:25 ATF6 22:25 ATF6 22:25 Underglycosylation of ATF6 as a novel sensing mechanism for activation of the unfolded protein response. 14699159 3 91 theme functional 749:758 arg1 significance 760:771 the functional significance 745:771 the functional significance of p90ATF6 N-linked glycosylation 745:805 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 3 91 theme functional 749:758 arg1 unknown 810:816 unknown 810:816 unknown 810:816 Although it is well established that p90ATF6 activation requires transit from the ER to the Golgi, where it is cleaved by the S1P/S2P protease system to generate a nuclear form p60ATF6 that acts as a transcriptional activator, the functional significance of p90ATF6 N-linked glycosylation is unknown. 14699159 5 92 theme N-linked 1055:1062 arg1 site 1078:1081 the N-linked glycosylation site 1051:1081 the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6 1051:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 7 93 theme higher 1385:1390 arg1 level 1392:1396 a higher level 1383:1396 a higher level of p60ATF6 in the nucleus 1383:1422 We further determined that ATF6(f) exhibits a faster rate of constitutive transport to the Golgi, resulting in a higher level of p60ATF6 in the nucleus and stronger transactivating activity in the absence of ER stress. 14699159 9 94 theme potent 1725:1730 arg1 inducer 1732:1738 a potent inducer 1723:1738 a potent inducer for the UPR 1723:1750 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 9 94 theme potent 1725:1730 arg1 accumulation 1667:1678 accumulation 1667:1678 accumulation of underglycosylated proteins in the ER 1667:1718 Because accumulation of underglycosylated proteins in the ER is a potent inducer for the UPR, these studies uncover a novel mechanism whereby the glycosylation status of p90ATF6 can serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the UPR. 14699159 1 95 theme environmental 287:299 arg1 stresses 319:326 environmental and physiological stresses 287:326 environmental and physiological stresses that target the endoplasmic reticulum (ER) 287:369 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 5 96 link N-linked 1055:1062 arg1 site 1078:1081 the N-linked glycosylation site 1051:1081 the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6 1051:1125 By mutating a single amino acid within the N-linked glycosylation site closest to the carboxyl terminus of p90ATF6, we recreated ATF6(f). 14699159 8 97 theme mutants 1522:1528 arg1 analysis 1502:1509 Additional analysis 1491:1509 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites 1491:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 14699159 1 98 theme mammalian 198:206 arg1 cells 208:212 mammalian cells 198:212 mammalian cells 198:212 ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to a variety of environmental and physiological stresses that target the endoplasmic reticulum (ER). 14699159 4 99 theme synthesized 991:1001 arg1 p90ATF6 1003:1009 newly synthesized p90ATF6 985:1009 newly synthesized p90ATF6 985:1009 Here we show that ER Ca(2+) depletion stress, a triggering mechanism for the UPR, induces the formation of ATF6(f), which represents de novo partial glycosylation of newly synthesized p90ATF6. 14699159 8 100 theme single 1540:1545 arg1 sites 1575:1579 single or multiple N-glycosylation sites 1540:1579 single or multiple N-glycosylation sites 1540:1579 Additional analysis of p90ATF6 mutants targeting single or multiple N-glycosylation sites also showed higher constitutive transactivating activity than wild type ATF6. 10769135 7 0 theme Structural 1152:1161 arg1 studies 1163:1169 Structural studies 1152:1169 Structural studies 1152:1169 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 9 1 theme water 1570:1574 arg1 molecule 1576:1583 a water molecule 1568:1583 a water molecule 1568:1583 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 6 2 theme catalytic 1132:1140 arg1 activity 1142:1149 catalytic activity 1132:1149 catalytic activity 1132:1149 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 4 3 theme binding 605:611 arg1 subsites 613:620 five binding subsites 600:620 five binding subsites 600:620 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 10 4 theme chains 1837:1842 arg1 substitution 1812:1823 substitution 1812:1823 substitution of the side chains of either Glu233 or Asp300 1812:1869 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 1 5 theme active 161:166 arg1 region 173:178 the active site region 157:178 the active site region of human pancreatic alpha-amylase 157:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 11 6 theme pancreatic 2004:2013 arg1 alpha-amylase 2015:2027 human pancreatic alpha-amylase 1998:2027 human pancreatic alpha-amylase 1998:2027 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 11 7 with complex 2037:2043 arg1 acarbose 2050:2057 acarbose 2050:2057 acarbose 2050:2057 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 1 8 theme region 173:178 arg1 study 148:152 a multifaceted study 133:152 a multifaceted study of the active site region of human pancreatic alpha-amylase 133:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 6 9 theme residue 1077:1083 arg1 substitution 1056:1067 substitution 1056:1067 substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity 1056:1149 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 10 10 dep decrease 1918:1925 arg1 -fold 1912:1916 -fold 1912:1916 -fold 1912:1916 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 1 11 theme human 183:187 arg1 alpha-amylase 200:212 human pancreatic alpha-amylase 183:212 human pancreatic alpha-amylase 183:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 9 12 theme N-linked 1618:1625 arg1 bond 1638:1641 the inhibitor N-linked glycosidic bond 1604:1641 the inhibitor N-linked glycosidic bond 1604:1641 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 7 13 theme transglycosylation 1311:1328 arg1 reactions 1330:1338 transglycosylation reactions 1311:1338 transglycosylation reactions 1311:1338 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 1 14 theme alpha-amylase 200:212 arg1 region 173:178 the active site region 157:178 the active site region of human pancreatic alpha-amylase 157:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 10 15 dep -fold 1912:1916 arg1 10 1907:1908 10 1907:1908 10 1907:1908 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 7 16 theme hydrolysis 1296:1305 arg1 series 1286:1291 a series 1284:1291 a series of hydrolysis and transglycosylation reactions 1284:1338 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 11 17 theme Asp300Asn 1977:1985 arg1 variant 1987:1993 the Asp300Asn variant 1973:1993 the Asp300Asn variant of human pancreatic alpha-amylase 1973:2027 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 11 18 theme Structural 1950:1959 arg1 analyses 1961:1968 Structural analyses 1950:1968 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase 1950:2027 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 8 19 theme site 1413:1416 arg1 region 1418:1423 the active site region 1402:1423 the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage 1402:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 6 20 dep drop 1124:1127 arg1 -fold 1118:1122 -fold 1118:1122 -fold 1118:1122 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 2 21 theme kinetic 241:247 arg1 analyses 249:256 novel kinetic analyses 235:256 novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides 235:320 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 6 22 theme Further 950:956 arg1 studies 981:987 Further kinetic and structural studies 950:987 Further kinetic and structural studies 950:987 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 4 23 theme active 631:636 arg1 site 638:641 the active site 627:641 the active site 627:641 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 0 24 theme site 61:64 arg1 mapping 8:14 Subsite mapping 0:14 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques 0:120 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 1 25 theme multifaceted 135:146 arg1 study 148:152 a multifaceted study 133:152 a multifaceted study of the active site region of human pancreatic alpha-amylase 133:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 11 26 theme Asp300 2097:2102 arg1 importance 2083:2092 the importance 2079:2092 the importance of Asp300 to the mode of inhibitor binding 2079:2135 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 8 27 theme N-linked 1434:1441 arg1 bond 1456:1459 its N-linked "glycosidic" bond 1430:1459 its N-linked "glycosidic" bond positioned at the normal site of cleavage 1430:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 0 28 theme structural 74:83 arg1 techniques 111:120 structural, kinetic, and mutagenesis techniques 74:120 structural, kinetic, and mutagenesis techniques 74:120 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 11 29 theme binding 2129:2135 arg1 mode 2111:2114 the mode 2107:2114 the mode of inhibitor binding 2107:2135 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 8 30 theme glycosidic 1444:1453 arg1 bond 1456:1459 its N-linked "glycosidic" bond 1430:1459 its N-linked "glycosidic" bond positioned at the normal site of cleavage 1430:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 4 31 theme significant 698:708 arg1 subsite 674:680 an additional glycon subsite 653:680 an additional glycon subsite 653:680 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 4 31 theme significant 698:708 arg1 factor 710:715 a significant factor 696:715 a significant factor 696:715 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 3 32 from sugars 535:540 arg1 sites 569:573 the glycon (-1, -2, -3) sites 545:573 the glycon (-1, -2, -3) sites 545:573 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 0 33 theme kinetic 86:92 arg1 techniques 111:120 structural, kinetic, and mutagenesis techniques 74:120 structural, kinetic, and mutagenesis techniques 74:120 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 5 34 theme groups 889:894 arg1 importance 860:869 the importance 856:869 the importance of these hydroxyl groups for catalysis 856:908 Kinetic parameters for the cleavage of substrates modified at the 2 and 4' ' positions also highlight the importance of these hydroxyl groups for catalysis and identify the rate-determining step. 10769135 4 35 theme additional 656:665 arg1 subsite 674:680 an additional glycon subsite 653:680 an additional glycon subsite 653:680 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 4 35 theme additional 656:665 arg1 factor 710:715 a significant factor 696:715 a significant factor 696:715 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 2 36 theme cleavage 334:341 arg1 pattern 350:356 an overall cleavage action pattern 323:356 an overall cleavage action pattern for this enzyme 323:372 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 11 37 with analyses 1961:1968 arg1 acarbose 2050:2057 acarbose 2050:2057 acarbose 2050:2057 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 0 38 theme Subsite 0:6 arg1 mapping 8:14 Subsite mapping 0:14 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques 0:120 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 8 39 theme normal 1479:1484 arg1 cleavage 1494:1501 cleavage 1494:1501 cleavage 1494:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 8 39 theme normal 1479:1484 arg1 site 1486:1489 the normal site 1475:1489 the normal site of cleavage 1475:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 3 40 theme leaving 474:480 arg1 sites 498:502 aglycone sites +1 and +2 489:512 sites 498:502 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 3 40 theme leaving 474:480 arg1 +2 511:512 +2 511:512 +2 511:512 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 3 40 theme leaving 474:480 arg1 residue 452:458 a maltose residue 442:458 a maltose residue 442:458 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 3 40 theme leaving 474:480 arg1 group 482:486 the leaving group 470:486 the leaving group (aglycone sites +1 and +2) 470:513 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 9 41 theme acid/base 1731:1739 arg1 catalyst 1741:1748 acid/base catalyst 1731:1748 acid/base catalyst 1731:1748 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 0 42 theme human 23:27 arg1 site 61:64 the human pancreatic alpha-amylase active site 19:64 the human pancreatic alpha-amylase active site 19:64 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 4 43 dep appears 587:593 arg1 although 644:651 although 644:651 although 644:651 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 7 44 theme acarbose 1230:1237 arg1 structure 1217:1225 the original pseudo-tetrasaccharide structure 1181:1225 the original pseudo-tetrasaccharide structure of acarbose 1181:1237 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 3 45 theme binding/cleavage 408:423 arg1 mode 425:428 The preferred binding/cleavage mode 394:428 The preferred binding/cleavage mode 394:428 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 0 46 theme alpha-amylase 40:52 arg1 site 61:64 the human pancreatic alpha-amylase active site 19:64 the human pancreatic alpha-amylase active site 19:64 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 9 47 theme Glu233 1538:1543 arg1 chains 1528:1533 the side chains 1519:1533 the side chains of Glu233 and Asp300 1519:1554 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 9 48 theme reaction 1757:1764 arg1 mechanism 1766:1774 the reaction mechanism 1753:1774 the reaction mechanism 1753:1774 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 2 49 theme action 343:348 arg1 pattern 350:356 an overall cleavage action pattern 323:356 an overall cleavage action pattern for this enzyme 323:372 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 9 50 theme Asp300 1549:1554 arg1 chains 1528:1533 the side chains 1519:1533 the side chains of Glu233 and Asp300 1519:1554 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 7 51 theme original 1185:1192 arg1 structure 1217:1225 the original pseudo-tetrasaccharide structure 1181:1225 the original pseudo-tetrasaccharide structure of acarbose 1181:1237 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 10 52 theme catalytic 1930:1938 arg1 activity 1940:1947 catalytic activity 1930:1947 catalytic activity 1930:1947 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 4 53 theme longer 735:740 arg1 substrates 742:751 longer substrates 735:751 longer substrates 735:751 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 5 54 theme rate-determining 927:942 arg1 step 944:947 the rate-determining step 923:947 the rate-determining step 923:947 Kinetic parameters for the cleavage of substrates modified at the 2 and 4' ' positions also highlight the importance of these hydroxyl groups for catalysis and identify the rate-determining step. 10769135 8 55 with region 1418:1423 arg1 bond 1456:1459 its N-linked "glycosidic" bond 1430:1459 its N-linked "glycosidic" bond positioned at the normal site of cleavage 1430:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 11 56 theme human 1998:2002 arg1 alpha-amylase 2015:2027 human pancreatic alpha-amylase 1998:2027 human pancreatic alpha-amylase 1998:2027 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 6 57 theme likely 1018:1023 arg1 nucleophile 1025:1035 the likely nucleophile 1014:1035 the likely nucleophile in catalysis 1014:1048 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 2 58 theme malto-oligosaccharyl 291:310 arg1 fluorides 312:320 malto-oligosaccharyl fluorides 291:320 malto-oligosaccharyl fluorides 291:320 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 3 59 theme glycon 549:554 arg1 sites 569:573 the glycon (-1, -2, -3) sites 545:573 the glycon (-1, -2, -3) sites 545:573 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 9 60 from role 1723:1726 arg1 mechanism 1766:1774 the reaction mechanism 1753:1774 the reaction mechanism 1753:1774 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 11 61 theme variant 1987:1993 arg1 analyses 1961:1968 Structural analyses 1950:1968 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase 1950:2027 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 11 61 theme variant 1987:1993 arg1 complex 2037:2043 its complex 2033:2043 its complex with acarbose 2033:2057 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 5 62 theme substrates 793:802 arg1 cleavage 781:788 the cleavage 777:788 the cleavage of substrates modified at the 2 and 4' ' positions 777:839 Kinetic parameters for the cleavage of substrates modified at the 2 and 4' ' positions also highlight the importance of these hydroxyl groups for catalysis and identify the rate-determining step. 10769135 10 63 theme side 1832:1835 arg1 chains 1837:1842 the side chains 1828:1842 the side chains of either Glu233 or Asp300 1828:1869 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 2 64 theme overall 326:332 arg1 pattern 350:356 an overall cleavage action pattern 323:356 an overall cleavage action pattern for this enzyme 323:372 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 1 65 theme site 168:171 arg1 region 173:178 the active site region 157:178 the active site region of human pancreatic alpha-amylase 157:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 11 66 theme alpha-amylase 2015:2027 arg1 variant 1987:1993 the Asp300Asn variant 1973:1993 the Asp300Asn variant of human pancreatic alpha-amylase 1973:2027 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 10 67 theme Asp300 1864:1869 arg1 chains 1837:1842 the side chains 1828:1842 the side chains of either Glu233 or Asp300 1828:1869 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 9 68 theme inhibitor 1608:1616 arg1 bond 1638:1641 the inhibitor N-linked glycosidic bond 1604:1641 the inhibitor N-linked glycosidic bond 1604:1641 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 9 69 dep along 1557:1561 arg1 with 1563:1566 with 1563:1566 with 1563:1566 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 10 70 theme Glu233 1854:1859 arg1 chains 1837:1842 the side chains 1828:1842 the side chains of either Glu233 or Asp300 1828:1869 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 3 71 dep sites 569:573 arg1 -3 565:566 -3 565:566 -3 565:566 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 9 72 theme glycosidic 1627:1636 arg1 bond 1638:1641 the inhibitor N-linked glycosidic bond 1604:1641 the inhibitor N-linked glycosidic bond 1604:1641 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 8 73 theme active 1406:1411 arg1 region 1418:1423 the active site region 1402:1423 the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage 1402:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 7 74 theme reactions 1330:1338 arg1 series 1286:1291 a series 1284:1291 a series of hydrolysis and transglycosylation reactions 1284:1338 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 2 75 theme analyses 249:256 arg1 series 225:230 a series 223:230 a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides 223:320 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 6 76 theme kinetic 958:964 arg1 studies 981:987 Further kinetic and structural studies 950:987 Further kinetic and structural studies 950:987 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 6 77 dep -fold 1118:1122 arg1 10 1113:1114 10 1113:1114 10 1113:1114 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 2 78 theme novel 235:239 arg1 analyses 249:256 novel kinetic analyses 235:256 novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides 235:320 Through a series of novel kinetic analyses using malto-oligosaccharides and malto-oligosaccharyl fluorides, an overall cleavage action pattern for this enzyme has been developed. 10769135 8 79 theme " 1454:1454 arg1 bond 1456:1459 its N-linked "glycosidic" bond 1430:1459 its N-linked "glycosidic" bond positioned at the normal site of cleavage 1430:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 6 80 from drop 1124:1127 arg1 activity 1142:1149 catalytic activity 1132:1149 catalytic activity 1132:1149 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 4 81 from factor 710:715 arg1 binding 724:730 the binding 720:730 the binding of longer substrates 720:751 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 5 82 theme hydroxyl 880:887 arg1 groups 889:894 these hydroxyl groups 874:894 these hydroxyl groups 874:894 Kinetic parameters for the cleavage of substrates modified at the 2 and 4' ' positions also highlight the importance of these hydroxyl groups for catalysis and identify the rate-determining step. 10769135 8 83 link N-linked 1434:1441 arg1 bond 1456:1459 its N-linked "glycosidic" bond 1430:1459 its N-linked "glycosidic" bond positioned at the normal site of cleavage 1430:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 0 84 theme mutagenesis 99:109 arg1 techniques 111:120 structural, kinetic, and mutagenesis techniques 74:120 structural, kinetic, and mutagenesis techniques 74:120 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 11 85 theme inhibitor 2119:2127 arg1 binding 2129:2135 inhibitor binding 2119:2135 inhibitor binding 2119:2135 Structural analyses of the Asp300Asn variant of human pancreatic alpha-amylase and its complex with acarbose clearly demonstrate the importance of Asp300 to the mode of inhibitor binding. 10769135 3 86 theme preferred 398:406 arg1 mode 425:428 The preferred binding/cleavage mode 394:428 The preferred binding/cleavage mode 394:428 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 3 87 theme maltose 444:450 arg1 residue 452:458 a maltose residue 442:458 a maltose residue 442:458 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 3 87 theme maltose 444:450 arg1 group 482:486 the leaving group 470:486 the leaving group (aglycone sites +1 and +2) 470:513 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 5 88 theme Kinetic 754:760 arg1 parameters 762:771 Kinetic parameters 754:771 Kinetic parameters for the cleavage of substrates modified at the 2 and 4' ' positions 754:839 Kinetic parameters for the cleavage of substrates modified at the 2 and 4' ' positions also highlight the importance of these hydroxyl groups for catalysis and identify the rate-determining step. 10769135 8 89 theme cleavage 1494:1501 arg1 cleavage 1494:1501 cleavage 1494:1501 cleavage 1494:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 8 89 theme cleavage 1494:1501 arg1 site 1486:1489 the normal site 1475:1489 the normal site of cleavage 1475:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 0 90 theme pancreatic 29:38 arg1 site 61:64 the human pancreatic alpha-amylase active site 19:64 the human pancreatic alpha-amylase active site 19:64 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 1 91 theme pancreatic 189:198 arg1 alpha-amylase 200:212 human pancreatic alpha-amylase 183:212 human pancreatic alpha-amylase 183:212 We report a multifaceted study of the active site region of human pancreatic alpha-amylase. 10769135 9 92 theme side 1523:1526 arg1 chains 1528:1533 the side chains 1519:1533 the side chains of Glu233 and Asp300 1519:1554 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 6 93 theme structural 970:979 arg1 studies 981:987 Further kinetic and structural studies 950:987 Further kinetic and structural studies 950:987 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 9 94 theme catalyst 1741:1748 arg1 role 1723:1726 the role 1719:1726 the role of acid/base catalyst in the reaction mechanism 1719:1774 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 4 95 theme substrates 742:751 arg1 binding 724:730 the binding 720:730 the binding of longer substrates 720:751 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 0 96 theme active 54:59 arg1 site 61:64 the human pancreatic alpha-amylase active site 19:64 the human pancreatic alpha-amylase active site 19:64 Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. 10769135 7 97 theme pseudo-tetrasaccharide 1194:1215 arg1 structure 1217:1225 the original pseudo-tetrasaccharide structure 1181:1225 the original pseudo-tetrasaccharide structure of acarbose 1181:1237 Structural studies show that the original pseudo-tetrasaccharide structure of acarbose is modified upon binding, presumably through a series of hydrolysis and transglycosylation reactions. 10769135 8 98 theme end 1345:1347 arg1 result 1349:1354 The end result 1341:1354 The end result 1341:1354 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 8 98 theme end 1345:1347 arg1 moiety 1384:1389 a pseudo-pentasaccharide moiety 1359:1389 a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage 1359:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 4 99 theme glycon 667:672 arg1 subsite 674:680 an additional glycon subsite 653:680 an additional glycon subsite 653:680 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 4 99 theme glycon 667:672 arg1 factor 710:715 a significant factor 696:715 a significant factor 696:715 Overall it appears that five binding subsites span the active site, although an additional glycon subsite appears to be a significant factor in the binding of longer substrates. 10769135 10 100 from decrease 1918:1925 arg1 activity 1940:1947 catalytic activity 1930:1947 catalytic activity 1930:1947 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 10 101 theme kinetic 1785:1791 arg1 analyses 1793:1800 kinetic analyses 1785:1800 kinetic analyses 1785:1800 Indeed, kinetic analyses show that substitution of the side chains of either Glu233 or Asp300 leads to as much as a approximately 10(3)-fold decrease in catalytic activity. 10769135 6 102 from nucleophile 1025:1035 arg1 catalysis 1040:1048 catalysis 1040:1048 catalysis 1040:1048 Further kinetic and structural studies pinpoint Asp197 as being the likely nucleophile in catalysis, with substitution of this residue leading to an approximately 10(6)-fold drop in catalytic activity. 10769135 3 103 theme aglycone 489:496 arg1 sites 498:502 aglycone sites +1 and +2 489:512 sites 498:502 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 3 103 theme aglycone 489:496 arg1 group 482:486 the leaving group 470:486 the leaving group (aglycone sites +1 and +2) 470:513 The preferred binding/cleavage mode occurs when a maltose residue serves as the leaving group (aglycone sites +1 and +2) and there are three sugars in the glycon (-1, -2, -3) sites. 10769135 9 104 link N-linked 1618:1625 arg1 bond 1638:1641 the inhibitor N-linked glycosidic bond 1604:1641 the inhibitor N-linked glycosidic bond 1604:1641 Interestingly, the side chains of Glu233 and Asp300, along with a water molecule, are aligned about the inhibitor N-linked glycosidic bond in a manner suggesting that these might act individually or collectively in the role of acid/base catalyst in the reaction mechanism. 10769135 8 105 theme pseudo-pentasaccharide 1361:1382 arg1 result 1349:1354 The end result 1341:1354 The end result 1341:1354 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 10769135 8 105 theme pseudo-pentasaccharide 1361:1382 arg1 moiety 1384:1389 a pseudo-pentasaccharide moiety 1359:1389 a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage 1359:1501 The end result is a pseudo-pentasaccharide moiety that spans the active site region with its N-linked "glycosidic" bond positioned at the normal site of cleavage. 15961631 1 0 theme immune 123:128 arg1 responses 130:138 immune responses 123:138 immune responses 123:138 Toll-like receptors (TLRs) play key roles in activating immune responses during infection. 15961631 0 1 theme ectodomain 55:64 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain 0:64 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain. 15961631 3 2 theme LRR 340:342 arg1 motif 344:348 the 24-residue LRR motif 325:348 the 24-residue LRR motif 325:348 Asparagines conserved in the 24-residue LRR motif contribute extensive hydrogen-bonding networks for solenoid stabilization. 15961631 2 3 theme leucine-rich 271:282 arg1 LRRs 293:296 LRRs 293:296 LRRs 293:296 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 2 3 theme leucine-rich 271:282 arg1 repeats 284:290 23 leucine-rich repeats 268:290 23 leucine-rich repeats (LRRs) 268:297 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 5 4 theme TLR3-specific 614:626 arg1 insertion 632:640 a TLR3-specific LRR insertion 612:640 a TLR3-specific LRR insertion 612:640 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 5 5 theme second 742:747 arg1 insertion 749:757 a second insertion 740:757 a second insertion 740:757 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 5 6 theme LRR 628:630 arg1 insertion 632:640 a TLR3-specific LRR insertion 612:640 a TLR3-specific LRR insertion 612:640 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 3 7 theme 24-residue 329:338 arg1 motif 344:348 the 24-residue LRR motif 325:348 the 24-residue LRR motif 325:348 Asparagines conserved in the 24-residue LRR motif contribute extensive hydrogen-bonding networks for solenoid stabilization. 15961631 5 8 theme binding 788:794 arg1 site 796:799 an appropriate binding site 773:799 an appropriate binding site for double-stranded RNA 773:823 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 5 9 from interface 659:667 arg1 crystal 676:682 the crystal 672:682 the crystal 672:682 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 3 10 theme hydrogen-bonding 371:386 arg1 networks 388:395 extensive hydrogen-bonding networks 361:395 extensive hydrogen-bonding networks for solenoid stabilization 361:422 Asparagines conserved in the 24-residue LRR motif contribute extensive hydrogen-bonding networks for solenoid stabilization. 15961631 5 11 theme appropriate 776:786 arg1 site 796:799 an appropriate binding site 773:799 an appropriate binding site for double-stranded RNA 773:823 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 5 12 theme conserved 581:589 arg1 residues 599:606 Highly conserved surface residues 574:606 Highly conserved surface residues 574:606 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 1 13 theme Toll-like 67:75 arg1 receptors 77:85 Toll-like receptors 67:85 Toll-like receptors (TLRs) 67:92 Toll-like receptors (TLRs) play key roles in activating immune responses during infection. 15961631 1 13 theme Toll-like 67:75 arg1 TLRs 88:91 TLRs 88:91 TLRs 88:91 Toll-like receptors (TLRs) play key roles in activating immune responses during infection. 15961631 2 14 from angstroms 201:209 arg1 structure 184:192 The human TLR3 ectodomain structure 158:192 The human TLR3 ectodomain structure at 2.1 angstroms 158:209 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 0 15 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain 0:64 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain. 15961631 5 16 theme homodimer 649:657 arg1 interface 659:667 a homodimer interface 647:667 a homodimer interface in the crystal 647:682 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 5 17 theme surface 591:597 arg1 residues 599:606 Highly conserved surface residues 574:606 Highly conserved surface residues 574:606 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 0 18 theme human 21:25 arg1 ectodomain 55:64 human toll-like receptor 3 (TLR3) ectodomain 21:64 human toll-like receptor 3 (TLR3) ectodomain 21:64 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain. 15961631 3 19 theme extensive 361:369 arg1 networks 388:395 extensive hydrogen-bonding networks 361:395 extensive hydrogen-bonding networks for solenoid stabilization 361:422 Asparagines conserved in the 24-residue LRR motif contribute extensive hydrogen-bonding networks for solenoid stabilization. 15961631 0 20 theme receptor 37:44 arg1 ectodomain 55:64 human toll-like receptor 3 (TLR3) ectodomain 21:64 human toll-like receptor 3 (TLR3) ectodomain 21:64 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain. 15961631 5 21 theme charged 719:725 arg1 residues 727:734 positively charged residues 708:734 positively charged residues 708:734 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 0 22 theme toll-like 27:35 arg1 receptor 37:44 toll-like receptor 3 27:46 human toll-like receptor 3 (TLR3) ectodomain 21:64 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain. 15961631 0 22 theme toll-like 27:35 arg1 TLR3 49:52 TLR3 49:52 TLR3 49:52 Crystal structure of human toll-like receptor 3 (TLR3) ectodomain. 15961631 5 23 theme residues 727:734 arg1 patches 697:703 two patches 693:703 two patches of positively charged residues 693:734 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 5 23 theme residues 727:734 arg1 insertion 749:757 a second insertion 740:757 a second insertion 740:757 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 4 24 from role 530:533 arg1 oligomerization 557:571 oligomerization 557:571 oligomerization 557:571 TLR3 is largely masked by carbohydrate, but one face is glycosylation-free, which suggests its potential role in ligand binding and oligomerization. 15961631 4 24 from role 530:533 arg1 binding 545:551 ligand binding 538:551 ligand binding 538:551 TLR3 is largely masked by carbohydrate, but one face is glycosylation-free, which suggests its potential role in ligand binding and oligomerization. 15961631 4 25 theme potential 520:528 arg1 role 530:533 its potential role 516:533 its potential role in ligand binding and oligomerization 516:571 TLR3 is largely masked by carbohydrate, but one face is glycosylation-free, which suggests its potential role in ligand binding and oligomerization. 15961631 5 26 theme double-stranded 805:819 arg1 RNA 821:823 double-stranded RNA 805:823 double-stranded RNA 805:823 Highly conserved surface residues and a TLR3-specific LRR insertion form a homodimer interface in the crystal, whereas two patches of positively charged residues and a second insertion would provide an appropriate binding site for double-stranded RNA. 15961631 3 27 theme solenoid 401:408 arg1 stabilization 410:422 solenoid stabilization 401:422 solenoid stabilization 401:422 Asparagines conserved in the 24-residue LRR motif contribute extensive hydrogen-bonding networks for solenoid stabilization. 15961631 4 28 theme ligand 538:543 arg1 binding 545:551 ligand binding 538:551 ligand binding 538:551 TLR3 is largely masked by carbohydrate, but one face is glycosylation-free, which suggests its potential role in ligand binding and oligomerization. 15961631 2 29 theme ectodomain 173:182 arg1 structure 184:192 The human TLR3 ectodomain structure 158:192 The human TLR3 ectodomain structure at 2.1 angstroms 158:209 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 1 30 theme key 99:101 arg1 roles 103:107 key roles 99:107 key roles 99:107 Toll-like receptors (TLRs) play key roles in activating immune responses during infection. 15961631 2 31 theme TLR3 168:171 arg1 structure 184:192 The human TLR3 ectodomain structure 158:192 The human TLR3 ectodomain structure at 2.1 angstroms 158:209 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 2 32 theme horseshoe-shaped 227:242 arg1 solenoid 244:251 a large horseshoe-shaped solenoid 219:251 a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs) 219:297 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 2 33 theme human 162:166 arg1 structure 184:192 The human TLR3 ectodomain structure 158:192 The human TLR3 ectodomain structure at 2.1 angstroms 158:209 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 15961631 2 34 theme large 221:225 arg1 solenoid 244:251 a large horseshoe-shaped solenoid 219:251 a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs) 219:297 The human TLR3 ectodomain structure at 2.1 angstroms reveals a large horseshoe-shaped solenoid assembled from 23 leucine-rich repeats (LRRs). 16698036 4 0 theme Sachs 765:769 arg1 disease 771:777 adult onset Tay Sachs disease 749:777 adult onset Tay Sachs disease 749:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 1 1 theme Hex 170:172 arg1 A 167:167 Lysosomal beta-hexosaminidase A 137:167 Lysosomal beta-hexosaminidase A (Hex A) 137:175 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 1 1 theme Hex 170:172 arg1 A 174:174 Hex A 170:174 Hex A 170:174 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 8 2 from betaAsp452 1437:1446 arg1 place 1472:1476 their place 1466:1476 their place 1466:1476 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 0 3 theme mutations 87:95 arg1 loss 101:104 loss 101:104 loss of GM2 ganglioside hydrolysis 101:134 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 0 3 theme mutations 87:95 arg1 interpretation 59:72 interpretation 59:72 interpretation of Tay-Sachs mutations 59:95 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 5 4 theme functional 983:992 arg1 site 1001:1004 a functional active site 981:1004 a functional active site 981:1004 The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. 16698036 4 5 theme Tay 761:763 arg1 disease 771:777 adult onset Tay Sachs disease 749:777 adult onset Tay Sachs disease 749:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 7 6 theme N-acetyl-neuraminic 1346:1364 arg1 residue 1371:1377 the N-acetyl-neuraminic acid residue 1342:1377 the N-acetyl-neuraminic acid residue of GM2 1342:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 6 theme N-acetyl-neuraminic 1346:1364 arg1 GM2 1382:1384 GM2 1382:1384 GM2 1382:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 1 7 theme peripheral 249:258 arg1 system 268:273 the central and peripheral nervous system 233:273 the central and peripheral nervous system 233:273 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 3 8 with complex 536:542 arg1 NAG-thiazoline 549:562 NAG-thiazoline 549:562 NAG-thiazoline, (NGT) to 3.25 A resolution 549:590 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 3 8 with complex 536:542 arg1 NGT 566:568 NGT 566:568 NGT 566:568 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 4 9 dep prevents 695:702 arg1 increases 797:805 increases 797:805 increases the residual activity of Hex A to a level above the critical threshold for disease 797:888 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 2 10 theme GM2 292:294 arg1 Accumulation 276:287 Accumulation 276:287 Accumulation of GM2 276:294 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 3 11 from structure 514:522 arg1 complex 536:542 complex 536:542 complex with NAG-thiazoline, (NGT) to 3.25 A resolution 536:590 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 1 12 theme nervous 260:266 arg1 system 268:273 the central and peripheral nervous system 233:273 the central and peripheral nervous system 233:273 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 3 13 theme A 531:531 arg1 structure 514:522 the structure 510:522 the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution 510:590 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 3 13 theme A 531:531 arg1 structure 467:475 the X-ray crystallographic structure 440:475 the X-ray crystallographic structure of Hex A to 2.8 A resolution 440:504 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 3 14 theme A 484:484 arg1 structure 514:522 the structure 510:522 the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution 510:590 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 3 14 theme A 484:484 arg1 structure 467:475 the X-ray crystallographic structure 440:475 the X-ray crystallographic structure of Hex A to 2.8 A resolution 440:504 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 7 15 theme activator 1256:1264 arg1 protein 1266:1272 the GM2 activator protein 1248:1272 the GM2 activator protein 1248:1272 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 0 16 theme ganglioside 113:123 arg1 hydrolysis 125:134 GM2 ganglioside hydrolysis 109:134 GM2 ganglioside hydrolysis 109:134 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 4 17 theme chemical 654:661 arg1 chaperone 663:671 a chemical chaperone 652:671 a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease 652:888 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 4 17 theme chemical 654:661 arg1 NGT 593:595 NGT 593:595 NGT 593:595 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 6 18 theme active 1030:1035 arg1 site 1037:1040 Only the alpha-subunit active site 1007:1040 Only the alpha-subunit active site 1007:1040 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 4 19 theme mechanism-based 600:614 arg1 inhibitor 616:624 a mechanism-based inhibitor 598:624 a mechanism-based inhibitor 598:624 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 4 19 theme mechanism-based 600:614 arg1 NGT 593:595 NGT 593:595 NGT 593:595 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 0 20 theme GM2 109:111 arg1 hydrolysis 125:134 GM2 ganglioside hydrolysis 109:134 GM2 ganglioside hydrolysis 109:134 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 10 21 theme protein 1750:1756 arg1 function 1758:1765 protein function 1750:1765 protein function 1750:1765 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 4 22 theme critical 859:866 arg1 threshold 868:876 the critical threshold 855:876 the critical threshold for disease 855:888 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 6 23 theme alpha-subunit 1016:1028 arg1 site 1037:1040 Only the alpha-subunit active site 1007:1040 Only the alpha-subunit active site 1007:1040 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 10 24 from effect 1740:1745 arg1 A 1730:1730 a mutant Hex A 1717:1730 a mutant Hex A 1717:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 24 from effect 1740:1745 arg1 site 1709:1712 the active site 1698:1712 the active site of a mutant Hex A 1698:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 24 from effect 1740:1745 arg1 function 1758:1765 protein function 1750:1765 protein function 1750:1765 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 4 25 theme A 836:836 arg1 activity 820:827 the residual activity 807:827 the residual activity of Hex A to a level above the critical threshold for disease 807:888 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 10 26 theme mutant 1719:1724 arg1 A 1730:1730 a mutant Hex A 1717:1730 a mutant Hex A 1717:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 8 27 theme key 1416:1418 arg1 residues 1420:1427 these key residues 1410:1427 these key residues 1410:1427 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 4 28 theme onset 755:759 arg1 disease 771:777 adult onset Tay Sachs disease 749:777 adult onset Tay Sachs disease 749:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 3 29 from structure 467:475 arg1 complex 536:542 complex 536:542 complex with NAG-thiazoline, (NGT) to 3.25 A resolution 536:590 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 2 30 theme debilitating 314:325 arg1 neurodegeneration 327:343 severely debilitating neurodegeneration 305:343 severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant 305:420 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 2 31 theme Sandoff 386:392 arg1 disease 394:400 Sandoff disease 386:400 Sandoff disease (SD) 386:405 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 2 31 theme Sandoff 386:392 arg1 SD 403:404 SD 403:404 SD 403:404 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 0 32 theme Crystallographic 0:15 arg1 structure 17:25 Crystallographic structure 0:25 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 0:135 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 9 33 from Mutations 1551:1559 arg1 those 1608:1612 those 1608:1612 those 1608:1612 Mutations in the alpha-subunit, associated with TSD, and those in the beta-subunit, associated with SD are discussed. 16698036 9 33 from Mutations 1551:1559 arg1 alpha-subunit 1568:1580 the alpha-subunit 1564:1580 the alpha-subunit 1564:1580 Mutations in the alpha-subunit, associated with TSD, and those in the beta-subunit, associated with SD are discussed. 16698036 4 34 theme adult 749:753 arg1 disease 771:777 adult onset Tay Sachs disease 749:777 adult onset Tay Sachs disease 749:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 10 35 theme Hex 1726:1728 arg1 A 1730:1730 a mutant Hex A 1717:1730 a mutant Hex A 1717:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 0 36 theme human 30:34 arg1 A 56:56 human beta-hexosaminidase A 30:56 human beta-hexosaminidase A 30:56 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 6 37 theme GM2 1056:1058 arg1 gangliosides 1060:1071 GM2 gangliosides 1056:1071 GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424 1056:1203 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 6 38 theme alphaArg424 1193:1203 arg1 presence 1165:1172 the presence 1161:1172 the presence of alphaAsn423 and alphaArg424 1161:1203 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 5 39 theme crystal 895:901 arg1 structure 903:911 The crystal structure 891:911 The crystal structure of Hex A 891:920 The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. 16698036 6 40 theme loop 1091:1094 arg1 structure 1096:1104 a flexible loop structure 1080:1104 a flexible loop structure that is removed post-translationally from beta 1080:1151 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 2 41 theme AB 411:412 arg1 variant 414:420 AB variant 411:420 AB variant 411:420 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 0 42 theme hydrolysis 125:134 arg1 loss 101:104 loss 101:104 loss of GM2 ganglioside hydrolysis 101:134 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 0 42 theme hydrolysis 125:134 arg1 interpretation 59:72 interpretation 59:72 interpretation of Tay-Sachs mutations 59:95 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 3 43 theme Hex 527:529 arg1 A 531:531 Hex A 527:531 Hex A 527:531 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 3 44 theme X-ray 444:448 arg1 structure 467:475 the X-ray crystallographic structure 440:475 the X-ray crystallographic structure of Hex A to 2.8 A resolution 440:504 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 0 45 theme A 56:56 arg1 structure 17:25 Crystallographic structure 0:25 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 0:135 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 6 46 attach removed 1114:1120 arg2 structure 1096:1104 a flexible loop structure 1080:1104 a flexible loop structure that is removed post-translationally from beta 1080:1151 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 6 46 attach removed 1114:1120 arg1 beta 1148:1151 beta 1148:1151 beta 1148:1151 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 4 47 theme residual 811:818 arg1 activity 820:827 the residual activity 807:827 the residual activity of Hex A to a level above the critical threshold for disease 807:888 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 10 48 theme active 1702:1707 arg1 A 1730:1730 a mutant Hex A 1717:1730 a mutant Hex A 1717:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 48 theme active 1702:1707 arg1 site 1709:1712 the active site 1698:1712 the active site of a mutant Hex A 1698:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 4 49 theme Hex 720:722 arg1 mutant 726:731 a Hex A mutant 718:731 a Hex A mutant associated with adult onset Tay Sachs disease 718:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 0 50 theme beta-hexosaminidase 36:54 arg1 A 56:56 human beta-hexosaminidase A 30:56 human beta-hexosaminidase A 30:56 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 7 51 theme carboxylate 1321:1331 arg1 residue 1371:1377 the N-acetyl-neuraminic acid residue 1342:1377 the N-acetyl-neuraminic acid residue of GM2 1342:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 51 theme carboxylate 1321:1331 arg1 group 1333:1337 the carboxylate group 1317:1337 the carboxylate group of the N-acetyl-neuraminic acid residue of GM2 1317:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 51 theme carboxylate 1321:1331 arg1 GM2 1382:1384 GM2 1382:1384 GM2 1382:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 8 52 from betaLeu453 1452:1461 arg1 place 1472:1476 their place 1466:1476 their place 1466:1476 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 5 53 theme alphabeta 933:941 arg1 heterodimer 943:953 an alphabeta heterodimer 930:953 an alphabeta heterodimer 930:953 The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. 16698036 6 54 theme flexible 1082:1089 arg1 structure 1096:1104 a flexible loop structure 1080:1104 a flexible loop structure that is removed post-translationally from beta 1080:1151 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 1 55 theme GM2 213:215 arg1 gangliosides 217:228 GM2 gangliosides 213:228 GM2 gangliosides 213:228 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 7 56 theme acid 1366:1369 arg1 residue 1371:1377 the N-acetyl-neuraminic acid residue 1342:1377 the N-acetyl-neuraminic acid residue of GM2 1342:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 56 theme acid 1366:1369 arg1 GM2 1382:1384 GM2 1382:1384 GM2 1382:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 6 57 theme alphaAsn423 1177:1187 arg1 presence 1165:1172 the presence 1161:1172 the presence of alphaAsn423 and alphaArg424 1161:1203 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 4 58 theme mutant 726:731 arg1 misfolding 704:713 misfolding 704:713 misfolding of a Hex A mutant associated with adult onset Tay Sachs disease 704:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 4 58 theme mutant 726:731 arg1 result 789:794 a result 787:794 a result 787:794 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 10 59 from effect 1673:1678 arg1 A 1730:1730 a mutant Hex A 1717:1730 a mutant Hex A 1717:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 59 from effect 1673:1678 arg1 site 1709:1712 the active site 1698:1712 the active site of a mutant Hex A 1698:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 59 from effect 1673:1678 arg1 function 1758:1765 protein function 1750:1765 protein function 1750:1765 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 2 60 theme Tay-Sachs 361:369 arg1 TSD 380:382 TSD 380:382 TSD 380:382 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 2 60 theme Tay-Sachs 361:369 arg1 disease 371:377 Tay-Sachs disease 361:377 Tay-Sachs disease (TSD) 361:383 Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. 16698036 5 61 theme active 994:999 arg1 site 1001:1004 a functional active site 981:1004 a functional active site 981:1004 The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. 16698036 10 62 theme binding 1687:1693 arg1 effect 1740:1745 its effect 1736:1745 its effect on protein function 1736:1765 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 62 theme binding 1687:1693 arg1 effect 1673:1678 The effect 1669:1678 The effect of NGT binding in the active site of a mutant Hex A 1669:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 7 63 theme residue 1371:1377 arg1 residue 1371:1377 the N-acetyl-neuraminic acid residue 1342:1377 the N-acetyl-neuraminic acid residue of GM2 1342:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 63 theme residue 1371:1377 arg1 group 1333:1337 the carboxylate group 1317:1337 the carboxylate group of the N-acetyl-neuraminic acid residue of GM2 1317:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 63 theme residue 1371:1377 arg1 GM2 1382:1384 GM2 1382:1384 GM2 1382:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 4 64 theme A 724:724 arg1 mutant 726:731 a Hex A mutant 718:731 a Hex A mutant associated with adult onset Tay Sachs disease 718:777 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 10 65 theme A 1730:1730 arg1 A 1730:1730 a mutant Hex A 1717:1730 a mutant Hex A 1717:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 10 65 theme A 1730:1730 arg1 site 1709:1712 the active site 1698:1712 the active site of a mutant Hex A 1698:1730 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 3 66 theme Hex 480:482 arg1 A 484:484 Hex A 480:484 Hex A 480:484 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 7 67 theme loop 1210:1213 arg1 structure 1215:1223 The loop structure 1206:1223 The loop structure 1206:1223 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 6 68 theme due 1073:1075 arg1 gangliosides 1060:1071 GM2 gangliosides 1056:1071 GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424 1056:1203 Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. 16698036 1 69 from degradation 198:208 arg1 system 268:273 the central and peripheral nervous system 233:273 the central and peripheral nervous system 233:273 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 1 70 theme Lysosomal 137:145 arg1 A 167:167 Lysosomal beta-hexosaminidase A 137:167 Lysosomal beta-hexosaminidase A (Hex A) 137:175 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 1 70 theme Lysosomal 137:145 arg1 A 174:174 Hex A 170:174 Hex A 170:174 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 1 71 theme gangliosides 217:228 arg1 degradation 198:208 the degradation 194:208 the degradation of GM2 gangliosides in the central and peripheral nervous system 194:273 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 8 72 contain has 1433:1435 arg2 betaLeu453 1452:1461 betaLeu453 1452:1461 betaLeu453 1452:1461 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 8 72 contain has 1433:1435 arg2 betaAsp452 1437:1446 betaAsp452 1437:1446 betaAsp452 1437:1446 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 8 72 contain has 1433:1435 arg1 beta-subunit 1391:1402 The beta-subunit 1387:1402 The beta-subunit 1387:1402 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 5 73 theme Hex 916:918 arg1 A 920:920 Hex A 916:920 Hex A 916:920 The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. 16698036 10 74 theme NGT 1683:1685 arg1 binding 1687:1693 NGT binding 1683:1693 NGT binding 1683:1693 The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. 16698036 1 75 theme beta-hexosaminidase 147:165 arg1 A 167:167 Lysosomal beta-hexosaminidase A 137:167 Lysosomal beta-hexosaminidase A (Hex A) 137:175 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 1 75 theme beta-hexosaminidase 147:165 arg1 A 174:174 Hex A 170:174 Hex A 170:174 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 16698036 3 76 theme crystallographic 450:465 arg1 structure 467:475 the X-ray crystallographic structure 440:475 the X-ray crystallographic structure of Hex A to 2.8 A resolution 440:504 Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. 16698036 0 77 theme Tay-Sachs 77:85 arg1 mutations 87:95 Tay-Sachs mutations 77:95 Tay-Sachs mutations 77:95 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 5 78 theme A 920:920 arg1 structure 903:911 The crystal structure 891:911 The crystal structure of Hex A 891:920 The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. 16698036 0 79 dep structure 17:25 arg1 loss 101:104 loss 101:104 loss of GM2 ganglioside hydrolysis 101:134 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 0 79 dep structure 17:25 arg1 interpretation 59:72 interpretation 59:72 interpretation of Tay-Sachs mutations 59:95 Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay-Sachs mutations and loss of GM2 ganglioside hydrolysis. 16698036 8 80 theme neutral 1519:1525 arg1 substrates 1527:1536 only neutral substrates 1514:1536 only neutral substrates 1514:1536 The beta-subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. 16698036 7 81 theme GM2 1382:1384 arg1 residue 1371:1377 the N-acetyl-neuraminic acid residue 1342:1377 the N-acetyl-neuraminic acid residue of GM2 1342:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 7 81 theme GM2 1382:1384 arg1 GM2 1382:1384 GM2 1382:1384 GM2 1382:1384 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 4 82 theme Hex 832:834 arg1 A 836:836 Hex A 832:836 Hex A 832:836 NGT, a mechanism-based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. 16698036 7 83 theme GM2 1252:1254 arg1 protein 1266:1272 the GM2 activator protein 1248:1272 the GM2 activator protein 1248:1272 The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. 16698036 1 84 theme central 237:243 arg1 system 268:273 the central and peripheral nervous system 233:273 the central and peripheral nervous system 233:273 Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. 19654028 1 0 with receptor 145:152 arg1 function 206:213 an apparent regulatory function 183:213 an apparent regulatory function in hematopoietic and neuronal cells 183:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 2 1 theme mobility 482:489 arg1 shift 491:495 its approximately 40 kDa mobility shift 457:495 its approximately 40 kDa mobility shift in SDS polyacrylamide gels 457:522 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 6 2 theme DR6 1293:1295 arg1 targeting 1297:1305 DR6 targeting 1293:1305 DR6 targeting into Brij-98 insoluble lipid rafts 1293:1340 However, palmitoylation of Cys368 is apparently not required for DR6 targeting into Brij-98 insoluble lipid rafts. 19654028 3 3 theme " 665:665 arg1 domain 667:672 the "stalk" domain 655:672 the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) 655:719 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 4 4 theme DR6 952:954 arg1 localization 936:947 the plasma membrane localization 916:947 the plasma membrane localization of DR6 916:954 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 2 5 dep kDa 478:480 arg1 40 475:476 40 475:476 40 475:476 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 5 6 theme DR4 1201:1203 arg1 ICPs 1205:1208 Fas/CD95 or DR4 ICPs 1189:1208 Fas/CD95 or DR4 ICPs 1189:1208 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 7 7 theme membrane 1457:1464 arg1 microdomains 1466:1477 these membrane microdomains 1451:1477 these membrane microdomains 1451:1477 In contrast, we show that N-glycosylation of the extracellular part might participate in directing DR6 into these membrane microdomains. 19654028 3 8 theme major 726:730 arg1 cluster 644:650 the Ser/Thr/Pro cluster 628:650 the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) 628:719 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 3 8 theme major 726:730 arg1 site 732:735 a major site 724:735 a major site for the likely mucine-type of O-glycosylation 724:781 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 5 9 from cysteines 1176:1184 arg1 ICPs 1205:1208 Fas/CD95 or DR4 ICPs 1189:1208 Fas/CD95 or DR4 ICPs 1189:1208 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 5 10 theme intracellular 1133:1145 arg1 part 1147:1150 the intracellular part 1129:1150 the intracellular part of DR6 1129:1157 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 4 11 theme transmembrane 842:854 arg1 domain 856:861 the transmembrane domain 838:861 the transmembrane domain 838:861 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 2 12 theme extracellular 411:423 arg1 part 425:428 its extracellular part 407:428 its extracellular part 407:428 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 3 13 from cluster 644:650 arg1 domain 667:672 the "stalk" domain 655:672 the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) 655:719 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 2 14 theme acids 398:402 arg1 N- 365:366 N- 365:366 N- 365:366 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 2 15 theme polyacrylamide 504:517 arg1 gels 519:522 SDS polyacrylamide gels 500:522 SDS polyacrylamide gels 500:522 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 4 16 theme amino 882:886 arg1 acids 888:892 130 amino acids 878:892 130 amino acids 878:892 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 7 17 gly N-glycosylation 1369:1383 arg1 part 1406:1409 the extracellular part 1388:1409 the extracellular part 1388:1409 In contrast, we show that N-glycosylation of the extracellular part might participate in directing DR6 into these membrane microdomains. 19654028 5 18 with mutagenesis 1066:1076 arg1 palmitate 1038:1046 radiolabeled palmitate 1025:1046 radiolabeled palmitate 1025:1046 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 2 19 theme amino 392:396 arg1 acids 398:402 amino acids 392:402 amino acids in its extracellular part 392:428 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 4 20 theme intracellular 973:985 arg1 retention 987:995 its intracellular retention 969:995 its intracellular retention 969:995 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 2 21 theme SDS 500:502 arg1 gels 519:522 SDS polyacrylamide gels 500:522 SDS polyacrylamide gels 500:522 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 1 22 theme Death 84:88 arg1 receptor 90:97 Death receptor 6 84:99 Death receptor 6 (DR6/TNFRSF21) 84:114 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 1 22 theme Death 84:88 arg1 DR6/TNFRSF21 102:113 DR6/TNFRSF21 102:113 DR6/TNFRSF21 102:113 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 1 22 theme Death 84:88 arg1 receptor 145:152 a death domain-containing receptor 119:152 a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells 119:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 5 23 theme Fas/CD95 1189:1196 arg1 ICPs 1205:1208 Fas/CD95 or DR4 ICPs 1189:1208 Fas/CD95 or DR4 ICPs 1189:1208 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 1 24 theme superfamily 166:176 arg1 receptor 90:97 Death receptor 6 84:99 Death receptor 6 (DR6/TNFRSF21) 84:114 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 1 24 theme superfamily 166:176 arg1 receptor 145:152 a death domain-containing receptor 119:152 a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells 119:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 0 25 theme Functional 0:9 arg1 analysis 11:18 Functional analysis 0:18 Functional analysis of the posttranslational modifications of the death receptor 6 0:81 Functional analysis of the posttranslational modifications of the death receptor 6. 19654028 3 26 theme likely 745:750 arg1 mucine-type 752:762 the likely mucine-type 741:762 the likely mucine-type of O-glycosylation 741:781 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 4 27 theme linker 807:812 arg1 region 814:819 the entire linker region 796:819 the entire linker region 796:819 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 4 28 theme plasma 920:925 arg1 localization 936:947 the plasma membrane localization 916:947 the plasma membrane localization of DR6 916:954 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 2 29 from N- 365:366 arg1 part 425:428 its extracellular part 407:428 its extracellular part 407:428 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 7 30 theme part 1406:1409 arg1 N-glycosylation 1369:1383 N-glycosylation 1369:1383 N-glycosylation of the extracellular part 1369:1409 In contrast, we show that N-glycosylation of the extracellular part might participate in directing DR6 into these membrane microdomains. 19654028 2 31 from shift 491:495 arg1 gels 519:522 SDS polyacrylamide gels 500:522 SDS polyacrylamide gels 500:522 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 3 32 theme extracellular 574:586 arg1 asparagines 588:598 all six extracellular asparagines 566:598 all six extracellular asparagines 566:598 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 1 33 theme apparent 186:193 arg1 function 206:213 an apparent regulatory function 183:213 an apparent regulatory function in hematopoietic and neuronal cells 183:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 0 34 theme modifications 45:57 arg1 analysis 11:18 Functional analysis 0:18 Functional analysis of the posttranslational modifications of the death receptor 6 0:81 Functional analysis of the posttranslational modifications of the death receptor 6. 19654028 5 35 theme membrane-proximal 1101:1117 arg1 S-palmitoylated 1211:1225 S-palmitoylated 1211:1225 S-palmitoylated 1211:1225 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 5 35 theme membrane-proximal 1101:1117 arg1 Cys368 1119:1124 the membrane-proximal Cys368 1097:1124 the membrane-proximal Cys368 in the intracellular part of DR6 1097:1157 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 5 36 theme radiolabeled 1025:1036 arg1 palmitate 1038:1046 radiolabeled palmitate 1025:1046 radiolabeled palmitate 1025:1046 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 1 37 theme regulatory 195:204 arg1 function 206:213 an apparent regulatory function 183:213 an apparent regulatory function in hematopoietic and neuronal cells 183:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 0 38 theme posttranslational 27:43 arg1 modifications 45:57 the posttranslational modifications 23:57 the posttranslational modifications of the death receptor 6 23:81 Functional analysis of the posttranslational modifications of the death receptor 6. 19654028 4 39 theme membrane 927:934 arg1 localization 936:947 the plasma membrane localization 916:947 the plasma membrane localization of DR6 916:954 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 4 40 theme region 814:819 arg1 Deletion 784:791 Deletion 784:791 Deletion 784:791 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 7 41 theme extracellular 1392:1404 arg1 part 1406:1409 the extracellular part 1388:1409 the extracellular part 1388:1409 In contrast, we show that N-glycosylation of the extracellular part might participate in directing DR6 into these membrane microdomains. 19654028 3 42 gly N-glycosylated 604:617 arg1 asparagines 588:598 all six extracellular asparagines 566:598 all six extracellular asparagines 566:598 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 5 43 from Cys368 1119:1124 arg1 part 1147:1150 the intracellular part 1129:1150 the intracellular part of DR6 1129:1157 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 2 44 dep shift 491:495 arg1 kDa 478:480 kDa 478:480 kDa 478:480 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 3 45 theme stalk 660:664 arg1 domain 667:672 the "stalk" domain 655:672 the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) 655:719 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 1 46 from function 206:213 arg1 cells 245:249 hematopoietic and neuronal cells 218:249 cells 245:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 5 47 theme side-directed 1052:1064 arg1 mutagenesis 1066:1076 side-directed mutagenesis 1052:1076 side-directed mutagenesis 1052:1076 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 2 48 from acids 398:402 arg1 part 425:428 its extracellular part 407:428 its extracellular part 407:428 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 2 49 theme transmembrane 334:346 arg1 protein 348:354 an extensively posttranslationally modified transmembrane protein 290:354 an extensively posttranslationally modified transmembrane protein 290:354 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 2 49 theme transmembrane 334:346 arg1 DR6 283:285 DR6 283:285 DR6 283:285 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 3 50 theme O-glycosylation 767:781 arg1 mucine-type 752:762 the likely mucine-type 741:762 the likely mucine-type of O-glycosylation 741:781 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 6 51 theme lipid 1330:1334 arg1 rafts 1336:1340 Brij-98 insoluble lipid rafts 1312:1340 Brij-98 insoluble lipid rafts 1312:1340 However, palmitoylation of Cys368 is apparently not required for DR6 targeting into Brij-98 insoluble lipid rafts. 19654028 3 52 theme cysteine-rich 692:704 arg1 CRDs 715:718 CRDs 715:718 CRDs 715:718 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 3 52 theme cysteine-rich 692:704 arg1 domains 706:712 the cysteine-rich domains 688:712 the cysteine-rich domains (CRDs) 688:719 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 6 53 theme Cys368 1255:1260 arg1 palmitoylation 1237:1250 palmitoylation 1237:1250 palmitoylation of Cys368 1237:1260 However, palmitoylation of Cys368 is apparently not required for DR6 targeting into Brij-98 insoluble lipid rafts. 19654028 2 54 theme modified 325:332 arg1 protein 348:354 an extensively posttranslationally modified transmembrane protein 290:354 an extensively posttranslationally modified transmembrane protein 290:354 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 2 54 theme modified 325:332 arg1 DR6 283:285 DR6 283:285 DR6 283:285 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 6 55 theme insoluble 1320:1328 arg1 rafts 1336:1340 Brij-98 insoluble lipid rafts 1312:1340 Brij-98 insoluble lipid rafts 1312:1340 However, palmitoylation of Cys368 is apparently not required for DR6 targeting into Brij-98 insoluble lipid rafts. 19654028 5 56 theme Biosynthetic 998:1009 arg1 labeling 1011:1018 Biosynthetic labeling 998:1018 Biosynthetic labeling with radiolabeled palmitate 998:1046 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 3 57 theme Site-directed 525:537 arg1 mutagenesis 539:549 Site-directed mutagenesis 525:549 Site-directed mutagenesis 525:549 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 1 58 theme hematopoietic 218:230 arg1 cells 245:249 hematopoietic and neuronal cells 218:249 cells 245:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 0 59 theme receptor 72:79 arg1 modifications 45:57 the posttranslational modifications 23:57 the posttranslational modifications of the death receptor 6 23:81 Functional analysis of the posttranslational modifications of the death receptor 6. 19654028 3 60 theme Ser/Thr/Pro 632:642 arg1 cluster 644:650 the Ser/Thr/Pro cluster 628:650 the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) 628:719 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 3 60 theme Ser/Thr/Pro 632:642 arg1 site 732:735 a major site 724:735 a major site for the likely mucine-type of O-glycosylation 724:781 Site-directed mutagenesis confirmed that all six extracellular asparagines are N-glycosylated and that the Ser/Thr/Pro cluster in the "stalk" domain juxtaposed to the cysteine-rich domains (CRDs) is a major site for the likely mucine-type of O-glycosylation. 19654028 1 61 theme death 121:125 arg1 receptor 90:97 Death receptor 6 84:99 Death receptor 6 (DR6/TNFRSF21) 84:114 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 1 61 theme death 121:125 arg1 receptor 145:152 a death domain-containing receptor 119:152 a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells 119:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 1 62 theme TNFR 161:164 arg1 superfamily 166:176 the TNFR superfamily 157:176 the TNFR superfamily 157:176 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 0 63 theme death 66:70 arg1 receptor 72:79 the death receptor 6 62:81 the death receptor 6 62:81 Functional analysis of the posttranslational modifications of the death receptor 6. 19654028 5 64 theme DR6 1155:1157 arg1 part 1147:1150 the intracellular part 1129:1150 the intracellular part of DR6 1129:1157 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 4 65 theme entire 800:805 arg1 region 814:819 the entire linker region 796:819 the entire linker region 796:819 Deletion of the entire linker region between CRDs and the transmembrane domain, spanning over 130 amino acids, severely compromises the plasma membrane localization of DR6 and leads to its intracellular retention. 19654028 2 66 from part 425:428 arg1 N- 365:366 N- 365:366 N- 365:366 In this study we document that DR6 is an extensively posttranslationally modified transmembrane protein and that N- and O-glycosylations of amino acids in its extracellular part are mainly responsible for its approximately 40 kDa mobility shift in SDS polyacrylamide gels. 19654028 1 67 theme domain-containing 127:143 arg1 receptor 90:97 Death receptor 6 84:99 Death receptor 6 (DR6/TNFRSF21) 84:114 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 1 67 theme domain-containing 127:143 arg1 receptor 145:152 a death domain-containing receptor 119:152 a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells 119:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 19654028 5 68 with labeling 1011:1018 arg1 palmitate 1038:1046 radiolabeled palmitate 1025:1046 radiolabeled palmitate 1025:1046 Biosynthetic labeling with radiolabeled palmitate and side-directed mutagenesis also revealed that the membrane-proximal Cys368 in the intracellular part of DR6 is, similarly as cysteines in Fas/CD95 or DR4 ICPs, S-palmitoylated. 19654028 1 69 theme neuronal 236:243 arg1 cells 245:249 hematopoietic and neuronal cells 218:249 cells 245:249 Death receptor 6 (DR6/TNFRSF21) is a death domain-containing receptor of the TNFR superfamily with an apparent regulatory function in hematopoietic and neuronal cells. 16445295 11 0 theme remaining 1510:1518 arg1 cystines 1524:1531 The remaining six cystines 1506:1531 The remaining six cystines 1506:1531 The remaining six cystines form disulfides, including Cys156-Cys169, Cys228-Cys252, and Cys243-Cys257. 16445295 3 1 theme TAFI 462:465 arg1 pI 455:456 pI 455:456 pI 455:456 During this study, we observed that (i) the isoelectric point (pI) of TAFI shifts dramatically from pH 5 toward pH 8 upon activation and (ii) TAFIa is significantly less soluble than TAFI. 16445295 3 1 theme TAFI 462:465 arg1 point 448:452 the isoelectric point 432:452 the isoelectric point (pI) of TAFI 432:465 During this study, we observed that (i) the isoelectric point (pI) of TAFI shifts dramatically from pH 5 toward pH 8 upon activation and (ii) TAFIa is significantly less soluble than TAFI. 16445295 8 2 located detected 1220:1227 arg1 addition 1157:1164 addition 1157:1164 addition 1157:1164 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 8 2 located detected 1220:1227 arg2 GlcNAc 1177:1182 bisecting GlcNAc 1167:1182 bisecting GlcNAc attached to the trimannose core 1167:1214 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 1 3 theme procarboxypeptidase 263:281 arg1 B 283:283 pancreatic procarboxypeptidase B 252:283 pancreatic procarboxypeptidase B in several ways 252:299 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 14 4 theme TAFIa 1992:1996 arg1 instability 1977:1987 the inherent instability 1964:1987 the inherent instability of TAFIa 1964:1996 When these results are taken together, they suggest that the inherent instability of TAFIa is not caused by post-translational modifications. 16445295 6 5 located found 972:976 arg2 Asn219 956:961 Asn219 956:961 Asn219 956:961 Asn219 was also found in an unglycosylated variant. 16445295 6 5 located found 972:976 arg1 variant 999:1005 an unglycosylated variant 981:1005 an unglycosylated variant 981:1005 Asn219 was also found in an unglycosylated variant. 16445295 3 6 dep TAFIa 534:538 arg1 ii 530:531 ii 530:531 ii 530:531 During this study, we observed that (i) the isoelectric point (pI) of TAFI shifts dramatically from pH 5 toward pH 8 upon activation and (ii) TAFIa is significantly less soluble than TAFI. 16445295 15 7 theme attached 2099:2106 arg1 glycans 2108:2114 the attached glycans 2095:2114 the attached glycans 2095:2114 However, after activation, TAFIa loses 80% of the attached glycans, generating a large shift in pI and a propensity to precipitate. 16445295 5 8 theme catalytic 937:945 arg1 domain 947:952 the catalytic domain 933:952 the catalytic domain 933:952 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 8 9 theme other 1251:1255 arg1 origin 1244:1249 an origin 1241:1249 an origin other than the liver 1241:1270 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 13 10 theme additional 1881:1890 arg1 modifications 1892:1904 no additional modifications 1878:1904 no additional modifications 1878:1904 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 2 11 theme enzymatic 306:314 arg1 unstable 337:344 unstable 337:344 unstable 337:344 The enzymatic activity of TAFIa is unstable and decays with a half-life of a few minutes. 16445295 2 11 theme enzymatic 306:314 arg1 activity 316:323 The enzymatic activity 302:323 The enzymatic activity of TAFIa 302:332 The enzymatic activity of TAFIa is unstable and decays with a half-life of a few minutes. 16445295 10 12 contain contain 1474:1480 arg1 Cys69 1421:1425 Cys69 1421:1425 Cys69 1421:1425 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 10 12 contain contain 1474:1480 arg1 two 1416:1418 two 1416:1418 two 1416:1418 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 10 12 contain contain 1474:1480 arg1 Cys383 1431:1436 Cys383 1431:1436 Cys383 1431:1436 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 10 12 contain contain 1474:1480 arg2 groups 1498:1503 free sulfhydryl groups 1482:1503 free sulfhydryl groups 1482:1503 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 13 13 theme acid 1855:1858 arg1 sequence 1860:1867 the TAFI amino acid sequence 1840:1867 the TAFI amino acid sequence 1840:1867 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 5 14 used utilized 827:834 arg2 sites 816:820 all five potential N-glycosylation sites 781:820 all five potential N-glycosylation sites 781:820 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 5 15 from domain 947:952 arg1 located 922:928 located 922:928 located 922:928 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 1 16 theme several 288:294 arg1 ways 296:299 several ways 288:299 several ways 288:299 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 9 17 dep composition 1288:1298 arg1 analyses 1313:1320 analyses 1313:1320 analyses 1313:1320 Monosaccharide composition and LC-MS/MS analyses did not produce evidence for O glycosylation. 16445295 14 18 theme post-translational 2015:2032 arg1 modifications 2034:2046 post-translational modifications 2015:2046 post-translational modifications 2015:2046 When these results are taken together, they suggest that the inherent instability of TAFIa is not caused by post-translational modifications. 16445295 16 19 theme TAFIa 2248:2252 arg1 properties 2234:2243 the properties 2230:2243 the properties of TAFIa 2230:2252 These changes are likely to significantly affect the properties of TAFIa as compared to TAFI. 16445295 2 20 theme minutes 383:389 arg1 half-life 364:372 a half-life 362:372 a half-life of a few minutes 362:389 The enzymatic activity of TAFIa is unstable and decays with a half-life of a few minutes. 16445295 9 21 theme O 1351:1351 arg1 glycosylation 1353:1365 O glycosylation 1351:1365 O glycosylation 1351:1365 Monosaccharide composition and LC-MS/MS analyses did not produce evidence for O glycosylation. 16445295 0 22 theme large 109:113 arg1 shift 115:119 a large shift 107:119 a large shift in the isoelectric point 107:144 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 10 23 theme cysteine 1388:1395 arg1 residues 1397:1404 eight cysteine residues 1382:1404 eight cysteine residues 1382:1404 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 12 24 theme procarboxypeptidase 1650:1668 arg1 B 1670:1670 pancreatic procarboxypeptidase B 1639:1670 pancreatic procarboxypeptidase B 1639:1670 This pattern is homologous to pancreatic procarboxypeptidase B, and it is therefore unlikely that permutations in the cysteine connectivity are responsible for the enzymatic instability. 16445295 13 25 theme LC-MS/MS 1796:1803 arg1 analyses 1805:1812 LC-MS/MS analyses 1796:1812 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence 1796:1867 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 4 26 theme structural 585:594 arg1 bases 596:600 The structural bases 581:600 The structural bases for these observations 581:623 The structural bases for these observations were investigated by characterizing all post-translational modifications, including attached glycans and disulfide connectivity. 16445295 8 27 attach attached 1184:1191 arg1 core 1211:1214 the trimannose core 1196:1214 the trimannose core 1196:1214 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 8 27 attach attached 1184:1191 arg2 GlcNAc 1177:1182 bisecting GlcNAc 1167:1182 bisecting GlcNAc attached to the trimannose core 1167:1214 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 12 28 theme pancreatic 1639:1648 arg1 B 1670:1670 pancreatic procarboxypeptidase B 1639:1670 pancreatic procarboxypeptidase B 1639:1670 This pattern is homologous to pancreatic procarboxypeptidase B, and it is therefore unlikely that permutations in the cysteine connectivity are responsible for the enzymatic instability. 16445295 5 29 dep Asn86 867:871 arg1 located 874:880 located 874:880 located 874:880 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 10 30 theme sulfhydryl 1487:1496 arg1 groups 1498:1503 free sulfhydryl groups 1482:1503 free sulfhydryl groups 1482:1503 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 1 31 theme Thrombin-activable 186:203 arg1 TAFI 229:232 TAFI 229:232 TAFI 229:232 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 1 31 theme Thrombin-activable 186:203 arg1 inhibitor 218:226 Thrombin-activable fibrinolysis inhibitor 186:226 Thrombin-activable fibrinolysis inhibitor (TAFI) 186:233 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 0 32 theme Post-translational 0:17 arg1 evidence 94:101 evidence 94:101 evidence for a large shift in the isoelectric point 94:144 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 32 theme Post-translational 0:17 arg1 solubility 158:167 reduced solubility 150:167 reduced solubility upon activation 150:183 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 32 theme Post-translational 0:17 arg1 modifications 19:31 Post-translational modifications 0:31 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 0:184 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 7 33 theme glycans 1020:1026 arg1 Asn63 1036:1040 Asn63 1036:1040 Asn63 1036:1040 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 33 theme glycans 1020:1026 arg1 Asn86 1043:1047 Asn86 1043:1047 Asn86 1043:1047 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 33 theme glycans 1020:1026 arg1 glycans 1020:1026 the glycans 1016:1026 the glycans 1016:1026 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 33 theme glycans 1020:1026 arg1 Asn51 1029:1033 Asn51 1029:1033 Asn51 1029:1033 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 33 theme glycans 1020:1026 arg1 Asn219 1054:1059 Asn219 1054:1059 Asn219 1054:1059 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 33 theme glycans 1020:1026 arg1 Four 1008:1011 Four 1008:1011 Four 1008:1011 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 5 34 from located 874:880 arg1 peptide 900:906 the activation peptide 885:906 the activation peptide 885:906 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 2 35 theme TAFIa 328:332 arg1 unstable 337:344 unstable 337:344 unstable 337:344 The enzymatic activity of TAFIa is unstable and decays with a half-life of a few minutes. 16445295 2 35 theme TAFIa 328:332 arg1 activity 316:323 The enzymatic activity 302:323 The enzymatic activity of TAFIa 302:332 The enzymatic activity of TAFIa is unstable and decays with a half-life of a few minutes. 16445295 0 36 theme inhibitor 76:84 arg1 evidence 94:101 evidence 94:101 evidence for a large shift in the isoelectric point 94:144 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 36 theme inhibitor 76:84 arg1 solubility 158:167 reduced solubility 150:167 reduced solubility upon activation 150:183 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 36 theme inhibitor 76:84 arg1 modifications 19:31 Post-translational modifications 0:31 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 0:184 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 2 37 theme few 379:381 arg1 minutes 383:389 a few minutes 377:389 a few minutes 377:389 The enzymatic activity of TAFIa is unstable and decays with a half-life of a few minutes. 16445295 0 38 theme human 36:40 arg1 TAFI 87:90 TAFI 87:90 TAFI 87:90 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 38 theme human 36:40 arg1 inhibitor 76:84 human thrombin-activatable fibrinolysis inhibitor 36:84 human thrombin-activatable fibrinolysis inhibitor (TAFI) 36:91 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 13 39 theme sequence 1860:1867 arg1 sequence 1860:1867 the TAFI amino acid sequence 1840:1867 the TAFI amino acid sequence 1840:1867 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 13 39 theme sequence 1860:1867 arg1 % 1835:1835 more than 90% 1823:1835 more than 90% of the TAFI amino acid sequence 1823:1867 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 5 40 theme activation 889:898 arg1 peptide 900:906 the activation peptide 885:906 the activation peptide 885:906 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 3 41 theme isoelectric 436:446 arg1 pI 455:456 pI 455:456 pI 455:456 During this study, we observed that (i) the isoelectric point (pI) of TAFI shifts dramatically from pH 5 toward pH 8 upon activation and (ii) TAFIa is significantly less soluble than TAFI. 16445295 3 41 theme isoelectric 436:446 arg1 point 448:452 the isoelectric point 432:452 the isoelectric point (pI) of TAFI 432:465 During this study, we observed that (i) the isoelectric point (pI) of TAFI shifts dramatically from pH 5 toward pH 8 upon activation and (ii) TAFIa is significantly less soluble than TAFI. 16445295 3 42 dep shifts 467:472 arg1 i 429:429 i 429:429 i 429:429 During this study, we observed that (i) the isoelectric point (pI) of TAFI shifts dramatically from pH 5 toward pH 8 upon activation and (ii) TAFIa is significantly less soluble than TAFI. 16445295 6 43 gly unglycosylated 984:997 arg1 variant 999:1005 an unglycosylated variant 981:1005 an unglycosylated variant 981:1005 Asn219 was also found in an unglycosylated variant. 16445295 4 44 theme attached 709:716 arg1 glycans 718:724 attached glycans 709:724 attached glycans 709:724 The structural bases for these observations were investigated by characterizing all post-translational modifications, including attached glycans and disulfide connectivity. 16445295 8 45 theme bisecting 1167:1175 arg1 GlcNAc 1177:1182 bisecting GlcNAc 1167:1182 bisecting GlcNAc attached to the trimannose core 1167:1214 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 0 46 theme fibrinolysis 63:74 arg1 TAFI 87:90 TAFI 87:90 TAFI 87:90 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 46 theme fibrinolysis 63:74 arg1 inhibitor 76:84 human thrombin-activatable fibrinolysis inhibitor 36:84 human thrombin-activatable fibrinolysis inhibitor (TAFI) 36:91 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 10 47 theme free 1482:1485 arg1 groups 1498:1503 free sulfhydryl groups 1482:1503 free sulfhydryl groups 1482:1503 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 1 48 theme fibrinolysis 205:216 arg1 TAFI 229:232 TAFI 229:232 TAFI 229:232 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 1 48 theme fibrinolysis 205:216 arg1 inhibitor 218:226 Thrombin-activable fibrinolysis inhibitor 186:226 Thrombin-activable fibrinolysis inhibitor (TAFI) 186:233 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 0 49 theme thrombin-activatable 42:61 arg1 TAFI 87:90 TAFI 87:90 TAFI 87:90 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 49 theme thrombin-activatable 42:61 arg1 inhibitor 76:84 human thrombin-activatable fibrinolysis inhibitor 36:84 human thrombin-activatable fibrinolysis inhibitor (TAFI) 36:91 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 1 50 from B 283:283 arg1 ways 296:299 several ways 288:299 several ways 288:299 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 1 50 from B 283:283 arg1 distinct 238:245 distinct 238:245 distinct 238:245 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 16445295 14 51 theme inherent 1968:1975 arg1 instability 1977:1987 the inherent instability 1964:1987 the inherent instability of TAFIa 1964:1996 When these results are taken together, they suggest that the inherent instability of TAFIa is not caused by post-translational modifications. 16445295 5 52 theme potential 790:798 arg1 sites 816:820 all five potential N-glycosylation sites 781:820 all five potential N-glycosylation sites 781:820 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 12 53 theme cysteine 1727:1734 arg1 connectivity 1736:1747 the cysteine connectivity 1723:1747 the cysteine connectivity 1723:1747 This pattern is homologous to pancreatic procarboxypeptidase B, and it is therefore unlikely that permutations in the cysteine connectivity are responsible for the enzymatic instability. 16445295 10 54 contain contains 1373:1380 arg2 residues 1397:1404 eight cysteine residues 1382:1404 eight cysteine residues 1382:1404 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 10 54 contain contains 1373:1380 arg1 TAFI 1368:1371 TAFI 1368:1371 TAFI 1368:1371 TAFI contains eight cysteine residues, of which two, Cys69 and Cys383, are not involved in disulfides and contain free sulfhydryl groups. 16445295 0 55 theme reduced 150:156 arg1 solubility 158:167 reduced solubility 150:167 reduced solubility upon activation 150:183 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 55 theme reduced 150:156 arg1 modifications 19:31 Post-translational modifications 0:31 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 0:184 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 5 56 gly N-glycosylation 800:814 arg2 sites 816:820 all five potential N-glycosylation sites 781:820 all five potential N-glycosylation sites 781:820 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 5 56 gly N-glycosylation 800:814 arg2 five 785:788 five 785:788 five 785:788 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 8 57 theme trimannose 1200:1209 arg1 core 1211:1214 the trimannose core 1196:1214 the trimannose core 1196:1214 In addition, bisecting GlcNAc attached to the trimannose core was detected, suggesting an origin other than the liver. 16445295 0 58 theme isoelectric 128:138 arg1 point 140:144 the isoelectric point 124:144 the isoelectric point 124:144 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 9 59 theme Monosaccharide 1273:1286 arg1 composition 1288:1298 Monosaccharide composition 1273:1298 Monosaccharide composition 1273:1298 Monosaccharide composition and LC-MS/MS analyses did not produce evidence for O glycosylation. 16445295 0 60 from shift 115:119 arg1 point 140:144 the isoelectric point 124:144 the isoelectric point 124:144 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 7 61 attach attached 1108:1115 arg2 homogeneous 1141:1151 homogeneous 1141:1151 homogeneous 1141:1151 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 61 attach attached 1108:1115 arg2 glycan 1101:1106 the glycan 1097:1106 the glycan attached to Asn22 1097:1124 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 7 61 attach attached 1108:1115 arg1 Asn22 1120:1124 Asn22 1120:1124 Asn22 1120:1124 Four of the glycans, Asn51, Asn63, Asn86, and Asn219 displayed microheterogeneity, while the glycan attached to Asn22 appeared to be homogeneous. 16445295 15 62 from propensity 2154:2163 arg1 pI 2145:2146 pI 2145:2146 pI 2145:2146 However, after activation, TAFIa loses 80% of the attached glycans, generating a large shift in pI and a propensity to precipitate. 16445295 15 63 from shift 2136:2140 arg1 pI 2145:2146 pI 2145:2146 pI 2145:2146 However, after activation, TAFIa loses 80% of the attached glycans, generating a large shift in pI and a propensity to precipitate. 16445295 12 64 theme enzymatic 1773:1781 arg1 instability 1783:1793 the enzymatic instability 1769:1793 the enzymatic instability 1769:1793 This pattern is homologous to pancreatic procarboxypeptidase B, and it is therefore unlikely that permutations in the cysteine connectivity are responsible for the enzymatic instability. 16445295 13 65 theme amino 1849:1853 arg1 sequence 1860:1867 the TAFI amino acid sequence 1840:1867 the TAFI amino acid sequence 1840:1867 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 5 66 dep Asn219 914:919 arg1 located 922:928 located 922:928 located 922:928 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 15 67 theme large 2130:2134 arg1 shift 2136:2140 a large shift 2128:2140 a large shift in pI 2128:2146 However, after activation, TAFIa loses 80% of the attached glycans, generating a large shift in pI and a propensity to precipitate. 16445295 4 68 theme disulfide 730:738 arg1 connectivity 740:751 disulfide connectivity 730:751 disulfide connectivity 730:751 The structural bases for these observations were investigated by characterizing all post-translational modifications, including attached glycans and disulfide connectivity. 16445295 4 69 theme post-translational 665:682 arg1 glycans 718:724 attached glycans 709:724 attached glycans 709:724 The structural bases for these observations were investigated by characterizing all post-translational modifications, including attached glycans and disulfide connectivity. 16445295 4 69 theme post-translational 665:682 arg1 connectivity 740:751 disulfide connectivity 730:751 disulfide connectivity 730:751 The structural bases for these observations were investigated by characterizing all post-translational modifications, including attached glycans and disulfide connectivity. 16445295 4 69 theme post-translational 665:682 arg1 modifications 684:696 all post-translational modifications 661:696 all post-translational modifications 661:696 The structural bases for these observations were investigated by characterizing all post-translational modifications, including attached glycans and disulfide connectivity. 16445295 5 70 from peptide 900:906 arg1 located 874:880 located 874:880 located 874:880 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 0 71 dep modifications 19:31 arg1 evidence 94:101 evidence 94:101 evidence for a large shift in the isoelectric point 94:144 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 71 dep modifications 19:31 arg1 solubility 158:167 reduced solubility 150:167 reduced solubility upon activation 150:183 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 0 71 dep modifications 19:31 arg1 modifications 19:31 Post-translational modifications 0:31 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 0:184 Post-translational modifications of human thrombin-activatable fibrinolysis inhibitor (TAFI): evidence for a large shift in the isoelectric point and reduced solubility upon activation. 16445295 6 72 theme unglycosylated 984:997 arg1 variant 999:1005 an unglycosylated variant 981:1005 an unglycosylated variant 981:1005 Asn219 was also found in an unglycosylated variant. 16445295 15 73 theme glycans 2108:2114 arg1 glycans 2108:2114 the attached glycans 2095:2114 the attached glycans 2095:2114 However, after activation, TAFIa loses 80% of the attached glycans, generating a large shift in pI and a propensity to precipitate. 16445295 15 73 theme glycans 2108:2114 arg1 % 2090:2090 80% 2088:2090 80% of the attached glycans 2088:2114 However, after activation, TAFIa loses 80% of the attached glycans, generating a large shift in pI and a propensity to precipitate. 16445295 13 74 theme TAFI 1844:1847 arg1 sequence 1860:1867 the TAFI amino acid sequence 1840:1867 the TAFI amino acid sequence 1840:1867 LC-MS/MS analyses covering more than 90% of the TAFI amino acid sequence revealed no additional modifications. 16445295 5 75 theme N-glycosylation 800:814 arg1 sites 816:820 all five potential N-glycosylation sites 781:820 all five potential N-glycosylation sites 781:820 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 12 76 from permutations 1707:1718 arg1 connectivity 1736:1747 the cysteine connectivity 1723:1747 the cysteine connectivity 1723:1747 This pattern is homologous to pancreatic procarboxypeptidase B, and it is therefore unlikely that permutations in the cysteine connectivity are responsible for the enzymatic instability. 16445295 5 77 from located 922:928 arg1 domain 947:952 the catalytic domain 933:952 the catalytic domain 933:952 The analyses revealed that all five potential N-glycosylation sites were utilized including Asn22, Asn51, Asn63, Asn86 (located in the activation peptide), and Asn219 (located in the catalytic domain). 16445295 1 78 theme pancreatic 252:261 arg1 B 283:283 pancreatic procarboxypeptidase B 252:283 pancreatic procarboxypeptidase B in several ways 252:299 Thrombin-activable fibrinolysis inhibitor (TAFI) is distinct from pancreatic procarboxypeptidase B in several ways. 2737288 1 0 theme human 120:124 arg1 pancreas 126:133 human pancreas 120:133 human pancreas 120:133 Crystalline elastase 1 from human pancreas was digested with trypsin. 2737288 2 1 gly N-glycosylation 201:215 arg2 sites 217:221 the potential N-glycosylation sites 187:221 the potential N-glycosylation sites 187:221 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 2 2 theme amino 276:280 arg1 analysis 287:294 amino acid analysis 276:294 amino acid analysis 276:294 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 1 3 from pancreas 126:133 arg1 elastase 104:111 Crystalline elastase 1 92:113 Crystalline elastase 1 from human pancreas 92:133 Crystalline elastase 1 from human pancreas was digested with trypsin. 2737288 0 4 theme glycosylation 41:53 arg1 elastase 80:87 human pancreatic elastase 1 63:89 human pancreatic elastase 1 63:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 4 theme glycosylation 41:53 arg1 site 55:58 the glycosylation site 37:58 the glycosylation site of human pancreatic elastase 1 37:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 5 theme elastase 80:87 arg1 elastase 80:87 human pancreatic elastase 1 63:89 human pancreatic elastase 1 63:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 5 theme elastase 80:87 arg1 site 55:58 the glycosylation site 37:58 the glycosylation site of human pancreatic elastase 1 37:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 2 6 theme component 325:333 arg1 analysis 335:342 carbohydrate component analysis 312:342 carbohydrate component analysis 312:342 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 2 7 theme acid 282:285 arg1 analysis 287:294 amino acid analysis 276:294 amino acid analysis 276:294 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 2 8 theme potential 191:199 arg1 sites 217:221 the potential N-glycosylation sites 187:221 the potential N-glycosylation sites 187:221 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 0 9 gly glycosylation 41:53 arg1 elastase 80:87 human pancreatic elastase 1 63:89 human pancreatic elastase 1 63:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 9 gly glycosylation 41:53 arg2 site 55:58 the glycosylation site 37:58 the glycosylation site of human pancreatic elastase 1 37:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 9 gly glycosylation 41:53 arg2 elastase 80:87 human pancreatic elastase 1 63:89 human pancreatic elastase 1 63:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 2 10 theme carbohydrate 312:323 arg1 analysis 335:342 carbohydrate component analysis 312:342 carbohydrate component analysis 312:342 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 0 11 theme pancreatic 69:78 arg1 elastase 80:87 human pancreatic elastase 1 63:89 human pancreatic elastase 1 63:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 1 12 theme Crystalline 92:102 arg1 elastase 104:111 Crystalline elastase 1 92:113 Crystalline elastase 1 from human pancreas 92:133 Crystalline elastase 1 from human pancreas was digested with trypsin. 2737288 2 13 contain containing 176:185 arg2 sites 217:221 the potential N-glycosylation sites 187:221 the potential N-glycosylation sites 187:221 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 2 13 contain containing 176:185 arg1 peptides 166:173 Two peptides 162:173 Two peptides 162:173 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 2737288 3 14 theme only 374:377 arg1 Asn-86 379:384 only Asn-86 374:384 only Asn-86 374:384 The results demonstrate that only Asn-86 is glycosylated. 2737288 0 15 theme human 63:67 arg1 elastase 80:87 human pancreatic elastase 1 63:89 human pancreatic elastase 1 63:89 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 16 theme site 55:58 arg1 characterization 17:32 characterization 17:32 characterization 17:32 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 0 16 theme site 55:58 arg1 Localization 0:11 Localization 0:11 Localization 0:11 Localization and characterization of the glycosylation site of human pancreatic elastase 1. 2737288 3 17 gly glycosylated 389:400 arg1 Asn-86 379:384 only Asn-86 374:384 only Asn-86 374:384 The results demonstrate that only Asn-86 is glycosylated. 2737288 2 18 theme N-glycosylation 201:215 arg1 sites 217:221 the potential N-glycosylation sites 187:221 the potential N-glycosylation sites 187:221 Two peptides, containing the potential N-glycosylation sites at Asn-86 and Asn-125, were isolated and analyzed by amino acid analysis, sequencing and carbohydrate component analysis. 12888562 9 0 theme N443D 1746:1750 arg1 mutant 1752:1757 the N443D mutant 1742:1757 the N443D mutant 1742:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 2 1 theme ECV304 321:326 arg1 cells 328:332 human ECV304 cells 315:332 human ECV304 cells 315:332 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 6 2 theme cellular 1192:1199 arg1 trafficking 1201:1211 cellular trafficking 1192:1211 cellular trafficking 1192:1211 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 6 3 dep activity 1179:1186 arg1 the 1175:1177 the 1175:1177 the 1175:1177 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 2 4 with homology 356:363 arg1 members 370:376 members 370:376 members of the E-NTPDase family 370:400 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 4 5 theme 495-amino 779:787 arg1 protein 794:800 only the 495-amino acid protein 770:800 only the 495-amino acid protein 770:800 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 6 6 theme potential 1103:1111 arg1 roles 1113:1117 The potential roles 1099:1117 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase 1099:1230 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 3 7 theme DNA 491:493 arg1 clone 495:499 a genomic DNA clone 481:499 a genomic DNA clone 481:499 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 1 8 theme ecto-nucleoside 225:239 arg1 diphosphohydrolase 254:271 ecto-nucleoside triphosphate diphosphohydrolase 225:271 the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase) 221:290 Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). 12888562 7 9 theme nucleotidase 1354:1365 arg1 activity 1367:1374 nucleotidase activity 1354:1374 nucleotidase activity 1354:1374 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 4 10 theme CHO-K1 755:760 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 7 11 theme devoid 1344:1349 arg1 protein 1325:1331 a protein 1323:1331 a protein completely devoid of nucleotidase activity 1323:1374 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 4 12 theme acid 789:792 arg1 protein 794:800 only the 495-amino acid protein 770:800 only the 495-amino acid protein 770:800 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 9 13 from reduction 1717:1725 arg1 mutant 1752:1757 the N443D mutant 1742:1757 the N443D mutant 1742:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 5 14 theme beta 989:992 arg1 variants 1014:1021 the NTPDase2 beta and -2 gamma splice variants 976:1021 the NTPDase2 beta and -2 gamma splice variants 976:1021 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 6 15 theme residues 1143:1150 arg1 roles 1113:1117 The potential roles 1099:1117 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase 1099:1230 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 4 16 theme NTPDase2 803:810 arg1 alpha 812:816 NTPDase2 alpha 803:816 NTPDase2 alpha 803:816 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 7 17 from beta 1295:1298 arg1 absent 1276:1281 absent 1276:1281 absent 1276:1281 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 9 18 theme activity 1625:1632 arg1 reduction 1717:1725 the large reduction 1707:1725 the large reduction of activity in the N443D mutant 1707:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 18 theme activity 1625:1632 arg1 lack 1599:1602 the lack 1595:1602 the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma 1595:1672 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 18 theme activity 1625:1632 arg1 due 1763:1765 due 1763:1765 due 1763:1765 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 18 theme activity 1625:1632 arg1 mutant 1688:1693 the C399S mutant 1678:1693 the C399S mutant 1678:1693 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 19 from alterations 1770:1780 arg1 folding/maturation 1789:1806 the folding/maturation 1785:1806 the folding/maturation of these proteins 1785:1824 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 0 20 theme variants 140:147 arg1 activities 119:128 the activities 115:128 the activities of splice variants of the enzyme 115:161 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 0 21 theme ecto-ATPase 50:60 arg1 processing 26:35 processing 26:35 processing of the human ecto-ATPase (NTPDase2) 26:71 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 6 22 from roles 1113:1117 arg1 activity 1179:1186 activity 1179:1186 activity 1179:1186 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 6 22 from roles 1113:1117 arg1 trafficking 1201:1211 cellular trafficking 1192:1211 cellular trafficking 1192:1211 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 9 23 from lack 1599:1602 arg1 mutant 1752:1757 the N443D mutant 1742:1757 the N443D mutant 1742:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 5 24 theme splice 1007:1012 arg1 variants 1014:1021 the NTPDase2 beta and -2 gamma splice variants 976:1021 the NTPDase2 beta and -2 gamma splice variants 976:1021 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 5 25 theme NTPDase2 895:902 arg1 alpha 904:908 NTPDase2 alpha 895:908 NTPDase2 alpha 895:908 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 7 26 theme activity 1441:1448 arg1 loss 1433:1436 substantial loss 1421:1436 substantial loss of activity 1421:1448 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 2 27 theme acids 460:464 arg1 proteins 424:431 predicted proteins 414:431 predicted proteins of 495, 472, and 450 amino acids 414:464 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 2 27 theme acids 460:464 arg1 cDNAs 340:344 three cDNAs 334:344 three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids 334:464 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 4 28 theme similar 698:704 arg1 levels 706:711 similar levels 698:711 similar levels 698:711 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 2 29 theme predicted 414:422 arg1 proteins 424:431 predicted proteins 414:431 predicted proteins of 495, 472, and 450 amino acids 414:464 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 2 29 theme predicted 414:422 arg1 cDNAs 340:344 three cDNAs 334:344 three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids 334:464 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 4 30 theme enzyme 652:657 arg1 forms 659:663 all three enzyme forms 642:663 all three enzyme forms 642:663 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 2 31 theme high 351:354 arg1 homology 356:363 high homology 351:363 high homology with members of the E-NTPDase family 351:400 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 1 32 theme triphosphate 241:252 arg1 diphosphohydrolase 254:271 ecto-nucleoside triphosphate diphosphohydrolase 225:271 the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase) 221:290 Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). 12888562 5 33 theme Immunolocalization 850:867 arg1 studies 869:875 Immunolocalization studies 850:875 Immunolocalization studies 850:875 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 2 34 theme E-NTPDase 385:393 arg1 family 395:400 the E-NTPDase family 381:400 the E-NTPDase family 381:400 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 0 35 theme Cys399 15:20 arg1 Requirement 0:10 Requirement 0:10 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) 0:71 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 0 35 theme Cys399 15:20 arg1 implications 81:92 its implications 77:92 its implications for determination of the activities of splice variants of the enzyme 77:161 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 1 36 theme family 273:278 arg1 enzyme 211:216 the type 2 enzyme 200:216 the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase) 200:290 Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). 12888562 7 37 theme substantial 1421:1431 arg1 loss 1433:1436 substantial loss 1421:1436 substantial loss of activity 1421:1448 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 3 38 theme NTPDase2 594:601 arg1 alpha 603:607 NTPDase2 alpha 594:607 NTPDase2 alpha 594:607 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 4 39 theme cells 738:742 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 3 40 theme ecto-ATPase 581:591 arg1 variants 559:566 splice variants 552:566 splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma) 552:630 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 4 41 theme K-1 750:752 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 2 42 theme human 315:319 arg1 cells 328:332 human ECV304 cells 315:332 human ECV304 cells 315:332 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 8 43 gly glycosylated 1500:1511 arg1 Cys399 1463:1468 the Cys399 1459:1468 the Cys399 1459:1468 Neither the Cys399 nor Asn443 mutants were fully glycosylated, and both were retained in the endoplasmic reticulum. 12888562 8 43 gly glycosylated 1500:1511 arg1 mutants 1481:1487 Asn443 mutants 1474:1487 Asn443 mutants 1474:1487 Neither the Cys399 nor Asn443 mutants were fully glycosylated, and both were retained in the endoplasmic reticulum. 12888562 2 44 with cDNAs 340:344 arg1 homology 356:363 high homology 351:363 high homology with members of the E-NTPDase family 351:400 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 9 45 theme activity 1730:1737 arg1 reduction 1717:1725 the large reduction 1707:1725 the large reduction of activity in the N443D mutant 1707:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 45 theme activity 1730:1737 arg1 lack 1599:1602 the lack 1595:1602 the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma 1595:1672 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 45 theme activity 1730:1737 arg1 due 1763:1765 due 1763:1765 due 1763:1765 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 45 theme activity 1730:1737 arg1 mutant 1688:1693 the C399S mutant 1678:1693 the C399S mutant 1678:1693 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 4 46 theme hamster 724:730 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 1 47 theme type 204:207 arg1 enzyme 211:216 the type 2 enzyme 200:216 the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase) 200:290 Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). 12888562 0 48 theme enzyme 156:161 arg1 variants 140:147 splice variants 133:147 splice variants of the enzyme 133:161 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 9 49 from mutant 1688:1693 arg1 mutant 1752:1757 the N443D mutant 1742:1757 the N443D mutant 1742:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 7 50 theme activity 1367:1374 arg1 devoid 1344:1349 devoid 1344:1349 devoid 1344:1349 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 5 51 from forms 1063:1067 arg1 reticulum 1088:1096 the endoplasmic reticulum 1072:1096 the endoplasmic reticulum 1072:1096 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 3 52 theme genomic 483:489 arg1 clone 495:499 a genomic DNA clone 481:499 a genomic DNA clone 481:499 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 5 53 gly glycosylated 1050:1061 arg1 forms 1063:1067 not fully glycosylated forms 1040:1067 not fully glycosylated forms in the endoplasmic reticulum 1040:1096 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 3 54 theme clone 495:499 arg1 Sequencing 467:476 Sequencing 467:476 Sequencing of a genomic DNA clone 467:499 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 0 55 theme activities 119:128 arg1 determination 98:110 determination 98:110 determination of the activities of splice variants of the enzyme 98:161 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 4 56 theme ecto-ATPase 828:838 arg1 activity 840:847 ecto-ATPase activity 828:847 ecto-ATPase activity 828:847 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 7 57 theme NTPDase2 1286:1293 arg1 beta 1295:1298 NTPDase2 beta 1286:1298 NTPDase2 beta 1286:1298 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 9 58 theme proteins 1817:1824 arg1 folding/maturation 1789:1806 the folding/maturation 1785:1806 the folding/maturation of these proteins 1785:1824 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 6 59 theme ecto-ATPase 1220:1230 arg1 activity 1179:1186 activity 1179:1186 activity 1179:1186 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 6 59 theme ecto-ATPase 1220:1230 arg1 trafficking 1201:1211 cellular trafficking 1192:1211 cellular trafficking 1192:1211 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 5 60 theme gamma 1001:1005 arg1 variants 1014:1021 the NTPDase2 beta and -2 gamma splice variants 976:1021 the NTPDase2 beta and -2 gamma splice variants 976:1021 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 9 61 theme ecto-nucleotidase 1607:1623 arg1 activity 1625:1632 ecto-nucleotidase activity 1607:1632 ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma 1607:1672 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 6 62 theme conserved 1133:1141 arg1 residues 1143:1150 two highly conserved residues 1122:1150 two highly conserved residues 1122:1150 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 6 62 theme conserved 1133:1141 arg1 Asn443 1164:1169 Asn443 1164:1169 Asn443 1164:1169 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 6 62 theme conserved 1133:1141 arg1 Cys399 1153:1158 Cys399 1153:1158 Cys399 1153:1158 The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. 12888562 5 63 theme NTPDase2 980:987 arg1 beta 989:992 NTPDase2 beta 980:992 NTPDase2 beta 980:992 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 3 64 theme splice 552:557 arg1 variants 559:566 splice variants 552:566 splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma) 552:630 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 7 65 from gamma 1307:1311 arg1 absent 1276:1281 absent 1276:1281 absent 1276:1281 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 9 66 theme NTPDase2 1647:1654 arg1 beta 1656:1659 NTPDase2 beta 1647:1659 NTPDase2 beta 1647:1659 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 7 67 theme Cys399 1259:1264 arg1 Mutation 1247:1254 Mutation 1247:1254 Mutation 1247:1254 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 5 68 theme glycosylated 1050:1061 arg1 forms 1063:1067 not fully glycosylated forms 1040:1067 not fully glycosylated forms in the endoplasmic reticulum 1040:1096 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 2 69 theme amino 454:458 arg1 acids 460:464 495, 472, and 450 amino acids 436:464 495, 472, and 450 amino acids 436:464 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 5 70 dep fully 1044:1048 arg1 not 1040:1042 not 1040:1042 not 1040:1042 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 1 71 theme diphosphohydrolase 254:271 arg1 family 273:278 the ecto-nucleoside triphosphate diphosphohydrolase family 221:278 the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase) 221:290 Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). 12888562 1 71 theme diphosphohydrolase 254:271 arg1 E-NTPDase 281:289 E-NTPDase 281:289 E-NTPDase 281:289 Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). 12888562 7 72 from absent 1276:1281 arg1 beta 1295:1298 NTPDase2 beta 1286:1298 NTPDase2 beta 1286:1298 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 7 72 from absent 1276:1281 arg1 gamma 1307:1311 -2 gamma 1304:1311 -2 gamma 1304:1311 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 2 73 theme family 395:400 arg1 members 370:376 members 370:376 members of the E-NTPDase family 370:400 We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. 12888562 0 74 theme splice 133:138 arg1 variants 140:147 splice variants 133:147 splice variants of the enzyme 133:161 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 3 75 theme human 575:579 arg1 ecto-ATPase 581:591 the human ecto-ATPase 571:591 the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma) 571:630 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 4 76 theme ovary 732:736 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 0 77 theme human 44:48 arg1 NTPDase2 63:70 NTPDase2 63:70 NTPDase2 63:70 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 0 77 theme human 44:48 arg1 ecto-ATPase 50:60 the human ecto-ATPase 40:60 the human ecto-ATPase (NTPDase2) 40:71 Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. 12888562 5 78 theme plasma 951:956 arg1 membrane 958:965 the plasma membrane 947:965 the plasma membrane 947:965 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 9 79 theme large 1711:1715 arg1 reduction 1717:1725 the large reduction 1707:1725 the large reduction of activity in the N443D mutant 1707:1757 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 8 80 theme Asn443 1474:1479 arg1 mutants 1481:1487 Asn443 mutants 1474:1487 Asn443 mutants 1474:1487 Neither the Cys399 nor Asn443 mutants were fully glycosylated, and both were retained in the endoplasmic reticulum. 12888562 4 81 theme clone 744:748 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 5 82 theme endoplasmic 1076:1086 arg1 reticulum 1088:1096 the endoplasmic reticulum 1072:1096 the endoplasmic reticulum 1072:1096 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 8 83 theme endoplasmic 1544:1554 arg1 reticulum 1556:1564 the endoplasmic reticulum 1540:1564 the endoplasmic reticulum 1540:1564 Neither the Cys399 nor Asn443 mutants were fully glycosylated, and both were retained in the endoplasmic reticulum. 12888562 3 84 dep ecto-ATPase 581:591 arg1 gamma 625:629 -2 gamma 622:629 -2 gamma 622:629 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 3 84 dep ecto-ATPase 581:591 arg1 alpha 603:607 NTPDase2 alpha 594:607 NTPDase2 alpha 594:607 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 3 84 dep ecto-ATPase 581:591 arg1 beta 612:615 -2 beta 609:615 -2 beta 609:615 Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). 12888562 7 85 theme Asn443 1395:1400 arg1 mutation 1383:1390 mutation 1383:1390 mutation of Asn443 to Asp 1383:1407 Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. 12888562 5 86 dep beta 989:992 arg1 the 976:978 the 976:978 the 976:978 Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. 12888562 4 87 theme Chinese 716:722 arg1 cells 763:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Chinese hamster ovary cells clone K-1 (CHO-K1) cells 716:767 Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. 12888562 9 88 theme C399S 1682:1686 arg1 due 1763:1765 due 1763:1765 due 1763:1765 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 12888562 9 88 theme C399S 1682:1686 arg1 mutant 1688:1693 the C399S mutant 1678:1693 the C399S mutant 1678:1693 These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. 15657036 12 0 theme peptide 1469:1475 arg1 photocross-linking 1435:1452 photocross-linking 1435:1452 photocross-linking of a substrate peptide to proline 182 1435:1490 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 12 0 theme peptide 1469:1475 arg1 assays 1424:1429 yeast two-hybrid interaction assays 1395:1429 yeast two-hybrid interaction assays 1395:1429 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 6 1 theme partial 644:650 arg1 trimming 663:670 partial N-terminal trimming 644:670 partial N-terminal trimming 644:670 Secreted FGE shows partial N-terminal trimming up to residue 73 without loosing catalytic activity. 15657036 12 2 theme yeast 1395:1399 arg1 assays 1424:1429 yeast two-hybrid interaction assays 1395:1429 yeast two-hybrid interaction assays 1395:1429 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 7 3 dep residues 819:826 arg1 178-374 828:834 178-374 828:834 178-374 828:834 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 5 4 theme secreted 611:618 arg1 FGE 620:622 secreted FGE 611:622 secreted FGE 611:622 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 13 5 theme species 1651:1657 arg1 sulfatases 1628:1637 all sulfatases 1624:1637 all sulfatases of the same species 1624:1657 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 10 6 theme protease-resistant 1193:1210 arg1 domain 1212:1217 the N-terminal protease-resistant domain 1178:1217 the N-terminal protease-resistant domain 1178:1217 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 12 7 theme C-terminal 1323:1332 arg1 domain 1334:1339 The C-terminal domain 1319:1339 The C-terminal domain 1319:1339 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 12 8 theme interaction 1412:1422 arg1 assays 1424:1429 yeast two-hybrid interaction assays 1395:1429 yeast two-hybrid interaction assays 1395:1429 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 10 9 theme N-terminal 1182:1191 arg1 domain 1212:1217 the N-terminal protease-resistant domain 1178:1217 the N-terminal protease-resistant domain 1178:1217 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 3 10 theme endoplasmic 365:375 arg1 enzyme 396:401 an endoplasmic reticulum-resident enzyme 362:401 an endoplasmic reticulum-resident enzyme that upon overexpression is secreted 362:438 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 3 10 theme endoplasmic 365:375 arg1 enzyme 315:320 The FGly-generating enzyme 295:320 The FGly-generating enzyme (FGE) 295:326 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 12 11 theme two-hybrid 1401:1410 arg1 assays 1424:1429 yeast two-hybrid interaction assays 1395:1429 yeast two-hybrid interaction assays 1395:1429 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 7 12 theme calcium-binding 734:748 arg1 protein 750:756 a calcium-binding protein 732:756 a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 732:861 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 7 12 theme calcium-binding 734:748 arg1 FGE 725:727 FGE 725:727 FGE 725:727 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 8 13 theme sixth 1034:1038 arg1 residue 1049:1055 the sixth cysteine residue 1030:1055 the sixth cysteine residue 1030:1055 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 5 14 contain contains 530:537 arg1 FGE 526:528 Intracellular FGE 512:528 Intracellular FGE 512:528 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 5 14 contain contains 530:537 arg2 N-glycan 559:566 a high mannose type N-glycan 539:566 a high mannose type N-glycan 539:566 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 12 15 theme substrate 1459:1467 arg1 peptide 1469:1475 a substrate peptide 1457:1475 a substrate peptide 1457:1475 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 2 16 theme residue 286:292 arg1 modification 249:260 post-translational modification 230:260 post-translational modification of a conserved cysteine residue 230:292 In eukaryotes, it is generated in the endoplasmic reticulum by post-translational modification of a conserved cysteine residue. 15657036 9 17 theme innermost 1062:1070 arg1 pair 1081:1084 The innermost cysteine pair 1058:1084 The innermost cysteine pair 1058:1084 The innermost cysteine pair is partially reduced. 15657036 13 18 theme known 1519:1523 arg1 sulfatases 1531:1540 all known human sulfatases 1515:1540 all known human sulfatases 1515:1540 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 6 19 theme Secreted 625:632 arg1 FGE 634:636 Secreted FGE 625:636 Secreted FGE 625:636 Secreted FGE shows partial N-terminal trimming up to residue 73 without loosing catalytic activity. 15657036 10 20 from located 1144:1150 arg1 sequence 1159:1166 the sequence 1155:1166 the sequence preceding the N-terminal protease-resistant domain 1155:1217 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 5 21 theme complex 595:601 arg1 type 603:606 the complex type 591:606 the complex type in secreted FGE 591:622 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 5 22 theme high 541:544 arg1 N-glycan 559:566 a high mannose type N-glycan 539:566 a high mannose type N-glycan 539:566 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 13 23 theme human 1525:1529 arg1 sulfatases 1531:1540 all known human sulfatases 1515:1540 all known human sulfatases 1515:1540 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 0 24 theme Molecular 0:8 arg1 characterization 10:25 Molecular characterization 0:25 Molecular characterization of the human Calpha-formylglycine-generating enzyme. 0:78 Molecular characterization of the human Calpha-formylglycine-generating enzyme. 15657036 8 25 theme cysteine 1040:1047 arg1 residue 1049:1055 the sixth cysteine residue 1030:1055 the sixth cysteine residue 1030:1055 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 6 26 theme catalytic 705:713 arg1 activity 715:722 catalytic activity 705:722 catalytic activity 705:722 Secreted FGE shows partial N-terminal trimming up to residue 73 without loosing catalytic activity. 15657036 3 27 theme reticulum-resident 377:394 arg1 enzyme 396:401 an endoplasmic reticulum-resident enzyme 362:401 an endoplasmic reticulum-resident enzyme that upon overexpression is secreted 362:438 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 3 27 theme reticulum-resident 377:394 arg1 enzyme 315:320 The FGly-generating enzyme 295:320 The FGly-generating enzyme (FGE) 295:326 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 10 28 theme first 1112:1116 arg1 residues 1131:1138 The first two cysteine residues 1108:1138 The first two cysteine residues 1108:1138 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 10 28 theme first 1112:1116 arg1 located 1144:1150 located 1144:1150 located 1144:1150 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 1 29 theme catalytic 115:123 arg1 Calpha-formylglycine 80:99 Calpha-formylglycine 80:99 Calpha-formylglycine (FGly) 80:106 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 1 29 theme catalytic 115:123 arg1 residue 125:131 the catalytic residue 111:131 the catalytic residue in the active site of sulfatases 111:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 13 30 theme purified 1567:1574 arg1 FGE 1576:1578 purified FGE 1567:1578 purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species 1567:1657 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 9 31 theme cysteine 1072:1079 arg1 pair 1081:1084 The innermost cysteine pair 1058:1084 The innermost cysteine pair 1058:1084 The innermost cysteine pair is partially reduced. 15657036 7 32 theme C-terminal 807:816 arg1 domain 856:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 3 33 theme FGly-generating 299:313 arg1 FGE 323:325 FGE 323:325 FGE 323:325 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 3 33 theme FGly-generating 299:313 arg1 enzyme 396:401 an endoplasmic reticulum-resident enzyme 362:401 an endoplasmic reticulum-resident enzyme that upon overexpression is secreted 362:438 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 3 33 theme FGly-generating 299:313 arg1 enzyme 315:320 The FGly-generating enzyme 295:320 The FGly-generating enzyme (FGE) 295:326 The FGly-generating enzyme (FGE), performing this modification, is an endoplasmic reticulum-resident enzyme that upon overexpression is secreted. 15657036 13 34 theme same 1646:1649 arg1 species 1651:1657 the same species 1642:1657 the same species 1642:1657 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 7 35 theme protease-resistant 837:854 arg1 domain 856:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 8 36 theme clamp-like 930:939 arg1 manner 941:946 a clamp-like manner 928:946 a clamp-like manner 928:946 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 8 37 theme disulfide 898:906 arg1 bridges 908:914 three disulfide bridges 892:914 three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue 892:1055 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 0 38 theme Calpha-formylglycine-generating 40:70 arg1 enzyme 72:77 the human Calpha-formylglycine-generating enzyme 30:77 the human Calpha-formylglycine-generating enzyme 30:77 Molecular characterization of the human Calpha-formylglycine-generating enzyme. 15657036 8 39 attach links 955:959 arg1 fifth 1021:1025 fifth 1021:1025 fifth 1021:1025 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 8 39 attach links 955:959 arg2 manner 941:946 a clamp-like manner 928:946 a clamp-like manner 928:946 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 8 39 attach links 955:959 arg1 fourth 990:995 fourth 990:995 fourth 990:995 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 7 40 dep N-terminal 772:781 arg1 residues 784:791 residues 86-168 784:798 residues 86-168 784:798 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 12 41 theme binding 1365:1371 arg1 site 1373:1376 the substrate binding site 1351:1376 the substrate binding site 1351:1376 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 5 42 theme Intracellular 512:524 arg1 FGE 526:528 Intracellular FGE 512:528 Intracellular FGE 512:528 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 0 43 theme human 34:38 arg1 enzyme 72:77 the human Calpha-formylglycine-generating enzyme 30:77 the human Calpha-formylglycine-generating enzyme 30:77 Molecular characterization of the human Calpha-formylglycine-generating enzyme. 15657036 10 44 from sequence 1159:1166 arg1 residues 1131:1138 The first two cysteine residues 1108:1138 The first two cysteine residues 1108:1138 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 10 44 from sequence 1159:1166 arg1 located 1144:1150 located 1144:1150 located 1144:1150 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 2 45 theme post-translational 230:247 arg1 modification 249:260 post-translational modification 230:260 post-translational modification of a conserved cysteine residue 230:292 In eukaryotes, it is generated in the endoplasmic reticulum by post-translational modification of a conserved cysteine residue. 15657036 11 46 theme intermolecular 1252:1265 arg1 bonds 1277:1281 intramolecular or intermolecular disulfide bonds 1234:1281 intramolecular or intermolecular disulfide bonds 1234:1281 They can form intramolecular or intermolecular disulfide bonds, the latter stabilizing homodimers. 15657036 11 46 theme intermolecular 1252:1265 arg1 latter 1288:1293 latter 1288:1293 latter 1288:1293 They can form intramolecular or intermolecular disulfide bonds, the latter stabilizing homodimers. 15657036 7 47 dep residues 784:791 arg1 86-168 793:798 86-168 793:798 86-168 793:798 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 7 48 contain containing 758:767 arg1 protein 750:756 a calcium-binding protein 732:756 a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 732:861 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 7 48 contain containing 758:767 arg1 FGE 725:727 FGE 725:727 FGE 725:727 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 7 48 contain containing 758:767 arg2 domain 856:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 10 49 located located 1144:1150 arg1 sequence 1159:1166 the sequence 1155:1166 the sequence preceding the N-terminal protease-resistant domain 1155:1217 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 10 49 located located 1144:1150 arg2 located 1144:1150 located 1144:1150 located 1144:1150 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 10 49 located located 1144:1150 arg2 residues 1131:1138 The first two cysteine residues 1108:1138 The first two cysteine residues 1108:1138 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 5 50 from type 603:606 arg1 FGE 620:622 secreted FGE 611:622 secreted FGE 611:622 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 0 51 theme enzyme 72:77 arg1 characterization 10:25 Molecular characterization 0:25 Molecular characterization of the human Calpha-formylglycine-generating enzyme. 0:78 Molecular characterization of the human Calpha-formylglycine-generating enzyme. 15657036 11 52 theme disulfide 1267:1275 arg1 bonds 1277:1281 intramolecular or intermolecular disulfide bonds 1234:1281 intramolecular or intermolecular disulfide bonds 1234:1281 They can form intramolecular or intermolecular disulfide bonds, the latter stabilizing homodimers. 15657036 11 52 theme disulfide 1267:1275 arg1 latter 1288:1293 latter 1288:1293 latter 1288:1293 They can form intramolecular or intermolecular disulfide bonds, the latter stabilizing homodimers. 15657036 2 53 theme endoplasmic 205:215 arg1 reticulum 217:225 the endoplasmic reticulum 201:225 the endoplasmic reticulum 201:225 In eukaryotes, it is generated in the endoplasmic reticulum by post-translational modification of a conserved cysteine residue. 15657036 10 54 theme cysteine 1122:1129 arg1 residues 1131:1138 The first two cysteine residues 1108:1138 The first two cysteine residues 1108:1138 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 10 54 theme cysteine 1122:1129 arg1 located 1144:1150 located 1144:1150 located 1144:1150 The first two cysteine residues are located in the sequence preceding the N-terminal protease-resistant domain. 15657036 11 55 theme intramolecular 1234:1247 arg1 bonds 1277:1281 intramolecular or intermolecular disulfide bonds 1234:1281 intramolecular or intermolecular disulfide bonds 1234:1281 They can form intramolecular or intermolecular disulfide bonds, the latter stabilizing homodimers. 15657036 11 55 theme intramolecular 1234:1247 arg1 latter 1288:1293 latter 1288:1293 latter 1288:1293 They can form intramolecular or intermolecular disulfide bonds, the latter stabilizing homodimers. 15657036 4 56 theme Recombinant 441:451 arg1 FGE 453:455 Recombinant FGE 441:455 Recombinant FGE 441:455 Recombinant FGE was purified from cells and secretions to homogeneity. 15657036 2 57 theme cysteine 277:284 arg1 residue 286:292 a conserved cysteine residue 265:292 a conserved cysteine residue 265:292 In eukaryotes, it is generated in the endoplasmic reticulum by post-translational modification of a conserved cysteine residue. 15657036 12 58 theme substrate 1355:1363 arg1 site 1373:1376 the substrate binding site 1351:1376 the substrate binding site 1351:1376 The C-terminal domain comprises the substrate binding site, as evidenced by yeast two-hybrid interaction assays and photocross-linking of a substrate peptide to proline 182. 15657036 1 59 theme active 140:145 arg1 sulfatases 155:164 sulfatases 155:164 sulfatases 155:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 1 59 theme active 140:145 arg1 site 147:150 the active site 136:150 the active site of sulfatases 136:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 1 60 from residue 125:131 arg1 sulfatases 155:164 sulfatases 155:164 sulfatases 155:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 1 60 from residue 125:131 arg1 site 147:150 the active site 136:150 the active site of sulfatases 136:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 13 61 attach derived 1502:1508 arg2 substrates 1552:1561 substrates 1552:1561 substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species 1552:1657 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 13 61 attach derived 1502:1508 arg2 Peptides 1493:1500 Peptides 1493:1500 Peptides derived from all known human sulfatases 1493:1540 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 13 61 attach derived 1502:1508 arg1 sulfatases 1531:1540 all known human sulfatases 1515:1540 all known human sulfatases 1515:1540 Peptides derived from all known human sulfatases served as substrates for purified FGE indicating that FGE is sufficient to modify all sulfatases of the same species. 15657036 2 62 theme conserved 267:275 arg1 residue 286:292 a conserved cysteine residue 265:292 a conserved cysteine residue 265:292 In eukaryotes, it is generated in the endoplasmic reticulum by post-translational modification of a conserved cysteine residue. 15657036 5 63 theme mannose 546:552 arg1 N-glycan 559:566 a high mannose type N-glycan 539:566 a high mannose type N-glycan 539:566 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 5 64 theme type 554:557 arg1 N-glycan 559:566 a high mannose type N-glycan 539:566 a high mannose type N-glycan 539:566 Intracellular FGE contains a high mannose type N-glycan, which is processed to the complex type in secreted FGE. 15657036 8 65 dep fourth 990:995 arg1 seventh 1004:1010 seventh 1004:1010 seventh 1004:1010 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 8 65 dep fourth 990:995 arg1 to 997:998 to 997:998 to 997:998 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 8 65 dep fourth 990:995 arg1 the 974:976 the 974:976 the 974:976 The latter is stabilized by three disulfide bridges arranged in a clamp-like manner, which links the third to the eighth, the fourth to the seventh, and the fifth to the sixth cysteine residue. 15657036 7 66 dep C-terminal 807:816 arg1 residues 819:826 residues 178-374 819:834 residues 178-374 819:834 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 15657036 6 67 dep residue 678:684 arg1 up 672:673 up 672:673 up 672:673 Secreted FGE shows partial N-terminal trimming up to residue 73 without loosing catalytic activity. 15657036 1 68 theme sulfatases 155:164 arg1 sulfatases 155:164 sulfatases 155:164 sulfatases 155:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 1 68 theme sulfatases 155:164 arg1 site 147:150 the active site 136:150 the active site of sulfatases 136:164 Calpha-formylglycine (FGly) is the catalytic residue in the active site of sulfatases. 15657036 6 69 theme N-terminal 652:661 arg1 trimming 663:670 partial N-terminal trimming 644:670 partial N-terminal trimming 644:670 Secreted FGE shows partial N-terminal trimming up to residue 73 without loosing catalytic activity. 15657036 7 70 theme N-terminal 772:781 arg1 domain 856:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain 769:861 FGE is a calcium-binding protein containing an N-terminal (residues 86-168) and a C-terminal (residues 178-374) protease-resistant domain. 16650853 6 0 theme chimeric 1163:1170 arg1 enzyme 1172:1177 A chimeric enzyme 1161:1177 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 1161:1263 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 3 1 theme hydrolyzed 582:591 arg1 UDP-GalNAc 609:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 0 2 theme polypeptide 72:82 arg1 pp-GalNAc-T10 125:137 pp-GalNAc-T10 125:137 pp-GalNAc-T10 125:137 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 0 2 theme polypeptide 72:82 arg1 alpha-N-acetylgalactosaminyltransferase 84:122 polypeptide alpha-N-acetylgalactosaminyltransferase 72:122 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 0:139 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 6 3 theme O-glycans 1451:1459 arg1 range 1442:1446 a wide range 1435:1446 a wide range of O-glycans 1435:1459 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 5 4 theme carbohydrate-binding 1014:1033 arg1 site 1035:1038 the carbohydrate-binding site 1010:1038 the carbohydrate-binding site on the lectin beta sub-domain 1010:1068 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 3 5 theme donor 593:597 arg1 UDP-GalNAc 609:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 5 6 theme peptide 1141:1147 arg1 substrates 1149:1158 GalNAc-glycosylated peptide substrates 1121:1158 GalNAc-glycosylated peptide substrates 1121:1158 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 4 7 theme acceptor 943:950 arg1 substrates 952:961 acceptor substrates 943:961 acceptor substrates 943:961 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 4 8 theme interdomain 853:863 arg1 arrangement 865:875 a distinct interdomain arrangement 842:875 a distinct interdomain arrangement between the catalytic and lectin domains 842:916 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 5 9 theme GalNAc 1097:1102 arg1 glycosylation 1104:1116 GalNAc glycosylation 1097:1116 GalNAc glycosylation 1097:1116 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 6 10 theme various 1371:1377 arg1 specificities 1388:1400 the various acceptor specificities 1367:1400 the various acceptor specificities in different isozymes 1367:1422 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 4 11 theme substrate 814:822 arg1 binding 824:830 donor substrate binding 808:830 donor substrate binding 808:830 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 4 12 theme uncomplexed 684:694 arg1 pp-GalNAc-T1 696:707 uncomplexed pp-GalNAc-T1 684:707 uncomplexed pp-GalNAc-T1 684:707 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 6 13 theme wide 1437:1440 arg1 range 1442:1446 a wide range 1435:1446 a wide range of O-glycans 1435:1459 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 5 14 gly GalNAc-glycosylated 1121:1139 arg1 substrates 1149:1158 GalNAc-glycosylated peptide substrates 1121:1158 GalNAc-glycosylated peptide substrates 1121:1158 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 5 15 theme glycosylation 1104:1116 arg1 position 1085:1092 the position 1081:1092 the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates 1081:1158 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 4 16 theme catalytic 786:794 arg1 center 796:801 the catalytic center 782:801 the catalytic center upon donor substrate binding 782:830 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 2 17 dep transfer 303:310 arg1 alpha-N-acetylgalactosaminyltransferases 391:430 polypeptide alpha-N-acetylgalactosaminyltransferases 379:430 the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts) 299:445 Biosynthesis of the O-glycan is initiated by the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts). 16650853 2 17 dep transfer 303:310 arg1 pp-GalNAc-Ts 433:444 pp-GalNAc-Ts 433:444 pp-GalNAc-Ts 433:444 Biosynthesis of the O-glycan is initiated by the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts). 16650853 0 18 theme activity 42:49 arg1 basis 11:15 Structural basis 0:15 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 0:139 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 4 19 theme catalytic 889:897 arg1 domains 910:916 the catalytic and lectin domains 885:916 domains 910:916 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 3 20 theme crystal 464:470 arg1 structures 472:481 crystal structures 464:481 crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides 464:559 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 4 21 theme distinct 844:851 arg1 arrangement 865:875 a distinct interdomain arrangement 842:875 a distinct interdomain arrangement between the catalytic and lectin domains 842:916 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 6 22 theme pp-GalNAc-T10 1207:1219 arg1 pp-GalNAc-T10 1207:1219 pp-GalNAc-T10 1207:1219 pp-GalNAc-T10 1207:1219 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 6 22 theme pp-GalNAc-T10 1207:1219 arg1 domains 1196:1202 the two domains 1188:1202 the two domains of pp-GalNAc-T10 1188:1219 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 1 23 theme malignant 228:236 arg1 transformation 238:251 malignant transformation 228:251 malignant transformation 228:251 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 0 24 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 0:139 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 0 25 theme carbohydrate 20:31 arg1 activity 42:49 carbohydrate transfer activity 20:49 carbohydrate transfer activity 20:49 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 5 26 theme beta 1054:1057 arg1 sub-domain 1059:1068 the lectin beta sub-domain 1043:1068 the lectin beta sub-domain 1043:1068 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 3 27 with complex 565:571 arg1 UDP-GalNAc 609:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 4 28 theme structural 657:666 arg1 comparison 668:677 A structural comparison 655:677 A structural comparison with uncomplexed pp-GalNAc-T1 655:707 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 6 29 theme potential 1332:1340 arg1 mechanism 1342:1350 a potential mechanism 1330:1350 a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans 1330:1459 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 4 30 theme donor 808:812 arg1 binding 824:830 donor substrate binding 808:830 donor substrate binding 808:830 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 1 31 theme carbohydrate 176:187 arg1 chains 189:194 important carbohydrate chains 166:194 important carbohydrate chains involved in differentiation and malignant transformation 166:251 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 1 31 theme carbohydrate 176:187 arg1 O-glycans 152:160 Mucin-type O-glycans 141:160 Mucin-type O-glycans 141:160 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 3 32 theme pp-GalNAc-T10 490:502 arg1 isozyme 504:510 the pp-GalNAc-T10 isozyme 486:510 the pp-GalNAc-T10 isozyme 486:510 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 0 33 theme transfer 33:40 arg1 activity 42:49 carbohydrate transfer activity 20:49 carbohydrate transfer activity 20:49 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 3 34 theme isozyme 504:510 arg1 structures 472:481 crystal structures 464:481 crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides 464:559 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 4 35 theme narrow 926:931 arg1 cleft 933:937 a narrow cleft 924:937 a narrow cleft for acceptor substrates 924:961 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 3 36 theme glycosylated 539:550 arg1 peptides 552:559 glycosylated peptides 539:559 glycosylated peptides 539:559 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 0 37 theme human 54:58 arg1 UDP-GalNAc 60:69 human UDP-GalNAc 54:69 human UDP-GalNAc 54:69 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 5 38 theme catalytic 989:997 arg1 center 999:1004 the catalytic center 985:1004 the catalytic center 985:1004 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 3 39 gly glycosylated 539:550 arg1 peptides 552:559 glycosylated peptides 539:559 glycosylated peptides 539:559 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 6 40 from pp-GalNAc-T1 1252:1263 arg1 linker 1240:1245 a linker 1238:1245 a linker from pp-GalNAc-T1 1238:1263 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 1 41 theme Mucin-type 141:150 arg1 chains 189:194 important carbohydrate chains 166:194 important carbohydrate chains involved in differentiation and malignant transformation 166:251 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 1 41 theme Mucin-type 141:150 arg1 O-glycans 152:160 Mucin-type O-glycans 141:160 Mucin-type O-glycans 141:160 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 6 42 theme different 1405:1413 arg1 isozymes 1415:1422 different isozymes 1405:1422 different isozymes 1405:1422 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 2 43 theme O-glycan 274:281 arg1 Biosynthesis 254:265 Biosynthesis 254:265 Biosynthesis of the O-glycan 254:281 Biosynthesis of the O-glycan is initiated by the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts). 16650853 6 44 gly non-glycosylated 1290:1305 arg1 acceptors 1307:1315 non-glycosylated acceptors 1290:1315 non-glycosylated acceptors 1290:1315 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 5 45 theme GalNAc-glycosylated 1121:1139 arg1 substrates 1149:1158 GalNAc-glycosylated peptide substrates 1121:1158 GalNAc-glycosylated peptide substrates 1121:1158 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 2 46 theme N-acetylgalactosamine 315:335 arg1 transfer 303:310 the transfer 299:310 the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts) 299:445 Biosynthesis of the O-glycan is initiated by the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts). 16650853 5 47 from site 1035:1038 arg1 sub-domain 1059:1068 the lectin beta sub-domain 1043:1068 the lectin beta sub-domain 1043:1068 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 2 48 theme polypeptide 379:389 arg1 alpha-N-acetylgalactosaminyltransferases 391:430 polypeptide alpha-N-acetylgalactosaminyltransferases 379:430 the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts) 299:445 Biosynthesis of the O-glycan is initiated by the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts). 16650853 2 48 theme polypeptide 379:389 arg1 pp-GalNAc-Ts 433:444 pp-GalNAc-Ts 433:444 pp-GalNAc-Ts 433:444 Biosynthesis of the O-glycan is initiated by the transfer of N-acetylgalactosamine (GalNAc) which is catalyzed by UDP-GalNAc:polypeptide alpha-N-acetylgalactosaminyltransferases (pp-GalNAc-Ts). 16650853 5 49 from position 1085:1092 arg1 substrates 1149:1158 GalNAc-glycosylated peptide substrates 1121:1158 GalNAc-glycosylated peptide substrates 1121:1158 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 3 50 attach present 456:462 arg1 complex 627:633 complex 627:633 complex with GalNAc-serine 627:652 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 3 50 attach present 456:462 arg2 we 453:454 we 453:454 we 453:454 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 3 50 attach present 456:462 arg1 complex 565:571 complex 565:571 complex with the hydrolyzed donor substrate UDP-GalNAc 565:618 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 4 51 theme lectin 903:908 arg1 domains 910:916 the catalytic and lectin domains 885:916 domains 910:916 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 1 52 theme important 166:174 arg1 chains 189:194 important carbohydrate chains 166:194 important carbohydrate chains involved in differentiation and malignant transformation 166:251 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 1 52 theme important 166:174 arg1 O-glycans 152:160 Mucin-type O-glycans 141:160 Mucin-type O-glycans 141:160 Mucin-type O-glycans are important carbohydrate chains involved in differentiation and malignant transformation. 16650853 5 53 theme lectin 1047:1052 arg1 sub-domain 1059:1068 the lectin beta sub-domain 1043:1068 the lectin beta sub-domain 1043:1068 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 3 54 contain has 519:521 arg2 specificity 523:533 specificity 523:533 specificity for glycosylated peptides 523:559 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 3 54 contain has 519:521 arg1 isozyme 504:510 the pp-GalNAc-T10 isozyme 486:510 the pp-GalNAc-T10 isozyme 486:510 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 0 55 dep basis 11:15 arg1 pp-GalNAc-T10 125:137 pp-GalNAc-T10 125:137 pp-GalNAc-T10 125:137 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 0 55 dep basis 11:15 arg1 alpha-N-acetylgalactosaminyltransferase 84:122 polypeptide alpha-N-acetylgalactosaminyltransferase 72:122 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 0:139 Structural basis of carbohydrate transfer activity by human UDP-GalNAc: polypeptide alpha-N-acetylgalactosaminyltransferase (pp-GalNAc-T10). 16650853 5 56 from center 999:1004 arg1 sub-domain 1059:1068 the lectin beta sub-domain 1043:1068 the lectin beta sub-domain 1043:1068 The distance between the catalytic center and the carbohydrate-binding site on the lectin beta sub-domain influences the position of GalNAc glycosylation on GalNAc-glycosylated peptide substrates. 16650853 3 57 theme substrate 599:607 arg1 UDP-GalNAc 609:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 the hydrolyzed donor substrate UDP-GalNAc 578:618 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 6 58 from specificities 1388:1400 arg1 isozymes 1415:1422 different isozymes 1405:1422 different isozymes 1405:1422 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 4 59 theme conformational 735:748 arg1 changes 750:756 substantial conformational changes 723:756 substantial conformational changes 723:756 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 3 60 with complex 627:633 arg1 GalNAc-serine 640:652 GalNAc-serine 640:652 GalNAc-serine 640:652 Here we present crystal structures of the pp-GalNAc-T10 isozyme, which has specificity for glycosylated peptides, in complex with the hydrolyzed donor substrate UDP-GalNAc and in complex with GalNAc-serine. 16650853 6 61 theme non-glycosylated 1290:1305 arg1 acceptors 1307:1315 non-glycosylated acceptors 1290:1315 non-glycosylated acceptors 1290:1315 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 4 62 with comparison 668:677 arg1 pp-GalNAc-T1 696:707 uncomplexed pp-GalNAc-T1 684:707 uncomplexed pp-GalNAc-T1 684:707 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 16650853 6 63 theme acceptor 1379:1386 arg1 specificities 1388:1400 the various acceptor specificities 1367:1400 the various acceptor specificities in different isozymes 1367:1422 A chimeric enzyme in which the two domains of pp-GalNAc-T10 are connected by a linker from pp-GalNAc-T1 acquires activity toward non-glycosylated acceptors, identifying a potential mechanism for generating the various acceptor specificities in different isozymes to produce a wide range of O-glycans. 16650853 4 64 theme substantial 723:733 arg1 changes 750:756 substantial conformational changes 723:756 substantial conformational changes 723:756 A structural comparison with uncomplexed pp-GalNAc-T1 suggests that substantial conformational changes occur in two loops near the catalytic center upon donor substrate binding, and that a distinct interdomain arrangement between the catalytic and lectin domains forms a narrow cleft for acceptor substrates. 23999306 0 0 theme glycans 83:89 arg1 mode 67:70 the binding mode 55:70 the binding mode of complex glycans 55:89 The structure of human α-2,6-sialyltransferase reveals the binding mode of complex glycans. 23999306 4 1 theme cytidine 741:748 arg1 5'-monophosphate 750:765 the product cytidine 5'-monophosphate 729:765 the product cytidine 5'-monophosphate 729:765 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 1 2 theme final 165:169 arg1 pattern 185:191 the final glycosylation pattern 161:191 the final glycosylation pattern of many glycoproteins 161:213 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 10 3 theme configuration 1545:1557 arg1 inversion 1532:1540 inversion 1532:1540 inversion of configuration at the sialyl residue 1532:1579 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 0 4 theme complex 75:81 arg1 glycans 83:89 complex glycans 75:89 complex glycans 75:89 The structure of human α-2,6-sialyltransferase reveals the binding mode of complex glycans. 23999306 6 5 theme fold 990:993 arg1 variant 943:949 a variant 941:949 a variant of the canonical glycosyltransferase A fold 941:993 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 8 6 theme ST6Gal-I 1352:1359 arg1 substrate 1339:1347 a substrate 1337:1347 a substrate of ST6Gal-I 1337:1359 The glycan binding mode can be generalized to any glycoprotein that is a substrate of ST6Gal-I. 23999306 8 6 theme ST6Gal-I 1352:1359 arg1 glycoprotein 1316:1327 any glycoprotein 1312:1327 any glycoprotein that is a substrate of ST6Gal-I 1312:1359 The glycan binding mode can be generalized to any glycoprotein that is a substrate of ST6Gal-I. 23999306 1 7 theme glycosylation 171:183 arg1 pattern 185:191 the final glycosylation pattern 161:191 the final glycosylation pattern of many glycoproteins 161:213 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 7 8 theme ST6Gal-I 1212:1219 arg1 ST6Gal-I 1212:1219 ST6Gal-I 1212:1219 ST6Gal-I 1212:1219 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 7 8 theme ST6Gal-I 1212:1219 arg1 site 1204:1207 the active site 1193:1207 the active site of ST6Gal-I in an orientation compatible with catalysis 1193:1263 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 6 9 theme A 988:988 arg1 fold 990:993 the canonical glycosyltransferase A fold 954:993 the canonical glycosyltransferase A fold 954:993 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 1 10 gly glycoproteins 201:213 arg1 glycoproteins 201:213 many glycoproteins 196:213 many glycoproteins 196:213 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 3 11 theme ST6Gal-I 603:610 arg1 properties 589:598 the enzymatic properties 575:598 the enzymatic properties of ST6Gal-I for biotechnological purposes 575:640 No structural information on ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes is currently available. 23999306 6 12 theme glycosyltransferase 968:986 arg1 fold 990:993 the canonical glycosyltransferase A fold 954:993 the canonical glycosyltransferase A fold 954:993 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 3 13 theme enzymatic 579:587 arg1 properties 589:598 the enzymatic properties 575:598 the enzymatic properties of ST6Gal-I for biotechnological purposes 575:640 No structural information on ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes is currently available. 23999306 10 14 from residue 1573:1579 arg1 inversion 1532:1540 inversion 1532:1540 inversion of configuration at the sialyl residue 1532:1579 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 2 15 theme immunoglobulins 308:322 arg1 sialylation 281:291 Complete sialylation 272:291 Complete sialylation of therapeutic immunoglobulins 272:322 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 7 16 from mate 1175:1178 arg1 glycan 1152:1157 A large glycan 1144:1157 A large glycan from a symmetry mate 1144:1178 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 4 17 theme product 733:739 arg1 5'-monophosphate 750:765 the product cytidine 5'-monophosphate 729:765 the product cytidine 5'-monophosphate 729:765 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 6 18 theme canonical 958:966 arg1 fold 990:993 the canonical glycosyltransferase A fold 954:993 the canonical glycosyltransferase A fold 954:993 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 8 19 theme glycan 1270:1275 arg1 mode 1285:1288 The glycan binding mode 1266:1288 The glycan binding mode 1266:1288 The glycan binding mode can be generalized to any glycoprotein that is a substrate of ST6Gal-I. 23999306 2 20 theme therapeutic 296:306 arg1 immunoglobulins 308:322 therapeutic immunoglobulins 296:322 therapeutic immunoglobulins 296:322 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 9 21 with Comparison 1362:1371 arg1 donor 1442:1446 a modified sialyl donor 1424:1446 a modified sialyl donor 1424:1446 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 9 21 with Comparison 1362:1371 arg1 sialyltransferase 1390:1406 a bacterial sialyltransferase 1378:1406 a bacterial sialyltransferase in complex 1378:1417 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 4 22 theme ST6Gal-I 704:711 arg1 structures 684:693 the crystal structures 672:693 the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate 672:808 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 7 23 theme active 1197:1202 arg1 ST6Gal-I 1212:1219 ST6Gal-I 1212:1219 ST6Gal-I 1212:1219 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 7 23 theme active 1197:1202 arg1 site 1204:1207 the active site 1193:1207 the active site of ST6Gal-I in an orientation compatible with catalysis 1193:1263 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 2 24 gly sialylation 281:291 arg1 protein 378:384 protein stability 378:394 protein stability 378:394 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 2 24 gly sialylation 281:291 arg1 immunoglobulins 308:322 therapeutic immunoglobulins 296:322 therapeutic immunoglobulins 296:322 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 10 25 contain bears 1657:1661 arg2 similarity 1676:1685 a conceptual similarity 1663:1685 a conceptual similarity to serine proteases 1663:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 10 25 contain bears 1657:1661 arg1 mechanism 1642:1650 substrate-assisted catalysis with a charge-relay mechanism 1593:1650 substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases 1593:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 4 26 theme human 698:702 arg1 ST6Gal-I 704:711 human ST6Gal-I 698:711 human ST6Gal-I 698:711 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 8 27 theme binding 1277:1283 arg1 mode 1285:1288 The glycan binding mode 1266:1288 The glycan binding mode 1266:1288 The glycan binding mode can be generalized to any glycoprotein that is a substrate of ST6Gal-I. 23999306 10 28 theme serine 1690:1695 arg1 proteases 1697:1705 serine proteases 1690:1705 serine proteases 1690:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 10 29 theme SN2 1513:1515 arg1 mechanism 1517:1525 an SN2 mechanism 1510:1525 an SN2 mechanism with inversion of configuration at the sialyl residue 1510:1579 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 2 30 theme anti-inflammatory 347:363 arg1 activity 365:372 their anti-inflammatory activity 341:372 their anti-inflammatory activity 341:372 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 10 31 theme charge-relay 1629:1640 arg1 mechanism 1642:1650 substrate-assisted catalysis with a charge-relay mechanism 1593:1650 substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases 1593:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 1 32 theme many 196:199 arg1 glycoproteins 201:213 many glycoproteins 196:213 many glycoproteins 196:213 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 2 33 theme ST6Gal-I 467:474 arg1 activity 455:462 the limited activity 443:462 the limited activity of ST6Gal-I towards some galactose acceptors 443:507 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 9 34 theme modified 1426:1433 arg1 donor 1442:1446 a modified sialyl donor 1424:1446 a modified sialyl donor 1424:1446 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 3 35 from information 524:534 arg1 ST6Gal-I 539:546 ST6Gal-I 539:546 ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes 539:640 No structural information on ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes is currently available. 23999306 1 36 theme Human 92:96 arg1 ST6Gal-I 139:146 ST6Gal-I 139:146 ST6Gal-I 139:146 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 1 36 theme Human 92:96 arg1 I 136:136 Human β-galactoside α-2,6-sialyltransferase I 92:136 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) 92:147 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 1 37 theme glycoproteins 201:213 arg1 pattern 185:191 the final glycosylation pattern 161:191 the final glycosylation pattern of many glycoproteins 161:213 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 0 38 theme human 17:21 arg1 α-2,6-sialyltransferase 23:45 human α-2,6-sialyltransferase 17:45 human α-2,6-sialyltransferase 17:45 The structure of human α-2,6-sialyltransferase reveals the binding mode of complex glycans. 23999306 9 39 theme sialyl 1435:1440 arg1 donor 1442:1446 a modified sialyl donor 1424:1446 a modified sialyl donor 1424:1446 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 7 40 theme symmetry 1166:1173 arg1 mate 1175:1178 a symmetry mate 1164:1178 a symmetry mate 1164:1178 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 8 41 gly glycoprotein 1316:1327 arg1 substrate 1339:1347 a substrate 1337:1347 a substrate of ST6Gal-I 1337:1359 The glycan binding mode can be generalized to any glycoprotein that is a substrate of ST6Gal-I. 23999306 8 41 gly glycoprotein 1316:1327 arg1 glycoprotein 1316:1327 any glycoprotein 1312:1327 any glycoprotein that is a substrate of ST6Gal-I 1312:1359 The glycan binding mode can be generalized to any glycoprotein that is a substrate of ST6Gal-I. 23999306 1 42 theme β-galactoside 98:110 arg1 ST6Gal-I 139:146 ST6Gal-I 139:146 ST6Gal-I 139:146 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 1 42 theme β-galactoside 98:110 arg1 I 136:136 Human β-galactoside α-2,6-sialyltransferase I 92:136 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) 92:147 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 4 43 from structures 684:693 arg1 complex 716:722 complex 716:722 complex with the product cytidine 5'-monophosphate 716:765 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 4 43 from structures 684:693 arg1 complex 774:780 complex 774:780 complex with cytidine and phosphate 774:808 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 2 44 theme limited 447:453 arg1 activity 455:462 the limited activity 443:462 the limited activity of ST6Gal-I towards some galactose acceptors 443:507 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 5 45 theme activity 885:892 arg1 rationalization 851:865 the rationalization 847:865 the rationalization of the inhibitory activity of cytosine-based nucleotides 847:922 These complexes allow the rationalization of the inhibitory activity of cytosine-based nucleotides. 23999306 6 46 theme large 1050:1054 arg1 insertions 1056:1065 several large insertions 1042:1065 several large insertions 1042:1065 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 7 47 with compatible 1239:1248 arg1 catalysis 1255:1263 catalysis 1255:1263 catalysis 1255:1263 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 4 48 with complex 716:722 arg1 5'-monophosphate 750:765 the product cytidine 5'-monophosphate 729:765 the product cytidine 5'-monophosphate 729:765 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 1 49 theme α-2,6-sialyltransferase 112:134 arg1 ST6Gal-I 139:146 ST6Gal-I 139:146 ST6Gal-I 139:146 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 1 49 theme α-2,6-sialyltransferase 112:134 arg1 I 136:136 Human β-galactoside α-2,6-sialyltransferase I 92:136 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) 92:147 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 5 50 theme cytosine-based 897:910 arg1 nucleotides 912:922 cytosine-based nucleotides 897:922 cytosine-based nucleotides 897:922 These complexes allow the rationalization of the inhibitory activity of cytosine-based nucleotides. 23999306 7 51 theme compatible 1239:1248 arg1 orientation 1227:1237 an orientation 1224:1237 an orientation compatible with catalysis 1224:1263 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 6 52 theme several 1042:1048 arg1 insertions 1056:1065 several large insertions 1042:1065 several large insertions 1042:1065 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 5 53 theme nucleotides 912:922 arg1 activity 885:892 the inhibitory activity 870:892 the inhibitory activity of cytosine-based nucleotides 870:922 These complexes allow the rationalization of the inhibitory activity of cytosine-based nucleotides. 23999306 0 54 theme α-2,6-sialyltransferase 23:45 arg1 structure 4:12 The structure 0:12 The structure of human α-2,6-sialyltransferase 0:45 The structure of human α-2,6-sialyltransferase reveals the binding mode of complex glycans. 23999306 7 55 from site 1204:1207 arg1 orientation 1227:1237 an orientation 1224:1237 an orientation compatible with catalysis 1224:1263 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 9 56 from sialyltransferase 1390:1406 arg1 complex 1411:1417 complex 1411:1417 complex 1411:1417 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 5 57 theme inhibitory 874:883 arg1 activity 885:892 the inhibitory activity 870:892 the inhibitory activity of cytosine-based nucleotides 870:922 These complexes allow the rationalization of the inhibitory activity of cytosine-based nucleotides. 23999306 1 58 theme sialyl 233:238 arg1 moiety 240:245 a sialyl moiety 231:245 a sialyl moiety 231:245 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 0 59 theme binding 59:65 arg1 mode 67:70 the binding mode 55:70 the binding mode of complex glycans 55:89 The structure of human α-2,6-sialyltransferase reveals the binding mode of complex glycans. 23999306 7 60 theme large 1146:1150 arg1 glycan 1152:1157 A large glycan 1144:1157 A large glycan from a symmetry mate 1144:1178 A large glycan from a symmetry mate localizes to the active site of ST6Gal-I in an orientation compatible with catalysis. 23999306 3 61 theme biotechnological 616:631 arg1 purposes 633:640 biotechnological purposes 616:640 biotechnological purposes 616:640 No structural information on ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes is currently available. 23999306 9 62 theme bacterial 1380:1388 arg1 sialyltransferase 1390:1406 a bacterial sialyltransferase 1378:1406 a bacterial sialyltransferase in complex 1378:1417 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 6 63 theme related 1012:1018 arg1 sialyltransferases 1020:1037 related sialyltransferases 1012:1037 related sialyltransferases 1012:1037 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 10 64 theme catalysis 1612:1620 arg1 mechanism 1642:1650 substrate-assisted catalysis with a charge-relay mechanism 1593:1650 substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases 1593:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 10 65 with mechanism 1517:1525 arg1 inversion 1532:1540 inversion 1532:1540 inversion of configuration at the sialyl residue 1532:1579 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 10 66 theme sialyl 1566:1571 arg1 residue 1573:1579 the sialyl residue 1562:1579 the sialyl residue 1562:1579 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 2 67 theme protein 378:384 arg1 stability 386:394 protein stability 378:394 protein stability 378:394 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 9 68 theme Michaelis 1471:1479 arg1 complex 1481:1487 the Michaelis complex 1467:1487 the Michaelis complex 1467:1487 Comparison with a bacterial sialyltransferase in complex with a modified sialyl donor lends insight into the Michaelis complex. 23999306 1 69 gly glycosylation 171:183 arg1 glycoproteins 201:213 many glycoproteins 196:213 many glycoproteins 196:213 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 23999306 10 70 theme with 1622:1625 arg1 mechanism 1642:1650 substrate-assisted catalysis with a charge-relay mechanism 1593:1650 substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases 1593:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 2 71 theme Complete 272:279 arg1 sialylation 281:291 Complete sialylation 272:291 Complete sialylation of therapeutic immunoglobulins 272:322 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 3 72 theme structural 513:522 arg1 information 524:534 No structural information 510:534 No structural information on ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes 510:640 No structural information on ST6Gal-I that could help to improve the enzymatic properties of ST6Gal-I for biotechnological purposes is currently available. 23999306 6 73 theme substrate 1121:1129 arg1 specificity 1131:1141 substrate specificity 1121:1141 substrate specificity 1121:1141 ST6Gal-I adopts a variant of the canonical glycosyltransferase A fold and differs from related sialyltransferases by several large insertions and deletions that determine its regiospecificity and substrate specificity. 23999306 10 74 theme substrate-assisted 1593:1610 arg1 mechanism 1642:1650 substrate-assisted catalysis with a charge-relay mechanism 1593:1650 substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases 1593:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 4 75 theme crystal 676:682 arg1 structures 684:693 the crystal structures 672:693 the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate 672:808 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 10 76 theme conceptual 1665:1674 arg1 similarity 1676:1685 a conceptual similarity 1663:1685 a conceptual similarity to serine proteases 1663:1705 The results support an SN2 mechanism with inversion of configuration at the sialyl residue and suggest substrate-assisted catalysis with a charge-relay mechanism that bears a conceptual similarity to serine proteases. 23999306 2 77 theme galactose 489:497 arg1 acceptors 499:507 some galactose acceptors 484:507 some galactose acceptors 484:507 Complete sialylation of therapeutic immunoglobulins is essential for their anti-inflammatory activity and protein stability, but is difficult to achieve in vitro owing to the limited activity of ST6Gal-I towards some galactose acceptors. 23999306 4 78 with complex 774:780 arg1 phosphate 800:808 phosphate 800:808 phosphate 800:808 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 4 78 with complex 774:780 arg1 cytidine 787:794 cytidine 787:794 cytidine 787:794 Here, the crystal structures of human ST6Gal-I in complex with the product cytidine 5'-monophosphate and in complex with cytidine and phosphate are described. 23999306 1 79 theme terminal 252:259 arg1 galactose 261:269 a terminal galactose 250:269 a terminal galactose 250:269 Human β-galactoside α-2,6-sialyltransferase I (ST6Gal-I) establishes the final glycosylation pattern of many glycoproteins by transferring a sialyl moiety to a terminal galactose. 11741940 4 0 theme several 810:816 arg1 proteins 824:831 several other proteins 810:831 several other proteins 810:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 1 1 theme N-terminal 251:260 arg1 domain 262:267 an N-terminal domain 248:267 an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) 248:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 4 2 theme proteins 824:831 arg1 domains 799:805 homologous domains 788:805 homologous domains of several other proteins 788:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 4 2 theme proteins 824:831 arg1 proteins 824:831 several other proteins 810:831 several other proteins 810:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 7 3 theme cysteine 1345:1352 arg1 deletion 1324:1331 deletion 1324:1331 deletion of a single cysteine from the NTR module 1324:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 12 4 theme unprocessed 2113:2123 arg1 protein 2125:2131 unprocessed protein 2113:2131 unprocessed protein detected 2113:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 11 5 theme predominant 1866:1876 arg1 terminus 1880:1887 The predominant N terminus 1862:1887 The predominant N terminus 1862:1887 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 11 5 theme predominant 1866:1876 arg1 Ser 1893:1895 Ser(31) 1893:1899 Ser(31) 1893:1899 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 1 6 theme homologous 269:278 arg1 domain 262:267 an N-terminal domain 248:267 an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) 248:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 6 7 theme heparin-binding 1105:1119 arg1 sFRP-1 1131:1136 sFRP-1 1131:1136 sFRP-1 1131:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 7 theme heparin-binding 1105:1119 arg1 domain 1121:1126 the C-terminal heparin-binding domain 1090:1126 the C-terminal heparin-binding domain of sFRP-1 1090:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 7 theme heparin-binding 1105:1119 arg1 domain 1153:1158 an NTR-type domain 1141:1158 an NTR-type domain 1141:1158 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 10 8 theme processing 1795:1804 arg1 evidence 1757:1764 evidence 1757:1764 evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1757:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 10 9 from evidence 1757:1764 arg1 C 1824:1824 C 1824:1824 C 1824:1824 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 10 9 from evidence 1757:1764 arg1 N 1818:1818 N 1818:1818 N 1818:1818 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 7 10 theme NTR 1363:1365 arg1 module 1367:1372 the NTR module 1359:1372 the NTR module 1359:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 11 from module 1367:1372 arg1 deletion 1324:1331 deletion 1324:1331 deletion of a single cysteine from the NTR module 1324:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 11 from module 1367:1372 arg1 cysteine 1345:1352 a single cysteine 1336:1352 a single cysteine from the NTR module 1336:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 12 theme due 1317:1319 arg1 disulfide 1307:1315 a disulfide 1305:1315 a disulfide due to deletion of a single cysteine from the NTR module 1305:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 13 theme sFRP-1 1263:1268 arg1 sFRP-1 1263:1268 sFRP-1 1263:1268 sFRP-1 1263:1268 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 13 theme sFRP-1 1263:1268 arg1 one 1271:1273 one 1271:1273 one 1271:1273 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 10 14 theme recombinant 1841:1851 arg1 protein 1853:1859 the recombinant protein 1837:1859 the recombinant protein 1837:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 4 15 theme Fz 691:692 arg1 domain 694:699 the Fz domain 687:699 the Fz domain 687:699 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 3 16 theme recombinant 559:569 arg1 sFRP-1 571:576 recombinant sFRP-1 559:576 recombinant sFRP-1 559:576 In this study, the disulfide linkages of recombinant sFRP-1 were determined. 11741940 1 17 theme Frizzled-related 136:151 arg1 protein-1 153:161 Secreted Frizzled-related protein-1 127:161 Secreted Frizzled-related protein-1 (sFRP-1) 127:170 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 17 theme Frizzled-related 136:151 arg1 protein 183:189 a soluble protein 173:189 a soluble protein that binds to Wnts and modulates Wnt signaling 173:236 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 17 theme Frizzled-related 136:151 arg1 sFRP-1 164:169 sFRP-1 164:169 sFRP-1 164:169 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 6 18 theme NTR-type 1144:1151 arg1 sFRP-1 1131:1136 sFRP-1 1131:1136 sFRP-1 1131:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 18 theme NTR-type 1144:1151 arg1 domain 1121:1126 the C-terminal heparin-binding domain 1090:1126 the C-terminal heparin-binding domain of sFRP-1 1090:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 18 theme NTR-type 1144:1151 arg1 domain 1153:1158 an NTR-type domain 1141:1158 an NTR-type domain 1141:1158 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 5 19 theme domain 880:885 arg1 linkages 848:855 The disulfide linkages 834:855 The disulfide linkages of the heparin-binding domain 834:885 The disulfide linkages of the heparin-binding domain are 11-14, 12-15, and 13-16. 11741940 5 19 theme domain 880:885 arg1 11-14 891:895 11-14 891:895 11-14 891:895 The disulfide linkages of the heparin-binding domain are 11-14, 12-15, and 13-16. 11741940 7 20 from deletion 1324:1331 arg1 module 1367:1372 the NTR module 1359:1372 the NTR module 1359:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 6 21 theme experimental 956:967 arg1 verification 969:980 experimental verification 956:980 experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules 956:1047 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 4 22 theme disulfide 665:673 arg1 linkages 675:682 the five disulfide linkages 656:682 the five disulfide linkages in the Fz domain 656:699 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 4 22 theme disulfide 665:673 arg1 consistent 735:744 consistent 735:744 consistent 735:744 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 1 23 theme Fz 326:327 arg1 Frizzled 316:323 Frizzled 316:323 Frizzled (Fz domain) 316:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 23 theme Fz 326:327 arg1 domain 329:334 Fz domain 326:334 Fz domain 326:334 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 8 24 theme potential 1644:1652 arg1 site 1677:1680 the second potential N-linked glycosylation site 1633:1680 the second potential N-linked glycosylation site 1633:1680 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 24 theme potential 1644:1652 arg1 Asn 1623:1625 Asn 1623:1625 Asn(262) 1623:1630 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 7 25 theme protein 1470:1476 arg1 domain 1456:1461 the Fz domain 1449:1461 the Fz domain of the protein 1449:1476 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 25 theme protein 1470:1476 arg1 protein 1470:1476 the protein 1466:1476 the protein 1466:1476 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 12 26 theme terminal 2054:2061 arg1 acid 2069:2072 the terminal amino acid 2050:2072 the terminal amino acid (Lys(313)) 2050:2083 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 4 27 with consistent 735:744 arg1 pattern 765:771 the disulfide pattern 751:771 the disulfide pattern determined for homologous domains of several other proteins 751:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 0 28 theme Frizzled 89:96 arg1 homology 98:105 Frizzled homology 89:105 Frizzled homology 89:105 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 8 29 theme glycosylation 1663:1675 arg1 site 1677:1680 the second potential N-linked glycosylation site 1633:1680 the second potential N-linked glycosylation site 1633:1680 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 29 theme glycosylation 1663:1675 arg1 Asn 1623:1625 Asn 1623:1625 Asn(262) 1623:1630 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 11 30 theme protein 1932:1938 arg1 amounts 1917:1923 minor amounts 1911:1923 minor amounts of the protein with Asp(41) and Phe(50) as the N termini 1911:1980 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 11 30 theme protein 1932:1938 arg1 protein 1932:1938 the protein 1928:1938 the protein with Asp(41) and Phe(50) as the N termini 1928:1980 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 12 31 theme acid 2069:2072 arg1 removal 2039:2045 removal 2039:2045 removal of the terminal amino acid (Lys(313)) 2039:2083 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 31 theme acid 2069:2072 arg1 event 2029:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 8 32 from moiety 1571:1576 arg1 Asn 1581:1583 Asn 1581:1583 Asn(172) (approximately 2.8 kDa) 1581:1612 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 32 from moiety 1571:1576 arg1 kDa 1609:1611 approximately 2.8 kDa 1591:1611 approximately 2.8 kDa 1591:1611 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 32 from moiety 1571:1576 arg1 172 1585:1587 172 1585:1587 172 1585:1587 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 7 33 theme Fz 1453:1454 arg1 domain 1456:1461 the Fz domain 1449:1461 the Fz domain of the protein 1449:1476 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 33 theme Fz 1453:1454 arg1 protein 1470:1476 the protein 1466:1476 the protein 1466:1476 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 1 34 theme heparin-binding 354:368 arg1 domain 370:375 a C-terminal heparin-binding domain 341:375 a C-terminal heparin-binding domain with weak homology to netrin 341:404 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 0 35 theme Disulfide 0:8 arg1 assignments 15:25 Disulfide bond assignments 0:25 Disulfide bond assignments of secreted Frizzled-related protein-1 0:64 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 6 36 theme disulfide 996:1004 arg1 patterns 1006:1013 the disulfide patterns 992:1013 the disulfide patterns proposed for netrin (NTR) modules 992:1047 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 1 37 theme putative 287:294 arg1 Frizzled 316:323 Frizzled 316:323 Frizzled (Fz domain) 316:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 37 theme putative 287:294 arg1 site 308:311 the putative Wnt-binding site 283:311 the putative Wnt-binding site of Frizzled (Fz domain) 283:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 11 38 theme N 1972:1972 arg1 termini 1974:1980 the N termini 1968:1980 the N termini 1968:1980 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 6 39 theme one 985:987 arg1 verification 969:980 experimental verification 956:980 experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules 956:1047 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 0 40 theme secreted 30:37 arg1 protein-1 56:64 secreted Frizzled-related protein-1 30:64 secreted Frizzled-related protein-1 30:64 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 7 41 theme new 1422:1424 arg1 cysteine 1426:1433 a new cysteine 1420:1433 a new cysteine introduced in the Fz domain of the protein 1420:1476 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 8 42 gly glycosylation 1491:1503 arg2 sites 1505:1509 glycosylation sites 1491:1509 glycosylation sites 1491:1509 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 1 43 with domain 262:267 arg1 homology 387:394 weak homology 382:394 weak homology to netrin 382:404 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 7 44 theme linkage 1232:1238 arg1 patterns 1240:1247 alternate disulfide linkage patterns 1212:1247 alternate disulfide linkage patterns 1212:1247 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 0 45 theme protein-1 56:64 arg1 assignments 15:25 Disulfide bond assignments 0:25 Disulfide bond assignments of secreted Frizzled-related protein-1 0:64 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 1 46 theme Frizzled 316:323 arg1 Frizzled 316:323 Frizzled 316:323 Frizzled (Fz domain) 316:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 46 theme Frizzled 316:323 arg1 site 308:311 the putative Wnt-binding site 283:311 the putative Wnt-binding site of Frizzled (Fz domain) 283:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 7 47 theme alternate 1212:1220 arg1 patterns 1240:1247 alternate disulfide linkage patterns 1212:1247 alternate disulfide linkage patterns 1212:1247 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 1 48 theme Wnt 224:226 arg1 signaling 228:236 Wnt signaling 224:236 Wnt signaling 224:236 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 6 49 theme C-terminal 1094:1103 arg1 sFRP-1 1131:1136 sFRP-1 1131:1136 sFRP-1 1131:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 49 theme C-terminal 1094:1103 arg1 domain 1121:1126 the C-terminal heparin-binding domain 1090:1126 the C-terminal heparin-binding domain of sFRP-1 1090:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 49 theme C-terminal 1094:1103 arg1 domain 1153:1158 an NTR-type domain 1141:1158 an NTR-type domain 1141:1158 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 9 50 theme carbohydrate 1712:1723 arg1 groups 1725:1730 No O-linked carbohydrate groups 1700:1730 No O-linked carbohydrate groups 1700:1730 No O-linked carbohydrate groups were detected. 11741940 12 51 theme protein 2125:2131 arg1 amount 2103:2108 only a trace amount 2090:2108 only a trace amount of unprocessed protein detected 2090:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 51 theme protein 2125:2131 arg1 protein 2125:2131 unprocessed protein 2113:2131 unprocessed protein detected 2113:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 8 52 theme carbohydrate 1558:1569 arg1 moiety 1571:1576 a relatively large carbohydrate moiety 1539:1576 a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa) 1539:1612 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 7 53 theme remaining 1387:1395 arg1 cysteine 1397:1404 the remaining cysteine 1383:1404 the remaining cysteine 1383:1404 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 6 54 theme assignments 935:945 arg1 set 928:930 This latter set 916:930 This latter set of assignments 916:945 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 54 theme assignments 935:945 arg1 assignments 935:945 assignments 935:945 assignments 935:945 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 7 55 theme sFRPs 1191:1195 arg1 subsets 1180:1186 two subsets 1176:1186 two subsets of sFRPs 1176:1195 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 8 56 theme sites 1505:1509 arg1 Analysis 1479:1486 Analysis 1479:1486 Analysis of glycosylation sites 1479:1509 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 4 57 from linkages 675:682 arg1 domain 694:699 the Fz domain 687:699 the Fz domain 687:699 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 6 58 theme netrin 1028:1033 arg1 modules 1041:1047 netrin (NTR) modules 1028:1047 netrin (NTR) modules 1028:1047 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 12 59 theme trace 2097:2101 arg1 amount 2103:2108 only a trace amount 2090:2108 only a trace amount of unprocessed protein detected 2090:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 59 theme trace 2097:2101 arg1 protein 2125:2131 unprocessed protein 2113:2131 unprocessed protein detected 2113:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 4 60 theme other 818:822 arg1 proteins 824:831 several other proteins 810:831 several other proteins 810:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 7 61 with pair 1410:1413 arg1 cysteine 1426:1433 a new cysteine 1420:1433 a new cysteine introduced in the Fz domain of the protein 1420:1476 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 1 62 theme soluble 175:181 arg1 protein-1 153:161 Secreted Frizzled-related protein-1 127:161 Secreted Frizzled-related protein-1 (sFRP-1) 127:170 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 62 theme soluble 175:181 arg1 protein 183:189 a soluble protein 173:189 a soluble protein that binds to Wnts and modulates Wnt signaling 173:236 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 7 63 theme single 1338:1343 arg1 cysteine 1345:1352 a single cysteine 1336:1352 a single cysteine from the NTR module 1336:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 4 64 theme N 644:644 arg1 terminus 646:653 the N terminus 640:653 the N terminus 640:653 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 10 65 theme proteolytic 1783:1793 arg1 processing 1795:1804 heterogeneous proteolytic processing 1769:1804 heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1769:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 3 66 theme disulfide 537:545 arg1 linkages 547:554 the disulfide linkages 533:554 the disulfide linkages of recombinant sFRP-1 533:576 In this study, the disulfide linkages of recombinant sFRP-1 were determined. 11741940 4 67 theme homologous 788:797 arg1 domains 799:805 homologous domains 788:805 homologous domains of several other proteins 788:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 4 67 theme homologous 788:797 arg1 proteins 824:831 several other proteins 810:831 several other proteins 810:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 11 68 theme N 1878:1878 arg1 terminus 1880:1887 The predominant N terminus 1862:1887 The predominant N terminus 1862:1887 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 11 68 theme N 1878:1878 arg1 Ser 1893:1895 Ser(31) 1893:1899 Ser(31) 1893:1899 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 8 69 link N-linked 1654:1661 arg1 site 1677:1680 the second potential N-linked glycosylation site 1633:1680 the second potential N-linked glycosylation site 1633:1680 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 69 link N-linked 1654:1661 arg1 Asn 1623:1625 Asn 1623:1625 Asn(262) 1623:1630 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 1 70 theme weak 382:385 arg1 homology 387:394 weak homology 382:394 weak homology to netrin 382:404 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 3 71 theme sFRP-1 571:576 arg1 linkages 547:554 the disulfide linkages 533:554 the disulfide linkages of recombinant sFRP-1 533:576 In this study, the disulfide linkages of recombinant sFRP-1 were determined. 11741940 7 72 theme disulfide 1307:1315 arg1 loss 1297:1300 the loss 1293:1300 the loss of a disulfide due to deletion of a single cysteine from the NTR module 1293:1372 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 12 73 with removal 2039:2045 arg1 amount 2103:2108 only a trace amount 2090:2108 only a trace amount of unprocessed protein detected 2090:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 73 with removal 2039:2045 arg1 protein 2125:2131 unprocessed protein 2113:2131 unprocessed protein detected 2113:2140 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 2 74 dep Fz 472:473 arg1 the 468:470 the 468:470 the 468:470 Both domains are cysteine-rich, having 10 and 6 cysteines in the Fz and heparin-binding domains, respectively. 11741940 1 75 theme Secreted 127:134 arg1 protein-1 153:161 Secreted Frizzled-related protein-1 127:161 Secreted Frizzled-related protein-1 (sFRP-1) 127:170 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 75 theme Secreted 127:134 arg1 protein 183:189 a soluble protein 173:189 a soluble protein that binds to Wnts and modulates Wnt signaling 173:236 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 75 theme Secreted 127:134 arg1 sFRP-1 164:169 sFRP-1 164:169 sFRP-1 164:169 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 5 76 theme heparin-binding 864:878 arg1 domain 880:885 the heparin-binding domain 860:885 the heparin-binding domain 860:885 The disulfide linkages of the heparin-binding domain are 11-14, 12-15, and 13-16. 11741940 1 77 with domain 370:375 arg1 homology 387:394 weak homology 382:394 weak homology to netrin 382:404 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 11 78 theme minor 1911:1915 arg1 amounts 1917:1923 minor amounts 1911:1923 minor amounts of the protein with Asp(41) and Phe(50) as the N termini 1911:1980 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 11 78 theme minor 1911:1915 arg1 protein 1932:1938 the protein 1928:1938 the protein with Asp(41) and Phe(50) as the N termini 1928:1980 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 12 79 theme C-terminal 2007:2016 arg1 removal 2039:2045 removal 2039:2045 removal of the terminal amino acid (Lys(313)) 2039:2083 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 79 theme C-terminal 2007:2016 arg1 event 2029:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 80 theme amino 2063:2067 arg1 acid 2069:2072 the terminal amino acid 2050:2072 the terminal amino acid (Lys(313)) 2050:2083 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 6 81 theme sFRP-1 1131:1136 arg1 sFRP-1 1131:1136 sFRP-1 1131:1136 sFRP-1 1131:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 81 theme sFRP-1 1131:1136 arg1 domain 1121:1126 the C-terminal heparin-binding domain 1090:1126 the C-terminal heparin-binding domain of sFRP-1 1090:1136 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 81 theme sFRP-1 1131:1136 arg1 domain 1153:1158 an NTR-type domain 1141:1158 an NTR-type domain 1141:1158 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 10 82 theme protein 1853:1859 arg1 C 1824:1824 C 1824:1824 C 1824:1824 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 10 82 theme protein 1853:1859 arg1 N 1818:1818 N 1818:1818 N 1818:1818 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 8 83 dep contains 1530:1537 arg1 whereas 1615:1621 whereas 1615:1621 whereas 1615:1621 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 10 84 from N 1818:1818 arg1 processing 1795:1804 heterogeneous proteolytic processing 1769:1804 heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1769:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 10 84 from N 1818:1818 arg1 evidence 1757:1764 evidence 1757:1764 evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1757:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 8 85 theme second 1637:1642 arg1 site 1677:1680 the second potential N-linked glycosylation site 1633:1680 the second potential N-linked glycosylation site 1633:1680 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 85 theme second 1637:1642 arg1 Asn 1623:1625 Asn 1623:1625 Asn(262) 1623:1630 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 5 86 theme disulfide 838:846 arg1 linkages 848:855 The disulfide linkages 834:855 The disulfide linkages of the heparin-binding domain 834:885 The disulfide linkages of the heparin-binding domain are 11-14, 12-15, and 13-16. 11741940 5 86 theme disulfide 838:846 arg1 11-14 891:895 11-14 891:895 11-14 891:895 The disulfide linkages of the heparin-binding domain are 11-14, 12-15, and 13-16. 11741940 8 87 contain contains 1530:1537 arg1 sFRP-1 1523:1528 sFRP-1 1523:1528 sFRP-1 1523:1528 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 87 contain contains 1530:1537 arg2 moiety 1571:1576 a relatively large carbohydrate moiety 1539:1576 a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa) 1539:1612 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 0 88 theme netrin 111:116 arg1 modules 118:124 netrin modules 111:124 netrin modules 111:124 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 10 89 from C 1824:1824 arg1 processing 1795:1804 heterogeneous proteolytic processing 1769:1804 heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1769:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 10 89 from C 1824:1824 arg1 evidence 1757:1764 evidence 1757:1764 evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1757:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 8 90 theme N-linked 1654:1661 arg1 site 1677:1680 the second potential N-linked glycosylation site 1633:1680 the second potential N-linked glycosylation site 1633:1680 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 90 theme N-linked 1654:1661 arg1 Asn 1623:1625 Asn 1623:1625 Asn(262) 1623:1630 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 6 91 theme latter 921:926 arg1 set 928:930 This latter set 916:930 This latter set of assignments 916:945 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 91 theme latter 921:926 arg1 assignments 935:945 assignments 935:945 assignments 935:945 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 10 92 dep N 1818:1818 arg1 the 1814:1816 the 1814:1816 the 1814:1816 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 10 92 dep N 1818:1818 arg1 termini 1826:1832 termini 1826:1832 termini 1826:1832 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 11741940 12 93 theme major 2001:2005 arg1 removal 2039:2045 removal 2039:2045 removal of the terminal amino acid (Lys(313)) 2039:2083 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 93 theme major 2001:2005 arg1 event 2029:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 1 94 theme C-terminal 343:352 arg1 domain 370:375 a C-terminal heparin-binding domain 341:375 a C-terminal heparin-binding domain with weak homology to netrin 341:404 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 2 95 theme heparin-binding 479:493 arg1 domains 495:501 heparin-binding domains 479:501 heparin-binding domains 479:501 Both domains are cysteine-rich, having 10 and 6 cysteines in the Fz and heparin-binding domains, respectively. 11741940 12 96 theme processing 2018:2027 arg1 removal 2039:2045 removal 2039:2045 removal of the terminal amino acid (Lys(313)) 2039:2083 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 96 theme processing 2018:2027 arg1 event 2029:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event 1997:2033 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 0 97 theme bond 10:13 arg1 assignments 15:25 Disulfide bond assignments 0:25 Disulfide bond assignments of secreted Frizzled-related protein-1 0:64 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 7 98 contain have 1207:1210 arg1 subsets 1180:1186 two subsets 1176:1186 two subsets of sFRPs 1176:1195 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 7 98 contain have 1207:1210 arg2 patterns 1240:1247 alternate disulfide linkage patterns 1212:1247 alternate disulfide linkage patterns 1212:1247 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 6 99 theme patterns 1006:1013 arg1 patterns 1006:1013 the disulfide patterns 992:1013 the disulfide patterns proposed for netrin (NTR) modules 992:1047 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 6 99 theme patterns 1006:1013 arg1 one 985:987 one 985:987 one 985:987 This latter set of assignments provides experimental verification of one of the disulfide patterns proposed for netrin (NTR) modules and thereby supports the prediction that the C-terminal heparin-binding domain of sFRP-1 is an NTR-type domain. 11741940 8 100 gly glycosylation 1663:1675 arg2 Asn 1623:1625 Asn 1623:1625 Asn(262) 1623:1630 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 8 100 gly glycosylation 1663:1675 arg2 site 1677:1680 the second potential N-linked glycosylation site 1633:1680 the second potential N-linked glycosylation site 1633:1680 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 1 101 contain contains 239:246 arg1 protein-1 153:161 Secreted Frizzled-related protein-1 127:161 Secreted Frizzled-related protein-1 (sFRP-1) 127:170 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 101 contain contains 239:246 arg2 domain 370:375 a C-terminal heparin-binding domain 341:375 a C-terminal heparin-binding domain with weak homology to netrin 341:404 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 101 contain contains 239:246 arg2 domain 262:267 an N-terminal domain 248:267 an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) 248:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 101 contain contains 239:246 arg1 protein 183:189 a soluble protein 173:189 a soluble protein that binds to Wnts and modulates Wnt signaling 173:236 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 101 contain contains 239:246 arg1 sFRP-1 164:169 sFRP-1 164:169 sFRP-1 164:169 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 0 102 theme Frizzled-related 39:54 arg1 protein-1 56:64 secreted Frizzled-related protein-1 30:64 secreted Frizzled-related protein-1 30:64 Disulfide bond assignments of secreted Frizzled-related protein-1 provide insights about Frizzled homology and netrin modules. 11741940 1 103 theme Wnt-binding 296:306 arg1 Frizzled 316:323 Frizzled 316:323 Frizzled (Fz domain) 316:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 1 103 theme Wnt-binding 296:306 arg1 site 308:311 the putative Wnt-binding site 283:311 the putative Wnt-binding site of Frizzled (Fz domain) 283:335 Secreted Frizzled-related protein-1 (sFRP-1), a soluble protein that binds to Wnts and modulates Wnt signaling, contains an N-terminal domain homologous to the putative Wnt-binding site of Frizzled (Fz domain) and a C-terminal heparin-binding domain with weak homology to netrin. 11741940 7 104 theme disulfide 1222:1230 arg1 patterns 1240:1247 alternate disulfide linkage patterns 1212:1247 alternate disulfide linkage patterns 1212:1247 Interestingly, two subsets of sFRPs appear to have alternate disulfide linkage patterns compared with sFRP-1, one of which involves the loss of a disulfide due to deletion of a single cysteine from the NTR module, whereas the remaining cysteine may pair with a new cysteine introduced in the Fz domain of the protein. 11741940 12 105 dep acid 2069:2072 arg1 313 2079:2081 313 2079:2081 313 2079:2081 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 12 105 dep acid 2069:2072 arg1 Lys 2075:2077 Lys 2075:2077 Lys 2075:2077 The major C-terminal processing event was removal of the terminal amino acid (Lys(313)) with only a trace amount of unprocessed protein detected. 11741940 4 106 theme disulfide 755:763 arg1 pattern 765:771 the disulfide pattern 751:771 the disulfide pattern determined for homologous domains of several other proteins 751:831 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 8 107 theme large 1552:1556 arg1 moiety 1571:1576 a relatively large carbohydrate moiety 1539:1576 a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa) 1539:1612 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 9 108 theme O-linked 1703:1710 arg1 groups 1725:1730 No O-linked carbohydrate groups 1700:1730 No O-linked carbohydrate groups 1700:1730 No O-linked carbohydrate groups were detected. 11741940 8 109 theme glycosylation 1491:1503 arg1 sites 1505:1509 glycosylation sites 1491:1509 glycosylation sites 1491:1509 Analysis of glycosylation sites showed that sFRP-1 contains a relatively large carbohydrate moiety on Asn(172) (approximately 2.8 kDa), whereas Asn(262), the second potential N-linked glycosylation site, is not modified. 11741940 11 110 with protein 1932:1938 arg1 50 1961:1962 50 1961:1962 50 1961:1962 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 11 110 with protein 1932:1938 arg1 Phe 1957:1959 Phe 1957:1959 Phe 1957:1959 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 11 110 with protein 1932:1938 arg1 Asp 1945:1947 Asp(41) 1945:1951 Asp(41) 1945:1951 The predominant N terminus was Ser(31), although minor amounts of the protein with Asp(41) and Phe(50) as the N termini were observed. 11741940 9 111 link O-linked 1703:1710 arg1 groups 1725:1730 No O-linked carbohydrate groups 1700:1730 No O-linked carbohydrate groups 1700:1730 No O-linked carbohydrate groups were detected. 11741940 4 112 theme sFRP-1 605:610 arg1 cysteines 612:620 sFRP-1 cysteines 605:620 sFRP-1 cysteines 605:620 Numbering sFRP-1 cysteines sequentially from the N terminus, the five disulfide linkages in the Fz domain are 1-5, 2-4, 3-8, 6-10, and 7-9, consistent with the disulfide pattern determined for homologous domains of several other proteins. 11741940 10 113 theme heterogeneous 1769:1781 arg1 processing 1795:1804 heterogeneous proteolytic processing 1769:1804 heterogeneous proteolytic processing at both the N and C termini of the recombinant protein 1769:1859 There was evidence of heterogeneous proteolytic processing at both the N and C termini of the recombinant protein. 18703501 7 0 theme Mass 1161:1164 arg1 spectrometry 1166:1177 Mass spectrometry 1161:1177 Mass spectrometry experiments 1161:1189 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 8 1 theme glycosylation 1520:1532 arg1 Thr 1540:1542 Thr(78) 1540:1546 Thr(78) 1540:1546 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 1 theme glycosylation 1520:1532 arg1 site 1534:1537 an additional O-linked glycosylation site 1497:1537 an additional O-linked glycosylation site (Thr(78)) 1497:1547 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 6 2 from Investigations 985:998 arg1 stability 1027:1035 stability 1027:1035 stability 1027:1035 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 6 2 from Investigations 985:998 arg1 oligomerization 1007:1021 oligomerization 1007:1021 oligomerization 1007:1021 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 7 3 theme mannose-type 1341:1352 arg1 structures 1361:1370 high mannose-type glycan structures 1336:1370 high mannose-type glycan structures 1336:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 9 4 theme unique 1784:1789 arg1 enzyme 1791:1796 this unique enzyme 1779:1796 this unique enzyme 1779:1796 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 3 5 theme catalytic 653:661 arg1 domains 663:669 catalytic domains 653:669 catalytic domains (PG + CA form) 653:684 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 1 6 theme pH 209:210 arg1 regulation 212:221 pH regulation 209:221 pH regulation 209:221 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 8 7 from experiments 1382:1392 arg1 protein 1411:1417 a recombinant protein 1397:1417 a recombinant protein obtained by a mammalian cell expression system 1397:1464 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 8 theme oligosaccharide 1585:1599 arg1 structures 1601:1610 the oligosaccharide structures 1581:1610 the oligosaccharide structures 1581:1610 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 5 9 theme Stopped-flow 779:790 arg1 experiments 810:820 Stopped-flow spectrophotometry experiments 779:820 Stopped-flow spectrophotometry experiments on both proteins 779:837 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 4 10 theme IX 775:776 arg1 regions 761:767 the small transmembrane and intracytoplasmic regions 716:767 the small transmembrane and intracytoplasmic regions of CA IX 716:776 The produced proteins lacked the small transmembrane and intracytoplasmic regions of CA IX. 18703501 7 11 theme glycosylation 1295:1307 arg1 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 11 theme glycosylation 1295:1307 arg1 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 12 theme unique 1279:1284 arg1 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 12 theme unique 1279:1284 arg1 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 1 13 theme protein 156:162 arg1 family 164:169 the CA protein family 149:169 the CA protein family 149:169 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 5 14 theme highest 924:930 arg1 activity 942:949 the highest catalytic activity 920:949 the highest catalytic activity ever measured for any CA isozyme 920:982 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 1 15 from role 201:204 arg1 regulation 212:221 pH regulation 209:221 pH regulation 209:221 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 3 16 theme IX 548:549 arg1 domain 561:566 the CA IX catalytic domain 541:566 the CA IX catalytic domain only (CA form) 541:581 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 3 16 theme IX 548:549 arg1 form 577:580 CA form 574:580 CA form 574:580 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 5 17 theme CA 973:974 arg1 isozyme 976:982 any CA isozyme 969:982 any CA isozyme 969:982 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 1 18 dep role 201:204 arg1 addition 175:182 addition 175:182 addition 175:182 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 6 19 theme intermolecular 1127:1140 arg1 s 1157:1157 s 1157:1157 s 1157:1157 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 6 19 theme intermolecular 1127:1140 arg1 bond 1152:1155 intermolecular disulfide bond 1127:1155 intermolecular disulfide bond(s) 1127:1158 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 0 20 theme active 55:60 arg1 anhydrase 71:79 the most active carbonic anhydrase 46:79 the most active carbonic anhydrase isozymes 46:88 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 1 21 theme cell 268:271 arg1 proliferation 273:285 cell proliferation 268:285 cell proliferation 268:285 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 2 22 theme cell 472:475 arg1 system 488:493 the baculovirus-insect cell expression system 449:493 the baculovirus-insect cell expression system 449:493 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 1 23 theme Carbonic 91:98 arg1 anhydrase 100:108 Carbonic anhydrase IX 91:111 Carbonic anhydrase IX (CA IX) 91:119 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 1 23 theme Carbonic 91:98 arg1 CA 114:115 CA IX 114:118 CA IX 114:118 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 1 23 theme Carbonic 91:98 arg1 member 139:144 an exceptional member 124:144 an exceptional member of the CA protein family 124:169 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 0 24 theme anhydrase 71:79 arg1 isozymes 81:88 the most active carbonic anhydrase isozymes 46:88 the most active carbonic anhydrase isozymes 46:88 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 7 25 contain contains 1209:1216 arg2 bridge 1246:1251 an intramolecular disulfide bridge 1218:1251 an intramolecular disulfide bridge (Cys(119)-Cys(299)) 1218:1271 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 25 contain contains 1209:1216 arg2 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 25 contain contains 1209:1216 arg1 IX 1206:1207 CA IX 1203:1207 CA IX 1203:1207 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 25 contain contains 1209:1216 arg2 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 5 26 theme CA 902:903 arg1 form 905:908 CA form 902:908 CA form 902:908 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 2 27 theme membrane 381:388 arg1 protein 390:396 this membrane protein 376:396 this membrane protein 376:396 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 0 28 theme Biochemical 0:10 arg1 isozymes 81:88 the most active carbonic anhydrase isozymes 46:88 the most active carbonic anhydrase isozymes 46:88 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 0 28 theme Biochemical 0:10 arg1 one 39:41 one 39:41 one 39:41 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 0 28 theme Biochemical 0:10 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization of CA IX 0:36 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 5 29 theme certain 874:880 arg1 ions 888:891 certain metal ions 874:891 certain metal ions 874:891 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 7 30 theme Cys 1254:1256 arg1 299 1267:1269 299 1267:1269 299 1267:1269 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 30 theme Cys 1254:1256 arg1 -Cys 1262:1265 Cys(119)-Cys 1254:1265 Cys(119)-Cys(299) 1254:1270 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 2 31 theme biochemical 350:360 arg1 properties 362:371 the biochemical properties 346:371 the biochemical properties of this membrane protein 346:396 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 5 32 theme ions 888:891 arg1 excess 864:869 the excess 860:869 the excess of certain metal ions 860:891 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 2 33 theme soluble 403:409 arg1 forms 423:427 two soluble recombinant forms 399:427 two soluble recombinant forms 399:427 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 8 34 theme mammalian 1433:1441 arg1 system 1459:1464 a mammalian cell expression system 1431:1464 a mammalian cell expression system 1431:1464 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 4 35 theme transmembrane 726:738 arg1 regions 761:767 the small transmembrane and intracytoplasmic regions 716:767 the small transmembrane and intracytoplasmic regions of CA IX 716:776 The produced proteins lacked the small transmembrane and intracytoplasmic regions of CA IX. 18703501 0 36 theme IX 35:36 arg1 isozymes 81:88 the most active carbonic anhydrase isozymes 46:88 the most active carbonic anhydrase isozymes 46:88 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 0 36 theme IX 35:36 arg1 one 39:41 one 39:41 one 39:41 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 0 36 theme IX 35:36 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization of CA IX 0:36 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 7 37 theme intramolecular 1221:1234 arg1 bridge 1246:1251 an intramolecular disulfide bridge 1218:1251 an intramolecular disulfide bridge (Cys(119)-Cys(299)) 1218:1271 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 8 38 theme expression 1448:1457 arg1 system 1459:1464 a mammalian cell expression system 1431:1464 a mammalian cell expression system 1431:1464 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 9 39 theme novel 1633:1637 arg1 information 1639:1649 novel information 1633:1649 novel information on the biochemical properties of CA IX 1633:1688 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 4 40 theme intracytoplasmic 744:759 arg1 regions 761:767 the small transmembrane and intracytoplasmic regions 716:767 the small transmembrane and intracytoplasmic regions of CA IX 716:776 The produced proteins lacked the small transmembrane and intracytoplasmic regions of CA IX. 18703501 9 41 from information 1639:1649 arg1 properties 1670:1679 the biochemical properties 1654:1679 the biochemical properties of CA IX 1654:1688 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 9 42 theme cellular 1728:1735 arg1 processes 1760:1768 the various cellular and pathophysiological processes 1716:1768 the various cellular and pathophysiological processes in which this unique enzyme is involved 1716:1808 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 4 43 theme small 720:724 arg1 regions 761:767 the small transmembrane and intracytoplasmic regions 716:767 the small transmembrane and intracytoplasmic regions of CA IX 716:776 The produced proteins lacked the small transmembrane and intracytoplasmic regions of CA IX. 18703501 8 44 gly glycosylation 1520:1532 arg2 Thr 1540:1542 Thr(78) 1540:1546 Thr(78) 1540:1546 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 44 gly glycosylation 1520:1532 arg2 site 1534:1537 an additional O-linked glycosylation site 1497:1537 an additional O-linked glycosylation site (Thr(78)) 1497:1547 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 7 45 theme CA 1203:1204 arg1 IX 1206:1207 CA IX 1203:1207 CA IX 1203:1207 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 9 46 theme pathophysiological 1741:1758 arg1 processes 1760:1768 the various cellular and pathophysiological processes 1716:1768 the various cellular and pathophysiological processes in which this unique enzyme is involved 1716:1808 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 3 47 theme extracellular 590:602 arg1 domain 604:609 the extracellular domain 586:609 the extracellular domain 586:609 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 7 48 gly glycosylation 1295:1307 arg2 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 48 gly glycosylation 1295:1307 arg2 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 49 theme glycan 1354:1359 arg1 structures 1361:1370 high mannose-type glycan structures 1336:1370 high mannose-type glycan structures 1336:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 50 dep bridge 1246:1251 arg1 299 1267:1269 299 1267:1269 299 1267:1269 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 50 dep bridge 1246:1251 arg1 -Cys 1262:1265 Cys(119)-Cys 1254:1265 Cys(119)-Cys(299) 1254:1270 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 1 51 theme classical 191:199 arg1 role 201:204 its classical role 187:204 its classical role in pH regulation 187:221 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 7 52 theme high 1336:1339 arg1 structures 1361:1370 high mannose-type glycan structures 1336:1370 high mannose-type glycan structures 1336:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 53 theme spectrometry 1166:1177 arg1 experiments 1179:1189 Mass spectrometry experiments 1161:1189 Mass spectrometry experiments 1161:1189 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 8 54 theme O-linked 1511:1518 arg1 Thr 1540:1542 Thr(78) 1540:1546 Thr(78) 1540:1546 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 54 theme O-linked 1511:1518 arg1 site 1534:1537 an additional O-linked glycosylation site 1497:1537 an additional O-linked glycosylation site (Thr(78)) 1497:1547 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 55 theme site 1534:1537 arg1 occurrence 1483:1492 the occurrence 1479:1492 the occurrence of an additional O-linked glycosylation site (Thr(78)) 1479:1547 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 1 56 theme exceptional 127:137 arg1 anhydrase 100:108 Carbonic anhydrase IX 91:111 Carbonic anhydrase IX (CA IX) 91:119 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 1 56 theme exceptional 127:137 arg1 member 139:144 an exceptional member 124:144 an exceptional member of the CA protein family 124:169 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 9 57 theme various 1720:1726 arg1 processes 1760:1768 the various cellular and pathophysiological processes 1716:1768 the various cellular and pathophysiological processes in which this unique enzyme is involved 1716:1808 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 5 58 theme spectrophotometry 792:808 arg1 experiments 810:820 Stopped-flow spectrophotometry experiments 779:820 Stopped-flow spectrophotometry experiments on both proteins 779:837 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 4 59 theme CA 772:773 arg1 IX 775:776 CA IX 772:776 CA IX 772:776 The produced proteins lacked the small transmembrane and intracytoplasmic regions of CA IX. 18703501 3 60 theme CA 677:678 arg1 form 680:683 CA form 677:683 CA form 677:683 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 3 61 theme recombinant 500:510 arg1 proteins 512:519 The recombinant proteins 496:519 The recombinant proteins 496:519 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 8 62 theme structures 1601:1610 arg1 nature 1571:1576 the nature 1567:1576 the nature of the oligosaccharide structures 1567:1610 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 1 63 theme CA 153:154 arg1 family 164:169 the CA protein family 149:169 the CA protein family 149:169 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 7 64 theme N-linked 1286:1293 arg1 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 64 theme N-linked 1286:1293 arg1 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 1 65 theme tumorigenic 307:317 arg1 processes 319:327 tumorigenic processes 307:327 tumorigenic processes 307:327 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 3 66 theme CA 545:546 arg1 domain 561:566 the CA IX catalytic domain 541:566 the CA IX catalytic domain only (CA form) 541:581 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 3 66 theme CA 545:546 arg1 form 577:580 CA form 574:580 CA form 574:580 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 1 67 theme family 164:169 arg1 anhydrase 100:108 Carbonic anhydrase IX 91:111 Carbonic anhydrase IX (CA IX) 91:119 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 1 67 theme family 164:169 arg1 member 139:144 an exceptional member 124:144 an exceptional member of the CA protein family 124:169 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 2 68 theme expression 477:486 arg1 system 488:493 the baculovirus-insect cell expression system 449:493 the baculovirus-insect cell expression system 449:493 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 7 69 contain bears 1330:1334 arg2 structures 1361:1370 high mannose-type glycan structures 1336:1370 high mannose-type glycan structures 1336:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 69 contain bears 1330:1334 arg1 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 69 contain bears 1330:1334 arg1 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 6 70 theme recombinant 1071:1081 arg1 proteins 1083:1090 both recombinant proteins 1066:1090 both recombinant proteins 1066:1090 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 6 71 theme disulfide 1142:1150 arg1 s 1157:1157 s 1157:1157 s 1157:1157 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 6 71 theme disulfide 1142:1150 arg1 bond 1152:1155 intermolecular disulfide bond 1127:1155 intermolecular disulfide bond(s) 1127:1158 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 3 72 theme catalytic 551:559 arg1 domain 561:566 the CA IX catalytic domain 541:566 the CA IX catalytic domain only (CA form) 541:581 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 3 72 theme catalytic 551:559 arg1 form 577:580 CA form 574:580 CA form 574:580 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 5 73 from experiments 810:820 arg1 proteins 830:837 both proteins 825:837 both proteins 825:837 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 7 74 link N-linked 1286:1293 arg1 Asn 1315:1317 Asn(309) 1315:1322 Asn(309) 1315:1322 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 7 74 link N-linked 1286:1293 arg1 site 1309:1312 a unique N-linked glycosylation site 1277:1312 a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures 1277:1370 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 8 75 theme Parallel 1373:1380 arg1 experiments 1382:1392 Parallel experiments 1373:1392 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system 1373:1464 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 76 link O-linked 1511:1518 arg1 Thr 1540:1542 Thr(78) 1540:1546 Thr(78) 1540:1546 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 76 link O-linked 1511:1518 arg1 site 1534:1537 an additional O-linked glycosylation site 1497:1537 an additional O-linked glycosylation site (Thr(78)) 1497:1547 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 9 77 theme biochemical 1658:1668 arg1 properties 1670:1679 the biochemical properties 1654:1679 the biochemical properties of CA IX 1654:1688 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 0 78 theme carbonic 62:69 arg1 anhydrase 71:79 the most active carbonic anhydrase 46:79 the most active carbonic anhydrase isozymes 46:88 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 2 79 theme recombinant 411:421 arg1 forms 423:427 two soluble recombinant forms 399:427 two soluble recombinant forms 399:427 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 0 80 theme isozymes 81:88 arg1 isozymes 81:88 the most active carbonic anhydrase isozymes 46:88 the most active carbonic anhydrase isozymes 46:88 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 0 80 theme isozymes 81:88 arg1 one 39:41 one 39:41 one 39:41 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 0 80 theme isozymes 81:88 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization of CA IX 0:36 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 2 81 theme baculovirus-insect 453:470 arg1 system 488:493 the baculovirus-insect cell expression system 449:493 the baculovirus-insect cell expression system 449:493 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 8 82 theme recombinant 1399:1409 arg1 protein 1411:1417 a recombinant protein 1397:1417 a recombinant protein obtained by a mammalian cell expression system 1397:1464 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 1 83 theme cell 288:291 arg1 adhesion 293:300 cell adhesion 288:300 cell adhesion 288:300 Carbonic anhydrase IX (CA IX) is an exceptional member of the CA protein family; in addition to its classical role in pH regulation, it has also been proposed to participate in cell proliferation, cell adhesion, and tumorigenic processes. 18703501 9 84 theme IX 1687:1688 arg1 properties 1670:1679 the biochemical properties 1654:1679 the biochemical properties of CA IX 1654:1688 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 18703501 0 85 theme CA 32:33 arg1 IX 35:36 CA IX 32:36 CA IX 32:36 Biochemical characterization of CA IX, one of the most active carbonic anhydrase isozymes. 18703501 5 86 theme metal 882:886 arg1 ions 888:891 certain metal ions 874:891 certain metal ions 874:891 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 7 87 theme disulfide 1236:1244 arg1 bridge 1246:1251 an intramolecular disulfide bridge 1218:1251 an intramolecular disulfide bridge (Cys(119)-Cys(299)) 1218:1271 Mass spectrometry experiments showed that CA IX contains an intramolecular disulfide bridge (Cys(119)-Cys(299)) and a unique N-linked glycosylation site (Asn(309)) that bears high mannose-type glycan structures. 18703501 8 88 theme cell 1443:1446 arg1 system 1459:1464 a mammalian cell expression system 1431:1464 a mammalian cell expression system 1431:1464 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 5 89 theme catalytic 932:940 arg1 activity 942:949 the highest catalytic activity 920:949 the highest catalytic activity ever measured for any CA isozyme 920:982 Stopped-flow spectrophotometry experiments on both proteins demonstrated that in the excess of certain metal ions the PG + CA form exhibited the highest catalytic activity ever measured for any CA isozyme. 18703501 3 90 theme CA 574:575 arg1 domain 561:566 the CA IX catalytic domain 541:566 the CA IX catalytic domain only (CA form) 541:581 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 3 90 theme CA 574:575 arg1 form 577:580 CA form 574:580 CA form 574:580 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 6 91 dep oligomerization 1007:1021 arg1 the 1003:1005 the 1003:1005 the 1003:1005 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 2 92 theme protein 390:396 arg1 properties 362:371 the biochemical properties 346:371 the biochemical properties of this membrane protein 346:396 To characterize the biochemical properties of this membrane protein, two soluble recombinant forms were produced using the baculovirus-insect cell expression system. 18703501 3 93 dep domains 663:669 arg1 PG 672:673 PG 672:673 PG 672:673 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 3 93 dep domains 663:669 arg1 form 680:683 CA form 677:683 CA form 677:683 The recombinant proteins consisted of either the CA IX catalytic domain only (CA form) or the extracellular domain, which included both the proteoglycan and catalytic domains (PG + CA form). 18703501 6 94 theme enzymes 1044:1050 arg1 stability 1027:1035 stability 1027:1035 stability 1027:1035 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 6 94 theme enzymes 1044:1050 arg1 oligomerization 1007:1021 oligomerization 1007:1021 oligomerization 1007:1021 Investigations on the oligomerization and stability of the enzymes revealed that both recombinant proteins form dimers that are stabilized by intermolecular disulfide bond(s). 18703501 4 95 theme produced 691:698 arg1 proteins 700:707 The produced proteins 687:707 The produced proteins 687:707 The produced proteins lacked the small transmembrane and intracytoplasmic regions of CA IX. 18703501 8 96 theme additional 1500:1509 arg1 Thr 1540:1542 Thr(78) 1540:1546 Thr(78) 1540:1546 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 8 96 theme additional 1500:1509 arg1 site 1534:1537 an additional O-linked glycosylation site 1497:1537 an additional O-linked glycosylation site (Thr(78)) 1497:1547 Parallel experiments on a recombinant protein obtained by a mammalian cell expression system demonstrated the occurrence of an additional O-linked glycosylation site (Thr(78)) and characterized the nature of the oligosaccharide structures. 18703501 9 97 theme CA 1684:1685 arg1 IX 1687:1688 CA IX 1684:1688 CA IX 1684:1688 This study provides novel information on the biochemical properties of CA IX and may help characterize the various cellular and pathophysiological processes in which this unique enzyme is involved. 11706042 10 0 from CD14 1726:1729 arg1 expression 1687:1696 the surface expression 1675:1696 the surface expression of at least three proteins 1675:1723 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 1 from MD-2 1732:1735 arg1 expression 1687:1696 the surface expression 1675:1696 the surface expression of at least three proteins 1675:1723 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 7 2 theme c-Jun 1317:1321 arg1 JNK 1331:1333 JNK 1331:1333 JNK 1331:1333 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 7 2 theme c-Jun 1317:1321 arg1 kinase 1323:1328 amino-terminal c-Jun kinase 1302:1328 amino-terminal c-Jun kinase (JNK) 1302:1334 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 10 3 dep CD14 1726:1729 arg1 both 1667:1670 both 1667:1670 both 1667:1670 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 2 4 link N-linked 315:322 arg1 carbohydrates 324:336 N-linked carbohydrates 315:336 N-linked carbohydrates of both MD-2 and TLR4 315:358 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 3 5 theme amino-terminal 465:478 arg1 ectodomain 480:489 the amino-terminal ectodomain 461:489 the amino-terminal ectodomain of human TLR4 461:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 5 theme amino-terminal 465:478 arg1 TLR4 500:503 human TLR4 494:503 human TLR4 494:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 6 6 theme mutants 1039:1045 arg1 ability 1002:1008 The ability 998:1008 The ability of the various glycosylation mutants to support cell activation 998:1072 The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. 11706042 6 7 theme various 1017:1023 arg1 mutants 1039:1045 the various glycosylation mutants 1013:1045 the various glycosylation mutants 1013:1045 The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. 11706042 4 8 theme MD-2 619:622 arg1 expression 605:614 cell surface expression 592:614 cell surface expression of MD-2 592:622 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 6 9 theme HeLa 1120:1123 arg1 cells 1125:1129 transiently transfected HeLa cells 1096:1129 transiently transfected HeLa cells 1096:1129 The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. 11706042 3 10 theme TLR4 500:503 arg1 ectodomain 480:489 the amino-terminal ectodomain 461:489 the amino-terminal ectodomain of human TLR4 461:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 10 theme TLR4 500:503 arg1 TLR4 500:503 human TLR4 494:503 human TLR4 494:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 7 11 theme IL-8 1272:1275 arg1 secretion 1277:1285 IL-8 secretion 1272:1285 IL-8 secretion 1272:1285 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 3 12 gly glycosylation 525:537 arg2 sites 539:543 9 N-linked glycosylation sites 514:543 9 N-linked glycosylation sites 514:543 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 12 gly glycosylation 525:537 arg2 9 514:514 9 514:514 9 514:514 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 6 13 theme transfected 1108:1118 arg1 cells 1125:1129 transiently transfected HeLa cells 1096:1129 transiently transfected HeLa cells 1096:1129 The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. 11706042 4 14 theme cell 592:595 arg1 expression 605:614 cell surface expression 592:614 cell surface expression of MD-2 592:622 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 9 15 theme Asn 1512:1514 arg1 mutants 1516:1522 the Asn mutants 1508:1522 the Asn mutants of MD-2 and TLR4 1508:1539 Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. 11706042 5 16 theme carbohydrates 942:954 arg1 role 920:923 a critical role 909:923 a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor 909:995 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 3 17 theme glycosylation 525:537 arg1 sites 539:543 9 N-linked glycosylation sites 514:543 9 N-linked glycosylation sites 514:543 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 7 18 theme luciferase 1242:1251 arg1 reporter 1253:1260 an interleukin-8 (IL-8) promoter-driven luciferase reporter 1202:1260 an interleukin-8 (IL-8) promoter-driven luciferase reporter 1202:1260 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 5 19 theme critical 911:918 arg1 role 920:923 a critical role 909:923 a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor 909:995 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 9 20 from reduction 1455:1463 arg1 activation 1468:1477 activation 1468:1477 activation resulting from expression of the Asn mutants of MD-2 and TLR4 1468:1539 Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. 11706042 4 21 theme cell 793:796 arg1 surface 798:804 the cell surface 789:804 the cell surface 789:804 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 8 22 theme Similar 1337:1343 arg1 results 1345:1351 Similar results 1337:1351 Similar results 1337:1351 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 5 23 theme LPS 984:986 arg1 receptor 988:995 the LPS receptor 980:995 the LPS receptor 980:995 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 1 24 theme multi-protein 141:153 arg1 receptor 127:134 The lipopolysaccharide (LPS) receptor 98:134 The lipopolysaccharide (LPS) receptor 98:134 The lipopolysaccharide (LPS) receptor is a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2. 11706042 1 24 theme multi-protein 141:153 arg1 complex 155:161 a multi-protein complex 139:161 a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2 139:224 The lipopolysaccharide (LPS) receptor is a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2. 11706042 10 25 theme functional 1827:1836 arg1 integrity 1838:1846 the functional integrity 1823:1846 the functional integrity of this receptor 1823:1863 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 0 26 theme lipopolysaccharide 69:86 arg1 receptor 88:95 a functional lipopolysaccharide receptor 56:95 a functional lipopolysaccharide receptor 56:95 MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor. 11706042 5 27 theme LPS 959:961 arg1 cross-linking 963:975 LPS cross-linking 959:975 LPS cross-linking to the LPS receptor 959:995 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 4 28 theme TLR4 701:704 arg1 mutants 706:712 TLR4 mutants 701:712 TLR4 mutants carrying substitutions in Asn(526) or Asn(575) 701:759 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 2 29 theme TLR4 355:358 arg1 carbohydrates 324:336 N-linked carbohydrates 315:336 N-linked carbohydrates of both MD-2 and TLR4 315:358 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 7 30 theme interleukin-8 1205:1217 arg1 reporter 1253:1260 an interleukin-8 (IL-8) promoter-driven luciferase reporter 1202:1260 an interleukin-8 (IL-8) promoter-driven luciferase reporter 1202:1260 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 4 31 theme site 674:677 arg1 presence 646:653 the presence 642:653 the presence of either N-linked site 642:677 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 2 32 theme MD-2 346:349 arg1 carbohydrates 324:336 N-linked carbohydrates 315:336 N-linked carbohydrates of both MD-2 and TLR4 315:358 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 0 33 theme MD-2 0:3 arg1 glycosylations 23:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor. 11706042 8 34 theme glycosylation 1416:1428 arg1 sites 1430:1434 three or more N-linked glycosylation sites 1393:1434 three or more N-linked glycosylation sites 1393:1434 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 0 35 theme TLR4 9:12 arg1 glycosylations 23:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor. 11706042 5 36 theme Re595 842:846 arg1 LPS 848:850 Re595 LPS 842:850 Re595 LPS (ASD-Re595 LPS) 842:866 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 5 36 theme Re595 842:846 arg1 LPS 863:865 ASD-Re595 LPS 853:865 ASD-Re595 LPS 853:865 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 7 37 theme LPS-induced 1176:1186 arg1 activation 1188:1197 LPS-induced activation 1176:1197 LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter 1176:1260 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 3 38 dep contains 386:393 arg1 whereas 453:459 whereas 453:459 whereas 453:459 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 39 link N-linked 516:523 arg1 sites 539:543 9 N-linked glycosylation sites 514:543 9 N-linked glycosylation sites 514:543 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 7 40 theme MD-2 1153:1156 arg1 mutant 1143:1148 The double mutant 1132:1148 The double mutant of MD-2 1132:1156 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 3 41 gly N-glycosylated 397:410 arg1 sites 412:416 2 N-glycosylated sites 395:416 2 N-glycosylated sites 395:416 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 6 42 theme glycosylation 1025:1037 arg1 mutants 1039:1045 the various glycosylation mutants 1013:1045 the various glycosylation mutants 1013:1045 The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. 11706042 9 43 theme mutants 1516:1522 arg1 expression 1494:1503 expression 1494:1503 expression of the Asn mutants of MD-2 and TLR4 1494:1539 Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. 11706042 4 44 theme Site-directed 546:558 arg1 studies 572:578 Site-directed mutagenesis studies 546:578 Site-directed mutagenesis studies 546:578 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 2 45 gly glycosylated 261:272 arg1 proteins 249:256 these proteins 243:256 these proteins 243:256 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 2 45 gly glycosylated 261:272 arg1 each 235:238 each 235:238 each 235:238 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 10 46 theme N-linked 1757:1764 arg1 TLR4 1789:1792 TLR4 1789:1792 TLR4 1789:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 46 theme N-linked 1757:1764 arg1 sites 1766:1770 N-linked sites 1757:1770 N-linked sites of both MD-2 and TLR4 1757:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 46 theme N-linked 1757:1764 arg1 MD-2 1780:1783 MD-2 1780:1783 MD-2 1780:1783 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 2 47 theme N-linked 315:322 arg1 carbohydrates 324:336 N-linked carbohydrates 315:336 N-linked carbohydrates of both MD-2 and TLR4 315:358 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 9 48 theme MD-2 1527:1530 arg1 mutants 1516:1522 the Asn mutants 1508:1522 the Asn mutants of MD-2 and TLR4 1508:1539 Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. 11706042 0 49 link N-linked 14:21 arg1 glycosylations 23:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor. 11706042 4 50 link N-linked 665:672 arg1 site 674:677 either N-linked site 658:677 either N-linked site 658:677 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 3 51 contain contains 505:512 arg2 sites 539:543 9 N-linked glycosylation sites 514:543 9 N-linked glycosylation sites 514:543 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 51 contain contains 505:512 arg1 ectodomain 480:489 the amino-terminal ectodomain 461:489 the amino-terminal ectodomain of human TLR4 461:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 51 contain contains 505:512 arg1 TLR4 500:503 human TLR4 494:503 human TLR4 494:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 9 52 theme TLR4 1536:1539 arg1 mutants 1516:1522 the Asn mutants 1508:1522 the Asn mutants of MD-2 and TLR4 1508:1539 Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. 11706042 10 53 theme functional 1618:1627 arg1 integrity 1629:1637 the functional integrity 1614:1637 the functional integrity of the LPS receptor 1614:1657 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 7 54 theme amino-terminal 1302:1315 arg1 JNK 1331:1333 JNK 1331:1333 JNK 1331:1333 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 7 54 theme amino-terminal 1302:1315 arg1 kinase 1323:1328 amino-terminal c-Jun kinase 1302:1328 amino-terminal c-Jun kinase (JNK) 1302:1334 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 4 55 theme surface 597:603 arg1 expression 605:614 cell surface expression 592:614 cell surface expression of MD-2 592:622 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 5 56 from role 920:923 arg1 cross-linking 963:975 LPS cross-linking 959:975 LPS cross-linking to the LPS receptor 959:995 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 3 57 theme human 494:498 arg1 TLR4 500:503 human TLR4 494:503 human TLR4 494:503 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 10 58 from TLR4 1742:1745 arg1 expression 1687:1696 the surface expression 1675:1696 the surface expression of at least three proteins 1675:1723 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 59 theme TLR4 1789:1792 arg1 TLR4 1789:1792 TLR4 1789:1792 TLR4 1789:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 59 theme TLR4 1789:1792 arg1 sites 1766:1770 N-linked sites 1757:1770 N-linked sites of both MD-2 and TLR4 1757:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 59 theme TLR4 1789:1792 arg1 MD-2 1780:1783 MD-2 1780:1783 MD-2 1780:1783 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 60 theme LPS 1646:1648 arg1 receptor 1650:1657 the LPS receptor 1642:1657 the LPS receptor 1642:1657 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 61 theme receptor 1650:1657 arg1 integrity 1629:1637 the functional integrity 1614:1637 the functional integrity of the LPS receptor 1614:1657 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 62 theme MD-2 1780:1783 arg1 TLR4 1789:1792 TLR4 1789:1792 TLR4 1789:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 62 theme MD-2 1780:1783 arg1 sites 1766:1770 N-linked sites 1757:1770 N-linked sites of both MD-2 and TLR4 1757:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 62 theme MD-2 1780:1783 arg1 MD-2 1780:1783 MD-2 1780:1783 MD-2 1780:1783 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 5 63 theme TLR4 937:940 arg1 carbohydrates 942:954 MD-2 and TLR4 carbohydrates 928:954 MD-2 and TLR4 carbohydrates 928:954 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 3 64 theme N-linked 516:523 arg1 sites 539:543 9 N-linked glycosylation sites 514:543 9 N-linked glycosylation sites 514:543 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 65 contain contains 386:393 arg1 MD-2 381:384 MD-2 381:384 MD-2 381:384 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 65 contain contains 386:393 arg2 sites 412:416 2 N-glycosylated sites 395:416 2 N-glycosylated sites 395:416 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 65 contain contains 386:393 arg2 MD-2 381:384 MD-2 381:384 MD-2 381:384 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 65 contain contains 386:393 arg1 positions 421:429 positions Asn(26) and Asn(114) 421:450 positions Asn(26) and Asn(114) 421:450 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 65 contain contains 386:393 arg1 Asn 443:445 Asn(114) 443:450 Asn(114) 443:450 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 65 contain contains 386:393 arg1 Asn 431:433 Asn(26) 431:437 Asn(26) 431:437 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 7 66 theme reporter 1253:1260 arg1 activation 1188:1197 LPS-induced activation 1176:1197 LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter 1176:1260 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 7 67 theme promoter-driven 1226:1240 arg1 reporter 1253:1260 an interleukin-8 (IL-8) promoter-driven luciferase reporter 1202:1260 an interleukin-8 (IL-8) promoter-driven luciferase reporter 1202:1260 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 0 68 theme functional 58:67 arg1 receptor 88:95 a functional lipopolysaccharide receptor 56:95 a functional lipopolysaccharide receptor 56:95 MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor. 11706042 5 69 theme MD-2 928:931 arg1 carbohydrates 942:954 MD-2 and TLR4 carbohydrates 928:954 MD-2 and TLR4 carbohydrates 928:954 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 4 70 from substitutions 723:735 arg1 575 756:758 575 756:758 575 756:758 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 4 70 from substitutions 723:735 arg1 526 744:746 526 744:746 526 744:746 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 4 70 from substitutions 723:735 arg1 Asn 752:754 Asn 752:754 Asn(575) 752:759 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 4 70 from substitutions 723:735 arg1 Asn 740:742 Asn 740:742 Asn(526) 740:747 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 8 71 theme TLR4 1372:1375 arg1 mutants 1377:1383 TLR4 mutants 1372:1383 TLR4 mutants lacking three or more N-linked glycosylation sites 1372:1434 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 6 72 theme cell 1058:1061 arg1 activation 1063:1072 cell activation 1058:1072 cell activation 1058:1072 The ability of the various glycosylation mutants to support cell activation was also evaluated in transiently transfected HeLa cells. 11706042 4 73 contain carrying 714:721 arg1 mutants 706:712 TLR4 mutants 701:712 TLR4 mutants carrying substitutions in Asn(526) or Asn(575) 701:759 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 4 73 contain carrying 714:721 arg2 substitutions 723:735 substitutions 723:735 substitutions in Asn(526) or Asn(575) 723:759 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 8 74 gly glycosylation 1416:1428 arg2 more 1402:1405 more 1402:1405 more 1402:1405 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 8 74 gly glycosylation 1416:1428 arg2 three 1393:1397 three 1393:1397 three 1393:1397 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 8 74 gly glycosylation 1416:1428 arg2 sites 1430:1434 three or more N-linked glycosylation sites 1393:1434 three or more N-linked glycosylation sites 1393:1434 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 5 75 theme LPS 848:850 arg1 derivative 828:837 a UV-activated derivative 813:837 a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) 813:866 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 10 76 theme receptor 1856:1863 arg1 integrity 1838:1846 the functional integrity 1823:1846 the functional integrity of this receptor 1823:1863 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 8 77 theme N-linked 1407:1414 arg1 sites 1430:1434 three or more N-linked glycosylation sites 1393:1434 three or more N-linked glycosylation sites 1393:1434 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 3 78 theme N-glycosylated 397:410 arg1 sites 412:416 2 N-glycosylated sites 395:416 2 N-glycosylated sites 395:416 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 5 79 theme UV-activated 815:826 arg1 derivative 828:837 a UV-activated derivative 813:837 a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) 813:866 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 4 80 theme N-linked 665:672 arg1 site 674:677 either N-linked site 658:677 either N-linked site 658:677 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 0 81 theme N-linked 14:21 arg1 glycosylations 23:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations 0:36 MD-2 and TLR4 N-linked glycosylations are important for a functional lipopolysaccharide receptor. 11706042 7 82 dep support 1168:1174 arg1 activate 1293:1300 activate 1293:1300 to activate amino-terminal c-Jun kinase (JNK) 1290:1334 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 7 82 dep support 1168:1174 arg1 induce 1265:1270 induce 1265:1270 to induce IL-8 secretion 1262:1285 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 1 83 theme lipopolysaccharide 102:119 arg1 receptor 127:134 The lipopolysaccharide (LPS) receptor 98:134 The lipopolysaccharide (LPS) receptor 98:134 The lipopolysaccharide (LPS) receptor is a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2. 11706042 1 83 theme lipopolysaccharide 102:119 arg1 complex 155:161 a multi-protein complex 139:161 a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2 139:224 The lipopolysaccharide (LPS) receptor is a multi-protein complex that consists of at least three proteins, CD14, TLR4, and MD-2. 11706042 10 84 theme surface 1679:1685 arg1 expression 1687:1696 the surface expression 1675:1696 the surface expression of at least three proteins 1675:1723 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 3 85 dep positions 421:429 arg1 positions 421:429 positions Asn(26) and Asn(114) 421:450 positions Asn(26) and Asn(114) 421:450 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 85 dep positions 421:429 arg1 Asn 443:445 Asn(114) 443:450 Asn(114) 443:450 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 3 85 dep positions 421:429 arg1 Asn 431:433 Asn(26) 431:437 Asn(26) 431:437 We demonstrate that MD-2 contains 2 N-glycosylated sites at positions Asn(26) and Asn(114), whereas the amino-terminal ectodomain of human TLR4 contains 9 N-linked glycosylation sites. 11706042 10 86 theme proteins 1716:1723 arg1 expression 1687:1696 the surface expression 1675:1696 the surface expression of at least three proteins 1675:1723 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 9 87 with co-expression 1570:1582 arg1 CD14 1589:1592 CD14 1589:1592 CD14 1589:1592 Surprisingly, the reduction in activation resulting from expression of the Asn mutants of MD-2 and TLR4 can be partially reversed by co-expression with CD14. 11706042 2 88 theme functional 296:305 arg1 role 307:310 the functional role 292:310 the functional role of N-linked carbohydrates of both MD-2 and TLR4 292:358 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 7 89 theme double 1136:1141 arg1 mutant 1143:1148 The double mutant 1132:1148 The double mutant of MD-2 1132:1156 The double mutant of MD-2 failed to support LPS-induced activation of an interleukin-8 (IL-8) promoter-driven luciferase reporter to induce IL-8 secretion or to activate amino-terminal c-Jun kinase (JNK). 11706042 2 90 theme carbohydrates 324:336 arg1 role 307:310 the functional role 292:310 the functional role of N-linked carbohydrates of both MD-2 and TLR4 292:358 Because each of these proteins is glycosylated, we have examined the functional role of N-linked carbohydrates of both MD-2 and TLR4. 11706042 5 91 theme cross-linking 871:883 arg1 assays 885:890 cross-linking assays 871:890 cross-linking assays 871:890 Using a UV-activated derivative of Re595 LPS (ASD-Re595 LPS) in cross-linking assays, we demonstrated a critical role of MD-2 and TLR4 carbohydrates in LPS cross-linking to the LPS receptor. 11706042 8 92 link N-linked 1407:1414 arg1 sites 1430:1434 three or more N-linked glycosylation sites 1393:1434 three or more N-linked glycosylation sites 1393:1434 Similar results were observed with TLR4 mutants lacking three or more N-linked glycosylation sites. 11706042 4 93 theme mutagenesis 560:570 arg1 studies 572:578 Site-directed mutagenesis studies 546:578 Site-directed mutagenesis studies 546:578 Site-directed mutagenesis studies showed that cell surface expression of MD-2 did not depend on the presence of either N-linked site, whereas in contrast, TLR4 mutants carrying substitutions in Asn(526) or Asn(575) failed to be transported to the cell surface. 11706042 10 94 link N-linked 1757:1764 arg1 TLR4 1789:1792 TLR4 1789:1792 TLR4 1789:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 94 link N-linked 1757:1764 arg1 sites 1766:1770 N-linked sites 1757:1770 N-linked sites of both MD-2 and TLR4 1757:1792 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 11706042 10 94 link N-linked 1757:1764 arg1 MD-2 1780:1783 MD-2 1780:1783 MD-2 1780:1783 This suggests that the functional integrity of the LPS receptor depends both on the surface expression of at least three proteins, CD14, MD-2, and TLR4, and that N-linked sites of both MD-2 and TLR4 are essential in maintaining the functional integrity of this receptor. 10639192 1 0 theme human 128:132 arg1 gland 143:147 the human prostate gland 124:147 the human prostate gland 124:147 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 7 1 theme human 991:995 arg1 enzyme 997:1002 the human enzyme 987:1002 the human enzyme 987:1002 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 2 2 theme crystal 310:316 arg1 structure 318:326 its crystal structure 306:326 its crystal structure 306:326 METHODS: Human PAP, purified from semen, was crystallized using polyethylene glycol as the precipitant and its crystal structure was determined using X-ray diffraction. 10639192 6 3 theme complex 750:756 arg1 chains 758:763 complex chains 750:763 complex chains 750:763 It appears that high-mannose chains are attached to Asn 62 and Asp 301, while complex chains are at Asn 188. 10639192 6 4 attach attached 712:719 arg1 Asn 724:726 Asn 62 724:729 Asn 62 724:729 It appears that high-mannose chains are attached to Asn 62 and Asp 301, while complex chains are at Asn 188. 10639192 6 4 attach attached 712:719 arg1 Asp 735:737 Asp 301 735:741 Asp 301 735:741 It appears that high-mannose chains are attached to Asn 62 and Asp 301, while complex chains are at Asn 188. 10639192 6 4 attach attached 712:719 arg2 chains 701:706 high-mannose chains 688:706 high-mannose chains 688:706 It appears that high-mannose chains are attached to Asn 62 and Asp 301, while complex chains are at Asn 188. 10639192 9 5 theme protein 1185:1191 arg1 carbohydrates 1164:1176 The carbohydrates 1160:1176 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system 1160:1263 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 7 6 theme rat 848:850 arg1 PAP 852:854 rat PAP 848:854 rat PAP 848:854 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 5 7 gly glycosylation 651:663 arg2 sites 665:669 the possible glycosylation sites 638:669 the possible glycosylation sites 638:669 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 9 8 theme complex 1297:1303 arg1 carbohydrates 1305:1317 complex carbohydrates attachment 1297:1328 complex carbohydrates attachment 1297:1328 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 7 9 theme three-dimensional 816:832 arg1 structures 834:843 the three-dimensional structures 812:843 the three-dimensional structures of rat PAP and human PAP 812:868 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 5 10 link N-linked 592:599 arg1 carbohydrates 601:613 N-linked carbohydrates 592:613 N-linked carbohydrates 592:613 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 9 11 theme expression 1247:1256 arg1 system 1258:1263 the baculovirus expression system 1231:1263 the baculovirus expression system 1231:1263 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 9 12 theme carbohydrates 1305:1317 arg1 site 1289:1292 the site 1285:1292 the site of complex carbohydrates attachment 1285:1328 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 9 12 theme carbohydrates 1305:1317 arg1 carbohydrates 1305:1317 complex carbohydrates attachment 1297:1328 complex carbohydrates attachment 1297:1328 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 7 13 theme structures 834:843 arg1 similarity 798:807 The similarity 794:807 The similarity of the three-dimensional structures of rat PAP and human PAP 794:868 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 1 14 theme Prostatic 68:76 arg1 BACKGROUND 56:65 BACKGROUND 56:65 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 56:197 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 14 theme Prostatic 68:76 arg1 product 113:119 a major product 105:119 a major product of the human prostate gland 105:147 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 14 theme Prostatic 68:76 arg1 hPAP 96:99 hPAP 96:99 hPAP 96:99 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 14 theme Prostatic 68:76 arg1 phosphatase 83:93 Prostatic acid phosphatase 68:93 Prostatic acid phosphatase (hPAP) 68:100 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 4 15 theme catalytic 558:566 arg1 residues 568:575 its catalytic residues 554:575 its catalytic residues 554:575 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 10639192 1 16 theme gland 143:147 arg1 BACKGROUND 56:65 BACKGROUND 56:65 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 56:197 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 16 theme gland 143:147 arg1 product 113:119 a major product 105:119 a major product of the human prostate gland 105:147 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 16 theme gland 143:147 arg1 phosphatase 83:93 Prostatic acid phosphatase 68:93 Prostatic acid phosphatase (hPAP) 68:100 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 8 17 theme N-linked 1099:1106 arg1 chains 1121:1126 N-linked carbohydrate chains 1099:1126 N-linked carbohydrate chains 1099:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 6 18 theme high-mannose 688:699 arg1 chains 701:706 high-mannose chains 688:706 high-mannose chains 688:706 It appears that high-mannose chains are attached to Asn 62 and Asp 301, while complex chains are at Asn 188. 10639192 1 19 theme acid 78:81 arg1 BACKGROUND 56:65 BACKGROUND 56:65 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 56:197 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 19 theme acid 78:81 arg1 product 113:119 a major product 105:119 a major product of the human prostate gland 105:147 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 19 theme acid 78:81 arg1 hPAP 96:99 hPAP 96:99 hPAP 96:99 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 19 theme acid 78:81 arg1 phosphatase 83:93 Prostatic acid phosphatase 68:93 Prostatic acid phosphatase (hPAP) 68:100 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 5 20 attach present 619:625 arg2 carbohydrates 601:613 N-linked carbohydrates 592:613 N-linked carbohydrates 592:613 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 5 20 attach present 619:625 arg1 sites 665:669 the possible glycosylation sites 638:669 the possible glycosylation sites 638:669 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 5 20 attach present 619:625 arg1 each 630:633 each 630:633 each 630:633 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 8 21 theme chains 1121:1126 arg1 chains 1121:1126 N-linked carbohydrate chains 1099:1126 N-linked carbohydrate chains 1099:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 8 21 theme chains 1121:1126 arg1 sites 1090:1094 attachment sites 1079:1094 attachment sites of N-linked carbohydrate chains 1079:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 9 22 theme prostate 1209:1216 arg1 cells 1218:1222 the prostate cells 1205:1222 the prostate cells 1205:1222 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 7 23 theme rat 954:956 arg1 enzyme 958:963 the rat enzyme 950:963 the rat enzyme 950:963 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 8 24 link N-linked 1099:1106 arg1 chains 1121:1126 N-linked carbohydrate chains 1099:1126 N-linked carbohydrate chains 1099:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 2 25 theme X-ray 349:353 arg1 diffraction 355:365 X-ray diffraction 349:365 X-ray diffraction 349:365 METHODS: Human PAP, purified from semen, was crystallized using polyethylene glycol as the precipitant and its crystal structure was determined using X-ray diffraction. 10639192 0 26 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human prostatic acid phosphatase 0:52 Crystal structure of human prostatic acid phosphatase . 10639192 0 27 theme human 21:25 arg1 phosphatase 42:52 human prostatic acid phosphatase 21:52 human prostatic acid phosphatase 21:52 Crystal structure of human prostatic acid phosphatase . 10639192 5 28 theme N-linked 592:599 arg1 carbohydrates 601:613 N-linked carbohydrates 592:613 N-linked carbohydrates 592:613 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 3 29 dep = 437:437 arg1 % 441:441 27% 439:441 27% 439:441 The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. 10639192 3 30 theme = 419:419 arg1 R 417:417 R 417:417 R = 16% 417:423 The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. 10639192 7 31 theme PAP 866:868 arg1 structures 834:843 the three-dimensional structures 812:843 the three-dimensional structures of rat PAP and human PAP 812:868 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 4 32 theme hPAP 470:473 arg1 structure 457:465 The structure 453:465 The structure of hPAP 453:473 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 10639192 4 32 theme hPAP 470:473 arg1 similar 478:484 similar 478:484 similar 478:484 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 10639192 3 33 dep R 429:429 arg1 free 431:434 free 431:434 free 431:434 The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. 10639192 5 34 from each 630:633 arg1 present 619:625 present 619:625 present 619:625 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 1 35 theme prostate 134:141 arg1 gland 143:147 the human prostate gland 124:147 the human prostate gland 124:147 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 8 36 theme carbohydrate 1141:1152 arg1 type 1154:1157 a given carbohydrate type 1133:1157 a given carbohydrate type 1133:1157 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 0 37 theme acid 37:40 arg1 phosphatase 42:52 human prostatic acid phosphatase 21:52 human prostatic acid phosphatase 21:52 Crystal structure of human prostatic acid phosphatase . 10639192 7 38 theme mechanism 927:935 arg1 analyses 901:908 the mechanistic analyses 885:908 the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme 885:963 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 2 39 theme Human 208:212 arg1 PAP 214:216 Human PAP 208:216 Human PAP 208:216 METHODS: Human PAP, purified from semen, was crystallized using polyethylene glycol as the precipitant and its crystal structure was determined using X-ray diffraction. 10639192 0 40 theme prostatic 27:35 arg1 phosphatase 42:52 human prostatic acid phosphatase 21:52 human prostatic acid phosphatase 21:52 Crystal structure of human prostatic acid phosphatase . 10639192 9 41 theme baculovirus 1235:1245 arg1 system 1258:1263 the baculovirus expression system 1231:1263 the baculovirus expression system 1231:1263 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 8 42 theme sites 1090:1094 arg1 correlation 1064:1074 the correlation 1060:1074 the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type 1060:1157 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 8 43 theme attachment 1079:1088 arg1 chains 1121:1126 N-linked carbohydrate chains 1099:1126 N-linked carbohydrate chains 1099:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 8 43 theme attachment 1079:1088 arg1 sites 1090:1094 attachment sites 1079:1094 attachment sites of N-linked carbohydrate chains 1079:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 8 44 theme carbohydrate 1108:1119 arg1 chains 1121:1126 N-linked carbohydrate chains 1099:1126 N-linked carbohydrate chains 1099:1126 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 8 45 theme crystallographic 1030:1045 arg1 data 1047:1050 The crystallographic data 1026:1050 The crystallographic data 1026:1050 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 3 46 theme A 401:401 arg1 resolution 403:412 3.1 A resolution 397:412 3.1 A resolution 397:412 The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. 10639192 7 47 theme mechanistic 889:899 arg1 analyses 901:908 the mechanistic analyses 885:908 the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme 885:963 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 0 48 theme phosphatase 42:52 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human prostatic acid phosphatase 0:52 Crystal structure of human prostatic acid phosphatase . 10639192 7 49 theme catalytic 917:925 arg1 mechanism 927:935 the catalytic mechanism 913:935 the catalytic mechanism proposed for the rat enzyme 913:963 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 8 50 with correlation 1064:1074 arg1 type 1154:1157 a given carbohydrate type 1133:1157 a given carbohydrate type 1133:1157 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 9 51 dep carbohydrates 1305:1317 arg1 attachment 1319:1328 attachment 1319:1328 complex carbohydrates attachment 1297:1328 The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment. 10639192 8 52 theme given 1135:1139 arg1 type 1154:1157 a given carbohydrate type 1133:1157 a given carbohydrate type 1133:1157 The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. 10639192 1 53 theme major 107:111 arg1 BACKGROUND 56:65 BACKGROUND 56:65 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 56:197 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 53 theme major 107:111 arg1 product 113:119 a major product 105:119 a major product of the human prostate gland 105:147 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 1 53 theme major 107:111 arg1 phosphatase 83:93 Prostatic acid phosphatase 68:93 Prostatic acid phosphatase (hPAP) 68:100 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 2 54 theme polyethylene 263:274 arg1 glycol 276:281 polyethylene glycol 263:281 polyethylene glycol 263:281 METHODS: Human PAP, purified from semen, was crystallized using polyethylene glycol as the precipitant and its crystal structure was determined using X-ray diffraction. 10639192 2 54 theme polyethylene 263:274 arg1 precipitant 290:300 the precipitant 286:300 the precipitant 286:300 METHODS: Human PAP, purified from semen, was crystallized using polyethylene glycol as the precipitant and its crystal structure was determined using X-ray diffraction. 10639192 4 55 theme other 497:501 arg1 phosphatases 519:530 other known histidine phosphatases 497:530 other known histidine phosphatases 497:530 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 10639192 7 56 theme human 860:864 arg1 PAP 866:868 human PAP 860:868 human PAP 860:868 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 5 57 located present 619:625 arg2 carbohydrates 601:613 N-linked carbohydrates 592:613 N-linked carbohydrates 592:613 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 5 57 located present 619:625 arg1 sites 665:669 the possible glycosylation sites 638:669 the possible glycosylation sites 638:669 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 5 57 located present 619:625 arg1 each 630:633 each 630:633 each 630:633 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 5 58 theme possible 642:649 arg1 sites 665:669 the possible glycosylation sites 638:669 the possible glycosylation sites 638:669 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 3 59 dep = 419:419 arg1 % 423:423 16% 421:423 16% 421:423 The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. 10639192 1 60 theme physiological 158:170 arg1 substrate 172:180 its physiological substrate 154:180 its physiological substrate 154:180 BACKGROUND: Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. 10639192 3 61 theme = 437:437 arg1 R 429:429 R 429:429 R(free) = 27% 429:441 The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. 10639192 4 62 theme histidine 509:517 arg1 phosphatases 519:530 other known histidine phosphatases 497:530 other known histidine phosphatases 497:530 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 10639192 5 63 theme glycosylation 651:663 arg1 sites 665:669 the possible glycosylation sites 638:669 the possible glycosylation sites 638:669 N-linked carbohydrates are present at each of the possible glycosylation sites. 10639192 7 64 theme PAP 852:854 arg1 structures 834:843 the three-dimensional structures 812:843 the three-dimensional structures of rat PAP and human PAP 812:868 CONCLUSIONS: The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. 10639192 4 65 theme residues 568:575 arg1 positions 541:549 the positions 537:549 the positions of its catalytic residues 537:575 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 10639192 4 66 theme known 503:507 arg1 phosphatases 519:530 other known histidine phosphatases 497:530 other known histidine phosphatases 497:530 RESULTS: The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. 11502179 10 0 theme soluble 1500:1506 arg1 ectodomain 1508:1517 the soluble ectodomain 1496:1517 the soluble ectodomain of the TSH receptor purified in a functionally competent conformation 1496:1587 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 10 0 theme soluble 1500:1506 arg1 receptor 1530:1537 the TSH receptor 1522:1537 the TSH receptor purified in a functionally competent conformation 1522:1587 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 1 1 theme TSH 171:173 arg1 receptor 175:182 the human TSH receptor 161:182 the human TSH receptor 161:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 9 2 theme immunoglobulins 1382:1396 arg1 amounts 1365:1371 the amounts 1361:1371 the amounts of these immunoglobulins 1361:1396 It allowed computation of the amounts of these immunoglobulins in patient's serum, giving values up to 10 microg/mL. 11502179 9 2 theme immunoglobulins 1382:1396 arg1 immunoglobulins 1382:1396 these immunoglobulins 1376:1396 these immunoglobulins 1376:1396 It allowed computation of the amounts of these immunoglobulins in patient's serum, giving values up to 10 microg/mL. 11502179 7 3 theme site 1064:1067 arg1 A 1069:1069 site A 1064:1069 site A 1064:1069 The significance of site A, whose affinity is much higher than for the holoreceptor at the surface of intact cells, remains to be clarified. 11502179 1 4 theme receptor 175:182 arg1 ectodomain 147:156 The amino-terminal ectodomain 128:156 The amino-terminal ectodomain of the human TSH receptor 128:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 4 theme receptor 175:182 arg1 molecule 274:281 a glycosylphosphatidylinositol-anchored molecule 234:281 a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus 234:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 4 theme receptor 175:182 arg1 receptor 175:182 the human TSH receptor 161:182 the human TSH receptor 161:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 6 5 theme site 1022:1025 arg1 B 1027:1027 site B 1022:1027 site B 1022:1027 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 0 6 theme thyrotropin 106:116 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 2 7 attach released 378:385 arg1 cells 396:400 the cells 392:400 the cells 392:400 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 2 7 attach released 378:385 arg2 ectodomain 358:367 The soluble ectodomain 346:367 The soluble ectodomain 346:367 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 10 8 theme structural 1680:1689 arg1 studies 1691:1697 structural studies 1680:1697 structural studies 1680:1697 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 8 9 from activity 1271:1278 arg1 patients 1304:1311 patients 1304:1311 patients with Graves' disease 1304:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 0 10 theme human 100:104 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 6 11 theme binding 968:974 arg1 sites 976:980 high-affinity binding sites 954:980 high-affinity binding sites 954:980 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 1 12 theme 10-residue 296:305 arg1 tag 317:319 a 10-residue histidine tag 294:319 a 10-residue histidine tag close to its C terminus 294:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 5 13 theme specific 814:821 arg1 binding 823:829 specific binding 814:829 specific binding of (125)I-labeled TSH 814:851 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 6 14 theme high-affinity 954:966 arg1 sites 976:980 high-affinity binding sites 954:980 high-affinity binding sites 954:980 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 8 15 theme thyroid 1251:1257 arg1 activity 1271:1278 the thyroid stimulating activity 1247:1278 the thyroid stimulating activity of immunoglobulins from patients with Graves' disease 1247:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 6 16 theme d 1032:1032 arg1 nM 1040:1041 K(d) 0.83 nM 1030:1041 K(d) 0.83 nM 1030:1041 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 10 17 theme interactions 1618:1629 arg1 studies 1603:1609 direct studies 1596:1609 direct studies of its interactions with TSH and autoantibodies 1596:1657 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 5 18 theme binding 918:924 arg1 experiments 926:936 classical saturation binding experiments 897:936 classical saturation binding experiments 897:936 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 0 19 theme receptor 118:125 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 0 19 theme receptor 118:125 arg1 domain 86:91 a soluble bioactive amino-terminal extracellular domain 37:91 a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor 37:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 6 20 dep identified 987:996 arg1 A 1004:1004 site A 999:1004 site A 999:1004 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 5 21 theme I-labeled 839:847 arg1 TSH 849:851 (125)I-labeled TSH 834:851 (125)I-labeled TSH 834:851 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 7 22 theme intact 1146:1151 arg1 cells 1153:1157 intact cells 1146:1157 intact cells 1146:1157 The significance of site A, whose affinity is much higher than for the holoreceptor at the surface of intact cells, remains to be clarified. 11502179 4 23 theme sites 715:719 arg1 four 669:672 four 669:672 four 669:672 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 4 23 theme sites 715:719 arg1 sites 715:719 the six potential N-glycosylation sites 681:719 the six potential N-glycosylation sites 681:719 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 10 24 theme competent 1566:1574 arg1 conformation 1576:1587 a functionally competent conformation 1551:1587 a functionally competent conformation 1551:1587 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 7 25 theme A 1069:1069 arg1 significance 1048:1059 The significance 1044:1059 The significance 1044:1059 The significance of site A, whose affinity is much higher than for the holoreceptor at the surface of intact cells, remains to be clarified. 11502179 8 26 from patients 1304:1311 arg1 activity 1271:1278 the thyroid stimulating activity 1247:1278 the thyroid stimulating activity of immunoglobulins from patients with Graves' disease 1247:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 8 26 from patients 1304:1311 arg1 immunoglobulins 1283:1297 immunoglobulins 1283:1297 immunoglobulins from patients with Graves' disease 1283:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 5 27 theme purified 776:783 arg1 ectodomain 793:802 the purified soluble ectodomain 772:802 the purified soluble ectodomain 772:802 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 2 28 theme apparent 483:490 arg1 homogeneity 492:502 apparent homogeneity 483:502 apparent homogeneity 483:502 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 1 29 theme histidine 307:315 arg1 tag 317:319 a 10-residue histidine tag 294:319 a 10-residue histidine tag close to its C terminus 294:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 4 30 gly N-glycosylation 699:713 arg2 sites 715:719 the six potential N-glycosylation sites 681:719 the six potential N-glycosylation sites 681:719 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 4 30 gly N-glycosylation 699:713 arg2 six 685:687 six 685:687 six 685:687 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 5 31 theme ectodomain 793:802 arg1 proportion 758:767 A proportion 756:767 A proportion of the purified soluble ectodomain 756:802 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 5 32 theme first 871:875 arg1 performance 882:892 the first time performance 867:892 the first time performance of classical saturation binding experiments 867:936 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 5 33 theme classical 897:905 arg1 experiments 926:936 classical saturation binding experiments 897:936 classical saturation binding experiments 897:936 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 6 34 theme sites 976:980 arg1 classes 943:949 Two classes 939:949 Two classes of high-affinity binding sites 939:980 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 5 35 theme time 877:880 arg1 performance 882:892 the first time performance 867:892 the first time performance of classical saturation binding experiments 867:936 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 6 36 theme d 1009:1009 arg1 nM 1018:1019 K(d) 0.014 nM 1007:1019 K(d) 0.014 nM 1007:1019 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 4 37 theme four 669:672 arg1 identification 651:664 the identification 647:664 the identification of four out of the six potential N-glycosylation sites 647:719 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 6 38 dep A 1004:1004 arg1 nM 1040:1041 K(d) 0.83 nM 1030:1041 K(d) 0.83 nM 1030:1041 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 6 38 dep A 1004:1004 arg1 nM 1018:1019 K(d) 0.014 nM 1007:1019 K(d) 0.014 nM 1007:1019 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 6 38 dep A 1004:1004 arg1 B 1027:1027 site B 1022:1027 site B 1022:1027 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 3 39 theme mass 615:618 arg1 spectrometry 620:631 mass spectrometry 615:631 mass spectrometry 615:631 Two nanomoles of material was obtained, which was suitable for analysis by mass spectrometry. 11502179 0 40 theme soluble 39:45 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 0 40 theme soluble 39:45 arg1 domain 86:91 a soluble bioactive amino-terminal extracellular domain 37:91 a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor 37:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 3 41 theme material 557:564 arg1 nanomoles 544:552 Two nanomoles 540:552 Two nanomoles of material 540:564 Two nanomoles of material was obtained, which was suitable for analysis by mass spectrometry. 11502179 2 42 with treatment 405:413 arg1 C 465:465 a glycosylphosphatidylinositol-phospholipase C 420:465 a glycosylphosphatidylinositol-phospholipase C 420:465 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 4 43 theme N-glycosylation 699:713 arg1 sites 715:719 the six potential N-glycosylation sites 681:719 the six potential N-glycosylation sites 681:719 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 2 44 theme glycosylphosphatidylinositol-phospholipase 422:463 arg1 C 465:465 a glycosylphosphatidylinositol-phospholipase C 420:465 a glycosylphosphatidylinositol-phospholipase C 420:465 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 5 45 theme TSH 849:851 arg1 binding 823:829 specific binding 814:829 specific binding of (125)I-labeled TSH 814:851 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 6 46 theme K 1007:1007 arg1 nM 1018:1019 K(d) 0.014 nM 1007:1019 K(d) 0.014 nM 1007:1019 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 5 47 theme soluble 785:791 arg1 ectodomain 793:802 the purified soluble ectodomain 772:802 the purified soluble ectodomain 772:802 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 5 48 theme saturation 907:916 arg1 experiments 926:936 classical saturation binding experiments 897:936 classical saturation binding experiments 897:936 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 1 49 theme CHO 221:223 arg1 cells 225:229 CHO cells 221:229 CHO cells 221:229 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 2 50 theme cobalt-Sepharose 507:522 arg1 chromatography 524:537 cobalt-Sepharose chromatography 507:537 cobalt-Sepharose chromatography 507:537 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 1 51 theme C 334:334 arg1 terminus 336:343 its C terminus 330:343 its C terminus 330:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 0 52 theme extracellular 72:84 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 0 52 theme extracellular 72:84 arg1 domain 86:91 a soluble bioactive amino-terminal extracellular domain 37:91 a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor 37:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 8 53 theme stimulating 1259:1269 arg1 activity 1271:1278 the thyroid stimulating activity 1247:1278 the thyroid stimulating activity of immunoglobulins from patients with Graves' disease 1247:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 0 54 theme bioactive 47:55 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 0 54 theme bioactive 47:55 arg1 domain 86:91 a soluble bioactive amino-terminal extracellular domain 37:91 a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor 37:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 10 55 with interactions 1618:1629 arg1 autoantibodies 1644:1657 autoantibodies 1644:1657 autoantibodies 1644:1657 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 10 55 with interactions 1618:1629 arg1 TSH 1636:1638 TSH 1636:1638 TSH 1636:1638 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 1 56 theme amino-terminal 132:145 arg1 ectodomain 147:156 The amino-terminal ectodomain 128:156 The amino-terminal ectodomain of the human TSH receptor 128:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 56 theme amino-terminal 132:145 arg1 molecule 274:281 a glycosylphosphatidylinositol-anchored molecule 234:281 a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus 234:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 56 theme amino-terminal 132:145 arg1 receptor 175:182 the human TSH receptor 161:182 the human TSH receptor 161:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 6 57 theme site 999:1002 arg1 A 1004:1004 site A 999:1004 site A 999:1004 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 10 58 theme available 1478:1486 arg1 assays 1488:1493 all currently available assays 1464:1493 all currently available assays 1464:1493 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 8 59 with patients 1304:1311 arg1 disease 1326:1332 Graves' disease 1318:1332 Graves' disease 1318:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 10 60 theme receptor 1530:1537 arg1 ectodomain 1508:1517 the soluble ectodomain 1496:1517 the soluble ectodomain of the TSH receptor purified in a functionally competent conformation 1496:1587 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 10 60 theme receptor 1530:1537 arg1 receptor 1530:1537 the TSH receptor 1522:1537 the TSH receptor purified in a functionally competent conformation 1522:1587 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 2 61 theme soluble 350:356 arg1 ectodomain 358:367 The soluble ectodomain 346:367 The soluble ectodomain 346:367 The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. 11502179 1 62 theme cells 225:229 arg1 surface 210:216 the surface 206:216 the surface of CHO cells 206:229 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 8 63 theme purified 1189:1196 arg1 capable 1213:1219 capable 1213:1219 capable 1213:1219 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 8 63 theme purified 1189:1196 arg1 ectodomain 1198:1207 The purified ectodomain 1185:1207 The purified ectodomain 1185:1207 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 8 64 theme immunoglobulins 1283:1297 arg1 activity 1271:1278 the thyroid stimulating activity 1247:1278 the thyroid stimulating activity of immunoglobulins from patients with Graves' disease 1247:1332 The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. 11502179 9 65 theme amounts 1365:1371 arg1 computation 1346:1356 computation 1346:1356 computation of the amounts of these immunoglobulins 1346:1396 It allowed computation of the amounts of these immunoglobulins in patient's serum, giving values up to 10 microg/mL. 11502179 0 66 theme domain 86:91 arg1 characterization 17:32 characterization 17:32 characterization 17:32 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 0 66 theme domain 86:91 arg1 Purification 0:11 Purification 0:11 Purification 0:11 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 4 67 theme potential 689:697 arg1 sites 715:719 the six potential N-glycosylation sites 681:719 the six potential N-glycosylation sites 681:719 This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. 11502179 1 68 contain containing 283:292 arg1 ectodomain 147:156 The amino-terminal ectodomain 128:156 The amino-terminal ectodomain of the human TSH receptor 128:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 68 contain containing 283:292 arg2 tag 317:319 a 10-residue histidine tag 294:319 a 10-residue histidine tag close to its C terminus 294:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 68 contain containing 283:292 arg1 molecule 274:281 a glycosylphosphatidylinositol-anchored molecule 234:281 a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus 234:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 68 contain containing 283:292 arg1 receptor 175:182 the human TSH receptor 161:182 the human TSH receptor 161:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 10 69 theme TSH 1526:1528 arg1 receptor 1530:1537 the TSH receptor 1522:1537 the TSH receptor purified in a functionally competent conformation 1522:1587 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 5 70 theme experiments 926:936 arg1 performance 882:892 the first time performance 867:892 the first time performance of classical saturation binding experiments 867:936 A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. 11502179 6 71 theme K 1030:1030 arg1 nM 1040:1041 K(d) 0.83 nM 1030:1041 K(d) 0.83 nM 1030:1041 Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. 11502179 0 72 theme amino-terminal 57:70 arg1 receptor 118:125 the human thyrotropin receptor 96:125 the human thyrotropin receptor 96:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 0 72 theme amino-terminal 57:70 arg1 domain 86:91 a soluble bioactive amino-terminal extracellular domain 37:91 a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor 37:125 Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. 11502179 7 73 theme cells 1153:1157 arg1 surface 1135:1141 the surface 1131:1141 the surface of intact cells 1131:1157 The significance of site A, whose affinity is much higher than for the holoreceptor at the surface of intact cells, remains to be clarified. 11502179 1 74 theme human 165:169 arg1 receptor 175:182 the human TSH receptor 161:182 the human TSH receptor 161:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 10 75 theme direct 1596:1601 arg1 studies 1603:1609 direct studies 1596:1609 direct studies of its interactions with TSH and autoantibodies 1596:1657 Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. 11502179 1 76 theme glycosylphosphatidylinositol-anchored 236:272 arg1 ectodomain 147:156 The amino-terminal ectodomain 128:156 The amino-terminal ectodomain of the human TSH receptor 128:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 76 theme glycosylphosphatidylinositol-anchored 236:272 arg1 molecule 274:281 a glycosylphosphatidylinositol-anchored molecule 234:281 a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus 234:343 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 11502179 1 76 theme glycosylphosphatidylinositol-anchored 236:272 arg1 receptor 175:182 the human TSH receptor 161:182 the human TSH receptor 161:182 The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. 3463996 7 0 theme active 1442:1447 arg1 forms 1485:1489 possible active or inactive intermediate processive forms 1433:1489 possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1433:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 5 1 from site 1113:1116 arg1 position 1159:1166 position 59 1159:1169 position 59 1159:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 2 2 theme complete 327:334 arg1 sequences 343:351 the complete coding sequences 323:351 the complete coding sequences for human and mouse preprocathepsin B 323:389 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 5 3 theme single 1140:1145 arg1 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 7 4 from forms 1485:1489 arg1 regulation 1533:1542 the transcriptional regulation 1513:1542 the transcriptional regulation of the cathepsin B gene 1513:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 3 5 theme single 711:716 arg1 chain 718:722 single chain 711:722 single chain 711:722 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 6 6 theme sequences 1206:1214 arg1 analysis 1184:1191 Comparative analysis 1172:1191 Comparative analysis of the three sequences 1172:1214 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 3 7 theme preprocathepsin 532:546 arg1 B 548:548 preprocathepsin B 532:548 preprocathepsin B 532:548 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 4 8 theme B 815:815 arg1 sequences 817:825 procathepsin B sequences 802:825 procathepsin B sequences from three species (human, mouse, and rat) 802:868 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 3 9 theme mature 703:708 arg1 B 735:735 mature (single chain) cathepsin B 703:735 mature (single chain) cathepsin B 703:735 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 1 10 theme additional 155:164 arg1 functions 181:189 additional extralysosomal functions 155:189 additional extralysosomal functions 155:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 2 11 theme enzyme 296:301 arg1 mode 240:243 mode 240:243 mode of biosynthesis 240:259 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 11 theme enzyme 296:301 arg1 investigations 207:220 our investigations 203:220 our investigations on the structure 203:237 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 11 theme enzyme 296:301 arg1 sorting 280:286 intracellular sorting 266:286 intracellular sorting of this enzyme 266:301 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 3 12 dep follows 588:594 arg1 sequence 634:641 a 17-residue NH2-terminal prepeptide sequence 597:641 follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus 588:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 6 13 theme multistep 1269:1277 arg1 processing 1235:1244 processing 1235:1244 processing of procathepsin B 1235:1262 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 6 13 theme multistep 1269:1277 arg1 process 1279:1285 a multistep process 1267:1285 a multistep process 1267:1285 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 7 14 from identification 1415:1428 arg1 regulation 1533:1542 the transcriptional regulation 1513:1542 the transcriptional regulation of the cathepsin B gene 1513:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 6 15 theme Comparative 1172:1182 arg1 analysis 1184:1191 Comparative analysis 1172:1191 Comparative analysis of the three sequences 1172:1214 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 3 16 theme 6-residue 744:752 arg1 extension 754:762 a 6-residue extension 742:762 a 6-residue extension at the COOH terminus 742:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 6 17 theme procathepsin 1249:1260 arg1 B 1262:1262 procathepsin B 1249:1262 procathepsin B 1249:1262 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 7 18 theme gene 1563:1566 arg1 regulation 1533:1542 the transcriptional regulation 1513:1542 the transcriptional regulation of the cathepsin B gene 1513:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 2 19 theme biosynthesis 248:259 arg1 mode 240:243 mode 240:243 mode of biosynthesis 240:259 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 19 theme biosynthesis 248:259 arg1 investigations 207:220 our investigations 203:220 our investigations on the structure 203:237 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 19 theme biosynthesis 248:259 arg1 sorting 280:286 intracellular sorting 266:286 intracellular sorting of this enzyme 266:301 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 3 20 theme COOH 771:774 arg1 terminus 776:783 the COOH terminus 767:783 the COOH terminus 767:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 21 theme NH2-terminal 610:621 arg1 sequence 634:641 a 17-residue NH2-terminal prepeptide sequence 597:641 follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus 588:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 1 22 theme lysosomal 114:122 arg1 proteinase 130:139 a lysosomal thiol proteinase 112:139 a lysosomal thiol proteinase that may have additional extralysosomal functions 112:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 1 22 theme lysosomal 114:122 arg1 B 107:107 Cathepsin B 97:107 Cathepsin B 97:107 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 2 23 from sorting 280:286 arg1 structure 229:237 the structure 225:237 the structure 225:237 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 7 24 theme cathepsin 1551:1559 arg1 gene 1563:1566 the cathepsin B gene 1547:1566 the cathepsin B gene 1547:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 0 25 theme human 56:60 arg1 cDNAs 90:94 cloned human and mouse preprocathepsin B cDNAs 49:94 cloned human and mouse preprocathepsin B cDNAs 49:94 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 5 26 gly glycosylation 1099:1111 arg1 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 26 gly glycosylation 1099:1111 arg2 site 1113:1116 a potential glycosylation site 1087:1116 a potential glycosylation site 1087:1116 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 26 gly glycosylation 1099:1111 arg2 position 1159:1166 position 59 1159:1169 position 59 1159:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 26 gly glycosylation 1099:1111 arg2 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 0 27 theme mouse 66:70 arg1 cDNAs 90:94 cloned human and mouse preprocathepsin B cDNAs 49:94 cloned human and mouse preprocathepsin B cDNAs 49:94 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 4 28 from species 838:844 arg1 sequences 817:825 procathepsin B sequences 802:825 procathepsin B sequences from three species (human, mouse, and rat) 802:868 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 4 28 from species 838:844 arg1 comparison 788:797 A comparison 786:797 A comparison of procathepsin B sequences from three species (human, mouse, and rat) 786:868 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 7 29 theme transcriptional 1517:1531 arg1 regulation 1533:1542 the transcriptional regulation 1513:1542 the transcriptional regulation of the cathepsin B gene 1513:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 0 30 theme B 88:88 arg1 cDNAs 90:94 cloned human and mouse preprocathepsin B cDNAs 49:94 cloned human and mouse preprocathepsin B cDNAs 49:94 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 2 31 theme intracellular 266:278 arg1 sorting 280:286 intracellular sorting 266:286 intracellular sorting of this enzyme 266:301 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 32 theme preprocathepsin 373:387 arg1 B 389:389 preprocathepsin B 373:389 preprocathepsin B 373:389 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 33 theme human 426:430 arg1 hepatoma 432:439 human hepatoma and kidney phage libraries 426:466 hepatoma 432:439 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 34 from mode 240:243 arg1 structure 229:237 the structure 225:237 the structure 225:237 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 4 35 theme identity 975:982 arg1 minimum 951:957 a minimum 949:957 a minimum of 68% sequence identity 949:982 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 2 36 theme cDNA 400:403 arg1 clones 405:410 cDNA clones 400:410 cDNA clones isolated from human hepatoma and kidney phage libraries 400:466 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 0 37 theme predicted 15:23 arg1 sequences 36:44 predicted amino acid sequences 15:44 predicted amino acid sequences 15:44 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 1 38 theme Cathepsin 97:105 arg1 proteinase 130:139 a lysosomal thiol proteinase 112:139 a lysosomal thiol proteinase that may have additional extralysosomal functions 112:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 1 38 theme Cathepsin 97:105 arg1 B 107:107 Cathepsin B 97:107 Cathepsin B 97:107 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 2 39 theme phage 452:456 arg1 libraries 458:466 human hepatoma and kidney phage libraries 426:466 libraries 458:466 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 0 40 theme acid 31:34 arg1 sequences 36:44 predicted amino acid sequences 15:44 predicted amino acid sequences 15:44 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 3 41 dep mature 703:708 arg1 chain 718:722 single chain 711:722 single chain 711:722 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 7 42 theme forms 1485:1489 arg1 identification 1415:1428 the identification 1411:1428 the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1411:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 1 43 contain have 150:153 arg1 proteinase 130:139 a lysosomal thiol proteinase 112:139 a lysosomal thiol proteinase that may have additional extralysosomal functions 112:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 1 43 contain have 150:153 arg1 B 107:107 Cathepsin B 97:107 Cathepsin B 97:107 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 1 43 contain have 150:153 arg2 functions 181:189 additional extralysosomal functions 155:189 additional extralysosomal functions 155:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 5 44 theme glycosylation 1099:1111 arg1 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 44 theme glycosylation 1099:1111 arg1 site 1113:1116 a potential glycosylation site 1087:1116 a potential glycosylation site 1087:1116 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 7 45 theme intermediate 1461:1472 arg1 forms 1485:1489 possible active or inactive intermediate processive forms 1433:1489 possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1433:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 3 46 theme 62-residue 657:666 arg1 residues 691:698 254 residues 687:698 254 residues in mature (single chain) cathepsin B 687:735 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 46 theme 62-residue 657:666 arg1 region 679:684 a 62-residue propeptide region 655:684 a 62-residue propeptide region 655:684 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 4 47 dep species 838:844 arg1 mouse 854:858 mouse 854:858 mouse 854:858 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 4 47 dep species 838:844 arg1 rat 865:867 rat 865:867 rat 865:867 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 4 47 dep species 838:844 arg1 human 847:851 human 847:851 human 847:851 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 7 48 theme possible 1433:1440 arg1 forms 1485:1489 possible active or inactive intermediate processive forms 1433:1489 possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1433:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 6 49 theme intermediate 1322:1333 arg1 forms 1335:1339 enzymatically active intermediate forms 1301:1339 enzymatically active intermediate forms 1301:1339 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 2 50 theme coding 336:341 arg1 sequences 343:351 the complete coding sequences 323:351 the complete coding sequences for human and mouse preprocathepsin B 323:389 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 5 51 theme cysteine 1147:1154 arg1 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 51 theme cysteine 1147:1154 arg1 site 1113:1116 a potential glycosylation site 1087:1116 a potential glycosylation site 1087:1116 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 51 theme cysteine 1147:1154 arg1 presence 1126:1133 the presence 1122:1133 the presence of a single cysteine at position 59 1122:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 7 52 theme cDNA 1383:1386 arg1 clones 1388:1393 the cDNA clones 1379:1393 the cDNA clones 1379:1393 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 5 53 from presence 1126:1133 arg1 position 1159:1166 position 59 1159:1169 position 59 1159:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 3 54 theme primary 511:517 arg1 structure 519:527 the primary structure 507:527 the primary structure of preprocathepsin B 507:548 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 7 55 theme clones 1388:1393 arg1 availability 1363:1374 The availability 1359:1374 The availability of the cDNA clones 1359:1393 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 4 56 theme procathepsin 802:813 arg1 sequences 817:825 procathepsin B sequences 802:825 procathepsin B sequences from three species (human, mouse, and rat) 802:868 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 6 57 theme active 1315:1320 arg1 forms 1335:1339 enzymatically active intermediate forms 1301:1339 enzymatically active intermediate forms 1301:1339 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 1 58 theme extralysosomal 166:179 arg1 functions 181:189 additional extralysosomal functions 155:189 additional extralysosomal functions 155:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 4 59 theme sequences 817:825 arg1 comparison 788:797 A comparison 786:797 A comparison of procathepsin B sequences from three species (human, mouse, and rat) 786:868 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 3 60 theme B 548:548 arg1 structure 519:527 the primary structure 507:527 the primary structure of preprocathepsin B 507:548 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 61 theme amino 563:567 arg1 acids 569:573 339 amino acids 559:573 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus 559:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 62 theme cathepsin 725:733 arg1 B 735:735 mature (single chain) cathepsin B 703:735 mature (single chain) cathepsin B 703:735 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 5 63 theme conserved 1004:1012 arg1 sequences 1014:1022 two conserved sequences 1000:1022 two conserved sequences in the propeptide that may be functionally significant 1000:1077 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 3 64 from residues 691:698 arg1 B 735:735 mature (single chain) cathepsin B 703:735 mature (single chain) cathepsin B 703:735 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 5 65 from sequences 1014:1022 arg1 propeptide 1031:1040 the propeptide 1027:1040 the propeptide 1027:1040 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 6 66 theme B 1262:1262 arg1 processing 1235:1244 processing 1235:1244 processing of procathepsin B 1235:1262 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 6 66 theme B 1262:1262 arg1 process 1279:1285 a multistep process 1267:1285 a multistep process 1267:1285 Comparative analysis of the three sequences also suggests that processing of procathepsin B is a multistep process, during which enzymatically active intermediate forms may be generated. 3463996 4 67 from comparison 788:797 arg1 species 838:844 three species 832:844 three species (human, mouse, and rat) 832:868 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 7 68 theme B 1561:1561 arg1 gene 1563:1566 the cathepsin B gene 1547:1566 the cathepsin B gene 1547:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 5 69 from position 1159:1166 arg1 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 69 from position 1159:1166 arg1 site 1113:1116 a potential glycosylation site 1087:1116 a potential glycosylation site 1087:1116 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 69 from position 1159:1166 arg1 presence 1126:1133 the presence 1122:1133 the presence of a single cysteine at position 59 1122:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 3 70 theme 17-residue 599:608 arg1 sequence 634:641 a 17-residue NH2-terminal prepeptide sequence 597:641 follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus 588:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 1 71 theme thiol 124:128 arg1 proteinase 130:139 a lysosomal thiol proteinase 112:139 a lysosomal thiol proteinase that may have additional extralysosomal functions 112:189 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 1 71 theme thiol 124:128 arg1 B 107:107 Cathepsin B 97:107 Cathepsin B 97:107 Cathepsin B is a lysosomal thiol proteinase that may have additional extralysosomal functions. 3463996 0 72 theme preprocathepsin 72:86 arg1 cDNAs 90:94 cloned human and mouse preprocathepsin B cDNAs 49:94 cloned human and mouse preprocathepsin B cDNAs 49:94 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 0 73 theme cDNAs 90:94 arg1 sequences 36:44 predicted amino acid sequences 15:44 predicted amino acid sequences 15:44 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 0 73 theme cDNAs 90:94 arg1 Nucleotide 0:9 Nucleotide 0:9 Nucleotide 0:9 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 7 74 theme studies 1502:1508 arg1 identification 1415:1428 the identification 1411:1428 the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1411:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 2 75 from investigations 207:220 arg1 structure 229:237 the structure 225:237 the structure 225:237 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 4 76 theme 68 962:963 arg1 % 964:964 % 964:964 % 964:964 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 3 77 contain contains 550:557 arg2 acids 569:573 339 amino acids 559:573 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus 559:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 77 contain contains 550:557 arg1 structure 519:527 the primary structure 507:527 the primary structure of preprocathepsin B 507:548 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 0 78 theme amino 25:29 arg1 sequences 36:44 predicted amino acid sequences 15:44 predicted amino acid sequences 15:44 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 7 79 from studies 1502:1508 arg1 regulation 1533:1542 the transcriptional regulation 1513:1542 the transcriptional regulation of the cathepsin B gene 1513:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 2 80 attach isolated 412:419 arg2 clones 405:410 cDNA clones 400:410 cDNA clones isolated from human hepatoma and kidney phage libraries 400:466 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 80 attach isolated 412:419 arg1 libraries 458:466 human hepatoma and kidney phage libraries 426:466 libraries 458:466 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 2 80 attach isolated 412:419 arg1 hepatoma 432:439 human hepatoma and kidney phage libraries 426:466 hepatoma 432:439 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 4 81 theme sequence 966:973 arg1 identity 975:982 68% sequence identity 962:982 68% sequence identity 962:982 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 3463996 2 82 theme kidney 445:450 arg1 libraries 458:466 human hepatoma and kidney phage libraries 426:466 libraries 458:466 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 3 83 theme prepeptide 623:632 arg1 sequence 634:641 a 17-residue NH2-terminal prepeptide sequence 597:641 follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus 588:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 0 84 theme cloned 49:54 arg1 cDNAs 90:94 cloned human and mouse preprocathepsin B cDNAs 49:94 cloned human and mouse preprocathepsin B cDNAs 49:94 Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs. 3463996 5 85 theme potential 1089:1097 arg1 cysteine 1147:1154 a single cysteine 1138:1154 a single cysteine at position 59 1138:1169 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 5 85 theme potential 1089:1097 arg1 site 1113:1116 a potential glycosylation site 1087:1116 a potential glycosylation site 1087:1116 In particular, two conserved sequences in the propeptide that may be functionally significant include a potential glycosylation site and the presence of a single cysteine at position 59. 3463996 7 86 from regulation 1533:1542 arg1 identification 1415:1428 the identification 1411:1428 the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1411:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 3 87 theme propeptide 668:677 arg1 residues 691:698 254 residues 687:698 254 residues in mature (single chain) cathepsin B 687:735 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 87 theme propeptide 668:677 arg1 region 679:684 a 62-residue propeptide region 655:684 a 62-residue propeptide region 655:684 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 88 theme nucleotide 473:482 arg1 sequences 484:492 The nucleotide sequences 469:492 The nucleotide sequences 469:492 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 7 89 theme processive 1474:1483 arg1 forms 1485:1489 possible active or inactive intermediate processive forms 1433:1489 possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1433:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 3 90 from terminus 776:783 arg1 residues 691:698 254 residues 687:698 254 residues in mature (single chain) cathepsin B 687:735 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 90 from terminus 776:783 arg1 extension 754:762 a 6-residue extension 742:762 a 6-residue extension at the COOH terminus 742:783 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 3 90 from terminus 776:783 arg1 region 679:684 a 62-residue propeptide region 655:684 a 62-residue propeptide region 655:684 The nucleotide sequences predict that the primary structure of preprocathepsin B contains 339 amino acids organized as follows: a 17-residue NH2-terminal prepeptide sequence followed by a 62-residue propeptide region, 254 residues in mature (single chain) cathepsin B, and a 6-residue extension at the COOH terminus. 3463996 2 91 dep human 357:361 arg1 B 389:389 preprocathepsin B 373:389 preprocathepsin B 373:389 To further our investigations on the structure, mode of biosynthesis, and intracellular sorting of this enzyme, we have determined the complete coding sequences for human and mouse preprocathepsin B by using cDNA clones isolated from human hepatoma and kidney phage libraries. 3463996 7 92 theme inactive 1452:1459 arg1 forms 1485:1489 possible active or inactive intermediate processive forms 1433:1489 possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene 1433:1566 The availability of the cDNA clones will facilitate the identification of possible active or inactive intermediate processive forms as well as studies on the transcriptional regulation of the cathepsin B gene. 3463996 4 93 theme % 964:964 arg1 identity 975:982 68% sequence identity 962:982 68% sequence identity 962:982 A comparison of procathepsin B sequences from three species (human, mouse, and rat) reveals that the homology between the propeptides is relatively conserved with a minimum of 68% sequence identity. 15662415 4 0 dep elongated 502:510 arg1 curved 513:518 curved 513:518 curved 513:518 The hormone is bound in a hand-clasp fashion to an elongated, curved receptor. 15662415 3 1 theme 2.9-A-resolution 295:310 arg1 structure 312:320 the 2.9-A-resolution structure 291:320 the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)) 291:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 8 2 theme HB 980:981 arg1 complexes 984:992 The FSH-FSHR(HB) complexes 967:992 The FSH-FSHR(HB) complexes 967:992 The FSH-FSHR(HB) complexes form dimers in the crystal and at high concentrations in solution. 15662415 5 3 theme complex 558:564 arg1 large 569:573 large 569:573 large 569:573 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 3 theme complex 558:564 arg1 interface 541:549 The buried interface 530:549 The buried interface of the complex 530:564 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 3 theme complex 558:564 arg1 A2 582:583 2,600 A2 576:583 2,600 A2 576:583 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 3 4 theme human 363:367 arg1 FSH 369:371 human FSH 363:371 human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)) 363:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 0 5 from Structure 0:8 arg1 complex 51:57 complex 51:57 complex with its receptor 51:75 Structure of human follicle-stimulating hormone in complex with its receptor. 15662415 5 6 theme high 596:599 arg1 density 608:614 a high charge density 594:614 a high charge density 594:614 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 2 7 from receptor 188:195 arg1 surface 204:210 the surface 200:210 the surface of target cells 200:226 It acts through a G-protein-coupled receptor on the surface of target cells to stimulate testicular and ovarian functions. 15662415 5 8 theme charge 601:606 arg1 density 608:614 a high charge density 594:614 a high charge density 594:614 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 6 9 theme glycoprotein 648:659 arg1 hormones 661:668 all glycoprotein hormones 644:668 all glycoprotein hormones 644:668 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 3 10 theme complex 352:358 arg1 structure 312:320 the 2.9-A-resolution structure 291:320 the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)) 291:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 8 11 theme FSH-FSHR 971:978 arg1 complexes 984:992 The FSH-FSHR(HB) complexes 967:992 The FSH-FSHR(HB) complexes 967:992 The FSH-FSHR(HB) complexes form dimers in the crystal and at high concentrations in solution. 15662415 6 12 theme binding 716:722 arg1 specificity 724:734 binding specificity 716:734 binding specificity 716:734 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 5 13 theme buried 534:539 arg1 large 569:573 large 569:573 large 569:573 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 13 theme buried 534:539 arg1 interface 541:549 The buried interface 530:549 The buried interface of the complex 530:564 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 13 theme buried 534:539 arg1 A2 582:583 2,600 A2 576:583 2,600 A2 576:583 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 1 14 theme Follicle-stimulating 78:97 arg1 FSH 108:110 FSH 108:110 FSH 108:110 Follicle-stimulating hormone (FSH) is central to reproduction in mammals. 15662415 1 14 theme Follicle-stimulating 78:97 arg1 hormone 99:105 Follicle-stimulating hormone 78:105 Follicle-stimulating hormone (FSH) 78:111 Follicle-stimulating hormone (FSH) is central to reproduction in mammals. 15662415 6 15 theme hormone-specific 810:825 arg1 beta-subunits 827:839 hormone-specific beta-subunits 810:839 hormone-specific beta-subunits 810:839 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 1 16 from reproduction 127:138 arg1 mammals 143:149 mammals 143:149 mammals 143:149 Follicle-stimulating hormone (FSH) is central to reproduction in mammals. 15662415 3 17 theme receptor 430:437 arg1 domain 416:421 the extracellular hormone-binding domain 382:421 the extracellular hormone-binding domain of its receptor (FSHR(HB)) 382:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 3 17 theme receptor 430:437 arg1 receptor 430:437 its receptor 426:437 its receptor (FSHR(HB)) 426:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 3 18 theme extracellular 386:398 arg1 domain 416:421 the extracellular hormone-binding domain 382:421 the extracellular hormone-binding domain of its receptor (FSHR(HB)) 382:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 3 18 theme extracellular 386:398 arg1 receptor 430:437 its receptor 426:437 its receptor (FSHR(HB)) 426:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 0 19 theme follicle-stimulating 19:38 arg1 hormone 40:46 human follicle-stimulating hormone 13:46 human follicle-stimulating hormone 13:46 Structure of human follicle-stimulating hormone in complex with its receptor. 15662415 9 20 theme transmembrane 1092:1104 arg1 transduction 1113:1124 transmembrane signal transduction 1092:1124 transmembrane signal transduction 1092:1124 Such dimers may participate in transmembrane signal transduction. 15662415 7 21 theme concerted 870:878 arg1 change 895:900 a concerted conformational change 868:900 a concerted conformational change that affects protruding loops implicated in receptor activation 868:964 On binding, FSH undergoes a concerted conformational change that affects protruding loops implicated in receptor activation. 15662415 6 22 theme key 751:753 arg1 sites 767:771 key interaction sites 751:771 key interaction sites involving both the common alpha- and hormone-specific beta-subunits 751:839 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 0 23 theme human 13:17 arg1 hormone 40:46 human follicle-stimulating hormone 13:46 human follicle-stimulating hormone 13:46 Structure of human follicle-stimulating hormone in complex with its receptor. 15662415 2 24 theme G-protein-coupled 170:186 arg1 receptor 188:195 a G-protein-coupled receptor 168:195 a G-protein-coupled receptor on the surface of target cells 168:226 It acts through a G-protein-coupled receptor on the surface of target cells to stimulate testicular and ovarian functions. 15662415 9 25 theme Such 1061:1064 arg1 dimers 1066:1071 Such dimers 1061:1071 Such dimers 1061:1071 Such dimers may participate in transmembrane signal transduction. 15662415 0 26 theme hormone 40:46 arg1 Structure 0:8 Structure 0:8 Structure of human follicle-stimulating hormone in complex with its receptor. 0:76 Structure of human follicle-stimulating hormone in complex with its receptor. 15662415 8 27 theme high 1028:1031 arg1 concentrations 1033:1046 high concentrations 1028:1046 high concentrations in solution 1028:1058 The FSH-FSHR(HB) complexes form dimers in the crystal and at high concentrations in solution. 15662415 6 28 theme common 792:797 arg1 alpha- 799:804 common alpha- 792:804 common alpha- 792:804 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 2 29 theme cells 222:226 arg1 surface 204:210 the surface 200:210 the surface of target cells 200:226 It acts through a G-protein-coupled receptor on the surface of target cells to stimulate testicular and ovarian functions. 15662415 6 30 dep alpha- 799:804 arg1 the 788:790 the 788:790 the 788:790 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 3 31 theme hormone-binding 400:414 arg1 domain 416:421 the extracellular hormone-binding domain 382:421 the extracellular hormone-binding domain of its receptor (FSHR(HB)) 382:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 3 31 theme hormone-binding 400:414 arg1 receptor 430:437 its receptor 426:437 its receptor (FSHR(HB)) 426:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 3 32 gly deglycosylated 337:350 arg1 complex 352:358 a partially deglycosylated complex 325:358 a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)) 325:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 2 33 theme target 215:220 arg1 cells 222:226 target cells 215:226 target cells 215:226 It acts through a G-protein-coupled receptor on the surface of target cells to stimulate testicular and ovarian functions. 15662415 4 34 theme hand-clasp 477:486 arg1 fashion 488:494 a hand-clasp fashion 475:494 a hand-clasp fashion 475:494 The hormone is bound in a hand-clasp fashion to an elongated, curved receptor. 15662415 7 35 theme conformational 880:893 arg1 change 895:900 a concerted conformational change 868:900 a concerted conformational change that affects protruding loops implicated in receptor activation 868:964 On binding, FSH undergoes a concerted conformational change that affects protruding loops implicated in receptor activation. 15662415 5 36 dep large 569:573 arg1 large 569:573 large 569:573 large 569:573 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 36 dep large 569:573 arg1 A2 582:583 2,600 A2 576:583 2,600 A2 576:583 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 36 dep large 569:573 arg1 interface 541:549 The buried interface 530:549 The buried interface of the complex 530:564 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 4 37 theme elongated 502:510 arg1 receptor 520:527 an elongated, curved receptor 499:527 an elongated, curved receptor 499:527 The hormone is bound in a hand-clasp fashion to an elongated, curved receptor. 15662415 6 38 gly glycoprotein 648:659 arg1 glycoprotein 648:659 all glycoprotein hormones 644:668 all glycoprotein hormones 644:668 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 2 39 theme ovarian 256:262 arg1 functions 264:272 testicular and ovarian functions 241:272 testicular and ovarian functions 241:272 It acts through a G-protein-coupled receptor on the surface of target cells to stimulate testicular and ovarian functions. 15662415 8 40 from concentrations 1033:1046 arg1 solution 1051:1058 solution 1051:1058 solution 1051:1058 The FSH-FSHR(HB) complexes form dimers in the crystal and at high concentrations in solution. 15662415 9 41 theme signal 1106:1111 arg1 transduction 1113:1124 transmembrane signal transduction 1092:1124 transmembrane signal transduction 1092:1124 Such dimers may participate in transmembrane signal transduction. 15662415 7 42 theme receptor 946:953 arg1 activation 955:964 receptor activation 946:964 receptor activation 946:964 On binding, FSH undergoes a concerted conformational change that affects protruding loops implicated in receptor activation. 15662415 6 43 theme interaction 755:765 arg1 sites 767:771 key interaction sites 751:771 key interaction sites involving both the common alpha- and hormone-specific beta-subunits 751:839 Our analysis suggests that all glycoprotein hormones bind to their receptors in this mode and that binding specificity is mediated by key interaction sites involving both the common alpha- and hormone-specific beta-subunits. 15662415 2 44 theme testicular 241:250 arg1 functions 264:272 testicular and ovarian functions 241:272 testicular and ovarian functions 241:272 It acts through a G-protein-coupled receptor on the surface of target cells to stimulate testicular and ovarian functions. 15662415 0 45 with complex 51:57 arg1 receptor 68:75 its receptor 64:75 its receptor 64:75 Structure of human follicle-stimulating hormone in complex with its receptor. 15662415 7 46 theme protruding 915:924 arg1 loops 926:930 protruding loops 915:930 protruding loops implicated in receptor activation 915:964 On binding, FSH undergoes a concerted conformational change that affects protruding loops implicated in receptor activation. 15662415 3 47 theme deglycosylated 337:350 arg1 complex 352:358 a partially deglycosylated complex 325:358 a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)) 325:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 15662415 5 48 contain has 590:592 arg2 density 608:614 a high charge density 594:614 a high charge density 594:614 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 48 contain has 590:592 arg1 large 569:573 large 569:573 large 569:573 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 48 contain has 590:592 arg1 interface 541:549 The buried interface 530:549 The buried interface of the complex 530:564 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 5 48 contain has 590:592 arg1 A2 582:583 2,600 A2 576:583 2,600 A2 576:583 The buried interface of the complex is large (2,600 A2) and has a high charge density. 15662415 3 49 theme FSH 369:371 arg1 complex 352:358 a partially deglycosylated complex 325:358 a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)) 325:448 We present here the 2.9-A-resolution structure of a partially deglycosylated complex of human FSH bound to the extracellular hormone-binding domain of its receptor (FSHR(HB)). 22809326 11 0 theme glycosylated 1313:1324 arg1 regions 1334:1340 the glycosylated luminal regions 1309:1340 the glycosylated luminal regions of LAMP-1 and LAMP-2 1309:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 9 1 theme β-prism 1106:1112 arg1 fold 1114:1117 a novel β-prism fold 1098:1117 a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond 1098:1191 The structure represents a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond. 22809326 11 2 theme LAMP-2 1356:1361 arg1 regions 1334:1340 the glycosylated luminal regions 1309:1340 the glycosylated luminal regions of LAMP-1 and LAMP-2 1309:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 12 3 dep 12 1552:1553 arg1 to 1549:1550 to 1549:1550 to 1549:1550 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 5 4 theme abundant 623:630 arg1 proteins 632:639 The abundant proteins 619:639 The abundant proteins LAMP-1 and LAMP-2 619:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 4 theme abundant 623:630 arg1 LAMP-2 652:657 LAMP-2 652:657 LAMP-2 652:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 4 theme abundant 623:630 arg1 constituents 669:680 major constituents 663:680 major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx' 663:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 4 theme abundant 623:630 arg1 LAMP-1 641:646 LAMP-1 641:646 LAMP-1 641:646 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 3 5 theme LAMP-2 500:505 arg1 A 515:515 LAMP-2 isoform A 500:515 LAMP-2 isoform A 500:515 LAMP-2 isoform A acts as a receptor in chaperone-mediated autophagy. 22809326 3 5 theme LAMP-2 500:505 arg1 receptor 527:534 a receptor 525:534 a receptor in chaperone-mediated autophagy 525:566 LAMP-2 isoform A acts as a receptor in chaperone-mediated autophagy. 22809326 11 6 theme LAMP-1 1345:1350 arg1 regions 1334:1340 the glycosylated luminal regions 1309:1340 the glycosylated luminal regions of LAMP-1 and LAMP-2 1309:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 1 7 theme lysosome-associated 160:178 arg1 LAMP 199:202 LAMP 199:202 LAMP 199:202 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 7 theme lysosome-associated 160:178 arg1 proteins 189:196 lysosome-associated membrane proteins 160:196 lysosome-associated membrane proteins (LAMP) 160:203 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 13 8 theme β-prism 1702:1708 arg1 fold 1710:1713 a previously unknown β-prism fold 1681:1713 a previously unknown β-prism fold 1681:1713 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 9 9 theme bent 1142:1145 arg1 β-sheets 1133:1140 two β-sheets 1129:1140 two β-sheets bent by β-bulges 1129:1157 The structure represents a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond. 22809326 12 10 gly glycoprotein 1521:1532 arg1 glycoprotein 1521:1532 the lysosomal glycoprotein coat 1507:1537 the lysosomal glycoprotein coat of only 5 to 12 nm 1507:1556 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 1 11 theme proteins 189:196 arg1 family 150:155 The family 146:155 The family of lysosome-associated membrane proteins (LAMP) 146:203 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 5 12 theme major 663:667 arg1 proteins 632:639 The abundant proteins 619:639 The abundant proteins LAMP-1 and LAMP-2 619:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 12 theme major 663:667 arg1 constituents 669:680 major constituents 663:680 major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx' 663:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 11 13 theme compact 1398:1404 arg1 conformation 1406:1417 a compact conformation 1396:1417 a compact conformation 1396:1417 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 4 14 theme Danon 604:608 arg1 disease 610:616 the fatal Danon disease 594:616 the fatal Danon disease 594:616 LAMP-2 deficiency causes the fatal Danon disease. 22809326 3 15 theme chaperone-mediated 539:556 arg1 autophagy 558:566 chaperone-mediated autophagy 539:566 chaperone-mediated autophagy 539:566 LAMP-2 isoform A acts as a receptor in chaperone-mediated autophagy. 22809326 7 16 theme domain 941:946 arg1 structure 914:922 The crystal structure 902:922 The crystal structure of the conserved domain of human DC-LAMP 902:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 12 17 theme coat 1534:1537 arg1 thickness 1494:1502 the thickness 1490:1502 the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm 1490:1556 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 12 18 theme lysosomal 1511:1519 arg1 coat 1534:1537 the lysosomal glycoprotein coat 1507:1537 the lysosomal glycoprotein coat of only 5 to 12 nm 1507:1556 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 1 19 dep multifunctional 219:233 arg1 ubiquitous 236:245 ubiquitous 236:245 ubiquitous 236:245 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 11 20 theme Computational 1285:1297 arg1 models 1299:1304 Computational models 1285:1304 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 1285:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 5 21 from inside 724:729 arg1 proteins 632:639 The abundant proteins 619:639 The abundant proteins LAMP-1 and LAMP-2 619:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 21 from inside 724:729 arg1 constituents 669:680 major constituents 663:680 major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx' 663:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 6 22 theme acids 861:865 arg1 domain 834:839 a conserved domain 822:839 a conserved domain of 150 to 200 amino acids with two disulfide bonds 822:890 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 6 22 theme acids 861:865 arg1 acids 861:865 150 to 200 amino acids 844:865 150 to 200 amino acids 844:865 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 7 23 theme DC-LAMP 957:963 arg1 domain 941:946 the conserved domain 927:946 the conserved domain of human DC-LAMP 927:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 7 23 theme DC-LAMP 957:963 arg1 DC-LAMP 957:963 human DC-LAMP 951:963 human DC-LAMP 951:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 14 24 theme computational 1796:1808 arg1 models 1810:1815 computational models 1796:1815 computational models of the LAMP-1 and LAMP-2 luminal regions 1796:1856 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 11 25 theme close 1422:1426 arg1 proximity 1428:1436 close proximity 1422:1436 close proximity to the lysosomal membrane 1422:1462 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 0 26 theme DC 49:50 arg1 protein 82:88 the DC lysosomal associated membrane protein 45:88 the DC lysosomal associated membrane protein 45:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 0 27 theme associated 62:71 arg1 protein 82:88 the DC lysosomal associated membrane protein 45:88 the DC lysosomal associated membrane protein 45:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 14 28 theme coat 1774:1777 arg1 structure 1734:1742 the structure 1730:1742 the structure of the lysosomal glycoprotein coat 1730:1777 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 13 29 theme lysosome-associated 1637:1655 arg1 proteins 1666:1673 lysosome-associated membrane proteins 1637:1673 lysosome-associated membrane proteins 1637:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 11 30 theme lysosomal 1445:1453 arg1 membrane 1455:1462 the lysosomal membrane 1441:1462 the lysosomal membrane 1441:1462 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 0 31 theme protein 82:88 arg1 domain 35:40 the conserved domain 21:40 the conserved domain of the DC lysosomal associated membrane protein 21:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 0 31 theme protein 82:88 arg1 protein 82:88 the DC lysosomal associated membrane protein 45:88 the DC lysosomal associated membrane protein 45:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 5 32 dep proteins 632:639 arg1 proteins 632:639 The abundant proteins 619:639 The abundant proteins LAMP-1 and LAMP-2 619:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 32 dep proteins 632:639 arg1 LAMP-2 652:657 LAMP-2 652:657 LAMP-2 652:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 32 dep proteins 632:639 arg1 LAMP-1 641:646 LAMP-1 641:646 LAMP-1 641:646 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 33 theme present 709:715 arg1 proteins 632:639 The abundant proteins 619:639 The abundant proteins LAMP-1 and LAMP-2 619:657 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 33 theme present 709:715 arg1 constituents 669:680 major constituents 663:680 major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx' 663:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 13 34 theme conserved 1609:1617 arg1 proteins 1666:1673 lysosome-associated membrane proteins 1637:1673 lysosome-associated membrane proteins 1637:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 13 34 theme conserved 1609:1617 arg1 domain 1627:1632 The conserved luminal domain 1605:1632 The conserved luminal domain of lysosome-associated membrane proteins 1605:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 10 35 theme Flexible 1194:1201 arg1 loops 1203:1207 Flexible loops 1194:1207 Flexible loops 1194:1207 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 0 36 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 0:132 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 14 37 theme glycoprotein 1761:1772 arg1 coat 1774:1777 the lysosomal glycoprotein coat 1747:1777 the lysosomal glycoprotein coat 1747:1777 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 11 38 gly glycosylated 1313:1324 arg1 regions 1334:1340 the glycosylated luminal regions 1309:1340 the glycosylated luminal regions of LAMP-1 and LAMP-2 1309:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 3 39 from receptor 527:534 arg1 autophagy 558:566 chaperone-mediated autophagy 539:566 chaperone-mediated autophagy 539:566 LAMP-2 isoform A acts as a receptor in chaperone-mediated autophagy. 22809326 8 40 theme first 987:991 arg1 structure 1009:1017 the first high-resolution structure 983:1017 the first high-resolution structure of a heavily glycosylated lysosomal membrane protein 983:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 8 40 theme first 987:991 arg1 It 977:978 It 977:978 It 977:978 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 2 41 theme processes 427:435 arg1 multitude 405:413 a multitude 403:413 a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging 403:497 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 10 42 theme hydrophobic 1215:1225 arg1 pocket 1227:1232 a hydrophobic pocket 1213:1232 a hydrophobic pocket 1213:1232 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 5 43 theme lysosomal 738:746 arg1 membrane 748:755 the lysosomal membrane 734:755 the lysosomal membrane 734:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 0 44 theme conserved 25:33 arg1 domain 35:40 the conserved domain 21:40 the conserved domain of the DC lysosomal associated membrane protein 21:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 0 44 theme conserved 25:33 arg1 protein 82:88 the DC lysosomal associated membrane protein 45:88 the DC lysosomal associated membrane protein 45:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 10 45 theme molecular 1262:1270 arg1 interaction 1272:1282 molecular interaction 1262:1282 molecular interaction 1262:1282 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 6 46 theme LAMP 790:793 arg1 family 795:800 The LAMP family 786:800 The LAMP family 786:800 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 8 47 theme glycosylated 1032:1043 arg1 protein 1064:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 11 48 theme luminal 1326:1332 arg1 regions 1334:1340 the glycosylated luminal regions 1309:1340 the glycosylated luminal regions of LAMP-1 and LAMP-2 1309:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22809326 8 49 theme membrane 1055:1062 arg1 protein 1064:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 6 50 theme amino 855:859 arg1 acids 861:865 150 to 200 amino acids 844:865 150 to 200 amino acids 844:865 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 4 51 theme LAMP-2 569:574 arg1 deficiency 576:585 LAMP-2 deficiency 569:585 LAMP-2 deficiency 569:585 LAMP-2 deficiency causes the fatal Danon disease. 22809326 9 52 theme novel 1100:1104 arg1 fold 1114:1117 a novel β-prism fold 1098:1117 a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond 1098:1191 The structure represents a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond. 22809326 5 53 from constituents 669:680 arg1 inside 724:729 the inside 720:729 the inside of the lysosomal membrane 720:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 6 54 with domain 834:839 arg1 bonds 886:890 two disulfide bonds 872:890 two disulfide bonds 872:890 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 6 55 dep 200 851:853 arg1 to 848:849 to 848:849 to 848:849 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 6 56 theme conserved 824:832 arg1 domain 834:839 a conserved domain 822:839 a conserved domain of 150 to 200 amino acids with two disulfide bonds 822:890 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 6 56 theme conserved 824:832 arg1 acids 861:865 150 to 200 amino acids 844:865 150 to 200 amino acids 844:865 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 13 57 theme unknown 1694:1700 arg1 fold 1710:1713 a previously unknown β-prism fold 1681:1713 a previously unknown β-prism fold 1681:1713 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 1 58 theme membrane 180:187 arg1 LAMP 199:202 LAMP 199:202 LAMP 199:202 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 58 theme membrane 180:187 arg1 proteins 189:196 lysosome-associated membrane proteins 160:196 lysosome-associated membrane proteins (LAMP) 160:203 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 0 59 theme lysosomal 112:120 arg1 glycocalyx 122:131 the lysosomal glycocalyx 108:131 the lysosomal glycocalyx 108:131 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 3 60 theme isoform 507:513 arg1 A 515:515 LAMP-2 isoform A 500:515 LAMP-2 isoform A 500:515 LAMP-2 isoform A acts as a receptor in chaperone-mediated autophagy. 22809326 3 60 theme isoform 507:513 arg1 receptor 527:534 a receptor 525:534 a receptor in chaperone-mediated autophagy 525:566 LAMP-2 isoform A acts as a receptor in chaperone-mediated autophagy. 22809326 14 61 gly glycoprotein 1761:1772 arg1 glycoprotein 1761:1772 the lysosomal glycoprotein coat 1747:1777 the lysosomal glycoprotein coat 1747:1777 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 14 62 theme luminal 1842:1848 arg1 regions 1850:1856 the LAMP-1 and LAMP-2 luminal regions 1820:1856 the LAMP-1 and LAMP-2 luminal regions 1820:1856 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 13 63 theme proteins 1666:1673 arg1 proteins 1666:1673 lysosome-associated membrane proteins 1637:1673 lysosome-associated membrane proteins 1637:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 13 63 theme proteins 1666:1673 arg1 domain 1627:1632 The conserved luminal domain 1605:1632 The conserved luminal domain of lysosome-associated membrane proteins 1605:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 5 64 from present 709:715 arg1 inside 724:729 the inside 720:729 the inside of the lysosomal membrane 720:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 14 65 theme regions 1850:1856 arg1 models 1810:1815 computational models 1796:1815 computational models of the LAMP-1 and LAMP-2 luminal regions 1796:1856 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 6 66 theme disulfide 876:884 arg1 bonds 886:890 two disulfide bonds 872:890 two disulfide bonds 872:890 The LAMP family is characterized by a conserved domain of 150 to 200 amino acids with two disulfide bonds. 22809326 4 67 theme fatal 598:602 arg1 disease 610:616 the fatal Danon disease 594:616 the fatal Danon disease 594:616 LAMP-2 deficiency causes the fatal Danon disease. 22809326 7 68 theme human 951:955 arg1 DC-LAMP 957:963 human DC-LAMP 951:963 human DC-LAMP 951:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 9 69 theme disulfide 1178:1186 arg1 bond 1188:1191 a disulfide bond 1176:1191 a disulfide bond 1176:1191 The structure represents a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond. 22809326 7 70 theme conserved 931:939 arg1 domain 941:946 the conserved domain 927:946 the conserved domain of human DC-LAMP 927:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 7 70 theme conserved 931:939 arg1 DC-LAMP 957:963 human DC-LAMP 951:963 human DC-LAMP 951:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 0 71 theme lysosomal 52:60 arg1 protein 82:88 the DC lysosomal associated membrane protein 45:88 the DC lysosomal associated membrane protein 45:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 9 72 dep fold 1114:1117 arg1 connected 1163:1171 connected 1163:1171 connected by a disulfide bond 1163:1191 The structure represents a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond. 22809326 9 72 dep fold 1114:1117 arg1 formed 1119:1124 formed 1119:1124 formed by two β-sheets bent by β-bulges 1119:1157 The structure represents a novel β-prism fold formed by two β-sheets bent by β-bulges and connected by a disulfide bond. 22809326 5 73 theme glycoconjugate 689:702 arg1 present 709:715 the glycoconjugate coat present 685:715 the glycoconjugate coat present on the inside of the lysosomal membrane 685:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 73 theme glycoconjugate 689:702 arg1 glycocalyx 773:782 the 'lysosomal glycocalyx' 758:783 the 'lysosomal glycocalyx' 758:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 0 74 theme membrane 73:80 arg1 protein 82:88 the DC lysosomal associated membrane protein 45:88 the DC lysosomal associated membrane protein 45:88 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 7 75 theme crystal 906:912 arg1 structure 914:922 The crystal structure 902:922 The crystal structure of the conserved domain of human DC-LAMP 902:963 RESULTS: The crystal structure of the conserved domain of human DC-LAMP was solved. 22809326 2 76 theme lipid 473:477 arg1 transport 479:487 lipid transport 473:487 lipid transport 473:487 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 13 77 theme membrane 1657:1664 arg1 proteins 1666:1673 lysosome-associated membrane proteins 1637:1673 lysosome-associated membrane proteins 1637:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 5 78 theme coat 704:707 arg1 present 709:715 the glycoconjugate coat present 685:715 the glycoconjugate coat present on the inside of the lysosomal membrane 685:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 78 theme coat 704:707 arg1 glycocalyx 773:782 the 'lysosomal glycocalyx' 758:783 the 'lysosomal glycocalyx' 758:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 14 79 theme LAMP-2 1835:1840 arg1 regions 1850:1856 the LAMP-1 and LAMP-2 luminal regions 1820:1856 the LAMP-1 and LAMP-2 luminal regions 1820:1856 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 12 80 theme electron 1572:1579 arg1 microscopy 1581:1590 electron microscopy 1572:1590 electron microscopy 1572:1590 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 1 81 dep proteins 293:300 arg1 DC-LAMP 302:308 DC-LAMP 302:308 DC-LAMP (LAMP-3) 302:317 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 81 dep proteins 293:300 arg1 CD68 366:369 CD68 366:369 CD68 366:369 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 81 dep proteins 293:300 arg1 LAMP-3 311:316 LAMP-3 311:316 LAMP-3 311:316 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 81 dep proteins 293:300 arg1 BAD-LAMP 320:327 BAD-LAMP 320:327 BAD-LAMP (UNC-46, C20orf103) 320:347 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 81 dep proteins 293:300 arg1 macrosialin 353:363 macrosialin 353:363 macrosialin (CD68) 353:370 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 81 dep proteins 293:300 arg1 proteins 293:300 the cell type-specific proteins 270:300 the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68) 270:370 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 82 dep BAD-LAMP 320:327 arg1 C20orf103 338:346 C20orf103 338:346 C20orf103 338:346 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 13 83 theme luminal 1619:1625 arg1 proteins 1666:1673 lysosome-associated membrane proteins 1637:1673 lysosome-associated membrane proteins 1637:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 13 83 theme luminal 1619:1625 arg1 domain 1627:1632 The conserved luminal domain 1605:1632 The conserved luminal domain of lysosome-associated membrane proteins 1605:1673 CONCLUSION: The conserved luminal domain of lysosome-associated membrane proteins forms a previously unknown β-prism fold. 22809326 10 84 theme possible 1244:1251 arg1 sites 1253:1257 possible sites 1244:1257 possible sites of molecular interaction 1244:1282 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 10 84 theme possible 1244:1251 arg1 interaction 1272:1282 molecular interaction 1262:1282 molecular interaction 1262:1282 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 5 85 theme lysosomal 763:771 arg1 present 709:715 the glycoconjugate coat present 685:715 the glycoconjugate coat present on the inside of the lysosomal membrane 685:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 5 85 theme lysosomal 763:771 arg1 glycocalyx 773:782 the 'lysosomal glycocalyx' 758:783 the 'lysosomal glycocalyx' 758:783 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 14 86 theme lysosomal 1751:1759 arg1 coat 1774:1777 the lysosomal glycoprotein coat 1747:1777 the lysosomal glycoprotein coat 1747:1777 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 8 87 theme high-resolution 993:1007 arg1 structure 1009:1017 the first high-resolution structure 983:1017 the first high-resolution structure of a heavily glycosylated lysosomal membrane protein 983:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 8 87 theme high-resolution 993:1007 arg1 It 977:978 It 977:978 It 977:978 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 12 88 theme nm 1555:1556 arg1 coat 1534:1537 the lysosomal glycoprotein coat 1507:1537 the lysosomal glycoprotein coat of only 5 to 12 nm 1507:1556 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 8 89 gly glycosylated 1032:1043 arg1 protein 1064:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 0 90 theme domain 35:40 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 0:132 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 1 91 theme type-specific 279:291 arg1 BAD-LAMP 320:327 BAD-LAMP 320:327 BAD-LAMP (UNC-46, C20orf103) 320:347 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 91 theme type-specific 279:291 arg1 DC-LAMP 302:308 DC-LAMP 302:308 DC-LAMP (LAMP-3) 302:317 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 91 theme type-specific 279:291 arg1 macrosialin 353:363 macrosialin 353:363 macrosialin (CD68) 353:370 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 1 91 theme type-specific 279:291 arg1 proteins 293:300 the cell type-specific proteins 270:300 the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68) 270:370 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 2 92 theme cellular 418:425 arg1 phagocytosis 448:459 phagocytosis 448:459 phagocytosis 448:459 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 2 92 theme cellular 418:425 arg1 transport 479:487 lipid transport 473:487 lipid transport 473:487 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 2 92 theme cellular 418:425 arg1 aging 493:497 aging 493:497 aging 493:497 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 2 92 theme cellular 418:425 arg1 autophagy 462:470 autophagy 462:470 autophagy 462:470 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 2 92 theme cellular 418:425 arg1 processes 427:435 cellular processes 418:435 cellular processes 418:435 LAMPs have been implicated in a multitude of cellular processes, including phagocytosis, autophagy, lipid transport and aging. 22809326 10 93 theme interaction 1272:1282 arg1 sites 1253:1257 possible sites 1244:1257 possible sites of molecular interaction 1244:1282 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 10 93 theme interaction 1272:1282 arg1 interaction 1272:1282 molecular interaction 1262:1282 molecular interaction 1262:1282 Flexible loops and a hydrophobic pocket represent possible sites of molecular interaction. 22809326 5 94 theme membrane 748:755 arg1 inside 724:729 the inside 720:729 the inside of the lysosomal membrane 720:755 The abundant proteins LAMP-1 and LAMP-2 are major constituents of the glycoconjugate coat present on the inside of the lysosomal membrane, the 'lysosomal glycocalyx'. 22809326 12 95 theme glycoprotein 1521:1532 arg1 coat 1534:1537 the lysosomal glycoprotein coat 1507:1537 the lysosomal glycoprotein coat of only 5 to 12 nm 1507:1556 The models correspond to the thickness of the lysosomal glycoprotein coat of only 5 to 12 nm, according to electron microscopy. 22809326 8 96 theme lysosomal 1045:1053 arg1 protein 1064:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 a heavily glycosylated lysosomal membrane protein 1022:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 1 97 theme multifunctional 219:233 arg1 LAMP-1 247:252 the multifunctional, ubiquitous LAMP-1 215:252 the multifunctional, ubiquitous LAMP-1 215:252 BACKGROUND: The family of lysosome-associated membrane proteins (LAMP) comprises the multifunctional, ubiquitous LAMP-1 and LAMP-2, and the cell type-specific proteins DC-LAMP (LAMP-3), BAD-LAMP (UNC-46, C20orf103) and macrosialin (CD68). 22809326 8 98 theme protein 1064:1070 arg1 structure 1009:1017 the first high-resolution structure 983:1017 the first high-resolution structure of a heavily glycosylated lysosomal membrane protein 983:1070 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 8 98 theme protein 1064:1070 arg1 It 977:978 It 977:978 It 977:978 It is the first high-resolution structure of a heavily glycosylated lysosomal membrane protein. 22809326 14 99 theme LAMP-1 1824:1829 arg1 regions 1850:1856 the LAMP-1 and LAMP-2 luminal regions 1820:1856 the LAMP-1 and LAMP-2 luminal regions 1820:1856 Insights into the structure of the lysosomal glycoprotein coat were obtained by computational models of the LAMP-1 and LAMP-2 luminal regions. 22809326 0 100 dep structure 8:16 arg1 implications 91:102 implications 91:102 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 0:132 Crystal structure of the conserved domain of the DC lysosomal associated membrane protein: implications for the lysosomal glycocalyx. 22809326 11 101 theme regions 1334:1340 arg1 models 1299:1304 Computational models 1285:1304 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 1285:1361 Computational models of the glycosylated luminal regions of LAMP-1 and LAMP-2 indicate that the proteins adopt a compact conformation in close proximity to the lysosomal membrane. 22601780 10 0 contain having 1088:1093 arg2 motif 1103:1107 the RGD motif 1095:1107 the RGD motif 1095:1107 Mutagenesis data confirm the importance of having the RGD motif at the tip of the RGD finger. 22601780 10 0 contain having 1088:1093 arg1 importance 1074:1083 the importance 1070:1083 the importance of having the RGD motif at the tip of the RGD finger 1070:1136 Mutagenesis data confirm the importance of having the RGD motif at the tip of the RGD finger. 22601780 5 1 theme domains 587:593 arg1 structure 564:572 the crystal structure 552:572 the crystal structure of its 3 EGF domains 552:593 We report the crystal structure of its 3 EGF domains. 22601780 11 2 theme EGF 1161:1163 arg1 sequences 1172:1180 EGF domain sequences 1161:1180 EGF domain sequences 1161:1180 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 8 3 link O-linked 882:889 arg1 glycans 891:897 Two unique O-linked glycans 871:897 Two unique O-linked glycans 871:897 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 0 4 theme critical 55:62 arg1 finger 8:13 The RGD finger 0:13 The RGD finger of Del-1 0:22 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 0 4 theme critical 55:62 arg1 feature 47:53 a unique structural feature 27:53 a unique structural feature critical for integrin binding 27:83 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 5 5 theme crystal 556:562 arg1 structure 564:572 the crystal structure 552:572 the crystal structure of its 3 EGF domains 552:593 We report the crystal structure of its 3 EGF domains. 22601780 3 6 contain contains 344:351 arg2 repeats 390:396 3 epidermal growth factor (EGF)-like repeats 353:396 3 epidermal growth factor (EGF)-like repeats 353:396 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 3 6 contain contains 344:351 arg1 Del-1 338:342 Del-1 338:342 Del-1 338:342 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 3 6 contain contains 344:351 arg2 domains 419:425 2 discoidin-like domains 402:425 2 discoidin-like domains 402:425 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 9 7 theme integrin 1009:1016 arg1 binding 1018:1024 integrin binding 1009:1024 integrin binding of the RGD finger 1009:1042 These structural features favor integrin binding of the RGD finger. 22601780 8 8 theme RGD 966:968 arg1 motif 970:974 the RGD motif 962:974 the RGD motif 962:974 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 9 9 theme structural 983:992 arg1 features 994:1001 These structural features 977:1001 These structural features 977:1001 These structural features favor integrin binding of the RGD finger. 22601780 6 10 theme long 658:661 arg1 finger 695:700 the RGD finger 687:700 dubbed the RGD finger 680:700 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 6 10 theme long 658:661 arg1 loop 674:677 a long protruding loop 656:677 a long protruding loop 656:677 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 11 11 theme Del-1 1277:1281 arg1 Del-1 1277:1281 Del-1 1277:1281 Del-1 1277:1281 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 11 theme Del-1 1277:1281 arg1 globule-EGF 1310:1320 its homologue milk fat globule-EGF 8 1287:1322 its homologue milk fat globule-EGF 8 1287:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 11 theme Del-1 1277:1281 arg1 domain 1267:1272 the EGF domain 1259:1272 the EGF domain of Del-1 and its homologue milk fat globule-EGF 8 1259:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 0 12 theme integrin 68:75 arg1 binding 77:83 integrin binding 68:83 integrin binding 68:83 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 10 13 theme RGD 1127:1129 arg1 finger 1131:1136 the RGD finger 1123:1136 the RGD finger 1123:1136 Mutagenesis data confirm the importance of having the RGD motif at the tip of the RGD finger. 22601780 7 14 theme binding 779:785 arg1 site 787:790 an interdomain calcium ion binding site 752:790 an interdomain calcium ion binding site 752:790 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 6 15 theme RGD 600:602 arg1 motif 604:608 The RGD motif 596:608 The RGD motif of EGF2 596:616 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 10 16 theme RGD 1099:1101 arg1 motif 1103:1107 the RGD motif 1095:1107 the RGD motif 1095:1107 Mutagenesis data confirm the importance of having the RGD motif at the tip of the RGD finger. 22601780 8 17 link N-linked 905:912 arg1 glycan 914:919 1 N-linked glycan 903:919 1 N-linked glycan 903:919 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 1 18 theme endothelial 100:110 arg1 locus-1 117:123 Developmental endothelial cell locus-1 86:123 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 1 18 theme endothelial 100:110 arg1 Del-1 126:130 Del-1 126:130 Del-1 126:130 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 8 19 theme unique 875:880 arg1 glycans 891:897 Two unique O-linked glycans 871:897 Two unique O-linked glycans 871:897 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 11 20 theme milk 1301:1304 arg1 globule-EGF 1310:1320 its homologue milk fat globule-EGF 8 1287:1322 its homologue milk fat globule-EGF 8 1287:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 2 21 dep β 332:332 arg1 V 330:330 V 330:330 V 330:330 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 3 22 theme growth 365:370 arg1 factor 372:377 3 epidermal growth factor 353:377 3 epidermal growth factor 353:377 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 7 23 theme calcium 767:773 arg1 site 787:790 an interdomain calcium ion binding site 752:790 an interdomain calcium ion binding site 752:790 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 2 24 theme vascular 241:248 arg1 remodeling 250:259 vascular remodeling 241:259 vascular remodeling 241:259 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 0 25 theme RGD 4:6 arg1 finger 8:13 The RGD finger 0:13 The RGD finger of Del-1 0:22 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 0 25 theme RGD 4:6 arg1 feature 47:53 a unique structural feature 27:53 a unique structural feature critical for integrin binding 27:83 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 8 26 theme EGF2 952:955 arg1 side 944:947 the opposite side 931:947 the opposite side of EGF2 from the RGD motif 931:974 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 7 27 theme ion 775:777 arg1 site 787:790 an interdomain calcium ion binding site 752:790 an interdomain calcium ion binding site 752:790 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 11 28 theme EGF 1263:1265 arg1 Del-1 1277:1281 Del-1 1277:1281 Del-1 1277:1281 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 28 theme EGF 1263:1265 arg1 globule-EGF 1310:1320 its homologue milk fat globule-EGF 8 1287:1322 its homologue milk fat globule-EGF 8 1287:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 28 theme EGF 1263:1265 arg1 domain 1267:1272 the EGF domain 1259:1272 the EGF domain of Del-1 and its homologue milk fat globule-EGF 8 1259:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 1 29 theme endothelial 161:171 arg1 cells 173:177 endothelial cells 161:177 endothelial cells 161:177 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 4 30 theme EGF 469:471 arg1 EGF2 481:484 EGF2 481:484 EGF2 481:484 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 4 30 theme EGF 469:471 arg1 domain 473:478 the second EGF domain 458:478 the second EGF domain (EGF2) 458:485 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 8 31 theme O-linked 882:889 arg1 glycans 891:897 Two unique O-linked glycans 871:897 Two unique O-linked glycans 871:897 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 9 32 theme finger 1037:1042 arg1 binding 1018:1024 integrin binding 1009:1024 integrin binding of the RGD finger 1009:1042 These structural features favor integrin binding of the RGD finger. 22601780 4 33 theme second 462:467 arg1 EGF2 481:484 EGF2 481:484 EGF2 481:484 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 4 33 theme second 462:467 arg1 domain 473:478 the second EGF domain 458:478 the second EGF domain (EGF2) 458:485 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 7 34 theme interdomain 755:765 arg1 site 787:790 an interdomain calcium ion binding site 752:790 an interdomain calcium ion binding site 752:790 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 2 35 theme innate 278:283 arg1 immunity 285:292 innate immunity 278:292 innate immunity 278:292 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 6 36 theme protruding 663:672 arg1 finger 695:700 the RGD finger 687:700 dubbed the RGD finger 680:700 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 6 36 theme protruding 663:672 arg1 loop 674:677 a long protruding loop 656:677 a long protruding loop 656:677 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 9 37 theme RGD 1033:1035 arg1 finger 1037:1042 the RGD finger 1029:1042 the RGD finger 1029:1042 These structural features favor integrin binding of the RGD finger. 22601780 1 38 theme Developmental 86:98 arg1 locus-1 117:123 Developmental endothelial cell locus-1 86:123 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 1 38 theme Developmental 86:98 arg1 Del-1 126:130 Del-1 126:130 Del-1 126:130 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 11 39 theme evolutionary 1222:1233 arg1 insertion 1235:1243 an evolutionary insertion 1219:1243 an evolutionary insertion 1219:1243 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 39 theme evolutionary 1222:1233 arg1 finger 1202:1207 this RGD finger 1193:1207 this RGD finger 1193:1207 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 40 theme database 1141:1148 arg1 search 1150:1155 A database search 1139:1155 A database search for EGF domain sequences 1139:1180 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 2 41 theme integrin 319:326 arg1 β 332:332 integrin α(V)β(3) 319:335 integrin α(V)β(3) 319:335 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 4 42 from motif 449:453 arg1 EGF2 481:484 EGF2 481:484 EGF2 481:484 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 4 42 from motif 449:453 arg1 domain 473:478 the second EGF domain 458:478 the second EGF domain (EGF2) 458:485 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 2 43 with interaction 302:312 arg1 β 332:332 integrin α(V)β(3) 319:335 integrin α(V)β(3) 319:335 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 11 44 theme fat 1306:1308 arg1 globule-EGF 1310:1320 its homologue milk fat globule-EGF 8 1287:1322 its homologue milk fat globule-EGF 8 1287:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 45 theme globule-EGF 1310:1320 arg1 Del-1 1277:1281 Del-1 1277:1281 Del-1 1277:1281 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 45 theme globule-EGF 1310:1320 arg1 globule-EGF 1310:1320 its homologue milk fat globule-EGF 8 1287:1322 its homologue milk fat globule-EGF 8 1287:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 45 theme globule-EGF 1310:1320 arg1 domain 1267:1272 the EGF domain 1259:1272 the EGF domain of Del-1 and its homologue milk fat globule-EGF 8 1259:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 0 46 theme Del-1 18:22 arg1 finger 8:13 The RGD finger 0:13 The RGD finger of Del-1 0:22 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 0 46 theme Del-1 18:22 arg1 feature 47:53 a unique structural feature 27:53 a unique structural feature critical for integrin binding 27:83 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 3 47 theme epidermal 355:363 arg1 factor 372:377 3 epidermal growth factor 353:377 3 epidermal growth factor 353:377 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 6 48 theme β 635:635 arg1 turn 637:640 a type II' β turn 624:640 a type II' β turn 624:640 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 3 49 theme -like 384:388 arg1 repeats 390:396 3 epidermal growth factor (EGF)-like repeats 353:396 3 epidermal growth factor (EGF)-like repeats 353:396 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 0 50 theme unique 29:34 arg1 finger 8:13 The RGD finger 0:13 The RGD finger of Del-1 0:22 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 0 50 theme unique 29:34 arg1 feature 47:53 a unique structural feature 27:53 a unique structural feature critical for integrin binding 27:83 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 6 51 theme RGD 691:693 arg1 finger 695:700 the RGD finger 687:700 dubbed the RGD finger 680:700 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 6 51 theme RGD 691:693 arg1 loop 674:677 a long protruding loop 656:677 a long protruding loop 656:677 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 7 52 theme considerable 837:848 arg1 flexibility 850:860 considerable flexibility 837:860 considerable flexibility 837:860 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 2 53 theme α 328:328 arg1 β 332:332 integrin α(V)β(3) 319:335 integrin α(V)β(3) 319:335 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 8 54 theme N-linked 905:912 arg1 glycan 914:919 1 N-linked glycan 903:919 1 N-linked glycan 903:919 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 8 55 from motif 970:974 arg1 side 944:947 the opposite side 931:947 the opposite side of EGF2 from the RGD motif 931:974 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 1 56 theme cell 112:115 arg1 locus-1 117:123 Developmental endothelial cell locus-1 86:123 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 1 56 theme cell 112:115 arg1 Del-1 126:130 Del-1 126:130 Del-1 126:130 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 10 57 theme Mutagenesis 1045:1055 arg1 data 1057:1060 Mutagenesis data 1045:1060 Mutagenesis data 1045:1060 Mutagenesis data confirm the importance of having the RGD motif at the tip of the RGD finger. 22601780 5 58 theme EGF 583:585 arg1 domains 587:593 its 3 EGF domains 577:593 its 3 EGF domains 577:593 We report the crystal structure of its 3 EGF domains. 22601780 6 59 theme dubbed 680:685 arg1 finger 695:700 the RGD finger 687:700 dubbed the RGD finger 680:700 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 6 59 theme dubbed 680:685 arg1 loop 674:677 a long protruding loop 656:677 a long protruding loop 656:677 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 11 60 theme homologue 1291:1299 arg1 globule-EGF 1310:1320 its homologue milk fat globule-EGF 8 1287:1322 its homologue milk fat globule-EGF 8 1287:1322 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 6 61 theme type 626:629 arg1 turn 637:640 a type II' β turn 624:640 a type II' β turn 624:640 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 1 62 theme locus-1 117:123 arg1 glycoprotein 133:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 7 63 theme long 797:800 arg1 linker 802:807 the long linker 793:807 the long linker between EGF1 and EGF2 793:829 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 1 64 theme macrophages 195:205 arg1 cells 173:177 endothelial cells 161:177 endothelial cells 161:177 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 1 64 theme macrophages 195:205 arg1 subset 185:190 a subset 183:190 a subset of macrophages 183:205 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 1 65 gly glycoprotein 133:144 arg1 glycoprotein 133:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein 86:144 Developmental endothelial cell locus-1 (Del-1) glycoprotein is secreted by endothelial cells and a subset of macrophages. 22601780 3 66 dep -like 384:388 arg1 factor 372:377 3 epidermal growth factor 353:377 3 epidermal growth factor 353:377 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 8 67 theme opposite 935:942 arg1 side 944:947 the opposite side 931:947 the opposite side of EGF2 from the RGD motif 931:974 Two unique O-linked glycans and 1 N-linked glycan locate to the opposite side of EGF2 from the RGD motif. 22601780 3 68 theme discoidin-like 404:417 arg1 domains 419:425 2 discoidin-like domains 402:425 2 discoidin-like domains 402:425 Del-1 contains 3 epidermal growth factor (EGF)-like repeats and 2 discoidin-like domains. 22601780 10 69 theme finger 1131:1136 arg1 tip 1116:1118 the tip 1112:1118 the tip of the RGD finger 1112:1136 Mutagenesis data confirm the importance of having the RGD motif at the tip of the RGD finger. 22601780 11 70 theme domain 1165:1170 arg1 sequences 1172:1180 EGF domain sequences 1161:1180 EGF domain sequences 1161:1180 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 6 71 theme loop 674:677 arg1 tip 649:651 the tip 645:651 the tip of a long protruding loop, dubbed the RGD finger 645:700 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 2 72 theme regulatory 222:231 arg1 role 233:236 a regulatory role 220:236 a regulatory role 220:236 Del-1 plays a regulatory role in vascular remodeling and functions in innate immunity through interaction with integrin α(V)β(3). 22601780 7 73 theme rigid 738:742 arg1 rod 744:746 a rigid rod 736:746 a rigid rod 736:746 Whereas EGF2 and EGF3 constitute a rigid rod via an interdomain calcium ion binding site, the long linker between EGF1 and EGF2 lends considerable flexibility to EGF1. 22601780 0 74 theme structural 36:45 arg1 finger 8:13 The RGD finger 0:13 The RGD finger of Del-1 0:22 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 0 74 theme structural 36:45 arg1 feature 47:53 a unique structural feature 27:53 a unique structural feature critical for integrin binding 27:83 The RGD finger of Del-1 is a unique structural feature critical for integrin binding. 22601780 4 75 theme Arg-Gly-Asp 431:441 arg1 motif 449:453 An Arg-Gly-Asp (RGD) motif 428:453 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) 428:485 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 6 76 theme EGF2 613:616 arg1 motif 604:608 The RGD motif 596:608 The RGD motif of EGF2 596:616 The RGD motif of EGF2 forms a type II' β turn at the tip of a long protruding loop, dubbed the RGD finger. 22601780 4 77 theme endothelial 508:518 arg1 cells 520:524 endothelial cells 508:524 endothelial cells 508:524 An Arg-Gly-Asp (RGD) motif in the second EGF domain (EGF2) mediates adhesion by endothelial cells and phagocytes. 22601780 11 78 theme RGD 1198:1200 arg1 insertion 1235:1243 an evolutionary insertion 1219:1243 an evolutionary insertion 1219:1243 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 22601780 11 78 theme RGD 1198:1200 arg1 finger 1202:1207 this RGD finger 1193:1207 this RGD finger 1193:1207 A database search for EGF domain sequences shows that this RGD finger is likely an evolutionary insertion and unique to the EGF domain of Del-1 and its homologue milk fat globule-EGF 8. 8626443 5 0 theme ACET 1120:1123 arg1 processing 1099:1108 processing 1099:1108 processing 1099:1108 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 5 0 theme ACET 1120:1123 arg1 synthesis 1085:1093 synthesis 1085:1093 synthesis 1085:1093 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 1 1 gly unglycosylated 338:351 arg1 proteins 319:326 enzymatically active ACET proteins 293:326 enzymatically active ACET proteins which are unglycosylated or underglycosylated 293:372 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 3 2 theme HeLa 545:548 arg1 cells 550:554 HeLa cells 545:554 HeLa cells 545:554 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 4 3 used used 776:779 arg2 variants 689:696 Several ACET variants 676:696 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites 676:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 0 4 theme enzyme 102:107 arg1 synthesis 45:53 the synthesis 41:53 the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast 41:136 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 4 5 theme cell 872:875 arg1 surface 877:883 the cell surface 868:883 the cell surface 868:883 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 9 6 theme ACE 1441:1443 arg1 proteins 1445:1452 mutant ACE proteins 1434:1452 mutant ACE proteins 1434:1452 Wild type and mutant ACE proteins were cleavage-secreted from yeast and enzymatically active. 8626443 5 7 theme normal 1078:1083 arg1 synthesis 1085:1093 synthesis 1085:1093 synthesis 1085:1093 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 6 8 dep unglycosylated 1206:1219 arg1 inactive 1236:1243 inactive 1236:1243 inactive 1236:1243 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 4 9 gly N-glycosylation 749:763 arg2 sites 765:769 the potential N-glycosylation sites 735:769 the potential N-glycosylation sites 735:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 10 theme cleavage 886:893 arg1 processing 895:904 cleavage processing 886:904 cleavage processing 886:904 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 0 11 theme mammalian 112:120 arg1 cells 122:126 mammalian cells 112:126 mammalian cells 112:126 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 9 12 theme Wild 1420:1423 arg1 type 1425:1428 Wild type 1420:1428 Wild type 1420:1428 Wild type and mutant ACE proteins were cleavage-secreted from yeast and enzymatically active. 8626443 5 13 gly glycosylation 973:985 arg1 first 999:1003 first 999:1003 first 999:1003 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 2 14 from Expression 375:384 arg1 coli 401:404 Escherichia coli 389:404 Escherichia coli of the rabbit ACET cDNA 389:428 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 1 15 theme isozyme 213:219 arg1 crystallization 156:170 crystallization 156:170 crystallization 156:170 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 1 15 theme isozyme 213:219 arg1 studies 187:193 structural studies 176:193 structural studies 176:193 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 4 16 theme ACET 684:687 arg1 variants 689:696 Several ACET variants 676:696 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites 676:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 0 17 from synthesis 45:53 arg1 cells 122:126 mammalian cells 112:126 mammalian cells 112:126 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 0 17 from synthesis 45:53 arg1 yeast 132:136 yeast 132:136 yeast 132:136 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 2 18 theme inactive 481:488 arg1 protein 490:496 an unglycosylated but inactive protein 459:496 an unglycosylated but inactive protein 459:496 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 6 19 contain had 1153:1155 arg1 ACETg3 1139:1144 ACETg3 1139:1144 ACETg3 1139:1144 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 6 19 contain had 1153:1155 arg2 site 1186:1189 only the third glycosylation site 1157:1189 only the third glycosylation site 1157:1189 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 1 20 theme testicular 202:211 arg1 isozyme 213:219 the testicular isozyme 198:219 the testicular isozyme of angiotensin-converting enzyme (ACE,) 198:259 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 0 21 theme glycosylation 10:22 arg1 requirements 24:35 Different glycosylation requirements 0:35 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 0:137 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 6 22 gly glycosylation 1172:1184 arg2 site 1186:1189 only the third glycosylation site 1157:1189 only the third glycosylation site 1157:1189 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 5 23 theme second 1012:1017 arg1 sufficient 1063:1072 sufficient 1063:1072 sufficient 1063:1072 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 5 23 theme second 1012:1017 arg1 site 1019:1022 the second site 1008:1022 the second site 1008:1022 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 4 24 theme sites 765:769 arg1 one 720:722 one 720:722 one 720:722 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 24 theme sites 765:769 arg1 sites 765:769 the potential N-glycosylation sites 735:769 the potential N-glycosylation sites 735:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 2 25 theme ACET 420:423 arg1 cDNA 425:428 the rabbit ACET cDNA 409:428 the rabbit ACET cDNA 409:428 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 0 26 theme Different 0:8 arg1 requirements 24:35 Different glycosylation requirements 0:35 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 0:137 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 6 27 theme glycosylation 1172:1184 arg1 site 1186:1189 only the third glycosylation site 1157:1189 only the third glycosylation site 1157:1189 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 9 28 from yeast 1482:1486 arg1 cleavage-secreted 1459:1475 cleavage-secreted 1459:1475 cleavage-secreted 1459:1475 Wild type and mutant ACE proteins were cleavage-secreted from yeast and enzymatically active. 8626443 3 29 theme potential 591:599 arg1 sites 617:621 all five potential N-glycosylation sites 582:621 all five potential N-glycosylation sites 582:621 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 5 30 dep site 1019:1022 arg1 counted 1028:1034 counted 1028:1034 counted from the NH2 terminus 1028:1056 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 2 31 theme rabbit 413:418 arg1 cDNA 425:428 the rabbit ACET cDNA 409:428 the rabbit ACET cDNA 409:428 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 2 32 theme unglycosylated 462:475 arg1 protein 490:496 an unglycosylated but inactive protein 459:496 an unglycosylated but inactive protein 459:496 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 1 33 theme active 307:312 arg1 proteins 319:326 enzymatically active ACET proteins 293:326 enzymatically active ACET proteins which are unglycosylated or underglycosylated 293:372 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 6 34 from unglycosylated 1206:1219 arg1 contrast 1129:1136 contrast 1129:1136 contrast 1129:1136 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 6 35 from contrast 1129:1136 arg1 unglycosylated 1206:1219 unglycosylated 1206:1219 unglycosylated 1206:1219 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 1 36 theme angiotensin-converting 224:245 arg1 enzyme 247:252 angiotensin-converting enzyme 224:252 angiotensin-converting enzyme (ACE,) 224:259 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 4 37 theme N-glycosylation 749:763 arg1 sites 765:769 the potential N-glycosylation sites 735:769 the potential N-glycosylation sites 735:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 1 38 theme ACET 314:317 arg1 proteins 319:326 enzymatically active ACET proteins 293:326 enzymatically active ACET proteins which are unglycosylated or underglycosylated 293:372 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 3 39 gly unglycosylated 510:523 arg1 ACET 525:528 unglycosylated ACET 510:528 unglycosylated ACET synthesized in HeLa cells 510:554 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 1 40 theme enzyme 247:252 arg1 isozyme 213:219 the testicular isozyme 198:219 the testicular isozyme of angiotensin-converting enzyme (ACE,) 198:259 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 1 41 theme proteins 319:326 arg1 production 279:288 the production 275:288 the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated 275:372 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 6 42 theme third 1166:1170 arg1 site 1186:1189 only the third glycosylation site 1157:1189 only the third glycosylation site 1157:1189 In contrast, ACETg3, which had only the third glycosylation site available, was unglycosylated, enzymatically inactive and rapidly degraded. 8626443 3 43 gly N-glycosylation 601:615 arg2 five 586:589 five 586:589 five 586:589 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 3 43 gly N-glycosylation 601:615 arg2 sites 617:621 all five potential N-glycosylation sites 582:621 all five potential N-glycosylation sites 582:621 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 5 44 theme NH2 1045:1047 arg1 terminus 1049:1056 the NH2 terminus 1041:1056 the NH2 terminus 1041:1056 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 3 45 theme N-glycosylation 601:615 arg1 sites 617:621 all five potential N-glycosylation sites 582:621 all five potential N-glycosylation sites 582:621 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 7 46 theme N-Glycosylated 1267:1280 arg1 ACET 1282:1285 N-Glycosylated ACET 1267:1285 N-Glycosylated ACET 1267:1285 N-Glycosylated ACET could also be produced in yeast. 8626443 2 47 theme protein 490:496 arg1 synthesis 446:454 the synthesis 442:454 the synthesis of an unglycosylated but inactive protein 442:496 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 4 48 theme ACET 839:842 arg1 synthesis 844:852 ACET synthesis 839:852 ACET synthesis 839:852 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 1 49 dep enzyme 247:252 arg1 ACE 255:257 ACE 255:257 ACE 255:257 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 8626443 4 50 theme enzyme 911:916 arg1 activity 918:925 enzyme activity 911:925 enzyme activity 911:925 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 51 contain carrying 698:705 arg2 mutations 707:715 mutations 707:715 mutations 707:715 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 51 contain carrying 698:705 arg1 variants 689:696 Several ACET variants 676:696 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites 676:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 52 theme glycosylation 804:816 arg1 role 796:799 the role 792:799 the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity 792:925 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 53 from sites 830:834 arg1 processing 895:904 cleavage processing 886:904 cleavage processing 886:904 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 53 from sites 830:834 arg1 synthesis 844:852 ACET synthesis 839:852 ACET synthesis 839:852 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 53 from sites 830:834 arg1 transport 855:863 transport 855:863 transport to the cell surface 855:883 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 53 from sites 830:834 arg1 activity 918:925 enzyme activity 911:925 enzyme activity 911:925 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 53 from sites 830:834 arg1 role 796:799 the role 792:799 the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity 792:925 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 3 54 theme unglycosylated 510:523 arg1 ACET 525:528 unglycosylated ACET 510:528 unglycosylated ACET synthesized in HeLa cells 510:554 Similarly, unglycosylated ACET synthesized in HeLa cells, by using a cDNA in which all five potential N-glycosylation sites had been mutated, was inactive and rapidly degraded. 8626443 8 55 theme mutant 1338:1343 arg1 ACETg3 1345:1350 the mutant ACETg3 1334:1350 the mutant ACETg3 1334:1350 Surprisingly, the mutant ACETg3 was synthesized, N-glycosylated, and properly transported in yeast. 8626443 9 56 theme mutant 1434:1439 arg1 proteins 1445:1452 mutant ACE proteins 1434:1452 mutant ACE proteins 1434:1452 Wild type and mutant ACE proteins were cleavage-secreted from yeast and enzymatically active. 8626443 4 57 theme potential 739:747 arg1 sites 765:769 the potential N-glycosylation sites 735:769 the potential N-glycosylation sites 735:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 0 58 theme angiotensin-converting 79:100 arg1 enzyme 102:107 enzymatically active angiotensin-converting enzyme 58:107 enzymatically active angiotensin-converting enzyme 58:107 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 4 59 theme specific 821:828 arg1 sites 830:834 specific sites 821:834 specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity 821:925 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 4 60 theme Several 676:682 arg1 variants 689:696 Several ACET variants 676:696 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites 676:769 Several ACET variants carrying mutations in one or more of the potential N-glycosylation sites were used to examine the role of glycosylation at specific sites on ACET synthesis, transport to the cell surface, cleavage processing, and enzyme activity. 8626443 0 61 theme active 72:77 arg1 enzyme 102:107 enzymatically active angiotensin-converting enzyme 58:107 enzymatically active angiotensin-converting enzyme 58:107 Different glycosylation requirements for the synthesis of enzymatically active angiotensin-converting enzyme in mammalian cells and yeast. 8626443 5 62 theme active 1113:1118 arg1 ACET 1120:1123 active ACET 1113:1123 active ACET 1113:1123 These experiments demonstrated that allowing glycosylation only at the first or the second site, as counted from the NH2 terminus, was sufficient for normal synthesis and processing of active ACET. 8626443 2 63 gly unglycosylated 462:475 arg1 protein 490:496 an unglycosylated but inactive protein 459:496 an unglycosylated but inactive protein 459:496 Expression in Escherichia coli of the rabbit ACET cDNA resulted in the synthesis of an unglycosylated but inactive protein. 8626443 1 64 theme structural 176:185 arg1 studies 187:193 structural studies 176:193 structural studies 176:193 For facilitating crystallization and structural studies of the testicular isozyme of angiotensin-converting enzyme (ACE,), we attempted the production of enzymatically active ACET proteins which are unglycosylated or underglycosylated. 23723439 2 0 attach attached 420:427 arg2 glycans 412:418 O-mannosyl glycans 401:418 O-mannosyl glycans attached to its extracellular α-DG subunit 401:461 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 2 0 attach attached 420:427 arg1 subunit 455:461 its extracellular α-DG subunit 432:461 its extracellular α-DG subunit 432:461 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 9 1 theme HNK-1ST 1748:1754 arg1 actions 1737:1743 opposing actions 1728:1743 opposing actions of HNK-1ST and LARGE 1728:1764 These results illustrate one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE. 23723439 2 2 theme O-mannosyl 401:410 arg1 glycans 412:418 O-mannosyl glycans 401:418 O-mannosyl glycans attached to its extracellular α-DG subunit 401:461 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 8 3 dep in 1513:1514 arg1 vitro 1516:1520 vitro 1516:1520 vitro 1516:1520 Furthermore, using an in vitro enzymatic assay system, we demonstrated that the sulfated α-DG by HNK-1ST is no longer glycosylated by LARGE. 23723439 2 4 theme dystrophy 548:556 arg1 dystroglycanopathy 494:511 dystroglycanopathy 494:511 dystroglycanopathy 494:511 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 2 4 theme dystrophy 548:556 arg1 subclass 516:523 a subclass 514:523 a subclass of congenital muscular dystrophy 514:556 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 4 5 contain had 967:969 arg1 HNK-1ST 869:875 HNK-1ST 869:875 HNK-1ST 869:875 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 5 contain had 967:969 arg2 ability 974:980 an ability to suppress the glycosylation and the function of α-DG 971:1035 an ability to suppress the glycosylation and the function of α-DG 971:1035 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 5 contain had 967:969 arg1 enzymes 923:929 the enzymes 919:929 the enzymes responsible for HNK-1 glycoepitope 919:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 5 contain had 967:969 arg1 sulfotransferase 851:866 human natural killer-1 sulfotransferase 828:866 human natural killer-1 sulfotransferase (HNK-1ST) 828:876 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 5 contain had 967:969 arg1 one 912:914 one 912:914 one 912:914 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 1 6 theme signaling 293:301 arg1 axes 303:306 mechanical and signaling axes 278:306 mechanical and signaling axes in various physiological events 278:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 7 7 theme post-phosphoryl 1279:1293 arg1 site 1308:1311 the single post-phosphoryl modification site 1268:1311 the single post-phosphoryl modification site 1268:1311 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 7 8 theme sulfate 1427:1433 arg1 group 1435:1439 a sulfate group 1425:1439 a sulfate group 1425:1439 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 9 9 theme LARGE 1760:1764 arg1 actions 1737:1743 opposing actions 1728:1743 opposing actions of HNK-1ST and LARGE 1728:1764 These results illustrate one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE. 23723439 8 10 theme in 1513:1514 arg1 system 1538:1543 an in vitro enzymatic assay system 1510:1543 an in vitro enzymatic assay system 1510:1543 Furthermore, using an in vitro enzymatic assay system, we demonstrated that the sulfated α-DG by HNK-1ST is no longer glycosylated by LARGE. 23723439 7 11 theme modification 1295:1306 arg1 site 1308:1311 the single post-phosphoryl modification site 1268:1311 the single post-phosphoryl modification site 1268:1311 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 4 12 gly glycosylation 998:1010 arg1 α-DG 1032:1035 α-DG 1032:1035 α-DG 1032:1035 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 3 13 from modification 714:725 arg1 O-mannose 730:738 O-mannose 730:738 O-mannose 730:738 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 7 14 theme complete 1353:1360 arg1 inhibition 1362:1371 an almost complete inhibition 1343:1371 an almost complete inhibition of the LARGE-dependent modification 1343:1407 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 6 15 theme capable 1220:1226 arg1 residue 1212:1218 only one threonine residue 1193:1218 only one threonine residue capable of being modified by LARGE 1193:1253 We generated an α-DG mutant which has only one threonine residue capable of being modified by LARGE. 23723439 4 16 theme HNK-1 947:951 arg1 glycoepitope 953:964 HNK-1 glycoepitope 947:964 HNK-1 glycoepitope 947:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 6 17 theme α-DG 1171:1174 arg1 mutant 1176:1181 an α-DG mutant 1168:1181 an α-DG mutant which has only one threonine residue capable of being modified by LARGE 1168:1253 We generated an α-DG mutant which has only one threonine residue capable of being modified by LARGE. 23723439 2 18 theme α-DG 450:453 arg1 subunit 455:461 its extracellular α-DG subunit 432:461 its extracellular α-DG subunit 432:461 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 7 19 theme single 1272:1277 arg1 site 1308:1311 the single post-phosphoryl modification site 1268:1311 the single post-phosphoryl modification site 1268:1311 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 6 20 theme threonine 1202:1210 arg1 residue 1212:1218 only one threonine residue 1193:1218 only one threonine residue capable of being modified by LARGE 1193:1253 We generated an α-DG mutant which has only one threonine residue capable of being modified by LARGE. 23723439 3 21 from O-mannose 730:738 arg1 biosynthesis 674:685 the biosynthesis 670:685 the biosynthesis of a phosphodiester-linked modification on O-mannose 670:738 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 1 22 theme extracellular 197:209 arg1 molecules 218:226 extracellular matrix molecules 197:226 extracellular matrix molecules 197:226 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 0 23 theme sulfotransferase-dependent 6:31 arg1 sulfation 33:41 HNK-1 sulfotransferase-dependent sulfation 0:41 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans 0:76 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans occurs in the post-phosphoryl moiety on α-dystroglycan. 23723439 9 24 theme possible 1661:1668 arg1 pathway 1684:1690 one possible glycosylation pathway 1657:1690 one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE 1657:1764 These results illustrate one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE. 23723439 7 25 from moiety 1470:1475 arg1 O-mannose 1480:1488 O-mannose 1480:1488 O-mannose 1480:1488 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 1 26 theme matrix 211:216 arg1 molecules 218:226 extracellular matrix molecules 197:226 extracellular matrix molecules 197:226 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 0 27 theme HNK-1 0:4 arg1 sulfation 33:41 HNK-1 sulfotransferase-dependent sulfation 0:41 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans 0:76 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans occurs in the post-phosphoryl moiety on α-dystroglycan. 23723439 1 28 theme various 311:317 arg1 events 333:338 various physiological events 311:338 various physiological events 311:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 8 29 gly glycosylated 1609:1620 arg1 α-DG 1580:1583 the sulfated α-DG 1567:1583 the sulfated α-DG by HNK-1ST 1567:1594 Furthermore, using an in vitro enzymatic assay system, we demonstrated that the sulfated α-DG by HNK-1ST is no longer glycosylated by LARGE. 23723439 2 30 theme ligand-binding 351:364 arg1 activity 366:373 the ligand-binding activity 347:373 the ligand-binding activity of DG 347:379 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 3 31 from biosynthesis 674:685 arg1 O-mannose 730:738 O-mannose 730:738 O-mannose 730:738 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 7 32 link phosphodiester-linked 1448:1468 arg1 moiety 1470:1475 the phosphodiester-linked moiety 1444:1475 the phosphodiester-linked moiety on O-mannose 1444:1488 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 1 33 theme physiological 319:331 arg1 events 333:338 various physiological events 311:338 various physiological events 311:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 1 34 from axes 303:306 arg1 events 333:338 various physiological events 311:338 various physiological events 311:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 3 35 dep like-acetylglucosaminyltransferase 592:625 arg1 LARGE 628:632 LARGE 628:632 LARGE 628:632 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 7 36 theme modification 1396:1407 arg1 inhibition 1362:1371 an almost complete inhibition 1343:1371 an almost complete inhibition of the LARGE-dependent modification 1343:1407 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 4 37 theme killer-1 842:849 arg1 one 912:914 one 912:914 one 912:914 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 37 theme killer-1 842:849 arg1 enzymes 923:929 the enzymes 919:929 the enzymes responsible for HNK-1 glycoepitope 919:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 37 theme killer-1 842:849 arg1 sulfotransferase 851:866 human natural killer-1 sulfotransferase 828:866 human natural killer-1 sulfotransferase (HNK-1ST) 828:876 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 37 theme killer-1 842:849 arg1 HNK-1ST 869:875 HNK-1ST 869:875 HNK-1ST 869:875 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 2 38 theme aberrant 464:471 arg1 glycosylation 473:485 aberrant glycosylation 464:485 aberrant glycosylation 464:485 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 2 39 theme muscular 539:546 arg1 dystrophy 548:556 congenital muscular dystrophy 528:556 congenital muscular dystrophy 528:556 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 4 40 theme α-DG 1032:1035 arg1 glycosylation 998:1010 the glycosylation 994:1010 the glycosylation 994:1010 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 40 theme α-DG 1032:1035 arg1 function 1020:1027 the function 1016:1027 the function of α-DG 1016:1035 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 0 41 from moiety 108:113 arg1 α-dystroglycan 118:131 α-dystroglycan 118:131 α-dystroglycan 118:131 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans occurs in the post-phosphoryl moiety on α-dystroglycan. 23723439 9 42 theme α-DG 1698:1701 arg1 function 1703:1710 α-DG function 1698:1710 α-DG function 1698:1710 These results illustrate one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE. 23723439 1 43 theme intracellular 235:247 arg1 cytoskeleton 249:260 the intracellular cytoskeleton 231:260 the intracellular cytoskeleton 231:260 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 0 44 theme laminin-binding 54:68 arg1 glycans 70:76 laminin-binding glycans 54:76 laminin-binding glycans 54:76 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans occurs in the post-phosphoryl moiety on α-dystroglycan. 23723439 2 45 theme congenital 528:537 arg1 dystrophy 548:556 congenital muscular dystrophy 528:556 congenital muscular dystrophy 528:556 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 3 46 theme phosphodiester-linked 692:712 arg1 modification 714:725 a phosphodiester-linked modification 690:725 a phosphodiester-linked modification on O-mannose 690:738 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 7 47 theme LARGE-dependent 1380:1394 arg1 modification 1396:1407 the LARGE-dependent modification 1376:1407 the LARGE-dependent modification 1376:1407 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 5 48 gly glycosylation 1095:1107 arg1 α-DG 1112:1115 α-DG 1112:1115 α-DG using deletion and mutation analyses 1112:1152 In this study, we investigated how HNK-1ST regulates the glycosylation of α-DG using deletion and mutation analyses. 23723439 4 49 theme responsible 931:941 arg1 enzymes 923:929 the enzymes 919:929 the enzymes responsible for HNK-1 glycoepitope 919:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 3 50 theme Accumulating 559:570 arg1 evidence 572:579 Accumulating evidence 559:579 Accumulating evidence 559:579 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 3 51 link phosphodiester-linked 692:712 arg1 modification 714:725 a phosphodiester-linked modification 690:725 a phosphodiester-linked modification on O-mannose 690:738 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 8 52 theme sulfated 1571:1578 arg1 α-DG 1580:1583 the sulfated α-DG 1567:1583 the sulfated α-DG by HNK-1ST 1567:1594 Furthermore, using an in vitro enzymatic assay system, we demonstrated that the sulfated α-DG by HNK-1ST is no longer glycosylated by LARGE. 23723439 4 53 theme enzymes 923:929 arg1 one 912:914 one 912:914 one 912:914 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 53 theme enzymes 923:929 arg1 enzymes 923:929 the enzymes 919:929 the enzymes responsible for HNK-1 glycoepitope 919:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 53 theme enzymes 923:929 arg1 sulfotransferase 851:866 human natural killer-1 sulfotransferase 828:866 human natural killer-1 sulfotransferase (HNK-1ST) 828:876 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 9 54 theme glycosylation 1670:1682 arg1 pathway 1684:1690 one possible glycosylation pathway 1657:1690 one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE 1657:1764 These results illustrate one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE. 23723439 8 55 theme enzymatic 1522:1530 arg1 system 1538:1543 an in vitro enzymatic assay system 1510:1543 an in vitro enzymatic assay system 1510:1543 Furthermore, using an in vitro enzymatic assay system, we demonstrated that the sulfated α-DG by HNK-1ST is no longer glycosylated by LARGE. 23723439 3 56 theme ligand-binding 775:788 arg1 activity 790:797 the ligand-binding activity 771:797 the ligand-binding activity 771:797 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 7 57 theme phosphodiester-linked 1448:1468 arg1 moiety 1470:1475 the phosphodiester-linked moiety 1444:1475 the phosphodiester-linked moiety on O-mannose 1444:1488 Focusing on the single post-phosphoryl modification site, we found that HNK-1ST showed an almost complete inhibition of the LARGE-dependent modification and transferred a sulfate group to the phosphodiester-linked moiety on O-mannose. 23723439 5 58 theme α-DG 1112:1115 arg1 glycosylation 1095:1107 the glycosylation 1091:1107 the glycosylation of α-DG using deletion and mutation analyses 1091:1152 In this study, we investigated how HNK-1ST regulates the glycosylation of α-DG using deletion and mutation analyses. 23723439 2 59 theme DG 378:379 arg1 activity 366:373 the ligand-binding activity 347:373 the ligand-binding activity of DG 347:379 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 5 60 dep deletion 1123:1130 arg1 analyses 1145:1152 analyses 1145:1152 analyses 1145:1152 In this study, we investigated how HNK-1ST regulates the glycosylation of α-DG using deletion and mutation analyses. 23723439 1 61 gly glycoprotein 170:181 arg1 Dystroglycan 134:145 Dystroglycan 134:145 Dystroglycan (DG) 134:150 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 1 61 gly glycoprotein 170:181 arg1 glycoprotein 170:181 a cell surface glycoprotein 155:181 a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events 155:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 6 62 contain has 1189:1191 arg1 mutant 1176:1181 an α-DG mutant 1168:1181 an α-DG mutant which has only one threonine residue capable of being modified by LARGE 1168:1253 We generated an α-DG mutant which has only one threonine residue capable of being modified by LARGE. 23723439 6 62 contain has 1189:1191 arg2 residue 1212:1218 only one threonine residue 1193:1218 only one threonine residue capable of being modified by LARGE 1193:1253 We generated an α-DG mutant which has only one threonine residue capable of being modified by LARGE. 23723439 9 63 theme opposing 1728:1735 arg1 actions 1737:1743 opposing actions 1728:1743 opposing actions of HNK-1ST and LARGE 1728:1764 These results illustrate one possible glycosylation pathway where α-DG function is regulated by opposing actions of HNK-1ST and LARGE. 23723439 4 64 theme human 828:832 arg1 one 912:914 one 912:914 one 912:914 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 64 theme human 828:832 arg1 enzymes 923:929 the enzymes 919:929 the enzymes responsible for HNK-1 glycoepitope 919:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 64 theme human 828:832 arg1 sulfotransferase 851:866 human natural killer-1 sulfotransferase 828:866 human natural killer-1 sulfotransferase (HNK-1ST) 828:876 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 64 theme human 828:832 arg1 HNK-1ST 869:875 HNK-1ST 869:875 HNK-1ST 869:875 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 65 theme natural 834:840 arg1 one 912:914 one 912:914 one 912:914 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 65 theme natural 834:840 arg1 enzymes 923:929 the enzymes 919:929 the enzymes responsible for HNK-1 glycoepitope 919:964 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 65 theme natural 834:840 arg1 sulfotransferase 851:866 human natural killer-1 sulfotransferase 828:866 human natural killer-1 sulfotransferase (HNK-1ST) 828:876 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 4 65 theme natural 834:840 arg1 HNK-1ST 869:875 HNK-1ST 869:875 HNK-1ST 869:875 We previously reported that human natural killer-1 sulfotransferase (HNK-1ST), which was originally reported as one of the enzymes responsible for HNK-1 glycoepitope, had an ability to suppress the glycosylation and the function of α-DG. 23723439 0 66 theme post-phosphoryl 92:106 arg1 moiety 108:113 the post-phosphoryl moiety 88:113 the post-phosphoryl moiety on α-dystroglycan 88:131 HNK-1 sulfotransferase-dependent sulfation regulating laminin-binding glycans occurs in the post-phosphoryl moiety on α-dystroglycan. 23723439 3 67 theme modification 714:725 arg1 biosynthesis 674:685 the biosynthesis 670:685 the biosynthesis of a phosphodiester-linked modification on O-mannose 670:738 Accumulating evidence shows that like-acetylglucosaminyltransferase (LARGE), a glycosyltransferase involved in the biosynthesis of a phosphodiester-linked modification on O-mannose, is essential for α-DG to gain the ligand-binding activity. 23723439 1 68 theme cell 157:160 arg1 Dystroglycan 134:145 Dystroglycan 134:145 Dystroglycan (DG) 134:150 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 1 68 theme cell 157:160 arg1 glycoprotein 170:181 a cell surface glycoprotein 155:181 a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events 155:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 2 69 theme extracellular 436:448 arg1 subunit 455:461 its extracellular α-DG subunit 432:461 its extracellular α-DG subunit 432:461 Since the ligand-binding activity of DG strictly depends on O-mannosyl glycans attached to its extracellular α-DG subunit, aberrant glycosylation causes dystroglycanopathy, a subclass of congenital muscular dystrophy. 23723439 8 70 theme assay 1532:1536 arg1 system 1538:1543 an in vitro enzymatic assay system 1510:1543 an in vitro enzymatic assay system 1510:1543 Furthermore, using an in vitro enzymatic assay system, we demonstrated that the sulfated α-DG by HNK-1ST is no longer glycosylated by LARGE. 23723439 1 71 theme surface 162:168 arg1 Dystroglycan 134:145 Dystroglycan 134:145 Dystroglycan (DG) 134:150 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 1 71 theme surface 162:168 arg1 glycoprotein 170:181 a cell surface glycoprotein 155:181 a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events 155:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 23723439 1 72 theme mechanical 278:287 arg1 axes 303:306 mechanical and signaling axes 278:306 mechanical and signaling axes in various physiological events 278:338 Dystroglycan (DG) is a cell surface glycoprotein that connects extracellular matrix molecules to the intracellular cytoskeleton, functioning as mechanical and signaling axes in various physiological events. 7574684 2 0 contain contains 346:353 arg1 domain 339:344 The extracellular domain 321:344 The extracellular domain 321:344 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 0 contain contains 346:353 arg2 acids 365:369 398 amino acids 355:369 398 amino acids 355:369 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 5 1 from bonds 871:875 arg1 region 895:900 the C-terminal region 880:900 the C-terminal region 880:900 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 1 2 theme human 145:149 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 0 3 theme N-glycosylation 73:87 arg1 sites 89:93 N-glycosylation sites 73:93 N-glycosylation sites 73:93 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 5 4 theme N-terminal 783:792 arg1 domain 794:799 the N-terminal domain 779:799 the N-terminal domain 779:799 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 1 5 theme neurotrophin 151:162 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 2 6 theme 33.3 517:520 arg1 % 521:521 % 521:521 % 521:521 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 5 7 theme Cysteine 742:749 arg1 clusters 751:758 Cysteine clusters 1 and 2 742:766 Cysteine clusters 1 and 2 located in the N-terminal domain 742:799 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 5 8 from domain 794:799 arg1 located 768:774 located 768:774 located 768:774 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 3 9 theme disulfide 550:558 arg1 linkages 560:567 Six disulfide linkages 546:567 Six disulfide linkages 546:567 Six disulfide linkages were determined to be Cys1-Cys7, Cys5-Cys14, Cys121-Cys145, Cys123-Cys163, Cys187-Cys235, and Cys271-Cys314, respectively. 7574684 2 10 theme carbohydrate 523:534 arg1 moieties 536:543 33.3% carbohydrate moieties 517:543 33.3% carbohydrate moieties 517:543 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 6 11 theme glycosylation 979:991 arg1 sites 993:997 12 potential N-linked glycosylation sites 957:997 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB 957:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 1 12 theme binding 265:271 arg1 activity 273:280 binding activity 265:280 binding activity for brain-derived neurotrophic factor 265:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 2 13 theme % 521:521 arg1 moieties 536:543 33.3% carbohydrate moieties 517:543 33.3% carbohydrate moieties 517:543 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 4 14 theme sulfhydryl 722:731 arg1 residue 733:739 a free sulfhydryl residue 715:739 a free sulfhydryl residue 715:739 Cys300 was detected as a free sulfhydryl residue. 7574684 4 14 theme sulfhydryl 722:731 arg1 Cys300 692:697 Cys300 692:697 Cys300 692:697 Cys300 was detected as a free sulfhydryl residue. 7574684 0 15 gly N-glycosylation 73:87 arg2 sites 89:93 N-glycosylation sites 73:93 N-glycosylation sites 73:93 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 1 16 theme receptor 164:171 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 0 17 theme ligand 100:105 arg1 binding 107:113 ligand binding 100:113 ligand binding 100:113 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 2 18 contain has 375:377 arg1 domain 339:344 The extracellular domain 321:344 The extracellular domain 321:344 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 18 contain has 375:377 arg2 weight 391:396 a molecular weight 379:396 a molecular weight of 60.6 kDa 379:408 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 19 theme extracellular 479:491 arg1 trkB 503:506 trkB 503:506 trkB 503:506 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 19 theme extracellular 479:491 arg1 domain 493:498 the extracellular domain 475:498 the extracellular domain of trkB 475:506 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 1 20 gly glycoprotein 241:252 arg1 glycoprotein 241:252 a glycoprotein 239:252 a glycoprotein possessing binding activity for brain-derived neurotrophic factor 239:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 20 gly glycoprotein 241:252 arg1 domain 133:138 An extracellular domain 116:138 An extracellular domain of a human neurotrophin receptor trkB 116:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 20 gly glycoprotein 241:252 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 5 21 theme Ig-like 932:938 arg1 domain 943:948 the Ig-like C2 domain 928:948 the Ig-like C2 domain 928:948 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 1 22 theme trkB 173:176 arg1 glycoprotein 241:252 a glycoprotein 239:252 a glycoprotein possessing binding activity for brain-derived neurotrophic factor 239:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 22 theme trkB 173:176 arg1 domain 133:138 An extracellular domain 116:138 An extracellular domain of a human neurotrophin receptor trkB 116:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 22 theme trkB 173:176 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 5 23 theme similar 811:817 arg1 type 819:822 a similar type 809:822 a similar type of disulfide structure 809:845 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 6 24 theme trkB 1033:1036 arg1 trkB 1033:1036 trkB 1033:1036 trkB 1033:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 6 24 theme trkB 1033:1036 arg1 domain 1023:1028 the soluble domain 1011:1028 the soluble domain of trkB 1011:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 2 25 theme extracellular 325:337 arg1 domain 339:344 The extracellular domain 321:344 The extracellular domain 321:344 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 0 26 theme Extracellular 0:12 arg1 trkB 46:49 neurotrophin receptor trkB 24:49 neurotrophin receptor trkB 24:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 0 26 theme Extracellular 0:12 arg1 domain 14:19 Extracellular domain 0:19 Extracellular domain of neurotrophin receptor trkB 0:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 6 27 theme N-linked 970:977 arg1 sites 993:997 12 potential N-linked glycosylation sites 957:997 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB 957:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 5 28 from located 768:774 arg1 domain 794:799 the N-terminal domain 779:799 the N-terminal domain 779:799 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 5 29 theme disulfide 861:869 arg1 bonds 871:875 two other disulfide bonds 851:875 two other disulfide bonds in the C-terminal region 851:900 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 5 30 theme C2 940:941 arg1 domain 943:948 the Ig-like C2 domain 928:948 the Ig-like C2 domain 928:948 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 2 31 theme molecular 381:389 arg1 weight 391:396 a molecular weight 379:396 a molecular weight of 60.6 kDa 379:408 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 0 32 theme neurotrophin 24:35 arg1 trkB 46:49 neurotrophin receptor trkB 24:49 neurotrophin receptor trkB 24:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 6 33 theme potential 960:968 arg1 sites 993:997 12 potential N-linked glycosylation sites 957:997 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB 957:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 2 34 theme laser 423:427 arg1 spectrometry 445:456 laser desorption mass spectrometry 423:456 laser desorption mass spectrometry 423:456 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 35 theme desorption 429:438 arg1 spectrometry 445:456 laser desorption mass spectrometry 423:456 laser desorption mass spectrometry 423:456 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 1 36 theme brain-derived 286:298 arg1 factor 313:318 brain-derived neurotrophic factor 286:318 brain-derived neurotrophic factor 286:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 0 37 dep structure 62:70 arg1 trkB 46:49 neurotrophin receptor trkB 24:49 neurotrophin receptor trkB 24:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 0 37 dep structure 62:70 arg1 domain 14:19 Extracellular domain 0:19 Extracellular domain of neurotrophin receptor trkB 0:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 1 38 theme Chinese 195:201 arg1 cells 217:221 Chinese hamster ovary cells 195:221 Chinese hamster ovary cells 195:221 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 0 39 theme trkB 46:49 arg1 trkB 46:49 neurotrophin receptor trkB 24:49 neurotrophin receptor trkB 24:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 0 39 theme trkB 46:49 arg1 domain 14:19 Extracellular domain 0:19 Extracellular domain of neurotrophin receptor trkB 0:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 1 40 theme neurotrophic 300:311 arg1 factor 313:318 brain-derived neurotrophic factor 286:318 brain-derived neurotrophic factor 286:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 41 theme hamster 203:209 arg1 cells 217:221 Chinese hamster ovary cells 195:221 Chinese hamster ovary cells 195:221 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 0 42 theme receptor 37:44 arg1 trkB 46:49 neurotrophin receptor trkB 24:49 neurotrophin receptor trkB 24:49 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 2 43 theme mass 440:443 arg1 spectrometry 445:456 laser desorption mass spectrometry 423:456 laser desorption mass spectrometry 423:456 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 5 44 theme other 855:859 arg1 bonds 871:875 two other disulfide bonds 851:875 two other disulfide bonds in the C-terminal region 851:900 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 1 45 theme ovary 211:215 arg1 cells 217:221 Chinese hamster ovary cells 195:221 Chinese hamster ovary cells 195:221 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 0 46 theme disulfide 52:60 arg1 structure 62:70 disulfide structure 52:70 disulfide structure 52:70 Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding. 7574684 6 47 gly glycosylated 1061:1072 arg1 sites 1042:1046 10 sites 1039:1046 10 sites 1039:1046 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 5 48 theme located 768:774 arg1 clusters 751:758 Cysteine clusters 1 and 2 742:766 Cysteine clusters 1 and 2 located in the N-terminal domain 742:799 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 6 49 link N-linked 970:977 arg1 sites 993:997 12 potential N-linked glycosylation sites 957:997 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB 957:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 2 50 theme amino 359:363 arg1 acids 365:369 398 amino acids 355:369 398 amino acids 355:369 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 6 51 gly glycosylation 979:991 arg2 12 957:958 12 957:958 12 957:958 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 6 51 gly glycosylation 979:991 arg2 sites 993:997 12 potential N-linked glycosylation sites 957:997 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB 957:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 4 52 theme free 717:720 arg1 residue 733:739 a free sulfhydryl residue 715:739 a free sulfhydryl residue 715:739 Cys300 was detected as a free sulfhydryl residue. 7574684 4 52 theme free 717:720 arg1 Cys300 692:697 Cys300 692:697 Cys300 692:697 Cys300 was detected as a free sulfhydryl residue. 7574684 5 53 contain possess 801:807 arg2 type 819:822 a similar type 809:822 a similar type of disulfide structure 809:845 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 5 53 contain possess 801:807 arg1 clusters 751:758 Cysteine clusters 1 and 2 742:766 Cysteine clusters 1 and 2 located in the N-terminal domain 742:799 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 2 54 contain contains 508:515 arg2 moieties 536:543 33.3% carbohydrate moieties 517:543 33.3% carbohydrate moieties 517:543 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 54 contain contains 508:515 arg1 trkB 503:506 trkB 503:506 trkB 503:506 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 54 contain contains 508:515 arg1 domain 493:498 the extracellular domain 475:498 the extracellular domain of trkB 475:506 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 1 55 theme extracellular 119:131 arg1 glycoprotein 241:252 a glycoprotein 239:252 a glycoprotein possessing binding activity for brain-derived neurotrophic factor 239:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 55 theme extracellular 119:131 arg1 domain 133:138 An extracellular domain 116:138 An extracellular domain of a human neurotrophin receptor trkB 116:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 55 theme extracellular 119:131 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 56 contain possessing 254:263 arg1 glycoprotein 241:252 a glycoprotein 239:252 a glycoprotein possessing binding activity for brain-derived neurotrophic factor 239:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 56 contain possessing 254:263 arg1 domain 133:138 An extracellular domain 116:138 An extracellular domain of a human neurotrophin receptor trkB 116:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 56 contain possessing 254:263 arg1 trkB 173:176 a human neurotrophin receptor trkB 143:176 a human neurotrophin receptor trkB 143:176 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 56 contain possessing 254:263 arg2 activity 273:280 binding activity 265:280 binding activity for brain-derived neurotrophic factor 265:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 1 57 link brain-derived 286:298 arg1 factor 313:318 brain-derived neurotrophic factor 286:318 brain-derived neurotrophic factor 286:318 An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. 7574684 2 58 theme kDa 406:408 arg1 weight 391:396 a molecular weight 379:396 a molecular weight of 60.6 kDa 379:408 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 5 59 theme structure 837:845 arg1 type 819:822 a similar type 809:822 a similar type of disulfide structure 809:845 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 5 60 theme C-terminal 884:893 arg1 region 895:900 the C-terminal region 880:900 the C-terminal region 880:900 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7574684 6 61 theme soluble 1015:1021 arg1 trkB 1033:1036 trkB 1033:1036 trkB 1033:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 6 61 theme soluble 1015:1021 arg1 domain 1023:1028 the soluble domain 1011:1028 the soluble domain of trkB 1011:1036 Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated. 7574684 2 62 theme trkB 503:506 arg1 trkB 503:506 trkB 503:506 trkB 503:506 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 2 62 theme trkB 503:506 arg1 domain 493:498 the extracellular domain 475:498 the extracellular domain of trkB 475:506 The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. 7574684 5 63 theme disulfide 827:835 arg1 structure 837:845 disulfide structure 827:845 disulfide structure 827:845 Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. 7518437 0 0 theme site 98:101 arg1 insertion 103:111 glycosylation site insertion 84:111 glycosylation site insertion 84:111 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 4 1 theme N-glycosylation 561:575 arg1 sidedness 542:550 the sidedness 538:550 the sidedness of their N-glycosylation 538:575 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 1 2 theme membrane 192:199 arg1 proteins 201:208 intrinsic membrane proteins 182:208 intrinsic membrane proteins 182:208 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. 7518437 6 3 theme model 1007:1011 arg1 evaluation 968:977 the first experimental evaluation 945:977 the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins 945:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 4 4 theme transmembrane 646:658 arg1 regulator 672:680 transmembrane conductance regulator 646:680 the cystic fibrosis transmembrane conductance regulator (CFTR) 626:687 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 0 5 theme glycosylation 84:96 arg1 insertion 103:111 glycosylation site insertion 84:111 glycosylation site insertion 84:111 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 6 6 theme CFTR 1016:1019 arg1 model 1007:1011 the original topological model 982:1011 the original topological model of CFTR 982:1019 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 1 7 theme proteins 201:208 arg1 structures 168:177 three-dimensional structures 150:177 three-dimensional structures of intrinsic membrane proteins 150:208 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. 7518437 5 8 theme extracytoplasmic 800:815 arg1 loops 817:821 predicted cytoplasmic and extracytoplasmic loops 774:821 predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane 774:886 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 5 9 theme N-glycosylation 727:741 arg1 NXS/T 764:768 NXS/T 764:768 NXS/T 764:768 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 5 9 theme N-glycosylation 727:741 arg1 sequences 753:761 N-glycosylation consensus sequences 727:761 N-glycosylation consensus sequences (NXS/T) 727:769 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 3 10 theme prokaryotic 459:469 arg1 topology 447:454 the topology 443:454 the topology of prokaryotic but not of eukaryotic membrane proteins 443:509 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 4 11 theme sidedness 542:550 arg1 asymmetry 525:533 The absolute asymmetry 512:533 The absolute asymmetry of the sidedness of their N-glycosylation 512:575 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 5 12 theme consensus 743:751 arg1 NXS/T 764:768 NXS/T 764:768 NXS/T 764:768 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 5 12 theme consensus 743:751 arg1 sequences 753:761 N-glycosylation consensus sequences 727:761 N-glycosylation consensus sequences (NXS/T) 727:769 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 2 13 theme topological 333:343 arg1 arrangement 345:355 their two-dimensional topological arrangement 311:355 their two-dimensional topological arrangement in the lipid bilayer 311:376 However, considerable insight can be gained from their two-dimensional topological arrangement in the lipid bilayer. 7518437 4 14 theme conductance 660:670 arg1 regulator 672:680 transmembrane conductance regulator 646:680 the cystic fibrosis transmembrane conductance regulator (CFTR) 626:687 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 6 15 theme topological 995:1005 arg1 model 1007:1011 the original topological model 982:1011 the original topological model of CFTR 982:1019 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 6 16 theme proteins 1177:1184 arg1 topology 1140:1147 the topology 1136:1147 the topology of other mammalian membrane proteins 1136:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 0 17 gly glycosylation 84:96 arg2 site 98:101 glycosylation site insertion 84:111 glycosylation site insertion 84:111 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 0 17 gly glycosylation 84:96 arg2 insertion 103:111 glycosylation site insertion 84:111 glycosylation site insertion 84:111 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 6 18 theme mammalian 1158:1166 arg1 proteins 1177:1184 other mammalian membrane proteins 1152:1184 other mammalian membrane proteins 1152:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 3 19 theme proteins 502:509 arg1 topology 447:454 the topology 443:454 the topology of prokaryotic but not of eukaryotic membrane proteins 443:509 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 5 20 theme hydrophobic 831:841 arg1 sequences 843:851 hydrophobic sequences 831:851 hydrophobic sequences capable of traversing the membrane 831:886 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 6 21 theme original 986:993 arg1 model 1007:1011 the original topological model 982:1011 the original topological model of CFTR 982:1019 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 5 22 dep in 703:704 arg1 vitro 706:710 vitro 706:710 vitro 706:710 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 6 23 theme first 949:953 arg1 evaluation 968:977 the first experimental evaluation 945:977 the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins 945:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 6 24 theme membrane 1168:1175 arg1 proteins 1177:1184 other mammalian membrane proteins 1152:1184 other mammalian membrane proteins 1152:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 5 25 theme membrane 904:911 arg1 topology 913:920 the membrane topology 900:920 the membrane topology of CFTR 900:928 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 3 26 theme Efficient 379:387 arg1 approaches 407:416 Efficient molecular genetic approaches 379:416 Efficient molecular genetic approaches 379:416 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 5 27 theme in 703:704 arg1 mutagenesis 712:722 in vitro mutagenesis 703:722 in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) 703:769 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 3 28 theme eukaryotic 482:491 arg1 proteins 502:509 eukaryotic membrane proteins 482:509 eukaryotic membrane proteins 482:509 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 2 29 theme lipid 364:368 arg1 bilayer 370:376 the lipid bilayer 360:376 the lipid bilayer 360:376 However, considerable insight can be gained from their two-dimensional topological arrangement in the lipid bilayer. 7518437 3 30 theme membrane 493:500 arg1 proteins 502:509 eukaryotic membrane proteins 482:509 eukaryotic membrane proteins 482:509 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 0 31 theme fibrosis 18:25 arg1 regulator 53:61 cystic fibrosis transmembrane conductance regulator 11:61 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 1 32 theme Technical 114:122 arg1 difficulties 124:135 Technical difficulties 114:135 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins 114:208 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. 7518437 0 33 theme cystic 11:16 arg1 regulator 53:61 cystic fibrosis transmembrane conductance regulator 11:61 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 4 34 theme cystic 630:635 arg1 CFTR 683:686 CFTR 683:686 CFTR 683:686 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 4 34 theme cystic 630:635 arg1 fibrosis 637:644 the cystic fibrosis 626:644 the cystic fibrosis transmembrane conductance regulator (CFTR) 626:687 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 2 35 theme two-dimensional 317:331 arg1 arrangement 345:355 their two-dimensional topological arrangement 311:355 their two-dimensional topological arrangement in the lipid bilayer 311:376 However, considerable insight can be gained from their two-dimensional topological arrangement in the lipid bilayer. 7518437 4 36 theme absolute 516:523 arg1 asymmetry 525:533 The absolute asymmetry 512:533 The absolute asymmetry of the sidedness of their N-glycosylation 512:575 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 6 37 theme topology 1140:1147 arg1 evaluation 1122:1131 the in vivo evaluation 1110:1131 the in vivo evaluation of the topology of other mammalian membrane proteins 1110:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 0 38 theme conductance 41:51 arg1 regulator 53:61 cystic fibrosis transmembrane conductance regulator 11:61 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 5 39 theme cytoplasmic 784:794 arg1 loops 817:821 predicted cytoplasmic and extracytoplasmic loops 774:821 predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane 774:886 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 6 40 theme other 1152:1156 arg1 proteins 1177:1184 other mammalian membrane proteins 1152:1184 other mammalian membrane proteins 1152:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 1 41 theme function 252:259 arg1 understanding 229:241 understanding 229:241 understanding of their function 229:259 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. 7518437 0 42 theme transmembrane 27:39 arg1 regulator 53:61 cystic fibrosis transmembrane conductance regulator 11:61 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 6 43 theme experimental 955:966 arg1 evaluation 968:977 the first experimental evaluation 945:977 the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins 945:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 2 44 theme considerable 271:282 arg1 insight 284:290 considerable insight 271:290 considerable insight 271:290 However, considerable insight can be gained from their two-dimensional topological arrangement in the lipid bilayer. 7518437 5 45 theme predicted 774:782 arg1 loops 817:821 predicted cytoplasmic and extracytoplasmic loops 774:821 predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane 774:886 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 0 46 theme membrane 63:70 arg1 topology 72:79 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 5 47 theme sequences 753:761 arg1 mutagenesis 712:722 in vitro mutagenesis 703:722 in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) 703:769 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 3 48 theme genetic 399:405 arg1 approaches 407:416 Efficient molecular genetic approaches 379:416 Efficient molecular genetic approaches 379:416 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 0 49 theme regulator 53:61 arg1 topology 72:79 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 cystic fibrosis transmembrane conductance regulator membrane topology 11:79 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 6 50 theme hydropathy 1037:1046 arg1 algorithms 1048:1057 hydropathy algorithms 1037:1057 hydropathy algorithms 1037:1057 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 4 51 dep fibrosis 637:644 arg1 regulator 672:680 transmembrane conductance regulator 646:680 the cystic fibrosis transmembrane conductance regulator (CFTR) 626:687 The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). 7518437 5 52 theme capable 853:859 arg1 sequences 843:851 hydrophobic sequences 831:851 hydrophobic sequences capable of traversing the membrane 831:886 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 1 53 theme three-dimensional 150:166 arg1 structures 168:177 three-dimensional structures 150:177 three-dimensional structures of intrinsic membrane proteins 150:208 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. 7518437 2 54 from arrangement 345:355 arg1 bilayer 370:376 the lipid bilayer 360:376 the lipid bilayer 360:376 However, considerable insight can be gained from their two-dimensional topological arrangement in the lipid bilayer. 7518437 3 55 theme molecular 389:397 arg1 approaches 407:416 Efficient molecular genetic approaches 379:416 Efficient molecular genetic approaches 379:416 Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. 7518437 6 56 dep in 1114:1115 arg1 vivo 1117:1120 vivo 1117:1120 vivo 1117:1120 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 6 57 theme in 1114:1115 arg1 evaluation 1122:1131 the in vivo evaluation 1110:1131 the in vivo evaluation of the topology of other mammalian membrane proteins 1110:1184 This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins. 7518437 0 58 theme topology 72:79 arg1 Mapping 0:6 Mapping 0:6 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 0:112 Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. 7518437 5 59 theme CFTR 925:928 arg1 topology 913:920 the membrane topology 900:920 the membrane topology of CFTR 900:928 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 7518437 1 60 theme intrinsic 182:190 arg1 proteins 201:208 intrinsic membrane proteins 182:208 intrinsic membrane proteins 182:208 Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. 7518437 5 61 from Insertion 690:698 arg1 loops 817:821 predicted cytoplasmic and extracytoplasmic loops 774:821 predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane 774:886 Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. 12906826 5 0 theme low 1004:1006 arg1 turnover 1008:1015 low turnover 1004:1015 low turnover 1004:1015 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 4 1 theme additional 784:793 arg1 helix 801:805 an additional short helix 781:805 an additional short helix 781:805 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 5 2 theme reaction 1072:1079 arg1 intermediate 1052:1063 the tetrahedral intermediate 1036:1063 the tetrahedral intermediate of the reaction in the crystal 1036:1094 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 0 3 theme dipeptidyl 80:89 arg1 peptidase-IV 91:102 human dipeptidyl peptidase-IV 74:102 human dipeptidyl peptidase-IV 74:102 Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. 12906826 2 4 from ectodomain 298:307 arg1 pastoris 335:342 Pichia pastoris 328:342 Pichia pastoris 328:342 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 1 5 theme main 157:160 arg1 peptidase 130:138 dipeptidyl peptidase IV 119:141 dipeptidyl peptidase IV (DPP-IV) 119:150 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 5 theme main 157:160 arg1 enzyme 203:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 4 6 theme extra 708:712 arg1 molecule 647:654 the molecule 643:654 the molecule that have not been reported for such structures 643:702 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 6 theme extra 708:712 arg1 motif 725:729 an extra beta sheet motif 705:729 an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif 705:838 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 2 7 theme DPP-IV 318:323 arg1 ectodomain 298:307 the ectodomain 294:307 the ectodomain of human DPP-IV in Pichia pastoris 294:342 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 2 7 theme DPP-IV 318:323 arg1 DPP-IV 318:323 human DPP-IV 312:323 human DPP-IV 312:323 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 3 8 theme beta 499:502 arg1 domains 427:433 two domains 423:433 two domains 423:433 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 3 8 theme beta 499:502 arg1 domain 514:519 a beta propeller domain 497:519 a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif 497:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 5 9 theme Glu 845:847 arg1 motif 849:853 The Glu motif 841:853 The Glu motif 841:853 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 4 10 theme short 795:799 arg1 helix 801:805 an additional short helix 781:805 an additional short helix 781:805 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 11 theme such 688:691 arg1 structures 693:702 such structures 688:702 such structures 688:702 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 1 12 theme peptidase 130:138 arg1 Inhibition 105:114 Inhibition 105:114 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, 105:209 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 4 13 theme dimerization 754:765 arg1 interface 767:775 the dimerization interface 750:775 the dimerization interface 750:775 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 3 14 theme hydrolase 477:485 arg1 fold 487:490 an alpha/beta hydrolase fold 463:490 an alpha/beta hydrolase fold 463:490 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 1 15 theme glucagon-like 162:174 arg1 peptidase 130:138 dipeptidyl peptidase IV 119:141 dipeptidyl peptidase IV (DPP-IV) 119:150 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 15 theme glucagon-like 162:174 arg1 enzyme 203:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 5 16 from intermediate 1052:1063 arg1 crystal 1088:1094 the crystal 1084:1094 the crystal 1084:1094 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 3 17 theme catalytic 440:448 arg1 domain 450:455 the catalytic domain 436:455 the catalytic domain 436:455 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 3 17 theme catalytic 440:448 arg1 domains 427:433 two domains 423:433 two domains 423:433 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 1 18 theme peptide 176:182 arg1 peptidase 130:138 dipeptidyl peptidase IV 119:141 dipeptidyl peptidase IV (DPP-IV) 119:150 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 18 theme peptide 176:182 arg1 enzyme 203:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 5 19 theme tetrahedral 1040:1050 arg1 intermediate 1052:1063 the tetrahedral intermediate 1036:1063 the tetrahedral intermediate of the reaction in the crystal 1036:1094 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 3 20 theme propeller 504:512 arg1 domains 427:433 two domains 423:433 two domains 423:433 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 3 20 theme propeller 504:512 arg1 domain 514:519 a beta propeller domain 497:519 a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif 497:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 0 21 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of proline-specific exopeptidase activity 0:57 Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. 12906826 1 22 theme type 250:253 arg1 diabetes 258:265 type II diabetes 250:265 type II diabetes 250:265 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 3 23 with domain 450:455 arg1 repeat 536:541 an 8-fold repeat 526:541 an 8-fold repeat of a four-strand beta sheet motif 526:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 3 23 with domain 450:455 arg1 fold 487:490 an alpha/beta hydrolase fold 463:490 an alpha/beta hydrolase fold 463:490 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 4 24 theme propeller 587:595 arg1 domain 597:602 The beta propeller domain 578:602 The beta propeller domain 578:602 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 0 25 theme proline-specific 20:35 arg1 activity 50:57 proline-specific exopeptidase activity 20:57 proline-specific exopeptidase activity 20:57 Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. 12906826 5 26 theme diprotin 975:982 arg1 A 984:984 diprotin A 975:984 diprotin A 975:984 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 5 26 theme diprotin 975:982 arg1 substrate 989:997 a substrate 987:997 a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal 987:1094 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 2 27 theme A 386:386 arg1 resolution 388:397 2.1 A resolution 382:397 2.1 A resolution 382:397 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 3 28 with domain 514:519 arg1 repeat 536:541 an 8-fold repeat 526:541 an 8-fold repeat of a four-strand beta sheet motif 526:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 3 28 with domain 514:519 arg1 fold 487:490 an alpha/beta hydrolase fold 463:490 an alpha/beta hydrolase fold 463:490 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 1 29 theme diabetes 258:265 arg1 treatment 237:245 the treatment 233:245 the treatment of type II diabetes 233:265 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 4 30 theme sequence 825:832 arg1 motif 834:838 a double Glu sequence motif 812:838 a double Glu sequence motif 812:838 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 5 31 theme complex 952:958 arg1 structure 960:968 the complex structure 948:968 the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal 948:1094 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 1 32 theme dipeptidyl 119:128 arg1 peptidase 130:138 dipeptidyl peptidase IV 119:141 dipeptidyl peptidase IV (DPP-IV) 119:150 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 32 theme dipeptidyl 119:128 arg1 DPP-IV 144:149 DPP-IV 144:149 DPP-IV 144:149 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 32 theme dipeptidyl 119:128 arg1 enzyme 203:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 0 33 theme activity 50:57 arg1 basis 11:15 Structural basis 0:15 Structural basis of proline-specific exopeptidase activity 0:57 Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. 12906826 3 34 theme 8-fold 529:534 arg1 repeat 536:541 an 8-fold repeat 526:541 an 8-fold repeat of a four-strand beta sheet motif 526:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 5 35 with structure 960:968 arg1 A 984:984 diprotin A 975:984 diprotin A 975:984 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 5 35 with structure 960:968 arg1 substrate 989:997 a substrate 987:997 a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal 987:1094 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 1 36 theme -degrading 192:201 arg1 peptidase 130:138 dipeptidyl peptidase IV 119:141 dipeptidyl peptidase IV (DPP-IV) 119:150 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 36 theme -degrading 192:201 arg1 enzyme 203:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 0 37 theme exopeptidase 37:48 arg1 activity 50:57 proline-specific exopeptidase activity 20:57 proline-specific exopeptidase activity 20:57 Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. 12906826 3 38 theme sheet 565:569 arg1 motif 571:575 a four-strand beta sheet motif 546:575 a four-strand beta sheet motif 546:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 3 39 theme motif 571:575 arg1 repeat 536:541 an 8-fold repeat 526:541 an 8-fold repeat of a four-strand beta sheet motif 526:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 5 40 theme substrates 921:930 arg1 terminus 905:912 the N terminus 899:912 the N terminus of the substrates 899:930 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 4 41 theme interface 767:775 arg1 part 742:745 part 742:745 part of the dimerization interface and an additional short helix 742:805 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 2 42 theme human 312:316 arg1 DPP-IV 318:323 human DPP-IV 312:323 human DPP-IV 312:323 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 2 43 theme X-ray 363:367 arg1 structure 369:377 the X-ray structure 359:377 the X-ray structure 359:377 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 4 44 theme helix 801:805 arg1 part 742:745 part 742:745 part of the dimerization interface and an additional short helix 742:805 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 45 theme Glu 821:823 arg1 motif 834:838 a double Glu sequence motif 812:838 a double Glu sequence motif 812:838 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 46 theme beta 714:717 arg1 molecule 647:654 the molecule 643:654 the molecule that have not been reported for such structures 643:702 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 46 theme beta 714:717 arg1 motif 725:729 an extra beta sheet motif 705:729 an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif 705:838 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 3 47 theme four-strand 548:558 arg1 motif 571:575 a four-strand beta sheet motif 546:575 a four-strand beta sheet motif 546:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 5 48 with substrate 989:997 arg1 turnover 1008:1015 low turnover 1004:1015 low turnover 1004:1015 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 4 49 theme double 814:819 arg1 motif 834:838 a double Glu sequence motif 812:838 a double Glu sequence motif 812:838 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 3 50 theme beta 560:563 arg1 motif 571:575 a four-strand beta sheet motif 546:575 a four-strand beta sheet motif 546:575 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 12906826 2 51 theme Pichia 328:333 arg1 pastoris 335:342 Pichia pastoris 328:342 Pichia pastoris 328:342 We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. 12906826 0 52 theme human 74:78 arg1 peptidase-IV 91:102 human dipeptidyl peptidase-IV 74:102 human dipeptidyl peptidase-IV 74:102 Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. 12906826 5 53 theme binding 882:888 arg1 site 890:893 a binding site 880:893 a binding site for the N terminus of the substrates 880:930 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 5 54 theme N 903:903 arg1 terminus 905:912 the N terminus 899:912 the N terminus of the substrates 899:930 The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. 12906826 1 55 theme GLP1 187:190 arg1 peptidase 130:138 dipeptidyl peptidase IV 119:141 dipeptidyl peptidase IV (DPP-IV) 119:150 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 1 55 theme GLP1 187:190 arg1 enzyme 203:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 the main glucagon-like peptide 1 (GLP1)-degrading enzyme 153:208 Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. 12906826 4 56 theme important 620:628 arg1 functions 630:638 two important functions 616:638 two important functions 616:638 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 57 theme beta 582:585 arg1 domain 597:602 The beta propeller domain 578:602 The beta propeller domain 578:602 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 58 theme sheet 719:723 arg1 molecule 647:654 the molecule 643:654 the molecule that have not been reported for such structures 643:702 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 4 58 theme sheet 719:723 arg1 motif 725:729 an extra beta sheet motif 705:729 an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif 705:838 The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. 12906826 3 59 theme alpha/beta 466:475 arg1 fold 487:490 an alpha/beta hydrolase fold 463:490 an alpha/beta hydrolase fold 463:490 The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. 17139081 5 0 theme Cerezyme 717:724 arg1 structure 678:686 The structure 674:686 The structure of intact fully glycosylated Cerezyme 674:724 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 5 0 theme Cerezyme 717:724 arg1 identical 739:747 identical 739:747 identical 739:747 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 7 1 from family 1097:1102 arg1 enzyme 1059:1064 a bacterial enzyme 1047:1064 a bacterial enzyme from a closely related protein family 1047:1102 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 7 1 from family 1097:1102 arg1 xylanase 1037:1044 xylanase 1037:1044 xylanase 1037:1044 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 4 2 theme Cerezyme 532:539 arg1 structure 508:516 The crystal structure 496:516 The crystal structure of unmodified Cerezyme 496:539 The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. 17139081 0 3 theme defective 86:94 arg1 enzyme 96:101 the defective enzyme 82:101 the defective enzyme in Gaucher disease 82:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 0 3 theme defective 86:94 arg1 acid-beta-glucosidase 59:79 acid-beta-glucosidase 59:79 acid-beta-glucosidase 59:79 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 7 4 theme related 1081:1087 arg1 family 1097:1102 a closely related protein family 1071:1102 a closely related protein family 1071:1102 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 7 5 theme protein 1089:1095 arg1 family 1097:1102 a closely related protein family 1071:1102 a closely related protein family 1071:1102 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 4 6 theme unmodified 521:530 arg1 Cerezyme 532:539 unmodified Cerezyme 521:539 unmodified Cerezyme 521:539 The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. 17139081 0 7 theme Gaucher 106:112 arg1 disease 114:120 Gaucher disease 106:120 Gaucher disease 106:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 6 8 theme active 845:850 arg1 site 852:855 the active site 841:855 the active site 841:855 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 2 9 theme enzyme 233:238 arg1 Cerezyme 241:248 Cerezyme 241:248 Cerezyme 241:248 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 2 9 theme enzyme 233:238 arg1 form 220:223 A recombinant form 206:223 A recombinant form of this enzyme 206:238 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 5 10 gly glycosylated 704:715 arg1 Cerezyme 717:724 intact fully glycosylated Cerezyme 691:724 intact fully glycosylated Cerezyme 691:724 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 7 11 with Comparison 971:980 arg1 that 1029:1032 that 1029:1032 that 1029:1032 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 3 12 theme partial 357:363 arg1 deglycosylation 365:379 its partial deglycosylation 353:379 its partial deglycosylation 353:379 Crystals of Cerezyme after its partial deglycosylation were obtained earlier and the structure was solved to 2.0 A resolution [Dvir et al. (2003), EMBO Rep. 4, 704-709]. 17139081 2 13 theme disease 276:282 arg1 patients 284:291 Gaucher disease patients 268:291 Gaucher disease patients 268:291 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 6 14 theme additional 927:936 arg1 variability 938:948 additional variability 927:948 additional variability in their structures 927:968 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 3 15 theme Cerezyme 338:345 arg1 Crystals 326:333 Crystals 326:333 Crystals of Cerezyme after its partial deglycosylation 326:379 Crystals of Cerezyme after its partial deglycosylation were obtained earlier and the structure was solved to 2.0 A resolution [Dvir et al. (2003), EMBO Rep. 4, 704-709]. 17139081 6 16 from variability 938:948 arg1 structures 959:968 their structures 953:968 their structures 953:968 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 1 17 from mutations 152:160 arg1 gene 169:172 the gene 165:172 the gene encoding acid-beta-glucosidase 165:203 Gaucher disease is caused by mutations in the gene encoding acid-beta-glucosidase. 17139081 2 18 theme Gaucher 268:274 arg1 patients 284:291 Gaucher disease patients 268:291 Gaucher disease patients 268:291 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 4 19 theme residues 597:604 arg1 number 581:586 a substantial number 567:586 a substantial number of sugar residues bound to three asparagines via N-glycosylation 567:651 The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. 17139081 5 20 theme intact 691:696 arg1 Cerezyme 717:724 intact fully glycosylated Cerezyme 691:724 intact fully glycosylated Cerezyme 691:724 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 2 21 used used 254:257 arg2 form 220:223 A recombinant form 206:223 A recombinant form of this enzyme 206:238 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 2 21 used used 254:257 arg2 Cerezyme 241:248 Cerezyme 241:248 Cerezyme 241:248 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 4 22 theme sugar 591:595 arg1 residues 597:604 sugar residues 591:604 sugar residues 591:604 The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. 17139081 0 23 theme Structural 0:9 arg1 comparison 11:20 Structural comparison 0:20 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease 0:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 4 24 theme crystal 500:506 arg1 structure 508:516 The crystal structure 496:516 The crystal structure of unmodified Cerezyme 496:539 The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. 17139081 3 25 dep EMBO 473:476 arg1 704-709 486:492 704-709 486:492 704-709 486:492 Crystals of Cerezyme after its partial deglycosylation were obtained earlier and the structure was solved to 2.0 A resolution [Dvir et al. (2003), EMBO Rep. 4, 704-709]. 17139081 4 26 theme substantial 569:579 arg1 number 581:586 a substantial number 567:586 a substantial number of sugar residues bound to three asparagines via N-glycosylation 567:651 The crystal structure of unmodified Cerezyme is now reported, in which a substantial number of sugar residues bound to three asparagines via N-glycosylation could be visualized. 17139081 2 27 theme enzyme-replacement 297:314 arg1 therapy 316:322 enzyme-replacement therapy 297:322 enzyme-replacement therapy 297:322 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 0 28 from enzyme 96:101 arg1 disease 114:120 Gaucher disease 106:120 Gaucher disease 106:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 0 29 theme forms 50:54 arg1 comparison 11:20 Structural comparison 0:20 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease 0:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 6 30 theme alternative 892:902 arg1 conformations 904:916 alternative conformations 892:916 alternative conformations 892:916 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 7 31 theme active-site 1153:1163 arg1 residues 1165:1172 the active-site residues 1149:1172 the active-site residues of the two enzymes 1149:1191 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 7 31 theme active-site 1153:1163 arg1 enzymes 1185:1191 the two enzymes 1177:1191 the two enzymes 1177:1191 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 6 32 located observed 880:887 arg1 conformations 904:916 alternative conformations 892:916 alternative conformations 892:916 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 6 32 located observed 880:887 arg2 site 852:855 the active site 841:855 the active site 841:855 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 5 33 gly deglycosylated 774:787 arg1 enzyme 789:794 the partially deglycosylated enzyme 760:794 the partially deglycosylated enzyme 760:794 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 0 34 theme glycosylated 37:48 arg1 forms 50:54 differently glycosylated forms 25:54 differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease 25:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 3 35 theme [Dvir 452:456 arg1 resolution 441:450 A resolution 439:450 2.0 A resolution [Dvir et al. (2003) 435:470 Crystals of Cerezyme after its partial deglycosylation were obtained earlier and the structure was solved to 2.0 A resolution [Dvir et al. (2003), EMBO Rep. 4, 704-709]. 17139081 0 36 theme acid-beta-glucosidase 59:79 arg1 forms 50:54 differently glycosylated forms 25:54 differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease 25:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 6 37 from entrance 829:836 arg1 loops 816:820 the three loops 806:820 the three loops at the entrance to the active site, which were previously observed in alternative conformations, 806:917 However, the three loops at the entrance to the active site, which were previously observed in alternative conformations, display additional variability in their structures. 17139081 3 38 dep 2.0 435:437 arg1 resolution 441:450 A resolution 439:450 2.0 A resolution [Dvir et al. (2003) 435:470 Crystals of Cerezyme after its partial deglycosylation were obtained earlier and the structure was solved to 2.0 A resolution [Dvir et al. (2003), EMBO Rep. 4, 704-709]. 17139081 1 39 theme Gaucher 123:129 arg1 disease 131:137 Gaucher disease 123:137 Gaucher disease 123:137 Gaucher disease is caused by mutations in the gene encoding acid-beta-glucosidase. 17139081 7 40 theme acid-beta-glucosidase 1002:1022 arg1 structure 989:997 the structure 985:997 the structure of acid-beta-glucosidase 985:1022 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 0 41 gly glycosylated 37:48 arg1 forms 50:54 differently glycosylated forms 25:54 differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease 25:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 0 41 gly glycosylated 37:48 arg1 enzyme 96:101 the defective enzyme 82:101 the defective enzyme in Gaucher disease 82:120 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 0 41 gly glycosylated 37:48 arg1 acid-beta-glucosidase 59:79 acid-beta-glucosidase 59:79 acid-beta-glucosidase 59:79 Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. 17139081 7 42 theme bacterial 1049:1057 arg1 enzyme 1059:1064 a bacterial enzyme 1047:1064 a bacterial enzyme from a closely related protein family 1047:1102 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 7 42 theme bacterial 1049:1057 arg1 xylanase 1037:1044 xylanase 1037:1044 xylanase 1037:1044 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 7 43 theme structure 989:997 arg1 Comparison 971:980 Comparison 971:980 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, 971:1103 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 2 44 theme recombinant 208:218 arg1 Cerezyme 241:248 Cerezyme 241:248 Cerezyme 241:248 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 2 44 theme recombinant 208:218 arg1 form 220:223 A recombinant form 206:223 A recombinant form of this enzyme 206:238 A recombinant form of this enzyme, Cerezyme, is used to treat Gaucher disease patients by ;enzyme-replacement therapy'. 17139081 7 45 theme enzymes 1185:1191 arg1 residues 1165:1172 the active-site residues 1149:1172 the active-site residues of the two enzymes 1149:1191 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 7 45 theme enzymes 1185:1191 arg1 enzymes 1185:1191 the two enzymes 1177:1191 the two enzymes 1177:1191 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 17139081 5 46 theme glycosylated 704:715 arg1 Cerezyme 717:724 intact fully glycosylated Cerezyme 691:724 intact fully glycosylated Cerezyme 691:724 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 5 47 theme deglycosylated 774:787 arg1 enzyme 789:794 the partially deglycosylated enzyme 760:794 the partially deglycosylated enzyme 760:794 The structure of intact fully glycosylated Cerezyme is virtually identical to that of the partially deglycosylated enzyme. 17139081 7 48 theme close 1120:1124 arg1 correspondence 1126:1139 a close correspondence 1118:1139 a close correspondence between the active-site residues of the two enzymes 1118:1191 Comparison of the structure of acid-beta-glucosidase with that of xylanase, a bacterial enzyme from a closely related protein family, demonstrates a close correspondence between the active-site residues of the two enzymes. 16002699 6 0 theme N110 876:879 arg1 sites 895:899 the N78 and N110 glycosylation sites 864:899 sites 895:899 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 7 1 theme N78A/N110A/C122A 954:969 arg1 CTLA-4 985:990 the N78A/N110A/C122A triple mutant CTLA-4 950:990 the N78A/N110A/C122A triple mutant CTLA-4 950:990 Despite being monomeric, the N78A/N110A/C122A triple mutant CTLA-4 localized in the immunological synapse and inhibited T cell activation. 16002699 1 2 theme inhibitory 163:172 arg1 CTLA-4 118:123 CTLA-4 118:123 CTLA-4 118:123 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 1 2 theme inhibitory 163:172 arg1 receptor 174:181 an activation-induced, homodimeric inhibitory receptor 128:181 an activation-induced, homodimeric inhibitory receptor in T cells 128:192 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 6 3 gly glycosylation 881:893 arg2 sites 895:899 the N78 and N110 glycosylation sites 864:899 sites 895:899 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 5 4 theme immunological 714:726 arg1 synapse 728:734 the immunological synapse 710:734 the immunological synapse 710:734 Moreover, C122A CTLA-4 partitioned within lipid rafts, colocalized with the TCR in the immunological synapse, and inhibited T cell activation. 16002699 4 5 dep 35 573:574 arg1 to 570:571 to 570:571 to 570:571 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 3 6 from dimerization 513:524 arg1 solution 529:536 solution 529:536 solution 529:536 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 0 7 theme cell 99:102 arg1 inactivation 104:115 T cell inactivation 97:115 T cell inactivation 97:115 Hierarchical regulation of CTLA-4 dimer-based lattice formation and its biological relevance for T cell inactivation. 16002699 6 8 theme N78 868:870 arg1 mutation 852:859 further mutation 844:859 further mutation of the N78 and N110 glycosylation sites 844:899 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 1 9 from receptor 174:181 arg1 cells 188:192 T cells 186:192 T cells 186:192 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 5 10 from TCR 703:705 arg1 synapse 728:734 the immunological synapse 710:734 the immunological synapse 710:734 Moreover, C122A CTLA-4 partitioned within lipid rafts, colocalized with the TCR in the immunological synapse, and inhibited T cell activation. 16002699 0 11 theme T 97:97 arg1 inactivation 104:115 T cell inactivation 97:115 T cell inactivation 97:115 Hierarchical regulation of CTLA-4 dimer-based lattice formation and its biological relevance for T cell inactivation. 16002699 9 12 theme CTLA-4 1318:1323 arg1 lattices 1325:1332 dimer-dependent CTLA-4 lattices 1302:1332 dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation 1302:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 1 13 dep activation-induced 131:148 arg1 homodimeric 151:161 homodimeric 151:161 homodimeric 151:161 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 9 14 theme lattices 1325:1332 arg1 formation 1289:1297 the formation 1285:1297 the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation 1285:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 8 15 theme molecules 1139:1147 arg1 dimerization 1110:1121 B7-induced dimerization 1099:1121 B7-induced dimerization of these mutant molecules 1099:1147 Such functionality correlated with B7-induced dimerization of these mutant molecules. 16002699 2 16 theme cell 286:289 arg1 surface 291:297 the cell surface 282:297 the cell surface 282:297 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 4 17 from lymphocytes 614:624 arg1 dimerized 593:601 dimerized 593:601 dimerized 593:601 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 3 18 theme C122A 424:428 arg1 mutation 436:443 a C122A point mutation 422:443 a C122A point mutation 422:443 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 9 19 theme hierarchical 1193:1204 arg1 regulation 1206:1215 hierarchical regulation 1193:1215 hierarchical regulation of CTLA-4 oligomerization 1193:1241 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 5 20 theme cell 753:756 arg1 activation 758:767 T cell activation 751:767 T cell activation 751:767 Moreover, C122A CTLA-4 partitioned within lipid rafts, colocalized with the TCR in the immunological synapse, and inhibited T cell activation. 16002699 1 21 theme T 186:186 arg1 cells 188:192 T cells 186:192 T cells 186:192 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 5 22 theme T 751:751 arg1 activation 758:767 T cell activation 751:767 T cell activation 751:767 Moreover, C122A CTLA-4 partitioned within lipid rafts, colocalized with the TCR in the immunological synapse, and inhibited T cell activation. 16002699 6 23 theme further 844:850 arg1 mutation 852:859 further mutation 844:859 further mutation of the N78 and N110 glycosylation sites 844:899 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 4 24 theme human 606:610 arg1 lymphocytes 614:624 human T lymphocytes 606:624 human T lymphocytes 606:624 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 0 25 theme Hierarchical 0:11 arg1 regulation 13:22 Hierarchical regulation 0:22 Hierarchical regulation of CTLA-4 0:32 Hierarchical regulation of CTLA-4 dimer-based lattice formation and its biological relevance for T cell inactivation. 16002699 6 26 theme C122-independent 770:785 arg1 dimerization 787:798 C122-independent dimerization 770:798 C122-independent dimerization of CTLA-4 770:808 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 9 27 theme regulation 1206:1215 arg1 model 1184:1188 a model 1182:1188 a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation 1182:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 9 28 theme B7 1252:1253 arg1 binding 1255:1261 B7 binding 1252:1261 B7 binding 1252:1261 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 3 29 theme biological 358:367 arg1 relevance 369:377 the biological relevance 354:377 the biological relevance of these CTLA-4-B7 lattices 354:405 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 2 30 dep molecules 335:343 arg1 CD86 329:332 CD86 329:332 CD86 329:332 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 2 30 dep molecules 335:343 arg1 CD80 323:326 CD80 323:326 CD80 323:326 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 4 31 from dimerized 593:601 arg1 lymphocytes 614:624 human T lymphocytes 606:624 human T lymphocytes 606:624 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 3 32 theme human 448:452 arg1 CTLA-4 454:459 human CTLA-4 448:459 human CTLA-4 448:459 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 7 33 theme mutant 978:983 arg1 CTLA-4 985:990 the N78A/N110A/C122A triple mutant CTLA-4 950:990 the N78A/N110A/C122A triple mutant CTLA-4 950:990 Despite being monomeric, the N78A/N110A/C122A triple mutant CTLA-4 localized in the immunological synapse and inhibited T cell activation. 16002699 7 34 theme T 1045:1045 arg1 activation 1052:1061 T cell activation 1045:1061 T cell activation 1045:1061 Despite being monomeric, the N78A/N110A/C122A triple mutant CTLA-4 localized in the immunological synapse and inhibited T cell activation. 16002699 8 35 theme Such 1064:1067 arg1 functionality 1069:1081 Such functionality 1064:1081 Such functionality 1064:1081 Such functionality correlated with B7-induced dimerization of these mutant molecules. 16002699 9 36 theme CTLA-4 1220:1225 arg1 oligomerization 1227:1241 CTLA-4 oligomerization 1220:1241 CTLA-4 oligomerization 1220:1241 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 4 37 theme C122A 580:584 arg1 CTLA-4 586:591 C122A CTLA-4 580:591 C122A CTLA-4 580:591 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 0 38 theme lattice 46:52 arg1 formation 54:62 lattice formation 46:62 lattice formation 46:62 Hierarchical regulation of CTLA-4 dimer-based lattice formation and its biological relevance for T cell inactivation. 16002699 5 39 theme lipid 669:673 arg1 rafts 675:679 lipid rafts 669:679 lipid rafts 669:679 Moreover, C122A CTLA-4 partitioned within lipid rafts, colocalized with the TCR in the immunological synapse, and inhibited T cell activation. 16002699 9 40 theme oligomerization 1227:1241 arg1 regulation 1206:1215 hierarchical regulation 1193:1215 hierarchical regulation of CTLA-4 oligomerization 1193:1241 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 2 41 from arrays 272:277 arg1 surface 291:297 the cell surface 282:297 the cell surface 282:297 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 4 42 theme T 612:612 arg1 lymphocytes 614:624 human T lymphocytes 606:624 human T lymphocytes 606:624 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 9 43 theme B7-dependent 1371:1382 arg1 inactivation 1391:1402 B7-dependent T cell inactivation 1371:1402 B7-dependent T cell inactivation 1371:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 7 44 theme cell 1047:1050 arg1 activation 1052:1061 T cell activation 1045:1061 T cell activation 1045:1061 Despite being monomeric, the N78A/N110A/C122A triple mutant CTLA-4 localized in the immunological synapse and inhibited T cell activation. 16002699 4 45 dep found 556:560 arg1 dimerized 593:601 dimerized 593:601 dimerized 593:601 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 0 46 theme CTLA-4 27:32 arg1 regulation 13:22 Hierarchical regulation 0:22 Hierarchical regulation of CTLA-4 0:32 Hierarchical regulation of CTLA-4 dimer-based lattice formation and its biological relevance for T cell inactivation. 16002699 3 47 theme CTLA-4-B7 388:396 arg1 lattices 398:405 these CTLA-4-B7 lattices 382:405 these CTLA-4-B7 lattices 382:405 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 9 48 theme T 1384:1384 arg1 inactivation 1391:1402 B7-dependent T cell inactivation 1371:1402 B7-dependent T cell inactivation 1371:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 7 49 theme immunological 1009:1021 arg1 synapse 1023:1029 the immunological synapse 1005:1029 the immunological synapse 1005:1029 Despite being monomeric, the N78A/N110A/C122A triple mutant CTLA-4 localized in the immunological synapse and inhibited T cell activation. 16002699 3 50 theme lattices 398:405 arg1 relevance 369:377 the biological relevance 354:377 the biological relevance of these CTLA-4-B7 lattices 354:405 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 9 51 theme cell 1386:1389 arg1 inactivation 1391:1402 B7-dependent T cell inactivation 1371:1402 B7-dependent T cell inactivation 1371:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 8 52 theme B7-induced 1099:1108 arg1 dimerization 1110:1121 B7-induced dimerization 1099:1121 B7-induced dimerization of these mutant molecules 1099:1147 Such functionality correlated with B7-induced dimerization of these mutant molecules. 16002699 6 53 theme CTLA-4 803:808 arg1 dimerization 787:798 C122-independent dimerization 770:798 C122-independent dimerization of CTLA-4 770:808 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 6 54 theme sites 895:899 arg1 mutation 852:859 further mutation 844:859 further mutation of the N78 and N110 glycosylation sites 844:899 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 16002699 5 55 theme C122A 637:641 arg1 CTLA-4 643:648 C122A CTLA-4 637:648 C122A CTLA-4 637:648 Moreover, C122A CTLA-4 partitioned within lipid rafts, colocalized with the TCR in the immunological synapse, and inhibited T cell activation. 16002699 4 56 dep dimerized 593:601 arg1 % 575:575 up to 35% 567:575 up to 35% of C122A CTLA-4 567:591 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 4 56 dep dimerized 593:601 arg1 CTLA-4 586:591 C122A CTLA-4 580:591 C122A CTLA-4 580:591 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 9 57 theme dimer-dependent 1302:1316 arg1 lattices 1325:1332 dimer-dependent CTLA-4 lattices 1302:1332 dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation 1302:1402 Based on these data, we propose a model of hierarchical regulation of CTLA-4 oligomerization by which B7 binding ultimately determines the formation of dimer-dependent CTLA-4 lattices that may be necessary for triggering B7-dependent T cell inactivation. 16002699 2 58 theme crystallographic 202:217 arg1 reports 219:225 Recent crystallographic reports 195:225 Recent crystallographic reports 195:225 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 7 59 theme triple 971:976 arg1 CTLA-4 985:990 the N78A/N110A/C122A triple mutant CTLA-4 950:990 the N78A/N110A/C122A triple mutant CTLA-4 950:990 Despite being monomeric, the N78A/N110A/C122A triple mutant CTLA-4 localized in the immunological synapse and inhibited T cell activation. 16002699 1 60 theme activation-induced 131:148 arg1 CTLA-4 118:123 CTLA-4 118:123 CTLA-4 118:123 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 1 60 theme activation-induced 131:148 arg1 receptor 174:181 an activation-induced, homodimeric inhibitory receptor 128:181 an activation-induced, homodimeric inhibitory receptor in T cells 128:192 CTLA-4 is an activation-induced, homodimeric inhibitory receptor in T cells. 16002699 0 61 theme biological 72:81 arg1 relevance 83:91 its biological relevance 68:91 its biological relevance for T cell inactivation 68:115 Hierarchical regulation of CTLA-4 dimer-based lattice formation and its biological relevance for T cell inactivation. 16002699 2 62 theme Recent 195:200 arg1 reports 219:225 Recent crystallographic reports 195:225 Recent crystallographic reports 195:225 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 2 63 theme lattice-like 259:270 arg1 arrays 272:277 lattice-like arrays 259:277 lattice-like arrays on the cell surface 259:297 Recent crystallographic reports have suggested that it may form lattice-like arrays on the cell surface upon binding B7.1/B7.2 (CD80, CD86) molecules. 16002699 4 64 theme CTLA-4 586:591 arg1 % 575:575 up to 35% 567:575 up to 35% of C122A CTLA-4 567:591 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 4 64 theme CTLA-4 586:591 arg1 CTLA-4 586:591 C122A CTLA-4 580:591 C122A CTLA-4 580:591 Surprisingly, we found that up to 35% of C122A CTLA-4 dimerized in human T lymphocytes. 16002699 3 65 theme point 430:434 arg1 mutation 436:443 a C122A point mutation 422:443 a C122A point mutation 422:443 To test the biological relevance of these CTLA-4-B7 lattices, we introduced a C122A point mutation in human CTLA-4, because this residue was shown to be essential for dimerization in solution. 16002699 8 66 theme mutant 1132:1137 arg1 molecules 1139:1147 these mutant molecules 1126:1147 these mutant molecules 1126:1147 Such functionality correlated with B7-induced dimerization of these mutant molecules. 16002699 6 67 theme glycosylation 881:893 arg1 sites 895:899 the N78 and N110 glycosylation sites 864:899 sites 895:899 C122-independent dimerization of CTLA-4 involved N-glycosylation, because further mutation of the N78 and N110 glycosylation sites abrogated dimerization. 15385547 8 0 theme embedded 1230:1237 arg1 domain 1223:1228 a catalytic domain 1211:1228 a catalytic domain embedded in the membrane in opposite orientation to that of presenilins 1211:1300 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 5 1 contain contain 807:813 arg1 SPP 793:795 SPP 793:795 SPP 793:795 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 5 1 contain contain 807:813 arg2 sequence 838:845 a type I signal anchor sequence 815:845 a type I signal anchor sequence for initiation of protein translocation and membrane insertion 815:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 5 1 contain contain 807:813 arg1 SPPL3 801:805 SPPL3 801:805 SPPL3 801:805 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 1 2 theme human 165:169 arg1 proteases 229:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 1 2 theme human 165:169 arg1 genome 171:176 The human genome 161:176 The human genome 161:176 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 0 3 theme proteases 77:85 arg1 analysis 10:17 Consensus analysis 0:17 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases 0:85 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 6 4 theme catalytic 986:994 arg1 residues 996:1003 the catalytic residues 982:1003 the catalytic residues 982:1003 The hydrophilic loops joining the transmembrane regions, which contain the catalytic residues, are facing the exoplasm. 15385547 5 5 theme type 817:820 arg1 sequence 838:845 a type I signal anchor sequence 815:845 a type I signal anchor sequence for initiation of protein translocation and membrane insertion 815:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 3 6 theme comparative 500:510 arg1 analysis 512:519 a comparative analysis 498:519 a comparative analysis of the topologies of SPP 498:544 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 4 7 from located 650:656 arg1 space 679:683 the extracellular space 661:683 the extracellular space 661:683 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 9 8 theme substrate 1403:1411 arg1 peptides 1413:1420 type II-oriented substrate peptides 1386:1420 type II-oriented substrate peptides 1386:1420 Other than presenilins, SPPL2a, -2b, -2c, and -3 are therefore predicted to cleave type II-oriented substrate peptides like the prototypic protease SPP. 15385547 7 9 theme C 1035:1035 arg1 termini 1037:1043 The C termini 1031:1043 The C termini of all these proteins 1031:1065 The C termini of all these proteins are exposed toward the cytosol. 15385547 9 10 theme protease 1442:1449 arg1 SPP 1451:1453 the prototypic protease SPP 1427:1453 the prototypic protease SPP 1427:1453 Other than presenilins, SPPL2a, -2b, -2c, and -3 are therefore predicted to cleave type II-oriented substrate peptides like the prototypic protease SPP. 15385547 5 11 theme translocation 873:885 arg1 initiation 851:860 initiation 851:860 initiation of protein translocation and membrane insertion 851:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 6 12 theme hydrophilic 915:925 arg1 loops 927:931 The hydrophilic loops 911:931 The hydrophilic loops joining the transmembrane regions, which contain the catalytic residues, 911:1004 The hydrophilic loops joining the transmembrane regions, which contain the catalytic residues, are facing the exoplasm. 15385547 0 13 theme opposite 95:102 arg1 topology 104:111 opposite topology 95:111 opposite topology of catalytic domains 95:132 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 5 14 theme membrane 891:898 arg1 insertion 900:908 membrane insertion 891:908 membrane insertion 891:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 3 15 theme SPP 542:544 arg1 topologies 528:537 the topologies 524:537 the topologies of SPP 524:544 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 9 16 theme type 1386:1389 arg1 peptides 1413:1420 type II-oriented substrate peptides 1386:1420 type II-oriented substrate peptides 1386:1420 Other than presenilins, SPPL2a, -2b, -2c, and -3 are therefore predicted to cleave type II-oriented substrate peptides like the prototypic protease SPP. 15385547 0 17 theme catalytic 116:124 arg1 domains 126:132 catalytic domains 116:132 catalytic domains 116:132 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 4 18 located located 650:656 arg2 extensions 635:644 their N-terminal extensions 618:644 their N-terminal extensions 618:644 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 4 18 located located 650:656 arg1 space 679:683 the extracellular space 661:683 the extracellular space 661:683 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 8 19 theme principal 1186:1194 arg1 structure 1196:1204 the same principal structure 1177:1204 the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins 1177:1300 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 4 20 dep located 650:656 arg1 modified 712:719 modified 712:719 are modified with N-glycans 708:734 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 1 21 theme intramembrane-cleaving 192:213 arg1 proteases 229:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 1 21 theme intramembrane-cleaving 192:213 arg1 genome 171:176 The human genome 161:176 The human genome 161:176 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 0 22 theme peptidase 37:45 arg1 analysis 10:17 Consensus analysis 0:17 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases 0:85 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 3 23 theme topologies 528:537 arg1 -3 594:595 -3 594:595 -3 594:595 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 23 theme topologies 528:537 arg1 -2b 580:582 -2b 580:582 -2b 580:582 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 23 theme topologies 528:537 arg1 analysis 512:519 a comparative analysis 498:519 a comparative analysis of the topologies of SPP 498:544 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 23 theme topologies 528:537 arg1 homologues 560:569 its human homologues 550:569 its human homologues 550:569 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 23 theme topologies 528:537 arg1 -2c 585:587 -2c 585:587 -2c 585:587 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 23 theme topologies 528:537 arg1 SPPL2a 572:577 SPPL2a 572:577 SPPL2a 572:577 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 8 24 from embedded 1230:1237 arg1 membrane 1246:1253 the membrane 1242:1253 the membrane in opposite orientation to that of presenilins 1242:1300 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 3 25 theme human 554:558 arg1 -2c 585:587 -2c 585:587 -2c 585:587 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 25 theme human 554:558 arg1 -3 594:595 -3 594:595 -3 594:595 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 25 theme human 554:558 arg1 -2b 580:582 -2b 580:582 -2b 580:582 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 25 theme human 554:558 arg1 SPPL2a 572:577 SPPL2a 572:577 SPPL2a 572:577 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 3 25 theme human 554:558 arg1 homologues 560:569 its human homologues 550:569 its human homologues 550:569 Here we describe a comparative analysis of the topologies of SPP and its human homologues, SPPL2a, -2b, -2c, and -3. 15385547 6 26 contain contain 974:980 arg1 regions 959:965 the transmembrane regions 941:965 the transmembrane regions 941:965 The hydrophilic loops joining the transmembrane regions, which contain the catalytic residues, are facing the exoplasm. 15385547 6 26 contain contain 974:980 arg2 residues 996:1003 the catalytic residues 982:1003 the catalytic residues 982:1003 The hydrophilic loops joining the transmembrane regions, which contain the catalytic residues, are facing the exoplasm. 15385547 2 27 theme immune 392:397 arg1 surveillance 399:410 immune surveillance 392:410 immune surveillance 392:410 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 0 28 theme Consensus 0:8 arg1 analysis 10:17 Consensus analysis 0:17 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases 0:85 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 8 29 theme opposite 1258:1265 arg1 orientation 1267:1277 opposite orientation 1258:1277 opposite orientation 1258:1277 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 1 30 theme GXGD 215:218 arg1 proteases 229:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 1 30 theme GXGD 215:218 arg1 genome 171:176 The human genome 161:176 The human genome 161:176 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 0 31 theme signal 22:27 arg1 peptidase 37:45 signal peptide peptidase 22:45 signal peptide peptidase 22:45 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 5 32 contain contain 766:772 arg1 SPPL2a 745:750 SPPL2a 745:750 SPPL2a 745:750 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 5 32 contain contain 766:772 arg2 sequence 783:790 a signal sequence 774:790 a signal sequence 774:790 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 5 32 contain contain 766:772 arg1 -2b 753:755 -2b 753:755 -2b 753:755 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 5 32 contain contain 766:772 arg1 -2c 762:764 -2c 762:764 -2c 762:764 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 5 33 theme protein 865:871 arg1 translocation 873:885 protein translocation 865:885 protein translocation 865:885 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 2 34 theme candidate 431:439 arg1 SPPLs 452:456 SPPLs 452:456 SPPLs 452:456 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 34 theme candidate 431:439 arg1 proteases 441:449 four SPP-like candidate proteases 417:449 four SPP-like candidate proteases (SPPLs) 417:457 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 0 35 theme domains 126:132 arg1 topology 104:111 opposite topology 95:111 opposite topology of catalytic domains 95:132 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 5 36 theme I 822:822 arg1 sequence 838:845 a type I signal anchor sequence 815:845 a type I signal anchor sequence for initiation of protein translocation and membrane insertion 815:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 2 37 theme SPP-like 422:429 arg1 SPPLs 452:456 SPPLs 452:456 SPPLs 452:456 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 37 theme SPP-like 422:429 arg1 proteases 441:449 four SPP-like candidate proteases 417:449 four SPP-like candidate proteases (SPPLs) 417:457 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 38 theme function 471:478 arg1 peptidase 362:370 signal peptide peptidase 347:370 signal peptide peptidase (SPP) 347:376 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 38 theme function 471:478 arg1 SPPLs 452:456 SPPLs 452:456 SPPLs 452:456 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 38 theme function 471:478 arg1 proteases 441:449 four SPP-like candidate proteases 417:449 four SPP-like candidate proteases (SPPLs) 417:457 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 38 theme function 471:478 arg1 disease 338:344 Alzheimer's disease 326:344 Alzheimer's disease 326:344 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 38 theme function 471:478 arg1 SPP 373:375 SPP 373:375 SPP 373:375 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 5 39 theme anchor 831:836 arg1 sequence 838:845 a type I signal anchor sequence 815:845 a type I signal anchor sequence for initiation of protein translocation and membrane insertion 815:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 8 40 theme same 1181:1184 arg1 structure 1196:1204 the same principal structure 1177:1204 the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins 1177:1300 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 4 41 theme N-terminal 624:633 arg1 extensions 635:644 their N-terminal extensions 618:644 their N-terminal extensions 618:644 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 5 42 theme signal 776:781 arg1 sequence 783:790 a signal sequence 774:790 a signal sequence 774:790 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 0 43 theme peptide 29:35 arg1 peptidase 37:45 signal peptide peptidase 22:45 signal peptide peptidase 22:45 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 2 44 theme unknown 463:469 arg1 function 471:478 unknown function 463:478 unknown function 463:478 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 8 45 with structure 1196:1204 arg1 domain 1223:1228 a catalytic domain 1211:1228 a catalytic domain embedded in the membrane in opposite orientation to that of presenilins 1211:1300 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 0 46 theme homologous 51:60 arg1 proteases 77:85 homologous human aspartic proteases 51:85 homologous human aspartic proteases 51:85 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 5 47 theme signal 824:829 arg1 sequence 838:845 a type I signal anchor sequence 815:845 a type I signal anchor sequence for initiation of protein translocation and membrane insertion 815:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 7 48 theme proteins 1058:1065 arg1 termini 1037:1043 The C termini 1031:1043 The C termini of all these proteins 1031:1065 The C termini of all these proteins are exposed toward the cytosol. 15385547 4 49 theme extracellular 665:677 arg1 space 679:683 the extracellular space 661:683 the extracellular space 661:683 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 9 50 theme prototypic 1431:1440 arg1 SPP 1451:1453 the prototypic protease SPP 1427:1453 the prototypic protease SPP 1427:1453 Other than presenilins, SPPL2a, -2b, -2c, and -3 are therefore predicted to cleave type II-oriented substrate peptides like the prototypic protease SPP. 15385547 2 51 theme peptide 354:360 arg1 peptidase 362:370 signal peptide peptidase 347:370 signal peptide peptidase (SPP) 347:376 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 51 theme peptide 354:360 arg1 SPP 373:375 SPP 373:375 SPP 373:375 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 52 theme signaling 284:292 arg1 molecules 294:302 signaling molecules 284:302 signaling molecules 284:302 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 53 theme signal 347:352 arg1 peptidase 362:370 signal peptide peptidase 347:370 signal peptide peptidase (SPP) 347:376 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 2 53 theme signal 347:352 arg1 SPP 373:375 SPP 373:375 SPP 373:375 These are the two presenilins that activate signaling molecules and are implicated in Alzheimer's disease, signal peptide peptidase (SPP), required for immune surveillance, and four SPP-like candidate proteases (SPPLs), of unknown function. 15385547 4 54 from space 679:683 arg1 located 650:656 located 650:656 located 650:656 We demonstrate that their N-terminal extensions are located in the extracellular space and, except for SPPL3, are modified with N-glycans. 15385547 0 55 theme aspartic 68:75 arg1 proteases 77:85 homologous human aspartic proteases 51:85 homologous human aspartic proteases 51:85 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 5 56 theme insertion 900:908 arg1 initiation 851:860 initiation 851:860 initiation of protein translocation and membrane insertion 851:908 Whereas SPPL2a, -2b, and -2c contain a signal sequence, SPP and SPPL3 contain a type I signal anchor sequence for initiation of protein translocation and membrane insertion. 15385547 1 57 theme aspartic 220:227 arg1 proteases 229:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 seven intramembrane-cleaving GXGD aspartic proteases 186:237 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 1 57 theme aspartic 220:227 arg1 genome 171:176 The human genome 161:176 The human genome 161:176 The human genome encodes seven intramembrane-cleaving GXGD aspartic proteases. 15385547 9 58 theme II-oriented 1391:1401 arg1 peptides 1413:1420 type II-oriented substrate peptides 1386:1420 type II-oriented substrate peptides 1386:1420 Other than presenilins, SPPL2a, -2b, -2c, and -3 are therefore predicted to cleave type II-oriented substrate peptides like the prototypic protease SPP. 15385547 6 59 theme transmembrane 945:957 arg1 regions 959:965 the transmembrane regions 941:965 the transmembrane regions 941:965 The hydrophilic loops joining the transmembrane regions, which contain the catalytic residues, are facing the exoplasm. 15385547 0 60 theme human 62:66 arg1 proteases 77:85 homologous human aspartic proteases 51:85 homologous human aspartic proteases 51:85 Consensus analysis of signal peptide peptidase and homologous human aspartic proteases reveals opposite topology of catalytic domains compared with presenilins. 15385547 8 61 from membrane 1246:1253 arg1 embedded 1230:1237 embedded 1230:1237 embedded 1230:1237 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 8 61 from membrane 1246:1253 arg1 orientation 1267:1277 opposite orientation 1258:1277 opposite orientation 1258:1277 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 15385547 8 62 theme catalytic 1213:1221 arg1 domain 1223:1228 a catalytic domain 1211:1228 a catalytic domain embedded in the membrane in opposite orientation to that of presenilins 1211:1300 Taken together, our study demonstrates that SPP and its homologues are all of the same principal structure with a catalytic domain embedded in the membrane in opposite orientation to that of presenilins. 18467335 9 0 theme PCI 1113:1115 arg1 fraction 1101:1108 a small fraction 1093:1108 a small fraction of PCI 1093:1115 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 2 1 theme anticoagulant 302:314 arg1 proteases 316:324 anticoagulant proteases 302:324 anticoagulant proteases 302:324 In blood, it is capable of inhibiting both procoagulant and anticoagulant proteases. 18467335 4 2 link blood-derived 503:515 arg1 PCI 517:519 blood-derived PCI 503:519 blood-derived PCI 503:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 11 3 theme thrombomodulin-enhanced 1384:1406 arg1 rates 1408:1412 the thrombomodulin-enhanced rates 1380:1412 the thrombomodulin-enhanced rates of thrombin inhibition 1380:1435 In contrast, the Delta6-N-region increased both the heparin- and the thrombomodulin-enhanced rates of thrombin inhibition at all conditions examined. 18467335 5 4 theme N-glycan 648:655 arg1 structures 657:666 N-glycan structures 648:666 N-glycan structures 648:666 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 6 5 theme complex 772:778 arg1 N-glycans 780:788 tetra-antennary complex N-glycans 756:788 tetra-antennary complex N-glycans 756:788 Bi-, tri-, and tetra-antennary complex N-glycans were identified. 18467335 1 6 theme serine 142:147 arg1 inhibitor 158:166 a serine protease inhibitor 140:166 a serine protease inhibitor 140:166 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 6 theme serine 142:147 arg1 inhibitor 121:129 Protein C inhibitor 111:129 Protein C inhibitor (PCI) 111:135 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 13 7 from association 1782:1792 arg1 states 1893:1898 reducing hypercoagulability states 1865:1898 reducing hypercoagulability states 1865:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 7 8 theme Fucose 807:812 arg1 parts 870:874 parts 870:874 parts of sialyl-Le(a/x) epitopes 870:901 Fucose residues were identified both on the core GlcNAc and as parts of sialyl-Le(a/x) epitopes. 18467335 7 8 theme Fucose 807:812 arg1 residues 814:821 Fucose residues 807:821 Fucose residues 807:821 Fucose residues were identified both on the core GlcNAc and as parts of sialyl-Le(a/x) epitopes. 18467335 13 9 with effect 1848:1853 arg1 plaques 1822:1828 atherosclerotic plaques 1806:1828 atherosclerotic plaques 1806:1828 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 1 10 theme protease 186:193 arg1 specificity 195:205 broad protease specificity 180:205 broad protease specificity 180:205 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 12 11 link N-linked 1505:1512 arg1 glycans 1514:1520 the N-linked glycans 1501:1520 the N-linked glycans 1501:1520 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 1 12 theme Protein 111:117 arg1 PCI 132:134 PCI 132:134 PCI 132:134 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 12 theme Protein 111:117 arg1 inhibitor 121:129 Protein C inhibitor 111:129 Protein C inhibitor (PCI) 111:135 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 12 theme Protein 111:117 arg1 inhibitor 158:166 a serine protease inhibitor 140:166 a serine protease inhibitor 140:166 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 10 13 theme overall 1153:1159 arg1 removal 1161:1167 The overall removal 1149:1167 The overall removal of N-glycans 1149:1180 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 0 14 theme inhibitor 42:50 arg1 N-glycans 0:8 N-glycans 0:8 N-glycans 0:8 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 0 14 theme inhibitor 42:50 arg1 terminus 20:27 the N terminus 14:27 the N terminus of protein C inhibitor 14:50 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 4 15 gly heterogeneity 486:498 arg1 PCI 517:519 blood-derived PCI 503:519 blood-derived PCI 503:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 13 16 theme documented 1771:1780 arg1 association 1782:1792 the documented association 1767:1792 the documented association of PCI with atherosclerotic plaques 1767:1828 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 4 17 theme full 449:452 arg1 explanation 454:464 a full explanation 447:464 a full explanation for the marked size heterogeneity of blood-derived PCI 447:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 5 18 gly occupancy 685:693 arg2 form 735:738 a Delta6-N-cleaved form 716:738 a Delta6-N-cleaved form 716:738 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 10 19 theme maximal 1195:1201 arg1 heparin- 1203:1210 maximal heparin- 1195:1210 maximal heparin- 1195:1210 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 13 20 from view 1759:1762 arg1 findings 1722:1729 The findings 1718:1729 The findings 1718:1729 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 13 20 from view 1759:1762 arg1 important 1745:1753 important 1745:1753 important 1745:1753 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 13 20 from view 1759:1762 arg1 states 1893:1898 reducing hypercoagulability states 1865:1898 reducing hypercoagulability states 1865:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 12 21 from region 1541:1546 arg1 ways 1582:1585 different ways 1572:1585 different ways 1572:1585 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 10 22 dep heparin- 1203:1210 arg1 the 1191:1193 the 1191:1193 the 1191:1193 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 13 23 theme promising 1838:1846 arg1 effect 1848:1853 the promising effect 1834:1853 the promising effect of PCI on reducing hypercoagulability states 1834:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 4 24 theme first 427:431 arg1 time 433:436 the first time 423:436 the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants 423:598 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 11 25 theme inhibition 1426:1435 arg1 rates 1408:1412 the thrombomodulin-enhanced rates 1380:1412 the thrombomodulin-enhanced rates of thrombin inhibition 1380:1435 In contrast, the Delta6-N-region increased both the heparin- and the thrombomodulin-enhanced rates of thrombin inhibition at all conditions examined. 18467335 11 25 theme inhibition 1426:1435 arg1 heparin- 1367:1374 the heparin- 1363:1374 the heparin- 1363:1374 In contrast, the Delta6-N-region increased both the heparin- and the thrombomodulin-enhanced rates of thrombin inhibition at all conditions examined. 18467335 5 26 theme form 735:738 arg1 structures 657:666 N-glycan structures 648:666 N-glycan structures 648:666 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 5 26 theme form 735:738 arg1 occupancy 685:693 N-glycosylation occupancy 669:693 N-glycosylation occupancy 669:693 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 5 26 theme form 735:738 arg1 presence 704:711 the presence 700:711 the presence of a Delta6-N-cleaved form 700:738 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 9 27 gly N-glycosylated 997:1010 arg1 PCI 989:991 PCI 989:991 PCI 989:991 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 27 gly N-glycosylated 997:1010 arg2 Asn-243 1067:1073 Asn-243 1067:1073 Asn-243 1067:1073 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 27 gly N-glycosylated 997:1010 arg2 Asn-230 1058:1064 Asn-230 1058:1064 Asn-230 1058:1064 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 27 gly N-glycosylated 997:1010 arg2 Asn-319 1080:1086 Asn-319 1080:1086 Asn-319 1080:1086 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 27 gly N-glycosylated 997:1010 arg2 sites 1051:1055 all three potential N-glycosylation sites 1015:1055 all three potential N-glycosylation sites 1015:1055 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 13 28 from states 1893:1898 arg1 view 1759:1762 view 1759:1762 view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states 1759:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 13 29 theme reducing 1865:1872 arg1 states 1893:1898 reducing hypercoagulability states 1865:1898 reducing hypercoagulability states 1865:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 12 30 from glycans 1514:1520 arg1 ways 1582:1585 different ways 1572:1585 different ways 1572:1585 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 12 31 theme inhibition 1634:1643 arg1 rates 1616:1620 the cofactor-enhanced rates 1594:1620 the cofactor-enhanced rates of thrombin inhibition 1594:1643 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 4 32 theme blood-derived 503:515 arg1 PCI 517:519 blood-derived PCI 503:519 blood-derived PCI 503:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 4 33 dep time 433:436 arg1 provided 438:445 provided 438:445 provided a full explanation for the marked size heterogeneity of blood-derived PCI 438:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 4 33 dep time 433:436 arg1 identified 525:534 identified 525:534 identified functional differences between naturally occurring PCI variants 525:598 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 10 34 theme thrombin 1249:1256 arg1 inhibition 1258:1267 thrombin inhibition 1249:1267 thrombin inhibition 1249:1267 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 13 35 with association 1782:1792 arg1 plaques 1822:1828 atherosclerotic plaques 1806:1828 atherosclerotic plaques 1806:1828 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 13 36 theme effect 1848:1853 arg1 view 1759:1762 view 1759:1762 view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states 1759:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 12 37 theme N-linked 1505:1512 arg1 glycans 1514:1520 the N-linked glycans 1501:1520 the N-linked glycans 1501:1520 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 13 38 from effect 1848:1853 arg1 states 1893:1898 reducing hypercoagulability states 1865:1898 reducing hypercoagulability states 1865:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 4 39 theme marked 474:479 arg1 heterogeneity 486:498 the marked size heterogeneity 470:498 the marked size heterogeneity of blood-derived PCI 470:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 0 40 theme protein 32:38 arg1 inhibitor 42:50 protein C inhibitor 32:50 protein C inhibitor 32:50 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 1 41 located found 208:212 arg2 inhibitor 121:129 Protein C inhibitor 111:129 Protein C inhibitor (PCI) 111:135 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 41 located found 208:212 arg2 inhibitor 158:166 a serine protease inhibitor 140:166 a serine protease inhibitor 140:166 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 41 located found 208:212 arg1 blood 217:221 blood 217:221 blood 217:221 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 41 located found 208:212 arg1 tissues 233:239 other tissues 227:239 other tissues 227:239 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 9 42 theme at 1137:1138 arg1 Asn-243 1140:1146 the N-glycan at Asn-243 1124:1146 the N-glycan at Asn-243 1124:1146 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 43 from sites 1051:1055 arg1 N-glycosylated 997:1010 N-glycosylated 997:1010 N-glycosylated 997:1010 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 10 44 theme different 1284:1292 arg1 conditions 1303:1312 different solution conditions 1284:1312 different solution conditions 1284:1312 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 4 45 theme occurring 577:585 arg1 variants 591:598 naturally occurring PCI variants 567:598 naturally occurring PCI variants 567:598 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 13 46 theme PCI 1797:1799 arg1 association 1782:1792 the documented association 1767:1792 the documented association of PCI with atherosclerotic plaques 1767:1828 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 13 46 theme PCI 1797:1799 arg1 effect 1848:1853 the promising effect 1834:1853 the promising effect of PCI on reducing hypercoagulability states 1834:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 9 47 theme small 1095:1099 arg1 fraction 1101:1108 a small fraction 1093:1108 a small fraction of PCI 1093:1115 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 7 48 theme sialyl-Le 879:887 arg1 epitopes 894:901 sialyl-Le(a/x) epitopes 879:901 sialyl-Le(a/x) epitopes 879:901 Fucose residues were identified both on the core GlcNAc and as parts of sialyl-Le(a/x) epitopes. 18467335 12 49 theme blood-derived 1551:1563 arg1 PCI 1565:1567 blood-derived PCI 1551:1567 blood-derived PCI 1551:1567 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 13 50 theme atherosclerotic 1806:1820 arg1 plaques 1822:1828 atherosclerotic plaques 1806:1828 atherosclerotic plaques 1806:1828 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 4 51 theme functional 536:545 arg1 differences 547:557 functional differences 536:557 functional differences between naturally occurring PCI variants 536:598 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 1 52 theme other 227:231 arg1 tissues 233:239 other tissues 227:239 other tissues 227:239 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 9 53 theme N-glycosylation 1035:1049 arg1 sites 1051:1055 all three potential N-glycosylation sites 1015:1055 all three potential N-glycosylation sites 1015:1055 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 53 theme N-glycosylation 1035:1049 arg1 Asn-243 1067:1073 Asn-243 1067:1073 Asn-243 1067:1073 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 53 theme N-glycosylation 1035:1049 arg1 Asn-319 1080:1086 Asn-319 1080:1086 Asn-319 1080:1086 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 53 theme N-glycosylation 1035:1049 arg1 Asn-230 1058:1064 Asn-230 1058:1064 Asn-230 1058:1064 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 13 54 theme association 1782:1792 arg1 view 1759:1762 view 1759:1762 view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states 1759:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 9 55 theme N-glycan 1128:1135 arg1 Asn-243 1140:1146 the N-glycan at Asn-243 1124:1146 the N-glycan at Asn-243 1124:1146 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 7 56 theme core 851:854 arg1 GlcNAc 856:861 the core GlcNAc 847:861 the core GlcNAc 847:861 Fucose residues were identified both on the core GlcNAc and as parts of sialyl-Le(a/x) epitopes. 18467335 8 57 theme di-sialyl 957:965 arg1 antenna 967:973 a di-sialyl antenna 955:973 a di-sialyl antenna 955:973 Moreover, a glycan with a composition that implied a di-sialyl antenna was observed. 18467335 12 58 from information 1657:1667 arg1 mechanisms 1676:1685 the mechanisms 1672:1685 the mechanisms by which this may be achieved 1672:1715 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 6 59 theme tetra-antennary 756:770 arg1 N-glycans 780:788 tetra-antennary complex N-glycans 756:788 tetra-antennary complex N-glycans 756:788 Bi-, tri-, and tetra-antennary complex N-glycans were identified. 18467335 1 60 theme protease 149:156 arg1 inhibitor 158:166 a serine protease inhibitor 140:166 a serine protease inhibitor 140:166 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 60 theme protease 149:156 arg1 inhibitor 121:129 Protein C inhibitor 111:129 Protein C inhibitor (PCI) 111:135 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 12 61 theme cofactor-enhanced 1598:1614 arg1 rates 1616:1620 the cofactor-enhanced rates 1594:1620 the cofactor-enhanced rates of thrombin inhibition 1594:1643 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 1 62 theme broad 180:184 arg1 specificity 195:205 broad protease specificity 180:205 broad protease specificity 180:205 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 12 63 theme different 1572:1580 arg1 ways 1582:1585 different ways 1572:1585 different ways 1572:1585 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 10 64 theme N-glycans 1172:1180 arg1 removal 1161:1167 The overall removal 1149:1167 The overall removal of N-glycans 1149:1180 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 5 65 theme N-glycosylation 669:683 arg1 occupancy 685:693 N-glycosylation occupancy 669:693 N-glycosylation occupancy 669:693 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 1 66 theme C 119:119 arg1 PCI 132:134 PCI 132:134 PCI 132:134 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 66 theme C 119:119 arg1 inhibitor 121:129 Protein C inhibitor 111:129 Protein C inhibitor (PCI) 111:135 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 1 66 theme C 119:119 arg1 inhibitor 158:166 a serine protease inhibitor 140:166 a serine protease inhibitor 140:166 Protein C inhibitor (PCI) is a serine protease inhibitor, displaying broad protease specificity, found in blood and other tissues. 18467335 9 67 from Asn-243 1140:1146 arg1 N-glycan 1128:1135 the N-glycan at Asn-243 1124:1146 the N-glycan at Asn-243 1124:1146 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 67 from Asn-243 1140:1146 arg1 at 1137:1138 the N-glycan at Asn-243 1124:1146 the N-glycan at Asn-243 1124:1146 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 2 68 from blood 245:249 arg1 capable 258:264 capable 258:264 capable 258:264 In blood, it is capable of inhibiting both procoagulant and anticoagulant proteases. 18467335 12 69 theme PCI 1565:1567 arg1 glycans 1514:1520 the N-linked glycans 1501:1520 the N-linked glycans 1501:1520 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 12 69 theme PCI 1565:1567 arg1 region 1541:1546 the N-terminal region 1526:1546 the N-terminal region 1526:1546 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 13 70 theme hypercoagulability 1874:1891 arg1 states 1893:1898 reducing hypercoagulability states 1865:1898 reducing hypercoagulability states 1865:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 0 71 theme cofactor-enhanced 63:79 arg1 rates 81:85 the cofactor-enhanced rates 59:85 the cofactor-enhanced rates of thrombin inhibition 59:108 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 13 72 from important 1745:1753 arg1 view 1759:1762 view 1759:1762 view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states 1759:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 2 73 from capable 258:264 arg1 blood 245:249 blood 245:249 blood 245:249 In blood, it is capable of inhibiting both procoagulant and anticoagulant proteases. 18467335 12 74 theme N-terminal 1530:1539 arg1 region 1541:1546 the N-terminal region 1526:1546 the N-terminal region 1526:1546 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 12 75 link blood-derived 1551:1563 arg1 PCI 1565:1567 blood-derived PCI 1551:1567 blood-derived PCI 1551:1567 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 11 76 theme thrombin 1417:1424 arg1 inhibition 1426:1435 thrombin inhibition 1417:1435 thrombin inhibition 1417:1435 In contrast, the Delta6-N-region increased both the heparin- and the thrombomodulin-enhanced rates of thrombin inhibition at all conditions examined. 18467335 0 77 theme inhibition 99:108 arg1 rates 81:85 the cofactor-enhanced rates 59:85 the cofactor-enhanced rates of thrombin inhibition 59:108 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 9 78 theme potential 1025:1033 arg1 sites 1051:1055 all three potential N-glycosylation sites 1015:1055 all three potential N-glycosylation sites 1015:1055 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 78 theme potential 1025:1033 arg1 Asn-243 1067:1073 Asn-243 1067:1073 Asn-243 1067:1073 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 78 theme potential 1025:1033 arg1 Asn-319 1080:1086 Asn-319 1080:1086 Asn-319 1080:1086 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 78 theme potential 1025:1033 arg1 Asn-230 1058:1064 Asn-230 1058:1064 Asn-230 1058:1064 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 10 79 theme thrombomodulin-enhanced 1216:1238 arg1 rates 1240:1244 thrombomodulin-enhanced rates 1216:1244 thrombomodulin-enhanced rates 1216:1244 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 5 80 theme Delta6-N-cleaved 718:733 arg1 form 735:738 a Delta6-N-cleaved form 716:738 a Delta6-N-cleaved form 716:738 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 4 81 theme PCI 517:519 arg1 heterogeneity 486:498 the marked size heterogeneity 470:498 the marked size heterogeneity of blood-derived PCI 470:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 0 82 theme N 18:18 arg1 terminus 20:27 the N terminus 14:27 the N terminus of protein C inhibitor 14:50 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 9 83 gly N-glycosylation 1035:1049 arg2 three 1019:1023 three 1019:1023 three 1019:1023 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 83 gly N-glycosylation 1035:1049 arg2 Asn-243 1067:1073 Asn-243 1067:1073 Asn-243 1067:1073 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 83 gly N-glycosylation 1035:1049 arg2 Asn-230 1058:1064 Asn-230 1058:1064 Asn-230 1058:1064 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 83 gly N-glycosylation 1035:1049 arg2 Asn-319 1080:1086 Asn-319 1080:1086 Asn-319 1080:1086 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 9 83 gly N-glycosylation 1035:1049 arg2 sites 1051:1055 all three potential N-glycosylation sites 1015:1055 all three potential N-glycosylation sites 1015:1055 PCI was N-glycosylated at all three potential N-glycosylation sites, Asn-230, Asn-243, and Asn-319, but a small fraction of PCI lacked the N-glycan at Asn-243. 18467335 7 84 theme epitopes 894:901 arg1 parts 870:874 parts 870:874 parts of sialyl-Le(a/x) epitopes 870:901 Fucose residues were identified both on the core GlcNAc and as parts of sialyl-Le(a/x) epitopes. 18467335 7 84 theme epitopes 894:901 arg1 residues 814:821 Fucose residues 807:821 Fucose residues 807:821 Fucose residues were identified both on the core GlcNAc and as parts of sialyl-Le(a/x) epitopes. 18467335 12 85 theme thrombin 1625:1632 arg1 inhibition 1634:1643 thrombin inhibition 1625:1643 thrombin inhibition 1625:1643 These results thus demonstrate that the N-linked glycans and the N-terminal region of blood-derived PCI in different ways affect the cofactor-enhanced rates of thrombin inhibition and provide information on the mechanisms by which this may be achieved. 18467335 10 86 theme inhibition 1258:1267 arg1 rates 1240:1244 thrombomodulin-enhanced rates 1216:1244 thrombomodulin-enhanced rates 1216:1244 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 10 86 theme inhibition 1258:1267 arg1 heparin- 1203:1210 maximal heparin- 1195:1210 maximal heparin- 1195:1210 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 13 87 theme PCI 1858:1860 arg1 association 1782:1792 the documented association 1767:1792 the documented association of PCI with atherosclerotic plaques 1767:1828 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 13 87 theme PCI 1858:1860 arg1 effect 1848:1853 the promising effect 1834:1853 the promising effect of PCI on reducing hypercoagulability states 1834:1898 The findings are medically important, in view of the documented association of PCI with atherosclerotic plaques and the promising effect of PCI on reducing hypercoagulability states. 18467335 8 88 with glycan 916:921 arg1 composition 930:940 a composition 928:940 a composition that implied a di-sialyl antenna 928:973 Moreover, a glycan with a composition that implied a di-sialyl antenna was observed. 18467335 4 89 theme size 481:484 arg1 heterogeneity 486:498 the marked size heterogeneity 470:498 the marked size heterogeneity of blood-derived PCI 470:519 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 18467335 0 90 theme thrombin 90:97 arg1 inhibition 99:108 thrombin inhibition 90:108 thrombin inhibition 90:108 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 0 91 theme C 40:40 arg1 inhibitor 42:50 protein C inhibitor 32:50 protein C inhibitor 32:50 N-glycans and the N terminus of protein C inhibitor affect the cofactor-enhanced rates of thrombin inhibition. 18467335 10 92 theme solution 1294:1301 arg1 conditions 1303:1312 different solution conditions 1284:1312 different solution conditions 1284:1312 The overall removal of N-glycans affected the maximal heparin- and thrombomodulin-enhanced rates of thrombin inhibition differently in different solution conditions. 18467335 5 93 from differences 633:643 arg1 structures 657:666 N-glycan structures 648:666 N-glycan structures 648:666 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 5 93 from differences 633:643 arg1 occupancy 685:693 N-glycosylation occupancy 669:693 N-glycosylation occupancy 669:693 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 5 93 from differences 633:643 arg1 presence 704:711 the presence 700:711 the presence of a Delta6-N-cleaved form 700:738 The heterogeneity was caused by differences in N-glycan structures, N-glycosylation occupancy, and the presence of a Delta6-N-cleaved form. 18467335 4 94 theme PCI 587:589 arg1 variants 591:598 naturally occurring PCI variants 567:598 naturally occurring PCI variants 567:598 In this study we have for the first time provided a full explanation for the marked size heterogeneity of blood-derived PCI and identified functional differences between naturally occurring PCI variants. 15350125 2 0 theme adhering 496:503 arg1 configuration 482:494 a seven transmembrane (TM) domain configuration 448:494 a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule 448:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 5 1 theme crystal 1157:1163 arg1 structures 1165:1174 the crystal structures 1153:1174 the crystal structures of similar proteins 1153:1194 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 4 2 theme extramembranous 875:889 arg1 domains 891:897 the extramembranous domains 871:897 the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations 871:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 8 3 theme pathological 1479:1490 arg1 P290S 1502:1506 P290S 1502:1506 P290S 1502:1506 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 8 3 theme pathological 1479:1490 arg1 mutation 1492:1499 a pathological mutation 1477:1499 a pathological mutation (P290S) 1477:1507 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 2 4 theme ASBT 355:358 arg1 topology 343:350 The membrane topology 330:350 The membrane topology of ASBT 330:358 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 6 5 theme Simulated 1197:1205 arg1 annealing 1207:1215 Simulated annealing 1197:1215 Simulated annealing 1197:1215 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 0 6 theme domains 88:94 arg1 structure 49:57 putative three-dimensional structure 22:57 putative three-dimensional structure 22:57 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 6 theme domains 88:94 arg1 scanning 9:16 Topology scanning 0:16 Topology scanning 0:16 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 6 7 theme binding 1314:1320 arg1 sites 1322:1326 four distinct binding sites 1300:1326 four distinct binding sites 1300:1326 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 1 8 theme apical 164:169 arg1 transporter 198:208 The apical sodium-dependent bile acid transporter 160:208 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) 160:224 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 3 9 dep loops 688:692 arg1 3 700:700 3 700:700 3 700:700 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 3 9 dep loops 688:692 arg1 1 694:694 1 694:694 1 694:694 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 0 10 theme apical 103:108 arg1 SLC10A2 150:156 SLC10A2 150:156 SLC10A2 150:156 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 10 theme apical 103:108 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 7 11 theme predicted 1373:1381 arg1 domain 1391:1396 the predicted binding domain 1369:1396 the predicted binding domain 1369:1396 Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. 15350125 6 12 theme distinct 1305:1312 arg1 sites 1322:1326 four distinct binding sites 1300:1326 four distinct binding sites 1300:1326 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 6 13 theme protein 1275:1281 arg1 surface 1283:1289 the protein surface 1271:1289 the protein surface 1271:1289 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 3 14 theme Membrane 536:543 arg1 topology 545:552 Membrane topology 536:552 Membrane topology 536:552 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 1 15 theme bile 271:274 arg1 salts 276:280 bile salts 271:280 bile salts 271:280 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 2 16 theme inside 522:527 arg1 " 528:528 the "positive inside" 508:528 the "positive inside" rule 508:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 8 17 dep in 1540:1541 arg1 silico 1543:1548 silico 1543:1548 silico 1543:1548 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 1 18 theme salts 276:280 arg1 circulation 256:266 the enterohepatic circulation 238:266 the enterohepatic circulation of bile salts 238:280 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 0 19 theme bile 127:130 arg1 SLC10A2 150:156 SLC10A2 150:156 SLC10A2 150:156 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 19 theme bile 127:130 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 4 20 theme remote-threading 839:854 arg1 techniques 856:865 homology-modeling and remote-threading techniques 817:865 homology-modeling and remote-threading techniques 817:865 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 4 21 theme three-dimensional 764:780 arg1 model 782:786 a three-dimensional model 762:786 a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations 762:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 1 22 theme sodium-dependent 171:186 arg1 transporter 198:208 The apical sodium-dependent bile acid transporter 160:208 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) 160:224 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 0 23 theme sodium-dependent 110:125 arg1 SLC10A2 150:156 SLC10A2 150:156 SLC10A2 150:156 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 23 theme sodium-dependent 110:125 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 4 24 theme molecular 1022:1030 arg1 simulations 1041:1051 molecular dynamics simulations 1022:1051 molecular dynamics simulations 1022:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 8 25 theme available 1458:1466 arg1 data 1468:1471 available data 1458:1471 available data for a pathological mutation (P290S) 1458:1507 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 2 26 theme seven 450:454 arg1 transmembrane 456:468 transmembrane 456:468 transmembrane 456:468 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 1 27 theme bile 188:191 arg1 transporter 198:208 The apical sodium-dependent bile acid transporter 160:208 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) 160:224 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 2 28 theme transmembrane 456:468 arg1 configuration 482:494 a seven transmembrane (TM) domain configuration 448:494 a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule 448:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 4 29 theme energy 997:1002 arg1 minimizations 1004:1016 energy minimizations 997:1016 energy minimizations 997:1016 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 2 30 dep transmembrane 456:468 arg1 TM 471:472 TM 471:472 TM 471:472 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 4 31 theme 7TM 739:741 arg1 topology 743:750 a 7TM topology 737:750 a 7TM topology 737:750 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 1 32 theme acid 193:196 arg1 transporter 198:208 The apical sodium-dependent bile acid transporter 160:208 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) 160:224 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 0 33 theme Topology 0:7 arg1 scanning 9:16 Topology scanning 0:16 Topology scanning 0:16 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 8 34 theme bile 1588:1591 arg1 acids 1593:1597 bile acids 1588:1597 bile acids 1588:1597 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 4 35 theme techniques 856:865 arg1 approach 805:812 an approach 802:812 an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations 802:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 2 36 theme consensus 390:398 arg1 analysis 411:418 a consensus topography analysis 388:418 a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule 388:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 0 37 theme putative 22:29 arg1 structure 49:57 putative three-dimensional structure 22:57 putative three-dimensional structure 22:57 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 7 38 theme domain 1391:1396 arg1 mutagenesis 1354:1364 Subsequent site-directed mutagenesis 1329:1364 Subsequent site-directed mutagenesis of the predicted binding domain 1329:1396 Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. 15350125 0 39 theme transporter 137:147 arg1 domains 88:94 the extracellular binding domains 62:94 the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2) 62:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 39 theme transporter 137:147 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 8 40 theme in 1540:1541 arg1 mutagenesis 1550:1560 in silico mutagenesis 1540:1560 in silico mutagenesis 1540:1560 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 7 41 theme site-directed 1340:1352 arg1 mutagenesis 1354:1364 Subsequent site-directed mutagenesis 1329:1364 Subsequent site-directed mutagenesis of the predicted binding domain 1329:1396 Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. 15350125 7 42 theme Subsequent 1329:1338 arg1 mutagenesis 1354:1364 Subsequent site-directed mutagenesis 1329:1364 Subsequent site-directed mutagenesis of the predicted binding domain 1329:1396 Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. 15350125 0 43 theme acid 132:135 arg1 SLC10A2 150:156 SLC10A2 150:156 SLC10A2 150:156 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 43 theme acid 132:135 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 3 44 gly glycosylated 707:718 arg1 sites 643:647 reporter sites 634:647 reporter sites inserted in the putative extracellular loops 1 and 3 634:700 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 4 45 dep topology 743:750 arg1 the 724:726 the 724:726 the 724:726 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 4 45 dep topology 743:750 arg1 basis 728:732 basis 728:732 basis 728:732 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 2 46 theme domain 475:480 arg1 configuration 482:494 a seven transmembrane (TM) domain configuration 448:494 a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule 448:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 1 47 theme key 294:296 arg1 role 298:301 a key role 292:301 a key role 292:301 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 3 48 theme putative 665:672 arg1 loops 688:692 the putative extracellular loops 1 and 3 661:700 the putative extracellular loops 1 and 3 661:700 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 3 49 theme N-glycosylation-scanning 593:616 arg1 mutagenesis 618:628 N-glycosylation-scanning mutagenesis 593:628 N-glycosylation-scanning mutagenesis 593:628 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 0 50 theme three-dimensional 31:47 arg1 structure 49:57 putative three-dimensional structure 22:57 putative three-dimensional structure 22:57 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 5 51 theme similar 1179:1185 arg1 proteins 1187:1194 similar proteins 1179:1194 similar proteins 1179:1194 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 4 52 theme attachment 950:959 arg1 points 961:966 membrane attachment points 941:966 membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations 941:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 1 53 dep transporter 198:208 arg1 ASBT 211:214 ASBT 211:214 ASBT 211:214 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 1 53 dep transporter 198:208 arg1 SLC10A2 217:223 SLC10A2 217:223 SLC10A2 217:223 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 2 54 theme topography 400:409 arg1 analysis 411:418 a consensus topography analysis 388:418 a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule 388:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 2 55 theme positive 513:520 arg1 " 528:528 the "positive inside" 508:528 the "positive inside" rule 508:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 2 56 theme " 528:528 arg1 rule 530:533 the "positive inside" rule 508:533 the "positive inside" rule 508:533 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 4 57 theme homology-modeling 817:833 arg1 techniques 856:865 homology-modeling and remote-threading techniques 817:865 homology-modeling and remote-threading techniques 817:865 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 4 58 theme dynamics 1032:1039 arg1 simulations 1041:1051 molecular dynamics simulations 1022:1051 molecular dynamics simulations 1022:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 1 59 theme cholesterol 306:316 arg1 metabolism 318:327 cholesterol metabolism 306:327 cholesterol metabolism 306:327 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 15350125 4 60 theme membrane 941:948 arg1 points 961:966 membrane attachment points 941:966 membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations 941:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 5 61 theme other 1078:1082 arg1 indicators 1094:1103 other geometric indicators 1078:1103 other geometric indicators 1078:1103 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 6 62 theme natural 1247:1253 arg1 substrate 1255:1263 a natural substrate 1245:1263 a natural substrate 1245:1263 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 6 62 theme natural 1247:1253 arg1 acid 1239:1242 cholic acid 1232:1242 cholic acid 1232:1242 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 2 63 theme membrane 334:341 arg1 topology 343:350 The membrane topology 330:350 The membrane topology of ASBT 330:358 The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. 15350125 6 64 theme cholic 1232:1237 arg1 substrate 1255:1263 a natural substrate 1245:1263 a natural substrate 1245:1263 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 6 64 theme cholic 1232:1237 arg1 acid 1239:1242 cholic acid 1232:1242 cholic acid 1232:1242 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 5 65 theme geometric 1084:1092 arg1 indicators 1094:1103 other geometric indicators 1078:1103 other geometric indicators 1078:1103 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 3 66 theme reporter 634:641 arg1 sites 643:647 reporter sites 634:647 reporter sites inserted in the putative extracellular loops 1 and 3 634:700 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 5 67 theme Ramachandran 1054:1065 arg1 scores 1067:1072 Ramachandran scores 1054:1072 Ramachandran scores 1054:1072 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 3 68 theme extracellular 674:686 arg1 loops 688:692 the putative extracellular loops 1 and 3 661:700 the putative extracellular loops 1 and 3 661:700 Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. 15350125 8 69 theme mutant 1521:1526 arg1 model 1528:1532 the mutant model 1517:1532 the mutant model after in silico mutagenesis 1517:1560 This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. 15350125 0 70 theme binding 80:86 arg1 domains 88:94 the extracellular binding domains 62:94 the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2) 62:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 70 theme binding 80:86 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 5 71 from comparable 1128:1137 arg1 quality 1142:1148 quality 1142:1148 quality 1142:1148 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 6 72 theme acid 1239:1242 arg1 docking 1221:1227 docking 1221:1227 docking 1221:1227 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 6 72 theme acid 1239:1242 arg1 annealing 1207:1215 Simulated annealing 1197:1215 Simulated annealing 1197:1215 Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. 15350125 4 73 dep points 961:966 arg1 refined 983:989 refined 983:989 was refined using energy minimizations and molecular dynamics simulations 979:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 7 74 theme binding 1383:1389 arg1 domain 1391:1396 the predicted binding domain 1369:1396 the predicted binding domain 1369:1396 Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. 15350125 0 75 theme extracellular 66:78 arg1 domains 88:94 the extracellular binding domains 62:94 the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2) 62:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 0 75 theme extracellular 66:78 arg1 transporter 137:147 the apical sodium-dependent bile acid transporter 99:147 the apical sodium-dependent bile acid transporter (SLC10A2) 99:157 Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). 15350125 5 76 theme proteins 1187:1194 arg1 structures 1165:1174 the crystal structures 1153:1174 the crystal structures of similar proteins 1153:1194 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 4 77 theme ASBT 791:794 arg1 model 782:786 a three-dimensional model 762:786 a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations 762:1051 On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. 15350125 5 78 from quality 1142:1148 arg1 comparable 1128:1137 comparable 1128:1137 comparable 1128:1137 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 5 78 from quality 1142:1148 arg1 model 1119:1123 the model 1115:1123 the model 1115:1123 Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. 15350125 1 79 theme enterohepatic 242:254 arg1 circulation 256:266 the enterohepatic circulation 238:266 the enterohepatic circulation of bile salts 238:280 The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. 2668275 3 0 theme amino 312:316 arg1 acids 318:322 583 amino acids 308:322 583 amino acids 308:322 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 1 gly N-glycosylation 398:412 arg2 sites 414:418 four potential N-glycosylation sites 383:418 four potential N-glycosylation sites 383:418 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 1 gly N-glycosylation 398:412 arg2 four 383:386 four 383:386 four 383:386 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 1 gly N-glycosylation 398:412 arg1 residues 370:377 21 or 23 residues 361:377 21 or 23 residues 361:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 1 gly N-glycosylation 398:412 arg2 residues 370:377 21 or 23 residues 361:377 21 or 23 residues 361:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 9 2 theme oriented 986:993 arg1 residues 1018:1025 161 and 346 residues 1006:1025 161 and 346 residues 1006:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 9 2 theme oriented 986:993 arg1 domains 995:1001 Two glycosylated luminally oriented domains 959:1001 Two glycosylated luminally oriented domains of 161 and 346 residues 959:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 6 3 theme STS 610:612 arg1 resistance 596:605 The resistance 592:605 The resistance of STS toward proteinase K after translocation into microsomes 592:668 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 10 4 theme endoplasmic 1179:1189 arg1 reticulum 1191:1199 the endoplasmic reticulum 1175:1199 the endoplasmic reticulum 1175:1199 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 1 5 gly glycosylation 70:82 arg1 cells 124:128 BHK-21 cells 117:128 BHK-21 cells 117:128 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 5 6 theme membrane 574:581 arg1 protein 583:589 an integral membrane protein 562:589 an integral membrane protein 562:589 STS has the solubility properties of an integral membrane protein. 2668275 4 7 theme sites 448:452 arg1 Two 421:423 Two 421:423 Two 421:423 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 4 7 theme sites 448:452 arg1 sites 448:452 the N-glycosylation sites 428:452 the N-glycosylation sites 428:452 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 1 8 from topology 60:67 arg1 cells 124:128 BHK-21 cells 117:128 BHK-21 cells 117:128 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 11 9 with homology 1377:1384 arg1 A 1465:1465 arylsulfatase A 1451:1465 arylsulfatase A 1451:1465 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 9 with homology 1377:1384 arg1 B 1486:1486 arylsulfatase B 1472:1486 arylsulfatase B 1472:1486 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 9 with homology 1377:1384 arg1 sulfatases 1439:1448 the lysosomal sulfatases 1425:1448 the lysosomal sulfatases 1425:1448 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 12 10 theme receptor-independent 1660:1679 arg1 mechanism 1681:1689 a mannose 6-phosphate receptor-independent mechanism 1638:1689 a mannose 6-phosphate receptor-independent mechanism 1638:1689 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 4 11 theme asparagine 493:502 arg1 residues 504:511 the asparagine residues 47 and 259 489:522 the asparagine residues 47 and 259 489:522 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 12 12 with similarity 1505:1514 arg1 sulfatases 1541:1550 these two lysosomal sulfatases 1521:1550 these two lysosomal sulfatases 1521:1550 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 5 13 theme protein 583:589 arg1 properties 548:557 the solubility properties 533:557 the solubility properties of an integral membrane protein 533:589 STS has the solubility properties of an integral membrane protein. 2668275 12 14 contain contain 1566:1572 arg1 STS 1553:1555 STS 1553:1555 STS 1553:1555 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 12 14 contain contain 1566:1572 arg2 residues 1594:1601 mannose 6-phosphate residues 1574:1601 mannose 6-phosphate residues 1574:1601 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 12 15 theme mannose 1640:1646 arg1 mechanism 1681:1689 a mannose 6-phosphate receptor-independent mechanism 1638:1689 a mannose 6-phosphate receptor-independent mechanism 1638:1689 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 10 16 from reticulum 1191:1199 arg1 located 1150:1156 located 1150:1156 located 1150:1156 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 7 17 theme membrane-spanning 811:827 arg1 domains 829:835 two membrane-spanning domains 807:835 two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue 807:915 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 7 17 theme membrane-spanning 811:827 arg1 acids 844:848 amino acids 185-211 and 213-237 838:868 acids 844:848 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 7 17 theme membrane-spanning 811:827 arg1 213-237 862:868 213-237 862:868 213-237 862:868 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 3 18 theme N-terminal 332:341 arg1 peptide 350:356 an N-terminal signal peptide 329:356 an N-terminal signal peptide of 21 or 23 residues 329:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 6 19 theme microsomes 754:763 arg1 side 746:749 the luminal side 734:749 the luminal side of microsomes 734:763 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 9 20 gly glycosylated 963:974 arg1 residues 1018:1025 161 and 346 residues 1006:1025 161 and 346 residues 1006:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 9 20 gly glycosylated 963:974 arg1 domains 995:1001 Two glycosylated luminally oriented domains 959:1001 Two glycosylated luminally oriented domains of 161 and 346 residues 959:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 12 21 theme 6-phosphate 1648:1658 arg1 mechanism 1681:1689 a mannose 6-phosphate receptor-independent mechanism 1638:1689 a mannose 6-phosphate receptor-independent mechanism 1638:1689 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 3 22 theme signal 343:348 arg1 peptide 350:356 an N-terminal signal peptide 329:356 an N-terminal signal peptide of 21 or 23 residues 329:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 2 23 theme cDNA 146:149 arg1 clone 151:155 A 2.4-kilobase cDNA clone 131:155 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) 131:189 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) was isolated and sequenced, which encoded an enzymatically active protein. 2668275 3 24 theme potential 388:396 arg1 sites 414:418 four potential N-glycosylation sites 383:418 four potential N-glycosylation sites 383:418 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 24 theme potential 388:396 arg1 residues 370:377 21 or 23 residues 361:377 21 or 23 residues 361:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 7 25 theme proline 901:907 arg1 residue 909:915 a helix-breaking proline residue 884:915 a helix-breaking proline residue 884:915 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 12 26 theme mannose 1574:1580 arg1 residues 1594:1601 mannose 6-phosphate residues 1574:1601 mannose 6-phosphate residues 1574:1601 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 2 27 theme 2.4-kilobase 133:144 arg1 clone 151:155 A 2.4-kilobase cDNA clone 131:155 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) 131:189 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) was isolated and sequenced, which encoded an enzymatically active protein. 2668275 6 28 dep most 684:687 arg1 all 697:699 all 697:699 all 697:699 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 6 28 dep most 684:687 arg1 if 690:691 if 690:691 if 690:691 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 6 29 theme most 684:687 arg1 sequences 702:710 most, if not all, sequences 684:710 most, if not all, sequences of STS 684:717 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 11 30 theme domains 1405:1411 arg1 homology 1377:1384 the high homology 1368:1384 the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B 1368:1486 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 3 31 theme residues 370:377 arg1 sites 414:418 four potential N-glycosylation sites 383:418 four potential N-glycosylation sites 383:418 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 31 theme residues 370:377 arg1 residues 370:377 21 or 23 residues 361:377 21 or 23 residues 361:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 31 theme residues 370:377 arg1 peptide 350:356 an N-terminal signal peptide 329:356 an N-terminal signal peptide of 21 or 23 residues 329:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 11 32 theme STS 1333:1335 arg1 related 1357:1363 related 1357:1363 related 1357:1363 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 32 theme STS 1333:1335 arg1 stability 1320:1328 The stability 1316:1328 The stability of STS in lysosomes 1316:1348 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 6 33 theme luminal 738:744 arg1 side 746:749 the luminal side 734:749 the luminal side of microsomes 734:763 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 1 34 theme subcellular 89:99 arg1 distribution 101:112 subcellular distribution 89:112 subcellular distribution 89:112 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 10 35 theme multivesicular 1265:1278 arg1 endosomes 1280:1288 multivesicular endosomes 1265:1288 multivesicular endosomes 1265:1288 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 10 36 theme smaller 1202:1208 arg1 fractions 1210:1218 smaller fractions 1202:1218 smaller fractions 1202:1218 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 7 37 theme deduced 770:776 arg1 sequence 789:796 The deduced amino acid sequence 766:796 The deduced amino acid sequence 766:796 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 3 38 theme deduced 270:276 arg1 sequence 289:296 The deduced amino acid sequence 266:296 The deduced amino acid sequence 266:296 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 5 39 theme integral 565:572 arg1 protein 583:589 an integral membrane protein 562:589 an integral membrane protein 562:589 STS has the solubility properties of an integral membrane protein. 2668275 12 40 theme lysosomal 1531:1539 arg1 sulfatases 1541:1550 these two lysosomal sulfatases 1521:1550 these two lysosomal sulfatases 1521:1550 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 9 41 theme hydrophobic 1046:1056 arg1 domain 1058:1063 a hydrophobic domain 1044:1063 a hydrophobic domain spanning the membrane twice in opposite directions 1044:1114 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 3 42 theme amino 278:282 arg1 sequence 289:296 The deduced amino acid sequence 266:296 The deduced amino acid sequence 266:296 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 4 43 theme N-glycosylation 432:446 arg1 sites 448:452 the N-glycosylation sites 428:452 the N-glycosylation sites 428:452 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 10 44 theme cell 1251:1254 arg1 surface 1256:1262 the cell surface 1247:1262 the cell surface 1247:1262 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 7 45 theme amino 778:782 arg1 sequence 789:796 The deduced amino acid sequence 766:796 The deduced amino acid sequence 766:796 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 11 46 theme arylsulfatase 1472:1484 arg1 B 1486:1486 arylsulfatase B 1472:1486 arylsulfatase B 1472:1486 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 46 theme arylsulfatase 1472:1484 arg1 sulfatases 1439:1448 the lysosomal sulfatases 1425:1448 the lysosomal sulfatases 1425:1448 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 0 47 theme steroid-sulfatase 32:48 arg1 expression 12:21 expression 12:21 expression 12:21 Cloning and expression of human steroid-sulfatase. 2668275 0 47 theme steroid-sulfatase 32:48 arg1 Cloning 0:6 Cloning 0:6 Cloning 0:6 Cloning and expression of human steroid-sulfatase. 2668275 11 48 theme high 1372:1375 arg1 homology 1377:1384 the high homology 1368:1384 the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B 1368:1486 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 10 49 from located 1150:1156 arg1 reticulum 1191:1199 the endoplasmic reticulum 1175:1199 the endoplasmic reticulum 1175:1199 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 7 50 theme amino 838:842 arg1 domains 829:835 two membrane-spanning domains 807:835 two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue 807:915 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 7 50 theme amino 838:842 arg1 acids 844:848 amino acids 185-211 and 213-237 838:868 acids 844:848 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 9 51 theme residues 1018:1025 arg1 residues 1018:1025 161 and 346 residues 1006:1025 161 and 346 residues 1006:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 9 51 theme residues 1018:1025 arg1 domains 995:1001 Two glycosylated luminally oriented domains 959:1001 Two glycosylated luminally oriented domains of 161 and 346 residues 959:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 7 52 theme acid 784:787 arg1 sequence 789:796 The deduced amino acid sequence 766:796 The deduced amino acid sequence 766:796 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 0 53 theme human 26:30 arg1 steroid-sulfatase 32:48 human steroid-sulfatase 26:48 human steroid-sulfatase 26:48 Cloning and expression of human steroid-sulfatase. 2668275 4 54 dep residues 504:511 arg1 259 520:522 259 520:522 259 520:522 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 4 54 dep residues 504:511 arg1 47 513:514 47 513:514 47 513:514 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 3 55 theme N-glycosylation 398:412 arg1 sites 414:418 four potential N-glycosylation sites 383:418 four potential N-glycosylation sites 383:418 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 3 55 theme N-glycosylation 398:412 arg1 residues 370:377 21 or 23 residues 361:377 21 or 23 residues 361:377 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 8 56 theme three-domain 939:950 arg1 model 952:956 a three-domain model 937:956 a three-domain model 937:956 We propose for STS a three-domain model. 2668275 4 57 used utilized 458:465 arg2 sites 448:452 the N-glycosylation sites 428:452 the N-glycosylation sites 428:452 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 4 57 used utilized 458:465 arg2 Two 421:423 Two 421:423 Two 421:423 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 2 58 theme human 161:165 arg1 STS 186:188 STS 186:188 STS 186:188 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) was isolated and sequenced, which encoded an enzymatically active protein. 2668275 2 58 theme human 161:165 arg1 steroid-sulfatase 167:183 human steroid-sulfatase 161:183 human steroid-sulfatase (STS) 161:189 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) was isolated and sequenced, which encoded an enzymatically active protein. 2668275 1 59 theme BHK-21 117:122 arg1 cells 124:128 BHK-21 cells 117:128 BHK-21 cells 117:128 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 3 60 theme 583 308:310 arg1 acids 318:322 583 amino acids 308:322 583 amino acids 308:322 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 12 61 theme 6-phosphate 1582:1592 arg1 residues 1594:1601 mannose 6-phosphate residues 1574:1601 mannose 6-phosphate residues 1574:1601 In spite of its similarity with these two lysosomal sulfatases, STS does not contain mannose 6-phosphate residues and is transported to lysosomes by a mannose 6-phosphate receptor-independent mechanism. 2668275 9 62 theme opposite 1096:1103 arg1 directions 1105:1114 opposite directions 1096:1114 opposite directions 1096:1114 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 1 63 from distribution 101:112 arg1 cells 124:128 BHK-21 cells 117:128 BHK-21 cells 117:128 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 7 64 theme helix-breaking 886:899 arg1 residue 909:915 a helix-breaking proline residue 884:915 a helix-breaking proline residue 884:915 The deduced amino acid sequence predicts two membrane-spanning domains (amino acids 185-211 and 213-237) separated by a helix-breaking proline residue. 2668275 3 65 theme acid 284:287 arg1 sequence 289:296 The deduced amino acid sequence 266:296 The deduced amino acid sequence 266:296 The deduced amino acid sequence comprises 583 amino acids with an N-terminal signal peptide of 21 or 23 residues and four potential N-glycosylation sites. 2668275 11 66 from stability 1320:1328 arg1 lysosomes 1340:1348 lysosomes 1340:1348 lysosomes 1340:1348 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 5 67 theme solubility 537:546 arg1 properties 548:557 the solubility properties 533:557 the solubility properties of an integral membrane protein 533:589 STS has the solubility properties of an integral membrane protein. 2668275 5 68 contain has 529:531 arg2 properties 548:557 the solubility properties 533:557 the solubility properties of an integral membrane protein 533:589 STS has the solubility properties of an integral membrane protein. 2668275 5 68 contain has 529:531 arg1 STS 525:527 STS 525:527 STS 525:527 STS has the solubility properties of an integral membrane protein. 2668275 4 69 gly N-glycosylation 432:446 arg2 sites 448:452 the N-glycosylation sites 428:452 the N-glycosylation sites 428:452 Two of the N-glycosylation sites are utilized and were localized to the asparagine residues 47 and 259. 2668275 6 70 theme STS 715:717 arg1 sequences 702:710 most, if not all, sequences 684:710 most, if not all, sequences of STS 684:717 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 2 71 theme active 250:255 arg1 which 219:223 which 219:223 which 219:223 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) was isolated and sequenced, which encoded an enzymatically active protein. 2668275 2 71 theme active 250:255 arg1 protein 257:263 an enzymatically active protein 233:263 an enzymatically active protein 233:263 A 2.4-kilobase cDNA clone for human steroid-sulfatase (STS) was isolated and sequenced, which encoded an enzymatically active protein. 2668275 11 72 theme arylsulfatase 1451:1463 arg1 A 1465:1465 arylsulfatase A 1451:1465 arylsulfatase A 1451:1465 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 72 theme arylsulfatase 1451:1463 arg1 sulfatases 1439:1448 the lysosomal sulfatases 1425:1448 the lysosomal sulfatases 1425:1448 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 73 theme lysosomal 1429:1437 arg1 A 1465:1465 arylsulfatase A 1451:1465 arylsulfatase A 1451:1465 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 73 theme lysosomal 1429:1437 arg1 B 1486:1486 arylsulfatase B 1472:1486 arylsulfatase B 1472:1486 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 73 theme lysosomal 1429:1437 arg1 sulfatases 1439:1448 the lysosomal sulfatases 1425:1448 the lysosomal sulfatases 1425:1448 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 74 theme luminal 1397:1403 arg1 STS 1416:1418 STS 1416:1418 STS 1416:1418 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 74 theme luminal 1397:1403 arg1 domains 1405:1411 the two luminal domains 1389:1411 the two luminal domains of STS 1389:1418 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 75 theme STS 1416:1418 arg1 STS 1416:1418 STS 1416:1418 STS 1416:1418 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 11 75 theme STS 1416:1418 arg1 domains 1405:1411 the two luminal domains 1389:1411 the two luminal domains of STS 1389:1418 The stability of STS in lysosomes may be related to the high homology of the two luminal domains of STS with the lysosomal sulfatases, arylsulfatase A, and arylsulfatase B. 2668275 6 76 theme proteinase 621:630 arg1 K 632:632 proteinase K 621:632 proteinase K 621:632 The resistance of STS toward proteinase K after translocation into microsomes suggests that most, if not all, sequences of STS are exposed at the luminal side of microsomes. 2668275 9 77 theme glycosylated 963:974 arg1 residues 1018:1025 161 and 346 residues 1006:1025 161 and 346 residues 1006:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 9 77 theme glycosylated 963:974 arg1 domains 995:1001 Two glycosylated luminally oriented domains 959:1001 Two glycosylated luminally oriented domains of 161 and 346 residues 959:1025 Two glycosylated luminally oriented domains of 161 and 346 residues are separated by a hydrophobic domain spanning the membrane twice in opposite directions. 2668275 10 78 located found 1224:1228 arg1 surface 1256:1262 the cell surface 1247:1262 the cell surface 1247:1262 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 10 78 located found 1224:1228 arg1 Golgi 1237:1241 the Golgi 1233:1241 the Golgi 1233:1241 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 10 78 located found 1224:1228 arg2 fractions 1210:1218 smaller fractions 1202:1218 smaller fractions 1202:1218 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 2668275 1 79 theme Membrane 51:58 arg1 topology 60:67 Membrane topology 51:67 Membrane topology 51:67 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 1 80 from glycosylation 70:82 arg1 cells 124:128 BHK-21 cells 117:128 BHK-21 cells 117:128 Membrane topology, glycosylation, and subcellular distribution in BHK-21 cells. 2668275 10 81 theme BHK-21 1134:1139 arg1 cells 1141:1145 BHK-21 cells 1134:1145 BHK-21 cells 1134:1145 STS expressed in BHK-21 cells is located predominantly in the endoplasmic reticulum; smaller fractions are found in the Golgi, at the cell surface, multivesicular endosomes, as well as in lysosomes. 12214108 0 0 theme Site 64:67 arg1 Mutation 14:21 The Thr183Ala Mutation 0:21 The Thr183Ala Mutation 0:21 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 0 0 theme Site 64:67 arg1 Loss 32:35 Not the Loss 24:35 Not the Loss of the First Glycosylation Site 24:67 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 5 1 from residue 712:718 arg1 responsible 731:741 responsible 731:741 responsible 731:741 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 1 from residue 712:718 arg1 lack 681:684 the lack 677:684 the lack of the N-linked glycan at residue 181 677:722 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 0 2 theme Glycosylation 50:62 arg1 Site 64:67 the First Glycosylation Site 40:67 the First Glycosylation Site 40:67 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 1 3 theme abnormal 127:134 arg1 form 136:139 The abnormal form 123:139 The abnormal form of the prion protein 123:160 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 2 4 theme normal 255:260 arg1 protein 268:274 The normal prion protein 251:274 The normal prion protein 251:274 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 5 5 theme Ala 837:839 arg1 itself 853:858 itself 853:858 itself 853:858 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 5 theme Ala 837:839 arg1 mutation 841:848 the Thr to Ala mutation 826:848 the Thr to Ala mutation 826:848 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 1 6 theme non-ionic 229:237 arg1 detergents 239:248 non-ionic detergents 229:248 non-ionic detergents 229:248 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 0 7 theme Physical 81:88 arg1 Properties 90:99 the Physical Properties 77:99 the Physical Properties of the Prion Protein 77:120 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 5 8 dep itself 853:858 arg1 cause 873:877 the probable cause 860:877 itself the probable cause of the pathogenic phenotype 853:905 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 3 9 theme protein 414:420 arg1 residue 383:389 residue 183 383:393 residue 183 of the human prion protein 383:420 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 3 9 theme protein 414:420 arg1 protein 414:420 the human prion protein 398:420 the human prion protein 398:420 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 5 10 theme pathogen-like 792:804 arg1 properties 806:815 pathogen-like properties 792:815 pathogen-like properties 792:815 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 3 11 theme Asp 466:468 arg1 residue 470:476 the Asp residue 181 462:480 the Asp residue 181 462:480 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 0 12 theme Thr183Ala 4:12 arg1 Mutation 14:21 The Thr183Ala Mutation 0:21 The Thr183Ala Mutation 0:21 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 0 12 theme Thr183Ala 4:12 arg1 Loss 32:35 Not the Loss 24:35 Not the Loss of the First Glycosylation Site 24:67 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 2 13 link N-linked 326:333 arg1 glycans 335:341 two N-linked glycans 322:341 two N-linked glycans 322:341 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 4 14 theme glycan 654:659 arg1 lack 642:645 the lack 638:645 the lack of the glycan 638:659 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 4 15 theme pathogenic 597:606 arg1 properties 608:617 pathogenic properties 597:617 pathogenic properties 597:617 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 5 16 theme Thr 830:832 arg1 itself 853:858 itself 853:858 itself 853:858 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 16 theme Thr 830:832 arg1 mutation 841:848 the Thr to Ala mutation 826:848 the Thr to Ala mutation 826:848 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 1 17 theme prion 148:152 arg1 protein 154:160 the prion protein 144:160 the prion protein 144:160 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 5 18 dep mutation 841:848 arg1 to 834:835 to 834:835 to 834:835 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 2 19 mod modified 279:286 arg1 protein 268:274 The normal prion protein 251:274 The normal prion protein 251:274 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 2 19 mod modified 279:286 arg3 addition 310:317 the non-obligatory addition 291:317 the non-obligatory addition of two N-linked glycans 291:341 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 5 20 theme pathogenic 886:895 arg1 phenotype 897:905 the pathogenic phenotype 882:905 the pathogenic phenotype 882:905 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 1 21 theme protein 154:160 arg1 form 136:139 The abnormal form 123:139 The abnormal form of the prion protein 123:160 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 3 22 from residue 383:389 arg1 Thr 369:371 Thr 369:371 Thr to Ala at residue 183 of the human prion protein 369:420 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 3 22 from residue 383:389 arg1 mutation 359:366 One pathogenic mutation 344:366 One pathogenic mutation 344:366 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 2 23 theme prion 262:266 arg1 protein 268:274 The normal prion protein 251:274 The normal prion protein 251:274 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 3 24 theme glycan 452:457 arg1 addition 430:437 addition 430:437 addition of the first glycan to the Asp residue 181 430:480 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 0 25 theme Prion 108:112 arg1 Protein 114:120 the Prion Protein 104:120 the Prion Protein 104:120 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 5 26 link N-linked 693:700 arg1 glycan 702:707 the N-linked glycan 689:707 the N-linked glycan 689:707 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 1 27 theme increased 166:174 arg1 resistance 176:185 increased resistance 166:185 increased resistance to protease digestion 166:207 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 1 28 from insoluble 216:224 arg1 detergents 239:248 non-ionic detergents 229:248 non-ionic detergents 229:248 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 5 29 theme phenotype 897:905 arg1 cause 873:877 the probable cause 860:877 itself the probable cause of the pathogenic phenotype 853:905 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 3 30 theme human 402:406 arg1 protein 414:420 the human prion protein 398:420 the human prion protein 398:420 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 5 31 theme N-linked 693:700 arg1 glycan 702:707 the N-linked glycan 689:707 the N-linked glycan 689:707 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 32 theme properties 806:815 arg1 acquisition 777:787 the acquisition 773:787 the acquisition of pathogen-like properties 773:815 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 32 theme properties 806:815 arg1 block 751:755 the block 747:755 the block in transport 747:768 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 33 theme glycan 702:707 arg1 responsible 731:741 responsible 731:741 responsible 731:741 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 33 theme glycan 702:707 arg1 lack 681:684 the lack 677:684 the lack of the N-linked glycan at residue 181 677:722 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 0 34 theme Protein 114:120 arg1 Properties 90:99 the Physical Properties 77:99 the Physical Properties of the Prion Protein 77:120 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 3 35 theme first 446:450 arg1 glycan 452:457 the first glycan 442:457 the first glycan 442:457 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 2 36 theme non-obligatory 295:308 arg1 addition 310:317 the non-obligatory addition 291:317 the non-obligatory addition of two N-linked glycans 291:341 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 5 37 theme probable 864:871 arg1 cause 873:877 the probable cause 860:877 itself the probable cause of the pathogenic phenotype 853:905 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 5 38 from acquisition 777:787 arg1 transport 760:768 transport 760:768 transport 760:768 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 4 39 theme intracellular 528:540 arg1 retention 542:550 intracellular retention 528:550 intracellular retention of the mutant protein 528:572 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 1 40 contain has 162:164 arg2 resistance 176:185 increased resistance 166:185 increased resistance to protease digestion 166:207 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 1 40 contain has 162:164 arg1 form 136:139 The abnormal form 123:139 The abnormal form of the prion protein 123:160 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 4 41 theme properties 608:617 arg1 acquisition 582:592 its acquisition 578:592 its acquisition of pathogenic properties 578:617 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 4 41 theme properties 608:617 arg1 retention 542:550 intracellular retention 528:550 intracellular retention of the mutant protein 528:572 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 1 42 theme protease 190:197 arg1 digestion 199:207 protease digestion 190:207 protease digestion 190:207 The abnormal form of the prion protein has increased resistance to protease digestion and is insoluble in non-ionic detergents. 12214108 0 43 theme First 44:48 arg1 Site 64:67 the First Glycosylation Site 40:67 the First Glycosylation Site 40:67 The Thr183Ala Mutation, Not the Loss of the First Glycosylation Site, Alters the Physical Properties of the Prion Protein. 12214108 4 44 theme protein 566:572 arg1 acquisition 582:592 its acquisition 578:592 its acquisition of pathogenic properties 578:617 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 4 44 theme protein 566:572 arg1 retention 542:550 intracellular retention 528:550 intracellular retention of the mutant protein 528:572 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 3 45 theme prion 408:412 arg1 protein 414:420 the human prion protein 398:420 the human prion protein 398:420 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 2 46 theme glycans 335:341 arg1 addition 310:317 the non-obligatory addition 291:317 the non-obligatory addition of two N-linked glycans 291:341 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 12214108 3 47 theme pathogenic 348:357 arg1 Thr 369:371 Thr 369:371 Thr to Ala at residue 183 of the human prion protein 369:420 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 3 47 theme pathogenic 348:357 arg1 mutation 359:366 One pathogenic mutation 344:366 One pathogenic mutation 344:366 One pathogenic mutation, Thr to Ala at residue 183 of the human prion protein, blocks addition of the first glycan to the Asp residue 181. 12214108 4 48 theme mutant 559:564 arg1 protein 566:572 the mutant protein 555:572 the mutant protein 555:572 This mutation has been reported to result in intracellular retention of the mutant protein and its acquisition of pathogenic properties, presumably due to the lack of the glycan. 12214108 5 49 from block 751:755 arg1 transport 760:768 transport 760:768 transport 760:768 We report that the lack of the N-linked glycan at residue 181 is not responsible for the block in transport or the acquisition of pathogen-like properties, rather, the Thr to Ala mutation is itself the probable cause of the pathogenic phenotype. 12214108 2 50 theme N-linked 326:333 arg1 glycans 335:341 two N-linked glycans 322:341 two N-linked glycans 322:341 The normal prion protein is modified by the non-obligatory addition of two N-linked glycans. 22997138 3 0 theme form 489:492 arg1 structure 466:474 the crystal structure 454:474 the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity 454:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 0 1 theme ecto-5'-nucleotidase 55:74 arg1 activity 76:83 ecto-5'-nucleotidase activity 55:83 ecto-5'-nucleotidase activity 55:83 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 2 2 attach linked 350:355 arg1 cancer 404:409 cancer 404:409 cancer 404:409 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 2 attach linked 350:355 arg1 ischaemia 426:434 myocardial ischaemia 415:434 myocardial ischaemia 415:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 2 attach linked 350:355 arg2 CD73 188:191 CD73 188:191 CD73 188:191 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 2 attach linked 350:355 arg1 number 362:367 a number 360:367 a number of pathological conditions such as cancer and myocardial ischaemia 360:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 3 3 from form 489:492 arg1 resolution 533:542 2.2 Å resolution 527:542 2.2 Å resolution 527:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 1 4 theme exterior 150:157 arg1 side 159:162 the exterior side 146:162 the exterior side of the plasma membrane 146:185 CD73 is a dimeric ecto-5'-nucleotidase that is expressed on the exterior side of the plasma membrane. 22997138 2 5 from monophosphate 322:334 arg1 metabolism 249:258 the extracellular metabolism 231:258 the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, 231:335 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 7 6 theme sCD73 1148:1152 arg1 structure 1135:1143 The crystal structure 1123:1143 The crystal structure of sCD73 1123:1152 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 7 7 theme single 1302:1307 arg1 polymorphisms 1320:1332 single nucleotide polymorphisms 1302:1332 single nucleotide polymorphisms in physiology and disease 1302:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 7 8 theme nucleotide 1309:1318 arg1 polymorphisms 1320:1332 single nucleotide polymorphisms 1302:1332 single nucleotide polymorphisms in physiology and disease 1302:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 6 9 theme structures 1111:1120 arg1 variety 1084:1090 a variety 1082:1090 a variety of oligosaccharide structures 1082:1120 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 6 9 theme structures 1111:1120 arg1 structures 1111:1120 oligosaccharide structures 1095:1120 oligosaccharide structures 1095:1120 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 4 10 theme sense 717:721 arg1 dimerisation 663:674 the dimerisation 659:674 the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein 659:763 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 4 11 theme architecture 697:708 arg1 dimerisation 663:674 the dimerisation 659:674 the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein 659:763 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 2 12 theme monophosphates 282:295 arg1 metabolism 249:258 the extracellular metabolism 231:258 the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, 231:335 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 13 theme nucleoside 271:280 arg1 monophosphates 282:295 certain nucleoside monophosphates 263:295 certain nucleoside monophosphates 263:295 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 7 14 theme possible 1284:1291 arg1 roles 1293:1297 the possible roles 1280:1297 the possible roles of single nucleotide polymorphisms in physiology and disease 1280:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 6 15 theme enzyme 1032:1037 arg1 glycosylation 1011:1023 glycosylation 1011:1023 glycosylation of the enzyme 1011:1037 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 6 16 dep able 995:998 arg1 Using 950:954 Using 950:954 Using biophotonic microarray imaging 950:985 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 2 17 theme certain 263:269 arg1 monophosphates 282:295 certain nucleoside monophosphates 263:295 certain nucleoside monophosphates 263:295 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 18 theme conditions 385:394 arg1 cancer 404:409 cancer 404:409 cancer 404:409 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 18 theme conditions 385:394 arg1 ischaemia 426:434 myocardial ischaemia 415:434 myocardial ischaemia 415:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 18 theme conditions 385:394 arg1 number 362:367 a number 360:367 a number of pathological conditions such as cancer and myocardial ischaemia 360:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 6 19 theme oligosaccharide 1095:1109 arg1 structures 1111:1120 oligosaccharide structures 1095:1120 oligosaccharide structures 1095:1120 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 7 20 theme several 1232:1238 arg1 diseases 1240:1247 several diseases 1232:1247 several diseases 1232:1247 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 3 21 theme Å 531:531 arg1 resolution 533:542 2.2 Å resolution 527:542 2.2 Å resolution 527:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 2 22 theme pathological 372:383 arg1 conditions 385:394 pathological conditions 372:394 pathological conditions such as cancer and myocardial ischaemia 372:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 22 theme pathological 372:383 arg1 cancer 404:409 cancer 404:409 cancer 404:409 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 22 theme pathological 372:383 arg1 ischaemia 426:434 myocardial ischaemia 415:434 myocardial ischaemia 415:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 0 23 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 0:84 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 7 24 theme inhibitors 1177:1186 arg1 design 1167:1172 the design 1163:1172 the design of inhibitors or activator molecules 1163:1209 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 2 25 theme adenosine 312:320 arg1 monophosphate 322:334 particular adenosine monophosphate 301:334 particular adenosine monophosphate 301:334 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 1 26 theme plasma 171:176 arg1 membrane 178:185 the plasma membrane 167:185 the plasma membrane 167:185 CD73 is a dimeric ecto-5'-nucleotidase that is expressed on the exterior side of the plasma membrane. 22997138 5 27 theme crystal 770:776 arg1 structure 778:786 The crystal structure 766:786 The crystal structure 766:786 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 4 28 theme protein 757:763 arg1 modifications 736:748 secondary modifications 726:748 secondary modifications of the protein 726:763 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 5 29 theme dimer-dimer 846:856 arg1 interaction 858:868 the dimer-dimer interaction 842:868 the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges 842:947 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 2 30 theme particular 301:310 arg1 monophosphate 322:334 particular adenosine monophosphate 301:334 particular adenosine monophosphate 301:334 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 7 31 from polymorphisms 1320:1332 arg1 disease 1352:1358 disease 1352:1358 disease 1352:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 7 31 from polymorphisms 1320:1332 arg1 physiology 1337:1346 physiology 1337:1346 physiology 1337:1346 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 2 32 theme myocardial 415:424 arg1 ischaemia 426:434 myocardial ischaemia 415:434 myocardial ischaemia 415:434 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 4 33 theme site 692:695 arg1 architecture 697:708 active site architecture 685:708 active site architecture 685:708 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 3 34 theme CD73 511:514 arg1 form 557:560 a truncated form 545:560 a truncated form of CD73 that retains ecto-5'-nucleotidase activity 545:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 34 theme CD73 511:514 arg1 form 489:492 a soluble form 479:492 a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution 479:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 5 35 theme dimer 907:911 arg1 subunits 891:898 the two subunits 883:898 the two subunits of the dimer 883:911 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 3 36 theme crystal 458:464 arg1 structure 466:474 the crystal structure 454:474 the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity 454:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 0 37 theme form 31:34 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 0:84 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 5 38 attach linked 921:926 arg2 subunits 891:898 the two subunits 883:898 the two subunits of the dimer 883:911 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 5 38 attach linked 921:926 arg3 bridges 941:947 disulfide bridges 931:947 disulfide bridges 931:947 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 7 39 from roles 1293:1297 arg1 disease 1352:1358 disease 1352:1358 disease 1352:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 7 39 from roles 1293:1297 arg1 physiology 1337:1346 physiology 1337:1346 physiology 1337:1346 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 7 40 theme activator 1191:1199 arg1 molecules 1201:1209 activator molecules 1191:1209 activator molecules 1191:1209 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 3 41 theme truncated 547:555 arg1 form 557:560 a truncated form 545:560 a truncated form of CD73 that retains ecto-5'-nucleotidase activity 545:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 41 theme truncated 547:555 arg1 form 489:492 a soluble form 479:492 a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution 479:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 2 42 from metabolism 249:258 arg1 monophosphate 322:334 particular adenosine monophosphate 301:334 particular adenosine monophosphate 301:334 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 5 43 theme conserved 798:806 arg1 loop 808:811 a conserved loop 796:811 a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges 796:947 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 0 44 theme soluble 23:29 arg1 form 31:34 a soluble form 21:34 a soluble form of human CD73 21:48 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 7 45 from disease 1352:1358 arg1 roles 1293:1297 the possible roles 1280:1297 the possible roles of single nucleotide polymorphisms in physiology and disease 1280:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 3 46 theme ecto-5'-nucleotidase 583:602 arg1 activity 604:611 ecto-5'-nucleotidase activity 583:611 ecto-5'-nucleotidase activity 583:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 7 47 theme polymorphisms 1320:1332 arg1 roles 1293:1297 the possible roles 1280:1297 the possible roles of single nucleotide polymorphisms in physiology and disease 1280:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 2 48 contain has 193:195 arg1 CD73 188:191 CD73 188:191 CD73 188:191 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 2 48 contain has 193:195 arg2 functions 218:226 important regulatory functions 197:226 important regulatory functions 197:226 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 0 49 theme human 39:43 arg1 CD73 45:48 human CD73 39:48 human CD73 39:48 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 2 50 theme regulatory 207:216 arg1 functions 218:226 important regulatory functions 197:226 important regulatory functions 197:226 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 4 51 theme CD73 679:682 arg1 dimerisation 663:674 the dimerisation 659:674 the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein 659:763 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 2 52 theme important 197:205 arg1 functions 218:226 important regulatory functions 197:226 important regulatory functions 197:226 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 0 53 with structure 8:16 arg1 activity 76:83 ecto-5'-nucleotidase activity 55:83 ecto-5'-nucleotidase activity 55:83 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 3 54 theme soluble 481:487 arg1 form 557:560 a truncated form 545:560 a truncated form of CD73 that retains ecto-5'-nucleotidase activity 545:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 54 theme soluble 481:487 arg1 form 489:492 a soluble form 479:492 a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution 479:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 4 55 theme secondary 726:734 arg1 modifications 736:748 secondary modifications 726:748 secondary modifications of the protein 726:763 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 6 56 gly glycosylation 1011:1023 arg1 enzyme 1032:1037 the enzyme 1028:1037 the enzyme 1028:1037 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 5 57 theme disulfide 931:939 arg1 bridges 941:947 disulfide bridges 931:947 disulfide bridges 931:947 The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. 22997138 4 58 theme active 685:690 arg1 architecture 697:708 active site architecture 685:708 active site architecture 685:708 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 7 59 from physiology 1337:1346 arg1 roles 1293:1297 the possible roles 1280:1297 the possible roles of single nucleotide polymorphisms in physiology and disease 1280:1358 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 3 60 from resolution 533:542 arg1 sCD73 517:521 sCD73 517:521 sCD73 517:521 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 60 from resolution 533:542 arg1 CD73 511:514 human soluble CD73 497:514 human soluble CD73 (sCD73) at 2.2 Å resolution 497:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 60 from resolution 533:542 arg1 form 557:560 a truncated form 545:560 a truncated form of CD73 that retains ecto-5'-nucleotidase activity 545:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 60 from resolution 533:542 arg1 form 489:492 a soluble form 479:492 a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution 479:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 4 61 theme modifications 736:748 arg1 CD73 679:682 CD73 679:682 CD73 679:682 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 4 61 theme modifications 736:748 arg1 architecture 697:708 active site architecture 685:708 active site architecture 685:708 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 4 61 theme modifications 736:748 arg1 sense 717:721 a sense 715:721 a sense of secondary modifications of the protein 715:763 With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. 22997138 7 62 theme molecules 1201:1209 arg1 design 1167:1172 the design 1163:1172 the design of inhibitors or activator molecules 1163:1209 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 7 63 theme diseases 1240:1247 arg1 treatment 1219:1227 the treatment 1215:1227 the treatment of several diseases 1215:1247 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 2 64 theme extracellular 235:247 arg1 metabolism 249:258 the extracellular metabolism 231:258 the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, 231:335 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 1 65 theme membrane 178:185 arg1 side 159:162 the exterior side 146:162 the exterior side of the plasma membrane 146:185 CD73 is a dimeric ecto-5'-nucleotidase that is expressed on the exterior side of the plasma membrane. 22997138 3 66 theme human 497:501 arg1 sCD73 517:521 sCD73 517:521 sCD73 517:521 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 66 theme human 497:501 arg1 CD73 511:514 human soluble CD73 497:514 human soluble CD73 (sCD73) at 2.2 Å resolution 497:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 2 67 from monophosphates 282:295 arg1 monophosphate 322:334 particular adenosine monophosphate 301:334 particular adenosine monophosphate 301:334 CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. 22997138 6 68 theme microarray 968:977 arg1 imaging 979:985 biophotonic microarray imaging 956:985 biophotonic microarray imaging 956:985 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 22997138 0 69 theme CD73 45:48 arg1 form 31:34 a soluble form 21:34 a soluble form of human CD73 21:48 Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity. 22997138 3 70 theme CD73 565:568 arg1 form 557:560 a truncated form 545:560 a truncated form of CD73 that retains ecto-5'-nucleotidase activity 545:611 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 70 theme CD73 565:568 arg1 form 489:492 a soluble form 479:492 a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution 479:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 71 theme soluble 503:509 arg1 sCD73 517:521 sCD73 517:521 sCD73 517:521 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 3 71 theme soluble 503:509 arg1 CD73 511:514 human soluble CD73 497:514 human soluble CD73 (sCD73) at 2.2 Å resolution 497:542 Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. 22997138 7 72 theme crystal 1127:1133 arg1 structure 1135:1143 The crystal structure 1123:1143 The crystal structure of sCD73 1123:1152 The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease. 22997138 6 73 theme biophotonic 956:966 arg1 imaging 979:985 biophotonic microarray imaging 956:985 biophotonic microarray imaging 956:985 Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. 16107205 0 0 theme translation 75:85 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 0 theme translation 75:85 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 1 1 theme X 299:299 arg1 cDNAs 301:305 several mammalian DNase X cDNAs 275:305 several mammalian DNase X cDNAs 275:305 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 0 2 theme first 154:158 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 2 theme first 154:158 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 2 3 theme glycosylation 521:533 arg1 site 535:538 a potential N-linked glycosylation site 500:538 a potential N-linked glycosylation site 500:538 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 3 4 from Asn-243 725:731 arg1 glycosylation 708:720 an N-linked glycosylation 696:720 an N-linked glycosylation at Asn-243 696:731 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 2 5 theme C-terminal 546:555 arg1 domain 569:574 a C-terminal hydrophobic domain 544:574 a C-terminal hydrophobic domain 544:574 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 3 6 mod modified 684:691 arg1 X 589:589 Human DNase X 577:589 Human DNase X 577:589 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 6 mod modified 684:691 arg3 glycosylation 708:720 an N-linked glycosylation 696:720 an N-linked glycosylation at Asn-243 696:731 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 0 7 theme non-AUG 67:73 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 7 theme non-AUG 67:73 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 1 8 theme cDNAs 301:305 arg1 characterization 325:340 the molecular characterization 311:340 the molecular characterization of their coding proteins 311:365 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 1 8 theme cDNAs 301:305 arg1 isolation 262:270 the isolation 258:270 the isolation of several mammalian DNase X cDNAs 258:305 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 2 9 theme hydrophobic 557:567 arg1 domain 569:574 a C-terminal hydrophobic domain 544:574 a C-terminal hydrophobic domain 544:574 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 4 10 theme X 847:847 arg1 feature 824:830 a known feature 816:830 a known feature of human DNASE X 816:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 10 theme X 847:847 arg1 level 789:793 the high expression level 769:793 the high expression level in muscular tissues 769:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 0 11 theme DNase 166:170 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 11 theme DNase 166:170 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 5 12 theme conserved 1049:1057 arg1 triplet 1063:1069 a conserved CUG triplet 1047:1069 a conserved CUG triplet for translation initiation 1047:1096 We show that their mRNAs utilize a conserved CUG triplet for translation initiation. 16107205 0 13 theme human 160:164 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 13 theme human 160:164 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 5 14 theme CUG 1059:1061 arg1 triplet 1063:1069 a conserved CUG triplet 1047:1069 a conserved CUG triplet for translation initiation 1047:1096 We show that their mRNAs utilize a conserved CUG triplet for translation initiation. 16107205 4 15 theme human 835:839 arg1 X 847:847 human DNASE X 835:847 human DNASE X 835:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 1 16 theme present 232:238 arg1 study 240:244 the present study 228:244 the present study 228:244 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 2 17 theme signal 484:489 arg1 peptide 491:497 an N-terminal signal peptide 470:497 an N-terminal signal peptide 470:497 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 1 18 theme molecular 315:323 arg1 characterization 325:340 the molecular characterization 311:340 the molecular characterization of their coding proteins 311:365 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 2 19 theme N-linked 512:519 arg1 site 535:538 a potential N-linked glycosylation site 500:538 a potential N-linked glycosylation site 500:538 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 4 20 theme AUG 992:994 arg1 codon 1007:1011 an in-frame AUG initiation codon 980:1011 an in-frame AUG initiation codon 980:1011 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 3 21 gly glycosylation 708:720 arg2 Asn-243 725:731 Asn-243 725:731 Asn-243 725:731 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 21 gly glycosylation 708:720 arg1 Asn-243 725:731 Asn-243 725:731 Asn-243 725:731 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 0 22 theme porcine 101:107 arg1 mRNAs 120:124 porcine and bovine mRNAs 101:124 mRNAs 120:124 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 2 23 theme X 454:454 arg1 proteins 456:463 all the mammalian DNase X proteins 430:463 all the mammalian DNase X proteins 430:463 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 2 24 theme DNase 448:452 arg1 proteins 456:463 all the mammalian DNase X proteins 430:463 all the mammalian DNase X proteins 430:463 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 4 25 theme bovine 931:936 arg1 proteins 946:953 bovine DNase X proteins 931:953 bovine DNase X proteins 931:953 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 3 26 theme N-linked 699:706 arg1 glycosylation 708:720 an N-linked glycosylation 696:720 an N-linked glycosylation at Asn-243 696:731 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 27 located located 635:641 arg1 ER 650:651 the ER 646:651 the ER (endoplasmic reticulum) 646:675 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 27 located located 635:641 arg1 reticulum 666:674 endoplasmic reticulum 654:674 endoplasmic reticulum 654:674 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 27 located located 635:641 arg2 X 589:589 Human DNase X 577:589 Human DNase X 577:589 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 0 28 with homologous 200:209 arg1 DNase 216:220 DNase I 216:222 DNase I 216:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 4 29 theme muscular 798:805 arg1 tissues 807:813 muscular tissues 798:813 muscular tissues 798:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 2 30 link N-linked 512:519 arg1 site 535:538 a potential N-linked glycosylation site 500:538 a potential N-linked glycosylation site 500:538 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 0 31 theme Physical 0:7 arg1 properties 25:34 Physical and biochemical properties 0:34 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 0:223 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 4 32 theme codon 1007:1011 arg1 absence 969:975 the absence 965:975 the absence of an in-frame AUG initiation codon 965:1011 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 3 33 theme S3 622:623 arg1 cells 625:629 HeLa S3 cells 617:629 HeLa S3 cells 617:629 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 2 34 theme potential 502:510 arg1 site 535:538 a potential N-linked glycosylation site 500:538 a potential N-linked glycosylation site 500:538 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 0 35 theme mRNAs 120:124 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 35 theme mRNAs 120:124 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 4 36 theme initiation 996:1005 arg1 codon 1007:1011 an in-frame AUG initiation codon 980:1011 an in-frame AUG initiation codon 980:1011 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 0 37 theme biochemical 13:23 arg1 properties 25:34 Physical and biochemical properties 0:34 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 0:223 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 5 38 theme translation 1075:1085 arg1 initiation 1087:1096 translation initiation 1075:1096 translation initiation 1075:1096 We show that their mRNAs utilize a conserved CUG triplet for translation initiation. 16107205 0 39 theme bovine 113:118 arg1 mRNAs 120:124 porcine and bovine mRNAs 101:124 mRNAs 120:124 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 3 40 theme HeLa 617:620 arg1 cells 625:629 HeLa S3 cells 617:629 HeLa S3 cells 617:629 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 4 41 from level 789:793 arg1 tissues 807:813 muscular tissues 798:813 muscular tissues 798:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 0 42 theme mammalian 39:47 arg1 proteins 57:64 mammalian DNase X proteins 39:64 mammalian DNase X proteins 39:64 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 43 theme DNase 130:134 arg1 X 145:145 DNase X. DNase X 130:145 DNase X. DNase X 130:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 1 44 theme coding 351:356 arg1 proteins 358:365 their coding proteins 345:365 their coding proteins 345:365 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 4 45 theme expression 739:748 arg1 analyses 750:757 Gene expression analyses 734:757 Gene expression analyses 734:757 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 46 theme proteins 946:953 arg1 translation 904:914 the translation 900:914 the translation of porcine and bovine DNase X proteins 900:953 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 2 47 theme sequence 370:377 arg1 comparison 379:388 A sequence comparison 368:388 A sequence comparison 368:388 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 2 48 theme mammalian 438:446 arg1 proteins 456:463 all the mammalian DNase X proteins 430:463 all the mammalian DNase X proteins 430:463 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 4 49 theme porcine 919:925 arg1 translation 904:914 the translation 900:914 the translation of porcine and bovine DNase X proteins 900:953 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 1 50 theme proteins 358:365 arg1 characterization 325:340 the molecular characterization 311:340 the molecular characterization of their coding proteins 311:365 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 1 50 theme proteins 358:365 arg1 isolation 262:270 the isolation 258:270 the isolation of several mammalian DNase X cDNAs 258:305 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 2 51 theme conserved 403:411 arg1 characteristics 413:427 some conserved characteristics 398:427 some conserved characteristics 398:427 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 3 52 link N-linked 699:706 arg1 glycosylation 708:720 an N-linked glycosylation 696:720 an N-linked glycosylation at Asn-243 696:731 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 2 53 theme N-terminal 473:482 arg1 peptide 491:497 an N-terminal signal peptide 470:497 an N-terminal signal peptide 470:497 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 0 54 theme X 55:55 arg1 proteins 57:64 mammalian DNase X proteins 39:64 mammalian DNase X proteins 39:64 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 55 theme DNase 139:143 arg1 X 145:145 DNase X. DNase X 130:145 DNase X. DNase X 130:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 4 56 theme high 773:776 arg1 feature 824:830 a known feature 816:830 a known feature of human DNASE X 816:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 56 theme high 773:776 arg1 level 789:793 the high expression level 769:793 the high expression level in muscular tissues 769:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 3 57 theme endoplasmic 654:664 arg1 ER 650:651 the ER 646:651 the ER (endoplasmic reticulum) 646:675 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 57 theme endoplasmic 654:664 arg1 reticulum 666:674 endoplasmic reticulum 654:674 endoplasmic reticulum 654:674 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 0 58 theme DNase 49:53 arg1 proteins 57:64 mammalian DNase X proteins 39:64 mammalian DNase X proteins 39:64 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 1 59 theme several 275:281 arg1 cDNAs 301:305 several mammalian DNase X cDNAs 275:305 several mammalian DNase X cDNAs 275:305 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 0 60 theme X. 136:137 arg1 X 145:145 DNase X. DNase X 130:145 DNase X. DNase X 130:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 4 61 theme expression 778:787 arg1 feature 824:830 a known feature 816:830 a known feature of human DNASE X 816:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 61 theme expression 778:787 arg1 level 789:793 the high expression level 769:793 the high expression level in muscular tissues 769:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 2 62 contain have 465:468 arg2 peptide 491:497 an N-terminal signal peptide 470:497 an N-terminal signal peptide 470:497 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 2 62 contain have 465:468 arg1 proteins 456:463 all the mammalian DNase X proteins 430:463 all the mammalian DNase X proteins 430:463 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 2 62 contain have 465:468 arg2 domain 569:574 a C-terminal hydrophobic domain 544:574 a C-terminal hydrophobic domain 544:574 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 2 62 contain have 465:468 arg2 site 535:538 a potential N-linked glycosylation site 500:538 a potential N-linked glycosylation site 500:538 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 16107205 4 63 theme known 818:822 arg1 feature 824:830 a known feature 816:830 a known feature of human DNASE X 816:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 63 theme known 818:822 arg1 level 789:793 the high expression level 769:793 the high expression level in muscular tissues 769:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 1 64 theme mammalian 283:291 arg1 cDNAs 301:305 several mammalian DNase X cDNAs 275:305 several mammalian DNase X cDNAs 275:305 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 4 65 theme DNASE 841:845 arg1 X 847:847 human DNASE X 835:847 human DNASE X 835:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 66 theme Gene 734:737 arg1 analyses 750:757 Gene expression analyses 734:757 Gene expression analyses 734:757 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 0 67 dep properties 25:34 arg1 initiation 87:96 non-AUG translation initiation 67:96 non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X 67:145 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 0 67 dep properties 25:34 arg1 protein 172:178 the first human DNase protein 150:178 the first human DNase protein identified as being homologous with DNase I 150:222 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 3 68 theme Human 577:581 arg1 X 589:589 Human DNase X 577:589 Human DNase X 577:589 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 69 from ER 650:651 arg1 located 635:641 located 635:641 located 635:641 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 4 70 theme DNase 938:942 arg1 proteins 946:953 bovine DNase X proteins 931:953 bovine DNase X proteins 931:953 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 71 located observed 858:865 arg2 feature 824:830 a known feature 816:830 a known feature of human DNASE X 816:847 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 71 located observed 858:865 arg2 level 789:793 the high expression level 769:793 the high expression level in muscular tissues 769:813 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 4 71 located observed 858:865 arg1 X. 882:883 X. 882:883 X. 882:883 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 3 72 from located 635:641 arg1 ER 650:651 the ER 646:651 the ER (endoplasmic reticulum) 646:675 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 72 from located 635:641 arg1 reticulum 666:674 endoplasmic reticulum 654:674 endoplasmic reticulum 654:674 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 3 73 theme DNase 583:587 arg1 X 589:589 Human DNase X 577:589 Human DNase X 577:589 Human DNase X, ectopically expressed in HeLa S3 cells, is located in the ER (endoplasmic reticulum) and is modified by an N-linked glycosylation at Asn-243. 16107205 4 74 theme in-frame 983:990 arg1 codon 1007:1011 an in-frame AUG initiation codon 980:1011 an in-frame AUG initiation codon 980:1011 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 0 75 theme proteins 57:64 arg1 properties 25:34 Physical and biochemical properties 0:34 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 0:223 Physical and biochemical properties of mammalian DNase X proteins: non-AUG translation initiation of porcine and bovine mRNAs for DNase X. DNase X is the first human DNase protein identified as being homologous with DNase I. 16107205 1 76 theme DNase 293:297 arg1 cDNAs 301:305 several mammalian DNase X cDNAs 275:305 several mammalian DNase X cDNAs 275:305 In the present study we describe the isolation of several mammalian DNase X cDNAs and the molecular characterization of their coding proteins. 16107205 4 77 theme X 944:944 arg1 proteins 946:953 bovine DNase X proteins 931:953 bovine DNase X proteins 931:953 Gene expression analyses show that the high expression level in muscular tissues, a known feature of human DNASE X, is also observed in mouse DNase X. Interestingly, the translation of porcine and bovine DNase X proteins occurs in the absence of an in-frame AUG initiation codon. 16107205 2 78 gly glycosylation 521:533 arg2 site 535:538 a potential N-linked glycosylation site 500:538 a potential N-linked glycosylation site 500:538 A sequence comparison reveals some conserved characteristics: all the mammalian DNase X proteins have an N-terminal signal peptide, a potential N-linked glycosylation site and a C-terminal hydrophobic domain. 22235133 8 0 theme V-shaped 1203:1210 arg1 receptor 1212:1219 the V-shaped receptor 1199:1219 the V-shaped receptor 1199:1219 The glycan thus transforms the V-shaped receptor into an A-frame. 22235133 9 1 theme structural 1266:1275 arg1 explanation 1277:1287 a novel structural explanation 1258:1287 a novel structural explanation of the role of the class I cytokine signature motif 1258:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 3 2 theme receptor 444:451 arg1 hallmark 377:384 A hallmark 375:384 A hallmark of the class I cytokine receptors 375:418 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 3 2 theme receptor 444:451 arg1 motif 463:467 the class I cytokine receptor signature motif 423:467 the class I cytokine receptor signature motif (WSXWS) 423:475 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 6 3 theme WSXWS 871:875 arg1 C-mannosylated 896:909 C-mannosylated 896:909 C-mannosylated 896:909 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 6 3 theme WSXWS 871:875 arg1 motif 877:881 the WSXWS motif 867:881 the WSXWS motif of IL-21R 867:891 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 6 4 theme first 918:922 arg1 tryptophan 924:933 the first tryptophan 914:933 the first tryptophan 914:933 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 3 5 theme signature 453:461 arg1 hallmark 377:384 A hallmark 375:384 A hallmark of the class I cytokine receptors 375:418 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 3 5 theme signature 453:461 arg1 motif 463:467 the class I cytokine receptor signature motif 423:467 the class I cytokine receptor signature motif (WSXWS) 423:475 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 1 6 theme innate 219:224 arg1 responses 246:254 both innate and adaptive immune responses 214:254 both innate and adaptive immune responses 214:254 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 0 7 theme sugar 82:86 arg1 chain 88:92 sugar chain 82:92 sugar chain interacting with WSXWS motif 82:121 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 6 8 from tryptophan 924:933 arg1 C-mannosylated 896:909 C-mannosylated 896:909 C-mannosylated 896:909 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 6 8 from tryptophan 924:933 arg1 motif 877:881 the WSXWS motif 867:881 the WSXWS motif of IL-21R 867:891 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 7 9 theme extensive 1111:1119 arg1 network 1138:1144 an extensive hydrogen bonding network 1108:1144 an extensive hydrogen bonding network 1108:1144 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 2 10 theme IL-21 327:331 arg1 IL-21R 343:348 IL-21R 343:348 IL-21R 343:348 It signals through a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain. 22235133 2 10 theme IL-21 327:331 arg1 receptor 333:340 IL-21 receptor 327:340 the IL-21 receptor (IL-21R) 323:349 It signals through a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain. 22235133 9 11 theme motif 1335:1339 arg1 role 1296:1299 the role 1292:1299 the role of the class I cytokine signature motif 1292:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 3 12 theme class 427:431 arg1 receptor 444:451 class I cytokine receptor 427:451 the class I cytokine receptor signature motif (WSXWS) 423:475 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 7 13 theme IL-21R 1062:1067 arg1 IL-21R 1062:1067 IL-21R 1062:1067 IL-21R 1062:1067 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 7 13 theme IL-21R 1062:1067 arg1 domain 1052:1057 the extracellular domain 1034:1057 the extracellular domain of IL-21R 1034:1067 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 4 14 theme receptor 616:623 arg1 internalization 625:639 receptor internalization 616:639 receptor internalization 616:639 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 9 15 theme cytokine 1316:1323 arg1 motif 1335:1339 the class I cytokine signature motif 1304:1339 the class I cytokine signature motif 1304:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 9 16 theme class 1308:1312 arg1 motif 1335:1339 the class I cytokine signature motif 1304:1339 the class I cytokine signature motif 1304:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 1 17 theme adaptive 230:237 arg1 responses 246:254 both innate and adaptive immune responses 214:254 both innate and adaptive immune responses 214:254 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 7 18 theme sugar 970:974 arg1 chain 976:980 a sugar chain 968:980 a sugar chain 968:980 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 6 19 theme IL-21R 886:891 arg1 C-mannosylated 896:909 C-mannosylated 896:909 C-mannosylated 896:909 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 6 19 theme IL-21R 886:891 arg1 motif 877:881 the WSXWS motif 867:881 the WSXWS motif of IL-21R 867:891 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 3 20 theme class 393:397 arg1 receptors 410:418 the class I cytokine receptors 389:418 the class I cytokine receptors 389:418 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 7 21 theme WSXWS 1088:1092 arg1 motif 1094:1098 the WSXWS motif 1084:1098 the WSXWS motif 1084:1098 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 2 22 theme heterodimeric 278:290 arg1 complex 301:307 a heterodimeric receptor complex 276:307 a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain 276:372 It signals through a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain. 22235133 5 23 theme consensus 747:755 arg1 motif 724:728 the WXXW motif 715:728 the WXXW motif 715:728 Furthermore, the WXXW motif is known to be a consensus sequence for C-mannosylation. 22235133 5 23 theme consensus 747:755 arg1 sequence 757:764 a consensus sequence 745:764 a consensus sequence for C-mannosylation 745:784 Furthermore, the WXXW motif is known to be a consensus sequence for C-mannosylation. 22235133 0 24 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 0:67 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 1 25 theme immune 239:244 arg1 responses 246:254 both innate and adaptive immune responses 214:254 both innate and adaptive immune responses 214:254 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 4 26 theme proper 642:647 arg1 folding 649:655 proper folding 642:655 proper folding 642:655 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 4 27 theme diverse 571:577 arg1 transduction 688:699 signal transduction 681:699 signal transduction 681:699 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 4 27 theme diverse 571:577 arg1 internalization 625:639 receptor internalization 616:639 receptor internalization 616:639 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 4 27 theme diverse 571:577 arg1 export 662:667 export 662:667 export 662:667 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 4 27 theme diverse 571:577 arg1 binding 607:613 ligand binding 600:613 ligand binding 600:613 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 4 27 theme diverse 571:577 arg1 folding 649:655 proper folding 642:655 proper folding 642:655 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 4 27 theme diverse 571:577 arg1 functions 579:587 diverse functions 571:587 diverse functions 571:587 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 0 28 theme interleukin-21 21:34 arg1 IL-21R 46:51 IL-21R 46:51 IL-21R 46:51 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 0 28 theme interleukin-21 21:34 arg1 receptor 36:43 interleukin-21 receptor 21:43 interleukin-21 receptor (IL-21R) 21:52 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 4 29 theme exact 482:486 arg1 role 488:491 The exact role 478:491 The exact role of this motif 478:505 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 0 30 theme WSXWS 111:115 arg1 motif 117:121 WSXWS motif 111:121 WSXWS motif 111:121 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 3 31 dep motif 463:467 arg1 WSXWS 470:474 WSXWS 470:474 WSXWS 470:474 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 1 32 theme class 162:166 arg1 cytokine 170:177 a class I cytokine 160:177 a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses 160:254 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 1 32 theme class 162:166 arg1 IL-21 151:155 IL-21 151:155 IL-21 151:155 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 5 33 theme WXXW 719:722 arg1 motif 724:728 the WXXW motif 715:728 the WXXW motif 715:728 Furthermore, the WXXW motif is known to be a consensus sequence for C-mannosylation. 22235133 5 33 theme WXXW 719:722 arg1 sequence 757:764 a consensus sequence 745:764 a consensus sequence for C-mannosylation 745:784 Furthermore, the WXXW motif is known to be a consensus sequence for C-mannosylation. 22235133 4 34 theme ligand 600:605 arg1 binding 607:613 ligand binding 600:613 ligand binding 600:613 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 1 35 theme I 168:168 arg1 cytokine 170:177 a class I cytokine 160:177 a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses 160:254 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 1 35 theme I 168:168 arg1 IL-21 151:155 IL-21 151:155 IL-21 151:155 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 6 36 gly C-mannosylated 896:909 arg1 C-mannosylated 896:909 C-mannosylated 896:909 C-mannosylated 896:909 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 6 36 gly C-mannosylated 896:909 arg2 tryptophan 924:933 the first tryptophan 914:933 the first tryptophan 914:933 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 6 36 gly C-mannosylated 896:909 arg1 motif 877:881 the WSXWS motif 867:881 the WSXWS motif of IL-21R 867:891 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 9 37 theme signature 1325:1333 arg1 motif 1335:1339 the class I cytokine signature motif 1304:1339 the class I cytokine signature motif 1304:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 0 38 theme receptor 36:43 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 0:67 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 6 39 theme IL-21 829:833 arg1 structure 816:824 the crystal structure 804:824 the crystal structure of IL-21 bound to IL-21R 804:849 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 4 40 theme motif 501:505 arg1 role 488:491 The exact role 478:491 The exact role of this motif 478:505 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 2 41 theme common 359:364 arg1 γ-chain 366:372 the common γ-chain 355:372 the common γ-chain 355:372 It signals through a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain. 22235133 7 42 theme fibronectin 998:1008 arg1 domains 1010:1016 the two fibronectin domains 990:1016 the two fibronectin domains that constitute the extracellular domain of IL-21R 990:1067 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 3 43 theme I 433:433 arg1 receptor 444:451 class I cytokine receptor 427:451 the class I cytokine receptor signature motif (WSXWS) 423:475 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 0 44 theme integral 126:133 arg1 part 135:138 integral part 126:138 integral part of IL-21R 126:148 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 3 45 theme cytokine 435:442 arg1 receptor 444:451 class I cytokine receptor 427:451 the class I cytokine receptor signature motif (WSXWS) 423:475 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 6 46 theme crystal 808:814 arg1 structure 816:824 the crystal structure 804:824 the crystal structure of IL-21 bound to IL-21R 804:849 Here, we present the crystal structure of IL-21 bound to IL-21R and reveal that the WSXWS motif of IL-21R is C-mannosylated at the first tryptophan. 22235133 0 47 theme IL-21R 143:148 arg1 part 135:138 integral part 126:138 integral part of IL-21R 126:148 Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. 22235133 4 48 theme signal 681:686 arg1 transduction 688:699 signal transduction 681:699 signal transduction 681:699 The exact role of this motif has not been determined yet; however, it has been implicated in diverse functions, including ligand binding, receptor internalization, proper folding, and export, as well as signal transduction. 22235133 9 49 theme novel 1260:1264 arg1 explanation 1277:1287 a novel structural explanation 1258:1287 a novel structural explanation of the role of the class I cytokine signature motif 1258:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 9 50 theme role 1296:1299 arg1 explanation 1277:1287 a novel structural explanation 1258:1287 a novel structural explanation of the role of the class I cytokine signature motif 1258:1339 This finding offers a novel structural explanation of the role of the class I cytokine signature motif. 22235133 2 51 theme receptor 292:299 arg1 complex 301:307 a heterodimeric receptor complex 276:307 a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain 276:372 It signals through a heterodimeric receptor complex consisting of the IL-21 receptor (IL-21R) and the common γ-chain. 22235133 1 52 theme pleiotropic 191:201 arg1 effects 203:209 pleiotropic effects 191:209 pleiotropic effects 191:209 IL-21 is a class I cytokine that exerts pleiotropic effects on both innate and adaptive immune responses. 22235133 3 53 theme cytokine 401:408 arg1 receptors 410:418 the class I cytokine receptors 389:418 the class I cytokine receptors 389:418 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 3 54 theme receptors 410:418 arg1 hallmark 377:384 A hallmark 375:384 A hallmark of the class I cytokine receptors 375:418 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 3 54 theme receptors 410:418 arg1 motif 463:467 the class I cytokine receptor signature motif 423:467 the class I cytokine receptor signature motif (WSXWS) 423:475 A hallmark of the class I cytokine receptors is the class I cytokine receptor signature motif (WSXWS). 22235133 7 55 theme hydrogen 1121:1128 arg1 network 1138:1144 an extensive hydrogen bonding network 1108:1144 an extensive hydrogen bonding network 1108:1144 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 7 56 theme extracellular 1038:1050 arg1 IL-21R 1062:1067 IL-21R 1062:1067 IL-21R 1062:1067 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 7 56 theme extracellular 1038:1050 arg1 domain 1052:1057 the extracellular domain 1034:1057 the extracellular domain of IL-21R 1034:1067 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 22235133 7 57 theme bonding 1130:1136 arg1 network 1138:1144 an extensive hydrogen bonding network 1108:1144 an extensive hydrogen bonding network 1108:1144 We furthermore demonstrate that a sugar chain bridges the two fibronectin domains that constitute the extracellular domain of IL-21R and anchors at the WSXWS motif through an extensive hydrogen bonding network, including mannosylation. 8489250 1 0 gly glycoprotein 235:246 arg1 factor 213:218 recombinant human differentiation-stimulating factor 167:218 recombinant human differentiation-stimulating factor 167:218 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 0 gly glycoprotein 235:246 arg1 glycoprotein 235:246 a 180-residue glycoprotein 221:246 a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 221:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 3 1 dep Cys-131 606:612 arg1 to 603:604 to 603:604 to 603:604 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 2 theme disulfide 579:587 arg1 Cys-134 625:631 Cys-12 to Cys-134 615:631 Cys-12 to Cys-134 615:631 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 2 theme disulfide 579:587 arg1 bonds 589:593 three intrachain disulfide bonds 562:593 three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163) 562:655 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 2 theme disulfide 579:587 arg1 Cys-60 638:643 Cys-60 638:643 Cys-60 638:643 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 2 theme disulfide 579:587 arg1 Cys-131 606:612 Cys-18 to Cys-131 596:612 Cys-18 to Cys-131 596:612 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 3 theme Cys-18 596:601 arg1 bonds 589:593 three intrachain disulfide bonds 562:593 three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163) 562:655 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 3 theme Cys-18 596:601 arg1 Cys-131 606:612 Cys-18 to Cys-131 596:612 Cys-18 to Cys-131 596:612 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 2 4 contain containing 328:337 arg1 peptides 319:326 Peptide peptides 311:326 Peptide peptides containing six potential N-glycosylation sites 311:373 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 2 4 contain containing 328:337 arg2 sites 369:373 six potential N-glycosylation sites 339:373 six potential N-glycosylation sites 339:373 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 3 5 theme bonds 589:593 arg1 assignments 547:557 the assignments 543:557 the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163) 543:655 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 2 6 theme Asn 407:409 arg1 residues 411:418 Asn residues 9, 34, 63, 73, 96, and 116 407:445 Asn residues 9, 34, 63, 73, 96, and 116 407:445 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 2 7 theme N-glycosylation 353:367 arg1 sites 369:373 six potential N-glycosylation sites 339:373 six potential N-glycosylation sites 339:373 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 1 8 theme 180-residue 223:233 arg1 factor 213:218 recombinant human differentiation-stimulating factor 167:218 recombinant human differentiation-stimulating factor 167:218 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 8 theme 180-residue 223:233 arg1 glycoprotein 235:246 a 180-residue glycoprotein 221:246 a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 221:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 2 9 gly N-glycosylation 353:367 arg2 six 339:341 six 339:341 six 339:341 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 2 9 gly N-glycosylation 353:367 arg2 sites 369:373 six potential N-glycosylation sites 339:373 six potential N-glycosylation sites 339:373 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 2 10 theme potential 343:351 arg1 sites 369:373 six potential N-glycosylation sites 339:373 six potential N-glycosylation sites 339:373 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 0 11 theme Glycosylation 0:12 arg1 pattern 14:20 Glycosylation pattern 0:20 Glycosylation pattern 0:20 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 3 12 dep bonds 589:593 arg1 bonds 589:593 three intrachain disulfide bonds 562:593 three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163) 562:655 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 12 dep bonds 589:593 arg1 Cys-134 625:631 Cys-12 to Cys-134 615:631 Cys-12 to Cys-134 615:631 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 12 dep bonds 589:593 arg1 Cys-60 638:643 Cys-60 638:643 Cys-60 638:643 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 12 dep bonds 589:593 arg1 Cys-131 606:612 Cys-18 to Cys-131 596:612 Cys-18 to Cys-131 596:612 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 13 theme Additional 462:471 arg1 peptides 473:480 Additional peptides 462:480 Additional peptides 462:480 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 0 14 theme disulfide 26:34 arg1 assignments 36:46 disulfide assignments 26:46 disulfide assignments 26:46 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 3 15 theme tryptic 496:502 arg1 digestion 504:512 tryptic digestion 496:512 tryptic digestion of peptic fragments 496:532 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 1 16 theme post-translational 131:148 arg1 modifications 150:162 the post-translational modifications 127:162 the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 127:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 3 17 dep Cys-134 625:631 arg1 to 622:623 to 622:623 to 622:623 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 1 18 theme transfected 270:280 arg1 cells 304:308 transfected Chinese hamster ovary cells 270:308 transfected Chinese hamster ovary cells 270:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 19 theme Chinese 282:288 arg1 cells 304:308 transfected Chinese hamster ovary cells 270:308 transfected Chinese hamster ovary cells 270:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 0 20 theme human 63:67 arg1 factor 97:102 recombinant human differentiation-stimulating factor 51:102 recombinant human differentiation-stimulating factor 51:102 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 3 21 theme fragments 524:532 arg1 digestion 504:512 tryptic digestion 496:512 tryptic digestion of peptic fragments 496:532 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 0 22 theme recombinant 51:61 arg1 factor 97:102 recombinant human differentiation-stimulating factor 51:102 recombinant human differentiation-stimulating factor 51:102 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 3 23 theme Cys-12 615:620 arg1 bonds 589:593 three intrachain disulfide bonds 562:593 three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163) 562:655 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 23 theme Cys-12 615:620 arg1 Cys-134 625:631 Cys-12 to Cys-134 615:631 Cys-12 to Cys-134 615:631 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 2 24 used utilized 452:459 arg2 residues 411:418 Asn residues 9, 34, 63, 73, 96, and 116 407:445 Asn residues 9, 34, 63, 73, 96, and 116 407:445 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 1 25 theme recombinant 167:177 arg1 factor 213:218 recombinant human differentiation-stimulating factor 167:218 recombinant human differentiation-stimulating factor 167:218 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 25 theme recombinant 167:177 arg1 glycoprotein 235:246 a 180-residue glycoprotein 221:246 a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 221:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 26 theme hamster 290:296 arg1 cells 304:308 transfected Chinese hamster ovary cells 270:308 transfected Chinese hamster ovary cells 270:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 3 27 theme intrachain 568:577 arg1 Cys-134 625:631 Cys-12 to Cys-134 615:631 Cys-12 to Cys-134 615:631 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 27 theme intrachain 568:577 arg1 bonds 589:593 three intrachain disulfide bonds 562:593 three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163) 562:655 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 27 theme intrachain 568:577 arg1 Cys-60 638:643 Cys-60 638:643 Cys-60 638:643 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 3 27 theme intrachain 568:577 arg1 Cys-131 606:612 Cys-18 to Cys-131 596:612 Cys-18 to Cys-131 596:612 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 1 28 theme human 179:183 arg1 factor 213:218 recombinant human differentiation-stimulating factor 167:218 recombinant human differentiation-stimulating factor 167:218 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 28 theme human 179:183 arg1 glycoprotein 235:246 a 180-residue glycoprotein 221:246 a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 221:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 29 theme ovary 298:302 arg1 cells 304:308 transfected Chinese hamster ovary cells 270:308 transfected Chinese hamster ovary cells 270:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 0 30 theme factor 97:102 arg1 pattern 14:20 Glycosylation pattern 0:20 Glycosylation pattern 0:20 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 0 30 theme factor 97:102 arg1 assignments 36:46 disulfide assignments 26:46 disulfide assignments 26:46 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 1 31 theme differentiation-stimulating 185:211 arg1 factor 213:218 recombinant human differentiation-stimulating factor 167:218 recombinant human differentiation-stimulating factor 167:218 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 1 31 theme differentiation-stimulating 185:211 arg1 glycoprotein 235:246 a 180-residue glycoprotein 221:246 a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 221:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 8489250 3 32 theme peptic 517:522 arg1 fragments 524:532 peptic fragments 517:532 peptic fragments 517:532 Additional peptides, generated by tryptic digestion of peptic fragments, allowed the assignments of three intrachain disulfide bonds (Cys-18 to Cys-131, Cys-12 to Cys-134, and Cys-60 to Cys-163). 8489250 0 33 theme differentiation-stimulating 69:95 arg1 factor 97:102 recombinant human differentiation-stimulating factor 51:102 recombinant human differentiation-stimulating factor 51:102 Glycosylation pattern and disulfide assignments of recombinant human differentiation-stimulating factor. 8489250 2 34 theme Peptide 311:317 arg1 peptides 319:326 Peptide peptides 311:326 Peptide peptides containing six potential N-glycosylation sites 311:373 Peptide peptides containing six potential N-glycosylation sites were analyzed to determine that Asn residues 9, 34, 63, 73, 96, and 116 were utilized. 8489250 1 35 theme factor 213:218 arg1 modifications 150:162 the post-translational modifications 127:162 the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells 127:308 This report describes the post-translational modifications of recombinant human differentiation-stimulating factor, a 180-residue glycoprotein that is secreted from transfected Chinese hamster ovary cells. 10644446 4 0 theme angiopoietins 508:520 arg1 structure 495:503 the characteristic structure 476:503 the characteristic structure of angiopoietins 476:520 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 6 1 theme angiopoietins 997:1009 arg1 FHD 990:992 the FHD 986:992 the FHD of angiopoietins 1, 2, and 4 986:1021 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 4 2 theme dimeric 586:592 arg1 coiled-coils 606:617 dimeric or trimeric coiled-coils 586:617 dimeric or trimeric coiled-coils 586:617 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 3 theme signal 525:530 arg1 peptide 532:538 a signal peptide 523:538 a signal peptide 523:538 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 4 dep polypeptide 459:469 arg1 FHD 687:689 FHD 687:689 FHD 687:689 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 4 dep polypeptide 459:469 arg1 domain 561:566 an extended helical domain 541:566 an extended helical domain predicted to form dimeric or trimeric coiled-coils 541:617 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 4 dep polypeptide 459:469 arg1 peptide 532:538 a signal peptide 523:538 a signal peptide 523:538 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 4 dep polypeptide 459:469 arg1 peptide 635:641 a short linker peptide 620:641 a short linker peptide 620:641 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 4 dep polypeptide 459:469 arg1 domain 679:684 a globular fibrinogen homology domain 648:684 a globular fibrinogen homology domain (FHD) 648:690 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 3 5 theme liver 361:365 arg1 development 367:377 liver development 361:377 liver development 361:377 ANGPTL3 is expressed early during liver development, and expression is maintained in adult liver. 10644446 6 6 theme intramolecular 873:886 arg1 bonds 898:902 the intramolecular disulfide bonds 869:902 the intramolecular disulfide bonds within the FHD 869:917 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 6 7 located found 973:977 arg1 FHD 990:992 the FHD 986:992 the FHD of angiopoietins 1, 2, and 4 986:1021 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 6 7 located found 973:977 arg2 cysteines 954:962 two other cysteines 944:962 two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4 944:1021 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 4 8 theme trimeric 597:604 arg1 coiled-coils 606:617 dimeric or trimeric coiled-coils 586:617 dimeric or trimeric coiled-coils 586:617 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 5 9 theme 455-acid 713:720 arg1 polypeptide 722:732 a 455-acid polypeptide 711:732 a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA 711:806 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 5 9 theme 455-acid 713:720 arg1 ANGPTL3 700:706 Murine ANGPTL3 693:706 Murine ANGPTL3 693:706 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 4 10 theme characteristic 480:493 arg1 structure 495:503 the characteristic structure 476:503 the characteristic structure of angiopoietins 476:520 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 7 11 theme binding 1081:1087 arg1 motif 1089:1093 the characteristic calcium binding motif 1054:1093 the characteristic calcium binding motif found in the other angiopoietins 1054:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 6 12 contain contains 817:824 arg2 cysteines 845:853 the four conserved cysteines 826:853 the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD 826:917 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 6 12 contain contains 817:824 arg1 ANGPTL3 809:815 ANGPTL3 809:815 ANGPTL3 809:815 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 5 13 theme DNA 804:806 arg1 kb 798:799 about 11 kb 789:799 about 11 kb of DNA 789:806 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 8 14 theme genes 1184:1188 arg1 mapping 1149:1155 radiation hybrid mapping 1132:1155 radiation hybrid mapping 1132:1155 By radiation hybrid mapping and the use of surrounding genes, human ANGPTL3 maps to the 1p31 region. 10644446 8 14 theme genes 1184:1188 arg1 use 1165:1167 the use 1161:1167 the use of surrounding genes 1161:1188 By radiation hybrid mapping and the use of surrounding genes, human ANGPTL3 maps to the 1p31 region. 10644446 4 15 theme globular 650:657 arg1 FHD 687:689 FHD 687:689 FHD 687:689 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 15 theme globular 650:657 arg1 domain 679:684 a globular fibrinogen homology domain 648:684 a globular fibrinogen homology domain (FHD) 648:690 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 0 16 theme protein 51:57 arg1 Identification 0:13 Identification 0:13 Identification of a mammalian angiopoietin-related protein 0:57 Identification of a mammalian angiopoietin-related protein expressed specifically in liver. 10644446 7 17 theme other 1108:1112 arg1 angiopoietins 1114:1126 the other angiopoietins 1104:1126 the other angiopoietins 1104:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 4 18 with polypeptide 459:469 arg1 structure 495:503 the characteristic structure 476:503 the characteristic structure of angiopoietins 476:520 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 19 theme homology 670:677 arg1 FHD 687:689 FHD 687:689 FHD 687:689 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 19 theme homology 670:677 arg1 domain 679:684 a globular fibrinogen homology domain 648:684 a globular fibrinogen homology domain (FHD) 648:690 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 6 20 theme disulfide 888:896 arg1 bonds 898:902 the intramolecular disulfide bonds 869:902 the intramolecular disulfide bonds within the FHD 869:917 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 5 21 from exons 751:755 arg1 chromosome 766:775 mouse chromosome 4 760:777 mouse chromosome 4 760:777 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 4 22 theme fibrinogen 659:668 arg1 FHD 687:689 FHD 687:689 FHD 687:689 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 22 theme fibrinogen 659:668 arg1 domain 679:684 a globular fibrinogen homology domain 648:684 a globular fibrinogen homology domain (FHD) 648:690 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 0 23 theme mammalian 20:28 arg1 protein 51:57 a mammalian angiopoietin-related protein 18:57 a mammalian angiopoietin-related protein 18:57 Identification of a mammalian angiopoietin-related protein expressed specifically in liver. 10644446 4 24 theme helical 553:559 arg1 domain 561:566 an extended helical domain 541:566 an extended helical domain predicted to form dimeric or trimeric coiled-coils 541:617 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 7 25 located found 1095:1099 arg1 angiopoietins 1114:1126 the other angiopoietins 1104:1126 the other angiopoietins 1104:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 7 25 located found 1095:1099 arg2 motif 1089:1093 the characteristic calcium binding motif 1054:1093 the characteristic calcium binding motif found in the other angiopoietins 1054:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 8 26 theme human 1191:1195 arg1 ANGPTL3 1197:1203 human ANGPTL3 1191:1203 human ANGPTL3 1191:1203 By radiation hybrid mapping and the use of surrounding genes, human ANGPTL3 maps to the 1p31 region. 10644446 5 27 theme Murine 693:698 arg1 polypeptide 722:732 a 455-acid polypeptide 711:732 a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA 711:806 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 5 27 theme Murine 693:698 arg1 ANGPTL3 700:706 Murine ANGPTL3 693:706 Murine ANGPTL3 693:706 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 4 28 theme 460-amino-acid 444:457 arg1 ANGPTL3 431:437 Human ANGPTL3 425:437 Human ANGPTL3 425:437 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 28 theme 460-amino-acid 444:457 arg1 polypeptide 459:469 a 460-amino-acid polypeptide 442:469 a 460-amino-acid polypeptide with the characteristic structure of angiopoietins 442:520 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 1 29 theme EST 113:115 arg1 databases 117:125 EST databases 113:125 EST databases for signal sequences and amphipathic helices 113:170 Based on searches of EST databases for signal sequences and amphipathic helices, we have identified and cloned an angiopoietin-like gene, ANGPTL3. 10644446 4 30 theme linker 628:633 arg1 peptide 635:641 a short linker peptide 620:641 a short linker peptide 620:641 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 3 31 theme adult 412:416 arg1 liver 418:422 adult liver 412:422 adult liver 412:422 ANGPTL3 is expressed early during liver development, and expression is maintained in adult liver. 10644446 2 32 theme Multiple 239:246 arg1 blots 264:268 Multiple tissue Northern blots 239:268 Multiple tissue Northern blots 239:268 Multiple tissue Northern blots show that ANGPTL3 is expressed principally in the liver. 10644446 8 33 theme surrounding 1172:1182 arg1 genes 1184:1188 surrounding genes 1172:1188 surrounding genes 1172:1188 By radiation hybrid mapping and the use of surrounding genes, human ANGPTL3 maps to the 1p31 region. 10644446 1 34 theme angiopoietin-like 206:222 arg1 ANGPTL3 230:236 ANGPTL3 230:236 ANGPTL3 230:236 Based on searches of EST databases for signal sequences and amphipathic helices, we have identified and cloned an angiopoietin-like gene, ANGPTL3. 10644446 1 34 theme angiopoietin-like 206:222 arg1 gene 224:227 an angiopoietin-like gene 203:227 an angiopoietin-like gene 203:227 Based on searches of EST databases for signal sequences and amphipathic helices, we have identified and cloned an angiopoietin-like gene, ANGPTL3. 10644446 0 35 theme angiopoietin-related 30:49 arg1 protein 51:57 a mammalian angiopoietin-related protein 18:57 a mammalian angiopoietin-related protein 18:57 Identification of a mammalian angiopoietin-related protein expressed specifically in liver. 10644446 8 36 theme hybrid 1142:1147 arg1 mapping 1149:1155 radiation hybrid mapping 1132:1155 radiation hybrid mapping 1132:1155 By radiation hybrid mapping and the use of surrounding genes, human ANGPTL3 maps to the 1p31 region. 10644446 8 37 theme radiation 1132:1140 arg1 mapping 1149:1155 radiation hybrid mapping 1132:1155 radiation hybrid mapping 1132:1155 By radiation hybrid mapping and the use of surrounding genes, human ANGPTL3 maps to the 1p31 region. 10644446 7 38 theme characteristic 1058:1071 arg1 motif 1089:1093 the characteristic calcium binding motif 1054:1093 the characteristic calcium binding motif found in the other angiopoietins 1054:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 6 39 theme conserved 835:843 arg1 cysteines 845:853 the four conserved cysteines 826:853 the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD 826:917 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 1 40 theme databases 117:125 arg1 searches 101:108 searches 101:108 searches of EST databases for signal sequences and amphipathic helices 101:170 Based on searches of EST databases for signal sequences and amphipathic helices, we have identified and cloned an angiopoietin-like gene, ANGPTL3. 10644446 7 41 theme calcium 1073:1079 arg1 motif 1089:1093 the characteristic calcium binding motif 1054:1093 the characteristic calcium binding motif found in the other angiopoietins 1054:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 4 42 theme extended 544:551 arg1 domain 561:566 an extended helical domain 541:566 an extended helical domain predicted to form dimeric or trimeric coiled-coils 541:617 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 43 theme Human 425:429 arg1 ANGPTL3 431:437 Human ANGPTL3 425:437 Human ANGPTL3 425:437 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 4 43 theme Human 425:429 arg1 polypeptide 459:469 a 460-amino-acid polypeptide 442:469 a 460-amino-acid polypeptide with the characteristic structure of angiopoietins 442:520 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 6 44 contain contain 936:942 arg1 it 924:925 it 924:925 it 924:925 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 6 44 contain contain 936:942 arg2 cysteines 954:962 two other cysteines 944:962 two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4 944:1021 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 1 45 theme signal 131:136 arg1 sequences 138:146 signal sequences 131:146 signal sequences 131:146 Based on searches of EST databases for signal sequences and amphipathic helices, we have identified and cloned an angiopoietin-like gene, ANGPTL3. 10644446 5 46 theme mouse 760:764 arg1 chromosome 766:775 mouse chromosome 4 760:777 mouse chromosome 4 760:777 Murine ANGPTL3 is a 455-acid polypeptide encoded by seven exons on mouse chromosome 4, spanning about 11 kb of DNA. 10644446 7 47 contain contain 1046:1052 arg1 ANGPTL3 1024:1030 ANGPTL3 1024:1030 ANGPTL3 1024:1030 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 7 47 contain contain 1046:1052 arg2 motif 1089:1093 the characteristic calcium binding motif 1054:1093 the characteristic calcium binding motif found in the other angiopoietins 1054:1126 ANGPTL3 also does not contain the characteristic calcium binding motif found in the other angiopoietins. 10644446 2 48 theme Northern 255:262 arg1 blots 264:268 Multiple tissue Northern blots 239:268 Multiple tissue Northern blots 239:268 Multiple tissue Northern blots show that ANGPTL3 is expressed principally in the liver. 10644446 4 49 theme short 622:626 arg1 peptide 635:641 a short linker peptide 620:641 a short linker peptide 620:641 Human ANGPTL3 is a 460-amino-acid polypeptide with the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen homology domain (FHD). 10644446 6 50 theme other 948:952 arg1 cysteines 954:962 two other cysteines 944:962 two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4 944:1021 ANGPTL3 contains the four conserved cysteines implicated in the intramolecular disulfide bonds within the FHD, but it does not contain two other cysteines that are found within the FHD of angiopoietins 1, 2, and 4. 10644446 2 51 theme tissue 248:253 arg1 blots 264:268 Multiple tissue Northern blots 239:268 Multiple tissue Northern blots 239:268 Multiple tissue Northern blots show that ANGPTL3 is expressed principally in the liver. 10644446 1 52 theme amphipathic 152:162 arg1 helices 164:170 amphipathic helices 152:170 amphipathic helices 152:170 Based on searches of EST databases for signal sequences and amphipathic helices, we have identified and cloned an angiopoietin-like gene, ANGPTL3. 1883960 2 0 theme 25-kD 553:557 arg1 species 600:606 25-kD and 28- to 30-kD fibroblast-derived IL-6 species 553:606 25-kD and 28- to 30-kD fibroblast-derived IL-6 species 553:606 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 4 1 theme O-glycosylated 1037:1050 arg1 IL-6 1052:1055 25-kD O-glycosylated IL-6 1031:1055 25-kD O-glycosylated IL-6 1031:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 5 2 theme 21-kD 1568:1572 arg1 species 1574:1580 a major apparently unglycosylated 21-kD species 1534:1580 a major apparently unglycosylated 21-kD species 1534:1580 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 3 theme peptide 1329:1335 arg1 site 1346:1349 the signal peptide cleavage site 1318:1349 the signal peptide cleavage site 1318:1349 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 5 4 theme Epstein-Barr 1404:1415 arg1 virus 1417:1421 the Epstein-Barr virus 1400:1421 the Epstein-Barr virus (EBV) 1400:1427 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 5 4 theme Epstein-Barr 1404:1415 arg1 EBV 1424:1426 EBV 1424:1426 EBV 1424:1426 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 2 5 theme 30-kD 570:574 arg1 IL-6 595:598 30-kD fibroblast-derived IL-6 570:598 30-kD fibroblast-derived IL-6 570:598 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 2 6 theme immunoaffinity 697:710 arg1 chromatography 712:725 immunoaffinity chromatography 697:725 immunoaffinity chromatography 697:725 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 1 7 theme 23- 448:450 arg1 set 475:477 a set 473:477 a set of at least three N- and O-glycosylated 28- to 30-kD species 473:538 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 7 theme 23- 448:450 arg1 set 411:413 a set 409:413 a set of at least three O-glycosylated 23- to 25-kD species 409:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 7 theme 23- 448:450 arg1 23- 448:450 at least three O-glycosylated 23- 418:450 at least three O-glycosylated 23- to 25-kD species 418:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 8 theme fibroblast-derived 1005:1022 arg1 23- 1024:1026 the fibroblast-derived 23- 1001:1026 the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 1001:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 9 theme endometrial 188:198 arg1 cells 208:212 endometrial stromal cells 188:212 endometrial stromal cells 188:212 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 2 10 theme 28- 563:565 arg1 species 600:606 25-kD and 28- to 30-kD fibroblast-derived IL-6 species 553:606 25-kD and 28- to 30-kD fibroblast-derived IL-6 species 553:606 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 4 11 theme 30-kD 1117:1121 arg1 N- 1123:1124 30-kD N- 1117:1124 30-kD N- 1117:1124 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 5 12 theme N-glycosylated 1600:1613 arg1 species 1615:1621 a minor 25-kD N-glycosylated species 1586:1621 a minor 25-kD N-glycosylated species 1586:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 1 13 theme 25-kD 455:459 arg1 species 461:467 25-kD species 455:467 25-kD species 455:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 14 dep primate 1448:1454 arg1 tamarin 1457:1463 tamarin 1457:1463 tamarin 1457:1463 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 15 theme amino 1088:1092 arg1 terminus 1094:1101 the major amino terminus 1078:1101 the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 1078:1148 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 16 theme digestion 740:748 arg1 experiments 750:760 Glycosidase digestion experiments 728:760 Glycosidase digestion experiments on such purified preparations 728:790 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 5 17 theme minor 1588:1592 arg1 species 1615:1621 a minor 25-kD N-glycosylated species 1586:1621 a minor 25-kD N-glycosylated species 1586:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 18 gly O-glycosylated 1037:1050 arg1 IL-6 1052:1055 25-kD O-glycosylated IL-6 1031:1055 25-kD O-glycosylated IL-6 1031:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 19 dep conditions 282:291 arg1 consists 294:301 consists 294:301 consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 294:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 3 20 theme IL-6 843:846 arg1 species 848:854 almost all human fibroblast-derived IL-6 species 807:854 almost all human fibroblast-derived IL-6 species 807:854 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 3 21 link fibroblast-derived 824:841 arg1 species 848:854 almost all human fibroblast-derived IL-6 species 807:854 almost all human fibroblast-derived IL-6 species 807:854 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 3 22 theme purified 770:777 arg1 preparations 779:790 such purified preparations 765:790 such purified preparations 765:790 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 3 23 theme human 818:822 arg1 species 848:854 almost all human fibroblast-derived IL-6 species 807:854 almost all human fibroblast-derived IL-6 species 807:854 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 2 24 theme combination 671:681 arg1 use 662:664 the use 658:664 the use of a combination of lectin and immunoaffinity chromatography 658:725 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 1 25 theme phosphoglycoproteins 339:358 arg1 phosphoglycoproteins 339:358 differentially modified phosphoglycoproteins 315:358 differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 315:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 25 theme phosphoglycoproteins 339:358 arg1 set 308:310 a set 306:310 a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 306:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 26 theme major 977:981 arg1 terminus 989:996 the major amino terminus 973:996 the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 973:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 0 27 from differences 26:36 arg1 glycosylation 41:53 glycosylation 41:53 glycosylation of human interleukin-6 41:76 Marked cell-type-specific differences in glycosylation of human interleukin-6. 1883960 5 28 theme primate 1448:1454 arg1 lines 1473:1477 -infected human and primate (tamarin) B-cell lines 1428:1477 -infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species 1428:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 1 29 theme molecular 363:371 arg1 mass 373:376 molecular mass 363:376 molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 363:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 30 dep " 1386:1386 arg1 secreted 1388:1395 secreted 1388:1395 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species 1371:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 31 theme Amino 937:941 arg1 sequencing 948:957 Amino acid sequencing 937:957 Amino acid sequencing 937:957 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 4 32 theme synthesized 1199:1209 arg1 polypeptides 1216:1227 newly synthesized IL-6 polypeptides 1193:1227 newly synthesized IL-6 polypeptides 1193:1227 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 33 from kD 405:406 arg1 mass 373:376 molecular mass 363:376 molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 363:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 33 from kD 405:406 arg1 range 385:389 the range 381:389 the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 381:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 34 theme Human 79:83 arg1 interleukin-6 85:97 Human interleukin-6 79:97 Human interleukin-6 (IL-6) 79:104 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 34 theme Human 79:83 arg1 IL-6 100:103 IL-6 100:103 IL-6 100:103 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 35 from differences 1303:1313 arg1 site 1346:1349 the signal peptide cleavage site 1318:1349 the signal peptide cleavage site 1318:1349 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 2 36 theme fibroblast-derived 576:593 arg1 IL-6 595:598 30-kD fibroblast-derived IL-6 570:598 30-kD fibroblast-derived IL-6 570:598 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 5 37 theme human 1438:1442 arg1 lines 1473:1477 -infected human and primate (tamarin) B-cell lines 1428:1477 -infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species 1428:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 0 38 theme Marked 0:5 arg1 differences 26:36 Marked cell-type-specific differences 0:36 Marked cell-type-specific differences in glycosylation of human interleukin-6. 0:77 Marked cell-type-specific differences in glycosylation of human interleukin-6. 1883960 4 39 theme 25-kD 1031:1035 arg1 IL-6 1052:1055 25-kD O-glycosylated IL-6 1031:1055 25-kD O-glycosylated IL-6 1031:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 40 theme 30-kD 902:906 arg1 species 908:914 30-kD species 902:914 30-kD species 902:914 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 4 41 theme 28- 1110:1112 arg1 IL-6 1145:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 42 from mass 373:376 arg1 kD 405:406 23 to 30 kD 396:406 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 396:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 42 from mass 373:376 arg1 range 385:389 the range 381:389 the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 381:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 43 theme B-cell 1466:1471 arg1 lines 1473:1477 -infected human and primate (tamarin) B-cell lines 1428:1477 -infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species 1428:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 0 44 theme human 58:62 arg1 interleukin-6 64:76 human interleukin-6 58:76 human interleukin-6 58:76 Marked cell-type-specific differences in glycosylation of human interleukin-6. 1883960 4 45 from terminus 989:996 arg1 23- 1024:1026 the fibroblast-derived 23- 1001:1026 the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 1001:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 5 46 gly unglycosylated 1553:1566 arg1 species 1574:1580 a major apparently unglycosylated 21-kD species 1534:1580 a major apparently unglycosylated 21-kD species 1534:1580 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 47 theme major 1082:1086 arg1 terminus 1094:1101 the major amino terminus 1078:1101 the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 1078:1148 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 48 dep kD 405:406 arg1 set 475:477 a set 473:477 a set of at least three N- and O-glycosylated 28- to 30-kD species 473:538 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 48 dep kD 405:406 arg1 set 411:413 a set 409:413 a set of at least three O-glycosylated 23- to 25-kD species 409:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 48 dep kD 405:406 arg1 23- 448:450 at least three O-glycosylated 23- 418:450 at least three O-glycosylated 23- to 25-kD species 418:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 49 theme major 1536:1540 arg1 species 1574:1580 a major apparently unglycosylated 21-kD species 1534:1580 a major apparently unglycosylated 21-kD species 1534:1580 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 50 theme different 1242:1250 arg1 pathways 1263:1270 the two different processing pathways 1234:1270 the two different processing pathways in fibroblasts 1234:1285 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 51 from phosphoglycoproteins 339:358 arg1 range 385:389 the range 381:389 the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 381:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 52 theme unglycosylated 1553:1566 arg1 species 1574:1580 a major apparently unglycosylated 21-kD species 1534:1580 a major apparently unglycosylated 21-kD species 1534:1580 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 1 53 theme denaturing 258:267 arg1 conditions 282:291 reducing conditions 273:291 reducing conditions 273:291 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 54 theme signal 1322:1327 arg1 site 1346:1349 the signal peptide cleavage site 1318:1349 the signal peptide cleavage site 1318:1349 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 55 theme reducing 273:280 arg1 conditions 282:291 reducing conditions 273:291 reducing conditions 273:291 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 2 56 theme chromatography 712:725 arg1 combination 671:681 a combination 669:681 a combination of lectin and immunoaffinity chromatography 669:725 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 2 57 theme lectin 686:691 arg1 combination 671:681 a combination 669:681 a combination of lectin and immunoaffinity chromatography 669:725 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 1 58 theme O-glycosylated 433:446 arg1 23- 448:450 at least three O-glycosylated 23- 418:450 at least three O-glycosylated 23- to 25-kD species 418:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 59 gly phosphoglycoproteins 339:358 arg1 phosphoglycoproteins 339:358 differentially modified phosphoglycoproteins 315:358 differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 315:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 60 theme 25-kD 1594:1598 arg1 species 1615:1621 a minor 25-kD N-glycosylated species 1586:1621 a minor 25-kD N-glycosylated species 1586:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 1 61 theme stromal 200:206 arg1 cells 208:212 endometrial stromal cells 188:212 endometrial stromal cells 188:212 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 5 62 gly N-glycosylated 1600:1613 arg1 species 1615:1621 a minor 25-kD N-glycosylated species 1586:1621 a minor 25-kD N-glycosylated species 1586:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 4 63 from terminus 1094:1101 arg1 IL-6 1145:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 64 gly N-glycosylated 921:934 arg1 28- 895:897 28- 895:897 28- 895:897 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 3 64 gly N-glycosylated 921:934 arg1 N-glycosylated 921:934 N-glycosylated 921:934 N-glycosylated 921:934 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 4 65 theme cleavage 1337:1344 arg1 site 1346:1349 the signal peptide cleavage site 1318:1349 the signal peptide cleavage site 1318:1349 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 66 theme Glycosidase 728:738 arg1 experiments 750:760 Glycosidase digestion experiments 728:760 Glycosidase digestion experiments on such purified preparations 728:790 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 1 67 theme endotoxin-induced 131:147 arg1 fibroblasts 149:159 endotoxin-induced fibroblasts 131:159 endotoxin-induced fibroblasts 131:159 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 68 theme O-glycosylated 1130:1143 arg1 IL-6 1145:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 69 theme fibroblast-derived 824:841 arg1 species 848:854 almost all human fibroblast-derived IL-6 species 807:854 almost all human fibroblast-derived IL-6 species 807:854 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 1 70 gly O-glycosylated 433:446 arg1 23- 448:450 at least three O-glycosylated 23- 418:450 at least three O-glycosylated 23- to 25-kD species 418:467 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 3 71 theme such 765:768 arg1 preparations 779:790 such purified preparations 765:790 such purified preparations 765:790 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 4 72 theme IL-6 1211:1214 arg1 polypeptides 1216:1227 newly synthesized IL-6 polypeptides 1193:1227 newly synthesized IL-6 polypeptides 1193:1227 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 73 from experiments 750:760 arg1 preparations 779:790 such purified preparations 765:790 such purified preparations 765:790 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 3 74 theme all 814:816 arg1 species 848:854 almost all human fibroblast-derived IL-6 species 807:854 almost all human fibroblast-derived IL-6 species 807:854 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 1 75 theme mass 373:376 arg1 phosphoglycoproteins 339:358 differentially modified phosphoglycoproteins 315:358 differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 315:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 76 link fibroblast-derived 1005:1022 arg1 23- 1024:1026 the fibroblast-derived 23- 1001:1026 the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 1001:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 3 77 gly O-glycosylated 861:874 arg1 species 848:854 almost all human fibroblast-derived IL-6 species 807:854 almost all human fibroblast-derived IL-6 species 807:854 Glycosidase digestion experiments on such purified preparations confirmed that almost all human fibroblast-derived IL-6 species were O-glycosylated; additionally, the 28- to 30-kD species were N-glycosylated. 1883960 4 78 theme acid 943:946 arg1 sequencing 948:957 Amino acid sequencing 937:957 Amino acid sequencing 937:957 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 5 79 theme -infected 1428:1436 arg1 lines 1473:1477 -infected human and primate (tamarin) B-cell lines 1428:1477 -infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species 1428:1621 Unexpectedly, IL-6 "constitutively" secreted by the Epstein-Barr virus (EBV)-infected human and primate (tamarin) B-cell lines designated sfBJAB and sfBT, respectively, consisted of a major apparently unglycosylated 21-kD species and a minor 25-kD N-glycosylated species. 1883960 0 80 theme cell-type-specific 7:24 arg1 differences 26:36 Marked cell-type-specific differences 0:36 Marked cell-type-specific differences in glycosylation of human interleukin-6. 0:77 Marked cell-type-specific differences in glycosylation of human interleukin-6. 1883960 2 81 link fibroblast-derived 576:593 arg1 IL-6 595:598 30-kD fibroblast-derived IL-6 570:598 30-kD fibroblast-derived IL-6 570:598 The 23- to 25-kD and 28- to 30-kD fibroblast-derived IL-6 species have been separately purified to homogeneity with the use of a combination of lectin and immunoaffinity chromatography. 1883960 1 82 theme 30-kD 526:530 arg1 species 532:538 30-kD species 526:538 30-kD species 526:538 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 83 theme polypeptides 1216:1227 arg1 targeting 1180:1188 targeting 1180:1188 targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts 1180:1285 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 84 dep N- 497:498 arg1 28- 519:521 28- 519:521 28- 519:521 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 84 dep N- 497:498 arg1 three 491:495 three 491:495 three 491:495 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 1 85 theme endothelial 219:229 arg1 cells 231:235 endothelial cells 219:235 endothelial cells 219:235 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 86 theme amino 983:987 arg1 terminus 989:996 the major amino terminus 973:996 the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 973:1055 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 4 87 from pathways 1263:1270 arg1 fibroblasts 1275:1285 fibroblasts 1275:1285 fibroblasts 1275:1285 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 0 88 theme interleukin-6 64:76 arg1 glycosylation 41:53 glycosylation 41:53 glycosylation of human interleukin-6 41:76 Marked cell-type-specific differences in glycosylation of human interleukin-6. 1883960 4 89 gly O-glycosylated 1130:1143 arg1 IL-6 1145:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 the 28- to 30-kD N- and O-glycosylated IL-6 1106:1148 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 90 theme modified 330:337 arg1 phosphoglycoproteins 339:358 differentially modified phosphoglycoproteins 315:358 differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 315:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 0 91 gly glycosylation 41:53 arg1 interleukin-6 64:76 human interleukin-6 58:76 human interleukin-6 58:76 Marked cell-type-specific differences in glycosylation of human interleukin-6. 1883960 1 92 dep 30 402:403 arg1 to 399:400 to 399:400 to 399:400 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 1883960 4 93 theme processing 1252:1261 arg1 pathways 1263:1270 the two different processing pathways 1234:1270 the two different processing pathways in fibroblasts 1234:1285 Amino acid sequencing revealed that the major amino terminus in the fibroblast-derived 23- to 25-kD O-glycosylated IL-6 was at Ala28 whereas the major amino terminus in the 28- to 30-kD N- and O-glycosylated IL-6 was at Val30, suggesting that targeting of newly synthesized IL-6 polypeptides into the two different processing pathways in fibroblasts may be keyed to differences in the signal peptide cleavage site. 1883960 1 94 from range 385:389 arg1 phosphoglycoproteins 339:358 differentially modified phosphoglycoproteins 315:358 differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species) 315:539 Human interleukin-6 (IL-6) secreted by cytokine- or endotoxin-induced fibroblasts, monocytes, keratinocytes, endometrial stromal cells, and endothelial cells, when analyzed under denaturing and reducing conditions, consists of a set of differentially modified phosphoglycoproteins of molecular mass in the range from 23 to 30 kD (a set of at least three O-glycosylated 23- to 25-kD species and a set of at least three N- and O-glycosylated 28- to 30-kD species). 11152692 10 0 theme mass 1141:1144 arg1 spectrometry 1146:1157 mass spectrometry 1141:1157 mass spectrometry of the isolated protein resulting in 84% sequence coverage 1141:1216 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 6 1 contain contains 597:604 arg1 asporin 589:595 asporin 589:595 asporin 589:595 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 6 1 contain contains 597:604 arg2 stretch 615:621 a unique stretch 606:621 a unique stretch of aspartic acid residues in its amino-terminal region 606:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 9 2 theme tissues 982:988 arg1 variety 965:971 a variety 963:971 a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver 963:1078 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 9 2 theme tissues 982:988 arg1 tissues 982:988 human tissues 976:988 human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver 976:1078 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 3 3 theme mouse 306:310 arg1 cDNAs 320:324 human and mouse asporin cDNAs 296:324 human and mouse asporin cDNAs 296:324 Cloning of human and mouse asporin cDNAs revealed that the protein is closely related to decorin and biglycan. 11152692 2 4 theme leucine-rich 177:188 arg1 family 197:202 the leucine-rich repeat family 173:202 the leucine-rich repeat family of proteins 173:214 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 12 5 theme aspartate-rich 1402:1415 arg1 terminus 1423:1430 the aspartate-rich amino terminus 1398:1430 the aspartate-rich amino terminus 1398:1430 The name asporin reflects the aspartate-rich amino terminus and the overall similarity to decorin. 11152692 8 6 theme human 803:807 arg1 gene 817:820 the human asporin gene 799:820 the human asporin gene 799:820 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 9 7 with tissues 982:988 arg1 levels 1002:1007 higher levels 995:1007 higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver 995:1078 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 8 8 theme consensus 909:917 arg1 sequences 919:927 TATA consensus sequences 904:927 TATA consensus sequences 904:927 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 9 9 theme higher 995:1000 arg1 levels 1002:1007 higher levels 995:1007 higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver 995:1078 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 10 10 theme asporin 1116:1122 arg1 sequence 1104:1111 The deduced amino acid sequence 1081:1111 The deduced amino acid sequence of asporin 1081:1122 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 8 11 theme gene 817:820 arg1 exons 790:794 The 8 exons 784:794 The 8 exons of the human asporin gene 784:820 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 11 12 theme O-glycosylation 1339:1353 arg1 site 1355:1358 a potential O-glycosylation site 1327:1358 a potential O-glycosylation site at Ser(54) 1327:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 13 theme oligosaccharide 1297:1311 arg1 structure 1313:1321 a heterogeneous oligosaccharide structure 1281:1321 a heterogeneous oligosaccharide structure 1281:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 10 14 theme sequence 1200:1207 arg1 coverage 1209:1216 84% sequence coverage 1196:1216 84% sequence coverage 1196:1216 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 10 15 theme protein 1175:1181 arg1 spectrometry 1146:1157 mass spectrometry 1141:1157 mass spectrometry of the isolated protein resulting in 84% sequence coverage 1141:1216 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 11 16 from Asn 1267:1269 arg1 site 1355:1358 a potential O-glycosylation site 1327:1358 a potential O-glycosylation site at Ser(54) 1327:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 16 from Asn 1267:1269 arg1 site 1259:1262 an N-glycosylation site 1240:1262 an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure 1240:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 8 17 theme asporin 809:815 arg1 gene 817:820 the human asporin gene 799:820 the human asporin gene 799:820 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 10 18 theme acid 1099:1102 arg1 sequence 1104:1111 The deduced amino acid sequence 1081:1111 The deduced amino acid sequence of asporin 1081:1122 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 4 19 theme leucine-rich 462:473 arg1 propeptide 419:428 a putative propeptide 408:428 a putative propeptide 408:428 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 19 theme leucine-rich 462:473 arg1 repeats 475:481 10 leucine-rich repeats 459:481 10 leucine-rich repeats 459:481 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 9 20 theme asporin 934:940 arg1 mRNA 942:945 The asporin mRNA 930:945 The asporin mRNA 930:945 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 1 21 theme leucine-rich 70:81 arg1 family 98:103 the leucine-rich repeat protein family 66:103 the leucine-rich repeat protein family closely related to decorin and biglycan 66:143 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. 11152692 6 22 theme amino-terminal 656:669 arg1 region 671:676 its amino-terminal region 652:676 its amino-terminal region 652:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 11 23 theme N-glycosylation 1243:1257 arg1 site 1259:1262 an N-glycosylation site 1240:1262 an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure 1240:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 24 gly O-glycosylation 1339:1353 arg2 281 1271:1273 281 1271:1273 281 1271:1273 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 24 gly O-glycosylation 1339:1353 arg2 Asn 1267:1269 Asn 1267:1269 Asn(281) 1267:1274 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 24 gly O-glycosylation 1339:1353 arg2 54 1367:1368 54 1367:1368 54 1367:1368 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 24 gly O-glycosylation 1339:1353 arg2 Ser 1363:1365 Ser 1363:1365 Ser(54) 1363:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 24 gly O-glycosylation 1339:1353 arg2 site 1355:1358 a potential O-glycosylation site 1327:1358 a potential O-glycosylation site at Ser(54) 1327:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 1 25 theme repeat 83:88 arg1 family 98:103 the leucine-rich repeat protein family 66:103 the leucine-rich repeat protein family closely related to decorin and biglycan 66:143 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. 11152692 9 26 theme articular 1027:1035 arg1 cartilage 1037:1045 osteoarthritic articular cartilage 1012:1045 osteoarthritic articular cartilage 1012:1045 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 12 27 theme overall 1440:1446 arg1 similarity 1448:1457 the overall similarity 1436:1457 the overall similarity to decorin 1436:1468 The name asporin reflects the aspartate-rich amino terminus and the overall similarity to decorin. 11152692 11 28 theme heterogeneous 1283:1295 arg1 structure 1313:1321 a heterogeneous oligosaccharide structure 1281:1321 a heterogeneous oligosaccharide structure 1281:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 1 29 theme protein 90:96 arg1 family 98:103 the leucine-rich repeat protein family 66:103 the leucine-rich repeat protein family closely related to decorin and biglycan 66:143 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. 11152692 4 30 contain contains 399:406 arg2 cysteines 448:456 4 amino-terminal cysteines 431:456 4 amino-terminal cysteines 431:456 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 30 contain contains 399:406 arg1 It 396:397 It 396:397 It 396:397 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 30 contain contains 399:406 arg2 propeptide 419:428 a putative propeptide 408:428 a putative propeptide 408:428 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 30 contain contains 399:406 arg2 cysteines 501:509 2 C-terminal cysteines 488:509 2 C-terminal cysteines 488:509 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 30 contain contains 399:406 arg2 repeats 475:481 10 leucine-rich repeats 459:481 10 leucine-rich repeats 459:481 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 1 31 theme family 98:103 arg1 member 56:61 a novel member 48:61 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan 48:143 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. 11152692 6 32 theme residues 640:647 arg1 stretch 615:621 a unique stretch 606:621 a unique stretch of aspartic acid residues in its amino-terminal region 606:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 11 33 gly N-glycosylation 1243:1257 arg2 281 1271:1273 281 1271:1273 281 1271:1273 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 33 gly N-glycosylation 1243:1257 arg2 Asn 1267:1269 Asn 1267:1269 Asn(281) 1267:1274 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 33 gly N-glycosylation 1243:1257 arg2 54 1367:1368 54 1367:1368 54 1367:1368 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 33 gly N-glycosylation 1243:1257 arg2 site 1259:1262 an N-glycosylation site 1240:1262 an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure 1240:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 33 gly N-glycosylation 1243:1257 arg2 Ser 1363:1365 Ser 1363:1365 Ser(54) 1363:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 6 34 from residues 640:647 arg1 region 671:676 its amino-terminal region 652:676 its amino-terminal region 652:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 6 35 theme acid 635:638 arg1 residues 640:647 aspartic acid residues 626:647 aspartic acid residues in its amino-terminal region 626:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 8 36 from kilobases 830:838 arg1 chromosome 843:852 chromosome 9q31.1-32 843:862 chromosome 9q31.1-32 843:862 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 10 37 theme % 1198:1198 arg1 coverage 1209:1216 84% sequence coverage 1196:1216 84% sequence coverage 1196:1216 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 11 38 with site 1259:1262 arg1 structure 1313:1321 a heterogeneous oligosaccharide structure 1281:1321 a heterogeneous oligosaccharide structure 1281:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 10 39 theme isolated 1166:1173 arg1 protein 1175:1181 the isolated protein 1162:1181 the isolated protein resulting in 84% sequence coverage 1162:1216 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 11 40 from Ser 1363:1365 arg1 site 1355:1358 a potential O-glycosylation site 1327:1358 a potential O-glycosylation site at Ser(54) 1327:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 40 from Ser 1363:1365 arg1 site 1259:1262 an N-glycosylation site 1240:1262 an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure 1240:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 6 41 theme aspartic 626:633 arg1 residues 640:647 aspartic acid residues 626:647 aspartic acid residues in its amino-terminal region 626:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 0 42 theme asporin 39:45 arg1 characterization 19:34 characterization 19:34 characterization 19:34 Identification and characterization of asporin. 11152692 0 42 theme asporin 39:45 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and characterization of asporin. 11152692 7 43 theme aspartate 743:751 arg1 residues 753:760 consecutive aspartate residues 731:760 consecutive aspartate residues 731:760 A polymorphism was identified in that the number of consecutive aspartate residues varied from 11 to 15. 11152692 4 44 theme amino-terminal 433:446 arg1 cysteines 448:456 4 amino-terminal cysteines 431:456 4 amino-terminal cysteines 431:456 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 44 theme amino-terminal 433:446 arg1 propeptide 419:428 a putative propeptide 408:428 a putative propeptide 408:428 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 6 45 from region 671:676 arg1 stretch 615:621 a unique stretch 606:621 a unique stretch of aspartic acid residues in its amino-terminal region 606:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 10 46 theme 84 1196:1197 arg1 % 1198:1198 % 1198:1198 % 1198:1198 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 4 47 theme C-terminal 490:499 arg1 cysteines 501:509 2 C-terminal cysteines 488:509 2 C-terminal cysteines 488:509 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 47 theme C-terminal 490:499 arg1 propeptide 419:428 a putative propeptide 408:428 a putative propeptide 408:428 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 12 48 theme amino 1417:1421 arg1 terminus 1423:1430 the aspartate-rich amino terminus 1398:1430 the aspartate-rich amino terminus 1398:1430 The name asporin reflects the aspartate-rich amino terminus and the overall similarity to decorin. 11152692 10 49 theme deduced 1085:1091 arg1 sequence 1104:1111 The deduced amino acid sequence 1081:1111 The deduced amino acid sequence of asporin 1081:1122 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 2 50 theme proteins 207:214 arg1 family 197:202 the leucine-rich repeat family 173:202 the leucine-rich repeat family of proteins 173:214 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 1 51 theme related 113:119 arg1 family 98:103 the leucine-rich repeat protein family 66:103 the leucine-rich repeat protein family closely related to decorin and biglycan 66:143 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. 11152692 11 52 theme potential 1329:1337 arg1 site 1355:1358 a potential O-glycosylation site 1327:1358 a potential O-glycosylation site at Ser(54) 1327:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 3 53 theme human 296:300 arg1 cDNAs 320:324 human and mouse asporin cDNAs 296:324 human and mouse asporin cDNAs 296:324 Cloning of human and mouse asporin cDNAs revealed that the protein is closely related to decorin and biglycan. 11152692 2 54 theme family 197:202 arg1 member 163:168 a novel member 155:168 a novel member of the leucine-rich repeat family of proteins 155:214 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 2 54 theme family 197:202 arg1 Asporin 146:152 Asporin 146:152 Asporin 146:152 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 6 55 from stretch 615:621 arg1 region 671:676 its amino-terminal region 652:676 its amino-terminal region 652:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 8 56 theme putative 873:880 arg1 region 891:896 the putative promoter region 869:896 the putative promoter region 869:896 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 2 57 theme novel 157:161 arg1 member 163:168 a novel member 155:168 a novel member of the leucine-rich repeat family of proteins 155:214 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 2 57 theme novel 157:161 arg1 Asporin 146:152 Asporin 146:152 Asporin 146:152 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 7 58 theme consecutive 731:741 arg1 residues 753:760 consecutive aspartate residues 731:760 consecutive aspartate residues 731:760 A polymorphism was identified in that the number of consecutive aspartate residues varied from 11 to 15. 11152692 2 59 theme repeat 190:195 arg1 family 197:202 the leucine-rich repeat family 173:202 the leucine-rich repeat family of proteins 173:214 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 2 60 theme human 245:249 arg1 cartilage 261:269 human articular cartilage 245:269 human articular cartilage 245:269 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 11 61 with site 1355:1358 arg1 structure 1313:1321 a heterogeneous oligosaccharide structure 1281:1321 a heterogeneous oligosaccharide structure 1281:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 2 62 theme articular 251:259 arg1 cartilage 261:269 human articular cartilage 245:269 human articular cartilage 245:269 Asporin, a novel member of the leucine-rich repeat family of proteins, was partially purified from human articular cartilage and meniscus. 11152692 4 63 theme putative 410:417 arg1 cysteines 448:456 4 amino-terminal cysteines 431:456 4 amino-terminal cysteines 431:456 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 63 theme putative 410:417 arg1 repeats 475:481 10 leucine-rich repeats 459:481 10 leucine-rich repeats 459:481 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 63 theme putative 410:417 arg1 cysteines 501:509 2 C-terminal cysteines 488:509 2 C-terminal cysteines 488:509 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 4 63 theme putative 410:417 arg1 propeptide 419:428 a putative propeptide 408:428 a putative propeptide 408:428 It contains a putative propeptide, 4 amino-terminal cysteines, 10 leucine-rich repeats, and 2 C-terminal cysteines. 11152692 11 64 contain contains 1231:1238 arg2 site 1355:1358 a potential O-glycosylation site 1327:1358 a potential O-glycosylation site at Ser(54) 1327:1369 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 64 contain contains 1231:1238 arg2 site 1259:1262 an N-glycosylation site 1240:1262 an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure 1240:1321 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 11 64 contain contains 1231:1238 arg1 protein 1223:1229 The protein 1219:1229 The protein 1219:1229 The protein contains an N-glycosylation site at Asn(281) with a heterogeneous oligosaccharide structure and a potential O-glycosylation site at Ser(54). 11152692 7 65 dep 15 780:781 arg1 to 777:778 to 777:778 to 777:778 A polymorphism was identified in that the number of consecutive aspartate residues varied from 11 to 15. 11152692 6 66 theme unique 608:613 arg1 stretch 615:621 a unique stretch 606:621 a unique stretch of aspartic acid residues in its amino-terminal region 606:676 Instead, asporin contains a unique stretch of aspartic acid residues in its amino-terminal region. 11152692 9 67 theme osteoarthritic 1012:1025 arg1 cartilage 1037:1045 osteoarthritic articular cartilage 1012:1045 osteoarthritic articular cartilage 1012:1045 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 3 68 theme asporin 312:318 arg1 cDNAs 320:324 human and mouse asporin cDNAs 296:324 human and mouse asporin cDNAs 296:324 Cloning of human and mouse asporin cDNAs revealed that the protein is closely related to decorin and biglycan. 11152692 12 69 theme name 1376:1379 arg1 asporin 1381:1387 The name asporin 1372:1387 The name asporin 1372:1387 The name asporin reflects the aspartate-rich amino terminus and the overall similarity to decorin. 11152692 9 70 from levels 1002:1007 arg1 heart 1063:1067 heart 1063:1067 heart 1063:1067 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 9 70 from levels 1002:1007 arg1 liver 1074:1078 liver 1074:1078 liver 1074:1078 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 9 70 from levels 1002:1007 arg1 uterus 1055:1060 uterus 1055:1060 uterus 1055:1060 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 9 70 from levels 1002:1007 arg1 aorta 1048:1052 aorta 1048:1052 aorta 1048:1052 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 9 70 from levels 1002:1007 arg1 cartilage 1037:1045 osteoarthritic articular cartilage 1012:1045 osteoarthritic articular cartilage 1012:1045 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 8 71 theme promoter 882:889 arg1 region 891:896 the putative promoter region 869:896 the putative promoter region 869:896 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 5 72 from proteoglycan 566:577 arg1 contrast 515:522 contrast 515:522 contrast to decorin and biglycan 515:546 In contrast to decorin and biglycan, asporin is not a proteoglycan. 11152692 10 73 theme amino 1093:1097 arg1 sequence 1104:1111 The deduced amino acid sequence 1081:1111 The deduced amino acid sequence of asporin 1081:1122 The deduced amino acid sequence of asporin was confirmed by mass spectrometry of the isolated protein resulting in 84% sequence coverage. 11152692 3 74 theme cDNAs 320:324 arg1 Cloning 285:291 Cloning 285:291 Cloning of human and mouse asporin cDNAs 285:324 Cloning of human and mouse asporin cDNAs revealed that the protein is closely related to decorin and biglycan. 11152692 1 75 theme novel 50:54 arg1 member 56:61 a novel member 48:61 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan 48:143 a novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. 11152692 8 76 theme TATA 904:907 arg1 sequences 919:927 TATA consensus sequences 904:927 TATA consensus sequences 904:927 The 8 exons of the human asporin gene span 26 kilobases on chromosome 9q31.1-32, and the putative promoter region lacks TATA consensus sequences. 11152692 9 77 theme human 976:980 arg1 tissues 982:988 human tissues 976:988 human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver 976:1078 The asporin mRNA is expressed in a variety of human tissues with higher levels in osteoarthritic articular cartilage, aorta, uterus, heart, and liver. 11152692 7 78 theme residues 753:760 arg1 number 721:726 the number 717:726 the number of consecutive aspartate residues 717:760 A polymorphism was identified in that the number of consecutive aspartate residues varied from 11 to 15. 30089812 5 0 theme human 652:656 arg1 LH3 658:660 full-length human LH3 640:660 full-length human LH3 640:660 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 9 1 theme collagen-related 1199:1214 arg1 diseases 1216:1223 collagen-related diseases 1199:1223 collagen-related diseases involving human lysyl hydroxylases 1199:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 5 2 theme donor 692:696 arg1 substrates 698:707 donor substrates 692:707 donor substrates 692:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 8 3 theme catalytic 1047:1055 arg1 sites 1057:1061 the catalytic sites 1043:1061 the catalytic sites 1043:1061 Known disease-related mutations map in close proximity to the catalytic sites. 30089812 1 4 theme essential 177:185 arg1 roles 187:191 essential roles 177:191 essential roles in extracellular matrix (ECM) maturation and remodeling 177:247 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 0 5 theme LH3 97:99 arg1 architecture 10:21 Molecular architecture 0:21 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 0:100 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 4 6 theme molecular 552:560 arg1 information 572:582 molecular structure information 552:582 molecular structure information 552:582 Our understanding of LH3 functions is currently hampered by lack of molecular structure information. 30089812 5 7 theme full-length 640:650 arg1 LH3 658:660 full-length human LH3 640:660 full-length human LH3 640:660 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 9 8 theme LH3 1162:1164 arg1 mechanisms 1185:1194 the molecular mechanisms 1171:1194 the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases 1171:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 9 8 theme LH3 1162:1164 arg1 functions 1149:1157 the multiple functions 1136:1157 the multiple functions of LH3 1136:1164 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 3 9 theme collagen 413:420 arg1 hydroxylation 429:441 collagen lysine hydroxylation 413:441 collagen lysine hydroxylation 413:441 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 1 10 from roles 187:191 arg1 remodeling 238:247 remodeling 238:247 remodeling 238:247 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 1 10 from roles 187:191 arg1 maturation 223:232 extracellular matrix (ECM) maturation 196:232 extracellular matrix (ECM) maturation 196:232 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 9 11 theme structural 1100:1109 arg1 framework 1111:1119 a structural framework 1098:1119 a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases 1098:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 7 12 theme other 951:955 arg1 glycosyltransferases 963:982 other known glycosyltransferases 951:982 other known glycosyltransferases 951:982 The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. 30089812 8 13 theme close 1024:1028 arg1 proximity 1030:1038 close proximity 1024:1038 close proximity to the catalytic sites 1024:1061 Known disease-related mutations map in close proximity to the catalytic sites. 30089812 5 14 theme LH3 658:660 arg1 structures 626:635 high resolution crystal structures 602:635 high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates 602:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 6 15 from N- 795:796 arg1 sites 782:786 two distinct catalytic sites 759:786 two distinct catalytic sites 759:786 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 6 16 theme catalytic 772:780 arg1 sites 782:786 two distinct catalytic sites 759:786 two distinct catalytic sites 759:786 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 7 17 theme distinguishing 915:928 arg1 features 930:937 distinguishing features 915:937 distinguishing features 915:937 The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. 30089812 3 18 theme multiple 366:373 arg1 activities 385:394 multiple enzymatic activities 366:394 multiple enzymatic activities 366:394 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 4 19 theme functions 509:517 arg1 understanding 488:500 Our understanding 484:500 Our understanding of LH3 functions 484:517 Our understanding of LH3 functions is currently hampered by lack of molecular structure information. 30089812 2 20 theme tissue 304:309 arg1 disorders 311:319 severe connective tissue disorders 286:319 severe connective tissue disorders 286:319 Malfunctions in these enzymes cause severe connective tissue disorders. 30089812 6 21 theme distinct 763:770 arg1 sites 782:786 two distinct catalytic sites 759:786 two distinct catalytic sites 759:786 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 3 22 theme enzymatic 375:383 arg1 activities 385:394 multiple enzymatic activities 366:394 multiple enzymatic activities 366:394 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 5 23 theme resolution 607:616 arg1 structures 626:635 high resolution crystal structures 602:635 high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates 602:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 2 24 theme connective 293:302 arg1 disorders 311:319 severe connective tissue disorders 286:319 severe connective tissue disorders 286:319 Malfunctions in these enzymes cause severe connective tissue disorders. 30089812 9 25 theme molecular 1175:1183 arg1 mechanisms 1185:1194 the molecular mechanisms 1171:1194 the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases 1171:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 1 26 theme extracellular 196:208 arg1 matrix 210:215 extracellular matrix 196:215 extracellular matrix (ECM) maturation 196:232 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 1 26 theme extracellular 196:208 arg1 ECM 218:220 ECM 218:220 ECM 218:220 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 5 27 theme crystal 618:624 arg1 structures 626:635 high resolution crystal structures 602:635 high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates 602:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 1 28 theme Lysyl 102:106 arg1 hydroxylases 108:119 Lysyl hydroxylases 102:119 Lysyl hydroxylases 102:119 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 2 29 from Malfunctions 250:261 arg1 enzymes 272:278 these enzymes 266:278 these enzymes 266:278 Malfunctions in these enzymes cause severe connective tissue disorders. 30089812 1 30 theme matrix 210:215 arg1 maturation 223:232 extracellular matrix (ECM) maturation 196:232 extracellular matrix (ECM) maturation 196:232 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 0 31 theme Molecular 0:8 arg1 architecture 10:21 Molecular architecture 0:21 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 0:100 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 6 32 theme monomer 832:838 arg1 N- 795:796 N- 795:796 N- 795:796 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 6 32 theme monomer 832:838 arg1 boundaries 813:822 C-terminal boundaries 802:822 C-terminal boundaries 802:822 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 7 33 theme glycosyltransferase 879:897 arg1 domain 899:904 The glycosyltransferase domain 875:904 The glycosyltransferase domain 875:904 The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. 30089812 3 34 theme subsequent 458:467 arg1 glycosylation 469:481 their subsequent glycosylation 452:481 their subsequent glycosylation 452:481 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 6 35 theme accessory 857:865 arg1 domain 867:872 an accessory domain 854:872 an accessory domain 854:872 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 6 36 theme LH3 736:738 arg1 architecture 740:751 The elongated homodimeric LH3 architecture 710:751 The elongated homodimeric LH3 architecture 710:751 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 8 37 theme disease-related 991:1005 arg1 mutations 1007:1015 Known disease-related mutations 985:1015 Known disease-related mutations 985:1015 Known disease-related mutations map in close proximity to the catalytic sites. 30089812 6 38 theme homodimeric 724:734 arg1 architecture 740:751 The elongated homodimeric LH3 architecture 710:751 The elongated homodimeric LH3 architecture 710:751 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 8 39 theme Known 985:989 arg1 mutations 1007:1015 Known disease-related mutations 985:1015 Known disease-related mutations 985:1015 Known disease-related mutations map in close proximity to the catalytic sites. 30089812 0 40 theme collagen 46:53 arg1 hydroxylase 61:71 the multifunctional collagen lysyl hydroxylase 26:71 the multifunctional collagen lysyl hydroxylase 26:71 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 6 41 theme elongated 714:722 arg1 architecture 740:751 The elongated homodimeric LH3 architecture 710:751 The elongated homodimeric LH3 architecture 710:751 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 0 42 theme multifunctional 30:44 arg1 hydroxylase 61:71 the multifunctional collagen lysyl hydroxylase 26:71 the multifunctional collagen lysyl hydroxylase 26:71 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 4 43 theme information 572:582 arg1 lack 544:547 lack 544:547 lack of molecular structure information 544:582 Our understanding of LH3 functions is currently hampered by lack of molecular structure information. 30089812 4 44 theme LH3 505:507 arg1 functions 509:517 LH3 functions 505:517 LH3 functions 505:517 Our understanding of LH3 functions is currently hampered by lack of molecular structure information. 30089812 9 45 theme human 1235:1239 arg1 hydroxylases 1247:1258 human lysyl hydroxylases 1235:1258 human lysyl hydroxylases 1235:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 0 46 theme hydroxylase 61:71 arg1 architecture 10:21 Molecular architecture 0:21 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 0:100 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 4 47 theme structure 562:570 arg1 information 572:582 molecular structure information 552:582 molecular structure information 552:582 Our understanding of LH3 functions is currently hampered by lack of molecular structure information. 30089812 9 48 theme lysyl 1241:1245 arg1 hydroxylases 1247:1258 human lysyl hydroxylases 1235:1258 human lysyl hydroxylases 1235:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 5 49 from structures 626:635 arg1 complex 665:671 complex 665:671 complex with cofactors and donor substrates 665:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 0 50 theme lysyl 55:59 arg1 hydroxylase 61:71 the multifunctional collagen lysyl hydroxylase 26:71 the multifunctional collagen lysyl hydroxylase 26:71 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 6 51 theme C-terminal 802:811 arg1 boundaries 813:822 C-terminal boundaries 802:822 C-terminal boundaries 802:822 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 1 52 theme collagen 147:154 arg1 lysines 156:162 collagen lysines 147:162 collagen lysines 147:162 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 3 53 theme Human 322:326 arg1 hydroxylase 334:344 Human lysyl hydroxylase 3 322:346 Human lysyl hydroxylase 3 (LH3/PLOD3) 322:358 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 3 53 theme Human 322:326 arg1 LH3/PLOD3 349:357 LH3/PLOD3 349:357 LH3/PLOD3 349:357 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 2 54 theme severe 286:291 arg1 disorders 311:319 severe connective tissue disorders 286:319 severe connective tissue disorders 286:319 Malfunctions in these enzymes cause severe connective tissue disorders. 30089812 5 55 theme high 602:605 arg1 structures 626:635 high resolution crystal structures 602:635 high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates 602:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 1 56 theme lysines 156:162 arg1 hydroxylation 130:142 hydroxylation 130:142 hydroxylation of collagen lysines 130:162 Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. 30089812 3 57 theme lysyl 328:332 arg1 hydroxylase 334:344 Human lysyl hydroxylase 3 322:346 Human lysyl hydroxylase 3 (LH3/PLOD3) 322:358 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 3 57 theme lysyl 328:332 arg1 LH3/PLOD3 349:357 LH3/PLOD3 349:357 LH3/PLOD3 349:357 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 6 58 from boundaries 813:822 arg1 sites 782:786 two distinct catalytic sites 759:786 two distinct catalytic sites 759:786 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 6 59 dep N- 795:796 arg1 the 791:793 the 791:793 the 791:793 The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. 30089812 5 60 with complex 665:671 arg1 cofactors 678:686 cofactors 678:686 cofactors 678:686 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 5 60 with complex 665:671 arg1 substrates 698:707 donor substrates 692:707 donor substrates 692:707 Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. 30089812 0 61 theme glycosyltransferase 77:95 arg1 LH3 97:99 glycosyltransferase LH3 77:99 glycosyltransferase LH3 77:99 Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3. 30089812 9 62 theme diseases 1216:1223 arg1 mechanisms 1185:1194 the molecular mechanisms 1171:1194 the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases 1171:1258 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 9 62 theme diseases 1216:1223 arg1 functions 1149:1157 the multiple functions 1136:1157 the multiple functions of LH3 1136:1164 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 9 63 theme multiple 1140:1147 arg1 functions 1149:1157 the multiple functions 1136:1157 the multiple functions of LH3 1136:1164 Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases. 30089812 3 64 theme lysine 422:427 arg1 hydroxylation 429:441 collagen lysine hydroxylation 413:441 collagen lysine hydroxylation 413:441 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 3 65 contain bears 360:364 arg2 activities 385:394 multiple enzymatic activities 366:394 multiple enzymatic activities 366:394 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 3 65 contain bears 360:364 arg1 hydroxylase 334:344 Human lysyl hydroxylase 3 322:346 Human lysyl hydroxylase 3 (LH3/PLOD3) 322:358 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 3 65 contain bears 360:364 arg1 LH3/PLOD3 349:357 LH3/PLOD3 349:357 LH3/PLOD3 349:357 Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. 30089812 7 66 theme known 957:961 arg1 glycosyltransferases 963:982 other known glycosyltransferases 951:982 other known glycosyltransferases 951:982 The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. 15477100 1 0 theme most 146:149 arg1 fluids 158:163 most bodily fluids 146:163 most bodily fluids 146:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 1 theme bodily 151:156 arg1 fluids 158:163 most bodily fluids 146:163 most bodily fluids 146:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 2 2 attach isolated 221:228 arg1 serum 241:245 human serum 235:245 human serum 235:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 2 2 attach isolated 221:228 arg2 ZAG 217:219 ZAG 217:219 ZAG isolated from human serum 217:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 5 3 theme ZAG 934:936 arg1 groove 938:943 the ZAG groove 930:943 the ZAG groove 930:943 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 2 4 theme MHC 344:346 arg1 molecules 349:357 ZAG and class I major histocompatibility complex (MHC) molecules 294:357 molecules 349:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 5 from ligand 739:744 arg1 structure 783:791 the structure 779:791 the structure of serum ZAG 779:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 5 from ligand 739:744 arg1 structures 761:770 the current structures 749:770 the current structures 749:770 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 3 6 theme non-peptidic 530:541 arg1 ligand 543:548 the non-peptidic ligand 526:548 the non-peptidic ligand 526:548 Here we present crystallographic studies to explore further the nature of the non-peptidic ligand in the ZAG groove. 15477100 2 7 theme ZAG 294:296 arg1 molecules 349:357 ZAG and class I major histocompatibility complex (MHC) molecules 294:357 molecules 349:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 8 theme non-peptidic 726:737 arg1 ligand 739:744 the non-peptidic ligand 722:744 the non-peptidic ligand in the current structures and in the structure of serum ZAG 722:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 8 theme non-peptidic 726:737 arg1 present 847:853 present 847:853 present 847:853 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 8 theme non-peptidic 726:737 arg1 glycol 824:829 a polyethylene glycol 809:829 a polyethylene glycol (PEG) 809:835 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 1 9 from present 135:141 arg1 fluids 158:163 most bodily fluids 146:163 most bodily fluids 146:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 3 10 theme ligand 543:548 arg1 nature 516:521 the nature 512:521 the nature of the non-peptidic ligand in the ZAG groove 512:566 Here we present crystallographic studies to explore further the nature of the non-peptidic ligand in the ZAG groove. 15477100 3 11 theme crystallographic 468:483 arg1 studies 485:491 crystallographic studies 468:491 crystallographic studies 468:491 Here we present crystallographic studies to explore further the nature of the non-peptidic ligand in the ZAG groove. 15477100 1 12 attach present 135:141 arg2 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein (ZAG) 70:97 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 12 attach present 135:141 arg1 fluids 158:163 most bodily fluids 146:163 most bodily fluids 146:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 12 attach present 135:141 arg2 present 135:141 present 135:141 present 135:141 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 12 attach present 135:141 arg2 protein 119:125 a 41 kDa soluble protein 102:125 a 41 kDa soluble protein that is present in most bodily fluids 102:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 6 13 theme ZAG 1170:1172 arg1 capable 1184:1190 capable 1184:1190 capable 1184:1190 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 6 13 theme ZAG 1170:1172 arg1 groove 1174:1179 the ZAG groove 1166:1179 the ZAG groove 1166:1179 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 3 14 from nature 516:521 arg1 groove 561:566 the ZAG groove 553:566 the ZAG groove 553:566 Here we present crystallographic studies to explore further the nature of the non-peptidic ligand in the ZAG groove. 15477100 2 15 theme structural 264:273 arg1 similarity 275:284 the structural similarity 260:284 the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules 260:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 16 located present 847:853 arg2 ligand 739:744 the non-peptidic ligand 722:744 the non-peptidic ligand in the current structures and in the structure of serum ZAG 722:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 16 located present 847:853 arg2 PEG 832:834 PEG 832:834 PEG 832:834 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 16 located present 847:853 arg2 present 847:853 present 847:853 present 847:853 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 16 located present 847:853 arg2 glycol 824:829 a polyethylene glycol 809:829 a polyethylene glycol (PEG) 809:835 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 16 located present 847:853 arg1 conditions 878:887 the crystallization conditions 858:887 the crystallization conditions used 858:892 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 5 17 theme binding 1036:1042 arg1 site 1044:1047 the ZAG binding site 1028:1047 the ZAG binding site 1028:1047 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 4 18 theme ZAG 802:804 arg1 structure 783:791 the structure 779:791 the structure of serum ZAG 779:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 0 19 theme Crystallographic 0:15 arg1 studies 17:23 Crystallographic studies 0:23 Crystallographic studies of ligand binding by Zn-alpha2-glycoprotein 0:67 Crystallographic studies of ligand binding by Zn-alpha2-glycoprotein. 15477100 5 20 theme ZAG 1032:1034 arg1 site 1044:1047 the ZAG binding site 1028:1047 the ZAG binding site 1028:1047 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 2 21 theme non-peptidic 374:385 arg1 ligand 387:392 a non-peptidic ligand 372:392 a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove 372:449 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 2 22 theme peptide-binding 428:442 arg1 groove 444:449 the MHC peptide-binding groove 420:449 the MHC peptide-binding groove 420:449 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 23 theme polyethylene 811:822 arg1 ligand 739:744 the non-peptidic ligand 722:744 the non-peptidic ligand in the current structures and in the structure of serum ZAG 722:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 23 theme polyethylene 811:822 arg1 present 847:853 present 847:853 present 847:853 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 23 theme polyethylene 811:822 arg1 PEG 832:834 PEG 832:834 PEG 832:834 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 23 theme polyethylene 811:822 arg1 glycol 824:829 a polyethylene glycol 809:829 a polyethylene glycol (PEG) 809:835 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 3 24 theme ZAG 557:559 arg1 groove 561:566 the ZAG groove 553:566 the ZAG groove 553:566 Here we present crystallographic studies to explore further the nature of the non-peptidic ligand in the ZAG groove. 15477100 1 25 located present 135:141 arg2 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein (ZAG) 70:97 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 25 located present 135:141 arg1 fluids 158:163 most bodily fluids 146:163 most bodily fluids 146:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 25 located present 135:141 arg2 present 135:141 present 135:141 present 135:141 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 25 located present 135:141 arg2 protein 119:125 a 41 kDa soluble protein 102:125 a 41 kDa soluble protein that is present in most bodily fluids 102:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 2 26 theme MHC 424:426 arg1 groove 444:449 the MHC peptide-binding groove 420:449 the MHC peptide-binding groove 420:449 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 27 theme insect 694:699 arg1 cells 701:705 insect cells 694:705 insect cells 694:705 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 2 28 theme class 302:306 arg1 molecules 349:357 ZAG and class I major histocompatibility complex (MHC) molecules 294:357 molecules 349:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 0 29 theme ligand 28:33 arg1 binding 35:41 ligand binding 28:41 ligand binding by Zn-alpha2-glycoprotein 28:67 Crystallographic studies of ligand binding by Zn-alpha2-glycoprotein. 15477100 2 30 theme human 235:239 arg1 serum 241:245 human serum 235:245 human serum 235:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 31 from present 847:853 arg1 conditions 878:887 the crystallization conditions 858:887 the crystallization conditions used 858:892 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 32 theme recombinant 618:628 arg1 ZAG 630:632 recombinant ZAG 618:632 recombinant ZAG 618:632 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 6 33 theme ZAG 1111:1113 arg1 forms 1102:1106 our purified forms 1089:1106 our purified forms of ZAG 1089:1113 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 4 34 from conditions 878:887 arg1 ligand 739:744 the non-peptidic ligand 722:744 the non-peptidic ligand in the current structures and in the structure of serum ZAG 722:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 34 from conditions 878:887 arg1 present 847:853 present 847:853 present 847:853 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 34 from conditions 878:887 arg1 glycol 824:829 a polyethylene glycol 809:829 a polyethylene glycol (PEG) 809:835 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 1 35 theme kDa 107:109 arg1 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein (ZAG) 70:97 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 35 theme kDa 107:109 arg1 protein 119:125 a 41 kDa soluble protein 102:125 a 41 kDa soluble protein that is present in most bodily fluids 102:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 35 theme kDa 107:109 arg1 present 135:141 present 135:141 present 135:141 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 4 36 attach derived 664:670 arg2 structure 654:662 A structure 652:662 a 1.95 A structure derived from ZAG expressed in insect cells 645:705 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 36 attach derived 664:670 arg1 ZAG 677:679 ZAG 677:679 ZAG expressed in insect cells 677:705 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 2 37 theme ZAG 401:403 arg1 counterpart 405:415 the ZAG counterpart 397:415 the ZAG counterpart of the MHC peptide-binding groove 397:449 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 6 38 theme endogenous 1138:1147 arg1 ligand 1149:1154 a bound endogenous ligand 1130:1154 a bound endogenous ligand 1130:1154 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 5 39 theme fatty 1012:1016 arg1 acid 1018:1021 a fluorophore-tagged fatty acid 991:1021 a fluorophore-tagged fatty acid 991:1021 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 4 40 attach present 847:853 arg2 ligand 739:744 the non-peptidic ligand 722:744 the non-peptidic ligand in the current structures and in the structure of serum ZAG 722:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 40 attach present 847:853 arg2 PEG 832:834 PEG 832:834 PEG 832:834 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 40 attach present 847:853 arg2 present 847:853 present 847:853 present 847:853 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 40 attach present 847:853 arg2 glycol 824:829 a polyethylene glycol 809:829 a polyethylene glycol (PEG) 809:835 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 40 attach present 847:853 arg1 conditions 878:887 the crystallization conditions 858:887 the crystallization conditions used 858:892 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 0 41 theme binding 35:41 arg1 studies 17:23 Crystallographic studies 0:23 Crystallographic studies of ligand binding by Zn-alpha2-glycoprotein 0:67 Crystallographic studies of ligand binding by Zn-alpha2-glycoprotein. 15477100 4 42 theme crystallization 862:876 arg1 conditions 878:887 the crystallization conditions 858:887 the crystallization conditions used 858:892 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 2 43 theme ZAG 217:219 arg1 structure 204:212 The previously reported 2.8 A crystal structure 166:212 The previously reported 2.8 A crystal structure of ZAG isolated from human serum 166:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 44 theme ZAG 630:632 arg1 structure 654:662 A structure 652:662 a 1.95 A structure derived from ZAG expressed in insect cells 645:705 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 44 theme ZAG 630:632 arg1 forms 609:613 several forms 601:613 several forms 601:613 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 5 45 theme fluorophore-tagged 993:1010 arg1 acid 1018:1021 a fluorophore-tagged fatty acid 991:1021 a fluorophore-tagged fatty acid 991:1021 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 4 46 theme serum 796:800 arg1 ZAG 802:804 serum ZAG 796:804 serum ZAG 796:804 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 1 47 theme soluble 111:117 arg1 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein (ZAG) 70:97 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 47 theme soluble 111:117 arg1 protein 119:125 a 41 kDa soluble protein 102:125 a 41 kDa soluble protein that is present in most bodily fluids 102:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 47 theme soluble 111:117 arg1 present 135:141 present 135:141 present 135:141 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 2 48 theme groove 444:449 arg1 counterpart 405:415 the ZAG counterpart 397:415 the ZAG counterpart of the MHC peptide-binding groove 397:449 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 6 49 contain contain 1122:1128 arg1 forms 1102:1106 our purified forms 1089:1106 our purified forms of ZAG 1089:1113 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 6 49 contain contain 1122:1128 arg2 ligand 1149:1154 a bound endogenous ligand 1130:1154 a bound endogenous ligand 1130:1154 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 2 50 theme crystal 196:202 arg1 structure 204:212 The previously reported 2.8 A crystal structure 166:212 The previously reported 2.8 A crystal structure of ZAG isolated from human serum 166:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 6 51 theme bound 1132:1136 arg1 ligand 1149:1154 a bound endogenous ligand 1130:1154 a bound endogenous ligand 1130:1154 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 2 52 from ligand 387:392 arg1 counterpart 405:415 the ZAG counterpart 397:415 the ZAG counterpart of the MHC peptide-binding groove 397:449 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 2 53 theme complex 335:341 arg1 molecules 349:357 ZAG and class I major histocompatibility complex (MHC) molecules 294:357 molecules 349:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 6 54 theme hydrophobic 1203:1213 arg1 molecules 1215:1223 hydrophobic molecules 1203:1223 hydrophobic molecules 1203:1223 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 2 55 theme A 194:194 arg1 structure 204:212 The previously reported 2.8 A crystal structure 166:212 The previously reported 2.8 A crystal structure of ZAG isolated from human serum 166:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 2 56 theme histocompatibility 316:333 arg1 molecules 349:357 ZAG and class I major histocompatibility complex (MHC) molecules 294:357 molecules 349:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 5 57 theme Further 895:901 arg1 support 903:909 Further support 895:909 Further support for PEG binding in the ZAG groove 895:943 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 6 58 theme purified 1093:1100 arg1 forms 1102:1106 our purified forms 1089:1106 our purified forms of ZAG 1089:1113 From these results we hypothesize that our purified forms of ZAG do not contain a bound endogenous ligand, but that the ZAG groove is capable of binding hydrophobic molecules, which may relate to its function. 15477100 2 59 theme 2.8 190:192 arg1 structure 204:212 The previously reported 2.8 A crystal structure 166:212 The previously reported 2.8 A crystal structure of ZAG isolated from human serum 166:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 60 theme current 753:759 arg1 structures 761:770 the current structures 749:770 the current structures 749:770 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 2 61 theme major 310:314 arg1 molecules 349:357 ZAG and class I major histocompatibility complex (MHC) molecules 294:357 molecules 349:357 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 4 62 theme structures 587:596 arg1 Comparison 569:578 Comparison 569:578 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, 569:706 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 2 63 theme reported 181:188 arg1 structure 204:212 The previously reported 2.8 A crystal structure 166:212 The previously reported 2.8 A crystal structure of ZAG isolated from human serum 166:245 The previously reported 2.8 A crystal structure of ZAG isolated from human serum demonstrated the structural similarity between ZAG and class I major histocompatibility complex (MHC) molecules and revealed a non-peptidic ligand in the ZAG counterpart of the MHC peptide-binding groove. 15477100 0 64 gly Zn-alpha2-glycoprotein 46:67 arg1 Zn-alpha2-glycoprotein 46:67 Zn-alpha2-glycoprotein 46:67 Zn-alpha2-glycoprotein 46:67 Crystallographic studies of ligand binding by Zn-alpha2-glycoprotein. 15477100 5 65 from support 903:909 arg1 groove 938:943 the ZAG groove 930:943 the ZAG groove 930:943 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 4 66 theme forms 609:613 arg1 structures 587:596 the structures 583:596 the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, 583:706 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 1 67 gly Zn-alpha2-glycoprotein 70:91 arg1 ZAG 94:96 ZAG 94:96 ZAG 94:96 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 67 gly Zn-alpha2-glycoprotein 70:91 arg1 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein (ZAG) 70:97 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 67 gly Zn-alpha2-glycoprotein 70:91 arg1 protein 119:125 a 41 kDa soluble protein 102:125 a 41 kDa soluble protein that is present in most bodily fluids 102:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 67 gly Zn-alpha2-glycoprotein 70:91 arg1 present 135:141 present 135:141 present 135:141 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 5 68 theme PEG 915:917 arg1 binding 919:925 PEG binding 915:925 PEG binding 915:925 Further support for PEG binding in the ZAG groove is provided by the finding that PEG displaces a fluorophore-tagged fatty acid from the ZAG binding site. 15477100 1 69 from fluids 158:163 arg1 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein 70:91 Zn-alpha2-glycoprotein (ZAG) 70:97 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 69 from fluids 158:163 arg1 protein 119:125 a 41 kDa soluble protein 102:125 a 41 kDa soluble protein that is present in most bodily fluids 102:163 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 1 69 from fluids 158:163 arg1 present 135:141 present 135:141 present 135:141 Zn-alpha2-glycoprotein (ZAG) is a 41 kDa soluble protein that is present in most bodily fluids. 15477100 4 70 theme several 601:607 arg1 structure 654:662 A structure 652:662 a 1.95 A structure derived from ZAG expressed in insect cells 645:705 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 15477100 4 70 theme several 601:607 arg1 forms 609:613 several forms 601:613 several forms 601:613 Comparison of the structures of several forms of recombinant ZAG, including a 1.95 A structure derived from ZAG expressed in insect cells, suggests that the non-peptidic ligand in the current structures and in the structure of serum ZAG is a polyethylene glycol (PEG), which is present in the crystallization conditions used. 3123586 5 0 theme acid 697:700 arg1 replacement 702:712 this amino acid replacement 686:712 this amino acid replacement 686:712 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 1 1 theme major 136:140 arg1 C-III 114:118 Apolipoprotein C-III 99:118 Apolipoprotein C-III (apoC-III) 99:129 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 1 theme major 136:140 arg1 protein 142:148 a major protein 134:148 a major protein of very low density lipoprotein (VLDL) 134:187 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 2 2 contain contains 215:222 arg2 chain 239:243 a carbohydrate chain 224:243 a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74 224:356 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 2 contain contains 215:222 arg1 polypeptide 203:213 The apoC-III polypeptide 190:213 The apoC-III polypeptide 190:213 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 5 3 theme replacement 702:712 arg1 result 676:681 a result 674:681 a result of this amino acid replacement 674:712 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 5 3 theme replacement 702:712 arg1 polypeptide 735:745 the mutant apoC-III polypeptide 715:745 the mutant apoC-III polypeptide 715:745 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 4 4 from position 626:633 arg1 alanine 615:621 an alanine 612:621 an alanine at position 74 instead of the normal threonine 612:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 4 from position 626:633 arg1 substitution 577:588 a single nucleotide substitution 557:588 a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine 557:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 6 5 theme genomic 889:895 arg1 DNA 897:899 genomic DNA 889:899 genomic DNA 889:899 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 2 6 theme threonine 325:333 arg1 residue 335:341 a threonine residue 323:341 a threonine residue at position 74 323:356 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 4 7 theme gene 543:546 arg1 analysis 520:527 DNA sequence analysis 507:527 DNA sequence analysis of the cloned gene 507:546 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 6 8 theme AluI 818:821 arg1 site 823:826 a novel AluI site 810:826 a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA 810:899 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 5 9 theme apoC-III 726:733 arg1 result 676:681 a result 674:681 a result of this amino acid replacement 674:712 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 5 9 theme apoC-III 726:733 arg1 polypeptide 735:745 the mutant apoC-III polypeptide 715:745 the mutant apoC-III polypeptide 715:745 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 2 10 attach attached 298:305 arg3 O-linkage 310:318 O-linkage 310:318 O-linkage 310:318 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 10 attach attached 298:305 arg1 residue 335:341 a threonine residue 323:341 a threonine residue at position 74 323:356 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 10 attach attached 298:305 arg2 galactosamine 256:268 galactosamine 256:268 galactosamine 256:268 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 4 11 theme cloned 536:541 arg1 gene 543:546 the cloned gene 532:546 the cloned gene 532:546 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 12 theme sequence 511:518 arg1 analysis 520:527 DNA sequence analysis 507:527 DNA sequence analysis of the cloned gene 507:546 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 1 13 theme low 158:160 arg1 VLDL 183:186 VLDL 183:186 VLDL 183:186 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 13 theme low 158:160 arg1 lipoprotein 170:180 very low density lipoprotein 153:180 very low density lipoprotein (VLDL) 153:187 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 0 14 theme Molecular 0:8 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a human apoC-III variant 0:44 Molecular cloning of a human apoC-III variant: Thr 74----Ala 74 mutation prevents O-glycosylation. 3123586 2 15 contain containing 245:254 arg2 acid 293:296 sialic acid 286:296 sialic acid 286:296 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 15 contain containing 245:254 arg2 galactose 271:279 galactose 271:279 galactose 271:279 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 15 contain containing 245:254 arg2 galactosamine 256:268 galactosamine 256:268 galactosamine 256:268 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 15 contain containing 245:254 arg1 chain 239:243 a carbohydrate chain 224:243 a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74 224:356 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 2 16 theme sialic 286:291 arg1 acid 293:296 sialic acid 286:296 sialic acid 286:296 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 1 17 theme density 162:168 arg1 VLDL 183:186 VLDL 183:186 VLDL 183:186 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 17 theme density 162:168 arg1 lipoprotein 170:180 very low density lipoprotein 153:180 very low density lipoprotein (VLDL) 153:187 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 4 18 theme DNA 507:509 arg1 analysis 520:527 DNA sequence analysis 507:527 DNA sequence analysis of the cloned gene 507:546 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 1 19 theme lipoprotein 170:180 arg1 C-III 114:118 Apolipoprotein C-III 99:118 Apolipoprotein C-III (apoC-III) 99:129 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 19 theme lipoprotein 170:180 arg1 protein 142:148 a major protein 134:148 a major protein of very low density lipoprotein (VLDL) 134:187 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 20 theme Apolipoprotein 99:112 arg1 apoC-III 121:128 apoC-III 121:128 apoC-III 121:128 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 20 theme Apolipoprotein 99:112 arg1 C-III 114:118 Apolipoprotein C-III 99:118 Apolipoprotein C-III (apoC-III) 99:129 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 1 20 theme Apolipoprotein 99:112 arg1 protein 142:148 a major protein 134:148 a major protein of very low density lipoprotein (VLDL) 134:187 Apolipoprotein C-III (apoC-III) is a major protein of very low density lipoprotein (VLDL). 3123586 0 21 theme apoC-III 29:36 arg1 variant 38:44 a human apoC-III variant 21:44 a human apoC-III variant 21:44 Molecular cloning of a human apoC-III variant: Thr 74----Ala 74 mutation prevents O-glycosylation. 3123586 6 22 theme novel 812:816 arg1 site 823:826 a novel AluI site 810:826 a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA 810:899 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 0 23 theme human 23:27 arg1 variant 38:44 a human apoC-III variant 21:44 a human apoC-III variant 21:44 Molecular cloning of a human apoC-III variant: Thr 74----Ala 74 mutation prevents O-glycosylation. 3123586 6 24 theme Southern 868:875 arg1 blotting 877:884 Southern blotting 868:884 Southern blotting of genomic DNA 868:899 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 3 25 theme apoC-III 378:385 arg1 gene 387:390 the apoC-III gene 374:390 the apoC-III gene 374:390 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 6 26 theme apoC-III 788:795 arg1 gene 797:800 the apoC-III gene 784:800 the apoC-III gene 784:800 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 2 27 theme apoC-III 194:201 arg1 polypeptide 203:213 The apoC-III polypeptide 190:213 The apoC-III polypeptide 190:213 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 6 28 from mutation 772:779 arg1 gene 797:800 the apoC-III gene 784:800 the apoC-III gene 784:800 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 5 29 theme mutant 719:724 arg1 result 676:681 a result 674:681 a result of this amino acid replacement 674:712 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 5 29 theme mutant 719:724 arg1 polypeptide 735:745 the mutant apoC-III polypeptide 715:745 the mutant apoC-III polypeptide 715:745 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 3 30 contain contained 419:427 arg1 serum 413:417 serum 413:417 serum 413:417 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 3 30 contain contained 419:427 arg2 apoC-III 455:462 apoC-III 455:462 apoC-III lacking the carbohydrate moiety (C-III-0) 455:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 3 30 contain contained 419:427 arg2 amounts 444:450 unusually high amounts 429:450 unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0) 429:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 5 31 gly glycosylated 754:765 arg1 result 676:681 a result 674:681 a result of this amino acid replacement 674:712 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 5 31 gly glycosylated 754:765 arg1 polypeptide 735:745 the mutant apoC-III polypeptide 715:745 the mutant apoC-III polypeptide 715:745 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 2 32 from position 346:353 arg1 residue 335:341 a threonine residue 323:341 a threonine residue at position 74 323:356 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 4 33 theme normal 653:658 arg1 threonine 660:668 the normal threonine 649:668 an alanine at position 74 instead of the normal threonine 612:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 33 theme normal 653:658 arg1 substitution 577:588 a single nucleotide substitution 557:588 a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine 557:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 0 34 theme variant 38:44 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a human apoC-III variant 0:44 Molecular cloning of a human apoC-III variant: Thr 74----Ala 74 mutation prevents O-glycosylation. 3123586 6 35 theme change 858:863 arg1 diagnosis 841:849 diagnosis 841:849 diagnosis of the change 841:863 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 3 36 theme apoC-III 455:462 arg1 apoC-III 455:462 apoC-III 455:462 apoC-III lacking the carbohydrate moiety (C-III-0) 455:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 3 36 theme apoC-III 455:462 arg1 amounts 444:450 unusually high amounts 429:450 unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0) 429:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 5 37 theme amino 691:695 arg1 replacement 702:712 this amino acid replacement 686:712 this amino acid replacement 686:712 As a result of this amino acid replacement, the mutant apoC-III polypeptide is not glycosylated. 3123586 3 38 theme high 439:442 arg1 apoC-III 455:462 apoC-III 455:462 apoC-III lacking the carbohydrate moiety (C-III-0) 455:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 3 38 theme high 439:442 arg1 amounts 444:450 unusually high amounts 429:450 unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0) 429:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 4 39 theme single 559:564 arg1 threonine 660:668 the normal threonine 649:668 an alanine at position 74 instead of the normal threonine 612:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 39 theme single 559:564 arg1 alanine 615:621 an alanine 612:621 an alanine at position 74 instead of the normal threonine 612:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 39 theme single 559:564 arg1 A----G 591:596 A----G 591:596 A----G 591:596 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 39 theme single 559:564 arg1 substitution 577:588 a single nucleotide substitution 557:588 a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine 557:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 2 40 theme carbohydrate 226:237 arg1 chain 239:243 a carbohydrate chain 224:243 a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74 224:356 The apoC-III polypeptide contains a carbohydrate chain containing galactosamine, galactose, and sialic acid attached in O-linkage to a threonine residue at position 74. 3123586 0 41 theme 74----Ala 51:59 arg1 mutation 64:71 Thr 74----Ala 74 mutation 47:71 Thr 74----Ala 74 mutation 47:71 Molecular cloning of a human apoC-III variant: Thr 74----Ala 74 mutation prevents O-glycosylation. 3123586 3 42 theme carbohydrate 476:487 arg1 C-III-0 497:503 C-III-0 497:503 C-III-0 497:503 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 3 42 theme carbohydrate 476:487 arg1 moiety 489:494 the carbohydrate moiety 472:494 the carbohydrate moiety (C-III-0) 472:504 We have cloned the apoC-III gene from a subject whose serum contained unusually high amounts of apoC-III lacking the carbohydrate moiety (C-III-0). 3123586 0 43 theme Thr 47:49 arg1 mutation 64:71 Thr 74----Ala 74 mutation 47:71 Thr 74----Ala 74 mutation 47:71 Molecular cloning of a human apoC-III variant: Thr 74----Ala 74 mutation prevents O-glycosylation. 3123586 6 44 theme DNA 897:899 arg1 blotting 877:884 Southern blotting 868:884 Southern blotting of genomic DNA 868:899 The mutation in the apoC-III gene creates a novel AluI site that permits diagnosis of the change by Southern blotting of genomic DNA. 3123586 4 45 theme nucleotide 566:575 arg1 threonine 660:668 the normal threonine 649:668 an alanine at position 74 instead of the normal threonine 612:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 45 theme nucleotide 566:575 arg1 alanine 615:621 an alanine 612:621 an alanine at position 74 instead of the normal threonine 612:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 45 theme nucleotide 566:575 arg1 A----G 591:596 A----G 591:596 A----G 591:596 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 3123586 4 45 theme nucleotide 566:575 arg1 substitution 577:588 a single nucleotide substitution 557:588 a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine 557:668 DNA sequence analysis of the cloned gene revealed a single nucleotide substitution (A----G) that encodes an alanine at position 74 instead of the normal threonine. 19683538 5 0 theme alpha-NAGAL 867:877 arg1 enzyme 879:884 the human alpha-NAGAL enzyme 857:884 the human alpha-NAGAL enzyme 857:884 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 6 1 from effect 1127:1132 arg1 structure 1188:1196 the three-dimensional structure 1166:1196 the three-dimensional structure 1166:1196 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 3 2 gly glycoproteins 605:617 arg1 glycoproteins 605:617 wild-type and glycosylation-deficient glycoproteins 567:617 wild-type and glycosylation-deficient glycoproteins 567:617 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 5 3 theme enzyme 879:884 arg1 mechanism 844:852 catalytic mechanism 834:852 catalytic mechanism 834:852 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 5 3 theme enzyme 879:884 arg1 specificity 818:828 binding specificity 810:828 binding specificity 810:828 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 5 4 dep different 939:947 arg1 catalytic 949:957 catalytic 949:957 catalytic 949:957 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 2 5 theme Schindler 394:402 arg1 disease 404:410 Schindler disease 394:410 Schindler disease 394:410 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 5 theme Schindler 394:402 arg1 disorders 384:392 the lysosomal storage disorders 362:392 the lysosomal storage disorders Schindler disease and Kanzaki disease 362:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 3 6 from defects 468:474 arg1 diseases 483:490 the diseases 479:490 the diseases 479:490 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 3 7 theme wild-type 567:575 arg1 glycoproteins 605:617 wild-type and glycosylation-deficient glycoproteins 567:617 wild-type and glycosylation-deficient glycoproteins 567:617 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 3 8 theme cell 641:644 arg1 systems 657:663 recombinant insect cell expression systems 622:663 recombinant insect cell expression systems 622:663 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 0 9 theme Schindler 85:93 arg1 basis 76:80 The molecular basis 62:80 The molecular basis of Schindler and Kanzaki diseases 62:114 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 4 10 theme purified 710:717 arg1 enzymes 740:746 our purified wild-type and mutant enzymes 706:746 our purified wild-type and mutant enzymes 706:746 We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. 19683538 3 11 theme recombinant 622:632 arg1 systems 657:663 recombinant insect cell expression systems 622:663 recombinant insect cell expression systems 622:663 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 1 12 dep alpha-NAGAL 150:160 arg1 E.C. 163:166 E.C. 163:166 E.C. 163:166 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 1 13 gly glycopeptides 272:284 arg2 glycopeptides 272:284 glycopeptides 272:284 glycopeptides 272:284 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 3 14 theme glycosylation-deficient 581:603 arg1 glycoproteins 605:617 wild-type and glycosylation-deficient glycoproteins 567:617 wild-type and glycosylation-deficient glycoproteins 567:617 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 0 15 theme Kanzaki 99:105 arg1 basis 76:80 The molecular basis 62:80 The molecular basis of Schindler and Kanzaki diseases 62:114 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 3 16 theme molecular 458:466 arg1 defects 468:474 the molecular defects 454:474 the molecular defects in the diseases 454:490 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 0 17 dep Schindler 85:93 arg1 diseases 107:114 diseases 107:114 diseases 107:114 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 4 18 theme enzymes 740:746 arg1 parameters 692:701 the enzymatic parameters 678:701 the enzymatic parameters of our purified wild-type and mutant enzymes 678:746 We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. 19683538 5 19 theme crystallographic 907:922 arg1 complexes 924:932 three crystallographic complexes 901:932 three crystallographic complexes with different catalytic products bound in the active site of the enzyme 901:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 3 20 theme expression 646:655 arg1 systems 657:663 recombinant insect cell expression systems 622:663 recombinant insect cell expression systems 622:663 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 1 21 theme lysosomal 183:191 arg1 exoglycosidase 193:206 a lysosomal exoglycosidase 181:206 a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids 181:300 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 1 21 theme lysosomal 183:191 arg1 alpha-N-acetylgalactosaminidase 117:147 alpha-N-acetylgalactosaminidase 117:147 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) 117:176 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 5 22 theme binding 810:816 arg1 specificity 818:828 binding specificity 810:828 binding specificity 810:828 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 6 23 theme Schindler 1092:1100 arg1 disease 1102:1108 Schindler disease 1092:1108 Schindler disease 1092:1108 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 6 24 gly glycoprotein 1071:1082 arg1 glycoprotein 1071:1082 the alpha-NAGAL glycoprotein 1055:1082 the alpha-NAGAL glycoprotein 1055:1082 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 0 25 dep basis 76:80 arg1 structure 10:18 The 1.9 a structure 0:18 The 1.9 a structure of human alpha-N-acetylgalactosaminidase 0:59 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 5 26 with complexes 924:932 arg1 products 959:966 different catalytic products 939:966 different catalytic products bound in the active site of the enzyme 939:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 2 27 dep disorders 384:392 arg1 disease 404:410 Schindler disease 394:410 Schindler disease 394:410 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 27 dep disorders 384:392 arg1 disease 424:430 Kanzaki disease 416:430 Kanzaki disease 416:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 27 dep disorders 384:392 arg1 disorders 384:392 the lysosomal storage disorders 362:392 the lysosomal storage disorders Schindler disease and Kanzaki disease 362:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 4 28 theme wild-type 719:727 arg1 enzymes 740:746 our purified wild-type and mutant enzymes 706:746 our purified wild-type and mutant enzymes 706:746 We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. 19683538 0 29 theme a 8:8 arg1 structure 10:18 The 1.9 a structure 0:18 The 1.9 a structure of human alpha-N-acetylgalactosaminidase 0:59 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 2 30 theme alpha-NAGAL 330:340 arg1 activity 342:349 alpha-NAGAL activity 330:349 alpha-NAGAL activity 330:349 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 3 31 theme crystal 511:517 arg1 structure 519:527 the crystal structure 507:527 the crystal structure of human alpha-NAGAL 507:548 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 0 32 theme human 23:27 arg1 alpha-N-acetylgalactosaminidase 29:59 human alpha-N-acetylgalactosaminidase 23:59 human alpha-N-acetylgalactosaminidase 23:59 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 3 33 theme alpha-NAGAL 538:548 arg1 structure 519:527 the crystal structure 507:527 the crystal structure of human alpha-NAGAL 507:548 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 1 34 from glycolipids 290:300 arg1 residues 258:265 terminal alpha-N-acetylgalactosamine residues 221:265 terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids 221:300 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 4 35 theme mutant 733:738 arg1 enzymes 740:746 our purified wild-type and mutant enzymes 706:746 our purified wild-type and mutant enzymes 706:746 We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. 19683538 1 36 theme terminal 221:228 arg1 residues 258:265 terminal alpha-N-acetylgalactosamine residues 221:265 terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids 221:300 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 6 37 theme alpha-NAGAL 1059:1069 arg1 glycoprotein 1071:1082 the alpha-NAGAL glycoprotein 1055:1082 the alpha-NAGAL glycoprotein 1055:1082 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 1 38 theme alpha-N-acetylgalactosamine 230:256 arg1 residues 258:265 terminal alpha-N-acetylgalactosamine residues 221:265 terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids 221:300 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 1 39 from glycopeptides 272:284 arg1 residues 258:265 terminal alpha-N-acetylgalactosamine residues 221:265 terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids 221:300 alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. 19683538 5 40 dep specificity 818:828 arg1 the 806:808 the 806:808 the 806:808 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 3 41 theme human 532:536 arg1 alpha-NAGAL 538:548 human alpha-NAGAL 532:548 human alpha-NAGAL 532:548 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 0 42 theme alpha-N-acetylgalactosaminidase 29:59 arg1 structure 10:18 The 1.9 a structure 0:18 The 1.9 a structure of human alpha-N-acetylgalactosaminidase 0:59 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 3 43 theme insect 634:639 arg1 systems 657:663 recombinant insect cell expression systems 622:663 recombinant insect cell expression systems 622:663 To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. 19683538 6 44 from defects 1044:1050 arg1 glycoprotein 1071:1082 the alpha-NAGAL glycoprotein 1055:1082 the alpha-NAGAL glycoprotein 1055:1082 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 2 45 theme storage 376:382 arg1 disease 404:410 Schindler disease 394:410 Schindler disease 394:410 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 45 theme storage 376:382 arg1 disease 424:430 Kanzaki disease 416:430 Kanzaki disease 416:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 45 theme storage 376:382 arg1 disorders 384:392 the lysosomal storage disorders 362:392 the lysosomal storage disorders Schindler disease and Kanzaki disease 362:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 46 theme lysosomal 366:374 arg1 disease 404:410 Schindler disease 394:410 Schindler disease 394:410 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 46 theme lysosomal 366:374 arg1 disease 424:430 Kanzaki disease 416:430 Kanzaki disease 416:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 46 theme lysosomal 366:374 arg1 disorders 384:392 the lysosomal storage disorders 362:392 the lysosomal storage disorders Schindler disease and Kanzaki disease 362:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 47 theme activity 342:349 arg1 deficiency 316:325 a deficiency 314:325 a deficiency of alpha-NAGAL activity 314:349 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 6 48 theme disease-causing 1137:1151 arg1 mutations 1153:1161 disease-causing mutations 1137:1161 disease-causing mutations 1137:1161 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 5 49 theme enzyme 1000:1005 arg1 site 988:991 the active site 977:991 the active site of the enzyme 977:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 5 49 theme enzyme 1000:1005 arg1 enzyme 1000:1005 the enzyme 996:1005 the enzyme 996:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 4 50 theme enzymatic 682:690 arg1 parameters 692:701 the enzymatic parameters 678:701 the enzymatic parameters of our purified wild-type and mutant enzymes 678:746 We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. 19683538 0 51 theme molecular 66:74 arg1 basis 76:80 The molecular basis 62:80 The molecular basis of Schindler and Kanzaki diseases 62:114 The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. 19683538 6 52 theme mutations 1153:1161 arg1 effect 1127:1132 the effect 1123:1132 the effect of disease-causing mutations on the three-dimensional structure 1123:1196 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 6 53 theme three-dimensional 1170:1186 arg1 structure 1188:1196 the three-dimensional structure 1166:1196 the three-dimensional structure 1166:1196 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 2 54 theme Kanzaki 416:422 arg1 disease 424:430 Kanzaki disease 416:430 Kanzaki disease 416:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 2 54 theme Kanzaki 416:422 arg1 disorders 384:392 the lysosomal storage disorders 362:392 the lysosomal storage disorders Schindler disease and Kanzaki disease 362:430 In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. 19683538 5 55 theme active 981:986 arg1 site 988:991 the active site 977:991 the active site of the enzyme 977:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 5 55 theme active 981:986 arg1 enzyme 1000:1005 the enzyme 996:1005 the enzyme 996:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 5 56 theme human 861:865 arg1 enzyme 879:884 the human alpha-NAGAL enzyme 857:884 the human alpha-NAGAL enzyme 857:884 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 6 57 theme individual 1033:1042 arg1 defects 1044:1050 individual defects 1033:1050 individual defects in the alpha-NAGAL glycoprotein 1033:1082 To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. 19683538 5 58 theme different 939:947 arg1 products 959:966 different catalytic products 939:966 different catalytic products bound in the active site of the enzyme 939:1005 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 19683538 4 59 theme enzymatic 768:776 arg1 equivalence 778:788 their enzymatic equivalence 762:788 their enzymatic equivalence 762:788 We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. 19683538 5 60 theme catalytic 834:842 arg1 mechanism 844:852 catalytic mechanism 834:852 catalytic mechanism 834:852 To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. 16422668 6 0 dep Planque 1136:1142 arg1 Biol 1218:1221 Biol 1218:1221 Biol 1218:1221 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 10 1 theme amide 1458:1462 arg1 bonds 1464:1468 the amide bonds 1454:1468 the amide bonds of a variety of serine protease substrates 1454:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 1 2 theme antigen 215:221 arg1 Fab 201:203 the glycosylated Fab 184:203 the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia 184:308 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 1 2 theme antigen 215:221 arg1 binding 223:229 fragment antigen binding 206:229 fragment antigen binding 206:229 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 0 3 with cryoglobulin 52:63 arg1 properties 70:79 properties 70:79 properties of a natural proteolytic antibody 70:113 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 10 4 theme serine 1486:1491 arg1 substrates 1502:1511 serine protease substrates 1486:1511 serine protease substrates 1486:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 11 5 dep serine 1713:1718 arg1 triad 1755:1759 catalytic triad 1745:1759 catalytic triad 1745:1759 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 10 6 theme variety 1475:1481 arg1 bonds 1464:1468 the amide bonds 1454:1468 the amide bonds of a variety of serine protease substrates 1454:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 10 6 theme variety 1475:1481 arg1 protein 1532:1538 the gp120 coat protein 1517:1538 the gp120 coat protein of HIV 1517:1545 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 6 7 theme Yvo 1035:1037 arg1 IgM 1039:1041 Yvo IgM 1035:1041 Yvo IgM 1035:1041 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 12 8 theme bonds 1920:1924 arg1 cleavage 1900:1907 the cleavage 1896:1907 the cleavage of peptide bonds 1896:1924 Our present findings indicate that pre-existing or natural antibodies can utilize at least one novel strategy for the cleavage of peptide bonds. 16422668 0 9 theme antibody 106:113 arg1 properties 70:79 properties 70:79 properties of a natural proteolytic antibody 70:113 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 5 10 theme arms 969:972 arg1 flexibility 946:956 the segmental flexibility 932:956 the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures 932:1022 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 10 11 theme substrates 1502:1511 arg1 variety 1475:1481 a variety 1473:1481 a variety of serine protease substrates 1473:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 10 11 theme substrates 1502:1511 arg1 substrates 1502:1511 serine protease substrates 1486:1511 serine protease substrates 1486:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 11 12 theme serine 1764:1769 arg1 proteases 1771:1779 serine proteases 1764:1779 serine proteases 1764:1779 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 6 13 theme [Paul 1122:1126 arg1 properties 1065:1074 the properties 1061:1074 the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol 1061:1221 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 10 14 theme Yvo 1380:1382 arg1 protein 1384:1390 The Yvo protein 1376:1390 The Yvo protein 1376:1390 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 11 15 theme atypical 1551:1558 arg1 located 1600:1606 located 1600:1606 located 1600:1606 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 15 theme atypical 1551:1558 arg1 motif 1591:1595 An atypical serine, arginine and glutamate motif 1548:1595 motif 1591:1595 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 5 16 theme intact 977:982 arg1 IgM 988:990 intact Yvo IgM 977:990 intact Yvo IgM 977:990 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 6 17 theme proteolytic 1101:1111 arg1 Edmundson 1187:1195 Edmundson 1187:1195 Edmundson 1187:1195 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 17 theme proteolytic 1101:1111 arg1 Planque 1136:1142 Planque 1136:1142 Planque 1136:1142 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 17 theme proteolytic 1101:1111 arg1 Handy 1172:1176 Handy 1172:1176 Handy 1172:1176 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 17 theme proteolytic 1101:1111 arg1 Hanson 1201:1206 Hanson 1201:1206 Hanson (2004) 1201:1213 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 17 theme proteolytic 1101:1111 arg1 [Paul 1122:1126 a naturally occurring proteolytic antibody [Paul 1079:1126 a naturally occurring proteolytic antibody [Paul 1079:1126 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 17 theme proteolytic 1101:1111 arg1 Salas 1154:1158 Salas 1154:1158 Salas 1154:1158 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 10 18 theme coat 1527:1530 arg1 protein 1532:1538 the gp120 coat protein 1517:1538 the gp120 coat protein of HIV 1517:1545 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 4 19 theme branched 661:668 arg1 oligosaccharide 679:693 the branched N-linked oligosaccharide 657:693 the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) 657:763 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 11 20 theme classical 1703:1711 arg1 serine 1713:1718 the classical serine 1699:1718 the classical serine 1699:1718 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Bangale 1257:1263 Bangale 1257:1263 Bangale 1257:1263 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Nishiyama 1317:1325 Nishiyama 1317:1325 Nishiyama 1317:1325 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Song 1266:1269 Song 1266:1269 Song 1266:1269 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Poindexter 1288:1297 Poindexter 1288:1297 Poindexter 1288:1297 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Bick 1300:1303 Bick 1300:1303 Bick 1300:1303 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Paul 1331:1334 Paul 1331:1334 Paul 1331:1334 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Karle 1272:1276 Karle 1272:1276 Karle 1272:1276 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Taguchi 1279:1285 Taguchi 1279:1285 Taguchi 1279:1285 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 8 21 dep 39611-39619 1235:1245 arg1 Edmundson 1306:1314 Edmundson 1306:1314 Edmundson 1306:1314 279, 39611-39619; Planque, Bangale, Song, Karle, Taguchi, Poindexter, Bick, Edmundson, Nishiyama and Paul (2004) J. Biol Chem. 16422668 2 22 theme Yvo 430:432 arg1 IgM 434:436 Yvo IgM 430:436 Yvo IgM 430:436 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 11 23 theme site 1649:1652 arg1 middle 1615:1620 the middle 1611:1620 the middle of the Yvo antigen-binding site 1611:1652 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 0 24 theme IgM 48:50 arg1 cryoglobulin 52:63 an IgM cryoglobulin 45:63 an IgM cryoglobulin with properties of a natural proteolytic antibody 45:113 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 5 25 theme low 1007:1009 arg1 temperatures 1011:1022 low temperatures 1007:1022 low temperatures 1007:1022 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 4 26 dep domain 719:724 arg1 domain 742:747 first constant domain 727:747 first constant domain of heavy chain 727:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 4 26 dep domain 719:724 arg1 chain 758:762 heavy chain 752:762 heavy chain 752:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 2 27 theme gel 423:425 arg1 formation 387:395 the formation 383:395 the formation of an ordered hydroscopic gel of Yvo IgM upon cooling 383:449 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 2 28 used used 340:343 arg2 scattering 325:334 Dynamic light scattering 311:334 Dynamic light scattering 311:334 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 4 29 theme three-dimensional 626:642 arg1 structure 644:652 The three-dimensional structure 622:652 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) 622:763 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 1 30 gly glycosylated 188:199 arg1 Fab 201:203 the glycosylated Fab 184:203 the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia 184:308 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 1 30 gly glycosylated 188:199 arg1 binding 223:229 fragment antigen binding 206:229 fragment antigen binding 206:229 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 5 31 theme lateral 825:831 arg1 surface 833:839 the lateral surface 821:839 the lateral surface of the CH1 domain 821:857 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 4 32 theme CH1 715:717 arg1 domain 719:724 the CH1 domain 711:724 the CH1 domain (first constant domain of heavy chain) 711:763 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 5 33 theme CH1 848:850 arg1 domain 852:857 the CH1 domain 844:857 the CH1 domain 844:857 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 0 34 theme natural 86:92 arg1 antibody 106:113 a natural proteolytic antibody 84:113 a natural proteolytic antibody 84:113 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 5 35 theme CH1 894:896 arg1 domains 906:912 the CH1 and CH2 domains 890:912 domains 906:912 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 4 36 theme oligosaccharide 679:693 arg1 structure 644:652 The three-dimensional structure 622:652 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) 622:763 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 0 37 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 0:114 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 2 38 theme gel 361:363 arg1 point 365:369 the gel point 357:369 the gel point 357:369 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 11 39 from middle 1615:1620 arg1 located 1600:1606 located 1600:1606 located 1600:1606 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 39 from middle 1615:1620 arg1 motif 1591:1595 An atypical serine, arginine and glutamate motif 1548:1595 motif 1591:1595 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 1 40 theme IgM 238:240 arg1 Fab 201:203 the glycosylated Fab 184:203 the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia 184:308 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 1 40 theme IgM 238:240 arg1 binding 223:229 fragment antigen binding 206:229 fragment antigen binding 206:229 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 2 41 theme ordered 403:409 arg1 gel 423:425 an ordered hydroscopic gel 400:425 an ordered hydroscopic gel of Yvo IgM upon cooling 400:449 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 4 42 theme chain 758:762 arg1 domain 742:747 first constant domain 727:747 first constant domain of heavy chain 727:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 4 42 theme chain 758:762 arg1 chain 758:762 heavy chain 752:762 heavy chain 752:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 11 43 theme Yvo 1629:1631 arg1 site 1649:1652 the Yvo antigen-binding site 1625:1652 the Yvo antigen-binding site 1625:1652 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 0 44 theme glycosylated 23:34 arg1 Fab 36:38 a glycosylated Fab 21:38 a glycosylated Fab 21:38 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 12 45 theme peptide 1912:1918 arg1 bonds 1920:1924 peptide bonds 1912:1924 peptide bonds 1912:1924 Our present findings indicate that pre-existing or natural antibodies can utilize at least one novel strategy for the cleavage of peptide bonds. 16422668 5 46 from temperatures 1011:1022 arg1 flexibility 946:956 the segmental flexibility 932:956 the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures 932:1022 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 5 46 from temperatures 1011:1022 arg1 arms 969:972 the Fab arms 961:972 the Fab arms 961:972 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 5 47 theme segmental 936:944 arg1 flexibility 946:956 the segmental flexibility 932:956 the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures 932:1022 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 4 48 theme constant 733:740 arg1 domain 742:747 first constant domain 727:747 first constant domain of heavy chain 727:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 4 48 theme constant 733:740 arg1 chain 758:762 heavy chain 752:762 heavy chain 752:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 5 49 theme CH2 902:904 arg1 domains 906:912 the CH1 and CH2 domains 890:912 domains 906:912 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 3 50 theme considerable 584:595 arg1 morbidity 597:605 morbidity 597:605 morbidity 597:605 If a cryoglobulin forms gels in peripheral tissues and organs, the associated swelling and damage to microvasculature can result in considerable morbidity and mortality. 16422668 2 51 theme Dynamic 311:317 arg1 scattering 325:334 Dynamic light scattering 311:334 Dynamic light scattering 311:334 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 1 52 theme fragment 206:213 arg1 Fab 201:203 the glycosylated Fab 184:203 the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia 184:308 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 1 52 theme fragment 206:213 arg1 binding 223:229 fragment antigen binding 206:229 fragment antigen binding 206:229 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 4 53 link N-linked 670:677 arg1 oligosaccharide 679:693 the branched N-linked oligosaccharide 657:693 the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) 657:763 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 10 54 theme protease 1493:1500 arg1 substrates 1502:1511 serine protease substrates 1486:1511 serine protease substrates 1486:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 3 55 theme peripheral 484:493 arg1 tissues 495:501 peripheral tissues 484:501 peripheral tissues 484:501 If a cryoglobulin forms gels in peripheral tissues and organs, the associated swelling and damage to microvasculature can result in considerable morbidity and mortality. 16422668 10 56 theme gp120 1521:1525 arg1 protein 1532:1538 the gp120 coat protein 1517:1538 the gp120 coat protein of HIV 1517:1545 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 1 57 theme resolution 139:148 arg1 structure 150:158 The 2.6 A (1 A=0.1 nm) resolution structure 116:158 The 2.6 A (1 A=0.1 nm) resolution structure 116:158 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 12 58 theme pre-existing 1817:1828 arg1 antibodies 1841:1850 pre-existing or natural antibodies 1817:1850 pre-existing or natural antibodies 1817:1850 Our present findings indicate that pre-existing or natural antibodies can utilize at least one novel strategy for the cleavage of peptide bonds. 16422668 11 59 theme overall 1670:1676 arg1 geometry 1678:1685 an overall geometry 1667:1685 an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases 1667:1779 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 60 theme proteases 1771:1779 arg1 histidine 1721:1729 histidine 1721:1729 histidine 1721:1729 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 60 theme proteases 1771:1779 arg1 serine 1713:1718 the classical serine 1699:1718 the classical serine 1699:1718 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 60 theme proteases 1771:1779 arg1 aspartate 1735:1743 aspartate 1735:1743 aspartate 1735:1743 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 0 61 from cryoglobulin 52:63 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 0:114 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 6 62 theme antibody 1113:1120 arg1 Edmundson 1187:1195 Edmundson 1187:1195 Edmundson 1187:1195 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 62 theme antibody 1113:1120 arg1 Planque 1136:1142 Planque 1136:1142 Planque 1136:1142 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 62 theme antibody 1113:1120 arg1 Handy 1172:1176 Handy 1172:1176 Handy 1172:1176 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 62 theme antibody 1113:1120 arg1 Hanson 1201:1206 Hanson 1201:1206 Hanson (2004) 1201:1213 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 62 theme antibody 1113:1120 arg1 [Paul 1122:1126 a naturally occurring proteolytic antibody [Paul 1079:1126 a naturally occurring proteolytic antibody [Paul 1079:1126 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 62 theme antibody 1113:1120 arg1 Salas 1154:1158 Salas 1154:1158 Salas 1154:1158 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 12 63 theme natural 1833:1839 arg1 antibodies 1841:1850 pre-existing or natural antibodies 1817:1850 pre-existing or natural antibodies 1817:1850 Our present findings indicate that pre-existing or natural antibodies can utilize at least one novel strategy for the cleavage of peptide bonds. 16422668 11 64 theme serine 1560:1565 arg1 located 1600:1606 located 1600:1606 located 1600:1606 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 64 theme serine 1560:1565 arg1 motif 1591:1595 An atypical serine, arginine and glutamate motif 1548:1595 motif 1591:1595 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 5 65 theme Yvo 984:986 arg1 IgM 988:990 intact Yvo IgM 977:990 intact Yvo IgM 977:990 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 5 66 theme surface 833:839 arg1 part 813:816 part 813:816 part of the lateral surface of the CH1 domain 813:857 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 6 67 theme occurring 1091:1099 arg1 Edmundson 1187:1195 Edmundson 1187:1195 Edmundson 1187:1195 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 67 theme occurring 1091:1099 arg1 Planque 1136:1142 Planque 1136:1142 Planque 1136:1142 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 67 theme occurring 1091:1099 arg1 Handy 1172:1176 Handy 1172:1176 Handy 1172:1176 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 67 theme occurring 1091:1099 arg1 Hanson 1201:1206 Hanson 1201:1206 Hanson (2004) 1201:1213 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 67 theme occurring 1091:1099 arg1 [Paul 1122:1126 a naturally occurring proteolytic antibody [Paul 1079:1126 a naturally occurring proteolytic antibody [Paul 1079:1126 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 67 theme occurring 1091:1099 arg1 Salas 1154:1158 Salas 1154:1158 Salas 1154:1158 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 12 68 theme present 1786:1792 arg1 findings 1794:1801 Our present findings 1782:1801 Our present findings 1782:1801 Our present findings indicate that pre-existing or natural antibodies can utilize at least one novel strategy for the cleavage of peptide bonds. 16422668 12 69 theme novel 1877:1881 arg1 strategy 1883:1890 at least one novel strategy 1864:1890 at least one novel strategy 1864:1890 Our present findings indicate that pre-existing or natural antibodies can utilize at least one novel strategy for the cleavage of peptide bonds. 16422668 5 70 theme Fab 965:967 arg1 arms 969:972 the Fab arms 961:972 the Fab arms 961:972 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 3 71 theme associated 519:528 arg1 swelling 530:537 swelling 530:537 swelling 530:537 If a cryoglobulin forms gels in peripheral tissues and organs, the associated swelling and damage to microvasculature can result in considerable morbidity and mortality. 16422668 1 72 theme A 124:124 arg1 structure 150:158 The 2.6 A (1 A=0.1 nm) resolution structure 116:158 The 2.6 A (1 A=0.1 nm) resolution structure 116:158 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 5 73 from flexibility 946:956 arg1 temperatures 1011:1022 low temperatures 1007:1022 low temperatures 1007:1022 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 5 73 from flexibility 946:956 arg1 IgM 988:990 intact Yvo IgM 977:990 intact Yvo IgM 977:990 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 2 74 theme hydroscopic 411:421 arg1 gel 423:425 an ordered hydroscopic gel 400:425 an ordered hydroscopic gel of Yvo IgM upon cooling 400:449 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 11 75 located located 1600:1606 arg2 located 1600:1606 located 1600:1606 located 1600:1606 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 75 located located 1600:1606 arg1 middle 1615:1620 the middle 1611:1620 the middle of the Yvo antigen-binding site 1611:1652 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 75 located located 1600:1606 arg2 motif 1591:1595 An atypical serine, arginine and glutamate motif 1548:1595 motif 1591:1595 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 76 theme catalytic 1745:1753 arg1 triad 1755:1759 catalytic triad 1745:1759 catalytic triad 1745:1759 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 77 from located 1600:1606 arg1 middle 1615:1620 the middle 1611:1620 the middle of the Yvo antigen-binding site 1611:1652 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 10 78 theme nucleophilic 1430:1441 arg1 mechanism 1443:1451 a nucleophilic mechanism 1428:1451 a nucleophilic mechanism 1428:1451 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 0 79 theme proteolytic 94:104 arg1 antibody 106:113 a natural proteolytic antibody 84:113 a natural proteolytic antibody 84:113 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 2 80 theme IgM 434:436 arg1 gel 423:425 an ordered hydroscopic gel 400:425 an ordered hydroscopic gel of Yvo IgM upon cooling 400:449 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 5 81 from arms 969:972 arg1 IgM 988:990 intact Yvo IgM 977:990 intact Yvo IgM 977:990 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 5 82 theme domain 852:857 arg1 surface 833:839 the lateral surface 821:839 the lateral surface of the CH1 domain 821:857 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 4 83 theme N-linked 670:677 arg1 oligosaccharide 679:693 the branched N-linked oligosaccharide 657:693 the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) 657:763 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 2 84 theme light 319:323 arg1 scattering 325:334 Dynamic light scattering 311:334 Dynamic light scattering 311:334 Dynamic light scattering was used to estimate the gel point and monitor the formation of an ordered hydroscopic gel of Yvo IgM upon cooling. 16422668 5 85 from IgM 988:990 arg1 flexibility 946:956 the segmental flexibility 932:956 the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures 932:1022 The carbohydrate may act to shield part of the lateral surface of the CH1 domain and crowd the junction between the CH1 and CH2 domains, thereby limiting the segmental flexibility of the Fab arms in intact Yvo IgM, especially at low temperatures. 16422668 10 86 theme HIV 1543:1545 arg1 bonds 1464:1468 the amide bonds 1454:1468 the amide bonds of a variety of serine protease substrates 1454:1511 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 10 86 theme HIV 1543:1545 arg1 protein 1532:1538 the gp120 coat protein 1517:1538 the gp120 coat protein of HIV 1517:1545 The Yvo protein displayed the ability to cleave, by a nucleophilic mechanism, the amide bonds of a variety of serine protease substrates and the gp120 coat protein of HIV. 16422668 0 87 theme Fab 36:38 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 0:114 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 4 88 theme heavy 752:756 arg1 chain 758:762 heavy chain 752:762 heavy chain 752:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 11 89 theme antigen-binding 1633:1647 arg1 site 1649:1652 the Yvo antigen-binding site 1625:1652 the Yvo antigen-binding site 1625:1652 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 90 theme glutamate 1581:1589 arg1 located 1600:1606 located 1600:1606 located 1600:1606 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 90 theme glutamate 1581:1589 arg1 motif 1591:1595 An atypical serine, arginine and glutamate motif 1548:1595 motif 1591:1595 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 0 91 gly glycosylated 23:34 arg1 Fab 36:38 a glycosylated Fab 21:38 a glycosylated Fab 21:38 Crystal structure of a glycosylated Fab from an IgM cryoglobulin with properties of a natural proteolytic antibody. 16422668 4 92 theme first 727:731 arg1 domain 742:747 first constant domain 727:747 first constant domain of heavy chain 727:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 4 92 theme first 727:731 arg1 chain 758:762 heavy chain 752:762 heavy chain 752:762 The three-dimensional structure of the branched N-linked oligosaccharide associated with the CH1 domain (first constant domain of heavy chain) is reported. 16422668 11 93 theme arginine 1568:1575 arg1 located 1600:1606 located 1600:1606 located 1600:1606 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 11 93 theme arginine 1568:1575 arg1 motif 1591:1595 An atypical serine, arginine and glutamate motif 1548:1595 motif 1591:1595 An atypical serine, arginine and glutamate motif is located in the middle of the Yvo antigen-binding site and displays an overall geometry that mimics the classical serine, histidine and aspartate catalytic triad of serine proteases. 16422668 1 94 theme glycosylated 188:199 arg1 Fab 201:203 the glycosylated Fab 184:203 the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia 184:308 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 1 94 theme glycosylated 188:199 arg1 binding 223:229 fragment antigen binding 206:229 fragment antigen binding 206:229 The 2.6 A (1 A=0.1 nm) resolution structure has been determined for the glycosylated Fab (fragment antigen binding) of an IgM (Yvo) obtained from a subject with Waldenström's macroglobulinaemia. 16422668 6 95 contain have 1056:1059 arg1 IgM 1039:1041 Yvo IgM 1035:1041 Yvo IgM 1035:1041 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 16422668 6 95 contain have 1056:1059 arg2 properties 1065:1074 the properties 1061:1074 the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol 1061:1221 Recently, Yvo IgM was shown to have the properties of a naturally occurring proteolytic antibody [Paul, Karle, Planque, Taguchi, Salas, Nishiyama, Handy, Hunter, Edmundson and Hanson (2004) J. Biol. 11562499 0 0 theme growth-promoting 70:85 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human PEDF 0:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 0 0 theme growth-promoting 70:85 arg1 factor 87:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 4 1 theme sites 655:659 arg1 organization 601:612 the organization 597:612 the organization of possible receptor and heparin-binding sites 597:659 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 4 2 theme characterized 707:719 arg1 serpin 721:726 any other previously characterized serpin 686:726 any other previously characterized serpin 686:726 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 1 3 theme noninhibitory 139:151 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 3 theme noninhibitory 139:151 arg1 member 153:158 a noninhibitory member 137:158 a noninhibitory member of the serpin superfamily 137:184 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 0 4 theme neurite 62:68 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human PEDF 0:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 0 4 theme neurite 62:68 arg1 factor 87:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 3 5 theme many 476:479 arg1 roles 492:496 its many biological roles 472:496 its many biological roles 472:496 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 5 6 theme action 940:945 arg1 mechanisms 921:930 possible mechanisms 912:930 possible mechanisms of PEDF action 912:945 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 2 7 from activity 294:301 arg1 retina 316:321 the retina 312:321 the retina 312:321 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 2 7 from activity 294:301 arg1 system 350:355 the central nervous system 330:355 the central nervous system 330:355 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 1 8 theme mammalian 239:247 arg1 compartment 256:266 the mammalian ocular compartment 235:266 the mammalian ocular compartment 235:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 5 9 theme possible 912:919 arg1 mechanisms 921:930 possible mechanisms 912:930 possible mechanisms of PEDF action 912:945 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 1 10 theme ocular 249:254 arg1 compartment 256:266 the mammalian ocular compartment 235:266 the mammalian ocular compartment 235:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 4 11 theme functional 797:806 arg1 importance 808:817 functional importance 797:817 functional importance 797:817 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 4 12 theme possible 617:624 arg1 receptor 626:633 possible receptor 617:633 possible receptor 617:633 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 5 13 theme future 863:868 arg1 analyses 898:905 future detailed structure/function analyses 863:905 future detailed structure/function analyses into possible mechanisms of PEDF action 863:945 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 0 14 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human PEDF 0:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 0 14 theme Crystal 0:6 arg1 factor 87:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 4 15 theme other 690:694 arg1 serpin 721:726 any other previously characterized serpin 686:726 any other previously characterized serpin 686:726 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 1 16 theme serpin 167:172 arg1 superfamily 174:184 the serpin superfamily 163:184 the serpin superfamily 163:184 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 2 17 theme senescent 401:409 arg1 fibroblasts 411:421 young versus senescent fibroblasts 388:421 fibroblasts 411:421 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 3 18 theme crystal 518:524 arg1 structure 526:534 the crystal structure 514:534 the crystal structure of glycosylated human PEDF to 2.85 A 514:571 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 1 19 theme superfamily 174:184 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 19 theme superfamily 174:184 arg1 member 153:158 a noninhibitory member 137:158 a noninhibitory member of the serpin superfamily 137:184 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 0 20 theme human 21:25 arg1 PEDF 27:30 human PEDF 21:30 human PEDF 21:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 2 21 theme neurotrophic 281:292 arg1 activity 294:301 neurotrophic activity 281:301 neurotrophic activity 281:301 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 5 22 theme PEDF 935:938 arg1 action 940:945 PEDF action 935:945 PEDF action 935:945 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 2 23 theme young 388:392 arg1 fibroblasts 411:421 young versus senescent fibroblasts 388:421 fibroblasts 411:421 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 1 24 theme epithelium-derived 103:120 arg1 PEDF 130:133 PEDF 130:133 PEDF 130:133 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 24 theme epithelium-derived 103:120 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 24 theme epithelium-derived 103:120 arg1 inhibitor 206:214 the most potent inhibitor 190:214 the most potent inhibitor of angiogenesis in the mammalian ocular compartment 190:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 24 theme epithelium-derived 103:120 arg1 member 153:158 a noninhibitory member 137:158 a noninhibitory member of the serpin superfamily 137:184 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 5 25 theme starting 844:851 arg1 point 853:857 a starting point 842:857 a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis 842:1026 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 3 26 theme biological 481:490 arg1 roles 492:496 its many biological roles 472:496 its many biological roles 472:496 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 5 27 theme structure/function 879:896 arg1 analyses 898:905 future detailed structure/function analyses 863:905 future detailed structure/function analyses into possible mechanisms of PEDF action 863:945 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 3 28 theme PEDF 558:561 arg1 structure 526:534 the crystal structure 514:534 the crystal structure of glycosylated human PEDF to 2.85 A 514:571 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 2 29 theme nervous 342:348 arg1 system 350:355 the central nervous system 330:355 the central nervous system 330:355 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 0 30 theme PEDF 27:30 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human PEDF 0:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 0 30 theme PEDF 27:30 arg1 factor 87:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 2 31 contain has 277:279 arg1 It 269:270 It 269:270 It 269:270 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 2 31 contain has 277:279 arg2 activity 294:301 neurotrophic activity 281:301 neurotrophic activity 281:301 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 5 32 theme uncontrolled 1002:1013 arg1 angiogenesis 1015:1026 uncontrolled angiogenesis 1002:1026 uncontrolled angiogenesis 1002:1026 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 3 33 gly glycosylated 539:550 arg1 PEDF 558:561 glycosylated human PEDF 539:561 glycosylated human PEDF 539:561 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 0 34 theme potent 35:40 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human PEDF 0:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 0 34 theme potent 35:40 arg1 factor 87:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 4 35 contain has 734:736 arg1 PEDF 729:732 PEDF 729:732 PEDF 729:732 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 4 35 contain has 734:736 arg2 distribution 767:778 a striking asymmetric charge distribution 738:778 a striking asymmetric charge distribution that might be of functional importance 738:817 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 0 36 theme anti-angiogenic 42:56 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human PEDF 0:30 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 0 36 theme anti-angiogenic 42:56 arg1 factor 87:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 a potent anti-angiogenic and neurite growth-promoting factor 33:92 Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor. 11562499 3 37 theme glycosylated 539:550 arg1 PEDF 558:561 glycosylated human PEDF 539:561 glycosylated human PEDF 539:561 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 5 38 theme detailed 870:877 arg1 analyses 898:905 future detailed structure/function analyses 863:905 future detailed structure/function analyses into possible mechanisms of PEDF action 863:945 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 3 39 theme human 552:556 arg1 PEDF 558:561 glycosylated human PEDF 539:561 glycosylated human PEDF 539:561 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 4 40 theme receptor 626:633 arg1 organization 601:612 the organization 597:612 the organization of possible receptor and heparin-binding sites 597:659 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 5 41 theme therapeutics 981:992 arg1 development 966:976 development 966:976 development of therapeutics against uncontrolled angiogenesis 966:1026 These results provide a starting point for future detailed structure/function analyses into possible mechanisms of PEDF action that could lead to development of therapeutics against uncontrolled angiogenesis. 11562499 4 42 theme striking 740:747 arg1 distribution 767:778 a striking asymmetric charge distribution 738:778 a striking asymmetric charge distribution that might be of functional importance 738:817 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 3 43 theme structural 437:446 arg1 basis 448:452 a structural basis 435:452 a structural basis for understanding its many biological roles 435:496 To provide a structural basis for understanding its many biological roles, we have solved the crystal structure of glycosylated human PEDF to 2.85 A. 11562499 1 44 theme Pigment 95:101 arg1 PEDF 130:133 PEDF 130:133 PEDF 130:133 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 44 theme Pigment 95:101 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 44 theme Pigment 95:101 arg1 inhibitor 206:214 the most potent inhibitor 190:214 the most potent inhibitor of angiogenesis in the mammalian ocular compartment 190:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 44 theme Pigment 95:101 arg1 member 153:158 a noninhibitory member 137:158 a noninhibitory member of the serpin superfamily 137:184 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 45 theme potent 199:204 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 45 theme potent 199:204 arg1 inhibitor 206:214 the most potent inhibitor 190:214 the most potent inhibitor of angiogenesis in the mammalian ocular compartment 190:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 46 from inhibitor 206:214 arg1 compartment 256:266 the mammalian ocular compartment 235:266 the mammalian ocular compartment 235:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 4 47 theme charge 760:765 arg1 distribution 767:778 a striking asymmetric charge distribution 738:778 a striking asymmetric charge distribution that might be of functional importance 738:817 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 2 48 theme central 334:340 arg1 system 350:355 the central nervous system 330:355 the central nervous system 330:355 It also has neurotrophic activity, both in the retina and in the central nervous system, and is highly up-regulated in young versus senescent fibroblasts. 11562499 1 49 link epithelium-derived 103:120 arg1 PEDF 130:133 PEDF 130:133 PEDF 130:133 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 49 link epithelium-derived 103:120 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 49 link epithelium-derived 103:120 arg1 inhibitor 206:214 the most potent inhibitor 190:214 the most potent inhibitor of angiogenesis in the mammalian ocular compartment 190:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 49 link epithelium-derived 103:120 arg1 member 153:158 a noninhibitory member 137:158 a noninhibitory member of the serpin superfamily 137:184 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 4 50 theme asymmetric 749:758 arg1 distribution 767:778 a striking asymmetric charge distribution 738:778 a striking asymmetric charge distribution that might be of functional importance 738:817 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 4 51 theme heparin-binding 639:653 arg1 sites 655:659 heparin-binding sites 639:659 heparin-binding sites 639:659 The structure revealed the organization of possible receptor and heparin-binding sites, and showed that, unlike any other previously characterized serpin, PEDF has a striking asymmetric charge distribution that might be of functional importance. 11562499 1 52 theme angiogenesis 219:230 arg1 factor 122:127 Pigment epithelium-derived factor 95:127 Pigment epithelium-derived factor (PEDF) 95:134 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11562499 1 52 theme angiogenesis 219:230 arg1 inhibitor 206:214 the most potent inhibitor 190:214 the most potent inhibitor of angiogenesis in the mammalian ocular compartment 190:266 Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serpin superfamily, is the most potent inhibitor of angiogenesis in the mammalian ocular compartment. 11152678 9 0 from formation 1416:1424 arg1 pathway 1475:1481 the constitutive secretory pathway 1448:1481 the constitutive secretory pathway 1448:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 1 1 theme neurotrophic 186:197 arg1 pro-BDNF 217:224 pro-BDNF 217:224 pro-BDNF 217:224 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 1 1 theme neurotrophic 186:197 arg1 precursor 206:214 the brain-derived neurotrophic factor precursor 168:214 the brain-derived neurotrophic factor precursor (pro-BDNF) 168:225 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 5 2 theme BDNF 844:847 arg1 antibodies 849:858 BDNF antibodies 844:858 BDNF antibodies 844:858 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. 11152678 1 3 theme factor 199:204 arg1 pro-BDNF 217:224 pro-BDNF 217:224 pro-BDNF 217:224 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 1 3 theme factor 199:204 arg1 precursor 206:214 the brain-derived neurotrophic factor precursor 168:214 the brain-derived neurotrophic factor precursor (pro-BDNF) 168:225 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 9 4 theme constitutive 1452:1463 arg1 pathway 1475:1481 the constitutive secretory pathway 1448:1481 the constitutive secretory pathway 1448:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 9 5 theme obligatory 1385:1394 arg1 intermediate 1396:1407 an obligatory intermediate 1382:1407 an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway 1382:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 9 5 theme obligatory 1385:1394 arg1 pro-BDNF 1366:1373 28-kDa pro-BDNF 1359:1373 28-kDa pro-BDNF 1359:1373 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 8 6 theme mature 1179:1184 arg1 BDNF 1186:1189 mature BDNF 1179:1189 mature BDNF 1179:1189 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 6 7 theme Arg-Gly-Leu-Thr 979:993 arg1 site 1023:1026 the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 975:1026 the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 975:1026 This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site. 11152678 2 8 theme SDS-polyacrylamide 331:348 arg1 electrophoresis 354:368 SDS-polyacrylamide gel electrophoresis 331:368 SDS-polyacrylamide gel electrophoresis 331:368 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 5 9 theme 28-kDa 797:802 arg1 protein 804:810 a 28-kDa protein 795:810 a 28-kDa protein that is immunoprecipitated with BDNF antibodies 795:858 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. 11152678 2 10 theme 32-kDa 409:414 arg1 pro-BDNF 382:389 pro-BDNF 382:389 pro-BDNF 382:389 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 10 theme 32-kDa 409:414 arg1 precursor 416:424 a 32-kDa precursor 407:424 a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain 407:498 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 10 theme 32-kDa 409:414 arg1 N-glycosylated 434:447 N-glycosylated 434:447 N-glycosylated 434:447 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 4 11 theme N-terminal 658:667 arg1 cleavage 669:676 N-terminal cleavage 658:676 N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa) 658:775 The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). 11152678 5 12 theme protein 804:810 arg1 protein 804:810 a 28-kDa protein 795:810 a 28-kDa protein that is immunoprecipitated with BDNF antibodies 795:858 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. 11152678 5 12 theme protein 804:810 arg1 amounts 784:790 Small amounts 778:790 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies 778:858 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. 11152678 8 13 contain has 1149:1151 arg1 generation 1123:1132 Blocking generation 1114:1132 Blocking generation of 28-kDa BDNF 1114:1147 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 8 13 contain has 1149:1151 arg2 effect 1156:1161 no effect 1153:1161 no effect 1153:1161 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 1 14 theme precursor 206:214 arg1 processing 154:163 post-translational processing 135:163 post-translational processing 135:163 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 1 14 theme precursor 206:214 arg1 biosynthesis 118:129 biosynthesis 118:129 biosynthesis 118:129 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 8 15 theme BDNF 1186:1189 arg1 level 1170:1174 the level 1166:1174 the level of mature BDNF 1166:1189 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 6 16 theme pro-BDNF 963:970 arg1 cleavage 951:958 N-terminal cleavage 940:958 N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 940:1026 This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site. 11152678 1 17 from processing 154:163 arg1 cells 230:234 cells 230:234 cells infected with a pro-BDNF-encoding vaccinia virus 230:283 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 8 18 theme form 1329:1332 arg1 accumulation 1302:1313 accumulation 1302:1313 accumulation of the 28-kDa form 1302:1332 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 8 19 theme blocking 1195:1202 arg1 generation 1204:1213 blocking generation 1195:1213 blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, 1195:1283 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 2 20 gly N-glycosylated 434:447 arg2 site 472:475 a site 470:475 a site 470:475 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg2 pro-BDNF 382:389 pro-BDNF 382:389 pro-BDNF 382:389 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg1 precursor 416:424 a 32-kDa precursor 407:424 a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain 407:498 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg2 N-glycosylated 434:447 N-glycosylated 434:447 N-glycosylated 434:447 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg1 pro-BDNF 382:389 pro-BDNF 382:389 pro-BDNF 382:389 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg1 site 472:475 a site 470:475 a site 470:475 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg2 precursor 416:424 a 32-kDa precursor 407:424 a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain 407:498 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 2 20 gly N-glycosylated 434:447 arg1 N-glycosylated 434:447 N-glycosylated 434:447 N-glycosylated 434:447 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 4 21 theme mature 756:761 arg1 kDa 772:774 14 kDa 769:774 14 kDa 769:774 The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). 11152678 4 21 theme mature 756:761 arg1 BDNF 763:766 mature BDNF 756:766 mature BDNF (14 kDa) 756:775 The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). 11152678 8 22 theme mature 1218:1223 arg1 BDNF 1225:1228 mature BDNF 1218:1228 mature BDNF 1218:1228 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 8 23 theme Blocking 1114:1121 arg1 generation 1123:1132 Blocking generation 1114:1132 Blocking generation of 28-kDa BDNF 1114:1147 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 3 24 theme TrkB 612:615 arg1 phosphorylation 617:631 TrkB phosphorylation 612:631 TrkB phosphorylation 612:631 Some pro-BDNF is released extracellularly and is biologically active as demonstrated by its ability to mediate TrkB phosphorylation. 11152678 4 25 theme immature 716:723 arg1 vesicles 735:742 immature secretory vesicles 716:742 immature secretory vesicles 716:742 The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). 11152678 0 26 theme post-translational 17:34 arg1 processing 36:45 post-translational processing 17:45 post-translational processing 17:45 Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. 11152678 5 27 theme Small 778:782 arg1 protein 804:810 a 28-kDa protein 795:810 a 28-kDa protein that is immunoprecipitated with BDNF antibodies 795:858 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. 11152678 5 27 theme Small 778:782 arg1 amounts 784:790 Small amounts 778:790 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies 778:858 Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. 11152678 2 28 theme gel 350:352 arg1 electrophoresis 354:368 SDS-polyacrylamide gel electrophoresis 331:368 SDS-polyacrylamide gel electrophoresis 331:368 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 9 29 theme form 1440:1443 arg1 formation 1416:1424 the formation 1412:1424 the formation of the 14-kDa form in the constitutive secretory pathway 1412:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 8 30 theme enzymes 1276:1282 arg1 inhibitor 1252:1260 an inhibitor 1249:1260 an inhibitor of furin-like enzymes 1249:1282 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 8 30 theme enzymes 1276:1282 arg1 alpha 1235:1239 alpha(1)-PDX 1235:1246 alpha(1)-PDX 1235:1246 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 6 31 theme N-terminal 940:949 arg1 cleavage 951:958 N-terminal cleavage 940:958 N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 940:1026 This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site. 11152678 8 32 theme furin-like 1265:1274 arg1 enzymes 1276:1282 furin-like enzymes 1265:1282 furin-like enzymes 1265:1282 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 8 33 theme BDNF 1225:1228 arg1 generation 1204:1213 blocking generation 1195:1213 blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, 1195:1283 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 1 34 link brain-derived 172:184 arg1 pro-BDNF 217:224 pro-BDNF 217:224 pro-BDNF 217:224 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 1 34 link brain-derived 172:184 arg1 precursor 206:214 the brain-derived neurotrophic factor precursor 168:214 the brain-derived neurotrophic factor precursor (pro-BDNF) 168:225 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 4 35 theme secretory 725:733 arg1 vesicles 735:742 immature secretory vesicles 716:742 immature secretory vesicles 716:742 The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). 11152678 6 36 theme arrow-Ser-Leu 1009:1021 arg1 site 1023:1026 the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 975:1026 the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 975:1026 This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site. 11152678 8 37 theme BDNF 1144:1147 arg1 generation 1123:1132 Blocking generation 1114:1132 Blocking generation of 28-kDa BDNF 1114:1147 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 7 38 dep in 1092:1093 arg1 vitro 1095:1099 vitro 1095:1099 vitro 1095:1099 Cleavage is abolished when Arg(54) is changed to Ala (R54A) by in vitro mutagenesis. 11152678 7 39 theme in 1092:1093 arg1 mutagenesis 1101:1111 in vitro mutagenesis 1092:1111 in vitro mutagenesis 1092:1111 Cleavage is abolished when Arg(54) is changed to Ala (R54A) by in vitro mutagenesis. 11152678 0 40 theme precursor 54:62 arg1 Biosynthesis 0:11 Biosynthesis 0:11 Biosynthesis 0:11 Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. 11152678 0 40 theme precursor 54:62 arg1 processing 36:45 post-translational processing 17:45 post-translational processing 17:45 Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. 11152678 8 41 theme 28-kDa 1137:1142 arg1 BDNF 1144:1147 28-kDa BDNF 1137:1147 28-kDa BDNF 1137:1147 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 6 42 theme endoplasmic 910:920 arg1 reticulum 922:930 the endoplasmic reticulum 906:930 the endoplasmic reticulum 906:930 This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site. 11152678 8 43 with generation 1204:1213 arg1 inhibitor 1252:1260 an inhibitor 1249:1260 an inhibitor of furin-like enzymes 1249:1282 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 8 43 with generation 1204:1213 arg1 alpha 1235:1239 alpha(1)-PDX 1235:1246 alpha(1)-PDX 1235:1246 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 1 44 theme post-translational 135:152 arg1 processing 154:163 post-translational processing 135:163 post-translational processing 135:163 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 6 45 from site 1023:1026 arg1 cleavage 951:958 N-terminal cleavage 940:958 N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site 940:1026 This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr(57)- downward arrow-Ser-Leu site. 11152678 9 46 from intermediate 1396:1407 arg1 formation 1416:1424 the formation 1412:1424 the formation of the 14-kDa form in the constitutive secretory pathway 1412:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 0 47 link brain-derived 67:79 arg1 factor 94:99 brain-derived neurotrophic factor 67:99 brain-derived neurotrophic factor 67:99 Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. 11152678 1 48 from biosynthesis 118:129 arg1 cells 230:234 cells 230:234 cells infected with a pro-BDNF-encoding vaccinia virus 230:283 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 9 49 theme 28-kDa 1359:1364 arg1 intermediate 1396:1407 an obligatory intermediate 1382:1407 an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway 1382:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 9 49 theme 28-kDa 1359:1364 arg1 pro-BDNF 1366:1373 28-kDa pro-BDNF 1359:1373 28-kDa pro-BDNF 1359:1373 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 8 50 theme 28-kDa 1322:1327 arg1 form 1329:1332 the 28-kDa form 1318:1332 the 28-kDa form 1318:1332 Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with alpha(1)-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. 11152678 1 51 theme brain-derived 172:184 arg1 pro-BDNF 217:224 pro-BDNF 217:224 pro-BDNF 217:224 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 1 51 theme brain-derived 172:184 arg1 precursor 206:214 the brain-derived neurotrophic factor precursor 168:214 the brain-derived neurotrophic factor precursor (pro-BDNF) 168:225 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 1 52 theme pro-BDNF-encoding 252:268 arg1 virus 279:283 a pro-BDNF-encoding vaccinia virus 250:283 a pro-BDNF-encoding vaccinia virus 250:283 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 0 53 theme neurotrophic 81:92 arg1 factor 94:99 brain-derived neurotrophic factor 67:99 brain-derived neurotrophic factor 67:99 Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. 11152678 2 54 theme Metabolic 286:294 arg1 labeling 296:303 Metabolic labeling 286:303 Metabolic labeling 286:303 Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. 11152678 9 55 theme 14-kDa 1433:1438 arg1 form 1440:1443 the 14-kDa form 1429:1443 the 14-kDa form 1429:1443 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 9 56 theme secretory 1465:1473 arg1 pathway 1475:1481 the constitutive secretory pathway 1448:1481 the constitutive secretory pathway 1448:1481 These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. 11152678 1 57 theme vaccinia 270:277 arg1 virus 279:283 a pro-BDNF-encoding vaccinia virus 250:283 a pro-BDNF-encoding vaccinia virus 250:283 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 0 58 theme brain-derived 67:79 arg1 factor 94:99 brain-derived neurotrophic factor 67:99 brain-derived neurotrophic factor 67:99 Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. 11152678 1 59 dep biosynthesis 118:129 arg1 the 114:116 the 114:116 the 114:116 We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. 11152678 4 60 theme trans-Golgi 689:699 arg1 network 701:707 the trans-Golgi network 685:707 the trans-Golgi network 685:707 The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). 1993171 11 0 theme Complete 1349:1356 arg1 alkylation 1358:1367 Complete alkylation 1349:1367 Complete alkylation of mouse IL-4 under mild conditions 1349:1403 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 10 1 theme mammary 1328:1334 arg1 cells 1342:1346 C127 mammary tumor cells 1323:1346 C127 mammary tumor cells 1323:1346 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 1 2 theme proteins 131:138 arg1 pairings 82:89 The disulfide pairings 68:89 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins 68:138 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins have been determined. 1993171 10 3 used used 1315:1318 arg2 sites 1306:1310 the potential N-glycosylation sites 1276:1310 the potential N-glycosylation sites 1276:1310 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 3 used used 1315:1318 arg2 one 1269:1271 one 1269:1271 one 1269:1271 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 4 theme third 1201:1205 arg1 cysteines 1217:1225 the third and fifth cysteines 1197:1225 the third and fifth cysteines of human IL-4 1197:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 5 theme sites 1306:1310 arg1 one 1269:1271 one 1269:1271 one 1269:1271 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 5 theme sites 1306:1310 arg1 sites 1306:1310 the potential N-glycosylation sites 1276:1310 the potential N-glycosylation sites 1276:1310 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 0 6 from assignments 10:20 arg1 human 47:51 human 47:51 human 47:51 Disulfide assignments in recombinant mouse and human interleukin 4. 1993171 0 6 from assignments 10:20 arg1 mouse 37:41 recombinant mouse 25:41 recombinant mouse 25:41 Disulfide assignments in recombinant mouse and human interleukin 4. 1993171 8 7 theme first 980:984 arg1 cysteines 1036:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 3 8 theme linear 487:492 arg1 gradients 511:519 linear acetonitrile-TFA gradients 487:519 linear acetonitrile-TFA gradients 487:519 Peptide maps were produced by digesting the proteins with pepsin and separating the resulting fragments by reverse-phase HPLC using linear acetonitrile-TFA gradients. 1993171 10 9 theme IL-4 1236:1239 arg1 cysteines 1217:1225 the third and fifth cysteines 1197:1225 the third and fifth cysteines of human IL-4 1197:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 4 10 theme reverse-phase 587:599 arg1 peaks 601:605 peaks 601:605 peaks 601:605 Cystine-containing peptides were identified by determining which reverse-phase peaks showed an altered elution pattern after reduction. 1993171 11 11 theme mild 1389:1392 arg1 conditions 1394:1403 mild conditions 1389:1403 mild conditions 1389:1403 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 11 12 theme hematopoietic 1455:1467 arg1 assay 1498:1502 a hematopoietic precursor cell proliferation assay 1453:1502 a hematopoietic precursor cell proliferation assay 1453:1502 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 7 13 theme third 892:896 arg1 cysteines 908:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 For mouse IL-4, the first and fifth, second and fourth, and third and sixth cysteines are joined. 1993171 10 14 theme N-glycosylation 1290:1304 arg1 sites 1306:1310 the potential N-glycosylation sites 1276:1310 the potential N-glycosylation sites 1276:1310 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 8 15 theme third 1020:1024 arg1 cysteines 1036:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 8 16 theme sixth 990:994 arg1 cysteines 1036:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 10 17 theme potential 1280:1288 arg1 sites 1306:1310 the potential N-glycosylation sites 1276:1310 the potential N-glycosylation sites 1276:1310 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 9 18 theme large 1049:1053 arg1 region 1067:1072 A large double-loop region 1047:1072 A large double-loop region within the central three-fifths of each protein 1047:1120 A large double-loop region within the central three-fifths of each protein is stabilized by these bonds. 1993171 10 19 theme peptides 1177:1184 arg1 analysis 1161:1168 Sequence analysis 1152:1168 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 1152:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 11 20 theme IL-4 1378:1381 arg1 alkylation 1358:1367 Complete alkylation 1349:1367 Complete alkylation of mouse IL-4 under mild conditions 1349:1403 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 3 21 theme reverse-phase 462:474 arg1 HPLC 476:479 reverse-phase HPLC 462:479 reverse-phase HPLC using linear acetonitrile-TFA gradients 462:519 Peptide maps were produced by digesting the proteins with pepsin and separating the resulting fragments by reverse-phase HPLC using linear acetonitrile-TFA gradients. 1993171 3 22 theme resulting 439:447 arg1 fragments 449:457 the resulting fragments 435:457 the resulting fragments 435:457 Peptide maps were produced by digesting the proteins with pepsin and separating the resulting fragments by reverse-phase HPLC using linear acetonitrile-TFA gradients. 1993171 9 23 theme double-loop 1055:1065 arg1 region 1067:1072 A large double-loop region 1047:1072 A large double-loop region within the central three-fifths of each protein 1047:1120 A large double-loop region within the central three-fifths of each protein is stabilized by these bonds. 1993171 8 24 theme second 997:1002 arg1 cysteines 1036:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 the first and sixth, second and fourth, and third and fifth cysteines 976:1044 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 1 25 theme disulfide 72:80 arg1 pairings 82:89 The disulfide pairings 68:89 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins 68:138 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins have been determined. 1993171 7 26 theme second 869:874 arg1 cysteines 908:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 For mouse IL-4, the first and fifth, second and fourth, and third and sixth cysteines are joined. 1993171 8 27 from bonds 944:948 arg1 IL-4 959:962 human IL-4 953:962 human IL-4 953:962 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 3 28 theme Peptide 355:361 arg1 maps 363:366 Peptide maps 355:366 Peptide maps 355:366 Peptide maps were produced by digesting the proteins with pepsin and separating the resulting fragments by reverse-phase HPLC using linear acetonitrile-TFA gradients. 1993171 10 29 contain containing 1186:1195 arg1 peptides 1177:1184 the peptides 1173:1184 the peptides containing the third and fifth cysteines of human IL-4 1173:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 29 contain containing 1186:1195 arg2 cysteines 1217:1225 the third and fifth cysteines 1197:1225 the third and fifth cysteines of human IL-4 1197:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 0 30 theme Disulfide 0:8 arg1 assignments 10:20 Disulfide assignments 0:20 Disulfide assignments in recombinant mouse and human interleukin 4. 0:66 Disulfide assignments in recombinant mouse and human interleukin 4. 1993171 7 31 theme fifth 862:866 arg1 cysteines 908:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 For mouse IL-4, the first and fifth, second and fourth, and third and sixth cysteines are joined. 1993171 2 32 theme assays 347:352 arg1 variety 325:331 a variety 323:331 a variety of functional assays 323:352 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 2 32 theme assays 347:352 arg1 assays 347:352 functional assays 336:352 functional assays 336:352 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 0 33 theme recombinant 25:35 arg1 mouse 37:41 recombinant mouse 25:41 recombinant mouse 25:41 Disulfide assignments in recombinant mouse and human interleukin 4. 1993171 5 34 theme sequence 726:733 arg1 analysis 735:742 sequence analysis 726:742 sequence analysis 726:742 These peptides were purified further and defined by composition and sequence analysis. 1993171 1 35 theme mouse 94:98 arg1 pairings 82:89 The disulfide pairings 68:89 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins 68:138 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins have been determined. 1993171 9 36 theme central 1085:1091 arg1 three-fifths 1093:1104 the central three-fifths 1081:1104 the central three-fifths of each protein 1081:1120 A large double-loop region within the central three-fifths of each protein is stabilized by these bonds. 1993171 8 37 theme human 953:957 arg1 IL-4 959:962 human IL-4 953:962 human IL-4 953:962 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 7 38 theme mouse 836:840 arg1 IL-4 842:845 mouse IL-4 836:845 mouse IL-4 836:845 For mouse IL-4, the first and fifth, second and fourth, and third and sixth cysteines are joined. 1993171 7 39 theme first 852:856 arg1 cysteines 908:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 the first and fifth, second and fourth, and third and sixth cysteines 848:916 For mouse IL-4, the first and fifth, second and fourth, and third and sixth cysteines are joined. 1993171 3 40 theme acetonitrile-TFA 494:509 arg1 gradients 511:519 linear acetonitrile-TFA gradients 487:519 linear acetonitrile-TFA gradients 487:519 Peptide maps were produced by digesting the proteins with pepsin and separating the resulting fragments by reverse-phase HPLC using linear acetonitrile-TFA gradients. 1993171 11 41 theme biological 1430:1439 arg1 activity 1441:1448 its biological activity 1426:1448 its biological activity 1426:1448 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 4 42 theme Cystine-containing 522:539 arg1 peptides 541:548 Cystine-containing peptides 522:548 Cystine-containing peptides 522:548 Cystine-containing peptides were identified by determining which reverse-phase peaks showed an altered elution pattern after reduction. 1993171 11 43 theme precursor 1469:1477 arg1 assay 1498:1502 a hematopoietic precursor cell proliferation assay 1453:1502 a hematopoietic precursor cell proliferation assay 1453:1502 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 2 44 theme DNA 212:214 arg1 technology 216:225 recombinant DNA technology 200:225 recombinant DNA technology 200:225 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 8 45 theme disulfide 934:942 arg1 bonds 944:948 The disulfide bonds 930:948 The disulfide bonds in human IL-4 930:962 The disulfide bonds in human IL-4 are between the first and sixth, second and fourth, and third and fifth cysteines. 1993171 10 46 gly N-glycosylation 1290:1304 arg2 sites 1306:1310 the potential N-glycosylation sites 1276:1310 the potential N-glycosylation sites 1276:1310 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 2 47 theme recombinant 200:210 arg1 technology 216:225 recombinant DNA technology 200:225 recombinant DNA technology 200:225 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 9 48 theme protein 1114:1120 arg1 three-fifths 1093:1104 the central three-fifths 1081:1104 the central three-fifths of each protein 1081:1120 A large double-loop region within the central three-fifths of each protein is stabilized by these bonds. 1993171 1 49 theme human 104:108 arg1 proteins 131:138 human interleukin 4 (IL-4) proteins 104:138 human interleukin 4 (IL-4) proteins 104:138 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins have been determined. 1993171 6 50 link disulfide-linked 759:774 arg1 peptides 776:783 disulfide-linked peptides 759:783 disulfide-linked peptides 759:783 Three sets of disulfide-linked peptides were consistently identified for each protein. 1993171 3 51 with proteins 399:406 arg1 pepsin 413:418 pepsin 413:418 pepsin 413:418 Peptide maps were produced by digesting the proteins with pepsin and separating the resulting fragments by reverse-phase HPLC using linear acetonitrile-TFA gradients. 1993171 0 52 dep mouse 37:41 arg1 interleukin 53:63 interleukin 4 53:65 interleukin 4 53:65 Disulfide assignments in recombinant mouse and human interleukin 4. 1993171 1 53 theme interleukin 110:120 arg1 proteins 131:138 human interleukin 4 (IL-4) proteins 104:138 human interleukin 4 (IL-4) proteins 104:138 The disulfide pairings of mouse and human interleukin 4 (IL-4) proteins have been determined. 1993171 11 54 theme proliferation 1484:1496 arg1 assay 1498:1502 a hematopoietic precursor cell proliferation assay 1453:1502 a hematopoietic precursor cell proliferation assay 1453:1502 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 2 55 theme biological 300:309 arg1 response 311:318 an appropriate biological response 285:318 an appropriate biological response 285:318 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 4 56 theme elution 625:631 arg1 pattern 633:639 an altered elution pattern 614:639 an altered elution pattern 614:639 Cystine-containing peptides were identified by determining which reverse-phase peaks showed an altered elution pattern after reduction. 1993171 10 57 theme human 1230:1234 arg1 IL-4 1236:1239 human IL-4 1230:1239 human IL-4 1230:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 58 theme C127 1323:1326 arg1 cells 1342:1346 C127 mammary tumor cells 1323:1346 C127 mammary tumor cells 1323:1346 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 6 59 theme peptides 776:783 arg1 sets 751:754 Three sets 745:754 Three sets of disulfide-linked peptides 745:783 Three sets of disulfide-linked peptides were consistently identified for each protein. 1993171 6 59 theme peptides 776:783 arg1 peptides 776:783 disulfide-linked peptides 759:783 disulfide-linked peptides 759:783 Three sets of disulfide-linked peptides were consistently identified for each protein. 1993171 2 60 theme appropriate 288:298 arg1 response 311:318 an appropriate biological response 285:318 an appropriate biological response 285:318 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 10 61 theme fifth 1211:1215 arg1 cysteines 1217:1225 the third and fifth cysteines 1197:1225 the third and fifth cysteines of human IL-4 1197:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 4 62 theme altered 617:623 arg1 pattern 633:639 an altered elution pattern 614:639 an altered elution pattern 614:639 Cystine-containing peptides were identified by determining which reverse-phase peaks showed an altered elution pattern after reduction. 1993171 2 63 theme purified 166:173 arg1 proteins 175:182 The purified proteins 162:182 The purified proteins 162:182 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 2 63 theme purified 166:173 arg1 active 238:243 active 238:243 active 238:243 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 6 64 theme disulfide-linked 759:774 arg1 peptides 776:783 disulfide-linked peptides 759:783 disulfide-linked peptides 759:783 Three sets of disulfide-linked peptides were consistently identified for each protein. 1993171 2 65 theme functional 336:345 arg1 assays 347:352 functional assays 336:352 functional assays 336:352 The purified proteins, synthesized by recombinant DNA technology, are fully active as judged by their ability to stimulate an appropriate biological response in a variety of functional assays. 1993171 11 66 theme cell 1479:1482 arg1 assay 1498:1502 a hematopoietic precursor cell proliferation assay 1453:1502 a hematopoietic precursor cell proliferation assay 1453:1502 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 11 67 theme mouse 1372:1376 arg1 IL-4 1378:1381 mouse IL-4 1372:1381 mouse IL-4 1372:1381 Complete alkylation of mouse IL-4 under mild conditions completely destroyed its biological activity in a hematopoietic precursor cell proliferation assay. 1993171 10 68 theme tumor 1336:1340 arg1 cells 1342:1346 C127 mammary tumor cells 1323:1346 C127 mammary tumor cells 1323:1346 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 1993171 10 69 theme Sequence 1152:1159 arg1 analysis 1161:1168 Sequence analysis 1152:1168 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 1152:1239 Sequence analysis of the peptides containing the third and fifth cysteines of human IL-4 also demonstrated that only one of the potential N-glycosylation sites is used by C127 mammary tumor cells. 12600216 6 0 theme spanning 873:880 arg1 helices 882:888 the various membrane spanning helices 852:888 the various membrane spanning helices 852:888 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 8 1 dep sets 1248:1251 arg1 each 1260:1263 each 1260:1263 each 1260:1263 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 6 2 theme various 856:862 arg1 helices 882:888 the various membrane spanning helices 852:888 the various membrane spanning helices 852:888 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 8 3 theme NCKX 1217:1220 arg1 topology 1222:1229 a consistent NCKX topology 1204:1229 a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices 1204:1302 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 10 4 theme NCX 1519:1521 arg1 model 1532:1536 the current NCX topology model 1507:1536 the current NCX topology model with respect to the C-terminal three membrane helices 1507:1590 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 1 5 theme critical 128:135 arg1 role 137:140 a critical role 126:140 a critical role 126:140 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 7 6 theme connecting 1003:1012 arg1 loops 1014:1018 the connecting loops 999:1018 the connecting loops 999:1018 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 7 gly N-glycosylation 1153:1167 arg2 sites 1169:1173 N-glycosylation sites 1153:1173 N-glycosylation sites 1153:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 8 8 theme TMs 1256:1258 arg1 TMs 1256:1258 TMs 1256:1258 TMs 1256:1258 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 8 8 theme TMs 1256:1258 arg1 sets 1248:1251 the two sets 1240:1251 the two sets of TMs each 1240:1263 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 10 9 theme C-terminal 1558:1567 arg1 helices 1584:1590 the C-terminal three membrane helices 1554:1590 the C-terminal three membrane helices 1554:1590 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 4 10 from N-terminus 558:567 arg1 located 527:533 located 527:533 located 527:533 The first large hydrophilic loop is located extracellularly at the N-terminus; the other is cytoplasmic and separates the two sets of TMs. 12600216 10 11 from model 1532:1536 arg1 different 1492:1500 different 1492:1500 different 1492:1500 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 7 12 theme cysteine 1118:1125 arg1 residues 1127:1134 substituted cysteine residues 1106:1134 substituted cysteine residues 1106:1134 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 9 13 theme spanning 1355:1362 arg1 helix 1364:1368 membrane spanning helix six 1346:1372 membrane spanning helix six 1346:1372 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 9 14 from helix 1364:1368 arg1 cytoplasm 1381:1389 the cytoplasm 1377:1389 the cytoplasm 1377:1389 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 4 15 theme TMs 625:627 arg1 sets 617:620 the two sets 609:620 the two sets of TMs 609:627 The first large hydrophilic loop is located extracellularly at the N-terminus; the other is cytoplasmic and separates the two sets of TMs. 12600216 4 15 theme TMs 625:627 arg1 TMs 625:627 TMs 625:627 TMs 625:627 The first large hydrophilic loop is located extracellularly at the N-terminus; the other is cytoplasmic and separates the two sets of TMs. 12600216 3 16 theme exchanger 373:381 arg1 topology 349:356 The topology 345:356 The topology of the Na/Ca-K exchanger 345:381 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 2 17 located found 267:271 arg1 cones 276:280 cones 276:280 cones 276:280 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 2 17 located found 267:271 arg2 isoform 256:262 the NCKX2 isoform 246:262 the NCKX2 isoform 246:262 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 4 18 theme large 501:505 arg1 loop 519:522 The first large hydrophilic loop 491:522 The first large hydrophilic loop 491:522 The first large hydrophilic loop is located extracellularly at the N-terminus; the other is cytoplasmic and separates the two sets of TMs. 12600216 1 19 theme Na/Ca-K 58:64 arg1 NCKX 77:80 NCKX 77:80 NCKX 77:80 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 1 19 theme Na/Ca-K 58:64 arg1 exchanger 66:74 The Na/Ca-K exchanger 54:74 The Na/Ca-K exchanger (NCKX) 54:81 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 1 19 theme Na/Ca-K 58:64 arg1 protein 107:113 a polytopic membrane protein 86:113 a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors 86:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 2 20 theme ganglion 294:301 arg1 cells 303:307 retinal ganglion cells 286:307 retinal ganglion cells 286:307 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 9 21 theme new 1309:1311 arg1 model 1313:1317 Our new model 1305:1317 Our new model 1305:1317 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 10 22 theme topology 1474:1481 arg1 model 1483:1487 this NCKX topology model 1464:1487 this NCKX topology model 1464:1487 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 3 23 theme hydrophilic 418:428 arg1 loops 430:434 two large hydrophilic loops 408:434 two large hydrophilic loops 408:434 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 5 24 theme membrane 712:719 arg1 helices 730:736 five and six membrane spanning helices 699:736 five and six membrane spanning helices 699:736 The TMs consist of either five and five membrane spanning helices or five and six membrane spanning helices, depending upon the predictive algorithm used. 12600216 7 25 theme residues 1127:1134 arg1 methods 1080:1086 two different methods 1066:1086 two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites 1066:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 25 theme residues 1127:1134 arg1 accessibility 1089:1101 accessibility 1089:1101 accessibility of substituted cysteine residues 1089:1134 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 25 theme residues 1127:1134 arg1 insertion 1140:1148 insertion 1140:1148 insertion of N-glycosylation sites 1140:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 1 26 theme Ca 145:146 arg1 homeostasis 152:162 Ca(2+) homeostasis 145:162 Ca(2+) homeostasis in retinal rod and cone photoreceptors 145:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 2 27 theme brain 338:342 arg1 parts 325:329 various parts 317:329 various parts of the brain 317:342 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 9 28 theme membrane 1346:1353 arg1 helix 1364:1368 membrane spanning helix six 1346:1372 membrane spanning helix six 1346:1372 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 3 29 theme transmembrane 452:464 arg1 TMs 485:487 TMs 485:487 TMs 485:487 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 3 29 theme transmembrane 452:464 arg1 segments 475:482 transmembrane spanning segments 452:482 transmembrane spanning segments (TMs) 452:488 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 7 30 dep methods 1080:1086 arg1 methods 1080:1086 two different methods 1066:1086 two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites 1066:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 30 dep methods 1080:1086 arg1 accessibility 1089:1101 accessibility 1089:1101 accessibility of substituted cysteine residues 1089:1134 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 30 dep methods 1080:1086 arg1 insertion 1140:1148 insertion 1140:1148 insertion of N-glycosylation sites 1140:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 2 31 theme retinal 286:292 arg1 cells 303:307 retinal ganglion cells 286:307 retinal ganglion cells 286:307 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 0 32 theme retinal 16:22 arg1 exchanger 43:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 Topology of the retinal cone NCKX2 Na/Ca-K exchanger. 12600216 10 33 theme topology 1523:1530 arg1 model 1532:1536 the current NCX topology model 1507:1536 the current NCX topology model with respect to the C-terminal three membrane helices 1507:1590 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 9 34 dep helix 1364:1368 arg1 places 1398:1403 places 1398:1403 places the C-terminus on the extracellular surface 1398:1447 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 4 35 theme hydrophilic 507:517 arg1 loop 519:522 The first large hydrophilic loop 491:522 The first large hydrophilic loop 491:522 The first large hydrophilic loop is located extracellularly at the N-terminus; the other is cytoplasmic and separates the two sets of TMs. 12600216 5 36 theme spanning 679:686 arg1 helices 688:694 five and five membrane spanning helices 656:694 five and five membrane spanning helices 656:694 The TMs consist of either five and five membrane spanning helices or five and six membrane spanning helices, depending upon the predictive algorithm used. 12600216 3 37 theme large 412:416 arg1 loops 430:434 two large hydrophilic loops 408:434 two large hydrophilic loops 408:434 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 2 38 theme various 317:323 arg1 parts 325:329 various parts 317:329 various parts of the brain 317:342 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 0 39 theme NCKX2 29:33 arg1 exchanger 43:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 Topology of the retinal cone NCKX2 Na/Ca-K exchanger. 12600216 8 40 contain contain 1265:1271 arg1 TMs 1256:1258 TMs 1256:1258 TMs 1256:1258 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 8 40 contain contain 1265:1271 arg1 topology 1222:1229 a consistent NCKX topology 1204:1229 a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices 1204:1302 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 8 40 contain contain 1265:1271 arg2 sets 1248:1251 the two sets 1240:1251 the two sets of TMs each 1240:1263 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 8 40 contain contain 1265:1271 arg2 TMs 1256:1258 TMs 1256:1258 TMs 1256:1258 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 8 40 contain contain 1265:1271 arg2 helices 1296:1302 five membrane spanning helices 1273:1302 five membrane spanning helices 1273:1302 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 8 40 contain contain 1265:1271 arg1 sets 1248:1251 the two sets 1240:1251 the two sets of TMs each 1240:1263 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 9 41 theme extracellular 1427:1439 arg1 surface 1441:1447 the extracellular surface 1423:1447 the extracellular surface 1423:1447 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 6 42 theme short 918:922 arg1 loops 924:928 the short loops 914:928 the short loops connecting these helices 914:953 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 10 43 theme current 1511:1517 arg1 model 1532:1536 the current NCX topology model 1507:1536 the current NCX topology model with respect to the C-terminal three membrane helices 1507:1590 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 0 44 theme cone 24:27 arg1 exchanger 43:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 Topology of the retinal cone NCKX2 Na/Ca-K exchanger. 12600216 2 45 located found 225:229 arg2 isoform 214:220 The NCKX1 isoform 204:220 The NCKX1 isoform 204:220 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 2 45 located found 225:229 arg1 rods 234:237 rods 234:237 rods 234:237 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 5 46 theme spanning 721:728 arg1 helices 730:736 five and six membrane spanning helices 699:736 five and six membrane spanning helices 699:736 The TMs consist of either five and five membrane spanning helices or five and six membrane spanning helices, depending upon the predictive algorithm used. 12600216 5 47 theme membrane 670:677 arg1 helices 688:694 five and five membrane spanning helices 656:694 five and five membrane spanning helices 656:694 The TMs consist of either five and five membrane spanning helices or five and six membrane spanning helices, depending upon the predictive algorithm used. 12600216 1 48 from homeostasis 152:162 arg1 photoreceptors 188:201 retinal rod and cone photoreceptors 167:201 retinal rod and cone photoreceptors 167:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 6 49 theme loops 924:928 arg1 orientation 837:847 the orientation 833:847 the orientation of the various membrane spanning helices 833:888 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 6 49 theme loops 924:928 arg1 localization 898:909 the localization 894:909 the localization of the short loops connecting these helices 894:953 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 0 50 theme exchanger 43:51 arg1 Topology 0:7 Topology 0:7 Topology of the retinal cone NCKX2 Na/Ca-K exchanger 0:51 Topology of the retinal cone NCKX2 Na/Ca-K exchanger. 12600216 7 51 theme extracellular 1039:1051 arg1 milieu 1053:1058 the extracellular milieu 1035:1058 the extracellular milieu 1035:1058 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 3 52 theme spanning 466:473 arg1 TMs 485:487 TMs 485:487 TMs 485:487 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 3 52 theme spanning 466:473 arg1 segments 475:482 transmembrane spanning segments 452:482 transmembrane spanning segments (TMs) 452:488 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 5 53 theme predictive 758:767 arg1 algorithm 769:777 the predictive algorithm 754:777 the predictive algorithm used 754:782 The TMs consist of either five and five membrane spanning helices or five and six membrane spanning helices, depending upon the predictive algorithm used. 12600216 0 54 theme Na/Ca-K 35:41 arg1 exchanger 43:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 the retinal cone NCKX2 Na/Ca-K exchanger 12:51 Topology of the retinal cone NCKX2 Na/Ca-K exchanger. 12600216 6 55 theme specific 792:799 arg1 information 801:811 Little specific information 785:811 Little specific information 785:811 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 3 56 theme Na/Ca-K 365:371 arg1 exchanger 373:381 the Na/Ca-K exchanger 361:381 the Na/Ca-K exchanger 361:381 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 3 57 theme segments 475:482 arg1 loops 430:434 two large hydrophilic loops 408:434 two large hydrophilic loops 408:434 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 3 57 theme segments 475:482 arg1 sets 444:447 two sets 440:447 two sets of transmembrane spanning segments (TMs) 440:488 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 3 57 theme segments 475:482 arg1 TMs 485:487 TMs 485:487 TMs 485:487 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 3 57 theme segments 475:482 arg1 segments 475:482 transmembrane spanning segments 452:482 transmembrane spanning segments (TMs) 452:488 The topology of the Na/Ca-K exchanger is thought to consist of two large hydrophilic loops and two sets of transmembrane spanning segments (TMs). 12600216 1 58 theme polytopic 88:96 arg1 protein 107:113 a polytopic membrane protein 86:113 a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors 86:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 1 58 theme polytopic 88:96 arg1 exchanger 66:74 The Na/Ca-K exchanger 54:74 The Na/Ca-K exchanger (NCKX) 54:81 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 8 59 theme spanning 1287:1294 arg1 helices 1296:1302 five membrane spanning helices 1273:1302 five membrane spanning helices 1273:1302 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 6 60 theme Little 785:790 arg1 information 801:811 Little specific information 785:811 Little specific information 785:811 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 2 61 theme NCKX1 208:212 arg1 isoform 214:220 The NCKX1 isoform 204:220 The NCKX1 isoform 204:220 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 6 62 theme helices 882:888 arg1 orientation 837:847 the orientation 833:847 the orientation of the various membrane spanning helices 833:888 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 6 62 theme helices 882:888 arg1 localization 898:909 the localization 894:909 the localization of the short loops connecting these helices 894:953 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 1 63 theme membrane 98:105 arg1 protein 107:113 a polytopic membrane protein 86:113 a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors 86:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 1 63 theme membrane 98:105 arg1 exchanger 66:74 The Na/Ca-K exchanger 54:74 The Na/Ca-K exchanger (NCKX) 54:81 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 1 64 theme retinal 167:173 arg1 photoreceptors 188:201 retinal rod and cone photoreceptors 167:201 retinal rod and cone photoreceptors 167:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 8 65 theme consistent 1206:1215 arg1 topology 1222:1229 a consistent NCKX topology 1204:1229 a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices 1204:1302 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 2 66 theme NCKX2 250:254 arg1 isoform 256:262 the NCKX2 isoform 246:262 the NCKX2 isoform 246:262 The NCKX1 isoform is found in rods, while the NCKX2 isoform is found in cones, in retinal ganglion cells, and in various parts of the brain. 12600216 1 67 theme rod 175:177 arg1 photoreceptors 188:201 retinal rod and cone photoreceptors 167:201 retinal rod and cone photoreceptors 167:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 12600216 7 68 theme different 1070:1078 arg1 insertion 1140:1148 insertion 1140:1148 insertion of N-glycosylation sites 1140:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 68 theme different 1070:1078 arg1 accessibility 1089:1101 accessibility 1089:1101 accessibility of substituted cysteine residues 1089:1134 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 68 theme different 1070:1078 arg1 methods 1080:1086 two different methods 1066:1086 two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites 1066:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 69 theme N-glycosylation 1153:1167 arg1 sites 1169:1173 N-glycosylation sites 1153:1173 N-glycosylation sites 1153:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 8 70 theme membrane 1278:1285 arg1 helices 1296:1302 five membrane spanning helices 1273:1302 five membrane spanning helices 1273:1302 The two methods resulted in a consistent NCKX topology in which the two sets of TMs each contain five membrane spanning helices. 12600216 7 71 theme sites 1169:1173 arg1 methods 1080:1086 two different methods 1066:1086 two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites 1066:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 71 theme sites 1169:1173 arg1 accessibility 1089:1101 accessibility 1089:1101 accessibility of substituted cysteine residues 1089:1134 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 7 71 theme sites 1169:1173 arg1 insertion 1140:1148 insertion 1140:1148 insertion of N-glycosylation sites 1140:1173 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 10 72 theme membrane 1575:1582 arg1 helices 1584:1590 the C-terminal three membrane helices 1554:1590 the C-terminal three membrane helices 1554:1590 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 9 73 from C-terminus 1409:1418 arg1 surface 1441:1447 the extracellular surface 1423:1447 the extracellular surface 1423:1447 Our new model places what was previously membrane spanning helix six in the cytoplasm, which places the C-terminus on the extracellular surface. 12600216 4 74 theme first 495:499 arg1 loop 519:522 The first large hydrophilic loop 491:522 The first large hydrophilic loop 491:522 The first large hydrophilic loop is located extracellularly at the N-terminus; the other is cytoplasmic and separates the two sets of TMs. 12600216 6 75 theme membrane 864:871 arg1 helices 882:888 the various membrane spanning helices 852:888 the various membrane spanning helices 852:888 Little specific information is yet available on the orientation of the various membrane spanning helices and the localization of the short loops connecting these helices. 12600216 7 76 theme substituted 1106:1116 arg1 residues 1127:1134 substituted cysteine residues 1106:1134 substituted cysteine residues 1106:1134 In this study, we have determined which of the connecting loops are exposed to the extracellular milieu using two different methods: accessibility of substituted cysteine residues and insertion of N-glycosylation sites. 12600216 10 77 theme NCKX 1469:1472 arg1 model 1483:1487 this NCKX topology model 1464:1487 this NCKX topology model 1464:1487 Surprisingly, this NCKX topology model is different from the current NCX topology model with respect to the C-terminal three membrane helices. 12600216 1 78 theme cone 183:186 arg1 photoreceptors 188:201 retinal rod and cone photoreceptors 167:201 retinal rod and cone photoreceptors 167:201 The Na/Ca-K exchanger (NCKX) is a polytopic membrane protein that plays a critical role in Ca(2+) homeostasis in retinal rod and cone photoreceptors. 7683678 9 0 theme family 1283:1288 arg1 member 1259:1264 a new member 1253:1264 a new member of the lipocalin family 1253:1288 It is thus a new member of the lipocalin family. 7683678 9 0 theme family 1283:1288 arg1 It 1242:1243 It 1242:1243 It 1242:1243 It is thus a new member of the lipocalin family. 7683678 4 1 theme 92-kDa 524:529 arg1 gelatinase 531:540 92-kDa gelatinase 524:540 92-kDa gelatinase 524:540 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 6 2 theme protein 907:913 arg1 protein 947:953 a 178-residue protein 933:953 a 178-residue protein 933:953 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 6 2 theme protein 907:913 arg1 structure 883:891 The primary structure 871:891 The primary structure of the 25-kDa protein 871:913 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 3 3 theme 25-kDa 330:335 arg1 protein 337:343 the 25-kDa protein 326:343 the 25-kDa protein 326:343 Specific antibodies against the 25-kDa protein were obtained by affinity purification of the gelatinase antibodies. 7683678 6 4 theme 178-residue 935:945 arg1 protein 947:953 a 178-residue protein 933:953 a 178-residue protein 933:953 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 6 4 theme 178-residue 935:945 arg1 structure 883:891 The primary structure 871:891 The primary structure of the 25-kDa protein 871:913 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 4 5 theme Immunoblotting 414:427 arg1 studies 453:459 Immunoblotting and immunoprecipitation studies 414:459 Immunoblotting and immunoprecipitation studies 414:459 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 4 6 theme 92-kDa 681:686 arg1 forms 688:692 the 220-, 135-, and 92-kDa forms 661:692 the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase 661:732 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 0 7 theme neutrophil 79:88 arg1 gelatinase 90:99 human neutrophil gelatinase 73:99 human neutrophil gelatinase 73:99 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 0 8 theme human 73:77 arg1 gelatinase 90:99 human neutrophil gelatinase 73:99 human neutrophil gelatinase 73:99 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 4 9 theme immunoprecipitation 433:451 arg1 studies 453:459 Immunoblotting and immunoprecipitation studies 414:459 Immunoblotting and immunoprecipitation studies 414:459 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 2 10 theme 25-kDa 282:287 arg1 protein 289:295 this 25-kDa protein 277:295 this 25-kDa protein 277:295 Polyclonal antibodies raised against gelatinase not only recognized gelatinase but also this 25-kDa protein. 7683678 5 11 theme 25-kDa 739:744 arg1 protein 746:752 The 25-kDa protein 735:752 The 25-kDa protein 735:752 The 25-kDa protein was purified to apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils. 7683678 8 12 theme 2-microglobulin-related 1182:1204 arg1 protein 1206:1212 rat alpha 2-microglobulin-related protein 1172:1212 rat alpha 2-microglobulin-related protein 1172:1212 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 4 13 theme 135- 671:674 arg1 forms 688:692 the 220-, 135-, and 92-kDa forms 661:692 the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase 661:732 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 4 14 theme 220- 665:668 arg1 forms 688:692 the 220-, 135-, and 92-kDa forms 661:692 the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase 661:732 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 8 15 theme alpha 1176:1180 arg1 protein 1206:1212 rat alpha 2-microglobulin-related protein 1172:1212 rat alpha 2-microglobulin-related protein 1172:1212 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 8 16 theme 24p3 1236:1239 arg1 sequences 1159:1167 the deduced sequences 1147:1167 the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3 1147:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 8 17 theme protein 1206:1212 arg1 sequences 1159:1167 the deduced sequences 1147:1167 the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3 1147:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 1 18 theme purified 151:158 arg1 gelatinase 177:186 purified human neutrophil gelatinase 151:186 purified human neutrophil gelatinase 151:186 A 25-kDa protein was found to be associated with purified human neutrophil gelatinase. 7683678 3 19 theme gelatinase 391:400 arg1 antibodies 402:411 the gelatinase antibodies 387:411 the gelatinase antibodies 387:411 Specific antibodies against the 25-kDa protein were obtained by affinity purification of the gelatinase antibodies. 7683678 5 20 theme apparent 770:777 arg1 homogeneity 779:789 apparent homogeneity 770:789 apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils 770:868 The 25-kDa protein was purified to apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils. 7683678 1 21 theme human 160:164 arg1 gelatinase 177:186 purified human neutrophil gelatinase 151:186 purified human neutrophil gelatinase 151:186 A 25-kDa protein was found to be associated with purified human neutrophil gelatinase. 7683678 3 22 theme antibodies 402:411 arg1 purification 371:382 affinity purification 362:382 affinity purification of the gelatinase antibodies 362:411 Specific antibodies against the 25-kDa protein were obtained by affinity purification of the gelatinase antibodies. 7683678 4 23 theme 220-kDa 574:580 arg1 form 582:585 the 220-kDa form 570:585 the 220-kDa form 570:585 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 4 23 theme 220-kDa 574:580 arg1 homodimer 612:620 a homodimer 610:620 a homodimer of the 92-kDa protein 610:642 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 8 24 theme human 1088:1092 arg1 protein 1094:1100 any known human protein 1078:1100 any known human protein 1078:1100 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 8 25 theme known 1082:1086 arg1 protein 1094:1100 any known human protein 1078:1100 any known human protein 1078:1100 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 10 26 theme protein 1318:1324 arg1 function 1295:1302 The function 1291:1302 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), 1291:1382 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), remains to be determined. 7683678 1 27 theme neutrophil 166:175 arg1 gelatinase 177:186 purified human neutrophil gelatinase 151:186 purified human neutrophil gelatinase 151:186 A 25-kDa protein was found to be associated with purified human neutrophil gelatinase. 7683678 8 28 theme mouse 1222:1226 arg1 24p3 1236:1239 the mouse protein 24p3 1218:1239 the mouse protein 24p3 1218:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 7 29 gly N-glycosylation 1014:1028 arg2 site 1030:1033 one N-glycosylation site 1010:1033 one N-glycosylation site 1010:1033 It was susceptible to treatment with N-glycanase, and one N-glycosylation site was identified. 7683678 7 29 gly N-glycosylation 1014:1028 arg2 one 1010:1012 one 1010:1012 one 1010:1012 It was susceptible to treatment with N-glycanase, and one N-glycosylation site was identified. 7683678 10 30 theme 25-kDa 1311:1316 arg1 protein 1318:1324 the 25-kDa protein 1307:1324 the 25-kDa protein 1307:1324 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), remains to be determined. 7683678 0 31 theme primary 14:20 arg1 structure 22:30 primary structure 14:30 primary structure 14:30 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 5 32 from neutrophils 858:868 arg1 material 807:814 exocytosed material 796:814 exocytosed material from phorbol myristate acetate-stimulated neutrophils 796:868 The 25-kDa protein was purified to apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils. 7683678 6 33 theme 25-kDa 900:905 arg1 protein 907:913 the 25-kDa protein 896:913 the 25-kDa protein 896:913 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 4 34 theme protein 636:642 arg1 form 582:585 the 220-kDa form 570:585 the 220-kDa form 570:585 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 4 34 theme protein 636:642 arg1 homodimer 612:620 a homodimer 610:620 a homodimer of the 92-kDa protein 610:642 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 0 35 theme NGAL 35:38 arg1 Isolation 0:8 Isolation 0:8 Isolation 0:8 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 0 35 theme NGAL 35:38 arg1 structure 22:30 primary structure 14:30 primary structure 14:30 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 0 35 theme NGAL 35:38 arg1 protein 49:55 a novel protein 41:55 a novel protein associated with human neutrophil gelatinase 41:99 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 4 36 theme characteristic 694:707 arg1 forms 688:692 the 220-, 135-, and 92-kDa forms 661:692 the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase 661:732 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 8 37 theme protein 1228:1234 arg1 24p3 1236:1239 the mouse protein 24p3 1218:1239 the mouse protein 24p3 1218:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 7 38 with treatment 978:986 arg1 N-glycanase 993:1003 N-glycanase 993:1003 N-glycanase 993:1003 It was susceptible to treatment with N-glycanase, and one N-glycosylation site was identified. 7683678 8 39 theme deduced 1151:1157 arg1 sequences 1159:1167 the deduced sequences 1147:1167 the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3 1147:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 10 40 theme gelatinase-associated 1344:1364 arg1 NGAL 1377:1380 NGAL 1377:1380 NGAL 1377:1380 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), remains to be determined. 7683678 10 40 theme gelatinase-associated 1344:1364 arg1 lipocalin 1366:1374 neutrophil gelatinase-associated lipocalin 1333:1374 neutrophil gelatinase-associated lipocalin (NGAL) 1333:1381 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), remains to be determined. 7683678 5 41 theme exocytosed 796:805 arg1 material 807:814 exocytosed material 796:814 exocytosed material from phorbol myristate acetate-stimulated neutrophils 796:868 The 25-kDa protein was purified to apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils. 7683678 3 42 theme affinity 362:369 arg1 purification 371:382 affinity purification 362:382 affinity purification of the gelatinase antibodies 362:411 Specific antibodies against the 25-kDa protein were obtained by affinity purification of the gelatinase antibodies. 7683678 4 43 theme gelatinase 494:503 arg1 form 486:489 the 135-kDa form 474:489 the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein 474:563 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 10 44 theme neutrophil 1333:1342 arg1 NGAL 1377:1380 NGAL 1377:1380 NGAL 1377:1380 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), remains to be determined. 7683678 10 44 theme neutrophil 1333:1342 arg1 lipocalin 1366:1374 neutrophil gelatinase-associated lipocalin 1333:1374 neutrophil gelatinase-associated lipocalin (NGAL) 1333:1381 The function of the 25-kDa protein, named neutrophil gelatinase-associated lipocalin (NGAL), remains to be determined. 7683678 8 45 theme high 1116:1119 arg1 degree 1121:1126 a high degree 1114:1126 a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3 1114:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 6 46 theme primary 875:881 arg1 protein 947:953 a 178-residue protein 933:953 a 178-residue protein 933:953 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 6 46 theme primary 875:881 arg1 structure 883:891 The primary structure 871:891 The primary structure of the 25-kDa protein 871:913 The primary structure of the 25-kDa protein was determined as a 178-residue protein. 7683678 9 47 theme new 1255:1257 arg1 member 1259:1264 a new member 1253:1264 a new member of the lipocalin family 1253:1288 It is thus a new member of the lipocalin family. 7683678 9 47 theme new 1255:1257 arg1 It 1242:1243 It 1242:1243 It 1242:1243 It is thus a new member of the lipocalin family. 7683678 5 48 from material 807:814 arg1 homogeneity 779:789 apparent homogeneity 770:789 apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils 770:868 The 25-kDa protein was purified to apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils. 7683678 1 49 theme 25-kDa 104:109 arg1 protein 111:117 A 25-kDa protein 102:117 A 25-kDa protein 102:117 A 25-kDa protein was found to be associated with purified human neutrophil gelatinase. 7683678 8 50 theme similarity 1131:1140 arg1 degree 1121:1126 a high degree 1114:1126 a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3 1114:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 2 51 theme Polyclonal 189:198 arg1 antibodies 200:209 Polyclonal antibodies 189:209 Polyclonal antibodies raised against gelatinase 189:235 Polyclonal antibodies raised against gelatinase not only recognized gelatinase but also this 25-kDa protein. 7683678 4 52 theme gelatinase 723:732 arg1 characteristic 694:707 characteristic 694:707 characteristic 694:707 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 8 53 theme rat 1172:1174 arg1 protein 1206:1212 rat alpha 2-microglobulin-related protein 1172:1212 rat alpha 2-microglobulin-related protein 1172:1212 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 3 54 theme Specific 298:305 arg1 antibodies 307:316 Specific antibodies 298:316 Specific antibodies against the 25-kDa protein 298:343 Specific antibodies against the 25-kDa protein were obtained by affinity purification of the gelatinase antibodies. 7683678 4 55 theme 135-kDa 478:484 arg1 form 486:489 the 135-kDa form 474:489 the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein 474:563 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 7 56 theme N-glycosylation 1014:1028 arg1 site 1030:1033 one N-glycosylation site 1010:1033 one N-glycosylation site 1010:1033 It was susceptible to treatment with N-glycanase, and one N-glycosylation site was identified. 7683678 4 57 theme neutrophil 712:721 arg1 gelatinase 723:732 neutrophil gelatinase 712:732 neutrophil gelatinase 712:732 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 9 58 theme lipocalin 1273:1281 arg1 family 1283:1288 the lipocalin family 1269:1288 the lipocalin family 1269:1288 It is thus a new member of the lipocalin family. 7683678 4 59 theme 92-kDa 629:634 arg1 protein 636:642 the 92-kDa protein 625:642 the 92-kDa protein 625:642 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 0 60 theme novel 43:47 arg1 Isolation 0:8 Isolation 0:8 Isolation 0:8 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 0 60 theme novel 43:47 arg1 protein 49:55 a novel protein 41:55 a novel protein associated with human neutrophil gelatinase 41:99 Isolation and primary structure of NGAL, a novel protein associated with human neutrophil gelatinase. 7683678 4 61 theme gelatinase 531:540 arg1 complex 513:519 a complex 511:519 a complex of 92-kDa gelatinase 511:540 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 4 61 theme gelatinase 531:540 arg1 protein 557:563 the 25-kDa protein 546:563 the 25-kDa protein 546:563 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 7683678 5 62 theme acetate-stimulated 839:856 arg1 neutrophils 858:868 phorbol myristate acetate-stimulated neutrophils 821:868 phorbol myristate acetate-stimulated neutrophils 821:868 The 25-kDa protein was purified to apparent homogeneity from exocytosed material from phorbol myristate acetate-stimulated neutrophils. 7683678 8 63 with similarity 1131:1140 arg1 sequences 1159:1167 the deduced sequences 1147:1167 the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3 1147:1239 The sequence did not match any known human protein, but showed a high degree of similarity with the deduced sequences of rat alpha 2-microglobulin-related protein and the mouse protein 24p3. 7683678 4 64 theme 25-kDa 550:555 arg1 protein 557:563 the 25-kDa protein 546:563 the 25-kDa protein 546:563 Immunoblotting and immunoprecipitation studies demonstrated the 135-kDa form of gelatinase to be a complex of 92-kDa gelatinase and the 25-kDa protein, and the 220-kDa form was demonstrated to be a homodimer of the 92-kDa protein, thus explaining the 220-, 135-, and 92-kDa forms characteristic of neutrophil gelatinase. 1694179 8 0 theme Timed 1040:1044 arg1 digests 1046:1052 Timed digests 1040:1052 Timed digests with N-glycosidase 1040:1071 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 12 1 theme protein 1842:1848 arg1 synthesis 1850:1858 protein synthesis 1842:1858 protein synthesis 1842:1858 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 4 2 theme numerous 507:514 arg1 inhibitors 527:536 numerous proteinase inhibitors 507:536 numerous proteinase inhibitors 507:536 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 7 3 theme synthetic 1007:1015 arg1 peptide 1031:1037 a synthetic COOH-terminal peptide 1005:1037 a synthetic COOH-terminal peptide 1005:1037 Both also react with antibodies to a synthetic COOH-terminal peptide. 1694179 10 4 theme serine 1439:1444 arg1 cysteine 1419:1426 cysteine 1419:1426 cysteine instead of serine or threonine 2 residues distal 1419:1475 This site is unique in having cysteine instead of serine or threonine 2 residues distal. 1694179 12 5 from related 1819:1825 arg1 turn 1811:1814 turn 1811:1814 turn 1811:1814 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 8 6 theme protein 1122:1128 arg1 C 1130:1130 beta protein C 1117:1130 beta protein C 1117:1130 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 12 7 from turn 1811:1814 arg1 related 1819:1825 related 1819:1825 related 1819:1825 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 8 8 contain has 1132:1134 arg2 chains 1167:1172 three N-linked oligosaccharide chains 1136:1172 three N-linked oligosaccharide chains 1136:1172 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 8 8 contain has 1132:1134 arg1 C 1130:1130 beta protein C 1117:1130 beta protein C 1117:1130 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 8 9 theme oligosaccharide 1151:1165 arg1 chains 1167:1172 three N-linked oligosaccharide chains 1136:1172 three N-linked oligosaccharide chains 1136:1172 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 4 10 theme protein 562:568 arg1 C 570:570 beta protein C 557:570 beta protein C 557:570 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 10 11 theme threonine 1449:1457 arg1 cysteine 1419:1426 cysteine 1419:1426 cysteine instead of serine or threonine 2 residues distal 1419:1475 This site is unique in having cysteine instead of serine or threonine 2 residues distal. 1694179 2 12 theme gel 271:273 arg1 electrophoresis 275:289 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 233:289 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 233:289 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 8 13 theme beta 1117:1120 arg1 C 1130:1130 beta protein C 1117:1130 beta protein C 1117:1130 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 8 14 theme protein 1188:1194 arg1 C 1196:1196 alpha protein C 1182:1196 alpha protein C 1182:1196 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 10 15 dep serine 1439:1444 arg1 residues 1461:1468 2 residues distal 1459:1475 2 residues distal 1459:1475 This site is unique in having cysteine instead of serine or threonine 2 residues distal. 1694179 12 16 theme bond 1783:1786 arg1 formation 1788:1796 disulfide bond formation 1773:1796 disulfide bond formation which may in turn be related to the rate of protein synthesis 1773:1858 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 4 17 theme protein 402:408 arg1 C 410:410 beta protein C 397:410 beta protein C 397:410 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 9 18 theme beta 1373:1376 arg1 C 1386:1386 beta protein C 1373:1386 beta protein C 1373:1386 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 6 19 theme chains 831:836 arg1 identical 856:864 identical 856:864 identical 856:864 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 6 19 theme chains 831:836 arg1 sequences 808:816 The NH2-terminal sequences 791:816 The NH2-terminal sequences of the heavy chains of both forms 791:850 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 8 20 with consistent 1077:1086 arg1 interpretation 1097:1110 the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four 1093:1205 the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four 1093:1205 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 11 21 theme glycosylation 1597:1609 arg1 reaction 1611:1618 the glycosylation reaction 1593:1618 the glycosylation reaction only until it forms a disulfide bond 1593:1655 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 1 22 theme usage 109:113 arg1 frequency 96:104 the frequency 92:104 the frequency of usage at asparagine-X-cysteine sites 92:144 The rate of translation may influence the frequency of usage at asparagine-X-cysteine sites. 1694179 2 23 theme predominant 203:213 arg1 form 215:218 the predominant form 199:218 the predominant form 199:218 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 1 24 theme asparagine-X-cysteine 118:138 arg1 sites 140:144 asparagine-X-cysteine sites 118:144 asparagine-X-cysteine sites 118:144 The rate of translation may influence the frequency of usage at asparagine-X-cysteine sites. 1694179 5 25 theme activated 683:691 arg1 X 700:700 activated factor X 683:700 activated factor X 683:700 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 1 26 from sites 140:144 arg1 frequency 96:104 the frequency 92:104 the frequency of usage at asparagine-X-cysteine sites 92:144 The rate of translation may influence the frequency of usage at asparagine-X-cysteine sites. 1694179 4 27 theme HepG2 442:446 arg1 line 453:456 the HepG2 cell line 438:456 the HepG2 cell line 438:456 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 9 28 gly glycosylated 1241:1252 arg2 329 1225:1227 asparagine 329 1214:1227 asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C 1214:1386 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 9 28 gly glycosylated 1241:1252 arg1 C 1270:1270 beta protein C 1257:1270 beta protein C 1257:1270 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 9 28 gly glycosylated 1241:1252 arg1 329 1225:1227 asparagine 329 1214:1227 asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C 1214:1386 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 5 29 theme protein 668:674 arg1 C 676:676 activated protein C 658:676 activated protein C 658:676 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 9 30 theme asparagine 1214:1223 arg1 329 1225:1227 asparagine 329 1214:1227 asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C 1214:1386 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 4 31 from present 465:471 arg1 plasma 476:481 plasma 476:481 plasma collected directly into numerous proteinase inhibitors 476:536 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 3 32 theme beta 348:351 arg1 C 361:361 beta protein C 348:361 beta protein C 348:361 It has been suggested that this species, referred to as beta protein C, is a degraded molecule. 1694179 3 33 theme degraded 369:376 arg1 species 324:330 this species 319:330 this species 319:330 It has been suggested that this species, referred to as beta protein C, is a degraded molecule. 1694179 3 33 theme degraded 369:376 arg1 molecule 378:385 a degraded molecule 367:385 a degraded molecule 367:385 It has been suggested that this species, referred to as beta protein C, is a degraded molecule. 1694179 0 34 theme Beta 0:3 arg1 C 13:13 Beta protein C 0:13 Beta protein C 0:13 Beta protein C is not glycosylated at asparagine 329. 1694179 1 35 theme translation 66:76 arg1 rate 58:61 The rate 54:61 The rate of translation 54:76 The rate of translation may influence the frequency of usage at asparagine-X-cysteine sites. 1694179 8 36 with digests 1046:1052 arg1 N-glycosidase 1059:1071 N-glycosidase 1059:1071 N-glycosidase 1059:1071 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 12 37 gly glycosylated 1694:1705 arg1 C 1684:1684 protein C 1676:1684 protein C that is glycosylated at this site 1676:1718 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 12 37 gly glycosylated 1694:1705 arg2 site 1715:1718 this site 1710:1718 this site 1710:1718 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 6 38 theme functional 921:930 arg1 site 939:942 a functional active site 919:942 a functional active site 919:942 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 11 39 theme hydroxyl 1546:1553 arg1 group 1555:1559 the usual hydroxyl group 1536:1559 the usual hydroxyl group 1536:1559 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 9 40 theme protein 1262:1268 arg1 C 1270:1270 beta protein C 1257:1270 beta protein C 1257:1270 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 12 41 theme C 1684:1684 arg1 percentage 1662:1671 The percentage 1658:1671 The percentage of protein C that is glycosylated at this site 1658:1718 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 12 42 theme protein 1676:1682 arg1 C 1684:1684 protein C 1676:1684 protein C that is glycosylated at this site 1676:1718 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 2 43 theme protein 173:179 arg1 C 181:181 human plasma protein C 160:181 human plasma protein C 160:181 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 5 44 dep presence 741:748 arg1 the 737:739 the 737:739 the 737:739 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 0 45 gly glycosylated 22:33 arg1 C 13:13 Beta protein C 0:13 Beta protein C 0:13 Beta protein C is not glycosylated at asparagine 329. 1694179 0 45 gly glycosylated 22:33 arg2 asparagine 38:47 asparagine 329 38:51 asparagine 329 38:51 Beta protein C is not glycosylated at asparagine 329. 1694179 11 46 theme hydrogen 1566:1573 arg1 acceptor 1580:1587 a hydrogen bond acceptor 1564:1587 a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond 1564:1655 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 6 47 theme heavy 825:829 arg1 chains 831:836 the heavy chains 821:836 the heavy chains of both forms 821:850 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 9 48 theme synthetic 1294:1302 arg1 peptide 1304:1310 a synthetic peptide 1292:1310 a synthetic peptide based on the sequence around this amino acid 1292:1355 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 2 49 theme sulfate-polyacrylamide 248:269 arg1 electrophoresis 275:289 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 233:289 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 233:289 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 6 50 theme NH2-terminal 795:806 arg1 identical 856:864 identical 856:864 identical 856:864 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 6 50 theme NH2-terminal 795:806 arg1 sequences 808:816 The NH2-terminal sequences 791:816 The NH2-terminal sequences of the heavy chains of both forms 791:850 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 4 51 theme proteinase 516:525 arg1 inhibitors 527:536 numerous proteinase inhibitors 507:536 numerous proteinase inhibitors 507:536 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 10 52 theme distal 1470:1475 arg1 residues 1461:1468 2 residues distal 1459:1475 2 residues distal 1459:1475 This site is unique in having cysteine instead of serine or threonine 2 residues distal. 1694179 5 53 theme calcium 764:770 arg1 presence 741:748 presence 741:748 presence 741:748 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 5 53 theme calcium 764:770 arg1 absence 753:759 absence 753:759 absence 753:759 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 12 54 theme formation 1788:1796 arg1 rate 1765:1768 the rate 1761:1768 the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis 1761:1858 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 2 55 theme C 181:181 arg1 % 155:155 About 30% 147:155 About 30% of human plasma protein C 147:181 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 2 55 theme C 181:181 arg1 C 181:181 human plasma protein C 160:181 human plasma protein C 160:181 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 8 56 theme N-linked 1142:1149 arg1 chains 1167:1172 three N-linked oligosaccharide chains 1136:1172 three N-linked oligosaccharide chains 1136:1172 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 9 57 theme amino 1346:1350 arg1 acid 1352:1355 this amino acid 1341:1355 this amino acid 1341:1355 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 4 58 theme beta 397:400 arg1 C 410:410 beta protein C 397:410 beta protein C 397:410 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 4 59 theme beta 557:560 arg1 C 570:570 beta protein C 557:570 beta protein C 557:570 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 11 60 theme disulfide 1642:1650 arg1 bond 1652:1655 a disulfide bond 1640:1655 a disulfide bond 1640:1655 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 6 61 theme forms 846:850 arg1 chains 831:836 the heavy chains 821:836 the heavy chains of both forms 821:850 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 4 62 theme C 570:570 arg1 percentage 543:552 the percentage 539:552 the percentage of beta protein C 539:570 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 12 63 theme disulfide 1773:1781 arg1 formation 1788:1796 disulfide bond formation 1773:1796 disulfide bond formation which may in turn be related to the rate of protein synthesis 1773:1858 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 4 64 theme blood 622:626 arg1 collection 628:637 blood collection 622:637 blood collection 622:637 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 9 65 theme protein 1378:1384 arg1 C 1386:1386 beta protein C 1373:1386 beta protein C 1373:1386 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 8 66 contain has 1198:1200 arg1 C 1196:1196 alpha protein C 1182:1196 alpha protein C 1182:1196 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 8 66 contain has 1198:1200 arg2 four 1202:1205 four 1202:1205 four 1202:1205 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 5 67 theme phospholipids 776:788 arg1 presence 741:748 presence 741:748 presence 741:748 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 5 67 theme phospholipids 776:788 arg1 absence 753:759 absence 753:759 absence 753:759 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 12 68 theme synthesis 1850:1858 arg1 rate 1834:1837 the rate 1830:1837 the rate of protein synthesis 1830:1858 The percentage of protein C that is glycosylated at this site may therefore depend at least in part on the rate of disulfide bond formation which may in turn be related to the rate of protein synthesis. 1694179 6 69 theme same 888:891 arg1 dodecapeptide 893:905 the same dodecapeptide 884:905 the same dodecapeptide 884:905 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 4 70 theme cell 448:451 arg1 line 453:456 the HepG2 cell line 438:456 the HepG2 cell line 438:456 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 4 71 attach present 465:471 arg2 C 410:410 beta protein C 397:410 beta protein C 397:410 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 4 71 attach present 465:471 arg1 plasma 476:481 plasma 476:481 plasma collected directly into numerous proteinase inhibitors 476:536 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 5 72 theme activated 658:666 arg1 C 676:676 activated protein C 658:676 activated protein C 658:676 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 5 73 theme alpha 715:719 arg1 form 721:724 the alpha form 711:724 the alpha form to beta 711:732 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 3 74 theme protein 353:359 arg1 C 361:361 beta protein C 348:361 beta protein C 348:361 It has been suggested that this species, referred to as beta protein C, is a degraded molecule. 1694179 2 75 theme human 160:164 arg1 C 181:181 human plasma protein C 160:181 human plasma protein C 160:181 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 4 76 from plasma 476:481 arg1 present 465:471 present 465:471 present 465:471 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 4 77 located present 465:471 arg2 C 410:410 beta protein C 397:410 beta protein C 397:410 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 4 77 located present 465:471 arg1 plasma 476:481 plasma 476:481 plasma collected directly into numerous proteinase inhibitors 476:536 However, beta protein C is secreted in culture by the HepG2 cell line and is present in plasma collected directly into numerous proteinase inhibitors; the percentage of beta protein C does not change with time during culture or after blood collection. 1694179 5 78 theme factor 693:698 arg1 X 700:700 activated factor X 683:700 activated factor X 683:700 Neither thrombin, activated protein C, nor activated factor X converts the alpha form to beta in the presence or absence of calcium and phospholipids. 1694179 8 79 link N-linked 1142:1149 arg1 chains 1167:1172 three N-linked oligosaccharide chains 1136:1172 three N-linked oligosaccharide chains 1136:1172 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 6 80 theme active 932:937 arg1 site 939:942 a functional active site 919:942 a functional active site 919:942 The NH2-terminal sequences of the heavy chains of both forms are identical, and both release the same dodecapeptide and develop a functional active site when cleaved by thrombin. 1694179 11 81 theme usual 1540:1544 arg1 group 1555:1559 the usual hydroxyl group 1536:1559 the usual hydroxyl group 1536:1559 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 7 82 theme COOH-terminal 1017:1029 arg1 peptide 1031:1037 a synthetic COOH-terminal peptide 1005:1037 a synthetic COOH-terminal peptide 1005:1037 Both also react with antibodies to a synthetic COOH-terminal peptide. 1694179 9 83 theme beta 1257:1260 arg1 C 1270:1270 beta protein C 1257:1270 beta protein C 1257:1270 It is asparagine 329 that is not glycosylated in beta protein C since antibodies to a synthetic peptide based on the sequence around this amino acid react only with beta protein C. 1694179 11 84 theme bond 1575:1578 arg1 acceptor 1580:1587 a hydrogen bond acceptor 1564:1587 a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond 1564:1655 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 2 85 theme plasma 166:171 arg1 C 181:181 human plasma protein C 160:181 human plasma protein C 160:181 About 30% of human plasma protein C is smaller than the predominant form as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1694179 8 86 theme alpha 1182:1186 arg1 C 1196:1196 alpha protein C 1182:1196 alpha protein C 1182:1196 Timed digests with N-glycosidase are consistent with the interpretation that beta protein C has three N-linked oligosaccharide chains whereas alpha protein C has four. 1694179 11 87 theme sulfhydryl 1500:1509 arg1 group 1511:1515 the sulfhydryl group 1496:1515 the sulfhydryl group 1496:1515 It is likely that the sulfhydryl group can substitute for the usual hydroxyl group as a hydrogen bond acceptor for the glycosylation reaction only until it forms a disulfide bond. 1694179 0 88 theme protein 5:11 arg1 C 13:13 Beta protein C 0:13 Beta protein C 0:13 Beta protein C is not glycosylated at asparagine 329. 1693148 11 0 theme purified 1535:1542 arg1 IFN-omega 1544:1552 purified IFN-omega 1 1535:1554 purified IFN-omega 1 1535:1554 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 3 1 theme Sendai 477:482 arg1 virus 484:488 Sendai virus 477:488 Sendai virus 477:488 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 3 2 attach derived 423:429 arg1 leukocytes 453:462 peripheral blood leukocytes 436:462 peripheral blood leukocytes induced with Sendai virus 436:488 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 3 2 attach derived 423:429 arg2 IFN 418:420 human leukocyte IFN 402:420 human leukocyte IFN 402:420 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 4 3 theme resulting 568:576 arg1 pure 603:606 pure 603:606 pure 603:606 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 4 3 theme resulting 568:576 arg1 protein 578:584 The resulting protein 564:584 The resulting protein 564:584 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 9 4 contain containing 1379:1388 arg1 oligosaccharides 1362:1377 biantennary complex oligosaccharides 1342:1377 biantennary complex oligosaccharides containing neuraminic acid 1342:1404 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 9 4 contain containing 1379:1388 arg2 acid 1401:1404 neuraminic acid 1390:1404 neuraminic acid 1390:1404 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 5 5 theme NH2 863:865 arg1 terminus 867:874 the NH2 terminus 859:874 the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha 859:936 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 8 6 theme enzymatic 1205:1213 arg1 deglycosylation 1215:1229 enzymatic deglycosylation 1205:1229 enzymatic deglycosylation 1205:1229 Essentially all IFN-omega 1 molecules are glycosylated; enzymatic deglycosylation resulted in a reduction of the Mr to 20,500. 1693148 4 7 theme high 639:642 arg1 chromatography 663:676 reverse-phase high performance liquid chromatography 625:676 reverse-phase high performance liquid chromatography 625:676 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 2 8 theme IFN-omega 290:298 arg1 activities 276:285 the biological activities 261:285 the biological activities of IFN-omega 1 and its physiological role 261:327 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 8 9 theme IFN-omega 1165:1173 arg1 molecules 1177:1185 Essentially all IFN-omega 1 molecules 1149:1185 Essentially all IFN-omega 1 molecules 1149:1185 Essentially all IFN-omega 1 molecules are glycosylated; enzymatic deglycosylation resulted in a reduction of the Mr to 20,500. 1693148 11 10 dep 2.7 1600:1602 arg1 8 1609:1609 8 1609:1609 8 1609:1609 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 11 theme bovine 1720:1725 arg1 cells 1711:1715 cells 1711:1715 cells of bovine and ovine but not of equine or murine origin 1711:1770 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 3 12 theme monoclonal 519:528 arg1 chromatography 548:561 monoclonal antibody affinity chromatography 519:561 monoclonal antibody affinity chromatography 519:561 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 11 13 theme equine 1748:1753 arg1 origin 1765:1770 equine or murine origin 1748:1770 equine or murine origin 1748:1770 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 7 14 theme COOH-terminal 1110:1122 arg1 heterogeneity 1124:1136 COOH-terminal heterogeneity 1110:1136 COOH-terminal heterogeneity 1110:1136 No evidence for COOH-terminal heterogeneity was found. 1693148 8 15 theme Mr 1262:1263 arg1 reduction 1245:1253 a reduction 1243:1253 a reduction of the Mr to 20,500 1243:1273 Essentially all IFN-omega 1 molecules are glycosylated; enzymatic deglycosylation resulted in a reduction of the Mr to 20,500. 1693148 3 16 theme sequential 498:507 arg1 cycles 509:514 two sequential cycles 494:514 two sequential cycles of monoclonal antibody affinity chromatography 494:561 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 5 17 contain have 854:857 arg2 terminus 867:874 the NH2 terminus 859:874 the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha 859:936 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 5 17 contain have 854:857 arg1 % 835:835 only about 40% 822:835 only about 40% of the molecules 822:852 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 5 17 contain have 854:857 arg1 molecules 844:852 the molecules 840:852 the molecules 840:852 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 4 18 theme liquid 656:661 arg1 chromatography 663:676 reverse-phase high performance liquid chromatography 625:676 reverse-phase high performance liquid chromatography 625:676 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 11 19 theme antiviral 1671:1679 arg1 activity 1681:1688 potent antiviral activity 1664:1688 potent antiviral activity 1664:1688 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 2 20 theme related 229:235 arg1 protein 208:214 a recently discovered protein 186:214 a recently discovered protein structurally related to IFN-alpha and -beta 186:258 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 20 theme related 229:235 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 5 21 theme molecules 844:852 arg1 % 835:835 only about 40% 822:835 only about 40% of the molecules 822:852 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 5 21 theme molecules 844:852 arg1 molecules 844:852 the molecules 840:852 the molecules 840:852 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 11 22 theme antiviral 1513:1521 arg1 activity 1523:1530 The specific antiviral activity 1500:1530 The specific antiviral activity of purified IFN-omega 1 assayed on human cells 1500:1577 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 22 theme antiviral 1513:1521 arg1 IU/mg 1612:1616 2.7 x 10(8) IU/mg 1600:1616 2.7 x 10(8) IU/mg 1600:1616 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 2 23 theme IFN-alpha 168:176 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 23 theme IFN-alpha 168:176 arg1 II1 178:180 IFN-omega 1 = IFN-alpha II1 154:180 IFN-omega 1 = IFN-alpha II1 154:180 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 5 24 theme similarity 914:923 arg1 basis 892:896 the basis 888:896 the basis of the sequence similarity to IFN-alpha 888:936 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 4 25 theme theoretical 754:764 arg1 19,984 770:775 19,984 770:775 19,984 770:775 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 4 25 theme theoretical 754:764 arg1 Mr 766:767 theoretical Mr 754:767 theoretical Mr 754:767 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 3 26 theme human 402:406 arg1 IFN 418:420 human leukocyte IFN 402:420 human leukocyte IFN 402:420 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 1 27 theme alternative 75:85 arg1 sites 96:100 Two alternative cleavage sites 71:100 Two alternative cleavage sites for the signal peptidase 71:125 Two alternative cleavage sites for the signal peptidase. 1693148 5 28 theme Amino 779:783 arg1 sequence 790:797 Amino acid sequence 779:797 Amino acid sequence analysis 779:806 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 9 29 theme plant 1302:1306 arg1 lectins 1308:1314 several plant lectins 1294:1314 several plant lectins 1294:1314 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 11 30 theme class 1650:1654 arg1 IFNs 1658:1661 other human class I IFNs 1638:1661 other human class I IFNs 1638:1661 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 10 31 theme 8.5 1495:1497 arg1 points 1477:1482 isoelectric points 1465:1482 isoelectric points of 8.1 and 8.5 1465:1497 Two major peaks were observed upon chromatofocusing, with isoelectric points of 8.1 and 8.5. 1693148 4 32 theme dodecyl 717:723 arg1 electrophoresis 737:751 sodium dodecyl sulfate-gel electrophoresis 710:751 sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984) 710:776 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 5 33 theme sequence 790:797 arg1 analysis 799:806 Amino acid sequence analysis 779:806 Amino acid sequence analysis 779:806 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 10 34 theme 8.1 1487:1489 arg1 points 1477:1482 isoelectric points 1465:1482 isoelectric points of 8.1 and 8.5 1465:1497 Two major peaks were observed upon chromatofocusing, with isoelectric points of 8.1 and 8.5. 1693148 2 35 theme = 166:166 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 35 theme = 166:166 arg1 II1 178:180 IFN-omega 1 = IFN-alpha II1 154:180 IFN-omega 1 = IFN-alpha II1 154:180 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 3 36 theme IFN 418:420 arg1 preparations 386:397 preparations 386:397 preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, 386:489 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 0 37 theme natural 37:43 arg1 omega 62:66 natural human interferon omega 1 37:68 natural human interferon omega 1 37:68 Purification and characterization of natural human interferon omega 1. 1693148 2 38 theme IFN-omega 154:162 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 38 theme IFN-omega 154:162 arg1 II1 178:180 IFN-omega 1 = IFN-alpha II1 154:180 IFN-omega 1 = IFN-alpha II1 154:180 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 39 theme role 324:327 arg1 activities 276:285 the biological activities 261:285 the biological activities of IFN-omega 1 and its physiological role 261:327 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 0 40 theme interferon 51:60 arg1 omega 62:66 natural human interferon omega 1 37:68 natural human interferon omega 1 37:68 Purification and characterization of natural human interferon omega 1. 1693148 9 41 theme biantennary 1342:1352 arg1 oligosaccharides 1362:1377 biantennary complex oligosaccharides 1342:1377 biantennary complex oligosaccharides containing neuraminic acid 1342:1404 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 5 42 theme amino 981:985 arg1 acids 987:991 two additional amino acids 966:991 two additional amino acids 966:991 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 6 43 theme signal 1076:1081 arg1 peptidase 1083:1091 the signal peptidase 1072:1091 the signal peptidase 1072:1091 This difference probably results from variable cleavage of the pre-protein by the signal peptidase. 1693148 11 44 theme potent 1664:1669 arg1 activity 1681:1688 potent antiviral activity 1664:1688 potent antiviral activity 1664:1688 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 45 located observed 1699:1706 arg1 cells 1711:1715 cells 1711:1715 cells of bovine and ovine but not of equine or murine origin 1711:1770 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 45 located observed 1699:1706 arg2 activity 1681:1688 potent antiviral activity 1664:1688 potent antiviral activity 1664:1688 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 3 46 theme peripheral 436:445 arg1 leukocytes 453:462 peripheral blood leukocytes 436:462 peripheral blood leukocytes induced with Sendai virus 436:488 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 9 47 theme oligosaccharides 1362:1377 arg1 presence 1330:1337 the presence 1326:1337 the presence of biantennary complex oligosaccharides containing neuraminic acid 1326:1404 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 4 48 theme performance 644:654 arg1 chromatography 663:676 reverse-phase high performance liquid chromatography 625:676 reverse-phase high performance liquid chromatography 625:676 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 9 49 theme neuraminic 1390:1399 arg1 acid 1401:1404 neuraminic acid 1390:1404 neuraminic acid 1390:1404 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 11 50 theme similar 1619:1625 arg1 activity 1523:1530 The specific antiviral activity 1500:1530 The specific antiviral activity of purified IFN-omega 1 assayed on human cells 1500:1577 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 50 theme similar 1619:1625 arg1 IU/mg 1612:1616 2.7 x 10(8) IU/mg 1600:1616 2.7 x 10(8) IU/mg 1600:1616 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 51 theme other 1638:1642 arg1 IFNs 1658:1661 other human class I IFNs 1638:1661 other human class I IFNs 1638:1661 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 52 theme human 1567:1571 arg1 cells 1573:1577 human cells 1567:1577 human cells 1567:1577 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 53 theme IFN-omega 1544:1552 arg1 activity 1523:1530 The specific antiviral activity 1500:1530 The specific antiviral activity of purified IFN-omega 1 assayed on human cells 1500:1577 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 53 theme IFN-omega 1544:1552 arg1 IU/mg 1612:1616 2.7 x 10(8) IU/mg 1600:1616 2.7 x 10(8) IU/mg 1600:1616 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 2 54 theme biological 265:274 arg1 activities 276:285 the biological activities 261:285 the biological activities of IFN-omega 1 and its physiological role 261:327 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 55 theme interferon 134:143 arg1 protein 208:214 a recently discovered protein 186:214 a recently discovered protein structurally related to IFN-alpha and -beta 186:258 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 55 theme interferon 134:143 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 55 theme interferon 134:143 arg1 II1 178:180 IFN-omega 1 = IFN-alpha II1 154:180 IFN-omega 1 = IFN-alpha II1 154:180 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 10 56 theme major 1411:1415 arg1 peaks 1417:1421 Two major peaks 1407:1421 Two major peaks 1407:1421 Two major peaks were observed upon chromatofocusing, with isoelectric points of 8.1 and 8.5. 1693148 3 57 theme antibody 530:537 arg1 chromatography 548:561 monoclonal antibody affinity chromatography 519:561 monoclonal antibody affinity chromatography 519:561 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 4 58 theme reverse-phase 625:637 arg1 chromatography 663:676 reverse-phase high performance liquid chromatography 625:676 reverse-phase high performance liquid chromatography 625:676 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 8 59 gly glycosylated 1191:1202 arg1 molecules 1177:1185 Essentially all IFN-omega 1 molecules 1149:1185 Essentially all IFN-omega 1 molecules 1149:1185 Essentially all IFN-omega 1 molecules are glycosylated; enzymatic deglycosylation resulted in a reduction of the Mr to 20,500. 1693148 8 60 theme all 1161:1163 arg1 molecules 1177:1185 Essentially all IFN-omega 1 molecules 1149:1185 Essentially all IFN-omega 1 molecules 1149:1185 Essentially all IFN-omega 1 molecules are glycosylated; enzymatic deglycosylation resulted in a reduction of the Mr to 20,500. 1693148 3 61 theme chromatography 548:561 arg1 cycles 509:514 two sequential cycles 494:514 two sequential cycles of monoclonal antibody affinity chromatography 494:561 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 1 62 theme signal 110:115 arg1 peptidase 117:125 the signal peptidase 106:125 the signal peptidase 106:125 Two alternative cleavage sites for the signal peptidase. 1693148 11 63 theme murine 1758:1763 arg1 origin 1765:1770 equine or murine origin 1748:1770 equine or murine origin 1748:1770 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 3 64 theme affinity 539:546 arg1 chromatography 548:561 monoclonal antibody affinity chromatography 519:561 monoclonal antibody affinity chromatography 519:561 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 11 65 theme specific 1504:1511 arg1 activity 1523:1530 The specific antiviral activity 1500:1530 The specific antiviral activity of purified IFN-omega 1 assayed on human cells 1500:1577 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 65 theme specific 1504:1511 arg1 IU/mg 1612:1616 2.7 x 10(8) IU/mg 1600:1616 2.7 x 10(8) IU/mg 1600:1616 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 4 66 dep electrophoresis 737:751 arg1 19,984 770:775 19,984 770:775 19,984 770:775 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 4 66 dep electrophoresis 737:751 arg1 Mr 766:767 theoretical Mr 754:767 theoretical Mr 754:767 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 5 67 theme sequence 905:912 arg1 similarity 914:923 the sequence similarity 901:923 the sequence similarity to IFN-alpha 901:936 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 4 68 theme sulfate-gel 725:735 arg1 electrophoresis 737:751 sodium dodecyl sulfate-gel electrophoresis 710:751 sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984) 710:776 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 3 69 theme blood 447:451 arg1 leukocytes 453:462 peripheral blood leukocytes 436:462 peripheral blood leukocytes induced with Sendai virus 436:488 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 6 70 theme variable 1032:1039 arg1 cleavage 1041:1048 variable cleavage 1032:1048 variable cleavage of the pre-protein by the signal peptidase 1032:1091 This difference probably results from variable cleavage of the pre-protein by the signal peptidase. 1693148 10 71 theme isoelectric 1465:1475 arg1 points 1477:1482 isoelectric points 1465:1482 isoelectric points of 8.1 and 8.5 1465:1497 Two major peaks were observed upon chromatofocusing, with isoelectric points of 8.1 and 8.5. 1693148 3 72 theme leukocyte 408:416 arg1 IFN 418:420 human leukocyte IFN 402:420 human leukocyte IFN 402:420 We have purified IFN-omega 1 from preparations of human leukocyte IFN, derived from peripheral blood leukocytes induced with Sendai virus, by two sequential cycles of monoclonal antibody affinity chromatography. 1693148 9 73 theme several 1294:1300 arg1 lectins 1308:1314 several plant lectins 1294:1314 several plant lectins 1294:1314 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 1 74 theme cleavage 87:94 arg1 sites 96:100 Two alternative cleavage sites 71:100 Two alternative cleavage sites for the signal peptidase 71:125 Two alternative cleavage sites for the signal peptidase. 1693148 2 75 theme discovered 197:206 arg1 protein 208:214 a recently discovered protein 186:214 a recently discovered protein structurally related to IFN-alpha and -beta 186:258 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 75 theme discovered 197:206 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 4 76 theme 24,500 698:703 arg1 Mr 692:693 an Mr 689:693 an Mr of 24,500 689:703 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 5 77 theme acid 785:788 arg1 sequence 790:797 Amino acid sequence 779:797 Amino acid sequence analysis 779:806 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 11 78 theme human 1644:1648 arg1 IFNs 1658:1661 other human class I IFNs 1638:1661 other human class I IFNs 1638:1661 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 4 79 theme sodium 710:715 arg1 electrophoresis 737:751 sodium dodecyl sulfate-gel electrophoresis 710:751 sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984) 710:776 The resulting protein was at least 95% pure as determined by reverse-phase high performance liquid chromatography and showed an Mr of 24,500 upon sodium dodecyl sulfate-gel electrophoresis (theoretical Mr, 19,984). 1693148 0 80 theme human 45:49 arg1 omega 62:66 natural human interferon omega 1 37:68 natural human interferon omega 1 37:68 Purification and characterization of natural human interferon omega 1. 1693148 0 81 theme omega 62:66 arg1 characterization 17:32 characterization 17:32 characterization 17:32 Purification and characterization of natural human interferon omega 1. 1693148 0 81 theme omega 62:66 arg1 Purification 0:11 Purification 0:11 Purification 0:11 Purification and characterization of natural human interferon omega 1. 1693148 5 82 theme additional 970:979 arg1 acids 987:991 two additional amino acids 966:991 two additional amino acids 966:991 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 2 83 theme physiological 310:322 arg1 role 324:327 its physiological role 306:327 its physiological role 306:327 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 9 84 theme complex 1354:1360 arg1 oligosaccharides 1362:1377 biantennary complex oligosaccharides 1342:1377 biantennary complex oligosaccharides containing neuraminic acid 1342:1404 Experiments using several plant lectins indicated the presence of biantennary complex oligosaccharides containing neuraminic acid. 1693148 11 85 theme I 1656:1656 arg1 IFNs 1658:1661 other human class I IFNs 1638:1661 other human class I IFNs 1638:1661 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 11 86 theme origin 1765:1770 arg1 cells 1711:1715 cells 1711:1715 cells of bovine and ovine but not of equine or murine origin 1711:1770 The specific antiviral activity of purified IFN-omega 1 assayed on human cells was determined to be 2.7 x 10(8) IU/mg, similar to that of other human class I IFNs; potent antiviral activity was also observed on cells of bovine and ovine but not of equine or murine origin. 1693148 6 87 theme pre-protein 1057:1067 arg1 cleavage 1041:1048 variable cleavage 1032:1048 variable cleavage of the pre-protein by the signal peptidase 1032:1091 This difference probably results from variable cleavage of the pre-protein by the signal peptidase. 1693148 5 88 contain contain 958:964 arg2 acids 987:991 two additional amino acids 966:991 two additional amino acids 966:991 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 5 88 contain contain 958:964 arg1 others 951:956 the others 947:956 the others 947:956 Amino acid sequence analysis revealed that only about 40% of the molecules have the NH2 terminus expected on the basis of the sequence similarity to IFN-alpha, whereas the others contain two additional amino acids. 1693148 2 89 theme Human 128:132 arg1 protein 208:214 a recently discovered protein 186:214 a recently discovered protein structurally related to IFN-alpha and -beta 186:258 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 89 theme Human 128:132 arg1 omega 145:149 Human interferon omega 1 128:151 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) 128:181 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 1693148 2 89 theme Human 128:132 arg1 II1 178:180 IFN-omega 1 = IFN-alpha II1 154:180 IFN-omega 1 = IFN-alpha II1 154:180 Human interferon omega 1 (IFN-omega 1 = IFN-alpha II1) is a recently discovered protein structurally related to IFN-alpha and -beta; the biological activities of IFN-omega 1 and its physiological role are not known to date. 22023369 1 0 theme Fcγ 312:314 arg1 FcγRIIIa 331:338 FcγRIIIa 331:338 FcγRIIIa 331:338 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 0 theme Fcγ 312:314 arg1 IIIa 325:328 Fcγ receptor IIIa 312:328 Fcγ receptor IIIa (FcγRIIIa) 312:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 4 1 theme receptor 814:821 arg1 site 831:834 the receptor binding site 810:834 the receptor binding site 810:834 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 1 2 theme dramatic 210:217 arg1 enhancement 219:229 a dramatic enhancement 208:229 a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa) 208:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 2 3 theme soluble 438:444 arg1 sFcγRIIIa 464:472 sFcγRIIIa 464:472 sFcγRIIIa 464:472 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 2 3 theme soluble 438:444 arg1 form 446:449 a soluble form 436:449 a soluble form of FcγRIIIa (sFcγRIIIa) 436:473 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 1 4 theme receptor 316:323 arg1 FcγRIIIa 331:338 FcγRIIIa 331:338 FcγRIIIa 331:338 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 4 theme receptor 316:323 arg1 IIIa 325:328 Fcγ receptor IIIa 312:328 Fcγ receptor IIIa (FcγRIIIa) 312:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 5 from N-glycans 143:151 arg1 Removal 104:110 Removal 104:110 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) 104:195 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 2 6 gly N-glycosylation 484:498 arg2 two 480:482 two 480:482 two 480:482 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 2 6 gly N-glycosylation 484:498 arg2 sites 500:504 two N-glycosylation sites 480:504 two N-glycosylation sites 480:504 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 0 7 theme glycans 95:101 arg1 defucosylation 68:81 defucosylation 68:81 defucosylation of their Fc glycans 68:101 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 4 8 theme site 831:834 arg1 dynamics 798:805 the dynamics 794:805 the dynamics of the receptor binding site 794:834 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 5 9 theme therapeutic 901:911 arg1 antibodies 913:922 therapeutic antibodies 901:922 therapeutic antibodies 901:922 Our results offer a structural basis for improvement in ADCC of therapeutic antibodies by defucosylation. 22023369 4 10 theme N-glycans 692:700 arg1 fucosylation 669:680 fucosylation 669:680 fucosylation of the Fc N-glycans 669:700 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 0 11 theme Fc 92:93 arg1 glycans 95:101 their Fc glycans 86:101 their Fc glycans 86:101 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 4 12 theme binding 823:829 arg1 site 831:834 the receptor binding site 810:834 the receptor binding site 810:834 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 0 13 gly defucosylation 68:81 arg1 glycans 95:101 their Fc glycans 86:101 their Fc glycans 86:101 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 1 14 theme antibody-dependent 234:251 arg1 ADCC 276:279 ADCC 276:279 ADCC 276:279 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 14 theme antibody-dependent 234:251 arg1 cytotoxicity 262:273 antibody-dependent cellular cytotoxicity 234:273 antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa) 234:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 4 15 gly fucosylation 669:680 arg1 N-glycans 692:700 the Fc N-glycans 685:700 the Fc N-glycans 685:700 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 4 15 gly fucosylation 669:680 arg1 site 831:834 the receptor binding site 810:834 the receptor binding site 810:834 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 2 16 theme nonfucosylated 409:422 arg1 IgG1-Fc 424:430 nonfucosylated IgG1-Fc 409:430 nonfucosylated IgG1-Fc 409:430 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 1 17 theme Fc 160:161 arg1 portion 163:169 the Fc portion 156:169 the Fc portion of immunoglobulin G (IgG) 156:195 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 0 18 from efficacy 30:37 arg1 defucosylation 68:81 defucosylation 68:81 defucosylation of their Fc glycans 68:101 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 4 19 theme Fc 689:690 arg1 N-glycans 692:700 the Fc N-glycans 685:700 the Fc N-glycans 685:700 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 1 20 theme portion 163:169 arg1 N-glycans 143:151 the N-glycans 139:151 the N-glycans of the Fc portion of immunoglobulin G (IgG) 139:195 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 0 21 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 0:102 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 1 22 theme cellular 253:260 arg1 ADCC 276:279 ADCC 276:279 ADCC 276:279 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 22 theme cellular 253:260 arg1 cytotoxicity 262:273 antibody-dependent cellular cytotoxicity 234:273 antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa) 234:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 0 23 theme improved 21:28 arg1 efficacy 30:37 improved efficacy 21:37 improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans 21:101 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 1 24 theme cytotoxicity 262:273 arg1 enhancement 219:229 a dramatic enhancement 208:229 a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa) 208:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 25 theme immunoglobulin 174:187 arg1 G 189:189 immunoglobulin G 174:189 immunoglobulin G (IgG) 174:195 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 1 25 theme immunoglobulin 174:187 arg1 IgG 192:194 IgG 192:194 IgG 192:194 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 3 26 theme crystal 511:517 arg1 structure 519:527 The crystal structure 507:527 The crystal structure 507:527 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 22023369 3 27 with interaction 591:601 arg1 Fc 623:624 nonfucosylated Fc 608:624 nonfucosylated Fc 608:624 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 22023369 1 28 theme G 189:189 arg1 portion 163:169 the Fc portion 156:169 the Fc portion of immunoglobulin G (IgG) 156:195 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 2 29 theme N-glycosylation 484:498 arg1 sites 500:504 two N-glycosylation sites 480:504 two N-glycosylation sites 480:504 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 2 30 theme 2.2-Å 363:367 arg1 structure 369:377 the 2.2-Å structure 359:377 the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites 359:504 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 2 31 theme complex 386:392 arg1 structure 369:377 the 2.2-Å structure 359:377 the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites 359:504 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 0 32 theme antibodies 54:63 arg1 efficacy 30:37 improved efficacy 21:37 improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans 21:101 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 2 33 gly nonfucosylated 409:422 arg1 IgG1-Fc 424:430 nonfucosylated IgG1-Fc 409:430 nonfucosylated IgG1-Fc 409:430 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 0 34 theme therapeutic 42:52 arg1 antibodies 54:63 therapeutic antibodies 42:63 therapeutic antibodies 42:63 Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. 22023369 1 35 theme improved 290:297 arg1 affinity 299:306 improved affinity 290:306 improved affinity for Fcγ receptor IIIa (FcγRIIIa) 290:339 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 5 36 theme structural 857:866 arg1 basis 868:872 a structural basis 855:872 a structural basis for improvement in ADCC of therapeutic antibodies by defucosylation 855:940 Our results offer a structural basis for improvement in ADCC of therapeutic antibodies by defucosylation. 22023369 4 37 theme steric 740:745 arg1 hindrance 747:755 steric hindrance 740:755 steric hindrance 740:755 However, fucosylation of the Fc N-glycans inhibits this interaction, because of steric hindrance, and furthermore, negatively affects the dynamics of the receptor binding site. 22023369 1 38 theme fucose 119:124 arg1 residue 126:132 the fucose residue 115:132 the fucose residue 115:132 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 3 39 theme nonfucosylated 608:621 arg1 Fc 623:624 nonfucosylated Fc 608:624 nonfucosylated Fc 608:624 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 22023369 3 40 theme sFcγRIIIa 568:576 arg1 N-glycans 555:563 the two N-glycans 547:563 the two N-glycans of sFcγRIIIa 547:576 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 22023369 1 41 theme residue 126:132 arg1 Removal 104:110 Removal 104:110 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) 104:195 Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) results in a dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) through improved affinity for Fcγ receptor IIIa (FcγRIIIa). 22023369 5 42 from improvement 878:888 arg1 ADCC 893:896 ADCC 893:896 ADCC of therapeutic antibodies by defucosylation 893:940 Our results offer a structural basis for improvement in ADCC of therapeutic antibodies by defucosylation. 22023369 2 43 theme FcγRIIIa 454:461 arg1 IgG1-Fc 424:430 nonfucosylated IgG1-Fc 409:430 nonfucosylated IgG1-Fc 409:430 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 2 43 theme FcγRIIIa 454:461 arg1 form 446:449 a soluble form 436:449 a soluble form of FcγRIIIa (sFcγRIIIa) 436:473 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 2 43 theme FcγRIIIa 454:461 arg1 sFcγRIIIa 464:472 sFcγRIIIa 464:472 sFcγRIIIa 464:472 Here, we present the 2.2-Å structure of the complex formed between nonfucosylated IgG1-Fc and a soluble form of FcγRIIIa (sFcγRIIIa) with two N-glycosylation sites. 22023369 5 44 theme antibodies 913:922 arg1 ADCC 893:896 ADCC 893:896 ADCC of therapeutic antibodies by defucosylation 893:940 Our results offer a structural basis for improvement in ADCC of therapeutic antibodies by defucosylation. 22023369 3 45 theme N-glycans 555:563 arg1 N-glycans 555:563 the two N-glycans 547:563 the two N-glycans of sFcγRIIIa 547:576 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 22023369 3 45 theme N-glycans 555:563 arg1 one 540:542 one 540:542 one 540:542 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 22023369 3 46 gly nonfucosylated 608:621 arg1 Fc 623:624 nonfucosylated Fc 608:624 nonfucosylated Fc 608:624 The crystal structure shows that one of the two N-glycans of sFcγRIIIa mediates the interaction with nonfucosylated Fc, thereby stabilizing the complex. 8636209 8 0 theme cells 968:972 arg1 treatment 953:961 treatment 953:961 treatment of T cells with tunicamycin 953:989 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 3 1 theme cell 412:415 arg1 expression 425:434 cell surface expression 412:434 cell surface expression of the complete TCR in human T cells 412:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 4 2 theme EC 510:511 arg1 domain 513:518 the EC domain 506:518 the EC domain of CD3 gamma 506:531 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 4 2 theme EC 510:511 arg1 gamma 527:531 CD3 gamma 523:531 CD3 gamma 523:531 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 10 3 theme TCR 1337:1339 arg1 expression 1319:1328 expression 1319:1328 expression 1319:1328 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 10 3 theme TCR 1337:1339 arg1 assembly 1306:1313 assembly 1306:1313 assembly 1306:1313 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 5 4 theme gamma 673:677 arg1 domain 659:664 the EC domain 652:664 the EC domain of CD3 gamma 652:677 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 5 4 theme gamma 673:677 arg1 gamma 673:677 CD3 gamma 669:677 CD3 gamma 669:677 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 1 5 theme CD3 134:136 arg1 epsilon 150:156 associated CD3 gamma delta epsilon 123:156 associated CD3 gamma delta epsilon 123:156 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 2 6 theme TCR 234:236 arg1 complex 238:244 the TCR complex 230:244 the TCR complex 230:244 The structural relationships between the subunits of the TCR complex are still not fully known. 8636209 5 7 theme EC 656:657 arg1 domain 659:664 the EC domain 652:664 the EC domain of CD3 gamma 652:677 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 5 7 theme EC 656:657 arg1 gamma 673:677 CD3 gamma 669:677 CD3 gamma 669:677 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 8 8 theme CD3 1032:1034 arg1 delta 1036:1040 CD3 delta 1032:1040 CD3 delta 1032:1040 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 7 9 theme gamma 885:889 arg1 glycosylation 864:876 glycosylation 864:876 glycosylation of CD3 gamma 864:889 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 3 10 theme human 459:463 arg1 cells 467:471 human T cells 459:471 human T cells 459:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 7 11 theme glycosylation 832:844 arg1 sites 846:850 N-linked glycosylation sites 823:850 N-linked glycosylation sites 823:850 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 6 12 theme TCR 759:761 arg1 assembly 763:770 TCR assembly 759:770 TCR assembly 759:770 Site-directed mutagenesis demonstrated that these sites play a crucial role in TCR assembly probably by binding to CD3 epsilon. 8636209 4 13 theme Ig 545:546 arg1 domain 548:553 an Ig domain 542:553 an Ig domain 542:553 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 3 14 theme TCR 452:454 arg1 expression 425:434 cell surface expression 412:434 cell surface expression of the complete TCR in human T cells 412:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 14 theme TCR 452:454 arg1 assembly 399:406 assembly 399:406 assembly 399:406 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 15 from expression 425:434 arg1 cells 467:471 human T cells 459:471 human T cells 459:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 4 16 theme gamma 527:531 arg1 domain 513:518 the EC domain 506:518 the EC domain of CD3 gamma 506:531 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 4 16 theme gamma 527:531 arg1 gamma 527:531 CD3 gamma 523:531 CD3 gamma 523:531 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 1 17 theme alpha 92:96 arg1 heterodimer 103:113 the Ti alpha beta heterodimer 85:113 the Ti alpha beta heterodimer 85:113 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 3 18 from assembly 399:406 arg1 cells 467:471 human T cells 459:471 human T cells 459:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 9 19 theme amino 1112:1116 arg1 acid 1118:1121 the acidic amino acid 1101:1121 the acidic amino acid 1101:1121 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 0 20 from Role 0:3 arg1 assembly 37:44 T cell receptor assembly 21:44 T cell receptor assembly 21:44 Role of CD3 gamma in T cell receptor assembly. 8636209 8 21 with treatment 953:961 arg1 tunicamycin 979:989 tunicamycin 979:989 tunicamycin 979:989 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 6 22 theme CD3 795:797 arg1 epsilon 799:805 CD3 epsilon 795:805 CD3 epsilon 795:805 Site-directed mutagenesis demonstrated that these sites play a crucial role in TCR assembly probably by binding to CD3 epsilon. 8636209 10 23 theme CY 1267:1268 arg1 domain 1270:1275 the entire CY domain 1256:1275 the entire CY domain of CD3 gamma 1256:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 10 23 theme CY 1267:1268 arg1 gamma 1284:1288 CD3 gamma 1280:1288 CD3 gamma 1280:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 3 24 theme transmembrane 335:347 arg1 domain 376:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain 311:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma 311:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 24 theme transmembrane 335:347 arg1 gamma 390:394 CD3 gamma 386:394 CD3 gamma 386:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 25 from role 303:306 arg1 expression 425:434 cell surface expression 412:434 cell surface expression of the complete TCR in human T cells 412:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 25 from role 303:306 arg1 assembly 399:406 assembly 399:406 assembly 399:406 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 1 26 theme T 51:51 arg1 receptor 58:65 The T cell receptor 47:65 The T cell receptor (TCR) 47:71 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 1 26 theme T 51:51 arg1 TCR 68:70 TCR 68:70 TCR 68:70 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 12 27 dep gamma 1524:1528 arg1 contrast 1508:1515 contrast 1508:1515 contrast 1508:1515 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 2 28 theme structural 181:190 arg1 relationships 192:204 The structural relationships 177:204 The structural relationships between the subunits of the TCR complex 177:244 The structural relationships between the subunits of the TCR complex are still not fully known. 8636209 10 29 theme gamma 1284:1288 arg1 domain 1270:1275 the entire CY domain 1256:1275 the entire CY domain of CD3 gamma 1256:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 10 29 theme gamma 1284:1288 arg1 gamma 1284:1288 CD3 gamma 1280:1288 CD3 gamma 1280:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 0 30 theme CD3 8:10 arg1 gamma 12:16 CD3 gamma 8:16 CD3 gamma 8:16 Role of CD3 gamma in T cell receptor assembly. 8636209 9 31 theme gamma 1147:1151 arg1 gamma 1147:1151 CD3 gamma 1143:1151 CD3 gamma 1143:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 9 31 theme gamma 1147:1151 arg1 domain 1133:1138 the TM domain 1126:1138 the TM domain of CD3 gamma 1126:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 6 32 theme Site-directed 680:692 arg1 mutagenesis 694:704 Site-directed mutagenesis 680:704 Site-directed mutagenesis 680:704 Site-directed mutagenesis demonstrated that these sites play a crucial role in TCR assembly probably by binding to CD3 epsilon. 8636209 11 33 theme gamma 1459:1463 arg1 EC 1435:1436 EC 1435:1436 EC 1435:1436 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 11 33 theme gamma 1459:1463 arg1 domain 1445:1450 TM domain 1442:1450 TM domain 1442:1450 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 11 33 theme gamma 1459:1463 arg1 gamma 1459:1463 CD3 gamma 1455:1463 CD3 gamma 1455:1463 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 9 34 theme Ti 1235:1236 arg1 beta 1238:1241 Ti beta 1235:1241 Ti beta 1235:1241 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 12 35 theme delta 1552:1556 arg1 glycosylation 1531:1543 glycosylation 1531:1543 glycosylation of CD3 delta 1531:1556 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 7 36 gly glycosylation 832:844 arg2 sites 846:850 N-linked glycosylation sites 823:850 N-linked glycosylation sites 823:850 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 5 37 theme interaction 616:626 arg1 sites 628:632 two potential interaction sites 602:632 two potential interaction sites 602:632 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 3 38 theme cytoplasmic 359:369 arg1 domain 376:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain 311:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma 311:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 38 theme cytoplasmic 359:369 arg1 gamma 390:394 CD3 gamma 386:394 CD3 gamma 386:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 1 39 theme zeta 162:165 arg1 chains 169:174 the associated CD3 gamma delta epsilon and zeta 2 chains 119:174 the associated CD3 gamma delta epsilon and zeta 2 chains 119:174 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 11 40 theme interaction 1399:1409 arg1 sites 1411:1415 specific TCR interaction sites 1386:1415 specific TCR interaction sites 1386:1415 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 3 41 theme CD3 386:388 arg1 gamma 390:394 CD3 gamma 386:394 CD3 gamma 386:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 9 42 theme TCR 1199:1201 arg1 assembly 1203:1210 TCR assembly 1199:1210 TCR assembly 1199:1210 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 5 43 theme potential 606:614 arg1 sites 628:632 two potential interaction sites 602:632 two potential interaction sites 602:632 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 6 44 theme crucial 743:749 arg1 role 751:754 a crucial role 741:754 a crucial role 741:754 Site-directed mutagenesis demonstrated that these sites play a crucial role in TCR assembly probably by binding to CD3 epsilon. 8636209 11 45 theme specific 1386:1393 arg1 sites 1411:1415 specific TCR interaction sites 1386:1415 specific TCR interaction sites 1386:1415 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 3 46 theme domain 376:381 arg1 role 303:306 the role 299:306 the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells 299:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 8 47 theme T 966:966 arg1 cells 968:972 T cells 966:972 T cells 966:972 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 12 48 theme CD3 1548:1550 arg1 delta 1552:1556 CD3 delta 1548:1556 CD3 delta 1548:1556 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 12 49 theme CD3 1520:1522 arg1 gamma 1524:1528 CD3 gamma 1520:1528 CD3 gamma 1520:1528 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 12 50 gly glycosylation 1531:1543 arg1 delta 1552:1556 CD3 delta 1548:1556 CD3 delta 1548:1556 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 12 50 gly glycosylation 1531:1543 arg1 TCR 1574:1576 TCR assembly 1574:1585 TCR assembly 1574:1585 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 11 51 theme TM 1442:1443 arg1 domain 1445:1450 TM domain 1442:1450 TM domain 1442:1450 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 11 51 theme TM 1442:1443 arg1 gamma 1459:1463 CD3 gamma 1455:1463 CD3 gamma 1455:1463 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 7 52 theme TCR 911:913 arg1 assembly 915:922 TCR assembly 911:922 TCR assembly 911:922 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 8 53 gly glycosylation 1015:1027 arg1 TCR 1058:1060 TCR assembly 1058:1069 TCR assembly 1058:1069 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 8 53 gly glycosylation 1015:1027 arg1 delta 1036:1040 CD3 delta 1032:1040 CD3 delta 1032:1040 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 5 54 theme CD3 669:671 arg1 gamma 673:677 CD3 gamma 669:677 CD3 gamma 669:677 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 0 55 theme cell 23:26 arg1 assembly 37:44 T cell receptor assembly 21:44 T cell receptor assembly 21:44 Role of CD3 gamma in T cell receptor assembly. 8636209 1 56 theme associated 123:132 arg1 epsilon 150:156 associated CD3 gamma delta epsilon 123:156 associated CD3 gamma delta epsilon 123:156 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 8 57 theme TCR 1058:1060 arg1 assembly 1062:1069 TCR assembly 1058:1069 TCR assembly 1058:1069 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 2 58 theme complex 238:244 arg1 subunits 218:225 the subunits 214:225 the subunits of the TCR complex 214:244 The structural relationships between the subunits of the TCR complex are still not fully known. 8636209 1 59 theme gamma 138:142 arg1 epsilon 150:156 associated CD3 gamma delta epsilon 123:156 associated CD3 gamma delta epsilon 123:156 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 8 60 theme N-linked 1006:1013 arg1 glycosylation 1015:1027 N-linked glycosylation 1006:1027 N-linked glycosylation of CD3 delta 1006:1040 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 4 61 theme computer 476:483 arg1 model 485:489 A computer model 474:489 A computer model 474:489 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 7 62 theme CD3 881:883 arg1 gamma 885:889 CD3 gamma 881:889 CD3 gamma 881:889 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 1 63 theme epsilon 150:156 arg1 chains 169:174 the associated CD3 gamma delta epsilon and zeta 2 chains 119:174 the associated CD3 gamma delta epsilon and zeta 2 chains 119:174 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 7 64 link N-linked 823:830 arg1 sites 846:850 N-linked glycosylation sites 823:850 N-linked glycosylation sites 823:850 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 3 65 theme complete 443:450 arg1 TCR 452:454 the complete TCR 439:454 the complete TCR 439:454 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 7 66 theme sites 846:850 arg1 Mutagenesis 808:818 Mutagenesis 808:818 Mutagenesis of N-linked glycosylation sites 808:850 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 8 67 theme delta 1036:1040 arg1 glycosylation 1015:1027 N-linked glycosylation 1006:1027 N-linked glycosylation of CD3 delta 1006:1040 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 7 68 gly glycosylation 864:876 arg1 TCR 911:913 TCR assembly 911:922 TCR assembly 911:922 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 7 68 gly glycosylation 864:876 arg1 gamma 885:889 CD3 gamma 881:889 CD3 gamma 881:889 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 9 69 theme Site-directed 1072:1084 arg1 mutagenesis 1086:1096 Site-directed mutagenesis 1072:1096 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma 1072:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 7 70 theme N-linked 823:830 arg1 sites 846:850 N-linked glycosylation sites 823:850 N-linked glycosylation sites 823:850 Mutagenesis of N-linked glycosylation sites showed that glycosylation of CD3 gamma is not required for TCR assembly and expression. 8636209 3 71 theme surface 417:423 arg1 expression 425:434 cell surface expression 412:434 cell surface expression of the complete TCR in human T cells 412:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 72 theme T 465:465 arg1 cells 467:471 human T cells 459:471 human T cells 459:471 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 1 73 theme Ti 89:90 arg1 heterodimer 103:113 the Ti alpha beta heterodimer 85:113 the Ti alpha beta heterodimer 85:113 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 4 74 theme CD3 523:525 arg1 gamma 527:531 CD3 gamma 523:531 CD3 gamma 523:531 A computer model indicated that the EC domain of CD3 gamma folds as an Ig domain. 8636209 5 75 theme alignment 583:591 arg1 studies 593:599 alignment studies 583:599 alignment studies 583:599 Based on this model and on alignment studies, two potential interaction sites were predicted in the EC domain of CD3 gamma. 8636209 9 76 theme acidic 1105:1110 arg1 acid 1118:1121 the acidic amino acid 1101:1121 the acidic amino acid 1101:1121 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 1 77 theme beta 98:101 arg1 heterodimer 103:113 the Ti alpha beta heterodimer 85:113 the Ti alpha beta heterodimer 85:113 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 9 78 theme acid 1118:1121 arg1 mutagenesis 1086:1096 Site-directed mutagenesis 1072:1096 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma 1072:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 8 79 link N-linked 1006:1013 arg1 glycosylation 1015:1027 N-linked glycosylation 1006:1027 N-linked glycosylation of CD3 delta 1006:1040 In contrast, treatment of T cells with tunicamycin suggested that N-linked glycosylation of CD3 delta is required for TCR assembly. 8636209 3 80 dep cytoplasmic 359:369 arg1 CY 372:373 CY 372:373 CY 372:373 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 1 81 theme delta 144:148 arg1 epsilon 150:156 associated CD3 gamma delta epsilon 123:156 associated CD3 gamma delta epsilon 123:156 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 3 82 dep extracellular 315:327 arg1 EC 330:331 EC 330:331 EC 330:331 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 1 83 theme cell 53:56 arg1 receptor 58:65 The T cell receptor 47:65 The T cell receptor (TCR) 47:71 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 1 83 theme cell 53:56 arg1 TCR 68:70 TCR 68:70 TCR 68:70 The T cell receptor (TCR) consists of the Ti alpha beta heterodimer and the associated CD3 gamma delta epsilon and zeta 2 chains. 8636209 0 84 theme gamma 12:16 arg1 Role 0:3 Role 0:3 Role of CD3 gamma in T cell receptor assembly. 0:45 Role of CD3 gamma in T cell receptor assembly. 8636209 9 85 theme CD3 1143:1145 arg1 gamma 1147:1151 CD3 gamma 1143:1151 CD3 gamma 1143:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 10 86 theme CD3 1280:1282 arg1 gamma 1284:1288 CD3 gamma 1280:1288 CD3 gamma 1280:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 11 87 theme CD3 1455:1457 arg1 gamma 1459:1463 CD3 gamma 1455:1463 CD3 gamma 1455:1463 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 0 88 theme T 21:21 arg1 assembly 37:44 T cell receptor assembly 21:44 T cell receptor assembly 21:44 Role of CD3 gamma in T cell receptor assembly. 8636209 10 89 theme domain 1270:1275 arg1 Deletion 1244:1251 Deletion 1244:1251 Deletion of the entire CY domain of CD3 gamma 1244:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 0 90 theme receptor 28:35 arg1 assembly 37:44 T cell receptor assembly 21:44 T cell receptor assembly 21:44 Role of CD3 gamma in T cell receptor assembly. 8636209 11 91 dep EC 1435:1436 arg1 the 1431:1433 the 1431:1433 the 1431:1433 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 12 92 theme TCR 1574:1576 arg1 assembly 1578:1585 TCR assembly 1574:1585 TCR assembly 1574:1585 Furthermore, the study indicated that, in contrast to CD3 gamma, glycosylation of CD3 delta is required for TCR assembly and expression. 8636209 3 93 dep transmembrane 335:347 arg1 TM 350:351 TM 350:351 TM 350:351 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 9 94 theme TM 1130:1131 arg1 gamma 1147:1151 CD3 gamma 1143:1151 CD3 gamma 1143:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 9 94 theme TM 1130:1131 arg1 domain 1133:1138 the TM domain 1126:1138 the TM domain of CD3 gamma 1126:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 3 95 theme extracellular 315:327 arg1 domain 376:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain 311:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma 311:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 95 theme extracellular 315:327 arg1 gamma 390:394 CD3 gamma 386:394 CD3 gamma 386:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 10 96 theme entire 1260:1265 arg1 domain 1270:1275 the entire CY domain 1256:1275 the entire CY domain of CD3 gamma 1256:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 10 96 theme entire 1260:1265 arg1 gamma 1284:1288 CD3 gamma 1280:1288 CD3 gamma 1280:1288 Deletion of the entire CY domain of CD3 gamma did not prevent assembly and expression of the TCR. 8636209 11 97 theme TCR 1395:1397 arg1 sites 1411:1415 specific TCR interaction sites 1386:1415 specific TCR interaction sites 1386:1415 In conclusion, this study demonstrated that specific TCR interaction sites exist in both the EC and TM domain of CD3 gamma. 8636209 9 98 from mutagenesis 1086:1096 arg1 gamma 1147:1151 CD3 gamma 1143:1151 CD3 gamma 1143:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 9 98 from mutagenesis 1086:1096 arg1 domain 1133:1138 the TM domain 1126:1138 the TM domain of CD3 gamma 1126:1151 Site-directed mutagenesis of the acidic amino acid in the TM domain of CD3 gamma demonstrated that this residue is involved in TCR assembly probably by binding to Ti beta. 8636209 3 99 theme gamma 390:394 arg1 domain 376:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain 311:381 the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma 311:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 8636209 3 99 theme gamma 390:394 arg1 gamma 390:394 CD3 gamma 386:394 CD3 gamma 386:394 In this study we examined the role of the extracellular (EC), transmembrane (TM), and cytoplasmic (CY) domain of CD3 gamma in assembly and cell surface expression of the complete TCR in human T cells. 10091666 4 0 theme glycosyl 826:833 arg1 group 835:839 the glycosyl group 822:839 the glycosyl group 822:839 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 5 1 gly deglycosylated 998:1011 arg1 proteins 1023:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 0 2 theme alpha-amylase 66:78 arg1 mutagenesis 9:19 mutagenesis 9:19 mutagenesis 9:19 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 0 2 theme alpha-amylase 66:78 arg1 Cloning 0:6 Cloning 0:6 Cloning 0:6 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 0 2 theme alpha-amylase 66:78 arg1 analysis 37:44 structural analysis 26:44 structural analysis 26:44 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 4 3 theme crystal 753:759 arg1 structure 761:769 the crystal structure 749:769 the crystal structure 749:769 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 0 4 theme Pichia 93:98 arg1 pastoris 100:107 Pichia pastoris 93:107 Pichia pastoris 93:107 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 3 5 theme single 670:675 arg1 GlcNAc 699:704 GlcNAc 699:704 GlcNAc 699:704 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 3 5 theme single 670:675 arg1 glucosamine 686:696 a single N-acetyl glucosamine 668:696 a single N-acetyl glucosamine (GlcNAc) 668:705 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 4 6 theme A 794:794 arg1 resolution 776:785 a resolution 774:785 a resolution of 2.0 A 774:794 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 6 7 theme crucial 1222:1228 arg1 role 1230:1233 a crucial role 1220:1233 a crucial role 1220:1233 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 6 7 theme crucial 1222:1228 arg1 nucleophile 1298:1308 the catalytic nucleophile 1284:1308 the catalytic nucleophile 1284:1308 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 5 8 theme k 1056:1056 arg1 values 1066:1071 the k(cat)/Km values 1052:1071 the k(cat)/Km values for D197A and D197N 1052:1091 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 8 theme k 1056:1056 arg1 times 1110:1114 10(6)-10(7) times 1098:1114 10(6)-10(7) times lower than the wild-type enzyme 1098:1146 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 4 9 theme group 835:839 arg1 location 810:817 the location 806:817 the location of the glycosyl group 806:839 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 6 10 theme k 1163:1163 arg1 values 1173:1178 The decreased k(cat)/Km values 1149:1178 The decreased k(cat)/Km values for the mutants 1149:1194 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 6 11 theme decreased 1153:1161 arg1 values 1173:1178 The decreased k(cat)/Km values 1149:1178 The decreased k(cat)/Km values for the mutants 1149:1194 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 2 12 theme mass 372:375 arg1 spectrometry 377:388 electrospray ionization mass spectrometry 348:388 electrospray ionization mass spectrometry (ESI-MS) 348:397 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 12 theme mass 372:375 arg1 ESI-MS 391:396 ESI-MS 391:396 ESI-MS 391:396 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 13 theme peptide 471:477 arg1 mapping 479:485 peptide mapping 471:485 peptide mapping 471:485 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 3 14 with product 655:661 arg1 GlcNAc 699:704 GlcNAc 699:704 GlcNAc 699:704 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 3 14 with product 655:661 arg1 glucosamine 686:696 a single N-acetyl glucosamine 668:696 a single N-acetyl glucosamine (GlcNAc) 668:705 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 4 15 theme native 936:941 arg1 conformation 916:927 essentially the same conformation 895:927 essentially the same conformation 895:927 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 4 15 theme native 936:941 arg1 enzyme 943:948 the native enzyme 932:948 the native enzyme 932:948 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 6 16 theme hydrolytic 1242:1251 arg1 activity 1253:1260 the hydrolytic activity 1238:1260 the hydrolytic activity of HPA 1238:1267 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 5 17 theme kinetic 955:961 arg1 same 1041:1044 same 1041:1044 same 1041:1044 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 17 theme kinetic 955:961 arg1 parameters 963:972 The kinetic parameters 951:972 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins 951:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 3 18 theme proteins 534:541 arg1 Treatment 515:523 Treatment 515:523 Treatment of these proteins with endoglycosidase F 515:564 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 1 19 theme methylotrophic 168:181 arg1 pastoris 196:203 the methylotrophic yeast Pichia pastoris 164:203 the methylotrophic yeast Pichia pastoris 164:203 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 6 20 theme catalytic 1288:1296 arg1 role 1230:1233 a crucial role 1220:1233 a crucial role 1220:1233 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 6 20 theme catalytic 1288:1296 arg1 nucleophile 1298:1308 the catalytic nucleophile 1284:1308 the catalytic nucleophile 1284:1308 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 3 21 theme protein 647:653 arg1 product 655:661 a protein product 645:661 a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized 645:734 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 2 22 with conjunction 490:500 arg1 ESI-MS 507:512 ESI-MS 507:512 ESI-MS 507:512 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 1 23 theme yeast 183:187 arg1 pastoris 196:203 the methylotrophic yeast Pichia pastoris 164:203 the methylotrophic yeast Pichia pastoris 164:203 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 24 theme conserved 255:263 arg1 acid 288:291 a completely conserved active site carboxylic acid 242:291 a completely conserved active site carboxylic acid 242:291 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 4 25 theme recombinant 871:881 arg1 protein 883:889 the recombinant protein 867:889 the recombinant protein 867:889 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 4 26 contain had 891:893 arg2 enzyme 943:948 the native enzyme 932:948 the native enzyme 932:948 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 4 26 contain had 891:893 arg2 conformation 916:927 essentially the same conformation 895:927 essentially the same conformation 895:927 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 4 26 contain had 891:893 arg1 protein 883:889 the recombinant protein 867:889 the recombinant protein 867:889 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 1 27 theme Pichia 189:194 arg1 pastoris 196:203 the methylotrophic yeast Pichia pastoris 164:203 the methylotrophic yeast Pichia pastoris 164:203 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 5 28 gly glycosylated 981:992 arg1 proteins 1023:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 1 29 theme active 265:270 arg1 acid 288:291 a completely conserved active site carboxylic acid 242:291 a completely conserved active site carboxylic acid 242:291 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 3 30 theme N-linked 608:615 arg1 oligosaccharide 617:631 a single N-linked oligosaccharide 599:631 a single N-linked oligosaccharide 599:631 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 0 31 theme structural 26:35 arg1 analysis 37:44 structural analysis 26:44 structural analysis 26:44 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 1 32 theme site 272:275 arg1 acid 288:291 a completely conserved active site carboxylic acid 242:291 a completely conserved active site carboxylic acid 242:291 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 2 33 theme ionization 361:370 arg1 spectrometry 377:388 electrospray ionization mass spectrometry 348:388 electrospray ionization mass spectrometry (ESI-MS) 348:397 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 33 theme ionization 361:370 arg1 ESI-MS 391:396 ESI-MS 391:396 ESI-MS 391:396 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 1 34 theme carboxylic 277:286 arg1 acid 288:291 a completely conserved active site carboxylic acid 242:291 a completely conserved active site carboxylic acid 242:291 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 35 dep mutants 213:219 arg1 D197A 222:226 D197A 222:226 D197A 222:226 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 35 dep mutants 213:219 arg1 mutants 213:219 two mutants 209:219 two mutants (D197A and D197N) of a completely conserved active site carboxylic acid 209:291 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 35 dep mutants 213:219 arg1 D197N 232:236 D197N 232:236 D197N 232:236 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 5 36 theme -10 1103:1105 arg1 values 1066:1071 the k(cat)/Km values 1052:1071 the k(cat)/Km values for D197A and D197N 1052:1091 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 36 theme -10 1103:1105 arg1 times 1110:1114 10(6)-10(7) times 1098:1114 10(6)-10(7) times lower than the wild-type enzyme 1098:1146 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 4 37 theme same 911:914 arg1 conformation 916:927 essentially the same conformation 895:927 essentially the same conformation 895:927 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 4 37 theme same 911:914 arg1 enzyme 943:948 the native enzyme 932:948 the native enzyme 932:948 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 2 38 theme attachment 434:443 arg1 Asn461 461:466 Asn461 461:466 Asn461 461:466 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 38 theme attachment 434:443 arg1 attachment 434:443 attachment 434:443 attachment 434:443 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 38 theme attachment 434:443 arg1 site 426:429 the site 422:429 the site of attachment 422:443 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 1 39 theme acid 288:291 arg1 D197A 222:226 D197A 222:226 D197A 222:226 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 39 theme acid 288:291 arg1 mutants 213:219 two mutants 209:219 two mutants (D197A and D197N) of a completely conserved active site carboxylic acid 209:291 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 39 theme acid 288:291 arg1 D197N 232:236 D197N 232:236 D197N 232:236 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 2 40 from Asn461 461:466 arg1 conjunction 490:500 conjunction 490:500 conjunction with ESI-MS 490:512 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 3 41 theme endoglycosidase 548:562 arg1 F 564:564 endoglycosidase F 548:564 endoglycosidase F 548:564 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 5 42 theme glycosylated 981:992 arg1 proteins 1023:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 1 43 theme Human 110:114 arg1 alpha-amylase 127:139 Human pancreatic alpha-amylase 110:139 Human pancreatic alpha-amylase (HPA) 110:145 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 43 theme Human 110:114 arg1 HPA 142:144 HPA 142:144 HPA 142:144 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 3 44 with Treatment 515:523 arg1 F 564:564 endoglycosidase F 548:564 endoglycosidase F 548:564 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 6 45 theme HPA 1265:1267 arg1 activity 1253:1260 the hydrolytic activity 1238:1260 the hydrolytic activity of HPA 1238:1267 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 5 46 theme /Km 1062:1064 arg1 values 1066:1071 the k(cat)/Km values 1052:1071 the k(cat)/Km values for D197A and D197N 1052:1091 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 46 theme /Km 1062:1064 arg1 times 1110:1114 10(6)-10(7) times 1098:1114 10(6)-10(7) times lower than the wild-type enzyme 1098:1146 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 47 theme deglycosylated 998:1011 arg1 proteins 1023:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 1 48 theme pancreatic 116:125 arg1 alpha-amylase 127:139 Human pancreatic alpha-amylase 110:139 Human pancreatic alpha-amylase (HPA) 110:145 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 1 48 theme pancreatic 116:125 arg1 HPA 142:144 HPA 142:144 HPA 142:144 Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. 10091666 5 49 theme wild-type 1013:1021 arg1 proteins 1023:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 the glycosylated and deglycosylated wild-type proteins 977:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 4 50 theme structure 761:769 arg1 Solution 737:744 Solution 737:744 Solution of the crystal structure to a resolution of 2.0 A 737:794 Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. 10091666 3 51 contain contained 589:597 arg2 oligosaccharide 617:631 a single N-linked oligosaccharide 599:631 a single N-linked oligosaccharide 599:631 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 3 51 contain contained 589:597 arg1 they 584:587 they 584:587 they 584:587 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 6 52 theme /Km 1169:1171 arg1 values 1173:1178 The decreased k(cat)/Km values 1149:1178 The decreased k(cat)/Km values for the mutants 1149:1194 The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile. 10091666 5 53 theme proteins 1023:1030 arg1 same 1041:1044 same 1041:1044 same 1041:1044 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 53 theme proteins 1023:1030 arg1 parameters 963:972 The kinetic parameters 951:972 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins 951:1030 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 3 54 theme single 601:606 arg1 oligosaccharide 617:631 a single N-linked oligosaccharide 599:631 a single N-linked oligosaccharide 599:631 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 0 55 theme pancreatic 55:64 arg1 alpha-amylase 66:78 human pancreatic alpha-amylase 49:78 human pancreatic alpha-amylase 49:78 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 2 56 theme recombinant 313:323 arg1 proteins 325:332 All recombinant proteins 309:332 All recombinant proteins 309:332 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 57 gly glycosylated 405:416 arg1 proteins 325:332 All recombinant proteins 309:332 All recombinant proteins 309:332 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 58 theme electrospray 348:359 arg1 spectrometry 377:388 electrospray ionization mass spectrometry 348:388 electrospray ionization mass spectrometry (ESI-MS) 348:397 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 2 58 theme electrospray 348:359 arg1 ESI-MS 391:396 ESI-MS 391:396 ESI-MS 391:396 All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. 10091666 0 59 theme human 49:53 arg1 alpha-amylase 66:78 human pancreatic alpha-amylase 49:78 human pancreatic alpha-amylase 49:78 Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. 10091666 3 60 theme N-acetyl 677:684 arg1 GlcNAc 699:704 GlcNAc 699:704 GlcNAc 699:704 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 3 60 theme N-acetyl 677:684 arg1 glucosamine 686:696 a single N-acetyl glucosamine 668:696 a single N-acetyl glucosamine (GlcNAc) 668:705 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 5 61 theme lower 1116:1120 arg1 values 1066:1071 the k(cat)/Km values 1052:1071 the k(cat)/Km values for D197A and D197N 1052:1091 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 5 61 theme lower 1116:1120 arg1 times 1110:1114 10(6)-10(7) times 1098:1114 10(6)-10(7) times lower than the wild-type enzyme 1098:1146 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 10091666 3 62 link N-linked 608:615 arg1 oligosaccharide 617:631 a single N-linked oligosaccharide 599:631 a single N-linked oligosaccharide 599:631 Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. 10091666 5 63 theme wild-type 1131:1139 arg1 enzyme 1141:1146 the wild-type enzyme 1127:1146 the wild-type enzyme 1127:1146 The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. 16467855 2 0 theme sclerosis 509:517 arg1 models 471:476 preclinical models 459:476 preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain 459:539 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 2 1 from neuroprotection 440:454 arg1 models 471:476 preclinical models 459:476 preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain 459:539 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 3 2 theme hydrolysis 708:717 arg1 reaction 719:726 the enzyme's hydrolysis reaction 695:726 the enzyme's hydrolysis reaction 695:726 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 2 3 theme GCPII 367:371 arg1 inhibitors 373:382 Potent and selective GCPII inhibitors 346:382 Potent and selective GCPII inhibitors 346:382 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 0 4 theme neuronal 61:68 arg1 damage 70:75 neuronal damage 61:75 neuronal damage 61:75 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 4 5 theme apical 825:830 arg1 domains 801:807 three domains 795:807 three domains: protease-like, apical, and C-terminal 795:846 GCPII folds into three domains: protease-like, apical, and C-terminal. 16467855 2 6 theme selective 357:365 arg1 inhibitors 373:382 Potent and selective GCPII inhibitors 346:382 Potent and selective GCPII inhibitors 346:382 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 0 7 theme prostate 81:88 arg1 cancer 90:95 prostate cancer 81:95 prostate cancer 81:95 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 4 8 theme C-terminal 837:846 arg1 domains 801:807 three domains 795:807 three domains: protease-like, apical, and C-terminal 795:846 GCPII folds into three domains: protease-like, apical, and C-terminal. 16467855 7 9 theme prostate 1289:1296 arg1 cancer 1298:1303 prostate cancer 1289:1303 prostate cancer 1289:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 7 10 theme essential 1183:1191 arg1 information 1193:1203 essential information 1183:1203 essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer 1183:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 3 11 theme reaction 719:726 arg1 glutamate 669:677 glutamate 669:677 glutamate 669:677 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 3 11 theme reaction 719:726 arg1 product 684:690 the product 680:690 the product of the enzyme's hydrolysis reaction 680:726 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 7 12 theme GCPII 1223:1227 arg1 inhibitors 1229:1238 GCPII inhibitors 1223:1238 GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer 1223:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 3 13 theme crystal 558:564 arg1 structures 566:575 crystal structures 558:575 crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively 558:775 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 2 14 theme pain 536:539 arg1 models 471:476 preclinical models 459:476 preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain 459:539 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 6 15 theme GCPII 1052:1056 arg1 formation 1039:1047 homodimer formation 1029:1047 homodimer formation of GCPII 1029:1056 One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. 16467855 2 16 theme neuropathic 524:534 arg1 pain 536:539 neuropathic pain 524:539 neuropathic pain 524:539 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 2 17 theme lateral 501:507 arg1 sclerosis 509:517 amyotrophic lateral sclerosis 489:517 amyotrophic lateral sclerosis 489:517 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 7 18 theme induced-fit 1117:1127 arg1 mode 1147:1150 an induced-fit substrate-binding mode 1114:1150 an induced-fit substrate-binding mode of this key enzyme 1114:1169 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 5 19 theme C-terminal 931:940 arg1 recognition 952:962 C-terminal glutamate recognition 931:962 C-terminal glutamate recognition 931:962 All three participate in substrate binding, with two of them directly involved in C-terminal glutamate recognition. 16467855 1 20 theme zinc 156:159 arg1 metalloenzyme 161:173 a zinc metalloenzyme 154:173 a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter) 154:343 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 1 20 theme zinc 156:159 arg1 GCPII 144:148 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 1 21 dep itself 318:323 arg1 neurotransmitter 327:342 a neurotransmitter 325:342 a neurotransmitter 325:342 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 7 22 theme cancer 1298:1303 arg1 treatment 1254:1262 the treatment 1250:1262 the treatment of neuronal diseases and prostate cancer 1250:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 2 23 theme preclinical 459:469 arg1 models 471:476 preclinical models 459:476 preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain 459:539 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 4 24 theme protease-like 810:822 arg1 domains 801:807 three domains 795:807 three domains: protease-like, apical, and C-terminal 795:846 GCPII folds into three domains: protease-like, apical, and C-terminal. 16467855 7 25 from treatment 1254:1262 arg1 useful 1240:1245 useful 1240:1245 useful 1240:1245 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 3 26 theme extracellular 584:596 arg1 part 598:601 the extracellular part 580:601 the extracellular part of GCPII 580:610 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 0 27 theme carboxypeptidase 23:38 arg1 Structure 0:8 Structure 0:8 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer 0:95 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 7 28 from useful 1240:1245 arg1 treatment 1254:1262 the treatment 1250:1262 the treatment of neuronal diseases and prostate cancer 1250:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 0 29 theme glutamate 13:21 arg1 target 51:56 a drug target 44:56 a drug target in neuronal damage and prostate cancer 44:95 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 0 29 theme glutamate 13:21 arg1 carboxypeptidase 23:38 glutamate carboxypeptidase II 13:41 glutamate carboxypeptidase II 13:41 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 5 30 theme them 905:908 arg1 them 905:908 them 905:908 them 905:908 All three participate in substrate binding, with two of them directly involved in C-terminal glutamate recognition. 16467855 5 30 theme them 905:908 arg1 two 898:900 two 898:900 two 898:900 All three participate in substrate binding, with two of them directly involved in C-terminal glutamate recognition. 16467855 6 31 theme enzyme 1005:1010 arg1 moieties 989:996 the carbohydrate moieties 972:996 the carbohydrate moieties of the enzyme 972:1010 One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. 16467855 2 32 theme amyotrophic 489:499 arg1 sclerosis 509:517 amyotrophic lateral sclerosis 489:517 amyotrophic lateral sclerosis 489:517 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 2 33 theme Potent 346:351 arg1 inhibitors 373:382 Potent and selective GCPII inhibitors 346:382 Potent and selective GCPII inhibitors 346:382 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 7 34 theme key 1160:1162 arg1 enzyme 1164:1169 this key enzyme 1155:1169 this key enzyme 1155:1169 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 3 35 theme GCPII 606:610 arg1 part 598:601 the extracellular part 580:601 the extracellular part of GCPII 580:610 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 7 36 theme neuronal 1267:1274 arg1 diseases 1276:1283 neuronal diseases 1267:1283 neuronal diseases 1267:1283 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 7 37 theme substrate-binding 1129:1145 arg1 mode 1147:1150 an induced-fit substrate-binding mode 1114:1150 an induced-fit substrate-binding mode of this key enzyme 1114:1169 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 5 38 theme substrate 874:882 arg1 binding 884:890 substrate binding 874:890 substrate binding 874:890 All three participate in substrate binding, with two of them directly involved in C-terminal glutamate recognition. 16467855 2 39 theme brain 412:416 arg1 glutamate 418:426 brain glutamate 412:426 brain glutamate 412:426 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 0 40 theme drug 46:49 arg1 target 51:56 a drug target 44:56 a drug target in neuronal damage and prostate cancer 44:95 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 0 40 theme drug 46:49 arg1 carboxypeptidase 23:38 glutamate carboxypeptidase II 13:41 glutamate carboxypeptidase II 13:41 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 3 41 from resolution 752:761 arg1 glutamate 669:677 glutamate 669:677 glutamate 669:677 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 3 41 from resolution 752:761 arg1 product 684:690 the product 680:690 the product of the enzyme's hydrolysis reaction 680:726 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 0 42 from target 51:56 arg1 damage 70:75 neuronal damage 61:75 neuronal damage 61:75 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 0 42 from target 51:56 arg1 cancer 90:95 prostate cancer 81:95 prostate cancer 81:95 Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. 16467855 3 43 theme part 598:601 arg1 structures 566:575 crystal structures 558:575 crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively 558:775 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 1 44 theme Membrane-bound 98:111 arg1 metalloenzyme 161:173 a zinc metalloenzyme 154:173 a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter) 154:343 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 1 44 theme Membrane-bound 98:111 arg1 GCPII 144:148 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 3 45 theme weak 644:647 arg1 inhibitors 649:658 both potent and weak inhibitors 628:658 both potent and weak inhibitors 628:658 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 1 46 theme glutamate 113:121 arg1 metalloenzyme 161:173 a zinc metalloenzyme 154:173 a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter) 154:343 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 1 46 theme glutamate 113:121 arg1 GCPII 144:148 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 3 47 from structures 566:575 arg1 complex 615:621 complex 615:621 complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively 615:775 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 5 48 theme glutamate 942:950 arg1 recognition 952:962 C-terminal glutamate recognition 931:962 C-terminal glutamate recognition 931:962 All three participate in substrate binding, with two of them directly involved in C-terminal glutamate recognition. 16467855 6 49 theme carbohydrate 976:987 arg1 moieties 989:996 the carbohydrate moieties 972:996 the carbohydrate moieties of the enzyme 972:1010 One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. 16467855 1 50 theme carboxypeptidase 123:138 arg1 metalloenzyme 161:173 a zinc metalloenzyme 154:173 a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter) 154:343 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 1 50 theme carboxypeptidase 123:138 arg1 GCPII 144:148 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) 98:149 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 6 51 theme moieties 989:996 arg1 One 965:967 One 965:967 One 965:967 One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. 16467855 6 51 theme moieties 989:996 arg1 moieties 989:996 the carbohydrate moieties 972:996 the carbohydrate moieties of the enzyme 972:1010 One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. 16467855 3 52 theme potent 633:638 arg1 inhibitors 649:658 both potent and weak inhibitors 628:658 both potent and weak inhibitors 628:658 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 7 53 theme three-dimensional 1063:1079 arg1 structures 1081:1090 The three-dimensional structures 1059:1090 The three-dimensional structures presented here 1059:1105 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 1 54 theme neurotransmitter 212:227 arg1 N-acetyl-L-aspartyl-L-glutamate 229:259 the neurotransmitter N-acetyl-L-aspartyl-L-glutamate 208:259 the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) 208:266 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 1 54 theme neurotransmitter 212:227 arg1 NAAG 262:265 NAAG 262:265 NAAG 262:265 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 3 55 with complex 615:621 arg1 inhibitors 649:658 both potent and weak inhibitors 628:658 both potent and weak inhibitors 628:658 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 3 55 with complex 615:621 arg1 glutamate 669:677 glutamate 669:677 glutamate 669:677 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 3 55 with complex 615:621 arg1 product 684:690 the product 680:690 the product of the enzyme's hydrolysis reaction 680:726 Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. 16467855 7 56 theme diseases 1276:1283 arg1 treatment 1254:1262 the treatment 1250:1262 the treatment of neuronal diseases and prostate cancer 1250:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 7 57 theme inhibitors 1229:1238 arg1 design 1213:1218 the design 1209:1218 the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer 1209:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 6 58 theme homodimer 1029:1037 arg1 formation 1039:1047 homodimer formation 1029:1047 homodimer formation of GCPII 1029:1056 One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. 16467855 1 59 theme N-acetyl-L-aspartyl-L-glutamate 229:259 arg1 hydrolysis 194:203 the hydrolysis 190:203 the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter) 190:343 Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). 16467855 2 60 theme stroke 481:486 arg1 models 471:476 preclinical models 459:476 preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain 459:539 Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. 16467855 7 61 theme enzyme 1164:1169 arg1 mode 1147:1150 an induced-fit substrate-binding mode 1114:1150 an induced-fit substrate-binding mode of this key enzyme 1114:1169 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 16467855 7 62 theme useful 1240:1245 arg1 inhibitors 1229:1238 GCPII inhibitors 1223:1238 GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer 1223:1303 The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. 10419520 7 0 theme face 1078:1081 arg1 2 1051:1051 2 1051:1051 2 1051:1051 Both the NH(2) and COOH termini of S2P face the cytosol. 10419520 7 0 theme face 1078:1081 arg1 NH 1048:1049 the NH 1044:1049 the NH(2) 1044:1052 Both the NH(2) and COOH termini of S2P face the cytosol. 10419520 7 0 theme face 1078:1081 arg1 termini 1063:1069 COOH termini 1058:1069 COOH termini of S2P face the cytosol 1058:1093 Both the NH(2) and COOH termini of S2P face the cytosol. 10419520 6 1 from topology 1006:1013 arg1 membranes 1028:1036 ER membranes 1025:1036 ER membranes 1025:1036 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 1 2 theme regulatory 234:243 arg1 SREBPs 271:276 SREBPs 271:276 SREBPs 271:276 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 2 theme regulatory 234:243 arg1 proteins 261:268 sterol regulatory element-binding proteins 227:268 sterol regulatory element-binding proteins (SREBPs) 227:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 10 3 theme HEIGH 1290:1294 arg1 sequence 1296:1303 The HEIGH sequence 1286:1303 The HEIGH sequence 1286:1303 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 3 4 from membrane 474:481 arg1 fashion 496:502 a hairpin fashion 486:502 a hairpin fashion 486:502 The SREBPs are oriented in the membrane in a hairpin fashion, with the NH(2)- and COOH-terminal domains facing the cytosol and a single hydrophilic loop projecting into the lumen. 10419520 1 5 theme two-step 159:166 arg1 process 180:186 a two-step proteolytic process 157:186 a two-step proteolytic process 157:186 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 6 theme element-binding 245:259 arg1 SREBPs 271:276 SREBPs 271:276 SREBPs 271:276 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 6 theme element-binding 245:259 arg1 proteins 261:268 sterol regulatory element-binding proteins 227:268 sterol regulatory element-binding proteins (SREBPs) 227:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 0 7 theme regulatory 84:93 arg1 proteins 111:118 sterol regulatory element-binding proteins 77:118 sterol regulatory element-binding proteins 77:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 4 8 theme first 800:804 arg1 domain 820:825 the first transmembrane domain 796:825 the first transmembrane domain 796:825 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 5 9 theme zinc 875:878 arg1 metalloprotease 880:894 a putative zinc metalloprotease 864:894 a putative zinc metalloprotease 864:894 A membrane protein, designated S2P, a putative zinc metalloprotease, is required for this cleavage. 10419520 5 9 theme zinc 875:878 arg1 S2P 859:861 S2P 859:861 S2P 859:861 A membrane protein, designated S2P, a putative zinc metalloprotease, is required for this cleavage. 10419520 5 10 theme putative 866:873 arg1 metalloprotease 880:894 a putative zinc metalloprotease 864:894 a putative zinc metalloprotease 864:894 A membrane protein, designated S2P, a putative zinc metalloprotease, is required for this cleavage. 10419520 5 10 theme putative 866:873 arg1 S2P 859:861 S2P 859:861 S2P 859:861 A membrane protein, designated S2P, a putative zinc metalloprotease, is required for this cleavage. 10419520 1 11 theme proteins 261:268 arg1 proteins 261:268 sterol regulatory element-binding proteins 227:268 sterol regulatory element-binding proteins (SREBPs) 227:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 11 theme proteins 261:268 arg1 domains 216:222 the NH(2)-terminal domains 197:222 the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) 197:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 0 12 theme sterol 77:82 arg1 proteins 111:118 sterol regulatory element-binding proteins 77:118 sterol regulatory element-binding proteins 77:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 13 13 theme transmembrane 1812:1824 arg1 helix 1826:1830 the first transmembrane helix 1802:1830 the first transmembrane helix of SREBPs 1802:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 11 14 theme located 1437:1443 arg1 acid 1427:1430 Aspartic acid 467 1418:1434 Aspartic acid 467 1418:1434 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 13 15 theme sequences 1663:1671 arg1 hydrophobicity 1639:1652 The hydrophobicity 1635:1652 The hydrophobicity of these sequences 1635:1671 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 6 16 theme S2P 1018:1020 arg1 topology 1006:1013 the topology 1002:1013 the topology of S2P in ER membranes 1002:1036 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 0 17 theme proteins 111:118 arg1 cleavage 65:72 intramembranous cleavage 49:72 intramembranous cleavage of sterol regulatory element-binding proteins 49:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 11 18 theme HEIGH 1486:1490 arg1 sequence 1492:1499 the HEIGH sequence 1482:1499 the HEIGH sequence 1482:1499 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 1 19 theme proteolytic 168:178 arg1 process 180:186 a two-step proteolytic process 157:186 a two-step proteolytic process 157:186 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 0 20 theme element-binding 95:109 arg1 proteins 111:118 sterol regulatory element-binding proteins 77:118 sterol regulatory element-binding proteins 77:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 4 21 from Site-2 770:775 arg1 cleavage 758:765 cleavage 758:765 cleavage at Site-2, which lies within the first transmembrane domain 758:825 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 0 22 theme Membrane 0:7 arg1 protein 28:34 a protein 26:34 a protein required for intramembranous cleavage of sterol regulatory element-binding proteins 26:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 0 22 theme Membrane 0:7 arg1 topology 9:16 Membrane topology 0:16 Membrane topology of S2P 0:23 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 13 23 from bond 1794:1797 arg1 helix 1826:1830 the first transmembrane helix 1802:1830 the first transmembrane helix of SREBPs 1802:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 7 24 theme COOH 1058:1061 arg1 termini 1063:1069 COOH termini 1058:1069 COOH termini of S2P face the cytosol 1058:1093 Both the NH(2) and COOH termini of S2P face the cytosol. 10419520 2 25 theme acid 424:427 arg1 biosynthesis 429:440 fatty acid biosynthesis 418:440 fatty acid biosynthesis 418:440 These domains translocate into the nucleus, where they activate genes of cholesterol and fatty acid biosynthesis. 10419520 13 26 theme SREBPs 1835:1840 arg1 helix 1826:1830 the first transmembrane helix 1802:1830 the first transmembrane helix of SREBPs 1802:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 10 27 contain contains 1319:1326 arg2 residues 1360:1367 two potential zinc-coordinating residues 1328:1367 two potential zinc-coordinating residues 1328:1367 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 10 27 contain contains 1319:1326 arg1 sequence 1296:1303 The HEIGH sequence 1286:1303 The HEIGH sequence 1286:1303 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 4 28 theme ER 670:671 arg1 lumen 673:677 the ER lumen 666:677 the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain 666:825 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 10 29 theme zinc-coordinating 1342:1358 arg1 residues 1360:1367 two potential zinc-coordinating residues 1328:1367 two potential zinc-coordinating residues 1328:1367 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 6 30 theme site 979:982 arg1 mapping 984:990 glycosylation site mapping 965:990 glycosylation site mapping 965:990 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 2 31 theme fatty 418:422 arg1 biosynthesis 429:440 fatty acid biosynthesis 418:440 fatty acid biosynthesis 418:440 These domains translocate into the nucleus, where they activate genes of cholesterol and fatty acid biosynthesis. 10419520 12 32 from located 1600:1606 arg1 sequence 1625:1632 a hydrophobic sequence 1611:1632 a hydrophobic sequence 1611:1632 This residue, too, is located in a hydrophobic sequence. 10419520 9 33 theme S2P 1212:1214 arg1 sequences 1199:1207 the long hydrophilic sequences 1178:1207 the long hydrophilic sequences of S2P 1178:1214 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 6 34 theme glycosylation 965:977 arg1 mapping 984:990 glycosylation site mapping 965:990 glycosylation site mapping 965:990 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 0 35 theme S2P 21:23 arg1 protein 28:34 a protein 26:34 a protein required for intramembranous cleavage of sterol regulatory element-binding proteins 26:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 0 35 theme S2P 21:23 arg1 topology 9:16 Membrane topology 0:16 Membrane topology of S2P 0:23 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 12 36 located located 1600:1606 arg2 residue 1583:1589 This residue 1578:1589 This residue 1578:1589 This residue, too, is located in a hydrophobic sequence. 10419520 12 36 located located 1600:1606 arg1 sequence 1625:1632 a hydrophobic sequence 1611:1632 a hydrophobic sequence 1611:1632 This residue, too, is located in a hydrophobic sequence. 10419520 7 37 dep face 1078:1081 arg1 cytosol 1087:1093 the cytosol 1083:1093 S2P face the cytosol 1074:1093 Both the NH(2) and COOH termini of S2P face the cytosol. 10419520 12 38 from sequence 1625:1632 arg1 located 1600:1606 located 1600:1606 located 1600:1606 This residue, too, is located in a hydrophobic sequence. 10419520 9 39 gly glycosylated 1223:1234 arg1 sequences 1199:1207 the long hydrophilic sequences 1178:1207 the long hydrophilic sequences of S2P 1178:1214 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 9 39 gly glycosylated 1223:1234 arg1 three 1169:1173 three 1169:1173 three 1169:1173 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 13 40 theme Leu-Cys 1786:1792 arg1 target 1776:1781 its target 1772:1781 its target 1772:1781 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 40 theme Leu-Cys 1786:1792 arg1 bond 1794:1797 a Leu-Cys bond 1784:1797 a Leu-Cys bond in the first transmembrane helix of SREBPs 1784:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 2 41 theme cholesterol 402:412 arg1 genes 393:397 genes 393:397 genes of cholesterol and fatty acid biosynthesis 393:440 These domains translocate into the nucleus, where they activate genes of cholesterol and fatty acid biosynthesis. 10419520 9 42 theme long 1182:1185 arg1 sequences 1199:1207 the long hydrophilic sequences 1178:1207 the long hydrophilic sequences of S2P 1178:1214 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 3 43 theme hydrophilic 579:589 arg1 loop 591:594 a single hydrophilic loop 570:594 a single hydrophilic loop 570:594 The SREBPs are oriented in the membrane in a hairpin fashion, with the NH(2)- and COOH-terminal domains facing the cytosol and a single hydrophilic loop projecting into the lumen. 10419520 4 44 theme first 627:631 arg1 cleavage 633:640 The first cleavage 623:640 The first cleavage 623:640 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 13 45 theme ideal 1747:1751 arg1 position 1753:1760 an ideal position 1744:1760 an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs 1744:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 10 46 theme long 1392:1395 arg1 segment 1409:1415 a long hydrophobic segment 1390:1415 a long hydrophobic segment 1390:1415 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 9 47 theme hydrophilic 1187:1197 arg1 sequences 1199:1207 the long hydrophilic sequences 1178:1207 the long hydrophilic sequences of S2P 1178:1214 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 13 48 located located 1713:1719 arg1 membrane 1732:1739 the membrane 1728:1739 the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs 1728:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 48 located located 1713:1719 arg2 S2P 1706:1708 S2P 1706:1708 S2P 1706:1708 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 48 located located 1713:1719 arg2 located 1713:1719 located 1713:1719 located 1713:1719 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 48 located located 1713:1719 arg2 site 1698:1701 the active site 1687:1701 the active site of S2P 1687:1708 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 11 49 theme third 1525:1529 arg1 residue 1544:1550 the third coordinating residue 1521:1550 the third coordinating residue for the active site zinc 1521:1575 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 3 50 theme COOH-terminal 525:537 arg1 domains 539:545 COOH-terminal domains 525:545 COOH-terminal domains 525:545 The SREBPs are oriented in the membrane in a hairpin fashion, with the NH(2)- and COOH-terminal domains facing the cytosol and a single hydrophilic loop projecting into the lumen. 10419520 6 51 theme protease 941:948 arg1 protection 950:959 protease protection 941:959 protease protection 941:959 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 9 52 theme sequences 1199:1207 arg1 sequences 1199:1207 the long hydrophilic sequences 1178:1207 the long hydrophilic sequences of S2P 1178:1214 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 9 52 theme sequences 1199:1207 arg1 three 1169:1173 three 1169:1173 three 1169:1173 All three of the long hydrophilic sequences of S2P can be glycosylated, indicating that they all project into the lumen. 10419520 10 53 theme S2P 1308:1310 arg1 sequence 1296:1303 The HEIGH sequence 1286:1303 The HEIGH sequence 1286:1303 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 12 54 theme hydrophobic 1613:1623 arg1 sequence 1625:1632 a hydrophobic sequence 1611:1632 a hydrophobic sequence 1611:1632 This residue, too, is located in a hydrophobic sequence. 10419520 3 55 theme single 572:577 arg1 loop 591:594 a single hydrophilic loop 570:594 a single hydrophilic loop 570:594 The SREBPs are oriented in the membrane in a hairpin fashion, with the NH(2)- and COOH-terminal domains facing the cytosol and a single hydrophilic loop projecting into the lumen. 10419520 10 56 theme potential 1332:1340 arg1 residues 1360:1367 two potential zinc-coordinating residues 1328:1367 two potential zinc-coordinating residues 1328:1367 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 5 57 theme membrane 830:837 arg1 protein 839:845 A membrane protein 828:845 A membrane protein 828:845 A membrane protein, designated S2P, a putative zinc metalloprotease, is required for this cleavage. 10419520 2 58 theme biosynthesis 429:440 arg1 genes 393:397 genes 393:397 genes of cholesterol and fatty acid biosynthesis 393:440 These domains translocate into the nucleus, where they activate genes of cholesterol and fatty acid biosynthesis. 10419520 1 59 theme endoplasmic 301:311 arg1 ER 324:325 ER 324:325 ER 324:325 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 59 theme endoplasmic 301:311 arg1 reticulum 313:321 the endoplasmic reticulum 297:321 the endoplasmic reticulum (ER) 297:326 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 4 60 theme transmembrane 806:818 arg1 domain 820:825 the first transmembrane domain 796:825 the first transmembrane domain 796:825 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 7 61 theme S2P 1074:1076 arg1 face 1078:1081 S2P face the cytosol 1074:1093 S2P face the cytosol 1074:1093 Both the NH(2) and COOH termini of S2P face the cytosol. 10419520 6 62 theme ER 1025:1026 arg1 membranes 1028:1036 ER membranes 1025:1036 ER membranes 1025:1036 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 1 63 theme reticulum 313:321 arg1 membranes 284:292 membranes 284:292 membranes of the endoplasmic reticulum (ER) 284:326 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 6 64 gly glycosylation 965:977 arg2 site 979:982 glycosylation site mapping 965:990 glycosylation site mapping 965:990 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 6 64 gly glycosylation 965:977 arg2 mapping 984:990 glycosylation site mapping 965:990 glycosylation site mapping 965:990 Here, we use protease protection and glycosylation site mapping to define the topology of S2P in ER membranes. 10419520 10 65 theme hydrophobic 1397:1407 arg1 segment 1409:1415 a long hydrophobic segment 1390:1415 a long hydrophobic segment 1390:1415 The HEIGH sequence of S2P, which contains two potential zinc-coordinating residues, is contained within a long hydrophobic segment. 10419520 11 66 theme Aspartic 1418:1425 arg1 acid 1427:1430 Aspartic acid 467 1418:1434 Aspartic acid 467 1418:1434 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 1 67 theme sterol-depleted 124:138 arg1 cells 150:154 sterol-depleted mammalian cells 124:154 sterol-depleted mammalian cells 124:154 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 68 theme -terminal 206:214 arg1 proteins 261:268 sterol regulatory element-binding proteins 227:268 sterol regulatory element-binding proteins (SREBPs) 227:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 68 theme -terminal 206:214 arg1 domains 216:222 the NH(2)-terminal domains 197:222 the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) 197:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 13 69 theme first 1806:1810 arg1 helix 1826:1830 the first transmembrane helix 1802:1830 the first transmembrane helix of SREBPs 1802:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 11 70 theme coordinating 1531:1542 arg1 residue 1544:1550 the third coordinating residue 1521:1550 the third coordinating residue for the active site zinc 1521:1575 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 1 71 theme mammalian 140:148 arg1 cells 150:154 sterol-depleted mammalian cells 124:154 sterol-depleted mammalian cells 124:154 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 13 72 from membrane 1732:1739 arg1 position 1753:1760 an ideal position 1744:1760 an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs 1744:1840 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 0 73 theme intramembranous 49:63 arg1 cleavage 65:72 intramembranous cleavage 49:72 intramembranous cleavage of sterol regulatory element-binding proteins 49:118 Membrane topology of S2P, a protein required for intramembranous cleavage of sterol regulatory element-binding proteins. 10419520 13 74 theme S2P 1706:1708 arg1 located 1713:1719 located 1713:1719 located 1713:1719 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 74 theme S2P 1706:1708 arg1 S2P 1706:1708 S2P 1706:1708 S2P 1706:1708 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 74 theme S2P 1706:1708 arg1 site 1698:1701 the active site 1687:1701 the active site of S2P 1687:1708 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 3 75 dep NH 514:515 arg1 the 510:512 the 510:512 the 510:512 The SREBPs are oriented in the membrane in a hairpin fashion, with the NH(2)- and COOH-terminal domains facing the cytosol and a single hydrophilic loop projecting into the lumen. 10419520 11 76 theme active 1560:1565 arg1 zinc 1572:1575 the active site zinc 1556:1575 the active site zinc 1556:1575 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 4 77 attach released 728:735 arg1 membrane 746:753 the membrane 742:753 the membrane 742:753 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 4 77 attach released 728:735 arg2 intermediate 694:705 an intermediate 691:705 an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain 691:825 The first cleavage occurs at Site-1 within the ER lumen to generate an intermediate that is subsequently released from the membrane by cleavage at Site-2, which lies within the first transmembrane domain. 10419520 11 78 theme site 1567:1570 arg1 zinc 1572:1575 the active site zinc 1556:1575 the active site zinc 1556:1575 Aspartic acid 467, located approximately 300 residues away from the HEIGH sequence, appears to provide the third coordinating residue for the active site zinc. 10419520 3 79 theme hairpin 488:494 arg1 fashion 496:502 a hairpin fashion 486:502 a hairpin fashion 486:502 The SREBPs are oriented in the membrane in a hairpin fashion, with the NH(2)- and COOH-terminal domains facing the cytosol and a single hydrophilic loop projecting into the lumen. 10419520 13 80 theme active 1691:1696 arg1 located 1713:1719 located 1713:1719 located 1713:1719 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 80 theme active 1691:1696 arg1 S2P 1706:1708 S2P 1706:1708 S2P 1706:1708 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 13 80 theme active 1691:1696 arg1 site 1698:1701 the active site 1687:1701 the active site of S2P 1687:1708 The hydrophobicity of these sequences suggests that the active site of S2P is located within the membrane in an ideal position to cleave its target, a Leu-Cys bond in the first transmembrane helix of SREBPs. 10419520 1 81 theme sterol 227:232 arg1 SREBPs 271:276 SREBPs 271:276 SREBPs 271:276 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 10419520 1 81 theme sterol 227:232 arg1 proteins 261:268 sterol regulatory element-binding proteins 227:268 sterol regulatory element-binding proteins (SREBPs) 227:277 In sterol-depleted mammalian cells, a two-step proteolytic process releases the NH(2)-terminal domains of sterol regulatory element-binding proteins (SREBPs) from membranes of the endoplasmic reticulum (ER). 22451694 0 0 theme fibronectin 69:79 arg1 receptor 81:88 the fibronectin receptor 65:88 the fibronectin receptor 65:88 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 4 1 theme dynamics 687:694 arg1 simulation 696:705 a molecular dynamics simulation 675:705 a molecular dynamics simulation 675:705 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 3 2 theme antibody-bound 482:495 arg1 chain 500:504 The antibody-bound β1 chain 478:504 The antibody-bound β1 chain 478:504 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 4 3 theme molecule 830:837 arg1 site 805:808 the ligand-binding site 786:808 the ligand-binding site 786:808 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 4 3 theme molecule 830:837 arg1 rest 818:821 the rest 814:821 the rest of the molecule 814:837 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 4 3 theme molecule 830:837 arg1 molecule 830:837 the molecule 826:837 the molecule 826:837 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 1 4 theme extracellular 142:154 arg1 fibronectin 171:181 the extracellular matrix protein fibronectin 138:181 the extracellular matrix protein fibronectin 138:181 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 2 5 theme inhibitory 325:334 arg1 antibody 336:343 an allosteric inhibitory antibody 311:343 an allosteric inhibitory antibody 311:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 5 6 theme devoid 945:950 arg1 α5β1 940:943 α5β1 940:943 α5β1 devoid of glycosylation sites 940:973 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 2 7 theme headpiece 283:291 arg1 fragment 293:300 the α5β1 integrin headpiece fragment 265:300 the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody 265:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 3 8 theme limited 544:550 arg1 changes 563:569 very limited structural changes 539:569 very limited structural changes 539:569 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 0 9 theme receptor 81:88 arg1 details 54:60 atomic details 47:60 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 0:89 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 5 10 theme top 928:930 arg1 face 932:935 the top face 924:935 the top face of α5β1 devoid of glycosylation sites 924:973 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 2 11 theme integrin 274:281 arg1 fragment 293:300 the α5β1 integrin headpiece fragment 265:300 the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody 265:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 2 12 theme allosteric 314:323 arg1 antibody 336:343 an allosteric inhibitory antibody 311:343 an allosteric inhibitory antibody 311:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 4 13 theme molecular 677:685 arg1 simulation 696:705 a molecular dynamics simulation 675:705 a molecular dynamics simulation 675:705 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 2 14 contain containing 435:444 arg1 peptide 427:433 a ligand peptide 418:433 a ligand peptide containing the Arg-Gly-Asp (RGD) sequence 418:475 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 2 14 contain containing 435:444 arg2 sequence 468:475 the Arg-Gly-Asp (RGD) sequence 446:475 the Arg-Gly-Asp (RGD) sequence 446:475 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 2 15 theme α5β1 269:272 arg1 fragment 293:300 the α5β1 integrin headpiece fragment 265:300 the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody 265:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 1 16 theme matrix 156:161 arg1 fibronectin 171:181 the extracellular matrix protein fibronectin 138:181 the extracellular matrix protein fibronectin 138:181 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 2 17 theme peptide 427:433 arg1 presence 406:413 presence 406:413 presence 406:413 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 2 17 theme peptide 427:433 arg1 absence 394:400 absence 394:400 absence 394:400 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 6 18 theme residue 1042:1048 arg1 prediction 1015:1024 the precise prediction 1003:1024 the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin 1003:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 1 19 theme protein 163:169 arg1 fibronectin 171:181 the extracellular matrix protein fibronectin 138:181 the extracellular matrix protein fibronectin 138:181 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 6 20 theme acceptor 1033:1040 arg1 residue 1042:1048 the acceptor residue 1029:1048 the acceptor residue 1029:1048 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 3 21 theme structural 552:561 arg1 changes 563:569 very limited structural changes 539:569 very limited structural changes 539:569 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 0 22 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 0:89 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 4 23 from role 731:734 arg1 coupling 769:776 the conformational coupling 750:776 the conformational coupling between the ligand-binding site and the rest of the molecule 750:837 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 4 24 theme important 721:729 arg1 role 731:734 an important role 718:734 an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule 718:837 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 5 25 theme RGD-binding 844:854 arg1 situated 866:873 situated 866:873 situated 866:873 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 5 25 theme RGD-binding 844:854 arg1 pocket 856:861 The RGD-binding pocket 840:861 The RGD-binding pocket 840:861 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 0 26 theme α5β1 21:24 arg1 ectodomain 35:44 α5β1 integrin ectodomain 21:44 α5β1 integrin ectodomain 21:44 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 0 27 dep structure 8:16 arg1 details 54:60 atomic details 47:60 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 0:89 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 5 28 from center 882:887 arg1 situated 866:873 situated 866:873 situated 866:873 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 5 28 from center 882:887 arg1 pocket 856:861 The RGD-binding pocket 840:861 The RGD-binding pocket 840:861 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 1 29 theme Integrin 91:98 arg1 receptor 125:132 a major cellular receptor 108:132 a major cellular receptor for the extracellular matrix protein fibronectin 108:181 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 1 29 theme Integrin 91:98 arg1 α5β1 100:103 Integrin α5β1 91:103 Integrin α5β1 91:103 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 5 30 theme sites 969:973 arg1 devoid 945:950 devoid 945:950 devoid 945:950 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 6 31 theme precise 1007:1013 arg1 prediction 1015:1024 the precise prediction 1003:1024 the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin 1003:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 32 theme synergy 1068:1074 arg1 site 1076:1079 the auxiliary synergy site 1054:1079 the auxiliary synergy site of fibronectin 1054:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 32 theme synergy 1068:1074 arg1 fibronectin 1084:1094 fibronectin 1084:1094 fibronectin 1084:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 2 33 theme fragment 293:300 arg1 structure 252:260 A crystal structure 242:260 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody 242:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 5 34 gly glycosylation 955:967 arg2 sites 969:973 glycosylation sites 955:973 glycosylation sites 955:973 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 6 35 theme α5 1103:1104 arg1 subunit 1106:1112 the α5 subunit 1099:1112 the α5 subunit 1099:1112 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 0 36 theme ectodomain 35:44 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 0:89 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 4 37 theme ligand-binding 790:803 arg1 site 805:808 the ligand-binding site 786:808 the ligand-binding site 786:808 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 4 37 theme ligand-binding 790:803 arg1 molecule 830:837 the molecule 826:837 the molecule 826:837 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 1 38 theme fundamental 195:205 arg1 role 207:210 a fundamental role 193:210 a fundamental role 193:210 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 0 39 theme integrin 26:33 arg1 ectodomain 35:44 α5β1 integrin ectodomain 21:44 α5β1 integrin ectodomain 21:44 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 3 40 theme cell 612:615 arg1 adhesion 617:624 cell adhesion 612:624 cell adhesion mediated by nonactivated integrins 612:659 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 2 41 theme ligand 420:425 arg1 peptide 427:433 a ligand peptide 418:433 a ligand peptide containing the Arg-Gly-Asp (RGD) sequence 418:475 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 2 42 theme Arg-Gly-Asp 450:460 arg1 sequence 468:475 the Arg-Gly-Asp (RGD) sequence 446:475 the Arg-Gly-Asp (RGD) sequence 446:475 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 3 43 theme adhesion 617:624 arg1 step 604:607 the initial step 592:607 the initial step of cell adhesion mediated by nonactivated integrins 592:659 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 6 44 theme binding 1177:1183 arg1 assays 1185:1190 kinetic binding assays 1169:1190 kinetic binding assays 1169:1190 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 45 theme auxiliary 1058:1066 arg1 site 1076:1079 the auxiliary synergy site 1054:1079 the auxiliary synergy site of fibronectin 1054:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 45 theme auxiliary 1058:1066 arg1 fibronectin 1084:1094 fibronectin 1084:1094 fibronectin 1084:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 46 theme fibronectin 1084:1094 arg1 site 1076:1079 the auxiliary synergy site 1054:1079 the auxiliary synergy site of fibronectin 1054:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 46 theme fibronectin 1084:1094 arg1 fibronectin 1084:1094 fibronectin 1084:1094 fibronectin 1084:1094 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 6 47 theme kinetic 1169:1175 arg1 assays 1185:1190 kinetic binding assays 1169:1190 kinetic binding assays 1169:1190 The structure also enabled the precise prediction of the acceptor residue for the auxiliary synergy site of fibronectin on the α5 subunit, which was experimentally confirmed by mutagenesis and kinetic binding assays. 22451694 3 48 theme β1 497:498 arg1 chain 500:504 The antibody-bound β1 chain 478:504 The antibody-bound β1 chain 478:504 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 5 49 theme glycosylation 955:967 arg1 sites 969:973 glycosylation sites 955:973 glycosylation sites 955:973 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 5 50 theme trenchlike 894:903 arg1 center 882:887 the center 878:887 the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites 878:973 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 5 51 theme α5β1 940:943 arg1 face 932:935 the top face 924:935 the top face of α5β1 devoid of glycosylation sites 924:973 The RGD-binding pocket is situated at the center of a trenchlike exposed surface on the top face of α5β1 devoid of glycosylation sites. 22451694 4 52 theme conformational 754:767 arg1 coupling 769:776 the conformational coupling 750:776 the conformational coupling between the ligand-binding site and the rest of the molecule 750:837 Furthermore, a molecular dynamics simulation pointed to an important role for Ca(2+) in the conformational coupling between the ligand-binding site and the rest of the molecule. 22451694 2 53 dep absence 394:400 arg1 the 390:392 the 390:392 the 390:392 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 1 54 theme major 110:114 arg1 receptor 125:132 a major cellular receptor 108:132 a major cellular receptor for the extracellular matrix protein fibronectin 108:181 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 1 54 theme major 110:114 arg1 α5β1 100:103 Integrin α5β1 91:103 Integrin α5β1 91:103 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 3 55 theme RGD 523:525 arg1 ligand 527:532 the RGD ligand 519:532 the RGD ligand 519:532 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 3 56 theme nonactivated 638:649 arg1 integrins 651:659 nonactivated integrins 638:659 nonactivated integrins 638:659 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 22451694 2 57 dep resolution 371:380 arg1 both 382:385 both 382:385 both 382:385 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 1 58 theme cellular 116:123 arg1 receptor 125:132 a major cellular receptor 108:132 a major cellular receptor for the extracellular matrix protein fibronectin 108:181 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 1 58 theme cellular 116:123 arg1 α5β1 100:103 Integrin α5β1 91:103 Integrin α5β1 91:103 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 1 59 theme mammalian 219:227 arg1 development 229:239 mammalian development 219:239 mammalian development 219:239 Integrin α5β1 is a major cellular receptor for the extracellular matrix protein fibronectin and plays a fundamental role during mammalian development. 22451694 2 60 theme crystal 244:250 arg1 structure 252:260 A crystal structure 242:260 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody 242:343 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 2 61 theme 2.9-Å 365:369 arg1 resolution 371:380 a 2.9-Å resolution 363:380 a 2.9-Å resolution 363:380 A crystal structure of the α5β1 integrin headpiece fragment bound by an allosteric inhibitory antibody was determined at a 2.9-Å resolution both in the absence and presence of a ligand peptide containing the Arg-Gly-Asp (RGD) sequence. 22451694 0 62 theme atomic 47:52 arg1 details 54:60 atomic details 47:60 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 0:89 Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. 22451694 3 63 theme initial 596:602 arg1 step 604:607 the initial step 592:607 the initial step of cell adhesion mediated by nonactivated integrins 592:659 The antibody-bound β1 chain accommodated the RGD ligand with very limited structural changes, which may represent the initial step of cell adhesion mediated by nonactivated integrins. 9524075 12 0 theme S 1440:1440 arg1 molecule 1442:1449 the cathepsin S molecule 1426:1449 the cathepsin S molecule 1426:1449 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 12 1 theme plasma 1352:1357 arg1 membrane 1359:1366 the plasma membrane 1348:1366 the plasma membrane 1348:1366 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 11 2 theme non-glycosylated 1245:1260 arg1 S 1275:1275 non-glycosylated procathepsin S 1245:1275 non-glycosylated procathepsin S 1245:1275 Subcellular fractionation showed non-glycosylated procathepsin S in the membrane fraction. 9524075 6 3 contain contained 663:671 arg1 type 643:646 Secreted wild type 629:646 Secreted wild type procathepsin S 629:661 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 6 3 contain contained 663:671 arg1 chain 712:716 the oligosaccharide chain 692:716 the oligosaccharide chain 692:716 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 6 3 contain contained 663:671 arg2 Man-6-phosphate 673:687 Man-6-phosphate 673:687 Man-6-phosphate 673:687 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 6 3 contain contained 663:671 arg2 type 643:646 Secreted wild type 629:646 Secreted wild type procathepsin S 629:661 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 12 4 theme Man-6-phosphate 1463:1477 arg1 residue 1479:1485 the Man-6-phosphate residue 1459:1485 the Man-6-phosphate residue 1459:1485 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 3 5 theme human 323:327 arg1 S 342:342 human procathepsin S 323:342 human procathepsin S 323:342 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 11 6 theme membrane 1284:1291 arg1 fraction 1293:1300 the membrane fraction 1280:1300 the membrane fraction 1280:1300 Subcellular fractionation showed non-glycosylated procathepsin S in the membrane fraction. 9524075 5 7 theme precursors 558:567 arg1 amounts 543:549 Large amounts 537:549 Large amounts of the precursors 537:567 Large amounts of the precursors were secreted into the culture media by both transfectants. 9524075 5 7 theme precursors 558:567 arg1 precursors 558:567 the precursors 554:567 the precursors 554:567 Large amounts of the precursors were secreted into the culture media by both transfectants. 9524075 12 8 from motif 1417:1421 arg1 molecule 1442:1449 the cathepsin S molecule 1426:1449 the cathepsin S molecule 1426:1449 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 8 9 theme In 820:821 arg1 processing 829:838 In vitro processing 820:838 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S 820:900 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 3 10 theme procathepsin 329:340 arg1 S 342:342 human procathepsin S 323:342 human procathepsin S 323:342 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 8 11 theme procathepsin 887:898 arg1 S 900:900 non-glycosylated procathepsin S 870:900 non-glycosylated procathepsin S 870:900 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 8 12 theme non-glycosylated 870:885 arg1 S 900:900 non-glycosylated procathepsin S 870:900 non-glycosylated procathepsin S 870:900 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 9 13 theme S 1078:1078 arg1 reuptake 1015:1022 A reuptake 1013:1022 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells 1013:1095 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 3 14 theme wild 313:316 arg1 type 318:321 either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln 306:450 type 318:321 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 3 15 theme HEK 229:231 arg1 293-cells 233:241 HEK 293-cells 229:241 HEK 293-cells which do not express cathepsin S 229:274 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 6 16 dep type 643:646 arg1 S 661:661 procathepsin S 648:661 Secreted wild type procathepsin S 629:661 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 1 17 theme Cathepsin 69:77 arg1 protease 103:110 a lysosomal cysteine protease 82:110 a lysosomal cysteine protease 82:110 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 1 17 theme Cathepsin 69:77 arg1 S 79:79 Cathepsin S 69:79 Cathepsin S 69:79 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 1 17 theme Cathepsin 69:77 arg1 precursor 140:148 inactive precursor 131:148 inactive precursor 131:148 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 9 18 theme non-glycosylated 1048:1063 arg1 S 1078:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 3 19 theme type 318:321 arg1 cDNA 298:301 cDNA 298:301 cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln 298:450 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 12 20 theme cathepsin 1430:1438 arg1 molecule 1442:1449 the cathepsin S molecule 1426:1449 the cathepsin S molecule 1426:1449 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 3 21 from proregion 422:430 arg1 Asn 380:382 Asn 380:382 Asn of the only glycosylation site in the proregion 380:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 12 22 theme additional 1398:1407 arg1 motif 1417:1421 an additional sorting motif 1395:1421 an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue 1395:1485 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 9 23 theme procathepsin 1065:1076 arg1 S 1078:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 3 24 theme only 391:394 arg1 site 410:413 the only glycosylation site 387:413 the only glycosylation site in the proregion 387:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 5 25 theme culture 592:598 arg1 media 600:604 the culture media 588:604 the culture media 588:604 Large amounts of the precursors were secreted into the culture media by both transfectants. 9524075 5 26 theme Large 537:541 arg1 amounts 543:549 Large amounts 537:549 Large amounts of the precursors 537:567 Large amounts of the precursors were secreted into the culture media by both transfectants. 9524075 5 26 theme Large 537:541 arg1 precursors 558:567 the precursors 554:567 the precursors 554:567 Large amounts of the precursors were secreted into the culture media by both transfectants. 9524075 12 27 theme degrees 1373:1379 arg1 C 1381:1381 2 degrees C 1371:1381 2 degrees C 1371:1381 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 8 28 theme glycosylated 843:854 arg1 processing 829:838 In vitro processing 820:838 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S 820:900 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 12 29 theme procathepsin 1320:1331 arg1 S 1333:1333 Non-glycosylated procathepsin S 1303:1333 Non-glycosylated procathepsin S 1303:1333 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 3 30 theme mutant 349:354 arg1 S 369:369 a mutant procathepsin S 347:369 a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln 347:450 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 10 31 theme Small 1116:1120 arg1 S 1150:1150 mature cathepsin S 1133:1150 mature cathepsin S 1133:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 10 31 theme Small 1116:1120 arg1 amounts 1122:1128 Small amounts 1116:1128 Small amounts of mature cathepsin S 1116:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 6 32 theme procathepsin 648:659 arg1 S 661:661 procathepsin S 648:661 Secreted wild type procathepsin S 629:661 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 8 33 theme proper 997:1002 arg1 folding 1004:1010 proper folding 997:1010 proper folding 997:1010 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 0 34 theme human 28:32 arg1 procathepsin 34:45 non-glycosylated human procathepsin 11:45 non-glycosylated human procathepsin 11:45 Sorting of non-glycosylated human procathepsin S in mammalian cells. 9524075 4 35 theme procathepsin 507:518 arg1 S 520:520 procathepsin S 507:520 procathepsin S 507:520 The cells expressed glycosylated and non-glycosylated procathepsin S, respectively. 9524075 9 36 gly non-glycosylated 1048:1063 arg1 S 1078:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 2 37 theme proteolytic 189:199 arg1 cleavage 201:208 a proteolytic cleavage 187:208 a proteolytic cleavage of the propeptide 187:226 It is activated in the lysosomes by a proteolytic cleavage of the propeptide. 9524075 1 38 theme lysosomal 84:92 arg1 protease 103:110 a lysosomal cysteine protease 82:110 a lysosomal cysteine protease 82:110 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 1 38 theme lysosomal 84:92 arg1 S 79:79 Cathepsin S 69:79 Cathepsin S 69:79 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 7 39 theme culture 805:811 arg1 media 813:817 the culture media 801:817 the culture media 801:817 Wild type procathepsin S was activated in the cells but no maturation occurred in the culture media. 9524075 0 40 theme non-glycosylated 11:26 arg1 procathepsin 34:45 non-glycosylated human procathepsin 11:45 non-glycosylated human procathepsin 11:45 Sorting of non-glycosylated human procathepsin S in mammalian cells. 9524075 11 41 theme Subcellular 1212:1222 arg1 fractionation 1224:1236 Subcellular fractionation 1212:1236 Subcellular fractionation 1212:1236 Subcellular fractionation showed non-glycosylated procathepsin S in the membrane fraction. 9524075 1 42 theme cysteine 94:101 arg1 protease 103:110 a lysosomal cysteine protease 82:110 a lysosomal cysteine protease 82:110 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 1 42 theme cysteine 94:101 arg1 S 79:79 Cathepsin S 69:79 Cathepsin S 69:79 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 6 43 theme wild 638:641 arg1 type 643:646 Secreted wild type 629:646 Secreted wild type procathepsin S 629:661 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 8 44 dep In 820:821 arg1 vitro 823:827 vitro 823:827 vitro 823:827 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 11 45 theme procathepsin 1262:1273 arg1 S 1275:1275 non-glycosylated procathepsin S 1245:1275 non-glycosylated procathepsin S 1245:1275 Subcellular fractionation showed non-glycosylated procathepsin S in the membrane fraction. 9524075 6 46 theme Secreted 629:636 arg1 type 643:646 Secreted wild type 629:646 Secreted wild type procathepsin S 629:661 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 3 47 theme glycosylation 396:408 arg1 site 410:413 the only glycosylation site 387:413 the only glycosylation site in the proregion 387:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 0 48 theme procathepsin 34:45 arg1 Sorting 0:6 Sorting 0:6 Sorting of non-glycosylated human procathepsin 0:45 Sorting of non-glycosylated human procathepsin S in mammalian cells. 9524075 10 49 theme cathepsin 1140:1148 arg1 S 1150:1150 mature cathepsin S 1133:1150 mature cathepsin S 1133:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 12 50 gly Non-glycosylated 1303:1318 arg1 S 1333:1333 Non-glycosylated procathepsin S 1303:1333 Non-glycosylated procathepsin S 1303:1333 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 8 51 theme S 900:900 arg1 processing 829:838 In vitro processing 820:838 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S 820:900 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 3 52 theme site 410:413 arg1 Asn 380:382 Asn 380:382 Asn of the only glycosylation site in the proregion 380:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 0 53 theme mammalian 52:60 arg1 cells 62:66 mammalian cells 52:66 mammalian cells 52:66 Sorting of non-glycosylated human procathepsin S in mammalian cells. 9524075 10 54 theme mature 1133:1138 arg1 S 1150:1150 mature cathepsin S 1133:1150 mature cathepsin S 1133:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 9 55 gly glycosylated 1031:1042 arg1 S 1078:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 10 56 theme S 1150:1150 arg1 S 1150:1150 mature cathepsin S 1133:1150 mature cathepsin S 1133:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 10 56 theme S 1150:1150 arg1 amounts 1122:1128 Small amounts 1116:1128 Small amounts of mature cathepsin S 1116:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 0 57 gly non-glycosylated 11:26 arg1 procathepsin 34:45 non-glycosylated human procathepsin 11:45 non-glycosylated human procathepsin 11:45 Sorting of non-glycosylated human procathepsin S in mammalian cells. 9524075 3 58 from site 410:413 arg1 proregion 422:430 the proregion 418:430 the proregion 418:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 3 59 theme S 369:369 arg1 cDNA 298:301 cDNA 298:301 cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln 298:450 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 3 60 dep type 318:321 arg1 S 342:342 human procathepsin S 323:342 human procathepsin S 323:342 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 3 61 gly glycosylation 396:408 arg2 site 410:413 the only glycosylation site 387:413 the only glycosylation site in the proregion 387:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 3 62 theme cathepsin 264:272 arg1 S 274:274 cathepsin S 264:274 cathepsin S 264:274 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 10 63 located detected 1157:1164 arg1 lysosomes 1173:1181 the lysosomes 1169:1181 the lysosomes of the mutant transfectants 1169:1209 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 10 63 located detected 1157:1164 arg2 amounts 1122:1128 Small amounts 1116:1128 Small amounts of mature cathepsin S 1116:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 10 63 located detected 1157:1164 arg2 S 1150:1150 mature cathepsin S 1133:1150 mature cathepsin S 1133:1150 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 12 64 theme sorting 1409:1415 arg1 motif 1417:1421 an additional sorting motif 1395:1421 an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue 1395:1485 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 8 65 theme oligosaccharide 953:967 arg1 chain 969:973 the oligosaccharide chain 949:973 the oligosaccharide chain 949:973 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 8 65 theme oligosaccharide 953:967 arg1 necessary 983:991 necessary 983:991 necessary 983:991 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 11 66 gly non-glycosylated 1245:1260 arg1 S 1275:1275 non-glycosylated procathepsin S 1245:1275 non-glycosylated procathepsin S 1245:1275 Subcellular fractionation showed non-glycosylated procathepsin S in the membrane fraction. 9524075 7 67 theme procathepsin 729:740 arg1 S 742:742 Wild type procathepsin S 719:742 Wild type procathepsin S 719:742 Wild type procathepsin S was activated in the cells but no maturation occurred in the culture media. 9524075 8 68 gly non-glycosylated 870:885 arg1 S 900:900 non-glycosylated procathepsin S 870:900 non-glycosylated procathepsin S 870:900 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 3 69 from Asn 380:382 arg1 proregion 422:430 the proregion 418:430 the proregion 418:430 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 9 70 theme HEK 1083:1085 arg1 293-cells 1087:1095 HEK 293-cells 1083:1095 HEK 293-cells 1083:1095 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 10 71 theme transfectants 1197:1209 arg1 lysosomes 1173:1181 the lysosomes 1169:1181 the lysosomes of the mutant transfectants 1169:1209 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 12 72 theme Non-glycosylated 1303:1318 arg1 S 1333:1333 Non-glycosylated procathepsin S 1303:1333 Non-glycosylated procathepsin S 1303:1333 Non-glycosylated procathepsin S was bound to the plasma membrane at 2 degrees C, suggesting an additional sorting motif in the cathepsin S molecule besides the Man-6-phosphate residue. 9524075 9 73 theme glycosylated 1031:1042 arg1 S 1078:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 the glycosylated and non-glycosylated procathepsin S 1027:1078 A reuptake of the glycosylated and non-glycosylated procathepsin S by HEK 293-cells could be observed. 9524075 8 74 theme active 913:918 arg1 enzymes 920:926 fully active enzymes 907:926 fully active enzymes 907:926 In vitro processing of glycosylated as well as of non-glycosylated procathepsin S gave fully active enzymes thus indicating that the oligosaccharide chain was not necessary for proper folding. 9524075 10 75 theme mutant 1190:1195 arg1 transfectants 1197:1209 the mutant transfectants 1186:1209 the mutant transfectants 1186:1209 Small amounts of mature cathepsin S were detected in the lysosomes of the mutant transfectants. 9524075 7 76 theme Wild 719:722 arg1 S 742:742 Wild type procathepsin S 719:742 Wild type procathepsin S 719:742 Wild type procathepsin S was activated in the cells but no maturation occurred in the culture media. 9524075 3 77 theme procathepsin 356:367 arg1 S 369:369 a mutant procathepsin S 347:369 a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln 347:450 HEK 293-cells which do not express cathepsin S were transfected with cDNA of either wild type human procathepsin S or a mutant procathepsin S in which Asn of the only glycosylation site in the proregion was replaced by Gln. 9524075 4 78 dep non-glycosylated 490:505 arg1 S 520:520 procathepsin S 507:520 procathepsin S 507:520 The cells expressed glycosylated and non-glycosylated procathepsin S, respectively. 9524075 2 79 theme propeptide 217:226 arg1 cleavage 201:208 a proteolytic cleavage 187:208 a proteolytic cleavage of the propeptide 187:226 It is activated in the lysosomes by a proteolytic cleavage of the propeptide. 9524075 6 80 theme oligosaccharide 696:710 arg1 chain 712:716 the oligosaccharide chain 692:716 the oligosaccharide chain 692:716 Secreted wild type procathepsin S contained Man-6-phosphate in the oligosaccharide chain. 9524075 1 81 theme inactive 131:138 arg1 S 79:79 Cathepsin S 69:79 Cathepsin S 69:79 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 1 81 theme inactive 131:138 arg1 precursor 140:148 inactive precursor 131:148 inactive precursor 131:148 Cathepsin S, a lysosomal cysteine protease, is synthesized as inactive precursor. 9524075 7 82 theme type 724:727 arg1 S 742:742 Wild type procathepsin S 719:742 Wild type procathepsin S 719:742 Wild type procathepsin S was activated in the cells but no maturation occurred in the culture media. 20879038 6 0 theme Gln 1420:1422 arg1 glycosylation 1371:1383 glycosylation 1371:1383 glycosylation 1371:1383 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 6 0 theme Gln 1420:1422 arg1 conversion 1406:1415 conversion 1406:1415 conversion of Gln to pyro-Glu 1406:1434 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 6 0 theme Gln 1420:1422 arg1 phosphorylation 1386:1400 phosphorylation 1386:1400 phosphorylation 1386:1400 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 5 1 from two 942:944 arg1 bPRP2 949:953 bPRP2 949:953 bPRP2 949:953 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 2 from Diabetic 630:637 arg1 subjects 607:614 20 different subjects 594:614 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 594:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 2 from Diabetic 630:637 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 3 dep identify 692:699 arg1 iv 974:975 iv 974:975 iv 974:975 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 4 4 theme distinct 484:491 arg1 isoforms 493:500 distinct isoforms 484:500 distinct isoforms 484:500 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 5 5 from Control 621:627 arg1 subjects 607:614 20 different subjects 594:614 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 594:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 5 from Control 621:627 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 6 dep possible 680:687 arg1 identify 692:699 identify 692:699 to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3 689:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 2 7 from PRP 305:307 arg1 PRP1 345:348 basic PRP1 339:348 basic PRP1 339:348 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 7 from PRP 305:307 arg1 2 354:354 2 354:354 2 354:354 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 5 8 from two 959:961 arg1 bPRP3 966:970 bPRP3 966:970 bPRP3 966:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 3 9 from Ser17 395:399 arg1 presence 361:368 The presence 357:368 The presence of a glucoronyl group at Ser17 357:399 The presence of a glucoronyl group at Ser17 was also noticed on aPRP. 20879038 7 10 theme Cancer 1573:1578 arg1 patients 1580:1587 the Head and Neck Cancer patients 1555:1587 the Head and Neck Cancer patients 1555:1587 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 3 11 attach presence 361:368 arg1 Ser17 395:399 Ser17 395:399 Ser17 395:399 The presence of a glucoronyl group at Ser17 was also noticed on aPRP. 20879038 3 11 attach presence 361:368 arg2 group 386:390 a glucoronyl group 373:390 a glucoronyl group 373:390 The presence of a glucoronyl group at Ser17 was also noticed on aPRP. 20879038 5 12 dep species 715:721 arg1 sites 921:925 (iii) other terminal monosaccharide sites 885:925 (iii) other terminal monosaccharide sites 885:925 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 six 928:930 six 928:930 six 928:930 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 species 715:721 the following species 701:721 the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3 701:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 two 959:961 two 959:961 two 959:961 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 sites 848:852 (ii) O-glycosylation sites 827:852 (ii) O-glycosylation sites 827:852 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 two 855:857 two 855:857 two 855:857 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 sites 744:748 (i) N-glycosylation sites 724:748 the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3 701:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 two 942:944 two 942:944 two 942:944 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 12 dep species 715:721 arg1 two 751:753 two 751:753 two 751:753 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 13 from Head 644:647 arg1 subjects 607:614 20 different subjects 594:614 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 594:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 13 from Head 644:647 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 14 from one 1137:1139 arg1 bPRP2 1144:1148 bPRP2 1144:1148 bPRP2 1144:1148 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 15 dep modifications 984:996 arg1 two 1027:1029 two 1027:1029 two 1027:1029 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 2 16 theme serine 211:216 arg1 phosphorylation 218:232 only the serine phosphorylation 202:232 only the serine phosphorylation at positions 8, 17, and 22 202:259 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 16 theme serine 211:216 arg1 date 196:199 date 196:199 date 196:199 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 17 located observed 286:293 arg1 PRP 305:307 acidic PRP 298:307 acidic PRP (aPRPs) 298:315 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 17 located observed 286:293 arg2 Up 190:191 Up 190:191 Up to date, only the serine phosphorylation at positions 8, 17, and 22 190:259 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 17 located observed 286:293 arg1 aPRPs 310:314 aPRPs 310:314 aPRPs 310:314 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 1 18 theme complex 121:127 arg1 family 129:134 the most complex family 112:134 the most complex family of salivary peptides with distinct isoforms and PTMs 112:187 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 1 18 theme complex 121:127 arg1 proteins 92:99 Proline-rich proteins 79:99 Proline-rich proteins (PRPs) 79:106 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 5 19 from one 1151:1153 arg1 bPRP3 1158:1162 bPRP3 1158:1162 bPRP3 1158:1162 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 20 theme subjects 607:614 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 21 from Neck 653:656 arg1 subjects 607:614 20 different subjects 594:614 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 594:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 21 from Neck 653:656 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 1 22 theme peptides 148:155 arg1 family 129:134 the most complex family 112:134 the most complex family of salivary peptides with distinct isoforms and PTMs 112:187 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 1 22 theme peptides 148:155 arg1 proteins 92:99 Proline-rich proteins 79:99 Proline-rich proteins (PRPs) 79:106 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 5 23 from one 872:874 arg1 aPRP 879:882 aPRP 879:882 aPRP 879:882 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 24 theme monosaccharide 906:919 arg1 sites 921:925 (iii) other terminal monosaccharide sites 885:925 (iii) other terminal monosaccharide sites 885:925 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 25 theme basic 758:762 arg1 one 813:815 one 813:815 one 813:815 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 25 theme basic 758:762 arg1 protein 777:783 basic proline-rich protein 2 758:785 basic proline-rich protein 2 (bPRP2) 758:793 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 25 theme basic 758:762 arg1 bPRP2 788:792 bPRP2 788:792 bPRP2 788:792 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 25 theme basic 758:762 arg1 one 796:798 one 796:798 one 796:798 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 26 from one 1168:1170 arg1 aPRP1 1175:1179 aPRP1 1175:1179 aPRP1 1175:1179 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 6 27 theme new 1330:1332 arg1 residues 1347:1354 45 new PRP-modified residues 1327:1354 45 new PRP-modified residues 1327:1354 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 1 28 theme distinct 162:169 arg1 isoforms 171:178 distinct isoforms 162:178 distinct isoforms 162:178 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 7 29 theme hexosamine 1519:1528 arg1 modification 1530:1541 N-acetyl hexosamine modification 1510:1541 N-acetyl hexosamine modification 1510:1541 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 1 30 theme Proline-rich 79:90 arg1 proteins 92:99 Proline-rich proteins 79:99 Proline-rich proteins (PRPs) 79:106 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 1 30 theme Proline-rich 79:90 arg1 family 129:134 the most complex family 112:134 the most complex family of salivary peptides with distinct isoforms and PTMs 112:187 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 1 30 theme Proline-rich 79:90 arg1 PRPs 102:105 PRPs 102:105 PRPs 102:105 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 5 31 theme other 891:895 arg1 sites 921:925 (iii) other terminal monosaccharide sites 885:925 (iii) other terminal monosaccharide sites 885:925 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 2 32 from positions 237:245 arg1 phosphorylation 218:232 only the serine phosphorylation 202:232 only the serine phosphorylation at positions 8, 17, and 22 202:259 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 32 from positions 237:245 arg1 date 196:199 date 196:199 date 196:199 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 5 33 from nine 1074:1077 arg1 bPRP4 1082:1086 bPRP4 1082:1086 bPRP4 1082:1086 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 4 34 theme study 449:453 arg1 goal 436:439 The main goal 427:439 The main goal of this study 427:453 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 6 35 gly glycosylation 1371:1383 arg1 Gln 1420:1422 Gln 1420:1422 Gln 1420:1422 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 3 36 theme group 386:390 arg1 presence 361:368 The presence 357:368 The presence of a glucoronyl group at Ser17 357:399 The presence of a glucoronyl group at Ser17 was also noticed on aPRP. 20879038 0 37 theme posttranslational 12:28 arg1 modifications 30:42 new posttranslational modifications 8:42 new posttranslational modifications 8:42 Finding new posttranslational modifications in salivary proline-rich proteins. 20879038 5 38 dep sites 921:925 arg1 iii 886:888 iii 886:888 iii 886:888 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 39 from six 928:930 arg1 bPRP1 935:939 bPRP1 935:939 bPRP1 935:939 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 4 40 theme main 431:434 arg1 goal 436:439 The main goal 427:439 The main goal of this study 427:453 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 5 41 theme Cancer 658:663 arg1 groups 665:670 Cancer groups 658:670 Cancer groups 658:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 0 42 theme proline-rich 56:67 arg1 proteins 69:76 salivary proline-rich proteins 47:76 salivary proline-rich proteins 47:76 Finding new posttranslational modifications in salivary proline-rich proteins. 20879038 7 43 theme subject 1461:1467 arg1 groups 1469:1474 all subject groups 1457:1474 all subject groups 1457:1474 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 6 44 theme residues 1347:1354 arg1 identification 1309:1322 the identification 1305:1322 the identification of 45 new PRP-modified residues 1305:1354 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 5 45 dep two 1027:1029 arg1 eight 1055:1059 eight 1055:1059 eight 1055:1059 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 45 dep two 1027:1029 arg1 six 1041:1043 six 1041:1043 six 1041:1043 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 45 dep two 1027:1029 arg1 nine 1074:1077 nine 1074:1077 nine 1074:1077 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 46 from phosphorylation 1094:1108 arg1 one 1151:1153 one 1151:1153 one 1151:1153 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 46 from phosphorylation 1094:1108 arg1 one 1137:1139 one 1137:1139 one 1137:1139 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 46 from phosphorylation 1094:1108 arg1 serine 1113:1118 serine 1113:1118 serine 1113:1118 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 46 from phosphorylation 1094:1108 arg1 one 1168:1170 one 1168:1170 one 1168:1170 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 46 from phosphorylation 1094:1108 arg1 three 1121:1125 three 1121:1125 three 1121:1125 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 2 47 theme basic 339:343 arg1 PRP1 345:348 basic PRP1 339:348 basic PRP1 339:348 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 5 48 from one 796:798 arg1 bPRP3 803:807 bPRP3 803:807 bPRP3 803:807 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 49 theme other 978:982 arg1 pyro-Glu 1017:1024 N-terminal pyro-Glu 1006:1024 N-terminal pyro-Glu 1006:1024 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 49 theme other 978:982 arg1 modifications 984:996 possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications 680:996 possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu 680:1024 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 7 50 from bPRPs 1546:1550 arg1 patients 1580:1587 the Head and Neck Cancer patients 1555:1587 the Head and Neck Cancer patients 1555:1587 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 4 51 theme salivary 505:512 arg1 PRPs 514:517 salivary PRPs 505:517 salivary PRPs using LC-MALDI-TOF/TOF 505:540 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 5 52 gly N-glycosylation 728:742 arg2 sites 744:748 (i) N-glycosylation sites 724:748 the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3 701:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 53 from three 1121:1125 arg1 bPRP1 1130:1134 bPRP1 1130:1134 bPRP1 1130:1134 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 4 54 theme new 471:473 arg1 PTMs 475:478 new PTMs 471:478 new PTMs 471:478 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 5 55 theme peptidome 564:572 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 56 from one 813:815 arg1 bPRP4 820:824 bPRP4 820:824 bPRP4 820:824 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 57 theme allele 1194:1199 arg1 S 1201:1201 allele S 1194:1201 allele S 1194:1201 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 58 theme N-glycosylation 728:742 arg1 sites 744:748 (i) N-glycosylation sites 724:748 the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3 701:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 59 dep sites 848:852 arg1 ii 828:829 ii 828:829 ii 828:829 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 60 theme allele 1241:1246 arg1 bPRP4 1234:1238 bPRP4 1234:1238 bPRP4 (allele M) 1234:1249 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 60 theme allele 1241:1246 arg1 M 1248:1248 allele M 1241:1248 allele M 1241:1248 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 61 from two 855:857 arg1 one 872:874 one 872:874 one 872:874 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 61 from two 855:857 arg1 bPRP2 862:866 bPRP2 862:866 bPRP2 862:866 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 7 62 theme Neck 1568:1571 arg1 patients 1580:1587 the Head and Neck Cancer patients 1555:1587 the Head and Neck Cancer patients 1555:1587 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 6 63 theme data 1283:1286 arg1 analysis 1288:1295 salivary peptidome data analysis 1264:1295 salivary peptidome data analysis 1264:1295 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 5 64 from two 1027:1029 arg1 bPRP1 1034:1038 bPRP1 1034:1038 bPRP1 1034:1038 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 65 theme salivary 555:562 arg1 characterization 574:589 the salivary peptidome characterization 551:589 the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 551:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 66 theme variant 1217:1223 arg1 CP5 1225:1227 variant CP5 1217:1227 variant CP5 1217:1227 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 7 67 theme Head 1559:1562 arg1 patients 1580:1587 the Head and Neck Cancer patients 1555:1587 the Head and Neck Cancer patients 1555:1587 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 6 68 theme salivary 1264:1271 arg1 analysis 1288:1295 salivary peptidome data analysis 1264:1295 salivary peptidome data analysis 1264:1295 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 5 69 from six 1041:1043 arg1 bPRP2 1048:1052 bPRP2 1048:1052 bPRP2 1048:1052 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 70 theme different 597:605 arg1 subjects 607:614 20 different subjects 594:614 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups 594:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 2 71 theme acidic 298:303 arg1 PRP 305:307 acidic PRP 298:307 acidic PRP (aPRPs) 298:315 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 2 71 theme acidic 298:303 arg1 aPRPs 310:314 aPRPs 310:314 aPRPs 310:314 Up to date, only the serine phosphorylation at positions 8, 17, and 22 have been experimentally observed on acidic PRP (aPRPs), and at position 8 on basic PRP1 and 2. 20879038 5 72 from two 751:753 arg1 one 813:815 one 813:815 one 813:815 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 72 from two 751:753 arg1 protein 777:783 basic proline-rich protein 2 758:785 basic proline-rich protein 2 (bPRP2) 758:793 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 72 from two 751:753 arg1 bPRP2 788:792 bPRP2 788:792 bPRP2 788:792 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 72 from two 751:753 arg1 one 796:798 one 796:798 one 796:798 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 1 73 theme salivary 139:146 arg1 peptides 148:155 salivary peptides 139:155 salivary peptides 139:155 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 6 74 theme PRP-modified 1334:1345 arg1 residues 1347:1354 45 new PRP-modified residues 1327:1354 45 new PRP-modified residues 1327:1354 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 5 75 from eight 1055:1059 arg1 bPRP3 1064:1068 bPRP3 1064:1068 bPRP3 1064:1068 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 7 76 theme modification 1530:1541 arg1 predominance 1494:1505 a predominance 1492:1505 a predominance of N-acetyl hexosamine modification 1492:1541 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 5 77 theme proline-rich 764:775 arg1 one 813:815 one 813:815 one 813:815 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 77 theme proline-rich 764:775 arg1 protein 777:783 basic proline-rich protein 2 758:785 basic proline-rich protein 2 (bPRP2) 758:793 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 77 theme proline-rich 764:775 arg1 bPRP2 788:792 bPRP2 788:792 bPRP2 788:792 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 77 theme proline-rich 764:775 arg1 one 796:798 one 796:798 one 796:798 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 78 dep Control 621:627 arg1 groups 665:670 Cancer groups 658:670 Cancer groups 658:670 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 79 theme N-terminal 1006:1015 arg1 pyro-Glu 1017:1024 N-terminal pyro-Glu 1006:1024 N-terminal pyro-Glu 1006:1024 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 7 80 theme N-acetyl 1510:1517 arg1 modification 1530:1541 N-acetyl hexosamine modification 1510:1541 N-acetyl hexosamine modification 1510:1541 Moreover, comparing all subject groups, it was noticed a predominance of N-acetyl hexosamine modification on bPRPs in the Head and Neck Cancer patients. 20879038 5 81 dep phosphorylation 1094:1108 arg1 v 1091:1091 v 1091:1091 v 1091:1091 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 1 82 with family 129:134 arg1 PTMs 184:187 PTMs 184:187 PTMs 184:187 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 1 82 with family 129:134 arg1 isoforms 171:178 distinct isoforms 162:178 distinct isoforms 162:178 Proline-rich proteins (PRPs) are the most complex family of salivary peptides with distinct isoforms and PTMs. 20879038 0 83 theme new 8:10 arg1 modifications 30:42 new posttranslational modifications 8:42 new posttranslational modifications 8:42 Finding new posttranslational modifications in salivary proline-rich proteins. 20879038 5 84 theme terminal 897:904 arg1 sites 921:925 (iii) other terminal monosaccharide sites 885:925 (iii) other terminal monosaccharide sites 885:925 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 85 gly O-glycosylation 832:846 arg2 sites 848:852 (ii) O-glycosylation sites 827:852 (ii) O-glycosylation sites 827:852 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 6 86 theme peptidome 1273:1281 arg1 analysis 1288:1295 salivary peptidome data analysis 1264:1295 salivary peptidome data analysis 1264:1295 In summary, salivary peptidome data analysis allowed the identification of 45 new PRP-modified residues, mainly due to glycosylation, phosphorylation and conversion of Gln to pyro-Glu. 20879038 5 87 theme following 705:713 arg1 six 928:930 six 928:930 six 928:930 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 87 theme following 705:713 arg1 species 715:721 the following species 701:721 the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3 701:970 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 87 theme following 705:713 arg1 two 959:961 two 959:961 two 959:961 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 87 theme following 705:713 arg1 two 855:857 two 855:857 two 855:857 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 87 theme following 705:713 arg1 two 942:944 two 942:944 two 942:944 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 87 theme following 705:713 arg1 two 751:753 two 751:753 two 751:753 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 0 88 theme salivary 47:54 arg1 proteins 69:76 salivary proline-rich proteins 47:76 salivary proline-rich proteins 47:76 Finding new posttranslational modifications in salivary proline-rich proteins. 20879038 5 89 theme O-glycosylation 832:846 arg1 sites 848:852 (ii) O-glycosylation sites 827:852 (ii) O-glycosylation sites 827:852 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 3 90 theme glucoronyl 375:384 arg1 group 386:390 a glucoronyl group 373:390 a glucoronyl group 373:390 The presence of a glucoronyl group at Ser17 was also noticed on aPRP. 20879038 5 91 theme possible 680:687 arg1 pyro-Glu 1017:1024 N-terminal pyro-Glu 1006:1024 N-terminal pyro-Glu 1006:1024 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 91 theme possible 680:687 arg1 modifications 984:996 possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications 680:996 possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu 680:1024 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 92 from characterization 574:589 arg1 Head 644:647 Head 644:647 Head 644:647 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 92 from characterization 574:589 arg1 Control 621:627 Control 621:627 Control 621:627 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 92 from characterization 574:589 arg1 Neck 653:656 Neck 653:656 Neck 653:656 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 92 from characterization 574:589 arg1 Diabetic 630:637 Diabetic 630:637 Diabetic 630:637 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 93 dep sites 744:748 arg1 i 725:725 i 725:725 i 725:725 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 94 dep bPRP1 1187:1191 arg1 vi 1183:1184 vi 1183:1184 vi 1183:1184 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 94 dep bPRP1 1187:1191 arg1 S 1201:1201 allele S 1194:1201 allele S 1194:1201 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 94 dep bPRP1 1187:1191 arg1 M 1211:1211 allele M 1204:1211 allele M 1204:1211 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 5 94 dep bPRP1 1187:1191 arg1 CP5 1225:1227 variant CP5 1217:1227 variant CP5 1217:1227 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 20879038 4 95 theme PRPs 514:517 arg1 PTMs 475:478 new PTMs 471:478 new PTMs 471:478 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 4 95 theme PRPs 514:517 arg1 isoforms 493:500 distinct isoforms 484:500 distinct isoforms 484:500 The main goal of this study was to identify new PTMs and distinct isoforms of salivary PRPs using LC-MALDI-TOF/TOF. 20879038 5 96 theme allele 1204:1209 arg1 M 1211:1211 allele M 1204:1211 allele M 1204:1211 Through the salivary peptidome characterization of 20 different subjects from Control, Diabetic, and Head and Neck Cancer groups, it was possible to identify the following species: (i) N-glycosylation sites: two in basic proline-rich protein 2 (bPRP2), one in bPRP3 and one in bPRP4; (ii) O-glycosylation sites: two in bPRP2 and one in aPRP; (iii) other terminal monosaccharide sites: six in bPRP1, two in bPRP2 and two in bPRP3; (iv) other modifications such as N-terminal pyro-Glu (two in bPRP1, six in bPRP2, eight in bPRP3 and nine in bPRP4); (v) phosphorylation in serine, three in bPRP1, one in bPRP2, one in bPRP3 and one in aPRP1; (vi) bPRP1 (allele S, allele M and variant CP5) and bPRP4 (allele M). 9875849 1 0 theme IgE 146:148 arg1 receptor 150:157 the human high-affinity IgE receptor 122:157 the human high-affinity IgE receptor 122:157 Allergic responses result from the activation of mast cells by the human high-affinity IgE receptor. 9875849 4 1 theme bent 490:493 arg1 arrangement 495:505 a highly bent arrangement 481:505 a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE 481:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 6 2 theme therapeutic 837:847 arg1 approaches 849:858 new therapeutic approaches 833:858 new therapeutic approaches to allergy treatment 833:879 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 4 3 theme convex 555:560 arg1 surface 562:568 an extended convex surface 543:568 an extended convex surface of interaction with IgE 543:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 5 4 theme IgE 641:643 arg1 molecules 645:653 IgE molecules 641:653 IgE molecules 641:653 A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. 9875849 4 5 theme interaction 573:583 arg1 surface 562:568 an extended convex surface 543:568 an extended convex surface of interaction with IgE 543:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 1 6 theme Allergic 59:66 arg1 responses 68:76 Allergic responses 59:76 Allergic responses 59:76 Allergic responses result from the activation of mast cells by the human high-affinity IgE receptor. 9875849 2 7 theme response 269:276 arg1 initiation 253:262 the initiation 249:262 the initiation of a response 249:276 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 5 8 theme prominent 597:605 arg1 loop 607:610 A prominent loop 595:610 A prominent loop that confers specificity for IgE molecules 595:653 A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. 9875849 6 9 theme receptor 779:786 arg1 structure 758:766 The crystal structure 746:766 The crystal structure of the IgE receptor 746:786 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 3 10 theme human 419:423 arg1 receptor 429:436 the human IgE receptor 415:436 the human IgE receptor 415:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 5 11 theme exposed 725:731 arg1 tryptophans 733:743 four exposed tryptophans 720:743 four exposed tryptophans 720:743 A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. 9875849 3 12 theme antibody-binding 387:402 arg1 receptor 429:436 the human IgE receptor 415:436 the human IgE receptor 415:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 3 12 theme antibody-binding 387:402 arg1 domains 404:410 the antibody-binding domains 383:410 the antibody-binding domains of the human IgE receptor 383:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 6 13 theme IgE 775:777 arg1 receptor 779:786 the IgE receptor 771:786 the IgE receptor 771:786 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 3 14 theme IgE 425:427 arg1 receptor 429:436 the human IgE receptor 415:436 the human IgE receptor 415:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 3 15 theme crystal 362:368 arg1 structure 370:378 the X-ray crystal structure 352:378 the X-ray crystal structure of the antibody-binding domains of the human IgE receptor 352:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 6 16 theme allergy 863:869 arg1 treatment 871:879 allergy treatment 863:879 allergy treatment 863:879 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 2 17 theme IgE 302:304 arg1 binding 291:297 the binding 287:297 the binding of IgE to its high-affinity receptor 287:334 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 0 18 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human high-affinity IgE receptor 0:56 Crystal structure of the human high-affinity IgE receptor. 9875849 6 19 theme approaches 849:858 arg1 development 818:828 the development 814:828 the development of new therapeutic approaches to allergy treatment 814:879 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 3 20 theme X-ray 356:360 arg1 structure 370:378 the X-ray crystal structure 352:378 the X-ray crystal structure of the antibody-binding domains of the human IgE receptor 352:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 4 21 theme extended 546:553 arg1 surface 562:568 an extended convex surface 543:568 an extended convex surface of interaction with IgE 543:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 5 22 theme unusual 697:703 arg1 arrangement 705:715 an unusual arrangement 694:715 an unusual arrangement of four exposed tryptophans 694:743 A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. 9875849 3 23 theme A 445:445 arg1 resolution 447:456 2.4 A resolution 441:456 2.4 A resolution 441:456 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 0 24 theme high-affinity 31:43 arg1 receptor 49:56 the human high-affinity IgE receptor 21:56 the human high-affinity IgE receptor 21:56 Crystal structure of the human high-affinity IgE receptor. 9875849 2 25 theme high-affinity 313:325 arg1 receptor 327:334 its high-affinity receptor 309:334 its high-affinity receptor 309:334 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 2 26 theme allergic 173:180 arg1 reactions 182:190 IgE-mediated allergic reactions 160:190 IgE-mediated allergic reactions 160:190 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 2 27 theme compounds 234:242 arg1 variety 209:215 a variety 207:215 a variety of environmental compounds 207:242 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 2 27 theme compounds 234:242 arg1 compounds 234:242 environmental compounds 220:242 environmental compounds 220:242 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 3 28 theme receptor 429:436 arg1 receptor 429:436 the human IgE receptor 415:436 the human IgE receptor 415:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 3 28 theme receptor 429:436 arg1 domains 404:410 the antibody-binding domains 383:410 the antibody-binding domains of the human IgE receptor 383:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 0 29 theme human 25:29 arg1 receptor 49:56 the human high-affinity IgE receptor 21:56 the human high-affinity IgE receptor 21:56 Crystal structure of the human high-affinity IgE receptor. 9875849 3 30 theme domains 404:410 arg1 structure 370:378 the X-ray crystal structure 352:378 the X-ray crystal structure of the antibody-binding domains of the human IgE receptor 352:436 We have solved the X-ray crystal structure of the antibody-binding domains of the human IgE receptor at 2.4 A resolution. 9875849 2 31 theme IgE-mediated 160:171 arg1 reactions 182:190 IgE-mediated allergic reactions 160:190 IgE-mediated allergic reactions 160:190 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 2 32 theme environmental 220:232 arg1 compounds 234:242 environmental compounds 220:242 environmental compounds 220:242 IgE-mediated allergic reactions may develop to a variety of environmental compounds, but the initiation of a response requires the binding of IgE to its high-affinity receptor. 9875849 6 33 theme crystal 750:756 arg1 structure 758:766 The crystal structure 746:766 The crystal structure of the IgE receptor 746:786 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 0 34 theme receptor 49:56 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human high-affinity IgE receptor 0:56 Crystal structure of the human high-affinity IgE receptor. 9875849 6 35 theme new 833:835 arg1 approaches 849:858 new therapeutic approaches 833:858 new therapeutic approaches to allergy treatment 833:879 The crystal structure of the IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. 9875849 1 36 theme mast 108:111 arg1 cells 113:117 mast cells 108:117 mast cells 108:117 Allergic responses result from the activation of mast cells by the human high-affinity IgE receptor. 9875849 0 37 theme IgE 45:47 arg1 receptor 49:56 the human high-affinity IgE receptor 21:56 the human high-affinity IgE receptor 21:56 Crystal structure of the human high-affinity IgE receptor. 9875849 4 38 theme domains 525:531 arg1 arrangement 495:505 a highly bent arrangement 481:505 a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE 481:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 1 39 theme cells 113:117 arg1 activation 94:103 the activation 90:103 the activation of mast cells by the human high-affinity IgE receptor 90:157 Allergic responses result from the activation of mast cells by the human high-affinity IgE receptor. 9875849 4 40 theme immunoglobulin 510:523 arg1 domains 525:531 immunoglobulin domains 510:531 immunoglobulin domains that form an extended convex surface of interaction with IgE 510:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 4 41 with interaction 573:583 arg1 IgE 590:592 IgE 590:592 IgE 590:592 The structure reveals a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. 9875849 5 42 theme receptor 672:679 arg1 surface 681:687 the receptor surface 668:687 the receptor surface 668:687 A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. 9875849 5 43 theme tryptophans 733:743 arg1 arrangement 705:715 an unusual arrangement 694:715 an unusual arrangement of four exposed tryptophans 694:743 A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of four exposed tryptophans. 9875849 1 44 theme human 126:130 arg1 receptor 150:157 the human high-affinity IgE receptor 122:157 the human high-affinity IgE receptor 122:157 Allergic responses result from the activation of mast cells by the human high-affinity IgE receptor. 9875849 1 45 theme high-affinity 132:144 arg1 receptor 150:157 the human high-affinity IgE receptor 122:157 the human high-affinity IgE receptor 122:157 Allergic responses result from the activation of mast cells by the human high-affinity IgE receptor. 10206894 0 0 theme MHC 66:68 arg1 molecules 70:78 MHC molecules 66:78 MHC molecules 66:78 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 5 1 theme normal 692:697 arg1 conditions 715:724 normal or pathological conditions 692:724 normal or pathological conditions 692:724 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 3 2 theme class 441:445 arg1 chain 455:459 the class I light chain 437:459 the class I light chain beta2-microglobulin 437:479 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 5 3 theme lipid 669:673 arg1 catabolism 675:684 lipid catabolism 669:684 lipid catabolism 669:684 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 4 4 theme analogous 524:532 arg1 groove 517:522 a large groove 509:522 a large groove analogous to class I MHC peptide binding grooves 509:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 3 5 theme I 447:447 arg1 chain 455:459 the class I light chain 437:459 the class I light chain beta2-microglobulin 437:479 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 3 6 theme light 449:453 arg1 chain 455:459 the class I light chain 437:459 the class I light chain beta2-microglobulin 437:479 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 4 7 theme class 537:541 arg1 grooves 565:571 class I MHC peptide binding grooves 537:571 class I MHC peptide binding grooves 537:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 2 8 theme fat 251:253 arg1 losses 255:260 the extensive fat losses 237:260 the extensive fat losses associated with some advanced cancers 237:298 ZAG stimulates lipid degradation in adipocytes and causes the extensive fat losses associated with some advanced cancers. 10206894 3 9 theme angstrom 309:316 arg1 structure 326:334 The 2.8 angstrom crystal structure 301:334 The 2.8 angstrom crystal structure of ZAG 301:341 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 2 10 theme extensive 241:249 arg1 losses 255:260 the extensive fat losses 237:260 the extensive fat losses associated with some advanced cancers 237:298 ZAG stimulates lipid degradation in adipocytes and causes the extensive fat losses associated with some advanced cancers. 10206894 1 11 gly Zn-alpha2-glycoprotein 81:102 arg1 protein 123:129 a soluble protein 113:129 a soluble protein that is present in serum and other body fluids 113:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 11 gly Zn-alpha2-glycoprotein 81:102 arg1 ZAG 105:107 ZAG 105:107 ZAG 105:107 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 11 gly Zn-alpha2-glycoprotein 81:102 arg1 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein (ZAG) 81:108 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 11 gly Zn-alpha2-glycoprotein 81:102 arg1 present 139:145 present 139:145 present 139:145 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 5 12 theme pathological 702:713 arg1 conditions 715:724 normal or pathological conditions 692:724 normal or pathological conditions 692:724 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 3 13 theme class 355:359 arg1 MHC 397:399 MHC 397:399 MHC 397:399 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 3 13 theme class 355:359 arg1 complex 388:394 class I major histocompatibility complex 355:394 a class I major histocompatibility complex (MHC) heavy chain 353:412 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 2 14 from degradation 200:210 arg1 adipocytes 215:224 adipocytes 215:224 adipocytes 215:224 ZAG stimulates lipid degradation in adipocytes and causes the extensive fat losses associated with some advanced cancers. 10206894 3 15 theme heavy 402:406 arg1 chain 408:412 a class I major histocompatibility complex (MHC) heavy chain 353:412 a class I major histocompatibility complex (MHC) heavy chain 353:412 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 3 16 theme ZAG 339:341 arg1 structure 326:334 The 2.8 angstrom crystal structure 301:334 The 2.8 angstrom crystal structure of ZAG 301:341 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 2 17 theme advanced 283:290 arg1 cancers 292:298 some advanced cancers 278:298 some advanced cancers 278:298 ZAG stimulates lipid degradation in adipocytes and causes the extensive fat losses associated with some advanced cancers. 10206894 0 18 theme Crystal 0:6 arg1 factor 48:53 a fat-depleting factor 32:53 a fat-depleting factor related to MHC molecules 32:78 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 0 18 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human ZAG 0:29 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 1 19 located present 139:145 arg1 serum 150:154 serum 150:154 serum 150:154 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 19 located present 139:145 arg2 present 139:145 present 139:145 present 139:145 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 19 located present 139:145 arg2 protein 123:129 a soluble protein 113:129 a soluble protein that is present in serum and other body fluids 113:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 19 located present 139:145 arg2 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein (ZAG) 81:108 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 19 located present 139:145 arg1 fluids 171:176 other body fluids 160:176 other body fluids 160:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 0 20 theme human 21:25 arg1 ZAG 27:29 human ZAG 21:29 human ZAG 21:29 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 5 21 theme nonpeptidic 622:632 arg1 compound 634:641 a nonpeptidic compound 620:641 a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions 620:724 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 1 22 theme other 160:164 arg1 fluids 171:176 other body fluids 160:176 other body fluids 160:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 4 23 theme large 511:515 arg1 groove 517:522 a large groove 509:522 a large groove analogous to class I MHC peptide binding grooves 509:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 1 24 theme body 166:169 arg1 fluids 171:176 other body fluids 160:176 other body fluids 160:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 3 25 theme histocompatibility 369:386 arg1 MHC 397:399 MHC 397:399 MHC 397:399 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 3 25 theme histocompatibility 369:386 arg1 complex 388:394 class I major histocompatibility complex 355:394 a class I major histocompatibility complex (MHC) heavy chain 353:412 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 1 26 attach present 139:145 arg1 serum 150:154 serum 150:154 serum 150:154 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 26 attach present 139:145 arg2 present 139:145 present 139:145 present 139:145 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 26 attach present 139:145 arg2 protein 123:129 a soluble protein 113:129 a soluble protein that is present in serum and other body fluids 113:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 26 attach present 139:145 arg2 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein (ZAG) 81:108 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 26 attach present 139:145 arg1 fluids 171:176 other body fluids 160:176 other body fluids 160:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 0 27 theme ZAG 27:29 arg1 factor 48:53 a fat-depleting factor 32:53 a fat-depleting factor related to MHC molecules 32:78 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 0 27 theme ZAG 27:29 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human ZAG 0:29 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 3 28 theme complex 388:394 arg1 chain 408:412 a class I major histocompatibility complex (MHC) heavy chain 353:412 a class I major histocompatibility complex (MHC) heavy chain 353:412 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 2 29 theme lipid 194:198 arg1 degradation 200:210 lipid degradation 194:210 lipid degradation in adipocytes 194:224 ZAG stimulates lipid degradation in adipocytes and causes the extensive fat losses associated with some advanced cancers. 10206894 1 30 from fluids 171:176 arg1 protein 123:129 a soluble protein 113:129 a soluble protein that is present in serum and other body fluids 113:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 30 from fluids 171:176 arg1 present 139:145 present 139:145 present 139:145 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 30 from fluids 171:176 arg1 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein (ZAG) 81:108 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 3 31 theme crystal 318:324 arg1 structure 326:334 The 2.8 angstrom crystal structure 301:334 The 2.8 angstrom crystal structure of ZAG 301:341 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 0 32 theme fat-depleting 34:46 arg1 factor 48:53 a fat-depleting factor 32:53 a fat-depleting factor related to MHC molecules 32:78 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 0 32 theme fat-depleting 34:46 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human ZAG 0:29 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 5 33 contain contains 611:618 arg2 compound 634:641 a nonpeptidic compound 620:641 a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions 620:724 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 5 33 contain contains 611:618 arg1 groove 604:609 the ZAG groove 596:609 the ZAG groove 596:609 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 5 34 theme ZAG 600:602 arg1 groove 604:609 the ZAG groove 596:609 the ZAG groove 596:609 Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions. 10206894 3 35 theme major 363:367 arg1 MHC 397:399 MHC 397:399 MHC 397:399 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 3 35 theme major 363:367 arg1 complex 388:394 class I major histocompatibility complex 355:394 a class I major histocompatibility complex (MHC) heavy chain 353:412 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 4 36 theme ZAG 486:488 arg1 structure 490:498 The ZAG structure 482:498 The ZAG structure 482:498 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 4 37 theme MHC 545:547 arg1 grooves 565:571 class I MHC peptide binding grooves 537:571 class I MHC peptide binding grooves 537:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 1 38 from serum 150:154 arg1 protein 123:129 a soluble protein 113:129 a soluble protein that is present in serum and other body fluids 113:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 38 from serum 150:154 arg1 present 139:145 present 139:145 present 139:145 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 38 from serum 150:154 arg1 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein (ZAG) 81:108 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 4 39 theme I 543:543 arg1 grooves 565:571 class I MHC peptide binding grooves 537:571 class I MHC peptide binding grooves 537:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 0 40 theme related 55:61 arg1 factor 48:53 a fat-depleting factor 32:53 a fat-depleting factor related to MHC molecules 32:78 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 0 40 theme related 55:61 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human ZAG 0:29 Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. 10206894 1 41 theme soluble 115:121 arg1 protein 123:129 a soluble protein 113:129 a soluble protein that is present in serum and other body fluids 113:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 41 theme soluble 115:121 arg1 present 139:145 present 139:145 present 139:145 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 41 theme soluble 115:121 arg1 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein 81:102 Zn-alpha2-glycoprotein (ZAG) 81:108 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 4 42 theme binding 557:563 arg1 grooves 565:571 class I MHC peptide binding grooves 537:571 class I MHC peptide binding grooves 537:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 10206894 1 43 from present 139:145 arg1 serum 150:154 serum 150:154 serum 150:154 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 1 43 from present 139:145 arg1 fluids 171:176 other body fluids 160:176 other body fluids 160:176 Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. 10206894 3 44 theme chain 455:459 arg1 beta2-microglobulin 461:479 the class I light chain beta2-microglobulin 437:479 the class I light chain beta2-microglobulin 437:479 The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. 10206894 4 45 theme peptide 549:555 arg1 grooves 565:571 class I MHC peptide binding grooves 537:571 class I MHC peptide binding grooves 537:571 The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. 2226832 5 0 theme human 613:617 arg1 elastase 630:637 human neutrophil elastase 613:637 human neutrophil elastase 613:637 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 1 1 theme antibacterial 255:267 arg1 activity 303:310 antibacterial and monocyte-specific chemotactic activity 255:310 antibacterial and monocyte-specific chemotactic activity 255:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 0 2 theme antibacterial 65:77 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 2 theme antibacterial 65:77 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 5 3 theme neutrophil 619:628 arg1 elastase 630:637 human neutrophil elastase 613:637 human neutrophil elastase 613:637 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 6 4 theme active 778:783 arg1 serine 799:804 serine 799:804 serine 799:804 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 4 theme active 778:783 arg1 residues 790:797 the active site residues 774:797 the active site residues serine and histidine 774:818 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 4 theme active 778:783 arg1 histidine 810:818 histidine 810:818 histidine 810:818 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 0 5 theme chemotactic 101:111 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 5 theme chemotactic 101:111 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 1 6 theme monocyte-specific 273:289 arg1 activity 303:310 antibacterial and monocyte-specific chemotactic activity 255:310 antibacterial and monocyte-specific chemotactic activity 255:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 0 7 theme monocyte-specific 83:99 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 7 theme monocyte-specific 83:99 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 5 8 theme several 663:669 arg1 proteinases 692:702 several other granule serine proteinases 663:702 several other granule serine proteinases 663:702 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 1 9 theme chemotactic 291:301 arg1 activity 303:310 antibacterial and monocyte-specific chemotactic activity 255:310 antibacterial and monocyte-specific chemotactic activity 255:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 4 10 from Asn-110 558:564 arg1 others 536:541 others 536:541 others at Asn-114 and Asn-110 536:564 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 4 11 theme CAP37 470:474 arg1 species 459:465 Some species 454:465 Some species of CAP37 454:474 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 5 12 theme % 590:590 arg1 identity 601:608 45% sequence identity 588:608 45% sequence identity to human neutrophil elastase 588:637 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 5 13 theme 45 588:589 arg1 % 590:590 % 590:590 % 590:590 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 1 14 theme amino 185:189 arg1 sequence 196:203 the amino acid sequence 181:203 the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity 181:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 5 15 theme sequence 592:599 arg1 identity 601:608 45% sequence identity 588:608 45% sequence identity to human neutrophil elastase 588:637 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 6 16 theme serine 748:753 arg1 proteinase 755:764 a serine proteinase 746:764 a serine proteinase 746:764 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 16 theme serine 748:753 arg1 CAP37 733:737 CAP37 733:737 CAP37 733:737 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 1 17 theme acid 191:194 arg1 sequence 196:203 the amino acid sequence 181:203 the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity 181:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 0 18 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 18 theme acid 6:9 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 3 19 gly N-glycosylation 397:411 arg2 three 391:395 three 391:395 three 391:395 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 3 19 gly N-glycosylation 397:411 arg2 sites 413:417 three N-glycosylation sites 391:417 three N-glycosylation sites 391:417 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 3 20 contain has 387:389 arg1 It 384:385 It 384:385 It 384:385 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 3 20 contain has 387:389 arg2 sites 413:417 three N-glycosylation sites 391:417 three N-glycosylation sites 391:417 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 0 21 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 21 theme Amino 0:4 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 6 22 dep residues 790:797 arg1 serine 799:804 serine 799:804 serine 799:804 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 22 dep residues 790:797 arg1 residues 790:797 the active site residues 774:797 the active site residues serine and histidine 774:818 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 22 dep residues 790:797 arg1 histidine 810:818 histidine 810:818 histidine 810:818 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 5 23 theme serine 685:690 arg1 proteinases 692:702 several other granule serine proteinases 663:702 several other granule serine proteinases 663:702 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 0 24 gly glycoprotein 113:124 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 24 gly glycoprotein 113:124 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 1 25 theme CAP37 208:212 arg1 sequence 196:203 the amino acid sequence 181:203 the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity 181:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 3 26 theme N-glycosylation 397:411 arg1 sites 413:417 three N-glycosylation sites 391:417 three N-glycosylation sites 391:417 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 0 27 theme similar 139:145 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 27 theme similar 139:145 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 4 28 gly glycosylated 480:491 arg2 sites 506:510 all three sites 496:510 all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145 496:575 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 4 28 gly glycosylated 480:491 arg1 species 459:465 Some species 454:465 Some species of CAP37 454:474 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 5 29 theme other 671:675 arg1 proteinases 692:702 several other granule serine proteinases 663:702 several other granule serine proteinases 663:702 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 5 30 theme granule 677:683 arg1 proteinases 692:702 several other granule serine proteinases 663:702 several other granule serine proteinases 663:702 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 0 31 theme CAP37 23:27 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 31 theme CAP37 23:27 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 32 theme neutrophil 150:159 arg1 elastase 161:168 neutrophil elastase 150:168 neutrophil elastase 150:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 1 33 theme human 217:221 arg1 protein 242:248 a human neutrophil granule protein 215:248 a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity 215:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 1 33 theme human 217:221 arg1 CAP37 208:212 CAP37 208:212 CAP37 208:212 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 0 34 theme human 32:36 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 34 theme human 32:36 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 4 35 dep sites 506:510 arg1 others 536:541 others 536:541 others at Asn-114 and Asn-110 536:564 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 4 35 dep sites 506:510 arg1 some 513:516 some 513:516 some 513:516 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 4 35 dep sites 506:510 arg1 Asn-145 569:575 Asn-145 569:575 Asn-145 569:575 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 2226832 2 36 theme acid 369:372 arg1 residues 374:381 222 amino acid residues 359:381 222 amino acid residues 359:381 CAP37 is a single-chain protein consisting of 222 amino acid residues. 2226832 3 37 theme Asn 423:425 arg1 145 449:451 145 449:451 145 449:451 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 3 37 theme Asn 423:425 arg1 residues 427:434 Asn residues 100 423:438 Asn residues 100 423:438 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 3 37 theme Asn 423:425 arg1 114 441:443 114 441:443 114 441:443 It has three N-glycosylation sites, at Asn residues 100, 114 and 145. 2226832 5 38 theme 30-37 644:648 arg1 % 649:649 % 649:649 % 649:649 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 0 39 link granule-derived 49:63 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 39 link granule-derived 49:63 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 2 40 theme amino 363:367 arg1 residues 374:381 222 amino acid residues 359:381 222 amino acid residues 359:381 CAP37 is a single-chain protein consisting of 222 amino acid residues. 2226832 5 41 theme % 649:649 arg1 identity 651:658 30-37% identity 644:658 30-37% identity to several other granule serine proteinases 644:702 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 2 42 theme single-chain 324:335 arg1 CAP37 313:317 CAP37 313:317 CAP37 313:317 CAP37 is a single-chain protein consisting of 222 amino acid residues. 2226832 2 42 theme single-chain 324:335 arg1 protein 337:343 a single-chain protein 322:343 a single-chain protein consisting of 222 amino acid residues 322:381 CAP37 is a single-chain protein consisting of 222 amino acid residues. 2226832 1 43 theme neutrophil 223:232 arg1 protein 242:248 a human neutrophil granule protein 215:248 a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity 215:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 1 43 theme neutrophil 223:232 arg1 CAP37 208:212 CAP37 208:212 CAP37 208:212 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 1 44 with protein 242:248 arg1 activity 303:310 antibacterial and monocyte-specific chemotactic activity 255:310 antibacterial and monocyte-specific chemotactic activity 255:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 1 45 theme granule 234:240 arg1 protein 242:248 a human neutrophil granule protein 215:248 a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity 215:310 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 1 45 theme granule 234:240 arg1 CAP37 208:212 CAP37 208:212 CAP37 208:212 We report the amino acid sequence of CAP37, a human neutrophil granule protein with antibacterial and monocyte-specific chemotactic activity. 2226832 0 46 theme granule-derived 49:63 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 46 theme granule-derived 49:63 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 6 47 theme site 785:788 arg1 serine 799:804 serine 799:804 serine 799:804 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 47 theme site 785:788 arg1 residues 790:797 the active site residues 774:797 the active site residues serine and histidine 774:818 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 6 47 theme site 785:788 arg1 histidine 810:818 histidine 810:818 histidine 810:818 Despite these similarities, CAP37 is not a serine proteinase because the active site residues serine and histidine are replaced. 2226832 5 48 contain has 584:586 arg1 CAP37 578:582 CAP37 578:582 CAP37 578:582 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 5 48 contain has 584:586 arg2 identity 651:658 30-37% identity 644:658 30-37% identity to several other granule serine proteinases 644:702 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 5 48 contain has 584:586 arg2 identity 601:608 45% sequence identity 588:608 45% sequence identity to human neutrophil elastase 588:637 CAP37 has 45% sequence identity to human neutrophil elastase, and 30-37% identity to several other granule serine proteinases. 2226832 0 49 theme neutrophil 38:47 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of CAP37 0:27 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 0 49 theme neutrophil 38:47 arg1 glycoprotein 113:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein 30:124 a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase 30:168 Amino acid sequence of CAP37, a human neutrophil granule-derived antibacterial and monocyte-specific chemotactic glycoprotein structurally similar to neutrophil elastase. 2226832 4 50 from Asn-114 546:552 arg1 others 536:541 others 536:541 others at Asn-114 and Asn-110 536:564 Some species of CAP37 are glycosylated at all three sites; some at Asn-114 alone, others at Asn-114 and Asn-110 or Asn-145. 8393577 2 0 theme lysosomal 455:463 arg1 targeting 465:473 lysosomal targeting 455:473 lysosomal targeting 455:473 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 3 1 theme inhibitor-enzyme 698:713 arg1 interactions 715:726 inhibitor-enzyme interactions 698:726 inhibitor-enzyme interactions 698:726 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 2 2 theme crystal 327:333 arg1 structures 335:344 the crystal structures 323:344 the crystal structures of cathepsin D 323:359 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 1 3 theme protein 212:218 arg1 catabolism 220:229 protein catabolism 212:229 protein catabolism 212:229 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 0 4 theme D 68:68 arg1 forms 43:47 native and inhibited forms 22:47 native and inhibited forms of human cathepsin D 22:68 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 3 5 theme subsite 675:681 arg1 structures 683:692 subsite structures 675:692 subsite structures 675:692 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 3 6 theme D 879:879 arg1 inhibitors 881:890 cathepsin D inhibitors 869:890 cathepsin D inhibitors 869:890 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 2 7 from resolution 399:408 arg1 Determination 306:318 Determination 306:318 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution 306:408 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 4 8 theme phosphotransferase 931:948 arg1 region 962:967 a phosphotransferase recognition region 929:967 a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70 929:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 4 8 theme phosphotransferase 931:948 arg1 distant 1034:1040 distant 1034:1040 distant 1034:1040 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 5 9 theme structural 1334:1343 arg1 explanation 1345:1355 a structural explanation 1332:1355 a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface 1332:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 4 10 from site 1074:1077 arg1 region 962:967 a phosphotransferase recognition region 929:967 a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70 929:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 4 10 from site 1074:1077 arg1 distant 1034:1040 distant 1034:1040 distant 1034:1040 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 5 11 theme D 1146:1146 arg1 structure 1123:1131 the crystal structure 1111:1131 the crystal structure of cathepsin D 1111:1146 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 4 12 theme glycosylation 1060:1072 arg1 site 1074:1077 the N-domain glycosylation site 1047:1077 the N-domain glycosylation site at Asn-70 1047:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 1 13 theme antigen 232:238 arg1 processing 240:249 antigen processing 232:249 antigen processing 232:249 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 2 14 theme two-chain 484:492 arg1 protease 519:526 this two-chain, N-glycosylated aspartic protease 479:526 this two-chain, N-glycosylated aspartic protease 479:526 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 2 15 with complex 367:373 arg1 pepstatin 380:388 pepstatin 380:388 pepstatin 380:388 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 5 16 theme phosphotransferase 1275:1292 arg1 region 1306:1311 the phosphotransferase recognition region 1271:1311 the phosphotransferase recognition region 1271:1311 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 5 17 link N-linked 1177:1184 arg1 oligosaccharide 1186:1200 an N-linked oligosaccharide 1174:1200 an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface 1174:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 0 18 theme lysosomal 88:96 arg1 targeting 98:106 lysosomal targeting 88:106 lysosomal targeting 88:106 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 3 19 theme cathepsin 869:877 arg1 inhibitors 881:890 cathepsin D inhibitors 869:890 cathepsin D inhibitors 869:890 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 5 20 theme region 1306:1311 arg1 Lys-203 1234:1240 Lys-203 1234:1240 Lys-203 1234:1240 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 5 20 theme region 1306:1311 arg1 component 1258:1266 a key component 1252:1266 a key component of the phosphotransferase recognition region 1252:1311 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 3 21 theme structure-activity 778:795 arg1 relationships 797:809 structure-activity relationships 778:809 structure-activity relationships 778:809 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 5 22 theme N-linked 1177:1184 arg1 oligosaccharide 1186:1200 an N-linked oligosaccharide 1174:1200 an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface 1174:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 3 23 from differences 660:670 arg1 structures 683:692 subsite structures 675:692 subsite structures 675:692 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 3 23 from differences 660:670 arg1 interactions 715:726 inhibitor-enzyme interactions 698:726 inhibitor-enzyme interactions 698:726 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 4 24 theme recognition 950:960 arg1 region 962:967 a phosphotransferase recognition region 929:967 a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70 929:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 4 24 theme recognition 950:960 arg1 distant 1034:1040 distant 1034:1040 distant 1034:1040 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 2 25 theme D 359:359 arg1 structures 335:344 the crystal structures 323:344 the crystal structures of cathepsin D 323:359 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 2 25 theme D 359:359 arg1 complex 367:373 a complex 365:373 a complex with pepstatin 365:388 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 0 26 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 0:123 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 4 27 theme Mutagenesis 893:903 arg1 studies 905:911 Mutagenesis studies 893:911 Mutagenesis studies 893:911 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 2 28 theme cathepsin 349:357 arg1 D 359:359 cathepsin D 349:359 cathepsin D 349:359 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 0 29 theme native 22:27 arg1 forms 43:47 native and inhibited forms 22:47 native and inhibited forms of human cathepsin D 22:68 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 0 30 theme drug 112:115 arg1 design 117:122 drug design 112:122 drug design 112:122 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 1 31 theme lysosomal 156:164 arg1 protease 166:173 a lysosomal protease 154:173 a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression 154:303 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 1 31 theme lysosomal 156:164 arg1 D 135:135 Cathepsin D 125:135 Cathepsin D (EC 3.4.23.5) 125:149 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 1 32 theme degenerative 252:263 arg1 diseases 265:272 degenerative diseases 252:272 degenerative diseases 252:272 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 3 33 theme human 621:625 arg1 complex 643:649 a human renin-inhibitor complex 619:649 a human renin-inhibitor complex 619:649 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 0 34 theme inhibited 33:41 arg1 forms 43:47 native and inhibited forms 22:47 native and inhibited forms of human cathepsin D 22:68 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 3 35 theme affinity 753:760 arg1 differences 762:772 affinity differences 753:772 affinity differences 753:772 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 5 36 theme oligosaccharide 1186:1200 arg1 presence 1162:1169 the presence 1158:1169 the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface 1158:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 3 37 theme renin-inhibitor 627:641 arg1 complex 643:649 a human renin-inhibitor complex 619:649 a human renin-inhibitor complex 619:649 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 4 38 theme oligosaccharide 990:1004 arg1 phosphorylation 1006:1020 oligosaccharide phosphorylation 990:1020 oligosaccharide phosphorylation 990:1020 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 2 39 theme inhibitor 433:441 arg1 binding 443:449 inhibitor binding 433:449 inhibitor binding 433:449 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 4 40 from Asn-70 1082:1087 arg1 site 1074:1077 the N-domain glycosylation site 1047:1077 the N-domain glycosylation site at Asn-70 1047:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 2 41 theme aspartic 510:517 arg1 protease 519:526 this two-chain, N-glycosylated aspartic protease 479:526 this two-chain, N-glycosylated aspartic protease 479:526 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 3 42 theme pepstatin 576:584 arg1 complex 565:571 a complex 563:571 a complex of pepstatin bound to rhizopuspepsin 563:608 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 5 43 theme key 1254:1256 arg1 Lys-203 1234:1240 Lys-203 1234:1240 Lys-203 1234:1240 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 5 43 theme key 1254:1256 arg1 component 1258:1266 a key component 1252:1266 a key component of the phosphotransferase recognition region 1252:1311 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 2 44 theme complex 367:373 arg1 Determination 306:318 Determination 306:318 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution 306:408 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 5 45 theme cathepsin 1136:1144 arg1 D 1146:1146 cathepsin D 1136:1146 cathepsin D 1136:1146 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 5 46 theme crystal 1115:1121 arg1 structure 1123:1131 the crystal structure 1111:1131 the crystal structure of cathepsin D 1111:1146 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 0 47 theme forms 43:47 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 0:123 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 1 48 theme breast 279:284 arg1 progression 293:303 breast cancer progression 279:303 breast cancer progression 279:303 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 2 49 dep two-chain 484:492 arg1 N-glycosylated 495:508 N-glycosylated 495:508 N-glycosylated 495:508 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 5 50 theme distant 1413:1419 arg1 sites 1421:1425 apparently distant sites 1402:1425 apparently distant sites on the protein surface 1402:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 3 51 theme complex 565:571 arg1 structures 549:558 the structures 545:558 the structures of a complex of pepstatin bound to rhizopuspepsin 545:608 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 1 52 theme cancer 286:291 arg1 progression 293:303 breast cancer progression 279:303 breast cancer progression 279:303 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 4 53 gly glycosylation 1060:1072 arg2 Asn-70 1082:1087 Asn-70 1082:1087 Asn-70 1082:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 4 53 gly glycosylation 1060:1072 arg2 site 1074:1077 the N-domain glycosylation site 1047:1077 the N-domain glycosylation site at Asn-70 1047:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 3 54 with Comparison 529:538 arg1 structures 549:558 the structures 545:558 the structures of a complex of pepstatin bound to rhizopuspepsin 545:608 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 3 54 with Comparison 529:538 arg1 complex 643:649 a human renin-inhibitor complex 619:649 a human renin-inhibitor complex 619:649 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 5 55 theme Electron 1090:1097 arg1 density 1099:1105 Electron density 1090:1105 Electron density for the crystal structure of cathepsin D 1090:1146 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 5 56 theme recognition 1294:1304 arg1 region 1306:1311 the phosphotransferase recognition region 1271:1311 the phosphotransferase recognition region 1271:1311 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 5 57 theme protein 1434:1440 arg1 surface 1442:1448 the protein surface 1430:1448 the protein surface 1430:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 3 58 theme inhibitors 881:890 arg1 specificity 854:864 the specificity 850:864 the specificity of cathepsin D inhibitors 850:890 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 1 59 theme important 193:201 arg1 roles 203:207 important roles 193:207 important roles 193:207 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 0 60 theme cathepsin 58:66 arg1 D 68:68 human cathepsin D 52:68 human cathepsin D 52:68 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 2 61 with structures 335:344 arg1 pepstatin 380:388 pepstatin 380:388 pepstatin 380:388 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 8393577 0 62 dep structures 8:17 arg1 implications 71:82 implications 71:82 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 0:123 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 1 63 theme Cathepsin 125:133 arg1 protease 166:173 a lysosomal protease 154:173 a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression 154:303 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 1 63 theme Cathepsin 125:133 arg1 D 135:135 Cathepsin D 125:135 Cathepsin D (EC 3.4.23.5) 125:149 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 1 63 theme Cathepsin 125:133 arg1 EC 138:139 EC 3.4.23.5 138:148 EC 3.4.23.5 138:148 Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. 8393577 3 64 with consistent 737:746 arg1 differences 762:772 affinity differences 753:772 affinity differences 753:772 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 3 64 with consistent 737:746 arg1 relationships 797:809 structure-activity relationships 778:809 structure-activity relationships 778:809 Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. 8393577 0 65 theme human 52:56 arg1 D 68:68 human cathepsin D 52:68 human cathepsin D 52:68 Crystal structures of native and inhibited forms of human cathepsin D: implications for lysosomal targeting and drug design. 8393577 4 66 theme N-domain 1051:1058 arg1 site 1074:1077 the N-domain glycosylation site 1047:1077 the N-domain glycosylation site at Asn-70 1047:1087 Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 A distant from the N-domain glycosylation site at Asn-70. 8393577 5 67 from sites 1421:1425 arg1 surface 1442:1448 the protein surface 1430:1448 the protein surface 1430:1448 Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface. 8393577 2 68 theme structures 335:344 arg1 Determination 306:318 Determination 306:318 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution 306:408 Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 A resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. 15628971 7 0 gly microheterogeneity 1564:1581 arg1 oligosaccharides 1595:1610 N-linked oligosaccharides 1586:1610 N-linked oligosaccharides disappeared 1586:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 1 theme form 656:659 arg1 production 631:640 (ii) production 626:640 (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 626:716 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 2 theme C4ST-1 664:669 arg1 form 656:659 the active form 645:659 the active form of C4ST-1 645:669 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 2 3 theme chondrosarcoma 308:321 arg1 cells 323:327 rat chondrosarcoma cells 304:327 rat chondrosarcoma cells 304:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 0 4 link N-linked 0:7 arg1 oligosaccharides 9:24 N-linked oligosaccharides 0:24 N-linked oligosaccharides 0:24 N-linked oligosaccharides are required to produce and stabilize the active form of chondroitin 4-sulphotransferase-1. 15628971 0 5 theme N-linked 0:7 arg1 oligosaccharides 9:24 N-linked oligosaccharides 0:24 N-linked oligosaccharides 0:24 N-linked oligosaccharides are required to produce and stabilize the active form of chondroitin 4-sulphotransferase-1. 15628971 5 6 attach attachment 859:868 arg3 region 909:914 the C-terminal region 894:914 the C-terminal region 894:914 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 6 attach attachment 859:868 arg2 N-glycan 882:889 a single N-glycan 873:889 a single N-glycan 873:889 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 7 theme C-terminal 1758:1767 arg1 region 1769:1774 the C-terminal region 1754:1774 the C-terminal region 1754:1774 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 8 theme COS-7 674:678 arg1 cells 680:684 COS-7 cells 674:684 COS-7 cells transfected with cDNA of C4ST-1 674:716 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 9 theme oligosaccharides 1595:1610 arg1 microheterogeneity 1564:1581 microheterogeneity 1564:1581 microheterogeneity of N-linked oligosaccharides disappeared 1564:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 1 10 theme chondroitin 126:136 arg1 4-sulphotransferase-1 138:158 chondroitin 4-sulphotransferase-1 126:158 chondroitin 4-sulphotransferase-1 126:158 C4ST-1 (chondroitin 4-sulphotransferase-1) transfers sulphate to position 4 of N-acetylgalactosamine in chondroitin. 15628971 1 10 theme chondroitin 126:136 arg1 C4ST-1 118:123 C4ST-1 118:123 C4ST-1 (chondroitin 4-sulphotransferase-1) 118:159 C4ST-1 (chondroitin 4-sulphotransferase-1) transfers sulphate to position 4 of N-acetylgalactosamine in chondroitin. 15628971 4 11 theme oligosaccharides 472:487 arg1 role 451:454 the functional role 436:454 the functional role of the N-linked oligosaccharides attached to C4ST-1 436:506 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 5 12 theme C4ST-1 711:716 arg1 cDNA 703:706 cDNA 703:706 cDNA of C4ST-1 703:716 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 13 link N-linked 1682:1689 arg1 oligosaccharides 1691:1706 N-linked oligosaccharides 1682:1706 N-linked oligosaccharides 1682:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 2 14 theme purified 261:268 arg1 C4ST-1 270:275 purified C4ST-1 261:275 purified C4ST-1 from the culture medium of rat chondrosarcoma cells 261:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 2 14 theme purified 261:268 arg1 glycoprotein 335:346 a glycoprotein 333:346 a glycoprotein containing approx 333:364 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 5 15 theme peptide 564:570 arg1 F 586:586 peptide N-glycosidase F 564:586 peptide N-glycosidase F 564:586 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 16 theme C4ST-1 827:832 arg1 region 817:822 the C-terminal region 802:822 the C-terminal region of C4ST-1 802:832 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 17 dep treatment 527:535 arg1 i 524:524 i 524:524 i 524:524 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 18 from region 1170:1175 arg1 site 1147:1150 the N-glycosylation site 1127:1150 the N-glycosylation site at the C-terminal region resulted in total loss of activity 1127:1210 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 19 theme single 875:880 arg1 N-glycan 882:889 a single N-glycan 873:889 a single N-glycan 873:889 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 20 theme protein 1541:1547 arg1 band 1520:1523 a broad protein band 1504:1523 a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared 1504:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 20 theme protein 1541:1547 arg1 bands 1650:1654 four discrete protein bands 1628:1654 four discrete protein bands with different numbers of N-linked oligosaccharides 1628:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 21 with bands 1650:1654 arg1 numbers 1671:1677 different numbers 1661:1677 different numbers of N-linked oligosaccharides 1661:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 22 theme activity 1203:1210 arg1 loss 1195:1198 total loss 1189:1198 total loss of activity 1189:1210 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 2 23 theme culture 286:292 arg1 medium 294:299 the culture medium 282:299 the culture medium of rat chondrosarcoma cells 282:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 3 24 theme 35 367:368 arg1 % 369:369 % 369:369 % 369:369 35% N-linked oligosaccharides. 15628971 5 25 theme recombinant 1060:1070 arg1 protein 1072:1078 the control recombinant protein 1048:1078 the control recombinant protein 1048:1078 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 26 theme C4ST-1 552:557 arg1 treatment 527:535 (i) treatment 523:535 (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F 523:586 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 27 theme N-linked 1682:1689 arg1 oligosaccharides 1691:1706 N-linked oligosaccharides 1682:1706 N-linked oligosaccharides 1682:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 28 theme broad 1506:1510 arg1 band 1520:1523 a broad protein band 1504:1523 a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared 1504:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 0 29 theme chondroitin 83:93 arg1 4-sulphotransferase-1 95:115 chondroitin 4-sulphotransferase-1 83:115 chondroitin 4-sulphotransferase-1 83:115 N-linked oligosaccharides are required to produce and stabilize the active form of chondroitin 4-sulphotransferase-1. 15628971 3 30 theme N-linked 371:378 arg1 oligosaccharides 380:395 35% N-linked oligosaccharides 367:395 35% N-linked oligosaccharides. 367:396 35% N-linked oligosaccharides. 15628971 5 31 dep truncation 1089:1098 arg1 v 1086:1086 v 1086:1086 v 1086:1086 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 6 32 dep production 1317:1326 arg1 the 1313:1315 the 1313:1315 the 1313:1315 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 5 33 theme active 944:949 arg1 form 951:954 the active form 940:954 the active form of C4ST-1 940:964 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 34 theme protein 1642:1648 arg1 bands 1650:1654 four discrete protein bands 1628:1654 four discrete protein bands with different numbers of N-linked oligosaccharides 1628:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 35 dep attachment 859:868 arg1 iv 855:856 iv 855:856 iv 855:856 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 36 theme located 791:797 arg1 site 786:789 the N-glycosylation site 766:789 the N-glycosylation site located at the C-terminal region of C4ST-1 766:832 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 37 theme C-terminal 898:907 arg1 region 909:914 the C-terminal region 894:914 the C-terminal region 894:914 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 38 gly N-glycosylation 1131:1145 arg2 region 1170:1175 the C-terminal region 1155:1175 the C-terminal region 1155:1175 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 38 gly N-glycosylation 1131:1145 arg2 site 1147:1150 the N-glycosylation site 1127:1150 the N-glycosylation site at the C-terminal region resulted in total loss of activity 1127:1210 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 6 39 theme C4ST-1 1364:1369 arg1 form 1356:1359 the active form 1345:1359 the active form of C4ST-1 1345:1369 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 5 40 theme region 1114:1119 arg1 activity 844:851 activity 844:851 activity 844:851 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 40 theme region 1114:1119 arg1 truncation 1089:1098 (v) truncation 1085:1098 (v) truncation of C-terminal region 1085:1119 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 2 41 from medium 294:299 arg1 C4ST-1 270:275 purified C4ST-1 261:275 purified C4ST-1 from the culture medium of rat chondrosarcoma cells 261:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 2 41 from medium 294:299 arg1 glycoprotein 335:346 a glycoprotein 333:346 a glycoprotein containing approx 333:364 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 6 42 theme form 1356:1359 arg1 stability 1332:1340 stability 1332:1340 stability 1332:1340 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 6 42 theme form 1356:1359 arg1 production 1317:1326 production 1317:1326 production 1317:1326 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 7 43 theme glycosylation 1461:1473 arg1 pattern 1475:1481 the glycosylation pattern 1457:1481 the glycosylation pattern of recombinant C4ST 1457:1501 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 44 dep abolished 834:842 arg1 deletion 754:761 deletion 754:761 deletion 754:761 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 44 dep abolished 834:842 arg1 iii 749:751 iii 749:751 iii 749:751 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 45 theme N-glycosylation 770:784 arg1 site 786:789 the N-glycosylation site 766:789 the N-glycosylation site located at the C-terminal region of C4ST-1 766:832 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 46 theme recombinant 985:995 arg1 enzyme 997:1002 the resulting recombinant enzyme 971:1002 the resulting recombinant enzyme 971:1002 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 46 theme recombinant 985:995 arg1 unstable 1018:1025 unstable 1018:1025 unstable 1018:1025 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 2 47 gly glycoprotein 335:346 arg1 C4ST-1 270:275 purified C4ST-1 261:275 purified C4ST-1 from the culture medium of rat chondrosarcoma cells 261:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 2 47 gly glycoprotein 335:346 arg1 glycoprotein 335:346 a glycoprotein 333:346 a glycoprotein containing approx 333:364 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 5 48 gly N-glycosylation 770:784 arg2 site 786:789 the N-glycosylation site 766:789 the N-glycosylation site located at the C-terminal region of C4ST-1 766:832 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 49 theme C-terminal 1159:1168 arg1 region 1170:1175 the C-terminal region 1155:1175 the C-terminal region 1155:1175 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 50 theme C-terminal 1421:1430 arg1 region 1432:1437 the C-terminal region 1417:1437 the C-terminal region 1417:1437 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 6 51 attach attached 1280:1287 arg2 oligosaccharides 1263:1278 N-linked oligosaccharides 1254:1278 N-linked oligosaccharides attached to C4ST-1 1254:1297 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 6 51 attach attached 1280:1287 arg1 C4ST-1 1292:1297 C4ST-1 1292:1297 C4ST-1 1292:1297 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 5 52 theme active 649:654 arg1 form 656:659 the active form 645:659 the active form of C4ST-1 645:669 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 3 53 link N-linked 371:378 arg1 oligosaccharides 380:395 35% N-linked oligosaccharides 367:395 35% N-linked oligosaccharides. 367:396 35% N-linked oligosaccharides. 15628971 5 54 theme N-glycosylation 1131:1145 arg1 site 1147:1150 the N-glycosylation site 1127:1150 the N-glycosylation site at the C-terminal region resulted in total loss of activity 1127:1210 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 55 theme N-linked 1389:1396 arg1 oligosaccharide 1398:1412 the N-linked oligosaccharide 1385:1412 the N-linked oligosaccharide at the C-terminal region 1385:1437 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 4 56 theme functional 440:449 arg1 role 451:454 the functional role 436:454 the functional role of the N-linked oligosaccharides attached to C4ST-1 436:506 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 2 57 theme rat 304:306 arg1 cells 323:327 rat chondrosarcoma cells 304:327 rat chondrosarcoma cells 304:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 4 58 theme present 405:411 arg1 paper 413:417 the present paper 401:417 the present paper 401:417 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 7 59 link N-linked 1389:1396 arg1 oligosaccharide 1398:1412 the N-linked oligosaccharide 1385:1412 the N-linked oligosaccharide at the C-terminal region 1385:1437 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 1 60 theme N-acetylgalactosamine 197:217 arg1 position 183:190 position 4 183:192 position 4 of N-acetylgalactosamine in chondroitin 183:232 C4ST-1 (chondroitin 4-sulphotransferase-1) transfers sulphate to position 4 of N-acetylgalactosamine in chondroitin. 15628971 7 61 theme N-linked 1586:1593 arg1 oligosaccharides 1595:1610 N-linked oligosaccharides 1586:1610 N-linked oligosaccharides disappeared 1586:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 62 theme C-terminal 806:815 arg1 region 817:822 the C-terminal region 802:822 the C-terminal region of C4ST-1 802:832 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 63 theme degrees 1033:1039 arg1 C 1041:1041 37 degrees C 1030:1041 37 degrees C 1030:1041 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 64 theme N-glycosidase 572:584 arg1 F 586:586 peptide N-glycosidase F 564:586 peptide N-glycosidase F 564:586 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 65 from decrease 604:611 arg1 activity 616:623 activity 616:623 activity 616:623 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 66 from region 1432:1437 arg1 oligosaccharide 1398:1412 the N-linked oligosaccharide 1385:1412 the N-linked oligosaccharide at the C-terminal region 1385:1437 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 67 theme wildtype 1532:1539 arg1 protein 1541:1547 the wildtype protein 1528:1547 the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared 1528:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 68 from region 817:822 arg1 located 791:797 located 791:797 located 791:797 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 69 theme N-glycan 882:889 arg1 attachment 859:868 (iv) attachment 854:868 (iv) attachment of a single N-glycan at the C-terminal region 854:914 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 70 theme N-linked 1726:1733 arg1 oligosaccharide 1735:1749 the N-linked oligosaccharide 1722:1749 the N-linked oligosaccharide at the C-terminal region 1722:1774 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 4 71 theme N-linked 463:470 arg1 oligosaccharides 472:487 the N-linked oligosaccharides 459:487 the N-linked oligosaccharides attached to C4ST-1 459:506 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 0 72 theme active 68:73 arg1 form 75:78 the active form 64:78 the active form of chondroitin 4-sulphotransferase-1 64:115 N-linked oligosaccharides are required to produce and stabilize the active form of chondroitin 4-sulphotransferase-1. 15628971 7 73 link N-linked 1726:1733 arg1 oligosaccharide 1735:1749 the N-linked oligosaccharide 1722:1749 the N-linked oligosaccharide at the C-terminal region 1722:1774 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 74 theme recombinant 540:550 arg1 C4ST-1 552:557 recombinant C4ST-1 540:557 recombinant C4ST-1 540:557 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 75 theme protein 1512:1518 arg1 band 1520:1523 a broad protein band 1504:1523 a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared 1504:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 3 76 theme % 369:369 arg1 oligosaccharides 380:395 35% N-linked oligosaccharides 367:395 35% N-linked oligosaccharides. 367:396 35% N-linked oligosaccharides. 15628971 7 77 link N-linked 1586:1593 arg1 oligosaccharides 1595:1610 N-linked oligosaccharides 1586:1610 N-linked oligosaccharides disappeared 1586:1622 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 78 theme oligosaccharides 1691:1706 arg1 numbers 1671:1677 different numbers 1661:1677 different numbers of N-linked oligosaccharides 1661:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 4 79 link N-linked 463:470 arg1 oligosaccharides 472:487 the N-linked oligosaccharides 459:487 the N-linked oligosaccharides attached to C4ST-1 459:506 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 5 80 theme total 1189:1193 arg1 loss 1195:1198 total loss 1189:1198 total loss of activity 1189:1210 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 0 81 theme 4-sulphotransferase-1 95:115 arg1 form 75:78 the active form 64:78 the active form of chondroitin 4-sulphotransferase-1 64:115 N-linked oligosaccharides are required to produce and stabilize the active form of chondroitin 4-sulphotransferase-1. 15628971 1 82 from position 183:190 arg1 chondroitin 222:232 chondroitin 222:232 chondroitin 222:232 C4ST-1 (chondroitin 4-sulphotransferase-1) transfers sulphate to position 4 of N-acetylgalactosamine in chondroitin. 15628971 7 83 gly glycosylation 1461:1473 arg1 C4ST 1498:1501 recombinant C4ST 1486:1501 recombinant C4ST 1486:1501 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 84 theme C4ST 1498:1501 arg1 pattern 1475:1481 the glycosylation pattern 1457:1481 the glycosylation pattern of recombinant C4ST 1457:1501 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 85 theme different 1661:1669 arg1 numbers 1671:1677 different numbers 1661:1677 different numbers of N-linked oligosaccharides 1661:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 4 86 attach attached 489:496 arg2 oligosaccharides 472:487 the N-linked oligosaccharides 459:487 the N-linked oligosaccharides attached to C4ST-1 459:506 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 4 86 attach attached 489:496 arg1 C4ST-1 501:506 C4ST-1 501:506 C4ST-1 501:506 In the present paper, we investigated the functional role of the N-linked oligosaccharides attached to C4ST-1. 15628971 2 87 theme cells 323:327 arg1 medium 294:299 the culture medium 282:299 the culture medium of rat chondrosarcoma cells 282:327 We showed previously that purified C4ST-1 from the culture medium of rat chondrosarcoma cells was a glycoprotein containing approx. 15628971 5 88 theme control 1052:1058 arg1 protein 1072:1078 the control recombinant protein 1048:1078 the control recombinant protein 1048:1078 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 89 theme marked 597:602 arg1 decrease 604:611 a marked decrease 595:611 a marked decrease in activity 595:623 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 90 with band 1520:1523 arg1 numbers 1671:1677 different numbers 1661:1677 different numbers of N-linked oligosaccharides 1661:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 5 91 from C 1041:1041 arg1 enzyme 997:1002 the resulting recombinant enzyme 971:1002 the resulting recombinant enzyme 971:1002 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 91 from C 1041:1041 arg1 unstable 1018:1025 unstable 1018:1025 unstable 1018:1025 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 92 theme C-terminal 1103:1112 arg1 region 1114:1119 C-terminal region 1103:1119 C-terminal region 1103:1119 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 93 theme discrete 1633:1640 arg1 bands 1650:1654 four discrete protein bands 1628:1654 four discrete protein bands with different numbers of N-linked oligosaccharides 1628:1706 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 7 94 theme recombinant 1486:1496 arg1 C4ST 1498:1501 recombinant C4ST 1486:1501 recombinant C4ST 1486:1501 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 15628971 6 95 theme active 1349:1354 arg1 form 1356:1359 the active form 1345:1359 the active form of C4ST-1 1345:1369 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 5 96 theme resulting 975:983 arg1 enzyme 997:1002 the resulting recombinant enzyme 971:1002 the resulting recombinant enzyme 971:1002 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 96 theme resulting 975:983 arg1 unstable 1018:1025 unstable 1018:1025 unstable 1018:1025 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 97 dep production 631:640 arg1 ii 627:628 ii 627:628 ii 627:628 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 98 from region 909:914 arg1 attachment 859:868 (iv) attachment 854:868 (iv) attachment of a single N-glycan at the C-terminal region 854:914 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 99 with treatment 527:535 arg1 F 586:586 peptide N-glycosidase F 564:586 peptide N-glycosidase F 564:586 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 5 100 theme form 951:954 arg1 production 926:935 production 926:935 production of the active form of C4ST-1 926:964 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 6 101 link N-linked 1254:1261 arg1 oligosaccharides 1263:1278 N-linked oligosaccharides 1254:1278 N-linked oligosaccharides attached to C4ST-1 1254:1297 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 6 102 theme N-linked 1254:1261 arg1 oligosaccharides 1263:1278 N-linked oligosaccharides 1254:1278 N-linked oligosaccharides attached to C4ST-1 1254:1297 These observations strongly suggest that N-linked oligosaccharides attached to C4ST-1 contribute to the production and stability of the active form of C4ST-1. 15628971 5 103 theme C4ST-1 959:964 arg1 form 951:954 the active form 940:954 the active form of C4ST-1 940:964 We found that (i) treatment of recombinant C4ST-1 with peptide N-glycosidase F caused a marked decrease in activity, (ii) production of the active form of C4ST-1 by COS-7 cells transfected with cDNA of C4ST-1 was inhibited by tunicamycin, (iii) deletion of the N-glycosylation site located at the C-terminal region of C4ST-1 abolished activity, (iv) attachment of a single N-glycan at the C-terminal region supported production of the active form of C4ST-1, but the resulting recombinant enzyme was much more unstable at 37 degrees C than the control recombinant protein, and (v) truncation of C-terminal region up to the N-glycosylation site at the C-terminal region resulted in total loss of activity. 15628971 7 104 from region 1769:1774 arg1 oligosaccharide 1735:1749 the N-linked oligosaccharide 1722:1749 the N-linked oligosaccharide at the C-terminal region 1722:1774 In addition, the N-linked oligosaccharide at the C-terminal region appears to affect the glycosylation pattern of recombinant C4ST; a broad protein band of the wildtype protein resulting from microheterogeneity of N-linked oligosaccharides disappeared and four discrete protein bands with different numbers of N-linked oligosaccharides appeared when the N-linked oligosaccharide at the C-terminal region was deleted. 11533490 4 0 theme angstrom 674:681 arg1 closure 683:689 a 20 angstrom closure 669:689 a 20 angstrom closure between the membrane-proximal domains of the dimer 669:740 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 1 1 from regulation 182:191 arg1 important 154:162 important 154:162 important 154:162 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 5 2 theme intracellular 919:931 arg1 signaling 933:941 intracellular signaling 919:941 intracellular signaling by the natriuretic receptor family 919:976 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 1 3 theme blood 167:171 arg1 regulation 182:191 blood pressure regulation 167:191 blood pressure regulation through interaction with natriuretic cell-surface receptors 167:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 2 4 with receptor 417:424 arg1 CNP 455:457 CNP 455:457 CNP 455:457 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 4 with receptor 417:424 arg1 NP 476:477 a 22-amino acid NP 460:477 a 22-amino acid NP 460:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 5 5 theme molecular 856:864 arg1 spring 866:871 a molecular spring 854:871 a molecular spring 854:871 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 3 6 theme dimer 540:544 arg1 interface 518:526 the interface 514:526 the interface of an NPR-C dimer 514:544 A single CNP molecule is bound in the interface of an NPR-C dimer, resulting in asymmetric interactions between the hormone and the symmetrically related receptors. 11533490 4 7 theme dimer 736:740 arg1 domains 721:727 the membrane-proximal domains 699:727 the membrane-proximal domains of the dimer 699:740 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 4 7 theme dimer 736:740 arg1 dimer 736:740 the dimer 732:740 the dimer 732:740 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 5 8 theme linker 829:834 arg1 peptide 836:842 a linker peptide 827:842 a linker peptide acting as a molecular spring 827:871 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 3 9 theme single 482:487 arg1 molecule 493:500 A single CNP molecule 480:500 A single CNP molecule 480:500 A single CNP molecule is bound in the interface of an NPR-C dimer, resulting in asymmetric interactions between the hormone and the symmetrically related receptors. 11533490 1 10 from important 154:162 arg1 regulation 182:191 blood pressure regulation 167:191 blood pressure regulation through interaction with natriuretic cell-surface receptors 167:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 1 11 theme pressure 173:180 arg1 regulation 182:191 blood pressure regulation 167:191 blood pressure regulation through interaction with natriuretic cell-surface receptors 167:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 2 12 dep thermodynamics 284:297 arg1 the 264:266 the 264:266 the 264:266 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 5 13 theme allosteric 896:905 arg1 opening 764:770 the opening 760:770 the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, 760:872 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 5 13 theme allosteric 896:905 arg1 trigger 907:913 a conserved allosteric trigger 884:913 a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family 884:976 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 2 14 theme receptor 417:424 arg1 receptor 417:424 the unliganded human NP receptor 393:424 the unliganded human NP receptor (NPR-C) 393:432 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 14 theme receptor 417:424 arg1 complex 442:448 its complex 438:448 its complex with CNP, a 22-amino acid NP 438:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 14 theme receptor 417:424 arg1 domain 383:388 the extracellular domain 365:388 the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP 365:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 5 15 theme cleft 790:794 arg1 opening 764:770 the opening 760:770 the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, 760:872 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 5 15 theme cleft 790:794 arg1 trigger 907:913 a conserved allosteric trigger 884:913 a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family 884:976 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 1 16 theme Natriuretic 88:98 arg1 peptides 100:107 Natriuretic peptides 88:107 Natriuretic peptides (NPs) 88:113 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 1 16 theme Natriuretic 88:98 arg1 hormones 145:152 vasoactive cyclic-peptide hormones 119:152 vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors 119:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 1 16 theme Natriuretic 88:98 arg1 NPs 110:112 NPs 110:112 NPs 110:112 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 2 17 theme NP 414:415 arg1 receptor 417:424 the unliganded human NP receptor 393:424 the unliganded human NP receptor (NPR-C) 393:432 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 17 theme NP 414:415 arg1 NPR-C 427:431 NPR-C 427:431 NPR-C 427:431 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 0 18 theme Allosteric 0:9 arg1 activation 11:20 Allosteric activation 0:20 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 0:86 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 11533490 2 19 theme human 408:412 arg1 receptor 417:424 the unliganded human NP receptor 393:424 the unliganded human NP receptor (NPR-C) 393:432 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 19 theme human 408:412 arg1 NPR-C 427:431 NPR-C 427:431 NPR-C 427:431 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 5 20 theme conserved 886:894 arg1 opening 764:770 the opening 760:770 the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, 760:872 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 5 20 theme conserved 886:894 arg1 trigger 907:913 a conserved allosteric trigger 884:913 a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family 884:976 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 3 21 theme NPR-C 534:538 arg1 dimer 540:544 an NPR-C dimer 531:544 an NPR-C dimer 531:544 A single CNP molecule is bound in the interface of an NPR-C dimer, resulting in asymmetric interactions between the hormone and the symmetrically related receptors. 11533490 2 22 theme complex 442:448 arg1 receptor 417:424 the unliganded human NP receptor 393:424 the unliganded human NP receptor (NPR-C) 393:432 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 22 theme complex 442:448 arg1 complex 442:448 its complex 438:448 its complex with CNP, a 22-amino acid NP 438:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 22 theme complex 442:448 arg1 domain 383:388 the extracellular domain 365:388 the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP 365:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 23 theme domain 383:388 arg1 thermodynamics 284:297 hormone-binding thermodynamics 268:297 hormone-binding thermodynamics 268:297 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 23 theme domain 383:388 arg1 structures 311:320 crystal structures 303:320 crystal structures 303:320 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 24 theme 22-amino 462:469 arg1 CNP 455:457 CNP 455:457 CNP 455:457 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 24 theme 22-amino 462:469 arg1 NP 476:477 a 22-amino acid NP 460:477 a 22-amino acid NP 460:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 1 25 theme natriuretic 218:228 arg1 receptors 243:251 natriuretic cell-surface receptors 218:251 natriuretic cell-surface receptors 218:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 0 26 theme natriuretic 41:51 arg1 dimer 70:74 a spring-loaded natriuretic peptide receptor dimer 25:74 a spring-loaded natriuretic peptide receptor dimer 25:74 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 11533490 5 27 from trigger 907:913 arg1 monomer 751:757 each monomer 746:757 each monomer 746:757 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 1 28 theme cell-surface 230:241 arg1 receptors 243:251 natriuretic cell-surface receptors 218:251 natriuretic cell-surface receptors 218:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 2 29 theme acid 471:474 arg1 CNP 455:457 CNP 455:457 CNP 455:457 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 29 theme acid 471:474 arg1 NP 476:477 a 22-amino acid NP 460:477 a 22-amino acid NP 460:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 0 30 theme spring-loaded 27:39 arg1 dimer 70:74 a spring-loaded natriuretic peptide receptor dimer 25:74 a spring-loaded natriuretic peptide receptor dimer 25:74 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 11533490 0 31 theme receptor 61:68 arg1 dimer 70:74 a spring-loaded natriuretic peptide receptor dimer 25:74 a spring-loaded natriuretic peptide receptor dimer 25:74 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 11533490 5 32 theme interdomain 778:788 arg1 cleft 790:794 an interdomain cleft 775:794 an interdomain cleft 775:794 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 3 33 theme related 626:632 arg1 receptors 634:642 the symmetrically related receptors 608:642 the symmetrically related receptors 608:642 A single CNP molecule is bound in the interface of an NPR-C dimer, resulting in asymmetric interactions between the hormone and the symmetrically related receptors. 11533490 2 34 with complex 442:448 arg1 CNP 455:457 CNP 455:457 CNP 455:457 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 34 with complex 442:448 arg1 NP 476:477 a 22-amino acid NP 460:477 a 22-amino acid NP 460:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 35 theme extracellular 369:381 arg1 receptor 417:424 the unliganded human NP receptor 393:424 the unliganded human NP receptor (NPR-C) 393:432 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 35 theme extracellular 369:381 arg1 complex 442:448 its complex 438:448 its complex with CNP, a 22-amino acid NP 438:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 35 theme extracellular 369:381 arg1 domain 383:388 the extracellular domain 365:388 the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP 365:477 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 0 36 theme peptide 53:59 arg1 dimer 70:74 a spring-loaded natriuretic peptide receptor dimer 25:74 a spring-loaded natriuretic peptide receptor dimer 25:74 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 11533490 1 37 with interaction 201:211 arg1 receptors 243:251 natriuretic cell-surface receptors 218:251 natriuretic cell-surface receptors 218:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 3 38 theme asymmetric 560:569 arg1 interactions 571:582 asymmetric interactions 560:582 asymmetric interactions between the hormone and the symmetrically related receptors 560:642 A single CNP molecule is bound in the interface of an NPR-C dimer, resulting in asymmetric interactions between the hormone and the symmetrically related receptors. 11533490 3 39 theme CNP 489:491 arg1 molecule 493:500 A single CNP molecule 480:500 A single CNP molecule 480:500 A single CNP molecule is bound in the interface of an NPR-C dimer, resulting in asymmetric interactions between the hormone and the symmetrically related receptors. 11533490 4 40 theme membrane-proximal 703:719 arg1 domains 721:727 the membrane-proximal domains 699:727 the membrane-proximal domains of the dimer 699:740 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 4 40 theme membrane-proximal 703:719 arg1 dimer 736:740 the dimer 732:740 the dimer 732:740 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 5 41 theme receptor 962:969 arg1 family 971:976 the natriuretic receptor family 946:976 the natriuretic receptor family 946:976 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 2 42 theme crystal 303:309 arg1 structures 311:320 crystal structures 303:320 crystal structures 303:320 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 4 43 theme 20 671:672 arg1 angstrom 674:681 angstrom 674:681 angstrom 674:681 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 1 44 theme vasoactive 119:128 arg1 peptides 100:107 Natriuretic peptides 88:107 Natriuretic peptides (NPs) 88:113 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 1 44 theme vasoactive 119:128 arg1 hormones 145:152 vasoactive cyclic-peptide hormones 119:152 vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors 119:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 5 45 theme natriuretic 950:960 arg1 family 971:976 the natriuretic receptor family 946:976 the natriuretic receptor family 946:976 In each monomer, the opening of an interdomain cleft, which is tethered together by a linker peptide acting as a molecular spring, is likely a conserved allosteric trigger for intracellular signaling by the natriuretic receptor family. 11533490 1 46 theme cyclic-peptide 130:143 arg1 peptides 100:107 Natriuretic peptides 88:107 Natriuretic peptides (NPs) 88:113 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 1 46 theme cyclic-peptide 130:143 arg1 hormones 145:152 vasoactive cyclic-peptide hormones 119:152 vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors 119:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 2 47 theme unliganded 397:406 arg1 receptor 417:424 the unliganded human NP receptor 393:424 the unliganded human NP receptor (NPR-C) 393:432 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 47 theme unliganded 397:406 arg1 NPR-C 427:431 NPR-C 427:431 NPR-C 427:431 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 4 48 theme Hormone 645:651 arg1 binding 653:659 Hormone binding 645:659 Hormone binding 645:659 Hormone binding induces a 20 angstrom closure between the membrane-proximal domains of the dimer. 11533490 0 49 theme dimer 70:74 arg1 activation 11:20 Allosteric activation 0:20 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 0:86 Allosteric activation of a spring-loaded natriuretic peptide receptor dimer by hormone. 11533490 2 50 theme hormone-binding 268:282 arg1 thermodynamics 284:297 hormone-binding thermodynamics 268:297 hormone-binding thermodynamics 268:297 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 1 51 theme important 154:162 arg1 peptides 100:107 Natriuretic peptides 88:107 Natriuretic peptides (NPs) 88:113 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 1 51 theme important 154:162 arg1 hormones 145:152 vasoactive cyclic-peptide hormones 119:152 vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors 119:251 Natriuretic peptides (NPs) are vasoactive cyclic-peptide hormones important in blood pressure regulation through interaction with natriuretic cell-surface receptors. 11533490 2 52 from angstroms 337:345 arg1 thermodynamics 284:297 hormone-binding thermodynamics 268:297 hormone-binding thermodynamics 268:297 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 11533490 2 52 from angstroms 337:345 arg1 structures 311:320 crystal structures 303:320 crystal structures 303:320 We report the hormone-binding thermodynamics and crystal structures at 2.9 and 2.0 angstroms, respectively, of the extracellular domain of the unliganded human NP receptor (NPR-C) and its complex with CNP, a 22-amino acid NP. 8576151 11 0 dep gelatinases 1890:1900 arg1 gelatinases 1890:1900 the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3 1886:2051 the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3 1886:2051 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 0 dep gelatinases 1890:1900 arg1 B. 1908:1909 B. 1908:1909 B. 1908:1909 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 0 dep gelatinases 1890:1900 arg1 A 1902:1902 A 1902:1902 A 1902:1902 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 7 1 theme Further 948:954 arg1 processing 956:965 Further processing 948:965 Further processing 948:965 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 7 2 theme r 1050:1050 arg1 48,000 1053:1058 M(r) 48,000 1048:1058 M(r) 48,000 1048:1058 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 7 2 theme r 1050:1050 arg1 enzyme 1040:1045 the final active enzyme 1023:1045 the final active enzyme (M(r) 48,000) 1023:1059 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 4 3 from role 427:430 arg1 turnover 470:477 connective tissue turnover 452:477 connective tissue turnover 452:477 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 8576151 12 4 theme significant 2116:2126 arg1 role 2128:2131 a significant role 2114:2131 a significant role 2114:2131 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 3 5 theme tumor 339:343 arg1 progression 345:355 tumor progression 339:355 tumor progression 339:355 269, 16766-16773), and a potential role in tumor progression has been proposed for this enzyme. 8576151 11 6 dep B. 1908:1909 arg1 inhibited 1936:1944 inhibited 1936:1944 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3 1932:2051 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 12 7 theme constituents 2177:2188 arg1 turnover 2140:2147 the turnover 2136:2147 the turnover of connective tissue matrix constituents 2136:2188 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 7 8 theme final 1027:1031 arg1 48,000 1053:1058 M(r) 48,000 1048:1058 M(r) 48,000 1048:1058 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 7 8 theme final 1027:1031 arg1 enzyme 1040:1045 the final active enzyme 1023:1045 the final active enzyme (M(r) 48,000) 1023:1059 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 9 9 theme type 1327:1330 arg1 collagen 1335:1342 soluble type II collagen 1319:1342 soluble type II collagen 1319:1342 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 10 10 theme neutrophil 1547:1556 arg1 MMP-8 1582:1586 MMP-8 1582:1586 MMP-8 1582:1586 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 10 theme neutrophil 1547:1556 arg1 MMP-1 1572:1576 MMP-1 1572:1576 MMP-1 1572:1576 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 10 theme neutrophil 1547:1556 arg1 collagenases 1558:1569 the fibroblast and neutrophil collagenases 1528:1569 the fibroblast and neutrophil collagenases (MMP-1 and MMP-8) 1528:1587 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 8 11 theme C-terminal 1164:1173 arg1 domain 1175:1180 the C-terminal domain 1160:1180 the C-terminal domain 1160:1180 Trypsin activation of procollagenase-3 also generated a Tyr85 N terminus, but it was evident that the C-terminal domain was rapidly lost, and hence the collagenolytic activity diminished. 8576151 4 12 theme possible 418:425 arg1 role 427:430 the possible role 414:430 the possible role of collagenase-3 in connective tissue turnover 414:477 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 8576151 7 13 theme active 1033:1038 arg1 48,000 1053:1058 M(r) 48,000 1048:1058 M(r) 48,000 1048:1058 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 7 13 theme active 1033:1038 arg1 enzyme 1040:1045 the final active enzyme 1023:1045 the final active enzyme (M(r) 48,000) 1023:1059 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 12 14 theme in 2081:2082 arg1 collagenase-3 2089:2101 in vivo collagenase-3 2081:2101 in vivo collagenase-3 2081:2101 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 10 15 theme comparable 1503:1512 arg1 efficiency 1514:1523 comparable efficiency 1503:1523 comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8) 1503:1587 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 8 16 theme Trypsin 1062:1068 arg1 activation 1070:1079 Trypsin activation 1062:1079 Trypsin activation of procollagenase-3 1062:1099 Trypsin activation of procollagenase-3 also generated a Tyr85 N terminus, but it was evident that the C-terminal domain was rapidly lost, and hence the collagenolytic activity diminished. 8576151 4 17 theme human 523:527 arg1 procollagenase-3 529:544 recombinant human procollagenase-3 511:544 recombinant human procollagenase-3 511:544 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 8576151 1 18 attach isolated 131:138 arg2 cDNA 57:60 The cDNA 53:60 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) 53:120 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 18 attach isolated 131:138 arg1 library 160:166 a breast tumor library 145:166 a breast tumor library 145:166 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 10 19 dep collagenases 1558:1569 arg1 MMP-8 1582:1586 MMP-8 1582:1586 MMP-8 1582:1586 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 19 dep collagenases 1558:1569 arg1 MMP-1 1572:1576 MMP-1 1572:1576 MMP-1 1572:1576 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 19 dep collagenases 1558:1569 arg1 collagenases 1558:1569 the fibroblast and neutrophil collagenases 1528:1569 the fibroblast and neutrophil collagenases (MMP-1 and MMP-8) 1528:1587 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 20 theme Fibrillar 1460:1468 arg1 collagen 1477:1484 Fibrillar type I collagen 1460:1484 Fibrillar type I collagen 1460:1484 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 1 21 theme novel 67:71 arg1 collagenase-3 99:111 collagenase-3 99:111 collagenase-3 (MMP-13) 99:120 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 21 theme novel 67:71 arg1 metalloproteinase 80:96 a novel matrix metalloproteinase 65:96 a novel matrix metalloproteinase 65:96 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 22 theme tumor 154:158 arg1 library 160:166 a breast tumor library 145:166 a breast tumor library 145:166 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 5 23 theme purified 594:601 arg1 procollagenase-3 603:618 The purified procollagenase-3 590:618 The purified procollagenase-3 590:618 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 9 24 theme soluble 1319:1325 arg1 collagen 1335:1342 soluble type II collagen 1319:1342 soluble type II collagen 1319:1342 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 1 25 theme matrix 73:78 arg1 collagenase-3 99:111 collagenase-3 99:111 collagenase-3 (MMP-13) 99:120 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 25 theme matrix 73:78 arg1 metalloproteinase 80:96 a novel matrix metalloproteinase 65:96 a novel matrix metalloproteinase 65:96 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 5 26 theme N-terminal 685:694 arg1 LPLPSGGD 711:718 LPLPSGGD 711:718 LPLPSGGD 711:718 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 5 26 theme N-terminal 685:694 arg1 sequence 696:703 the N-terminal sequence 681:703 the N-terminal sequence 681:703 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 7 27 theme peptide 1007:1013 arg1 bond 1015:1018 the Glu84-Tyr85 peptide bond 991:1018 the Glu84-Tyr85 peptide bond 991:1018 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 5 28 with consistent 730:739 arg1 sequence 765:772 the cDNA-predicted sequence 746:772 the cDNA-predicted sequence 746:772 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 1 29 theme metalloproteinase 80:96 arg1 cDNA 57:60 The cDNA 53:60 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) 53:120 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 0 30 theme Biochemical 0:10 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization of human collagenase-3. 0:51 Biochemical characterization of human collagenase-3. 8576151 6 31 theme p-aminophenylmercuric 806:826 arg1 acetate 828:834 p-aminophenylmercuric acetate 806:834 p-aminophenylmercuric acetate 806:834 The proenzyme was activated by p-aminophenylmercuric acetate or stromelysin, yielding an intermediate form of M(r) 50,000, which displayed the N-terminal sequence L58EVTGK. 8576151 11 32 theme stoichiometric 1955:1968 arg1 fashion 1970:1976 a 1:1 stoichiometric fashion 1949:1976 a 1:1 stoichiometric fashion 1949:1976 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 10 33 theme I 1475:1475 arg1 collagen 1477:1484 Fibrillar type I collagen 1460:1484 Fibrillar type I collagen 1460:1484 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 11 34 dep substrates 1655:1664 arg1 substrates 1655:1664 the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1643:1740 the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1643:1740 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 34 dep substrates 1655:1664 arg1 Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1706:1740 Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1706:1740 Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1706:1740 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 34 dep substrates 1655:1664 arg1 Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 1666:1700 Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 1666:1700 Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 1666:1700 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 0 35 theme human 32:36 arg1 collagenase-3 38:50 human collagenase-3 32:50 human collagenase-3 32:50 Biochemical characterization of human collagenase-3. 8576151 10 36 theme type 1470:1473 arg1 collagen 1477:1484 Fibrillar type I collagen 1460:1484 Fibrillar type I collagen 1460:1484 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 3 37 from role 331:334 arg1 progression 345:355 tumor progression 339:355 tumor progression 339:355 269, 16766-16773), and a potential role in tumor progression has been proposed for this enzyme. 8576151 1 38 dep isolated 131:138 arg1 Dicz-Itza 187:195 Dicz-Itza 187:195 Dicz-Itza 187:195 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 J. 251:252 J. 251:252 J. 251:252 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 I. 198:199 I. 198:199 I. 198:199 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 López-Otin 259:268 López-Otin 259:268 López-Otin 259:268 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 J. 177:178 J. 177:178 J. 177:178 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 R. 238:239 R. 238:239 R. 238:239 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 L. 223:224 L. 223:224 L. 223:224 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 M. 180:181 M. 180:181 M. 180:181 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 Tolivia 242:248 Tolivia 242:248 Tolivia 242:248 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 Balbin 202:207 Balbin 202:207 Balbin 202:207 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 C 271:271 C 271:271 C 271:271 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 M. 226:227 M. 226:227 M. 226:227 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 38 dep isolated 131:138 arg1 M. 210:211 M. 210:211 M. 210:211 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 4 39 theme tissue 463:468 arg1 turnover 470:477 connective tissue turnover 452:477 connective tissue turnover 452:477 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 8576151 7 40 theme Glu84-Tyr85 995:1005 arg1 bond 1015:1018 the Glu84-Tyr85 peptide bond 991:1018 the Glu84-Tyr85 peptide bond 991:1018 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 12 41 dep in 2081:2082 arg1 vivo 2084:2087 vivo 2084:2087 vivo 2084:2087 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 10 42 theme fibroblast 1532:1541 arg1 MMP-8 1582:1586 MMP-8 1582:1586 MMP-8 1582:1586 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 42 theme fibroblast 1532:1541 arg1 MMP-1 1572:1576 MMP-1 1572:1576 MMP-1 1572:1576 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 10 42 theme fibroblast 1532:1541 arg1 collagenases 1558:1569 the fibroblast and neutrophil collagenases 1528:1569 the fibroblast and neutrophil collagenases (MMP-1 and MMP-8) 1528:1587 Fibrillar type I collagen was cleaved with comparable efficiency to the fibroblast and neutrophil collagenases (MMP-1 and MMP-8), respectively. 8576151 12 43 theme connective 2152:2161 arg1 constituents 2177:2188 connective tissue matrix constituents 2152:2188 connective tissue matrix constituents 2152:2188 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 1 44 theme breast 147:152 arg1 library 160:166 a breast tumor library 145:166 a breast tumor library 145:166 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 4 45 theme connective 452:461 arg1 turnover 470:477 connective tissue turnover 452:477 connective tissue turnover 452:477 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 8576151 9 46 theme collagenase-3 1291:1303 arg1 specificity 1276:1286 the substrate specificity 1262:1286 the substrate specificity of collagenase-3 1262:1303 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 6 47 theme M 885:885 arg1 r 887:887 M(r) 50,000 885:895 M(r) 50,000 885:895 The proenzyme was activated by p-aminophenylmercuric acetate or stromelysin, yielding an intermediate form of M(r) 50,000, which displayed the N-terminal sequence L58EVTGK. 8576151 5 48 theme 60,000 673:678 arg1 M 665:665 a M 663:665 a M(r) of 60,000 663:678 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 5 48 theme 60,000 673:678 arg1 r 667:667 r 667:667 r 667:667 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 0 49 theme collagenase-3 38:50 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization of human collagenase-3. 0:51 Biochemical characterization of human collagenase-3. 8576151 11 50 theme peptide 1647:1653 arg1 substrates 1655:1664 the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1643:1740 the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1643:1740 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 50 theme peptide 1647:1653 arg1 Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1706:1740 Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1706:1740 Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 1706:1740 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 50 theme peptide 1647:1653 arg1 Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 1666:1700 Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 1666:1700 Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 1666:1700 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 51 theme active 1835:1840 arg1 sequence 1847:1854 the active site sequence 1831:1854 the active site sequence of collagenase-3 (MMP-13) 1831:1880 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 8 52 theme Tyr85 1118:1122 arg1 terminus 1126:1133 a Tyr85 N terminus 1116:1133 a Tyr85 N terminus 1116:1133 Trypsin activation of procollagenase-3 also generated a Tyr85 N terminus, but it was evident that the C-terminal domain was rapidly lost, and hence the collagenolytic activity diminished. 8576151 9 53 theme specificity 1276:1286 arg1 Analysis 1250:1257 Analysis 1250:1257 Analysis of the substrate specificity of collagenase-3 1250:1303 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 11 54 theme metalloproteinases 2006:2023 arg1 inhibitors 1992:2001 the tissue inhibitors 1981:2001 the tissue inhibitors 1981:2001 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 54 theme metalloproteinases 2006:2023 arg1 TIMP-2 2034:2039 TIMP-2 2034:2039 TIMP-2 2034:2039 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 54 theme metalloproteinases 2006:2023 arg1 TIMP-3 2046:2051 TIMP-3 2046:2051 TIMP-3 2046:2051 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 54 theme metalloproteinases 2006:2023 arg1 TIMP-1 2026:2031 TIMP-1 2026:2031 TIMP-1 2026:2031 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 55 theme Active 1911:1916 arg1 collagenase-3 1918:1930 Active collagenase-3 1911:1930 Active collagenase-3 1911:1930 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 12 56 theme tissue 2163:2168 arg1 constituents 2177:2188 connective tissue matrix constituents 2152:2188 connective tissue matrix constituents 2152:2188 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 9 57 theme substrate 1266:1274 arg1 specificity 1276:1286 the substrate specificity 1262:1286 the substrate specificity of collagenase-3 1262:1303 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 6 58 theme sequence 929:936 arg1 L58EVTGK 938:945 the N-terminal sequence L58EVTGK 914:945 the N-terminal sequence L58EVTGK 914:945 The proenzyme was activated by p-aminophenylmercuric acetate or stromelysin, yielding an intermediate form of M(r) 50,000, which displayed the N-terminal sequence L58EVTGK. 8576151 3 59 theme potential 321:329 arg1 role 331:334 a potential role 319:334 a potential role in tumor progression 319:355 269, 16766-16773), and a potential role in tumor progression has been proposed for this enzyme. 8576151 12 60 theme matrix 2170:2175 arg1 constituents 2177:2188 connective tissue matrix constituents 2152:2188 connective tissue matrix constituents 2152:2188 These results suggest that in vivo collagenase-3 could play a significant role in the turnover of connective tissue matrix constituents. 8576151 8 61 theme procollagenase-3 1084:1099 arg1 activation 1070:1079 Trypsin activation 1062:1079 Trypsin activation of procollagenase-3 1062:1099 Trypsin activation of procollagenase-3 also generated a Tyr85 N terminus, but it was evident that the C-terminal domain was rapidly lost, and hence the collagenolytic activity diminished. 8576151 1 62 dep M. 180:181 arg1 Biol 284:287 Biol 284:287 Biol 284:287 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 1 62 dep M. 180:181 arg1 P. 183:184 Freije, J. M. P. 169:184 P. 183:184 The cDNA of a novel matrix metalloproteinase, collagenase-3 (MMP-13) has been isolated from a breast tumor library (Freije, J. M. P., Dicz-Itza, I., Balbin, M., Sanchez, L. M., Blasco, R., Tolivia, J., and López-Otin, C. (1994) J. Biol. 8576151 5 63 gly glycosylated 636:647 arg1 procollagenase-3 603:618 The purified procollagenase-3 590:618 The purified procollagenase-3 590:618 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 11 64 theme collagenase-3 1859:1871 arg1 sequence 1847:1854 the active site sequence 1831:1854 the active site sequence of collagenase-3 (MMP-13) 1831:1880 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 64 theme collagenase-3 1859:1871 arg1 gelatinases 1890:1900 the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3 1886:2051 the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3 1886:2051 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 64 theme collagenase-3 1859:1871 arg1 A 1902:1902 A 1902:1902 A 1902:1902 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 64 theme collagenase-3 1859:1871 arg1 B. 1908:1909 B. 1908:1909 B. 1908:1909 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 65 theme site 1842:1845 arg1 sequence 1847:1854 the active site sequence 1831:1854 the active site sequence of collagenase-3 (MMP-13) 1831:1880 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 8 66 theme N 1124:1124 arg1 terminus 1126:1133 a Tyr85 N terminus 1116:1133 a Tyr85 N terminus 1116:1133 Trypsin activation of procollagenase-3 also generated a Tyr85 N terminus, but it was evident that the C-terminal domain was rapidly lost, and hence the collagenolytic activity diminished. 8576151 5 67 theme cDNA-predicted 750:763 arg1 sequence 765:772 the cDNA-predicted sequence 746:772 the cDNA-predicted sequence 746:772 The purified procollagenase-3 was shown to be glycosylated and displayed a M(r) of 60,000, the N-terminal sequence being LPLPSGGD, which is consistent with the cDNA-predicted sequence. 8576151 6 68 theme intermediate 864:875 arg1 form 877:880 an intermediate form 861:880 an intermediate form 861:880 The proenzyme was activated by p-aminophenylmercuric acetate or stromelysin, yielding an intermediate form of M(r) 50,000, which displayed the N-terminal sequence L58EVTGK. 8576151 7 69 theme M 1048:1048 arg1 48,000 1053:1058 M(r) 48,000 1048:1058 M(r) 48,000 1048:1058 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 7 69 theme M 1048:1048 arg1 enzyme 1040:1045 the final active enzyme 1023:1045 the final active enzyme (M(r) 48,000) 1023:1059 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 9 70 dep efficient 1414:1422 arg1 efficient 1414:1422 efficient 1414:1422 efficient 1414:1422 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 9 70 dep efficient 1414:1422 arg1 times 1403:1407 5 or 6 times 1396:1407 5 or 6 times 1396:1407 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 9 70 dep efficient 1414:1422 arg1 enzyme 1385:1390 the enzyme 1381:1390 the enzyme 1381:1390 Analysis of the substrate specificity of collagenase-3 revealed that soluble type II collagen was preferentially hydrolyzed, while the enzyme was 5 or 6 times less efficient at cleaving type I or III collagen. 8576151 8 71 theme collagenolytic 1214:1227 arg1 activity 1229:1236 hence the collagenolytic activity 1204:1236 hence the collagenolytic activity 1204:1236 Trypsin activation of procollagenase-3 also generated a Tyr85 N terminus, but it was evident that the C-terminal domain was rapidly lost, and hence the collagenolytic activity diminished. 8576151 6 72 theme r 887:887 arg1 form 877:880 an intermediate form 861:880 an intermediate form 861:880 The proenzyme was activated by p-aminophenylmercuric acetate or stromelysin, yielding an intermediate form of M(r) 50,000, which displayed the N-terminal sequence L58EVTGK. 8576151 4 73 theme recombinant 511:521 arg1 procollagenase-3 529:544 recombinant human procollagenase-3 511:544 recombinant human procollagenase-3 511:544 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 8576151 11 74 theme tissue 1985:1990 arg1 inhibitors 1992:2001 the tissue inhibitors 1981:2001 the tissue inhibitors 1981:2001 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 74 theme tissue 1985:1990 arg1 TIMP-2 2034:2039 TIMP-2 2034:2039 TIMP-2 2034:2039 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 74 theme tissue 1985:1990 arg1 TIMP-3 2046:2051 TIMP-3 2046:2051 TIMP-3 2046:2051 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 11 74 theme tissue 1985:1990 arg1 TIMP-1 2026:2031 TIMP-1 2026:2031 TIMP-1 2026:2031 Unlike these collagenases, gelatin and the peptide substrates Mea-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 and Mca-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 were efficiently hydrolyzed as well, as would be predicted from the similarities between the active site sequence of collagenase-3 (MMP-13) and the gelatinases A and B. Active collagenase-3 was inhibited in a 1:1 stoichiometric fashion by the tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and TIMP-3. 8576151 6 75 theme N-terminal 918:927 arg1 L58EVTGK 938:945 the N-terminal sequence L58EVTGK 914:945 the N-terminal sequence L58EVTGK 914:945 The proenzyme was activated by p-aminophenylmercuric acetate or stromelysin, yielding an intermediate form of M(r) 50,000, which displayed the N-terminal sequence L58EVTGK. 8576151 7 76 theme bond 1015:1018 arg1 cleavage 979:986 cleavage 979:986 cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000) 979:1059 Further processing resulted in cleavage of the Glu84-Tyr85 peptide bond to the final active enzyme (M(r) 48,000). 8576151 4 77 theme collagenase-3 435:447 arg1 role 427:430 the possible role 414:430 the possible role of collagenase-3 in connective tissue turnover 414:477 In order to establish the possible role of collagenase-3 in connective tissue turnover, we have expressed and purified recombinant human procollagenase-3 and characterized the enzyme biochemically. 10358049 0 0 theme lipase 70:75 arg1 enzymes 92:98 two lipolytic enzymes 78:98 two lipolytic enzymes of medical interest 78:118 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 0 theme lipase 70:75 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human gastric lipase 0:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 0 theme lipase 70:75 arg1 model 46:50 model 46:50 model of lysosomal acid lipase 46:75 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 3 1 theme serine 627:632 arg1 carboxypeptidases 634:650 serine carboxypeptidases 627:650 serine carboxypeptidases 627:650 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 4 2 theme Leu-67 769:774 arg1 Leu-67 769:774 Leu-67 769:774 Leu-67 769:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 2 theme Leu-67 769:774 arg1 Gln-154 757:763 Gln-154 757:763 Gln-154 757:763 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 2 theme Leu-67 769:774 arg1 groups 747:752 NH groups 744:752 NH groups of Gln-154 and Leu-67 744:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 3 3 theme globular 462:469 arg1 enzyme 471:476 This globular enzyme 457:476 This globular enzyme (379 residues) 457:491 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 3 theme globular 462:469 arg1 residues 483:490 379 residues 479:490 379 residues 479:490 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 0 4 theme acid 65:68 arg1 lipase 70:75 lysosomal acid lipase 55:75 lysosomal acid lipase 55:75 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 9 5 theme reported 1315:1322 arg1 mutations 1324:1332 some previously reported mutations 1299:1332 some previously reported mutations leading to the cholesterol ester storage disease 1299:1381 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 9 6 theme cholesterol 1349:1359 arg1 disease 1375:1381 the cholesterol ester storage disease 1345:1381 the cholesterol ester storage disease 1345:1381 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 5 7 theme N-glycosylation 783:797 arg1 sites 799:803 Four N-glycosylation sites 778:803 Four N-glycosylation sites 778:803 Four N-glycosylation sites were identified on the electron density maps. 10358049 4 8 theme catalytic 678:686 arg1 triad 688:692 a classical catalytic triad 666:692 a classical catalytic triad (Ser-153, His-353, Asp-324) 666:720 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 9 9 theme possible 1273:1280 arg1 explanations 1282:1293 possible explanations 1273:1293 possible explanations for some previously reported mutations leading to the cholesterol ester storage disease 1273:1381 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 5 10 theme density 837:843 arg1 maps 845:848 the electron density maps 824:848 the electron density maps 824:848 Four N-glycosylation sites were identified on the electron density maps. 10358049 9 11 theme present 1406:1412 arg1 model 1414:1418 the present model 1402:1418 the present model 1402:1418 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 8 12 theme substrate 1194:1202 arg1 site 1212:1215 the hydrophobic substrate binding site 1178:1215 the hydrophobic substrate binding site 1178:1215 A phosphonate inhibitor was positioned in the active site that clearly suggests the location of the hydrophobic substrate binding site. 10358049 6 13 theme cap 983:985 arg1 domain 987:992 the cap domain 979:992 the cap domain 979:992 The catalytic serine is deeply buried under a segment consisting of 30 residues, which can be defined as a lid and belonging to the cap domain. 10358049 3 14 attach present 616:622 arg2 that 611:614 that 611:614 that 611:614 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 14 attach present 616:622 arg1 carboxypeptidases 634:650 serine carboxypeptidases 627:650 serine carboxypeptidases 627:650 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 4 15 contain possesses 656:664 arg2 triad 688:692 a classical catalytic triad 666:692 a classical catalytic triad (Ser-153, His-353, Asp-324) 666:720 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 15 contain possesses 656:664 arg2 hole 738:741 an oxyanion hole 726:741 an oxyanion hole (NH groups of Gln-154 and Leu-67) 726:775 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 15 contain possesses 656:664 arg1 It 653:654 It 653:654 It 653:654 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 1 16 from digestion 125:133 arg1 humans 138:143 humans 138:143 humans 138:143 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 2 17 theme 3.0-A 362:366 arg1 resolution 368:377 3.0-A resolution 362:377 3.0-A resolution 362:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 0 18 theme lipolytic 82:90 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human gastric lipase 0:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 18 theme lipolytic 82:90 arg1 enzymes 92:98 two lipolytic enzymes 78:98 two lipolytic enzymes of medical interest 78:118 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 2 19 theme acid 437:440 arg1 family 449:454 the mammalian acid lipase family 423:454 the mammalian acid lipase family 423:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 8 20 theme site 1212:1215 arg1 location 1166:1173 the location 1162:1173 the location of the hydrophobic substrate binding site 1162:1215 A phosphonate inhibitor was positioned in the active site that clearly suggests the location of the hydrophobic substrate binding site. 10358049 8 21 theme phosphonate 1084:1094 arg1 inhibitor 1096:1104 A phosphonate inhibitor 1082:1104 A phosphonate inhibitor 1082:1104 A phosphonate inhibitor was positioned in the active site that clearly suggests the location of the hydrophobic substrate binding site. 10358049 2 22 theme mammalian 427:435 arg1 family 449:454 the mammalian acid lipase family 423:454 the mammalian acid lipase family 423:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 23 theme lipase 442:447 arg1 family 449:454 the mammalian acid lipase family 423:454 the mammalian acid lipase family 423:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 4 24 theme Gln-154 757:763 arg1 Leu-67 769:774 Leu-67 769:774 Leu-67 769:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 24 theme Gln-154 757:763 arg1 Gln-154 757:763 Gln-154 757:763 Gln-154 757:763 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 24 theme Gln-154 757:763 arg1 groups 747:752 NH groups 744:752 NH groups of Gln-154 and Leu-67 744:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 2 25 from structure 313:321 arg1 resolution 368:377 3.0-A resolution 362:377 3.0-A resolution 362:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 4 26 theme classical 668:676 arg1 triad 688:692 a classical catalytic triad 666:692 a classical catalytic triad (Ser-153, His-353, Asp-324) 666:720 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 2 27 theme lipase 352:357 arg1 structure 313:321 the structure 309:321 the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family 309:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 0 28 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human gastric lipase 0:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 28 theme Crystal 0:6 arg1 enzymes 92:98 two lipolytic enzymes 78:98 two lipolytic enzymes of medical interest 78:118 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 7 29 theme lid 1019:1021 arg1 necessary 1026:1034 necessary 1026:1034 necessary 1026:1034 The displacement of the lid is necessary for the substrates to have access to Ser-153. 10358049 7 29 theme lid 1019:1021 arg1 displacement 999:1010 The displacement 995:1010 The displacement of the lid 995:1021 The displacement of the lid is necessary for the substrates to have access to Ser-153. 10358049 9 30 theme acid 1232:1235 arg1 lipase 1237:1242 The lysosomal acid lipase 1218:1242 The lysosomal acid lipase 1218:1242 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 2 31 theme gastric 344:350 arg1 structure 390:398 the first structure 380:398 the first structure to be described within the mammalian acid lipase family 380:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 31 theme gastric 344:350 arg1 lipase 352:357 recombinant human gastric lipase 326:357 recombinant human gastric lipase at 3.0-A resolution 326:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 0 32 theme human 21:25 arg1 lipase 35:40 human gastric lipase 21:40 human gastric lipase 21:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 33 theme medical 103:109 arg1 interest 111:118 medical interest 103:118 medical interest 103:118 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 4 34 theme oxyanion 729:736 arg1 hole 738:741 an oxyanion hole 726:741 an oxyanion hole (NH groups of Gln-154 and Leu-67) 726:775 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 1 35 theme classical 167:175 arg1 lipase 188:193 the classical pancreatic lipase 163:193 the classical pancreatic lipase 163:193 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 2 36 from resolution 368:377 arg1 structure 390:398 the first structure 380:398 the first structure to be described within the mammalian acid lipase family 380:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 36 from resolution 368:377 arg1 lipase 352:357 recombinant human gastric lipase 326:357 recombinant human gastric lipase at 3.0-A resolution 326:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 36 from resolution 368:377 arg1 structure 313:321 the structure 309:321 the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family 309:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 37 theme human 338:342 arg1 structure 390:398 the first structure 380:398 the first structure to be described within the mammalian acid lipase family 380:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 37 theme human 338:342 arg1 lipase 352:357 recombinant human gastric lipase 326:357 recombinant human gastric lipase at 3.0-A resolution 326:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 3 38 theme core 507:510 arg1 analogous 598:606 analogous 598:606 analogous 598:606 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 38 theme core 507:510 arg1 domain 512:517 a core domain 505:517 a core domain belonging to the alpha/beta hydrolase-fold family 505:567 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 1 39 theme pancreatic 177:186 arg1 lipase 188:193 the classical pancreatic lipase 163:193 the classical pancreatic lipase 163:193 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 0 40 theme lipase 35:40 arg1 enzymes 92:98 two lipolytic enzymes 78:98 two lipolytic enzymes of medical interest 78:118 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 40 theme lipase 35:40 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human gastric lipase 0:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 40 theme lipase 35:40 arg1 model 46:50 model 46:50 model of lysosomal acid lipase 46:75 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 6 41 theme catalytic 855:863 arg1 serine 865:870 The catalytic serine 851:870 The catalytic serine 851:870 The catalytic serine is deeply buried under a segment consisting of 30 residues, which can be defined as a lid and belonging to the cap domain. 10358049 3 42 from present 616:622 arg1 carboxypeptidases 634:650 serine carboxypeptidases 627:650 serine carboxypeptidases 627:650 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 43 theme alpha/beta 536:545 arg1 family 562:567 the alpha/beta hydrolase-fold family 532:567 the alpha/beta hydrolase-fold family 532:567 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 44 from carboxypeptidases 634:650 arg1 present 616:622 present 616:622 present 616:622 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 0 45 theme gastric 27:33 arg1 lipase 35:40 human gastric lipase 21:40 human gastric lipase 21:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 1 46 theme acidic 266:271 arg1 environment 281:291 the highly acidic stomach environment 255:291 the highly acidic stomach environment 255:291 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 0 47 theme interest 111:118 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human gastric lipase 0:40 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 0 47 theme interest 111:118 arg1 enzymes 92:98 two lipolytic enzymes 78:98 two lipolytic enzymes of medical interest 78:118 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 3 48 theme cap 576:578 arg1 domain 581:586 a "cap" domain 573:586 a "cap" domain 573:586 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 48 theme cap 576:578 arg1 analogous 598:606 analogous 598:606 analogous 598:606 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 49 theme hydrolase-fold 547:560 arg1 family 562:567 the alpha/beta hydrolase-fold family 532:567 the alpha/beta hydrolase-fold family 532:567 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 8 50 theme active 1128:1133 arg1 site 1135:1138 the active site 1124:1138 the active site that clearly suggests the location of the hydrophobic substrate binding site 1124:1215 A phosphonate inhibitor was positioned in the active site that clearly suggests the location of the hydrophobic substrate binding site. 10358049 1 51 theme stomach 273:279 arg1 environment 281:291 the highly acidic stomach environment 255:291 the highly acidic stomach environment 255:291 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 5 52 gly N-glycosylation 783:797 arg2 Four 778:781 Four 778:781 Four 778:781 Four N-glycosylation sites were identified on the electron density maps. 10358049 5 52 gly N-glycosylation 783:797 arg2 sites 799:803 Four N-glycosylation sites 778:803 Four N-glycosylation sites 778:803 Four N-glycosylation sites were identified on the electron density maps. 10358049 3 53 theme " 579:579 arg1 domain 581:586 a "cap" domain 573:586 a "cap" domain 573:586 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 3 53 theme " 579:579 arg1 analogous 598:606 analogous 598:606 analogous 598:606 This globular enzyme (379 residues) consists of a core domain belonging to the alpha/beta hydrolase-fold family and a "cap" domain, which is analogous to that present in serine carboxypeptidases. 10358049 4 54 dep triad 688:692 arg1 Ser-153 695:701 Ser-153 695:701 Ser-153 695:701 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 54 dep triad 688:692 arg1 Asp-324 713:719 Asp-324 713:719 Asp-324 713:719 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 54 dep triad 688:692 arg1 His-353 704:710 His-353 704:710 His-353 704:710 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 55 theme NH 744:745 arg1 Leu-67 769:774 Leu-67 769:774 Leu-67 769:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 55 theme NH 744:745 arg1 Gln-154 757:763 Gln-154 757:763 Gln-154 757:763 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 55 theme NH 744:745 arg1 groups 747:752 NH groups 744:752 NH groups of Gln-154 and Leu-67 744:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 56 dep hole 738:741 arg1 Leu-67 769:774 Leu-67 769:774 Leu-67 769:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 56 dep hole 738:741 arg1 Gln-154 757:763 Gln-154 757:763 Gln-154 757:763 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 4 56 dep hole 738:741 arg1 groups 747:752 NH groups 744:752 NH groups of Gln-154 and Leu-67 744:774 It possesses a classical catalytic triad (Ser-153, His-353, Asp-324) and an oxyanion hole (NH groups of Gln-154 and Leu-67). 10358049 2 57 dep structure 390:398 arg1 described 406:414 described 406:414 to be described within the mammalian acid lipase family 400:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 9 58 theme ester 1361:1365 arg1 disease 1375:1381 the cholesterol ester storage disease 1345:1381 the cholesterol ester storage disease 1345:1381 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 7 59 contain have 1058:1061 arg1 necessary 1026:1034 necessary 1026:1034 necessary 1026:1034 The displacement of the lid is necessary for the substrates to have access to Ser-153. 10358049 7 59 contain have 1058:1061 arg1 displacement 999:1010 The displacement 995:1010 The displacement of the lid 995:1021 The displacement of the lid is necessary for the substrates to have access to Ser-153. 10358049 7 59 contain have 1058:1061 arg2 access 1063:1068 access 1063:1068 access to Ser-153 1063:1079 The displacement of the lid is necessary for the substrates to have access to Ser-153. 10358049 9 60 theme storage 1367:1373 arg1 disease 1375:1381 the cholesterol ester storage disease 1345:1381 the cholesterol ester storage disease 1345:1381 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10358049 8 61 theme binding 1204:1210 arg1 site 1212:1215 the hydrophobic substrate binding site 1178:1215 the hydrophobic substrate binding site 1178:1215 A phosphonate inhibitor was positioned in the active site that clearly suggests the location of the hydrophobic substrate binding site. 10358049 5 62 theme electron 828:835 arg1 maps 845:848 the electron density maps 824:848 the electron density maps 824:848 Four N-glycosylation sites were identified on the electron density maps. 10358049 8 63 theme hydrophobic 1182:1192 arg1 site 1212:1215 the hydrophobic substrate binding site 1178:1215 the hydrophobic substrate binding site 1178:1215 A phosphonate inhibitor was positioned in the active site that clearly suggests the location of the hydrophobic substrate binding site. 10358049 1 64 theme gastric 204:210 arg1 lipase 212:217 gastric lipase 204:217 gastric lipase 204:217 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 0 65 theme lysosomal 55:63 arg1 lipase 70:75 lysosomal acid lipase 55:75 lysosomal acid lipase 55:75 Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest. 10358049 2 66 theme first 384:388 arg1 structure 390:398 the first structure 380:398 the first structure to be described within the mammalian acid lipase family 380:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 66 theme first 384:388 arg1 lipase 352:357 recombinant human gastric lipase 326:357 recombinant human gastric lipase at 3.0-A resolution 326:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 1 67 theme Fat 121:123 arg1 digestion 125:133 Fat digestion 121:133 Fat digestion in humans 121:143 Fat digestion in humans requires not only the classical pancreatic lipase but also gastric lipase, which is stable and active despite the highly acidic stomach environment. 10358049 2 68 theme recombinant 326:336 arg1 structure 390:398 the first structure 380:398 the first structure to be described within the mammalian acid lipase family 380:454 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 2 68 theme recombinant 326:336 arg1 lipase 352:357 recombinant human gastric lipase 326:357 recombinant human gastric lipase at 3.0-A resolution 326:377 We report here the structure of recombinant human gastric lipase at 3.0-A resolution, the first structure to be described within the mammalian acid lipase family. 10358049 9 69 theme lysosomal 1222:1230 arg1 lipase 1237:1242 The lysosomal acid lipase 1218:1242 The lysosomal acid lipase 1218:1242 The lysosomal acid lipase was modeled by homology, and possible explanations for some previously reported mutations leading to the cholesterol ester storage disease are given based on the present model. 10821832 4 0 theme luminal 1041:1047 arg1 loops 1049:1053 3 large and 4 small luminal loops 1021:1053 3 large and 4 small luminal loops 1021:1053 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 5 1 theme putative 1143:1150 arg1 those 1198:1202 those 1198:1202 those 1198:1202 Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG-R and SCAP, providing strong evidence that this domain is functionally important. 10821832 5 1 theme putative 1143:1150 arg1 SSD 1152:1154 the putative SSD 1139:1154 the putative SSD of NPC1 1139:1162 Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG-R and SCAP, providing strong evidence that this domain is functionally important. 10821832 0 2 theme regulatory 201:210 arg1 protein 256:262 sterol regulatory element binding protein cleavage-activating protein 194:262 sterol regulatory element binding protein cleavage-activating protein 194:262 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 3 with region 363:368 arg1 homology 385:392 extensive homology 375:392 extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP) 375:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 4 theme NPC1 286:289 arg1 glycoprotein 331:342 a polytopic glycoprotein 319:342 a polytopic glycoprotein 319:342 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 4 theme NPC1 286:289 arg1 protein 292:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 0 5 theme sterol 194:199 arg1 protein 256:262 sterol regulatory element binding protein cleavage-activating protein 194:262 sterol regulatory element binding protein cleavage-activating protein 194:262 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 6 theme A 469:469 arg1 HMG-R 482:486 HMG-R 482:486 HMG-R 482:486 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 6 theme A 469:469 arg1 reductase 471:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase 433:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) 433:487 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 4 7 theme cytoplasmic 1089:1099 arg1 tail 1101:1104 a cytoplasmic tail 1087:1104 a cytoplasmic tail 1087:1104 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 0 8 theme putative 93:100 arg1 domain 117:122 the putative sterol-sensing domain 89:122 the putative sterol-sensing domain 89:122 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 0 9 theme binding 220:226 arg1 protein 256:262 sterol regulatory element binding protein cleavage-activating protein 194:262 sterol regulatory element binding protein cleavage-activating protein 194:262 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 10 theme reductase 471:479 arg1 domains 416:422 the sterol-sensing domains 397:422 the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP) 397:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 10 theme reductase 471:479 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 10 theme reductase 471:479 arg1 reductase 471:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase 433:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) 433:487 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 10 theme reductase 471:479 arg1 SSD 425:427 SSD 425:427 SSD 425:427 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 3 11 theme glycosylated 864:875 arg1 domains 877:883 glycosylated domains 864:883 glycosylated domains that are located in the lumen of the endoplasmic reticulum 864:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 1 12 theme protein 555:561 arg1 domains 416:422 the sterol-sensing domains 397:422 the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP) 397:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 12 theme protein 555:561 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 12 theme protein 555:561 arg1 reductase 471:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase 433:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) 433:487 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 12 theme protein 555:561 arg1 SSD 425:427 SSD 425:427 SSD 425:427 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 0 13 theme element 212:218 arg1 protein 256:262 sterol regulatory element binding protein cleavage-activating protein 194:262 sterol regulatory element binding protein cleavage-activating protein 194:262 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 3 14 theme domains 877:883 arg1 identification 846:859 identification 846:859 identification 846:859 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 3 14 theme domains 877:883 arg1 NPC1 837:840 NPC1 837:840 NPC1 837:840 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 0 15 theme domain 117:122 arg1 orientation 74:84 the membrane orientation 61:84 the membrane orientation of the putative sterol-sensing domain 61:122 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 0 15 theme domain 117:122 arg1 identical 127:135 identical 127:135 identical 127:135 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 16 theme extensive 375:383 arg1 homology 385:392 extensive homology 375:392 extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP) 375:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 0 17 theme cleavage-activating 236:254 arg1 protein 256:262 sterol regulatory element binding protein cleavage-activating protein 194:262 sterol regulatory element binding protein cleavage-activating protein 194:262 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 3 18 theme reticulum 934:942 arg1 lumen 909:913 the lumen 905:913 the lumen of the endoplasmic reticulum 905:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 0 19 theme sterol-sensing 102:115 arg1 domain 117:122 the putative sterol-sensing domain 89:122 the putative sterol-sensing domain 89:122 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 0 20 theme protein 228:234 arg1 protein 256:262 sterol regulatory element binding protein cleavage-activating protein 194:262 sterol regulatory element binding protein cleavage-activating protein 194:262 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 2 21 theme investigation 708:720 arg1 expression 661:670 expression 661:670 expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells 661:780 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 4 22 theme small 1058:1062 arg1 loops 1076:1080 6 small cytoplasmic loops 1056:1080 6 small cytoplasmic loops 1056:1080 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 2 23 theme topology 635:642 arg1 model 621:625 a model 619:625 a model of NPC1 topology 619:642 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 4 24 theme large 1023:1027 arg1 loops 1049:1053 3 large and 4 small luminal loops 1021:1053 3 large and 4 small luminal loops 1021:1053 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 3 25 from located 894:900 arg1 lumen 909:913 the lumen 905:913 the lumen of the endoplasmic reticulum 905:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 0 26 theme Topological 0:10 arg1 analysis 12:19 Topological analysis 0:19 Topological analysis of Niemann-Pick C1 protein 0:46 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 2 27 theme NPC1 630:633 arg1 topology 635:642 NPC1 topology 630:642 NPC1 topology 630:642 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 1 28 theme sterol 493:498 arg1 SCAP 564:567 SCAP 564:567 SCAP 564:567 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 28 theme sterol 493:498 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 2 29 theme accessibility 733:745 arg1 proteins 695:702 epitope-tagged NPC1 proteins 675:702 epitope-tagged NPC1 proteins 675:702 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 2 29 theme accessibility 733:745 arg1 investigation 708:720 investigation 708:720 investigation of epitope accessibility 708:745 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 0 30 theme Niemann-Pick 24:35 arg1 protein 40:46 Niemann-Pick C1 protein 24:46 Niemann-Pick C1 protein 24:46 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 2 31 theme proteins 695:702 arg1 expression 661:670 expression 661:670 expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells 661:780 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 1 32 theme regulatory 500:509 arg1 SCAP 564:567 SCAP 564:567 SCAP 564:567 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 32 theme regulatory 500:509 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 2 33 theme epitope-tagged 675:688 arg1 proteins 695:702 epitope-tagged NPC1 proteins 675:702 epitope-tagged NPC1 proteins 675:702 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 4 34 theme cytoplasmic 1064:1074 arg1 loops 1076:1080 6 small cytoplasmic loops 1056:1080 6 small cytoplasmic loops 1056:1080 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 2 35 theme epitope 725:731 arg1 accessibility 733:745 epitope accessibility 725:745 epitope accessibility 725:745 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 3 36 from lumen 909:913 arg1 located 894:900 located 894:900 located 894:900 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 0 37 theme protein 40:46 arg1 analysis 12:19 Topological analysis 0:19 Topological analysis of Niemann-Pick C1 protein 0:46 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 38 theme sterol-sensing 401:414 arg1 domains 416:422 the sterol-sensing domains 397:422 the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP) 397:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 38 theme sterol-sensing 401:414 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 38 theme sterol-sensing 401:414 arg1 reductase 471:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase 433:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) 433:487 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 38 theme sterol-sensing 401:414 arg1 SSD 425:427 SSD 425:427 SSD 425:427 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 0 39 theme C1 37:38 arg1 protein 40:46 Niemann-Pick C1 protein 24:46 Niemann-Pick C1 protein 24:46 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 40 theme polytopic 321:329 arg1 glycoprotein 331:342 a polytopic glycoprotein 319:342 a polytopic glycoprotein 319:342 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 40 theme polytopic 321:329 arg1 protein 292:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 4 41 theme small 1035:1039 arg1 loops 1049:1053 3 large and 4 small luminal loops 1021:1053 3 large and 4 small luminal loops 1021:1053 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 3 42 theme NPC1 837:840 arg1 expression 823:832 expression 823:832 expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum 823:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 1 43 theme element 511:517 arg1 SCAP 564:567 SCAP 564:567 SCAP 564:567 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 43 theme element 511:517 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 4 44 theme transmembrane 998:1010 arg1 domains 1012:1018 13 transmembrane domains 995:1018 13 transmembrane domains 995:1018 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 0 45 theme 3-hydroxy-3-methylglutaryl-CoA 149:178 arg1 reductase 180:188 3-hydroxy-3-methylglutaryl-CoA reductase 149:188 3-hydroxy-3-methylglutaryl-CoA reductase 149:188 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 3 46 gly glycosylated 864:875 arg1 domains 877:883 glycosylated domains 864:883 glycosylated domains that are located in the lumen of the endoplasmic reticulum 864:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 1 47 theme binding 519:525 arg1 SCAP 564:567 SCAP 564:567 SCAP 564:567 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 47 theme binding 519:525 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 2 48 from expression 661:670 arg1 cells 776:780 selectively permeabilized cells 750:780 selectively permeabilized cells 750:780 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 5 49 theme same 1183:1186 arg1 manner 1188:1193 the same manner 1179:1193 the same manner 1179:1193 Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG-R and SCAP, providing strong evidence that this domain is functionally important. 10821832 4 50 gly glycoprotein 973:984 arg1 glycoprotein 973:984 this glycoprotein 968:984 this glycoprotein 968:984 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 1 51 theme protein 527:533 arg1 SCAP 564:567 SCAP 564:567 SCAP 564:567 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 51 theme protein 527:533 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 5 52 theme NPC1 1159:1162 arg1 those 1198:1202 those 1198:1202 those 1198:1202 Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG-R and SCAP, providing strong evidence that this domain is functionally important. 10821832 5 52 theme NPC1 1159:1162 arg1 SSD 1152:1154 the putative SSD 1139:1154 the putative SSD of NPC1 1139:1162 Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG-R and SCAP, providing strong evidence that this domain is functionally important. 10821832 2 53 theme NPC1 690:693 arg1 proteins 695:702 epitope-tagged NPC1 proteins 675:702 epitope-tagged NPC1 proteins 675:702 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 1 54 theme cleavage-activating 535:553 arg1 SCAP 564:567 SCAP 564:567 SCAP 564:567 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 54 theme cleavage-activating 535:553 arg1 protein 555:561 sterol regulatory element binding protein cleavage-activating protein 493:561 sterol regulatory element binding protein cleavage-activating protein (SCAP) 493:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 55 contain contains 352:359 arg1 it 349:350 it 349:350 it 349:350 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 55 contain contains 352:359 arg2 region 363:368 a region 361:368 a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP) 361:568 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 3 56 theme identification 846:859 arg1 expression 823:832 expression 823:832 expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum 823:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 2 57 theme functional 582:591 arg1 characterization 593:608 the functional characterization 578:608 the functional characterization of NPC1 578:616 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 2 58 theme NPC1 613:616 arg1 characterization 593:608 the functional characterization 578:608 the functional characterization of NPC1 578:616 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 1 59 gly glycoprotein 331:342 arg1 glycoprotein 331:342 a polytopic glycoprotein 319:342 a polytopic glycoprotein 319:342 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 59 gly glycoprotein 331:342 arg1 protein 292:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 4 60 contain contains 986:993 arg2 loops 1076:1080 6 small cytoplasmic loops 1056:1080 6 small cytoplasmic loops 1056:1080 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 4 60 contain contains 986:993 arg2 domains 1012:1018 13 transmembrane domains 995:1018 13 transmembrane domains 995:1018 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 4 60 contain contains 986:993 arg1 glycoprotein 973:984 this glycoprotein 968:984 this glycoprotein 968:984 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 4 60 contain contains 986:993 arg2 tail 1101:1104 a cytoplasmic tail 1087:1104 a cytoplasmic tail 1087:1104 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 4 60 contain contains 986:993 arg2 loops 1049:1053 3 large and 4 small luminal loops 1021:1053 3 large and 4 small luminal loops 1021:1053 Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. 10821832 1 61 theme Niemann-Pick 269:280 arg1 glycoprotein 331:342 a polytopic glycoprotein 319:342 a polytopic glycoprotein 319:342 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 61 theme Niemann-Pick 269:280 arg1 protein 292:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 0 62 theme membrane 65:72 arg1 orientation 74:84 the membrane orientation 61:84 the membrane orientation of the putative sterol-sensing domain 61:122 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 0 62 theme membrane 65:72 arg1 identical 127:135 identical 127:135 identical 127:135 Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. 10821832 1 63 theme 3-hydroxy-3-methylglutaryl-coenzyme 433:467 arg1 HMG-R 482:486 HMG-R 482:486 HMG-R 482:486 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 63 theme 3-hydroxy-3-methylglutaryl-coenzyme 433:467 arg1 reductase 471:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase 433:479 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) 433:487 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 64 theme C1 282:283 arg1 glycoprotein 331:342 a polytopic glycoprotein 319:342 a polytopic glycoprotein 319:342 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 1 64 theme C1 282:283 arg1 protein 292:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein 265:298 The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). 10821832 3 65 located located 894:900 arg2 domains 877:883 glycosylated domains 864:883 glycosylated domains that are located in the lumen of the endoplasmic reticulum 864:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 3 65 located located 894:900 arg1 lumen 909:913 the lumen 905:913 the lumen of the endoplasmic reticulum 905:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 2 66 theme permeabilized 762:774 arg1 cells 776:780 selectively permeabilized cells 750:780 selectively permeabilized cells 750:780 To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. 10821832 3 67 theme endoplasmic 922:932 arg1 reticulum 934:942 the endoplasmic reticulum 918:942 the endoplasmic reticulum 918:942 These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. 10821832 5 68 theme strong 1233:1238 arg1 evidence 1240:1247 strong evidence that this domain is functionally important 1233:1290 strong evidence that this domain is functionally important 1233:1290 Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG-R and SCAP, providing strong evidence that this domain is functionally important. 1400872 2 0 theme polymerase 487:496 arg1 reaction 504:511 a polymerase chain reaction to obtain the eight exons encoding SHBG 485:551 a polymerase chain reaction to obtain the eight exons encoding SHBG 485:551 This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG. 1400872 1 1 theme 48K 292:294 arg1 forms 269:273 three molecular weight forms 246:273 three molecular weight forms of 56K, 52K, and 48K 246:294 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 7 2 theme CHO 1646:1648 arg1 cells 1650:1654 CHO cells 1646:1654 CHO cells 1646:1654 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 7 3 theme sulfate-polyacrylamide 1423:1444 arg1 electrophoresis 1450:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 5 4 theme Chinese 1084:1090 arg1 cells 1112:1116 Chinese hamster ovary (CHO) cells 1084:1116 Chinese hamster ovary (CHO) cells 1084:1116 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 4 5 from position 889:896 arg1 site 856:859 an additional consensus site 832:859 an additional consensus site for N-glycosylation at this position 832:896 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 8 6 theme additional 1674:1683 arg1 subunit 1685:1691 This additional subunit 1669:1691 This additional subunit 1669:1691 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 5 7 theme expression 1044:1053 arg1 vector 1055:1060 the pRc/CMV expression vector 1032:1060 the pRc/CMV expression vector 1032:1060 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 7 8 theme gel 1446:1448 arg1 electrophoresis 1450:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 0 9 theme additional 84:93 arg1 chain 108:112 an additional carbohydrate chain 81:112 an additional carbohydrate chain 81:112 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 5 10 theme ovary 1100:1104 arg1 cells 1112:1116 Chinese hamster ovary (CHO) cells 1084:1116 Chinese hamster ovary (CHO) cells 1084:1116 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 4 11 theme consensus 846:854 arg1 site 856:859 an additional consensus site 832:859 an additional consensus site for N-glycosylation at this position 832:896 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 6 12 theme those 1325:1329 arg1 serum 1286:1290 the serum 1282:1290 the serum of either a normal individual or those who produce an electrophoretic variant (98%) 1282:1374 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 7 13 theme sodium 1408:1413 arg1 electrophoresis 1450:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 3 14 from substitution 636:647 arg1 polypeptide 690:700 the SHBG polypeptide 681:700 the SHBG polypeptide 681:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 4 15 theme additional 835:844 arg1 site 856:859 an additional consensus site 832:859 an additional consensus site for N-glycosylation at this position 832:896 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 9 16 theme CHO 1950:1952 arg1 cells 1954:1958 CHO cells 1950:1958 CHO cells 1950:1958 Nevertheless, its steroid-binding affinity was equal to normal SHBG produced by CHO cells or SHBG in serum. 1400872 4 17 theme SHBG 960:963 arg1 DNA 979:981 a human SHBG complementary DNA 952:981 a human SHBG complementary DNA 952:981 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 8 18 theme recombinant 1822:1832 arg1 SHBG 1834:1837 recombinant SHBG 1822:1837 recombinant SHBG 1822:1837 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 1 19 attach isolated 131:138 arg1 individual 148:157 an individual 145:157 an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 145:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 19 attach isolated 131:138 arg1 variant 219:225 homozygous for a sex hormone-binding globulin (SHBG) variant 166:225 homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 166:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 19 attach isolated 131:138 arg2 DNA 123:125 Genomic DNA 115:125 Genomic DNA 115:125 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 7 20 theme normal 1622:1627 arg1 SHBG 1629:1632 normal SHBG 1622:1632 normal SHBG produced by CHO cells or in serum 1622:1666 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 0 21 theme carbohydrate 95:106 arg1 chain 108:112 an additional carbohydrate chain 81:112 an additional carbohydrate chain 81:112 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 3 22 theme SHBG 685:688 arg1 polypeptide 690:700 the SHBG polypeptide 681:700 the SHBG polypeptide 681:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 23 from residue 666:672 arg1 Asn 658:660 Asp --> Asn 650:660 Asp --> Asn 650:660 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 23 from residue 666:672 arg1 substitution 636:647 an amino acid substitution 622:647 an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide 622:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 6 24 theme individual 1311:1320 arg1 serum 1286:1290 the serum 1282:1290 the serum of either a normal individual or those who produce an electrophoretic variant (98%) 1282:1374 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 8 25 from synthesis 1846:1854 arg1 cells 1863:1867 CHO cells 1859:1867 CHO cells 1859:1867 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 6 26 from SHBG 1274:1277 arg1 serum 1286:1290 the serum 1282:1290 the serum of either a normal individual or those who produce an electrophoretic variant (98%) 1282:1374 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 5 27 theme pRc/CMV 1036:1042 arg1 vector 1055:1060 the pRc/CMV expression vector 1032:1060 the pRc/CMV expression vector 1032:1060 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 7 28 theme CHO 1517:1519 arg1 cells 1521:1525 CHO cells 1517:1525 CHO cells 1517:1525 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 1 29 theme denaturing 325:334 arg1 conditions 336:345 denaturing conditions 325:345 denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 325:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 29 theme denaturing 325:334 arg1 electrophoresis 390:404 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 348:404 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 348:404 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 30 theme homozygous 166:175 arg1 variant 219:225 homozygous for a sex hormone-binding globulin (SHBG) variant 166:225 homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 166:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 30 theme homozygous 166:175 arg1 individual 148:157 an individual 145:157 an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 145:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 0 31 theme Molecular 0:8 arg1 analyses 10:17 Molecular analyses 0:17 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 0:113 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 1 32 theme molecular 252:260 arg1 forms 269:273 three molecular weight forms 246:273 three molecular weight forms of 56K, 52K, and 48K 246:294 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 9 33 theme normal 1926:1931 arg1 SHBG 1933:1936 normal SHBG 1926:1936 normal SHBG produced by CHO cells or SHBG in serum 1926:1975 Nevertheless, its steroid-binding affinity was equal to normal SHBG produced by CHO cells or SHBG in serum. 1400872 4 34 theme complementary 965:977 arg1 DNA 979:981 a human SHBG complementary DNA 952:981 a human SHBG complementary DNA 952:981 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 5 35 theme complementary 996:1008 arg1 DNA 1010:1012 The mutated complementary DNA 984:1012 The mutated complementary DNA 984:1012 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 8 36 theme complexity 1771:1780 arg1 degree 1761:1766 a greater degree 1751:1766 a greater degree of complexity 1751:1780 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 6 37 theme normal 1304:1309 arg1 individual 1311:1320 a normal individual 1302:1320 a normal individual 1302:1320 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 1 38 theme weight 262:267 arg1 forms 269:273 three molecular weight forms 246:273 three molecular weight forms of 56K, 52K, and 48K 246:294 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 7 39 theme dodecyl 1415:1421 arg1 electrophoresis 1450:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1408:1464 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 3 40 theme Sequence 554:561 arg1 analysis 563:570 Sequence analysis 554:570 Sequence analysis of these exons 554:585 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 7 41 theme 60K 1542:1544 arg1 subunit 1546:1552 a 60K subunit 1540:1552 a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum 1540:1666 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 6 42 from cells 1258:1262 arg1 serum 1286:1290 the serum 1282:1290 the serum of either a normal individual or those who produce an electrophoretic variant (98%) 1282:1374 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 6 43 theme CHO 1254:1256 arg1 % 1267:1267 85% 1265:1267 85% 1265:1267 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 6 43 theme CHO 1254:1256 arg1 cells 1258:1262 CHO cells 1254:1262 CHO cells (85%) 1254:1268 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 2 44 theme oligonucleotide 458:472 arg1 primers 474:480 intron-specific oligonucleotide primers 442:480 intron-specific oligonucleotide primers 442:480 This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG. 1400872 1 45 theme sex 183:185 arg1 SHBG 213:216 SHBG 213:216 SHBG 213:216 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 45 theme sex 183:185 arg1 globulin 203:210 a sex hormone-binding globulin 181:210 a sex hormone-binding globulin (SHBG) 181:217 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 0 46 theme sex 30:32 arg1 globulin 50:57 a human sex hormone-binding globulin 22:57 a human sex hormone-binding globulin variant 22:65 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 3 47 theme same 711:714 arg1 mutation 716:723 the same mutation 707:723 the same mutation 707:723 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 9 48 theme steroid-binding 1888:1902 arg1 affinity 1904:1911 its steroid-binding affinity 1884:1911 its steroid-binding affinity 1884:1911 Nevertheless, its steroid-binding affinity was equal to normal SHBG produced by CHO cells or SHBG in serum. 1400872 3 49 theme Asp 650:652 arg1 Asn 658:660 Asp --> Asn 650:660 Asp --> Asn 650:660 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 49 theme Asp 650:652 arg1 substitution 636:647 an amino acid substitution 622:647 an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide 622:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 1 50 theme hormone-binding 187:201 arg1 SHBG 213:216 SHBG 213:216 SHBG 213:216 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 50 theme hormone-binding 187:201 arg1 globulin 203:210 a sex hormone-binding globulin 181:210 a sex hormone-binding globulin (SHBG) 181:217 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 0 51 theme human 24:28 arg1 globulin 50:57 a human sex hormone-binding globulin 22:57 a human sex hormone-binding globulin variant 22:65 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 1 52 theme 56K 278:280 arg1 forms 269:273 three molecular weight forms 246:273 three molecular weight forms of 56K, 52K, and 48K 246:294 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 8 53 theme CHO 1859:1861 arg1 cells 1863:1867 CHO cells 1859:1867 CHO cells 1859:1867 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 1 54 theme sulfate-polyacrylamide 363:384 arg1 conditions 336:345 denaturing conditions 325:345 denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 325:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 54 theme sulfate-polyacrylamide 363:384 arg1 electrophoresis 390:404 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 348:404 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 348:404 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 3 55 theme point 598:602 arg1 mutation 604:611 a point mutation 596:611 a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide 596:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 8 56 theme carbohydrate 1789:1800 arg1 structures 1802:1811 the carbohydrate structures 1785:1811 the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells 1785:1867 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 0 57 theme globulin 50:57 arg1 variant 59:65 a human sex hormone-binding globulin variant 22:65 a human sex hormone-binding globulin variant 22:65 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 5 58 theme CHO 1107:1109 arg1 cells 1112:1116 Chinese hamster ovary (CHO) cells 1084:1116 Chinese hamster ovary (CHO) cells 1084:1116 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 8 59 theme greater 1753:1759 arg1 degree 1761:1766 a greater degree 1751:1766 a greater degree of complexity 1751:1780 This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells. 1400872 1 60 theme gel 386:388 arg1 conditions 336:345 denaturing conditions 325:345 denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 325:405 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 1 60 theme gel 386:388 arg1 electrophoresis 390:404 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 348:404 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 348:404 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 3 61 theme exons 581:585 arg1 analysis 563:570 Sequence analysis 554:570 Sequence analysis of these exons 554:585 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 5 62 theme hamster 1092:1098 arg1 cells 1112:1116 Chinese hamster ovary (CHO) cells 1084:1116 Chinese hamster ovary (CHO) cells 1084:1116 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 0 63 theme hormone-binding 34:48 arg1 globulin 50:57 a human sex hormone-binding globulin 22:57 a human sex hormone-binding globulin variant 22:65 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 1 64 theme 52K 283:285 arg1 forms 269:273 three molecular weight forms 246:273 three molecular weight forms of 56K, 52K, and 48K 246:294 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 6 65 theme normal 1230:1235 arg1 SHBG 1237:1240 normal SHBG 1230:1240 normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%) 1230:1374 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 2 66 theme intron-specific 442:456 arg1 primers 474:480 intron-specific oligonucleotide primers 442:480 intron-specific oligonucleotide primers 442:480 This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG. 1400872 5 67 theme mutated 988:994 arg1 DNA 1010:1012 The mutated complementary DNA 984:1012 The mutated complementary DNA 984:1012 The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells. 1400872 1 68 theme Genomic 115:121 arg1 DNA 123:125 Genomic DNA 115:125 Genomic DNA 115:125 Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). 1400872 7 69 theme light 1585:1589 arg1 subunits 1597:1604 the heavy (52K) and light (48K) subunits 1565:1604 a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum 1540:1666 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 4 70 dep introduced 933:942 arg1 confirm 906:912 confirm 906:912 to confirm its utilization 903:928 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 3 71 theme acid 631:634 arg1 Asn 658:660 Asp --> Asn 650:660 Asp --> Asn 650:660 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 71 theme acid 631:634 arg1 substitution 636:647 an amino acid substitution 622:647 an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide 622:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 7 72 theme Western 1470:1476 arg1 blotting 1478:1485 Western blotting 1470:1485 Western blotting 1470:1485 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 4 73 theme human 954:958 arg1 DNA 979:981 a human SHBG complementary DNA 952:981 a human SHBG complementary DNA 952:981 This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA. 1400872 3 74 theme --> 654:656 arg1 Asn 658:660 Asp --> Asn 650:660 Asp --> Asn 650:660 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 74 theme --> 654:656 arg1 substitution 636:647 an amino acid substitution 622:647 an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide 622:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 75 theme amino 625:629 arg1 Asn 658:660 Asp --> Asn 650:660 Asp --> Asn 650:660 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 3 75 theme amino 625:629 arg1 substitution 636:647 an amino acid substitution 622:647 an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide 622:700 Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait. 1400872 2 76 theme chain 498:502 arg1 reaction 504:511 a polymerase chain reaction to obtain the eight exons encoding SHBG 485:551 a polymerase chain reaction to obtain the eight exons encoding SHBG 485:551 This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG. 1400872 7 77 theme heavy 1569:1573 arg1 subunits 1597:1604 the heavy (52K) and light (48K) subunits 1565:1604 a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum 1540:1666 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 0 78 dep analyses 10:17 arg1 evidence 68:75 evidence 68:75 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 0:113 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 7 79 theme SHBG 1492:1495 arg1 variant 1497:1503 the SHBG variant 1488:1503 the SHBG variant produced by CHO cells 1488:1525 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 6 80 theme electrophoretic 1346:1360 arg1 variant 1362:1368 an electrophoretic variant 1343:1368 an electrophoretic variant (98%) 1343:1374 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 6 80 theme electrophoretic 1346:1360 arg1 % 1373:1373 98% 1371:1373 98% 1371:1373 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 0 81 theme variant 59:65 arg1 analyses 10:17 Molecular analyses 0:17 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 0:113 Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain. 1400872 7 82 dep heavy 1569:1573 arg1 48K 1592:1594 48K 1592:1594 48K 1592:1594 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 7 82 dep heavy 1569:1573 arg1 52K 1576:1578 52K 1576:1578 52K 1576:1578 Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum. 1400872 6 83 dep less 1172:1175 arg1 % 1186:1186 54% 1184:1186 54% 1184:1186 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 1400872 6 84 theme concanavalin 1198:1209 arg1 A 1211:1211 concanavalin A 1198:1211 concanavalin A 1198:1211 The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%). 7918467 0 0 theme intramolecular 73:86 arg1 links 98:102 the intermolecular and intramolecular disulfide links 50:102 the intermolecular and intramolecular disulfide links 50:102 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 7 1 from analysis 615:622 arg1 cluster 638:644 a purified cluster 627:644 a purified cluster of chymotryptic fragments 627:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 9 2 theme intramolecular 914:927 arg1 linkages 939:946 two intramolecular disulfide linkages 910:946 two intramolecular disulfide linkages 910:946 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 1 3 theme human 149:153 arg1 apoD 173:176 apoD 173:176 apoD 173:176 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 1 3 theme human 149:153 arg1 D 170:170 human apolipoprotein D 149:170 human apolipoprotein D (apoD) 149:177 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 10 4 theme lipoprotein 1160:1170 arg1 surfaces 1172:1179 lipoprotein surfaces 1160:1179 lipoprotein surfaces 1160:1179 We conclude that apoD binds to lipoprotein surfaces through structures other than alpha-helices, such as disulfide links. 7918467 7 5 from combination 551:561 arg1 cluster 638:644 a purified cluster 627:644 a purified cluster of chymotryptic fragments 627:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 6 theme manual 566:571 arg1 degradations 579:590 manual Edman degradations 566:590 manual Edman degradations 566:590 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 7 theme mass 596:599 arg1 analysis 615:622 mass spectrometric analysis 596:622 mass spectrometric analysis on a purified cluster of chymotryptic fragments 596:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 1 8 theme apolipoprotein 155:168 arg1 apoD 173:176 apoD 173:176 apoD 173:176 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 1 8 theme apolipoprotein 155:168 arg1 D 170:170 human apolipoprotein D 149:170 human apolipoprotein D (apoD) 149:177 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 6 9 theme peptide 461:467 arg1 analysis 469:476 Tryptic peptide analysis 453:476 Tryptic peptide analysis 453:476 Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. 7918467 0 10 theme links 98:102 arg1 locations 37:45 locations 37:45 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 0:103 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 7 11 theme Edman 573:577 arg1 degradations 579:590 manual Edman degradations 566:590 manual Edman degradations 566:590 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 9 12 theme alpha-helical 993:1005 arg1 structures 1007:1016 alpha-helical structures 993:1016 alpha-helical structures 993:1016 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 7 13 theme chymotryptic 649:660 arg1 fragments 662:670 chymotryptic fragments 649:670 chymotryptic fragments 649:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 0 14 theme disulfide 88:96 arg1 links 98:102 the intermolecular and intramolecular disulfide links 50:102 the intermolecular and intramolecular disulfide links 50:102 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 2 15 dep [Drayna 332:338 arg1 al 343:344 et al 340:344 cDNA [Drayna et al. (1986) J. Biol 327:360 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 2 15 dep [Drayna 332:338 arg1 Biol 357:360 Biol 357:360 Biol 357:360 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 7 16 theme Cys6 802:805 arg1 Cys8 730:733 Cys8 730:733 Cys8 730:733 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 16 theme Cys6 802:805 arg1 bridge 768:773 an intermolecular bridge 750:773 an intermolecular bridge between Cys116 750:788 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 16 theme Cys6 802:805 arg1 Cys114 739:744 Cys114 739:744 Cys114 739:744 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 17 theme fragments 662:670 arg1 cluster 638:644 a purified cluster 627:644 a purified cluster of chymotryptic fragments 627:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 6 18 theme Tryptic 453:459 arg1 analysis 469:476 Tryptic peptide analysis 453:476 Tryptic peptide analysis 453:476 Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. 7918467 1 19 theme D 170:170 arg1 structure 136:144 the primary structure 124:144 the primary structure of human apolipoprotein D (apoD) 124:177 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 7 20 theme disulfide 705:713 arg1 bridge 715:720 an intramolecular disulfide bridge 687:720 an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II 687:816 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 21 theme apoD 793:796 arg1 Cys8 730:733 Cys8 730:733 Cys8 730:733 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 21 theme apoD 793:796 arg1 bridge 768:773 an intermolecular bridge 750:773 an intermolecular bridge between Cys116 750:788 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 21 theme apoD 793:796 arg1 Cys114 739:744 Cys114 739:744 Cys114 739:744 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 8 22 located found 862:866 arg1 Asn45 871:875 Asn45 871:875 Asn45 871:875 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 8 22 located found 862:866 arg1 Asn78 881:885 Asn78 881:885 Asn78 881:885 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 8 22 located found 862:866 arg2 sites 832:836 sites 832:836 sites of N-glycosylation 832:855 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 8 22 located found 862:866 arg1 addition 822:829 addition 822:829 addition 822:829 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 8 22 located found 862:866 arg2 N-glycosylation 841:855 N-glycosylation 841:855 N-glycosylation 841:855 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 2 23 attach derived 314:320 arg2 structure 304:312 the primary structure 292:312 the primary structure derived from cDNA [Drayna et al. (1986) J. Biol 292:360 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 2 23 attach derived 314:320 arg1 1986 348:351 1986 348:351 1986 348:351 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 2 23 attach derived 314:320 arg1 [Drayna 332:338 cDNA [Drayna 327:338 cDNA [Drayna et al. (1986) J. Biol 327:360 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 2 24 theme primary 296:302 arg1 structure 304:312 the primary structure 292:312 the primary structure derived from cDNA [Drayna et al. (1986) J. Biol 292:360 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 6 25 theme disulfide 525:533 arg1 bridge 535:540 a disulfide bridge 523:540 a disulfide bridge 523:540 Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. 7918467 7 26 theme intermolecular 753:766 arg1 bridge 768:773 an intermolecular bridge 750:773 an intermolecular bridge between Cys116 750:788 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 5 27 theme amino 409:413 arg1 residues 420:427 169 amino acid residues 405:427 169 amino acid residues 405:427 ApoD consists of 169 amino acid residues, including 5 cysteines. 7918467 5 27 theme amino 409:413 arg1 cysteines 442:450 5 cysteines 440:450 5 cysteines 440:450 ApoD consists of 169 amino acid residues, including 5 cysteines. 7918467 0 28 theme human 13:17 arg1 D 34:34 human apolipoprotein D 13:34 human apolipoprotein D 13:34 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 9 29 theme regions 1056:1062 arg1 formation 1019:1027 formation 1019:1027 formation of the amphipathic helical regions that characterize the other soluble apolipoproteins 1019:1114 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 5 30 theme acid 415:418 arg1 residues 420:427 169 amino acid residues 405:427 169 amino acid residues 405:427 ApoD consists of 169 amino acid residues, including 5 cysteines. 7918467 5 30 theme acid 415:418 arg1 cysteines 442:450 5 cysteines 440:450 5 cysteines 440:450 ApoD consists of 169 amino acid residues, including 5 cysteines. 7918467 9 31 theme disulfide 929:937 arg1 linkages 939:946 two intramolecular disulfide linkages 910:946 two intramolecular disulfide linkages 910:946 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 7 32 theme apoA-II 810:816 arg1 Cys6 802:805 Cys6 802:805 Cys6 802:805 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 32 theme apoA-II 810:816 arg1 apoD 793:796 apoD 793:796 apoD 793:796 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 8 33 theme N-glycosylation 841:855 arg1 sites 832:836 sites 832:836 sites of N-glycosylation 832:855 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 8 33 theme N-glycosylation 841:855 arg1 N-glycosylation 841:855 N-glycosylation 841:855 N-glycosylation 841:855 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 0 34 theme D 34:34 arg1 Structure 0:8 Structure 0:8 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 0:103 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 0 35 theme apolipoprotein 19:32 arg1 D 34:34 human apolipoprotein D 13:34 human apolipoprotein D 13:34 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 10 36 theme disulfide 1234:1242 arg1 links 1244:1248 disulfide links 1234:1248 disulfide links 1234:1248 We conclude that apoD binds to lipoprotein surfaces through structures other than alpha-helices, such as disulfide links. 7918467 7 37 theme purified 629:636 arg1 cluster 638:644 a purified cluster 627:644 a purified cluster of chymotryptic fragments 627:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 9 38 contain contains 901:908 arg1 apoD 896:899 apoD 896:899 apoD 896:899 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 9 38 contain contains 901:908 arg2 linkages 939:946 two intramolecular disulfide linkages 910:946 two intramolecular disulfide linkages 910:946 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 2 39 theme cDNA 327:330 arg1 1986 348:351 1986 348:351 1986 348:351 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 2 39 theme cDNA 327:330 arg1 [Drayna 332:338 cDNA [Drayna 327:338 cDNA [Drayna et al. (1986) J. Biol 327:360 Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. 7918467 9 40 theme amphipathic 1036:1046 arg1 regions 1056:1062 the amphipathic helical regions 1032:1062 the amphipathic helical regions that characterize the other soluble apolipoproteins 1032:1114 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 10 41 theme other 1200:1204 arg1 structures 1189:1198 structures 1189:1198 structures 1189:1198 We conclude that apoD binds to lipoprotein surfaces through structures other than alpha-helices, such as disulfide links. 7918467 10 41 theme other 1200:1204 arg1 links 1244:1248 disulfide links 1234:1248 disulfide links 1234:1248 We conclude that apoD binds to lipoprotein surfaces through structures other than alpha-helices, such as disulfide links. 7918467 9 42 contain has 952:954 arg1 apoD 896:899 apoD 896:899 apoD 896:899 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 9 42 contain has 952:954 arg2 content 963:969 a high content 956:969 a high content of proline to disrupt alpha-helical structures 956:1016 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 9 43 theme other 1086:1090 arg1 apolipoproteins 1100:1114 the other soluble apolipoproteins 1082:1114 the other soluble apolipoproteins 1082:1114 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 9 44 theme high 958:961 arg1 content 963:969 a high content 956:969 a high content of proline to disrupt alpha-helical structures 956:1016 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 7 45 theme intramolecular 690:703 arg1 bridge 715:720 an intramolecular disulfide bridge 687:720 an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II 687:816 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 8 46 gly N-glycosylation 841:855 arg2 N-glycosylation 841:855 N-glycosylation 841:855 N-glycosylation 841:855 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 8 46 gly N-glycosylation 841:855 arg2 sites 832:836 sites 832:836 sites of N-glycosylation 832:855 In addition, sites of N-glycosylation were found at Asn45 and Asn78. 7918467 1 47 attach derived 200:206 arg2 peptides 191:198 peptides 191:198 peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin 191:269 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 1 47 attach derived 200:206 arg1 digestions 213:222 digestions 213:222 digestions 213:222 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 9 48 theme soluble 1092:1098 arg1 apolipoproteins 1100:1114 the other soluble apolipoproteins 1082:1114 the other soluble apolipoproteins 1082:1114 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 1 49 theme primary 128:134 arg1 structure 136:144 the primary structure 124:144 the primary structure of human apolipoprotein D (apoD) 124:177 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 7918467 4 50 dep 16535-16539 374:384 arg1 261 369:371 261 369:371 261 369:371 261, 16535-16539]. 7918467 0 51 theme intermolecular 54:67 arg1 links 98:102 the intermolecular and intramolecular disulfide links 50:102 the intermolecular and intramolecular disulfide links 50:102 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 7 52 theme spectrometric 601:613 arg1 analysis 615:622 mass spectrometric analysis 596:622 mass spectrometric analysis on a purified cluster of chymotryptic fragments 596:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 9 53 theme helical 1048:1054 arg1 regions 1056:1062 the amphipathic helical regions 1032:1062 the amphipathic helical regions that characterize the other soluble apolipoproteins 1032:1114 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 7 54 theme degradations 579:590 arg1 combination 551:561 a combination 549:561 a combination of manual Edman degradations 549:590 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 7 54 theme degradations 579:590 arg1 analysis 615:622 mass spectrometric analysis 596:622 mass spectrometric analysis on a purified cluster of chymotryptic fragments 596:670 Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. 7918467 0 55 dep Structure 0:8 arg1 locations 37:45 locations 37:45 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 0:103 Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links. 7918467 9 56 theme proline 974:980 arg1 content 963:969 a high content 956:969 a high content of proline to disrupt alpha-helical structures 956:1016 Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. 7918467 1 57 theme cyanogen 227:234 arg1 bromide 236:242 cyanogen bromide 227:242 cyanogen bromide 227:242 We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. 2721499 7 0 theme relationship 1272:1283 arg1 evidence 1239:1246 the only evidence 1230:1246 the only evidence of a possible vestigial relationship between SP-40,40 and other complement components 1230:1332 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 2 1 theme mRNA 327:330 arg1 molecule 332:339 the same mRNA molecule 318:339 the same mRNA molecule 318:339 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 3 2 contain contains 557:564 arg2 sites 593:597 six N-linked glycosylation sites 566:597 six N-linked glycosylation sites distributed equally between the two chains of the structure 566:657 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 3 2 contain contains 557:564 arg1 precursor 489:497 The precursor 485:497 The precursor 485:497 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 0 3 theme protein 81:87 arg1 characterization 22:37 characterization 22:37 characterization 22:37 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 0 3 theme protein 81:87 arg1 cloning 10:16 cloning 10:16 cloning 10:16 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 2 4 from frame 309:313 arg1 molecule 332:339 the same mRNA molecule 318:339 the same mRNA molecule 318:339 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 2 5 theme same 322:325 arg1 molecule 332:339 the same mRNA molecule 318:339 the same mRNA molecule 318:339 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 4 6 theme % 709:709 arg1 identity 711:718 a 77% identity 705:718 a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 705:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 8 7 theme precise 1339:1345 arg1 role 1347:1350 The precise role 1335:1350 The precise role of SP-40,40 1335:1362 The precise role of SP-40,40 is not known in either blood or semen but the present findings document an intriguing link between the immune and the reproductive systems. 2721499 4 8 theme rat 725:727 arg1 product 790:796 the major secreted product 771:796 the major secreted product of Sertoli cells 771:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 8 theme rat 725:727 arg1 SGP-2 755:759 SGP-2 755:759 SGP-2 755:759 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 8 theme rat 725:727 arg1 glycoprotein-2 739:752 a rat sulphated glycoprotein-2 723:752 a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 723:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 6 9 theme SP-40,40 1064:1071 arg1 beta-chain 1050:1059 the beta-chain 1046:1059 the beta-chain of SP-40,40 1046:1071 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 3 10 theme vectorial 536:544 arg1 export 546:551 vectorial export 536:551 vectorial export 536:551 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 4 11 contain bears 699:703 arg1 sequence 664:671 The sequence 660:671 The sequence of the SP-40,40 precursor 660:697 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 11 contain bears 699:703 arg2 identity 711:718 a 77% identity 705:718 a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 705:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 6 12 dep components 1156:1165 arg1 C7 1167:1168 C7 1167:1168 C7 1167:1168 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 3 13 theme signal 516:521 arg1 sequence 523:530 a signal sequence 514:530 a signal sequence for vectorial export 514:551 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 5 14 theme comparable 879:888 arg1 levels 872:877 levels 872:877 levels comparable to those in serum 872:906 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 2 15 theme precursor 385:393 arg1 protein 395:401 a biosynthetic precursor protein 370:401 a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond 370:482 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 2 16 theme SP-40,40 265:272 arg1 chains 255:260 The two chains 247:260 The two chains of SP-40,40 247:272 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 1 17 theme cDNA 160:163 arg1 sequence 165:172 The cDNA sequence 156:172 The cDNA sequence encoding the human complement-associated protein, SP-40,40, 156:232 The cDNA sequence encoding the human complement-associated protein, SP-40,40, is reported. 2721499 3 18 link N-linked 570:577 arg1 sites 593:597 six N-linked glycosylation sites 566:597 six N-linked glycosylation sites distributed equally between the two chains of the structure 566:657 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 7 19 theme possible 1253:1260 arg1 relationship 1272:1283 a possible vestigial relationship 1251:1283 a possible vestigial relationship between SP-40,40 and other complement components 1251:1332 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 5 20 from levels 872:877 arg1 plasma 862:867 human seminal plasma 848:867 human seminal plasma at levels comparable to those in serum 848:906 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 4 21 theme sulphated 729:737 arg1 product 790:796 the major secreted product 771:796 the major secreted product of Sertoli cells 771:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 21 theme sulphated 729:737 arg1 SGP-2 755:759 SGP-2 755:759 SGP-2 755:759 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 21 theme sulphated 729:737 arg1 glycoprotein-2 739:752 a rat sulphated glycoprotein-2 723:752 a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 723:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 22 theme cells 809:813 arg1 product 790:796 the major secreted product 771:796 the major secreted product of Sertoli cells 771:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 22 theme cells 809:813 arg1 glycoprotein-2 739:752 a rat sulphated glycoprotein-2 723:752 a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 723:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 0 23 theme complement-associated 59:79 arg1 protein 81:87 the novel, human complement-associated protein 42:87 protein 81:87 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 0 23 theme complement-associated 59:79 arg1 SP-40,40 90:97 SP-40,40 90:97 SP-40,40 90:97 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 2 24 theme open 296:299 arg1 frame 309:313 a single open reading frame 287:313 a single open reading frame on the same mRNA molecule 287:339 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 7 25 theme vestigial 1262:1270 arg1 relationship 1272:1283 a possible vestigial relationship 1251:1283 a possible vestigial relationship between SP-40,40 and other complement components 1251:1332 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 0 26 theme Molecular 0:8 arg1 cloning 10:16 cloning 10:16 cloning 10:16 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 6 27 theme complement 1145:1154 arg1 components 1156:1165 complement components C7, C8 and C9 1145:1179 components 1156:1165 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 6 27 theme complement 1145:1154 arg1 C8 1171:1172 C8 1171:1172 C8 1171:1172 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 6 27 theme complement 1145:1154 arg1 C9 1178:1179 C9 1178:1179 C9 1178:1179 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 4 28 theme Sertoli 801:807 arg1 cells 809:813 Sertoli cells 801:813 Sertoli cells 801:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 2 29 theme single 289:294 arg1 frame 309:313 a single open reading frame 287:313 a single open reading frame on the same mRNA molecule 287:339 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 3 30 gly glycosylation 579:591 arg2 sites 593:597 six N-linked glycosylation sites 566:597 six N-linked glycosylation sites distributed equally between the two chains of the structure 566:657 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 3 30 gly glycosylation 579:591 arg2 six 566:568 six 566:568 six 566:568 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 3 31 theme glycosylation 579:591 arg1 sites 593:597 six N-linked glycosylation sites 566:597 six N-linked glycosylation sites distributed equally between the two chains of the structure 566:657 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 5 32 theme seminal 975:981 arg1 forms 983:987 serum and seminal forms 965:987 forms 983:987 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 32 theme seminal 975:981 arg1 SGP-2 955:959 SGP-2 955:959 SGP-2 955:959 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 32 theme seminal 975:981 arg1 SP-40,40 942:949 SP-40,40 942:949 SP-40,40 942:949 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 2 33 theme protein 395:401 arg1 existence 357:365 the existence 353:365 the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond 353:482 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 4 34 theme 77 707:708 arg1 identity 711:718 a 77% identity 705:718 a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 705:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 6 35 theme corresponding 1107:1119 arg1 segments 1121:1128 corresponding segments 1107:1128 corresponding segments located within complement components C7, C8 and C9 1107:1179 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 5 36 theme protein 1001:1007 arg1 forms 983:987 serum and seminal forms 965:987 forms 983:987 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 36 theme protein 1001:1007 arg1 SGP-2 955:959 SGP-2 955:959 SGP-2 955:959 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 36 theme protein 1001:1007 arg1 SP-40,40 942:949 SP-40,40 942:949 SP-40,40 942:949 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 37 theme seminal 854:860 arg1 plasma 862:867 human seminal plasma 848:867 human seminal plasma at levels comparable to those in serum 848:906 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 8 38 theme reproductive 1482:1493 arg1 systems 1495:1501 the reproductive systems 1478:1501 the reproductive systems 1478:1501 The precise role of SP-40,40 is not known in either blood or semen but the present findings document an intriguing link between the immune and the reproductive systems. 2721499 1 39 theme complement-associated 193:213 arg1 SP-40,40 224:231 SP-40,40 224:231 SP-40,40 224:231 The cDNA sequence encoding the human complement-associated protein, SP-40,40, is reported. 2721499 1 39 theme complement-associated 193:213 arg1 protein 215:221 the human complement-associated protein 183:221 the human complement-associated protein 183:221 The cDNA sequence encoding the human complement-associated protein, SP-40,40, is reported. 2721499 7 40 theme short 1186:1190 arg1 motif 1212:1216 The short cysteine-containing motif 1182:1216 The short cysteine-containing motif 1182:1216 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 8 41 theme SP-40,40 1355:1362 arg1 role 1347:1350 The precise role 1335:1350 The precise role of SP-40,40 1335:1362 The precise role of SP-40,40 is not known in either blood or semen but the present findings document an intriguing link between the immune and the reproductive systems. 2721499 5 42 theme human 848:852 arg1 plasma 862:867 human seminal plasma 848:867 human seminal plasma at levels comparable to those in serum 848:906 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 2 43 theme biosynthetic 372:383 arg1 protein 395:401 a biosynthetic precursor protein 370:401 a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond 370:482 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 0 44 theme complement 119:128 arg1 systems 147:153 the complement and reproductive systems 115:153 systems 147:153 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 3 45 theme N-linked 570:577 arg1 sites 593:597 six N-linked glycosylation sites 566:597 six N-linked glycosylation sites distributed equally between the two chains of the structure 566:657 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 0 46 theme human 53:57 arg1 protein 81:87 the novel, human complement-associated protein 42:87 protein 81:87 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 0 46 theme human 53:57 arg1 SP-40,40 90:97 SP-40,40 90:97 SP-40,40 90:97 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 4 47 theme secreted 781:788 arg1 product 790:796 the major secreted product 771:796 the major secreted product of Sertoli cells 771:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 47 theme secreted 781:788 arg1 glycoprotein-2 739:752 a rat sulphated glycoprotein-2 723:752 a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 723:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 6 48 theme acids 1033:1037 arg1 sequence 1012:1019 A sequence 1010:1019 A sequence of 23 amino acids within the beta-chain of SP-40,40 1010:1071 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 4 49 theme precursor 689:697 arg1 sequence 664:671 The sequence 660:671 The sequence of the SP-40,40 precursor 660:697 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 0 50 theme novel 46:50 arg1 protein 81:87 the novel, human complement-associated protein 42:87 protein 81:87 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 0 50 theme novel 46:50 arg1 SP-40,40 90:97 SP-40,40 90:97 SP-40,40 90:97 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 0 51 theme reproductive 134:145 arg1 systems 147:153 the complement and reproductive systems 115:153 systems 147:153 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 3 52 theme structure 649:657 arg1 chains 635:640 the two chains 627:640 the two chains of the structure 627:657 The precursor is preceded by a signal sequence for vectorial export and contains six N-linked glycosylation sites distributed equally between the two chains of the structure. 2721499 4 53 theme major 775:779 arg1 product 790:796 the major secreted product 771:796 the major secreted product of Sertoli cells 771:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 4 53 theme major 775:779 arg1 glycoprotein-2 739:752 a rat sulphated glycoprotein-2 723:752 a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells 723:813 The sequence of the SP-40,40 precursor bears a 77% identity to a rat sulphated glycoprotein-2 (SGP-2) which is the major secreted product of Sertoli cells. 2721499 7 54 theme only 1234:1237 arg1 evidence 1239:1246 the only evidence 1230:1246 the only evidence of a possible vestigial relationship between SP-40,40 and other complement components 1230:1332 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 7 55 theme cysteine-containing 1192:1210 arg1 motif 1212:1216 The short cysteine-containing motif 1182:1216 The short cysteine-containing motif 1182:1216 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 8 56 theme present 1410:1416 arg1 findings 1418:1425 the present findings 1406:1425 the present findings 1406:1425 The precise role of SP-40,40 is not known in either blood or semen but the present findings document an intriguing link between the immune and the reproductive systems. 2721499 7 57 theme complement 1312:1321 arg1 components 1323:1332 other complement components 1306:1332 other complement components 1306:1332 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 5 58 theme serum 965:969 arg1 forms 983:987 serum and seminal forms 965:987 forms 983:987 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 58 theme serum 965:969 arg1 SGP-2 955:959 SGP-2 955:959 SGP-2 955:959 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 5 58 theme serum 965:969 arg1 SP-40,40 942:949 SP-40,40 942:949 SP-40,40 942:949 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 6 59 theme located 1130:1136 arg1 segments 1121:1128 corresponding segments 1107:1128 corresponding segments located within complement components C7, C8 and C9 1107:1179 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 8 60 theme intriguing 1439:1448 arg1 link 1450:1453 an intriguing link 1436:1453 an intriguing link between the immune and the reproductive systems 1436:1501 The precise role of SP-40,40 is not known in either blood or semen but the present findings document an intriguing link between the immune and the reproductive systems. 2721499 2 61 theme reading 301:307 arg1 frame 309:313 a single open reading frame 287:313 a single open reading frame on the same mRNA molecule 287:339 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 2 62 theme bond 479:482 arg1 proteolysis 443:453 the proteolysis 439:453 the proteolysis of at least one peptide bond 439:482 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 5 63 theme same 996:999 arg1 protein 1001:1007 the same protein 992:1007 the same protein 992:1007 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 0 64 dep cloning 10:16 arg1 link 102:105 a link 100:105 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 0:154 Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. 2721499 1 65 theme human 187:191 arg1 SP-40,40 224:231 SP-40,40 224:231 SP-40,40 224:231 The cDNA sequence encoding the human complement-associated protein, SP-40,40, is reported. 2721499 1 65 theme human 187:191 arg1 protein 215:221 the human complement-associated protein 183:221 the human complement-associated protein 183:221 The cDNA sequence encoding the human complement-associated protein, SP-40,40, is reported. 2721499 5 66 theme SP-40,40 832:839 arg1 presence 820:827 The presence 816:827 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum 816:906 The presence of SP-40,40 within human seminal plasma at levels comparable to those in serum was demonstrated, indicating that SP-40,40 and SGP-2 are serum and seminal forms of the same protein. 2721499 6 67 theme significant 1083:1093 arg1 homology 1095:1102 significant homology 1083:1102 significant homology to corresponding segments located within complement components C7, C8 and C9 1083:1179 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2721499 2 68 theme peptide 471:477 arg1 bond 479:482 at least one peptide bond 458:482 at least one peptide bond 458:482 The two chains of SP-40,40 are coded in a single open reading frame on the same mRNA molecule, indicating the existence of a biosynthetic precursor protein which matures post-synthetically by the proteolysis of at least one peptide bond. 2721499 7 69 theme other 1306:1310 arg1 components 1323:1332 other complement components 1306:1332 other complement components 1306:1332 The short cysteine-containing motif represented the only evidence of a possible vestigial relationship between SP-40,40 and other complement components. 2721499 6 70 theme amino 1027:1031 arg1 acids 1033:1037 23 amino acids 1024:1037 23 amino acids 1024:1037 A sequence of 23 amino acids within the beta-chain of SP-40,40 exhibited significant homology to corresponding segments located within complement components C7, C8 and C9. 2361960 6 0 theme rhIL-5 883:888 arg1 structure 870:878 primary structure 862:878 primary structure of rhIL-5 862:888 The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate. 2361960 5 1 gly glycosylated 822:833 arg1 Asn-28 810:815 Asn-28 810:815 Asn-28 810:815 In addition, we concluded that Thr-3 and Asn-28 were glycosylated. 2361960 5 1 gly glycosylated 822:833 arg1 Thr-3 800:804 Thr-3 800:804 Thr-3 800:804 In addition, we concluded that Thr-3 and Asn-28 were glycosylated. 2361960 0 2 theme ovary 73:77 arg1 cells 79:83 Chinese hamster ovary cells 57:83 Chinese hamster ovary cells 57:83 Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells. 2361960 2 3 theme peptide 271:277 arg1 fragment 279:286 Each peptide fragment 266:286 Each peptide fragment generated by Achromobacter protease I (API) digestion 266:340 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 3 4 theme cDNA 527:530 arg1 sequence 532:539 human IL-5 cDNA sequence 516:539 human IL-5 cDNA sequence 516:539 After digestion with API, we could identify all the peptides which were expected from human IL-5 cDNA sequence. 2361960 1 5 theme disulfide 237:245 arg1 structure 255:263 disulfide bonding structure 237:263 disulfide bonding structure 237:263 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 6 6 theme primary 862:868 arg1 structure 870:878 primary structure 862:878 primary structure of rhIL-5 862:888 The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate. 2361960 3 7 theme human 516:520 arg1 sequence 532:539 human IL-5 cDNA sequence 516:539 human IL-5 cDNA sequence 516:539 After digestion with API, we could identify all the peptides which were expected from human IL-5 cDNA sequence. 2361960 1 8 theme bonding 247:253 arg1 structure 255:263 disulfide bonding structure 237:263 disulfide bonding structure 237:263 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 4 9 from analyses 546:553 arg1 molecule 587:594 rhIL-5 molecule 580:594 rhIL-5 molecule 580:594 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 6 10 from difference 953:962 arg1 content 971:977 the content 967:977 the content of carbohydrate 967:993 The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate. 2361960 1 11 gly glycosylation 212:224 arg2 sites 226:230 glycosylation sites 212:230 glycosylation sites 212:230 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 4 12 theme Cys-44 750:755 arg1 pairs 741:745 two pairs 737:745 two pairs of Cys-44 and Cys-86 737:766 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 2 13 theme amino 376:380 arg1 analysis 387:394 amino acid analysis 376:394 amino acid analysis 376:394 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 2 14 theme protease 315:322 arg1 digestion 332:340 Achromobacter protease I (API) digestion 301:340 Achromobacter protease I (API) digestion 301:340 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 4 15 from content 569:575 arg1 molecule 587:594 rhIL-5 molecule 580:594 rhIL-5 molecule 580:594 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 16 from molecule 587:594 arg1 peptide 621:627 disulfide-containing peptide 600:627 disulfide-containing peptide obtained from API digestion 600:655 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 16 from molecule 587:594 arg1 analyses 546:553 The analyses 542:553 The analyses of sulfhydryl content in rhIL-5 molecule 542:594 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 2 17 theme Achromobacter 301:313 arg1 digestion 332:340 Achromobacter protease I (API) digestion 301:340 Achromobacter protease I (API) digestion 301:340 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 4 18 theme API 643:645 arg1 digestion 647:655 API digestion 643:655 API digestion 643:655 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 19 theme antiparallel 708:719 arg1 dimer 721:725 an antiparallel dimer 705:725 an antiparallel dimer linked by two pairs of Cys-44 and Cys-86 705:766 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 0 20 theme human 25:29 arg1 interleukin 31:41 recombinant human interleukin 5 13:43 recombinant human interleukin 5 13:43 Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells. 2361960 1 21 theme complete 90:97 arg1 map 107:109 The complete peptide map 86:109 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) 86:162 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 4 22 from peptide 621:627 arg1 molecule 587:594 rhIL-5 molecule 580:594 rhIL-5 molecule 580:594 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 2 23 theme amino 400:404 arg1 analysis 420:427 amino acid sequence analysis 400:427 amino acid sequence analysis 400:427 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 2 24 theme acid 406:409 arg1 analysis 420:427 amino acid sequence analysis 400:427 amino acid sequence analysis 400:427 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 3 25 with digestion 436:444 arg1 API 451:453 API 451:453 API 451:453 After digestion with API, we could identify all the peptides which were expected from human IL-5 cDNA sequence. 2361960 1 26 theme peptide 99:105 arg1 map 107:109 The complete peptide map 86:109 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) 86:162 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 4 27 theme active 672:677 arg1 form 679:682 active form 672:682 active form of rhIL-5 672:692 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 0 28 theme interleukin 31:41 arg1 Structure 0:8 Structure 0:8 Structure of recombinant human interleukin 5 0:43 Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells. 2361960 6 29 theme observed 916:923 arg1 heterogeneity 925:937 observed heterogeneity 916:937 observed heterogeneity 916:937 The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate. 2361960 2 30 theme sequence 411:418 arg1 analysis 420:427 amino acid sequence analysis 400:427 amino acid sequence analysis 400:427 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 1 31 theme primary 193:199 arg1 structure 201:209 its primary structure 189:209 its primary structure 189:209 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 2 32 theme API 327:329 arg1 digestion 332:340 Achromobacter protease I (API) digestion 301:340 Achromobacter protease I (API) digestion 301:340 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 4 33 theme content 569:575 arg1 peptide 621:627 disulfide-containing peptide 600:627 disulfide-containing peptide obtained from API digestion 600:655 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 33 theme content 569:575 arg1 analyses 546:553 The analyses 542:553 The analyses of sulfhydryl content in rhIL-5 molecule 542:594 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 34 theme sulfhydryl 558:567 arg1 content 569:575 sulfhydryl content 558:575 sulfhydryl content in rhIL-5 molecule 558:594 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 35 theme rhIL-5 687:692 arg1 form 679:682 active form 672:682 active form of rhIL-5 672:692 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 4 36 theme Cys-86 761:766 arg1 pairs 741:745 two pairs 737:745 two pairs of Cys-44 and Cys-86 737:766 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 6 37 theme carbohydrate 982:993 arg1 content 971:977 the content 967:977 the content of carbohydrate 967:993 The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate. 2361960 1 38 theme purified 114:121 arg1 interleukin 141:151 human interleukin 5 135:153 purified recombinant human interleukin 5 (rhIL-5) 114:162 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 1 38 theme purified 114:121 arg1 rhIL-5 156:161 rhIL-5 156:161 rhIL-5 156:161 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 3 39 theme IL-5 522:525 arg1 sequence 532:539 human IL-5 cDNA sequence 516:539 human IL-5 cDNA sequence 516:539 After digestion with API, we could identify all the peptides which were expected from human IL-5 cDNA sequence. 2361960 1 40 theme recombinant 123:133 arg1 interleukin 141:151 human interleukin 5 135:153 purified recombinant human interleukin 5 (rhIL-5) 114:162 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 1 40 theme recombinant 123:133 arg1 rhIL-5 156:161 rhIL-5 156:161 rhIL-5 156:161 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 2 41 theme I 324:324 arg1 digestion 332:340 Achromobacter protease I (API) digestion 301:340 Achromobacter protease I (API) digestion 301:340 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 0 42 theme hamster 65:71 arg1 cells 79:83 Chinese hamster ovary cells 57:83 Chinese hamster ovary cells 57:83 Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells. 2361960 2 43 theme acid 382:385 arg1 analysis 387:394 amino acid analysis 376:394 amino acid analysis 376:394 Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis. 2361960 4 44 theme disulfide-containing 600:619 arg1 peptide 621:627 disulfide-containing peptide 600:627 disulfide-containing peptide obtained from API digestion 600:655 The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86. 2361960 1 45 theme human 135:139 arg1 interleukin 141:151 human interleukin 5 135:153 purified recombinant human interleukin 5 (rhIL-5) 114:162 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 1 45 theme human 135:139 arg1 rhIL-5 156:161 rhIL-5 156:161 rhIL-5 156:161 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 1 46 theme glycosylation 212:224 arg1 sites 226:230 glycosylation sites 212:230 glycosylation sites 212:230 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 2361960 0 47 theme Chinese 57:63 arg1 cells 79:83 Chinese hamster ovary cells 57:83 Chinese hamster ovary cells 57:83 Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells. 2361960 0 48 theme recombinant 13:23 arg1 interleukin 31:41 recombinant human interleukin 5 13:43 recombinant human interleukin 5 13:43 Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells. 2361960 1 49 theme interleukin 141:151 arg1 map 107:109 The complete peptide map 86:109 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) 86:162 The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. 1740433 1 0 theme non-acute 234:242 arg1 lipoprotein 263:273 non-acute phase high density lipoprotein 234:273 non-acute phase high density lipoprotein 234:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 8 1 link N-linked 1169:1176 arg1 site 1192:1195 the only potential N-linked glycosylation site 1150:1195 the only potential N-linked glycosylation site in the molecule 1150:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 1 link N-linked 1169:1176 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 3 2 theme SAA 527:529 arg1 superfamily 531:541 the SAA superfamily 523:541 the SAA superfamily 523:541 C-SAA was partially sequenced, and immunochemical analyses indicated that it constitutes a distinct subclass of apolipoproteins within the SAA superfamily. 1740433 6 3 theme mature 742:747 arg1 molecule 749:756 The 112-residue mature molecule 726:756 The 112-residue mature molecule 726:756 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 6 3 theme mature 742:747 arg1 residues 763:770 8 residues 761:770 8 residues longer than human A-SAA 761:794 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 1 4 theme high 250:253 arg1 lipoprotein 263:273 non-acute phase high density lipoprotein 234:273 non-acute phase high density lipoprotein 234:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 8 5 theme C-SAA 1078:1082 arg1 octapeptide 1084:1094 The C-SAA octapeptide 1074:1094 The C-SAA octapeptide 1074:1094 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 6 6 theme size 801:804 arg1 due 820:822 due 820:822 due 820:822 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 6 6 theme size 801:804 arg1 difference 806:815 the size difference 797:815 the size difference 797:815 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 8 7 theme tripeptide 1138:1147 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 7 theme tripeptide 1138:1147 arg1 residues 1120:1127 the first two residues 1106:1127 the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule 1106:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 10 8 theme A-SAA 1438:1442 arg1 mRNA 1444:1447 A-SAA mRNA 1438:1447 A-SAA mRNA 1438:1447 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 5 9 theme pre-C-SAA 639:647 arg1 molecule 649:656 a pre-C-SAA molecule 637:656 a pre-C-SAA molecule of 130 residues from which an 18-residue leader peptide is cleaved 637:723 The clone predicts a pre-C-SAA molecule of 130 residues from which an 18-residue leader peptide is cleaved. 1740433 3 10 theme apolipoproteins 500:514 arg1 subclass 488:495 a distinct subclass 477:495 a distinct subclass of apolipoproteins within the SAA superfamily 477:541 C-SAA was partially sequenced, and immunochemical analyses indicated that it constitutes a distinct subclass of apolipoproteins within the SAA superfamily. 1740433 8 11 theme first 1110:1114 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 11 theme first 1110:1114 arg1 residues 1120:1127 the first two residues 1106:1127 the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule 1106:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 4 12 theme human 583:587 arg1 library 595:601 a human liver library 581:601 a human liver library 581:601 A C-SAA cDNA clone was isolated from a human liver library and sequenced. 1740433 8 13 theme glycosylation 1178:1190 arg1 site 1192:1195 the only potential N-linked glycosylation site 1150:1195 the only potential N-linked glycosylation site in the molecule 1150:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 13 theme glycosylation 1178:1190 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 10 14 theme A-SAA 1633:1637 arg1 mRNA 1639:1642 A-SAA mRNA 1633:1642 A-SAA mRNA observed 1633:1651 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 9 15 theme C-SAA 1346:1350 arg1 kDa 1338:1340 14 and 19 kDa 1328:1340 14 and 19 kDa 1328:1340 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 9 15 theme C-SAA 1346:1350 arg1 classes 1319:1325 two size classes 1310:1325 two size classes 1310:1325 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 7 16 located found 1003:1007 arg1 positions 1020:1028 similar positions 1012:1028 similar positions in the A-SAAs of a number of other species 1012:1071 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 7 16 located found 1003:1007 arg2 octapeptides 963:974 octapeptides 963:974 octapeptides of similar composition 963:997 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 5 17 theme 18-residue 688:697 arg1 peptide 706:712 an 18-residue leader peptide 685:712 an 18-residue leader peptide 685:712 The clone predicts a pre-C-SAA molecule of 130 residues from which an 18-residue leader peptide is cleaved. 1740433 7 18 theme other 1059:1063 arg1 species 1065:1071 other species 1059:1071 other species 1059:1071 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 1 19 theme serum 144:148 arg1 SAA 169:171 SAA 169:171 SAA 169:171 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 19 theme serum 144:148 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 19 theme serum 144:148 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 0 20 theme superfamily 63:73 arg1 members 24:30 novel members 18:30 novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein 18:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 1 21 theme A 158:158 arg1 SAA 169:171 SAA 169:171 SAA 169:171 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 21 theme A 158:158 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 21 theme A 158:158 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 6 22 located found 898:902 arg2 positions 866:874 positions 70 and 77 866:884 positions 866:874 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 6 22 located found 898:902 arg2 77 883:884 77 883:884 77 883:884 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 6 22 located found 898:902 arg1 region 925:930 the corresponding region 907:930 the corresponding region of human A-SAA 907:945 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 0 23 theme constitutive 78:89 arg1 apolipoproteins 91:105 constitutive apolipoproteins 78:105 constitutive apolipoproteins of high density lipoprotein 78:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 10 24 contain contains 1385:1392 arg1 liver 1379:1383 Human acute phase liver 1361:1383 Human acute phase liver 1361:1383 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 10 24 contain contains 1385:1392 arg2 mRNA 1407:1410 little C-SAA mRNA 1394:1410 little C-SAA mRNA relative to the levels of A-SAA mRNA 1394:1447 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 10 25 theme cells 1490:1494 arg1 treatment 1458:1466 the treatment 1454:1466 the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium 1454:1527 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 10 26 theme PLC/PRF/5 1471:1479 arg1 cells 1490:1494 PLC/PRF/5 hepatoma cells 1471:1494 PLC/PRF/5 hepatoma cells 1471:1494 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 1 27 theme density 255:261 arg1 lipoprotein 263:273 non-acute phase high density lipoprotein 234:273 non-acute phase high density lipoprotein 234:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 2 28 theme " 373:373 arg1 A-SAA 380:384 A-SAA 380:384 A-SAA 380:384 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 2 28 theme " 373:373 arg1 SAA 375:377 "acute phase" SAA 361:377 "acute phase" SAA (A-SAA) 361:385 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 0 29 theme novel 18:22 arg1 members 24:30 novel members 18:30 novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein 18:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 10 30 theme monocyte-conditioned 1501:1520 arg1 medium 1522:1527 monocyte-conditioned medium 1501:1527 monocyte-conditioned medium 1501:1527 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 9 31 gly glycosylated 1273:1284 arg1 % 1252:1252 approximately 50% 1236:1252 approximately 50% of these sites 1236:1267 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 9 31 gly glycosylated 1273:1284 arg1 sites 1263:1267 these sites 1257:1267 these sites 1257:1267 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 2 32 theme acute 362:366 arg1 A-SAA 380:384 A-SAA 380:384 A-SAA 380:384 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 2 32 theme acute 362:366 arg1 SAA 375:377 "acute phase" SAA 361:377 "acute phase" SAA (A-SAA) 361:385 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 7 33 theme number 1049:1054 arg1 A-SAAs 1037:1042 the A-SAAs 1033:1042 the A-SAAs of a number of other species 1033:1071 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 6 34 theme human 935:939 arg1 A-SAA 941:945 human A-SAA 935:945 human A-SAA 935:945 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 7 35 from positions 1020:1028 arg1 A-SAAs 1037:1042 the A-SAAs 1033:1042 the A-SAAs of a number of other species 1033:1071 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 10 36 theme phase 1373:1377 arg1 liver 1379:1383 Human acute phase liver 1361:1383 Human acute phase liver 1361:1383 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 0 37 theme A 53:53 arg1 superfamily 63:73 the serum amyloid A protein superfamily 35:73 the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein 35:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 10 38 theme mRNA 1551:1554 arg1 concentrations 1556:1569 C-SAA mRNA concentrations 1545:1569 C-SAA mRNA concentrations 1545:1569 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 0 39 theme serum 39:43 arg1 superfamily 63:73 the serum amyloid A protein superfamily 35:73 the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein 35:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 10 40 theme Human 1361:1365 arg1 liver 1379:1383 Human acute phase liver 1361:1383 Human acute phase liver 1361:1383 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 4 41 theme C-SAA 546:550 arg1 clone 557:561 A C-SAA cDNA clone 544:561 A C-SAA cDNA clone 544:561 A C-SAA cDNA clone was isolated from a human liver library and sequenced. 1740433 10 42 theme C-SAA 1401:1405 arg1 mRNA 1407:1410 little C-SAA mRNA 1394:1410 little C-SAA mRNA relative to the levels of A-SAA mRNA 1394:1447 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 10 43 theme C-SAA 1545:1549 arg1 concentrations 1556:1569 C-SAA mRNA concentrations 1545:1569 C-SAA mRNA concentrations 1545:1569 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 1 44 theme apolipoprotein 206:219 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 44 theme apolipoprotein 206:219 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 10 45 with treatment 1458:1466 arg1 medium 1522:1527 monocyte-conditioned medium 1501:1527 monocyte-conditioned medium 1501:1527 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 7 46 theme similar 1012:1018 arg1 positions 1020:1028 similar positions 1012:1028 similar positions in the A-SAAs of a number of other species 1012:1071 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 3 47 theme distinct 479:486 arg1 subclass 488:495 a distinct subclass 477:495 a distinct subclass of apolipoproteins within the SAA superfamily 477:541 C-SAA was partially sequenced, and immunochemical analyses indicated that it constitutes a distinct subclass of apolipoproteins within the SAA superfamily. 1740433 1 48 theme phase 244:248 arg1 lipoprotein 263:273 non-acute phase high density lipoprotein 234:273 non-acute phase high density lipoprotein 234:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 10 49 theme relative 1412:1419 arg1 mRNA 1407:1410 little C-SAA mRNA 1394:1410 little C-SAA mRNA relative to the levels of A-SAA mRNA 1394:1447 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 2 50 theme " 324:324 arg1 C-SAA 331:335 C-SAA 331:335 C-SAA 331:335 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 2 50 theme " 324:324 arg1 SAA 326:328 constitutive" SAA 312:328 constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA) 312:385 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 6 51 theme 112-residue 730:740 arg1 molecule 749:756 The 112-residue mature molecule 726:756 The 112-residue mature molecule 726:756 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 6 51 theme 112-residue 730:740 arg1 residues 763:770 8 residues 761:770 8 residues longer than human A-SAA 761:794 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 9 52 theme size 1314:1317 arg1 kDa 1338:1340 14 and 19 kDa 1328:1340 14 and 19 kDa 1328:1340 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 9 52 theme size 1314:1317 arg1 classes 1319:1325 two size classes 1310:1325 two size classes 1310:1325 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 7 53 theme similar 979:985 arg1 composition 987:997 similar composition 979:997 similar composition 979:997 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 4 54 theme liver 589:593 arg1 library 595:601 a human liver library 581:601 a human liver library 581:601 A C-SAA cDNA clone was isolated from a human liver library and sequenced. 1740433 10 55 theme mRNA 1444:1447 arg1 levels 1428:1433 the levels 1424:1433 the levels of A-SAA mRNA 1424:1447 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 7 56 theme composition 987:997 arg1 octapeptides 963:974 octapeptides 963:974 octapeptides of similar composition 963:997 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 8 57 theme NSS 1134:1136 arg1 site 1192:1195 the only potential N-linked glycosylation site 1150:1195 the only potential N-linked glycosylation site in the molecule 1150:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 57 theme NSS 1134:1136 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 0 58 theme lipoprotein 123:133 arg1 apolipoproteins 91:105 constitutive apolipoproteins 78:105 constitutive apolipoproteins of high density lipoprotein 78:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 10 59 theme massive 1612:1618 arg1 induction 1620:1628 the massive induction 1608:1628 the massive induction of A-SAA mRNA observed 1608:1651 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 2 60 theme constitutive 312:323 arg1 C-SAA 331:335 C-SAA 331:335 C-SAA 331:335 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 2 60 theme constitutive 312:323 arg1 SAA 326:328 constitutive" SAA 312:328 constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA) 312:385 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 11 61 theme acute 1685:1689 arg1 C-SAA 1654:1658 C-SAA 1654:1658 C-SAA 1654:1658 C-SAA is therefore not a major acute phase reactant. 1740433 11 61 theme acute 1685:1689 arg1 reactant 1697:1704 a major acute phase reactant 1677:1704 a major acute phase reactant 1677:1704 C-SAA is therefore not a major acute phase reactant. 1740433 10 62 theme mRNA 1639:1642 arg1 induction 1620:1628 the massive induction 1608:1628 the massive induction of A-SAA mRNA observed 1608:1651 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 6 63 theme octapeptide 846:856 arg1 presence 831:838 the presence 827:838 the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA 827:945 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 8 64 theme N-linked 1169:1176 arg1 site 1192:1195 the only potential N-linked glycosylation site 1150:1195 the only potential N-linked glycosylation site in the molecule 1150:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 64 theme N-linked 1169:1176 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 10 65 dep induction 1620:1628 arg1 contrast 1596:1603 contrast 1596:1603 contrast 1596:1603 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 0 66 theme protein 55:61 arg1 superfamily 63:73 the serum amyloid A protein superfamily 35:73 the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein 35:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 1 67 theme novel 138:142 arg1 SAA 169:171 SAA 169:171 SAA 169:171 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 67 theme novel 138:142 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 67 theme novel 138:142 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 6 68 theme corresponding 911:923 arg1 region 925:930 the corresponding region 907:930 the corresponding region of human A-SAA 907:945 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 5 69 theme residues 665:672 arg1 molecule 649:656 a pre-C-SAA molecule 637:656 a pre-C-SAA molecule of 130 residues from which an 18-residue leader peptide is cleaved 637:723 The clone predicts a pre-C-SAA molecule of 130 residues from which an 18-residue leader peptide is cleaved. 1740433 5 70 theme leader 699:704 arg1 peptide 706:712 an 18-residue leader peptide 685:712 an 18-residue leader peptide 685:712 The clone predicts a pre-C-SAA molecule of 130 residues from which an 18-residue leader peptide is cleaved. 1740433 8 71 gly glycosylation 1178:1190 arg2 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 71 gly glycosylation 1178:1190 arg2 site 1192:1195 the only potential N-linked glycosylation site 1150:1195 the only potential N-linked glycosylation site in the molecule 1150:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 1 72 theme amyloid 150:156 arg1 SAA 169:171 SAA 169:171 SAA 169:171 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 72 theme amyloid 150:156 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 72 theme amyloid 150:156 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 0 73 theme density 115:121 arg1 lipoprotein 123:133 high density lipoprotein 110:133 high density lipoprotein 110:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 8 74 from site 1192:1195 arg1 molecule 1204:1211 the molecule 1200:1211 the molecule 1200:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 2 75 theme novel 281:285 arg1 SAA 287:289 This novel SAA 276:289 This novel SAA 276:289 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 0 76 theme high 110:113 arg1 lipoprotein 123:133 high density lipoprotein 110:133 high density lipoprotein 110:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 10 77 theme hepatoma 1481:1488 arg1 cells 1490:1494 PLC/PRF/5 hepatoma cells 1471:1494 PLC/PRF/5 hepatoma cells 1471:1494 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 7 78 theme species 1065:1071 arg1 number 1049:1054 a number 1047:1054 a number of other species 1047:1071 Paradoxically, octapeptides of similar composition are found at similar positions in the A-SAAs of a number of other species. 1740433 0 79 theme members 24:30 arg1 Identification 0:13 Identification 0:13 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 0:134 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 1 80 theme lipoprotein 263:273 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 80 theme lipoprotein 263:273 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 2 81 theme phase 368:372 arg1 A-SAA 380:384 A-SAA 380:384 A-SAA 380:384 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 2 81 theme phase 368:372 arg1 SAA 375:377 "acute phase" SAA 361:377 "acute phase" SAA (A-SAA) 361:385 This novel SAA has been designated "constitutive" SAA (C-SAA) to distinguish it from "acute phase" SAA (A-SAA). 1740433 8 82 theme potential 1159:1167 arg1 site 1192:1195 the only potential N-linked glycosylation site 1150:1195 the only potential N-linked glycosylation site in the molecule 1150:1211 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 8 82 theme potential 1159:1167 arg1 tripeptide 1138:1147 a NSS tripeptide 1132:1147 a NSS tripeptide 1132:1147 The C-SAA octapeptide specifies the first two residues of a NSS tripeptide, the only potential N-linked glycosylation site in the molecule. 1740433 6 83 theme A-SAA 941:945 arg1 region 925:930 the corresponding region 907:930 the corresponding region of human A-SAA 907:945 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 11 84 theme phase 1691:1695 arg1 C-SAA 1654:1658 C-SAA 1654:1658 C-SAA 1654:1658 C-SAA is therefore not a major acute phase reactant. 1740433 11 84 theme phase 1691:1695 arg1 reactant 1697:1704 a major acute phase reactant 1677:1704 a major acute phase reactant 1677:1704 C-SAA is therefore not a major acute phase reactant. 1740433 0 85 theme amyloid 45:51 arg1 superfamily 63:73 the serum amyloid A protein superfamily 35:73 the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein 35:133 Identification of novel members of the serum amyloid A protein superfamily as constitutive apolipoproteins of high density lipoprotein. 1740433 6 86 theme human 784:788 arg1 A-SAA 790:794 human A-SAA 784:794 human A-SAA 784:794 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 10 87 theme acute 1367:1371 arg1 liver 1379:1383 Human acute phase liver 1361:1383 Human acute phase liver 1361:1383 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 4 88 attach isolated 567:574 arg1 library 595:601 a human liver library 581:601 a human liver library 581:601 A C-SAA cDNA clone was isolated from a human liver library and sequenced. 1740433 4 88 attach isolated 567:574 arg2 clone 557:561 A C-SAA cDNA clone 544:561 A C-SAA cDNA clone 544:561 A C-SAA cDNA clone was isolated from a human liver library and sequenced. 1740433 3 89 theme immunochemical 423:436 arg1 analyses 438:445 immunochemical analyses 423:445 immunochemical analyses 423:445 C-SAA was partially sequenced, and immunochemical analyses indicated that it constitutes a distinct subclass of apolipoproteins within the SAA superfamily. 1740433 9 90 theme sites 1263:1267 arg1 % 1252:1252 approximately 50% 1236:1252 approximately 50% of these sites 1236:1267 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 9 90 theme sites 1263:1267 arg1 sites 1263:1267 these sites 1257:1267 these sites 1257:1267 Studies indicate that approximately 50% of these sites are glycosylated, thereby giving rise to two size classes, 14 and 19 kDa, of C-SAA in vivo. 1740433 6 91 theme longer 772:777 arg1 molecule 749:756 The 112-residue mature molecule 726:756 The 112-residue mature molecule 726:756 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 6 91 theme longer 772:777 arg1 residues 763:770 8 residues 761:770 8 residues longer than human A-SAA 761:794 The 112-residue mature molecule is 8 residues longer than human A-SAA; the size difference is due to the presence of an octapeptide between positions 70 and 77 that is not found in the corresponding region of human A-SAA. 1740433 4 92 theme cDNA 552:555 arg1 clone 557:561 A C-SAA cDNA clone 544:561 A C-SAA cDNA clone 544:561 A C-SAA cDNA clone was isolated from a human liver library and sequenced. 1740433 11 93 theme major 1679:1683 arg1 C-SAA 1654:1658 C-SAA 1654:1658 C-SAA 1654:1658 C-SAA is therefore not a major acute phase reactant. 1740433 11 93 theme major 1679:1683 arg1 reactant 1697:1704 a major acute phase reactant 1677:1704 a major acute phase reactant 1677:1704 C-SAA is therefore not a major acute phase reactant. 1740433 1 94 theme normal 199:204 arg1 protein 160:166 A novel serum amyloid A protein 136:166 A novel serum amyloid A protein (SAA) 136:172 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 1 94 theme normal 199:204 arg1 component 221:229 a normal apolipoprotein component 197:229 a normal apolipoprotein component of non-acute phase high density lipoprotein 197:273 A novel serum amyloid A protein (SAA) has been identified as a normal apolipoprotein component of non-acute phase high density lipoprotein. 1740433 10 95 theme little 1394:1399 arg1 mRNA 1407:1410 little C-SAA mRNA 1394:1410 little C-SAA mRNA relative to the levels of A-SAA mRNA 1394:1447 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 1740433 10 96 theme detectable 1574:1583 arg1 levels 1585:1590 detectable levels 1574:1590 detectable levels 1574:1590 Human acute phase liver contains little C-SAA mRNA relative to the levels of A-SAA mRNA, and the treatment of PLC/PRF/5 hepatoma cells with monocyte-conditioned medium does not induce C-SAA mRNA concentrations to detectable levels, in contrast to the massive induction of A-SAA mRNA observed. 17065148 1 0 theme hydrophobicity 305:318 arg1 predictions 320:330 their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions 216:330 their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 216:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 1 theme TRPM8 1081:1085 arg1 protein 1087:1093 TRPM8 protein 1081:1093 TRPM8 protein 1081:1093 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 3 2 theme single 641:646 arg1 mutation 654:661 a single point mutation 639:661 a single point mutation within the coiled coil region 639:691 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 7 3 theme protein 1087:1093 arg1 maturation 1067:1076 the maturation 1063:1076 the maturation of TRPM8 protein 1063:1093 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 1 4 from members 152:158 arg1 distinct 127:134 distinct 127:134 distinct 127:134 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 4 from members 152:158 arg1 regard 206:211 regard 206:211 regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 206:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 5 theme amino 242:246 arg1 size 230:233 their overall size 216:233 their overall size (>1000 amino acids) 216:253 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 5 theme amino 242:246 arg1 acids 248:252 >1000 amino acids 236:252 >1000 amino acids 236:252 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 3 6 theme point 648:652 arg1 mutation 654:661 a single point mutation 639:661 a single point mutation within the coiled coil region 639:691 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 3 7 theme putative 532:539 arg1 region 553:558 the putative coiled coil region 528:558 the putative coiled coil region within the intracellular C terminus 528:594 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 3 8 theme coil 681:684 arg1 region 686:691 the coiled coil region 670:691 the coiled coil region 670:691 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 1 9 theme TRPM 62:65 arg1 potential 87:95 transient receptor potential melastatin-like 68:111 transient receptor potential melastatin-like 68:111 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 9 theme TRPM 62:65 arg1 channels 114:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 10 from trafficking 1034:1044 arg1 assembly 1020:1027 separate assembly 1011:1027 separate assembly from trafficking 1011:1044 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 7 11 theme relevant 1127:1134 arg1 site 1159:1162 the relevant N-linked glycosylation site 1123:1162 the relevant N-linked glycosylation site 1123:1162 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 4 12 theme functional 728:737 arg1 channels 739:746 functional channels 728:746 functional channels 728:746 This mutant neither gives rise to functional channels nor do its subunits interact or form protein complexes that correspond to a multimer. 17065148 2 13 theme prominent 420:428 arg1 region 453:458 a prominent C-terminal coiled coil region 418:458 a prominent C-terminal coiled coil region 418:458 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 13 theme prominent 420:428 arg1 member 408:413 Common to each TRPM member 388:413 Common to each TRPM member 388:413 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 14 theme coil 448:451 arg1 region 453:458 a prominent C-terminal coiled coil region 418:458 a prominent C-terminal coiled coil region 418:458 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 14 theme coil 448:451 arg1 member 408:413 Common to each TRPM member 388:413 Common to each TRPM member 388:413 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 1 15 theme transient 68:76 arg1 potential 87:95 transient receptor potential melastatin-like 68:111 transient receptor potential melastatin-like 68:111 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 15 theme transient 68:76 arg1 channels 114:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 16 theme transient 167:175 arg1 family 196:201 the transient receptor potential family 163:201 the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 163:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 17 theme N-linked 1136:1143 arg1 site 1159:1162 the relevant N-linked glycosylation site 1123:1162 the relevant N-linked glycosylation site 1123:1162 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 3 18 theme coiled 674:679 arg1 region 686:691 the coiled coil region 670:691 the coiled coil region 670:691 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 1 19 theme receptor 78:85 arg1 potential 87:95 transient receptor potential melastatin-like 68:111 transient receptor potential melastatin-like 68:111 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 19 theme receptor 78:85 arg1 channels 114:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 20 theme receptor 177:184 arg1 family 196:201 the transient receptor potential family 163:201 the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 163:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 21 theme glycosylation 1145:1157 arg1 site 1159:1162 the relevant N-linked glycosylation site 1123:1162 the relevant N-linked glycosylation site 1123:1162 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 3 22 theme coiled 541:546 arg1 region 553:558 the putative coiled coil region 528:558 the putative coiled coil region within the intracellular C terminus 528:594 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 1 23 theme potential 186:194 arg1 family 196:201 the transient receptor potential family 163:201 the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 163:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 24 link N-linked 1136:1143 arg1 site 1159:1162 the relevant N-linked glycosylation site 1123:1162 the relevant N-linked glycosylation site 1123:1162 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 6 25 theme wild-type 907:915 arg1 currents 917:924 wild-type currents 907:924 wild-type currents 907:924 Furthermore, wild-type currents can be suppressed by expressing the membrane-attached C-terminal region of TRPM8. 17065148 1 26 theme melastatin-like 97:111 arg1 potential 87:95 transient receptor potential melastatin-like 68:111 transient receptor potential melastatin-like 68:111 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 26 theme melastatin-like 97:111 arg1 channels 114:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels 62:121 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 27 theme family 196:201 arg1 members 152:158 many other members 141:158 many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 141:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 2 28 theme TRPM 403:406 arg1 region 453:458 a prominent C-terminal coiled coil region 418:458 a prominent C-terminal coiled coil region 418:458 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 28 theme TRPM 403:406 arg1 member 408:413 Common to each TRPM member 388:413 Common to each TRPM member 388:413 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 1 29 theme lack 260:263 arg1 predictions 320:330 their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions 216:330 their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 216:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 0 30 theme cold-sensitive 32:45 arg1 channel 53:59 the cold-sensitive TRPM8 channel 28:59 the cold-sensitive TRPM8 channel 28:59 Trafficking and assembly of the cold-sensitive TRPM8 channel. 17065148 7 31 theme separate 1011:1018 arg1 assembly 1020:1027 separate assembly 1011:1027 separate assembly from trafficking 1011:1044 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 6 32 theme TRPM8 1001:1005 arg1 region 991:996 the membrane-attached C-terminal region 958:996 the membrane-attached C-terminal region of TRPM8 958:1005 Furthermore, wild-type currents can be suppressed by expressing the membrane-attached C-terminal region of TRPM8. 17065148 2 33 theme Common 388:393 arg1 region 453:458 a prominent C-terminal coiled coil region 418:458 a prominent C-terminal coiled coil region 418:458 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 33 theme Common 388:393 arg1 member 408:413 Common to each TRPM member 388:413 Common to each TRPM member 388:413 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 34 theme coiled 441:446 arg1 region 453:458 a prominent C-terminal coiled coil region 418:458 a prominent C-terminal coiled coil region 418:458 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 34 theme coiled 441:446 arg1 member 408:413 Common to each TRPM member 388:413 Common to each TRPM member 388:413 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 1 35 theme N-terminal 268:277 arg1 repeats 292:298 N-terminal ankyrin-like repeats 268:298 N-terminal ankyrin-like repeats 268:298 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 36 theme transmembrane 365:377 arg1 regions 379:385 more than six transmembrane regions 351:385 more than six transmembrane regions 351:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 6 37 theme membrane-attached 962:978 arg1 region 991:996 the membrane-attached C-terminal region 958:996 the membrane-attached C-terminal region of TRPM8 958:1005 Furthermore, wild-type currents can be suppressed by expressing the membrane-attached C-terminal region of TRPM8. 17065148 1 38 from regard 206:211 arg1 members 152:158 many other members 141:158 many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 141:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 0 39 theme channel 53:59 arg1 assembly 16:23 assembly 16:23 assembly 16:23 Trafficking and assembly of the cold-sensitive TRPM8 channel. 17065148 0 39 theme channel 53:59 arg1 Trafficking 0:10 Trafficking 0:10 Trafficking 0:10 Trafficking and assembly of the cold-sensitive TRPM8 channel. 17065148 1 40 theme ankyrin-like 279:290 arg1 repeats 292:298 N-terminal ankyrin-like repeats 268:298 N-terminal ankyrin-like repeats 268:298 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 3 41 theme coil 548:551 arg1 region 553:558 the putative coiled coil region 528:558 the putative coiled coil region within the intracellular C terminus 528:594 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 0 42 theme TRPM8 47:51 arg1 channel 53:59 the cold-sensitive TRPM8 channel 28:59 the cold-sensitive TRPM8 channel 28:59 Trafficking and assembly of the cold-sensitive TRPM8 channel. 17065148 1 43 theme repeats 292:298 arg1 size 230:233 their overall size 216:233 their overall size (>1000 amino acids) 216:253 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 43 theme repeats 292:298 arg1 lack 260:263 the lack 256:263 the lack of N-terminal ankyrin-like repeats 256:298 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 43 theme repeats 292:298 arg1 acids 248:252 >1000 amino acids 236:252 >1000 amino acids 236:252 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 43 theme repeats 292:298 arg1 hydrophobicity 305:318 hydrophobicity 305:318 hydrophobicity 305:318 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 4 44 theme protein 785:791 arg1 complexes 793:801 protein complexes 785:801 protein complexes that correspond to a multimer 785:831 This mutant neither gives rise to functional channels nor do its subunits interact or form protein complexes that correspond to a multimer. 17065148 1 45 from family 196:201 arg1 regard 206:211 regard 206:211 regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 206:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 46 theme plasma 1261:1266 arg1 membrane 1268:1275 the plasma membrane 1257:1275 the plasma membrane 1257:1275 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 5 47 theme plasma 877:882 arg1 membrane 884:891 the plasma membrane 873:891 the plasma membrane 873:891 However, they are still transported to the plasma membrane. 17065148 7 48 gly glycosylation 1145:1157 arg2 site 1159:1162 the relevant N-linked glycosylation site 1123:1162 the relevant N-linked glycosylation site 1123:1162 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 6 49 theme C-terminal 980:989 arg1 region 991:996 the membrane-attached C-terminal region 958:996 the membrane-attached C-terminal region of TRPM8 958:1005 Furthermore, wild-type currents can be suppressed by expressing the membrane-attached C-terminal region of TRPM8. 17065148 3 50 theme TRPM8 485:489 arg1 channels 491:498 TRPM8 channels 485:498 TRPM8 channels 485:498 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 1 51 theme overall 222:228 arg1 size 230:233 their overall size 216:233 their overall size (>1000 amino acids) 216:253 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 1 51 theme overall 222:228 arg1 acids 248:252 >1000 amino acids 236:252 >1000 amino acids 236:252 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 3 52 theme intracellular 571:583 arg1 terminus 587:594 the intracellular C terminus 567:594 the intracellular C terminus 567:594 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 1 53 theme many 141:144 arg1 members 152:158 many other members 141:158 many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 141:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 7 54 theme efficient 1310:1318 arg1 multimerization 1320:1334 efficient multimerization 1310:1334 efficient multimerization 1310:1334 To separate assembly from trafficking, we investigated the maturation of TRPM8 protein by identifying and mutating the relevant N-linked glycosylation site and showing that glycosylation is neither essential for multimerization nor for transport to the plasma membrane per se but appears to facilitate efficient multimerization and transport. 17065148 1 55 theme size 230:233 arg1 predictions 320:330 their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions 216:330 their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 216:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 17065148 2 56 theme C-terminal 430:439 arg1 region 453:458 a prominent C-terminal coiled coil region 418:458 a prominent C-terminal coiled coil region 418:458 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 2 56 theme C-terminal 430:439 arg1 member 408:413 Common to each TRPM member 388:413 Common to each TRPM member 388:413 Common to each TRPM member is a prominent C-terminal coiled coil region. 17065148 3 57 theme C 585:585 arg1 terminus 587:594 the intracellular C terminus 567:594 the intracellular C terminus 567:594 Here we have shown that TRPM8 channels assemble as multimers using the putative coiled coil region within the intracellular C terminus and that this assembly can be disturbed by a single point mutation within the coiled coil region. 17065148 1 58 theme other 146:150 arg1 members 152:158 many other members 141:158 many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions 141:385 TRPM (transient receptor potential melastatin-like) channels are distinct from many other members of the transient receptor potential family in regard to their overall size (>1000 amino acids), the lack of N-terminal ankyrin-like repeats, and hydrophobicity predictions that may allow for more than six transmembrane regions. 3484703 6 0 theme % 829:829 arg1 identity 831:838 74% identity 827:838 74% identity 827:838 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 6 0 theme % 829:829 arg1 kininogen 787:795 human high-Mr kininogen 773:795 human high-Mr kininogen 773:795 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 5 1 theme additional 616:625 arg1 site 651:654 one additional potential glycosylation site 612:654 one additional potential glycosylation site devoid of a sugar side chain 612:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 6 2 theme homology 736:743 arg1 degree 726:731 a high degree 719:731 a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%) 719:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 9 3 theme kininogens 1394:1403 arg1 chain 1379:1383 the heavy chain 1369:1383 the heavy chain of human kininogens 1369:1403 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 4 4 theme total 386:390 arg1 molecule 402:409 the total kininogen molecule 382:409 the total kininogen molecule 382:409 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 4 4 theme total 386:390 arg1 362 377:379 362 377:379 362 377:379 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 5 5 theme heavy 485:489 arg1 chain 491:495 the heavy chain 481:495 the heavy chain 481:495 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 1 6 theme gene 160:163 arg1 triplication 165:176 gene triplication 160:176 gene triplication 160:176 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 6 7 theme high-Mr 808:814 arg1 kininogen 816:824 bovine high-Mr kininogen 801:824 bovine high-Mr kininogen 801:824 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 4 8 theme heavy 356:360 arg1 chain 362:366 The heavy chain 352:366 The heavy chain 352:366 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 5 9 dep linked 527:532 arg1 N-glycosidically 510:525 N-glycosidically 510:525 N-glycosidically 510:525 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 8 10 theme reactive 1134:1141 arg1 sites 1143:1147 the reactive sites 1130:1147 the reactive sites for cysteine proteinase inhibition 1130:1182 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 11 theme side 455:458 arg1 chains 460:465 Three carbohydrate side chains 436:465 Three carbohydrate side chains 436:465 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 8 12 theme chain 1095:1099 arg1 segments 1101:1108 Two heavy chain segments 1085:1108 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition 1085:1182 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 1 13 theme heavy 114:118 arg1 chain 120:124 the entire heavy chain 103:124 the entire heavy chain 103:124 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 8 14 theme proteinase 1162:1171 arg1 inhibition 1173:1182 cysteine proteinase inhibition 1153:1182 cysteine proteinase inhibition 1153:1182 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 7 15 theme low-Mr 950:955 arg1 kininogen 957:965 human low-Mr kininogen 944:965 human low-Mr kininogen predicted from its cDNA sequence 944:998 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 2 16 theme heavy 210:214 arg1 chain 216:220 the heavy chain 206:220 the heavy chain of human high-molecular-mass kininogen 206:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 7 17 theme human 907:911 arg1 kininogen 921:929 human high-Mr kininogen 907:929 human high-Mr kininogen 907:929 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 9 18 theme human 1388:1392 arg1 kininogens 1394:1403 human kininogens 1388:1403 human kininogens 1388:1403 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 6 19 theme T-kininogen 849:859 arg1 chains 763:768 the heavy chains 753:768 the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%) 753:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 5 20 located found 472:476 arg2 chains 460:465 Three carbohydrate side chains 436:465 Three carbohydrate side chains 436:465 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 20 located found 472:476 arg1 chain 491:495 the heavy chain 481:495 the heavy chain 481:495 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 21 theme glycosylation 637:649 arg1 site 651:654 one additional potential glycosylation site 612:654 one additional potential glycosylation site devoid of a sugar side chain 612:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 7 22 theme structure 894:902 arg1 Comparison 868:877 Comparison 868:877 Comparison 868:877 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 5 23 from present 558:564 arg1 Asn-Xaa-Thr 589:599 the acceptor sequon Asn-Xaa-Thr 569:599 the acceptor sequon Asn-Xaa-Thr (or -Ser) 569:609 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 23 from present 558:564 arg1 -Ser 605:608 -Ser 605:608 -Ser 605:608 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 8 24 theme extensive 1192:1200 arg1 homology 1211:1218 an extensive sequence homology 1189:1218 an extensive sequence homology with other mammalian cysteine proteinase inhibitors 1189:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 25 theme devoid 656:661 arg1 site 651:654 one additional potential glycosylation site 612:654 one additional potential glycosylation site devoid of a sugar side chain 612:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 2 26 theme high-molecular-mass 231:249 arg1 kininogen 251:259 human high-molecular-mass kininogen 225:259 human high-molecular-mass kininogen 225:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 4 27 theme amino 415:419 arg1 residues 426:433 362, the total kininogen molecule 626 amino acid residues 377:433 362, the total kininogen molecule 626 amino acid residues 377:433 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 5 28 gly glycosylation 637:649 arg2 one 612:614 one 612:614 one 612:614 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 28 gly glycosylation 637:649 arg2 position 697:704 position 30 697:707 position 30 697:707 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 28 gly glycosylation 637:649 arg2 site 651:654 one additional potential glycosylation site 612:654 one additional potential glycosylation site devoid of a sugar side chain 612:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 0 29 theme kininogen 65:73 arg1 structure 26:34 the primary structure 14:34 the primary structure of human high-molecular-mass kininogen 14:73 Completion of the primary structure of human high-molecular-mass kininogen. 3484703 8 30 theme other 1225:1229 arg1 inhibitors 1261:1270 other mammalian cysteine proteinase inhibitors 1225:1270 other mammalian cysteine proteinase inhibitors 1225:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 9 31 theme ancestral 1435:1443 arg1 units 1445:1449 at least two ancestral units 1422:1449 at least two ancestral units 1422:1449 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 8 32 theme cysteine 1241:1248 arg1 inhibitors 1261:1270 other mammalian cysteine proteinase inhibitors 1225:1270 other mammalian cysteine proteinase inhibitors 1225:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 33 theme acceptor 573:580 arg1 Asn-Xaa-Thr 589:599 the acceptor sequon Asn-Xaa-Thr 569:599 the acceptor sequon Asn-Xaa-Thr (or -Ser) 569:609 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 33 theme acceptor 573:580 arg1 -Ser 605:608 -Ser 605:608 -Ser 605:608 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 3 34 theme kininogen 332:340 arg1 molecule 342:349 the high-Mr kininogen molecule 320:349 the high-Mr kininogen molecule 320:349 It completes the primary structure of the high-Mr kininogen molecule. 3484703 1 35 theme amino 80:84 arg1 sequence 91:98 The amino acid sequence 76:98 The amino acid sequence of the entire heavy chain 76:124 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 4 36 contain contains 368:375 arg1 chain 362:366 The heavy chain 352:366 The heavy chain 352:366 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 4 36 contain contains 368:375 arg2 residues 426:433 362, the total kininogen molecule 626 amino acid residues 377:433 362, the total kininogen molecule 626 amino acid residues 377:433 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 9 37 theme gene 1466:1469 arg1 duplication 1471:1481 gene duplication 1466:1481 gene duplication 1466:1481 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 7 38 theme human 1042:1046 arg1 kininogens 1048:1057 the two human kininogens 1034:1057 the two human kininogens 1034:1057 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 5 39 from Asn-Xaa-Thr 589:599 arg1 present 558:564 present 558:564 present 558:564 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 6 40 theme bovine 801:806 arg1 kininogen 816:824 bovine high-Mr kininogen 801:824 bovine high-Mr kininogen 801:824 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 0 41 theme structure 26:34 arg1 Completion 0:9 Completion 0:9 Completion of the primary structure of human high-molecular-mass kininogen 0:73 Completion of the primary structure of human high-molecular-mass kininogen. 3484703 6 42 theme kininogen 787:795 arg1 chains 763:768 the heavy chains 753:768 the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%) 753:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 9 43 theme events 1494:1499 arg1 series 1456:1461 a series 1454:1461 a series of gene duplication and fusion events 1454:1499 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 0 44 theme human 39:43 arg1 kininogen 65:73 human high-molecular-mass kininogen 39:73 human high-molecular-mass kininogen 39:73 Completion of the primary structure of human high-molecular-mass kininogen. 3484703 6 45 theme human 773:777 arg1 kininogen 787:795 human high-Mr kininogen 773:795 human high-Mr kininogen 773:795 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 6 45 theme human 773:777 arg1 identity 831:838 74% identity 827:838 74% identity 827:838 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 2 46 theme acid 189:192 arg1 sequence 194:201 The amino acid sequence 179:201 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen 179:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 5 47 attach linked 527:532 arg2 them 505:508 them 505:508 them 505:508 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 47 attach linked 527:532 arg2 all 498:500 all 498:500 all 498:500 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 47 attach linked 527:532 arg1 asparagine 537:546 asparagine 537:546 asparagine 537:546 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 48 theme carbohydrate 442:453 arg1 chains 460:465 Three carbohydrate side chains 436:465 Three carbohydrate side chains 436:465 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 9 49 theme human 1299:1303 arg1 kininogen 1313:1321 human high-Mr kininogen 1299:1321 human high-Mr kininogen 1299:1321 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 0 50 theme high-molecular-mass 45:63 arg1 kininogen 65:73 human high-molecular-mass kininogen 39:73 human high-molecular-mass kininogen 39:73 Completion of the primary structure of human high-molecular-mass kininogen. 3484703 5 51 located present 558:564 arg2 asparagine 537:546 asparagine 537:546 asparagine 537:546 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 51 located present 558:564 arg1 Asn-Xaa-Thr 589:599 the acceptor sequon Asn-Xaa-Thr 569:599 the acceptor sequon Asn-Xaa-Thr (or -Ser) 569:609 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 51 located present 558:564 arg1 -Ser 605:608 -Ser 605:608 -Ser 605:608 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 9 52 theme kininogen 1313:1321 arg1 chain 1290:1294 the heavy chain 1280:1294 the heavy chain of human high-Mr kininogen 1280:1321 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 9 53 theme repetitive 1327:1336 arg1 units 1338:1342 repetitive units 1327:1342 repetitive units 1327:1342 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 4 54 theme kininogen 392:400 arg1 molecule 402:409 the total kininogen molecule 382:409 the total kininogen molecule 382:409 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 4 54 theme kininogen 392:400 arg1 362 377:379 362 377:379 362 377:379 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 7 55 theme cDNA 986:989 arg1 sequence 991:998 its cDNA sequence 982:998 its cDNA sequence 982:998 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 6 56 theme kininogen 816:824 arg1 chains 763:768 the heavy chains 753:768 the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%) 753:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 6 57 theme high 721:724 arg1 degree 726:731 a high degree 719:731 a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%) 719:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 4 58 theme 362 377:379 arg1 residues 426:433 362, the total kininogen molecule 626 amino acid residues 377:433 362, the total kininogen molecule 626 amino acid residues 377:433 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 9 59 theme heavy 1373:1377 arg1 chain 1379:1383 the heavy chain 1369:1383 the heavy chain of human kininogens 1369:1403 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 7 60 theme human 944:948 arg1 kininogen 957:965 human low-Mr kininogen 944:965 human low-Mr kininogen predicted from its cDNA sequence 944:998 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 2 61 theme chain 216:220 arg1 sequence 194:201 The amino acid sequence 179:201 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen 179:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 8 62 theme heavy 1089:1093 arg1 segments 1101:1108 Two heavy chain segments 1085:1108 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition 1085:1182 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 8 63 theme cysteine 1153:1160 arg1 inhibition 1173:1182 cysteine proteinase inhibition 1153:1182 cysteine proteinase inhibition 1153:1182 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 64 attach present 558:564 arg2 asparagine 537:546 asparagine 537:546 asparagine 537:546 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 64 attach present 558:564 arg1 Asn-Xaa-Thr 589:599 the acceptor sequon Asn-Xaa-Thr 569:599 the acceptor sequon Asn-Xaa-Thr (or -Ser) 569:609 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 64 attach present 558:564 arg1 -Ser 605:608 -Ser 605:608 -Ser 605:608 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 65 theme potential 627:635 arg1 site 651:654 one additional potential glycosylation site 612:654 one additional potential glycosylation site devoid of a sugar side chain 612:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 7 66 theme high-Mr 913:919 arg1 kininogen 921:929 human high-Mr kininogen 907:929 human high-Mr kininogen 907:929 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 1 67 theme entire 107:112 arg1 chain 120:124 the entire heavy chain 103:124 the entire heavy chain 103:124 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 6 68 theme rat 845:847 arg1 T-kininogen 849:859 rat T-kininogen 845:859 rat T-kininogen (61%) 845:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 6 68 theme rat 845:847 arg1 % 864:864 61% 862:864 61% 862:864 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 3 69 theme primary 299:305 arg1 structure 307:315 the primary structure 295:315 the primary structure of the high-Mr kininogen molecule 295:349 It completes the primary structure of the high-Mr kininogen molecule. 3484703 7 70 theme kininogen 921:929 arg1 structure 894:902 the primary structure 882:902 the primary structure of human high-Mr kininogen 882:929 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 2 71 theme kininogen 251:259 arg1 chain 216:220 the heavy chain 206:220 the heavy chain of human high-molecular-mass kininogen 206:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 1 72 theme chain 120:124 arg1 sequence 91:98 The amino acid sequence 76:98 The amino acid sequence of the entire heavy chain 76:124 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 1 72 theme chain 120:124 arg1 evidence 130:137 evidence 130:137 evidence for its evolution by gene triplication 130:176 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 7 73 with Comparison 868:877 arg1 that 936:939 that 936:939 that 936:939 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 4 74 theme acid 421:424 arg1 residues 426:433 362, the total kininogen molecule 626 amino acid residues 377:433 362, the total kininogen molecule 626 amino acid residues 377:433 The heavy chain contains 362, the total kininogen molecule 626 amino acid residues. 3484703 2 75 theme human 225:229 arg1 kininogen 251:259 human high-molecular-mass kininogen 225:259 human high-molecular-mass kininogen 225:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 7 76 theme primary 886:892 arg1 structure 894:902 the primary structure 882:902 the primary structure of human high-Mr kininogen 882:929 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 3 77 theme high-Mr 324:330 arg1 molecule 342:349 the high-Mr kininogen molecule 320:349 the high-Mr kininogen molecule 320:349 It completes the primary structure of the high-Mr kininogen molecule. 3484703 8 78 theme sequence 1202:1209 arg1 homology 1211:1218 an extensive sequence homology 1189:1218 an extensive sequence homology with other mammalian cysteine proteinase inhibitors 1189:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 79 located found 688:692 arg1 position 697:704 position 30 697:707 position 30 697:707 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 79 located found 688:692 arg2 site 651:654 one additional potential glycosylation site 612:654 one additional potential glycosylation site devoid of a sugar side chain 612:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 8 80 with homology 1211:1218 arg1 inhibitors 1261:1270 other mammalian cysteine proteinase inhibitors 1225:1270 other mammalian cysteine proteinase inhibitors 1225:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 81 theme chain 679:683 arg1 devoid 656:661 devoid 656:661 devoid 656:661 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 8 82 theme mammalian 1231:1239 arg1 inhibitors 1261:1270 other mammalian cysteine proteinase inhibitors 1225:1270 other mammalian cysteine proteinase inhibitors 1225:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 5 83 theme sequon 582:587 arg1 Asn-Xaa-Thr 589:599 the acceptor sequon Asn-Xaa-Thr 569:599 the acceptor sequon Asn-Xaa-Thr (or -Ser) 569:609 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 5 83 theme sequon 582:587 arg1 -Ser 605:608 -Ser 605:608 -Ser 605:608 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 3 84 theme molecule 342:349 arg1 structure 307:315 the primary structure 295:315 the primary structure of the high-Mr kininogen molecule 295:349 It completes the primary structure of the high-Mr kininogen molecule. 3484703 8 85 theme proteinase 1250:1259 arg1 inhibitors 1261:1270 other mammalian cysteine proteinase inhibitors 1225:1270 other mammalian cysteine proteinase inhibitors 1225:1270 Two heavy chain segments believed to contain the reactive sites for cysteine proteinase inhibition show an extensive sequence homology with other mammalian cysteine proteinase inhibitors. 3484703 7 86 theme kininogens 1048:1057 arg1 chains 1024:1029 the heavy chains 1014:1029 the heavy chains of the two human kininogens 1014:1057 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 7 86 theme kininogens 1048:1057 arg1 identical 1074:1082 identical 1074:1082 identical 1074:1082 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 1 87 theme acid 86:89 arg1 sequence 91:98 The amino acid sequence 76:98 The amino acid sequence of the entire heavy chain 76:124 The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication. 3484703 5 88 theme sugar 668:672 arg1 chain 679:683 a sugar side chain 666:683 a sugar side chain 666:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 0 89 theme primary 18:24 arg1 structure 26:34 the primary structure 14:34 the primary structure of human high-molecular-mass kininogen 14:73 Completion of the primary structure of human high-molecular-mass kininogen. 3484703 9 90 theme duplication 1471:1481 arg1 series 1456:1461 a series 1454:1461 a series of gene duplication and fusion events 1454:1499 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 9 91 theme fusion 1487:1492 arg1 events 1494:1499 fusion events 1487:1499 fusion events 1487:1499 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 7 92 theme heavy 1018:1022 arg1 chains 1024:1029 the heavy chains 1014:1029 the heavy chains of the two human kininogens 1014:1057 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 7 92 theme heavy 1018:1022 arg1 identical 1074:1082 identical 1074:1082 identical 1074:1082 Comparison of the primary structure of human high-Mr kininogen with that of human low-Mr kininogen predicted from its cDNA sequence, reveals that the heavy chains of the two human kininogens are completely identical. 3484703 6 93 theme high-Mr 779:785 arg1 kininogen 787:795 human high-Mr kininogen 773:795 human high-Mr kininogen 773:795 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 6 93 theme high-Mr 779:785 arg1 identity 831:838 74% identity 827:838 74% identity 827:838 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 5 94 theme side 674:677 arg1 chain 679:683 a sugar side chain 666:683 a sugar side chain 666:683 Three carbohydrate side chains were found in the heavy chain, all of them N-glycosidically linked to asparagine, which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser); one additional potential glycosylation site devoid of a sugar side chain is found at position 30. 3484703 9 95 theme heavy 1284:1288 arg1 chain 1290:1294 the heavy chain 1280:1294 the heavy chain of human high-Mr kininogen 1280:1321 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 3484703 2 96 theme amino 183:187 arg1 sequence 194:201 The amino acid sequence 179:201 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen 179:259 The amino acid sequence of the heavy chain of human high-molecular-mass kininogen has been determined. 3484703 6 97 theme heavy 757:761 arg1 chains 763:768 the heavy chains 753:768 the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%) 753:865 There is a high degree of homology between the heavy chains of human high-Mr kininogen and bovine high-Mr kininogen (74% identity), or rat T-kininogen (61%). 3484703 9 98 theme high-Mr 1305:1311 arg1 kininogen 1313:1321 human high-Mr kininogen 1299:1321 human high-Mr kininogen 1299:1321 Within the heavy chain of human high-Mr kininogen are repetitive units strongly suggesting that the heavy chain of human kininogens has evolved from at least two ancestral units by a series of gene duplication and fusion events. 8757293 9 0 theme granulomatosis 1514:1527 arg1 autoantibodies 1529:1542 Wegener's granulomatosis autoantibodies 1504:1542 Wegener's granulomatosis autoantibodies 1504:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 7 1 theme protein 1271:1277 arg1 interior 1255:1262 the interior 1251:1262 the interior of the protein 1251:1277 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 1 2 theme molecular 248:256 arg1 replacement 258:268 molecular replacement 248:268 molecular replacement using the human leukocyte elastase structure 248:313 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 7 3 from position 1138:1145 arg1 substitution 1108:1119 The substitution 1104:1119 The substitution of Ala by Asp at position 213 at the back of S1 1104:1167 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 9 4 theme structure 1586:1594 arg1 regions 1553:1559 regions 1553:1559 regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease 1553:1684 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 2 5 theme A 485:485 arg1 resolution 487:496 10.0 to 2.2 A resolution 473:496 10.0 to 2.2 A resolution 473:496 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 7 6 theme side-chain 1223:1232 arg1 specificity 1191:1201 its specificity 1187:1201 its specificity 1187:1201 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 7 6 theme side-chain 1223:1232 arg1 points 1234:1239 the Asp side-chain points 1215:1239 the Asp side-chain points back into the interior of the protein 1215:1277 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 5 7 theme substrate 768:776 arg1 sites 786:790 the substrate binding sites 764:790 the substrate binding sites 764:790 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 7 theme substrate 768:776 arg1 polar 813:817 polar 813:817 polar 813:817 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 7 theme substrate 768:776 arg1 S4 793:794 S4 793:794 S4 to S3' 793:801 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 4 8 theme overall 631:637 arg1 fold 639:642 The overall fold 627:642 The overall fold 627:642 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 3 9 theme space 530:534 arg1 group 536:540 space group 530:540 space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da) 530:624 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 5 10 from general 755:761 arg1 sites 786:790 the substrate binding sites 764:790 the substrate binding sites 764:790 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 10 from general 755:761 arg1 polar 813:817 polar 813:817 polar 813:817 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 6 11 theme elastase 1084:1091 arg1 structure 1093:1101 the elastase structure 1080:1101 the elastase structure 1080:1101 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 0 12 theme serine 43:48 arg1 antigen 61:67 a neutrophil serine proteinase antigen 30:67 a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies 30:106 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 0 12 theme serine 43:48 arg1 PR3 25:27 PR3 25:27 PR3 25:27 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 1 13 theme PR3 134:136 arg1 structure 121:129 The crystal structure 109:129 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, 109:227 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 8 14 theme N-linked 1328:1335 arg1 2-acetamido-2-deoxy-beta-D-glucopyranose 1337:1376 N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose 1328:1376 N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose 1328:1376 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 4 15 theme structures 683:692 arg1 domains 660:666 two domains 656:666 two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases 656:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 4 15 theme structures 683:692 arg1 structures 683:692 beta-barrel structures 671:692 beta-barrel structures typical of the chymotrypsin family of serine proteinases 671:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 2 16 theme sigma 369:373 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 0 17 theme granulomatosis 82:95 arg1 antibodies 97:106 Wegener's granulomatosis antibodies 72:106 Wegener's granulomatosis antibodies 72:106 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 5 18 from polar 813:817 arg1 general 755:761 general 755:761 general 755:761 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 19 theme human 868:872 arg1 proteinase 856:865 the related proteinase 844:865 the related proteinase 844:865 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 19 theme human 868:872 arg1 elastase 884:891 human leukocyte elastase 868:891 human leukocyte elastase 868:891 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 2 20 theme Fo 431:432 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 9 21 theme linear 1451:1456 arg1 PR3 1477:1479 PR3 1477:1479 PR3 reported to react with Wegener's granulomatosis autoantibodies 1477:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 9 21 theme linear 1451:1456 arg1 sites 1468:1472 The linear antigenic sites 1447:1472 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies 1447:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 2 22 theme of-Fc 397:401 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 6 23 theme small 944:948 arg1 residues 960:967 small aliphatic residues 944:967 small aliphatic residues at the P1 position of a substrate 944:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 4 24 theme proteinases 739:749 arg1 family 722:727 the chymotrypsin family 705:727 the chymotrypsin family of serine proteinases 705:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 2 25 theme Fo 384:385 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 5 26 theme binding 778:784 arg1 sites 786:790 the substrate binding sites 764:790 the substrate binding sites 764:790 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 26 theme binding 778:784 arg1 polar 813:817 polar 813:817 polar 813:817 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 26 theme binding 778:784 arg1 S4 793:794 S4 793:794 S4 to S3' 793:801 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 1 27 theme leukocyte 286:294 arg1 elastase 296:303 human leukocyte elastase 280:303 the human leukocyte elastase structure 276:313 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 0 28 theme PR3 25:27 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 0:107 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 9 29 theme PR3 1477:1479 arg1 PR3 1477:1479 PR3 1477:1479 PR3 reported to react with Wegener's granulomatosis autoantibodies 1477:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 9 29 theme PR3 1477:1479 arg1 sites 1468:1472 The linear antigenic sites 1447:1472 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies 1447:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 6 30 theme PR3 936:938 arg1 preference 922:931 The experimentally observed preference 894:931 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate 894:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 3 31 theme asymmetric 573:582 arg1 unit 584:587 the asymmetric unit 569:587 the asymmetric unit (Vm approximately equal to 2.6 A/Da) 569:624 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 4 32 theme chymotrypsin 709:720 arg1 family 722:727 the chymotrypsin family 705:727 the chymotrypsin family of serine proteinases 705:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 4 33 theme serine 732:737 arg1 proteinases 739:749 serine proteinases 732:749 serine proteinases 732:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 2 34 theme PR3 320:322 arg1 structure 324:332 The PR3 structure 316:332 The PR3 structure 316:332 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 1 35 theme polymorphonuclear 198:214 arg1 neutrophils 216:226 human polymorphonuclear neutrophils 192:226 human polymorphonuclear neutrophils 192:226 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 8 36 link N-linked 1328:1335 arg1 2-acetamido-2-deoxy-beta-D-glucopyranose 1337:1376 N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose 1328:1376 N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose 1328:1376 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 6 37 from position 979:986 arg1 residues 960:967 small aliphatic residues 944:967 small aliphatic residues at the P1 position of a substrate 944:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 5 38 from sites 835:839 arg1 proteinase 856:865 the related proteinase 844:865 the related proteinase 844:865 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 38 from sites 835:839 arg1 elastase 884:891 human leukocyte elastase 868:891 human leukocyte elastase 868:891 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 3 39 dep unit 584:587 arg1 Vm 590:591 Vm 590:591 Vm approximately equal to 2.6 A/Da 590:623 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 3 40 with group 536:540 arg1 molecules 556:564 four molecules 551:564 four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da) 551:624 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 7 41 theme Ala 1124:1126 arg1 substitution 1108:1119 The substitution 1104:1119 The substitution of Ala by Asp at position 213 at the back of S1 1104:1167 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 2 42 from R-factor 357:364 arg1 range 464:468 the range 460:468 the range of 10.0 to 2.2 A resolution 460:496 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 1 43 theme serine 141:146 arg1 PR3 134:136 PR3 134:136 PR3 134:136 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 1 43 theme serine 141:146 arg1 proteinase 148:157 a serine proteinase 139:157 a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils 139:226 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 2 44 dep resolution 487:496 arg1 to 478:479 to 478:479 to 478:479 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 3 45 dep group 536:540 arg1 P21 542:544 P21 542:544 space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da) 530:624 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 9 46 theme three-dimensional 1568:1584 arg1 structure 1586:1594 the three-dimensional structure 1564:1594 the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease 1564:1684 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 6 47 theme Ile 1030:1032 arg1 substitution 1034:1045 Ile substitution 1030:1045 Ile substitution 1030:1045 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 1 48 theme azurophilic 168:178 arg1 granules 180:187 the azurophilic granules 164:187 the azurophilic granules of human polymorphonuclear neutrophils 164:226 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 2 49 theme resolution 487:496 arg1 range 464:468 the range 460:468 the range of 10.0 to 2.2 A resolution 460:496 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 8 50 theme PR3 1284:1286 arg1 structure 1288:1296 The PR3 structure 1280:1296 The PR3 structure 1280:1296 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 8 51 attach attached 1426:1433 arg2 unit 1322:1325 a disaccharide unit 1307:1325 a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159 1307:1444 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 8 51 attach attached 1426:1433 arg1 Asn 1438:1440 Asn 159 1438:1444 Asn 159 1438:1444 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 9 52 theme inactive 1619:1626 arg1 pro-form 1628:1635 the inactive pro-form 1615:1635 the inactive pro-form of the enzyme 1615:1649 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 8 53 theme disaccharide 1309:1320 arg1 unit 1322:1325 a disaccharide unit 1307:1325 a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159 1307:1444 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 7 54 theme Asp 1219:1221 arg1 specificity 1191:1201 its specificity 1187:1201 its specificity 1187:1201 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 7 54 theme Asp 1219:1221 arg1 points 1234:1239 the Asp side-chain points 1215:1239 the Asp side-chain points back into the interior of the protein 1215:1277 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 1 55 from granules 180:187 arg1 PR3 134:136 PR3 134:136 PR3 134:136 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 1 55 from granules 180:187 arg1 proteinase 148:157 a serine proteinase 139:157 a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils 139:226 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 1 56 theme crystal 113:119 arg1 structure 121:129 The crystal structure 109:129 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, 109:227 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 0 57 theme proteinase 50:59 arg1 antigen 61:67 a neutrophil serine proteinase antigen 30:67 a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies 30:106 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 0 57 theme proteinase 50:59 arg1 PR3 25:27 PR3 25:27 PR3 25:27 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 6 58 theme observed 913:920 arg1 preference 922:931 The experimentally observed preference 894:931 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate 894:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 9 59 theme enzyme 1644:1649 arg1 pro-form 1628:1635 the inactive pro-form 1615:1635 the inactive pro-form of the enzyme 1615:1649 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 8 60 theme 1,6-linked 1382:1391 arg1 alpha-L-fucopyranose 1393:1412 1,6-linked alpha-L-fucopyranose 1382:1412 1,6-linked alpha-L-fucopyranose 1382:1412 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 5 61 theme related 848:854 arg1 proteinase 856:865 the related proteinase 844:865 the related proteinase 844:865 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 61 theme related 848:854 arg1 elastase 884:891 human leukocyte elastase 868:891 human leukocyte elastase 868:891 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 4 62 theme beta-barrel 671:681 arg1 structures 683:692 beta-barrel structures 671:692 beta-barrel structures typical of the chymotrypsin family of serine proteinases 671:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 6 63 from position 1050:1057 arg1 Val 1023:1025 the Val 1019:1025 the Val to Ile substitution at position 190 1019:1061 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 0 64 theme antibodies 97:106 arg1 antigen 61:67 a neutrophil serine proteinase antigen 30:67 a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies 30:106 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 0 64 theme antibodies 97:106 arg1 PR3 25:27 PR3 25:27 PR3 25:27 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 6 65 theme P1 976:977 arg1 position 979:986 the P1 position 972:986 the P1 position of a substrate 972:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 9 66 theme disease 1678:1684 arg1 pathogenesis 1658:1669 the pathogenesis 1654:1669 the pathogenesis of the disease 1654:1684 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 4 67 theme typical 694:700 arg1 structures 683:692 beta-barrel structures 671:692 beta-barrel structures typical of the chymotrypsin family of serine proteinases 671:749 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 8757293 3 68 from molecules 556:564 arg1 unit 584:587 the asymmetric unit 569:587 the asymmetric unit (Vm approximately equal to 2.6 A/Da) 569:624 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 2 69 theme = 367:367 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 0 70 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 0:107 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 5 71 theme comparable 824:833 arg1 sites 835:839 comparable sites 824:839 comparable sites in the related proteinase, human leukocyte elastase 824:891 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 8 72 dep unit 1322:1325 arg1 2-acetamido-2-deoxy-beta-D-glucopyranose 1337:1376 N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose 1328:1376 N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose 1328:1376 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 8 72 dep unit 1322:1325 arg1 alpha-L-fucopyranose 1393:1412 1,6-linked alpha-L-fucopyranose 1382:1412 1,6-linked alpha-L-fucopyranose 1382:1412 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 9 73 theme antigenic 1458:1466 arg1 PR3 1477:1479 PR3 1477:1479 PR3 reported to react with Wegener's granulomatosis autoantibodies 1477:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 9 73 theme antigenic 1458:1466 arg1 sites 1468:1472 The linear antigenic sites 1447:1472 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies 1447:1542 The linear antigenic sites of PR3 reported to react with Wegener's granulomatosis autoantibodies occur in regions of the three-dimensional structure that may implicate the inactive pro-form of the enzyme in the pathogenesis of the disease. 8757293 6 74 theme aliphatic 950:958 arg1 residues 960:967 small aliphatic residues 944:967 small aliphatic residues at the P1 position of a substrate 944:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 2 75 theme parallel/sigma 403:416 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 2 76 theme magnitude 387:395 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 1 77 theme human 280:284 arg1 elastase 296:303 human leukocyte elastase 280:303 the human leukocyte elastase structure 276:313 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 2 78 theme 0.201 438:442 arg1 R-factor 357:364 an R-factor 354:364 an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution 354:496 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 5 79 dep S4 793:794 arg1 to 796:797 to 796:797 to 796:797 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 2 80 theme parallel 375:382 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 0 81 theme neutrophil 32:41 arg1 antigen 61:67 a neutrophil serine proteinase antigen 30:67 a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies 30:106 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 0 81 theme neutrophil 32:41 arg1 PR3 25:27 PR3 25:27 PR3 25:27 The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies. 8757293 1 82 theme elastase 296:303 arg1 structure 305:313 the human leukocyte elastase structure 276:313 the human leukocyte elastase structure 276:313 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 7 83 from back 1158:1161 arg1 substitution 1108:1119 The substitution 1104:1119 The substitution of Ala by Asp at position 213 at the back of S1 1104:1167 The substitution of Ala by Asp at position 213 at the back of S1 should not affect its specificity greatly, as the Asp side-chain points back into the interior of the protein. 8757293 8 84 link 1,6-linked 1382:1391 arg1 alpha-L-fucopyranose 1393:1412 1,6-linked alpha-L-fucopyranose 1382:1412 1,6-linked alpha-L-fucopyranose 1382:1412 The PR3 structure includes a disaccharide unit (N-linked 2-acetamido-2-deoxy-beta-D-glucopyranose and 1,6-linked alpha-L-fucopyranose) covalently attached to Asn 159. 8757293 3 85 theme equal 607:611 arg1 Vm 590:591 Vm 590:591 Vm approximately equal to 2.6 A/Da 590:623 The enzyme was crystallized in space group P21 with four molecules in the asymmetric unit (Vm approximately equal to 2.6 A/Da). 8757293 5 86 theme leukocyte 874:882 arg1 proteinase 856:865 the related proteinase 844:865 the related proteinase 844:865 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 5 86 theme leukocyte 874:882 arg1 elastase 884:891 human leukocyte elastase 868:891 human leukocyte elastase 868:891 In general, the substrate binding sites, S4 to S3', are more polar than comparable sites in the related proteinase, human leukocyte elastase. 8757293 2 87 dep R-factor 357:364 arg1 magnitude 418:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude 367:426 = sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo 367:432 The PR3 structure has been refined to an R-factor (= sigma parallel Fo magnitude of-Fc parallel/sigma magnitude of Fo) of 0.201 for all data in the range of 10.0 to 2.2 A resolution. 8757293 1 88 theme neutrophils 216:226 arg1 granules 180:187 the azurophilic granules 164:187 the azurophilic granules of human polymorphonuclear neutrophils 164:226 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 1 89 theme human 192:196 arg1 neutrophils 216:226 human polymorphonuclear neutrophils 192:226 human polymorphonuclear neutrophils 192:226 The crystal structure of PR3, a serine proteinase from the azurophilic granules of human polymorphonuclear neutrophils, has been solved by molecular replacement using the human leukocyte elastase structure. 8757293 6 90 theme substrate 993:1001 arg1 position 979:986 the P1 position 972:986 the P1 position of a substrate 972:1001 The experimentally observed preference of PR3 for small aliphatic residues at the P1 position of a substrate is explained by the Val to Ile substitution at position 190 when compared to the elastase structure. 8757293 4 91 theme family 722:727 arg1 typical 694:700 typical 694:700 typical 694:700 The overall fold consists of two domains of beta-barrel structures typical of the chymotrypsin family of serine proteinases. 2071582 9 0 theme stringency 1495:1504 arg1 hybridization 1506:1518 High stringency hybridization 1490:1518 High stringency hybridization of chymase DNA to a human genomic DNA blot 1490:1561 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 6 1 theme cathespin 1113:1121 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 2 from placement 901:909 arg1 organization 927:938 the organization 923:938 the organization of this human chymase gene 923:965 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 2 from placement 901:909 arg1 similar 970:976 similar 970:976 similar 970:976 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 2 3 theme catalytic 302:310 arg1 domain 312:317 a 226-amino acid catalytic domain 285:317 a 226-amino acid catalytic domain 285:317 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 7 4 theme substantial 1300:1310 arg1 distance 1325:1332 substantial evolutionary distance 1300:1332 substantial evolutionary distance 1300:1332 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 6 5 theme rat 1060:1062 arg1 chymase 1064:1070 rat chymase II 1060:1073 rat chymase II 1060:1073 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 7 6 theme mast 1256:1259 arg1 proteases 1273:1281 the major mast cell serine proteases 1246:1281 the major mast cell serine proteases 1246:1281 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 4 7 theme amino 510:514 arg1 sequence 521:528 NH2-terminal amino acid sequence 497:528 NH2-terminal amino acid sequence reported for human skin chymase 497:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 9 8 theme DNA 1531:1533 arg1 hybridization 1506:1518 High stringency hybridization 1490:1518 High stringency hybridization of chymase DNA to a human genomic DNA blot 1490:1561 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 6 9 theme several 989:995 arg1 proteases 1039:1047 several other granule-associated leukocyte serine proteases 989:1047 several other granule-associated leukocyte serine proteases 989:1047 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 9 theme several 989:995 arg1 granzymes 1087:1095 lymphocyte granzymes 1076:1095 lymphocyte granzymes 1076:1095 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 9 theme several 989:995 arg1 chymase 1064:1070 rat chymase II 1060:1073 rat chymase II 1060:1073 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 9 theme several 989:995 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 5 10 theme sequence 759:766 arg1 identity 768:775 sequence identity 759:775 sequence identity to dog mast cell chymase (83%) 759:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 10 11 from skin 1749:1752 arg1 amplification 1706:1718 the amplification 1702:1718 the amplification of chymase-specific DNA from skin and placental cDNA libraries 1702:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 10 11 from skin 1749:1752 arg1 DNA 1740:1742 chymase-specific DNA 1723:1742 chymase-specific DNA from skin and placental cDNA libraries 1723:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 6 12 from phase 891:895 arg1 organization 927:938 the organization 923:938 the organization of this human chymase gene 923:965 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 12 from phase 891:895 arg1 similar 970:976 similar 970:976 similar 970:976 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 9 13 theme genomic 1546:1552 arg1 blot 1558:1561 a human genomic DNA blot 1538:1561 a human genomic DNA blot 1538:1561 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 4 14 theme skin 549:552 arg1 chymase 554:560 human skin chymase 543:560 human skin chymase 543:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 0 15 theme cell 92:95 arg1 chymase 97:103 mast cell chymase 87:103 mast cell chymase 87:103 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 2 16 theme 2-amino 256:262 arg1 propeptide 269:278 an acidic 2-amino acid propeptide 246:278 an acidic 2-amino acid propeptide 246:278 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 10 17 from libraries 1773:1781 arg1 amplification 1706:1718 the amplification 1702:1718 the amplification of chymase-specific DNA from skin and placental cDNA libraries 1702:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 10 17 from libraries 1773:1781 arg1 DNA 1740:1742 chymase-specific DNA 1723:1742 chymase-specific DNA from skin and placental cDNA libraries 1723:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 2 18 with preproenzyme 196:207 arg1 domain 312:317 a 226-amino acid catalytic domain 285:317 a 226-amino acid catalytic domain 285:317 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 2 18 with preproenzyme 196:207 arg1 peptide 237:243 a 19-amino acid signal peptide 214:243 a 19-amino acid signal peptide 214:243 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 2 18 with preproenzyme 196:207 arg1 propeptide 269:278 an acidic 2-amino acid propeptide 246:278 an acidic 2-amino acid propeptide 246:278 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 8 19 from Amplification 1335:1347 arg1 DNA 1420:1422 hamster/human hybrid cell line DNA 1389:1422 hamster/human hybrid cell line DNA 1389:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 7 20 theme mast 1191:1194 arg1 tryptase 1201:1208 mast cell tryptase 1191:1208 mast cell tryptase 1191:1208 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 4 21 theme amino 629:633 arg1 sequence 640:647 NH2-terminal and internal amino acid sequence 603:647 NH2-terminal and internal amino acid sequence reported for human heart chymase 603:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 6 22 theme granule-associated 1003:1020 arg1 proteases 1039:1047 several other granule-associated leukocyte serine proteases 989:1047 several other granule-associated leukocyte serine proteases 989:1047 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 22 theme granule-associated 1003:1020 arg1 granzymes 1087:1095 lymphocyte granzymes 1076:1095 lymphocyte granzymes 1076:1095 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 22 theme granule-associated 1003:1020 arg1 chymase 1064:1070 rat chymase II 1060:1073 rat chymase II 1060:1073 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 22 theme granule-associated 1003:1020 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 23 theme serine 1032:1037 arg1 proteases 1039:1047 several other granule-associated leukocyte serine proteases 989:1047 several other granule-associated leukocyte serine proteases 989:1047 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 23 theme serine 1032:1037 arg1 granzymes 1087:1095 lymphocyte granzymes 1076:1095 lymphocyte granzymes 1076:1095 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 23 theme serine 1032:1037 arg1 chymase 1064:1070 rat chymase II 1060:1073 rat chymase II 1060:1073 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 23 theme serine 1032:1037 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 0 24 theme acid 53:56 arg1 sequence 58:65 deduced amino acid sequence 39:65 deduced amino acid sequence of a human gene for mast cell chymase 39:103 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 5 25 theme domain 728:733 arg1 sequence 702:709 The full predicted sequence 683:709 The full predicted sequence of the catalytic domain 683:733 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 26 theme mast 784:787 arg1 chymase 794:800 dog mast cell chymase 780:800 dog mast cell chymase (83%) 780:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 26 theme mast 784:787 arg1 % 805:805 83% 803:805 83% 803:805 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 9 27 theme human 1605:1609 arg1 gene 1619:1622 more than one human chymase gene 1591:1622 more than one human chymase gene 1591:1622 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 9 28 theme gene 1619:1622 arg1 possibility 1576:1586 the possibility 1572:1586 the possibility of more than one human chymase gene 1572:1622 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 2 29 theme acid 225:228 arg1 peptide 237:243 a 19-amino acid signal peptide 214:243 a 19-amino acid signal peptide 214:243 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 4 30 theme heart 668:672 arg1 chymase 674:680 human heart chymase 662:680 human heart chymase 662:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 0 31 theme human 72:76 arg1 gene 78:81 a human gene 70:81 a human gene 70:81 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 6 32 theme other 997:1001 arg1 proteases 1039:1047 several other granule-associated leukocyte serine proteases 989:1047 several other granule-associated leukocyte serine proteases 989:1047 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 32 theme other 997:1001 arg1 granzymes 1087:1095 lymphocyte granzymes 1076:1095 lymphocyte granzymes 1076:1095 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 32 theme other 997:1001 arg1 chymase 1064:1070 rat chymase II 1060:1073 rat chymase II 1060:1073 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 32 theme other 997:1001 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 5 33 theme identity 829:836 arg1 level 820:824 a lower level 812:824 a lower level of identity to the sequences of rodent chymases (58-62%) 812:881 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 33 theme identity 829:836 arg1 level 750:754 a high level 743:754 a high level of sequence identity to dog mast cell chymase (83%) 743:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 10 34 dep Evidence 1625:1632 arg1 expressed 1659:1667 expressed 1659:1667 is expressed in human tissues 1656:1684 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 8 35 theme gene-specific 1360:1372 arg1 fragments 1374:1382 chymase gene-specific fragments 1352:1382 chymase gene-specific fragments from hamster/human hybrid cell line DNA 1352:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 6 36 dep phase 891:895 arg1 the 887:889 the 887:889 the 887:889 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 3 37 dep cationic 357:364 arg1 charge 371:376 net charge 367:376 net charge of +13 367:383 The mature enzyme is predicted to be cationic (net charge of +13) and to be modified by N-glycosylation at two sites. 2071582 4 38 theme sequence 640:647 arg1 residues 591:598 29 of 31 residues 582:598 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase 582:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 4 38 theme sequence 640:647 arg1 sequence 640:647 NH2-terminal and internal amino acid sequence 603:647 NH2-terminal and internal amino acid sequence reported for human heart chymase 603:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 3 39 theme net 367:369 arg1 charge 371:376 net charge 367:376 net charge of +13 367:383 The mature enzyme is predicted to be cationic (net charge of +13) and to be modified by N-glycosylation at two sites. 2071582 3 40 theme mature 324:329 arg1 enzyme 331:336 The mature enzyme 320:336 The mature enzyme 320:336 The mature enzyme is predicted to be cationic (net charge of +13) and to be modified by N-glycosylation at two sites. 2071582 8 41 theme hybrid 1403:1408 arg1 line 1415:1418 hamster/human hybrid cell line 1389:1418 hamster/human hybrid cell line DNA 1389:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 0 42 theme chromosomal 11:21 arg1 assignment 23:32 chromosomal assignment 11:32 chromosomal assignment 11:32 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 8 43 theme line 1415:1418 arg1 DNA 1420:1422 hamster/human hybrid cell line DNA 1389:1422 hamster/human hybrid cell line DNA 1389:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 5 44 theme full 687:690 arg1 sequence 702:709 The full predicted sequence 683:709 The full predicted sequence of the catalytic domain 683:733 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 4 45 theme amino 442:446 arg1 sequence 453:460 The amino acid sequence 438:460 The amino acid sequence 438:460 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 4 45 theme amino 442:446 arg1 identical 569:577 identical 569:577 identical 569:577 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 4 45 theme amino 442:446 arg1 identical 465:473 identical 465:473 identical 465:473 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 2 46 theme acid 297:300 arg1 domain 312:317 a 226-amino acid catalytic domain 285:317 a 226-amino acid catalytic domain 285:317 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 0 47 theme deduced 39:45 arg1 sequence 58:65 deduced amino acid sequence 39:65 deduced amino acid sequence of a human gene for mast cell chymase 39:103 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 6 48 theme chymase 954:960 arg1 gene 962:965 this human chymase gene 943:965 this human chymase gene 943:965 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 2 49 theme protein-coding 166:179 arg1 exons 181:185 The protein-coding exons 162:185 The protein-coding exons 162:185 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 8 50 theme chymase 1453:1459 arg1 gene 1461:1464 the chymase gene 1449:1464 the chymase gene 1449:1464 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 0 51 theme amino 47:51 arg1 sequence 58:65 deduced amino acid sequence 39:65 deduced amino acid sequence of a human gene for mast cell chymase 39:103 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 7 52 theme evolutionary 1312:1323 arg1 distance 1325:1332 substantial evolutionary distance 1300:1332 substantial evolutionary distance 1300:1332 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 5 53 theme chymases 865:872 arg1 sequences 845:853 the sequences 841:853 the sequences of rodent chymases (58-62%) 841:881 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 10 54 theme human 1672:1676 arg1 tissues 1678:1684 human tissues 1672:1684 human tissues 1672:1684 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 8 55 from DNA 1420:1422 arg1 fragments 1374:1382 chymase gene-specific fragments 1352:1382 chymase gene-specific fragments from hamster/human hybrid cell line DNA 1352:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 8 55 from DNA 1420:1422 arg1 Amplification 1335:1347 Amplification 1335:1347 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA 1335:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 6 56 theme neutrophil 1102:1111 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 2 57 theme acidic 249:254 arg1 propeptide 269:278 an acidic 2-amino acid propeptide 246:278 an acidic 2-amino acid propeptide 246:278 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 10 58 from amplification 1706:1718 arg1 libraries 1773:1781 placental cDNA libraries 1758:1781 placental cDNA libraries 1758:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 10 58 from amplification 1706:1718 arg1 skin 1749:1752 skin 1749:1752 skin 1749:1752 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 9 59 theme chymase 1523:1529 arg1 DNA 1531:1533 chymase DNA 1523:1533 chymase DNA 1523:1533 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 5 60 theme cell 789:792 arg1 chymase 794:800 dog mast cell chymase 780:800 dog mast cell chymase (83%) 780:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 60 theme cell 789:792 arg1 % 805:805 83% 803:805 83% 803:805 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 4 61 theme NH2-terminal 497:508 arg1 sequence 521:528 NH2-terminal amino acid sequence 497:528 NH2-terminal amino acid sequence reported for human skin chymase 497:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 7 62 theme serine 1266:1271 arg1 proteases 1273:1281 the major mast cell serine proteases 1246:1281 the major mast cell serine proteases 1246:1281 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 7 63 theme major 1250:1254 arg1 proteases 1273:1281 the major mast cell serine proteases 1246:1281 the major mast cell serine proteases 1246:1281 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 10 64 theme cDNA 1768:1771 arg1 libraries 1773:1781 placental cDNA libraries 1758:1781 placental cDNA libraries 1758:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 6 65 theme lymphocyte 1076:1085 arg1 granzymes 1087:1095 lymphocyte granzymes 1076:1095 lymphocyte granzymes 1076:1095 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 5 66 theme identity 768:775 arg1 level 820:824 a lower level 812:824 a lower level of identity to the sequences of rodent chymases (58-62%) 812:881 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 66 theme identity 768:775 arg1 level 750:754 a high level 743:754 a high level of sequence identity to dog mast cell chymase (83%) 743:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 9 67 theme human 1540:1544 arg1 blot 1558:1561 a human genomic DNA blot 1538:1561 a human genomic DNA blot 1538:1561 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 2 68 theme acid 264:267 arg1 propeptide 269:278 an acidic 2-amino acid propeptide 246:278 an acidic 2-amino acid propeptide 246:278 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 9 69 theme DNA 1554:1556 arg1 blot 1558:1561 a human genomic DNA blot 1538:1561 a human genomic DNA blot 1538:1561 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 7 70 theme additional 1221:1230 arg1 evidence 1232:1239 additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance 1221:1332 additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance 1221:1332 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 4 71 theme human 543:547 arg1 chymase 554:560 human skin chymase 543:560 human skin chymase 543:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 5 72 theme high 745:748 arg1 level 750:754 a high level 743:754 a high level of sequence identity to dog mast cell chymase (83%) 743:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 10 73 theme chymase 1643:1649 arg1 gene 1651:1654 the chymase gene 1639:1654 the chymase gene 1639:1654 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 7 74 theme cell 1196:1199 arg1 tryptase 1201:1208 mast cell tryptase 1191:1208 mast cell tryptase 1191:1208 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 4 75 theme NH2-terminal 603:614 arg1 sequence 640:647 NH2-terminal and internal amino acid sequence 603:647 NH2-terminal and internal amino acid sequence reported for human heart chymase 603:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 2 76 theme signal 230:235 arg1 peptide 237:243 a 19-amino acid signal peptide 214:243 a 19-amino acid signal peptide 214:243 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 4 77 theme internal 620:627 arg1 sequence 640:647 NH2-terminal and internal amino acid sequence 603:647 NH2-terminal and internal amino acid sequence reported for human heart chymase 603:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 2 78 theme 19-amino 216:223 arg1 peptide 237:243 a 19-amino acid signal peptide 214:243 a 19-amino acid signal peptide 214:243 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 5 79 theme dog 780:782 arg1 chymase 794:800 dog mast cell chymase 780:800 dog mast cell chymase (83%) 780:806 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 79 theme dog 780:782 arg1 % 805:805 83% 803:805 83% 803:805 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 6 80 theme leukocyte 1022:1030 arg1 proteases 1039:1047 several other granule-associated leukocyte serine proteases 989:1047 several other granule-associated leukocyte serine proteases 989:1047 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 80 theme leukocyte 1022:1030 arg1 granzymes 1087:1095 lymphocyte granzymes 1076:1095 lymphocyte granzymes 1076:1095 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 80 theme leukocyte 1022:1030 arg1 chymase 1064:1070 rat chymase II 1060:1073 rat chymase II 1060:1073 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 80 theme leukocyte 1022:1030 arg1 G 1123:1123 neutrophil cathespin G 1102:1123 neutrophil cathespin G 1102:1123 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 8 81 theme gene 1461:1464 arg1 localization 1433:1444 localization 1433:1444 localization of the chymase gene to human chromosome 14 1433:1487 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 1 82 theme human 122:126 arg1 chymase 128:134 human chymase 122:134 human chymase 122:134 A gene encoding human chymase was cloned and sequenced. 2071582 9 83 theme chymase 1611:1617 arg1 gene 1619:1622 more than one human chymase gene 1591:1622 more than one human chymase gene 1591:1622 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 8 84 theme chymase 1352:1358 arg1 fragments 1374:1382 chymase gene-specific fragments 1352:1382 chymase gene-specific fragments from hamster/human hybrid cell line DNA 1352:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 0 85 theme gene 78:81 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 0 85 theme gene 78:81 arg1 assignment 23:32 chromosomal assignment 11:32 chromosomal assignment 11:32 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 0 85 theme gene 78:81 arg1 sequence 58:65 deduced amino acid sequence 39:65 deduced amino acid sequence of a human gene for mast cell chymase 39:103 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 8 86 theme fragments 1374:1382 arg1 Amplification 1335:1347 Amplification 1335:1347 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA 1335:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 4 87 theme human 662:666 arg1 chymase 674:680 human heart chymase 662:680 human heart chymase 662:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 0 88 theme mast 87:90 arg1 chymase 97:103 mast cell chymase 87:103 mast cell chymase 87:103 Structure, chromosomal assignment, and deduced amino acid sequence of a human gene for mast cell chymase. 2071582 7 89 theme gene 1152:1155 arg1 organization 1157:1168 the gene organization 1148:1168 the gene organization 1148:1168 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 10 90 theme chymase-specific 1723:1738 arg1 DNA 1740:1742 chymase-specific DNA 1723:1742 chymase-specific DNA from skin and placental cDNA libraries 1723:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 8 91 theme hamster/human 1389:1401 arg1 line 1415:1418 hamster/human hybrid cell line 1389:1418 hamster/human hybrid cell line DNA 1389:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 4 92 theme acid 516:519 arg1 sequence 521:528 NH2-terminal amino acid sequence 497:528 NH2-terminal amino acid sequence reported for human skin chymase 497:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 3 93 theme +13 381:383 arg1 charge 371:376 net charge 367:376 net charge of +13 367:383 The mature enzyme is predicted to be cationic (net charge of +13) and to be modified by N-glycosylation at two sites. 2071582 8 94 theme cell 1410:1413 arg1 line 1415:1418 hamster/human hybrid cell line 1389:1418 hamster/human hybrid cell line DNA 1389:1422 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 4 95 theme acid 635:638 arg1 sequence 640:647 NH2-terminal and internal amino acid sequence 603:647 NH2-terminal and internal amino acid sequence reported for human heart chymase 603:680 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 6 96 from similar 970:976 arg1 placement 901:909 placement 901:909 placement 901:909 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 96 from similar 970:976 arg1 phase 891:895 phase 891:895 phase 891:895 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 5 97 theme catalytic 718:726 arg1 domain 728:733 the catalytic domain 714:733 the catalytic domain 714:733 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 98 theme lower 814:818 arg1 level 820:824 a lower level 812:824 a lower level of identity to the sequences of rodent chymases (58-62%) 812:881 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 7 99 theme cell 1261:1264 arg1 proteases 1273:1281 the major mast cell serine proteases 1246:1281 the major mast cell serine proteases 1246:1281 However, the gene organization differs from that of mast cell tryptase, providing additional evidence that the major mast cell serine proteases are separated by substantial evolutionary distance. 2071582 6 100 theme introns 914:920 arg1 placement 901:909 placement 901:909 placement 901:909 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 100 theme introns 914:920 arg1 phase 891:895 phase 891:895 phase 891:895 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 4 101 theme acid 448:451 arg1 sequence 453:460 The amino acid sequence 438:460 The amino acid sequence 438:460 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 4 101 theme acid 448:451 arg1 identical 569:577 identical 569:577 identical 569:577 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 4 101 theme acid 448:451 arg1 identical 465:473 identical 465:473 identical 465:473 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 5 102 theme predicted 692:700 arg1 sequence 702:709 The full predicted sequence 683:709 The full predicted sequence of the catalytic domain 683:733 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 6 103 theme gene 962:965 arg1 organization 927:938 the organization 923:938 the organization of this human chymase gene 923:965 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 6 103 theme gene 962:965 arg1 similar 970:976 similar 970:976 similar 970:976 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 10 104 theme DNA 1740:1742 arg1 amplification 1706:1718 the amplification 1702:1718 the amplification of chymase-specific DNA from skin and placental cDNA libraries 1702:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 2071582 2 105 theme 226-amino 287:295 arg1 domain 312:317 a 226-amino acid catalytic domain 285:317 a 226-amino acid catalytic domain 285:317 The protein-coding exons reveal a preproenzyme with a 19-amino acid signal peptide, an acidic 2-amino acid propeptide, and a 226-amino acid catalytic domain. 2071582 9 106 theme High 1490:1493 arg1 hybridization 1506:1518 High stringency hybridization 1490:1518 High stringency hybridization of chymase DNA to a human genomic DNA blot 1490:1561 High stringency hybridization of chymase DNA to a human genomic DNA blot suggests the possibility of more than one human chymase gene. 2071582 6 107 theme human 948:952 arg1 gene 962:965 this human chymase gene 943:965 this human chymase gene 943:965 In the phase and placement of introns, the organization of this human chymase gene is similar to that of several other granule-associated leukocyte serine proteases, including rat chymase II, lymphocyte granzymes, and neutrophil cathespin G and elastase. 2071582 4 108 theme sequence 521:528 arg1 sequence 521:528 NH2-terminal amino acid sequence 497:528 NH2-terminal amino acid sequence reported for human skin chymase 497:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 4 108 theme sequence 521:528 arg1 residues 485:492 the 35 residues 478:492 the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase 478:560 The amino acid sequence is identical to the 35 residues of NH2-terminal amino acid sequence reported for human skin chymase and is identical to 29 of 31 residues of NH2-terminal and internal amino acid sequence reported for human heart chymase. 2071582 5 109 theme rodent 858:863 arg1 chymases 865:872 rodent chymases 858:872 rodent chymases (58-62%) 858:881 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 5 109 theme rodent 858:863 arg1 % 880:880 58-62% 875:880 58-62% 875:880 The full predicted sequence of the catalytic domain reveals a high level of sequence identity to dog mast cell chymase (83%) and a lower level of identity to the sequences of rodent chymases (58-62%). 2071582 8 110 theme human 1469:1473 arg1 chromosome 1475:1484 human chromosome 14 1469:1487 human chromosome 14 1469:1487 Amplification of chymase gene-specific fragments from hamster/human hybrid cell line DNA suggests localization of the chymase gene to human chromosome 14. 2071582 10 111 theme placental 1758:1766 arg1 libraries 1773:1781 placental cDNA libraries 1758:1781 placental cDNA libraries 1758:1781 Evidence that the chymase gene is expressed in human tissues was obtained by the amplification of chymase-specific DNA from skin and placental cDNA libraries. 6267033 10 0 theme human 949:953 arg1 peptide 955:961 the human peptide 945:961 the human peptide 945:961 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 5 1 theme genomic 560:566 arg1 sequence 572:579 the reported genomic DNA sequence 547:579 the reported genomic DNA sequence 547:579 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 11 2 theme adrenal 1126:1132 arg1 tumor 1134:1138 a human adrenal tumor 1118:1138 a human adrenal tumor 1118:1138 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 12 3 theme amino 1382:1386 arg1 acids 1388:1392 the first 19 amino acids 1369:1392 the first 19 amino acids due to the presence of a 2-kilobase intron 1369:1435 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 9 4 dep 8 776:776 arg1 to 773:774 to 773:774 to 773:774 Two disulfide bridges linking Cys 2 to 8 and Cys 20 to 24 have been determined. 6267033 4 5 gly O-glycosylation 465:479 arg2 Thr 489:491 Thr 45 489:494 Thr 45 489:494 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 5 gly O-glycosylation 465:479 arg2 site 481:484 an O-glycosylation site 462:484 an O-glycosylation site at Thr 45 462:494 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 5 gly O-glycosylation 465:479 arg2 Asn 527:529 Asn 65 527:532 Asn 65 527:532 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 2 6 gly glycopeptide 196:207 arg2 glycopeptide 196:207 a human glycopeptide 188:207 a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin 188:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 11 7 from cells 1109:1113 arg1 release 1072:1078 the release 1068:1078 the release of aldosterone from isolated cells of a human adrenal tumor 1068:1138 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 4 8 theme N-glycosidic 503:514 arg1 linkage 516:522 an N-glycosidic linkage 500:522 an N-glycosidic linkage at Asn 65 500:532 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 9 from Asn 527:529 arg1 linkage 516:522 an N-glycosidic linkage 500:522 an N-glycosidic linkage at Asn 65 500:532 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 9 from Asn 527:529 arg1 site 481:484 an O-glycosylation site 462:484 an O-glycosylation site at Thr 45 462:494 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 0 10 gly glycopeptide 81:92 arg2 glycopeptide 81:92 pro-opiomelanocortin NH2-terminal glycopeptide 47:92 pro-opiomelanocortin NH2-terminal glycopeptide 47:92 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 11 11 theme adrenocorticotropic 1168:1186 arg1 hormone 1188:1194 adrenocorticotropic hormone 1168:1194 adrenocorticotropic hormone 1168:1194 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 5 12 dep Chang 582:586 arg1 N 625:625 N 625:625 N 625:625 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 12 13 theme human 1341:1345 arg1 DNA 1347:1349 the human DNA 1337:1349 the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron 1337:1435 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 12 14 theme reported 1301:1308 arg1 sequence 1310:1317 this reported sequence 1296:1317 this reported sequence 1296:1317 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 9 15 theme disulfide 741:749 arg1 bridges 751:757 Two disulfide bridges 737:757 Two disulfide bridges linking Cys 2 to 8 and Cys 20 to 24 737:793 Two disulfide bridges linking Cys 2 to 8 and Cys 20 to 24 have been determined. 6267033 3 16 theme amino 435:439 arg1 acids 441:445 76 amino acids 432:445 76 amino acids 432:445 The complete sequence of this peptide was determined following CNBr fragmentation and it is shown to be 76 amino acids long. 6267033 11 17 theme NH2-terminal 1025:1036 arg1 peptide 1038:1044 The human NH2-terminal peptide 1015:1044 The human NH2-terminal peptide 1015:1044 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 11 18 theme porcine 1204:1210 arg1 times 1248:1252 100 times 1244:1252 100 times more potent than angiotensin II 1244:1284 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 11 18 theme porcine 1204:1210 arg1 analogue 1225:1232 the porcine NH2-terminal analogue 1200:1232 the porcine NH2-terminal analogue 1200:1232 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 2 19 theme complete 163:170 arg1 purification 172:183 complete purification 163:183 complete purification 163:183 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 4 20 contain bears 456:460 arg2 linkage 516:522 an N-glycosidic linkage 500:522 an N-glycosidic linkage at Asn 65 500:532 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 20 contain bears 456:460 arg2 site 481:484 an O-glycosylation site 462:484 an O-glycosylation site at Thr 45 462:494 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 20 contain bears 456:460 arg1 It 453:454 It 453:454 It 453:454 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 0 21 theme Primary 0:6 arg1 structure 8:16 Primary structure 0:16 Primary structure of the major human pituitary 0:45 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 4 22 theme O-glycosylation 465:479 arg1 site 481:484 an O-glycosylation site 462:484 an O-glycosylation site at Thr 45 462:494 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 11 23 theme NH2-terminal 1212:1223 arg1 times 1248:1252 100 times 1244:1252 100 times more potent than angiotensin II 1244:1284 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 11 23 theme NH2-terminal 1212:1223 arg1 analogue 1225:1232 the porcine NH2-terminal analogue 1200:1232 the porcine NH2-terminal analogue 1200:1232 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 11 24 theme aldosterone 1083:1093 arg1 release 1072:1078 the release 1068:1078 the release of aldosterone from isolated cells of a human adrenal tumor 1068:1138 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 3 25 theme complete 332:339 arg1 sequence 341:348 The complete sequence 328:348 The complete sequence of this peptide 328:364 The complete sequence of this peptide was determined following CNBr fragmentation and it is shown to be 76 amino acids long. 6267033 1 26 theme aldosterone-stimulating 111:133 arg1 activity 135:142 an aldosterone-stimulating activity 108:142 an aldosterone-stimulating activity 108:142 Evidence for an aldosterone-stimulating activity. 6267033 10 27 theme bridge 853:858 arg1 localization 860:871 the sequence and disulfide bridge localization 826:871 localization 860:871 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 10 28 theme porcine 995:1001 arg1 calcitonin 1003:1012 porcine calcitonin 995:1012 porcine calcitonin 995:1012 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 10 28 theme porcine 995:1001 arg1 calcitonins 971:981 all calcitonins 967:981 all calcitonins 967:981 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 11 29 theme isolated 1100:1107 arg1 cells 1109:1113 isolated cells 1100:1113 isolated cells of a human adrenal tumor 1100:1138 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 0 30 theme human 31:35 arg1 pituitary 37:45 the major human pituitary 21:45 the major human pituitary 21:45 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 4 31 from Thr 489:491 arg1 linkage 516:522 an N-glycosidic linkage 500:522 an N-glycosidic linkage at Asn 65 500:532 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 4 31 from Thr 489:491 arg1 site 481:484 an O-glycosylation site 462:484 an O-glycosylation site at Thr 45 462:494 It bears an O-glycosylation site at Thr 45 and an N-glycosidic linkage at Asn 65. 6267033 10 32 theme homology 892:899 arg1 degree 882:887 a large degree 874:887 a large degree of homology 874:899 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 2 33 theme segment 282:288 arg1 form 247:250 the major immunoreactive form 222:250 the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin 222:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 0 34 theme major 25:29 arg1 pituitary 37:45 the major human pituitary 21:45 the major human pituitary 21:45 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 12 35 theme 2-kilobase 1419:1428 arg1 intron 1430:1435 a 2-kilobase intron 1417:1435 a 2-kilobase intron 1417:1435 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 9 36 dep 24 792:793 arg1 to 789:790 to 789:790 to 789:790 Two disulfide bridges linking Cys 2 to 8 and Cys 20 to 24 have been determined. 6267033 2 37 theme glycopeptide 196:207 arg1 isolation 149:157 isolation 149:157 isolation 149:157 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 2 37 theme glycopeptide 196:207 arg1 purification 172:183 complete purification 163:183 complete purification 163:183 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 2 38 theme NH2-terminal 269:280 arg1 segment 282:288 the pituitary NH2-terminal segment 255:288 the pituitary NH2-terminal segment of pro-opiomelanocortin 255:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 5 39 theme DNA 568:570 arg1 sequence 572:579 the reported genomic DNA sequence 547:579 the reported genomic DNA sequence 547:579 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 12 40 theme intron 1430:1435 arg1 presence 1405:1412 the presence 1401:1412 the presence of a 2-kilobase intron 1401:1435 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 0 41 theme pro-opiomelanocortin 47:66 arg1 glycopeptide 81:92 pro-opiomelanocortin NH2-terminal glycopeptide 47:92 pro-opiomelanocortin NH2-terminal glycopeptide 47:92 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 3 42 theme peptide 358:364 arg1 sequence 341:348 The complete sequence 328:348 The complete sequence of this peptide 328:364 The complete sequence of this peptide was determined following CNBr fragmentation and it is shown to be 76 amino acids long. 6267033 2 43 theme human 190:194 arg1 glycopeptide 196:207 a human glycopeptide 188:207 a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin 188:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 2 44 theme pituitary 259:267 arg1 segment 282:288 the pituitary NH2-terminal segment 255:288 the pituitary NH2-terminal segment of pro-opiomelanocortin 255:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 5 45 dep Compared 535:542 arg1 Y. 595:596 Y. 595:596 Y. 595:596 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 5 45 dep Compared 535:542 arg1 1980 629:632 1980 629:632 1980 629:632 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 5 45 dep Compared 535:542 arg1 Chang 582:586 Chang 582:586 Chang 582:586 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 5 45 dep Compared 535:542 arg1 Proc 635:638 Proc 635:638 Proc 635:638 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 5 45 dep Compared 535:542 arg1 S. 622:623 S. 622:623 S. 622:623 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 5 45 dep Compared 535:542 arg1 M. 607:608 M. 607:608 M. 607:608 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 6267033 12 46 theme due 1394:1396 arg1 acids 1388:1392 the first 19 amino acids 1369:1392 the first 19 amino acids due to the presence of a 2-kilobase intron 1369:1435 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 0 47 theme pituitary 37:45 arg1 structure 8:16 Primary structure 0:16 Primary structure of the major human pituitary 0:45 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 11 48 theme tumor 1134:1138 arg1 cells 1109:1113 isolated cells 1100:1113 isolated cells of a human adrenal tumor 1100:1138 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 8 49 dep A. 664:665 arg1 variation 687:695 one variation 683:695 one variation 683:695 U. S. A. 77, 4890-4894), one variation exists, namely Arg 22 replacing Gly 22. 6267033 12 50 theme first 1373:1377 arg1 acids 1388:1392 the first 19 amino acids 1369:1392 the first 19 amino acids due to the presence of a 2-kilobase intron 1369:1435 Finally, this reported sequence completes that of the human DNA which was lacking the first 19 amino acids due to the presence of a 2-kilobase intron. 6267033 2 51 dep isolation 149:157 arg1 The 145:147 The 145:147 The 145:147 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 10 52 theme NH2-terminal 920:931 arg1 sequence 933:940 the NH2-terminal sequence 916:940 the NH2-terminal sequence of the human peptide 916:961 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 10 53 theme large 876:880 arg1 degree 882:887 a large degree 874:887 a large degree of homology 874:899 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 10 54 theme disulfide 843:851 arg1 localization 860:871 the sequence and disulfide bridge localization 826:871 localization 860:871 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 0 55 dep glycopeptide 81:92 arg1 structure 8:16 Primary structure 0:16 Primary structure of the major human pituitary 0:45 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 11 56 theme human 1120:1124 arg1 tumor 1134:1138 a human adrenal tumor 1118:1138 a human adrenal tumor 1118:1138 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 2 57 theme immunoreactive 232:245 arg1 form 247:250 the major immunoreactive form 222:250 the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin 222:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 11 58 theme potent 1259:1264 arg1 times 1248:1252 100 times 1244:1252 100 times more potent than angiotensin II 1244:1284 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 11 58 theme potent 1259:1264 arg1 analogue 1225:1232 the porcine NH2-terminal analogue 1200:1232 the porcine NH2-terminal analogue 1200:1232 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 3 59 theme CNBr 391:394 arg1 fragmentation 396:408 CNBr fragmentation 391:408 CNBr fragmentation 391:408 The complete sequence of this peptide was determined following CNBr fragmentation and it is shown to be 76 amino acids long. 6267033 2 60 theme major 226:230 arg1 form 247:250 the major immunoreactive form 222:250 the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin 222:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 2 61 theme pro-opiomelanocortin 293:312 arg1 segment 282:288 the pituitary NH2-terminal segment 255:288 the pituitary NH2-terminal segment of pro-opiomelanocortin 255:312 The isolation and complete purification of a human glycopeptide representing the major immunoreactive form of the pituitary NH2-terminal segment of pro-opiomelanocortin is presented. 6267033 0 62 theme NH2-terminal 68:79 arg1 glycopeptide 81:92 pro-opiomelanocortin NH2-terminal glycopeptide 47:92 pro-opiomelanocortin NH2-terminal glycopeptide 47:92 Primary structure of the major human pituitary pro-opiomelanocortin NH2-terminal glycopeptide. 6267033 10 63 theme peptide 955:961 arg1 calcitonin 1003:1012 porcine calcitonin 995:1012 porcine calcitonin 995:1012 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 10 63 theme peptide 955:961 arg1 calcitonins 971:981 all calcitonins 967:981 all calcitonins 967:981 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 10 63 theme peptide 955:961 arg1 sequence 933:940 the NH2-terminal sequence 916:940 the NH2-terminal sequence of the human peptide 916:961 Based on the sequence and disulfide bridge localization, a large degree of homology exists between the NH2-terminal sequence of the human peptide and all calcitonins, especially porcine calcitonin. 6267033 11 64 theme human 1019:1023 arg1 peptide 1038:1044 The human NH2-terminal peptide 1015:1044 The human NH2-terminal peptide 1015:1044 The human NH2-terminal peptide is shown to stimulate the release of aldosterone from isolated cells of a human adrenal tumor, in at least equipotency to adrenocorticotropic hormone and the porcine NH2-terminal analogue, which is 100 times more potent than angiotensin II. 6267033 5 65 theme reported 551:558 arg1 sequence 572:579 the reported genomic DNA sequence 547:579 the reported genomic DNA sequence 547:579 Compared to the reported genomic DNA sequence (Chang, A. C. Y., Cochet, M., and Cohen, S. N. (1980) Proc. 9722584 8 0 theme precursor 1092:1100 arg1 protein 1102:1108 the precursor protein 1088:1108 the precursor protein 1088:1108 In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. 9722584 7 1 theme porcine 983:989 arg1 NTPPHase 991:998 porcine NTPPHase 983:998 porcine NTPPHase 983:998 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 5 2 theme post-translational 666:683 arg1 modifications 685:697 post-translational modifications 666:697 post-translational modifications 666:697 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 1 3 theme protein 218:224 arg1 expression 163:172 expression 163:172 expression 163:172 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 3 theme protein 218:224 arg1 cloning 151:157 cDNA cloning 146:157 cDNA cloning 146:157 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 0 4 theme protein 61:67 arg1 sequence 31:38 deduced amino acid sequence 12:38 deduced amino acid sequence 12:38 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 0 4 theme protein 61:67 arg1 Cloning 0:6 Cloning 0:6 Cloning 0:6 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 5 5 theme carboxyl-terminal 708:724 arg1 half 726:729 the carboxyl-terminal half 704:729 the carboxyl-terminal half 704:729 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 5 6 theme amino-terminal 583:596 arg1 half 598:601 The amino-terminal half 579:601 The amino-terminal half 579:601 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 7 7 gly N-glycosylation 1062:1076 arg2 sites 1078:1082 two putative N-glycosylation sites 1049:1082 two putative N-glycosylation sites 1049:1082 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 7 7 gly N-glycosylation 1062:1076 arg2 two 1049:1051 two 1049:1051 two 1049:1051 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 1 8 theme cDNA 146:149 arg1 cloning 151:157 cDNA cloning 146:157 cDNA cloning 146:157 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 9 9 theme furin-like 1445:1454 arg1 protease 1456:1463 a furin-like protease 1443:1463 a furin-like protease 1443:1463 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 6 10 theme N-glycosylation 939:953 arg1 sites 955:959 six putative N-glycosylation sites 926:959 six putative N-glycosylation sites 926:959 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 7 11 theme N-glycosylation 1062:1076 arg1 sites 1078:1082 two putative N-glycosylation sites 1049:1082 two putative N-glycosylation sites 1049:1082 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 4 12 contain has 492:494 arg2 peptide 514:520 a putative signal peptide 496:520 a putative signal peptide of 21 amino acids 496:538 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 4 12 contain has 492:494 arg1 protein 484:490 The protein 480:490 The protein 480:490 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 6 13 theme putative 930:937 arg1 sites 955:959 six putative N-glycosylation sites 926:959 six putative N-glycosylation sites 926:959 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 8 14 theme NTPPHase 1114:1121 arg1 region 1123:1128 the NTPPHase region 1110:1128 the NTPPHase region 1110:1128 In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. 9722584 9 15 theme cell-free 1274:1282 arg1 system 1296:1301 a cell-free translation system 1272:1301 a cell-free translation system 1272:1301 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 1 16 theme human 240:244 arg1 cartilage 256:264 human articular cartilage 240:264 human articular cartilage 240:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 5 17 dep CILP 618:621 arg1 including 656:664 including 656:664 not including post-translational modifications 652:697 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 9 18 theme polypeptide 1394:1404 arg1 chain 1406:1410 a single polypeptide chain 1385:1410 a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion 1385:1514 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 9 18 theme polypeptide 1394:1404 arg1 protein 1359:1365 the precursor protein 1345:1365 the precursor protein 1345:1365 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 8 19 theme cleavage 1205:1212 arg1 sequence 1224:1231 a furin proteinase cleavage consensus sequence 1186:1231 a furin proteinase cleavage consensus sequence 1186:1231 In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. 9722584 4 20 theme acids 534:538 arg1 peptide 514:520 a putative signal peptide 496:520 a putative signal peptide of 21 amino acids 496:538 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 7 21 theme human 966:970 arg1 homolog 972:978 The human homolog 962:978 The human homolog of porcine NTPPHase described here 962:1013 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 4 22 theme amino 528:532 arg1 acids 534:538 21 amino acids 525:538 21 amino acids 525:538 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 8 23 theme proteinase 1194:1203 arg1 sequence 1224:1231 a furin proteinase cleavage consensus sequence 1186:1231 a furin proteinase cleavage consensus sequence 1186:1231 In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. 9722584 9 24 theme EBNA 1319:1322 arg1 cells 1324:1328 COS-7 or EBNA cells 1310:1328 cells 1324:1328 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 4 25 theme signal 507:512 arg1 peptide 514:520 a putative signal peptide 496:520 a putative signal peptide of 21 amino acids 496:538 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 9 26 theme translation 1284:1294 arg1 system 1296:1301 a cell-free translation system 1272:1301 a cell-free translation system 1272:1301 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 9 27 from Expression 1234:1243 arg1 system 1296:1301 a cell-free translation system 1272:1301 a cell-free translation system 1272:1301 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 9 27 from Expression 1234:1243 arg1 cells 1324:1328 COS-7 or EBNA cells 1310:1328 cells 1324:1328 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 7 28 theme putative 1053:1060 arg1 sites 1078:1082 two putative N-glycosylation sites 1049:1082 two putative N-glycosylation sites 1049:1082 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 1 29 theme articular 246:254 arg1 cartilage 256:264 human articular cartilage 240:264 human articular cartilage 240:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 9 30 theme single 1387:1392 arg1 chain 1406:1410 a single polypeptide chain 1385:1410 a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion 1385:1514 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 9 30 theme single 1387:1392 arg1 protein 1359:1365 the precursor protein 1345:1365 the precursor protein 1345:1365 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 7 31 contain contains 1015:1022 arg2 residues 1036:1043 10 cysteine residues 1024:1043 10 cysteine residues 1024:1043 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 7 31 contain contains 1015:1022 arg2 sites 1078:1082 two putative N-glycosylation sites 1049:1082 two putative N-glycosylation sites 1049:1082 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 7 31 contain contains 1015:1022 arg1 homolog 972:978 The human homolog 962:978 The human homolog of porcine NTPPHase described here 962:1013 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 5 32 theme molecular 823:831 arg1 mass 833:836 molecular mass 823:836 molecular mass of 51.8 kDa 823:848 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 0 33 theme amino 20:24 arg1 sequence 31:38 deduced amino acid sequence 12:38 deduced amino acid sequence 12:38 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 4 34 theme putative 498:505 arg1 peptide 514:520 a putative signal peptide 496:520 a putative signal peptide of 21 amino acids 496:538 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 5 35 theme post-translational 865:882 arg1 modifications 884:896 post-translational modifications 865:896 post-translational modifications 865:896 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 6 36 contain has 905:907 arg1 CILP 900:903 CILP 900:903 CILP 900:903 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 6 36 contain has 905:907 arg2 sites 955:959 six putative N-glycosylation sites 926:959 six putative N-glycosylation sites 926:959 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 6 36 contain has 905:907 arg2 cysteines 912:920 30 cysteines 909:920 30 cysteines 909:920 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 3 37 theme acids 427:431 arg1 polypeptide 401:411 a polypeptide 399:411 a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa 399:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 3 37 theme acids 427:431 arg1 mRNA 348:351 2-kilobase mRNA 337:351 2-kilobase mRNA detected in human articular cartilage 337:389 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 0 38 theme deduced 12:18 arg1 sequence 31:38 deduced amino acid sequence 12:38 deduced amino acid sequence 12:38 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 7 39 theme cysteine 1027:1034 arg1 residues 1036:1043 10 cysteine residues 1024:1043 10 cysteine residues 1024:1043 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 5 40 theme kDa 846:848 arg1 mass 833:836 molecular mass 823:836 molecular mass of 51.8 kDa 823:848 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 5 41 theme porcine 772:778 arg1 NTPPHase 813:820 NTPPHase 813:820 NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications) 813:897 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 5 41 theme porcine 772:778 arg1 pyrophosphohydrolase 791:810 a porcine nucleotide pyrophosphohydrolase 770:810 a porcine nucleotide pyrophosphohydrolase 770:810 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 3 42 theme articular 371:379 arg1 cartilage 381:389 human articular cartilage 365:389 human articular cartilage 365:389 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 1 43 theme cartilage 267:275 arg1 CILP 305:308 CILP 305:308 CILP 305:308 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 43 theme cartilage 267:275 arg1 protein 296:302 cartilage intermediate layer protein 267:302 cartilage intermediate layer protein (CILP) 267:309 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 43 theme cartilage 267:275 arg1 protein 218:224 a novel protein 210:224 a novel protein isolated from human articular cartilage 210:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 0 44 theme nucleotide 109:118 arg1 pyrophosphohydrolase 120:139 a nucleotide pyrophosphohydrolase 107:139 a nucleotide pyrophosphohydrolase 107:139 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 5 45 theme nucleotide 780:789 arg1 NTPPHase 813:820 NTPPHase 813:820 NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications) 813:897 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 5 45 theme nucleotide 780:789 arg1 pyrophosphohydrolase 791:810 a porcine nucleotide pyrophosphohydrolase 770:810 a porcine nucleotide pyrophosphohydrolase 770:810 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 4 46 theme polypeptides 565:576 arg1 proform 550:556 a proform 548:556 a proform of two polypeptides 548:576 The protein has a putative signal peptide of 21 amino acids, and is a proform of two polypeptides. 9722584 8 47 theme consensus 1214:1222 arg1 sequence 1224:1231 a furin proteinase cleavage consensus sequence 1186:1231 a furin proteinase cleavage consensus sequence 1186:1231 In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. 9722584 3 48 theme 1184 416:419 arg1 acids 427:431 1184 amino acids 416:431 1184 amino acids 416:431 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 0 49 theme acid 26:29 arg1 sequence 31:38 deduced amino acid sequence 12:38 deduced amino acid sequence 12:38 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 1 50 theme intermediate 277:288 arg1 CILP 305:308 CILP 305:308 CILP 305:308 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 50 theme intermediate 277:288 arg1 protein 296:302 cartilage intermediate layer protein 267:302 cartilage intermediate layer protein (CILP) 267:309 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 50 theme intermediate 277:288 arg1 protein 218:224 a novel protein 210:224 a novel protein isolated from human articular cartilage 210:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 5 51 theme kDa 647:649 arg1 mass 634:637 molecular mass 624:637 molecular mass of 78.5 kDa 624:649 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 5 52 theme homologous 756:765 arg1 protein 748:754 a protein 746:754 a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications) 746:897 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 6 53 gly N-glycosylation 939:953 arg2 six 926:928 six 926:928 six 926:928 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 6 53 gly N-glycosylation 939:953 arg2 sites 955:959 six putative N-glycosylation sites 926:959 six putative N-glycosylation sites 926:959 CILP has 30 cysteines and six putative N-glycosylation sites. 9722584 3 54 theme amino 421:425 arg1 acids 427:431 1184 amino acids 416:431 1184 amino acids 416:431 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 8 55 theme furin 1188:1192 arg1 sequence 1224:1231 a furin proteinase cleavage consensus sequence 1186:1231 a furin proteinase cleavage consensus sequence 1186:1231 In the precursor protein the NTPPHase region is immediately preceded by a tetrapeptide conforming to a furin proteinase cleavage consensus sequence. 9722584 1 56 theme layer 290:294 arg1 CILP 305:308 CILP 305:308 CILP 305:308 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 56 theme layer 290:294 arg1 protein 296:302 cartilage intermediate layer protein 267:302 cartilage intermediate layer protein (CILP) 267:309 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 56 theme layer 290:294 arg1 protein 218:224 a novel protein 210:224 a novel protein isolated from human articular cartilage 210:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 5 57 dep including 656:664 arg1 mass 634:637 molecular mass 624:637 molecular mass of 78.5 kDa 624:649 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 9 58 theme COS-7 1310:1314 arg1 cells 1324:1328 COS-7 or EBNA cells 1310:1328 cells 1324:1328 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 3 59 theme molecular 451:459 arg1 mass 461:464 a calculated molecular mass 438:464 a calculated molecular mass of 132.5 kDa 438:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 3 60 theme human 365:369 arg1 cartilage 381:389 human articular cartilage 365:389 human articular cartilage 365:389 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 3 61 located detected 353:360 arg2 polypeptide 401:411 a polypeptide 399:411 a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa 399:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 3 61 located detected 353:360 arg2 mRNA 348:351 2-kilobase mRNA 337:351 2-kilobase mRNA detected in human articular cartilage 337:389 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 3 61 located detected 353:360 arg1 cartilage 381:389 human articular cartilage 365:389 human articular cartilage 365:389 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 5 62 dep NTPPHase 813:820 arg1 including 855:863 including 855:863 not including post-translational modifications 851:896 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 9 63 theme cDNA 1264:1267 arg1 Expression 1234:1243 Expression 1234:1243 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells 1234:1328 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 3 64 with polypeptide 401:411 arg1 mass 461:464 a calculated molecular mass 438:464 a calculated molecular mass of 132.5 kDa 438:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 1 65 attach isolated 226:233 arg2 protein 296:302 cartilage intermediate layer protein 267:302 cartilage intermediate layer protein (CILP) 267:309 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 65 attach isolated 226:233 arg2 protein 218:224 a novel protein 210:224 a novel protein isolated from human articular cartilage 210:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 65 attach isolated 226:233 arg1 cartilage 256:264 human articular cartilage 240:264 human articular cartilage 240:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 9 66 theme precursor 1349:1357 arg1 chain 1406:1410 a single polypeptide chain 1385:1410 a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion 1385:1514 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 9 66 theme precursor 1349:1357 arg1 protein 1359:1365 the precursor protein 1345:1365 the precursor protein 1345:1365 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 5 67 theme molecular 624:632 arg1 mass 634:637 molecular mass 624:637 molecular mass of 78.5 kDa 624:649 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 9 68 theme preceding 1496:1504 arg1 secretion 1506:1514 or preceding secretion 1493:1514 secretion 1506:1514 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 5 69 dep including 855:863 arg1 mass 833:836 molecular mass 823:836 molecular mass of 51.8 kDa 823:848 The amino-terminal half corresponds to CILP (molecular mass of 78.5 kDa, not including post-translational modifications) and the carboxyl-terminal half corresponds to a protein homologous to a porcine nucleotide pyrophosphohydrolase, NTPPHase (molecular mass of 51.8 kDa, not including post-translational modifications). 9722584 3 70 theme calculated 440:449 arg1 mass 461:464 a calculated molecular mass 438:464 a calculated molecular mass of 132.5 kDa 438:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 0 71 theme cartilage 51:59 arg1 protein 61:67 a novel cartilage protein 43:67 a novel cartilage protein (CILP) 43:74 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 0 71 theme cartilage 51:59 arg1 CILP 70:73 CILP 70:73 CILP 70:73 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 1 72 theme eukaryotic 190:199 arg1 cells 201:205 eukaryotic cells 190:205 eukaryotic cells 190:205 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 7 73 theme NTPPHase 991:998 arg1 homolog 972:978 The human homolog 962:978 The human homolog of porcine NTPPHase described here 962:1013 The human homolog of porcine NTPPHase described here contains 10 cysteine residues and two putative N-glycosylation sites. 9722584 3 74 theme kDa 475:477 arg1 mass 461:464 a calculated molecular mass 438:464 a calculated molecular mass of 132.5 kDa 438:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 0 75 theme novel 45:49 arg1 protein 61:67 a novel cartilage protein 43:67 a novel cartilage protein (CILP) 43:74 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 0 75 theme novel 45:49 arg1 CILP 70:73 CILP 70:73 CILP 70:73 Cloning and deduced amino acid sequence of a novel cartilage protein (CILP) identifies a proform including a nucleotide pyrophosphohydrolase. 9722584 9 76 theme full-length 1252:1262 arg1 cDNA 1264:1267 the full-length cDNA 1248:1267 the full-length cDNA 1248:1267 Expression of the full-length cDNA in a cell-free translation system and in COS-7 or EBNA cells indicates that the precursor protein is synthesized as a single polypeptide chain that is processed, possibly by a furin-like protease, into two polypeptides upon or preceding secretion. 9722584 3 77 theme 2-kilobase 337:346 arg1 mRNA 348:351 2-kilobase mRNA 337:351 2-kilobase mRNA detected in human articular cartilage 337:389 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 3 77 theme 2-kilobase 337:346 arg1 polypeptide 401:411 a polypeptide 399:411 a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa 399:477 2-kilobase mRNA detected in human articular cartilage encodes a polypeptide of 1184 amino acids with a calculated molecular mass of 132.5 kDa. 9722584 1 78 theme novel 212:216 arg1 protein 296:302 cartilage intermediate layer protein 267:302 cartilage intermediate layer protein (CILP) 267:309 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 9722584 1 78 theme novel 212:216 arg1 protein 218:224 a novel protein 210:224 a novel protein isolated from human articular cartilage 210:264 The cDNA cloning and expression in vitro and in eukaryotic cells of a novel protein isolated from human articular cartilage, cartilage intermediate layer protein (CILP) is described. 2243102 2 0 theme membrane 136:143 arg1 lamp-1 160:165 lamp-1 160:165 lamp-1 160:165 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 0 theme membrane 136:143 arg1 glycoproteins 145:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 0 theme membrane 136:143 arg1 lamp-2 171:176 lamp-2 171:176 lamp-2 171:176 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 1 mod modified 245:252 arg1 some 227:230 some 227:230 some 227:230 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 1 mod modified 245:252 arg1 N-glycans 216:224 18 and 16 N-glycans 206:224 18 and 16 N-glycans 206:224 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 1 mod modified 245:252 arg3 poly-N-acetyl-lactosamine 257:281 poly-N-acetyl-lactosamine 257:281 poly-N-acetyl-lactosamine 257:281 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 1 2 from Localization 88:99 arg1 peptide 108:114 the peptide 104:114 the peptide 104:114 Localization on the peptide backbones. 2243102 4 3 theme gel 620:622 arg1 filtration 624:633 gel filtration 620:633 gel filtration 620:633 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 3 4 theme specific 332:339 arg1 sites 341:345 specific sites 332:345 specific sites 332:345 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 2243102 2 5 theme Lysosome 127:134 arg1 lamp-1 160:165 lamp-1 160:165 lamp-1 160:165 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 5 theme Lysosome 127:134 arg1 glycoproteins 145:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 5 theme Lysosome 127:134 arg1 lamp-2 171:176 lamp-2 171:176 lamp-2 171:176 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 5 6 theme poly-N-acetyllactosaminyl 698:722 arg1 structures 724:733 The poly-N-acetyllactosaminyl structures 694:733 The poly-N-acetyllactosaminyl structures of isolated glycopeptides 694:759 The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. 2243102 4 7 theme glycoproteins 534:546 arg1 digestion 517:525 trypsin, pepsin, and V8 protease digestion 484:525 trypsin, pepsin, and V8 protease digestion of the glycoproteins 484:546 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 5 8 theme released 807:814 arg1 oligosaccharides 816:831 their released oligosaccharides 801:831 their released oligosaccharides 801:831 The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. 2243102 4 9 theme V8 505:506 arg1 digestion 517:525 trypsin, pepsin, and V8 protease digestion 484:525 trypsin, pepsin, and V8 protease digestion of the glycoproteins 484:546 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 7 10 theme glycosylation 1115:1127 arg1 sites 1129:1133 only certain glycosylation sites 1102:1133 only certain glycosylation sites 1102:1133 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 4 11 theme Polylactosaminoglycan-containing 424:455 arg1 glycopeptides 457:469 Polylactosaminoglycan-containing glycopeptides 424:469 Polylactosaminoglycan-containing glycopeptides 424:469 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 5 12 theme oligosaccharides 816:831 arg1 susceptibility 783:796 the susceptibility 779:796 the susceptibility of their released oligosaccharides to endo-beta-galactosidase 779:858 The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. 2243102 4 13 theme performance 659:669 arg1 chromatography 678:691 reverse phase high performance liquid chromatography 640:691 reverse phase high performance liquid chromatography 640:691 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 7 14 theme 1----3-N-acetylglucosaminyl 1243:1269 arg1 transferase 1271:1281 beta 1----3-N-acetylglucosaminyl transferase 1238:1281 beta 1----3-N-acetylglucosaminyl transferase 1238:1281 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 6 15 from Asn-34 953:958 arg1 lamp-1 1014:1019 lamp-1 1014:1019 lamp-1 1014:1019 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 6 15 from Asn-34 953:958 arg1 located 942:948 located 942:948 located 942:948 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 4 16 gly glycoproteins 534:546 arg1 glycoproteins 534:546 the glycoproteins 530:546 the glycoproteins 530:546 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 3 17 theme chronic 388:394 arg1 leukemia 408:415 chronic myelogenous leukemia 388:415 human chronic myelogenous leukemia cells 382:421 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 2243102 4 18 theme protease 508:515 arg1 digestion 517:525 trypsin, pepsin, and V8 protease digestion 484:525 trypsin, pepsin, and V8 protease digestion of the glycoproteins 484:546 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 6 19 from Asn-4 1029:1033 arg1 located 942:948 located 942:948 located 942:948 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 6 20 from located 942:948 arg1 lamp-2 1065:1070 lamp-2 1065:1070 lamp-2 1065:1070 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 3 21 theme myelogenous 396:406 arg1 leukemia 408:415 chronic myelogenous leukemia 388:415 human chronic myelogenous leukemia cells 382:421 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 2243102 4 22 theme agglutinin 584:593 arg1 chromatography 604:617 Datura stramonium agglutinin affinity chromatography 566:617 Datura stramonium agglutinin affinity chromatography 566:617 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 0 23 theme human 30:34 arg1 glycoproteins 55:67 human lysosomal membrane glycoproteins 30:67 human lysosomal membrane glycoproteins 30:67 The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. 2243102 4 24 theme stramonium 573:582 arg1 chromatography 604:617 Datura stramonium agglutinin affinity chromatography 566:617 Datura stramonium agglutinin affinity chromatography 566:617 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 6 25 from Asn-279 1054:1060 arg1 located 942:948 located 942:948 located 942:948 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 2 26 gly glycoproteins 145:157 arg1 lamp-1 160:165 lamp-1 160:165 lamp-1 160:165 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 26 gly glycoproteins 145:157 arg1 glycoproteins 145:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 26 gly glycoproteins 145:157 arg1 lamp-2 171:176 lamp-2 171:176 lamp-2 171:176 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 4 27 theme liquid 671:676 arg1 chromatography 678:691 reverse phase high performance liquid chromatography 640:691 reverse phase high performance liquid chromatography 640:691 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 3 28 theme human 382:386 arg1 cells 417:421 human chronic myelogenous leukemia cells 382:421 human chronic myelogenous leukemia cells 382:421 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 2243102 2 29 contain contain 198:204 arg1 lamp-1 160:165 lamp-1 160:165 lamp-1 160:165 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 29 contain contain 198:204 arg1 glycoproteins 145:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins 127:157 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 29 contain contain 198:204 arg2 N-glycans 216:224 18 and 16 N-glycans 206:224 18 and 16 N-glycans 206:224 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 2 29 contain contain 198:204 arg1 lamp-2 171:176 lamp-2 171:176 lamp-2 171:176 Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. 2243102 0 30 theme membrane 46:53 arg1 glycoproteins 55:67 human lysosomal membrane glycoproteins 30:67 human lysosomal membrane glycoproteins 30:67 The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. 2243102 4 31 theme affinity 595:602 arg1 chromatography 604:617 Datura stramonium agglutinin affinity chromatography 566:617 Datura stramonium agglutinin affinity chromatography 566:617 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 6 32 from lamp-2 1065:1070 arg1 located 942:948 located 942:948 located 942:948 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 4 33 theme trypsin 484:490 arg1 digestion 517:525 trypsin, pepsin, and V8 protease digestion 484:525 trypsin, pepsin, and V8 protease digestion of the glycoproteins 484:546 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 0 34 dep lamp-1 69:74 arg1 polylactosaminoglycans 4:25 The polylactosaminoglycans 0:25 The polylactosaminoglycans of human lysosomal membrane glycoproteins 0:67 The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. 2243102 0 35 theme lysosomal 36:44 arg1 glycoproteins 55:67 human lysosomal membrane glycoproteins 30:67 human lysosomal membrane glycoproteins 30:67 The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. 2243102 7 36 gly glycosylation 1115:1127 arg2 sites 1129:1133 only certain glycosylation sites 1102:1133 only certain glycosylation sites 1102:1133 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 6 37 theme acid 867:870 arg1 analysis 872:879 Amino acid analysis 861:879 Amino acid analysis 861:879 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 3 38 theme leukemia 408:415 arg1 cells 417:421 human chronic myelogenous leukemia cells 382:421 human chronic myelogenous leukemia cells 382:421 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 2243102 5 39 theme isolated 738:745 arg1 glycopeptides 747:759 isolated glycopeptides 738:759 isolated glycopeptides 738:759 The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. 2243102 4 40 theme pepsin 493:498 arg1 digestion 517:525 trypsin, pepsin, and V8 protease digestion 484:525 trypsin, pepsin, and V8 protease digestion of the glycoproteins 484:546 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 6 41 theme Amino 861:865 arg1 analysis 872:879 Amino acid analysis 861:879 Amino acid analysis 861:879 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 0 42 theme glycoproteins 55:67 arg1 polylactosaminoglycans 4:25 The polylactosaminoglycans 0:25 The polylactosaminoglycans of human lysosomal membrane glycoproteins 0:67 The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. 2243102 4 43 theme Datura 566:571 arg1 chromatography 604:617 Datura stramonium agglutinin affinity chromatography 566:617 Datura stramonium agglutinin affinity chromatography 566:617 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 5 44 theme glycopeptides 747:759 arg1 structures 724:733 The poly-N-acetyllactosaminyl structures 694:733 The poly-N-acetyllactosaminyl structures of isolated glycopeptides 694:759 The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. 2243102 7 45 mod modified 1154:1161 arg1 sites 1129:1133 only certain glycosylation sites 1102:1133 only certain glycosylation sites 1102:1133 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 7 45 mod modified 1154:1161 arg3 poly-N-acetyllactosamine 1166:1189 poly-N-acetyllactosamine 1166:1189 poly-N-acetyllactosamine 1166:1189 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 4 46 theme reverse 640:646 arg1 chromatography 678:691 reverse phase high performance liquid chromatography 640:691 reverse phase high performance liquid chromatography 640:691 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 6 47 from Asn-10 1042:1047 arg1 located 942:948 located 942:948 located 942:948 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 7 48 theme certain 1107:1113 arg1 sites 1129:1133 only certain glycosylation sites 1102:1133 only certain glycosylation sites 1102:1133 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 6 49 located located 942:948 arg2 polylactosaminoglycans 914:935 polylactosaminoglycans 914:935 polylactosaminoglycans 914:935 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 6 49 located located 942:948 arg1 Asn-34 953:958 Asn-34 953:958 Asn-34 953:958 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 6 49 located located 942:948 arg1 lamp-2 1065:1070 lamp-2 1065:1070 lamp-2 1065:1070 Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. 2243102 0 50 gly glycoproteins 55:67 arg1 glycoproteins 55:67 human lysosomal membrane glycoproteins 30:67 human lysosomal membrane glycoproteins 30:67 The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. 2243102 5 51 gly glycopeptides 747:759 arg2 glycopeptides 747:759 isolated glycopeptides 738:759 isolated glycopeptides 738:759 The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. 2243102 4 52 theme high 654:657 arg1 chromatography 678:691 reverse phase high performance liquid chromatography 640:691 reverse phase high performance liquid chromatography 640:691 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 7 53 theme beta 1238:1241 arg1 transferase 1271:1281 beta 1----3-N-acetylglucosaminyl transferase 1238:1281 beta 1----3-N-acetylglucosaminyl transferase 1238:1281 These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase. 2243102 4 54 theme phase 648:652 arg1 chromatography 678:691 reverse phase high performance liquid chromatography 640:691 reverse phase high performance liquid chromatography 640:691 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 4 55 gly glycopeptides 457:469 arg2 glycopeptides 457:469 Polylactosaminoglycan-containing glycopeptides 424:469 Polylactosaminoglycan-containing glycopeptides 424:469 Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. 2243102 3 56 from polylactosaminoglycans 306:327 arg1 lamp-2 361:366 lamp-2 361:366 lamp-2 361:366 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 2243102 3 56 from polylactosaminoglycans 306:327 arg1 lamp-1 350:355 lamp-1 350:355 lamp-1 350:355 We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. 1533657 6 0 theme H-related 851:859 arg1 h42 878:880 h42 878:880 h42 878:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 0 theme H-related 851:859 arg1 h37 870:872 h37 870:872 h37 870:872 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 0 theme H-related 851:859 arg1 proteins 861:868 the previously described factor H-related proteins h37 and h42 819:880 the previously described factor H-related proteins h37 and h42 819:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 7 1 theme acids 1045:1049 arg1 sequences 1007:1015 identical leader sequences 990:1015 identical leader sequences 990:1015 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 1 theme acids 1045:1049 arg1 degree 1101:1106 a lower degree 1093:1106 a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42) 1093:1172 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 1 theme acids 1045:1049 arg1 sequences 1070:1078 identical sequences 1060:1078 identical sequences of SCR2 1060:1086 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 1 theme acids 1045:1049 arg1 exchange 1021:1028 an exchange 1018:1028 an exchange of three amino acids in SCR1 1018:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 2 2 theme glycosylated 327:338 arg1 forms 340:344 differently glycosylated forms 315:344 differently glycosylated forms 315:344 These proteins represent differently glycosylated forms and are encoded by the same mRNA. 1533657 1 3 theme kDa 229:231 arg1 mass 211:214 an apparent molecular mass 189:214 an apparent molecular mass of 24 and 29 kDa 189:231 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 6 4 theme described 834:842 arg1 h42 878:880 h42 878:880 h42 878:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 4 theme described 834:842 arg1 h37 870:872 h37 870:872 h37 870:872 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 4 theme described 834:842 arg1 proteins 861:868 the previously described factor H-related proteins h37 and h42 819:880 the previously described factor H-related proteins h37 and h42 819:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 4 5 theme amino 548:552 arg1 acids 554:558 270 amino acids 544:558 270 amino acids 544:558 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 9 6 theme family 1464:1469 arg1 existence 1444:1452 the existence 1440:1452 the existence of a gene family consisting of factor H and at least two factor H-related genes 1440:1532 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 4 7 theme indicative 607:616 arg1 sequence 597:604 a hydrophobic leader sequence 576:604 a hydrophobic leader sequence 576:604 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 1 8 theme human 163:167 arg1 proteins 175:182 two human serum proteins 159:182 two human serum proteins with an apparent molecular mass of 24 and 29 kDa 159:231 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 7 9 theme SCR2 1083:1086 arg1 sequences 1007:1015 identical leader sequences 990:1015 identical leader sequences 990:1015 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 9 theme SCR2 1083:1086 arg1 degree 1101:1106 a lower degree 1093:1106 a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42) 1093:1172 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 9 theme SCR2 1083:1086 arg1 sequences 1070:1078 identical sequences 1060:1078 identical sequences of SCR2 1060:1086 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 9 theme SCR2 1083:1086 arg1 exchange 1021:1028 an exchange 1018:1028 an exchange of three amino acids in SCR1 1018:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 9 10 theme elements 1299:1306 arg1 relatedness 1273:1283 The relatedness 1269:1283 The relatedness 1269:1283 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 4 11 theme hydrophobic 578:588 arg1 sequence 597:604 a hydrophobic leader sequence 576:604 a hydrophobic leader sequence 576:604 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 8 12 theme h29 1206:1208 arg1 SCR3-4 1188:1193 SCR3-4 1188:1193 SCR3-4 of h24 and h29 1188:1208 In addition, SCR3-4 of h24 and h29 display homology to SCR19-20 of human complement factor H. 1533657 7 13 theme leader 1000:1005 arg1 sequences 1007:1015 identical leader sequences 990:1015 identical leader sequences 990:1015 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 0 14 theme genes 123:127 arg1 family 96:101 a family 94:101 a family of factor H-related genes 94:127 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 5 15 theme glycosylation 745:757 arg1 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 7 16 theme acid 932:935 arg1 comparison 937:946 Amino acid comparison 926:946 Amino acid comparison of these factor H-related proteins 926:981 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 1 17 theme molecular 201:209 arg1 mass 211:214 an apparent molecular mass 189:214 an apparent molecular mass of 24 and 29 kDa 189:231 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 7 18 from degree 1101:1106 arg1 SCR1 1054:1057 SCR1 1054:1057 SCR1 1054:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 19 theme H-related 964:972 arg1 proteins 974:981 these factor H-related proteins 951:981 these factor H-related proteins 951:981 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 9 20 theme H-related 1518:1526 arg1 genes 1528:1532 at least two factor H-related genes 1498:1532 at least two factor H-related genes 1498:1532 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 6 21 attach derived 898:904 arg2 proteins 861:868 the previously described factor H-related proteins h37 and h42 819:880 the previously described factor H-related proteins h37 and h42 819:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 21 attach derived 898:904 arg2 h37 870:872 h37 870:872 h37 870:872 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 21 attach derived 898:904 arg1 mRNA 920:923 a 1.4-kb mRNA 911:923 a 1.4-kb mRNA 911:923 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 21 attach derived 898:904 arg2 h42 878:880 h42 878:880 h42 878:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 4 22 theme protein 632:638 arg1 indicative 607:616 indicative 607:616 indicative 607:616 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 6 23 dep proteins 861:868 arg1 h42 878:880 h42 878:880 h42 878:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 23 dep proteins 861:868 arg1 h37 870:872 h37 870:872 h37 870:872 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 23 dep proteins 861:868 arg1 proteins 861:868 the previously described factor H-related proteins h37 and h42 819:880 the previously described factor H-related proteins h37 and h42 819:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 5 24 link N-linked 736:743 arg1 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 7 25 from SCR1 1054:1057 arg1 sequences 1007:1015 identical leader sequences 990:1015 identical leader sequences 990:1015 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 25 from SCR1 1054:1057 arg1 degree 1101:1106 a lower degree 1093:1106 a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42) 1093:1172 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 25 from SCR1 1054:1057 arg1 sequences 1070:1078 identical sequences 1060:1078 identical sequences of SCR2 1060:1086 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 25 from SCR1 1054:1057 arg1 exchange 1021:1028 an exchange 1018:1028 an exchange of three amino acids in SCR1 1018:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 0 26 theme H. 78:79 arg1 Evidence 81:88 factor H. Evidence 71:88 factor H. Evidence for a family of factor H-related genes 71:127 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 7 27 theme identical 990:998 arg1 sequences 1007:1015 identical leader sequences 990:1015 identical leader sequences 990:1015 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 6 28 theme 1.4-kb 913:918 arg1 mRNA 920:923 a 1.4-kb mRNA 911:923 a 1.4-kb mRNA 911:923 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 2 29 theme same 369:372 arg1 mRNA 374:377 the same mRNA 365:377 the same mRNA 365:377 These proteins represent differently glycosylated forms and are encoded by the same mRNA. 1533657 5 30 theme consensus 686:694 arg1 SCR 705:707 SCR 705:707 SCR 705:707 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 5 30 theme consensus 686:694 arg1 repeats 696:702 four short consensus repeats 675:702 four short consensus repeats (SCR) 675:708 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 9 31 theme factor 1371:1376 arg1 H 1378:1378 factor H 1371:1378 factor H 1371:1378 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 3 32 theme cDNA 398:401 arg1 clone 403:407 The corresponding cDNA clone 380:407 The corresponding cDNA clone 380:407 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 3 32 theme cDNA 398:401 arg1 bp 417:418 1051 bp 412:418 1051 bp in size 412:426 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 5 33 theme N-linked 736:743 arg1 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 4 34 theme translation 500:510 arg1 product 512:518 The predicted translation product 486:518 The predicted translation product 486:518 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 0 35 theme human 15:19 arg1 proteins 27:34 Two additional human serum proteins 0:34 Two additional human serum proteins structurally related 0:55 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 7 36 from exchange 1021:1028 arg1 SCR1 1054:1057 SCR1 1054:1057 SCR1 1054:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 2 37 gly glycosylated 327:338 arg1 forms 340:344 differently glycosylated forms 315:344 differently glycosylated forms 315:344 These proteins represent differently glycosylated forms and are encoded by the same mRNA. 1533657 7 38 theme identical 1060:1068 arg1 sequences 1070:1078 identical sequences 1060:1078 identical sequences of SCR2 1060:1086 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 1 39 theme complement 269:278 arg1 H 287:287 complement factor H 269:287 complement factor H 269:287 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 9 40 theme common 1401:1406 arg1 function 1392:1399 a function 1390:1399 a function common to these proteins 1390:1424 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 8 41 theme complement 1248:1257 arg1 H 1266:1266 human complement factor H 1242:1266 human complement factor H 1242:1266 In addition, SCR3-4 of h24 and h29 display homology to SCR19-20 of human complement factor H. 1533657 5 42 theme single 720:725 arg1 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 1 43 with proteins 175:182 arg1 mass 211:214 an apparent molecular mass 189:214 an apparent molecular mass of 24 and 29 kDa 189:231 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 3 44 attach derived 460:466 arg1 liver 479:483 human liver 473:483 human liver 473:483 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 3 44 attach derived 460:466 arg2 mRNA 455:458 a 1.4-kb mRNA 446:458 a 1.4-kb mRNA derived from human liver 446:483 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 7 45 from acids 1045:1049 arg1 SCR1 1054:1057 SCR1 1054:1057 SCR1 1054:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 8 46 theme H 1266:1266 arg1 SCR19-20 1230:1237 SCR19-20 1230:1237 SCR19-20 of human complement factor H 1230:1266 In addition, SCR3-4 of h24 and h29 display homology to SCR19-20 of human complement factor H. 1533657 4 47 theme acids 554:558 arg1 protein 533:539 a protein 531:539 a protein 531:539 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 7 48 theme amino 1039:1043 arg1 acids 1045:1049 three amino acids 1033:1049 three amino acids in SCR1 1033:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 6 49 theme factor 844:849 arg1 h42 878:880 h42 878:880 h42 878:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 49 theme factor 844:849 arg1 h37 870:872 h37 870:872 h37 870:872 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 6 49 theme factor 844:849 arg1 proteins 861:868 the previously described factor H-related proteins h37 and h42 819:880 the previously described factor H-related proteins h37 and h42 819:880 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 9 50 theme gene 1459:1462 arg1 family 1464:1469 a gene family 1457:1469 a gene family consisting of factor H and at least two factor H-related genes 1457:1532 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 9 51 theme structural 1288:1297 arg1 elements 1299:1306 structural elements 1288:1306 structural elements of the factor H-related proteins h24, h29, h37, and h42 1288:1362 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 0 52 theme related 49:55 arg1 proteins 27:34 Two additional human serum proteins 0:34 Two additional human serum proteins structurally related 0:55 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 7 53 theme homology 1111:1118 arg1 sequences 1007:1015 identical leader sequences 990:1015 identical leader sequences 990:1015 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 53 theme homology 1111:1118 arg1 degree 1101:1106 a lower degree 1093:1106 a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42) 1093:1172 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 53 theme homology 1111:1118 arg1 sequences 1070:1078 identical sequences 1060:1078 identical sequences of SCR2 1060:1086 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 53 theme homology 1111:1118 arg1 exchange 1021:1028 an exchange 1018:1028 an exchange of three amino acids in SCR1 1018:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 1 54 theme serum 169:173 arg1 proteins 175:182 two human serum proteins 159:182 two human serum proteins with an apparent molecular mass of 24 and 29 kDa 159:231 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 9 55 theme factor 1485:1490 arg1 H 1492:1492 factor H 1485:1492 factor H 1485:1492 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 5 56 theme secreted 645:652 arg1 part 654:657 The secreted part 641:657 The secreted part 641:657 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 5 57 gly glycosylation 745:757 arg2 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 7 58 theme proteins 974:981 arg1 comparison 937:946 Amino acid comparison 926:946 Amino acid comparison of these factor H-related proteins 926:981 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 9 59 theme factor 1315:1320 arg1 proteins 1332:1339 the factor H-related proteins 1311:1339 the factor H-related proteins h24, h29, h37, and h42 1311:1362 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 4 60 theme leader 590:595 arg1 sequence 597:604 a hydrophobic leader sequence 576:604 a hydrophobic leader sequence 576:604 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 7 61 theme factor 957:962 arg1 proteins 974:981 these factor H-related proteins 951:981 these factor H-related proteins 951:981 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 9 62 theme proteins 1332:1339 arg1 elements 1299:1306 structural elements 1288:1306 structural elements of the factor H-related proteins h24, h29, h37, and h42 1288:1362 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 0 63 theme H-related 113:121 arg1 genes 123:127 factor H-related genes 106:127 factor H-related genes 106:127 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 9 64 theme factor 1511:1516 arg1 genes 1528:1532 at least two factor H-related genes 1498:1532 at least two factor H-related genes 1498:1532 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 7 65 theme Amino 926:930 arg1 comparison 937:946 Amino acid comparison 926:946 Amino acid comparison of these factor H-related proteins 926:981 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 5 66 contain has 714:716 arg2 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 5 66 contain has 714:716 arg1 part 654:657 The secreted part 641:657 The secreted part 641:657 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 0 67 theme factor 71:76 arg1 Evidence 81:88 factor H. Evidence 71:88 factor H. Evidence for a family of factor H-related genes 71:127 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 4 68 theme secreted 623:630 arg1 protein 632:638 a secreted protein 621:638 a secreted protein 621:638 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 0 69 theme factor 106:111 arg1 genes 123:127 factor H-related genes 106:127 factor H-related genes 106:127 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 7 70 theme lower 1095:1099 arg1 degree 1101:1106 a lower degree 1093:1106 a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42) 1093:1172 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 71 from sequences 1070:1078 arg1 SCR1 1054:1057 SCR1 1054:1057 SCR1 1054:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 5 72 theme short 680:684 arg1 SCR 705:707 SCR 705:707 SCR 705:707 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 5 72 theme short 680:684 arg1 repeats 696:702 four short consensus repeats 675:702 four short consensus repeats (SCR) 675:708 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 7 73 dep SCR4-5 1153:1158 arg1 h42 1169:1171 h42 1169:1171 h42 1169:1171 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 73 dep SCR4-5 1153:1158 arg1 h37 1161:1163 h37 1161:1163 h37 1161:1163 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 8 74 theme h24 1198:1200 arg1 SCR3-4 1188:1193 SCR3-4 1188:1193 SCR3-4 of h24 and h29 1188:1208 In addition, SCR3-4 of h24 and h29 display homology to SCR19-20 of human complement factor H. 1533657 5 75 theme putative 727:734 arg1 site 759:762 a single putative N-linked glycosylation site 718:762 a single putative N-linked glycosylation site 718:762 The secreted part is organized in four short consensus repeats (SCR) and has a single putative N-linked glycosylation site. 1533657 9 76 theme H 1378:1378 arg1 relatedness 1273:1283 The relatedness 1269:1283 The relatedness 1269:1283 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 7 77 from sequences 1007:1015 arg1 SCR1 1054:1057 SCR1 1054:1057 SCR1 1054:1057 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 1 78 dep characterize 146:157 arg1 related 258:264 related 258:264 related 258:264 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 0 79 theme serum 21:25 arg1 proteins 27:34 Two additional human serum proteins 0:34 Two additional human serum proteins structurally related 0:55 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 1533657 1 80 theme apparent 192:199 arg1 mass 211:214 an apparent molecular mass 189:214 an apparent molecular mass of 24 and 29 kDa 189:231 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 3 81 theme corresponding 384:396 arg1 clone 403:407 The corresponding cDNA clone 380:407 The corresponding cDNA clone 380:407 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 3 81 theme corresponding 384:396 arg1 bp 417:418 1051 bp 412:418 1051 bp in size 412:426 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 7 82 dep SCR3-4 1128:1133 arg1 h29 1144:1146 h29 1144:1146 h29 1144:1146 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 7 82 dep SCR3-4 1128:1133 arg1 h24 1136:1138 h24 1136:1138 h24 1136:1138 Amino acid comparison of these factor H-related proteins showed identical leader sequences, an exchange of three amino acids in SCR1, identical sequences of SCR2, and a lower degree of homology between SCR3-4 (h24 and h29) and SCR4-5 (h37 and h42). 1533657 3 83 from bp 417:418 arg1 size 423:426 size 423:426 size 423:426 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 1 84 theme factor 280:285 arg1 H 287:287 complement factor H 269:287 complement factor H 269:287 We identify and characterize two human serum proteins with an apparent molecular mass of 24 and 29 kDa, which are antigenically related to complement factor H. 1533657 4 85 theme predicted 490:498 arg1 product 512:518 The predicted translation product 486:518 The predicted translation product 486:518 The predicted translation product represents a protein of 270 amino acids, which displays a hydrophobic leader sequence, indicative of a secreted protein. 1533657 8 86 theme human 1242:1246 arg1 H 1266:1266 human complement factor H 1242:1266 human complement factor H 1242:1266 In addition, SCR3-4 of h24 and h29 display homology to SCR19-20 of human complement factor H. 1533657 3 87 theme 1.4-kb 448:453 arg1 mRNA 455:458 a 1.4-kb mRNA 446:458 a 1.4-kb mRNA derived from human liver 446:483 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 6 88 theme predicted 770:778 arg1 sequence 780:787 This predicted sequence 765:787 This predicted sequence 765:787 This predicted sequence is closely related to that of the previously described factor H-related proteins h37 and h42, which are also derived from a 1.4-kb mRNA. 1533657 8 89 theme factor 1259:1264 arg1 H 1266:1266 human complement factor H 1242:1266 human complement factor H 1242:1266 In addition, SCR3-4 of h24 and h29 display homology to SCR19-20 of human complement factor H. 1533657 3 90 theme human 473:477 arg1 liver 479:483 human liver 473:483 human liver 473:483 The corresponding cDNA clone is 1051 bp in size and hybridized to a 1.4-kb mRNA derived from human liver. 1533657 9 91 theme H-related 1322:1330 arg1 proteins 1332:1339 the factor H-related proteins 1311:1339 the factor H-related proteins h24, h29, h37, and h42 1311:1362 The relatedness of structural elements of the factor H-related proteins h24, h29, h37, and h42 and of factor H, suggests a function common to these proteins and indicates the existence of a gene family consisting of factor H and at least two factor H-related genes. 1533657 0 92 theme additional 4:13 arg1 proteins 27:34 Two additional human serum proteins 0:34 Two additional human serum proteins structurally related 0:55 Two additional human serum proteins structurally related to complement factor H. Evidence for a family of factor H-related genes. 6333684 4 0 theme mapping 711:717 arg1 combination 688:698 a combination 686:698 a combination of peptide mapping and protein sequencing 686:740 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 1 1 theme immunoaffinity 157:170 arg1 chromatography 172:185 selective immunoaffinity chromatography 147:185 selective immunoaffinity chromatography 147:185 Human interleukin 2 was separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. 6333684 3 2 theme significant 522:532 arg1 effect 534:539 no significant effect 519:539 no significant effect 519:539 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 4 3 theme interleukin 590:600 arg1 sequence 578:585 The entire primary sequence 559:585 The entire primary sequence 559:585 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 3 4 theme molecular 370:378 arg1 forms 380:384 The various molecular forms 358:384 The various molecular forms of interleukin 2 358:401 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 2 5 theme residue 310:316 arg1 glycosylation 279:291 glycosylation 279:291 glycosylation of the threonine residue in position 3 of the polypeptide chain 279:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 2 6 theme threonine 300:308 arg1 residue 310:316 the threonine residue 296:316 the threonine residue in position 3 of the polypeptide chain 296:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 5 7 theme s 811:811 arg1 determination 778:790 the determination 774:790 the determination of the active site(s) of the molecule 774:828 This information should aid in the determination of the active site(s) of the molecule. 6333684 3 8 theme in 451:452 arg1 assay 474:478 the in vitro proliferation assay 447:478 the in vitro proliferation assay 447:478 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 3 9 theme various 362:368 arg1 forms 380:384 The various molecular forms 358:384 The various molecular forms of interleukin 2 358:401 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 3 10 contain had 515:517 arg1 glycosylation 501:513 the glycosylation 497:513 the glycosylation 497:513 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 3 10 contain had 515:517 arg2 effect 534:539 no significant effect 519:539 no significant effect 519:539 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 1 11 theme Human 80:84 arg1 interleukin 86:96 Human interleukin 2 80:98 Human interleukin 2 80:98 Human interleukin 2 was separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. 6333684 0 12 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and post-translational modification of human interleukin 2. 6333684 0 13 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and post-translational modification of human interleukin 2. 6333684 4 14 theme primary 570:576 arg1 sequence 578:585 The entire primary sequence 559:585 The entire primary sequence 559:585 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 3 15 contain had 403:405 arg2 activities 433:442 nearly identical specific activities 407:442 nearly identical specific activities 407:442 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 3 15 contain had 403:405 arg1 forms 380:384 The various molecular forms 358:384 The various molecular forms of interleukin 2 358:401 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 2 16 gly glycosylation 279:291 arg1 residue 310:316 the threonine residue 296:316 the threonine residue in position 3 of the polypeptide chain 296:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 2 17 from variations 265:274 arg1 glycosylation 279:291 glycosylation 279:291 glycosylation of the threonine residue in position 3 of the polypeptide chain 279:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 4 18 theme entire 563:568 arg1 sequence 578:585 The entire primary sequence 559:585 The entire primary sequence 559:585 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 5 19 theme site 806:809 arg1 s 811:811 the active site(s) 795:812 the active site(s) of the molecule 795:828 This information should aid in the determination of the active site(s) of the molecule. 6333684 4 20 theme disulfide 650:658 arg1 bridge 660:665 the intramolecular disulfide bridge 631:665 the intramolecular disulfide bridge 631:665 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 3 21 theme interleukin 389:399 arg1 forms 380:384 The various molecular forms 358:384 The various molecular forms of interleukin 2 358:401 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 4 22 theme peptide 703:709 arg1 mapping 711:717 peptide mapping 703:717 peptide mapping 703:717 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 2 23 from glycosylation 279:291 arg1 position 321:328 position 3 321:330 position 3 of the polypeptide chain 321:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 4 24 theme intramolecular 635:648 arg1 bridge 660:665 the intramolecular disulfide bridge 631:665 the intramolecular disulfide bridge 631:665 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 3 25 theme specific 424:431 arg1 activities 433:442 nearly identical specific activities 407:442 nearly identical specific activities 407:442 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 2 26 theme chain 351:355 arg1 position 321:328 position 3 321:330 position 3 of the polypeptide chain 321:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 3 27 theme proliferation 460:472 arg1 assay 474:478 the in vitro proliferation assay 447:478 the in vitro proliferation assay 447:478 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 0 28 theme post-translational 24:41 arg1 modification 43:54 post-translational modification 24:54 post-translational modification of human interleukin 2 24:77 Amino acid sequence and post-translational modification of human interleukin 2. 6333684 5 29 theme active 799:804 arg1 s 811:811 the active site(s) 795:812 the active site(s) of the molecule 795:828 This information should aid in the determination of the active site(s) of the molecule. 6333684 4 30 theme bridge 660:665 arg1 location 619:626 the location 615:626 the location of the intramolecular disulfide bridge 615:665 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 0 31 theme human 59:63 arg1 interleukin 65:75 human interleukin 2 59:77 human interleukin 2 59:77 Amino acid sequence and post-translational modification of human interleukin 2. 6333684 3 32 dep in 451:452 arg1 vitro 454:458 vitro 454:458 vitro 454:458 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 4 33 theme protein 723:729 arg1 sequencing 731:740 protein sequencing 723:740 protein sequencing 723:740 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 3 34 theme identical 414:422 arg1 activities 433:442 nearly identical specific activities 407:442 nearly identical specific activities 407:442 The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. 6333684 1 35 theme multiple 119:126 arg1 forms 138:142 multiple molecular forms 119:142 multiple molecular forms 119:142 Human interleukin 2 was separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. 6333684 1 36 theme molecular 128:136 arg1 forms 138:142 multiple molecular forms 119:142 multiple molecular forms 119:142 Human interleukin 2 was separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. 6333684 2 37 theme most 217:220 arg1 part 222:225 the most part 213:225 the most part 213:225 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 4 38 theme sequencing 731:740 arg1 combination 688:698 a combination 686:698 a combination of peptide mapping and protein sequencing 686:740 The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. 6333684 2 39 from residue 310:316 arg1 position 321:328 position 3 321:330 position 3 of the polypeptide chain 321:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 2 40 theme polypeptide 339:349 arg1 chain 351:355 the polypeptide chain 335:355 the polypeptide chain 335:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 6333684 5 41 theme molecule 821:828 arg1 s 811:811 the active site(s) 795:812 the active site(s) of the molecule 795:828 This information should aid in the determination of the active site(s) of the molecule. 6333684 0 42 theme interleukin 65:75 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and post-translational modification of human interleukin 2. 6333684 0 42 theme interleukin 65:75 arg1 modification 43:54 post-translational modification 24:54 post-translational modification of human interleukin 2 24:77 Amino acid sequence and post-translational modification of human interleukin 2. 6333684 1 43 theme selective 147:155 arg1 chromatography 172:185 selective immunoaffinity chromatography 147:185 selective immunoaffinity chromatography 147:185 Human interleukin 2 was separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. 6333684 2 44 from position 321:328 arg1 glycosylation 279:291 glycosylation 279:291 glycosylation of the threonine residue in position 3 of the polypeptide chain 279:355 For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. 7607222 10 0 theme oligosaccharides 1363:1378 arg1 ends 1350:1353 The non-reducing ends 1333:1353 The non-reducing ends of some oligosaccharides 1333:1378 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 10 1 dep linked 1445:1450 arg1 alpha 1461:1465 alpha 1-2 1461:1469 alpha 1-2 1461:1469 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 10 1 dep linked 1445:1450 arg1 Fuc 1441:1443 external Fuc 1432:1443 external Fuc linked only in (alpha 1-2) to Gal 1432:1477 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 2 2 theme pathological 354:365 arg1 conditions 367:376 pathological conditions 354:376 pathological conditions 354:376 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 2 2 theme pathological 354:365 arg1 disease 399:405 Alzheimer's disease 387:405 Alzheimer's disease (brain) 387:413 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 9 3 theme beta 1264:1267 arg1 GlcNAc 1257:1262 Gal(beta 1-4)GlcNAc 1244:1262 Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)] 1244:1287 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 9 3 theme beta 1264:1267 arg1 1-6 1269:1271 beta 1-6 1264:1271 beta 1-6 1264:1271 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 7 4 theme same 1058:1061 arg1 structure 1070:1078 the same core 2 structure 1054:1078 the same core 2 structure 1054:1078 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 13 5 theme pancreatic 2009:2018 arg1 ducts 2020:2024 the pancreatic ducts 2005:2024 the pancreatic ducts 2005:2024 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 2 6 theme stone 166:170 arg1 protein 172:178 pancreatic stone protein 155:178 pancreatic stone protein 155:178 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 2 7 theme exocrine 281:288 arg1 pancreas 290:297 the exocrine pancreas 277:297 the exocrine pancreas 277:297 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 4 8 theme glycan 618:623 arg1 moiety 625:630 its glycan moiety 614:630 its glycan moiety 614:630 Little is known about its glycan moiety, which conceivably might be involved in this tissue specificity and pathophysiological characteristics. 7607222 10 9 theme external 1432:1439 arg1 alpha 1461:1465 alpha 1-2 1461:1469 alpha 1-2 1461:1469 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 10 9 theme external 1432:1439 arg1 Fuc 1441:1443 external Fuc 1432:1443 external Fuc linked only in (alpha 1-2) to Gal 1432:1477 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 12 10 theme SDS 1725:1727 arg1 electrophoresis 1733:1747 SDS gel electrophoresis 1725:1747 SDS gel electrophoresis 1725:1747 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 2 11 dep glycoprotein 237:248 arg1 whereas 300:306 whereas 300:306 whereas 300:306 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 10 12 dep determinant 1400:1410 arg1 H 1412:1412 H 1412:1412 the antigenic determinant H 1386:1412 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 5 13 theme lithostathine 811:823 arg1 chains 784:789 the major oligosaccharide chains 758:789 the major oligosaccharide chains of human pancreatic lithostathine 758:823 Therefore we isolated the major oligosaccharide chains of human pancreatic lithostathine and determined their sequences by means of NMR analysis. 7607222 12 14 theme protein 1698:1704 arg1 polymorphism 1678:1689 the polymorphism 1674:1689 the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing 1674:1772 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 3 15 theme growth 541:546 arg1 factor 548:553 a growth factor 539:553 a growth factor which stimulates islet regeneration 539:589 In the latter case, it has been shown recently that it acts as a growth factor which stimulates islet regeneration. 7607222 3 15 theme growth 541:546 arg1 it 528:529 it 528:529 it 528:529 In the latter case, it has been shown recently that it acts as a growth factor which stimulates islet regeneration. 7607222 11 16 from acid 1524:1527 arg1 linkage 1544:1550 (alpha 2-3) linkage 1532:1550 (alpha 2-3) linkage to Gal 1532:1557 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 13 17 dep in 1910:1911 arg1 situ 1913:1916 situ 1913:1916 situ 1913:1916 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 12 18 theme enzymic 1810:1816 arg1 processing 1818:1827 enzymic processing 1810:1827 enzymic processing 1810:1827 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 2 19 gly glycoprotein 237:248 arg1 glycoprotein 237:248 a glycoprotein 235:248 a glycoprotein which is normally found in the exocrine pancreas 235:297 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 2 19 gly glycoprotein 237:248 arg1 Lithostathine 126:138 Lithostathine 126:138 Lithostathine 126:138 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 11 20 theme disialylated 1569:1580 arg1 form 1582:1585 one disialylated form 1565:1585 one disialylated form which displays a supplementary (alpha 2-6) linkage 1565:1636 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 7 21 theme same 1006:1009 arg1 glycosylation 1019:1031 glycosylation 1019:1031 glycosylation (Thr5) 1019:1038 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 7 21 theme same 1006:1009 arg1 site 1011:1014 the same site 1002:1014 the same site of glycosylation (Thr5) 1002:1038 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 11 22 contain carry 1509:1513 arg2 acid 1524:1527 a sialic acid 1515:1527 a sialic acid in (alpha 2-3) linkage to Gal 1515:1557 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 11 22 contain carry 1509:1513 arg1 glycans 1488:1494 All the glycans 1480:1494 All the glycans 1480:1494 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 13 23 dep bond 1900:1903 arg1 hydrolysis 1918:1927 in situ hydrolysis 1910:1927 in situ hydrolysis 1910:1927 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 3 24 theme islet 572:576 arg1 regeneration 578:589 islet regeneration 572:589 islet regeneration 572:589 In the latter case, it has been shown recently that it acts as a growth factor which stimulates islet regeneration. 7607222 12 25 theme native 1789:1794 arg1 form 1796:1799 its native form 1785:1799 its native form 1785:1799 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 5 26 theme NMR 868:870 arg1 analysis 872:879 NMR analysis 868:879 NMR analysis 868:879 Therefore we isolated the major oligosaccharide chains of human pancreatic lithostathine and determined their sequences by means of NMR analysis. 7607222 6 27 theme them 977:980 arg1 seven 968:972 seven 968:972 seven 968:972 We obtained eleven different glycoforms and we were able to determine the sequence of seven of them. 7607222 6 27 theme them 977:980 arg1 them 977:980 them 977:980 them 977:980 We obtained eleven different glycoforms and we were able to determine the sequence of seven of them. 7607222 7 28 theme GalNAc 1112:1117 arg1 alpha- 1119:1124 GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha- 1081:1124 GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha- 1081:1124 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 6 29 theme seven 968:972 arg1 sequence 956:963 the sequence 952:963 the sequence of seven of them 952:980 We obtained eleven different glycoforms and we were able to determine the sequence of seven of them. 7607222 11 30 theme 2-3 1539:1541 arg1 linkage 1544:1550 (alpha 2-3) linkage 1532:1550 (alpha 2-3) linkage to Gal 1532:1557 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 10 31 contain carry 1380:1384 arg1 ends 1350:1353 The non-reducing ends 1333:1353 The non-reducing ends of some oligosaccharides 1333:1378 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 10 31 contain carry 1380:1384 arg2 determinant 1400:1410 the antigenic determinant 1386:1410 the antigenic determinant H 1386:1412 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 1 32 theme possible 84:91 arg1 implications 112:123 possible pathophysiological implications 84:123 possible pathophysiological implications 84:123 Structure characterization and possible pathophysiological implications. 7607222 4 33 theme tissue 677:682 arg1 specificity 684:694 this tissue specificity 672:694 this tissue specificity 672:694 Little is known about its glycan moiety, which conceivably might be involved in this tissue specificity and pathophysiological characteristics. 7607222 7 34 theme [Gal 1097:1100 arg1 alpha- 1119:1124 GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha- 1081:1124 GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha- 1081:1124 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 13 35 theme harmful 1950:1956 arg1 precipitation 1958:1970 the harmful precipitation 1946:1970 the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts 1946:2024 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 5 36 theme human 794:798 arg1 lithostathine 811:823 human pancreatic lithostathine 794:823 human pancreatic lithostathine 794:823 Therefore we isolated the major oligosaccharide chains of human pancreatic lithostathine and determined their sequences by means of NMR analysis. 7607222 9 37 theme common 1214:1219 arg1 core 1238:1241 a common tetrasaccharidic core 1212:1241 a common tetrasaccharidic core 1212:1241 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 12 38 dep electrophoresis 1733:1747 arg1 means 1716:1720 means 1716:1720 means 1716:1720 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 0 39 theme human 21:25 arg1 lithostathine 38:50 human pancreatic lithostathine 21:50 human pancreatic lithostathine 21:50 The glycan moiety of human pancreatic lithostathine. 7607222 12 40 with consistent 1658:1667 arg1 polymorphism 1678:1689 the polymorphism 1674:1689 the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing 1674:1772 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 2 41 theme pancreatic 181:190 arg1 protein 199:205 pancreatic thread protein 181:205 pancreatic thread protein 181:205 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 0 42 theme lithostathine 38:50 arg1 moiety 11:16 The glycan moiety 0:16 The glycan moiety of human pancreatic lithostathine 0:50 The glycan moiety of human pancreatic lithostathine. 7607222 9 43 theme Gal 1244:1246 arg1 GlcNAc 1257:1262 Gal(beta 1-4)GlcNAc 1244:1262 Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)] 1244:1287 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 9 43 theme Gal 1244:1246 arg1 1-6 1269:1271 beta 1-6 1264:1271 beta 1-6 1264:1271 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 11 44 theme sialic 1517:1522 arg1 acid 1524:1527 a sialic acid 1515:1527 a sialic acid in (alpha 2-3) linkage to Gal 1515:1557 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 13 45 theme polypeptide 1990:2000 arg1 precipitation 1958:1970 the harmful precipitation 1946:1970 the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts 1946:2024 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 10 46 theme antigenic 1390:1398 arg1 determinant 1400:1410 the antigenic determinant 1386:1410 the antigenic determinant H 1386:1412 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 2 47 theme regenerating 210:221 arg1 protein 223:229 regenerating protein 210:229 regenerating protein 210:229 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 3 48 theme latter 483:488 arg1 case 490:493 the latter case 479:493 the latter case 479:493 In the latter case, it has been shown recently that it acts as a growth factor which stimulates islet regeneration. 7607222 6 49 theme different 901:909 arg1 glycoforms 911:920 eleven different glycoforms 894:920 eleven different glycoforms 894:920 We obtained eleven different glycoforms and we were able to determine the sequence of seven of them. 7607222 2 50 located found 268:272 arg2 Lithostathine 126:138 Lithostathine 126:138 Lithostathine 126:138 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 2 50 located found 268:272 arg1 pancreas 290:297 the exocrine pancreas 277:297 the exocrine pancreas 277:297 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 2 50 located found 268:272 arg2 glycoprotein 237:248 a glycoprotein 235:248 a glycoprotein which is normally found in the exocrine pancreas 235:297 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 7 51 theme core 1063:1066 arg1 structure 1070:1078 the same core 2 structure 1054:1078 the same core 2 structure 1054:1078 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 13 52 theme Arg11-Ile12 1888:1898 arg1 bond 1900:1903 the Arg11-Ile12 bond 1884:1903 the Arg11-Ile12 bond 1884:1903 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 12 53 theme gel 1729:1731 arg1 electrophoresis 1733:1747 SDS gel electrophoresis 1725:1747 SDS gel electrophoresis 1725:1747 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 5 54 theme major 762:766 arg1 chains 784:789 the major oligosaccharide chains 758:789 the major oligosaccharide chains of human pancreatic lithostathine 758:823 Therefore we isolated the major oligosaccharide chains of human pancreatic lithostathine and determined their sequences by means of NMR analysis. 7607222 10 55 theme Fuc 1441:1443 arg1 presence 1420:1427 presence 1420:1427 presence of external Fuc linked only in (alpha 1-2) to Gal 1420:1477 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 2 56 theme pancreatic 155:164 arg1 protein 172:178 pancreatic stone protein 155:178 pancreatic stone protein 155:178 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 7 57 theme glycosylation 1019:1031 arg1 glycosylation 1019:1031 glycosylation 1019:1031 glycosylation (Thr5) 1019:1038 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 7 57 theme glycosylation 1019:1031 arg1 site 1011:1014 the same site 1002:1014 the same site of glycosylation (Thr5) 1002:1038 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 2 58 theme other 311:315 arg1 tissues 317:323 other tissues 311:323 other tissues 311:323 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 5 59 theme pancreatic 800:809 arg1 lithostathine 811:823 human pancreatic lithostathine 794:823 human pancreatic lithostathine 794:823 Therefore we isolated the major oligosaccharide chains of human pancreatic lithostathine and determined their sequences by means of NMR analysis. 7607222 10 60 attach linked 1445:1450 arg2 alpha 1461:1465 alpha 1-2 1461:1469 alpha 1-2 1461:1469 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 10 60 attach linked 1445:1450 arg2 Fuc 1441:1443 external Fuc 1432:1443 external Fuc linked only in (alpha 1-2) to Gal 1432:1477 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 10 60 attach linked 1445:1450 arg1 Gal 1475:1477 Gal 1475:1477 Gal 1475:1477 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 9 61 theme N-acetyllactosamine 1306:1324 arg1 units 1326:1330 N-acetyllactosamine units 1306:1330 N-acetyllactosamine units 1306:1330 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 8 62 theme sugar 1159:1163 arg1 residues 1165:1172 4 to 9 sugar residues 1152:1172 4 to 9 sugar residues 1152:1172 They ranged in size from 4 to 9 sugar residues. 7607222 8 63 dep 9 1157:1157 arg1 to 1154:1155 to 1154:1155 to 1154:1155 They ranged in size from 4 to 9 sugar residues. 7607222 11 64 theme alpha 1533:1537 arg1 linkage 1544:1550 (alpha 2-3) linkage 1532:1550 (alpha 2-3) linkage to Gal 1532:1557 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 12 65 from form 1796:1799 arg1 consistent 1658:1667 consistent 1658:1667 consistent 1658:1667 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 13 66 theme C-terminal 1979:1988 arg1 polypeptide 1990:2000 the C-terminal polypeptide 1975:2000 the C-terminal polypeptide 1975:2000 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 7 67 gly glycosylation 1019:1031 arg2 glycosylation 1019:1031 glycosylation 1019:1031 glycosylation (Thr5) 1019:1038 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 7 67 gly glycosylation 1019:1031 arg2 site 1011:1014 the same site 1002:1014 the same site of glycosylation (Thr5) 1002:1038 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 12 68 dep focusing 1765:1772 arg1 isoelectric 1753:1763 isoelectric 1753:1763 isoelectric 1753:1763 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 12 69 from consistent 1658:1667 arg1 form 1796:1799 its native form 1785:1799 its native form 1785:1799 These findings are consistent with the polymorphism of the protein, shown by means of SDS gel electrophoresis and isoelectric focusing, either in its native form or after enzymic processing. 7607222 2 70 theme endocrine 455:463 arg1 regeneration 441:452 regeneration 441:452 regeneration (endocrine pancreas) 441:473 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 2 70 theme endocrine 455:463 arg1 pancreas 465:472 endocrine pancreas 455:472 endocrine pancreas 455:472 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 1 71 theme Structure 53:61 arg1 characterization 63:78 Structure characterization 53:78 Structure characterization 53:78 Structure characterization and possible pathophysiological implications. 7607222 9 72 dep Gal 1244:1246 arg1 beta 1248:1251 beta 1-4 1248:1255 beta 1-4 1248:1255 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 7 73 dep [Gal 1097:1100 arg1 1-6 1093:1095 beta 1-6 1088:1095 beta 1-6 1088:1095 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 7 73 dep [Gal 1097:1100 arg1 beta 1102:1105 beta 1-3 1102:1109 beta 1-3 1102:1109 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 4 74 theme pathophysiological 700:717 arg1 characteristics 719:733 pathophysiological characteristics 700:733 pathophysiological characteristics 700:733 Little is known about its glycan moiety, which conceivably might be involved in this tissue specificity and pathophysiological characteristics. 7607222 10 75 dep only 1452:1455 arg1 in 1457:1458 in 1457:1458 in 1457:1458 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 5 76 theme oligosaccharide 768:782 arg1 chains 784:789 the major oligosaccharide chains 758:789 the major oligosaccharide chains of human pancreatic lithostathine 758:823 Therefore we isolated the major oligosaccharide chains of human pancreatic lithostathine and determined their sequences by means of NMR analysis. 7607222 7 77 dep site 1011:1014 arg1 alpha- 1119:1124 GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha- 1081:1124 GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha- 1081:1124 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 7 77 dep site 1011:1014 arg1 all 988:990 all 988:990 all 988:990 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 1 78 theme pathophysiological 93:110 arg1 implications 112:123 possible pathophysiological implications 84:123 possible pathophysiological implications 84:123 Structure characterization and possible pathophysiological implications. 7607222 13 79 from precipitation 1958:1970 arg1 ducts 2020:2024 the pancreatic ducts 2005:2024 the pancreatic ducts 2005:2024 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 7 80 theme beta 1088:1091 arg1 1-6 1093:1095 beta 1-6 1088:1095 beta 1-6 1088:1095 They all were from the same site of glycosylation (Thr5) and displayed the same core 2 structure: GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc alpha-. 7607222 0 81 theme pancreatic 27:36 arg1 lithostathine 38:50 human pancreatic lithostathine 21:50 human pancreatic lithostathine 21:50 The glycan moiety of human pancreatic lithostathine. 7607222 9 82 theme tetrasaccharidic 1221:1236 arg1 core 1238:1241 a common tetrasaccharidic core 1212:1241 a common tetrasaccharidic core 1212:1241 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 2 83 theme thread 192:197 arg1 protein 199:205 pancreatic thread protein 181:205 pancreatic thread protein 181:205 Lithostathine, also known as pancreatic stone protein, pancreatic thread protein or regenerating protein, is a glycoprotein which is normally found in the exocrine pancreas, whereas in other tissues it appears either only under pathological conditions, such as Alzheimer's disease (brain), cancer (colon) or during regeneration (endocrine pancreas). 7607222 11 84 gly disialylated 1569:1580 arg1 form 1582:1585 one disialylated form 1565:1585 one disialylated form which displays a supplementary (alpha 2-6) linkage 1565:1636 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 11 85 theme alpha 1619:1623 arg1 linkage 1630:1636 a supplementary (alpha 2-6) linkage 1602:1636 a supplementary (alpha 2-6) linkage 1602:1636 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 10 86 theme non-reducing 1337:1348 arg1 ends 1350:1353 The non-reducing ends 1333:1353 The non-reducing ends of some oligosaccharides 1333:1378 The non-reducing ends of some oligosaccharides carry the antigenic determinant H, with presence of external Fuc linked only in (alpha 1-2) to Gal. 7607222 11 87 theme supplementary 1604:1616 arg1 linkage 1630:1636 a supplementary (alpha 2-6) linkage 1602:1636 a supplementary (alpha 2-6) linkage 1602:1636 All the glycans, except one, carry a sialic acid in (alpha 2-3) linkage to Gal, with one disialylated form which displays a supplementary (alpha 2-6) linkage. 7607222 13 88 theme in 1910:1911 arg1 hydrolysis 1918:1927 in situ hydrolysis 1910:1927 in situ hydrolysis 1910:1927 Moreover, sialylation seems to protect to some extent the Arg11-Ile12 bond from in situ hydrolysis, thus preventing the harmful precipitation of the C-terminal polypeptide in the pancreatic ducts. 7607222 0 89 theme glycan 4:9 arg1 moiety 11:16 The glycan moiety 0:16 The glycan moiety of human pancreatic lithostathine 0:50 The glycan moiety of human pancreatic lithostathine. 7607222 9 90 theme GlcNAc 1257:1262 arg1 beta 1278:1281 beta 1-3 1278:1285 beta 1-3 1278:1285 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 7607222 9 90 theme GlcNAc 1257:1262 arg1 [Gal 1273:1276 Gal(beta 1-4)GlcNAc(beta 1-6)[Gal 1244:1276 Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)] 1244:1287 Elongation was found to proceed from a common tetrasaccharidic core: Gal(beta 1-4)GlcNAc(beta 1-6)[Gal(beta 1-3)]GalNAc-ol through N-acetyllactosamine units. 1463457 5 0 theme potential 899:907 arg1 sites 925:929 13 potential N-glycosylation sites 896:929 13 potential N-glycosylation sites 896:929 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 6 1 theme arylsulphatase 1170:1183 arg1 family 1185:1190 the arylsulphatase family 1166:1190 the arylsulphatase family 1166:1190 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 2 2 theme recessive 446:454 arg1 disorder 464:471 the autosomal recessive genetic disorder 432:471 the autosomal recessive genetic disorder 432:471 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 4 3 theme derived 716:722 arg1 sequence 735:742 derived amino acid sequence 716:742 derived amino acid sequence 716:742 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 5 4 theme N-glycosylation 909:923 arg1 sites 925:929 13 potential N-glycosylation sites 896:929 13 potential N-glycosylation sites 896:929 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 5 5 theme predicted 804:812 arg1 sequence 814:821 The predicted sequence 800:821 The predicted sequence 800:821 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 1 6 theme heparan 204:210 arg1 sulphate 212:219 heparan sulphate 204:219 heparan sulphate 204:219 Glucosamine-6-sulphatase is an exo-hydrolase required for the lysosomal degradation of heparan sulphate and keratan sulphate. 1463457 7 7 theme substrate 1336:1344 arg1 specificity 1346:1356 substrate specificity 1336:1356 substrate specificity 1336:1356 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 5 8 contain contains 887:894 arg2 sites 925:929 13 potential N-glycosylation sites 896:929 13 potential N-glycosylation sites 896:929 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 5 8 contain contains 887:894 arg1 sequence 814:821 The predicted sequence 800:821 The predicted sequence 800:821 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 2 9 theme sulphate 356:363 arg1 IIID 500:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 4 10 theme acid 730:733 arg1 sequence 735:742 derived amino acid sequence 716:742 derived amino acid sequence 716:742 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 5 11 theme amino 871:875 arg1 acids 877:881 36 amino acids 868:881 36 amino acids 868:881 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 4 12 theme entire 759:764 arg1 protein 791:797 the entire glucosamine-6-sulphatase protein 755:797 the entire glucosamine-6-sulphatase protein 755:797 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 5 13 used used 966:969 arg2 10 959:960 10 959:960 10 959:960 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 2 14 theme heparan 348:354 arg1 sulphate 356:363 heparan sulphate 348:363 heparan sulphate 348:363 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 4 15 theme amino 724:728 arg1 sequence 735:742 derived amino acid sequence 716:742 derived amino acid sequence 716:742 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 5 16 theme acids 877:881 arg1 peptide 857:863 a leader peptide 848:863 a leader peptide of 36 amino acids 848:881 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 6 17 theme other 1033:1037 arg1 sulphatases 1039:1049 other sulphatases 1033:1049 other sulphatases such as the family of arylsulphatases 1033:1087 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 6 17 theme other 1033:1037 arg1 family 1063:1068 the family 1059:1068 the family of arylsulphatases 1059:1087 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 7 18 theme intra-family 1220:1231 arg1 similarity 1233:1242 inter- and intra-family similarity 1209:1242 inter- and intra-family similarity 1209:1242 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 3 19 theme arylsulphatase 592:605 arg1 B 607:607 arylsulphatase B 592:607 arylsulphatase B 592:607 Glucosamine-6-sulphatase can be classified as a non-arylsulphatase since, relative to arylsulphatase B, it shows negligible activity toward 4-methylumbelliferyl sulphate. 1463457 5 20 gly N-glycosylation 909:923 arg2 sites 925:929 13 potential N-glycosylation sites 896:929 13 potential N-glycosylation sites 896:929 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 5 20 gly N-glycosylation 909:923 arg2 13 896:897 13 896:897 13 896:897 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 1 21 theme keratan 225:231 arg1 sulphate 233:240 keratan sulphate 225:240 keratan sulphate 225:240 Glucosamine-6-sulphatase is an exo-hydrolase required for the lysosomal degradation of heparan sulphate and keratan sulphate. 1463457 1 22 theme sulphate 212:219 arg1 degradation 189:199 the lysosomal degradation 175:199 the lysosomal degradation of heparan sulphate and keratan sulphate 175:240 Glucosamine-6-sulphatase is an exo-hydrolase required for the lysosomal degradation of heparan sulphate and keratan sulphate. 1463457 5 23 theme sites 925:929 arg1 likely 947:952 likely 947:952 likely 947:952 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 2 24 theme glycosaminoglycan 329:345 arg1 IIID 500:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 2 25 theme IIID 500:503 arg1 storage 314:320 the lysosomal storage 300:320 the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 300:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 0 26 theme cDNA 2:5 arg1 clone 7:11 A cDNA clone 0:11 A cDNA clone for human glucosamine-6-sulphatase 0:46 A cDNA clone for human glucosamine-6-sulphatase reveals differences between arylsulphatases and non-arylsulphatases. 1463457 3 27 theme 4-methylumbelliferyl 646:665 arg1 sulphate 667:674 4-methylumbelliferyl sulphate 646:674 4-methylumbelliferyl sulphate 646:674 Glucosamine-6-sulphatase can be classified as a non-arylsulphatase since, relative to arylsulphatase B, it shows negligible activity toward 4-methylumbelliferyl sulphate. 1463457 7 28 theme inter- 1209:1214 arg1 similarity 1233:1242 inter- and intra-family similarity 1209:1242 inter- and intra-family similarity 1209:1242 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 6 29 theme similarity 1113:1122 arg1 degree 1103:1108 the degree 1099:1108 the degree of similarity 1099:1122 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 6 29 theme similarity 1113:1122 arg1 high 1134:1137 high 1134:1137 high 1134:1137 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 2 30 theme sulphate 388:395 arg1 6-sulphate 417:426 the monosaccharide sulphate N-acetylglucosamine 6-sulphate 369:426 the monosaccharide sulphate N-acetylglucosamine 6-sulphate 369:426 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 2 31 theme type 495:498 arg1 IIID 500:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 7 32 theme sulphatase 1312:1321 arg1 function 1323:1330 sulphatase function 1312:1330 sulphatase function 1312:1330 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 4 33 theme human 694:698 arg1 clones 705:710 human cDNA clones 694:710 human cDNA clones 694:710 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 2 34 theme monosaccharide 373:386 arg1 6-sulphate 417:426 the monosaccharide sulphate N-acetylglucosamine 6-sulphate 369:426 the monosaccharide sulphate N-acetylglucosamine 6-sulphate 369:426 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 2 35 theme mucopolysaccharidosis 473:493 arg1 IIID 500:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 7 36 theme similarity 1233:1242 arg1 pattern 1198:1204 This pattern 1193:1204 This pattern of inter- and intra-family similarity 1193:1242 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 4 37 theme glucosamine-6-sulphatase 766:789 arg1 protein 791:797 the entire glucosamine-6-sulphatase protein 755:797 the entire glucosamine-6-sulphatase protein 755:797 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 5 38 theme leader 850:855 arg1 peptide 857:863 a leader peptide 848:863 a leader peptide of 36 amino acids 848:881 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 6 39 theme sequence 1010:1017 arg1 similarity 1019:1028 strong sequence similarity 1003:1028 strong sequence similarity to other sulphatases such as the family of arylsulphatases 1003:1087 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 2 40 theme autosomal 436:444 arg1 disorder 464:471 the autosomal recessive genetic disorder 432:471 the autosomal recessive genetic disorder 432:471 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 1 41 theme lysosomal 179:187 arg1 degradation 189:199 the lysosomal degradation 175:199 the lysosomal degradation of heparan sulphate and keratan sulphate 175:240 Glucosamine-6-sulphatase is an exo-hydrolase required for the lysosomal degradation of heparan sulphate and keratan sulphate. 1463457 3 42 theme negligible 619:628 arg1 activity 630:637 negligible activity 619:637 negligible activity toward 4-methylumbelliferyl sulphate 619:674 Glucosamine-6-sulphatase can be classified as a non-arylsulphatase since, relative to arylsulphatase B, it shows negligible activity toward 4-methylumbelliferyl sulphate. 1463457 4 43 theme cDNA 700:703 arg1 clones 705:710 human cDNA clones 694:710 human cDNA clones 694:710 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 6 44 theme strong 1003:1008 arg1 similarity 1019:1028 strong sequence similarity 1003:1028 strong sequence similarity to other sulphatases such as the family of arylsulphatases 1003:1087 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 1 45 theme sulphate 233:240 arg1 degradation 189:199 the lysosomal degradation 175:199 the lysosomal degradation of heparan sulphate and keratan sulphate 175:240 Glucosamine-6-sulphatase is an exo-hydrolase required for the lysosomal degradation of heparan sulphate and keratan sulphate. 1463457 0 46 theme human 17:21 arg1 glucosamine-6-sulphatase 23:46 human glucosamine-6-sulphatase 17:46 human glucosamine-6-sulphatase 17:46 A cDNA clone for human glucosamine-6-sulphatase reveals differences between arylsulphatases and non-arylsulphatases. 1463457 7 47 theme amino 1267:1271 arg1 residues 1278:1285 amino acid residues 1267:1285 amino acid residues 1267:1285 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 6 48 theme arylsulphatases 1073:1087 arg1 family 1063:1068 the family 1059:1068 the family of arylsulphatases 1059:1087 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 5 49 theme amino 831:835 arg1 acids 837:841 552 amino acids 827:841 552 amino acids with a leader peptide of 36 amino acids 827:881 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 2 50 theme glucosamine-6-sulphatase 257:280 arg1 activity 282:289 glucosamine-6-sulphatase activity 257:289 glucosamine-6-sulphatase activity 257:289 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 2 51 theme activity 282:289 arg1 Deficiency 243:252 Deficiency 243:252 Deficiency of glucosamine-6-sulphatase activity 243:289 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 2 52 theme 6-sulphate 417:426 arg1 IIID 500:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 5 53 contain has 823:825 arg1 sequence 814:821 The predicted sequence 800:821 The predicted sequence 800:821 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 5 53 contain has 823:825 arg2 acids 837:841 552 amino acids 827:841 552 amino acids with a leader peptide of 36 amino acids 827:881 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 4 54 link derived 716:722 arg1 sequence 735:742 derived amino acid sequence 716:742 derived amino acid sequence 716:742 We have isolated human cDNA clones and derived amino acid sequence coding for the entire glucosamine-6-sulphatase protein. 1463457 5 55 with acids 837:841 arg1 peptide 857:863 a leader peptide 848:863 a leader peptide of 36 amino acids 848:881 The predicted sequence has 552 amino acids with a leader peptide of 36 amino acids and contains 13 potential N-glycosylation sites, of which it is likely that 10 are used. 1463457 2 56 theme N-acetylglucosamine 397:415 arg1 6-sulphate 417:426 the monosaccharide sulphate N-acetylglucosamine 6-sulphate 369:426 the monosaccharide sulphate N-acetylglucosamine 6-sulphate 369:426 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 6 57 theme family 1185:1190 arg1 members 1155:1161 members 1155:1161 members of the arylsulphatase family 1155:1190 Glucosamine-6-sulphatase shows strong sequence similarity to other sulphatases such as the family of arylsulphatases, although the degree of similarity is not as high as that between members of the arylsulphatase family. 1463457 2 58 theme disorder 464:471 arg1 IIID 500:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 325:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 7 59 theme acid 1273:1276 arg1 residues 1278:1285 amino acid residues 1267:1285 amino acid residues 1267:1285 This pattern of inter- and intra-family similarity delineates regions and amino acid residues that may be critical for sulphatase function and substrate specificity. 1463457 2 60 theme lysosomal 304:312 arg1 storage 314:320 the lysosomal storage 300:320 the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID 300:503 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 1463457 2 61 theme genetic 456:462 arg1 disorder 464:471 the autosomal recessive genetic disorder 432:471 the autosomal recessive genetic disorder 432:471 Deficiency of glucosamine-6-sulphatase activity leads to the lysosomal storage of the glycosaminoglycan, heparan sulphate and the monosaccharide sulphate N-acetylglucosamine 6-sulphate and the autosomal recessive genetic disorder mucopolysaccharidosis type IIID. 10201933 7 0 theme transfectants 1066:1078 arg1 analyses 1038:1045 Flow cytometric and immunoprecipitation analyses 998:1045 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants 998:1078 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 6 1 theme membrane-anchoring 973:990 arg1 form 992:995 the membrane-anchoring form 969:995 the membrane-anchoring form 969:995 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 1 2 theme 80-kDa 150:155 arg1 glycoprotein 210:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein 147:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E 147:285 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 1 2 theme 80-kDa 150:155 arg1 CDw108 78:83 CDw108 78:83 CDw108 78:83 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 4 3 theme cDNA 593:596 arg1 frame 573:577 the open reading frame 556:577 the open reading frame of the cloned cDNA 556:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 10 4 theme Ag 1536:1537 arg1 function 1513:1520 the biological function 1498:1520 the biological function of the CDw108 Ag 1498:1537 This molecular information will be critical for understanding the biological function of the CDw108 Ag. 10201933 4 5 theme 1998-base 537:545 arg1 pairs 547:551 The 1998-base pairs 533:551 The 1998-base pairs of the open reading frame of the cloned cDNA 533:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 5 theme 1998-base 537:545 arg1 protein 608:614 a protein 606:614 a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif 606:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 9 6 theme CDw108 1353:1358 arg1 15q23-24 1425:1432 15q23-24 1425:1432 15q23-24 1425:1432 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 6 theme CDw108 1353:1358 arg1 located 1369:1375 located 1369:1375 located 1369:1375 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 6 theme CDw108 1353:1358 arg1 gene 1360:1363 the CDw108 gene 1349:1363 the CDw108 gene 1349:1363 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 5 7 theme form 837:840 arg1 kDa 849:851 68 kDa 846:851 68 kDa 846:851 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 5 7 theme form 837:840 arg1 mass 810:813 the estimated molecular mass 786:813 the estimated molecular mass of the unglycosylated form 786:840 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 7 8 theme cDNA 1061:1064 arg1 transfectants 1066:1078 the CDw108 cDNA transfectants 1050:1078 the CDw108 cDNA transfectants 1050:1078 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 8 9 theme activated 1179:1187 arg1 PBMCs 1189:1193 activated PBMCs 1179:1193 activated PBMCs 1179:1193 The CDw108 mRNA was expressed in activated PBMCs as well as in the spleen, thymus, testis, placenta, and brain, but was not expressed in any other tissues tested. 10201933 4 10 theme cloned 586:591 arg1 cDNA 593:596 the cloned cDNA 582:596 the cloned cDNA 582:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 9 11 theme arm 1403:1405 arg1 middle 1384:1389 the middle 1380:1389 the middle of the long arm of chromosome 15 1380:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 2 12 theme biological 322:331 arg1 function 333:340 biological function 322:340 biological function 322:340 The molecular characteristics and biological function of the CDw108 were not clarified previously. 10201933 9 13 theme hybrid 1319:1324 arg1 mapping 1326:1332 Radiation hybrid mapping 1309:1332 Radiation hybrid mapping 1309:1332 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 10 14 theme molecular 1441:1449 arg1 information 1451:1461 This molecular information 1436:1461 This molecular information 1436:1461 This molecular information will be critical for understanding the biological function of the CDw108 Ag. 10201933 4 15 theme motif 715:719 arg1 aa 658:659 the 46 aa 651:659 the 46 aa of the signal peptide 651:681 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 15 theme motif 715:719 arg1 aa 694:695 the 19 aa 687:695 the 19 aa of the GPI-anchor motif 687:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 3 16 contain containing 434:443 arg2 sequence 463:470 the entire coding sequence 445:470 the entire coding sequence of the CDw108 gene 445:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 3 16 contain containing 434:443 arg1 clone 428:432 the cDNA clone 419:432 the cDNA clone containing the entire coding sequence of the CDw108 gene 419:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 4 17 theme signal 668:673 arg1 peptide 675:681 the signal peptide 664:681 the signal peptide 664:681 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 5 18 theme estimated 790:798 arg1 kDa 849:851 68 kDa 846:851 68 kDa 846:851 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 5 18 theme estimated 790:798 arg1 mass 810:813 the estimated molecular mass 786:813 the estimated molecular mass of the unglycosylated form 786:840 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 1 19 gly glycoprotein 210:221 arg1 glycoprotein 210:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein 147:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E 147:285 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 1 19 gly glycoprotein 210:221 arg1 CDw108 78:83 CDw108 78:83 CDw108 78:83 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 4 20 theme acids 629:633 arg1 pairs 547:551 The 1998-base pairs 533:551 The 1998-base pairs of the open reading frame of the cloned cDNA 533:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 20 theme acids 629:633 arg1 protein 608:614 a protein 606:614 a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif 606:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 2 21 theme molecular 292:300 arg1 characteristics 302:316 The molecular characteristics 288:316 The molecular characteristics 288:316 The molecular characteristics and biological function of the CDw108 were not clarified previously. 10201933 3 22 theme coding 456:461 arg1 sequence 463:470 the entire coding sequence 445:470 the entire coding sequence of the CDw108 gene 445:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 0 23 theme CDw108 70:75 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108. 0:76 Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108. 10201933 6 24 theme N-linked 924:931 arg1 sites 947:951 the five potential N-linked glycosylation sites 905:951 the five potential N-linked glycosylation sites 905:951 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 24 theme N-linked 924:931 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 5 25 theme molecular 800:808 arg1 kDa 849:851 68 kDa 846:851 68 kDa 846:851 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 5 25 theme molecular 800:808 arg1 mass 810:813 the estimated molecular mass 786:813 the estimated molecular mass of the unglycosylated form 786:840 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 5 26 theme CDw108 759:764 arg1 form 751:754 the membrane-anchoring form 728:754 the membrane-anchoring form of CDw108 728:764 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 5 26 theme CDw108 759:764 arg1 aa 778:779 the 602 aa 770:779 the 602 aa 770:779 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 6 27 theme cell 876:879 arg1 sequence 892:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 27 theme cell 876:879 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 7 28 theme immunoprecipitation 1018:1036 arg1 analyses 1038:1045 Flow cytometric and immunoprecipitation analyses 998:1045 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants 998:1078 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 7 29 theme native 1123:1128 arg1 cDNA 1106:1109 the cloned cDNA 1095:1109 the cloned cDNA 1095:1109 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 7 29 theme native 1123:1128 arg1 form 1130:1133 the native form 1119:1133 the native form of CDw108 1119:1143 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 0 30 theme Molecular 0:8 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108. 0:76 Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108. 10201933 4 31 theme frame 573:577 arg1 pairs 547:551 The 1998-base pairs 533:551 The 1998-base pairs of the open reading frame of the cloned cDNA 533:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 31 theme frame 573:577 arg1 protein 608:614 a protein 606:614 a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif 606:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 7 32 theme cloned 1099:1104 arg1 cDNA 1106:1109 the cloned cDNA 1095:1109 the cloned cDNA 1095:1109 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 7 32 theme cloned 1099:1104 arg1 form 1130:1133 the native form 1119:1133 the native form of CDw108 1119:1143 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 6 33 located located 958:964 arg2 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 33 located located 958:964 arg2 sites 947:951 the five potential N-linked glycosylation sites 905:951 the five potential N-linked glycosylation sites 905:951 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 33 located located 958:964 arg1 form 992:995 the membrane-anchoring form 969:995 the membrane-anchoring form 969:995 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 33 located located 958:964 arg2 sequence 892:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 3 34 theme entire 449:454 arg1 sequence 463:470 the entire coding sequence 445:470 the entire coding sequence of the CDw108 gene 445:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 4 35 theme reading 565:571 arg1 frame 573:577 the open reading frame 556:577 the open reading frame of the cloned cDNA 556:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 7 36 theme cytometric 1003:1012 arg1 analyses 1038:1045 Flow cytometric and immunoprecipitation analyses 998:1045 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants 998:1078 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 9 37 from middle 1384:1389 arg1 15q23-24 1425:1432 15q23-24 1425:1432 15q23-24 1425:1432 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 37 from middle 1384:1389 arg1 located 1369:1375 located 1369:1375 located 1369:1375 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 37 from middle 1384:1389 arg1 gene 1360:1363 the CDw108 gene 1349:1363 the CDw108 gene 1349:1363 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 1 38 theme John-Milton-Hagen 104:120 arg1 Ag 140:141 the John-Milton-Hagen human blood group Ag 100:141 the John-Milton-Hagen human blood group Ag 100:141 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 3 39 theme CDw108 479:484 arg1 gene 486:489 the CDw108 gene 475:489 the CDw108 gene 475:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 6 40 theme attachment 881:890 arg1 sequence 892:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 40 theme attachment 881:890 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 1 41 theme human 122:126 arg1 Ag 140:141 the John-Milton-Hagen human blood group Ag 100:141 the John-Milton-Hagen human blood group Ag 100:141 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 9 42 theme chromosome 1410:1419 arg1 arm 1403:1405 the long arm 1394:1405 the long arm of chromosome 15 1394:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 42 theme chromosome 1410:1419 arg1 chromosome 1410:1419 chromosome 15 1410:1422 chromosome 15 1410:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 3 43 theme gene 486:489 arg1 sequence 463:470 the entire coding sequence 445:470 the entire coding sequence of the CDw108 gene 445:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 0 44 theme molecule 61:68 arg1 CDw108 70:75 a glycosylphosphatidylinositol-anchored molecule CDw108 21:75 a glycosylphosphatidylinositol-anchored molecule CDw108 21:75 Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108. 10201933 3 45 theme molecular 506:514 arg1 characteristics 516:530 its molecular characteristics 502:530 its molecular characteristics 502:530 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 8 46 theme other 1287:1291 arg1 tissues 1293:1299 any other tissues 1283:1299 any other tissues tested 1283:1306 The CDw108 mRNA was expressed in activated PBMCs as well as in the spleen, thymus, testis, placenta, and brain, but was not expressed in any other tissues tested. 10201933 1 47 theme -anchored 191:199 arg1 glycoprotein 210:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein 147:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E 147:285 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 1 47 theme -anchored 191:199 arg1 CDw108 78:83 CDw108 78:83 CDw108 78:83 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 0 48 theme glycosylphosphatidylinositol-anchored 23:59 arg1 CDw108 70:75 a glycosylphosphatidylinositol-anchored molecule CDw108 21:75 a glycosylphosphatidylinositol-anchored molecule CDw108 21:75 Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108. 10201933 5 49 theme membrane-anchoring 732:749 arg1 form 751:754 the membrane-anchoring form 728:754 the membrane-anchoring form of CDw108 728:764 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 5 49 theme membrane-anchoring 732:749 arg1 aa 778:779 the 602 aa 770:779 the 602 aa 770:779 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 4 50 theme peptide 675:681 arg1 aa 658:659 the 46 aa 651:659 the 46 aa of the signal peptide 651:681 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 50 theme peptide 675:681 arg1 aa 694:695 the 19 aa 687:695 the 19 aa of the GPI-anchor motif 687:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 9 51 from located 1369:1375 arg1 middle 1384:1389 the middle 1380:1389 the middle of the long arm of chromosome 15 1380:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 7 52 theme CDw108 1138:1143 arg1 cDNA 1106:1109 the cloned cDNA 1095:1109 the cloned cDNA 1095:1109 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 7 52 theme CDw108 1138:1143 arg1 form 1130:1133 the native form 1119:1133 the native form of CDw108 1119:1143 Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. 10201933 6 53 theme Arg-Gly-Asp 863:873 arg1 sequence 892:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 53 theme Arg-Gly-Asp 863:873 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 5 54 gly unglycosylated 822:835 arg1 form 837:840 the unglycosylated form 818:840 the unglycosylated form 818:840 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 9 55 located located 1369:1375 arg2 15q23-24 1425:1432 15q23-24 1425:1432 15q23-24 1425:1432 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 55 located located 1369:1375 arg2 located 1369:1375 located 1369:1375 located 1369:1375 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 55 located located 1369:1375 arg1 middle 1384:1389 the middle 1380:1389 the middle of the long arm of chromosome 15 1380:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 55 located located 1369:1375 arg2 gene 1360:1363 the CDw108 gene 1349:1363 the CDw108 gene 1349:1363 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 1 56 theme blood 128:132 arg1 Ag 140:141 the John-Milton-Hagen human blood group Ag 100:141 the John-Milton-Hagen human blood group Ag 100:141 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 3 57 theme cDNA 423:426 arg1 clone 428:432 the cDNA clone 419:432 the cDNA clone containing the entire coding sequence of the CDw108 gene 419:489 In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. 10201933 4 58 theme open 560:563 arg1 frame 573:577 the open reading frame 556:577 the open reading frame of the cloned cDNA 556:596 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 1 59 theme group 134:138 arg1 Ag 140:141 the John-Milton-Hagen human blood group Ag 100:141 the John-Milton-Hagen human blood group Ag 100:141 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 6 60 theme RGD 858:860 arg1 sequence 892:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence 854:899 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 60 theme RGD 858:860 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 9 61 dep located 1369:1375 arg1 15q23-24 1425:1432 15q23-24 1425:1432 15q23-24 1425:1432 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 61 dep located 1369:1375 arg1 located 1369:1375 located 1369:1375 located 1369:1375 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 61 dep located 1369:1375 arg1 gene 1360:1363 the CDw108 gene 1349:1363 the CDw108 gene 1349:1363 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 6 62 link N-linked 924:931 arg1 sites 947:951 the five potential N-linked glycosylation sites 905:951 the five potential N-linked glycosylation sites 905:951 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 62 link N-linked 924:931 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 4 63 theme GPI-anchor 704:713 arg1 motif 715:719 the GPI-anchor motif 700:719 the GPI-anchor motif 700:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 9 64 theme Radiation 1309:1317 arg1 mapping 1326:1332 Radiation hybrid mapping 1309:1332 Radiation hybrid mapping 1309:1332 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 2 65 theme CDw108 349:354 arg1 characteristics 302:316 The molecular characteristics 288:316 The molecular characteristics 288:316 The molecular characteristics and biological function of the CDw108 were not clarified previously. 10201933 2 65 theme CDw108 349:354 arg1 function 333:340 biological function 322:340 biological function 322:340 The molecular characteristics and biological function of the CDw108 were not clarified previously. 10201933 1 66 theme membrane 201:208 arg1 glycoprotein 210:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein 147:221 an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E 147:285 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 1 66 theme membrane 201:208 arg1 CDw108 78:83 CDw108 78:83 CDw108 78:83 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10201933 9 67 theme long 1398:1401 arg1 arm 1403:1405 the long arm 1394:1405 the long arm of chromosome 15 1394:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 9 67 theme long 1398:1401 arg1 chromosome 1410:1419 chromosome 15 1410:1422 chromosome 15 1410:1422 Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). 10201933 6 68 theme potential 914:922 arg1 sites 947:951 the five potential N-linked glycosylation sites 905:951 the five potential N-linked glycosylation sites 905:951 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 68 theme potential 914:922 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 4 69 theme amino 623:627 arg1 aa 636:637 aa 636:637 aa 636:637 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 69 theme amino 623:627 arg1 acids 629:633 666 amino acids 619:633 666 amino acids (aa) 619:638 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 69 theme amino 623:627 arg1 aa 694:695 the 19 aa 687:695 the 19 aa of the GPI-anchor motif 687:719 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 4 69 theme amino 623:627 arg1 aa 658:659 the 46 aa 651:659 the 46 aa of the signal peptide 651:681 The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. 10201933 6 70 theme glycosylation 933:945 arg1 sites 947:951 the five potential N-linked glycosylation sites 905:951 the five potential N-linked glycosylation sites 905:951 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 70 theme glycosylation 933:945 arg1 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 71 gly glycosylation 933:945 arg2 located 958:964 located 958:964 located 958:964 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 71 gly glycosylation 933:945 arg2 sites 947:951 the five potential N-linked glycosylation sites 905:951 the five potential N-linked glycosylation sites 905:951 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 71 gly glycosylation 933:945 arg2 form 992:995 the membrane-anchoring form 969:995 the membrane-anchoring form 969:995 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 6 71 gly glycosylation 933:945 arg2 five 909:912 five 909:912 five 909:912 The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. 10201933 5 72 theme unglycosylated 822:835 arg1 form 837:840 the unglycosylated form 818:840 the unglycosylated form 818:840 Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. 10201933 10 73 theme biological 1502:1511 arg1 function 1513:1520 the biological function 1498:1520 the biological function of the CDw108 Ag 1498:1537 This molecular information will be critical for understanding the biological function of the CDw108 Ag. 10201933 1 74 theme activated 259:267 arg1 lymphocytes 269:279 activated lymphocytes 259:279 activated lymphocytes 259:279 CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. 10397151 12 0 dep A 1572:1572 arg1 = 1577:1577 = 1577:1577 = 1577:1577 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 10 1 theme mannose 1308:1314 arg1 chains 1337:1342 core mannose oligosaccharide side chains 1303:1342 core mannose oligosaccharide side chains 1303:1342 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 1 theme mannose 1308:1314 arg1 N-glycans 1252:1260 the N-glycans 1248:1260 the N-glycans of baculovirus derived Fc gamma RIIa 1248:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 7 2 theme Fc 940:941 arg1 RIIa 949:952 derived Fc gamma RIIa 932:952 derived Fc gamma RIIa 932:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 9 3 used occupied 1175:1182 arg2 sites 1165:1169 both potential N-linked glycosylation sites 1127:1169 both potential N-linked glycosylation sites 1127:1169 Furthermore, both potential N-linked glycosylation sites are occupied. 10397151 4 4 theme recombinant 535:545 arg1 RIIa 556:559 recombinant Fc gamma RIIa 535:559 recombinant Fc gamma RIIa that has been used to generate high quality crystals 535:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 7 5 theme cell 927:930 arg1 analysis 894:901 Biochemical analysis 882:901 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa 882:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 2 6 theme IgG 237:239 arg1 isotypes 241:248 certain IgG isotypes 229:248 certain IgG isotypes 229:248 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 6 7 theme gamma 766:770 arg1 R 772:772 other Fc gamma R 757:772 other Fc gamma R 757:772 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 4 8 theme high 592:595 arg1 crystals 605:612 high quality crystals 592:612 high quality crystals 592:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 11 9 theme RIIa 1444:1447 arg1 crystals 1415:1422 diffraction quality crystals 1395:1422 diffraction quality crystals of soluble Fc gamma RIIa 1395:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 10 10 theme mass 1209:1212 arg1 ESMS 1228:1231 ESMS 1228:1231 ESMS 1228:1231 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 10 theme mass 1209:1212 arg1 spectrometry 1214:1225 Electrospray ionisation mass spectrometry 1185:1225 Electrospray ionisation mass spectrometry (ESMS) 1185:1232 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 1 11 theme specific 130:137 arg1 receptors 152:160 specific cell surface receptors 130:160 specific cell surface receptors for immunoglobulin 130:179 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 6 12 theme other 757:761 arg1 R 772:772 other Fc gamma R 757:772 other Fc gamma R 757:772 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 12 13 theme space 1519:1523 arg1 P2 1531:1532 the space group P2(1)2(1)2 1515:1540 the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A 1515:1597 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 7 14 dep cell 927:930 arg1 RIIa 949:952 derived Fc gamma RIIa 932:952 derived Fc gamma RIIa 932:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 10 15 theme Electrospray 1185:1196 arg1 ESMS 1228:1231 ESMS 1228:1231 ESMS 1228:1231 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 15 theme Electrospray 1185:1196 arg1 spectrometry 1214:1225 Electrospray ionisation mass spectrometry 1185:1225 Electrospray ionisation mass spectrometry (ESMS) 1185:1232 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 2 16 theme important 257:265 arg1 roles 267:271 important roles 257:271 important roles 257:271 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 1 17 theme surface 144:150 arg1 receptors 152:160 specific cell surface receptors 130:160 specific cell surface receptors for immunoglobulin 130:179 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 7 18 theme Biochemical 882:892 arg1 analysis 894:901 Biochemical analysis 882:901 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa 882:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 5 19 theme gamma 654:658 arg1 RII 660:662 the Fc gamma RII 647:662 the Fc gamma RII 647:662 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 7 20 theme genuine 970:976 arg1 N-terminus 978:987 the genuine N-terminus 966:987 the genuine N-terminus 966:987 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 6 21 theme gamma 797:801 arg1 K 809:809 K 809:809 K(D) = 3 microM 809:823 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 21 theme gamma 797:801 arg1 RIIa 803:806 Fc gamma RIIa 794:806 Fc gamma RIIa 794:806 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 12 22 from resolution 1501:1510 arg1 P2 1531:1532 the space group P2(1)2(1)2 1515:1540 the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A 1515:1597 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 5 23 theme Equilibrium 615:625 arg1 analysis 635:642 Equilibrium binding analysis 615:642 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 615:690 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 2 24 theme Fc 182:183 arg1 RIIa 191:194 Fc gamma RIIa 182:194 Fc gamma RIIa 182:194 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 1 25 theme gamma 97:101 arg1 RIIa 103:106 Fc gamma RIIa 94:106 Fc gamma RIIa 94:106 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 2 26 theme immune 209:214 arg1 complexes 216:224 immune complexes 209:224 immune complexes of certain IgG isotypes 209:248 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 4 27 used used 575:578 arg2 RIIa 556:559 recombinant Fc gamma RIIa 535:559 recombinant Fc gamma RIIa that has been used to generate high quality crystals 535:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 7 28 theme sequence 1030:1037 arg1 Q 1039:1039 the sequence Q 1026:1039 the sequence Q 1026:1039 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 12 29 with P2 1531:1532 arg1 dimensions 1552:1561 cell dimensions 1547:1561 cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A 1547:1597 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 0 30 theme low 63:65 arg1 receptor 76:83 the low affinity receptor 59:83 the low affinity receptor for IgG 59:91 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 0 30 theme low 63:65 arg1 RIIa 53:56 Fc gamma RIIa 44:56 Fc gamma RIIa 44:56 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 13 31 theme Fc 1770:1771 arg1 RIIa 1779:1782 Fc gamma RIIa 1770:1782 Fc gamma RIIa 1770:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 1 32 theme family 120:125 arg1 one 111:113 one 111:113 one 111:113 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 1 32 theme family 120:125 arg1 family 120:125 a family 118:125 a family of specific cell surface receptors for immunoglobulin 118:179 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 13 33 with identity 1756:1763 arg1 RIIa 1779:1782 Fc gamma RIIa 1770:1782 Fc gamma RIIa 1770:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 7 34 theme amino 1012:1016 arg1 A 1042:1042 A 1042:1042 A 1042:1042 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 34 theme amino 1012:1016 arg1 P 1051:1051 P 1051:1051 P 1051:1051 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 34 theme amino 1012:1016 arg1 acid 1018:1021 the first amino acid 1002:1021 the first amino acid in the sequence Q 1002:1039 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 34 theme amino 1012:1016 arg1 Q 994:994 Q 994:994 Q 994:994 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 34 theme amino 1012:1016 arg1 A 1045:1045 A 1045:1045 A 1045:1045 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 34 theme amino 1012:1016 arg1 A 1048:1048 A 1048:1048 A 1048:1048 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 3 35 theme RIIa 392:395 arg1 characteristics 317:331 the precise characteristics 305:331 the precise characteristics of IgG binding 305:346 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 3 35 theme RIIa 392:395 arg1 structure 370:378 three-dimensional structure 352:378 three-dimensional structure of Fc gamma RIIa 352:395 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 13 36 theme amino 1745:1749 arg1 identity 1756:1763 high amino acid identity 1740:1763 high amino acid identity with Fc gamma RIIa 1740:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 13 37 theme Fc 1690:1691 arg1 RIIa 1699:1702 Fc gamma RIIa 1690:1702 Fc gamma RIIa 1690:1702 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 3 38 theme IgG 336:338 arg1 binding 340:346 IgG binding 336:346 IgG binding 336:346 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 10 39 link derived 1277:1283 arg1 RIIa 1294:1297 baculovirus derived Fc gamma RIIa 1265:1297 baculovirus derived Fc gamma RIIa 1265:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 3 40 theme Fc 383:384 arg1 RIIa 392:395 Fc gamma RIIa 383:395 Fc gamma RIIa 383:395 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 0 41 theme Biochemical 0:10 arg1 analysis 12:19 Biochemical analysis 0:19 Biochemical analysis 0:19 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 6 42 theme IgG4 867:870 arg1 receptor 872:879 an IgG4 receptor 864:879 an IgG4 receptor 864:879 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 42 theme IgG4 867:870 arg1 RIIa 856:859 Fc gamma RIIa 847:859 Fc gamma RIIa 847:859 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 4 43 theme interaction 480:490 arg1 characterisation 515:530 biochemical characterisation 503:530 the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals 442:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 4 43 theme interaction 480:490 arg1 affinity 446:453 the affinity 442:453 the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals 442:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 10 44 theme Fc 1285:1286 arg1 RIIa 1294:1297 baculovirus derived Fc gamma RIIa 1265:1297 baculovirus derived Fc gamma RIIa 1265:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 45 theme baculovirus 1265:1275 arg1 RIIa 1294:1297 baculovirus derived Fc gamma RIIa 1265:1297 baculovirus derived Fc gamma RIIa 1265:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 7 46 link derived 932:938 arg1 RIIa 949:952 derived Fc gamma RIIa 932:952 derived Fc gamma RIIa 932:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 6 47 theme gamma 850:854 arg1 receptor 872:879 an IgG4 receptor 864:879 an IgG4 receptor 864:879 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 47 theme gamma 850:854 arg1 RIIa 856:859 Fc gamma RIIa 847:859 Fc gamma RIIa 847:859 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 4 48 theme gamma 465:469 arg1 RIIa 471:474 Fc gamma RIIa 462:474 the Fc gamma RIIa:IgG interaction 458:490 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 10 49 theme core 1303:1306 arg1 chains 1337:1342 core mannose oligosaccharide side chains 1303:1342 core mannose oligosaccharide side chains 1303:1342 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 49 theme core 1303:1306 arg1 N-glycans 1252:1260 the N-glycans 1248:1260 the N-glycans of baculovirus derived Fc gamma RIIa 1248:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 13 50 theme FcR 1716:1718 arg1 proteins 1720:1727 related FcR proteins 1708:1727 related FcR proteins that share high amino acid identity with Fc gamma RIIa 1708:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 0 51 theme gamma 47:51 arg1 receptor 76:83 the low affinity receptor 59:83 the low affinity receptor for IgG 59:91 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 0 51 theme gamma 47:51 arg1 RIIa 53:56 Fc gamma RIIa 44:56 Fc gamma RIIa 44:56 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 9 52 theme potential 1132:1140 arg1 sites 1165:1169 both potential N-linked glycosylation sites 1127:1169 both potential N-linked glycosylation sites 1127:1169 Furthermore, both potential N-linked glycosylation sites are occupied. 10397151 12 53 theme 78.8 1567:1570 arg1 A 1572:1572 a = 78.8 A 1563:1572 a = 78.8 A 1563:1572 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 12 54 theme = 1565:1565 arg1 A 1572:1572 a = 78.8 A 1563:1572 a = 78.8 A 1563:1572 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 10 55 theme RIIa 1294:1297 arg1 chains 1337:1342 core mannose oligosaccharide side chains 1303:1342 core mannose oligosaccharide side chains 1303:1342 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 55 theme RIIa 1294:1297 arg1 N-glycans 1252:1260 the N-glycans 1248:1260 the N-glycans of baculovirus derived Fc gamma RIIa 1248:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 2 56 theme immune 276:281 arg1 homeostasis 283:293 immune homeostasis 276:293 immune homeostasis 276:293 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 9 57 theme glycosylation 1151:1163 arg1 sites 1165:1169 both potential N-linked glycosylation sites 1127:1169 both potential N-linked glycosylation sites 1127:1169 Furthermore, both potential N-linked glycosylation sites are occupied. 10397151 13 58 theme related 1708:1714 arg1 proteins 1720:1727 related FcR proteins 1708:1727 related FcR proteins that share high amino acid identity with Fc gamma RIIa 1708:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 4 59 theme Fc 547:548 arg1 RIIa 556:559 recombinant Fc gamma RIIa 535:559 recombinant Fc gamma RIIa that has been used to generate high quality crystals 535:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 11 60 theme crystals 1415:1422 arg1 crystallisation 1376:1390 the first crystallisation 1366:1390 the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa 1366:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 0 61 theme RIIa 53:56 arg1 crystallisation 25:39 crystallisation 25:39 crystallisation 25:39 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 0 61 theme RIIa 53:56 arg1 analysis 12:19 Biochemical analysis 0:19 Biochemical analysis 0:19 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 4 62 theme RIIa 556:559 arg1 characterisation 515:530 biochemical characterisation 503:530 the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals 442:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 4 62 theme RIIa 556:559 arg1 affinity 446:453 the affinity 442:453 the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals 442:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 10 63 theme oligosaccharide 1316:1330 arg1 chains 1337:1342 core mannose oligosaccharide side chains 1303:1342 core mannose oligosaccharide side chains 1303:1342 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 63 theme oligosaccharide 1316:1330 arg1 N-glycans 1252:1260 the N-glycans 1248:1260 the N-glycans of baculovirus derived Fc gamma RIIa 1248:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 11 64 theme gamma 1438:1442 arg1 RIIa 1444:1447 soluble Fc gamma RIIa 1427:1447 soluble Fc gamma RIIa 1427:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 7 65 theme derived 932:938 arg1 RIIa 949:952 derived Fc gamma RIIa 932:952 derived Fc gamma RIIa 932:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 11 66 theme diffraction 1395:1405 arg1 crystals 1415:1422 diffraction quality crystals 1395:1422 diffraction quality crystals of soluble Fc gamma RIIa 1395:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 4 67 theme biochemical 503:513 arg1 characterisation 515:530 biochemical characterisation 503:530 the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals 442:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 11 68 theme soluble 1427:1433 arg1 RIIa 1444:1447 soluble Fc gamma RIIa 1427:1447 soluble Fc gamma RIIa 1427:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 7 69 theme insect 920:925 arg1 analysis 894:901 Biochemical analysis 882:901 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa 882:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 13 70 theme three-dimensional 1659:1675 arg1 structure 1677:1685 the three-dimensional structure 1655:1685 the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa 1655:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 2 71 theme isotypes 241:248 arg1 complexes 216:224 immune complexes 209:224 immune complexes of certain IgG isotypes 209:248 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 12 72 theme cell 1547:1550 arg1 dimensions 1552:1561 cell dimensions 1547:1561 cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A 1547:1597 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 7 73 theme mammalian 906:914 arg1 cell 927:930 mammalian and insect cell derived Fc gamma RIIa 906:952 cell 927:930 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 2 74 theme certain 229:235 arg1 isotypes 241:248 certain IgG isotypes 229:248 certain IgG isotypes 229:248 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 6 75 theme Fc 763:764 arg1 R 772:772 other Fc gamma R 757:772 other Fc gamma R 757:772 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 12 76 theme group 1525:1529 arg1 P2 1531:1532 the space group P2(1)2(1)2 1515:1540 the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A 1515:1597 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 10 77 theme ionisation 1198:1207 arg1 ESMS 1228:1231 ESMS 1228:1231 ESMS 1228:1231 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 77 theme ionisation 1198:1207 arg1 spectrometry 1214:1225 Electrospray ionisation mass spectrometry 1185:1225 Electrospray ionisation mass spectrometry (ESMS) 1185:1232 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 1 78 theme cell 139:142 arg1 receptors 152:160 specific cell surface receptors 130:160 specific cell surface receptors for immunoglobulin 130:179 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 4 79 theme quality 597:603 arg1 crystals 605:612 high quality crystals 592:612 high quality crystals 592:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 1 80 theme receptors 152:160 arg1 family 120:125 a family 118:125 a family of specific cell surface receptors for immunoglobulin 118:179 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 5 81 theme Fc 651:652 arg1 RII 660:662 the Fc gamma RII 647:662 the Fc gamma RII 647:662 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 13 82 dep significant 1613:1623 arg1 advance 1625:1631 advance 1625:1631 advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa 1625:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 5 83 theme microM 729:734 arg1 affinity 706:713 an affinity 703:713 an affinity of K(D) = 0.6 microM 703:734 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 7 84 theme first 1006:1010 arg1 A 1042:1042 A 1042:1042 A 1042:1042 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 84 theme first 1006:1010 arg1 P 1051:1051 P 1051:1051 P 1051:1051 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 84 theme first 1006:1010 arg1 acid 1018:1021 the first amino acid 1002:1021 the first amino acid in the sequence Q 1002:1039 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 84 theme first 1006:1010 arg1 Q 994:994 Q 994:994 Q 994:994 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 84 theme first 1006:1010 arg1 A 1045:1045 A 1045:1045 A 1045:1045 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 7 84 theme first 1006:1010 arg1 A 1048:1048 A 1048:1048 A 1048:1048 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 2 85 theme gamma 185:189 arg1 RIIa 191:194 Fc gamma RIIa 182:194 Fc gamma RIIa 182:194 Fc gamma RIIa, which binds immune complexes of certain IgG isotypes, plays important roles in immune homeostasis. 10397151 1 86 theme Fc 94:95 arg1 RIIa 103:106 Fc gamma RIIa 94:106 Fc gamma RIIa 94:106 Fc gamma RIIa is one of a family of specific cell surface receptors for immunoglobulin. 10397151 6 87 theme Fc 794:795 arg1 K 809:809 K 809:809 K(D) = 3 microM 809:823 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 87 theme Fc 794:795 arg1 RIIa 803:806 Fc gamma RIIa 794:806 Fc gamma RIIa 794:806 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 7 88 from acid 1018:1021 arg1 Q 1039:1039 the sequence Q 1026:1039 the sequence Q 1026:1039 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 5 89 theme binding 627:633 arg1 analysis 635:642 Equilibrium binding analysis 615:642 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 615:690 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 6 90 theme Fc 847:848 arg1 receptor 872:879 an IgG4 receptor 864:879 an IgG4 receptor 864:879 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 90 theme Fc 847:848 arg1 RIIa 856:859 Fc gamma RIIa 847:859 Fc gamma RIIa 847:859 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 5 91 theme = 723:723 arg1 microM 729:734 K(D) = 0.6 microM 718:734 K(D) = 0.6 microM 718:734 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 6 92 dep = 814:814 arg1 microM 818:823 3 microM 816:823 3 microM 816:823 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 5 93 dep analysis 635:642 arg1 interaction 668:678 IgG interaction 664:678 IgG interaction found 664:684 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 5 93 dep analysis 635:642 arg1 IgG3 687:690 IgG3 687:690 IgG3 687:690 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 0 94 theme affinity 67:74 arg1 receptor 76:83 the low affinity receptor 59:83 the low affinity receptor for IgG 59:91 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 0 94 theme affinity 67:74 arg1 RIIa 53:56 Fc gamma RIIa 44:56 Fc gamma RIIa 44:56 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 4 95 theme Fc 462:463 arg1 RIIa 471:474 Fc gamma RIIa 462:474 the Fc gamma RIIa:IgG interaction 458:490 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 5 96 theme RII 660:662 arg1 analysis 635:642 Equilibrium binding analysis 615:642 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 615:690 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 13 97 theme gamma 1773:1777 arg1 RIIa 1779:1782 Fc gamma RIIa 1770:1782 Fc gamma RIIa 1770:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 6 98 theme = 814:814 arg1 K 809:809 K 809:809 K(D) = 3 microM 809:823 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 98 theme = 814:814 arg1 RIIa 803:806 Fc gamma RIIa 794:806 Fc gamma RIIa 794:806 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 6 98 theme = 814:814 arg1 D 811:811 D 811:811 D 811:811 Unlike other Fc gamma R, IgG4 also bound to Fc gamma RIIa, K(D) = 3 microM, clearly establishing Fc gamma RIIa as an IgG4 receptor. 10397151 3 99 theme precise 309:315 arg1 characteristics 317:331 the precise characteristics 305:331 the precise characteristics of IgG binding 305:346 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 5 100 theme IgG 664:666 arg1 interaction 668:678 IgG interaction 664:678 IgG interaction found 664:684 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 5 100 theme IgG 664:666 arg1 IgG3 687:690 IgG3 687:690 IgG3 687:690 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 13 101 theme high 1740:1743 arg1 identity 1756:1763 high amino acid identity 1740:1763 high amino acid identity with Fc gamma RIIa 1740:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 5 102 theme K 718:718 arg1 microM 729:734 K(D) = 0.6 microM 718:734 K(D) = 0.6 microM 718:734 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 13 103 theme acid 1751:1754 arg1 identity 1756:1763 high amino acid identity 1740:1763 high amino acid identity with Fc gamma RIIa 1740:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 12 104 theme Orthorhombic 1450:1461 arg1 crystals 1463:1470 Orthorhombic crystals 1450:1470 Orthorhombic crystals 1450:1470 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 3 105 theme binding 340:346 arg1 characteristics 317:331 the precise characteristics 305:331 the precise characteristics of IgG binding 305:346 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 3 105 theme binding 340:346 arg1 structure 370:378 three-dimensional structure 352:378 three-dimensional structure of Fc gamma RIIa 352:395 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 3 106 theme gamma 386:390 arg1 RIIa 392:395 Fc gamma RIIa 383:395 Fc gamma RIIa 383:395 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 5 107 theme D 720:720 arg1 microM 729:734 K(D) = 0.6 microM 718:734 K(D) = 0.6 microM 718:734 Equilibrium binding analysis of the Fc gamma RII:IgG interaction found, IgG3 binds with an affinity of K(D) = 0.6 microM, as expected. 10397151 13 108 theme gamma 1693:1697 arg1 RIIa 1699:1702 Fc gamma RIIa 1690:1702 Fc gamma RIIa 1690:1702 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 10 109 theme derived 1277:1283 arg1 RIIa 1294:1297 baculovirus derived Fc gamma RIIa 1265:1297 baculovirus derived Fc gamma RIIa 1265:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 12 110 dep dimensions 1552:1561 arg1 A 1572:1572 a = 78.8 A 1563:1572 a = 78.8 A 1563:1572 Orthorhombic crystals diffract X-rays beyond 2.1 A resolution in the space group P2(1)2(1)2 with cell dimensions a = 78.8 A, b = 100.5 A, c = 27.8 A. 10397151 0 111 theme Fc 44:45 arg1 receptor 76:83 the low affinity receptor 59:83 the low affinity receptor for IgG 59:91 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 0 111 theme Fc 44:45 arg1 RIIa 53:56 Fc gamma RIIa 44:56 Fc gamma RIIa 44:56 Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. 10397151 4 112 theme RIIa 471:474 arg1 interaction 480:490 the Fc gamma RIIa:IgG interaction 458:490 the Fc gamma RIIa:IgG interaction 458:490 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 9 113 gly glycosylation 1151:1163 arg2 sites 1165:1169 both potential N-linked glycosylation sites 1127:1169 both potential N-linked glycosylation sites 1127:1169 Furthermore, both potential N-linked glycosylation sites are occupied. 10397151 11 114 theme first 1370:1374 arg1 crystallisation 1376:1390 the first crystallisation 1366:1390 the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa 1366:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 7 115 theme gamma 943:947 arg1 RIIa 949:952 derived Fc gamma RIIa 932:952 derived Fc gamma RIIa 932:952 Biochemical analysis of mammalian and insect cell derived Fc gamma RIIa established the genuine N-terminus with Q being the first amino acid in the sequence Q, A, A, A, P.. 10397151 9 116 link N-linked 1142:1149 arg1 sites 1165:1169 both potential N-linked glycosylation sites 1127:1169 both potential N-linked glycosylation sites 1127:1169 Furthermore, both potential N-linked glycosylation sites are occupied. 10397151 3 117 theme three-dimensional 352:368 arg1 structure 370:378 three-dimensional structure 352:378 three-dimensional structure of Fc gamma RIIa 352:395 However, the precise characteristics of IgG binding and three-dimensional structure of Fc gamma RIIa have not been reported. 10397151 4 118 theme IgG 476:478 arg1 interaction 480:490 the Fc gamma RIIa:IgG interaction 458:490 the Fc gamma RIIa:IgG interaction 458:490 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 13 119 theme proteins 1720:1727 arg1 structure 1677:1685 the three-dimensional structure 1655:1685 the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa 1655:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 10397151 9 120 theme N-linked 1142:1149 arg1 sites 1165:1169 both potential N-linked glycosylation sites 1127:1169 both potential N-linked glycosylation sites 1127:1169 Furthermore, both potential N-linked glycosylation sites are occupied. 10397151 4 121 theme gamma 550:554 arg1 RIIa 556:559 recombinant Fc gamma RIIa 535:559 recombinant Fc gamma RIIa that has been used to generate high quality crystals 535:612 This study describes the affinity of the Fc gamma RIIa:IgG interaction as well as biochemical characterisation of recombinant Fc gamma RIIa that has been used to generate high quality crystals. 10397151 10 122 theme gamma 1288:1292 arg1 RIIa 1294:1297 baculovirus derived Fc gamma RIIa 1265:1297 baculovirus derived Fc gamma RIIa 1265:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 11 123 theme quality 1407:1413 arg1 crystals 1415:1422 diffraction quality crystals 1395:1422 diffraction quality crystals of soluble Fc gamma RIIa 1395:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 10 124 theme side 1332:1335 arg1 chains 1337:1342 core mannose oligosaccharide side chains 1303:1342 core mannose oligosaccharide side chains 1303:1342 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 10 124 theme side 1332:1335 arg1 N-glycans 1252:1260 the N-glycans 1248:1260 the N-glycans of baculovirus derived Fc gamma RIIa 1248:1297 Electrospray ionisation mass spectrometry (ESMS) indicate that the N-glycans of baculovirus derived Fc gamma RIIa are core mannose oligosaccharide side chains. 10397151 11 125 theme Fc 1435:1436 arg1 RIIa 1444:1447 soluble Fc gamma RIIa 1427:1447 soluble Fc gamma RIIa 1427:1447 Finally, we describe the first crystallisation of diffraction quality crystals of soluble Fc gamma RIIa. 10397151 13 126 theme RIIa 1699:1702 arg1 structure 1677:1685 the three-dimensional structure 1655:1685 the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa 1655:1782 This marks a significant advance towards understanding the three-dimensional structure of Fc gamma RIIa and related FcR proteins that share high amino acid identity with Fc gamma RIIa. 21389326 0 0 theme complex 88:94 arg1 assembly 43:50 the extracellular assembly 25:50 the extracellular assembly of the hematopoietic Flt3 signaling complex 25:94 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex. 21389326 1 1 theme signaling 292:300 arg1 cascades 302:309 signaling cascades 292:309 signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells 292:421 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 2 2 theme acute 586:590 arg1 leukemia 600:607 acute myeloid leukemia 586:607 acute myeloid leukemia 586:607 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 4 3 theme extracellular 889:901 arg1 domain 903:908 extracellular domain 3 889:910 extracellular domain 3 of Flt3 889:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 4 3 theme extracellular 889:901 arg1 Flt3 915:918 Flt3 915:918 Flt3 915:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 1 4 theme Fms-like 145:152 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 4 theme Fms-like 145:152 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 7 5 theme FL 1442:1443 arg1 reminiscent 1445:1455 FL reminiscent 1442:1455 FL reminiscent of a "lock-and-key" binding mode 1442:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 0 6 theme signaling 78:86 arg1 complex 88:94 the hematopoietic Flt3 signaling complex 55:94 the hematopoietic Flt3 signaling complex 55:94 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex. 21389326 3 7 theme Flt3 655:658 arg1 complex 676:682 the Flt3 ligand-receptor complex 651:682 the Flt3 ligand-receptor complex 651:682 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 3 8 theme structural 630:639 arg1 basis 641:645 the structural basis 626:645 the structural basis for the Flt3 ligand-receptor complex 626:682 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 1 9 theme tyrosine 154:161 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 9 theme tyrosine 154:161 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 10 theme crucial 311:317 arg1 cascades 302:309 signaling cascades 292:309 signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells 292:421 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 6 11 theme points 1227:1232 arg1 dissection 1195:1204 thermodynamic dissection 1181:1204 thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event 1181:1283 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 7 12 theme preformed 1413:1421 arg1 epitope 1431:1437 a single preformed binding epitope 1404:1437 a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode 1404:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 1 13 theme central 222:228 arg1 roles 230:234 central roles 222:234 central roles 222:234 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 2 14 theme hematologic 494:504 arg1 malignancies 506:517 hematologic malignancies 494:517 hematologic malignancies 494:517 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 3 15 theme unanticipated 698:710 arg1 assembly 726:733 an unanticipated extracellular assembly 695:733 an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date 695:789 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 2 16 theme clinical 566:573 arg1 target 575:580 clinical target 566:580 clinical target for acute myeloid leukemia 566:607 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 7 17 theme binding 1423:1429 arg1 epitope 1431:1437 a single preformed binding epitope 1404:1437 a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode 1404:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 4 18 theme homotypic 964:972 arg1 interactions 983:994 homotypic receptor interactions 964:994 homotypic receptor interactions 964:994 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 4 19 theme interactions 983:994 arg1 devoid 954:959 devoid 954:959 devoid 954:959 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 5 20 with Comparisons 997:1007 arg1 receptors 1033:1041 homologous receptors 1022:1041 homologous receptors 1022:1041 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 3 21 theme extracellular 712:724 arg1 assembly 726:733 an unanticipated extracellular assembly 695:733 an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date 695:789 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 6 22 theme binding 1271:1277 arg1 event 1279:1283 a pronounced enthalpically driven binding event 1237:1283 a pronounced enthalpically driven binding event 1237:1283 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 7 23 theme FL 1372:1373 arg1 complex 1375:1381 the high-affinity Flt3:FL complex 1349:1381 the high-affinity Flt3:FL complex 1349:1381 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 1 24 theme kinase 163:168 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 24 theme kinase 163:168 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 2 25 theme receptors 481:489 arg1 one 446:448 one 446:448 one 446:448 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 2 25 theme receptors 481:489 arg1 receptors 481:489 the most frequently mutated receptors 453:489 the most frequently mutated receptors in hematologic malignancies 453:517 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 6 26 theme driven 1264:1269 arg1 event 1279:1283 a pronounced enthalpically driven binding event 1237:1283 a pronounced enthalpically driven binding event 1237:1283 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 7 27 theme single 1406:1411 arg1 epitope 1431:1437 a single preformed binding epitope 1404:1437 a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode 1404:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 7 28 theme " 1475:1475 arg1 mode 1485:1488 a "lock-and-key" binding mode 1460:1488 a "lock-and-key" binding mode 1460:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 4 29 theme ternary 938:944 arg1 complex 946:952 a ternary complex 936:952 a ternary complex devoid of homotypic receptor interactions 936:994 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 7 30 from epitope 1431:1437 arg1 reminiscent 1445:1455 FL reminiscent 1442:1455 FL reminiscent of a "lock-and-key" binding mode 1442:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 4 31 theme devoid 954:959 arg1 complex 946:952 a ternary complex 936:952 a ternary complex devoid of homotypic receptor interactions 936:994 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 4 32 theme binding 849:855 arg1 epitope 857:863 a remarkably compact binding epitope 828:863 a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3 828:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 2 33 theme mutated 473:479 arg1 receptors 481:489 the most frequently mutated receptors 453:489 the most frequently mutated receptors in hematologic malignancies 453:517 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 6 34 theme pronounced 1239:1248 arg1 event 1279:1283 a pronounced enthalpically driven binding event 1237:1283 a pronounced enthalpically driven binding event 1237:1283 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 0 35 theme Structural 0:9 arg1 insights 11:18 Structural insights 0:18 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex 0:94 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex. 21389326 5 36 theme assembly 1158:1165 arg1 features 1123:1130 the unique features 1112:1130 the unique features of the Flt3 extracellular assembly 1112:1165 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 2 37 theme myeloid 592:598 arg1 leukemia 600:607 acute myeloid leukemia 586:607 acute myeloid leukemia 586:607 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 7 38 theme lock-and-key 1463:1474 arg1 mode 1485:1488 a "lock-and-key" binding mode 1460:1488 a "lock-and-key" binding mode 1460:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 7 39 theme mode 1485:1488 arg1 reminiscent 1445:1455 FL reminiscent 1442:1455 FL reminiscent of a "lock-and-key" binding mode 1442:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 5 40 theme unique 1116:1121 arg1 features 1123:1130 the unique features 1112:1130 the unique features of the Flt3 extracellular assembly 1112:1165 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 3 41 theme other 746:750 arg1 complex 761:767 any other RTKIII/V complex 742:767 any other RTKIII/V complex characterized to date 742:789 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 4 42 theme Flt3 819:822 arg1 dimerization 803:814 dimerization 803:814 dimerization of Flt3 803:822 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 1 43 theme class 101:105 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 43 theme class 101:105 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 5 44 theme available 1047:1055 arg1 data 1069:1072 available mutagenesis data 1047:1072 available mutagenesis data for FL 1047:1079 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 5 45 theme mutagenesis 1057:1067 arg1 data 1069:1072 available mutagenesis data 1047:1072 available mutagenesis data for FL 1047:1079 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 4 46 theme receptor 974:981 arg1 interactions 983:994 homotypic receptor interactions 964:994 homotypic receptor interactions 964:994 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 5 47 with data 1069:1072 arg1 receptors 1033:1041 homologous receptors 1022:1041 homologous receptors 1022:1041 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 0 48 theme extracellular 29:41 arg1 assembly 43:50 the extracellular assembly 25:50 the extracellular assembly of the hematopoietic Flt3 signaling complex 25:94 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex. 21389326 1 49 theme immune 261:266 arg1 system 268:273 the immune system 257:273 the immune system 257:273 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 4 50 theme Flt3 915:918 arg1 domain 903:908 extracellular domain 3 889:910 extracellular domain 3 of Flt3 889:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 4 50 theme Flt3 915:918 arg1 Flt3 915:918 Flt3 915:918 Flt3 915:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 1 51 theme hematopoietic 358:370 arg1 progenitors 372:382 hematopoietic progenitors 358:382 hematopoietic progenitors 358:382 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 5 52 theme Flt3 1139:1142 arg1 assembly 1158:1165 the Flt3 extracellular assembly 1135:1165 the Flt3 extracellular assembly 1135:1165 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 6 53 theme entropic 1299:1306 arg1 penalty 1308:1314 an entropic penalty 1296:1314 an entropic penalty 1296:1314 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 1 54 theme progenitors 372:382 arg1 homeostasis 343:353 homeostasis 343:353 homeostasis 343:353 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 54 theme progenitors 372:382 arg1 development 327:337 the development 323:337 the development 323:337 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 5 55 theme Flt3 1012:1015 arg1 data 1069:1072 available mutagenesis data 1047:1072 available mutagenesis data for FL 1047:1079 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 5 55 theme Flt3 1012:1015 arg1 Comparisons 997:1007 Comparisons 997:1007 Comparisons of Flt3 with homologous receptors 997:1041 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 4 56 theme domain 903:908 arg1 tip 882:884 the tip 878:884 the tip of extracellular domain 3 of Flt3 878:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 1 57 theme receptor 111:118 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 57 theme receptor 111:118 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 3 58 theme ligand-receptor 660:674 arg1 complex 676:682 the Flt3 ligand-receptor complex 651:682 the Flt3 ligand-receptor complex 651:682 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 3 59 theme RTKIII/V 752:759 arg1 complex 761:767 any other RTKIII/V complex 742:767 any other RTKIII/V complex characterized to date 742:789 Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date. 21389326 5 60 theme homologous 1022:1031 arg1 receptors 1033:1041 homologous receptors 1022:1041 homologous receptors 1022:1041 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 2 61 from receptors 481:489 arg1 malignancies 506:517 hematologic malignancies 494:517 hematologic malignancies 494:517 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 1 62 theme tyrosine 120:127 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 62 theme tyrosine 120:127 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 6 63 theme formation 1217:1225 arg1 points 1227:1232 complex formation points 1209:1232 complex formation points 1209:1232 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 5 64 theme extracellular 1144:1156 arg1 assembly 1158:1165 the Flt3 extracellular assembly 1135:1165 the Flt3 extracellular assembly 1135:1165 Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly. 21389326 2 65 theme prognostic 544:553 arg1 factor 555:560 a major prognostic factor 536:560 a major prognostic factor 536:560 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 1 66 theme antigen-presenting 388:405 arg1 cells 417:421 antigen-presenting dendritic cells 388:421 antigen-presenting dendritic cells 388:421 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 6 67 theme complex 1209:1215 arg1 points 1227:1232 complex formation points 1209:1232 complex formation points 1209:1232 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 1 68 theme kinase 129:134 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 68 theme kinase 129:134 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 4 69 located localized 865:873 arg1 tip 882:884 the tip 878:884 the tip of extracellular domain 3 of Flt3 878:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 4 69 located localized 865:873 arg2 epitope 857:863 a remarkably compact binding epitope 828:863 a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3 828:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 1 70 theme cytokine 196:203 arg1 FL 213:214 FL 213:214 FL 213:214 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 70 theme cytokine 196:203 arg1 ligand 205:210 its cytokine ligand 192:210 its cytokine ligand (FL) 192:215 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 2 71 theme major 538:542 arg1 factor 555:560 a major prognostic factor 536:560 a major prognostic factor 536:560 However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia. 21389326 1 72 theme dendritic 407:415 arg1 cells 417:421 antigen-presenting dendritic cells 388:421 antigen-presenting dendritic cells 388:421 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 7 73 theme Flt3 1367:1370 arg1 complex 1375:1381 the high-affinity Flt3:FL complex 1349:1381 the high-affinity Flt3:FL complex 1349:1381 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 0 74 theme Flt3 73:76 arg1 complex 88:94 the hematopoietic Flt3 signaling complex 55:94 the hematopoietic Flt3 signaling complex 55:94 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex. 21389326 1 75 theme cells 417:421 arg1 homeostasis 343:353 homeostasis 343:353 homeostasis 343:353 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 75 theme cells 417:421 arg1 development 327:337 the development 323:337 the development 323:337 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 7 76 theme antagonist 1521:1530 arg1 design 1532:1537 antagonist design 1521:1537 antagonist design 1521:1537 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 1 77 theme RTKIII 137:142 arg1 receptor 170:177 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 97:179 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) 97:186 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 1 77 theme RTKIII 137:142 arg1 Flt3 182:185 Flt3 182:185 Flt3 182:185 The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells. 21389326 0 78 theme hematopoietic 59:71 arg1 complex 88:94 the hematopoietic Flt3 signaling complex 55:94 the hematopoietic Flt3 signaling complex 55:94 Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex. 21389326 7 79 theme high-affinity 1353:1365 arg1 complex 1375:1381 the high-affinity Flt3:FL complex 1349:1381 the high-affinity Flt3:FL complex 1349:1381 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 21389326 6 80 theme thermodynamic 1181:1193 arg1 dissection 1195:1204 thermodynamic dissection 1181:1204 thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event 1181:1283 Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty. 21389326 4 81 theme compact 841:847 arg1 epitope 857:863 a remarkably compact binding epitope 828:863 a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3 828:918 FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions. 21389326 7 82 theme binding 1477:1483 arg1 mode 1485:1488 a "lock-and-key" binding mode 1460:1488 a "lock-and-key" binding mode 1460:1488 Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design. 32616672 0 0 from function 17:24 arg1 infections 73:82 urinary tract infections 59:82 urinary tract infections 59:82 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 5 1 theme tract 950:954 arg1 infections 956:965 urinary tract infections 942:965 urinary tract infections 942:965 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 1 2 theme filament 223:230 arg1 structure 232:240 filament structure 223:240 filament structure 223:240 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 3 3 theme multivalent 517:527 arg1 ligand 529:534 a multivalent ligand 515:534 a multivalent ligand for the bacterial type 1 pilus adhesin 515:573 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 3 3 theme multivalent 517:527 arg1 uromodulin 496:505 uromodulin 496:505 uromodulin 496:505 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 3 4 theme biophysical 463:473 arg1 assays 475:480 biophysical assays 463:480 biophysical assays 463:480 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 0 5 from Architecture 0:11 arg1 infections 73:82 urinary tract infections 59:82 urinary tract infections 59:82 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 3 6 theme type 554:557 arg1 adhesin 567:573 the bacterial type 1 pilus adhesin 540:573 the bacterial type 1 pilus adhesin 540:573 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 2 7 theme cryo-electron 304:316 arg1 tomography 318:327 cryo-electron tomography 304:327 cryo-electron tomography 304:327 We used cryo-electron tomography to show that the uromodulin filament consists of a zigzag-shaped backbone with laterally protruding arms. 32616672 2 8 with backbone 394:401 arg1 arms 429:432 laterally protruding arms 408:432 laterally protruding arms 408:432 We used cryo-electron tomography to show that the uromodulin filament consists of a zigzag-shaped backbone with laterally protruding arms. 32616672 2 9 theme zigzag-shaped 380:392 arg1 backbone 394:401 a zigzag-shaped backbone 378:401 a zigzag-shaped backbone with laterally protruding arms 378:432 We used cryo-electron tomography to show that the uromodulin filament consists of a zigzag-shaped backbone with laterally protruding arms. 32616672 3 10 theme bacterial 544:552 arg1 adhesin 567:573 the bacterial type 1 pilus adhesin 540:573 the bacterial type 1 pilus adhesin 540:573 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 5 11 theme enigmatic 983:991 arg1 roles 993:997 its more enigmatic roles 974:997 its more enigmatic roles in physiology and disease 974:1023 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 1 12 theme protection 259:268 arg1 structure 232:240 filament structure 223:240 filament structure 223:240 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 1 12 theme protection 259:268 arg1 mechanism 246:254 mechanism 246:254 mechanism 246:254 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 1 13 from protein 117:123 arg1 urine 134:138 human urine 128:138 human urine 128:138 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 3 14 theme pilus 561:565 arg1 adhesin 567:573 the bacterial type 1 pilus adhesin 540:573 the bacterial type 1 pilus adhesin 540:573 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 3 15 from epitopes 596:603 arg1 arms 629:632 the regularly spaced arms 608:632 the regularly spaced arms 608:632 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 2 16 theme protruding 418:427 arg1 arms 429:432 laterally protruding arms 408:432 laterally protruding arms 408:432 We used cryo-electron tomography to show that the uromodulin filament consists of a zigzag-shaped backbone with laterally protruding arms. 32616672 0 17 theme uromodulin 35:44 arg1 filaments 46:54 human uromodulin filaments 29:54 human uromodulin filaments 29:54 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 4 18 theme uromodulin-uropathogen 646:667 arg1 interactions 669:680 uromodulin-uropathogen interactions 646:680 uromodulin-uropathogen interactions 646:680 Imaging of uromodulin-uropathogen interactions in vitro and in patient urine showed that uromodulin filaments associate with uropathogens and mediate bacterial aggregation, which likely prevents adhesion and allows clearance by micturition. 32616672 5 19 from roles 993:997 arg1 disease 1017:1023 disease 1017:1023 disease 1017:1023 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 5 19 from roles 993:997 arg1 physiology 1002:1011 physiology 1002:1011 physiology 1002:1011 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 4 20 theme interactions 669:680 arg1 Imaging 635:641 Imaging 635:641 Imaging of uromodulin-uropathogen interactions in vitro and in patient urine 635:710 Imaging of uromodulin-uropathogen interactions in vitro and in patient urine showed that uromodulin filaments associate with uropathogens and mediate bacterial aggregation, which likely prevents adhesion and allows clearance by micturition. 32616672 0 21 theme human 29:33 arg1 filaments 46:54 human uromodulin filaments 29:54 human uromodulin filaments 29:54 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 1 22 theme uropathogens 196:207 arg1 adhesion 184:191 the adhesion 180:191 the adhesion of uropathogens 180:207 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 3 23 theme N-glycosylation 435:449 arg1 mapping 451:457 N-glycosylation mapping 435:457 N-glycosylation mapping 435:457 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 3 24 theme specific 587:594 arg1 epitopes 596:603 specific epitopes 587:603 specific epitopes on the regularly spaced arms 587:632 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 1 25 theme abundant 108:115 arg1 protein 117:123 the most abundant protein 99:123 the most abundant protein in human urine 99:138 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 1 25 theme abundant 108:115 arg1 Uromodulin 85:94 Uromodulin 85:94 Uromodulin 85:94 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 0 26 theme filaments 46:54 arg1 function 17:24 function 17:24 function 17:24 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 0 26 theme filaments 46:54 arg1 Architecture 0:11 Architecture 0:11 Architecture 0:11 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 4 27 theme uromodulin 724:733 arg1 filaments 735:743 uromodulin filaments 724:743 uromodulin filaments 724:743 Imaging of uromodulin-uropathogen interactions in vitro and in patient urine showed that uromodulin filaments associate with uropathogens and mediate bacterial aggregation, which likely prevents adhesion and allows clearance by micturition. 32616672 4 28 theme patient 698:704 arg1 urine 706:710 patient urine 698:710 patient urine 698:710 Imaging of uromodulin-uropathogen interactions in vitro and in patient urine showed that uromodulin filaments associate with uropathogens and mediate bacterial aggregation, which likely prevents adhesion and allows clearance by micturition. 32616672 4 29 theme bacterial 785:793 arg1 aggregation 795:805 bacterial aggregation 785:805 bacterial aggregation 785:805 Imaging of uromodulin-uropathogen interactions in vitro and in patient urine showed that uromodulin filaments associate with uropathogens and mediate bacterial aggregation, which likely prevents adhesion and allows clearance by micturition. 32616672 1 30 theme human 128:132 arg1 urine 134:138 human urine 128:138 human urine 128:138 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 0 31 theme tract 67:71 arg1 infections 73:82 urinary tract infections 59:82 urinary tract infections 59:82 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 1 32 dep structure 232:240 arg1 the 219:221 the 219:221 the 219:221 Uromodulin is the most abundant protein in human urine, and it forms filaments that antagonize the adhesion of uropathogens; however, the filament structure and mechanism of protection remain poorly understood. 32616672 5 33 from uromodulin 928:937 arg1 infections 956:965 urinary tract infections 942:965 urinary tract infections 942:965 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 5 33 from uromodulin 928:937 arg1 roles 993:997 its more enigmatic roles 974:997 its more enigmatic roles in physiology and disease 974:1023 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 5 34 theme urinary 942:948 arg1 infections 956:965 urinary tract infections 942:965 urinary tract infections 942:965 These results provide a framework for understanding uromodulin in urinary tract infections and in its more enigmatic roles in physiology and disease. 32616672 2 35 used used 299:302 arg2 We 296:297 We 296:297 We 296:297 We used cryo-electron tomography to show that the uromodulin filament consists of a zigzag-shaped backbone with laterally protruding arms. 32616672 0 36 theme urinary 59:65 arg1 infections 73:82 urinary tract infections 59:82 urinary tract infections 59:82 Architecture and function of human uromodulin filaments in urinary tract infections. 32616672 3 37 theme spaced 622:627 arg1 arms 629:632 the regularly spaced arms 608:632 the regularly spaced arms 608:632 N-glycosylation mapping and biophysical assays revealed that uromodulin acts as a multivalent ligand for the bacterial type 1 pilus adhesin, presenting specific epitopes on the regularly spaced arms. 32616672 2 38 theme uromodulin 346:355 arg1 filament 357:364 the uromodulin filament 342:364 the uromodulin filament 342:364 We used cryo-electron tomography to show that the uromodulin filament consists of a zigzag-shaped backbone with laterally protruding arms. 33196145 1 0 theme domain 322:327 arg1 " 328:328 domain" 322:328 zona pellucida (ZP) "domain" 301:328 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 5 1 theme coat 1093:1096 arg1 filaments 1098:1106 heteromeric vertebrate egg coat filaments 1066:1106 heteromeric vertebrate egg coat filaments 1066:1106 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 0 2 theme module 63:68 arg1 polymer 70:76 a zona pellucida module polymer 46:76 a zona pellucida module polymer 46:76 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 3 3 theme ZP 641:642 arg1 molecule 662:669 an archetypal ZP module-containing molecule 627:669 an archetypal ZP module-containing molecule 627:669 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 4 theme module-containing 644:660 arg1 molecule 662:669 an archetypal ZP module-containing molecule 627:669 an archetypal ZP module-containing molecule 627:669 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 4 5 theme one-start 720:728 arg1 helix 730:734 a one-start helix 718:734 a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation 718:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 1 6 theme ubiquitous 259:268 arg1 module 285:290 a ubiquitous polymerization module 257:290 a ubiquitous polymerization module known as zona pellucida (ZP) "domain" 257:328 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 3 7 from protein 600:606 arg1 urine 617:621 human urine 611:621 human urine 611:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 8 theme mature 679:684 arg1 state 700:704 its mature homopolymeric state 675:704 its mature homopolymeric state 675:704 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 9 theme cryo-electron 507:519 arg1 microscopy 521:530 cryo-electron microscopy 507:530 a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine 505:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 1 10 theme polymerization 270:283 arg1 module 285:290 a ubiquitous polymerization module 257:290 a ubiquitous polymerization module known as zona pellucida (ZP) "domain" 257:328 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 3 11 theme homopolymeric 686:698 arg1 state 700:704 its mature homopolymeric state 675:704 its mature homopolymeric state 675:704 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 2 12 theme protein 480:486 arg1 conformation 450:461 the filamentous conformation 434:461 the filamentous conformation of any ZP module protein 434:486 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 1 13 dep pellucida 306:314 arg1 " 328:328 domain" 322:328 zona pellucida (ZP) "domain" 301:328 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 4 14 theme interdomain 804:814 arg1 linkers 816:822 interdomain linkers 804:822 interdomain linkers that have completely reorganized as a result of propeptide dissociation 804:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 2 15 theme module 473:478 arg1 protein 480:486 any ZP module protein 466:486 any ZP module protein 466:486 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 0 16 dep polymer 70:76 arg1 structure 8:16 Cryo-EM structure 0:16 Cryo-EM structure of native human uromodulin 0:43 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 5 17 theme filaments 1098:1106 arg1 models 1056:1061 UMOD-based models 1045:1061 UMOD-based models of heteromeric vertebrate egg coat filaments 1045:1106 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 2 18 theme element 364:370 arg1 conservation 343:354 the conservation 339:354 the conservation of this element from hydra to humans 339:391 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 0 19 theme Cryo-EM 0:6 arg1 structure 8:16 Cryo-EM structure 0:16 Cryo-EM structure of native human uromodulin 0:43 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 3 20 theme uromodulin 541:550 arg1 protein 600:606 the most abundant protein 582:606 the most abundant protein in human urine 582:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 20 theme uromodulin 541:550 arg1 protein 573:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 5 21 theme heteromeric 1066:1076 arg1 filaments 1098:1106 heteromeric vertebrate egg coat filaments 1066:1106 heteromeric vertebrate egg coat filaments 1066:1106 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 0 22 theme native 21:26 arg1 uromodulin 34:43 native human uromodulin 21:43 native human uromodulin 21:43 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 4 23 with helix 730:734 arg1 twist 769:773 an unprecedented 180-degree twist 741:773 an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation 741:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 5 24 theme binding 992:998 arg1 sites 1000:1004 binding sites 992:1004 binding sites 992:1004 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 2 25 from conservation 343:354 arg1 hydra 377:381 hydra 377:381 hydra 377:381 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 3 26 theme microscopy 521:530 arg1 study 532:536 a cryo-electron microscopy study 505:536 a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine 505:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 2 27 theme detailed 397:404 arg1 information 406:416 no detailed information 394:416 no detailed information 394:416 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 5 28 theme sperm-binding 1126:1138 arg1 region 1140:1145 a common sperm-binding region 1117:1145 a common sperm-binding region 1117:1145 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 1 29 theme extracellular 91:103 arg1 filaments 105:113 extracellular filaments 91:113 extracellular filaments 91:113 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 0 30 theme uromodulin 34:43 arg1 structure 8:16 Cryo-EM structure 0:16 Cryo-EM structure of native human uromodulin 0:43 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 1 31 theme zona 301:304 arg1 ZP 317:318 ZP 317:318 ZP 317:318 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 31 theme zona 301:304 arg1 pellucida 306:314 zona pellucida 301:314 zona pellucida (ZP) "domain" 301:328 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 3 32 theme human 611:615 arg1 urine 617:621 human urine 611:621 human urine 611:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 0 33 theme human 28:32 arg1 uromodulin 34:43 native human uromodulin 21:43 native human uromodulin 21:43 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 2 34 theme filamentous 438:448 arg1 conformation 450:461 the filamentous conformation 434:461 the filamentous conformation of any ZP module protein 434:486 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 3 35 theme abundant 591:598 arg1 protein 600:606 the most abundant protein 582:606 the most abundant protein in human urine 582:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 35 theme abundant 591:598 arg1 protein 573:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 36 theme /Tamm-Horsfall 558:571 arg1 protein 600:606 the most abundant protein 582:606 the most abundant protein in human urine 582:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 36 theme /Tamm-Horsfall 558:571 arg1 protein 573:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 37 from molecule 662:669 arg1 state 700:704 its mature homopolymeric state 675:704 its mature homopolymeric state 675:704 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 5 38 theme egg 1089:1091 arg1 filaments 1098:1106 heteromeric vertebrate egg coat filaments 1066:1106 heteromeric vertebrate egg coat filaments 1066:1106 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 5 39 theme uropathogenic 1017:1029 arg1 bacteria 1031:1038 uropathogenic bacteria 1017:1038 uropathogenic bacteria 1017:1038 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 1 40 theme filaments 105:113 arg1 Assembly 79:86 Assembly 79:86 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense 79:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 3 41 theme UMOD 553:556 arg1 protein 600:606 the most abundant protein 582:606 the most abundant protein in human urine 582:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 41 theme UMOD 553:556 arg1 protein 573:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 uromodulin (UMOD)/Tamm-Horsfall protein 541:579 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 2 42 from hydra 377:381 arg1 conservation 343:354 the conservation 339:354 the conservation of this element from hydra to humans 339:391 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 2 42 from hydra 377:381 arg1 element 364:370 this element 359:370 this element from hydra 359:381 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 5 43 theme vertebrate 1078:1087 arg1 filaments 1098:1106 heteromeric vertebrate egg coat filaments 1066:1106 heteromeric vertebrate egg coat filaments 1066:1106 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 5 44 theme sites 1000:1004 arg1 checkerboard 976:987 a checkerboard 974:987 a checkerboard of binding sites to capture uropathogenic bacteria 974:1038 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 4 45 theme dissociation 883:894 arg1 result 862:867 a result 860:867 a result of propeptide dissociation 860:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 3 46 theme archetypal 630:639 arg1 molecule 662:669 an archetypal ZP module-containing molecule 627:669 an archetypal ZP module-containing molecule 627:669 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 4 47 theme unprecedented 744:756 arg1 twist 769:773 an unprecedented 180-degree twist 741:773 an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation 741:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 1 48 theme matrices 119:126 arg1 Assembly 79:86 Assembly 79:86 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense 79:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 3 49 from study 532:536 arg1 state 700:704 its mature homopolymeric state 675:704 its mature homopolymeric state 675:704 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 1 50 theme antibacterial 222:234 arg1 defense 236:242 antibacterial defense 222:242 antibacterial defense 222:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 4 51 theme propeptide 872:881 arg1 dissociation 883:894 propeptide dissociation 872:894 propeptide dissociation 872:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 5 52 theme UMOD-based 1045:1054 arg1 models 1056:1061 UMOD-based models 1045:1061 UMOD-based models of heteromeric vertebrate egg coat filaments 1045:1106 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 5 53 theme Lateral 897:903 arg1 interaction 905:915 Lateral interaction 897:915 Lateral interaction between filaments in the urine 897:946 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 5 54 from filaments 925:933 arg1 urine 942:946 the urine 938:946 the urine 938:946 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 0 55 theme pellucida 53:61 arg1 polymer 70:76 a zona pellucida module polymer 46:76 a zona pellucida module polymer 46:76 Cryo-EM structure of native human uromodulin, a zona pellucida module polymer. 33196145 3 56 theme protein 573:579 arg1 molecule 662:669 an archetypal ZP module-containing molecule 627:669 an archetypal ZP module-containing molecule 627:669 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 3 56 theme protein 573:579 arg1 study 532:536 a cryo-electron microscopy study 505:536 a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine 505:621 Here, we report a cryo-electron microscopy study of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. 33196145 1 57 theme fundamental 138:148 arg1 fertilization 203:215 fertilization 203:215 fertilization 203:215 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 57 theme fundamental 138:148 arg1 hearing 194:200 hearing 194:200 hearing 194:200 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 57 theme fundamental 138:148 arg1 processes 161:169 fundamental biological processes 138:169 fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense 138:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 57 theme fundamental 138:148 arg1 defense 236:242 antibacterial defense 222:242 antibacterial defense 222:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 57 theme fundamental 138:148 arg1 morphogenesis 179:191 morphogenesis 179:191 morphogenesis 179:191 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 2 58 theme ZP 470:471 arg1 protein 480:486 any ZP module protein 466:486 any ZP module protein 466:486 Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. 33196145 5 59 theme common 1119:1124 arg1 region 1140:1145 a common sperm-binding region 1117:1145 a common sperm-binding region 1117:1145 Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of heteromeric vertebrate egg coat filaments identify a common sperm-binding region at the interface between subunits. 33196145 4 60 theme 180-degree 758:767 arg1 twist 769:773 an unprecedented 180-degree twist 741:773 an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation 741:894 UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. 33196145 1 61 theme biological 150:159 arg1 fertilization 203:215 fertilization 203:215 fertilization 203:215 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 61 theme biological 150:159 arg1 hearing 194:200 hearing 194:200 hearing 194:200 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 61 theme biological 150:159 arg1 processes 161:169 fundamental biological processes 138:169 fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense 138:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 61 theme biological 150:159 arg1 defense 236:242 antibacterial defense 222:242 antibacterial defense 222:242 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 33196145 1 61 theme biological 150:159 arg1 morphogenesis 179:191 morphogenesis 179:191 morphogenesis 179:191 Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) "domain". 35273390 0 0 theme uromodulin 58:67 arg1 module 23:28 the decoy module 13:28 the decoy module of human glycoprotein 2 and uromodulin 13:67 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 2 1 theme X-ray 330:334 arg1 crystallography 336:350 X-ray crystallography 330:350 X-ray crystallography 330:350 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 0 2 theme bacterial 94:102 arg1 FimH 112:115 bacterial adhesin FimH 94:115 bacterial adhesin FimH 94:115 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 2 3 theme new 432:434 arg1 fold 436:439 fold 436:439 fold 436:439 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 3 4 theme UMOD 521:524 arg1 mutations 526:534 UMOD mutations 521:534 UMOD mutations associated with kidney diseases 521:566 The structure rationalizes UMOD mutations associated with kidney diseases and visualizes a key epitope implicated in cast nephropathy. 35273390 1 5 theme uromodulin 143:152 arg1 filaments 161:169 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments 118:169 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments 118:169 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 2 6 theme high-mannose 454:465 arg1 glycan 467:472 the high-mannose glycan 450:472 the high-mannose glycan recognized by FimH 450:491 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 0 7 with interaction 77:87 arg1 FimH 112:115 bacterial adhesin FimH 94:115 bacterial adhesin FimH 94:115 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 0 8 theme decoy 17:21 arg1 module 23:28 the decoy module 13:28 the decoy module of human glycoprotein 2 and uromodulin 13:67 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 2 9 theme bipartite 403:411 arg1 module 419:424 a bipartite decoy module 401:424 a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH 401:491 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 1 10 theme bacterial 257:265 arg1 FimH 283:286 bacterial fimbrial lectin FimH 257:286 bacterial fimbrial lectin FimH 257:286 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 0 11 theme adhesin 104:110 arg1 FimH 112:115 bacterial adhesin FimH 94:115 bacterial adhesin FimH 94:115 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 3 12 theme kidney 552:557 arg1 diseases 559:566 kidney diseases 552:566 kidney diseases 552:566 The structure rationalizes UMOD mutations associated with kidney diseases and visualizes a key epitope implicated in cast nephropathy. 35273390 1 13 theme fimbrial 267:274 arg1 FimH 283:286 bacterial fimbrial lectin FimH 257:286 bacterial fimbrial lectin FimH 257:286 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 3 14 theme key 585:587 arg1 epitope 589:595 a key epitope 583:595 a key epitope implicated in cast nephropathy 583:626 The structure rationalizes UMOD mutations associated with kidney diseases and visualizes a key epitope implicated in cast nephropathy. 35273390 0 15 theme module 23:28 arg1 interaction 77:87 its interaction 73:87 its interaction with bacterial adhesin FimH 73:115 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 0 15 theme module 23:28 arg1 Structure 0:8 Structure 0:8 Structure of the decoy module of human glycoprotein 2 and uromodulin 0:67 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 1 16 theme lectin 276:281 arg1 FimH 283:286 bacterial fimbrial lectin FimH 257:286 bacterial fimbrial lectin FimH 257:286 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 2 17 theme decoy 413:417 arg1 module 419:424 a bipartite decoy module 401:424 a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH 401:491 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 0 18 theme glycoprotein 39:50 arg1 module 23:28 the decoy module 13:28 the decoy module of human glycoprotein 2 and uromodulin 13:67 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 0 19 theme human 33:37 arg1 glycoprotein 39:50 human glycoprotein 2 33:52 human glycoprotein 2 33:52 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 3 20 theme cast 611:614 arg1 nephropathy 616:626 cast nephropathy 611:626 cast nephropathy 611:626 The structure rationalizes UMOD mutations associated with kidney diseases and visualizes a key epitope implicated in cast nephropathy. 35273390 0 21 with Structure 0:8 arg1 FimH 112:115 bacterial adhesin FimH 94:115 bacterial adhesin FimH 94:115 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 2 22 theme AlphaFold2 302:311 arg1 predictions 313:323 AlphaFold2 predictions 302:323 AlphaFold2 predictions 302:323 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 1 23 theme gastrointestinal 187:202 arg1 infections 222:231 gastrointestinal and urinary tract infections 187:231 infections 222:231 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 2 24 contain contain 393:399 arg1 proteins 384:391 these proteins 378:391 these proteins 378:391 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 2 24 contain contain 393:399 arg2 module 419:424 a bipartite decoy module 401:424 a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH 401:491 By combining AlphaFold2 predictions with X-ray crystallography and cryo-EM, we show that these proteins contain a bipartite decoy module whose new fold presents the high-mannose glycan recognized by FimH. 35273390 1 25 theme Glycoprotein 118:129 arg1 filaments 161:169 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments 118:169 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments 118:169 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 1 26 theme urinary 208:214 arg1 infections 222:231 gastrointestinal and urinary tract infections 187:231 infections 222:231 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 35273390 0 27 gly glycoprotein 39:50 arg1 glycoprotein 39:50 human glycoprotein 2 33:52 human glycoprotein 2 33:52 Structure of the decoy module of human glycoprotein 2 and uromodulin and its interaction with bacterial adhesin FimH. 35273390 1 28 theme tract 216:220 arg1 infections 222:231 gastrointestinal and urinary tract infections 187:231 infections 222:231 Glycoprotein 2 (GP2) and uromodulin (UMOD) filaments protect against gastrointestinal and urinary tract infections by acting as decoys for bacterial fimbrial lectin FimH. 19053759 4 0 theme ureido 579:584 arg1 linkage 586:592 The ureido linkage 575:592 The ureido linkage between P1 and P1' inhibitor sites 575:627 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 6 1 theme S1 975:976 arg1 site 978:981 the S1 site 971:981 the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity 971:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 5 2 theme inhibitors 843:852 arg1 chains 867:872 the side chains 858:872 the side chains of Arg534, Arg536, and Asn519 858:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 2 theme inhibitors 843:852 arg1 Asn519 897:902 Asn519 897:902 Asn519 897:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 2 theme inhibitors 843:852 arg1 inhibitors 843:852 the inhibitors 839:852 the inhibitors 839:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 2 theme inhibitors 843:852 arg1 Arg534 877:882 Arg534 877:882 Arg534 877:882 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 2 theme inhibitors 843:852 arg1 Arg536 885:890 Arg536 885:890 Arg536 885:890 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 2 theme inhibitors 843:852 arg1 group 830:834 the P1 carboxylate group 811:834 the P1 carboxylate group of the inhibitors 811:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 0 3 theme structural 84:93 arg1 characterization 95:110 structural characterization 84:110 structural characterization 84:110 Interactions between human glutamate carboxypeptidase II and urea-based inhibitors: structural characterization. 19053759 6 4 theme inhibitors 1047:1056 arg1 design 1025:1030 structure-based design 1009:1030 structure-based design of novel GCPII inhibitors with increased lipophilicity 1009:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 6 5 theme structure-based 1009:1023 arg1 design 1025:1030 structure-based design 1009:1030 structure-based design of novel GCPII inhibitors with increased lipophilicity 1009:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 6 6 with inhibitors 1047:1056 arg1 lipophilicity 1073:1085 increased lipophilicity 1063:1085 increased lipophilicity 1063:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 6 7 theme increased 1063:1071 arg1 lipophilicity 1073:1085 increased lipophilicity 1063:1085 increased lipophilicity 1063:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 2 8 theme further 330:336 arg1 development 338:348 further development 330:348 further development of such compounds 330:366 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 5 9 theme Asn519 897:902 arg1 chains 867:872 the side chains 858:872 the side chains of Arg534, Arg536, and Asn519 858:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 9 theme Asn519 897:902 arg1 Asn519 897:902 Asn519 897:902 Asn519 897:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 9 theme Asn519 897:902 arg1 inhibitors 843:852 the inhibitors 839:852 the inhibitors 839:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 9 theme Asn519 897:902 arg1 Arg534 877:882 Arg534 877:882 Arg534 877:882 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 9 theme Asn519 897:902 arg1 Arg536 885:890 Arg536 885:890 Arg536 885:890 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 9 theme Asn519 897:902 arg1 group 830:834 the P1 carboxylate group 811:834 the P1 carboxylate group of the inhibitors 811:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 1 10 theme imaging 283:289 arg1 ligands 146:152 Urea-based, low molecular weight ligands 113:152 ligands 146:152 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 10 theme imaging 283:289 arg1 agents 291:296 imaging agents 283:296 imaging agents for prostate cancer 283:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 5 11 theme bonds 797:801 arg1 network 777:783 a network 775:783 a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519 775:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 2 12 theme compounds 358:366 arg1 development 338:348 further development 330:348 further development of such compounds 330:366 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 5 13 theme hydrogen 788:795 arg1 bonds 797:801 hydrogen bonds 788:801 hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519 788:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 4 14 theme Tyr552 697:702 arg1 ion 670:672 the active-site Zn(1)(2+) ion 644:672 the active-site Zn(1)(2+) ion 644:672 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 4 14 theme Tyr552 697:702 arg1 chains 687:692 the side chains 678:692 the side chains of Tyr552 and His553 678:713 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 5 15 theme carboxylate 818:828 arg1 inhibitors 843:852 the inhibitors 839:852 the inhibitors 839:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 15 theme carboxylate 818:828 arg1 Asn519 897:902 Asn519 897:902 Asn519 897:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 15 theme carboxylate 818:828 arg1 Arg534 877:882 Arg534 877:882 Arg534 877:882 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 15 theme carboxylate 818:828 arg1 Arg536 885:890 Arg536 885:890 Arg536 885:890 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 15 theme carboxylate 818:828 arg1 group 830:834 the P1 carboxylate group 811:834 the P1 carboxylate group of the inhibitors 811:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 2 16 theme complexes 408:416 arg1 structures 389:398 X-ray structures 383:398 X-ray structures of four complexes between human GCPII and urea-based inhibitors 383:462 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 5 17 theme Arg534 877:882 arg1 chains 867:872 the side chains 858:872 the side chains of Arg534, Arg536, and Asn519 858:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 17 theme Arg534 877:882 arg1 Asn519 897:902 Asn519 897:902 Asn519 897:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 17 theme Arg534 877:882 arg1 inhibitors 843:852 the inhibitors 839:852 the inhibitors 839:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 17 theme Arg534 877:882 arg1 Arg534 877:882 Arg534 877:882 Arg534 877:882 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 17 theme Arg534 877:882 arg1 Arg536 885:890 Arg536 885:890 Arg536 885:890 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 17 theme Arg534 877:882 arg1 group 830:834 the P1 carboxylate group 811:834 the P1 carboxylate group of the inhibitors 811:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 3 18 theme invariant 511:519 arg1 moiety 531:536 an invariant glutarate moiety 508:536 an invariant glutarate moiety within the S1' pocket of the enzyme 508:572 All ligands demonstrate an invariant glutarate moiety within the S1' pocket of the enzyme. 19053759 4 19 theme Zn 660:661 arg1 ion 670:672 the active-site Zn(1)(2+) ion 644:672 the active-site Zn(1)(2+) ion 644:672 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 2 20 theme high 467:470 arg1 resolution 472:481 high resolution 467:481 high resolution 467:481 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 0 21 dep characterization 95:110 arg1 Interactions 0:11 Interactions 0:11 Interactions between human glutamate carboxypeptidase II and urea-based inhibitors 0:81 Interactions between human glutamate carboxypeptidase II and urea-based inhibitors: structural characterization. 19053759 4 22 theme active-site 648:658 arg1 ion 670:672 the active-site Zn(1)(2+) ion 644:672 the active-site Zn(1)(2+) ion 644:672 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 1 23 theme Urea-based 113:122 arg1 ligands 146:152 Urea-based, low molecular weight ligands 113:152 ligands 146:152 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 23 theme Urea-based 113:122 arg1 agents 291:296 imaging agents 283:296 imaging agents for prostate cancer 283:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 0 24 theme glutamate 27:35 arg1 carboxypeptidase 37:52 human glutamate carboxypeptidase II 21:55 human glutamate carboxypeptidase II 21:55 Interactions between human glutamate carboxypeptidase II and urea-based inhibitors: structural characterization. 19053759 3 25 theme enzyme 567:572 arg1 pocket 553:558 the S1' pocket 545:558 the S1' pocket of the enzyme 545:572 All ligands demonstrate an invariant glutarate moiety within the S1' pocket of the enzyme. 19053759 2 26 theme urea-based 442:451 arg1 inhibitors 453:462 urea-based inhibitors 442:462 urea-based inhibitors 442:462 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 1 27 theme prostate 302:309 arg1 cancer 311:316 prostate cancer 302:316 prostate cancer 302:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 6 28 theme hydrophobic 939:949 arg1 pocket 951:956 a hydrophobic pocket 937:956 a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity 937:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 1 29 theme low 125:127 arg1 ligands 146:152 Urea-based, low molecular weight ligands 113:152 ligands 146:152 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 29 theme low 125:127 arg1 agents 291:296 imaging agents 283:296 imaging agents for prostate cancer 283:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 30 theme glutamate 157:165 arg1 carboxypeptidase 167:182 glutamate carboxypeptidase II 157:185 glutamate carboxypeptidase II (GCPII) 157:193 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 30 theme glutamate 157:165 arg1 GCPII 188:192 GCPII 188:192 GCPII 188:192 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 5 31 theme S1 740:741 arg1 pocket 743:748 the S1 pocket 736:748 the S1 pocket 736:748 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 2 32 theme such 353:356 arg1 compounds 358:366 such compounds 353:366 such compounds 353:366 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 1 33 theme various 225:231 arg1 models 233:238 various models 225:238 various models of neurological disorders 225:264 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 2 34 theme human 426:430 arg1 GCPII 432:436 human GCPII 426:436 human GCPII 426:436 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 4 35 theme 2+ 666:667 arg1 ion 670:672 the active-site Zn(1)(2+) ion 644:672 the active-site Zn(1)(2+) ion 644:672 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 5 36 theme side 862:865 arg1 chains 867:872 the side chains 858:872 the side chains of Arg534, Arg536, and Asn519 858:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 0 37 theme urea-based 61:70 arg1 inhibitors 72:81 urea-based inhibitors 61:81 urea-based inhibitors 61:81 Interactions between human glutamate carboxypeptidase II and urea-based inhibitors: structural characterization. 19053759 1 38 theme molecular 129:137 arg1 ligands 146:152 Urea-based, low molecular weight ligands 113:152 ligands 146:152 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 38 theme molecular 129:137 arg1 agents 291:296 imaging agents 283:296 imaging agents for prostate cancer 283:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 5 39 theme Arg536 885:890 arg1 chains 867:872 the side chains 858:872 the side chains of Arg534, Arg536, and Asn519 858:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 39 theme Arg536 885:890 arg1 Asn519 897:902 Asn519 897:902 Asn519 897:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 39 theme Arg536 885:890 arg1 inhibitors 843:852 the inhibitors 839:852 the inhibitors 839:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 39 theme Arg536 885:890 arg1 Arg534 877:882 Arg534 877:882 Arg534 877:882 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 39 theme Arg536 885:890 arg1 Arg536 885:890 Arg536 885:890 Arg536 885:890 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 39 theme Arg536 885:890 arg1 group 830:834 the P1 carboxylate group 811:834 the P1 carboxylate group of the inhibitors 811:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 1 40 from efficacy 213:220 arg1 models 233:238 various models 225:238 various models of neurological disorders 225:264 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 41 theme weight 139:144 arg1 ligands 146:152 Urea-based, low molecular weight ligands 113:152 ligands 146:152 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 41 theme weight 139:144 arg1 agents 291:296 imaging agents 283:296 imaging agents for prostate cancer 283:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 6 42 theme GCPII 1041:1045 arg1 inhibitors 1047:1056 novel GCPII inhibitors 1035:1056 novel GCPII inhibitors with increased lipophilicity 1035:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 4 43 theme His553 708:713 arg1 ion 670:672 the active-site Zn(1)(2+) ion 644:672 the active-site Zn(1)(2+) ion 644:672 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 4 43 theme His553 708:713 arg1 chains 687:692 the side chains 678:692 the side chains of Tyr552 and His553 678:713 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 6 44 theme accessory 958:966 arg1 pocket 951:956 a hydrophobic pocket 937:956 a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity 937:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 6 45 theme novel 1035:1039 arg1 inhibitors 1047:1056 novel GCPII inhibitors 1035:1056 novel GCPII inhibitors with increased lipophilicity 1035:1085 Importantly, we have identified a hydrophobic pocket accessory to the S1 site that can be exploited for structure-based design of novel GCPII inhibitors with increased lipophilicity. 19053759 4 46 theme side 682:685 arg1 chains 687:692 the side chains 678:692 the side chains of Tyr552 and His553 678:713 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19053759 1 47 theme neurological 243:254 arg1 disorders 256:264 neurological disorders 243:264 neurological disorders 243:264 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 2 48 theme X-ray 383:387 arg1 structures 389:398 X-ray structures 383:398 X-ray structures of four complexes between human GCPII and urea-based inhibitors 383:462 To enhance further development of such compounds, we determined X-ray structures of four complexes between human GCPII and urea-based inhibitors at high resolution. 19053759 5 49 theme P1 815:816 arg1 inhibitors 843:852 the inhibitors 839:852 the inhibitors 839:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 49 theme P1 815:816 arg1 Asn519 897:902 Asn519 897:902 Asn519 897:902 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 49 theme P1 815:816 arg1 Arg534 877:882 Arg534 877:882 Arg534 877:882 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 49 theme P1 815:816 arg1 Arg536 885:890 Arg536 885:890 Arg536 885:890 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 5 49 theme P1 815:816 arg1 group 830:834 the P1 carboxylate group 811:834 the P1 carboxylate group of the inhibitors 811:852 Interactions within the S1 pocket are defined primarily by a network of hydrogen bonds between the P1 carboxylate group of the inhibitors and the side chains of Arg534, Arg536, and Asn519. 19053759 1 50 theme disorders 256:264 arg1 models 233:238 various models 225:238 various models of neurological disorders 225:264 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 0 51 theme human 21:25 arg1 carboxypeptidase 37:52 human glutamate carboxypeptidase II 21:55 human glutamate carboxypeptidase II 21:55 Interactions between human glutamate carboxypeptidase II and urea-based inhibitors: structural characterization. 19053759 1 52 theme carboxypeptidase 167:182 arg1 ligands 146:152 Urea-based, low molecular weight ligands 113:152 ligands 146:152 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 1 52 theme carboxypeptidase 167:182 arg1 agents 291:296 imaging agents 283:296 imaging agents for prostate cancer 283:316 Urea-based, low molecular weight ligands of glutamate carboxypeptidase II (GCPII) have demonstrated efficacy in various models of neurological disorders and can serve as imaging agents for prostate cancer. 19053759 3 53 theme glutarate 521:529 arg1 moiety 531:536 an invariant glutarate moiety 508:536 an invariant glutarate moiety within the S1' pocket of the enzyme 508:572 All ligands demonstrate an invariant glutarate moiety within the S1' pocket of the enzyme. 19053759 4 54 theme inhibitor 613:621 arg1 sites 623:627 P1 and P1' inhibitor sites 602:627 P1 and P1' inhibitor sites 602:627 The ureido linkage between P1 and P1' inhibitor sites interacts with the active-site Zn(1)(2+) ion and the side chains of Tyr552 and His553. 19301871 6 0 from presence 1047:1054 arg1 cavity 1108:1113 the GCPII(E424A) binding cavity 1083:1113 the GCPII(E424A) binding cavity 1083:1113 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 8 1 dep DeltaG 1565:1570 arg1 mol 1606:1608 22(+/-5) kcal x mol 1590:1608 DeltaG(++) approximately 22(+/-5) kcal x mol(-1) 1565:1612 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 8 1 dep DeltaG 1565:1570 arg1 -1 1610:1611 -1 1610:1611 -1 1610:1611 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 9 2 theme reaction 1812:1819 arg1 mechanism 1821:1829 the reaction mechanism 1808:1829 the reaction mechanism of this highly interesting enzyme at the atomic level 1808:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 0 3 theme X-ray 77:81 arg1 crystallography 83:97 X-ray crystallography 77:97 X-ray crystallography 77:97 Reaction mechanism of glutamate carboxypeptidase II revealed by mutagenesis, X-ray crystallography, and computational methods. 19301871 4 4 from involvement 881:891 arg1 hydrolysis 914:923 peptide hydrolysis 906:923 peptide hydrolysis 906:923 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 4 5 theme complete 822:829 arg1 loss 831:834 a complete loss 820:834 a complete loss of catalytic activity 820:856 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 6 6 theme substrate 1070:1078 arg1 presence 1047:1054 The presence 1043:1054 The presence of the intact substrate in the GCPII(E424A) binding cavity 1043:1113 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 6 7 attach presence 1047:1054 arg2 substrate 1070:1078 the intact substrate 1059:1078 the intact substrate 1059:1078 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 6 7 attach presence 1047:1054 arg1 cavity 1108:1113 the GCPII(E424A) binding cavity 1083:1113 the GCPII(E424A) binding cavity 1083:1113 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 4 8 theme peptide 906:912 arg1 hydrolysis 914:923 peptide hydrolysis 906:923 peptide hydrolysis 906:923 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 9 9 from picture 1797:1803 arg1 level 1879:1883 the atomic level 1868:1883 the atomic level 1868:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 4 10 theme catalytic 839:847 arg1 activity 849:856 catalytic activity 839:856 catalytic activity 839:856 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 1 11 dep carboxypeptidase 137:152 arg1 GCPII 158:162 GCPII 158:162 GCPII 158:162 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 1 11 dep carboxypeptidase 137:152 arg1 EC 165:166 EC 3.4.17.21 165:176 EC 3.4.17.21 165:176 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 7 12 theme transition 1364:1373 arg1 states 1375:1380 the transition states 1360:1380 the transition states 1360:1380 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 1 13 theme prostate 274:281 arg1 cancer 283:288 prostate cancer 274:288 prostate cancer 274:288 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 3 14 theme proton 693:698 arg1 Glu424 674:679 Glu424 674:679 Glu424 674:679 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 3 14 theme proton 693:698 arg1 residue 708:714 a putative proton shuttle residue 682:714 a putative proton shuttle residue 682:714 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 9 15 theme detailed 1773:1780 arg1 picture 1797:1803 a detailed and consistent picture 1771:1803 a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level 1771:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 7 16 theme reaction 1408:1415 arg1 barriers 1417:1424 the associated reaction barriers 1393:1424 the associated reaction barriers 1393:1424 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 5 17 from structure 966:974 arg1 complex 995:1001 complex 995:1001 complex with N-Ac-Asp-Glu 995:1019 Additionally, we determined the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu at 1.70 A resolution. 19301871 9 18 theme interesting 1846:1856 arg1 enzyme 1858:1863 this highly interesting enzyme 1834:1863 this highly interesting enzyme 1834:1863 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 3 19 theme shuttle 700:706 arg1 Glu424 674:679 Glu424 674:679 Glu424 674:679 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 3 19 theme shuttle 700:706 arg1 residue 708:714 a putative proton shuttle residue 682:714 a putative proton shuttle residue 682:714 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 9 20 from level 1879:1883 arg1 picture 1797:1803 a detailed and consistent picture 1771:1803 a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level 1771:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 9 20 from level 1879:1883 arg1 mechanism 1821:1829 the reaction mechanism 1808:1829 the reaction mechanism of this highly interesting enzyme at the atomic level 1808:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 4 21 theme Kinetic 746:752 arg1 analysis 754:761 Kinetic analysis 746:761 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate 746:809 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 7 22 theme mechanics 1296:1304 arg1 barriers 1417:1424 the associated reaction barriers 1393:1424 the associated reaction barriers 1393:1424 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 22 theme mechanics 1296:1304 arg1 QM/MM 1320:1324 QM/MM 1320:1324 QM/MM 1320:1324 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 22 theme mechanics 1296:1304 arg1 calculations 1306:1317 the combined quantum mechanics/molecular mechanics calculations 1255:1317 the combined quantum mechanics/molecular mechanics calculations (QM/MM) 1255:1325 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 0 23 theme computational 104:116 arg1 methods 118:124 computational methods 104:124 computational methods 104:124 Reaction mechanism of glutamate carboxypeptidase II revealed by mutagenesis, X-ray crystallography, and computational methods. 19301871 6 24 theme interactions 1199:1210 arg1 analysis 1168:1175 a detailed analysis 1157:1175 a detailed analysis of GCPII/N-Ac-Asp-Glu interactions 1157:1210 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 0 25 theme Reaction 0:7 arg1 mechanism 9:17 Reaction mechanism 0:17 Reaction mechanism of glutamate carboxypeptidase II 0:50 Reaction mechanism of glutamate carboxypeptidase II revealed by mutagenesis, X-ray crystallography, and computational methods. 19301871 8 26 theme good 1629:1632 arg1 agreement 1634:1642 a good agreement 1627:1642 a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)) 1627:1729 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 5 27 theme A 1029:1029 arg1 resolution 1031:1040 1.70 A resolution 1024:1040 1.70 A resolution 1024:1040 Additionally, we determined the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu at 1.70 A resolution. 19301871 3 28 theme putative 684:691 arg1 Glu424 674:679 Glu424 674:679 Glu424 674:679 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 3 28 theme putative 684:691 arg1 residue 708:714 a putative proton shuttle residue 682:714 a putative proton shuttle residue 682:714 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 6 29 theme GCPII/N-Ac-Asp-Glu 1180:1197 arg1 interactions 1199:1210 GCPII/N-Ac-Asp-Glu interactions 1180:1210 GCPII/N-Ac-Asp-Glu interactions 1180:1210 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 8 30 theme reaction 1527:1534 arg1 barrier 1536:1542 the reaction barrier 1523:1542 the reaction barrier 1523:1542 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 8 30 theme reaction 1527:1534 arg1 DeltaG 1565:1570 DeltaG 1565:1570 DeltaG(++) approximately 22(+/-5) kcal x mol(-1) 1565:1612 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 6 31 theme E424A 1093:1097 arg1 cavity 1108:1113 the GCPII(E424A) binding cavity 1083:1113 the GCPII(E424A) binding cavity 1083:1113 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 5 32 with complex 995:1001 arg1 N-Ac-Asp-Glu 1008:1019 N-Ac-Asp-Glu 1008:1019 N-Ac-Asp-Glu 1008:1019 Additionally, we determined the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu at 1.70 A resolution. 19301871 7 33 theme quantum 1268:1274 arg1 barriers 1417:1424 the associated reaction barriers 1393:1424 the associated reaction barriers 1393:1424 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 33 theme quantum 1268:1274 arg1 QM/MM 1320:1324 QM/MM 1320:1324 QM/MM 1320:1324 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 33 theme quantum 1268:1274 arg1 calculations 1306:1317 the combined quantum mechanics/molecular mechanics calculations 1255:1317 the combined quantum mechanics/molecular mechanics calculations (QM/MM) 1255:1325 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 2 34 theme nervous 470:476 arg1 system 478:483 the mammalian nervous system 456:483 the mammalian nervous system 456:483 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 7 35 theme associated 1397:1406 arg1 barriers 1417:1424 the associated reaction barriers 1393:1424 the associated reaction barriers 1393:1424 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 8 36 with agreement 1634:1642 arg1 constant 1691:1698 the experimentally observed reaction rate constant 1649:1698 the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)) 1649:1729 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 9 37 theme atomic 1872:1877 arg1 level 1879:1883 the atomic level 1868:1883 the atomic level 1868:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 7 38 theme mechanics/molecular 1276:1294 arg1 barriers 1417:1424 the associated reaction barriers 1393:1424 the associated reaction barriers 1393:1424 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 38 theme mechanics/molecular 1276:1294 arg1 QM/MM 1320:1324 QM/MM 1320:1324 QM/MM 1320:1324 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 38 theme mechanics/molecular 1276:1294 arg1 calculations 1306:1317 the combined quantum mechanics/molecular mechanics calculations 1255:1317 the combined quantum mechanics/molecular mechanics calculations (QM/MM) 1255:1325 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 0 39 theme glutamate 22:30 arg1 carboxypeptidase 32:47 glutamate carboxypeptidase II 22:50 glutamate carboxypeptidase II 22:50 Reaction mechanism of glutamate carboxypeptidase II revealed by mutagenesis, X-ray crystallography, and computational methods. 19301871 3 40 theme peptide 571:577 arg1 hydrolysis 579:588 peptide hydrolysis 571:588 peptide hydrolysis by GCPII in detail 571:607 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 1 41 theme zinc-dependent 184:197 arg1 carboxypeptidase 137:152 Glutamate carboxypeptidase II 127:155 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) 127:177 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 1 41 theme zinc-dependent 184:197 arg1 exopeptidase 199:210 a zinc-dependent exopeptidase 182:210 a zinc-dependent exopeptidase 182:210 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 4 42 theme direct 874:879 arg1 involvement 881:891 the direct involvement 870:891 the direct involvement of Glu424 in peptide hydrolysis 870:923 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 3 43 theme [GCPII 649:654 arg1 mutant 627:632 a mutant 625:632 a mutant 625:632 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 2 44 theme N-acetyl-l-aspartyl-l-glutamate 309:339 arg1 hydrolysis 295:304 The hydrolysis 291:304 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, 291:402 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 3 45 theme human 637:641 arg1 [GCPII 649:654 human GCPII [GCPII 637:654 human GCPII [GCPII(E424A)] 637:662 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 3 45 theme human 637:641 arg1 E424A 656:660 E424A 656:660 E424A 656:660 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 7 46 theme combined 1259:1266 arg1 barriers 1417:1424 the associated reaction barriers 1393:1424 the associated reaction barriers 1393:1424 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 46 theme combined 1259:1266 arg1 QM/MM 1320:1324 QM/MM 1320:1324 QM/MM 1320:1324 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 7 46 theme combined 1259:1266 arg1 calculations 1306:1317 the combined quantum mechanics/molecular mechanics calculations 1255:1317 the combined quantum mechanics/molecular mechanics calculations (QM/MM) 1255:1325 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 2 47 theme exact 494:498 arg1 mechanism 500:508 the exact mechanism 490:508 the exact mechanism of this reaction 490:525 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 2 48 theme natural 361:367 arg1 N-acetyl-l-aspartyl-l-glutamate 309:339 N-acetyl-l-aspartyl-l-glutamate 309:339 N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu) 309:354 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 2 48 theme natural 361:367 arg1 substrate 380:388 the natural dipeptidic substrate 357:388 the natural dipeptidic substrate of the GCPII 357:401 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 4 49 theme Glu424 896:901 arg1 involvement 881:891 the direct involvement 870:891 the direct involvement of Glu424 in peptide hydrolysis 870:923 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 5 50 theme GCPII 979:983 arg1 structure 966:974 the crystal structure 954:974 the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu 954:1019 Additionally, we determined the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu at 1.70 A resolution. 19301871 9 51 theme mechanism 1821:1829 arg1 picture 1797:1803 a detailed and consistent picture 1771:1803 a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level 1771:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 2 52 theme cellular 430:437 arg1 signaling 439:447 cellular signaling 430:447 cellular signaling within the mammalian nervous system 430:483 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 0 53 theme carboxypeptidase 32:47 arg1 mechanism 9:17 Reaction mechanism 0:17 Reaction mechanism of glutamate carboxypeptidase II 0:50 Reaction mechanism of glutamate carboxypeptidase II revealed by mutagenesis, X-ray crystallography, and computational methods. 19301871 7 54 theme detailed 1440:1447 arg1 information 1449:1459 detailed information 1440:1459 detailed information concerning the GCPII reaction mechanism 1440:1499 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 1 55 dep important 219:227 arg1 therapeutic 229:239 therapeutic 229:239 therapeutic 229:239 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 2 56 theme mammalian 460:468 arg1 system 478:483 the mammalian nervous system 456:483 the mammalian nervous system 456:483 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 8 57 theme reaction 1677:1684 arg1 constant 1691:1698 the experimentally observed reaction rate constant 1649:1698 the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)) 1649:1729 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 3 58 theme GCPII 643:647 arg1 [GCPII 649:654 human GCPII [GCPII 637:654 human GCPII [GCPII(E424A)] 637:662 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 3 58 theme GCPII 643:647 arg1 E424A 656:660 E424A 656:660 E424A 656:660 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 8 59 theme observed 1668:1675 arg1 constant 1691:1698 the experimentally observed reaction rate constant 1649:1698 the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)) 1649:1729 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 2 60 theme dipeptidic 369:378 arg1 N-acetyl-l-aspartyl-l-glutamate 309:339 N-acetyl-l-aspartyl-l-glutamate 309:339 N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu) 309:354 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 2 60 theme dipeptidic 369:378 arg1 substrate 380:388 the natural dipeptidic substrate 357:388 the natural dipeptidic substrate of the GCPII 357:401 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 8 61 theme kcal 1599:1602 arg1 mol 1606:1608 22(+/-5) kcal x mol 1590:1608 DeltaG(++) approximately 22(+/-5) kcal x mol(-1) 1565:1612 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 8 61 theme kcal 1599:1602 arg1 -1 1610:1611 -1 1610:1611 -1 1610:1611 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 4 62 theme activity 849:856 arg1 loss 831:834 a complete loss 820:834 a complete loss of catalytic activity 820:856 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 6 63 theme GCPII 1087:1091 arg1 cavity 1108:1113 the GCPII(E424A) binding cavity 1083:1113 the GCPII(E424A) binding cavity 1083:1113 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 1 64 theme Glutamate 127:135 arg1 carboxypeptidase 137:152 Glutamate carboxypeptidase II 127:155 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) 127:177 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 1 64 theme Glutamate 127:135 arg1 exopeptidase 199:210 a zinc-dependent exopeptidase 182:210 a zinc-dependent exopeptidase 182:210 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 8 65 theme x 1604:1604 arg1 mol 1606:1608 22(+/-5) kcal x mol 1590:1608 DeltaG(++) approximately 22(+/-5) kcal x mol(-1) 1565:1612 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 8 65 theme x 1604:1604 arg1 -1 1610:1611 -1 1610:1611 -1 1610:1611 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 6 66 theme detailed 1159:1166 arg1 analysis 1168:1175 a detailed analysis 1157:1175 a detailed analysis of GCPII/N-Ac-Asp-Glu interactions 1157:1210 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 6 67 theme intact 1063:1068 arg1 substrate 1070:1078 the intact substrate 1059:1078 the intact substrate 1059:1078 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 3 68 from hydrolysis 579:588 arg1 detail 602:607 detail 602:607 detail 602:607 To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. 19301871 1 69 theme important 219:227 arg1 target 241:246 an important therapeutic target 216:246 an important therapeutic target for neurodegeneration and prostate cancer 216:288 Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. 19301871 8 70 theme rate 1686:1689 arg1 constant 1691:1698 the experimentally observed reaction rate constant 1649:1698 the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)) 1649:1729 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 7 71 theme GCPII 1476:1480 arg1 mechanism 1491:1499 the GCPII reaction mechanism 1472:1499 the GCPII reaction mechanism 1472:1499 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 5 72 theme crystal 958:964 arg1 structure 966:974 the crystal structure 954:974 the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu 954:1019 Additionally, we determined the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu at 1.70 A resolution. 19301871 9 73 theme enzyme 1858:1863 arg1 mechanism 1821:1829 the reaction mechanism 1808:1829 the reaction mechanism of this highly interesting enzyme at the atomic level 1808:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 7 74 theme experimental 1217:1228 arg1 data 1230:1233 The experimental data 1213:1233 The experimental data 1213:1233 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 8 75 dep k 1701:1701 arg1 -1 1726:1727 -1 1726:1727 -1 1726:1727 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 8 75 dep k 1701:1701 arg1 s 1724:1724 1 s 1722:1724 k(cat) approximately 1 s(-1) 1701:1728 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 9 76 theme consistent 1786:1795 arg1 picture 1797:1803 a detailed and consistent picture 1771:1803 a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level 1771:1883 Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. 19301871 2 77 theme reaction 518:525 arg1 mechanism 500:508 the exact mechanism 490:508 the exact mechanism of this reaction 490:525 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 7 78 theme reaction 1482:1489 arg1 mechanism 1491:1499 the GCPII reaction mechanism 1472:1499 the GCPII reaction mechanism 1472:1499 The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. 19301871 6 79 theme kinetic 1133:1139 arg1 data 1141:1144 our kinetic data 1129:1144 our kinetic data 1129:1144 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 2 80 theme GCPII 397:401 arg1 N-acetyl-l-aspartyl-l-glutamate 309:339 N-acetyl-l-aspartyl-l-glutamate 309:339 N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu) 309:354 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 2 80 theme GCPII 397:401 arg1 substrate 380:388 the natural dipeptidic substrate 357:388 the natural dipeptidic substrate of the GCPII 357:401 The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp-Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. 19301871 6 81 theme binding 1100:1106 arg1 cavity 1108:1113 the GCPII(E424A) binding cavity 1083:1113 the GCPII(E424A) binding cavity 1083:1113 The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. 19301871 4 82 theme GCPII 766:770 arg1 analysis 754:761 Kinetic analysis 746:761 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate 746:809 Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. 19301871 8 83 dep constant 1691:1698 arg1 k 1701:1701 k 1701:1701 k(cat) approximately 1 s(-1) 1701:1728 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 19301871 8 83 dep constant 1691:1698 arg1 cat 1703:1705 cat 1703:1705 cat 1703:1705 The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). 31416931 2 0 theme angstroms 485:493 arg1 resolutions 459:469 resolutions 459:469 resolutions of 2.6 to 3.3 angstroms 459:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 4 1 theme acyl 907:910 arg1 chain 912:916 the acyl chain 903:916 the acyl chain 903:916 The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. 31416931 1 2 theme lipid 245:249 arg1 asymmetry 251:259 lipid asymmetry 245:259 lipid asymmetry 245:259 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 2 3 theme translocation 518:530 arg1 cycle 532:536 the lipid translocation cycle 508:536 the lipid translocation cycle of this P4-ATPase 508:554 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 4 theme first 746:750 arg1 helices 777:783 the first and second transmembrane helices 742:783 the first and second transmembrane helices 742:783 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 5 5 theme flippase 1007:1014 arg1 mechanism 1016:1024 the flippase mechanism 1003:1024 the flippase mechanism 1003:1024 These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. 31416931 2 6 theme lipid 512:516 arg1 cycle 532:536 the lipid translocation cycle 508:536 the lipid translocation cycle of this P4-ATPase 508:554 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 2 7 from resolutions 459:469 arg1 heterocomplex 442:454 the human ATP8A1-CDC50a heterocomplex 418:454 the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 418:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 2 7 from resolutions 459:469 arg1 intermediates 401:413 six distinct intermediates 388:413 six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 388:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 5 8 theme mechanism 1016:1024 arg1 understanding 986:998 our understanding 982:998 our understanding of the flippase mechanism 982:1024 These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. 31416931 5 8 theme mechanism 1016:1024 arg1 mutants 1053:1059 the disease-associated mutants 1030:1059 the disease-associated mutants of P4-ATPases 1030:1073 These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. 31416931 4 9 theme hydrophilic 878:888 arg1 cleft 890:894 the hydrophilic cleft 874:894 the hydrophilic cleft 874:894 The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. 31416931 0 10 theme Cryo-EM 0:6 arg1 structures 8:17 Cryo-EM structures 0:17 Cryo-EM structures 0:17 Cryo-EM structures capture the transport cycle of the P4-ATPase flippase. 31416931 3 11 from site 673:676 arg1 domain 701:706 the phosphorylation domain 681:706 the phosphorylation domain 681:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 1 12 theme eukaryotic 77:86 arg1 membranes 88:96 eukaryotic membranes 77:96 eukaryotic membranes 77:96 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 2 13 theme heterocomplex 442:454 arg1 intermediates 401:413 six distinct intermediates 388:413 six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 388:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 14 theme helices 777:783 arg1 shifts 732:737 lateral shifts 724:737 lateral shifts of the first and second transmembrane helices 724:783 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 2 15 theme microscopy 363:372 arg1 structures 374:383 the cryo-electron microscopy structures 345:383 the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 345:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 2 16 theme ATP8A1-CDC50a 428:440 arg1 heterocomplex 442:454 the human ATP8A1-CDC50a heterocomplex 418:454 the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 418:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 17 theme ATP-dependent 557:569 arg1 phosphorylation 571:585 ATP-dependent phosphorylation 557:585 ATP-dependent phosphorylation 557:585 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 5 18 theme P4-ATPases 1064:1073 arg1 understanding 986:998 our understanding 982:998 our understanding of the flippase mechanism 982:1024 These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. 31416931 5 18 theme P4-ATPases 1064:1073 arg1 mutants 1053:1059 the disease-associated mutants 1030:1059 the disease-associated mutants of P4-ATPases 1030:1073 These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. 31416931 2 19 theme human 422:426 arg1 heterocomplex 442:454 the human ATP8A1-CDC50a heterocomplex 418:454 the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 418:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 20 theme second 756:761 arg1 helices 777:783 the first and second transmembrane helices 742:783 the first and second transmembrane helices 742:783 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 2 21 theme P4-ATPase 546:554 arg1 cycle 532:536 the lipid translocation cycle 508:536 the lipid translocation cycle of this P4-ATPase 508:554 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 22 theme transmembrane 763:775 arg1 helices 777:783 the first and second transmembrane helices 742:783 the first and second transmembrane helices 742:783 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 4 23 theme phospholipid 835:846 arg1 group 853:857 The phospholipid head group 831:857 The phospholipid head group 831:857 The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. 31416931 1 24 theme phospholipids 182:194 arg1 translocation 165:177 the translocation 161:177 the translocation of phospholipids from the outer to the inner leaflet 161:230 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 2 25 theme intermediates 401:413 arg1 structures 374:383 the cryo-electron microscopy structures 345:383 the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 345:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 0 26 theme transport 31:39 arg1 cycle 41:45 the transport cycle 27:45 the transport cycle of the P4-ATPase flippase 27:71 Cryo-EM structures capture the transport cycle of the P4-ATPase flippase. 31416931 1 27 theme membrane 284:291 arg1 trafficking 293:303 membrane trafficking 284:303 membrane trafficking 284:303 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 1 28 from outer 205:209 arg1 translocation 165:177 the translocation 161:177 the translocation of phospholipids from the outer to the inner leaflet 161:230 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 2 29 theme distinct 392:399 arg1 intermediates 401:413 six distinct intermediates 388:413 six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 388:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 30 theme phosphatidylserine 803:820 arg1 binding 822:828 phosphatidylserine binding 803:828 phosphatidylserine binding 803:828 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 2 31 dep angstroms 485:493 arg1 to 478:479 to 478:479 to 478:479 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 3 32 theme phosphorylation 657:671 arg1 site 673:676 the phosphorylation site 653:676 the phosphorylation site in the phosphorylation domain 653:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 3 33 theme large 597:601 arg1 movement 614:621 a large rotational movement 595:621 a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain 595:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 3 34 theme rotational 603:612 arg1 movement 614:621 a large rotational movement 595:621 a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain 595:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 1 35 theme P-type 107:112 arg1 triphosphatases 124:138 P-type adenosine triphosphatases 107:138 type IV P-type adenosine triphosphatases (P4-ATPases) 99:151 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 1 36 theme signaling 309:317 arg1 pathways 319:326 signaling pathways 309:326 signaling pathways 309:326 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 4 37 theme lipid 940:944 arg1 environment 946:956 the lipid environment 936:956 the lipid environment 936:956 The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. 31416931 3 38 theme domain 639:644 arg1 movement 614:621 a large rotational movement 595:621 a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain 595:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 1 39 theme adenosine 114:122 arg1 triphosphatases 124:138 P-type adenosine triphosphatases 107:138 type IV P-type adenosine triphosphatases (P4-ATPases) 99:151 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 5 40 theme disease-associated 1034:1051 arg1 mutants 1053:1059 the disease-associated mutants 1030:1059 the disease-associated mutants of P4-ATPases 1030:1073 These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. 31416931 2 41 from intermediates 401:413 arg1 resolutions 459:469 resolutions 459:469 resolutions of 2.6 to 3.3 angstroms 459:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 1 42 theme inner 218:222 arg1 leaflet 224:230 the inner leaflet 214:230 the inner leaflet 214:230 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 31416931 0 43 theme flippase 64:71 arg1 cycle 41:45 the transport cycle 27:45 the transport cycle of the P4-ATPase flippase 27:71 Cryo-EM structures capture the transport cycle of the P4-ATPase flippase. 31416931 3 44 theme phosphorylation 685:699 arg1 domain 701:706 the phosphorylation domain 681:706 the phosphorylation domain 681:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 4 45 theme head 848:851 arg1 group 853:857 The phospholipid head group 831:857 The phospholipid head group 831:857 The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. 31416931 2 46 theme cryo-electron 349:361 arg1 microscopy 363:372 cryo-electron microscopy 349:372 the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms 345:493 Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. 31416931 0 47 theme P4-ATPase 54:62 arg1 flippase 64:71 the P4-ATPase flippase 50:71 the P4-ATPase flippase 50:71 Cryo-EM structures capture the transport cycle of the P4-ATPase flippase. 31416931 3 48 theme lateral 724:730 arg1 shifts 732:737 lateral shifts 724:737 lateral shifts of the first and second transmembrane helices 724:783 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 3 49 theme actuator 630:637 arg1 domain 639:644 the actuator domain 626:644 the actuator domain around the phosphorylation site in the phosphorylation domain 626:706 ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. 31416931 1 50 dep type 99:102 arg1 triphosphatases 124:138 P-type adenosine triphosphatases 107:138 type IV P-type adenosine triphosphatases (P4-ATPases) 99:151 In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. 14981520 2 0 theme paraparesis 434:444 arg1 form 407:410 a rare form 400:410 a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs 400:595 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 2 0 theme paraparesis 434:444 arg1 syndrome 373:380 Silver syndrome 366:380 Silver syndrome (OMIM #270685) 366:395 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 0 1 from mutations 22:30 arg1 BSCL2 35:39 BSCL2 35:39 BSCL2 35:39 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 3 2 theme incomplete 677:686 arg1 penetrance 688:697 incomplete penetrance 677:697 incomplete penetrance 677:697 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 5 3 with families 881:888 arg1 characteristic 907:920 a phenotype characteristic 895:920 a phenotype characteristic of dHMN or Silver syndrome 895:947 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 9 4 dep substitutions 1459:1471 arg1 substitutions 1459:1471 The amino acid substitutions 1444:1471 The amino acid substitutions N88S and S90L 1444:1485 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 9 4 dep substitutions 1459:1471 arg1 S90L 1482:1485 S90L 1482:1485 S90L 1482:1485 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 9 4 dep substitutions 1459:1471 arg1 N88S 1473:1476 N88S 1473:1476 N88S 1473:1476 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 0 5 theme motor 79:83 arg1 neuropathy 85:94 distal hereditary motor neuropathy 61:94 distal hereditary motor neuropathy 61:94 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 3 6 theme most 618:621 arg1 forms 623:627 most forms 618:627 most forms of dHMN 618:635 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 3 7 from penetrance 688:697 arg1 expression 735:744 clinical expression 726:744 clinical expression 726:744 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 6 8 theme Berardinelli-Seip 1016:1032 arg1 BSCL2 1060:1064 BSCL2 1060:1064 BSCL2 1060:1064 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 8 theme Berardinelli-Seip 1016:1032 arg1 lipodystrophy 1045:1057 the gene Berardinelli-Seip congenital lipodystrophy 1007:1057 the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) 1007:1065 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 9 9 theme aggregate 1532:1540 arg1 formation 1542:1550 aggregate formation 1532:1550 aggregate formation leading to neurodegeneration 1532:1579 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 0 10 theme Silver 100:105 arg1 syndrome 107:114 Silver syndrome 100:114 Silver syndrome 100:114 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 3 11 from variability 711:721 arg1 expression 735:744 clinical expression 726:744 clinical expression 726:744 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 2 12 theme lower 585:589 arg1 limbs 591:595 the lower limbs 581:595 the lower limbs 581:595 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 6 13 theme heterozygous 1086:1097 arg1 mutations 1108:1116 two heterozygous missense mutations 1082:1116 two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L 1082:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 1 14 theme disorders 236:244 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 14 theme disorders 236:244 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 14 theme disorders 236:244 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 14 theme disorders 236:244 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 7 15 theme Null 1175:1178 arg1 mutations 1180:1188 Null mutations 1175:1188 Null mutations 1175:1188 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 15 theme Null 1175:1178 arg1 seipin 1226:1231 the protein seipin 1214:1231 the protein seipin 1214:1231 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 8 16 theme reticulum 1428:1436 arg1 protein 1401:1407 an integral membrane protein 1380:1407 an integral membrane protein of the endoplasmic reticulum (ER) 1380:1441 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 8 16 theme reticulum 1428:1436 arg1 seipin 1370:1375 seipin 1370:1375 seipin 1370:1375 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 5 17 theme additional 870:879 arg1 families 881:888 16 additional families 867:888 16 additional families with a phenotype characteristic of dHMN or Silver syndrome 867:947 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 1 18 theme distal 340:345 arg1 part 347:350 the distal part 336:350 the distal part of the limbs 336:363 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 7 19 theme recessive 1288:1296 arg1 lipodystrophy 1327:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy 1278:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700) 1278:1354 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 19 theme recessive 1288:1296 arg1 269700 1348:1353 OMIM #269700 1342:1353 OMIM #269700 1342:1353 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 20 theme OMIM 1342:1345 arg1 lipodystrophy 1327:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy 1278:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700) 1278:1354 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 20 theme OMIM 1342:1345 arg1 269700 1348:1353 OMIM #269700 1342:1353 OMIM #269700 1342:1353 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 21 theme protein 1218:1224 arg1 seipin 1226:1231 the protein seipin 1214:1231 the protein seipin 1214:1231 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 21 theme protein 1218:1224 arg1 mutations 1180:1188 Null mutations 1175:1188 Null mutations 1175:1188 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 1 22 theme distal 162:167 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 22 theme distal 162:167 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 22 theme distal 162:167 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 22 theme distal 162:167 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 22 theme distal 162:167 arg1 182960 200:205 OMIM #182960 194:205 OMIM #182960 194:205 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 4 23 theme Austrian 772:779 arg1 family 781:786 an Austrian family 769:786 an Austrian family 769:786 A genome-wide scan in an Austrian family with dHMN-V (ref. 14981520 7 24 theme # 1347:1347 arg1 lipodystrophy 1327:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy 1278:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700) 1278:1354 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 24 theme # 1347:1347 arg1 269700 1348:1353 OMIM #269700 1342:1353 OMIM #269700 1342:1353 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 1 25 theme spinal 169:174 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 25 theme spinal 169:174 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 25 theme spinal 169:174 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 25 theme spinal 169:174 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 25 theme spinal 169:174 arg1 182960 200:205 OMIM #182960 194:205 OMIM #182960 194:205 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 0 26 theme missense 13:20 arg1 mutations 22:30 Heterozygous missense mutations 0:30 Heterozygous missense mutations in BSCL2 0:39 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 2 27 theme spastic 426:432 arg1 paraparesis 434:444 hereditary spastic paraparesis 415:444 hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) 415:483 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 2 28 theme hands 553:557 arg1 amyotrophy 535:544 amyotrophy 535:544 amyotrophy of the hands 535:557 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 7 29 from mutations 1180:1188 arg1 BSCL2 1193:1197 BSCL2 1193:1197 BSCL2 1193:1197 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 9 30 theme acid 1454:1457 arg1 substitutions 1459:1471 The amino acid substitutions 1444:1471 The amino acid substitutions N88S and S90L 1444:1485 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 9 30 theme acid 1454:1457 arg1 S90L 1482:1485 S90L 1482:1485 S90L 1482:1485 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 9 30 theme acid 1454:1457 arg1 N88S 1473:1476 N88S 1473:1476 N88S 1473:1476 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 8 31 theme endoplasmic 1416:1426 arg1 reticulum 1428:1436 the endoplasmic reticulum 1412:1436 the endoplasmic reticulum (ER) 1412:1441 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 8 31 theme endoplasmic 1416:1426 arg1 ER 1439:1440 ER 1439:1440 ER 1439:1440 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 1 32 theme muscular 176:183 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 32 theme muscular 176:183 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 32 theme muscular 176:183 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 32 theme muscular 176:183 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 32 theme muscular 176:183 arg1 182960 200:205 OMIM #182960 194:205 OMIM #182960 194:205 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 0 33 theme Heterozygous 0:11 arg1 mutations 22:30 Heterozygous missense mutations 0:30 Heterozygous missense mutations in BSCL2 0:39 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 9 34 gly glycosylation 1494:1506 arg1 seipin 1511:1516 seipin 1511:1516 seipin 1511:1516 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 6 35 theme critical 969:976 arg1 region 978:983 the critical region 965:983 the critical region to 1 Mb 965:991 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 8 36 theme integral 1383:1390 arg1 protein 1401:1407 an integral membrane protein 1380:1407 an integral membrane protein of the endoplasmic reticulum (ER) 1380:1441 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 8 36 theme integral 1383:1390 arg1 seipin 1370:1375 seipin 1370:1375 seipin 1370:1375 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 7 37 theme Berardinelli-Seip 1298:1314 arg1 lipodystrophy 1327:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy 1278:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700) 1278:1354 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 37 theme Berardinelli-Seip 1298:1314 arg1 269700 1348:1353 OMIM #269700 1342:1353 OMIM #269700 1342:1353 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 38 theme autosomal 1278:1286 arg1 lipodystrophy 1327:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy 1278:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700) 1278:1354 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 38 theme autosomal 1278:1286 arg1 269700 1348:1353 OMIM #269700 1342:1353 OMIM #269700 1342:1353 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 5 39 theme locus 831:835 arg1 SPG17 837:841 the locus SPG17 827:841 the locus SPG17 827:841 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 7 40 theme congenital 1316:1325 arg1 lipodystrophy 1327:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy 1278:1339 autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700) 1278:1354 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 7 40 theme congenital 1316:1325 arg1 269700 1348:1353 OMIM #269700 1342:1353 OMIM #269700 1342:1353 Null mutations in BSCL2, which encodes the protein seipin, were previously shown to be associated with autosomal recessive Berardinelli-Seip congenital lipodystrophy (OMIM #269700). 14981520 1 41 theme OMIM 194:197 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 41 theme OMIM 194:197 arg1 182960 200:205 OMIM #182960 194:205 OMIM #182960 194:205 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 2 42 theme hereditary 415:424 arg1 paraparesis 434:444 hereditary spastic paraparesis 415:444 hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) 415:483 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 1 43 theme exclusive 273:281 arg1 degeneration 283:294 an almost exclusive degeneration 263:294 an almost exclusive degeneration of motor nerve fibers 263:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 44 from group 227:231 arg1 part 347:350 the distal part 336:350 the distal part of the limbs 336:363 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 45 theme limbs 359:363 arg1 part 347:350 the distal part 336:350 the distal part of the limbs 336:363 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 5 46 theme dHMN 925:928 arg1 characteristic 907:920 a phenotype characteristic 895:920 a phenotype characteristic of dHMN or Silver syndrome 895:947 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 1 47 theme # 199:199 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 47 theme # 199:199 arg1 182960 200:205 OMIM #182960 194:205 OMIM #182960 194:205 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 2 48 theme Silver 366:371 arg1 form 407:410 a rare form 400:410 a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs 400:595 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 2 48 theme Silver 366:371 arg1 syndrome 373:380 Silver syndrome 366:380 Silver syndrome (OMIM #270685) 366:395 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 6 49 dep substitutions 1146:1158 arg1 S90L 1169:1172 S90L 1169:1172 S90L 1169:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 49 dep substitutions 1146:1158 arg1 N88S 1160:1163 N88S 1160:1163 N88S 1160:1163 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 49 dep substitutions 1146:1158 arg1 substitutions 1146:1158 the amino acid substitutions 1131:1158 the amino acid substitutions N88S and S90L 1131:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 2 50 dep syndrome 373:380 arg1 # 388:388 # 388:388 # 388:388 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 3 51 theme broad 705:709 arg1 variability 711:721 a broad variability 703:721 a broad variability in clinical expression 703:744 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 5 52 theme Silver 933:938 arg1 syndrome 940:947 Silver syndrome 933:947 Silver syndrome 933:947 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 1 53 theme Distal 117:122 arg1 dHMN 153:156 dHMN 153:156 dHMN 153:156 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 53 theme Distal 117:122 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 53 theme Distal 117:122 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 53 theme Distal 117:122 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 53 theme Distal 117:122 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 2 54 theme rare 402:405 arg1 form 407:410 a rare form 400:410 a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs 400:595 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 2 54 theme rare 402:405 arg1 syndrome 373:380 Silver syndrome 366:380 Silver syndrome (OMIM #270685) 366:395 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 1 55 theme motor 299:303 arg1 fibers 311:316 motor nerve fibers 299:316 motor nerve fibers 299:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 2 56 theme legs 512:515 arg1 spasticity 494:503 spasticity 494:503 spasticity of the legs 494:515 Silver syndrome (OMIM #270685) is a rare form of hereditary spastic paraparesis mapped to chromosome 11q12-q14 (SPG17) in which spasticity of the legs is accompanied by amyotrophy of the hands and occasionally also the lower limbs. 14981520 5 57 theme syndrome 940:947 arg1 characteristic 907:920 a phenotype characteristic 895:920 a phenotype characteristic of dHMN or Silver syndrome 895:947 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 1 58 theme hereditary 124:133 arg1 dHMN 153:156 dHMN 153:156 dHMN 153:156 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 58 theme hereditary 124:133 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 58 theme hereditary 124:133 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 58 theme hereditary 124:133 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 58 theme hereditary 124:133 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 59 theme nerve 305:309 arg1 fibers 311:316 motor nerve fibers 299:316 motor nerve fibers 299:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 9 60 theme seipin 1511:1516 arg1 glycosylation 1494:1506 glycosylation 1494:1506 glycosylation of seipin 1494:1516 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 6 61 theme acid 1141:1144 arg1 S90L 1169:1172 S90L 1169:1172 S90L 1169:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 61 theme acid 1141:1144 arg1 N88S 1160:1163 N88S 1160:1163 N88S 1160:1163 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 61 theme acid 1141:1144 arg1 substitutions 1146:1158 the amino acid substitutions 1131:1158 the amino acid substitutions N88S and S90L 1131:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 1 62 theme motor 135:139 arg1 dHMN 153:156 dHMN 153:156 dHMN 153:156 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 62 theme motor 135:139 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 62 theme motor 135:139 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 62 theme motor 135:139 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 62 theme motor 135:139 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 9 63 theme amino 1448:1452 arg1 substitutions 1459:1471 The amino acid substitutions 1444:1471 The amino acid substitutions N88S and S90L 1444:1485 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 9 63 theme amino 1448:1452 arg1 S90L 1482:1485 S90L 1482:1485 S90L 1482:1485 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 9 63 theme amino 1448:1452 arg1 N88S 1473:1476 N88S 1473:1476 N88S 1473:1476 The amino acid substitutions N88S and S90L affect glycosylation of seipin and result in aggregate formation leading to neurodegeneration. 14981520 6 64 theme amino 1135:1139 arg1 S90L 1169:1172 S90L 1169:1172 S90L 1169:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 64 theme amino 1135:1139 arg1 N88S 1160:1163 N88S 1160:1163 N88S 1160:1163 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 64 theme amino 1135:1139 arg1 substitutions 1146:1158 the amino acid substitutions 1131:1158 the amino acid substitutions N88S and S90L 1131:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 5 65 theme phenotype 897:905 arg1 characteristic 907:920 a phenotype characteristic 895:920 a phenotype characteristic of dHMN or Silver syndrome 895:947 4) showed linkage to the locus SPG17, which was confirmed in 16 additional families with a phenotype characteristic of dHMN or Silver syndrome. 14981520 8 66 theme membrane 1392:1399 arg1 protein 1401:1407 an integral membrane protein 1380:1407 an integral membrane protein of the endoplasmic reticulum (ER) 1380:1441 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 8 66 theme membrane 1392:1399 arg1 seipin 1370:1375 seipin 1370:1375 seipin 1370:1375 We show that seipin is an integral membrane protein of the endoplasmic reticulum (ER). 14981520 6 67 theme congenital 1034:1043 arg1 BSCL2 1060:1064 BSCL2 1060:1064 BSCL2 1060:1064 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 67 theme congenital 1034:1043 arg1 lipodystrophy 1045:1057 the gene Berardinelli-Seip congenital lipodystrophy 1007:1057 the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) 1007:1065 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 68 theme gene 1011:1014 arg1 BSCL2 1060:1064 BSCL2 1060:1064 BSCL2 1060:1064 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 6 68 theme gene 1011:1014 arg1 lipodystrophy 1045:1057 the gene Berardinelli-Seip congenital lipodystrophy 1007:1057 the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) 1007:1065 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 0 69 theme hereditary 68:77 arg1 neuropathy 85:94 distal hereditary motor neuropathy 61:94 distal hereditary motor neuropathy 61:94 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 3 70 theme Silver 598:603 arg1 syndrome 605:612 Silver syndrome 598:612 Silver syndrome 598:612 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 3 71 theme clinical 726:733 arg1 expression 735:744 clinical expression 726:744 clinical expression 726:744 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 0 72 theme distal 61:66 arg1 neuropathy 85:94 distal hereditary motor neuropathy 61:94 distal hereditary motor neuropathy 61:94 Heterozygous missense mutations in BSCL2 are associated with distal hereditary motor neuropathy and Silver syndrome. 14981520 1 73 theme fibers 311:316 arg1 degeneration 283:294 an almost exclusive degeneration 263:294 an almost exclusive degeneration of motor nerve fibers 263:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 6 74 theme missense 1099:1106 arg1 mutations 1108:1116 two heterozygous missense mutations 1082:1116 two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L 1082:1172 After refining the critical region to 1 Mb, we sequenced the gene Berardinelli-Seip congenital lipodystrophy (BSCL2) and identified two heterozygous missense mutations resulting in the amino acid substitutions N88S and S90L. 14981520 3 75 theme dHMN 632:635 arg1 syndrome 605:612 Silver syndrome 598:612 Silver syndrome 598:612 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 3 75 theme dHMN 632:635 arg1 forms 623:627 most forms 618:627 most forms of dHMN 618:635 Silver syndrome and most forms of dHMN are autosomal dominantly inherited with incomplete penetrance and a broad variability in clinical expression. 14981520 4 76 dep dHMN-V 793:798 arg1 ref 801:803 ref 801:803 ref 801:803 A genome-wide scan in an Austrian family with dHMN-V (ref. 14981520 1 77 theme heterogeneous 213:225 arg1 neuropathy 141:150 Distal hereditary motor neuropathy 117:150 Distal hereditary motor neuropathy (dHMN) 117:157 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 77 theme heterogeneous 213:225 arg1 disorders 236:244 disorders 236:244 disorders characterized by an almost exclusive degeneration of motor nerve fibers 236:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 77 theme heterogeneous 213:225 arg1 group 227:231 a heterogeneous group 211:231 a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers 211:316 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 14981520 1 77 theme heterogeneous 213:225 arg1 atrophy 185:191 distal spinal muscular atrophy 162:191 distal spinal muscular atrophy (OMIM #182960) 162:206 Distal hereditary motor neuropathy (dHMN) or distal spinal muscular atrophy (OMIM #182960) is a heterogeneous group of disorders characterized by an almost exclusive degeneration of motor nerve fibers, predominantly in the distal part of the limbs. 22020283 0 0 theme large 169:173 arg1 Members 97:103 Members 97:103 Members of the killer cell immunoglobulin-like receptor (KIR) family 97:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 0 theme large 169:173 arg1 group 175:179 a large group 167:179 a large group of polymorphic receptors expressed on natural killer (NK) cells 167:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 0 theme large 169:173 arg1 receptors 196:204 polymorphic receptors 184:204 polymorphic receptors expressed on natural killer (NK) cells 184:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 5 1 theme HLA 1348:1350 arg1 molecule 1352:1359 the HLA molecule 1344:1359 the HLA molecule 1344:1359 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 6 2 theme two-domain 1375:1384 arg1 KIR 1386:1388 the two-domain KIR 1371:1388 the two-domain KIR (KIR2D) 1371:1396 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 6 2 theme two-domain 1375:1384 arg1 KIR2D 1391:1395 KIR2D 1391:1395 KIR2D 1391:1395 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 5 3 theme D2-HLA-B 1130:1137 arg1 interface 1144:1152 the D2-HLA-B*5701 interface 1126:1152 the D2-HLA-B*5701 interface 1126:1152 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 1 4 dep domains 470:476 arg1 D0-D1-D2 478:485 D0-D1-D2 478:485 domains D0-D1-D2 470:485 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 2 5 theme HLA-B 705:709 arg1 * 710:710 HLA-B*5701 705:714 HLA-B*5701 complexed with a self-peptide 705:744 Here we describe the structure of a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide. 22020283 0 6 theme receptors 196:204 arg1 Members 97:103 Members 97:103 Members of the killer cell immunoglobulin-like receptor (KIR) family 97:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 6 theme receptors 196:204 arg1 group 175:179 a large group 167:179 a large group of polymorphic receptors expressed on natural killer (NK) cells 167:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 6 theme receptors 196:204 arg1 receptors 196:204 polymorphic receptors 184:204 polymorphic receptors expressed on natural killer (NK) cells 184:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 1 7 theme KIR 451:453 arg1 family 455:460 the three-domain KIR family 434:460 the three-domain KIR family (KIR3D, domains D0-D1-D2) 434:486 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 7 theme KIR 451:453 arg1 KIR3D 463:467 KIR3D 463:467 KIR3D 463:467 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 0 8 theme pivotal 341:347 arg1 role 349:352 a pivotal role 339:352 a pivotal role 339:352 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 9 theme primate 1951:1957 arg1 species 1959:1965 primate species 1951:1965 primate species 1951:1965 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 7 10 theme * 1707:1707 arg1 interface 1713:1721 the KIR3DL1-pHLA-B*5701 interface 1689:1721 the KIR3DL1-pHLA-B*5701 interface 1689:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 0 11 theme immunoglobulin-like 124:142 arg1 family 159:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 12 theme human 281:285 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 2 13 theme KIR3DL1 679:685 arg1 receptor 687:694 a human KIR3DL1 receptor 671:694 a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide 671:744 Here we describe the structure of a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide. 22020283 1 14 theme Allelic 382:388 arg1 variation 390:398 Allelic variation 382:398 Allelic variation 382:398 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 0 15 theme antigen 297:303 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 3 16 theme cleft 829:833 arg1 end 791:793 the carboxy-terminal end 770:793 the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft 770:833 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 3 17 from footprints 867:876 arg1 pHLA 885:888 the pHLA 881:888 the pHLA 881:888 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 3 18 theme discontinuous 853:865 arg1 footprints 867:876 two discontinuous footprints 849:876 two discontinuous footprints on the pHLA 849:888 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 5 19 theme D1-pHLA-B 1202:1210 arg1 suboptimal 1231:1240 suboptimal 1231:1240 suboptimal 1231:1240 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 5 19 theme D1-pHLA-B 1202:1210 arg1 contacts 1217:1224 the D1-pHLA-B*5701 contacts 1198:1224 the D1-pHLA-B*5701 contacts 1198:1224 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 3 20 theme carboxy-terminal 774:789 arg1 end 791:793 the carboxy-terminal end 770:793 the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft 770:833 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 0 21 theme leukocyte 287:295 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 22 theme pHLA 306:309 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 3 23 theme 5701 808:811 arg1 cleft 829:833 the HLA-B*5701 antigen-binding cleft 798:833 the HLA-B*5701 antigen-binding cleft 798:833 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 6 24 theme discrete 1586:1593 arg1 allotypes 1611:1619 discrete HLA-A and HLA-B allotypes 1586:1619 discrete HLA-A and HLA-B allotypes 1586:1619 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 4 25 theme HLA 1090:1092 arg1 domain 1102:1107 an 'innate HLA sensor' domain 1079:1107 an 'innate HLA sensor' domain 1079:1107 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 1 26 theme treatment 579:587 arg1 outcome 589:595 the treatment outcome 575:595 the treatment outcome of certain haematological malignancies 575:634 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 0 27 theme leukocyte 76:84 arg1 antigen 86:92 human leukocyte antigen 70:92 human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 70:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 28 theme class 312:316 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 29 with association 1639:1649 arg1 studies 1678:1684 extensive mutagenesis studies 1656:1684 extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface 1656:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 3 30 theme HLA-B 802:806 arg1 cleft 829:833 the HLA-B*5701 antigen-binding cleft 798:833 the HLA-B*5701 antigen-binding cleft 798:833 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 7 31 theme interaction 1914:1924 arg1 requirements 1876:1887 the specificity requirements 1860:1887 the specificity requirements of this essential innate interaction that is conserved across primate species 1860:1965 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 1 32 theme haematological 608:621 arg1 malignancies 623:634 certain haematological malignancies 600:634 certain haematological malignancies 600:634 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 7 33 theme essential 1897:1905 arg1 interaction 1914:1924 this essential innate interaction 1892:1924 this essential innate interaction that is conserved across primate species 1892:1965 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 7 34 theme mutagenesis 1666:1676 arg1 studies 1678:1684 extensive mutagenesis studies 1656:1684 extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface 1656:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 0 35 theme killer 112:117 arg1 family 159:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 36 theme Killer 0:5 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 37 theme specificity 1864:1874 arg1 requirements 1876:1887 the specificity requirements 1860:1887 the specificity requirements of this essential innate interaction that is conserved across primate species 1860:1965 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 0 38 theme particular 256:265 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 39 theme immunoglobulin-like 12:30 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 4 40 theme family 951:956 arg1 feature 930:936 a distinguishing feature 913:936 a distinguishing feature of the KIR3D family 913:956 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 4 40 theme family 951:956 arg1 domain 905:910 the D0 domain 898:910 the D0 domain 898:910 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 1 41 theme viral 553:557 arg1 replication 559:569 viral replication 553:569 viral replication 553:569 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 0 42 theme 3DL1-mediated 41:53 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 43 theme intricate 1769:1777 arg1 interplay 1779:1787 the intricate interplay 1765:1787 the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species 1765:1965 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 4 44 theme molecule 1025:1032 arg1 region 1007:1012 a region 1005:1012 a region of the HLA molecule with limited polymorphism 1005:1058 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 5 45 theme polymorphic 1322:1332 arg1 region 1334:1339 the polymorphic region 1318:1339 the polymorphic region of the HLA molecule 1318:1359 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 7 46 theme extensive 1656:1664 arg1 studies 1678:1684 extensive mutagenesis studies 1656:1684 extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface 1656:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 0 47 theme innate 357:362 arg1 responses 371:379 innate immune responses 357:379 innate immune responses 357:379 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 48 from interface 1713:1721 arg1 studies 1678:1684 extensive mutagenesis studies 1656:1684 extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface 1656:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 4 49 theme limited 1039:1045 arg1 polymorphism 1047:1058 limited polymorphism 1039:1058 limited polymorphism 1039:1058 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 5 50 theme variation 1280:1288 arg1 degree 1261:1266 a degree 1259:1266 a degree of sequence variation 1259:1288 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 0 51 theme polymorphic 184:194 arg1 receptors 196:204 polymorphic receptors 184:204 polymorphic receptors expressed on natural killer (NK) cells 184:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 5 52 theme molecule 1352:1359 arg1 peptide 1306:1312 the peptide 1302:1312 the peptide 1302:1312 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 5 52 theme molecule 1352:1359 arg1 region 1334:1339 the polymorphic region 1318:1339 the polymorphic region of the HLA molecule 1318:1359 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 5 53 theme * 1138:1138 arg1 interface 1144:1152 the D2-HLA-B*5701 interface 1126:1152 the D2-HLA-B*5701 interface 1126:1152 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 1 54 theme three-domain 438:449 arg1 family 455:460 the three-domain KIR family 434:460 the three-domain KIR family (KIR3D, domains D0-D1-D2) 434:486 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 54 theme three-domain 438:449 arg1 KIR3D 463:467 KIR3D 463:467 KIR3D 463:467 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 7 55 theme 5701 1708:1711 arg1 interface 1713:1721 the KIR3DL1-pHLA-B*5701 interface 1689:1721 the KIR3DL1-pHLA-B*5701 interface 1689:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 4 56 theme innate 1083:1088 arg1 domain 1102:1107 an 'innate HLA sensor' domain 1079:1107 an 'innate HLA sensor' domain 1079:1107 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 1 57 theme malignancies 623:634 arg1 control 542:548 the control 538:548 the control of viral replication 538:569 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 57 theme malignancies 623:634 arg1 outcome 589:595 the treatment outcome 575:595 the treatment outcome of certain haematological malignancies 575:634 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 0 58 theme natural 219:225 arg1 cells 239:243 natural killer (NK) cells 219:243 natural killer (NK) cells 219:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 59 theme KIR3DL1-pHLA-B 1693:1706 arg1 interface 1713:1721 the KIR3DL1-pHLA-B*5701 interface 1689:1721 the KIR3DL1-pHLA-B*5701 interface 1689:1721 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 0 60 theme receptor 144:151 arg1 family 159:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 61 theme KIR 154:156 arg1 family 159:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 5 62 theme complementarity 1181:1195 arg1 degree 1171:1176 a high degree 1164:1176 a high degree of complementarity 1164:1195 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 1 63 attach linked 528:533 arg1 control 542:548 the control 538:548 the control of viral replication 538:569 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 63 attach linked 528:533 arg2 variation 390:398 Allelic variation 382:398 Allelic variation 382:398 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 63 attach linked 528:533 arg1 outcome 589:595 the treatment outcome 575:595 the treatment outcome of certain haematological malignancies 575:634 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 2 64 theme receptor 687:694 arg1 structure 658:666 the structure 654:666 the structure of a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide 654:744 Here we describe the structure of a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide. 22020283 0 65 theme family 159:164 arg1 Members 97:103 Members 97:103 Members of the killer cell immunoglobulin-like receptor (KIR) family 97:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 65 theme family 159:164 arg1 group 175:179 a large group 167:179 a large group of polymorphic receptors expressed on natural killer (NK) cells 167:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 65 theme family 159:164 arg1 receptors 196:204 polymorphic receptors 184:204 polymorphic receptors expressed on natural killer (NK) cells 184:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 66 theme peptide-laden 267:279 arg1 molecules 320:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 particular peptide-laden human leukocyte antigen (pHLA) class I molecules 256:328 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 6 67 theme HLA-B 1605:1609 arg1 allotypes 1611:1619 discrete HLA-A and HLA-B allotypes 1586:1619 discrete HLA-A and HLA-B allotypes 1586:1619 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 2 68 theme human 673:677 arg1 receptor 687:694 a human KIR3DL1 receptor 671:694 a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide 671:744 Here we describe the structure of a human KIR3DL1 receptor bound to HLA-B*5701 complexed with a self-peptide. 22020283 1 69 theme replication 559:569 arg1 control 542:548 the control 538:548 the control of viral replication 538:569 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 69 theme replication 559:569 arg1 outcome 589:595 the treatment outcome 575:595 the treatment outcome of certain haematological malignancies 575:634 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 6 70 theme KIR3DL1 1574:1580 arg1 specificity 1559:1569 the specificity 1555:1569 the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes 1555:1619 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 6 71 theme HLA-A 1595:1599 arg1 allotypes 1611:1619 discrete HLA-A and HLA-B allotypes 1586:1619 discrete HLA-A and HLA-B allotypes 1586:1619 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 4 72 theme sensor 1094:1099 arg1 domain 1102:1107 an 'innate HLA sensor' domain 1079:1107 an 'innate HLA sensor' domain 1079:1107 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 5 73 theme * 1211:1211 arg1 suboptimal 1231:1240 suboptimal 1231:1240 suboptimal 1231:1240 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 5 73 theme * 1211:1211 arg1 contacts 1217:1224 the D1-pHLA-B*5701 contacts 1198:1224 the D1-pHLA-B*5701 contacts 1198:1224 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 0 74 theme antigen 86:92 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 1 75 theme extensive 404:412 arg1 polymorphism 414:425 extensive polymorphism 404:425 extensive polymorphism 404:425 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 6 76 theme D1-mediated 1480:1490 arg1 interactions 1492:1503 the corresponding D1-mediated interactions 1462:1503 the corresponding D1-mediated interactions 1462:1503 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 3 77 theme antigen-binding 813:827 arg1 cleft 829:833 the HLA-B*5701 antigen-binding cleft 798:833 the HLA-B*5701 antigen-binding cleft 798:833 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 4 78 theme distinguishing 915:928 arg1 feature 930:936 a distinguishing feature 913:936 a distinguishing feature of the KIR3D family 913:956 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 4 78 theme distinguishing 915:928 arg1 domain 905:910 the D0 domain 898:910 the D0 domain 898:910 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 7 79 theme innate 1907:1912 arg1 interaction 1914:1924 this essential innate interaction 1892:1924 this essential innate interaction that is conserved across primate species 1892:1965 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 1 80 dep KIR3D 463:467 arg1 domains 470:476 domains D0-D1-D2 470:485 domains D0-D1-D2 470:485 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 1 81 theme certain 600:606 arg1 malignancies 623:634 certain haematological malignancies 600:634 certain haematological malignancies 600:634 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 3 82 theme * 807:807 arg1 cleft 829:833 the HLA-B*5701 antigen-binding cleft 798:833 the HLA-B*5701 antigen-binding cleft 798:833 KIR3DL1 clamped around the carboxy-terminal end of the HLA-B*5701 antigen-binding cleft, resulting in two discontinuous footprints on the pHLA. 22020283 0 83 theme killer 227:232 arg1 cells 239:243 natural killer (NK) cells 219:243 natural killer (NK) cells 219:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 4 84 theme D0 902:903 arg1 feature 930:936 a distinguishing feature 913:936 a distinguishing feature of the KIR3D family 913:956 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 4 84 theme D0 902:903 arg1 domain 905:910 the D0 domain 898:910 the D0 domain 898:910 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 0 85 theme cell 119:122 arg1 family 159:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 the killer cell immunoglobulin-like receptor (KIR) family 108:164 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 86 theme NK 235:236 arg1 cells 239:243 natural killer (NK) cells 219:243 natural killer (NK) cells 219:243 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 87 theme cell 7:10 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 4 88 with region 1007:1012 arg1 polymorphism 1047:1058 limited polymorphism 1039:1058 limited polymorphism 1039:1058 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 0 89 dep Members 97:103 arg1 B. 94:95 B. 94:95 B. 94:95 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 5 90 dep peptide 1306:1312 arg1 both 1290:1293 both 1290:1293 both 1290:1293 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 0 91 theme receptor 32:39 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 1 92 theme pHLA 496:499 arg1 specificity 509:519 pHLA binding specificity 496:519 pHLA binding specificity 496:519 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 7 93 theme HLA 1828:1830 arg1 polymorphism 1832:1843 HLA polymorphism 1828:1843 HLA polymorphism 1828:1843 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 4 94 theme KIR3D 945:949 arg1 family 951:956 the KIR3D family 941:956 the KIR3D family 941:956 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 6 95 theme corresponding 1466:1478 arg1 interactions 1492:1503 the corresponding D1-mediated interactions 1462:1503 the corresponding D1-mediated interactions 1462:1503 Although the two-domain KIR (KIR2D) and KIR3DL1 docked similarly onto HLA-C and HLA-B respectively, the corresponding D1-mediated interactions differed markedly, thereby providing insight into the specificity of KIR3DL1 for discrete HLA-A and HLA-B allotypes. 22020283 0 96 theme human 70:74 arg1 antigen 86:92 human leukocyte antigen 70:92 human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 70:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 7 97 theme peptide 1797:1803 arg1 variability 1805:1815 peptide variability 1797:1815 peptide variability 1797:1815 Collectively, in association with extensive mutagenesis studies at the KIR3DL1-pHLA-B*5701 interface, we provide a framework for understanding the intricate interplay between peptide variability, KIR3D and HLA polymorphism in determining the specificity requirements of this essential innate interaction that is conserved across primate species. 22020283 0 98 contain have 334:337 arg1 recognition 55:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition 0:65 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, 0:244 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 98 contain have 334:337 arg2 role 349:352 a pivotal role 339:352 a pivotal role 339:352 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 0 99 theme immune 364:369 arg1 responses 371:379 innate immune responses 357:379 innate immune responses 357:379 Killer cell immunoglobulin-like receptor 3DL1-mediated recognition of human leukocyte antigen B. Members of the killer cell immunoglobulin-like receptor (KIR) family, a large group of polymorphic receptors expressed on natural killer (NK) cells, recognize particular peptide-laden human leukocyte antigen (pHLA) class I molecules and have a pivotal role in innate immune responses. 22020283 4 100 theme HLA 1021:1023 arg1 molecule 1025:1032 the HLA molecule 1017:1032 the HLA molecule 1017:1032 First, the D0 domain, a distinguishing feature of the KIR3D family, extended towards β2-microglobulin and abutted a region of the HLA molecule with limited polymorphism, thereby acting as an 'innate HLA sensor' domain. 22020283 5 101 theme sequence 1271:1278 arg1 variation 1280:1288 sequence variation 1271:1288 sequence variation 1271:1288 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 22020283 1 102 theme binding 501:507 arg1 specificity 509:519 pHLA binding specificity 496:519 pHLA binding specificity 496:519 Allelic variation and extensive polymorphism within the three-domain KIR family (KIR3D, domains D0-D1-D2) affects pHLA binding specificity and is linked to the control of viral replication and the treatment outcome of certain haematological malignancies. 22020283 5 103 theme high 1166:1169 arg1 degree 1171:1176 a high degree 1164:1176 a high degree of complementarity 1164:1195 Second, whereas the D2-HLA-B*5701 interface exhibited a high degree of complementarity, the D1-pHLA-B*5701 contacts were suboptimal and accommodated a degree of sequence variation both within the peptide and the polymorphic region of the HLA molecule. 29490423 3 0 with complex 580:586 arg1 CD19 593:596 CD19 593:596 CD19 593:596 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 1 1 dep monoclonal 250:259 arg1 antibody-mediated 261:277 antibody-mediated 261:277 antibody-mediated 261:277 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 4 2 theme binding 650:656 arg1 epitope 658:664 the binding epitope 646:664 the binding epitope 646:664 The structure revealed the binding epitope, explained the lack of cross-reactivity toward non-human species, and suggested the key-and-lock mechanism of antigen recognition. 29490423 6 3 dep Rather 876:881 arg1 folds 922:926 folds 922:926 folds predicted from the amino acid sequence 922:965 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 0 4 from structure 8:16 arg1 complex 48:54 complex 48:54 complex with antibody B43 48:72 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 1 5 theme malignant 147:155 arg1 cells 159:163 malignant B cells 147:163 malignant B cells 147:163 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 0 6 theme unexpected 85:94 arg1 fold 96:99 an unexpected fold 82:99 an unexpected fold 82:99 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 6 7 theme acid 953:956 arg1 sequence 958:965 the amino acid sequence 943:965 the amino acid sequence 943:965 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 3 8 theme crystal 533:539 arg1 structure 541:549 the crystal structure 529:549 the crystal structure of B43 Fab 529:560 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 4 9 theme non-human 713:721 arg1 species 723:729 non-human species 713:729 non-human species 713:729 The structure revealed the binding epitope, explained the lack of cross-reactivity toward non-human species, and suggested the key-and-lock mechanism of antigen recognition. 29490423 3 10 theme antibody 519:526 arg1 action 505:510 action 505:510 action of the antibody 505:526 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 2 11 theme antibody 304:311 arg1 B43 313:315 Anti-CD19 antibody B43 294:315 Anti-CD19 antibody B43 294:315 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 3 12 theme action 505:510 arg1 mechanism 492:500 the mechanism 488:500 the mechanism of action of the antibody 488:526 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 1 13 theme B 157:157 arg1 cells 159:163 malignant B cells 147:163 malignant B cells 147:163 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 6 14 theme amino 947:951 arg1 sequence 958:965 the amino acid sequence 943:965 the amino acid sequence 943:965 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 5 15 theme CD19 870:873 arg1 topology 858:865 a unique molecular topology 839:865 a unique molecular topology of CD19 839:873 Most unexpectedly, the structure revealed a unique molecular topology of CD19. 29490423 2 16 theme leukemia 457:464 arg1 treatment 415:423 the treatment 411:423 the treatment of relapsed acute lymphoblastic leukemia 411:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 2 17 theme BiTE 362:365 arg1 blinatumomab 368:379 a bispecific T-cell engager (BiTE) blinatumomab 333:379 a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia 333:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 7 18 theme CD19 1137:1140 arg1 This 1106:1109 This 1106:1109 This 1106:1109 This is the first structure of CD19, which has no sequence homologs. 29490423 7 18 theme CD19 1137:1140 arg1 structure 1124:1132 the first structure 1114:1132 the first structure 1114:1132 This is the first structure of CD19, which has no sequence homologs. 29490423 2 19 theme Anti-CD19 294:302 arg1 B43 313:315 Anti-CD19 antibody B43 294:315 Anti-CD19 antibody B43 294:315 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 0 20 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 0:72 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 1 21 theme attractive 228:237 arg1 target 239:244 an attractive target 225:244 an attractive target for monoclonal antibody-mediated immunotherapy 225:291 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 2 22 theme engager 353:359 arg1 blinatumomab 368:379 a bispecific T-cell engager (BiTE) blinatumomab 333:379 a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia 333:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 0 23 theme B-cell 21:26 arg1 CD19 40:43 B-cell co-receptor CD19 21:43 B-cell co-receptor CD19 21:43 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 2 24 used utilized 321:328 arg2 B43 313:315 Anti-CD19 antibody B43 294:315 Anti-CD19 antibody B43 294:315 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 4 25 theme antigen 776:782 arg1 recognition 784:794 antigen recognition 776:794 antigen recognition 776:794 The structure revealed the binding epitope, explained the lack of cross-reactivity toward non-human species, and suggested the key-and-lock mechanism of antigen recognition. 29490423 6 26 theme elongated 1013:1021 arg1 β-sandwich 1023:1032 an elongated β-sandwich 1010:1032 an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves 1010:1103 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 4 27 theme cross-reactivity 689:704 arg1 lack 681:684 the lack 677:684 the lack of cross-reactivity toward non-human species 677:729 The structure revealed the binding epitope, explained the lack of cross-reactivity toward non-human species, and suggested the key-and-lock mechanism of antigen recognition. 29490423 7 28 theme first 1118:1122 arg1 This 1106:1109 This 1106:1109 This 1106:1109 This is the first structure of CD19, which has no sequence homologs. 29490423 7 28 theme first 1118:1122 arg1 structure 1124:1132 the first structure 1114:1132 the first structure 1114:1132 This is the first structure of CD19, which has no sequence homologs. 29490423 5 29 theme molecular 848:856 arg1 topology 858:865 a unique molecular topology 839:865 a unique molecular topology of CD19 839:873 Most unexpectedly, the structure revealed a unique molecular topology of CD19. 29490423 0 30 theme CD19 40:43 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 0:72 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 1 31 theme monoclonal 250:259 arg1 immunotherapy 279:291 monoclonal antibody-mediated immunotherapy 250:291 monoclonal antibody-mediated immunotherapy 250:291 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 3 32 theme Fab 558:560 arg1 structure 541:549 the crystal structure 529:549 the crystal structure of B43 Fab 529:560 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 5 33 theme unique 841:846 arg1 topology 858:865 a unique molecular topology 839:865 a unique molecular topology of CD19 839:873 Most unexpectedly, the structure revealed a unique molecular topology of CD19. 29490423 1 34 theme other 177:181 arg1 lineages 183:190 other lineages 177:190 other lineages 177:190 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 0 35 theme co-receptor 28:38 arg1 CD19 40:43 B-cell co-receptor CD19 21:43 B-cell co-receptor CD19 21:43 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 6 36 theme immunoglobulin 907:920 arg1 tandem 890:895 tandem 890:895 tandem 890:895 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 2 37 theme lymphoblastic 443:455 arg1 leukemia 457:464 relapsed acute lymphoblastic leukemia 428:464 relapsed acute lymphoblastic leukemia 428:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 6 38 dep immunoglobulin 1048:1061 arg1 folds 1063:1067 folds 1063:1067 folds 1063:1067 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 7 39 contain has 1149:1151 arg1 This 1106:1109 This 1106:1109 This 1106:1109 This is the first structure of CD19, which has no sequence homologs. 29490423 7 39 contain has 1149:1151 arg2 homologs 1165:1172 no sequence homologs 1153:1172 no sequence homologs 1153:1172 This is the first structure of CD19, which has no sequence homologs. 29490423 7 39 contain has 1149:1151 arg1 structure 1124:1132 the first structure 1114:1132 the first structure 1114:1132 This is the first structure of CD19, which has no sequence homologs. 29490423 6 40 theme CD19 996:999 arg1 CD19 996:999 CD19 996:999 CD19 996:999 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 6 40 theme CD19 996:999 arg1 domain 986:991 the extracellular domain 968:991 the extracellular domain of CD19 968:999 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 2 41 theme acute 437:441 arg1 leukemia 457:464 relapsed acute lymphoblastic leukemia 428:464 relapsed acute lymphoblastic leukemia 428:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 6 42 theme c-type 900:905 arg1 immunoglobulin 907:920 c-type immunoglobulin 900:920 c-type immunoglobulin 900:920 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 4 43 theme recognition 784:794 arg1 mechanism 763:771 the key-and-lock mechanism 746:771 the key-and-lock mechanism of antigen recognition 746:794 The structure revealed the binding epitope, explained the lack of cross-reactivity toward non-human species, and suggested the key-and-lock mechanism of antigen recognition. 29490423 2 44 theme relapsed 428:435 arg1 leukemia 457:464 relapsed acute lymphoblastic leukemia 428:464 relapsed acute lymphoblastic leukemia 428:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 3 45 theme B43 554:556 arg1 Fab 558:560 B43 Fab 554:560 B43 Fab 554:560 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 6 46 dep folds 922:926 arg1 predicted 928:936 predicted 928:936 folds predicted from the amino acid sequence 922:965 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 6 47 theme C-terminal 1087:1096 arg1 halves 1098:1103 their C-terminal halves 1081:1103 their C-terminal halves 1081:1103 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 3 48 theme unbound 609:615 arg1 form 617:620 the unbound form 605:620 the unbound form 605:620 To gain insight into the mechanism of action of the antibody, the crystal structure of B43 Fab was determined in complex with CD19 and in the unbound form. 29490423 0 49 with complex 48:54 arg1 antibody 61:68 antibody B43 61:72 antibody B43 61:72 Crystal structure of B-cell co-receptor CD19 in complex with antibody B43 reveals an unexpected fold. 29490423 1 50 theme transmembrane 112:124 arg1 CD19 102:105 CD19 102:105 CD19 102:105 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 1 50 theme transmembrane 112:124 arg1 protein 126:132 a transmembrane protein 110:132 a transmembrane protein 110:132 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 1 51 theme other 195:199 arg1 tissues 201:207 other tissues 195:207 other tissues 195:207 CD19 is a transmembrane protein expressed on malignant B cells, but not in other lineages or other tissues, which makes it an attractive target for monoclonal antibody-mediated immunotherapy. 29490423 2 52 theme T-cell 346:351 arg1 blinatumomab 368:379 a bispecific T-cell engager (BiTE) blinatumomab 333:379 a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia 333:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 6 53 theme extracellular 972:984 arg1 CD19 996:999 CD19 996:999 CD19 996:999 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 6 53 theme extracellular 972:984 arg1 domain 986:991 the extracellular domain 968:991 the extracellular domain of CD19 968:999 Rather than a tandem of c-type immunoglobulin folds predicted from the amino acid sequence, the extracellular domain of CD19 exhibits an elongated β-sandwich formed by two immunoglobulin folds by swapping their C-terminal halves. 29490423 4 54 theme key-and-lock 750:761 arg1 mechanism 763:771 the key-and-lock mechanism 746:771 the key-and-lock mechanism of antigen recognition 746:794 The structure revealed the binding epitope, explained the lack of cross-reactivity toward non-human species, and suggested the key-and-lock mechanism of antigen recognition. 29490423 2 55 theme bispecific 335:344 arg1 blinatumomab 368:379 a bispecific T-cell engager (BiTE) blinatumomab 333:379 a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia 333:464 Anti-CD19 antibody B43 was utilized in a bispecific T-cell engager (BiTE) blinatumomab that demonstrated potency for the treatment of relapsed acute lymphoblastic leukemia. 29490423 7 56 theme sequence 1156:1163 arg1 homologs 1165:1172 no sequence homologs 1153:1172 no sequence homologs 1153:1172 This is the first structure of CD19, which has no sequence homologs. 12034737 0 0 theme stromal-derived 76:90 arg1 association 107:117 stromal-derived factor 1 alpha association 76:117 stromal-derived factor 1 alpha association 76:117 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 5 1 mod modified 724:731 arg1 CXCR4 703:707 CXCR4 703:707 CXCR4 703:707 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 5 1 mod modified 724:731 arg3 chain 758:762 a chondroitin sulfate chain 736:762 a chondroitin sulfate chain at serine 18 736:775 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 1 2 theme receptor 150:157 arg1 CXCR4 159:163 The chemokine receptor CXCR4 136:163 The chemokine receptor CXCR4 136:163 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 1 3 theme virus 246:250 arg1 type 252:255 human immunodeficiency virus type 1 223:257 human immunodeficiency virus type 1 (HIV-1) entry 223:271 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 1 3 theme virus 246:250 arg1 HIV-1 260:264 HIV-1 260:264 HIV-1 260:264 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 3 4 link derived 543:549 arg1 alpha 560:564 stromal derived factor 1 alpha 535:564 stromal derived factor 1 alpha 535:564 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 3 4 link derived 543:549 arg1 ligand 527:532 its ligand 523:532 its ligand 523:532 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 0 5 from role 4:7 arg1 entry 129:133 HIV-1 entry 123:133 HIV-1 entry 123:133 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 0 5 from role 4:7 arg1 association 107:117 stromal-derived factor 1 alpha association 76:117 stromal-derived factor 1 alpha association 76:117 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 4 6 theme HIV-1 670:674 arg1 entry 676:680 CCR5-dependent HIV-1 entry 655:680 CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry 619:680 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 1 7 theme type 252:255 arg1 entry 267:271 human immunodeficiency virus type 1 (HIV-1) entry 223:271 human immunodeficiency virus type 1 (HIV-1) entry 223:271 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 6 8 theme CCR5 1052:1055 arg1 utilization 1037:1047 HIV-1 utilization 1031:1047 HIV-1 utilization of CCR5 and CXCR4 1031:1065 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 6 9 from difference 1017:1026 arg1 utilization 1037:1047 HIV-1 utilization 1031:1047 HIV-1 utilization of CCR5 and CXCR4 1031:1065 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 6 10 theme tyrosine 938:945 arg1 sulfate 947:953 tyrosine sulfate 938:953 tyrosine sulfate 938:953 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 0 11 theme factor 92:97 arg1 association 107:117 stromal-derived factor 1 alpha association 76:117 stromal-derived factor 1 alpha association 76:117 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 2 12 dep receptors 322:330 arg1 CCR5 332:335 CCR5 332:335 CCR5 332:335 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 2 12 dep receptors 322:330 arg1 receptors 322:330 the CC-chemokine receptors CCR5 and CCR2b 305:345 the CC-chemokine receptors CCR5 and CCR2b 305:345 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 2 12 dep receptors 322:330 arg1 CCR2b 341:345 CCR2b 341:345 CCR2b 341:345 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 5 13 theme stromal 806:812 arg1 factor 822:827 stromal derived factor 1 806:829 stromal derived factor 1 alpha binding 806:843 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 5 14 theme derived 814:820 arg1 factor 822:827 stromal derived factor 1 806:829 stromal derived factor 1 alpha binding 806:843 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 3 15 theme CXCR4 509:513 arg1 ability 498:504 the ability 494:504 the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha 494:564 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 4 16 theme HIV-1 635:639 arg1 entry 641:645 CXCR4-dependent HIV-1 entry 619:645 CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry 619:680 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 0 17 theme alpha 101:105 arg1 association 107:117 stromal-derived factor 1 alpha association 76:117 stromal-derived factor 1 alpha association 76:117 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 5 18 from serine 767:772 arg1 chain 758:762 a chondroitin sulfate chain 736:762 a chondroitin sulfate chain at serine 18 736:775 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 3 19 from tyrosine 453:460 arg1 group 444:448 The sulfate group 432:448 The sulfate group at tyrosine 21 432:463 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 5 20 theme glycosaminoglycan 874:890 arg1 loss 861:864 loss 861:864 loss of this glycosaminoglycan 861:890 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 1 21 theme critical 171:178 arg1 roles 180:184 critical roles 171:184 critical roles 171:184 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 3 22 theme stromal 535:541 arg1 alpha 560:564 stromal derived factor 1 alpha 535:564 stromal derived factor 1 alpha 535:564 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 3 22 theme stromal 535:541 arg1 ligand 527:532 its ligand 523:532 its ligand 523:532 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 5 23 theme factor 822:827 arg1 binding 837:843 stromal derived factor 1 alpha binding 806:843 stromal derived factor 1 alpha binding 806:843 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 2 24 theme tyrosines 421:429 arg1 sulfation 389:397 sulfation 389:397 sulfation of its amino-terminal tyrosines 389:429 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 5 25 link derived 814:820 arg1 factor 822:827 stromal derived factor 1 806:829 stromal derived factor 1 alpha binding 806:843 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 3 26 theme derived 543:549 arg1 alpha 560:564 stromal derived factor 1 alpha 535:564 stromal derived factor 1 alpha 535:564 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 3 26 theme derived 543:549 arg1 ligand 527:532 its ligand 523:532 its ligand 523:532 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 2 27 mod modified 377:384 arg1 CXCR4 348:352 CXCR4 348:352 CXCR4 348:352 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 2 27 mod modified 377:384 arg3 sulfation 389:397 sulfation 389:397 sulfation of its amino-terminal tyrosines 389:429 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 0 28 theme post-translational 12:29 arg1 modifications 31:43 post-translational modifications 12:43 post-translational modifications of the CXCR4 amino terminus 12:71 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 6 29 theme HIV-1 1031:1035 arg1 utilization 1037:1047 HIV-1 utilization 1031:1047 HIV-1 utilization of CCR5 and CXCR4 1031:1065 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 0 30 theme HIV-1 123:127 arg1 entry 129:133 HIV-1 entry 123:133 HIV-1 entry 123:133 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 4 31 theme significant 599:609 arg1 role 611:614 a less significant role 592:614 a less significant role 592:614 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 6 32 theme CXC-chemokine 962:974 arg1 family 985:990 the CXC-chemokine receptor family 958:990 the CXC-chemokine receptor family 958:990 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 5 33 theme HIV-1 790:794 arg1 entry 796:800 HIV-1 entry 790:800 HIV-1 entry 790:800 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 5 34 theme chondroitin 738:748 arg1 chain 758:762 a chondroitin sulfate chain 736:762 a chondroitin sulfate chain at serine 18 736:775 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 6 35 from role 929:932 arg1 family 985:990 the CXC-chemokine receptor family 958:990 the CXC-chemokine receptor family 958:990 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 1 36 theme immune 202:207 arg1 function 209:216 immune function 202:216 immune function 202:216 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 0 37 theme modifications 31:43 arg1 role 4:7 The role 0:7 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 0:134 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 5 38 theme cell 691:694 arg1 lines 696:700 some cell lines 686:700 some cell lines 686:700 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 2 39 theme amino-terminal 406:419 arg1 tyrosines 421:429 its amino-terminal tyrosines 402:429 its amino-terminal tyrosines 402:429 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 0 40 link stromal-derived 76:90 arg1 association 107:117 stromal-derived factor 1 alpha association 76:117 stromal-derived factor 1 alpha association 76:117 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 3 41 theme sulfate 436:442 arg1 group 444:448 The sulfate group 432:448 The sulfate group at tyrosine 21 432:463 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 0 42 theme CXCR4 52:56 arg1 terminus 64:71 the CXCR4 amino terminus 48:71 the CXCR4 amino terminus 48:71 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 5 43 theme alpha 831:835 arg1 binding 837:843 stromal derived factor 1 alpha binding 806:843 stromal derived factor 1 alpha binding 806:843 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 6 44 theme functional 918:927 arg1 role 929:932 a functional role 916:932 a functional role for tyrosine sulfate in the CXC-chemokine receptor family 916:990 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 3 45 theme factor 551:556 arg1 alpha 560:564 stromal derived factor 1 alpha 535:564 stromal derived factor 1 alpha 535:564 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 3 45 theme factor 551:556 arg1 ligand 527:532 its ligand 523:532 its ligand 523:532 The sulfate group at tyrosine 21 contributes substantially to the ability of CXCR4 to bind its ligand, stromal derived factor 1 alpha. 12034737 6 46 theme CXCR4 1061:1065 arg1 utilization 1037:1047 HIV-1 utilization 1031:1047 HIV-1 utilization of CCR5 and CXCR4 1031:1065 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 2 47 theme CC-chemokine 309:320 arg1 CCR5 332:335 CCR5 332:335 CCR5 332:335 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 2 47 theme CC-chemokine 309:320 arg1 receptors 322:330 the CC-chemokine receptors CCR5 and CCR2b 305:345 the CC-chemokine receptors CCR5 and CCR2b 305:345 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 2 47 theme CC-chemokine 309:320 arg1 CCR2b 341:345 CCR2b 341:345 CCR2b 341:345 Here we demonstrate that, like the CC-chemokine receptors CCR5 and CCR2b, CXCR4 is posttranslationally modified by sulfation of its amino-terminal tyrosines. 12034737 4 48 dep than 647:650 arg1 in 652:653 in 652:653 in 652:653 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 6 49 theme receptor 976:983 arg1 family 985:990 the CXC-chemokine receptor family 958:990 the CXC-chemokine receptor family 958:990 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 4 50 theme Tyrosine 567:574 arg1 sulfation 576:584 Tyrosine sulfation 567:584 Tyrosine sulfation 567:584 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 0 51 theme terminus 64:71 arg1 modifications 31:43 post-translational modifications 12:43 post-translational modifications of the CXCR4 amino terminus 12:71 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 6 52 theme general 1009:1015 arg1 difference 1017:1026 a general difference 1007:1026 a general difference in HIV-1 utilization of CCR5 and CXCR4 1007:1065 These data demonstrate a functional role for tyrosine sulfate in the CXC-chemokine receptor family and underscore a general difference in HIV-1 utilization of CCR5 and CXCR4. 12034737 4 53 theme CCR5-dependent 655:668 arg1 entry 676:680 CCR5-dependent HIV-1 entry 655:680 CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry 619:680 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 4 54 theme CXCR4-dependent 619:633 arg1 entry 641:645 CXCR4-dependent HIV-1 entry 619:645 CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry 619:680 Tyrosine sulfation plays a less significant role in CXCR4-dependent HIV-1 entry than in CCR5-dependent HIV-1 entry. 12034737 1 55 theme human 223:227 arg1 type 252:255 human immunodeficiency virus type 1 223:257 human immunodeficiency virus type 1 (HIV-1) entry 223:271 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 1 55 theme human 223:227 arg1 HIV-1 260:264 HIV-1 260:264 HIV-1 260:264 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 0 56 theme amino 58:62 arg1 terminus 64:71 the CXCR4 amino terminus 48:71 the CXCR4 amino terminus 48:71 The role of post-translational modifications of the CXCR4 amino terminus in stromal-derived factor 1 alpha association and HIV-1 entry. 12034737 5 57 theme sulfate 750:756 arg1 chain 758:762 a chondroitin sulfate chain 736:762 a chondroitin sulfate chain at serine 18 736:775 In some cell lines, CXCR4 is efficiently modified by a chondroitin sulfate chain at serine 18, but neither HIV-1 entry nor stromal derived factor 1 alpha binding was affected by loss of this glycosaminoglycan. 12034737 1 58 theme chemokine 140:148 arg1 CXCR4 159:163 The chemokine receptor CXCR4 136:163 The chemokine receptor CXCR4 136:163 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 1 59 theme immunodeficiency 229:244 arg1 type 252:255 human immunodeficiency virus type 1 223:257 human immunodeficiency virus type 1 (HIV-1) entry 223:271 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 12034737 1 59 theme immunodeficiency 229:244 arg1 HIV-1 260:264 HIV-1 260:264 HIV-1 260:264 The chemokine receptor CXCR4 plays critical roles in development, immune function, and human immunodeficiency virus type 1 (HIV-1) entry. 17113084 5 0 theme 72kDa 721:725 arg1 glycoform 727:735 the 72kDa glycoform 717:735 the 72kDa glycoform 717:735 Thus, COX-2 turnover appears to depend upon glycosylation of the 72kDa glycoform. 17113084 4 1 theme mutant 471:476 arg1 gene 478:481 the mutant gene 467:481 the mutant gene 467:481 COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. 17113084 2 2 theme additional 275:284 arg1 chain 302:306 an additional oligosaccharide chain 272:306 an additional oligosaccharide chain 272:306 COX-2 exists as 72 and 74kDa glycoforms, the latter resulting from an additional oligosaccharide chain at residue Asn(580). 17113084 2 3 theme residue 311:317 arg1 580 323:325 580 323:325 580 323:325 COX-2 exists as 72 and 74kDa glycoforms, the latter resulting from an additional oligosaccharide chain at residue Asn(580). 17113084 2 3 theme residue 311:317 arg1 Asn 319:321 residue Asn 311:321 residue Asn(580) 311:326 COX-2 exists as 72 and 74kDa glycoforms, the latter resulting from an additional oligosaccharide chain at residue Asn(580). 17113084 4 4 theme COS-1 438:442 arg1 cells 444:448 COS-1 cells 438:448 COS-1 cells transfected with the mutant gene 438:481 COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. 17113084 5 5 gly glycosylation 700:712 arg1 glycoform 727:735 the 72kDa glycoform 717:735 the 72kDa glycoform 717:735 Thus, COX-2 turnover appears to depend upon glycosylation of the 72kDa glycoform. 17113084 1 6 theme prostaglandin 185:197 arg1 2 201:201 2 201:201 2 201:201 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 17113084 1 6 theme prostaglandin 185:197 arg1 H 199:199 prostaglandin H 185:199 prostaglandin H(2) 185:202 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 17113084 4 7 theme COX-2 602:606 arg1 activity 608:615 five times greater COX-2 activity 583:615 five times greater COX-2 activity than cells expressing both glycoforms 583:653 COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. 17113084 1 8 theme rate-limiting 93:105 arg1 step 107:110 the rate-limiting step 89:110 the rate-limiting step in the prostanoid biosynthesis pathway 89:149 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 17113084 4 9 theme 74kDa 510:514 arg1 glycoform 516:524 the 74kDa glycoform 506:524 the 74kDa glycoform 506:524 COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. 17113084 3 10 theme variable 414:421 arg1 glycosylation 423:435 this variable glycosylation 409:435 this variable glycosylation 409:435 In this study, Asn(580) was mutated to determine the biological significance of this variable glycosylation. 17113084 2 11 theme oligosaccharide 286:300 arg1 chain 302:306 an additional oligosaccharide chain 272:306 an additional oligosaccharide chain 272:306 COX-2 exists as 72 and 74kDa glycoforms, the latter resulting from an additional oligosaccharide chain at residue Asn(580). 17113084 3 12 theme glycosylation 423:435 arg1 significance 393:404 the biological significance 378:404 the biological significance of this variable glycosylation 378:435 In this study, Asn(580) was mutated to determine the biological significance of this variable glycosylation. 17113084 5 13 theme glycoform 727:735 arg1 glycosylation 700:712 glycosylation 700:712 glycosylation of the 72kDa glycoform 700:735 Thus, COX-2 turnover appears to depend upon glycosylation of the 72kDa glycoform. 17113084 5 14 theme COX-2 662:666 arg1 turnover 668:675 COX-2 turnover 662:675 COX-2 turnover 662:675 Thus, COX-2 turnover appears to depend upon glycosylation of the 72kDa glycoform. 17113084 4 15 theme greater 594:600 arg1 activity 608:615 five times greater COX-2 activity 583:615 five times greater COX-2 activity than cells expressing both glycoforms 583:653 COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. 17113084 0 16 theme cyclooxygenase-2 36:51 arg1 turnover 24:31 turnover 24:31 turnover of cyclooxygenase-2 24:51 Glycosylation regulates turnover of cyclooxygenase-2. 17113084 3 17 theme biological 382:391 arg1 significance 393:404 the biological significance 378:404 the biological significance of this variable glycosylation 378:435 In this study, Asn(580) was mutated to determine the biological significance of this variable glycosylation. 17113084 1 18 theme prostanoid 119:128 arg1 pathway 143:149 the prostanoid biosynthesis pathway 115:149 the prostanoid biosynthesis pathway 115:149 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 17113084 1 19 theme biosynthesis 130:141 arg1 pathway 143:149 the prostanoid biosynthesis pathway 115:149 the prostanoid biosynthesis pathway 115:149 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 17113084 4 20 theme COX-2 560:564 arg1 protein 566:572 more COX-2 protein 555:572 more COX-2 protein 555:572 COS-1 cells transfected with the mutant gene were unable to express the 74kDa glycoform and were found to accumulate more COX-2 protein and have five times greater COX-2 activity than cells expressing both glycoforms. 17113084 1 21 from step 107:110 arg1 pathway 143:149 the prostanoid biosynthesis pathway 115:149 the prostanoid biosynthesis pathway 115:149 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 17113084 1 22 theme arachidonic 163:173 arg1 acid 175:178 arachidonic acid 163:178 arachidonic acid 163:178 Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in the prostanoid biosynthesis pathway, converting arachidonic acid into prostaglandin H(2). 9153399 4 0 theme domain-containing 605:621 arg1 integrin 623:630 an I domain-containing integrin 600:630 an I domain-containing integrin 600:630 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 0 theme domain-containing 605:621 arg1 antigen 664:670 lymphocyte-function-associated antigen 1 633:672 lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18) 633:695 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 1 1 from Recognition 80:90 arg1 surface 125:131 the cell surface 116:131 the cell surface 116:131 Recognition by integrin proteins on the cell surface regulates the adhesive interactions between cells and their surroundings. 9153399 6 2 theme I 896:896 arg1 domain 898:903 the I domain 892:903 the I domain 892:903 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 8 3 from arrangement 1293:1303 arg1 region 1350:1355 the membrane-proximal region 1328:1355 the membrane-proximal region of domain 2 1328:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 4 4 theme adhesion 470:477 arg1 molecules 479:487 The intercellular adhesion molecules 452:487 The intercellular adhesion molecules ICAM-1, 2 and 3 452:503 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 4 theme adhesion 470:477 arg1 members 523:529 members 523:529 members of the immunoglobulin superfamily (IgSF) 523:570 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 5 theme I 603:603 arg1 integrin 623:630 an I domain-containing integrin 600:630 an I domain-containing integrin 600:630 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 5 theme I 603:603 arg1 antigen 664:670 lymphocyte-function-associated antigen 1 633:672 lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18) 633:695 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 3 6 theme only 318:321 arg1 VCAM-1 395:400 VCAM-1 395:400 VCAM-1 395:400 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 3 6 theme only 318:321 arg1 ligands 332:338 the only integrin ligands 314:338 the only integrin ligands for which structures are known 314:369 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 3 6 theme only 318:321 arg1 fibronectin 379:389 fibronectin 379:389 fibronectin 379:389 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 4 7 theme intercellular 456:468 arg1 molecules 479:487 The intercellular adhesion molecules 452:487 The intercellular adhesion molecules ICAM-1, 2 and 3 452:503 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 7 theme intercellular 456:468 arg1 members 523:529 members 523:529 members of the immunoglobulin superfamily (IgSF) 523:570 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 3 8 theme integrin 323:330 arg1 VCAM-1 395:400 VCAM-1 395:400 VCAM-1 395:400 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 3 8 theme integrin 323:330 arg1 ligands 332:338 the only integrin ligands 314:338 the only integrin ligands for which structures are known 314:369 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 3 8 theme integrin 323:330 arg1 fibronectin 379:389 fibronectin 379:389 fibronectin 379:389 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 6 9 from residue 1035:1041 arg1 fibronectin 1057:1067 fibronectin 1057:1067 fibronectin 1057:1067 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 9 from residue 1035:1041 arg1 VCAM-1 1046:1051 VCAM-1 1046:1051 VCAM-1 1046:1051 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 10 theme recognition 953:963 arg1 surface 965:971 a relatively flat recognition surface 935:971 a relatively flat recognition surface 935:971 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 11 from position 804:811 arg1 critical 819:826 critical 819:826 critical 819:826 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 11 from position 804:811 arg1 residue 793:799 The glutamic acid residue 775:799 The glutamic acid residue at position 37 775:814 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 12 from ion 885:887 arg1 domain 898:903 the I domain 892:903 the I domain 892:903 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 7 13 from differences 1131:1141 arg1 architecture 1150:1161 the architecture 1146:1161 the architecture of recognition sites 1146:1182 This finding suggests that there are differences in the architecture of recognition sites between integrins that contain or lack I domains. 9153399 6 14 theme flat 948:951 arg1 surface 965:971 a relatively flat recognition surface 935:971 a relatively flat recognition surface 935:971 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 5 15 theme crystal 718:724 arg1 structure 726:734 the crystal structure 714:734 the crystal structure of the extracellular region of ICAM-2 714:772 Here we present the crystal structure of the extracellular region of ICAM-2. 9153399 6 16 theme acid 1030:1033 arg1 residue 1035:1041 the critical aspartic acid residue 1008:1041 the critical aspartic acid residue in VCAM-1 and fibronectin 1008:1067 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 8 17 theme recognition 1405:1415 arg1 surface 1417:1423 the recognition surface 1401:1423 the recognition surface to LFA-1 1401:1432 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 4 18 dep antigen 664:670 arg1 LFA-1 675:679 LFA-1 675:679 LFA-1 675:679 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 18 dep antigen 664:670 arg1 CD11a/CD18 685:694 CD11a/CD18 685:694 CD11a/CD18 685:694 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 0 19 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of ICAM-2 0:26 Crystal structure of ICAM-2 reveals a distinctive integrin recognition surface. 9153399 6 20 theme Mg2+ 880:883 arg1 ion 885:887 the Mg2+ ion 876:887 the Mg2+ ion in the I domain 876:903 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 8 21 theme membrane-proximal 1332:1348 arg1 region 1350:1355 the membrane-proximal region 1328:1355 the membrane-proximal region of domain 2 1328:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 4 22 dep molecules 479:487 arg1 3 503:503 3 503:503 3 503:503 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 22 dep molecules 479:487 arg1 2 497:497 2 497:497 2 497:497 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 22 dep molecules 479:487 arg1 ICAM-1 489:494 ICAM-1 489:494 ICAM-1 489:494 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 1 23 theme integrin 95:102 arg1 proteins 104:111 integrin proteins 95:111 integrin proteins 95:111 Recognition by integrin proteins on the cell surface regulates the adhesive interactions between cells and their surroundings. 9153399 0 24 theme ICAM-2 21:26 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of ICAM-2 0:26 Crystal structure of ICAM-2 reveals a distinctive integrin recognition surface. 9153399 6 25 theme critical 1012:1019 arg1 residue 1035:1041 the critical aspartic acid residue 1008:1041 the critical aspartic acid residue in VCAM-1 and fibronectin 1008:1067 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 5 26 theme extracellular 743:755 arg1 region 757:762 the extracellular region 739:762 the extracellular region of ICAM-2 739:772 Here we present the crystal structure of the extracellular region of ICAM-2. 9153399 6 27 theme aspartic 1021:1028 arg1 residue 1035:1041 the critical aspartic acid residue 1008:1041 the critical aspartic acid residue in VCAM-1 and fibronectin 1008:1067 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 28 theme acid 788:791 arg1 critical 819:826 critical 819:826 critical 819:826 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 28 theme acid 788:791 arg1 residue 793:799 The glutamic acid residue 775:799 The glutamic acid residue at position 37 775:814 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 5 29 theme region 757:762 arg1 structure 726:734 the crystal structure 714:734 the crystal structure of the extracellular region of ICAM-2 714:772 Here we present the crystal structure of the extracellular region of ICAM-2. 9153399 7 30 theme recognition 1166:1176 arg1 sites 1178:1182 recognition sites 1166:1182 recognition sites 1166:1182 This finding suggests that there are differences in the architecture of recognition sites between integrins that contain or lack I domains. 9153399 6 31 theme glutamic 779:786 arg1 critical 819:826 critical 819:826 critical 819:826 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 6 31 theme glutamic 779:786 arg1 residue 793:799 The glutamic acid residue 775:799 The glutamic acid residue at position 37 775:814 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 9 32 theme ICAM-2 1466:1471 arg1 structure 1473:1481 the ICAM-2 structure 1462:1481 the ICAM-2 structure 1462:1481 A model of ICAM-1 based on the ICAM-2 structure provides a framework for understanding its recognition by pathogens. 9153399 8 33 theme glycans 1317:1323 arg1 arrangement 1293:1303 a tripod-like arrangement 1279:1303 a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 1279:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 8 33 theme glycans 1317:1323 arg1 glycans 1317:1323 N-linked glycans 1308:1323 N-linked glycans 1308:1323 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 8 33 theme glycans 1317:1323 arg1 ICAM-2 1268:1273 ICAM-2 1268:1273 ICAM-2 1268:1273 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 8 33 theme glycans 1317:1323 arg1 domains 1249:1255 domains 1 and 2 1249:1263 domains 1249:1255 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 2 34 theme I 229:229 arg1 domain 232:237 the 'I' domain 224:237 the 'I' domain that is found in some but not all integrins 224:281 The structure of the 'I' domain that is found in some but not all integrins, has been determined. 9153399 5 35 theme ICAM-2 767:772 arg1 region 757:762 the extracellular region 739:762 the extracellular region of ICAM-2 739:772 Here we present the crystal structure of the extracellular region of ICAM-2. 9153399 0 36 theme integrin 50:57 arg1 surface 71:77 a distinctive integrin recognition surface 36:77 a distinctive integrin recognition surface 36:77 Crystal structure of ICAM-2 reveals a distinctive integrin recognition surface. 9153399 8 37 theme N-linked 1308:1315 arg1 glycans 1317:1323 N-linked glycans 1308:1323 N-linked glycans 1308:1323 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 3 38 theme I 441:441 arg1 domains 443:449 I domains 441:449 I domains 441:449 However, the only integrin ligands for which structures are known, namely fibronectin and VCAM-1, are recognized by integrins that lack I domains. 9153399 0 39 theme distinctive 38:48 arg1 surface 71:77 a distinctive integrin recognition surface 36:77 a distinctive integrin recognition surface 36:77 Crystal structure of ICAM-2 reveals a distinctive integrin recognition surface. 9153399 8 40 theme domain 1360:1365 arg1 region 1350:1355 the membrane-proximal region 1328:1355 the membrane-proximal region of domain 2 1328:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 1 41 theme cell 120:123 arg1 surface 125:131 the cell surface 116:131 the cell surface 116:131 Recognition by integrin proteins on the cell surface regulates the adhesive interactions between cells and their surroundings. 9153399 7 42 theme I 1223:1223 arg1 domains 1225:1231 I domains 1223:1231 I domains 1223:1231 This finding suggests that there are differences in the architecture of recognition sites between integrins that contain or lack I domains. 9153399 6 43 theme protruding 1076:1085 arg1 loops 1087:1091 protruding loops 1076:1091 protruding loops 1076:1091 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 9 44 theme ICAM-1 1446:1451 arg1 model 1437:1441 A model 1435:1441 A model of ICAM-1 based on the ICAM-2 structure 1435:1481 A model of ICAM-1 based on the ICAM-2 structure provides a framework for understanding its recognition by pathogens. 9153399 8 45 theme ICAM-2 1268:1273 arg1 arrangement 1293:1303 a tripod-like arrangement 1279:1303 a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 1279:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 8 45 theme ICAM-2 1268:1273 arg1 glycans 1317:1323 N-linked glycans 1308:1323 N-linked glycans 1308:1323 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 8 45 theme ICAM-2 1268:1273 arg1 ICAM-2 1268:1273 ICAM-2 1268:1273 ICAM-2 1268:1273 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 8 45 theme ICAM-2 1268:1273 arg1 domains 1249:1255 domains 1 and 2 1249:1263 domains 1249:1255 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 4 46 theme lymphocyte-function-associated 633:662 arg1 integrin 623:630 an I domain-containing integrin 600:630 an I domain-containing integrin 600:630 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 46 theme lymphocyte-function-associated 633:662 arg1 antigen 664:670 lymphocyte-function-associated antigen 1 633:672 lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18) 633:695 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 2 47 located found 247:251 arg1 integrins 273:281 all integrins 269:281 some but not all integrins 256:281 The structure of the 'I' domain that is found in some but not all integrins, has been determined. 9153399 2 47 located found 247:251 arg2 domain 232:237 the 'I' domain 224:237 the 'I' domain that is found in some but not all integrins 224:281 The structure of the 'I' domain that is found in some but not all integrins, has been determined. 9153399 2 47 located found 247:251 arg1 some 256:259 some 256:259 some 256:259 The structure of the 'I' domain that is found in some but not all integrins, has been determined. 9153399 7 48 contain contain 1207:1213 arg1 integrins 1192:1200 integrins 1192:1200 integrins that contain or lack I domains 1192:1231 This finding suggests that there are differences in the architecture of recognition sites between integrins that contain or lack I domains. 9153399 7 48 contain contain 1207:1213 arg2 domains 1225:1231 I domains 1223:1231 I domains 1223:1231 This finding suggests that there are differences in the architecture of recognition sites between integrins that contain or lack I domains. 9153399 8 49 theme tripod-like 1281:1291 arg1 arrangement 1293:1303 a tripod-like arrangement 1279:1303 a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 1279:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 7 50 theme sites 1178:1182 arg1 architecture 1150:1161 the architecture 1146:1161 the architecture of recognition sites 1146:1182 This finding suggests that there are differences in the architecture of recognition sites between integrins that contain or lack I domains. 9153399 8 51 link N-linked 1308:1315 arg1 glycans 1317:1323 N-linked glycans 1308:1323 N-linked glycans 1308:1323 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 4 52 theme superfamily 553:563 arg1 molecules 479:487 The intercellular adhesion molecules 452:487 The intercellular adhesion molecules ICAM-1, 2 and 3 452:503 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 52 theme superfamily 553:563 arg1 members 523:529 members 523:529 members of the immunoglobulin superfamily (IgSF) 523:570 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 0 53 theme recognition 59:69 arg1 surface 71:77 a distinctive integrin recognition surface 36:77 a distinctive integrin recognition surface 36:77 Crystal structure of ICAM-2 reveals a distinctive integrin recognition surface. 9153399 6 54 theme LFA-1 832:836 arg1 binding 838:844 LFA-1 binding 832:844 LFA-1 binding 832:844 The glutamic acid residue at position 37 is critical for LFA-1 binding and is proposed to coordinate the Mg2+ ion in the I domain; this Glu 37 is surrounded by a relatively flat recognition surface and lies in a beta-strand, whereas the critical aspartic acid residue in VCAM-1 and fibronectin lie in protruding loops. 9153399 2 55 theme domain 232:237 arg1 structure 211:219 The structure 207:219 The structure 207:219 The structure of the 'I' domain that is found in some but not all integrins, has been determined. 9153399 1 56 theme adhesive 147:154 arg1 interactions 156:167 the adhesive interactions 143:167 the adhesive interactions between cells and their surroundings 143:204 Recognition by integrin proteins on the cell surface regulates the adhesive interactions between cells and their surroundings. 9153399 8 57 from domains 1249:1255 arg1 region 1350:1355 the membrane-proximal region 1328:1355 the membrane-proximal region of domain 2 1328:1367 A bend between domains 1 and 2 of ICAM-2 and a tripod-like arrangement of N-linked glycans in the membrane-proximal region of domain 2 may be important for presenting the recognition surface to LFA-1. 9153399 4 58 theme immunoglobulin 538:551 arg1 IgSF 566:569 IgSF 566:569 IgSF 566:569 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 9153399 4 58 theme immunoglobulin 538:551 arg1 superfamily 553:563 the immunoglobulin superfamily 534:563 the immunoglobulin superfamily (IgSF) 534:570 The intercellular adhesion molecules ICAM-1, 2 and 3 are, like VCAM-1, members of the immunoglobulin superfamily (IgSF), but they are recognized by an I domain-containing integrin, lymphocyte-function-associated antigen 1 (LFA-1, or CD11a/CD18). 23846701 8 0 theme binding 911:917 arg1 motif 919:923 A central binding motif 901:923 A central binding motif 901:923 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 8 1 theme corrin 981:986 arg1 ring 988:991 the corrin ring 977:991 the corrin ring 977:991 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 9 2 theme several 1103:1109 arg1 contacts 1130:1137 several unique interdomain contacts 1103:1137 several unique interdomain contacts 1103:1137 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 1 3 theme essential 136:144 arg1 Cobalamin 101:109 Cobalamin 101:109 Cobalamin (Cbl; vitamin B12) 101:128 Cobalamin (Cbl; vitamin B12) is an essential micronutrient synthesized only by bacteria. 23846701 1 3 theme essential 136:144 arg1 micronutrient 146:158 an essential micronutrient 133:158 an essential micronutrient synthesized only by bacteria 133:187 Cobalamin (Cbl; vitamin B12) is an essential micronutrient synthesized only by bacteria. 23846701 11 4 theme Cbl 1485:1487 arg1 interaction 1489:1499 Cbl interaction 1485:1499 Cbl interaction with its three transport proteins 1485:1533 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 0 5 theme protein 80:86 arg1 haptocorrin 88:98 the cobalamin transport protein haptocorrin 56:98 the cobalamin transport protein haptocorrin 56:98 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin. 23846701 9 6 theme higher 1150:1155 arg1 complementarity 1163:1177 a higher shape complementarity 1148:1177 a higher shape complementarity than those of intrinsic factor and transcobalamin 1148:1227 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 9 7 theme unique 1111:1116 arg1 contacts 1130:1137 several unique interdomain contacts 1103:1137 several unique interdomain contacts 1103:1137 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 1 8 dep Cobalamin 101:109 arg1 Cbl 112:114 Cbl 112:114 Cbl; vitamin B12 112:127 Cobalamin (Cbl; vitamin B12) is an essential micronutrient synthesized only by bacteria. 23846701 7 9 theme distinct 822:829 arg1 interactions 831:842 distinct interactions 822:842 distinct interactions allowing for stabilization of corrinoids other than Cbl 822:898 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 11 10 theme structural 1369:1378 arg1 analysis 1380:1387 Our structural analysis 1365:1387 Our structural analysis 1365:1387 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 9 11 theme interdomain 1118:1128 arg1 contacts 1130:1137 several unique interdomain contacts 1103:1137 several unique interdomain contacts 1103:1137 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 3 12 theme intrinsic 352:360 arg1 factor 362:367 intrinsic factor 352:367 intrinsic factor 352:367 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 12 theme intrinsic 352:360 arg1 proteins 326:333 three transport proteins 310:333 three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC) 310:389 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 8 13 theme ring 988:991 arg1 chains 967:972 the e- and f-side chains 949:972 the e- and f-side chains of the corrin ring 949:991 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 8 14 theme e- 953:954 arg1 chains 967:972 the e- and f-side chains 949:972 the e- and f-side chains of the corrin ring 949:991 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 6 15 theme interactions 669:680 arg1 many 657:660 many 657:660 many 657:660 The structures reveal that many of the interactions with the corrin ring are conserved among the human Cbl transporters. 23846701 6 15 theme interactions 669:680 arg1 interactions 669:680 the interactions 665:680 the interactions with the corrin ring 665:701 The structures reveal that many of the interactions with the corrin ring are conserved among the human Cbl transporters. 23846701 9 16 contain have 1143:1146 arg2 complementarity 1163:1177 a higher shape complementarity 1148:1177 a higher shape complementarity than those of intrinsic factor and transcobalamin 1148:1227 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 9 16 contain have 1143:1146 arg1 α- 1075:1076 α- 1075:1076 α- 1075:1076 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 5 17 attach present 497:503 arg1 resolution 604:613 2.35 and 3.0 Å resolution 589:613 2.35 and 3.0 Å resolution 589:613 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 5 17 attach present 497:503 arg2 we 494:495 we 494:495 we 494:495 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 8 18 with interactions 931:942 arg1 chains 967:972 the e- and f-side chains 949:972 the e- and f-side chains of the corrin ring 949:991 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 5 19 theme HC 537:538 arg1 structures 517:526 the crystal structures 505:526 the crystal structures of human HC in complex with cyanocobalamin and cobinamide 505:584 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 8 20 theme species 1049:1055 arg1 proteins 1031:1038 corrinoid-binding proteins 1013:1038 corrinoid-binding proteins of other species 1013:1055 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 4 21 contain have 411:414 arg1 proteins 402:409 All three proteins 392:409 All three proteins 392:409 All three proteins have a similar overall structure but a different selectivity for corrinoids. 23846701 4 21 contain have 411:414 arg2 structure 434:442 a similar overall structure 416:442 a similar overall structure 416:442 All three proteins have a similar overall structure but a different selectivity for corrinoids. 23846701 4 21 contain have 411:414 arg2 selectivity 460:470 a different selectivity 448:470 a different selectivity for corrinoids 448:485 All three proteins have a similar overall structure but a different selectivity for corrinoids. 23846701 4 22 theme different 450:458 arg1 selectivity 460:470 a different selectivity 448:470 a different selectivity for corrinoids 448:485 All three proteins have a similar overall structure but a different selectivity for corrinoids. 23846701 8 23 theme corrinoid-binding 1013:1029 arg1 proteins 1031:1038 corrinoid-binding proteins 1013:1038 corrinoid-binding proteins of other species 1013:1055 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 0 24 theme transport 70:78 arg1 haptocorrin 88:98 the cobalamin transport protein haptocorrin 56:98 the cobalamin transport protein haptocorrin 56:98 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin. 23846701 9 25 dep α- 1075:1076 arg1 the 1071:1073 the 1071:1073 the 1071:1073 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 0 26 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin 0:98 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin. 23846701 5 27 with complex 543:549 arg1 cobinamide 575:584 cobinamide 575:584 cobinamide 575:584 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 5 27 with complex 543:549 arg1 cyanocobalamin 556:569 cyanocobalamin 556:569 cyanocobalamin 556:569 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 5 28 theme Å 602:602 arg1 resolution 604:613 2.35 and 3.0 Å resolution 589:613 2.35 and 3.0 Å resolution 589:613 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 6 29 theme Cbl 733:735 arg1 transporters 737:748 the human Cbl transporters 723:748 the human Cbl transporters 723:748 The structures reveal that many of the interactions with the corrin ring are conserved among the human Cbl transporters. 23846701 11 30 theme transport 1516:1524 arg1 proteins 1526:1533 three transport proteins 1510:1533 its three transport proteins 1506:1533 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 11 31 theme binding 1433:1439 arg1 behavior 1441:1448 the unique binding behavior 1422:1448 the unique binding behavior of HC 1422:1454 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 0 32 theme universal 21:29 arg1 recognition 41:51 universal corrinoid recognition 21:51 universal corrinoid recognition by the cobalamin transport protein haptocorrin 21:98 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin. 23846701 7 33 theme non-conserved 764:776 arg1 residues 778:785 the non-conserved residues 760:785 the non-conserved residues Asn-120, Arg-357, and Asn-373 760:815 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 7 33 theme non-conserved 764:776 arg1 Arg-357 796:802 Arg-357 796:802 Arg-357 796:802 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 7 33 theme non-conserved 764:776 arg1 Asn-373 809:815 Asn-373 809:815 Asn-373 809:815 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 7 33 theme non-conserved 764:776 arg1 Asn-120 787:793 Asn-120 787:793 Asn-120 787:793 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 6 34 theme human 727:731 arg1 transporters 737:748 the human Cbl transporters 723:748 the human Cbl transporters 723:748 The structures reveal that many of the interactions with the corrin ring are conserved among the human Cbl transporters. 23846701 10 35 theme complexes 1354:1362 arg1 temperatures 1319:1330 higher melting temperatures 1304:1330 higher melting temperatures of the protein-ligand complexes 1304:1362 The stabilization of ligands by all of these interactions is reflected in higher melting temperatures of the protein-ligand complexes. 23846701 8 36 theme central 903:909 arg1 motif 919:923 A central binding motif 901:923 A central binding motif 901:923 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 4 37 theme similar 418:424 arg1 structure 434:442 a similar overall structure 416:442 a similar overall structure 416:442 All three proteins have a similar overall structure but a different selectivity for corrinoids. 23846701 11 38 theme unique 1426:1431 arg1 behavior 1441:1448 the unique binding behavior 1422:1448 the unique binding behavior of HC 1422:1454 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 6 39 with interactions 669:680 arg1 ring 698:701 the corrin ring 687:701 the corrin ring 687:701 The structures reveal that many of the interactions with the corrin ring are conserved among the human Cbl transporters. 23846701 10 40 theme ligands 1251:1257 arg1 stabilization 1234:1246 The stabilization 1230:1246 The stabilization of ligands by all of these interactions 1230:1286 The stabilization of ligands by all of these interactions is reflected in higher melting temperatures of the protein-ligand complexes. 23846701 0 41 theme corrinoid 31:39 arg1 recognition 41:51 universal corrinoid recognition 21:51 universal corrinoid recognition by the cobalamin transport protein haptocorrin 21:98 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin. 23846701 11 42 with interaction 1489:1499 arg1 proteins 1526:1533 three transport proteins 1510:1533 its three transport proteins 1506:1533 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 3 43 dep proteins 326:333 arg1 transcobalamin 336:349 transcobalamin 336:349 transcobalamin 336:349 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 43 dep proteins 326:333 arg1 factor 362:367 intrinsic factor 352:367 intrinsic factor 352:367 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 43 dep proteins 326:333 arg1 haptocorrin 374:384 haptocorrin 374:384 haptocorrin (HC) 374:389 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 43 dep proteins 326:333 arg1 proteins 326:333 three transport proteins 310:333 three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC) 310:389 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 43 dep proteins 326:333 arg1 HC 387:388 HC 387:388 HC 387:388 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 2 44 theme uptake 229:234 arg1 system 236:241 a sophisticated uptake system 213:241 a sophisticated uptake system to capture the vitamin from the diet 213:278 Mammals have developed a sophisticated uptake system to capture the vitamin from the diet. 23846701 5 45 theme crystal 509:515 arg1 structures 517:526 the crystal structures 505:526 the crystal structures of human HC in complex with cyanocobalamin and cobinamide 505:584 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 5 46 from structures 517:526 arg1 complex 543:549 complex 543:549 complex with cyanocobalamin and cobinamide 543:584 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 4 47 theme overall 426:432 arg1 structure 434:442 a similar overall structure 416:442 a similar overall structure 416:442 All three proteins have a similar overall structure but a different selectivity for corrinoids. 23846701 2 48 theme sophisticated 215:227 arg1 system 236:241 a sophisticated uptake system 213:241 a sophisticated uptake system to capture the vitamin from the diet 213:278 Mammals have developed a sophisticated uptake system to capture the vitamin from the diet. 23846701 8 49 theme other 1043:1047 arg1 species 1049:1055 other species 1043:1055 other species 1043:1055 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 10 50 theme protein-ligand 1339:1352 arg1 complexes 1354:1362 the protein-ligand complexes 1335:1362 the protein-ligand complexes 1335:1362 The stabilization of ligands by all of these interactions is reflected in higher melting temperatures of the protein-ligand complexes. 23846701 11 51 theme fundamental 1396:1406 arg1 insights 1408:1415 fundamental insights 1396:1415 fundamental insights into the unique binding behavior of HC 1396:1454 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 9 52 theme intrinsic 1193:1201 arg1 factor 1203:1208 intrinsic factor 1193:1208 intrinsic factor 1193:1208 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 11 53 theme HC 1453:1454 arg1 behavior 1441:1448 the unique binding behavior 1422:1448 the unique binding behavior of HC 1422:1454 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 1 54 theme vitamin 117:123 arg1 B12 125:127 vitamin B12 117:127 Cbl; vitamin B12 112:127 Cobalamin (Cbl; vitamin B12) is an essential micronutrient synthesized only by bacteria. 23846701 7 55 theme corrinoids 874:883 arg1 stabilization 857:869 stabilization 857:869 stabilization of corrinoids other than Cbl 857:898 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 9 56 theme shape 1157:1161 arg1 complementarity 1163:1177 a higher shape complementarity 1148:1177 a higher shape complementarity than those of intrinsic factor and transcobalamin 1148:1227 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 3 57 theme Cbl 281:283 arg1 transport 285:293 Cbl transport 281:293 Cbl transport 281:293 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 8 58 theme f-side 960:965 arg1 chains 967:972 the e- and f-side chains 949:972 the e- and f-side chains of the corrin ring 949:991 A central binding motif forms interactions with the e- and f-side chains of the corrin ring and is conserved in corrinoid-binding proteins of other species. 23846701 11 59 theme interaction 1489:1499 arg1 picture 1474:1480 the picture 1470:1480 the picture of Cbl interaction with its three transport proteins 1470:1533 Our structural analysis offers fundamental insights into the unique binding behavior of HC and completes the picture of Cbl interaction with its three transport proteins. 23846701 5 60 theme human 531:535 arg1 HC 537:538 human HC 531:538 human HC 531:538 Here, we present the crystal structures of human HC in complex with cyanocobalamin and cobinamide at 2.35 and 3.0 Å resolution, respectively. 23846701 7 61 theme other 885:889 arg1 corrinoids 874:883 corrinoids 874:883 corrinoids other than Cbl 874:898 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 9 62 theme HC 1095:1096 arg1 β-domains 1082:1090 β-domains 1082:1090 β-domains 1082:1090 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 9 62 theme HC 1095:1096 arg1 α- 1075:1076 α- 1075:1076 α- 1075:1076 In addition, the α- and β-domains of HC form several unique interdomain contacts and have a higher shape complementarity than those of intrinsic factor and transcobalamin. 23846701 7 63 dep residues 778:785 arg1 residues 778:785 the non-conserved residues 760:785 the non-conserved residues Asn-120, Arg-357, and Asn-373 760:815 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 7 63 dep residues 778:785 arg1 Arg-357 796:802 Arg-357 796:802 Arg-357 796:802 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 7 63 dep residues 778:785 arg1 Asn-373 809:815 Asn-373 809:815 Asn-373 809:815 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 7 63 dep residues 778:785 arg1 Asn-120 787:793 Asn-120 787:793 Asn-120 787:793 However, the non-conserved residues Asn-120, Arg-357, and Asn-373 form distinct interactions allowing for stabilization of corrinoids other than Cbl. 23846701 3 64 theme transport 316:324 arg1 transcobalamin 336:349 transcobalamin 336:349 transcobalamin 336:349 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 64 theme transport 316:324 arg1 factor 362:367 intrinsic factor 352:367 intrinsic factor 352:367 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 64 theme transport 316:324 arg1 proteins 326:333 three transport proteins 310:333 three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC) 310:389 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 3 64 theme transport 316:324 arg1 haptocorrin 374:384 haptocorrin 374:384 haptocorrin (HC) 374:389 Cbl transport is mediated by three transport proteins: transcobalamin, intrinsic factor, and haptocorrin (HC). 23846701 10 65 theme melting 1311:1317 arg1 temperatures 1319:1330 higher melting temperatures 1304:1330 higher melting temperatures of the protein-ligand complexes 1304:1362 The stabilization of ligands by all of these interactions is reflected in higher melting temperatures of the protein-ligand complexes. 23846701 6 66 theme corrin 691:696 arg1 ring 698:701 the corrin ring 687:701 the corrin ring 687:701 The structures reveal that many of the interactions with the corrin ring are conserved among the human Cbl transporters. 23846701 2 67 from diet 275:278 arg1 vitamin 258:264 the vitamin 254:264 the vitamin from the diet 254:278 Mammals have developed a sophisticated uptake system to capture the vitamin from the diet. 23846701 0 68 theme cobalamin 60:68 arg1 haptocorrin 88:98 the cobalamin transport protein haptocorrin 56:98 the cobalamin transport protein haptocorrin 56:98 Structural basis for universal corrinoid recognition by the cobalamin transport protein haptocorrin. 23846701 10 69 theme higher 1304:1309 arg1 temperatures 1319:1330 higher melting temperatures 1304:1330 higher melting temperatures of the protein-ligand complexes 1304:1362 The stabilization of ligands by all of these interactions is reflected in higher melting temperatures of the protein-ligand complexes. 23846701 1 70 dep Cbl 112:114 arg1 B12 125:127 vitamin B12 117:127 Cbl; vitamin B12 112:127 Cobalamin (Cbl; vitamin B12) is an essential micronutrient synthesized only by bacteria. 12620237 5 0 theme dimerization 817:828 arg1 interface 830:838 this dimerization interface 812:838 this dimerization interface 812:838 To activate the receptor, EGF binding must promote a large domain rearrangement that exposes this dimerization interface. 12620237 6 1 theme new 914:916 arg1 approaches 918:927 new approaches 914:927 new approaches for designing ErbB receptor antagonists 914:967 This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists. 12620237 2 2 theme EGF 372:374 arg1 region 399:404 the 621 amino acid EGF receptor extracellular region 353:404 the 621 amino acid EGF receptor extracellular region 353:404 EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. 12620237 3 3 theme entire 466:471 arg1 sEGFR 495:499 sEGFR 495:499 sEGFR 495:499 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 3 3 theme entire 466:471 arg1 region 487:492 this entire extracellular region 461:492 this entire extracellular region (sEGFR) 461:500 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 1 4 theme cell 224:227 arg1 growth 229:234 cell growth 224:234 cell growth 224:234 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 3 5 theme A 427:427 arg1 structure 448:456 the 2.8 A resolution crystal structure 419:456 the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state 419:524 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 1 6 theme ErbB 157:160 arg1 RTKs 202:205 RTKs 202:205 RTKs 202:205 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 6 theme ErbB 157:160 arg1 kinases 193:199 the ErbB (HER) family receptor tyrosine kinases 153:199 the ErbB (HER) family receptor tyrosine kinases (RTKs) 153:206 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 6 7 theme activation 879:888 arg1 mechanisms 890:899 other RTK activation mechanisms 869:899 other RTK activation mechanisms 869:899 This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists. 12620237 6 8 theme ErbB 943:946 arg1 antagonists 957:967 ErbB receptor antagonists 943:967 ErbB receptor antagonists 943:967 This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists. 12620237 6 9 theme RTK 875:877 arg1 mechanisms 890:899 other RTK activation mechanisms 869:899 other RTK activation mechanisms 869:899 This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists. 12620237 1 10 theme Epidermal 94:102 arg1 EGF 119:121 EGF 119:121 EGF 119:121 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 10 theme Epidermal 94:102 arg1 factor 111:116 Epidermal growth factor 94:116 Epidermal growth factor (EGF) receptor 94:131 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 4 11 theme sEGFR 647:651 arg1 structures 653:662 activated sEGFR structures 637:662 activated sEGFR structures 637:662 The structure reveals an autoinhibited configuration, where the dimerization interface recently identified in activated sEGFR structures is completely occluded by intramolecular interactions. 12620237 3 12 from structure 448:456 arg1 state 520:524 an unactivated state 505:524 an unactivated state 505:524 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 1 13 theme growth 104:109 arg1 EGF 119:121 EGF 119:121 EGF 119:121 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 13 theme growth 104:109 arg1 factor 111:116 Epidermal growth factor 94:116 Epidermal growth factor (EGF) receptor 94:131 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 4 14 theme activated 637:645 arg1 structures 653:662 activated sEGFR structures 637:662 activated sEGFR structures 637:662 The structure reveals an autoinhibited configuration, where the dimerization interface recently identified in activated sEGFR structures is completely occluded by intramolecular interactions. 12620237 2 15 theme region 399:404 arg1 dimerization 337:348 dimerization 337:348 dimerization of the 621 amino acid EGF receptor extracellular region 337:404 EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. 12620237 1 16 theme factor 111:116 arg1 receptor 124:131 Epidermal growth factor (EGF) receptor 94:131 Epidermal growth factor (EGF) receptor 94:131 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 16 theme factor 111:116 arg1 prototype 140:148 the prototype 136:148 the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers 136:295 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 2 17 theme receptor 376:383 arg1 region 399:404 the 621 amino acid EGF receptor extracellular region 353:404 the 621 amino acid EGF receptor extracellular region 353:404 EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. 12620237 2 18 theme extracellular 385:397 arg1 region 399:404 the 621 amino acid EGF receptor extracellular region 353:404 the 621 amino acid EGF receptor extracellular region 353:404 EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. 12620237 6 19 theme other 869:873 arg1 mechanisms 890:899 other RTK activation mechanisms 869:899 other RTK activation mechanisms 869:899 This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists. 12620237 3 20 theme unactivated 508:518 arg1 state 520:524 an unactivated state 505:524 an unactivated state 505:524 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 1 21 theme receptor 175:182 arg1 RTKs 202:205 RTKs 202:205 RTKs 202:205 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 21 theme receptor 175:182 arg1 kinases 193:199 the ErbB (HER) family receptor tyrosine kinases 153:199 the ErbB (HER) family receptor tyrosine kinases (RTKs) 153:206 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 22 theme tyrosine 184:191 arg1 RTKs 202:205 RTKs 202:205 RTKs 202:205 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 22 theme tyrosine 184:191 arg1 kinases 193:199 the ErbB (HER) family receptor tyrosine kinases 153:199 the ErbB (HER) family receptor tyrosine kinases (RTKs) 153:206 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 2 23 theme acid 367:370 arg1 region 399:404 the 621 amino acid EGF receptor extracellular region 353:404 the 621 amino acid EGF receptor extracellular region 353:404 EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. 12620237 2 24 theme amino 361:365 arg1 region 399:404 the 621 amino acid EGF receptor extracellular region 353:404 the 621 amino acid EGF receptor extracellular region 353:404 EGF activates its receptor by inducing dimerization of the 621 amino acid EGF receptor extracellular region. 12620237 5 25 theme large 772:776 arg1 rearrangement 785:797 a large domain rearrangement 770:797 a large domain rearrangement that exposes this dimerization interface 770:838 To activate the receptor, EGF binding must promote a large domain rearrangement that exposes this dimerization interface. 12620237 6 26 theme receptor 948:955 arg1 antagonists 957:967 ErbB receptor antagonists 943:967 ErbB receptor antagonists 943:967 This contrasts starkly with other RTK activation mechanisms and suggests new approaches for designing ErbB receptor antagonists. 12620237 1 27 dep ErbB 157:160 arg1 HER 163:165 HER 163:165 HER 163:165 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 4 28 theme intramolecular 690:703 arg1 interactions 705:716 intramolecular interactions 690:716 intramolecular interactions 690:716 The structure reveals an autoinhibited configuration, where the dimerization interface recently identified in activated sEGFR structures is completely occluded by intramolecular interactions. 12620237 5 29 theme domain 778:783 arg1 rearrangement 785:797 a large domain rearrangement 770:797 a large domain rearrangement that exposes this dimerization interface 770:838 To activate the receptor, EGF binding must promote a large domain rearrangement that exposes this dimerization interface. 12620237 1 30 theme many 278:281 arg1 cancers 289:295 many human cancers 278:295 many human cancers 278:295 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 31 theme human 283:287 arg1 cancers 289:295 many human cancers 278:295 many human cancers 278:295 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 3 32 theme resolution 429:438 arg1 structure 448:456 the 2.8 A resolution crystal structure 419:456 the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state 419:524 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 3 33 theme crystal 440:446 arg1 structure 448:456 the 2.8 A resolution crystal structure 419:456 the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state 419:524 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 4 34 theme autoinhibited 552:564 arg1 configuration 566:578 an autoinhibited configuration 549:578 an autoinhibited configuration 549:578 The structure reveals an autoinhibited configuration, where the dimerization interface recently identified in activated sEGFR structures is completely occluded by intramolecular interactions. 12620237 0 35 theme ectodomain 69:78 arg1 dimerization 80:91 ectodomain dimerization 69:91 ectodomain dimerization 69:91 EGF activates its receptor by removing interactions that autoinhibit ectodomain dimerization. 12620237 1 36 theme kinases 193:199 arg1 receptor 124:131 Epidermal growth factor (EGF) receptor 94:131 Epidermal growth factor (EGF) receptor 94:131 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 36 theme kinases 193:199 arg1 prototype 140:148 the prototype 136:148 the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers 136:295 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 37 theme family 168:173 arg1 RTKs 202:205 RTKs 202:205 RTKs 202:205 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 1 37 theme family 168:173 arg1 kinases 193:199 the ErbB (HER) family receptor tyrosine kinases 153:199 the ErbB (HER) family receptor tyrosine kinases (RTKs) 153:206 Epidermal growth factor (EGF) receptor is the prototype of the ErbB (HER) family receptor tyrosine kinases (RTKs), which regulate cell growth and differentiation and are implicated in many human cancers. 12620237 3 38 theme extracellular 473:485 arg1 sEGFR 495:499 sEGFR 495:499 sEGFR 495:499 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 3 38 theme extracellular 473:485 arg1 region 487:492 this entire extracellular region 461:492 this entire extracellular region (sEGFR) 461:500 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 12620237 5 39 theme EGF 745:747 arg1 binding 749:755 EGF binding 745:755 EGF binding 745:755 To activate the receptor, EGF binding must promote a large domain rearrangement that exposes this dimerization interface. 12620237 4 40 theme dimerization 591:602 arg1 interface 604:612 the dimerization interface 587:612 the dimerization interface recently identified in activated sEGFR structures 587:662 The structure reveals an autoinhibited configuration, where the dimerization interface recently identified in activated sEGFR structures is completely occluded by intramolecular interactions. 12620237 3 41 theme region 487:492 arg1 structure 448:456 the 2.8 A resolution crystal structure 419:456 the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state 419:524 We describe the 2.8 A resolution crystal structure of this entire extracellular region (sEGFR) in an unactivated state. 32439656 3 0 from EMC 398:400 arg1 nanodisc 413:420 a lipid nanodisc 405:420 a lipid nanodisc 405:420 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 1 1 theme protein 130:136 arg1 biogenesis 105:114 the biogenesis 101:114 the biogenesis of a membrane protein 101:136 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 0 2 theme protein 65:71 arg1 complex 73:79 the human ER membrane protein complex 43:79 the human ER membrane protein complex 43:79 Structural basis for membrane insertion by the human ER membrane protein complex. 32439656 2 3 theme co- 310:312 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 3 theme co- 310:312 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 3 4 from structure 375:383 arg1 nanodisc 413:420 a lipid nanodisc 405:420 a lipid nanodisc 405:420 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 4 5 theme cytosolic 652:660 arg1 loop 662:665 a methionine-rich cytosolic loop 634:665 a methionine-rich cytosolic loop 634:665 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 2 6 theme membrane 264:271 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 6 theme membrane 264:271 arg1 EMC 290:292 EMC 290:292 EMC 290:292 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 6 theme membrane 264:271 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 4 7 dep subunits 750:757 arg1 EMC6 768:771 EMC6 768:771 EMC6 768:771 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 4 7 dep subunits 750:757 arg1 EMC3 759:762 EMC3 759:762 EMC3 759:762 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 4 7 dep subunits 750:757 arg1 subunits 750:757 the subunits EMC3 and EMC6 746:771 the subunits EMC3 and EMC6 746:771 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 4 8 theme enclosed 685:692 arg1 vestibule 706:714 an enclosed hydrophilic vestibule 682:714 an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6 682:771 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 5 9 theme local 803:807 arg1 thinning 818:825 local membrane thinning 803:825 local membrane thinning 803:825 We propose that the EMC uses local membrane thinning and a positively charged patch to decrease the energetic barrier for insertion into the bilayer. 32439656 3 10 theme overall 428:434 arg1 resolution 436:445 an overall resolution 425:445 an overall resolution of 3.4 angstroms 425:462 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 3 11 theme cryo-electron 467:479 arg1 microscopy 481:490 cryo-electron microscopy 467:490 cryo-electron microscopy 467:490 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 3 12 theme human 392:396 arg1 EMC 398:400 the human EMC 388:400 the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms 388:462 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 3 13 theme EMC 398:400 arg1 structure 375:383 the structure 371:383 the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms 371:462 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 4 14 theme methionine-rich 636:650 arg1 loop 662:665 a methionine-rich cytosolic loop 634:665 a methionine-rich cytosolic loop 634:665 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 0 15 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for membrane insertion by the human ER membrane protein complex. 0:80 Structural basis for membrane insertion by the human ER membrane protein complex. 32439656 0 16 theme membrane 21:28 arg1 insertion 30:38 membrane insertion 21:38 membrane insertion 21:38 Structural basis for membrane insertion by the human ER membrane protein complex. 32439656 1 17 theme defining 84:91 arg1 insertion 145:153 the insertion 141:153 the insertion of its hydrophobic transmembrane helices into the lipid bilayer 141:217 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 1 17 theme defining 84:91 arg1 step 93:96 A defining step 82:96 A defining step in the biogenesis of a membrane protein 82:136 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 1 18 theme hydrophobic 162:172 arg1 helices 188:194 its hydrophobic transmembrane helices 158:194 its hydrophobic transmembrane helices 158:194 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 2 19 theme protein 273:279 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 19 theme protein 273:279 arg1 EMC 290:292 EMC 290:292 EMC 290:292 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 19 theme protein 273:279 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 1 20 theme transmembrane 174:186 arg1 helices 188:194 its hydrophobic transmembrane helices 158:194 its hydrophobic transmembrane helices 158:194 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 4 21 theme substrate 605:613 arg1 insertion 615:623 substrate insertion 605:623 substrate insertion 605:623 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 3 22 theme model 541:545 arg1 building 504:511 building 504:511 building of a nearly complete atomic model 504:545 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 1 23 theme helices 188:194 arg1 insertion 145:153 the insertion 141:153 the insertion of its hydrophobic transmembrane helices into the lipid bilayer 141:217 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 1 23 theme helices 188:194 arg1 step 93:96 A defining step 82:96 A defining step in the biogenesis of a membrane protein 82:136 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 4 24 used used 551:554 arg2 We 548:549 We 548:549 We 548:549 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 5 25 theme membrane 809:816 arg1 thinning 818:825 local membrane thinning 803:825 local membrane thinning 803:825 We propose that the EMC uses local membrane thinning and a positively charged patch to decrease the energetic barrier for insertion into the bilayer. 32439656 0 26 theme human 47:51 arg1 complex 73:79 the human ER membrane protein complex 43:79 the human ER membrane protein complex 43:79 Structural basis for membrane insertion by the human ER membrane protein complex. 32439656 5 27 theme charged 844:850 arg1 patch 852:856 a positively charged patch 831:856 a positively charged patch 831:856 We propose that the EMC uses local membrane thinning and a positively charged patch to decrease the energetic barrier for insertion into the bilayer. 32439656 3 28 theme complete 525:532 arg1 model 541:545 a nearly complete atomic model 516:545 a nearly complete atomic model 516:545 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 2 29 theme conserved 300:308 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 29 theme conserved 300:308 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 3 30 theme atomic 534:539 arg1 model 541:545 a nearly complete atomic model 516:545 a nearly complete atomic model 516:545 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 5 31 theme energetic 874:882 arg1 barrier 884:890 the energetic barrier 870:890 the energetic barrier for insertion into the bilayer 870:921 We propose that the EMC uses local membrane thinning and a positively charged patch to decrease the energetic barrier for insertion into the bilayer. 32439656 2 32 theme reticulum 249:257 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 32 theme reticulum 249:257 arg1 EMC 290:292 EMC 290:292 EMC 290:292 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 32 theme reticulum 249:257 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 33 from ER 353:354 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 33 from ER 353:354 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 4 34 theme hydrophilic 694:704 arg1 vestibule 706:714 an enclosed hydrophilic vestibule 682:714 an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6 682:771 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 32439656 1 35 from step 93:96 arg1 biogenesis 105:114 the biogenesis 101:114 the biogenesis of a membrane protein 101:136 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 3 36 from nanodisc 413:420 arg1 structure 375:383 the structure 371:383 the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms 371:462 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 2 37 theme endoplasmic 237:247 arg1 ER 260:261 ER 260:261 ER 260:261 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 37 theme endoplasmic 237:247 arg1 reticulum 249:257 nine-subunit endoplasmic reticulum 224:257 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 1 38 theme membrane 121:128 arg1 protein 130:136 a membrane protein 119:136 a membrane protein 119:136 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 1 39 theme lipid 205:209 arg1 bilayer 211:217 the lipid bilayer 201:217 the lipid bilayer 201:217 A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. 32439656 0 40 theme membrane 56:63 arg1 complex 73:79 the human ER membrane protein complex 43:79 the human ER membrane protein complex 43:79 Structural basis for membrane insertion by the human ER membrane protein complex. 32439656 2 41 theme nine-subunit 224:235 arg1 ER 260:261 ER 260:261 ER 260:261 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 41 theme nine-subunit 224:235 arg1 reticulum 249:257 nine-subunit endoplasmic reticulum 224:257 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 3 42 theme lipid 407:411 arg1 nanodisc 413:420 a lipid nanodisc 405:420 a lipid nanodisc 405:420 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 3 43 theme angstroms 454:462 arg1 resolution 436:445 an overall resolution 425:445 an overall resolution of 3.4 angstroms 425:462 We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo-electron microscopy, permitting building of a nearly complete atomic model. 32439656 0 44 theme ER 53:54 arg1 complex 73:79 the human ER membrane protein complex 43:79 the human ER membrane protein complex 43:79 Structural basis for membrane insertion by the human ER membrane protein complex. 32439656 2 45 theme posttranslational 318:334 arg1 complex 281:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex 220:287 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) 220:293 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 2 45 theme posttranslational 318:334 arg1 insertase 336:344 a conserved co- and posttranslational insertase 298:344 a conserved co- and posttranslational insertase at the ER 298:354 The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. 32439656 4 46 theme structure-guided 556:571 arg1 mutagenesis 573:583 structure-guided mutagenesis 556:583 structure-guided mutagenesis 556:583 We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. 30765605 6 0 theme subtype-specific 851:866 arg1 blockers 868:875 subtype-specific blockers 851:875 subtype-specific blockers for Nav channels 851:892 The structure establishes a framework for the rational design of subtype-specific blockers for Nav channels. 30765605 2 1 theme μ-conotoxin 333:343 arg1 blocker 320:326 a peptidic pore blocker 304:326 a peptidic pore blocker 304:326 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 1 theme μ-conotoxin 333:343 arg1 KIIIA 345:349 the μ-conotoxin KIIIA 329:349 the μ-conotoxin KIIIA 329:349 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 2 theme angstroms 429:437 arg1 resolution 411:420 an overall resolution 400:420 an overall resolution of 3.0 angstroms 400:437 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 4 3 theme selectivity 665:675 arg1 filter 677:682 the selectivity filter 661:682 the selectivity filter 661:682 The 16-residue KIIIA interacts with the extracellular segments in repeats I to III, placing Lys7 at the entrance to the selectivity filter. 30765605 2 4 theme human 282:286 arg1 Nav1.2 288:293 human Nav1.2 282:293 human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms 282:437 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 3 5 theme pore 507:510 arg1 domain 512:517 the pore domain 503:517 the pore domain 503:517 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 0 6 theme μ-conotoxin 69:79 arg1 KIIIA 81:85 the μ-conotoxin KIIIA 65:85 the μ-conotoxin KIIIA 65:85 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 30765605 6 7 theme rational 832:839 arg1 design 841:846 the rational design 828:846 the rational design of subtype-specific blockers for Nav channels 828:892 The structure establishes a framework for the rational design of subtype-specific blockers for Nav channels. 30765605 5 8 theme KIIIA 767:771 arg1 specificity 773:783 KIIIA specificity 767:783 KIIIA specificity 767:783 Many interacting residues are specific to Nav1.2, revealing a molecular basis for KIIIA specificity. 30765605 2 9 theme subunit 384:390 arg1 presence 359:366 the presence 355:366 the presence of an auxiliary subunit, β2, 355:395 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 10 theme overall 403:409 arg1 resolution 411:420 an overall resolution 400:420 an overall resolution of 3.0 angstroms 400:437 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 11 theme microscopy 258:267 arg1 structure 269:277 the cryo-electron microscopy structure 240:277 the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms 240:437 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 3 12 theme domain 512:517 arg1 shoulder 491:498 the shoulder 487:498 the shoulder of the pore domain 487:517 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 0 13 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 0:86 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 30765605 4 14 from segments 599:606 arg1 repeats 611:617 repeats I to III 611:626 repeats I to III 611:626 The 16-residue KIIIA interacts with the extracellular segments in repeats I to III, placing Lys7 at the entrance to the selectivity filter. 30765605 0 15 theme pore 20:23 arg1 blockade 25:32 pore blockade 20:32 pore blockade of human Na+ channel Nav1.2 20:60 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 30765605 1 16 theme action 181:186 arg1 potentials 188:197 action potentials 181:197 action potentials 181:197 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 2 17 theme Nav1.2 288:293 arg1 structure 269:277 the cryo-electron microscopy structure 240:277 the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms 240:437 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 1 18 theme potentials 188:197 arg1 propagation 166:176 propagation 166:176 propagation 166:176 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 1 18 theme potentials 188:197 arg1 initiation 151:160 initiation 151:160 initiation 151:160 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 3 19 theme immunoglobulin 444:457 arg1 β2 469:470 β2 469:470 β2 469:470 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 3 19 theme immunoglobulin 444:457 arg1 domain 459:464 The immunoglobulin domain 440:464 The immunoglobulin domain of β2 440:470 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 2 20 theme auxiliary 374:382 arg1 β2 393:394 β2 393:394 β2 393:394 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 20 theme auxiliary 374:382 arg1 subunit 384:390 an auxiliary subunit 371:390 an auxiliary subunit 371:390 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 5 21 theme molecular 747:755 arg1 basis 757:761 a molecular basis 745:761 a molecular basis for KIIIA specificity 745:783 Many interacting residues are specific to Nav1.2, revealing a molecular basis for KIIIA specificity. 30765605 4 22 theme extracellular 585:597 arg1 segments 599:606 the extracellular segments 581:606 the extracellular segments in repeats I to III 581:626 The 16-residue KIIIA interacts with the extracellular segments in repeats I to III, placing Lys7 at the entrance to the selectivity filter. 30765605 0 23 theme Na+ 43:45 arg1 channel 47:53 human Na+ channel Nav1.2 37:60 human Na+ channel Nav1.2 37:60 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 30765605 5 24 theme interacting 690:700 arg1 residues 702:709 Many interacting residues 685:709 Many interacting residues 685:709 Many interacting residues are specific to Nav1.2, revealing a molecular basis for KIIIA specificity. 30765605 1 25 dep initiation 151:160 arg1 the 147:149 the 147:149 the 147:149 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 1 26 theme voltage-gated 92:104 arg1 channel 113:119 The voltage-gated sodium channel Nav1.2 88:126 The voltage-gated sodium channel Nav1.2 88:126 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 1 26 theme voltage-gated 92:104 arg1 responsible 131:141 responsible 131:141 responsible 131:141 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 0 27 theme human 37:41 arg1 channel 47:53 human Na+ channel Nav1.2 37:60 human Na+ channel Nav1.2 37:60 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 30765605 6 28 theme Nav 881:883 arg1 channels 885:892 Nav channels 881:892 Nav channels 881:892 The structure establishes a framework for the rational design of subtype-specific blockers for Nav channels. 30765605 3 29 theme disulfide 529:537 arg1 bond 539:542 a disulfide bond 527:542 a disulfide bond 527:542 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 1 30 theme sodium 106:111 arg1 channel 113:119 The voltage-gated sodium channel Nav1.2 88:126 The voltage-gated sodium channel Nav1.2 88:126 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 1 30 theme sodium 106:111 arg1 responsible 131:141 responsible 131:141 responsible 131:141 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 2 31 theme cryo-electron 244:256 arg1 microscopy 258:267 the cryo-electron microscopy 240:267 the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms 240:437 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 32 theme pore 315:318 arg1 blocker 320:326 a peptidic pore blocker 304:326 a peptidic pore blocker 304:326 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 32 theme pore 315:318 arg1 KIIIA 345:349 the μ-conotoxin KIIIA 329:349 the μ-conotoxin KIIIA 329:349 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 1 33 theme central 206:212 arg1 system 222:227 the central nervous system 202:227 the central nervous system 202:227 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 4 34 dep III 624:626 arg1 to 621:622 to 621:622 to 621:622 The 16-residue KIIIA interacts with the extracellular segments in repeats I to III, placing Lys7 at the entrance to the selectivity filter. 30765605 1 35 from propagation 166:176 arg1 system 222:227 the central nervous system 202:227 the central nervous system 202:227 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 2 36 theme peptidic 306:313 arg1 blocker 320:326 a peptidic pore blocker 304:326 a peptidic pore blocker 304:326 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 2 36 theme peptidic 306:313 arg1 KIIIA 345:349 the μ-conotoxin KIIIA 329:349 the μ-conotoxin KIIIA 329:349 We report the cryo-electron microscopy structure of human Nav1.2 bound to a peptidic pore blocker, the μ-conotoxin KIIIA, in the presence of an auxiliary subunit, β2, to an overall resolution of 3.0 angstroms. 30765605 1 37 theme nervous 214:220 arg1 system 222:227 the central nervous system 202:227 the central nervous system 202:227 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 5 38 theme Many 685:688 arg1 residues 702:709 Many interacting residues 685:709 Many interacting residues 685:709 Many interacting residues are specific to Nav1.2, revealing a molecular basis for KIIIA specificity. 30765605 4 39 dep repeats 611:617 arg1 III 624:626 III 624:626 III 624:626 The 16-residue KIIIA interacts with the extracellular segments in repeats I to III, placing Lys7 at the entrance to the selectivity filter. 30765605 3 40 theme β2 469:470 arg1 β2 469:470 β2 469:470 β2 469:470 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 3 40 theme β2 469:470 arg1 domain 459:464 The immunoglobulin domain 440:464 The immunoglobulin domain of β2 440:470 The immunoglobulin domain of β2 interacts with the shoulder of the pore domain through a disulfide bond. 30765605 0 41 theme channel 47:53 arg1 blockade 25:32 pore blockade 20:32 pore blockade of human Na+ channel Nav1.2 20:60 Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. 30765605 6 42 theme blockers 868:875 arg1 design 841:846 the rational design 828:846 the rational design of subtype-specific blockers for Nav channels 828:892 The structure establishes a framework for the rational design of subtype-specific blockers for Nav channels. 30765605 1 43 from initiation 151:160 arg1 system 222:227 the central nervous system 202:227 the central nervous system 202:227 The voltage-gated sodium channel Nav1.2 is responsible for the initiation and propagation of action potentials in the central nervous system. 30765605 4 44 theme 16-residue 549:558 arg1 KIIIA 560:564 The 16-residue KIIIA 545:564 The 16-residue KIIIA 545:564 The 16-residue KIIIA interacts with the extracellular segments in repeats I to III, placing Lys7 at the entrance to the selectivity filter. 30559189 7 0 theme acidic 916:921 arg1 domain 923:928 GPIHBP1's acidic domain 906:928 GPIHBP1's acidic domain 906:928 GPIHBP1's acidic domain was not defined in the electron density map but was positioned to interact with LPL's large basic patch, providing a likely explanation for how GPIHBP1 stabilizes LPL. 30559189 4 1 theme recent 544:549 arg1 discovery 551:559 our recent discovery 540:559 our recent discovery that GPIHBP1's acidic domain preserves LPL structure and activity 540:625 Inspired by our recent discovery that GPIHBP1's acidic domain preserves LPL structure and activity, we crystallized an LPL-GPIHBP1 complex and solved its structure. 30559189 6 2 theme large 848:852 arg1 patch 860:864 a large basic patch 846:864 a large basic patch spanning its N- and C-terminal domains 846:903 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 2 3 theme LU 304:305 arg1 domain 307:312 a three-fingered LU domain 287:312 a three-fingered LU domain 287:312 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 2 4 theme N-terminal 321:330 arg1 domain 364:369 an N-terminal intrinsically disordered acidic domain 318:369 an N-terminal intrinsically disordered acidic domain 318:369 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 2 5 theme three-fingered 289:302 arg1 domain 307:312 a three-fingered LU domain 287:312 a three-fingered LU domain 287:312 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 1 6 theme triglyceride-rich 174:190 arg1 lipoproteins 192:203 triglyceride-rich lipoproteins 174:203 triglyceride-rich lipoproteins 174:203 Lipoprotein lipase (LPL) is responsible for the intravascular processing of triglyceride-rich lipoproteins. 30559189 3 7 theme Loss-of-function 372:387 arg1 mutations 389:397 Loss-of-function mutations 372:397 Loss-of-function mutations in LPL or GPIHBP1 372:415 Loss-of-function mutations in LPL or GPIHBP1 cause severe hypertriglyceridemia (chylomicronemia), but structures for LPL and GPIHBP1 have remained elusive. 30559189 7 8 theme electron 953:960 arg1 map 970:972 the electron density map 949:972 the electron density map 949:972 GPIHBP1's acidic domain was not defined in the electron density map but was positioned to interact with LPL's large basic patch, providing a likely explanation for how GPIHBP1 stabilizes LPL. 30559189 1 9 theme lipoproteins 192:203 arg1 processing 160:169 the intravascular processing 142:169 the intravascular processing of triglyceride-rich lipoproteins 142:203 Lipoprotein lipase (LPL) is responsible for the intravascular processing of triglyceride-rich lipoproteins. 30559189 7 10 theme density 962:968 arg1 map 970:972 the electron density map 949:972 the electron density map 949:972 GPIHBP1's acidic domain was not defined in the electron density map but was positioned to interact with LPL's large basic patch, providing a likely explanation for how GPIHBP1 stabilizes LPL. 30559189 1 11 theme Lipoprotein 98:108 arg1 lipase 110:115 Lipoprotein lipase 98:115 Lipoprotein lipase (LPL) 98:121 Lipoprotein lipase (LPL) is responsible for the intravascular processing of triglyceride-rich lipoproteins. 30559189 1 11 theme Lipoprotein 98:108 arg1 LPL 118:120 LPL 118:120 LPL 118:120 Lipoprotein lipase (LPL) is responsible for the intravascular processing of triglyceride-rich lipoproteins. 30559189 8 12 theme LPL-GPIHBP1 1102:1112 arg1 structure 1114:1122 The LPL-GPIHBP1 structure 1098:1122 The LPL-GPIHBP1 structure 1098:1122 The LPL-GPIHBP1 structure provides insights into mutations causing chylomicronemia. 30559189 6 13 theme surfaces 810:817 arg1 Analysis 784:791 Analysis 784:791 Analysis of electrostatic surfaces 784:817 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 0 14 theme lipoprotein 17:27 arg1 complex 44:50 the lipoprotein lipase-GPIHBP1 complex 13:50 the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis 13:95 Structure of the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis. 30559189 4 15 theme acidic 576:581 arg1 domain 583:588 GPIHBP1's acidic domain 566:588 GPIHBP1's acidic domain 566:588 Inspired by our recent discovery that GPIHBP1's acidic domain preserves LPL structure and activity, we crystallized an LPL-GPIHBP1 complex and solved its structure. 30559189 6 16 theme electrostatic 796:808 arg1 surfaces 810:817 electrostatic surfaces 796:817 electrostatic surfaces 796:817 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 4 17 theme LPL 600:602 arg1 structure 604:612 LPL structure 600:612 LPL structure 600:612 Inspired by our recent discovery that GPIHBP1's acidic domain preserves LPL structure and activity, we crystallized an LPL-GPIHBP1 complex and solved its structure. 30559189 0 18 theme complex 44:50 arg1 Structure 0:8 Structure 0:8 Structure of the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis 0:95 Structure of the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis. 30559189 5 19 theme C-terminal 728:737 arg1 domain 739:744 LPL's C-terminal domain 722:744 LPL's C-terminal domain 722:744 GPIHBP1's LU domain binds to LPL's C-terminal domain, largely by hydrophobic interactions. 30559189 6 20 theme C-terminal 886:895 arg1 domains 897:903 C-terminal domains 886:903 C-terminal domains 886:903 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 0 21 theme lipase-GPIHBP1 29:42 arg1 complex 44:50 the lipoprotein lipase-GPIHBP1 complex 13:50 the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis 13:95 Structure of the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis. 30559189 3 22 from mutations 389:397 arg1 GPIHBP1 409:415 GPIHBP1 409:415 GPIHBP1 409:415 Loss-of-function mutations in LPL or GPIHBP1 cause severe hypertriglyceridemia (chylomicronemia), but structures for LPL and GPIHBP1 have remained elusive. 30559189 3 22 from mutations 389:397 arg1 LPL 402:404 LPL 402:404 LPL 402:404 Loss-of-function mutations in LPL or GPIHBP1 cause severe hypertriglyceridemia (chylomicronemia), but structures for LPL and GPIHBP1 have remained elusive. 30559189 7 23 theme basic 1022:1026 arg1 patch 1028:1032 LPL's large basic patch 1010:1032 LPL's large basic patch 1010:1032 GPIHBP1's acidic domain was not defined in the electron density map but was positioned to interact with LPL's large basic patch, providing a likely explanation for how GPIHBP1 stabilizes LPL. 30559189 5 24 theme hydrophobic 758:768 arg1 interactions 770:781 hydrophobic interactions 758:781 hydrophobic interactions 758:781 GPIHBP1's LU domain binds to LPL's C-terminal domain, largely by hydrophobic interactions. 30559189 3 25 theme severe 423:428 arg1 hypertriglyceridemia 430:449 severe hypertriglyceridemia 423:449 severe hypertriglyceridemia (chylomicronemia) 423:467 Loss-of-function mutations in LPL or GPIHBP1 cause severe hypertriglyceridemia (chylomicronemia), but structures for LPL and GPIHBP1 have remained elusive. 30559189 3 25 theme severe 423:428 arg1 chylomicronemia 452:466 chylomicronemia 452:466 chylomicronemia 452:466 Loss-of-function mutations in LPL or GPIHBP1 cause severe hypertriglyceridemia (chylomicronemia), but structures for LPL and GPIHBP1 have remained elusive. 30559189 5 26 theme LU 703:704 arg1 domain 706:711 GPIHBP1's LU domain 693:711 GPIHBP1's LU domain 693:711 GPIHBP1's LU domain binds to LPL's C-terminal domain, largely by hydrophobic interactions. 30559189 6 27 contain contains 837:844 arg2 patch 860:864 a large basic patch 846:864 a large basic patch spanning its N- and C-terminal domains 846:903 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 6 27 contain contains 837:844 arg1 LPL 833:835 LPL 833:835 LPL 833:835 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 2 28 with protein 274:280 arg1 domain 364:369 an N-terminal intrinsically disordered acidic domain 318:369 an N-terminal intrinsically disordered acidic domain 318:369 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 2 28 with protein 274:280 arg1 domain 307:312 a three-fingered LU domain 287:312 a three-fingered LU domain 287:312 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 7 29 theme likely 1047:1052 arg1 explanation 1054:1064 a likely explanation 1045:1064 a likely explanation for how GPIHBP1 stabilizes LPL 1045:1095 GPIHBP1's acidic domain was not defined in the electron density map but was positioned to interact with LPL's large basic patch, providing a likely explanation for how GPIHBP1 stabilizes LPL. 30559189 2 30 theme acidic 357:362 arg1 domain 364:369 an N-terminal intrinsically disordered acidic domain 318:369 an N-terminal intrinsically disordered acidic domain 318:369 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 7 31 theme large 1016:1020 arg1 patch 1028:1032 LPL's large basic patch 1010:1032 LPL's large basic patch 1010:1032 GPIHBP1's acidic domain was not defined in the electron density map but was positioned to interact with LPL's large basic patch, providing a likely explanation for how GPIHBP1 stabilizes LPL. 30559189 2 32 theme disordered 346:355 arg1 domain 364:369 an N-terminal intrinsically disordered acidic domain 318:369 an N-terminal intrinsically disordered acidic domain 318:369 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 2 33 theme cell 269:272 arg1 GPIHBP1 245:251 GPIHBP1 245:251 GPIHBP1 245:251 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 2 33 theme cell 269:272 arg1 protein 274:280 an endothelial cell protein 254:280 an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain 254:369 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 0 34 theme triglyceride 73:84 arg1 hydrolysis 86:95 plasma triglyceride hydrolysis 66:95 plasma triglyceride hydrolysis 66:95 Structure of the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis. 30559189 6 35 theme basic 854:858 arg1 patch 860:864 a large basic patch 846:864 a large basic patch spanning its N- and C-terminal domains 846:903 Analysis of electrostatic surfaces revealed that LPL contains a large basic patch spanning its N- and C-terminal domains. 30559189 2 36 theme endothelial 257:267 arg1 GPIHBP1 245:251 GPIHBP1 245:251 GPIHBP1 245:251 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 2 36 theme endothelial 257:267 arg1 protein 274:280 an endothelial cell protein 254:280 an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain 254:369 The LPL within capillaries is bound to GPIHBP1, an endothelial cell protein with a three-fingered LU domain and an N-terminal intrinsically disordered acidic domain. 30559189 4 37 theme LPL-GPIHBP1 647:657 arg1 complex 659:665 an LPL-GPIHBP1 complex 644:665 an LPL-GPIHBP1 complex 644:665 Inspired by our recent discovery that GPIHBP1's acidic domain preserves LPL structure and activity, we crystallized an LPL-GPIHBP1 complex and solved its structure. 30559189 0 38 theme plasma 66:71 arg1 hydrolysis 86:95 plasma triglyceride hydrolysis 66:95 plasma triglyceride hydrolysis 66:95 Structure of the lipoprotein lipase-GPIHBP1 complex that mediates plasma triglyceride hydrolysis. 30559189 1 39 theme intravascular 146:158 arg1 processing 160:169 the intravascular processing 142:169 the intravascular processing of triglyceride-rich lipoproteins 142:203 Lipoprotein lipase (LPL) is responsible for the intravascular processing of triglyceride-rich lipoproteins. 30190309 6 0 theme drug 902:905 arg1 discovery 907:915 drug discovery 902:915 drug discovery 902:915 The structure provides a path toward mechanistic investigation of Nav channels and drug discovery for Nav channelopathies. 30190309 6 1 theme mechanistic 856:866 arg1 investigation 868:880 mechanistic investigation 856:880 mechanistic investigation of Nav channels 856:896 The structure provides a path toward mechanistic investigation of Nav channels and drug discovery for Nav channelopathies. 30190309 2 2 theme mechanistic 318:328 arg1 understanding 330:342 mechanistic understanding 318:342 mechanistic understanding 318:342 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 6 3 theme Nav 885:887 arg1 channels 889:896 Nav channels 885:896 Nav channels 885:896 The structure provides a path toward mechanistic investigation of Nav channels and drug discovery for Nav channelopathies. 30190309 4 4 theme β1 543:544 arg1 subunit 546:552 the β1 subunit 539:552 the β1 subunit 539:552 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 5 5 theme allosteric 750:759 arg1 mechanism 770:778 an allosteric blocking mechanism 747:778 an allosteric blocking mechanism for fast inactivation of Nav channels 747:816 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 5 6 theme reported 683:690 arg1 residues 703:710 reported functional residues 683:710 reported functional residues 683:710 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 0 7 from Structure 0:8 arg1 complex 62:68 complex 62:68 complex with β1 62:76 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 30190309 5 8 theme functional 692:701 arg1 residues 703:710 reported functional residues 683:710 reported functional residues 683:710 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 4 9 theme Accurate 452:459 arg1 building 467:474 Accurate model building 452:474 Accurate model building 452:474 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 0 10 with complex 62:68 arg1 β1 75:76 β1 75:76 β1 75:76 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 30190309 4 11 theme repeats 651:657 arg1 permeation 606:615 Na+ permeation 602:615 Na+ permeation 602:615 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 4 11 theme repeats 651:657 arg1 asymmetry 629:637 kinetic asymmetry 621:637 kinetic asymmetry 621:637 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 5 12 theme residues 703:710 arg1 analysis 671:678 Structural analysis 660:678 Structural analysis of reported functional residues and disease mutations 660:732 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 1 13 theme action 142:147 arg1 generation 159:168 action potential generation 142:168 action potential generation 142:168 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 0 14 theme channel 44:50 arg1 Structure 0:8 Structure 0:8 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 0:77 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 30190309 0 15 theme sodium 37:42 arg1 channel 44:50 the human voltage-gated sodium channel Nav1.4 13:57 the human voltage-gated sodium channel Nav1.4 13:57 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 30190309 1 16 theme potential 149:157 arg1 generation 159:168 action potential generation 142:168 action potential generation 142:168 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 5 17 theme fast 784:787 arg1 inactivation 789:800 fast inactivation 784:800 fast inactivation of Nav channels 784:816 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 4 18 theme model 461:465 arg1 building 467:474 Accurate model building 452:474 Accurate model building 452:474 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 1 19 theme Voltage-gated 79:91 arg1 channels 106:113 Voltage-gated sodium (Nav) channels 79:113 Voltage-gated sodium (Nav) channels 79:113 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 4 20 theme kinetic 621:627 arg1 asymmetry 629:637 kinetic asymmetry 621:637 kinetic asymmetry 621:637 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 0 21 theme human 17:21 arg1 channel 44:50 the human voltage-gated sodium channel Nav1.4 13:57 the human voltage-gated sodium channel Nav1.4 13:57 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 30190309 4 22 theme voltage-sensing 510:524 arg1 domains 526:532 the voltage-sensing domains 506:532 the voltage-sensing domains 506:532 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 2 23 theme structure 270:278 arg1 lack 260:263 the lack 256:263 the lack of a structure of any human Nav channel 256:303 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 3 24 theme 3.2-Å 434:438 arg1 resolution 440:449 3.2-Å resolution 434:449 3.2-Å resolution 434:449 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 5 25 theme Nav 805:807 arg1 channels 809:816 Nav channels 805:816 Nav channels 805:816 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 5 26 theme Structural 660:669 arg1 analysis 671:678 Structural analysis 660:678 Structural analysis of reported functional residues and disease mutations 660:732 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 4 27 theme molecular 582:590 arg1 basis 592:596 the molecular basis 578:596 the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats 578:657 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 3 28 theme microscopy 379:388 arg1 structure 390:398 the cryo-electron microscopy structure 361:398 the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution 361:449 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 4 29 theme Na+ 602:604 arg1 permeation 606:615 Na+ permeation 602:615 Na+ permeation 602:615 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 3 30 theme complex 423:429 arg1 structure 390:398 the cryo-electron microscopy structure 361:398 the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution 361:449 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 0 31 theme voltage-gated 23:35 arg1 channel 44:50 the human voltage-gated sodium channel Nav1.4 13:57 the human voltage-gated sodium channel Nav1.4 13:57 Structure of the human voltage-gated sodium channel Nav1.4 in complex with β1. 30190309 5 32 theme blocking 761:768 arg1 mechanism 770:778 an allosteric blocking mechanism 747:778 an allosteric blocking mechanism for fast inactivation of Nav channels 747:816 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 1 33 theme many 189:192 arg1 diseases 200:207 many human diseases 189:207 many human diseases 189:207 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 4 34 theme pore 493:496 arg1 domain 498:503 the pore domain 489:503 the pore domain 489:503 Accurate model building was made for the pore domain, the voltage-sensing domains, and the β1 subunit, providing insight into the molecular basis for Na+ permeation and kinetic asymmetry of the four repeats. 30190309 3 35 theme human 407:411 arg1 complex 423:429 the human Nav1.4-β1 complex 403:429 the human Nav1.4-β1 complex 403:429 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 1 36 theme Nav 101:103 arg1 channels 106:113 Voltage-gated sodium (Nav) channels 79:113 Voltage-gated sodium (Nav) channels 79:113 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 3 37 theme cryo-electron 365:377 arg1 structure 390:398 the cryo-electron microscopy structure 361:398 the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution 361:449 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 6 38 theme Nav 921:923 arg1 channelopathies 925:939 Nav channelopathies 921:939 Nav channelopathies 921:939 The structure provides a path toward mechanistic investigation of Nav channels and drug discovery for Nav channelopathies. 30190309 2 39 theme channel 297:303 arg1 structure 270:278 a structure 268:278 a structure of any human Nav channel 268:303 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 2 40 theme characterization 238:253 arg1 decades 218:224 decades 218:224 decades of rigorous characterization 218:253 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 3 41 from resolution 440:449 arg1 structure 390:398 the cryo-electron microscopy structure 361:398 the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution 361:449 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 2 42 theme Nav 293:295 arg1 channel 297:303 any human Nav channel 283:303 any human Nav channel 283:303 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 1 43 theme sodium 93:98 arg1 channels 106:113 Voltage-gated sodium (Nav) channels 79:113 Voltage-gated sodium (Nav) channels 79:113 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 1 44 theme human 194:198 arg1 diseases 200:207 many human diseases 189:207 many human diseases 189:207 Voltage-gated sodium (Nav) channels, which are responsible for action potential generation, are implicated in many human diseases. 30190309 2 45 theme rigorous 229:236 arg1 characterization 238:253 rigorous characterization 229:253 rigorous characterization 229:253 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 2 46 theme human 287:291 arg1 channel 297:303 any human Nav channel 283:303 any human Nav channel 283:303 Despite decades of rigorous characterization, the lack of a structure of any human Nav channel has hampered mechanistic understanding. 30190309 5 47 theme disease 716:722 arg1 mutations 724:732 disease mutations 716:732 disease mutations 716:732 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 5 48 theme channels 809:816 arg1 inactivation 789:800 fast inactivation 784:800 fast inactivation of Nav channels 784:816 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 30190309 3 49 theme Nav1.4-β1 413:421 arg1 complex 423:429 the human Nav1.4-β1 complex 403:429 the human Nav1.4-β1 complex 403:429 Here, we report the cryo-electron microscopy structure of the human Nav1.4-β1 complex at 3.2-Å resolution. 30190309 6 50 theme channels 889:896 arg1 investigation 868:880 mechanistic investigation 856:880 mechanistic investigation of Nav channels 856:896 The structure provides a path toward mechanistic investigation of Nav channels and drug discovery for Nav channelopathies. 30190309 6 50 theme channels 889:896 arg1 discovery 907:915 drug discovery 902:915 drug discovery 902:915 The structure provides a path toward mechanistic investigation of Nav channels and drug discovery for Nav channelopathies. 30190309 5 51 theme mutations 724:732 arg1 analysis 671:678 Structural analysis 660:678 Structural analysis of reported functional residues and disease mutations 660:732 Structural analysis of reported functional residues and disease mutations corroborates an allosteric blocking mechanism for fast inactivation of Nav channels. 24692546 6 0 theme antagonist 1007:1016 arg1 activities 1018:1027 the agonist and antagonist activities 991:1027 the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies 991:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 24692546 4 1 gly deglycosylated 650:663 arg1 FSH 665:667 deglycosylated FSH 650:667 deglycosylated FSH 650:667 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 6 2 theme autoantibodies 1069:1082 arg1 activities 1018:1027 the agonist and antagonist activities 991:1027 the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies 991:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 24692546 5 3 theme proposed 922:929 arg1 model 931:935 the proposed model 918:935 the proposed model 918:935 This study presents evidence confirming these predictions and provides crystallographic and mutagenesis data supporting the proposed model. 24692546 5 4 theme mutagenesis 890:900 arg1 data 902:905 crystallographic and mutagenesis data 869:905 crystallographic and mutagenesis data supporting the proposed model 869:935 This study presents evidence confirming these predictions and provides crystallographic and mutagenesis data supporting the proposed model. 24692546 4 5 theme FSHR 582:585 arg1 trimer 587:592 the FSHR trimer 578:592 the FSHR trimer 578:592 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 3 6 gly glycosylated 516:527 arg1 FSH 529:531 fully glycosylated FSH 510:531 fully glycosylated FSH 510:531 The model predicts that FSHR binds Asnα(52)-deglycosylated FSH at a 3-fold higher capacity than fully glycosylated FSH. 24692546 3 7 theme Asnα 449:452 arg1 FSH 473:475 Asnα(52)-deglycosylated FSH 449:475 Asnα(52)-deglycosylated FSH 449:475 The model predicts that FSHR binds Asnα(52)-deglycosylated FSH at a 3-fold higher capacity than fully glycosylated FSH. 24692546 1 8 theme Follicle-stimulating 75:94 arg1 target 172:177 an important drug target 154:177 an important drug target in the development of novel therapeutics for reproductive indications 154:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 8 theme Follicle-stimulating 75:94 arg1 FSHR 114:117 FSHR 114:117 FSHR 114:117 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 8 theme Follicle-stimulating 75:94 arg1 receptor 141:148 a G-protein coupled receptor 121:148 a G-protein coupled receptor 121:148 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 8 theme Follicle-stimulating 75:94 arg1 receptor 104:111 Follicle-stimulating hormone receptor 75:111 Follicle-stimulating hormone receptor (FSHR) 75:118 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 6 9 theme thyroid-stimulating 1032:1050 arg1 receptor 1060:1067 thyroid-stimulating hormone receptor 1032:1067 thyroid-stimulating hormone receptor autoantibodies 1032:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 24692546 1 10 theme important 157:165 arg1 target 172:177 an important drug target 154:177 an important drug target in the development of novel therapeutics for reproductive indications 154:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 10 theme important 157:165 arg1 receptor 104:111 Follicle-stimulating hormone receptor 75:111 Follicle-stimulating hormone receptor (FSHR) 75:118 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 4 11 theme signaling 766:774 arg1 activities 776:785 signaling activities 766:785 signaling activities 766:785 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 1 12 theme hormone 96:102 arg1 target 172:177 an important drug target 154:177 an important drug target in the development of novel therapeutics for reproductive indications 154:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 12 theme hormone 96:102 arg1 FSHR 114:117 FSHR 114:117 FSHR 114:117 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 12 theme hormone 96:102 arg1 receptor 141:148 a G-protein coupled receptor 121:148 a G-protein coupled receptor 121:148 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 12 theme hormone 96:102 arg1 receptor 104:111 Follicle-stimulating hormone receptor 75:111 Follicle-stimulating hormone receptor (FSHR) 75:118 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 13 theme drug 167:170 arg1 target 172:177 an important drug target 154:177 an important drug target in the development of novel therapeutics for reproductive indications 154:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 13 theme drug 167:170 arg1 receptor 104:111 Follicle-stimulating hormone receptor 75:111 Follicle-stimulating hormone receptor (FSHR) 75:118 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 3 14 gly -deglycosylated 457:471 arg1 FSH 473:475 Asnα(52)-deglycosylated FSH 449:475 Asnα(52)-deglycosylated FSH 449:475 The model predicts that FSHR binds Asnα(52)-deglycosylated FSH at a 3-fold higher capacity than fully glycosylated FSH. 24692546 2 15 theme novel 363:367 arg1 model 369:373 a novel model 361:373 a novel model for the receptor activation mechanism 361:411 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 4 16 theme FSH 665:667 arg1 binding 613:619 the binding 609:619 the binding of glycosylated FSH, but not deglycosylated FSH, 609:668 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 0 17 theme Hormone 34:40 arg1 Receptor 42:49 Follicle-stimulating Hormone Receptor 13:49 Follicle-stimulating Hormone Receptor as a Functional Trimer 13:72 Evidence for Follicle-stimulating Hormone Receptor as a Functional Trimer. 24692546 2 18 theme crystal 302:308 arg1 structure 310:318 the crystal structure 298:318 the crystal structure 298:318 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 4 19 theme trimer 587:592 arg1 dissociation 562:573 dissociation 562:573 dissociation of the FSHR trimer into monomers 562:606 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 0 20 theme Follicle-stimulating 13:32 arg1 Receptor 42:49 Follicle-stimulating Hormone Receptor 13:49 Follicle-stimulating Hormone Receptor as a Functional Trimer 13:72 Evidence for Follicle-stimulating Hormone Receptor as a Functional Trimer. 24692546 3 21 theme higher 489:494 arg1 capacity 496:503 a 3-fold higher capacity 480:503 a 3-fold higher capacity than fully glycosylated FSH 480:531 The model predicts that FSHR binds Asnα(52)-deglycosylated FSH at a 3-fold higher capacity than fully glycosylated FSH. 24692546 2 22 theme activation 392:401 arg1 mechanism 403:411 the receptor activation mechanism 379:411 the receptor activation mechanism 379:411 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 4 23 theme FSHR 749:752 arg1 binding 754:760 FSHR binding 749:760 FSHR binding 749:760 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 2 24 theme receptor 383:390 arg1 mechanism 403:411 the receptor activation mechanism 379:411 the receptor activation mechanism 379:411 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 6 25 theme receptor 1060:1067 arg1 autoantibodies 1069:1082 thyroid-stimulating hormone receptor autoantibodies 1032:1082 thyroid-stimulating hormone receptor autoantibodies 1032:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 24692546 2 26 located observed 286:293 arg2 domains 273:279 The FSHR extracellular domains 250:279 The FSHR extracellular domains 250:279 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 2 26 located observed 286:293 arg1 structure 310:318 the crystal structure 298:318 the crystal structure 298:318 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 2 26 located observed 286:293 arg2 trimer 325:330 a trimer 323:330 a trimer 323:330 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 1 27 theme novel 201:205 arg1 therapeutics 207:218 novel therapeutics 201:218 novel therapeutics for reproductive indications 201:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 0 28 theme Functional 56:65 arg1 Trimer 67:72 a Functional Trimer 54:72 a Functional Trimer 54:72 Evidence for Follicle-stimulating Hormone Receptor as a Functional Trimer. 24692546 4 29 gly glycosylated 624:635 arg1 FSH 637:639 glycosylated FSH 624:639 glycosylated FSH 624:639 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 4 30 theme deglycosylated 650:663 arg1 FSH 665:667 deglycosylated FSH 650:667 deglycosylated FSH 650:667 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 7 31 theme functional 1119:1128 arg1 trimer 1130:1135 a functional trimer 1117:1135 a functional trimer 1117:1135 We conclude that FSHR exists as a functional trimer. 24692546 3 32 theme -deglycosylated 457:471 arg1 FSH 473:475 Asnα(52)-deglycosylated FSH 449:475 Asnα(52)-deglycosylated FSH 449:475 The model predicts that FSHR binds Asnα(52)-deglycosylated FSH at a 3-fold higher capacity than fully glycosylated FSH. 24692546 2 33 theme extracellular 259:271 arg1 domains 273:279 The FSHR extracellular domains 250:279 The FSHR extracellular domains 250:279 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 2 33 theme extracellular 259:271 arg1 trimer 325:330 a trimer 323:330 a trimer 323:330 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 1 34 theme G-protein 123:131 arg1 receptor 104:111 Follicle-stimulating hormone receptor 75:111 Follicle-stimulating hormone receptor (FSHR) 75:118 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 34 theme G-protein 123:131 arg1 receptor 141:148 a G-protein coupled receptor 121:148 a G-protein coupled receptor 121:148 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 3 35 theme glycosylated 516:527 arg1 FSH 529:531 fully glycosylated FSH 510:531 fully glycosylated FSH 510:531 The model predicts that FSHR binds Asnα(52)-deglycosylated FSH at a 3-fold higher capacity than fully glycosylated FSH. 24692546 1 36 theme therapeutics 207:218 arg1 development 186:196 the development 182:196 the development of novel therapeutics for reproductive indications 182:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 4 37 theme dissociated 706:716 arg1 monomers 718:725 the dissociated monomers 702:725 the dissociated monomers 702:725 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 2 38 theme FSHR 254:257 arg1 domains 273:279 The FSHR extracellular domains 250:279 The FSHR extracellular domains 250:279 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 2 38 theme FSHR 254:257 arg1 trimer 325:330 a trimer 323:330 a trimer 323:330 The FSHR extracellular domains were observed in the crystal structure as a trimer, which enabled us to propose a novel model for the receptor activation mechanism. 24692546 1 39 from target 172:177 arg1 development 186:196 the development 182:196 the development of novel therapeutics for reproductive indications 182:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 40 theme coupled 133:139 arg1 receptor 104:111 Follicle-stimulating hormone receptor 75:111 Follicle-stimulating hormone receptor (FSHR) 75:118 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 1 40 theme coupled 133:139 arg1 receptor 141:148 a G-protein coupled receptor 121:148 a G-protein coupled receptor 121:148 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 5 41 theme crystallographic 869:884 arg1 data 902:905 crystallographic and mutagenesis data 869:905 crystallographic and mutagenesis data supporting the proposed model 869:935 This study presents evidence confirming these predictions and provides crystallographic and mutagenesis data supporting the proposed model. 24692546 6 42 theme agonist 995:1001 arg1 activities 1018:1027 the agonist and antagonist activities 991:1027 the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies 991:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 24692546 4 43 theme FSH 637:639 arg1 binding 613:619 the binding 609:619 the binding of glycosylated FSH, but not deglycosylated FSH, 609:668 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 6 44 theme hormone 1052:1058 arg1 receptor 1060:1067 thyroid-stimulating hormone receptor 1032:1067 thyroid-stimulating hormone receptor autoantibodies 1032:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 24692546 1 45 theme reproductive 224:235 arg1 indications 237:247 reproductive indications 224:247 reproductive indications 224:247 Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor, is an important drug target in the development of novel therapeutics for reproductive indications. 24692546 4 46 theme glycosylated 624:635 arg1 FSH 637:639 glycosylated FSH 624:639 glycosylated FSH 624:639 It also predicts that, upon dissociation of the FSHR trimer into monomers, the binding of glycosylated FSH, but not deglycosylated FSH, would increase 3-fold, and that the dissociated monomers would in turn enhance FSHR binding and signaling activities by 3-fold. 24692546 6 47 theme mechanistic 964:974 arg1 explanation 976:986 a mechanistic explanation 962:986 a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies 962:1082 The model also provides a mechanistic explanation to the agonist and antagonist activities of thyroid-stimulating hormone receptor autoantibodies. 20033057 7 0 theme extension 1056:1064 arg1 role 1048:1051 the critical role 1035:1051 the critical role of extension 1035:1064 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 7 0 theme extension 1056:1064 arg1 conversion 1071:1080 conversion 1071:1080 conversion from the closed to the open headpiece conformation 1071:1131 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 8 1 from information 1231:1241 arg1 flexibility 1258:1268 interdomain flexibility 1246:1268 interdomain flexibility 1246:1268 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 20033057 1 2 theme complement 159:168 arg1 domain 130:135 an alphaI domain 120:135 an alphaI domain 120:135 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 2 theme complement 159:168 arg1 type 179:182 the complement receptor type 4 155:184 the complement receptor type 4 155:184 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 0 3 theme receptor 59:66 arg1 type 68:71 complement receptor type 4 48:73 complement receptor type 4 48:73 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 0 3 theme receptor 59:66 arg1 domain 40:45 an alphaI domain 30:45 an alphaI domain 30:45 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 1 4 theme alphaI 123:128 arg1 type 179:182 the complement receptor type 4 155:184 the complement receptor type 4 155:184 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 4 theme alphaI 123:128 arg1 domain 130:135 an alphaI domain 120:135 an alphaI domain 120:135 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 4 theme alphaI 123:128 arg1 beta 146:149 alpha(X)beta(2) 138:152 alpha(X)beta(2) 138:152 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 5 theme receptor 170:177 arg1 domain 130:135 an alphaI domain 120:135 an alphaI domain 120:135 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 5 theme receptor 170:177 arg1 type 179:182 the complement receptor type 4 155:184 the complement receptor type 4 155:184 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 3 6 theme domain 436:441 arg1 states 458:463 two betaI domain conformational states 426:463 two betaI domain conformational states 426:463 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 6 7 theme beta 857:860 arg1 domain 870:875 the beta(2)-tail domain 853:875 the beta(2)-tail domain 853:875 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 5 8 theme transmembrane 791:803 arg1 domains 805:811 the transmembrane domains 787:811 the transmembrane domains 787:811 Linkers extending to the transmembrane domains are unstructured. 20033057 4 9 theme alphaI 622:627 arg1 domains 629:635 alphaI domains 622:635 alphaI domains 622:635 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 7 10 from closed 1091:1096 arg1 role 1048:1051 the critical role 1035:1051 the critical role of extension 1035:1064 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 7 10 from closed 1091:1096 arg1 conversion 1071:1080 conversion 1071:1080 conversion from the closed to the open headpiece conformation 1071:1131 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 3 11 theme conformational 443:456 arg1 states 458:463 two betaI domain conformational states 426:463 two betaI domain conformational states 426:463 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 7 12 theme open 1105:1108 arg1 conformation 1120:1131 the open headpiece conformation 1101:1131 the open headpiece conformation 1101:1131 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 7 13 theme headpiece 1110:1118 arg1 conformation 1120:1131 the open headpiece conformation 1101:1131 the open headpiece conformation 1101:1131 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 4 14 theme X 571:571 arg1 2 578:578 2 578:578 2 578:578 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 14 theme X 571:571 arg1 beta 573:576 alpha(X)beta 565:576 alpha(X)beta(2) 565:579 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 15 theme alpha 565:569 arg1 2 578:578 2 578:578 2 578:578 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 15 theme alpha 565:569 arg1 beta 573:576 alpha(X)beta 565:576 alpha(X)beta(2) 565:579 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 7 16 theme critical 1039:1046 arg1 role 1048:1051 the critical role 1035:1051 the critical role of extension 1035:1064 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 6 17 from importance 889:898 arg1 activation 950:959 integrin activation 941:959 integrin activation 941:959 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 3 18 theme betaI 430:434 arg1 states 458:463 two betaI domain conformational states 426:463 two betaI domain conformational states 426:463 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 2 19 theme signal 349:354 arg1 transmission 356:367 allosteric signal transmission 338:367 allosteric signal transmission 338:367 It was earlier expected that a fixed orientation between the alphaI domain and the beta-propeller domain in which it is inserted would be required for allosteric signal transmission. 20033057 2 20 theme fixed 218:222 arg1 orientation 224:234 a fixed orientation 216:234 a fixed orientation between the alphaI domain and the beta-propeller domain in which it is inserted 216:314 It was earlier expected that a fixed orientation between the alphaI domain and the beta-propeller domain in which it is inserted would be required for allosteric signal transmission. 20033057 1 21 theme alpha 138:142 arg1 domain 130:135 an alphaI domain 120:135 an alphaI domain 120:135 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 21 theme alpha 138:142 arg1 beta 146:149 alpha(X)beta(2) 138:152 alpha(X)beta(2) 138:152 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 2 22 theme allosteric 338:347 arg1 transmission 356:367 allosteric signal transmission 338:367 allosteric signal transmission 338:367 It was earlier expected that a fixed orientation between the alphaI domain and the beta-propeller domain in which it is inserted would be required for allosteric signal transmission. 20033057 1 23 with structure 90:98 arg1 type 179:182 the complement receptor type 4 155:184 the complement receptor type 4 155:184 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 23 with structure 90:98 arg1 domain 130:135 an alphaI domain 120:135 an alphaI domain 120:135 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 23 with structure 90:98 arg1 beta 146:149 alpha(X)beta(2) 138:152 alpha(X)beta(2) 138:152 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 8 24 from Differences 1158:1168 arg1 lattices 1200:1207 crystal lattices 1192:1207 crystal lattices 1192:1207 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 20033057 0 25 theme integrin 16:23 arg1 Structure 0:8 Structure 0:8 Structure of an integrin with an alphaI domain, complement receptor type 4. 0:74 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 4 26 theme beta-legs 677:685 arg1 beta-legs 677:685 beta-legs 677:685 beta-legs 677:685 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 26 theme beta-legs 677:685 arg1 domains 651:657 the terminal domains 638:657 the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, 638:708 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 26 theme beta-legs 677:685 arg1 alpha- 666:671 alpha- 666:671 alpha- 666:671 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 1 27 theme X 144:144 arg1 domain 130:135 an alphaI domain 120:135 an alphaI domain 120:135 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 1 27 theme X 144:144 arg1 beta 146:149 alpha(X)beta(2) 138:152 alpha(X)beta(2) 138:152 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 7 28 theme antibody 1012:1019 arg1 epitopes 1021:1028 antibody epitopes 1012:1028 antibody epitopes 1012:1028 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 6 29 theme Previous 831:838 arg1 mutations 840:848 Previous mutations 831:848 Previous mutations in the beta(2)-tail domain 831:875 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 8 30 theme interdomain 1246:1256 arg1 flexibility 1258:1268 interdomain flexibility 1246:1268 interdomain flexibility 1246:1268 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 20033057 6 31 theme deadbolt 928:935 arg1 importance 889:898 the importance 885:898 the importance of extension, rather than a deadbolt, in integrin activation 885:959 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 8 32 theme unprecedented 1217:1229 arg1 information 1231:1241 unprecedented information 1217:1241 unprecedented information on interdomain flexibility important for modelling integrin extension and activation 1217:1326 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 20033057 1 33 theme integrin 106:113 arg1 structure 90:98 the structure 86:98 the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4 86:184 We report the structure of an integrin with an alphaI domain, alpha(X)beta(2), the complement receptor type 4. 20033057 7 34 theme activating 987:996 arg1 mutations 998:1006 further activating mutations 979:1006 further activating mutations 979:1006 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 3 35 theme domain 491:496 arg1 states 498:503 three alphaI domain states 478:503 three alphaI domain states 478:503 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 7 36 theme mutations 998:1006 arg1 locations 966:974 The locations 962:974 The locations of further activating mutations and antibody epitopes 962:1028 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 6 37 from mutations 840:848 arg1 domain 870:875 the beta(2)-tail domain 853:875 the beta(2)-tail domain 853:875 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 4 38 theme alpha- 666:671 arg1 beta-legs 677:685 beta-legs 677:685 beta-legs 677:685 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 38 theme alpha- 666:671 arg1 domains 651:657 the terminal domains 638:657 the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, 638:708 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 38 theme alpha- 666:671 arg1 alpha- 666:671 alpha- 666:671 alpha- 666:671 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 39 theme alphaI-less 743:753 arg1 integrins 755:763 alphaI-less integrins 743:763 alphaI-less integrins 743:763 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 3 40 theme ligand 536:541 arg1 recognition 543:553 ligand recognition 536:553 ligand recognition 536:553 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 6 41 theme integrin 941:948 arg1 activation 950:959 integrin activation 941:959 integrin activation 941:959 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 2 42 theme alphaI 248:253 arg1 domain 255:260 the alphaI domain 244:260 the alphaI domain 244:260 It was earlier expected that a fixed orientation between the alphaI domain and the beta-propeller domain in which it is inserted would be required for allosteric signal transmission. 20033057 0 43 theme alphaI 33:38 arg1 type 68:71 complement receptor type 4 48:73 complement receptor type 4 48:73 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 0 43 theme alphaI 33:38 arg1 domain 40:45 an alphaI domain 30:45 an alphaI domain 30:45 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 3 44 theme alphaI 484:489 arg1 states 498:503 three alphaI domain states 478:503 three alphaI domain states 478:503 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 7 45 theme further 979:985 arg1 mutations 998:1006 further activating mutations 979:1006 further activating mutations 979:1006 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 8 46 theme integrin 1294:1301 arg1 extension 1303:1311 integrin extension 1294:1311 integrin extension 1294:1311 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 20033057 3 47 theme greater 510:516 arg1 accessibility 518:530 greater accessibility 510:530 greater accessibility for ligand recognition 510:553 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 0 48 with Structure 0:8 arg1 type 68:71 complement receptor type 4 48:73 complement receptor type 4 48:73 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 0 48 with Structure 0:8 arg1 domain 40:45 an alphaI domain 30:45 an alphaI domain 30:45 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 8 49 theme crystal 1192:1198 arg1 lattices 1200:1207 crystal lattices 1192:1207 crystal lattices 1192:1207 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 20033057 4 50 theme terminal 642:649 arg1 beta-legs 677:685 beta-legs 677:685 beta-legs 677:685 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 50 theme terminal 642:649 arg1 domains 651:657 the terminal domains 638:657 the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, 638:708 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 4 50 theme terminal 642:649 arg1 alpha- 666:671 alpha- 666:671 alpha- 666:671 Although alpha(X)beta(2) is bent similarly to integrins that lack alphaI domains, the terminal domains of the alpha- and beta-legs, calf-2 and beta-tail, are oriented differently than in alphaI-less integrins. 20033057 3 51 theme alphaI 383:388 arg1 domain 390:395 the alphaI domain 379:395 the alphaI domain 379:395 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 3 51 theme alphaI 383:388 arg1 flexible 407:414 flexible 407:414 flexible 407:414 However, the alphaI domain is highly flexible, enabling two betaI domain conformational states to couple to three alphaI domain states, and greater accessibility for ligand recognition. 20033057 0 52 theme complement 48:57 arg1 type 68:71 complement receptor type 4 48:73 complement receptor type 4 48:73 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 0 52 theme complement 48:57 arg1 domain 40:45 an alphaI domain 30:45 an alphaI domain 30:45 Structure of an integrin with an alphaI domain, complement receptor type 4. 20033057 6 53 theme extension 903:911 arg1 importance 889:898 the importance 885:898 the importance of extension, rather than a deadbolt, in integrin activation 885:959 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 2 54 theme beta-propeller 270:283 arg1 domain 285:290 the beta-propeller domain 266:290 the beta-propeller domain 266:290 It was earlier expected that a fixed orientation between the alphaI domain and the beta-propeller domain in which it is inserted would be required for allosteric signal transmission. 20033057 7 55 theme integrin 1137:1144 arg1 activation 1146:1155 integrin activation 1137:1155 integrin activation 1137:1155 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 6 56 theme -tail 864:868 arg1 domain 870:875 the beta(2)-tail domain 853:875 the beta(2)-tail domain 853:875 Previous mutations in the beta(2)-tail domain support the importance of extension, rather than a deadbolt, in integrin activation. 20033057 7 57 theme epitopes 1021:1028 arg1 locations 966:974 The locations 962:974 The locations of further activating mutations and antibody epitopes 962:1028 The locations of further activating mutations and antibody epitopes show the critical role of extension, and conversion from the closed to the open headpiece conformation, in integrin activation. 20033057 8 58 theme important 1270:1278 arg1 information 1231:1241 unprecedented information 1217:1241 unprecedented information on interdomain flexibility important for modelling integrin extension and activation 1217:1326 Differences among 10 molecules in crystal lattices provide unprecedented information on interdomain flexibility important for modelling integrin extension and activation. 10871631 9 0 theme states 1658:1663 arg1 forms 1580:1584 torsion dystonia-associated forms 1552:1584 torsion dystonia-associated forms of torsin A 1552:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 0 theme states 1658:1663 arg1 proteins 1627:1634 properly folded, lumenal proteins 1602:1634 properly folded, lumenal proteins of similar oligomeric states 1602:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 0 theme states 1658:1663 arg1 type 1543:1546 the wild type 1534:1546 the wild type 1534:1546 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 6 1 theme resident 1069:1076 arg1 BiP 1086:1088 the endoplasmic reticulum resident protein BiP 1043:1088 the endoplasmic reticulum resident protein BiP 1043:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 10 2 theme A 1755:1755 arg1 phenotype 1782:1790 the disease-inducing phenotype 1761:1790 the disease-inducing phenotype of the protein 1761:1805 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 10 2 theme A 1755:1755 arg1 distribution 1725:1736 the altered subcellular distribution 1701:1736 the altered subcellular distribution of DeltaE-torsin A 1701:1755 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 7 3 theme DeltaE-torsin 1240:1252 arg1 A 1254:1254 DeltaE-torsin A 1240:1254 DeltaE-torsin A 1240:1254 These results initially suggested that DeltaE-torsin A might exist as insoluble aggregates. 10871631 1 4 theme DYT1 281:284 arg1 gene 286:289 the DYT1 gene 277:289 the DYT1 gene 277:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 3 5 theme one 514:516 arg1 loss 506:509 the loss 502:509 the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A) 502:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 1 6 theme movement 200:207 arg1 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 6 theme movement 200:207 arg1 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 6 7 theme spheroid 1132:1139 arg1 structures 1155:1164 large spheroid intracellular structures 1126:1164 large spheroid intracellular structures exclusive of BiP immunoreactivity 1126:1198 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 3 8 theme acid 540:543 arg1 residues 545:552 glutamic acid residues 531:552 glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A) 531:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 9 9 theme torsin 1589:1594 arg1 A 1596:1596 torsin A 1589:1596 torsin A 1589:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 5 10 theme oriented 864:871 arg1 glycoproteins 873:885 lumenally oriented glycoproteins 854:885 lumenally oriented glycoproteins 854:885 When transfected into mammalian cells, both torsin A and DeltaE-torsin A were found to behave as lumenally oriented glycoproteins. 10871631 6 11 theme Immunofluorescence 888:905 arg1 studies 907:913 Immunofluorescence studies 888:913 Immunofluorescence studies 888:913 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 8 12 theme migration 1450:1458 arg1 patterns 1460:1467 equivalent migration patterns 1439:1467 equivalent migration patterns 1439:1467 However, both torsin A and DeltaE-torsin A were readily solubilized by nonionic detergents, were similarly accessible to proteases, and displayed equivalent migration patterns on sucrose gradients. 10871631 10 13 theme DeltaE-torsin 1741:1753 arg1 A 1755:1755 DeltaE-torsin A 1741:1755 DeltaE-torsin A 1741:1755 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 8 14 theme nonionic 1364:1371 arg1 detergents 1373:1382 nonionic detergents 1364:1382 nonionic detergents 1364:1382 However, both torsin A and DeltaE-torsin A were readily solubilized by nonionic detergents, were similarly accessible to proteases, and displayed equivalent migration patterns on sucrose gradients. 10871631 1 15 theme Early-onset 133:143 arg1 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 15 theme Early-onset 133:143 arg1 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 8 16 theme torsin 1307:1312 arg1 A 1314:1314 torsin A 1307:1314 torsin A 1307:1314 However, both torsin A and DeltaE-torsin A were readily solubilized by nonionic detergents, were similarly accessible to proteases, and displayed equivalent migration patterns on sucrose gradients. 10871631 3 17 theme torsin 586:591 arg1 A 593:593 torsin A 586:593 torsin A (DeltaE-torsin A) 586:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 3 17 theme torsin 586:591 arg1 A 610:610 DeltaE-torsin A 596:610 DeltaE-torsin A 596:610 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 6 18 theme immunoreactivity 1183:1198 arg1 exclusive 1166:1174 exclusive 1166:1174 exclusive 1166:1174 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 5 19 theme DeltaE-torsin 814:826 arg1 A 828:828 DeltaE-torsin A 814:828 DeltaE-torsin A 814:828 When transfected into mammalian cells, both torsin A and DeltaE-torsin A were found to behave as lumenally oriented glycoproteins. 10871631 1 20 theme dominant 178:185 arg1 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 20 theme dominant 178:185 arg1 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 9 21 theme torsion 1552:1558 arg1 forms 1580:1584 torsion dystonia-associated forms 1552:1584 torsion dystonia-associated forms of torsin A 1552:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 21 theme torsion 1552:1558 arg1 proteins 1627:1634 properly folded, lumenal proteins 1602:1634 properly folded, lumenal proteins of similar oligomeric states 1602:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 21 theme torsion 1552:1558 arg1 type 1543:1546 the wild type 1534:1546 the wild type 1534:1546 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 6 22 theme torsin 929:934 arg1 A 936:936 torsin A 929:936 torsin A 929:936 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 10 23 theme potential 1670:1678 arg1 relationship 1680:1691 The potential relationship 1666:1691 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein 1666:1805 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 0 24 theme subcellular 106:116 arg1 localizations 118:130 distinct subcellular localizations 97:130 distinct subcellular localizations 97:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 9 25 theme wild 1538:1541 arg1 forms 1580:1584 torsion dystonia-associated forms 1552:1584 torsion dystonia-associated forms of torsin A 1552:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 25 theme wild 1538:1541 arg1 proteins 1627:1634 properly folded, lumenal proteins 1602:1634 properly folded, lumenal proteins of similar oligomeric states 1602:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 25 theme wild 1538:1541 arg1 type 1543:1546 the wild type 1534:1546 the wild type 1534:1546 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 0 26 theme Torsin 0:5 arg1 glycoproteins 70:82 lumenal glycoproteins 62:82 lumenal glycoproteins that exhibit distinct subcellular localizations 62:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 26 theme Torsin 0:5 arg1 A 7:7 Torsin A 0:7 Torsin A 0:7 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 26 theme Torsin 0:5 arg1 forms 52:56 its torsion dystonia-associated mutant forms 13:56 its torsion dystonia-associated mutant forms 13:56 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 4 27 dep expression 652:661 arg1 the 648:650 the 648:650 the 648:650 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 2 28 theme ATPase 418:423 arg1 chaperones 425:434 ATPase chaperones 418:434 ATPase chaperones 418:434 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 10 29 theme altered 1705:1711 arg1 distribution 1725:1736 the altered subcellular distribution 1701:1736 the altered subcellular distribution of DeltaE-torsin A 1701:1755 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 4 30 theme protein 748:754 arg1 expression 652:661 expression 652:661 expression 652:661 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 0 31 theme torsion 17:23 arg1 glycoproteins 70:82 lumenal glycoproteins 62:82 lumenal glycoproteins that exhibit distinct subcellular localizations 62:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 31 theme torsion 17:23 arg1 A 7:7 Torsin A 0:7 Torsin A 0:7 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 31 theme torsion 17:23 arg1 forms 52:56 its torsion dystonia-associated mutant forms 13:56 its torsion dystonia-associated mutant forms 13:56 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 9 32 theme folded 1611:1616 arg1 forms 1580:1584 torsion dystonia-associated forms 1552:1584 torsion dystonia-associated forms of torsin A 1552:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 32 theme folded 1611:1616 arg1 proteins 1627:1634 properly folded, lumenal proteins 1602:1634 properly folded, lumenal proteins of similar oligomeric states 1602:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 32 theme folded 1611:1616 arg1 type 1543:1546 the wild type 1534:1546 the wild type 1534:1546 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 6 33 theme intracellular 972:984 arg1 membranes 986:994 intracellular membranes 972:994 intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP 972:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 3 34 from deletion 441:448 arg1 DYT1 453:456 DYT1 453:456 DYT1 associated with torsion dystonia 453:489 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 3 35 theme torsion 474:480 arg1 dystonia 482:489 torsion dystonia 474:489 torsion dystonia 474:489 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 0 36 theme mutant 45:50 arg1 glycoproteins 70:82 lumenal glycoproteins 62:82 lumenal glycoproteins that exhibit distinct subcellular localizations 62:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 36 theme mutant 45:50 arg1 A 7:7 Torsin A 0:7 Torsin A 0:7 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 36 theme mutant 45:50 arg1 forms 52:56 its torsion dystonia-associated mutant forms 13:56 its torsion dystonia-associated mutant forms 13:56 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 6 37 theme DeltaE-torsin 1099:1111 arg1 A 1113:1113 DeltaE-torsin A 1099:1113 DeltaE-torsin A 1099:1113 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 4 38 theme A 727:727 arg1 expression 652:661 expression 652:661 expression 652:661 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 7 39 theme insoluble 1271:1279 arg1 aggregates 1281:1290 insoluble aggregates 1271:1290 insoluble aggregates 1271:1290 These results initially suggested that DeltaE-torsin A might exist as insoluble aggregates. 10871631 8 40 theme equivalent 1439:1448 arg1 patterns 1460:1467 equivalent migration patterns 1439:1467 equivalent migration patterns 1439:1467 However, both torsin A and DeltaE-torsin A were readily solubilized by nonionic detergents, were similarly accessible to proteases, and displayed equivalent migration patterns on sucrose gradients. 10871631 10 41 theme disease-inducing 1765:1780 arg1 phenotype 1782:1790 the disease-inducing phenotype 1761:1790 the disease-inducing phenotype of the protein 1761:1805 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 1 42 theme 3-base 253:258 arg1 deletion 265:272 a 3-base pair deletion 251:272 a 3-base pair deletion in the DYT1 gene 251:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 9 43 theme oligomeric 1647:1656 arg1 states 1658:1663 similar oligomeric states 1639:1663 similar oligomeric states 1639:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 6 44 theme protein 1078:1084 arg1 BiP 1086:1088 the endoplasmic reticulum resident protein BiP 1043:1088 the endoplasmic reticulum resident protein BiP 1043:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 6 45 theme reticulum 1059:1067 arg1 BiP 1086:1088 the endoplasmic reticulum resident protein BiP 1043:1088 the endoplasmic reticulum resident protein BiP 1043:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 2 46 theme DYT1 296:299 arg1 protein 331:337 a 332-amino acid protein 314:337 a 332-amino acid protein 314:337 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 46 theme DYT1 296:299 arg1 gene 301:304 The DYT1 gene 292:304 The DYT1 gene 292:304 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 47 theme 332-amino 316:324 arg1 protein 331:337 a 332-amino acid protein 314:337 a 332-amino acid protein 314:337 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 47 theme 332-amino 316:324 arg1 A 347:347 torsin A 340:347 torsin A 340:347 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 47 theme 332-amino 316:324 arg1 gene 301:304 The DYT1 gene 292:304 The DYT1 gene 292:304 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 6 48 theme exclusive 1166:1174 arg1 structures 1155:1164 large spheroid intracellular structures 1126:1164 large spheroid intracellular structures exclusive of BiP immunoreactivity 1126:1198 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 2 49 theme torsin 340:345 arg1 protein 331:337 a 332-amino acid protein 314:337 a 332-amino acid protein 314:337 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 49 theme torsin 340:345 arg1 A 347:347 torsin A 340:347 torsin A 340:347 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 1 50 theme hyperkinetic 187:198 arg1 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 50 theme hyperkinetic 187:198 arg1 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 6 51 theme intracellular 1141:1153 arg1 structures 1155:1164 large spheroid intracellular structures 1126:1164 large spheroid intracellular structures exclusive of BiP immunoreactivity 1126:1198 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 10 52 theme protein 1799:1805 arg1 phenotype 1782:1790 the disease-inducing phenotype 1761:1790 the disease-inducing phenotype of the protein 1761:1805 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 10 52 theme protein 1799:1805 arg1 distribution 1725:1736 the altered subcellular distribution 1701:1736 the altered subcellular distribution of DeltaE-torsin A 1701:1755 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 3 53 theme glutamic 531:538 arg1 residues 545:552 glutamic acid residues 531:552 glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A) 531:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 5 54 theme torsin 801:806 arg1 A 808:808 torsin A 801:808 torsin A 801:808 When transfected into mammalian cells, both torsin A and DeltaE-torsin A were found to behave as lumenally oriented glycoproteins. 10871631 6 55 theme large 1126:1130 arg1 structures 1155:1164 large spheroid intracellular structures 1126:1164 large spheroid intracellular structures exclusive of BiP immunoreactivity 1126:1198 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 9 56 dep folded 1611:1616 arg1 lumenal 1619:1625 lumenal 1619:1625 lumenal 1619:1625 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 3 57 theme pair 523:526 arg1 one 514:516 one 514:516 one 514:516 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 3 57 theme pair 523:526 arg1 pair 523:526 a pair 521:526 a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A) 521:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 1 58 from deletion 265:272 arg1 gene 286:289 the DYT1 gene 277:289 the DYT1 gene 277:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 5 59 theme mammalian 779:787 arg1 cells 789:793 mammalian cells 779:793 mammalian cells 779:793 When transfected into mammalian cells, both torsin A and DeltaE-torsin A were found to behave as lumenally oriented glycoproteins. 10871631 8 60 theme sucrose 1472:1478 arg1 gradients 1480:1488 sucrose gradients 1472:1488 sucrose gradients 1472:1488 However, both torsin A and DeltaE-torsin A were readily solubilized by nonionic detergents, were similarly accessible to proteases, and displayed equivalent migration patterns on sucrose gradients. 10871631 3 61 theme residues 545:552 arg1 pair 523:526 a pair 521:526 a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A) 521:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 1 62 theme torsion 145:151 arg1 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 62 theme torsion 145:151 arg1 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 0 63 theme lumenal 62:68 arg1 glycoproteins 70:82 lumenal glycoproteins 62:82 lumenal glycoproteins that exhibit distinct subcellular localizations 62:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 63 theme lumenal 62:68 arg1 A 7:7 Torsin A 0:7 Torsin A 0:7 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 63 theme lumenal 62:68 arg1 forms 52:56 its torsion dystonia-associated mutant forms 13:56 its torsion dystonia-associated mutant forms 13:56 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 3 64 theme C 572:572 arg1 terminus 574:581 the C terminus 568:581 the C terminus of torsin A (DeltaE-torsin A) 568:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 6 65 theme diffuse 953:959 arg1 network 961:967 a diffuse network 951:967 a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP 951:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 6 66 theme BiP 1179:1181 arg1 immunoreactivity 1183:1198 BiP immunoreactivity 1179:1198 BiP immunoreactivity 1179:1198 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 1 67 theme autosomal 168:176 arg1 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 67 theme autosomal 168:176 arg1 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 8 68 theme DeltaE-torsin 1320:1332 arg1 A 1334:1334 DeltaE-torsin A 1320:1334 DeltaE-torsin A 1320:1334 However, both torsin A and DeltaE-torsin A were readily solubilized by nonionic detergents, were similarly accessible to proteases, and displayed equivalent migration patterns on sucrose gradients. 10871631 0 69 theme distinct 97:104 arg1 localizations 118:130 distinct subcellular localizations 97:130 distinct subcellular localizations 97:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 10 70 theme subcellular 1713:1723 arg1 distribution 1725:1736 the altered subcellular distribution 1701:1736 the altered subcellular distribution of DeltaE-torsin A 1701:1755 The potential relationship between the altered subcellular distribution of DeltaE-torsin A and the disease-inducing phenotype of the protein is discussed. 10871631 9 71 theme dystonia-associated 1560:1578 arg1 forms 1580:1584 torsion dystonia-associated forms 1552:1584 torsion dystonia-associated forms of torsin A 1552:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 71 theme dystonia-associated 1560:1578 arg1 proteins 1627:1634 properly folded, lumenal proteins 1602:1634 properly folded, lumenal proteins of similar oligomeric states 1602:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 71 theme dystonia-associated 1560:1578 arg1 type 1543:1546 the wild type 1534:1546 the wild type 1534:1546 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 6 72 theme endoplasmic 1047:1057 arg1 reticulum 1059:1067 the endoplasmic reticulum 1043:1067 the endoplasmic reticulum resident protein BiP 1043:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 5 73 gly glycoproteins 873:885 arg1 glycoproteins 873:885 lumenally oriented glycoproteins 854:885 lumenally oriented glycoproteins 854:885 When transfected into mammalian cells, both torsin A and DeltaE-torsin A were found to behave as lumenally oriented glycoproteins. 10871631 2 74 theme significant 369:379 arg1 homology 381:388 low but significant homology 361:388 low but significant homology to the Hsp100/Clp family of ATPase chaperones 361:434 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 75 contain bears 355:359 arg2 homology 381:388 low but significant homology 361:388 low but significant homology to the Hsp100/Clp family of ATPase chaperones 361:434 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 75 contain bears 355:359 arg1 protein 331:337 a 332-amino acid protein 314:337 a 332-amino acid protein 314:337 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 75 contain bears 355:359 arg1 A 347:347 torsin A 340:347 torsin A 340:347 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 75 contain bears 355:359 arg1 gene 301:304 The DYT1 gene 292:304 The DYT1 gene 292:304 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 76 theme chaperones 425:434 arg1 family 408:413 the Hsp100/Clp family 393:413 the Hsp100/Clp family of ATPase chaperones 393:434 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 4 77 theme subcellular 664:674 arg1 distribution 676:687 subcellular distribution 664:687 subcellular distribution 664:687 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 2 78 theme low 361:363 arg1 homology 381:388 low but significant homology 361:388 low but significant homology to the Hsp100/Clp family of ATPase chaperones 361:434 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 4 79 theme DeltaE-torsin 732:744 arg1 protein 748:754 DeltaE-torsin A protein 732:754 DeltaE-torsin A protein 732:754 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 9 80 theme A 1596:1596 arg1 forms 1580:1584 torsion dystonia-associated forms 1552:1584 torsion dystonia-associated forms of torsin A 1552:1596 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 80 theme A 1596:1596 arg1 proteins 1627:1634 properly folded, lumenal proteins 1602:1634 properly folded, lumenal proteins of similar oligomeric states 1602:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 9 80 theme A 1596:1596 arg1 type 1543:1546 the wild type 1534:1546 the wild type 1534:1546 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 6 81 theme significant 1007:1017 arg1 co-immunoreactivity 1019:1037 significant co-immunoreactivity 1007:1037 significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP 1007:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 0 82 gly glycoproteins 70:82 arg1 glycoproteins 70:82 lumenal glycoproteins 62:82 lumenal glycoproteins that exhibit distinct subcellular localizations 62:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 82 gly glycoproteins 70:82 arg1 A 7:7 Torsin A 0:7 Torsin A 0:7 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 82 gly glycoproteins 70:82 arg1 forms 52:56 its torsion dystonia-associated mutant forms 13:56 its torsion dystonia-associated mutant forms 13:56 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 3 83 theme DeltaE-torsin 596:608 arg1 A 593:593 torsin A 586:593 torsin A (DeltaE-torsin A) 586:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 3 83 theme DeltaE-torsin 596:608 arg1 A 610:610 DeltaE-torsin A 596:610 DeltaE-torsin A 596:610 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 2 84 theme Hsp100/Clp 397:406 arg1 family 408:413 the Hsp100/Clp family 393:413 the Hsp100/Clp family of ATPase chaperones 393:434 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 0 85 theme dystonia-associated 25:43 arg1 glycoproteins 70:82 lumenal glycoproteins 62:82 lumenal glycoproteins that exhibit distinct subcellular localizations 62:130 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 85 theme dystonia-associated 25:43 arg1 A 7:7 Torsin A 0:7 Torsin A 0:7 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 0 85 theme dystonia-associated 25:43 arg1 forms 52:56 its torsion dystonia-associated mutant forms 13:56 its torsion dystonia-associated mutant forms 13:56 Torsin A and its torsion dystonia-associated mutant forms are lumenal glycoproteins that exhibit distinct subcellular localizations. 10871631 6 86 theme membranes 986:994 arg1 network 961:967 a diffuse network 951:967 a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP 951:1088 Immunofluorescence studies revealed that torsin A localized to a diffuse network of intracellular membranes displaying significant co-immunoreactivity for the endoplasmic reticulum resident protein BiP, whereas DeltaE-torsin A resided in large spheroid intracellular structures exclusive of BiP immunoreactivity. 10871631 3 87 theme A 593:593 arg1 terminus 574:581 the C terminus 568:581 the C terminus of torsin A (DeltaE-torsin A) 568:611 The deletion in DYT1 associated with torsion dystonia results in the loss of one of a pair of glutamic acid residues residing near the C terminus of torsin A (DeltaE-torsin A). 10871631 4 88 theme A 746:746 arg1 protein 748:754 DeltaE-torsin A protein 732:754 DeltaE-torsin A protein 732:754 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 1 89 attach linked 241:246 arg2 dystonia 153:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia 133:160 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 89 attach linked 241:246 arg1 deletion 265:272 a 3-base pair deletion 251:272 a 3-base pair deletion in the DYT1 gene 251:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 1 89 attach linked 241:246 arg2 disorder 209:216 an autosomal dominant hyperkinetic movement disorder 165:216 an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene 165:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 10871631 4 90 theme torsin 720:725 arg1 A 727:727 the torsin A 716:727 the torsin A 716:727 At present, little is known about the expression, subcellular distribution, and/or function of either the torsin A or DeltaE-torsin A protein. 10871631 2 91 theme acid 326:329 arg1 protein 331:337 a 332-amino acid protein 314:337 a 332-amino acid protein 314:337 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 91 theme acid 326:329 arg1 A 347:347 torsin A 340:347 torsin A 340:347 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 2 91 theme acid 326:329 arg1 gene 301:304 The DYT1 gene 292:304 The DYT1 gene 292:304 The DYT1 gene encodes a 332-amino acid protein, torsin A, that bears low but significant homology to the Hsp100/Clp family of ATPase chaperones. 10871631 9 92 theme similar 1639:1645 arg1 states 1658:1663 similar oligomeric states 1639:1663 similar oligomeric states 1639:1663 Collectively, these data support that both the wild type and torsion dystonia-associated forms of torsin A are properly folded, lumenal proteins of similar oligomeric states. 10871631 1 93 theme pair 260:263 arg1 deletion 265:272 a 3-base pair deletion 251:272 a 3-base pair deletion in the DYT1 gene 251:289 Early-onset torsion dystonia is an autosomal dominant hyperkinetic movement disorder that has recently been linked to a 3-base pair deletion in the DYT1 gene. 9539702 0 0 theme adhesion 76:83 arg1 molecule-1 85:94 intercellular adhesion molecule-1 62:94 intercellular adhesion molecule-1 62:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 7 1 theme described 1322:1330 arg1 domains 1338:1344 previously described I set domains 1311:1344 previously described I set domains 1311:1344 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 1 theme described 1322:1330 arg1 ICAM-1 1300:1305 ICAM-1 1300:1305 ICAM-1 1300:1305 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 4 2 from tip 711:713 arg1 BC 688:689 the flexible BC 675:689 the flexible BC 675:689 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 4 2 from tip 711:713 arg1 loops 698:702 FG loops 695:702 FG loops 695:702 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 1 3 theme intercellular 146:158 arg1 molecule-1 169:178 intercellular adhesion molecule-1 146:178 intercellular adhesion molecule-1 (ICAM-1, CD54) 146:193 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 6 4 from residues 1107:1114 arg1 domain 1119:1124 domain 1 1119:1126 domain 1 1119:1126 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 0 5 theme intercellular 62:74 arg1 molecule-1 85:94 intercellular adhesion molecule-1 62:94 intercellular adhesion molecule-1 62:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 2 6 from ICAM-1 427:432 arg1 surface 446:452 the cell surface 437:452 the cell surface 437:452 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 4 7 theme domain 760:765 arg1 half 752:755 the upper half 742:755 the upper half of domain 1 742:767 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 1 8 theme adhesion 160:167 arg1 molecule-1 169:178 intercellular adhesion molecule-1 146:178 intercellular adhesion molecule-1 (ICAM-1, CD54) 146:193 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 7 9 dep proposed 1270:1277 arg1 belong 1437:1442 belong 1437:1442 belong to the I2 set 1437:1456 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 9 dep proposed 1270:1277 arg1 belong 1346:1351 belong 1346:1351 belong to the I1 set 1346:1365 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 2 10 theme hydrophobic 341:351 arg1 interface 353:361 a hydrophobic interface 339:361 a hydrophobic interface on the BED sheet of domain 1 339:390 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 6 11 from presence 944:951 arg1 bonds 1011:1015 conserved hydrogen bonds 992:1015 conserved hydrogen bonds at domain junctions 992:1035 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 11 from presence 944:951 arg1 domains 975:981 both domains 1 and 2 970:989 both domains 1 and 2 970:989 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 3 12 theme bivalent 522:529 arg1 binding 531:537 bivalent binding 522:537 bivalent binding in the dimer 522:550 The residues that bind to the integrin LFA-1 are well oriented for bivalent binding in the dimer, with the critical Glu-34 residues pointing away from each other on the periphery. 9539702 0 13 theme molecule-1 85:94 arg1 domains 51:57 the N-terminal two domains 32:57 the N-terminal two domains of intercellular adhesion molecule-1 32:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 0 13 theme molecule-1 85:94 arg1 molecule-1 85:94 intercellular adhesion molecule-1 62:94 intercellular adhesion molecule-1 62:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 4 14 theme FG 695:696 arg1 loops 698:702 FG loops 695:702 FG loops 695:702 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 2 15 theme independent 257:267 arg1 molecules 269:277 two independent molecules 253:277 two independent molecules 253:277 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 6 16 from junctions 1027:1035 arg1 bonds 1011:1015 conserved hydrogen bonds 992:1015 conserved hydrogen bonds at domain junctions 992:1035 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 16 from junctions 1027:1035 arg1 domains 975:981 both domains 1 and 2 970:989 both domains 1 and 2 970:989 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 1 17 theme 190-residue 122:132 arg1 fragment 134:141 a 190-residue fragment 120:141 a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) 120:193 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 4 18 theme upper 746:750 arg1 half 752:755 the upper half 742:755 the upper half of domain 1 742:767 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 5 19 theme important 844:852 arg1 residue 836:842 a residue 834:842 a residue important for binding to Plasmodium falciparum-infected erythrocytes 834:911 By contrast, a residue important for binding to Plasmodium falciparum-infected erythrocytes is in the dimer interface. 9539702 1 20 theme molecule-1 169:178 arg1 fragment 134:141 a 190-residue fragment 120:141 a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) 120:193 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 2 21 from surface 446:452 arg1 dimerization 411:422 dimerization 411:422 dimerization of ICAM-1 on the cell surface 411:452 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 2 22 from interface 353:361 arg1 sheet 374:378 the BED sheet 366:378 the BED sheet of domain 1 366:390 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 6 23 theme domain 1020:1025 arg1 junctions 1027:1035 domain junctions 1020:1035 domain junctions 1020:1035 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 24 theme strands 959:965 arg1 presence 944:951 The presence 940:951 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, 940:1036 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 0 25 theme dimeric 2:8 arg1 structure 18:26 A dimeric crystal structure 0:26 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1 0:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 3 26 theme integrin 485:492 arg1 LFA-1 494:498 the integrin LFA-1 481:498 the integrin LFA-1 481:498 The residues that bind to the integrin LFA-1 are well oriented for bivalent binding in the dimer, with the critical Glu-34 residues pointing away from each other on the periphery. 9539702 6 27 dep domains 975:981 arg1 2 989:989 2 989:989 2 989:989 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 27 dep domains 975:981 arg1 1 983:983 1 983:983 1 983:983 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 4 28 theme flexible 679:686 arg1 BC 688:689 the flexible BC 675:689 the flexible BC 675:689 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 7 29 theme ICAM-1 1300:1305 arg1 domains 1338:1344 previously described I set domains 1311:1344 previously described I set domains 1311:1344 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 29 theme ICAM-1 1300:1305 arg1 domain 1288:1293 domain 1 1288:1295 domain 1 of ICAM-1 1288:1305 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 29 theme ICAM-1 1300:1305 arg1 ICAM-1 1300:1305 ICAM-1 1300:1305 ICAM-1 1300:1305 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 6 30 theme binding 1099:1105 arg1 residues 1107:1114 the key integrin binding residues 1082:1114 the key integrin binding residues in domain 1 1082:1126 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 7 31 theme IgSF 1254:1257 arg1 domains 1259:1265 IgSF domains 1254:1265 IgSF domains 1254:1265 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 2 32 theme BED 370:372 arg1 sheet 374:378 the BED sheet 366:378 the BED sheet of domain 1 366:390 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 6 33 theme hydrogen 1052:1059 arg1 networks 1066:1073 hydrogen bond networks 1052:1073 hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions 1052:1206 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 34 theme conserved 992:1000 arg1 bonds 1011:1015 conserved hydrogen bonds 992:1015 conserved hydrogen bonds at domain junctions 992:1035 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 34 theme conserved 992:1000 arg1 domains 975:981 both domains 1 and 2 970:989 both domains 1 and 2 970:989 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 35 theme hydrogen 1002:1009 arg1 bonds 1011:1015 conserved hydrogen bonds 992:1015 conserved hydrogen bonds at domain junctions 992:1035 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 35 theme hydrogen 1002:1009 arg1 domains 975:981 both domains 1 and 2 970:989 both domains 1 and 2 970:989 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 7 36 theme ICAM-2 1391:1396 arg1 molecule-1 1426:1435 vascular cell adhesion molecule-1 1403:1435 vascular cell adhesion molecule-1 1403:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 36 theme ICAM-2 1391:1396 arg1 ICAM-2 1391:1396 ICAM-2 1391:1396 ICAM-2 1391:1396 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 36 theme ICAM-2 1391:1396 arg1 domain 1371:1376 domain 2 1371:1378 domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 1371:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 36 theme ICAM-2 1391:1396 arg1 ICAM-1 1383:1388 ICAM-1 1383:1388 ICAM-1 1383:1388 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 0 37 theme crystal 10:16 arg1 structure 18:26 A dimeric crystal structure 0:26 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1 0:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 7 38 theme molecule-1 1426:1435 arg1 molecule-1 1426:1435 vascular cell adhesion molecule-1 1403:1435 vascular cell adhesion molecule-1 1403:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 38 theme molecule-1 1426:1435 arg1 ICAM-2 1391:1396 ICAM-2 1391:1396 ICAM-2 1391:1396 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 38 theme molecule-1 1426:1435 arg1 domain 1371:1376 domain 2 1371:1378 domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 1371:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 38 theme molecule-1 1426:1435 arg1 ICAM-1 1383:1388 ICAM-1 1383:1388 ICAM-1 1383:1388 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 6 39 theme adhesive 1186:1193 arg1 interactions 1195:1206 adhesive interactions 1186:1206 adhesive interactions 1186:1206 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 7 40 theme I1 1360:1361 arg1 set 1363:1365 the I1 set 1356:1365 the I1 set 1356:1365 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 1 41 theme 3.0-A 101:105 arg1 structure 107:115 The 3.0-A structure 97:115 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) 97:193 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 7 42 theme I2 1451:1452 arg1 set 1454:1456 the I2 set 1447:1456 the I2 set 1447:1456 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 5 43 theme falciparum-infected 880:898 arg1 erythrocytes 900:911 Plasmodium falciparum-infected erythrocytes 869:911 Plasmodium falciparum-infected erythrocytes 869:911 By contrast, a residue important for binding to Plasmodium falciparum-infected erythrocytes is in the dimer interface. 9539702 6 44 theme integrin 1090:1097 arg1 residues 1107:1114 the key integrin binding residues 1082:1114 the key integrin binding residues in domain 1 1082:1126 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 2 45 theme domain 383:388 arg1 sheet 374:378 the BED sheet 366:378 the BED sheet of domain 1 366:390 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 7 46 theme cell 1412:1415 arg1 molecule-1 1426:1435 vascular cell adhesion molecule-1 1403:1435 vascular cell adhesion molecule-1 1403:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 2 47 theme cell 441:444 arg1 surface 446:452 the cell surface 437:452 the cell surface 437:452 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 6 48 theme key 1086:1088 arg1 residues 1107:1114 the key integrin binding residues 1082:1114 the key integrin binding residues in domain 1 1082:1126 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 7 49 theme intermediate 1230:1241 arg1 set 1247:1249 the intermediate (I) set 1226:1249 the intermediate (I) set of IgSF domains 1226:1265 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 49 theme intermediate 1230:1241 arg1 domains 1259:1265 IgSF domains 1254:1265 IgSF domains 1254:1265 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 49 theme intermediate 1230:1241 arg1 I 1244:1244 I 1244:1244 I 1244:1244 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 50 theme vascular 1403:1410 arg1 molecule-1 1426:1435 vascular cell adhesion molecule-1 1403:1435 vascular cell adhesion molecule-1 1403:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 51 theme adhesion 1417:1424 arg1 molecule-1 1426:1435 vascular cell adhesion molecule-1 1403:1435 vascular cell adhesion molecule-1 1403:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 6 52 theme tensile 1164:1170 arg1 forces 1172:1177 tensile forces 1164:1177 tensile forces during adhesive interactions 1164:1206 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 1 53 dep molecule-1 169:178 arg1 ICAM-1 181:186 ICAM-1 181:186 ICAM-1 181:186 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 1 53 dep molecule-1 169:178 arg1 CD54 189:192 CD54 189:192 CD54 189:192 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 2 54 with agreement 396:404 arg1 dimerization 411:422 dimerization 411:422 dimerization of ICAM-1 on the cell surface 411:452 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 0 55 theme N-terminal 36:45 arg1 domains 51:57 the N-terminal two domains 32:57 the N-terminal two domains of intercellular adhesion molecule-1 32:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 0 55 theme N-terminal 36:45 arg1 molecule-1 85:94 intercellular adhesion molecule-1 62:94 intercellular adhesion molecule-1 62:94 A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. 9539702 5 56 theme dimer 923:927 arg1 interface 929:937 the dimer interface 919:937 the dimer interface 919:937 By contrast, a residue important for binding to Plasmodium falciparum-infected erythrocytes is in the dimer interface. 9539702 7 57 theme set 1247:1249 arg1 subdivision 1211:1221 A subdivision 1209:1221 A subdivision of the intermediate (I) set of IgSF domains 1209:1265 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 4 58 theme domain 718:723 arg1 tip 711:713 the tip 707:713 the tip of domain 1 707:725 Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. 9539702 3 59 theme Glu-34 571:576 arg1 residues 578:585 the critical Glu-34 residues 558:585 the critical Glu-34 residues pointing away from each other on the periphery 558:632 The residues that bind to the integrin LFA-1 are well oriented for bivalent binding in the dimer, with the critical Glu-34 residues pointing away from each other on the periphery. 9539702 7 60 theme domains 1259:1265 arg1 set 1247:1249 the intermediate (I) set 1226:1249 the intermediate (I) set of IgSF domains 1226:1265 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 60 theme domains 1259:1265 arg1 domains 1259:1265 IgSF domains 1254:1265 IgSF domains 1254:1265 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 60 theme domains 1259:1265 arg1 I 1244:1244 I 1244:1244 I 1244:1244 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 6 61 dep networks 1066:1073 arg1 make 1128:1131 make 1128:1131 make these domains suited to resist tensile forces during adhesive interactions 1128:1206 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 1 62 theme tandem 207:212 arg1 domains 236:242 two tandem Ig-superfamily (IgSF) domains 203:242 two tandem Ig-superfamily (IgSF) domains 203:242 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 7 63 theme I 1332:1332 arg1 domains 1338:1344 previously described I set domains 1311:1344 previously described I set domains 1311:1344 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 63 theme I 1332:1332 arg1 ICAM-1 1300:1305 ICAM-1 1300:1305 ICAM-1 1300:1305 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 6 64 theme bond 1061:1064 arg1 networks 1066:1073 hydrogen bond networks 1052:1073 hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions 1052:1206 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 2 65 theme symmetry-related 308:323 arg1 molecule 325:332 a symmetry-related molecule 306:332 a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1 306:390 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 3 66 from binding 531:537 arg1 dimer 546:550 the dimer 542:550 the dimer 542:550 The residues that bind to the integrin LFA-1 are well oriented for bivalent binding in the dimer, with the critical Glu-34 residues pointing away from each other on the periphery. 9539702 1 67 theme Ig-superfamily 214:227 arg1 domains 236:242 two tandem Ig-superfamily (IgSF) domains 203:242 two tandem Ig-superfamily (IgSF) domains 203:242 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 7 68 theme set 1334:1336 arg1 domains 1338:1344 previously described I set domains 1311:1344 previously described I set domains 1311:1344 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 68 theme set 1334:1336 arg1 ICAM-1 1300:1305 ICAM-1 1300:1305 ICAM-1 1300:1305 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 5 69 theme Plasmodium 869:878 arg1 erythrocytes 900:911 Plasmodium falciparum-infected erythrocytes 869:911 Plasmodium falciparum-infected erythrocytes 869:911 By contrast, a residue important for binding to Plasmodium falciparum-infected erythrocytes is in the dimer interface. 9539702 6 70 from strands 959:965 arg1 bonds 1011:1015 conserved hydrogen bonds 992:1015 conserved hydrogen bonds at domain junctions 992:1035 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 6 70 from strands 959:965 arg1 domains 975:981 both domains 1 and 2 970:989 both domains 1 and 2 970:989 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 1 71 theme fragment 134:141 arg1 structure 107:115 The 3.0-A structure 97:115 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) 97:193 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 9539702 7 72 theme ICAM-1 1383:1388 arg1 molecule-1 1426:1435 vascular cell adhesion molecule-1 1403:1435 vascular cell adhesion molecule-1 1403:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 72 theme ICAM-1 1383:1388 arg1 ICAM-2 1391:1396 ICAM-2 1391:1396 ICAM-2 1391:1396 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 72 theme ICAM-1 1383:1388 arg1 domain 1371:1376 domain 2 1371:1378 domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 1371:1435 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 7 72 theme ICAM-1 1383:1388 arg1 ICAM-1 1383:1388 ICAM-1 1383:1388 ICAM-1 1383:1388 A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set. 9539702 2 73 from dimerization 411:422 arg1 surface 446:452 the cell surface 437:452 the cell surface 437:452 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 3 74 theme critical 562:569 arg1 residues 578:585 the critical Glu-34 residues 558:585 the critical Glu-34 residues pointing away from each other on the periphery 558:632 The residues that bind to the integrin LFA-1 are well oriented for bivalent binding in the dimer, with the critical Glu-34 residues pointing away from each other on the periphery. 9539702 6 75 from domains 975:981 arg1 presence 944:951 The presence 940:951 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, 940:1036 The presence of A' strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. 9539702 2 76 theme ICAM-1 427:432 arg1 dimerization 411:422 dimerization 411:422 dimerization of ICAM-1 on the cell surface 411:452 Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. 9539702 1 77 theme IgSF 230:233 arg1 domains 236:242 two tandem Ig-superfamily (IgSF) domains 203:242 two tandem Ig-superfamily (IgSF) domains 203:242 The 3.0-A structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. 12526797 3 0 theme allosteric 641:650 arg1 signals 652:658 allosteric signals 641:658 allosteric signals 641:658 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 4 1 theme disulfide 819:827 arg1 bonds 829:833 introduced disulfide bonds 808:833 introduced disulfide bonds 808:833 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 4 2 theme 6-alpha 853:859 arg1 loop 863:866 the beta 6-alpha 7 loop 844:866 the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations 844:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 4 3 theme introduced 808:817 arg1 bonds 829:833 introduced disulfide bonds 808:833 introduced disulfide bonds 808:833 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 1 4 theme ligand 205:210 arg1 ICAM-1 212:217 ligand ICAM-1 205:217 the Ig superfamily ligand ICAM-1 186:217 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 1 5 theme integrin-IgSF 292:304 arg1 interface 306:314 an integrin-IgSF interface 289:314 an integrin-IgSF interface 289:314 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 4 6 theme C-terminal 778:787 arg1 helix 797:801 the C-terminal alpha 7 helix 774:801 the C-terminal alpha 7 helix 774:801 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 4 7 with down 766:769 arg1 bonds 829:833 introduced disulfide bonds 808:833 introduced disulfide bonds 808:833 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 1 8 theme interface 306:314 arg1 example 278:284 the first example 268:284 the first example of an integrin-IgSF interface 268:314 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 1 8 theme interface 306:314 arg1 conformation 251:262 the open ligand binding conformation 227:262 the open ligand binding conformation 227:262 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 0 9 with complex 43:49 arg1 ICAM-1 56:61 ICAM-1 56:61 ICAM-1 56:61 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 4 10 theme open 932:935 arg1 conformations 937:949 the closed, intermediate, and open conformations 902:949 the closed, intermediate, and open conformations 902:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 4 11 from positions 889:897 arg1 conformations 937:949 the closed, intermediate, and open conformations 902:949 the closed, intermediate, and open conformations 902:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 4 12 theme closed 906:911 arg1 conformations 937:949 the closed, intermediate, and open conformations 902:949 the closed, intermediate, and open conformations 902:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 2 13 theme ICAM-1 366:371 arg1 Glu-34 356:361 Glu-34 356:361 Glu-34 of ICAM-1 356:371 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 1 14 theme I 141:141 arg1 beta 170:173 integrin alpha L beta 2 153:175 integrin alpha L beta 2 153:175 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 1 14 theme I 141:141 arg1 domain 143:148 the I domain 137:148 the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 137:217 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 0 15 theme shape-shifting 72:85 arg1 pathway 87:93 a shape-shifting pathway 70:93 a shape-shifting pathway for integrin regulation 70:117 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 2 16 theme residue 407:413 arg1 swing 389:393 a dramatic swing 378:393 a dramatic swing of I domain residue Glu-241 378:421 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 3 17 theme unliganded 468:477 arg1 structures 479:488 Liganded and unliganded structures 455:488 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains 455:546 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 1 18 theme domain 143:148 arg1 structure 124:132 The structure 120:132 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 120:217 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 4 19 theme progressive 959:969 arg1 increase 971:978 a progressive increase 957:978 a progressive increase in affinity 957:990 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 4 20 theme intermediate 914:925 arg1 conformations 937:949 the closed, intermediate, and open conformations 902:949 the closed, intermediate, and open conformations 902:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 2 21 theme dramatic 380:387 arg1 swing 389:393 a dramatic swing 378:393 a dramatic swing of I domain residue Glu-241 378:421 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 4 22 from down 766:769 arg1 helix 797:801 the C-terminal alpha 7 helix 774:801 the C-terminal alpha 7 helix 774:801 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 3 23 theme intermediate-affinity 509:529 arg1 domains 540:546 both high- and intermediate-affinity mutant I domains 494:546 both high- and intermediate-affinity mutant I domains 494:546 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 1 24 theme integrin 153:160 arg1 beta 170:173 integrin alpha L beta 2 153:175 integrin alpha L beta 2 153:175 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 0 25 theme alpha 18:22 arg1 domain 28:33 the alpha L I domain 14:33 the alpha L I domain 14:33 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 1 26 theme open 231:234 arg1 conformation 251:262 the open ligand binding conformation 227:262 the open ligand binding conformation 227:262 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 0 27 theme integrin 99:106 arg1 regulation 108:117 integrin regulation 99:117 integrin regulation 99:117 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 1 28 theme alpha 162:166 arg1 beta 170:173 integrin alpha L beta 2 153:175 integrin alpha L beta 2 153:175 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 1 29 theme ligand 236:241 arg1 conformation 251:262 the open ligand binding conformation 227:262 the open ligand binding conformation 227:262 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 3 30 theme high- 499:503 arg1 domains 540:546 both high- and intermediate-affinity mutant I domains 494:546 both high- and intermediate-affinity mutant I domains 494:546 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 1 31 theme L 168:168 arg1 beta 170:173 integrin alpha L beta 2 153:175 integrin alpha L beta 2 153:175 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 0 32 theme I 26:26 arg1 domain 28:33 the alpha L I domain 14:33 the alpha L I domain 14:33 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 1 33 theme binding 243:249 arg1 conformation 251:262 the open ligand binding conformation 227:262 the open ligand binding conformation 227:262 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 4 34 theme different 879:887 arg1 positions 889:897 three different positions 873:897 three different positions in the closed, intermediate, and open conformations 873:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 2 35 theme domain 323:328 arg1 Mg2+ 330:333 The I domain Mg2+ 317:333 The I domain Mg2+ 317:333 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 1 36 theme beta 170:173 arg1 beta 170:173 integrin alpha L beta 2 153:175 integrin alpha L beta 2 153:175 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 1 36 theme beta 170:173 arg1 domain 143:148 the I domain 137:148 the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 137:217 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 0 37 theme L 24:24 arg1 domain 28:33 the alpha L I domain 14:33 the alpha L I domain 14:33 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 2 38 theme domain 400:405 arg1 residue 407:413 I domain residue Glu-241 398:421 I domain residue Glu-241 398:421 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 2 39 theme salt 442:445 arg1 bridge 447:452 a critical salt bridge 431:452 a critical salt bridge 431:452 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 2 40 theme I 398:398 arg1 residue 407:413 I domain residue Glu-241 398:421 I domain residue Glu-241 398:421 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 4 41 theme beta 848:851 arg1 loop 863:866 the beta 6-alpha 7 loop 844:866 the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations 844:949 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 3 42 theme conformational 586:599 arg1 change 601:606 conformational change 586:606 conformational change 586:606 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 3 43 theme mutant 531:536 arg1 domains 540:546 both high- and intermediate-affinity mutant I domains 494:546 both high- and intermediate-affinity mutant I domains 494:546 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 3 44 theme intermediate 699:710 arg1 conformations 720:732 intermediate or open conformations 699:732 intermediate or open conformations without ligand binding 699:755 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 0 45 theme domain 28:33 arg1 Structures 0:9 Structures 0:9 Structures of the alpha L I domain 0:33 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 0 45 theme domain 28:33 arg1 complex 43:49 its complex 39:49 its complex with ICAM-1 39:61 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 4 46 theme alpha 789:793 arg1 helix 797:801 the C-terminal alpha 7 helix 774:801 the C-terminal alpha 7 helix 774:801 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 3 47 theme L 621:621 arg1 domain 625:630 the alpha L I domain 611:630 the alpha L I domain 611:630 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 3 48 theme I 538:538 arg1 domains 540:546 both high- and intermediate-affinity mutant I domains 494:546 both high- and intermediate-affinity mutant I domains 494:546 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 1 49 theme first 272:276 arg1 example 278:284 the first example 268:284 the first example of an integrin-IgSF interface 268:314 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 4 50 from increase 971:978 arg1 affinity 983:990 affinity 983:990 affinity 983:990 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 3 51 theme I 623:623 arg1 domain 625:630 the alpha L I domain 611:630 the alpha L I domain 611:630 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 0 52 with Structures 0:9 arg1 ICAM-1 56:61 ICAM-1 56:61 ICAM-1 56:61 Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. 12526797 3 53 theme closed 676:681 arg1 conformation 683:694 the closed conformation 672:694 the closed conformation 672:694 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 3 54 theme open 715:718 arg1 conformations 720:732 intermediate or open conformations 699:732 intermediate or open conformations without ligand binding 699:755 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 1 55 dep superfamily 193:203 arg1 ICAM-1 212:217 ligand ICAM-1 205:217 the Ig superfamily ligand ICAM-1 186:217 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 12526797 2 56 theme critical 433:440 arg1 bridge 447:452 a critical salt bridge 431:452 a critical salt bridge 431:452 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 3 57 theme alpha 615:619 arg1 domain 625:630 the alpha L I domain 611:630 the alpha L I domain 611:630 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 4 58 theme Pulling 758:764 arg1 down 766:769 Pulling down 758:769 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds 758:833 Pulling down on the C-terminal alpha 7 helix with introduced disulfide bonds ratchets the beta 6-alpha 7 loop into three different positions in the closed, intermediate, and open conformations, with a progressive increase in affinity. 12526797 2 59 theme I 321:321 arg1 Mg2+ 330:333 The I domain Mg2+ 317:333 The I domain Mg2+ 317:333 The I domain Mg2+ directly coordinates Glu-34 of ICAM-1, and a dramatic swing of I domain residue Glu-241 enables a critical salt bridge. 12526797 3 60 theme ligand 742:747 arg1 binding 749:755 ligand binding 742:755 ligand binding 742:755 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 3 61 theme ligand 560:565 arg1 binding 567:573 ligand binding 560:573 ligand binding 560:573 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 3 62 theme Liganded 455:462 arg1 structures 479:488 Liganded and unliganded structures 455:488 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains 455:546 Liganded and unliganded structures for both high- and intermediate-affinity mutant I domains reveal that ligand binding can induce conformational change in the alpha L I domain and that allosteric signals can convert the closed conformation to intermediate or open conformations without ligand binding. 12526797 1 63 theme Ig 190:191 arg1 superfamily 193:203 the Ig superfamily ligand ICAM-1 186:217 the Ig superfamily ligand ICAM-1 186:217 The structure of the I domain of integrin alpha L beta 2 bound to the Ig superfamily ligand ICAM-1 reveals the open ligand binding conformation and the first example of an integrin-IgSF interface. 22333914 2 0 theme INducing 479:486 arg1 domain 495:500 GPCR-Autoproteolysis INducing (GAIN) domain 458:500 GPCR-Autoproteolysis INducing (GAIN) domain 458:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 4 1 theme cell-adhesion 845:857 arg1 GPCRs 859:863 all human cell-adhesion GPCRs 835:863 all human cell-adhesion GPCRs 835:863 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 5 2 theme overall 1073:1079 arg1 domain 1086:1091 the overall GAIN domain 1069:1091 the overall GAIN domain 1069:1091 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 1 3 theme conserved 239:247 arg1 sequence 265:272 a highly conserved autoproteolysis sequence 230:272 a highly conserved autoproteolysis sequence 230:272 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 4 4 theme disease 918:924 arg1 mutations 926:934 multiple human disease mutations 903:934 multiple human disease mutations 903:934 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 4 5 theme PKD 869:871 arg1 proteins 873:880 PKD proteins 869:880 PKD proteins 869:880 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 3 6 theme Crystal 503:509 arg1 structures 511:520 Crystal structures 503:520 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs 503:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 3 7 theme domains 530:536 arg1 structures 511:520 Crystal structures 503:520 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs 503:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 1 8 theme autoproteolysis 249:263 arg1 sequence 265:272 a highly conserved autoproteolysis sequence 230:272 a highly conserved autoproteolysis sequence 230:272 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 4 9 theme GAIN 723:726 arg1 domain 728:733 The GAIN domain 719:733 The GAIN domain 719:733 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 5 10 theme GAIN 1081:1084 arg1 domain 1086:1091 the overall GAIN domain 1069:1091 the overall GAIN domain 1069:1091 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 6 11 theme autoproteolytic 1256:1270 arg1 fold 1272:1275 a unique, evolutionarily ancient and widespread autoproteolytic fold 1208:1275 a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases 1208:1361 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 6 12 theme human 1348:1352 arg1 diseases 1354:1361 multiple human diseases 1339:1361 multiple human diseases 1339:1361 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 3 13 from structures 511:520 arg1 GPCRs 579:583 two distantly related cell-adhesion GPCRs 543:583 two distantly related cell-adhesion GPCRs 543:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 1 14 theme cell-adhesion 153:165 arg1 GPCRs 167:171 cell-adhesion GPCRs 153:171 cell-adhesion GPCRs 153:171 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 4 15 theme mutations 926:934 arg1 locus 894:898 the locus 890:898 the locus of multiple human disease mutations 890:934 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 3 16 theme overall 698:704 arg1 domain 711:716 the overall GAIN domain 694:716 the overall GAIN domain 694:716 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 6 17 theme multiple 1339:1346 arg1 diseases 1354:1361 multiple human diseases 1339:1361 multiple human diseases 1339:1361 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 3 18 theme GAIN 525:528 arg1 domains 530:536 GAIN domains 525:536 GAIN domains from two distantly related cell-adhesion GPCRs 525:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 1 19 theme GPCRs 167:171 arg1 GPS 145:147 GPS 145:147 GPS 145:147 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 19 theme GPCRs 167:171 arg1 Site 139:142 The G protein-coupled receptor (GPCR) Proteolysis Site 89:142 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins 89:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 19 theme GPCRs 167:171 arg1 proteins 209:216 polycystic kidney disease (PKD) proteins 177:216 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 19 theme GPCRs 167:171 arg1 GPCRs 167:171 cell-adhesion GPCRs 153:171 cell-adhesion GPCRs 153:171 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 0 20 theme novel 2:6 arg1 domain 33:38 A novel evolutionarily conserved domain 0:38 A novel evolutionarily conserved domain of cell-adhesion GPCRs 0:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 0 20 theme novel 2:6 arg1 GPCRs 57:61 cell-adhesion GPCRs 43:61 cell-adhesion GPCRs 43:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 3 21 theme GAIN 706:709 arg1 domain 711:716 the overall GAIN domain 694:716 the overall GAIN domain 694:716 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 4 22 theme human 839:843 arg1 GPCRs 859:863 all human cell-adhesion GPCRs 835:863 all human cell-adhesion GPCRs 835:863 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 3 23 theme related 557:563 arg1 GPCRs 579:583 two distantly related cell-adhesion GPCRs 543:583 two distantly related cell-adhesion GPCRs 543:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 2 24 theme ∼320-residue 423:434 arg1 domain 436:441 a much larger ∼320-residue domain 409:441 a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain 409:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 5 25 theme bond 1160:1163 arg1 hydrolysis 1165:1174 peptide bond hydrolysis 1152:1174 peptide bond hydrolysis 1152:1174 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 1 26 theme protein-coupled 95:109 arg1 GPCR 121:124 GPCR 121:124 GPCR 121:124 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 26 theme protein-coupled 95:109 arg1 receptor 111:118 The G protein-coupled receptor 89:118 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins 89:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 3 27 from GPCRs 579:583 arg1 domains 530:536 GAIN domains 525:536 GAIN domains from two distantly related cell-adhesion GPCRs 525:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 3 27 from GPCRs 579:583 arg1 structures 511:520 Crystal structures 503:520 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs 503:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 2 28 theme GPS 368:370 arg1 motif 372:376 unexpectedly the ∼40-residue GPS motif 339:376 unexpectedly the ∼40-residue GPS motif 339:376 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 0 29 theme conserved 23:31 arg1 domain 33:38 A novel evolutionarily conserved domain 0:38 A novel evolutionarily conserved domain of cell-adhesion GPCRs 0:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 0 29 theme conserved 23:31 arg1 GPCRs 57:61 cell-adhesion GPCRs 43:61 cell-adhesion GPCRs 43:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 1 30 theme receptor 111:118 arg1 GPS 145:147 GPS 145:147 GPS 145:147 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 30 theme receptor 111:118 arg1 Site 139:142 The G protein-coupled receptor (GPCR) Proteolysis Site 89:142 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins 89:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 30 theme receptor 111:118 arg1 proteins 209:216 polycystic kidney disease (PKD) proteins 177:216 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 30 theme receptor 111:118 arg1 GPCRs 167:171 cell-adhesion GPCRs 153:171 cell-adhesion GPCRs 153:171 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 31 theme kidney 188:193 arg1 disease 195:201 polycystic kidney disease 177:201 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 31 theme kidney 188:193 arg1 PKD 204:206 PKD 204:206 PKD 204:206 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 3 32 theme cell-adhesion 565:577 arg1 GPCRs 579:583 two distantly related cell-adhesion GPCRs 543:583 two distantly related cell-adhesion GPCRs 543:583 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 6 33 dep unique 1210:1215 arg1 widespread 1245:1254 widespread 1245:1254 widespread 1245:1254 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 6 33 dep unique 1210:1215 arg1 ancient 1233:1239 ancient 1233:1239 ancient 1233:1239 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 1 34 theme disease 195:201 arg1 proteins 209:216 polycystic kidney disease (PKD) proteins 177:216 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 2 35 theme GPCR-Autoproteolysis 458:477 arg1 domain 495:500 GPCR-Autoproteolysis INducing (GAIN) domain 458:500 GPCR-Autoproteolysis INducing (GAIN) domain 458:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 1 36 theme catalytic 283:291 arg1 mechanism 293:301 its catalytic mechanism 279:301 its catalytic mechanism 279:301 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 4 37 theme multiple 903:910 arg1 mutations 926:934 multiple human disease mutations 903:934 multiple human disease mutations 903:934 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 4 38 theme only 799:802 arg1 domain 818:823 the only extracellular domain 795:823 the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins 795:880 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 1 39 theme polycystic 177:186 arg1 disease 195:201 polycystic kidney disease 177:201 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 39 theme polycystic 177:186 arg1 PKD 204:206 PKD 204:206 PKD 204:206 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 6 40 theme GPCR 1315:1318 arg1 signalling 1320:1329 GPCR signalling 1315:1329 GPCR signalling 1315:1329 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 2 41 theme larger 416:421 arg1 domain 436:441 a much larger ∼320-residue domain 409:441 a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain 409:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 2 42 theme integral 392:399 arg1 part 401:404 an integral part 389:404 an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain 389:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 0 43 theme GPCRs 57:61 arg1 domain 33:38 A novel evolutionarily conserved domain 0:38 A novel evolutionarily conserved domain of cell-adhesion GPCRs 0:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 0 43 theme GPCRs 57:61 arg1 GPCRs 57:61 cell-adhesion GPCRs 43:61 cell-adhesion GPCRs 43:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 3 44 theme GPS 630:632 arg1 motif 634:638 the GPS motif 626:638 the GPS motif 626:638 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 5 45 from environment 1117:1127 arg1 GPS 1136:1138 the GPS to catalyse peptide bond hydrolysis 1132:1174 the GPS to catalyse peptide bond hydrolysis 1132:1174 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 0 46 theme cell-adhesion 43:55 arg1 GPCRs 57:61 cell-adhesion GPCRs 43:61 cell-adhesion GPCRs 43:61 A novel evolutionarily conserved domain of cell-adhesion GPCRs mediates autoproteolysis. 22333914 6 47 theme unique 1210:1215 arg1 fold 1272:1275 a unique, evolutionarily ancient and widespread autoproteolytic fold 1208:1275 a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases 1208:1361 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 4 48 theme extracellular 804:816 arg1 domain 818:823 the only extracellular domain 795:823 the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins 795:880 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 22333914 5 49 theme peptide 1152:1158 arg1 hydrolysis 1165:1174 peptide bond hydrolysis 1152:1174 peptide bond hydrolysis 1152:1174 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 3 50 theme conserved 596:604 arg1 fold 612:615 a conserved novel fold 594:615 a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain 594:716 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 1 51 theme Proteolysis 127:137 arg1 GPS 145:147 GPS 145:147 GPS 145:147 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 51 theme Proteolysis 127:137 arg1 Site 139:142 The G protein-coupled receptor (GPCR) Proteolysis Site 89:142 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins 89:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 51 theme Proteolysis 127:137 arg1 proteins 209:216 polycystic kidney disease (PKD) proteins 177:216 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 51 theme Proteolysis 127:137 arg1 GPCRs 167:171 cell-adhesion GPCRs 153:171 cell-adhesion GPCRs 153:171 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 5 52 theme GAIN 955:958 arg1 domain 960:965 the GAIN domain 951:965 the GAIN domain 951:965 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 5 52 theme GAIN 955:958 arg1 necessary 975:983 necessary 975:983 necessary 975:983 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 6 53 theme GAIN 1187:1190 arg1 domain 1192:1197 the GAIN domain 1183:1197 the GAIN domain 1183:1197 Thus, the GAIN domain embodies a unique, evolutionarily ancient and widespread autoproteolytic fold whose function is likely relevant for GPCR signalling and for multiple human diseases. 22333914 1 54 theme proteins 209:216 arg1 GPS 145:147 GPS 145:147 GPS 145:147 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 54 theme proteins 209:216 arg1 Site 139:142 The G protein-coupled receptor (GPCR) Proteolysis Site 89:142 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins 89:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 54 theme proteins 209:216 arg1 proteins 209:216 polycystic kidney disease (PKD) proteins 177:216 polycystic kidney disease (PKD) proteins 177:216 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 1 54 theme proteins 209:216 arg1 GPCRs 167:171 cell-adhesion GPCRs 153:171 cell-adhesion GPCRs 153:171 The G protein-coupled receptor (GPCR) Proteolysis Site (GPS) of cell-adhesion GPCRs and polycystic kidney disease (PKD) proteins constitutes a highly conserved autoproteolysis sequence, but its catalytic mechanism remains unknown. 22333914 5 55 theme autoproteolytic 1035:1049 arg1 mechanism 1051:1059 an autoproteolytic mechanism 1032:1059 an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis 1032:1174 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 3 56 theme novel 606:610 arg1 fold 612:615 a conserved novel fold 594:615 a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain 594:716 Crystal structures of GAIN domains from two distantly related cell-adhesion GPCRs revealed a conserved novel fold in which the GPS motif forms five β-strands that are tightly integrated into the overall GAIN domain. 22333914 2 57 theme ∼40-residue 356:366 arg1 motif 372:376 unexpectedly the ∼40-residue GPS motif 339:376 unexpectedly the ∼40-residue GPS motif 339:376 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 2 58 theme GAIN 489:492 arg1 domain 495:500 GPCR-Autoproteolysis INducing (GAIN) domain 458:500 GPCR-Autoproteolysis INducing (GAIN) domain 458:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 5 59 theme chemical 1108:1115 arg1 environment 1117:1127 the chemical environment 1104:1127 the chemical environment in the GPS to catalyse peptide bond hydrolysis 1104:1174 Functionally, the GAIN domain is both necessary and sufficient for autoproteolysis, suggesting an autoproteolytic mechanism whereby the overall GAIN domain fine-tunes the chemical environment in the GPS to catalyse peptide bond hydrolysis. 22333914 2 60 theme domain 436:441 arg1 part 401:404 an integral part 389:404 an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain 389:500 Here, we show that unexpectedly the ∼40-residue GPS motif represents an integral part of a much larger ∼320-residue domain that we termed GPCR-Autoproteolysis INducing (GAIN) domain. 22333914 4 61 theme human 912:916 arg1 mutations 926:934 multiple human disease mutations 903:934 multiple human disease mutations 903:934 The GAIN domain is evolutionarily conserved from tetrahymena to mammals, is the only extracellular domain shared by all human cell-adhesion GPCRs and PKD proteins, and is the locus of multiple human disease mutations. 28117447 6 0 theme binding 1056:1062 arg1 motif 1064:1068 the binding motif 1052:1068 the binding motif 1052:1068 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 7 1 theme specific 1177:1184 arg1 orientation 1205:1215 a specific integrin/pro-TGF-β orientation 1175:1215 a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit 1175:1368 Regions in and outside the highly interdigitated interface stabilize a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit. 28117447 1 2 theme extracellular 158:170 arg1 ligands 172:178 extracellular ligands 158:178 extracellular ligands 158:178 Integrins are adhesion receptors that transmit force across the plasma membrane between extracellular ligands and the actin cytoskeleton. 28117447 5 3 dep presence 704:711 arg1 the 700:702 the 700:702 the 700:702 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 7 4 theme integrin/pro-TGF-β 1186:1203 arg1 orientation 1205:1215 a specific integrin/pro-TGF-β orientation 1175:1215 a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit 1175:1368 Regions in and outside the highly interdigitated interface stabilize a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit. 28117447 7 5 theme integrin 1351:1358 arg1 β-subunit 1360:1368 the integrin β-subunit 1347:1368 the integrin β-subunit 1347:1368 Regions in and outside the highly interdigitated interface stabilize a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit. 28117447 7 6 theme tensile 1306:1312 arg1 force 1314:1318 actin-cytoskeleton-generated tensile force 1277:1318 actin-cytoskeleton-generated tensile force 1277:1318 Regions in and outside the highly interdigitated interface stabilize a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit. 28117447 6 7 theme macromolecular 1018:1031 arg1 components 1033:1042 macromolecular components 1018:1042 macromolecular components outside the binding motif 1018:1068 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 5 8 theme integrin 728:735 arg1 binding 737:743 integrin binding 728:743 integrin binding 728:743 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 5 9 theme prodomain 656:664 arg1 motif 683:687 the prodomain integrin-binding motif 652:687 the prodomain integrin-binding motif 652:687 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 2 10 theme TGF-β 345:349 arg1 factor 358:363 the TGF-β growth factor 341:363 the TGF-β growth factor 341:363 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 4 11 theme relevant 572:579 arg1 orientation 547:557 an orientation 544:557 an orientation biologically relevant for force-dependent release of TGF-β from latency 544:629 Here we show how integrin αVβ6 binds pro-TGF-β1 in an orientation biologically relevant for force-dependent release of TGF-β from latency. 28117447 5 12 theme binding 737:743 arg1 presence 704:711 presence 704:711 presence 704:711 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 5 12 theme binding 737:743 arg1 absence 717:723 absence 717:723 absence 717:723 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 6 13 theme integrin-bound 911:924 arg1 conformation 926:937 the integrin-bound conformation 907:937 the integrin-bound conformation 907:937 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 7 14 theme interdigitated 1140:1153 arg1 interface 1155:1163 the highly interdigitated interface 1129:1163 the highly interdigitated interface 1129:1163 Regions in and outside the highly interdigitated interface stabilize a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit. 28117447 3 15 theme macromolecular 415:428 arg1 ligands 430:436 macromolecular ligands 415:436 macromolecular ligands 415:436 However, we know little about how integrins bind macromolecular ligands in the extracellular matrix or transmit force to them. 28117447 7 16 theme actin-cytoskeleton-generated 1277:1304 arg1 force 1314:1318 actin-cytoskeleton-generated tensile force 1277:1318 actin-cytoskeleton-generated tensile force 1277:1318 Regions in and outside the highly interdigitated interface stabilize a specific integrin/pro-TGF-β orientation that defines the pathway through these macromolecules which actin-cytoskeleton-generated tensile force takes when applied through the integrin β-subunit. 28117447 8 17 theme TGF-β 1497:1501 arg1 prodomain 1503:1511 the TGF-β prodomain 1493:1511 the TGF-β prodomain 1493:1511 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 1 18 theme adhesion 84:91 arg1 Integrins 70:78 Integrins 70:78 Integrins 70:78 Integrins are adhesion receptors that transmit force across the plasma membrane between extracellular ligands and the actin cytoskeleton. 28117447 1 18 theme adhesion 84:91 arg1 receptors 93:101 adhesion receptors 84:101 adhesion receptors that transmit force across the plasma membrane between extracellular ligands and the actin cytoskeleton 84:205 Integrins are adhesion receptors that transmit force across the plasma membrane between extracellular ligands and the actin cytoskeleton. 28117447 1 19 theme actin 188:192 arg1 cytoskeleton 194:205 the actin cytoskeleton 184:205 the actin cytoskeleton 184:205 Integrins are adhesion receptors that transmit force across the plasma membrane between extracellular ligands and the actin cytoskeleton. 28117447 0 20 theme macromolecular 21:34 arg1 structure 36:44 macromolecular structure 21:44 macromolecular structure 21:44 Force interacts with macromolecular structure in activation of TGF-β. 28117447 8 21 theme β- 1529:1530 arg1 α-subunit 1540:1548 the β- and not α-subunit 1525:1548 the β- and not α-subunit of the integrin 1525:1564 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 8 22 theme not 1536:1538 arg1 α-subunit 1540:1548 the β- and not α-subunit 1525:1548 the β- and not α-subunit of the integrin 1525:1564 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 3 23 theme extracellular 445:457 arg1 matrix 459:464 the extracellular matrix 441:464 the extracellular matrix 441:464 However, we know little about how integrins bind macromolecular ligands in the extracellular matrix or transmit force to them. 28117447 8 24 theme force 1467:1471 arg1 application 1473:1483 force application 1467:1483 force application 1467:1483 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 4 25 from latency 623:629 arg1 release 601:607 force-dependent release 585:607 force-dependent release of TGF-β from latency 585:629 Here we show how integrin αVβ6 binds pro-TGF-β1 in an orientation biologically relevant for force-dependent release of TGF-β from latency. 28117447 8 26 theme activation 1402:1411 arg1 Simulations 1371:1381 Simulations 1371:1381 Simulations of force-dependent activation of TGF-β 1371:1420 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 2 27 theme factor-β1 249:257 arg1 pro-TGF-β1 270:279 pro-TGF-β1 270:279 pro-TGF-β1 270:279 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 2 27 theme factor-β1 249:257 arg1 precursor 259:267 the transforming growth factor-β1 precursor 225:267 the transforming growth factor-β1 precursor (pro-TGF-β1) 225:280 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 6 28 theme integrin 1084:1091 arg1 recognition 1093:1103 integrin recognition 1084:1103 integrin recognition 1084:1103 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 6 29 theme family 989:994 arg1 members 996:1002 earlier-evolving family members 972:1002 earlier-evolving family members 972:1002 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 2 30 theme growth 242:247 arg1 pro-TGF-β1 270:279 pro-TGF-β1 270:279 pro-TGF-β1 270:279 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 2 30 theme growth 242:247 arg1 precursor 259:267 the transforming growth factor-β1 precursor 225:267 the transforming growth factor-β1 precursor (pro-TGF-β1) 225:280 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 8 31 theme force-dependent 1386:1400 arg1 activation 1402:1411 force-dependent activation 1386:1411 force-dependent activation of TGF-β 1386:1420 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 6 32 theme earlier-evolving 972:987 arg1 members 996:1002 earlier-evolving family members 972:1002 earlier-evolving family members 972:1002 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 5 33 theme integrin-binding 666:681 arg1 motif 683:687 the prodomain integrin-binding motif 652:687 the prodomain integrin-binding motif 652:687 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 2 34 theme transforming 229:240 arg1 pro-TGF-β1 270:279 pro-TGF-β1 270:279 pro-TGF-β1 270:279 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 2 34 theme transforming 229:240 arg1 precursor 259:267 the transforming growth factor-β1 precursor 225:267 the transforming growth factor-β1 precursor (pro-TGF-β1) 225:280 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 8 35 theme TGF-β 1416:1420 arg1 activation 1402:1411 force-dependent activation 1386:1411 force-dependent activation of TGF-β 1386:1420 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 5 36 theme motif 683:687 arg1 conformation 636:647 The conformation 632:647 The conformation of the prodomain integrin-binding motif 632:687 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 6 37 theme Remodelled 855:864 arg1 residues 866:873 Remodelled residues 855:873 Remodelled residues outside the interface 855:895 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 6 38 theme similar 961:967 arg1 conformation 948:959 a conformation 946:959 a conformation similar to earlier-evolving family members 946:1002 Remodelled residues outside the interface stabilize the integrin-bound conformation, adopt a conformation similar to earlier-evolving family members, and show how macromolecular components outside the binding motif contribute to integrin recognition. 28117447 8 39 theme integrin 1557:1564 arg1 α-subunit 1540:1548 the β- and not α-subunit 1525:1548 the β- and not α-subunit of the integrin 1525:1564 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 8 40 theme evolutionary 1434:1445 arg1 specializations 1447:1461 evolutionary specializations 1434:1461 evolutionary specializations for force application 1434:1483 Simulations of force-dependent activation of TGF-β demonstrate evolutionary specializations for force application through the TGF-β prodomain and through the β- and not α-subunit of the integrin. 28117447 1 41 theme plasma 134:139 arg1 membrane 141:148 the plasma membrane 130:148 the plasma membrane 130:148 Integrins are adhesion receptors that transmit force across the plasma membrane between extracellular ligands and the actin cytoskeleton. 28117447 4 42 theme force-dependent 585:599 arg1 release 601:607 force-dependent release 585:607 force-dependent release of TGF-β from latency 585:629 Here we show how integrin αVβ6 binds pro-TGF-β1 in an orientation biologically relevant for force-dependent release of TGF-β from latency. 28117447 5 43 theme extracellular 833:845 arg1 matrix 847:852 extracellular matrix 833:852 extracellular matrix 833:852 The conformation of the prodomain integrin-binding motif differs in the presence and absence of integrin binding; differences extend well outside the interface and illustrate how integrins can remodel extracellular matrix. 28117447 4 44 theme TGF-β 612:616 arg1 release 601:607 force-dependent release 585:607 force-dependent release of TGF-β from latency 585:629 Here we show how integrin αVβ6 binds pro-TGF-β1 in an orientation biologically relevant for force-dependent release of TGF-β from latency. 28117447 0 45 theme TGF-β 63:67 arg1 activation 49:58 activation 49:58 activation of TGF-β 49:67 Force interacts with macromolecular structure in activation of TGF-β. 28117447 2 46 theme precursor 259:267 arg1 activation 211:220 activation 211:220 activation of the transforming growth factor-β1 precursor (pro-TGF-β1) 211:280 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 28117447 2 47 theme growth 351:356 arg1 factor 358:363 the TGF-β growth factor 341:363 the TGF-β growth factor 341:363 In activation of the transforming growth factor-β1 precursor (pro-TGF-β1), integrins bind to the prodomain, apply force, and release the TGF-β growth factor. 17567119 6 0 theme inhibitors 1295:1304 arg1 design 1275:1280 the rational structure-based drug design 1241:1280 the rational structure-based drug design of new GCPII inhibitors in the future 1241:1318 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 1 1 from neuroprotective 162:176 arg1 models 202:207 multiple preclinical models 181:207 multiple preclinical models in which dysregulated glutamatergic transmission is implicated 181:270 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 6 2 theme new 1285:1287 arg1 inhibitors 1295:1304 new GCPII inhibitors 1285:1304 new GCPII inhibitors 1285:1304 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 5 3 theme structural 1151:1160 arg1 features 1162:1169 the structural features 1147:1169 the structural features required for potent GCPII inhibition 1147:1206 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 6 4 theme GCPII 1289:1293 arg1 inhibitors 1295:1304 new GCPII inhibitors 1285:1304 new GCPII inhibitors 1285:1304 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 3 5 dep S1 690:691 arg1 i.e. 684:687 i.e., S1' 684:692 i.e. 684:687 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 3 6 theme der 772:774 arg1 interactions 782:793 polar and van der Waals interactions 758:793 polar and van der Waals interactions 758:793 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 4 7 theme inhibitors 848:857 arg1 parts 835:839 the distal parts 824:839 the distal parts of the inhibitors 824:857 The structural diversity of the distal parts of the inhibitors leads to rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor. 17567119 3 8 theme similar 640:646 arg1 modes 656:660 similar binding modes 640:660 similar binding modes 640:660 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 3 9 theme Waals 776:780 arg1 interactions 782:793 polar and van der Waals interactions 758:793 polar and van der Waals interactions 758:793 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 3 10 theme pharmacophore 669:681 arg1 pocket 695:700 the pharmacophore (i.e., S1') pocket 665:700 the pharmacophore (i.e., S1') pocket of GCPII 665:709 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 2 11 theme crystal 291:297 arg1 structures 299:308 crystal structures 291:308 crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution 291:518 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 5 12 theme GCPII 1105:1109 arg1 pocket 1125:1130 the GCPII pharmacophore pocket 1101:1130 the GCPII pharmacophore pocket 1101:1130 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 3 13 dep pocket 695:700 arg1 S1 690:691 S1' 690:692 S1' 690:692 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 5 14 with conjunction 1023:1033 arg1 data 1066:1069 the available biochemical data 1040:1069 the available biochemical data 1040:1069 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 6 15 theme drug 1270:1273 arg1 design 1275:1280 the rational structure-based drug design 1241:1280 the rational structure-based drug design of new GCPII inhibitors in the future 1241:1318 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 5 16 theme structures 993:1002 arg1 structures 993:1002 structures 993:1002 structures presented here 993:1017 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 5 16 theme structures 993:1002 arg1 set 986:988 The set 982:988 The set of structures presented here 982:1017 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 4 17 theme site 894:897 arg1 rearrangements 868:881 rearrangements 868:881 rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor 868:979 The structural diversity of the distal parts of the inhibitors leads to rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor. 17567119 5 18 theme pocket 1125:1130 arg1 flexibility 1086:1096 a flexibility 1084:1096 a flexibility of the GCPII pharmacophore pocket 1084:1130 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 2 19 theme pentanedioic 401:412 arg1 2-PMPA 420:425 2-PMPA 420:425 2-PMPA 420:425 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 19 theme pentanedioic 401:412 arg1 acid 414:417 2-(phosphonomethyl)pentanedioic acid 382:417 2-(phosphonomethyl)pentanedioic acid (2-PMPA) 382:426 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 19 theme pentanedioic 401:412 arg1 mimetics/derivatives 360:379 three glutamate mimetics/derivatives 344:379 three glutamate mimetics/derivatives 344:379 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 20 theme GCPII 323:327 arg1 structures 299:308 crystal structures 291:308 crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution 291:518 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 0 21 theme Structural 0:9 arg1 insight 11:17 Structural insight 0:17 Structural insight into the pharmacophore pocket of human glutamate carboxypeptidase II 0:86 Structural insight into the pharmacophore pocket of human glutamate carboxypeptidase II. 17567119 2 22 theme human 317:321 arg1 GCPII 323:327 the human GCPII 313:327 the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution 313:518 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 23 theme phosphonomethyl 385:399 arg1 2-PMPA 420:425 2-PMPA 420:425 2-PMPA 420:425 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 23 theme phosphonomethyl 385:399 arg1 acid 414:417 2-(phosphonomethyl)pentanedioic acid 382:417 2-(phosphonomethyl)pentanedioic acid (2-PMPA) 382:426 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 23 theme phosphonomethyl 385:399 arg1 mimetics/derivatives 360:379 three glutamate mimetics/derivatives 344:379 three glutamate mimetics/derivatives 344:379 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 6 24 from design 1275:1280 arg1 future 1313:1318 future 1313:1318 future 1313:1318 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 4 25 theme distal 828:833 arg1 parts 835:839 the distal parts 824:839 the distal parts of the inhibitors 824:857 The structural diversity of the distal parts of the inhibitors leads to rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor. 17567119 1 26 theme glutamate 103:111 arg1 GCPII 134:138 GCPII 134:138 GCPII 134:138 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 1 26 theme glutamate 103:111 arg1 carboxypeptidase 113:128 glutamate carboxypeptidase II 103:131 glutamate carboxypeptidase II (GCPII) 103:139 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 1 27 theme preclinical 190:200 arg1 models 202:207 multiple preclinical models 181:207 multiple preclinical models in which dysregulated glutamatergic transmission is implicated 181:270 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 2 28 theme quisqualic 429:438 arg1 QA 446:447 QA 446:447 QA 446:447 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 28 theme quisqualic 429:438 arg1 acid 440:443 quisqualic acid 429:443 quisqualic acid (QA) 429:448 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 28 theme quisqualic 429:438 arg1 mimetics/derivatives 360:379 three glutamate mimetics/derivatives 344:379 three glutamate mimetics/derivatives 344:379 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 6 29 theme rational 1245:1252 arg1 design 1275:1280 the rational structure-based drug design 1241:1280 the rational structure-based drug design of new GCPII inhibitors in the future 1241:1318 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 1 30 theme carboxypeptidase 113:128 arg1 Inhibition 89:98 Inhibition 89:98 Inhibition of glutamate carboxypeptidase II (GCPII) 89:139 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 2 31 theme L-serine 455:462 arg1 L-SOS 475:479 L-SOS 475:479 L-SOS 475:479 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 31 theme L-serine 455:462 arg1 O-sulfate 464:472 L-serine O-sulfate 455:472 L-serine O-sulfate (L-SOS) 455:480 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 31 theme L-serine 455:462 arg1 mimetics/derivatives 360:379 three glutamate mimetics/derivatives 344:379 three glutamate mimetics/derivatives 344:379 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 3 32 theme inhibitors 602:611 arg1 parts 589:593 the distal parts 578:593 the distal parts of the inhibitors 578:611 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 1 33 theme multiple 181:188 arg1 models 202:207 multiple preclinical models 181:207 multiple preclinical models in which dysregulated glutamatergic transmission is implicated 181:270 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 0 34 theme pharmacophore 28:40 arg1 pocket 42:47 the pharmacophore pocket 24:47 the pharmacophore pocket of human glutamate carboxypeptidase II 24:86 Structural insight into the pharmacophore pocket of human glutamate carboxypeptidase II. 17567119 4 35 theme parts 835:839 arg1 diversity 811:819 The structural diversity 796:819 The structural diversity of the distal parts of the inhibitors 796:857 The structural diversity of the distal parts of the inhibitors leads to rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor. 17567119 3 36 theme structural 547:556 arg1 differences 558:568 the structural differences 543:568 the structural differences between the distal parts of the inhibitors 543:611 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 0 37 theme human 52:56 arg1 carboxypeptidase 68:83 human glutamate carboxypeptidase II 52:86 human glutamate carboxypeptidase II 52:86 Structural insight into the pharmacophore pocket of human glutamate carboxypeptidase II. 17567119 3 38 theme GCPII 705:709 arg1 pocket 695:700 the pharmacophore (i.e., S1') pocket 665:700 the pharmacophore (i.e., S1') pocket of GCPII 665:709 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 4 39 theme structural 800:809 arg1 diversity 811:819 The structural diversity 796:819 The structural diversity of the distal parts of the inhibitors 796:857 The structural diversity of the distal parts of the inhibitors leads to rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor. 17567119 2 40 theme glutamate 350:358 arg1 O-sulfate 464:472 L-serine O-sulfate 455:472 L-serine O-sulfate (L-SOS) 455:480 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 40 theme glutamate 350:358 arg1 acid 414:417 2-(phosphonomethyl)pentanedioic acid 382:417 2-(phosphonomethyl)pentanedioic acid (2-PMPA) 382:426 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 40 theme glutamate 350:358 arg1 acid 440:443 quisqualic acid 429:443 quisqualic acid (QA) 429:448 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 2 40 theme glutamate 350:358 arg1 mimetics/derivatives 360:379 three glutamate mimetics/derivatives 344:379 three glutamate mimetics/derivatives 344:379 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 5 41 theme available 1044:1052 arg1 data 1066:1069 the available biochemical data 1040:1069 the available biochemical data 1040:1069 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 6 42 theme structure-based 1254:1268 arg1 design 1275:1280 the rational structure-based drug design 1241:1280 the rational structure-based drug design of new GCPII inhibitors in the future 1241:1318 These findings could facilitate the rational structure-based drug design of new GCPII inhibitors in the future. 17567119 5 43 theme biochemical 1054:1064 arg1 data 1066:1069 the available biochemical data 1040:1069 the available biochemical data 1040:1069 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 1 44 from models 202:207 arg1 neuroprotective 162:176 neuroprotective 162:176 neuroprotective 162:176 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 3 45 theme van 768:770 arg1 interactions 782:793 polar and van der Waals interactions 758:793 polar and van der Waals interactions 758:793 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 1 46 theme dysregulated 218:229 arg1 transmission 245:256 dysregulated glutamatergic transmission 218:256 dysregulated glutamatergic transmission 218:256 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 2 47 theme A 507:507 arg1 resolution 509:518 1.72, 1.62, and 2.10 A resolution 486:518 1.72, 1.62, and 2.10 A resolution 486:518 Herein, we report crystal structures of the human GCPII complexed with three glutamate mimetics/derivatives, 2-(phosphonomethyl)pentanedioic acid (2-PMPA), quisqualic acid (QA), and L-serine O-sulfate (L-SOS), at 1.72, 1.62, and 2.10 A resolution, respectively. 17567119 3 48 theme distal 582:587 arg1 parts 589:593 the distal parts 578:593 the distal parts of the inhibitors 578:611 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 5 49 theme potent 1184:1189 arg1 inhibition 1197:1206 potent GCPII inhibition 1184:1206 potent GCPII inhibition 1184:1206 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 5 50 theme GCPII 1191:1195 arg1 inhibition 1197:1206 potent GCPII inhibition 1184:1206 potent GCPII inhibition 1184:1206 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 1 51 theme glutamatergic 231:243 arg1 transmission 245:256 dysregulated glutamatergic transmission 218:256 dysregulated glutamatergic transmission 218:256 Inhibition of glutamate carboxypeptidase II (GCPII) has been shown to be neuroprotective in multiple preclinical models in which dysregulated glutamatergic transmission is implicated. 17567119 0 52 theme carboxypeptidase 68:83 arg1 pocket 42:47 the pharmacophore pocket 24:47 the pharmacophore pocket of human glutamate carboxypeptidase II 24:86 Structural insight into the pharmacophore pocket of human glutamate carboxypeptidase II. 17567119 3 53 theme polar 758:762 arg1 interactions 782:793 polar and van der Waals interactions 758:793 polar and van der Waals interactions 758:793 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 0 54 theme glutamate 58:66 arg1 carboxypeptidase 68:83 human glutamate carboxypeptidase II 52:86 human glutamate carboxypeptidase II 52:86 Structural insight into the pharmacophore pocket of human glutamate carboxypeptidase II. 17567119 3 55 theme binding 648:654 arg1 modes 656:660 similar binding modes 640:660 similar binding modes 640:660 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 17567119 4 56 theme efficient 922:930 arg1 interactions 932:943 efficient interactions 922:943 efficient interactions between the enzyme and an inhibitor 922:979 The structural diversity of the distal parts of the inhibitors leads to rearrangements of the S1' site that are necessary for efficient interactions between the enzyme and an inhibitor. 17567119 5 57 theme pharmacophore 1111:1123 arg1 pocket 1125:1130 the GCPII pharmacophore pocket 1101:1130 the GCPII pharmacophore pocket 1101:1130 The set of structures presented here, in conjunction with the available biochemical data, illustrates a flexibility of the GCPII pharmacophore pocket and highlights the structural features required for potent GCPII inhibition. 17567119 3 58 theme interactions 782:793 arg1 combination 743:753 a combination 741:753 a combination of polar and van der Waals interactions 741:793 Despite the structural differences between the distal parts of the inhibitors, all three compounds share similar binding modes in the pharmacophore (i.e., S1') pocket of GCPII, where they are stabilized by a combination of polar and van der Waals interactions. 18234225 7 0 theme GCPII 1428:1432 arg1 preference 1434:1443 GCPII preference 1428:1443 GCPII preference 1428:1443 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 9 1 theme structural 1868:1877 arg1 analysis 1879:1886 the structural analysis 1864:1886 the structural analysis 1864:1886 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 7 2 theme P1 1358:1359 arg1 moiety 1361:1366 a P1 moiety 1356:1366 a P1 moiety of a substrate/inhibitor 1356:1391 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 10 3 theme solid 1923:1927 arg1 foundation 1929:1938 an a solid foundation 1918:1938 an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents 1918:2038 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 1 4 theme natural 263:269 arg1 N-acetyl-aspartyl-glutamate 283:309 N-acetyl-aspartyl-glutamate 283:309 N-acetyl-aspartyl-glutamate 283:309 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 1 4 theme natural 263:269 arg1 substrates 271:280 two natural substrates 259:280 two natural substrates 259:280 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 1 4 theme natural 263:269 arg1 folyl-poly-gamma-glutamates 315:341 folyl-poly-gamma-glutamates 315:341 folyl-poly-gamma-glutamates 315:341 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 4 5 theme structural 651:660 arg1 characterization 662:677 structural characterization 651:677 structural characterization of GCPII to its S1 site 651:701 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 6 6 theme Arg536 1188:1193 arg1 importance 1146:1155 the importance 1142:1155 the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues 1142:1258 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 1 7 theme signal 172:177 arg1 transduction 179:190 neuronal signal transduction 163:190 neuronal signal transduction 163:190 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 5 8 theme crystal 827:833 arg1 structures 835:844 crystal structures 827:844 crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 827:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 6 9 dep P1 1237:1238 arg1 i.e. 1231:1234 i.e. 1231:1234 i.e. 1231:1234 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 6 10 theme first 1108:1112 arg1 time 1114:1117 the first time 1104:1117 the first time 1104:1117 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 6 11 theme residues 1251:1258 arg1 recognition 1199:1209 recognition 1199:1209 recognition of the penultimate (i.e., P1) substrate residues 1199:1258 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 1 12 theme folate 207:212 arg1 absorption 214:223 intestinal folate absorption 196:223 intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively 196:355 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 7 13 theme guanidinium 1312:1322 arg1 Arg536 1345:1350 Arg536 1345:1350 Arg536 1345:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 13 theme guanidinium 1312:1322 arg1 Arg534 1334:1339 Arg534 1334:1339 Arg534 1334:1339 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 13 theme guanidinium 1312:1322 arg1 groups 1324:1329 the positively charged guanidinium groups 1289:1329 the positively charged guanidinium groups of Arg534 and Arg536 1289:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 10 14 theme novel 1977:1981 arg1 inhibitors 2010:2019 novel low-molecular-weight GCPII inhibitors 1977:2019 novel low-molecular-weight GCPII inhibitors 1977:2019 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 6 15 theme Arg463 1168:1173 arg1 importance 1146:1155 the importance 1142:1155 the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues 1142:1258 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 5 16 theme phosphapeptide 877:890 arg1 analogs 892:898 phosphapeptide analogs 877:898 phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 877:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 9 17 theme GCPII 1722:1726 arg1 derivatives 1738:1748 several GCPII substrate derivatives 1714:1748 several GCPII substrate derivatives modified at the P1 position, also included in this report, 1714:1807 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 4 18 theme dipeptide-based 712:726 arg1 inhibitors 728:737 dipeptide-based inhibitors 712:737 dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme 712:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 5 19 theme aspartyl-glutamate 926:943 arg1 analogs 892:898 phosphapeptide analogs 877:898 phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 877:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 5 20 from structures 835:844 arg1 complex 864:870 complex 864:870 complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 864:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 9 21 theme derivatives 1738:1748 arg1 hydrolysis 1700:1709 the hydrolysis 1696:1709 the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, 1696:1807 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 7 22 theme Direct 1261:1266 arg1 interactions 1268:1279 Direct interactions 1261:1279 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 1261:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 10 23 theme GCPII 2004:2008 arg1 inhibitors 2010:2019 novel low-molecular-weight GCPII inhibitors 1977:2019 novel low-molecular-weight GCPII inhibitors 1977:2019 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 5 24 theme GCPII 855:859 arg1 structures 835:844 crystal structures 827:844 crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 827:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 4 25 theme S1 695:696 arg1 site 698:701 its S1 site 691:701 its S1 site 691:701 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 9 26 theme P1 1766:1767 arg1 position 1769:1776 the P1 position 1762:1776 the P1 position 1762:1776 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 4 27 theme S1 763:764 arg1 sites 774:778 both S1 and S1' sites 758:778 both S1 and S1' sites of the enzyme 758:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 4 27 theme S1 763:764 arg1 enzyme 787:792 the enzyme 783:792 the enzyme 783:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 10 28 theme agents 2033:2038 arg1 design 1967:1972 the structurally aided design 1944:1972 the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents 1944:2038 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 5 29 theme gamma-glutamyl-glutamate 950:973 arg1 analogs 892:898 phosphapeptide analogs 877:898 phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 877:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 4 30 theme S1 770:771 arg1 sites 774:778 both S1 and S1' sites 758:778 both S1 and S1' sites of the enzyme 758:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 4 30 theme S1 770:771 arg1 enzyme 787:792 the enzyme 783:792 the enzyme 783:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 0 31 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 0:102 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 18234225 1 32 theme substrates 271:280 arg1 hydrolysis 241:250 the hydrolysis 237:250 the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates 237:341 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 8 33 theme conformational 1491:1504 arg1 flexibility 1506:1516 observed conformational flexibility 1482:1516 observed conformational flexibility of the Arg463 and Arg536 side chains 1482:1553 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 2 34 theme GCPII 449:453 arg1 analysis 437:444 the structural analysis 422:444 the structural analysis of GCPII 422:453 During the past years, tremendous efforts have been made toward the structural analysis of GCPII. 18234225 1 35 theme glutamate 110:118 arg1 GCPII 141:145 Human glutamate carboxypeptidase II (GCPII) 104:146 Human glutamate carboxypeptidase II (GCPII) 104:146 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 3 36 theme GCPII 478:482 arg1 structures 464:473 Crystal structures 456:473 Crystal structures of GCPII in complex with various ligands 456:514 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 8 37 theme Arg463 1525:1530 arg1 flexibility 1506:1516 observed conformational flexibility 1482:1516 observed conformational flexibility of the Arg463 and Arg536 side chains 1482:1553 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 9 38 attach derived 1851:1857 arg1 analysis 1879:1886 the structural analysis 1864:1886 the structural analysis 1864:1886 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 9 38 attach derived 1851:1857 arg2 conclusions 1839:1849 conclusions 1839:1849 conclusions derived from the structural analysis 1839:1886 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 5 39 dep determined 816:825 arg1 refined 976:982 refined 976:982 refined 976:982 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 0 40 theme glutamate 47:55 arg1 carboxypeptidase 57:72 human glutamate carboxypeptidase II 41:75 human glutamate carboxypeptidase II 41:75 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 18234225 7 41 theme acidic 1449:1454 arg1 dipeptides 1456:1465 acidic dipeptides 1449:1465 acidic dipeptides 1449:1465 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 5 42 theme A 1008:1008 arg1 resolution 1010:1019 1.50, 1.60, and 1.67 A resolution 987:1019 1.50, 1.60, and 1.67 A resolution 987:1019 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 8 43 theme Arg536 1536:1541 arg1 chains 1548:1553 the Arg463 and Arg536 side chains 1521:1553 chains 1548:1553 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 3 44 theme various 500:506 arg1 ligands 508:514 various ligands 500:514 various ligands 500:514 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 7 45 theme preference 1434:1443 arg1 explanation 1413:1423 mechanistic explanation 1401:1423 mechanistic explanation of GCPII preference for acidic dipeptides 1401:1465 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 8 46 theme chains 1548:1553 arg1 flexibility 1506:1516 observed conformational flexibility 1482:1516 observed conformational flexibility of the Arg463 and Arg536 side chains 1482:1553 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 8 47 theme GCPII 1629:1633 arg1 specificity 1645:1655 GCPII substrate specificity 1629:1655 GCPII substrate specificity 1629:1655 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 6 48 theme penultimate 1218:1228 arg1 P1 1237:1238 P1 1237:1238 P1 1237:1238 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 6 48 theme penultimate 1218:1228 arg1 residues 1251:1258 the penultimate (i.e., P1) substrate residues 1214:1258 the penultimate (i.e., P1) substrate residues 1214:1258 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 2 49 theme tremendous 381:390 arg1 efforts 392:398 tremendous efforts 381:398 tremendous efforts 381:398 During the past years, tremendous efforts have been made toward the structural analysis of GCPII. 18234225 7 50 theme mechanistic 1401:1411 arg1 explanation 1413:1423 mechanistic explanation 1401:1423 mechanistic explanation of GCPII preference for acidic dipeptides 1401:1465 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 6 51 theme GCPII 1053:1057 arg1 pocket 1043:1048 The S1 pocket 1036:1048 The S1 pocket of GCPII 1036:1057 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 3 52 theme ligands 564:570 arg1 binding 547:553 the binding 543:553 the binding of these ligands, particularly to the S1' site of the enzyme 543:614 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 7 53 theme substrate/inhibitor 1373:1391 arg1 interactions 1268:1279 Direct interactions 1261:1279 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 1261:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 53 theme substrate/inhibitor 1373:1391 arg1 moiety 1361:1366 a P1 moiety 1356:1366 a P1 moiety of a substrate/inhibitor 1356:1391 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 8 54 theme different 1594:1602 arg1 inhibitors 1604:1613 different inhibitors 1594:1613 different inhibitors 1594:1613 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 10 55 theme a 1921:1921 arg1 foundation 1929:1938 an a solid foundation 1918:1938 an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents 1918:2038 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 1 56 theme neuronal 163:170 arg1 transduction 179:190 neuronal signal transduction 163:190 neuronal signal transduction 163:190 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 9 57 theme biochemical 1662:1672 arg1 experiments 1674:1684 The biochemical experiments 1658:1684 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, 1658:1807 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 7 58 theme Arg536 1345:1350 arg1 Arg536 1345:1350 Arg536 1345:1350 Arg536 1345:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 58 theme Arg536 1345:1350 arg1 Arg534 1334:1339 Arg534 1334:1339 Arg534 1334:1339 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 58 theme Arg536 1345:1350 arg1 groups 1324:1329 the positively charged guanidinium groups 1289:1329 the positively charged guanidinium groups of Arg534 and Arg536 1289:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 1 59 theme intestinal 196:205 arg1 absorption 214:223 intestinal folate absorption 196:223 intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively 196:355 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 7 60 theme Arg534 1334:1339 arg1 Arg536 1345:1350 Arg536 1345:1350 Arg536 1345:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 60 theme Arg534 1334:1339 arg1 Arg534 1334:1339 Arg534 1334:1339 Arg534 1334:1339 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 60 theme Arg534 1334:1339 arg1 groups 1324:1329 the positively charged guanidinium groups 1289:1329 the positively charged guanidinium groups of Arg534 and Arg536 1289:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 3 61 with complex 487:493 arg1 ligands 508:514 various ligands 500:514 various ligands 500:514 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 1 62 theme carboxypeptidase 120:135 arg1 GCPII 141:145 Human glutamate carboxypeptidase II (GCPII) 104:146 Human glutamate carboxypeptidase II (GCPII) 104:146 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 9 63 theme several 1714:1720 arg1 derivatives 1738:1748 several GCPII substrate derivatives 1714:1748 several GCPII substrate derivatives modified at the P1 position, also included in this report, 1714:1807 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 7 64 theme charged 1304:1310 arg1 Arg536 1345:1350 Arg536 1345:1350 Arg536 1345:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 64 theme charged 1304:1310 arg1 Arg534 1334:1339 Arg534 1334:1339 Arg534 1334:1339 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 7 64 theme charged 1304:1310 arg1 groups 1324:1329 the positively charged guanidinium groups 1289:1329 the positively charged guanidinium groups of Arg534 and Arg536 1289:1350 Direct interactions between the positively charged guanidinium groups of Arg534 and Arg536 and a P1 moiety of a substrate/inhibitor provide mechanistic explanation of GCPII preference for acidic dipeptides. 18234225 10 65 theme aided 1961:1965 arg1 design 1967:1972 the structurally aided design 1944:1972 the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents 1944:2038 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 9 66 theme substrate 1728:1736 arg1 derivatives 1738:1748 several GCPII substrate derivatives 1714:1748 several GCPII substrate derivatives modified at the P1 position, also included in this report, 1714:1807 The biochemical experiments assessing the hydrolysis of several GCPII substrate derivatives modified at the P1 position, also included in this report, further complement and extend conclusions derived from the structural analysis. 18234225 10 67 theme inhibitors 2010:2019 arg1 design 1967:1972 the structurally aided design 1944:1972 the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents 1944:2038 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 3 68 theme enzyme 609:614 arg1 site 597:600 the S1' site 589:600 the S1' site of the enzyme 589:614 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 3 68 theme enzyme 609:614 arg1 enzyme 609:614 the enzyme 605:614 the enzyme 605:614 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 5 69 theme human 849:853 arg1 GCPII 855:859 human GCPII 849:859 human GCPII 849:859 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 4 70 theme GCPII 682:686 arg1 characterization 662:677 structural characterization 651:677 structural characterization of GCPII to its S1 site 651:701 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 10 71 theme low-molecular-weight 1983:2002 arg1 inhibitors 2010:2019 novel low-molecular-weight GCPII inhibitors 1977:2019 novel low-molecular-weight GCPII inhibitors 1977:2019 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 0 72 theme substrate 85:93 arg1 analogs 95:101 its substrate analogs 81:101 its substrate analogs 81:101 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 18234225 5 73 with complex 864:870 arg1 analogs 892:898 phosphapeptide analogs 877:898 phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 877:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 10 74 theme imaging 2025:2031 arg1 agents 2033:2038 imaging agents 2025:2038 imaging agents 2025:2038 The data described here form an a solid foundation for the structurally aided design of novel low-molecular-weight GCPII inhibitors and imaging agents. 18234225 8 75 theme observed 1482:1489 arg1 flexibility 1506:1516 observed conformational flexibility 1482:1516 observed conformational flexibility of the Arg463 and Arg536 side chains 1482:1553 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 5 76 theme folyl-gamma-glutamate 903:923 arg1 analogs 892:898 phosphapeptide analogs 877:898 phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate 877:973 To this end, we have determined crystal structures of human GCPII in complex with phosphapeptide analogs of folyl-gamma-glutamate, aspartyl-glutamate, and gamma-glutamyl-glutamate, refined at 1.50, 1.60, and 1.67 A resolution, respectively. 18234225 6 77 theme S1 1040:1041 arg1 pocket 1043:1048 The S1 pocket 1036:1048 The S1 pocket of GCPII 1036:1057 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 6 78 theme substrate 1241:1249 arg1 P1 1237:1238 P1 1237:1238 P1 1237:1238 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 6 78 theme substrate 1241:1249 arg1 residues 1251:1258 the penultimate (i.e., P1) substrate residues 1214:1258 the penultimate (i.e., P1) substrate residues 1214:1258 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 3 79 theme Crystal 456:462 arg1 structures 464:473 Crystal structures 456:473 Crystal structures of GCPII in complex with various ligands 456:514 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 18234225 0 80 theme interactions 20:31 arg1 basis 11:15 Structural basis 0:15 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 0:102 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 18234225 1 81 theme Human 104:108 arg1 GCPII 141:145 Human glutamate carboxypeptidase II (GCPII) 104:146 Human glutamate carboxypeptidase II (GCPII) 104:146 Human glutamate carboxypeptidase II (GCPII) is involved in neuronal signal transduction and intestinal folate absorption by means of the hydrolysis of its two natural substrates, N-acetyl-aspartyl-glutamate and folyl-poly-gamma-glutamates, respectively. 18234225 6 82 theme Arg534 1176:1181 arg1 importance 1146:1155 the importance 1142:1155 the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues 1142:1258 The S1 pocket of GCPII could be accurately defined and analyzed for the first time, and the data indicate the importance of Asn519, Arg463, Arg534, and Arg536 for recognition of the penultimate (i.e., P1) substrate residues. 18234225 0 83 theme human 41:45 arg1 carboxypeptidase 57:72 human glutamate carboxypeptidase II 41:75 human glutamate carboxypeptidase II 41:75 Structural basis of interactions between human glutamate carboxypeptidase II and its substrate analogs. 18234225 4 84 theme enzyme 787:792 arg1 sites 774:778 both S1 and S1' sites 758:778 both S1 and S1' sites of the enzyme 758:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 4 84 theme enzyme 787:792 arg1 enzyme 787:792 the enzyme 783:792 the enzyme 783:792 In this article, we have extended structural characterization of GCPII to its S1 site by using dipeptide-based inhibitors that interact with both S1 and S1' sites of the enzyme. 18234225 2 85 theme structural 426:435 arg1 analysis 437:444 the structural analysis 422:444 the structural analysis of GCPII 422:453 During the past years, tremendous efforts have been made toward the structural analysis of GCPII. 18234225 8 86 theme side 1543:1546 arg1 chains 1548:1553 the Arg463 and Arg536 side chains 1521:1553 chains 1548:1553 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 2 87 theme past 369:372 arg1 years 374:378 the past years 365:378 the past years 365:378 During the past years, tremendous efforts have been made toward the structural analysis of GCPII. 18234225 8 88 theme substrate 1635:1643 arg1 specificity 1645:1655 GCPII substrate specificity 1629:1655 GCPII substrate specificity 1629:1655 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 8 89 theme GCPII 1572:1576 arg1 affinity 1578:1585 GCPII affinity 1572:1585 GCPII affinity toward different inhibitors 1572:1613 Additionally, observed conformational flexibility of the Arg463 and Arg536 side chains likely regulates GCPII affinity toward different inhibitors and modulates GCPII substrate specificity. 18234225 3 90 from structures 464:473 arg1 complex 487:493 complex 487:493 complex with various ligands 487:514 Crystal structures of GCPII in complex with various ligands have provided insight into the binding of these ligands, particularly to the S1' site of the enzyme. 17372356 3 0 with complex 559:565 arg1 inhibitors 583:592 two potent inhibitors 572:592 two potent inhibitors 572:592 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 0 with complex 559:565 arg1 2-PMPA 611:616 2-PMPA 611:616 2-PMPA 611:616 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 0 with complex 559:565 arg1 quisqualate 595:605 quisqualate 595:605 quisqualate 595:605 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 2 1 dep detection 404:412 arg1 the 400:402 the 400:402 the 400:402 Inhibitors of the enzyme have been shown to provide neuroprotection, but may also be useful for the detection, imaging and treatment of prostate cancer. 17372356 5 2 theme bridges 991:997 arg1 denominator 901:911 The common denominator 890:911 The common denominator for high-affinity binding to the S1' site 890:953 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 5 2 theme bridges 991:997 arg1 formation 962:970 the formation 958:970 the formation of two strong salt bridges 958:997 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 4 3 theme inhibitors 762:771 arg1 binding 747:753 binding 747:753 binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme 747:887 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 5 4 theme salt 986:989 arg1 bridges 991:997 two strong salt bridges 975:997 two strong salt bridges 975:997 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 1 5 theme human 226:230 arg1 prostate 232:239 human prostate 226:239 human prostate (where it is called prostate-specific membrane antigen; PSMA) 226:301 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 0 6 theme potent 88:93 arg1 2-PMPA 123:128 2-PMPA 123:128 2-PMPA 123:128 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 0 6 theme potent 88:93 arg1 quisqualate 107:117 quisqualate 107:117 quisqualate 107:117 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 0 6 theme potent 88:93 arg1 inhibitors 95:104 two potent inhibitors 84:104 two potent inhibitors 84:104 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 3 7 theme strongest 623:631 arg1 inhibitor 639:647 the strongest GCPII inhibitor 619:647 the strongest GCPII inhibitor to date 619:655 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 7 theme strongest 623:631 arg1 quisqualate 595:605 quisqualate 595:605 quisqualate 595:605 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 0 8 with complex 71:77 arg1 2-PMPA 123:128 2-PMPA 123:128 2-PMPA 123:128 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 0 8 with complex 71:77 arg1 quisqualate 107:117 quisqualate 107:117 quisqualate 107:117 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 0 8 with complex 71:77 arg1 inhibitors 95:104 two potent inhibitors 84:104 two potent inhibitors 84:104 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 1 9 dep antigen 288:294 arg1 PSMA 297:300 PSMA 297:300 PSMA 297:300 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 3 10 theme A 689:689 arg1 resolutions 662:672 resolutions 662:672 resolutions of 3.0 and 2.2 A 662:689 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 0 11 from structures 48:57 arg1 complex 71:77 complex 71:77 complex with two potent inhibitors, quisqualate and 2-PMPA 71:128 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 2 12 theme cancer 449:454 arg1 detection 404:412 detection 404:412 detection 404:412 Inhibitors of the enzyme have been shown to provide neuroprotection, but may also be useful for the detection, imaging and treatment of prostate cancer. 17372356 2 12 theme cancer 449:454 arg1 treatment 427:435 treatment 427:435 treatment 427:435 Inhibitors of the enzyme have been shown to provide neuroprotection, but may also be useful for the detection, imaging and treatment of prostate cancer. 17372356 2 12 theme cancer 449:454 arg1 imaging 415:421 imaging 415:421 imaging 415:421 Inhibitors of the enzyme have been shown to provide neuroprotection, but may also be useful for the detection, imaging and treatment of prostate cancer. 17372356 5 13 theme common 894:899 arg1 denominator 901:911 The common denominator 890:911 The common denominator for high-affinity binding to the S1' site 890:953 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 5 13 theme common 894:899 arg1 formation 962:970 the formation 958:970 the formation of two strong salt bridges 958:997 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 4 14 theme enzyme 882:887 arg1 site 870:873 the S1' site 862:873 the S1' site of the enzyme 862:887 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 4 14 theme enzyme 882:887 arg1 enzyme 882:887 the enzyme 878:887 the enzyme 878:887 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 0 15 theme glutamate 6:14 arg1 inhibition 36:45 Human glutamate carboxypeptidase II inhibition 0:45 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 0:129 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 0 16 theme Human 0:4 arg1 inhibition 36:45 Human glutamate carboxypeptidase II inhibition 0:45 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 0:129 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 3 17 theme GCPII 521:525 arg1 part 513:516 the extracellular part 495:516 the extracellular part of GCPII 495:525 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 18 theme 3.0 677:679 arg1 A 689:689 3.0 and 2.2 A 677:689 A 689:689 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 19 theme amino-acid 528:537 arg1 residues 539:546 amino-acid residues 528:546 amino-acid residues 44-750 528:553 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 20 theme extracellular 499:511 arg1 part 513:516 the extracellular part 495:516 the extracellular part of GCPII 495:525 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 0 21 theme carboxypeptidase 16:31 arg1 inhibition 36:45 Human glutamate carboxypeptidase II inhibition 0:45 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 0:129 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 3 22 theme Crystal 457:463 arg1 structures 465:474 Crystal structures 457:474 Crystal structures 457:474 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 1 23 theme central 189:195 arg1 system 205:210 the central nervous system 185:210 the central nervous system 185:210 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 1 24 theme prostate-specific 261:277 arg1 antigen 288:294 prostate-specific membrane antigen 261:294 prostate-specific membrane antigen; PSMA) 261:301 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 5 25 theme strong 979:984 arg1 bridges 991:997 two strong salt bridges 975:997 two strong salt bridges 975:997 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 1 26 theme nervous 197:203 arg1 system 205:210 the central nervous system 185:210 the central nervous system 185:210 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 1 27 theme membrane 279:286 arg1 antigen 288:294 prostate-specific membrane antigen 261:294 prostate-specific membrane antigen; PSMA) 261:301 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 4 28 theme L-2-amino-4-phosphonobutanoic 801:829 arg1 acid 831:834 L-2-amino-4-phosphonobutanoic acid 801:834 L-2-amino-4-phosphonobutanoic acid 801:834 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 4 28 theme L-2-amino-4-phosphonobutanoic 801:829 arg1 inhibitors 762:771 the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate 758:857 the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate 758:857 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 3 29 theme potent 576:581 arg1 inhibitors 583:592 two potent inhibitors 572:592 two potent inhibitors 572:592 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 29 theme potent 576:581 arg1 2-PMPA 611:616 2-PMPA 611:616 2-PMPA 611:616 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 29 theme potent 576:581 arg1 quisqualate 595:605 quisqualate 595:605 quisqualate 595:605 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 4 30 dep inhibitors 762:771 arg1 acid 831:834 L-2-amino-4-phosphonobutanoic acid 801:834 L-2-amino-4-phosphonobutanoic acid 801:834 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 4 30 dep inhibitors 762:771 arg1 willardiine 773:783 willardiine 773:783 willardiine 773:783 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 4 30 dep inhibitors 762:771 arg1 inhibitors 762:771 the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate 758:857 the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate 758:857 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 4 30 dep inhibitors 762:771 arg1 L-serine-O-sulfate 840:857 L-serine-O-sulfate 840:857 L-serine-O-sulfate 840:857 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 4 30 dep inhibitors 762:771 arg1 homoibotenate 786:798 homoibotenate 786:798 homoibotenate 786:798 In addition, models were constructed for binding of the inhibitors willardiine, homoibotenate, L-2-amino-4-phosphonobutanoic acid and L-serine-O-sulfate to the S1' site of the enzyme. 17372356 0 31 dep inhibition 36:45 arg1 structures 48:57 structures 48:57 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 0:129 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 3 32 dep determined 481:490 arg1 residues 539:546 amino-acid residues 528:546 amino-acid residues 44-750 528:553 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 33 theme 2.2 685:687 arg1 A 689:689 3.0 and 2.2 A 677:689 A 689:689 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 1 34 theme glutamate 137:145 arg1 carboxypeptidase 147:162 Human glutamate carboxypeptidase II 131:165 Human glutamate carboxypeptidase II (GCPII) 131:173 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 1 34 theme glutamate 137:145 arg1 GCPII 168:172 GCPII 168:172 GCPII 168:172 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 2 35 theme prostate 440:447 arg1 cancer 449:454 prostate cancer 440:454 prostate cancer 440:454 Inhibitors of the enzyme have been shown to provide neuroprotection, but may also be useful for the detection, imaging and treatment of prostate cancer. 17372356 5 36 theme high-affinity 917:929 arg1 binding 931:937 high-affinity binding 917:937 high-affinity binding to the S1' site 917:953 The common denominator for high-affinity binding to the S1' site is the formation of two strong salt bridges. 17372356 1 37 theme Human 131:135 arg1 carboxypeptidase 147:162 Human glutamate carboxypeptidase II 131:165 Human glutamate carboxypeptidase II (GCPII) 131:173 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 1 37 theme Human 131:135 arg1 GCPII 168:172 GCPII 168:172 GCPII 168:172 Human glutamate carboxypeptidase II (GCPII) occurs in the central nervous system as well as in human prostate (where it is called prostate-specific membrane antigen; PSMA). 17372356 0 38 theme GCPII 62:66 arg1 structures 48:57 structures 48:57 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 0:129 Human glutamate carboxypeptidase II inhibition: structures of GCPII in complex with two potent inhibitors, quisqualate and 2-PMPA. 17372356 3 39 theme GCPII 633:637 arg1 inhibitor 639:647 the strongest GCPII inhibitor 619:647 the strongest GCPII inhibitor to date 619:655 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 3 39 theme GCPII 633:637 arg1 quisqualate 595:605 quisqualate 595:605 quisqualate 595:605 Crystal structures were determined of the extracellular part of GCPII (amino-acid residues 44-750) in complex with two potent inhibitors, quisqualate and 2-PMPA (the strongest GCPII inhibitor to date), at resolutions of 3.0 and 2.2 A, respectively. 17372356 2 40 theme enzyme 322:327 arg1 Inhibitors 304:313 Inhibitors 304:313 Inhibitors of the enzyme 304:327 Inhibitors of the enzyme have been shown to provide neuroprotection, but may also be useful for the detection, imaging and treatment of prostate cancer. 17954916 8 0 theme rat 951:953 arg1 interchangeability 919:936 interchangeability 919:936 interchangeability of human and rat IF receptors 919:966 A possible basis for the lack of interchangeability of human and rat IF receptors is presented. 17954916 6 1 theme Cbl 744:746 arg1 trans-Cbl-B12 775:787 trans-Cbl-B12 775:787 trans-Cbl-B12 775:787 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 6 1 theme Cbl 744:746 arg1 complex 766:772 another Cbl transport protein complex 736:772 another Cbl transport protein complex 736:772 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 1 2 with complex 126:132 arg1 Cbl 150:152 Cbl 150:152 Cbl 150:152 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) was determined at 2.6-A resolution. 17954916 1 2 with complex 126:132 arg1 cobalamin 139:147 cobalamin 139:147 cobalamin (Cbl) 139:153 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) was determined at 2.6-A resolution. 17954916 3 3 from interface 320:328 arg1 conformation 358:369 a base-on conformation 348:369 a base-on conformation 348:369 It is a two-domain protein, and the Cbl is bound at the interface of the domains in a base-on conformation. 17954916 4 4 theme asymmetric 549:558 arg1 unit 560:563 the same asymmetric unit 540:563 the same asymmetric unit 540:563 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 0 5 theme 2.6-A 66:70 arg1 resolution 72:81 2.6-A resolution 66:81 2.6-A resolution 66:81 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 4 6 theme full-length 390:400 arg1 each 413:416 each 413:416 each 413:416 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 6 theme full-length 390:400 arg1 molecules 402:410 two full-length molecules 386:410 two full-length molecules 386:410 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 7 located present 529:535 arg2 molecules 402:410 two full-length molecules 386:410 two full-length molecules 386:410 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 7 located present 529:535 arg2 each 413:416 each 413:416 each 413:416 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 7 located present 529:535 arg1 unit 560:563 the same asymmetric unit 540:563 the same asymmetric unit 540:563 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 7 located present 529:535 arg2 molecules 488:496 two truncated molecules 474:496 two truncated molecules with only an alpha- domain 474:523 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 8 with molecules 402:410 arg1 domain 518:523 only an alpha- domain 503:523 only an alpha- domain 503:523 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 9 from present 529:535 arg1 unit 560:563 the same asymmetric unit 540:563 the same asymmetric unit 540:563 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 3 10 theme base-on 350:356 arg1 conformation 358:369 a base-on conformation 348:369 a base-on conformation 348:369 It is a two-domain protein, and the Cbl is bound at the interface of the domains in a base-on conformation. 17954916 4 11 theme same 544:547 arg1 unit 560:563 the same asymmetric unit 540:563 the same asymmetric unit 540:563 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 6 12 theme IF-B12 717:722 arg1 complex 724:730 the IF-B12 complex 713:730 the IF-B12 complex 713:730 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 4 13 theme alpha- 511:516 arg1 domain 518:523 only an alpha- domain 503:523 only an alpha- domain 503:523 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 0 14 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 0:82 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 7 15 from effect 812:817 arg1 binding 826:832 the binding 822:832 the binding of Cbl analogues in transport proteins 822:871 The pH effect on the binding of Cbl analogues in transport proteins is analyzed. 17954916 0 16 theme human 21:25 arg1 factor 37:42 human intrinsic factor 21:42 human intrinsic factor 21:42 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 8 17 theme possible 888:895 arg1 basis 897:901 A possible basis 886:901 A possible basis for the lack of interchangeability of human and rat IF receptors 886:966 A possible basis for the lack of interchangeability of human and rat IF receptors is presented. 17954916 5 18 theme sixth 637:641 arg1 empty 676:680 empty 676:680 empty 676:680 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 5 18 theme sixth 637:641 arg1 position 654:661 the sixth coordinate position 633:661 the sixth coordinate position of Co(2+) 633:671 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 3 19 theme domains 337:343 arg1 interface 320:328 the interface 316:328 the interface of the domains in a base-on conformation 316:369 It is a two-domain protein, and the Cbl is bound at the interface of the domains in a base-on conformation. 17954916 3 20 theme two-domain 272:281 arg1 protein 283:289 a two-domain protein 270:289 a two-domain protein 270:289 It is a two-domain protein, and the Cbl is bound at the interface of the domains in a base-on conformation. 17954916 3 20 theme two-domain 272:281 arg1 It 264:265 It 264:265 It 264:265 It is a two-domain protein, and the Cbl is bound at the interface of the domains in a base-on conformation. 17954916 0 21 from resolution 72:81 arg1 complex 55:61 cobalamin complex 45:61 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 0:82 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 2 22 theme alpha 238:242 arg1 barrel 256:261 an alpha(6)/alpha(6) barrel 235:261 an alpha(6)/alpha(6) barrel 235:261 The overall fold of the molecule is that of an alpha(6)/alpha(6) barrel. 17954916 0 23 theme factor 37:42 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 0:82 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 6 24 theme detailed 685:692 arg1 comparison 694:703 A detailed comparison 683:703 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, 683:788 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 2 25 theme overall 195:201 arg1 that 227:230 that 227:230 that 227:230 The overall fold of the molecule is that of an alpha(6)/alpha(6) barrel. 17954916 2 25 theme overall 195:201 arg1 fold 203:206 The overall fold 191:206 The overall fold of the molecule 191:222 The overall fold of the molecule is that of an alpha(6)/alpha(6) barrel. 17954916 7 26 from binding 826:832 arg1 proteins 864:871 transport proteins 854:871 transport proteins 854:871 The pH effect on the binding of Cbl analogues in transport proteins is analyzed. 17954916 0 27 theme intrinsic 27:35 arg1 factor 37:42 human intrinsic factor 21:42 human intrinsic factor 21:42 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 5 28 theme Co 666:667 arg1 empty 676:680 empty 676:680 empty 676:680 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 5 28 theme Co 666:667 arg1 position 654:661 the sixth coordinate position 633:661 the sixth coordinate position of Co(2+) 633:671 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 1 29 theme 2.6-A 173:177 arg1 resolution 179:188 2.6-A resolution 173:188 2.6-A resolution 173:188 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) was determined at 2.6-A resolution. 17954916 0 30 theme cobalamin 45:53 arg1 complex 55:61 cobalamin complex 45:61 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 0:82 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 4 31 attach present 529:535 arg2 molecules 402:410 two full-length molecules 386:410 two full-length molecules 386:410 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 31 attach present 529:535 arg2 each 413:416 each 413:416 each 413:416 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 31 attach present 529:535 arg1 unit 560:563 the same asymmetric unit 540:563 the same asymmetric unit 540:563 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 31 attach present 529:535 arg2 molecules 488:496 two truncated molecules 474:496 two truncated molecules with only an alpha- domain 474:523 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 8 32 theme interchangeability 919:936 arg1 lack 911:914 the lack 907:914 the lack of interchangeability of human and rat IF receptors 907:966 A possible basis for the lack of interchangeability of human and rat IF receptors is presented. 17954916 1 33 theme intrinsic 101:109 arg1 factor 111:116 intrinsic factor 101:116 intrinsic factor (IF) 101:121 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) was determined at 2.6-A resolution. 17954916 5 34 theme coordinate 643:652 arg1 empty 676:680 empty 676:680 empty 676:680 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 5 34 theme coordinate 643:652 arg1 position 654:661 the sixth coordinate position 633:661 the sixth coordinate position of Co(2+) 633:671 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 7 35 theme transport 854:862 arg1 proteins 864:871 transport proteins 854:871 transport proteins 854:871 The pH effect on the binding of Cbl analogues in transport proteins is analyzed. 17954916 1 36 theme factor 111:116 arg1 structure 88:96 The structure 84:96 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) 84:153 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) was determined at 2.6-A resolution. 17954916 1 37 from structure 88:96 arg1 complex 126:132 complex 126:132 complex with cobalamin (Cbl) 126:153 The structure of intrinsic factor (IF) in complex with cobalamin (Cbl) was determined at 2.6-A resolution. 17954916 6 38 theme protein 758:764 arg1 trans-Cbl-B12 775:787 trans-Cbl-B12 775:787 trans-Cbl-B12 775:787 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 6 38 theme protein 758:764 arg1 complex 766:772 another Cbl transport protein complex 736:772 another Cbl transport protein complex 736:772 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 7 39 theme Cbl 837:839 arg1 analogues 841:849 Cbl analogues 837:849 Cbl analogues 837:849 The pH effect on the binding of Cbl analogues in transport proteins is analyzed. 17954916 2 40 theme molecule 215:222 arg1 that 227:230 that 227:230 that 227:230 The overall fold of the molecule is that of an alpha(6)/alpha(6) barrel. 17954916 2 40 theme molecule 215:222 arg1 fold 203:206 The overall fold 191:206 The overall fold of the molecule 191:222 The overall fold of the molecule is that of an alpha(6)/alpha(6) barrel. 17954916 7 41 theme analogues 841:849 arg1 binding 826:832 the binding 822:832 the binding of Cbl analogues in transport proteins 822:871 The pH effect on the binding of Cbl analogues in transport proteins is analyzed. 17954916 7 42 theme pH 809:810 arg1 effect 812:817 The pH effect 805:817 The pH effect on the binding of Cbl analogues in transport proteins 805:871 The pH effect on the binding of Cbl analogues in transport proteins is analyzed. 17954916 2 43 theme /alpha 246:251 arg1 barrel 256:261 an alpha(6)/alpha(6) barrel 235:261 an alpha(6)/alpha(6) barrel 235:261 The overall fold of the molecule is that of an alpha(6)/alpha(6) barrel. 17954916 8 44 theme human 941:945 arg1 interchangeability 919:936 interchangeability 919:936 interchangeability of human and rat IF receptors 919:966 A possible basis for the lack of interchangeability of human and rat IF receptors is presented. 17954916 4 45 with molecules 488:496 arg1 domain 518:523 only an alpha- domain 503:523 only an alpha- domain 503:523 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 0 46 dep structure 8:16 arg1 complex 55:61 cobalamin complex 45:61 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 0:82 Crystal structure of human intrinsic factor: cobalamin complex at 2.6-A resolution. 17954916 4 47 from unit 560:563 arg1 present 529:535 present 529:535 present 529:535 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 4 48 theme truncated 478:486 arg1 molecules 488:496 two truncated molecules 474:496 two truncated molecules with only an alpha- domain 474:523 Surprisingly, two full-length molecules, each comprising an alpha- and a beta-domain and one Cbl, and two truncated molecules with only an alpha- domain are present in the same asymmetric unit. 17954916 5 49 theme uncharged 609:617 arg1 residues 619:626 uncharged residues 609:626 uncharged residues 609:626 The environment around Cbl is dominated by uncharged residues, and the sixth coordinate position of Co(2+) is empty. 17954916 6 50 theme transport 748:756 arg1 trans-Cbl-B12 775:787 trans-Cbl-B12 775:787 trans-Cbl-B12 775:787 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 17954916 6 50 theme transport 748:756 arg1 complex 766:772 another Cbl transport protein complex 736:772 another Cbl transport protein complex 736:772 A detailed comparison between the IF-B12 complex and another Cbl transport protein complex, trans-Cbl-B12, has been made. 19901337 0 0 theme human 93:97 arg1 receptor 99:106 its human receptor 89:106 its human receptor 89:106 Crystal structure of NL63 respiratory coronavirus receptor-binding domain complexed with its human receptor. 19901337 5 1 theme receptor-binding 743:758 arg1 RBMs 768:771 RBMs 768:771 RBMs 768:771 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 5 1 theme receptor-binding 743:758 arg1 motifs 760:765 3 discontinuous receptor-binding motifs 727:765 3 discontinuous receptor-binding motifs (RBMs) 727:772 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 5 2 theme NL63-CoV 623:630 arg1 RBD 632:634 NL63-CoV RBD 623:634 NL63-CoV RBD 623:634 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 1 3 theme human 150:154 arg1 virus 168:172 a prevalent human respiratory virus 138:172 a prevalent human respiratory virus 138:172 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 3 theme human 150:154 arg1 coronavirus 114:124 NL63 coronavirus 109:124 NL63 coronavirus (NL63-CoV) 109:135 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 7 4 theme I 975:975 arg1 coronaviruses 977:989 group I coronaviruses 969:989 group I coronaviruses 969:989 Among group I coronaviruses, RBD cores are conserved but RBMs are variable, explaining how these viruses recognize different receptors. 19901337 4 5 theme protein 558:564 arg1 RBD 591:593 RBD 591:593 RBD 591:593 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 4 5 theme protein 558:564 arg1 domain 583:588 NL63-CoV spike protein receptor-binding domain 543:588 NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2 543:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 6 6 from " 952:952 arg1 ACE2 957:960 ACE2 957:960 ACE2 957:960 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 8 7 theme viral 1158:1162 arg1 evolution 1164:1172 viral evolution 1158:1172 viral evolution 1158:1172 These results provide a structural basis for understanding viral evolution and virus-receptor interactions. 19901337 2 8 theme group 311:315 arg1 SARS-CoV 338:345 SARS-CoV 338:345 SARS-CoV 338:345 Incidentally, ACE2 is also used by group II SARS coronavirus (SARS-CoV). 19901337 2 8 theme group 311:315 arg1 coronavirus 325:335 group II SARS coronavirus 311:335 group II SARS coronavirus (SARS-CoV) 311:346 Incidentally, ACE2 is also used by group II SARS coronavirus (SARS-CoV). 19901337 4 9 theme spike 552:556 arg1 RBD 591:593 RBD 591:593 RBD 591:593 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 4 9 theme spike 552:556 arg1 domain 583:588 NL63-CoV spike protein receptor-binding domain 543:588 NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2 543:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 4 10 theme domain 583:588 arg1 structure 530:538 the crystal structure 518:538 the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2 518:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 5 11 theme beta-sheets 703:713 arg1 layers 693:698 2 layers 691:698 2 layers of beta-sheets 691:713 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 6 12 contain have 810:813 arg2 homology 829:836 no structural homology 815:836 no structural homology 815:836 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 6 12 contain have 810:813 arg1 SARS-CoV 801:808 SARS-CoV 801:808 SARS-CoV 801:808 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 6 12 contain have 810:813 arg1 NL63-CoV 788:795 NL63-CoV 788:795 NL63-CoV 788:795 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 3 13 theme homologous 440:449 arg1 coronaviruses 459:471 homologous group I coronaviruses 440:471 homologous group I coronaviruses 440:471 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 3 14 theme coronaviruses 389:401 arg1 coronaviruses 389:401 coronaviruses 389:401 coronaviruses 389:401 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 3 14 theme coronaviruses 389:401 arg1 groups 379:384 different groups 369:384 different groups of coronaviruses 369:401 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 4 15 theme receptor-binding 566:581 arg1 RBD 591:593 RBD 591:593 RBD 591:593 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 4 15 theme receptor-binding 566:581 arg1 domain 583:588 NL63-CoV spike protein receptor-binding domain 543:588 NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2 543:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 6 16 theme hotspot 945:951 arg1 " 952:952 a "virus-binding hotspot" 928:952 a "virus-binding hotspot" on ACE2 928:960 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 0 17 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of NL63 respiratory coronavirus receptor-binding domain 0:72 Crystal structure of NL63 respiratory coronavirus receptor-binding domain complexed with its human receptor. 19901337 7 18 theme different 1078:1086 arg1 receptors 1088:1096 different receptors 1078:1096 different receptors 1078:1096 Among group I coronaviruses, RBD cores are conserved but RBMs are variable, explaining how these viruses recognize different receptors. 19901337 8 19 theme virus-receptor 1178:1191 arg1 interactions 1193:1204 virus-receptor interactions 1178:1204 virus-receptor interactions 1178:1204 These results provide a structural basis for understanding viral evolution and virus-receptor interactions. 19901337 2 20 used used 303:306 arg2 ACE2 290:293 ACE2 290:293 ACE2 290:293 Incidentally, ACE2 is also used by group II SARS coronavirus (SARS-CoV). 19901337 4 21 theme crystal 522:528 arg1 structure 530:538 the crystal structure 518:538 the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2 518:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 1 22 theme only 182:185 arg1 coronavirus 114:124 NL63 coronavirus 109:124 NL63 coronavirus (NL63-CoV) 109:135 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 22 theme only 182:185 arg1 coronavirus 195:205 the only group I coronavirus 178:205 the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor 178:273 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 8 23 theme structural 1123:1132 arg1 basis 1134:1138 a structural basis 1121:1138 a structural basis for understanding viral evolution and virus-receptor interactions 1121:1204 These results provide a structural basis for understanding viral evolution and virus-receptor interactions. 19901337 4 24 theme NL63-CoV 543:550 arg1 RBD 591:593 RBD 591:593 RBD 591:593 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 4 24 theme NL63-CoV 543:550 arg1 domain 583:588 NL63-CoV spike protein receptor-binding domain 543:588 NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2 543:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 6 25 theme RBD 841:843 arg1 cores 845:849 RBD cores 841:849 RBD cores 841:849 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 1 26 theme respiratory 156:166 arg1 virus 168:172 a prevalent human respiratory virus 138:172 a prevalent human respiratory virus 138:172 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 26 theme respiratory 156:166 arg1 coronavirus 114:124 NL63 coronavirus 109:124 NL63 coronavirus (NL63-CoV) 109:135 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 27 theme group 187:191 arg1 coronavirus 114:124 NL63 coronavirus 109:124 NL63 coronavirus (NL63-CoV) 109:135 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 27 theme group 187:191 arg1 coronavirus 195:205 the only group I coronavirus 178:205 the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor 178:273 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 0 28 theme coronavirus 38:48 arg1 domain 67:72 NL63 respiratory coronavirus receptor-binding domain 21:72 NL63 respiratory coronavirus receptor-binding domain 21:72 Crystal structure of NL63 respiratory coronavirus receptor-binding domain complexed with its human receptor. 19901337 1 29 theme I 193:193 arg1 coronavirus 114:124 NL63 coronavirus 109:124 NL63 coronavirus (NL63-CoV) 109:135 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 29 theme I 193:193 arg1 coronavirus 195:205 the only group I coronavirus 178:205 the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor 178:273 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 0 30 theme respiratory 26:36 arg1 domain 67:72 NL63 respiratory coronavirus receptor-binding domain 21:72 NL63 respiratory coronavirus receptor-binding domain 21:72 Crystal structure of NL63 respiratory coronavirus receptor-binding domain complexed with its human receptor. 19901337 3 31 theme different 369:377 arg1 coronaviruses 389:401 coronaviruses 389:401 coronaviruses 389:401 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 3 31 theme different 369:377 arg1 groups 379:384 different groups 369:384 different groups of coronaviruses 369:401 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 6 32 theme virus-binding 931:943 arg1 " 952:952 a "virus-binding hotspot" 928:952 a "virus-binding hotspot" on ACE2 928:960 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 3 33 theme same 417:420 arg1 receptor 422:429 the same receptor 413:429 the same receptor 413:429 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 0 34 theme domain 67:72 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of NL63 respiratory coronavirus receptor-binding domain 0:72 Crystal structure of NL63 respiratory coronavirus receptor-binding domain complexed with its human receptor. 19901337 6 35 theme ACE2 895:898 arg1 regions 900:906 common ACE2 regions 888:906 common ACE2 regions 888:906 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 5 36 theme novel 642:646 arg1 structure 667:675 a novel beta-sandwich core structure 640:675 a novel beta-sandwich core structure consisting of 2 layers of beta-sheets 640:713 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 6 37 theme structural 818:827 arg1 homology 829:836 no structural homology 815:836 no structural homology 815:836 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 3 38 theme group 451:455 arg1 coronaviruses 459:471 homologous group I coronaviruses 440:471 homologous group I coronaviruses 440:471 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 0 39 theme receptor-binding 50:65 arg1 domain 67:72 NL63 respiratory coronavirus receptor-binding domain 21:72 NL63 respiratory coronavirus receptor-binding domain 21:72 Crystal structure of NL63 respiratory coronavirus receptor-binding domain complexed with its human receptor. 19901337 6 40 theme common 888:893 arg1 regions 900:906 common ACE2 regions 888:906 common ACE2 regions 888:906 NL63-CoV and SARS-CoV have no structural homology in RBD cores or RBMs; yet the 2 viruses recognize common ACE2 regions, largely because of a "virus-binding hotspot" on ACE2. 19901337 5 41 theme beta-sandwich 648:660 arg1 structure 667:675 a novel beta-sandwich core structure 640:675 a novel beta-sandwich core structure consisting of 2 layers of beta-sheets 640:713 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 3 42 theme I 457:457 arg1 coronaviruses 459:471 homologous group I coronaviruses 440:471 homologous group I coronaviruses 440:471 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 5 43 theme core 662:665 arg1 structure 667:675 a novel beta-sandwich core structure 640:675 a novel beta-sandwich core structure consisting of 2 layers of beta-sheets 640:713 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 7 44 theme group 969:973 arg1 coronaviruses 977:989 group I coronaviruses 969:989 group I coronaviruses 969:989 Among group I coronaviruses, RBD cores are conserved but RBMs are variable, explaining how these viruses recognize different receptors. 19901337 1 45 theme prevalent 140:148 arg1 virus 168:172 a prevalent human respiratory virus 138:172 a prevalent human respiratory virus 138:172 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 45 theme prevalent 140:148 arg1 coronavirus 114:124 NL63 coronavirus 109:124 NL63 coronavirus (NL63-CoV) 109:135 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 5 46 contain has 636:638 arg1 RBD 632:634 NL63-CoV RBD 623:634 NL63-CoV RBD 623:634 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 5 46 contain has 636:638 arg2 structure 667:675 a novel beta-sandwich core structure 640:675 a novel beta-sandwich core structure consisting of 2 layers of beta-sheets 640:713 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 7 47 theme RBD 992:994 arg1 cores 996:1000 RBD cores 992:1000 RBD cores 992:1000 Among group I coronaviruses, RBD cores are conserved but RBMs are variable, explaining how these viruses recognize different receptors. 19901337 4 48 theme human 611:615 arg1 ACE2 617:620 human ACE2 611:620 human ACE2 611:620 We determined the crystal structure of NL63-CoV spike protein receptor-binding domain (RBD) complexed with human ACE2. 19901337 3 49 theme different 483:491 arg1 receptors 493:501 different receptors 483:501 different receptors 483:501 We investigated how different groups of coronaviruses recognize the same receptor, whereas homologous group I coronaviruses recognize different receptors. 19901337 5 50 theme discontinuous 729:741 arg1 RBMs 768:771 RBMs 768:771 RBMs 768:771 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 5 50 theme discontinuous 729:741 arg1 motifs 760:765 3 discontinuous receptor-binding motifs 727:765 3 discontinuous receptor-binding motifs (RBMs) 727:772 NL63-CoV RBD has a novel beta-sandwich core structure consisting of 2 layers of beta-sheets, presenting 3 discontinuous receptor-binding motifs (RBMs) to bind ACE2. 19901337 2 51 theme SARS 320:323 arg1 SARS-CoV 338:345 SARS-CoV 338:345 SARS-CoV 338:345 Incidentally, ACE2 is also used by group II SARS coronavirus (SARS-CoV). 19901337 2 51 theme SARS 320:323 arg1 coronavirus 325:335 group II SARS coronavirus 311:335 group II SARS coronavirus (SARS-CoV) 311:346 Incidentally, ACE2 is also used by group II SARS coronavirus (SARS-CoV). 19901337 1 52 theme angiotensin-converting 220:241 arg1 enzyme 243:248 angiotensin-converting enzyme 2 220:250 angiotensin-converting enzyme 2 (ACE2) 220:257 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 19901337 1 52 theme angiotensin-converting 220:241 arg1 ACE2 253:256 ACE2 253:256 ACE2 253:256 NL63 coronavirus (NL63-CoV), a prevalent human respiratory virus, is the only group I coronavirus known to use angiotensin-converting enzyme 2 (ACE2) as its receptor. 22579623 0 0 theme quaternary 75:84 arg1 structure 86:94 TLR3ecd:3Fabs quaternary structure 61:94 TLR3ecd:3Fabs quaternary structure 61:94 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 3 1 theme formation 639:647 arg1 inhibition 622:631 inhibition 622:631 inhibition of SU formation 622:647 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 0 2 theme TLR3ecd:3Fabs 61:73 arg1 structure 86:94 TLR3ecd:3Fabs quaternary structure 61:94 TLR3ecd:3Fabs quaternary structure 61:94 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 6 3 theme single 1122:1127 arg1 SUs 1142:1144 single and multiple SUs 1122:1144 single and multiple SUs 1122:1144 This model is further supported by cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs. 22579623 4 4 theme SU 799:800 arg1 formation 802:810 minimal SU formation 791:810 minimal SU formation with short dsRNA 791:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 4 5 from sites 697:701 arg1 at 694:695 TLR3ecd at sites 686:701 TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA 686:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 4 5 from sites 697:701 arg1 TLR3ecd 686:692 TLR3ecd at sites 686:701 TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA 686:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 3 6 theme binding 485:491 arg1 site 493:496 the C-terminal dsRNA binding site 464:496 the C-terminal dsRNA binding site 464:496 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 5 7 theme ligand 1010:1015 arg1 length 990:995 the length 986:995 the length of the dsRNA ligand 986:1015 Molecular modeling based on the co-structure rationalizes these observations by showing that both Fab12 and Fab1068 prevent lateral clustering of SUs along the length of the dsRNA ligand. 22579623 4 8 theme minimal 791:797 arg1 formation 802:810 minimal SU formation 791:810 minimal SU formation with short dsRNA 791:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 4 9 theme TLR3ecd 686:692 arg1 sites 697:701 TLR3ecd at sites 686:701 TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA 686:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 6 10 theme dsRNA 1084:1088 arg1 ligands 1090:1096 dsRNA ligands 1084:1096 dsRNA ligands of lengths that support single and multiple SUs 1084:1144 This model is further supported by cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs. 22579623 3 11 theme C-terminal 468:477 arg1 site 493:496 the C-terminal dsRNA binding site 464:496 the C-terminal dsRNA binding site 464:496 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 1 12 theme immune 166:171 arg1 response 173:180 an innate immune response 156:180 an innate immune response 156:180 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 2 13 theme ectodomain 343:352 arg1 structure 319:327 the crystal structure 307:327 the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments 307:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 4 14 dep N- 721:722 arg1 the 717:719 the 717:719 the 717:719 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 3 15 theme dsRNA 479:483 arg1 site 493:496 the C-terminal dsRNA binding site 464:496 the C-terminal dsRNA binding site 464:496 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 1 16 theme innate 159:164 arg1 response 173:180 an innate immune response 156:180 an innate immune response 156:180 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 2 17 theme human 332:336 arg1 TLR3ecd 355:361 TLR3ecd 355:361 TLR3ecd 355:361 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 2 17 theme human 332:336 arg1 ectodomain 343:352 human TLR3 ectodomain 332:352 human TLR3 ectodomain (TLR3ecd) 332:362 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 0 18 theme Lateral 0:6 arg1 clustering 8:17 Lateral clustering 0:17 Lateral clustering of TLR3 0:25 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 2 19 from structure 319:327 arg1 complex 380:386 a quaternary complex 367:386 a quaternary complex with three neutralizing Fab fragments 367:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 3 20 theme biochemical 506:516 arg1 assays 518:523 biochemical assays 506:523 biochemical assays 506:523 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 2 21 theme quaternary 369:378 arg1 complex 380:386 a quaternary complex 367:386 a quaternary complex with three neutralizing Fab fragments 367:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 1 22 theme double-stranded 245:259 arg1 RNA 261:263 one double-stranded RNA 241:263 one double-stranded RNA (dsRNA) 241:271 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 1 22 theme double-stranded 245:259 arg1 dsRNA 266:270 dsRNA 266:270 dsRNA 266:270 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 6 23 theme multiple 1133:1140 arg1 SUs 1142:1144 single and multiple SUs 1122:1144 single and multiple SUs 1122:1144 This model is further supported by cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs. 22579623 1 24 theme Toll-like 97:105 arg1 TLR3 119:122 TLR3 119:122 TLR3 119:122 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 1 24 theme Toll-like 97:105 arg1 receptor 107:114 Toll-like receptor 3 97:116 Toll-like receptor 3 (TLR3) 97:123 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 4 25 theme distinct 703:710 arg1 sites 697:701 TLR3ecd at sites 686:701 TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA 686:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 6 26 theme lengths 1101:1107 arg1 ligands 1090:1096 dsRNA ligands 1084:1096 dsRNA ligands of lengths that support single and multiple SUs 1084:1144 This model is further supported by cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs. 22579623 6 27 theme cell-based 1053:1062 arg1 results 1070:1076 cell-based assay results 1053:1076 cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs 1053:1144 This model is further supported by cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs. 22579623 7 28 theme SUs 1225:1227 arg1 clustering 1211:1220 lateral clustering 1203:1220 lateral clustering of SUs 1203:1227 Thus, their antagonism of TLR3 signaling indicates that lateral clustering of SUs is required for TLR3 signal transduction. 22579623 0 29 theme dsRNA 27:31 arg1 units 43:47 dsRNA signaling units 27:47 dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure 27:94 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 0 30 dep units 43:47 arg1 clustering 8:17 Lateral clustering 0:17 Lateral clustering of TLR3 0:25 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 4 31 with formation 802:810 arg1 dsRNA 823:827 short dsRNA 817:827 short dsRNA 817:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 7 32 theme lateral 1203:1209 arg1 clustering 1211:1220 lateral clustering 1203:1220 lateral clustering of SUs 1203:1227 Thus, their antagonism of TLR3 signaling indicates that lateral clustering of SUs is required for TLR3 signal transduction. 22579623 2 33 theme Fab 412:414 arg1 fragments 416:424 three neutralizing Fab fragments 393:424 three neutralizing Fab fragments 393:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 0 34 theme TLR3 22:25 arg1 clustering 8:17 Lateral clustering 0:17 Lateral clustering of TLR3 0:25 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 2 35 theme neutralizing 399:410 arg1 fragments 416:424 three neutralizing Fab fragments 393:424 three neutralizing Fab fragments 393:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 5 36 theme dsRNA 1004:1008 arg1 ligand 1010:1015 the dsRNA ligand 1000:1015 the dsRNA ligand 1000:1015 Molecular modeling based on the co-structure rationalizes these observations by showing that both Fab12 and Fab1068 prevent lateral clustering of SUs along the length of the dsRNA ligand. 22579623 5 37 theme lateral 954:960 arg1 clustering 962:971 lateral clustering 954:971 lateral clustering of SUs 954:978 Molecular modeling based on the co-structure rationalizes these observations by showing that both Fab12 and Fab1068 prevent lateral clustering of SUs along the length of the dsRNA ligand. 22579623 0 38 theme signaling 33:41 arg1 units 43:47 dsRNA signaling units 27:47 dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure 27:94 Lateral clustering of TLR3:dsRNA signaling units revealed by TLR3ecd:3Fabs quaternary structure. 22579623 3 39 theme SU 636:637 arg1 formation 639:647 SU formation 636:647 SU formation 636:647 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 6 40 theme assay 1064:1068 arg1 results 1070:1076 cell-based assay results 1053:1076 cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs 1053:1144 This model is further supported by cell-based assay results using dsRNA ligands of lengths that support single and multiple SUs. 22579623 4 41 from regions 739:745 arg1 distinct 703:710 distinct 703:710 distinct 703:710 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 5 42 theme Molecular 830:838 arg1 modeling 840:847 Molecular modeling 830:847 Molecular modeling based on the co-structure 830:873 Molecular modeling based on the co-structure rationalizes these observations by showing that both Fab12 and Fab1068 prevent lateral clustering of SUs along the length of the dsRNA ligand. 22579623 4 43 theme C-terminal 728:737 arg1 regions 739:745 C-terminal regions 728:745 C-terminal regions 728:745 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 5 44 theme SUs 976:978 arg1 clustering 962:971 lateral clustering 954:971 lateral clustering of SUs 954:978 Molecular modeling based on the co-structure rationalizes these observations by showing that both Fab12 and Fab1068 prevent lateral clustering of SUs along the length of the dsRNA ligand. 22579623 3 45 theme TLR3ecd 552:558 arg1 interaction 537:547 the interaction 533:547 the interaction of TLR3ecd with dsRNA 533:569 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 2 46 with complex 380:386 arg1 fragments 416:424 three neutralizing Fab fragments 393:424 three neutralizing Fab fragments 393:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 7 47 theme TLR3 1173:1176 arg1 signaling 1178:1186 TLR3 signaling 1173:1186 TLR3 signaling 1173:1186 Thus, their antagonism of TLR3 signaling indicates that lateral clustering of SUs is required for TLR3 signal transduction. 22579623 3 48 dep overlaps 455:462 arg1 blocks 526:531 blocks 526:531 blocks 526:531 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 4 49 theme short 817:821 arg1 dsRNA 823:827 short dsRNA 817:827 short dsRNA 817:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 4 50 theme at 694:695 arg1 sites 697:701 TLR3ecd at sites 686:701 TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA 686:827 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 7 51 theme signaling 1178:1186 arg1 antagonism 1159:1168 their antagonism 1153:1168 their antagonism of TLR3 signaling 1153:1186 Thus, their antagonism of TLR3 signaling indicates that lateral clustering of SUs is required for TLR3 signal transduction. 22579623 7 52 theme TLR3 1245:1248 arg1 transduction 1257:1268 TLR3 signal transduction 1245:1268 TLR3 signal transduction 1245:1268 Thus, their antagonism of TLR3 signaling indicates that lateral clustering of SUs is required for TLR3 signal transduction. 22579623 1 53 theme signaling 209:217 arg1 unit 219:222 a signaling unit 207:222 a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules 207:294 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 1 53 theme signaling 209:217 arg1 SU 225:226 SU 225:226 SU 225:226 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 3 54 with interaction 537:547 arg1 dsRNA 565:569 dsRNA 565:569 dsRNA 565:569 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 3 55 theme TLR3 599:602 arg1 signaling 604:612 TLR3 signaling 599:612 TLR3 signaling 599:612 Fab15 binds an epitope that overlaps the C-terminal dsRNA binding site and, in biochemical assays, blocks the interaction of TLR3ecd with dsRNA, thus directly antagonizing TLR3 signaling through inhibition of SU formation. 22579623 7 56 theme signal 1250:1255 arg1 transduction 1257:1268 TLR3 signal transduction 1245:1268 TLR3 signal transduction 1245:1268 Thus, their antagonism of TLR3 signaling indicates that lateral clustering of SUs is required for TLR3 signal transduction. 22579623 4 57 from N- 721:722 arg1 distinct 703:710 distinct 703:710 distinct 703:710 In contrast, Fab12 and Fab1068 bind TLR3ecd at sites distinct from the N- and C-terminal regions that interact with dsRNA and do not inhibit minimal SU formation with short dsRNA. 22579623 2 58 theme TLR3 338:341 arg1 TLR3ecd 355:361 TLR3ecd 355:361 TLR3ecd 355:361 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 2 58 theme TLR3 338:341 arg1 ectodomain 343:352 human TLR3 ectodomain 332:352 human TLR3 ectodomain (TLR3ecd) 332:362 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 22579623 1 59 theme unit 219:222 arg1 formation 194:202 the formation 190:202 the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules 190:294 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 1 60 theme TLR3 281:284 arg1 molecules 286:294 two TLR3 molecules 277:294 two TLR3 molecules 277:294 Toll-like receptor 3 (TLR3) recognizes dsRNA and initiates an innate immune response through the formation of a signaling unit (SU) composed of one double-stranded RNA (dsRNA) and two TLR3 molecules. 22579623 2 61 theme crystal 311:317 arg1 structure 319:327 the crystal structure 307:327 the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments 307:424 We report the crystal structure of human TLR3 ectodomain (TLR3ecd) in a quaternary complex with three neutralizing Fab fragments. 16638743 5 0 theme FGF23 849:853 arg1 model 840:844 a novel posttranslational regulatory model 803:844 a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23 803:937 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 16638743 4 1 theme fibroblast 562:571 arg1 factor 580:585 the phosphaturic factor fibroblast growth factor 23 538:588 the phosphaturic factor fibroblast growth factor 23 (FGF23) 538:596 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 1 theme fibroblast 562:571 arg1 FGF23 591:595 FGF23 591:595 FGF23 591:595 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 2 theme factor 555:560 arg1 factor 580:585 the phosphaturic factor fibroblast growth factor 23 538:588 the phosphaturic factor fibroblast growth factor 23 (FGF23) 538:596 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 2 theme factor 555:560 arg1 FGF23 591:595 FGF23 591:595 FGF23 591:595 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 2 3 theme rare 307:310 arg1 disorder 342:349 the rare autosomal recessive metabolic disorder 303:349 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 4 theme disorder 342:349 arg1 calcinosis 368:377 the rare autosomal recessive metabolic disorder familial tumoral calcinosis 303:377 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 4 theme disorder 342:349 arg1 OMIM 380:383 OMIM 211900 380:390 OMIM 211900 380:390 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 5 theme glycosyltransferase 169:187 arg1 GalNAc-T3 201:209 the glycosyltransferase polypeptide GalNAc-T3 165:209 the glycosyltransferase polypeptide GalNAc-T3 165:209 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 4 6 gly O-glycosylation 663:677 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 5 7 theme novel 805:809 arg1 model 840:844 a novel posttranslational regulatory model 803:844 a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23 803:937 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 16638743 1 8 theme fibroblast 80:89 arg1 factor 98:103 fibroblast growth factor 23 80:106 fibroblast growth factor 23 80:106 Secretion of fibroblast growth factor 23 requires O-glycosylation. 16638743 4 9 theme sequence 734:741 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 5 10 theme posttranslational 811:827 arg1 model 840:844 a novel posttranslational regulatory model 803:844 a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23 803:937 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 16638743 1 11 theme growth 91:96 arg1 factor 98:103 fibroblast growth factor 23 80:106 fibroblast growth factor 23 80:106 Secretion of fibroblast growth factor 23 requires O-glycosylation. 16638743 4 12 theme recognition 722:732 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 2 13 from Mutations 134:142 arg1 gene 151:154 the gene 147:154 the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, 147:262 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 1 14 theme factor 98:103 arg1 Secretion 67:75 Secretion 67:75 Secretion of fibroblast growth factor 23 67:106 Secretion of fibroblast growth factor 23 requires O-glycosylation. 16638743 3 15 theme massive 463:469 arg1 calcifications 479:492 massive ectopic calcifications 463:492 massive ectopic calcifications 463:492 Familial tumoral calcinosis is associated with hyperphosphatemia and massive ectopic calcifications. 16638743 0 16 theme Polypeptide 0:10 arg1 GalNAc-transferase 12:29 Polypeptide GalNAc-transferase 0:29 Polypeptide GalNAc-transferase 0:29 Polypeptide GalNAc-transferase T3 and familial tumoral calcinosis. 16638743 2 17 theme metabolic 332:340 arg1 disorder 342:349 the rare autosomal recessive metabolic disorder 303:349 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 3 18 theme ectopic 471:477 arg1 calcifications 479:492 massive ectopic calcifications 463:492 massive ectopic calcifications 463:492 Familial tumoral calcinosis is associated with hyperphosphatemia and massive ectopic calcifications. 16638743 3 19 theme Familial 394:401 arg1 calcinosis 411:420 Familial tumoral calcinosis 394:420 Familial tumoral calcinosis 394:420 Familial tumoral calcinosis is associated with hyperphosphatemia and massive ectopic calcifications. 16638743 2 20 theme recessive 322:330 arg1 disorder 342:349 the rare autosomal recessive metabolic disorder 303:349 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 4 21 theme subtilisin-like 684:698 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 3 22 theme tumoral 403:409 arg1 calcinosis 411:420 Familial tumoral calcinosis 394:420 Familial tumoral calcinosis 394:420 Familial tumoral calcinosis is associated with hyperphosphatemia and massive ectopic calcifications. 16638743 2 23 theme autosomal 312:320 arg1 disorder 342:349 the rare autosomal recessive metabolic disorder 303:349 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 0 24 theme tumoral 47:53 arg1 calcinosis 55:64 familial tumoral calcinosis 38:64 familial tumoral calcinosis 38:64 Polypeptide GalNAc-transferase T3 and familial tumoral calcinosis. 16638743 4 25 theme FGF23 777:781 arg1 processing 763:772 processing 763:772 processing of FGF23 763:781 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 26 theme convertase 711:720 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 0 27 theme familial 38:45 arg1 calcinosis 55:64 familial tumoral calcinosis 38:64 familial tumoral calcinosis 38:64 Polypeptide GalNAc-transferase T3 and familial tumoral calcinosis. 16638743 4 28 theme factor 580:585 arg1 secretion 525:533 the secretion 521:533 the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) 521:596 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 5 29 theme competing 865:873 arg1 O-glycosylation 875:889 competing O-glycosylation 865:889 competing O-glycosylation 865:889 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 16638743 4 30 theme proprotein 700:709 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 31 theme growth 573:578 arg1 factor 580:585 the phosphaturic factor fibroblast growth factor 23 538:588 the phosphaturic factor fibroblast growth factor 23 (FGF23) 538:596 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 31 theme growth 573:578 arg1 FGF23 591:595 FGF23 591:595 FGF23 591:595 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 2 32 theme polypeptide 189:199 arg1 GalNAc-T3 201:209 the glycosyltransferase polypeptide GalNAc-T3 165:209 the glycosyltransferase polypeptide GalNAc-T3 165:209 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 33 theme O-glycosylation 247:261 arg1 initiation 233:242 initiation 233:242 initiation of O-glycosylation 233:261 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 34 theme calcinosis 368:377 arg1 cause 294:298 a cause 292:298 a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 292:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 34 theme calcinosis 368:377 arg1 Mutations 134:142 Mutations 134:142 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, 134:262 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 35 theme tumoral 360:366 arg1 calcinosis 368:377 the rare autosomal recessive metabolic disorder familial tumoral calcinosis 303:377 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 35 theme tumoral 360:366 arg1 OMIM 380:383 OMIM 211900 380:390 OMIM 211900 380:390 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 0 36 dep T3 31:32 arg1 GalNAc-transferase 12:29 Polypeptide GalNAc-transferase 0:29 Polypeptide GalNAc-transferase 0:29 Polypeptide GalNAc-transferase T3 and familial tumoral calcinosis. 16638743 2 37 theme familial 351:358 arg1 calcinosis 368:377 the rare autosomal recessive metabolic disorder familial tumoral calcinosis 303:377 the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900) 303:391 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 2 37 theme familial 351:358 arg1 OMIM 380:383 OMIM 211900 380:390 OMIM 211900 380:390 Mutations in the gene encoding the glycosyltransferase polypeptide GalNAc-T3, which is involved in initiation of O-glycosylation, were recently identified as a cause of the rare autosomal recessive metabolic disorder familial tumoral calcinosis (OMIM 211900). 16638743 5 38 theme regulatory 829:838 arg1 model 840:844 a novel posttranslational regulatory model 803:844 a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23 803:937 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 16638743 5 39 theme intact 926:931 arg1 FGF23 933:937 intact FGF23 926:937 intact FGF23 926:937 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 16638743 4 40 from O-glycosylation 663:677 arg1 motif 743:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 a subtilisin-like proprotein convertase recognition sequence motif 682:747 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 41 theme phosphaturic 542:553 arg1 factor 580:585 the phosphaturic factor fibroblast growth factor 23 538:588 the phosphaturic factor fibroblast growth factor 23 (FGF23) 538:596 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 4 41 theme phosphaturic 542:553 arg1 FGF23 591:595 FGF23 591:595 FGF23 591:595 Here, we demonstrate that the secretion of the phosphaturic factor fibroblast growth factor 23 (FGF23) requires O-glycosylation, and that GalNAc-T3 selectively directs O-glycosylation in a subtilisin-like proprotein convertase recognition sequence motif, which blocks processing of FGF23. 16638743 5 42 theme protease 895:902 arg1 processing 904:913 protease processing 895:913 protease processing 895:913 The study suggests a novel posttranslational regulatory model of FGF23 involving competing O-glycosylation and protease processing to produce intact FGF23. 10074346 4 0 theme amino 772:776 arg1 Aspbeta398 800:809 Aspbeta398 800:809 Aspbeta398 800:809 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 0 theme amino 772:776 arg1 Glubeta397 785:794 Glubeta397 785:794 Glubeta397 785:794 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 0 theme amino 772:776 arg1 acids 778:782 two negatively charged amino acids 749:782 two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 749:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 5 1 theme GPRPam 988:993 arg1 absence 958:964 the absence 954:964 the absence of the peptide ligand GPRPam 954:993 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 0 2 from changes 15:21 arg1 fragments 26:34 fragments D and double-D 26:49 fragments D and double-D from human fibrin(ogen) 26:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 2 from changes 15:21 arg1 double-D 42:49 double-D 42:49 double-D 42:49 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 2 from changes 15:21 arg1 fibrin 62:67 human fibrin 56:67 human fibrin(ogen) 56:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 2 from changes 15:21 arg1 D 36:36 D 36:36 D 36:36 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 2 from changes 15:21 arg1 ogen 69:72 ogen 69:72 ogen 69:72 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 4 3 theme charged 764:770 arg1 Aspbeta398 800:809 Aspbeta398 800:809 Aspbeta398 800:809 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 3 theme charged 764:770 arg1 Glubeta397 785:794 Glubeta397 785:794 Glubeta397 785:794 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 3 theme charged 764:770 arg1 acids 778:782 two negatively charged amino acids 749:782 two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 749:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 1 4 theme fragment 147:154 arg1 double-D 156:163 fragment double-D 147:163 fragment double-D from human fibrin 147:181 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 5 theme peptide 234:240 arg1 ligands 242:248 the peptide ligands 230:248 the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively 230:340 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 2 6 dep fragments 459:467 arg1 fragments 459:467 fragments D and double-D 459:482 fragments D and double-D with the peptide GHRPam alone 459:512 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 6 dep fragments 459:467 arg1 double-D 475:482 double-D 475:482 double-D 475:482 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 6 dep fragments 459:467 arg1 D 469:469 D 469:469 D 469:469 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 4 7 theme acids 778:782 arg1 " 744:744 a moveable "flap" 728:744 a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 728:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 2 8 theme ligand 557:562 arg1 absence 534:540 the absence 530:540 the absence of any peptide ligand 530:562 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 1 9 theme ligands 242:248 arg1 absence 219:225 absence 219:225 absence 219:225 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 9 theme ligands 242:248 arg1 presence 206:213 presence 206:213 presence 206:213 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 5 10 theme peptide 973:979 arg1 GPRPam 988:993 the peptide ligand GPRPam 969:993 the peptide ligand GPRPam 969:993 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 2 11 dep determined 386:395 arg1 double-D 518:525 double-D 518:525 double-D in the absence of any peptide ligand 518:562 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 11 dep determined 386:395 arg1 fragments 459:467 fragments D and double-D 459:482 fragments D and double-D with the peptide GHRPam alone 459:512 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 11 dep determined 386:395 arg1 double-D 475:482 double-D 475:482 double-D 475:482 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 11 dep determined 386:395 arg1 D 469:469 D 469:469 D 469:469 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 1 12 from structure 134:142 arg1 fibrin 176:181 human fibrin 170:181 human fibrin 170:181 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 13 theme double-D 156:163 arg1 structure 134:142 The structure 130:142 The structure of fragment double-D from human fibrin 130:181 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 3 14 theme conformational 619:632 arg1 changes 634:640 conformational changes 619:640 conformational changes that are brought about by the various knob-hole interactions 619:701 Comparison of the structures has revealed a series of conformational changes that are brought about by the various knob-hole interactions. 10074346 0 15 theme Conformational 0:13 arg1 changes 15:21 Conformational changes 0:21 Conformational changes in fragments D and double-D from human fibrin(ogen) 0:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 3 16 theme changes 634:640 arg1 series 609:614 a series 607:614 a series of conformational changes that are brought about by the various knob-hole interactions 607:701 Comparison of the structures has revealed a series of conformational changes that are brought about by the various knob-hole interactions. 10074346 2 17 with double-D 518:525 arg1 GHRPam 501:506 the peptide GHRPam 489:506 the peptide GHRPam alone 489:512 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 1 18 theme human 170:174 arg1 fibrin 176:181 human fibrin 170:181 human fibrin 170:181 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 19 theme fibrinopeptides 304:318 arg1 removal 293:299 the removal 289:299 the removal of fibrinopeptides A and B 289:326 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 20 dep fibrinopeptides 304:318 arg1 fibrinopeptides 304:318 fibrinopeptides A and B 304:326 fibrinopeptides A and B 304:326 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 20 dep fibrinopeptides 304:318 arg1 B 326:326 B 326:326 B 326:326 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 20 dep fibrinopeptides 304:318 arg1 A 320:320 A 320:320 A 320:320 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 4 21 theme beta-chain 918:927 arg1 pocket 929:934 the beta-chain pocket 914:934 the beta-chain pocket 914:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 5 22 theme new 1038:1040 arg1 pocket 1028:1033 the gamma-chain pocket 1012:1033 the gamma-chain pocket 1012:1033 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 5 22 theme new 1038:1040 arg1 site 1058:1061 a new calcium-binding site 1036:1061 a new calcium-binding site being formed concomitantly 1036:1088 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 0 23 theme ligand 100:105 arg1 Gly-His-Arg-Pro-amide 107:127 the peptide ligand Gly-His-Arg-Pro-amide 88:127 the peptide ligand Gly-His-Arg-Pro-amide 88:127 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 2 24 theme peptide 549:555 arg1 ligand 557:562 any peptide ligand 545:562 any peptide ligand 545:562 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 5 25 theme calcium-binding 1042:1056 arg1 pocket 1028:1033 the gamma-chain pocket 1012:1033 the gamma-chain pocket 1012:1033 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 5 25 theme calcium-binding 1042:1056 arg1 site 1058:1061 a new calcium-binding site 1036:1061 a new calcium-binding site being formed concomitantly 1036:1088 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 0 26 theme peptide 92:98 arg1 Gly-His-Arg-Pro-amide 107:127 the peptide ligand Gly-His-Arg-Pro-amide 88:127 the peptide ligand Gly-His-Arg-Pro-amide 88:127 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 4 27 theme atom 880:883 arg1 bridge 885:890 a calcium atom bridge 870:890 a calcium atom bridge 870:890 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 2 28 from double-D 518:525 arg1 absence 534:540 the absence 530:540 the absence of any peptide ligand 530:562 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 4 29 theme moveable 730:737 arg1 " 744:744 a moveable "flap" 728:744 a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 728:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 2 30 theme crystal 357:363 arg1 structures 365:374 six crystal structures 353:374 six crystal structures 353:374 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 30 theme crystal 357:363 arg1 All 343:345 All 343:345 All 343:345 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 4 31 theme calcium 872:878 arg1 bridge 885:890 a calcium atom bridge 870:890 a calcium atom bridge 870:890 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 32 dep acids 778:782 arg1 Aspbeta398 800:809 Aspbeta398 800:809 Aspbeta398 800:809 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 32 dep acids 778:782 arg1 Glubeta397 785:794 Glubeta397 785:794 Glubeta397 785:794 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 32 dep acids 778:782 arg1 acids 778:782 two negatively charged amino acids 749:782 two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 749:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 0 33 from fibrin 62:67 arg1 fragments 26:34 fragments D and double-D 26:49 fragments D and double-D from human fibrin(ogen) 26:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 33 from fibrin 62:67 arg1 double-D 42:49 double-D 42:49 double-D 42:49 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 33 from fibrin 62:67 arg1 changes 15:21 Conformational changes 0:21 Conformational changes in fragments D and double-D from human fibrin(ogen) 0:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 33 from fibrin 62:67 arg1 D 36:36 D 36:36 D 36:36 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 2 34 theme which 407:411 arg1 which 407:411 which 407:411 which 407:411 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 34 theme which 407:411 arg1 three 398:402 three 398:402 three 398:402 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 5 35 theme gamma-chain 1016:1026 arg1 pocket 1028:1033 the gamma-chain pocket 1012:1033 the gamma-chain pocket 1012:1033 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 5 35 theme gamma-chain 1016:1026 arg1 site 1058:1061 a new calcium-binding site 1036:1061 a new calcium-binding site being formed concomitantly 1036:1088 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 1 36 dep presence 206:213 arg1 the 202:204 the 202:204 the 202:204 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 4 37 theme flap 740:743 arg1 " 744:744 a moveable "flap" 728:744 a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 728:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 5 38 theme ligand 981:986 arg1 GPRPam 988:993 the peptide ligand GPRPam 969:993 the peptide ligand GPRPam 969:993 Additionally, in the absence of the peptide ligand GPRPam, GHRPam binds to the gamma-chain pocket, a new calcium-binding site being formed concomitantly. 10074346 3 39 theme various 672:678 arg1 interactions 690:701 the various knob-hole interactions 668:701 the various knob-hole interactions 668:701 Comparison of the structures has revealed a series of conformational changes that are brought about by the various knob-hole interactions. 10074346 2 40 theme peptide 493:499 arg1 GHRPam 501:506 the peptide GHRPam 489:506 the peptide GHRPam alone 489:512 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 3 41 theme structures 583:592 arg1 Comparison 565:574 Comparison 565:574 Comparison of the structures 565:592 Comparison of the structures has revealed a series of conformational changes that are brought about by the various knob-hole interactions. 10074346 4 42 theme coiled 853:858 arg1 coil 860:863 the coiled coil 849:863 the coiled coil with a calcium atom bridge 849:890 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 4 43 with coil 860:863 arg1 bridge 885:890 a calcium atom bridge 870:890 a calcium atom bridge 870:890 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 3 44 theme knob-hole 680:688 arg1 interactions 690:701 the various knob-hole interactions 668:701 the various knob-hole interactions 668:701 Comparison of the structures has revealed a series of conformational changes that are brought about by the various knob-hole interactions. 10074346 4 45 theme greatest 707:714 arg1 interest 716:723 greatest interest 707:723 greatest interest 707:723 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 0 46 dep fragments 26:34 arg1 fragments 26:34 fragments D and double-D 26:49 fragments D and double-D from human fibrin(ogen) 26:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 46 dep fragments 26:34 arg1 double-D 42:49 double-D 42:49 double-D 42:49 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 46 dep fragments 26:34 arg1 D 36:36 D 36:36 D 36:36 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 2 47 with fragments 459:467 arg1 GHRPam 501:506 the peptide GHRPam 489:506 the peptide GHRPam alone 489:512 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 48 from fragments 459:467 arg1 absence 534:540 the absence 530:540 the absence of any peptide ligand 530:562 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 2 49 theme first 439:443 arg1 time 445:448 the first time 435:448 the first time 435:448 All told, six crystal structures have been determined, three of which are reported here for the first time: namely, fragments D and double-D with the peptide GHRPam alone and double-D in the absence of any peptide ligand. 10074346 4 50 theme side 818:821 arg1 chains 823:828 side chains 818:828 side chains 818:828 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 10074346 1 51 from fibrin 176:181 arg1 structure 134:142 The structure 130:142 The structure of fragment double-D from human fibrin 130:181 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 1 51 from fibrin 176:181 arg1 double-D 156:163 fragment double-D 147:163 fragment double-D from human fibrin 147:181 The structure of fragment double-D from human fibrin has been solved in the presence and absence of the peptide ligands that simulate the two knobs exposed by the removal of fibrinopeptides A and B, respectively. 10074346 0 52 theme human 56:60 arg1 ogen 69:72 ogen 69:72 ogen 69:72 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 0 52 theme human 56:60 arg1 fibrin 62:67 human fibrin 56:67 human fibrin(ogen) 56:73 Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. 10074346 4 53 theme interest 716:723 arg1 " 744:744 a moveable "flap" 728:744 a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket 728:934 Of greatest interest is a moveable "flap" of two negatively charged amino acids (Glubeta397 and Aspbeta398) whose side chains are pinned back to the coiled coil with a calcium atom bridge until GHRPam occupies the beta-chain pocket. 9628725 0 0 theme bound 76:80 arg1 ligands 82:88 two different bound ligands 62:88 two different bound ligands 62:88 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 1 1 theme human 143:147 arg1 fibrin 149:154 human fibrin 143:154 human fibrin 143:154 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 2 theme X-ray 230:234 arg1 structure 236:244 the X-ray structure 226:244 the X-ray structure determined at 2.3 A 226:264 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 0 3 theme different 66:74 arg1 ligands 82:88 two different bound ligands 62:88 two different bound ligands 62:88 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 3 4 theme first 599:603 arg1 time 605:608 the first time 595:608 the first time 595:608 The structure shows for the first time that the beta-chain knob does indeed bind to a homologous hole on the beta-chain. 9628725 2 5 theme beta-chain 553:562 arg1 holes 564:568 the homologous beta-chain holes 538:568 the homologous beta-chain holes 538:568 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 1 6 from fibrin 149:154 arg1 double-D 128:135 double-D 128:135 double-D 128:135 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 6 from fibrin 149:154 arg1 D 125:125 Factor XIII-cross-linked fragment D 91:125 Factor XIII-cross-linked fragment D (double-D) from human fibrin 91:154 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 4 7 theme beta-chain 707:716 arg1 similar 746:752 similar 746:752 similar 746:752 The gamma- and beta-chain holes are structurally very similar, and it is remarkable they are able to distinguish between these two peptides that differ by a single amino acid. 9628725 4 7 theme beta-chain 707:716 arg1 holes 718:722 The gamma- and beta-chain holes 692:722 The gamma- and beta-chain holes 692:722 The gamma- and beta-chain holes are structurally very similar, and it is remarkable they are able to distinguish between these two peptides that differ by a single amino acid. 9628725 2 8 theme peptide 271:277 arg1 analogue 314:321 an analogue 311:321 an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A 311:395 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 8 theme peptide 271:277 arg1 Gly-Pro-Arg-Pro-amide 279:299 The peptide Gly-Pro-Arg-Pro-amide 267:299 The peptide Gly-Pro-Arg-Pro-amide 267:299 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 0 9 from fibrin 50:55 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 0:89 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 2 10 theme homologous 542:551 arg1 holes 564:568 the homologous beta-chain holes 538:568 the homologous beta-chain holes 538:568 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 11 theme knob 330:333 arg1 analogue 314:321 an analogue 311:321 an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A 311:395 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 11 theme knob 330:333 arg1 Gly-Pro-Arg-Pro-amide 279:299 The peptide Gly-Pro-Arg-Pro-amide 267:299 The peptide Gly-Pro-Arg-Pro-amide 267:299 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 3 12 theme homologous 657:666 arg1 hole 668:671 a homologous hole 655:671 a homologous hole on the beta-chain 655:689 The structure shows for the first time that the beta-chain knob does indeed bind to a homologous hole on the beta-chain. 9628725 3 13 from hole 668:671 arg1 beta-chain 680:689 the beta-chain 676:689 the beta-chain 676:689 The structure shows for the first time that the beta-chain knob does indeed bind to a homologous hole on the beta-chain. 9628725 2 14 located found 529:533 arg1 holes 564:568 the homologous beta-chain holes 538:568 the homologous beta-chain holes 538:568 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 14 located found 529:533 arg2 Gly-His-Arg-Pro-amide 460:480 the peptide Gly-His-Arg-Pro-amide 448:480 the peptide Gly-His-Arg-Pro-amide 448:480 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 15 theme thrombin-catalyzed 350:367 arg1 removal 369:375 the thrombin-catalyzed removal 346:375 the thrombin-catalyzed removal of fibrinopeptide A 346:395 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 0 16 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 0:89 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 3 17 theme beta-chain 619:628 arg1 knob 630:633 the beta-chain knob 615:633 the beta-chain knob 615:633 The structure shows for the first time that the beta-chain knob does indeed bind to a homologous hole on the beta-chain. 9628725 0 18 theme fragment 21:28 arg1 double-D 30:37 fragment double-D 21:37 fragment double-D 21:37 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 4 19 theme single 849:854 arg1 acid 862:865 a single amino acid 847:865 a single amino acid 847:865 The gamma- and beta-chain holes are structurally very similar, and it is remarkable they are able to distinguish between these two peptides that differ by a single amino acid. 9628725 1 20 theme Factor 91:96 arg1 double-D 128:135 double-D 128:135 double-D 128:135 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 20 theme Factor 91:96 arg1 D 125:125 Factor XIII-cross-linked fragment D 91:125 Factor XIII-cross-linked fragment D (double-D) from human fibrin 91:154 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 21 theme XIII-cross-linked 98:114 arg1 double-D 128:135 double-D 128:135 double-D 128:135 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 21 theme XIII-cross-linked 98:114 arg1 D 125:125 Factor XIII-cross-linked fragment D 91:125 Factor XIII-cross-linked fragment D (double-D) from human fibrin 91:154 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 2 22 theme beta-chain 508:517 arg1 knob 519:522 the beta-chain knob 504:522 the beta-chain knob 504:522 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 23 theme A 395:395 arg1 removal 369:375 the thrombin-catalyzed removal 346:375 the thrombin-catalyzed removal of fibrinopeptide A 346:395 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 0 24 theme double-D 30:37 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 0:89 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 4 25 theme amino 856:860 arg1 acid 862:865 a single amino acid 847:865 a single amino acid 847:865 The gamma- and beta-chain holes are structurally very similar, and it is remarkable they are able to distinguish between these two peptides that differ by a single amino acid. 9628725 5 26 theme gamma-chain 933:943 arg1 counterpart 945:955 its gamma-chain counterpart 929:955 its gamma-chain counterpart 929:955 Additionally, we have found that the beta-chain domain, like its gamma-chain counterpart, binds calcium. 9628725 1 27 theme different 196:204 arg1 ligands 214:220 two different peptide ligands 192:220 two different peptide ligands 192:220 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 2 28 theme gamma-chain 425:435 arg1 holes 437:441 the gamma-chain holes 421:441 the gamma-chain holes 421:441 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 2 29 theme peptide 452:458 arg1 Gly-His-Arg-Pro-amide 460:480 the peptide Gly-His-Arg-Pro-amide 448:480 the peptide Gly-His-Arg-Pro-amide 448:480 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 1 30 theme fragment 116:123 arg1 double-D 128:135 double-D 128:135 double-D 128:135 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 30 theme fragment 116:123 arg1 D 125:125 Factor XIII-cross-linked fragment D 91:125 Factor XIII-cross-linked fragment D (double-D) from human fibrin 91:154 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 0 31 theme human 44:48 arg1 fibrin 50:55 human fibrin 44:55 human fibrin with two different bound ligands 44:88 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 5 32 theme beta-chain 905:914 arg1 domain 916:921 the beta-chain domain 901:921 the beta-chain domain 901:921 Additionally, we have found that the beta-chain domain, like its gamma-chain counterpart, binds calcium. 9628725 4 33 theme gamma- 696:701 arg1 similar 746:752 similar 746:752 similar 746:752 The gamma- and beta-chain holes are structurally very similar, and it is remarkable they are able to distinguish between these two peptides that differ by a single amino acid. 9628725 4 33 theme gamma- 696:701 arg1 holes 718:722 The gamma- and beta-chain holes 692:722 The gamma- and beta-chain holes 692:722 The gamma- and beta-chain holes are structurally very similar, and it is remarkable they are able to distinguish between these two peptides that differ by a single amino acid. 9628725 1 34 theme peptide 206:212 arg1 ligands 214:220 two different peptide ligands 192:220 two different peptide ligands 192:220 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 0 35 with fibrin 50:55 arg1 ligands 82:88 two different bound ligands 62:88 two different bound ligands 62:88 Crystal structure of fragment double-D from human fibrin with two different bound ligands. 9628725 1 36 theme ligands 214:220 arg1 structure 236:244 the X-ray structure 226:244 the X-ray structure determined at 2.3 A 226:264 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 36 theme ligands 214:220 arg1 presence 180:187 the presence 176:187 the presence of two different peptide ligands 176:220 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 2 37 theme fibrinopeptide 380:393 arg1 A 395:395 fibrinopeptide A 380:395 fibrinopeptide A 380:395 The peptide Gly-Pro-Arg-Pro-amide, which is an analogue of the knob exposed by the thrombin-catalyzed removal of fibrinopeptide A, was found to reside in the gamma-chain holes, and the peptide Gly-His-Arg-Pro-amide, which corresponds to the beta-chain knob, was found in the homologous beta-chain holes. 9628725 1 38 link XIII-cross-linked 98:114 arg1 double-D 128:135 double-D 128:135 double-D 128:135 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 9628725 1 38 link XIII-cross-linked 98:114 arg1 D 125:125 Factor XIII-cross-linked fragment D 91:125 Factor XIII-cross-linked fragment D (double-D) from human fibrin 91:154 Factor XIII-cross-linked fragment D (double-D) from human fibrin was crystallized in the presence of two different peptide ligands and the X-ray structure determined at 2.3 A. 26859324 5 0 theme mu 765:766 arg1 COX-2 769:773 S530T murine (mu) COX-2 751:773 S530T murine (mu) COX-2 751:773 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 6 1 theme muCOX-2 1158:1164 arg1 structure 1174:1182 the S530T muCOX-2 crystal structure 1148:1182 the S530T muCOX-2 crystal structure 1148:1182 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 8 2 from acetylation 1467:1477 arg1 nanodiscs 1530:1538 nanodiscs 1530:1538 nanodiscs 1530:1538 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 8 2 from acetylation 1467:1477 arg1 systems 1518:1524 various detergent systems 1500:1524 various detergent systems 1500:1524 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 4 3 theme prostaglandin 688:700 arg1 product 702:708 the prostaglandin product 684:708 the prostaglandin product 684:708 Ser-530 has also been shown to influence the stereochemistry for the addition of oxygen to the prostaglandin product. 26859324 1 4 theme prostaglandins 267:280 arg1 formation 254:262 the formation 250:262 the formation of prostaglandins 250:280 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 5 5 theme crystal 729:735 arg1 structures 737:746 the crystal structures 725:746 the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate 725:850 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 7 6 theme mutant 1338:1343 arg1 analysis 1303:1310 functional analysis 1292:1310 functional analysis of the S530T/G533V double mutant 1292:1343 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 7 7 theme functional 1292:1301 arg1 analysis 1303:1310 functional analysis 1292:1310 functional analysis of the S530T/G533V double mutant 1292:1343 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 0 8 theme Products 94:101 arg1 Formation 81:89 the Formation 77:89 the Formation of Products with Reversed Stereochemistry 77:131 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 6 9 dep blocks 955:960 arg1 1 918:918 1 918:918 1 918:918 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 8 10 theme enzyme 1627:1632 arg1 domain 1613:1618 the membrane-binding domain 1592:1618 the membrane-binding domain of the enzyme 1592:1632 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 8 10 theme enzyme 1627:1632 arg1 enzyme 1627:1632 the enzyme 1623:1632 the enzyme 1623:1632 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 6 11 dep impede 1193:1198 arg1 3 1113:1113 3 1113:1113 3 1113:1113 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 7 12 theme working 1359:1365 arg1 hypothesis 1367:1376 a working hypothesis 1357:1376 a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2 1357:1435 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 6 13 theme crystal 1166:1172 arg1 structure 1174:1182 the S530T muCOX-2 crystal structure 1148:1182 the S530T muCOX-2 crystal structure 1148:1182 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 3 14 theme COX-2 484:488 arg1 acetylation 469:479 acetylation 469:479 acetylation of COX-2 469:488 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 7 15 theme S530T/G533V 1319:1329 arg1 mutant 1338:1343 the S530T/G533V double mutant 1315:1343 the S530T/G533V double mutant 1315:1343 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 3 16 theme activity 453:460 arg1 loss 445:448 complete loss 436:448 complete loss of activity 436:460 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 0 17 theme Reversed 108:115 arg1 Stereochemistry 117:131 Reversed Stereochemistry 108:131 Reversed Stereochemistry 108:131 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 5 18 theme COX-2 769:773 arg1 structures 737:746 the crystal structures 725:746 the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate 725:850 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 7 19 theme double 1331:1336 arg1 mutant 1338:1343 the S530T/G533V double mutant 1315:1343 the S530T/G533V double mutant 1315:1343 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 6 20 theme observed 1004:1011 arg1 binding 1013:1019 the observed binding 1000:1019 the observed binding 1000:1019 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 1 21 theme cyclooxygenase 200:213 arg1 enzymes 215:221 the cyclooxygenase enzymes 196:221 the cyclooxygenase enzymes (COX-1 and COX-2) 196:239 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 1 21 theme cyclooxygenase 200:213 arg1 COX-2 234:238 COX-2 234:238 COX-2 234:238 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 1 21 theme cyclooxygenase 200:213 arg1 COX-1 224:228 COX-1 224:228 COX-1 224:228 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 6 22 dep pose 1021:1024 arg1 2 997:997 2 997:997 2 997:997 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 6 23 theme S530T 1152:1156 arg1 structure 1174:1182 the S530T muCOX-2 crystal structure 1148:1182 the S530T muCOX-2 crystal structure 1148:1182 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 6 24 theme prior 1086:1090 arg1 complex 1078:1084 the enzyme-inhibitor complex 1057:1084 the enzyme-inhibitor complex prior to acetylation 1057:1105 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 4 25 theme oxygen 674:679 arg1 addition 662:669 the addition 658:669 the addition of oxygen to the prostaglandin product 658:708 Ser-530 has also been shown to influence the stereochemistry for the addition of oxygen to the prostaglandin product. 26859324 0 26 theme Crystal 0:6 arg1 Structure 8:16 Crystal Structure 0:16 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 0:132 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 8 27 theme differential 1454:1465 arg1 acetylation 1467:1477 differential acetylation 1454:1477 differential acetylation of COX-2 purified in various detergent systems and nanodiscs 1454:1538 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 6 28 theme complex 1078:1084 arg1 reflective 1043:1052 reflective 1043:1052 reflective 1043:1052 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 0 29 theme Aspirin-Acetylated 21:38 arg1 Cyclooxygenase-2 46:61 Aspirin-Acetylated Human Cyclooxygenase-2 21:61 Aspirin-Acetylated Human Cyclooxygenase-2 21:61 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 0 30 with Products 94:101 arg1 Stereochemistry 117:131 Reversed Stereochemistry 108:131 Reversed Stereochemistry 108:131 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 3 31 theme -hydroxyeicosatetraenoic 551:574 arg1 acid 576:579 the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid 519:579 the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE) 519:590 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 3 31 theme -hydroxyeicosatetraenoic 551:574 arg1 15R-HETE 582:589 15R-HETE 582:589 15R-HETE 582:589 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 5 32 theme COX-2 806:810 arg1 structures 737:746 the crystal structures 725:746 the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate 725:850 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 8 33 theme lipid 1571:1575 arg1 binding 1577:1583 lipid binding 1571:1583 lipid binding 1571:1583 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 3 34 theme acid 576:579 arg1 generation 505:514 the generation 501:514 the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE) 501:590 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 8 35 theme acetylation 1657:1667 arg1 reaction 1669:1676 the acetylation reaction 1653:1676 the acetylation reaction 1653:1676 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 3 36 theme monooxygenated 523:536 arg1 acid 576:579 the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid 519:579 the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE) 519:590 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 3 36 theme monooxygenated 523:536 arg1 15R-HETE 582:589 15R-HETE 582:589 15R-HETE 582:589 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 0 37 theme Cyclooxygenase-2 46:61 arg1 Structure 8:16 Crystal Structure 0:16 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 0:132 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 6 38 theme hydrophobic 976:986 arg1 groove 988:993 the hydrophobic groove 972:993 the hydrophobic groove 972:993 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 1 39 dep enzymes 215:221 arg1 enzymes 215:221 the cyclooxygenase enzymes 196:221 the cyclooxygenase enzymes (COX-1 and COX-2) 196:239 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 1 39 dep enzymes 215:221 arg1 COX-2 234:238 COX-2 234:238 COX-2 234:238 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 1 39 dep enzymes 215:221 arg1 COX-1 224:228 COX-1 224:228 COX-1 224:228 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 7 40 theme aspirin-acetylated 1412:1429 arg1 COX-2 1431:1435 aspirin-acetylated COX-2 1412:1435 aspirin-acetylated COX-2 1412:1435 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 5 41 theme huCOX-2 817:823 arg1 structures 737:746 the crystal structures 725:746 the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate 725:850 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 0 42 theme Human 40:44 arg1 Cyclooxygenase-2 46:61 Aspirin-Acetylated Human Cyclooxygenase-2 21:61 Aspirin-Acetylated Human Cyclooxygenase-2 21:61 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 8 43 theme purified 1488:1495 arg1 COX-2 1482:1486 COX-2 purified 1482:1495 COX-2 purified 1482:1495 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 3 44 theme complete 436:443 arg1 loss 445:448 complete loss 436:448 complete loss of activity 436:460 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 6 45 theme acetylated 925:934 arg1 Ser-530 936:942 the acetylated Ser-530 921:942 the acetylated Ser-530 921:942 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 5 46 theme S530T 751:755 arg1 COX-2 769:773 S530T murine (mu) COX-2 751:773 S530T murine (mu) COX-2 751:773 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 3 47 theme COX-1 421:425 arg1 Acetylation 406:416 Acetylation 406:416 Acetylation of COX-1 406:425 Acetylation of COX-1 leads to complete loss of activity, while acetylation of COX-2 results in the generation of the monooxygenated product 15(R)-hydroxyeicosatetraenoic acid (15R-HETE). 26859324 8 48 theme COX-2 1482:1486 arg1 acetylation 1467:1477 differential acetylation 1454:1477 differential acetylation of COX-2 purified in various detergent systems and nanodiscs 1454:1538 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 6 49 theme hydrophobic 1214:1224 arg1 groove 1226:1231 the hydrophobic groove 1210:1231 the hydrophobic groove 1210:1231 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 5 50 from structures 737:746 arg1 complex 828:834 complex 828:834 complex with salicylate 828:850 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 6 51 theme Thr-530 1129:1135 arg1 rotamer 1137:1143 the observed Thr-530 rotamer 1116:1143 the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure 1116:1182 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 8 52 theme reaction 1669:1676 arg1 rate 1645:1648 the rate 1641:1648 the rate of the acetylation reaction 1641:1676 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 5 53 theme human 795:799 arg1 hu 802:803 hu 802:803 hu 802:803 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 5 53 theme human 795:799 arg1 COX-2 806:810 aspirin-acetylated human (hu) COX-2 776:810 aspirin-acetylated human (hu) COX-2 776:810 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 8 54 theme various 1500:1506 arg1 systems 1518:1524 various detergent systems 1500:1524 various detergent systems 1500:1524 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 1 55 theme anti-inflammatory 165:181 arg1 drugs 183:187 other nonsteroidal anti-inflammatory drugs 146:187 other nonsteroidal anti-inflammatory drugs 146:187 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 6 56 theme observed 1120:1127 arg1 rotamer 1137:1143 the observed Thr-530 rotamer 1116:1143 the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure 1116:1182 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 5 57 with complex 828:834 arg1 salicylate 841:850 salicylate 841:850 salicylate 841:850 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 1 58 theme nonsteroidal 152:163 arg1 drugs 183:187 other nonsteroidal anti-inflammatory drugs 146:187 other nonsteroidal anti-inflammatory drugs 146:187 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 6 59 theme enzyme-inhibitor 1061:1076 arg1 complex 1078:1084 the enzyme-inhibitor complex 1057:1084 the enzyme-inhibitor complex prior to acetylation 1057:1105 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 7 60 dep observations 1267:1278 arg1 basis 1241:1245 basis 1241:1245 basis 1241:1245 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 7 60 dep observations 1267:1278 arg1 the 1237:1239 the 1237:1239 the 1237:1239 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 7 61 theme structural 1256:1265 arg1 observations 1267:1278 these structural observations 1250:1278 these structural observations 1250:1278 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 1 62 theme other 146:150 arg1 drugs 183:187 other nonsteroidal anti-inflammatory drugs 146:187 other nonsteroidal anti-inflammatory drugs 146:187 Aspirin and other nonsteroidal anti-inflammatory drugs target the cyclooxygenase enzymes (COX-1 and COX-2) to block the formation of prostaglandins. 26859324 8 63 theme detergent 1508:1516 arg1 systems 1518:1524 various detergent systems 1500:1524 various detergent systems 1500:1524 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 5 64 theme murine 757:762 arg1 COX-2 769:773 S530T murine (mu) COX-2 751:773 S530T murine (mu) COX-2 751:773 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 7 65 theme 15R-HETE 1400:1407 arg1 generation 1386:1395 the generation 1382:1395 the generation of 15R-HETE by aspirin-acetylated COX-2 1382:1435 On the basis of these structural observations, along with functional analysis of the S530T/G533V double mutant, we propose a working hypothesis for the generation of 15R-HETE by aspirin-acetylated COX-2. 26859324 0 66 dep Structure 8:16 arg1 Insight 64:70 Insight 64:70 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 0:132 Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. 26859324 2 67 theme active 393:398 arg1 site 400:403 the cyclooxygenase active site 374:403 the cyclooxygenase active site 374:403 Aspirin is unique in that it covalently modifies each enzyme by acetylating Ser-530 within the cyclooxygenase active site. 26859324 6 68 from rotamer 1137:1143 arg1 structure 1174:1182 the S530T muCOX-2 crystal structure 1148:1182 the S530T muCOX-2 crystal structure 1148:1182 The structures reveal that (1) the acetylated Ser-530 completely blocks access to the hydrophobic groove, (2) the observed binding pose of salicylate is reflective of the enzyme-inhibitor complex prior to acetylation, and (3) the observed Thr-530 rotamer in the S530T muCOX-2 crystal structure does not impede access to the hydrophobic groove. 26859324 5 69 theme aspirin-acetylated 776:793 arg1 hu 802:803 hu 802:803 hu 802:803 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 5 69 theme aspirin-acetylated 776:793 arg1 COX-2 806:810 aspirin-acetylated human (hu) COX-2 776:810 aspirin-acetylated human (hu) COX-2 776:810 We determined the crystal structures of S530T murine (mu) COX-2, aspirin-acetylated human (hu) COX-2, and huCOX-2 in complex with salicylate to 1.9, 2.0, and 2.4 Å, respectively. 26859324 2 70 theme cyclooxygenase 378:391 arg1 site 400:403 the cyclooxygenase active site 374:403 the cyclooxygenase active site 374:403 Aspirin is unique in that it covalently modifies each enzyme by acetylating Ser-530 within the cyclooxygenase active site. 26859324 8 71 theme membrane-binding 1596:1611 arg1 domain 1613:1618 the membrane-binding domain 1592:1618 the membrane-binding domain of the enzyme 1592:1632 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 26859324 8 71 theme membrane-binding 1596:1611 arg1 enzyme 1627:1632 the enzyme 1623:1632 the enzyme 1623:1632 We also observe differential acetylation of COX-2 purified in various detergent systems and nanodiscs, indicating that detergent and lipid binding within the membrane-binding domain of the enzyme alters the rate of the acetylation reaction in vitro. 27226593 0 0 theme Peroxide 95:102 arg1 Tone 104:107 Peroxide Tone 95:107 Peroxide Tone 95:107 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives Is Dependent on Peroxide Tone. 27226593 7 1 theme substrate-selective 1462:1480 arg1 acid 1453:1456 tolfenamic acid 1442:1456 tolfenamic acid 1442:1456 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 1 theme substrate-selective 1462:1480 arg1 inhibitors 1482:1491 substrate-selective inhibitors 1462:1491 substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety 1462:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 1 theme substrate-selective 1462:1480 arg1 acid 1416:1419 flufenamic acid 1405:1419 flufenamic acid 1405:1419 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 1 theme substrate-selective 1462:1480 arg1 acid 1432:1435 mefenamic acid 1422:1435 mefenamic acid 1422:1435 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 9 2 theme inhibition 1835:1844 arg1 component 1802:1810 an important mechanistic component 1777:1810 an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid 1777:1900 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 9 2 theme inhibition 1835:1844 arg1 tone 1769:1772 peroxide tone 1760:1772 peroxide tone 1760:1772 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 1 3 theme arachidonic 164:174 arg1 AA 182:183 AA 182:183 AA 182:183 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 1 3 theme arachidonic 164:174 arg1 acid 176:179 arachidonic acid 164:179 arachidonic acid (AA) 164:184 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 1 4 theme unique 242:247 arg1 junction 249:256 a unique junction 240:256 a unique junction between the eicosanoid and endocannabinoid signaling pathways 240:318 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 7 5 theme flufenamic 1405:1414 arg1 acid 1453:1456 tolfenamic acid 1442:1456 tolfenamic acid 1442:1456 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 5 theme flufenamic 1405:1414 arg1 inhibitors 1482:1491 substrate-selective inhibitors 1462:1491 substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety 1462:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 5 theme flufenamic 1405:1414 arg1 acid 1416:1419 flufenamic acid 1405:1419 flufenamic acid 1405:1419 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 5 theme flufenamic 1405:1414 arg1 acid 1432:1435 mefenamic acid 1422:1435 mefenamic acid 1422:1435 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 2 6 dep active 444:449 arg1 such 401:404 such 401:404 such 401:404 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 1 7 theme acid 176:179 arg1 oxygenation 149:159 the oxygenation 145:159 the oxygenation of arachidonic acid (AA) and endocannabinoid substrates 145:215 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 8 8 theme lipid 1670:1674 arg1 acid 1716:1719 the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1666:1719 the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1666:1719 Substrate-selective inhibition was attenuated by the addition of the lipid peroxide 15-hydroperoxyeicosatertaenoic acid. 27226593 9 9 theme mefenamic 1866:1874 arg1 acid 1876:1879 mefenamic acid 1866:1879 mefenamic acid 1866:1879 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 3 10 theme sufficient 668:677 arg1 monomer 660:666 a single monomer 651:666 a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid 651:749 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 6 11 theme carboxylate 1202:1212 arg1 group 1214:1218 the carboxylate group 1198:1218 the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel 1198:1281 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 6 12 theme drug 1064:1067 arg1 structures 1045:1054 Crystal structures 1037:1054 Crystal structures of each drug in complex with human COX-2 1037:1095 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 7 13 theme heme 1588:1591 arg1 moiety 1593:1598 the heme moiety 1584:1598 the heme moiety 1584:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 14 theme tolfenamic 1442:1451 arg1 acid 1453:1456 tolfenamic acid 1442:1456 tolfenamic acid 1442:1456 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 14 theme tolfenamic 1442:1451 arg1 inhibitors 1482:1491 substrate-selective inhibitors 1462:1491 substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety 1462:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 14 theme tolfenamic 1442:1451 arg1 acid 1416:1419 flufenamic acid 1405:1419 flufenamic acid 1405:1419 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 14 theme tolfenamic 1442:1451 arg1 acid 1432:1435 mefenamic acid 1422:1435 mefenamic acid 1422:1435 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 6 15 from structures 1045:1054 arg1 complex 1072:1078 complex 1072:1078 complex with human COX-2 1072:1095 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 3 16 theme reversible 482:491 arg1 NSAIDs 544:549 NSAIDs 544:549 NSAIDs 544:549 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 3 16 theme reversible 482:491 arg1 drugs 537:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs 468:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) 468:550 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 7 17 theme moiety 1593:1598 arg1 states 1574:1579 higher oxidation states 1557:1579 higher oxidation states of the heme moiety 1557:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 3 18 theme endocannabinoids 709:724 arg1 oxygenation 694:704 the oxygenation 690:704 the oxygenation of endocannabinoids 690:724 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 6 19 theme channel 1275:1281 arg1 top 1264:1266 the top 1260:1266 the top of the channel 1260:1281 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 5 20 used utilized 849:856 arg2 We 846:847 We 846:847 We 846:847 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 2 21 dep displays 367:374 arg1 active 444:449 active 444:449 active 444:449 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 6 22 theme human 1085:1089 arg1 COX-2 1091:1095 human COX-2 1085:1095 human COX-2 1085:1095 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 5 23 theme biophysical 873:883 arg1 methods 885:891 structural and biophysical methods 858:891 structural and biophysical methods 858:891 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 8 24 theme 15-hydroperoxyeicosatertaenoic 1685:1714 arg1 acid 1716:1719 the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1666:1719 the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1666:1719 Substrate-selective inhibition was attenuated by the addition of the lipid peroxide 15-hydroperoxyeicosatertaenoic acid. 27226593 2 25 theme given 456:460 arg1 time 462:465 a given time 454:465 a given time 454:465 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 7 26 theme spin 1344:1347 arg1 resonance 1349:1357 continuous-wave electron spin resonance 1319:1357 continuous-wave electron spin resonance 1319:1357 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 0 27 theme Substrate-selective 0:18 arg1 Inhibition 20:29 Substrate-selective Inhibition 0:29 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives 0:77 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives Is Dependent on Peroxide Tone. 27226593 7 28 theme UV-visible 1364:1373 arg1 spectroscopy 1375:1386 UV-visible spectroscopy 1364:1386 UV-visible spectroscopy 1364:1386 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 9 29 theme tolfenamic 1886:1895 arg1 acid 1897:1900 tolfenamic acid 1886:1900 tolfenamic acid 1886:1900 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 9 30 theme peroxide 1760:1767 arg1 component 1802:1810 an important mechanistic component 1777:1810 an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid 1777:1900 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 9 30 theme peroxide 1760:1767 arg1 tone 1769:1772 peroxide tone 1760:1772 peroxide tone 1760:1772 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 3 31 dep reversible 482:491 arg1 competitive 494:504 competitive 494:504 competitive 494:504 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 6 32 theme cyclooxygenase 1142:1155 arg1 channel 1157:1163 the cyclooxygenase channel 1138:1163 the cyclooxygenase channel in an inverted orientation 1138:1190 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 2 33 theme dimer 435:439 arg1 dimer 435:439 the dimer 431:439 the dimer 431:439 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 2 33 theme dimer 435:439 arg1 monomer 420:426 only one monomer 411:426 only one monomer of the dimer 411:439 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 3 34 theme arachidonic 734:744 arg1 acid 746:749 arachidonic acid 734:749 a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid 651:749 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 9 35 theme flufenamic 1849:1858 arg1 acid 1860:1863 flufenamic acid 1849:1863 flufenamic acid 1849:1863 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 6 36 theme inverted 1171:1178 arg1 orientation 1180:1190 an inverted orientation 1168:1190 an inverted orientation 1168:1190 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 2 37 theme half-of-site 376:387 arg1 reactivity 389:398 half-of-site reactivity 376:398 half-of-site reactivity 376:398 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 0 38 theme Cyclooxygeanse-2 34:49 arg1 Inhibition 20:29 Substrate-selective Inhibition 0:29 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives 0:77 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives Is Dependent on Peroxide Tone. 27226593 5 39 theme tolfenamic 961:970 arg1 acid 972:975 tolfenamic acid 961:975 tolfenamic acid 961:975 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 1 40 theme endocannabinoid 190:204 arg1 substrates 206:215 endocannabinoid substrates 190:215 endocannabinoid substrates 190:215 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 1 41 theme endocannabinoid 285:299 arg1 pathways 311:318 the eicosanoid and endocannabinoid signaling pathways 266:318 pathways 311:318 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 3 42 theme anti-inflammatory 519:535 arg1 NSAIDs 544:549 NSAIDs 544:549 NSAIDs 544:549 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 3 42 theme anti-inflammatory 519:535 arg1 drugs 537:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs 468:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) 468:550 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 5 43 theme meclofenamic 922:933 arg1 acid 935:938 meclofenamic acid 922:938 meclofenamic acid 922:938 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 8 44 theme Substrate-selective 1601:1619 arg1 inhibition 1621:1630 Substrate-selective inhibition 1601:1630 Substrate-selective inhibition 1601:1630 Substrate-selective inhibition was attenuated by the addition of the lipid peroxide 15-hydroperoxyeicosatertaenoic acid. 27226593 1 45 theme substrates 206:215 arg1 oxygenation 149:159 the oxygenation 145:159 the oxygenation of arachidonic acid (AA) and endocannabinoid substrates 145:215 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 0 46 theme Fenamic 54:60 arg1 Derivatives 67:77 Fenamic Acid Derivatives 54:77 Fenamic Acid Derivatives 54:77 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives Is Dependent on Peroxide Tone. 27226593 1 47 theme signaling 301:309 arg1 pathways 311:318 the eicosanoid and endocannabinoid signaling pathways 266:318 pathways 311:318 Cyclooxygenase-2 (COX-2) catalyzes the oxygenation of arachidonic acid (AA) and endocannabinoid substrates, placing the enzyme at a unique junction between the eicosanoid and endocannabinoid signaling pathways. 27226593 9 48 theme important 1780:1788 arg1 component 1802:1810 an important mechanistic component 1777:1810 an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid 1777:1900 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 9 48 theme important 1780:1788 arg1 tone 1769:1772 peroxide tone 1760:1772 peroxide tone 1760:1772 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 2 49 from time 462:465 arg1 active 444:449 active 444:449 active 444:449 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 7 50 theme mefenamic 1422:1430 arg1 acid 1453:1456 tolfenamic acid 1442:1456 tolfenamic acid 1442:1456 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 50 theme mefenamic 1422:1430 arg1 inhibitors 1482:1491 substrate-selective inhibitors 1462:1491 substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety 1462:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 50 theme mefenamic 1422:1430 arg1 acid 1416:1419 flufenamic acid 1405:1419 flufenamic acid 1405:1419 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 50 theme mefenamic 1422:1430 arg1 acid 1432:1435 mefenamic acid 1422:1435 mefenamic acid 1422:1435 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 5 51 theme structural 858:867 arg1 methods 885:891 structural and biophysical methods 858:891 structural and biophysical methods 858:891 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 9 52 theme mechanistic 1790:1800 arg1 component 1802:1810 an important mechanistic component 1777:1810 an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid 1777:1900 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 9 52 theme mechanistic 1790:1800 arg1 tone 1769:1772 peroxide tone 1760:1772 peroxide tone 1760:1772 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 7 53 theme oxidation 1564:1572 arg1 states 1574:1579 higher oxidation states 1557:1579 higher oxidation states of the heme moiety 1557:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 7 54 theme continuous-wave 1319:1333 arg1 resonance 1349:1357 continuous-wave electron spin resonance 1319:1357 continuous-wave electron spin resonance 1319:1357 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 6 55 with complex 1072:1078 arg1 COX-2 1091:1095 human COX-2 1085:1095 human COX-2 1085:1095 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 7 56 theme fluorescence 1295:1306 arg1 quenching 1308:1316 Tryptophan fluorescence quenching 1284:1316 Tryptophan fluorescence quenching 1284:1316 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 3 57 theme Certain 468:474 arg1 NSAIDs 544:549 NSAIDs 544:549 NSAIDs 544:549 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 3 57 theme Certain 468:474 arg1 drugs 537:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs 468:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) 468:550 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 0 58 theme Acid 62:65 arg1 Derivatives 67:77 Fenamic Acid Derivatives 54:77 Fenamic Acid Derivatives 54:77 Substrate-selective Inhibition of Cyclooxygeanse-2 by Fenamic Acid Derivatives Is Dependent on Peroxide Tone. 27226593 7 59 theme higher 1557:1562 arg1 states 1574:1579 higher oxidation states 1557:1579 higher oxidation states of the heme moiety 1557:1598 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 6 60 from channel 1157:1163 arg1 orientation 1180:1190 an inverted orientation 1168:1190 an inverted orientation 1168:1190 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 3 61 theme rapid 476:480 arg1 NSAIDs 544:549 NSAIDs 544:549 NSAIDs 544:549 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 3 61 theme rapid 476:480 arg1 drugs 537:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs 468:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) 468:550 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 4 62 theme substrate-selective 793:811 arg1 inhibition 813:822 substrate-selective inhibition 793:822 substrate-selective inhibition 793:822 The underlying mechanism responsible for substrate-selective inhibition has remained elusive. 27226593 9 63 theme substrate-selective 1815:1833 arg1 inhibition 1835:1844 substrate-selective inhibition 1815:1844 substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid 1815:1900 Collectively, these studies implicate peroxide tone as an important mechanistic component of substrate-selective inhibition by flufenamic acid, mefenamic acid, and tolfenamic acid. 27226593 3 64 theme single 653:658 arg1 monomer 660:666 a single monomer 651:666 a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid 651:749 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 2 65 theme sequence 332:339 arg1 homodimer 341:349 a sequence homodimer 330:349 a sequence homodimer 330:349 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 2 65 theme sequence 332:339 arg1 COX-2 321:325 COX-2 321:325 COX-2 321:325 COX-2 is a sequence homodimer, but the enzyme displays half-of-site reactivity, such that only one monomer of the dimer is active at a given time. 27226593 4 66 theme underlying 756:765 arg1 mechanism 767:775 The underlying mechanism 752:775 The underlying mechanism responsible for substrate-selective inhibition 752:822 The underlying mechanism responsible for substrate-selective inhibition has remained elusive. 27226593 3 67 theme inhibitor 638:646 arg1 binding 627:633 the binding 623:633 the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid 623:749 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 4 68 theme responsible 777:787 arg1 mechanism 767:775 The underlying mechanism 752:775 The underlying mechanism responsible for substrate-selective inhibition 752:822 The underlying mechanism responsible for substrate-selective inhibition has remained elusive. 27226593 7 69 theme Tryptophan 1284:1293 arg1 quenching 1308:1316 Tryptophan fluorescence quenching 1284:1316 Tryptophan fluorescence quenching 1284:1316 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 5 70 theme mefenamic 941:949 arg1 acid 951:954 mefenamic acid 941:954 mefenamic acid 941:954 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 5 71 theme substrate-selective 1005:1023 arg1 inhibitors 1025:1034 substrate-selective inhibitors 1005:1034 substrate-selective inhibitors 1005:1034 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 7 72 theme electron 1335:1342 arg1 resonance 1349:1357 continuous-wave electron spin resonance 1319:1357 continuous-wave electron spin resonance 1319:1357 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 5 73 theme flufenamic 905:914 arg1 acid 916:919 flufenamic acid 905:919 flufenamic acid 905:919 We utilized structural and biophysical methods to evaluate flufenamic acid, meclofenamic acid, mefenamic acid, and tolfenamic acid for their ability to act as substrate-selective inhibitors. 27226593 6 74 theme Crystal 1037:1043 arg1 structures 1045:1054 Crystal structures 1037:1054 Crystal structures of each drug in complex with human COX-2 1037:1095 Crystal structures of each drug in complex with human COX-2 revealed that the inhibitor binds within the cyclooxygenase channel in an inverted orientation, with the carboxylate group interacting with Tyr-385 and Ser-530 at the top of the channel. 27226593 8 75 theme acid 1716:1719 arg1 addition 1654:1661 the addition 1650:1661 the addition of the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1650:1719 Substrate-selective inhibition was attenuated by the addition of the lipid peroxide 15-hydroperoxyeicosatertaenoic acid. 27226593 7 76 theme tyrosyl 1528:1534 arg1 radicals 1536:1543 tyrosyl radicals 1528:1543 tyrosyl radicals 1528:1543 Tryptophan fluorescence quenching, continuous-wave electron spin resonance, and UV-visible spectroscopy demonstrate that flufenamic acid, mefenamic acid, and tolfenamic acid are substrate-selective inhibitors that bind rapidly to COX-2, quench tyrosyl radicals, and reduce higher oxidation states of the heme moiety. 27226593 3 77 theme substrate-selective 590:608 arg1 manner 610:615 a substrate-selective manner 588:615 a substrate-selective manner 588:615 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 3 78 theme nonsteroidal 506:517 arg1 NSAIDs 544:549 NSAIDs 544:549 NSAIDs 544:549 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 3 78 theme nonsteroidal 506:517 arg1 drugs 537:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs 468:541 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) 468:550 Certain rapid reversible, competitive nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit COX-2 in a substrate-selective manner, with the binding of inhibitor to a single monomer sufficient to inhibit the oxygenation of endocannabinoids but not arachidonic acid. 27226593 8 79 theme peroxide 1676:1683 arg1 acid 1716:1719 the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1666:1719 the lipid peroxide 15-hydroperoxyeicosatertaenoic acid 1666:1719 Substrate-selective inhibition was attenuated by the addition of the lipid peroxide 15-hydroperoxyeicosatertaenoic acid. 27710942 7 0 theme channel 1068:1074 arg1 pocket 1039:1044 the side pocket 1030:1044 the side pocket of the cyclooxygenase channel 1030:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 4 1 theme sulfonamide 590:600 arg1 moiety 602:607 the sulfonamide moiety 586:607 the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib 586:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 5 2 theme new 671:673 arg1 conditions 691:700 new crystallization conditions 671:700 new crystallization conditions 671:700 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 1 3 theme inhibitors 138:147 arg1 one 86:88 one 86:88 one 86:88 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 1 3 theme inhibitors 138:147 arg1 coxibs 150:155 coxibs 150:155 coxibs 150:155 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 1 3 theme inhibitors 138:147 arg1 inhibitors 138:147 the first selective cyclooxygenase-2 (COX-2) inhibitors 93:147 the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) 93:156 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 6 4 theme level 912:916 arg1 details 918:924 the first atomic level details 895:924 the first atomic level details of the binding of Vioxx to COX-2 895:957 The crystal structure provides the first atomic level details of the binding of Vioxx to COX-2. 27710942 7 5 theme isoform 1103:1109 arg1 selectivity 1111:1121 the isoform selectivity 1099:1121 the isoform selectivity of this drug 1099:1134 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 7 6 theme located 1019:1025 arg1 moiety 1012:1017 its methyl sulfone moiety 993:1017 its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel 993:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 5 7 theme 2.7 Å 846:850 arg1 resolution 852:861 2.7 Å resolution 846:861 2.7 Å resolution 846:861 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 7 8 theme methyl 997:1002 arg1 moiety 1012:1017 its methyl sulfone moiety 993:1017 its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel 993:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 2 9 theme drug 330:333 arg1 effects 315:321 the adverse cardiovascular effects 288:321 the adverse cardiovascular effects of the drug 288:333 Within five years after its release to the public, Vioxx was withdrawn from the market owing to the adverse cardiovascular effects of the drug. 27710942 6 10 theme atomic 905:910 arg1 details 918:924 the first atomic level details 895:924 the first atomic level details of the binding of Vioxx to COX-2 895:957 The crystal structure provides the first atomic level details of the binding of Vioxx to COX-2. 27710942 7 11 theme sulfone 1004:1010 arg1 moiety 1012:1017 its methyl sulfone moiety 993:1017 its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel 993:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 3 12 theme development 376:386 arg1 knowledge 359:367 the widespread knowledge 344:367 the widespread knowledge of the development and withdrawal of Vioxx 344:410 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 4 13 located found 609:613 arg1 valdecoxib 653:662 valdecoxib 653:662 valdecoxib 653:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 4 13 located found 609:613 arg1 coxibs 624:629 other coxibs 618:629 other coxibs such as celecoxib and valdecoxib 618:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 4 13 located found 609:613 arg1 celecoxib 639:647 celecoxib 639:647 celecoxib 639:647 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 4 13 located found 609:613 arg2 moiety 602:607 the sulfonamide moiety 586:607 the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib 586:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 0 14 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of rofecoxib 0:29 Crystal structure of rofecoxib bound to human cyclooxygenase-2. 27710942 6 15 theme first 899:903 arg1 details 918:924 the first atomic level details 895:924 the first atomic level details of the binding of Vioxx to COX-2 895:957 The crystal structure provides the first atomic level details of the binding of Vioxx to COX-2. 27710942 0 16 theme rofecoxib 21:29 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of rofecoxib 0:29 Crystal structure of rofecoxib bound to human cyclooxygenase-2. 27710942 3 17 theme molecular 447:455 arg1 level 457:461 the molecular level 443:461 the molecular level 443:461 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 3 18 theme Vioxx 406:410 arg1 withdrawal 392:401 withdrawal 392:401 withdrawal 392:401 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 3 18 theme Vioxx 406:410 arg1 development 376:386 development 376:386 development 376:386 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 5 19 theme crystallization 675:689 arg1 conditions 691:700 new crystallization conditions 671:700 new crystallization conditions 671:700 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 5 20 theme human 763:767 arg1 COX-2 769:773 human COX-2 763:773 human COX-2 763:773 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 4 21 theme sulfone 559:565 arg1 moiety 567:572 a methyl sulfone moiety 550:572 a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib 550:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 3 22 theme widespread 348:357 arg1 knowledge 359:367 the widespread knowledge 344:367 the widespread knowledge of the development and withdrawal of Vioxx 344:410 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 6 23 theme crystal 868:874 arg1 structure 876:884 The crystal structure 864:884 The crystal structure 864:884 The crystal structure provides the first atomic level details of the binding of Vioxx to COX-2. 27710942 4 24 contain contains 541:548 arg2 moiety 567:572 a methyl sulfone moiety 550:572 a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib 550:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 4 24 contain contains 541:548 arg1 inhibitor 531:539 the inhibitor 527:539 the inhibitor 527:539 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 7 25 theme cyclooxygenase 1053:1066 arg1 channel 1068:1074 the cyclooxygenase channel 1049:1074 the cyclooxygenase channel 1049:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 5 26 theme COX-2 769:773 arg1 determination 746:758 the structural determination 731:758 the structural determination of human COX-2 731:773 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 4 27 theme methyl 552:557 arg1 moiety 567:572 a methyl sulfone moiety 550:572 a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib 550:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 7 28 theme side 1034:1037 arg1 pocket 1039:1044 the side pocket 1030:1044 the side pocket of the cyclooxygenase channel 1030:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 7 29 from located 1019:1025 arg1 pocket 1039:1044 the side pocket 1030:1044 the side pocket of the cyclooxygenase channel 1030:1074 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 1 30 from use 177:179 arg1 humans 184:189 humans 184:189 humans 184:189 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 4 31 theme other 618:622 arg1 valdecoxib 653:662 valdecoxib 653:662 valdecoxib 653:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 4 31 theme other 618:622 arg1 coxibs 624:629 other coxibs 618:629 other coxibs such as celecoxib and valdecoxib 618:662 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 4 31 theme other 618:622 arg1 celecoxib 639:647 celecoxib 639:647 celecoxib 639:647 Vioxx is unique in that the inhibitor contains a methyl sulfone moiety in place of the sulfonamide moiety found in other coxibs such as celecoxib and valdecoxib. 27710942 0 32 theme human 40:44 arg1 cyclooxygenase-2 46:61 human cyclooxygenase-2 40:61 human cyclooxygenase-2 40:61 Crystal structure of rofecoxib bound to human cyclooxygenase-2. 27710942 3 33 dep development 376:386 arg1 the 372:374 the 372:374 the 372:374 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 1 34 theme first 97:101 arg1 COX-2 131:135 COX-2 131:135 COX-2 131:135 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 1 34 theme first 97:101 arg1 cyclooxygenase-2 113:128 first selective cyclooxygenase-2 97:128 the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) 93:156 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 7 35 from pocket 1039:1044 arg1 located 1019:1025 located 1019:1025 located 1019:1025 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 1 36 theme selective 103:111 arg1 COX-2 131:135 COX-2 131:135 COX-2 131:135 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 1 36 theme selective 103:111 arg1 cyclooxygenase-2 113:128 first selective cyclooxygenase-2 97:128 the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) 93:156 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 6 37 theme Vioxx 944:948 arg1 binding 933:939 the binding 929:939 the binding of Vioxx to COX-2 929:957 The crystal structure provides the first atomic level details of the binding of Vioxx to COX-2. 27710942 5 38 with complex 778:784 arg1 Vioxx 791:795 Vioxx 791:795 Vioxx 791:795 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 1 39 theme cyclooxygenase-2 113:128 arg1 coxibs 150:155 coxibs 150:155 coxibs 150:155 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 1 39 theme cyclooxygenase-2 113:128 arg1 inhibitors 138:147 the first selective cyclooxygenase-2 (COX-2) inhibitors 93:147 the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) 93:156 Rofecoxib (Vioxx) was one of the first selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) to be approved for use in humans. 27710942 3 40 theme withdrawal 392:401 arg1 knowledge 359:367 the widespread knowledge 344:367 the widespread knowledge of the development and withdrawal of Vioxx 344:410 Despite the widespread knowledge of the development and withdrawal of Vioxx, relatively little is known at the molecular level about how the inhibitor binds to COX-2. 27710942 2 41 theme cardiovascular 300:313 arg1 effects 315:321 the adverse cardiovascular effects 288:321 the adverse cardiovascular effects of the drug 288:333 Within five years after its release to the public, Vioxx was withdrawn from the market owing to the adverse cardiovascular effects of the drug. 27710942 6 42 theme binding 933:939 arg1 details 918:924 the first atomic level details 895:924 the first atomic level details of the binding of Vioxx to COX-2 895:957 The crystal structure provides the first atomic level details of the binding of Vioxx to COX-2. 27710942 7 43 theme drug 1131:1134 arg1 selectivity 1111:1121 the isoform selectivity 1099:1121 the isoform selectivity of this drug 1099:1134 As anticipated, Vioxx binds with its methyl sulfone moiety located in the side pocket of the cyclooxygenase channel, providing support for the isoform selectivity of this drug. 27710942 5 44 theme structural 735:744 arg1 determination 746:758 the structural determination 731:758 the structural determination of human COX-2 731:773 Here, new crystallization conditions were identified that allowed the structural determination of human COX-2 in complex with Vioxx and the structure was subsequently determined to 2.7 Å resolution. 27710942 2 45 theme adverse 292:298 arg1 effects 315:321 the adverse cardiovascular effects 288:321 the adverse cardiovascular effects of the drug 288:333 Within five years after its release to the public, Vioxx was withdrawn from the market owing to the adverse cardiovascular effects of the drug. 23756651 1 0 dep ligases 250:256 arg1 finger 263:268 RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 258:308 finger 263:268 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 0 dep ligases 250:256 arg1 ligases 250:256 the transmembrane E3 ubiquitin ligases 219:256 the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 219:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 0 dep ligases 250:256 arg1 finger 287:292 RING finger 3 282:294 RING finger 3 (RNF43/ZNRF3) 282:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 0 dep ligases 250:256 arg1 RNF43/ZNRF3 297:307 RNF43/ZNRF3 297:307 RNF43/ZNRF3 297:307 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 4 1 theme RSPO1 610:614 arg1 affinity 598:605 the affinity 594:605 the affinity of RSPO1 to RNF43 594:623 LGR5 does not contact RNF43 but increases the affinity of RSPO1 to RNF43, supporting LGR5 as an engagement receptor and RNF43 as an effector receptor. 23756651 1 2 theme transmembrane 223:235 arg1 finger 263:268 RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 258:308 finger 263:268 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 2 theme transmembrane 223:235 arg1 ligases 250:256 the transmembrane E3 ubiquitin ligases 219:256 the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 219:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 2 theme transmembrane 223:235 arg1 finger 287:292 RING finger 3 282:294 RING finger 3 (RNF43/ZNRF3) 282:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 4 3 theme effector 684:691 arg1 receptor 693:700 an effector receptor 681:700 an effector receptor 681:700 LGR5 does not contact RNF43 but increases the affinity of RSPO1 to RNF43, supporting LGR5 as an engagement receptor and RNF43 as an effector receptor. 23756651 4 3 theme effector 684:691 arg1 LGR5 637:640 LGR5 637:640 LGR5 637:640 LGR5 does not contact RNF43 but increases the affinity of RSPO1 to RNF43, supporting LGR5 as an engagement receptor and RNF43 as an effector receptor. 23756651 3 4 theme rod 456:458 arg1 module 460:465 its rod module 452:465 its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43 452:549 RSPO1 is sandwiched by LGR5 and RNF43, with its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43. 23756651 3 5 theme cysteine-rich 474:486 arg1 CRD 496:498 CRD 496:498 CRD 496:498 RSPO1 is sandwiched by LGR5 and RNF43, with its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43. 23756651 3 5 theme cysteine-rich 474:486 arg1 domain 488:493 the cysteine-rich domain 470:493 the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43 470:549 RSPO1 is sandwiched by LGR5 and RNF43, with its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43. 23756651 1 6 theme E3 237:238 arg1 finger 263:268 RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 258:308 finger 263:268 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 6 theme E3 237:238 arg1 ligases 250:256 the transmembrane E3 ubiquitin ligases 219:256 the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 219:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 6 theme E3 237:238 arg1 finger 287:292 RING finger 3 282:294 RING finger 3 (RNF43/ZNRF3) 282:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 3 7 theme domain 488:493 arg1 module 460:465 its rod module 452:465 its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43 452:549 RSPO1 is sandwiched by LGR5 and RNF43, with its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43. 23756651 1 8 theme ubiquitin 240:248 arg1 finger 263:268 RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 258:308 finger 263:268 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 8 theme ubiquitin 240:248 arg1 ligases 250:256 the transmembrane E3 ubiquitin ligases 219:256 the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 219:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 8 theme ubiquitin 240:248 arg1 finger 287:292 RING finger 3 282:294 RING finger 3 (RNF43/ZNRF3) 282:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 5 9 theme therapeutic 770:780 arg1 targeting 782:790 therapeutic targeting 770:790 therapeutic targeting 770:790 Disease mutations map to the RSPO1-RNF43 interface, which promises therapeutic targeting. 23756651 0 10 theme structural 4:13 arg1 basis 15:19 The structural basis 0:19 The structural basis of R-spondin recognition 0:44 The structural basis of R-spondin recognition by LGR5 and RNF43. 23756651 1 11 theme leucine-rich 142:153 arg1 receptors 191:199 leucine-rich repeat-containing G-protein-coupled receptors 4-6 142:203 leucine-rich repeat-containing G-protein-coupled receptors 4-6 142:203 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 12 theme repeat-containing 155:171 arg1 receptors 191:199 leucine-rich repeat-containing G-protein-coupled receptors 4-6 142:203 leucine-rich repeat-containing G-protein-coupled receptors 4-6 142:203 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 2 13 theme physical 390:397 arg1 linkage 399:405 their physical linkage 384:405 their physical linkage 384:405 The structure of RSPO1 bound to both LGR5 and RNF43 ectodomains confirms their physical linkage. 23756651 1 14 theme RING 258:261 arg1 finger 263:268 RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 258:308 finger 263:268 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 14 theme RING 258:261 arg1 ligases 250:256 the transmembrane E3 ubiquitin ligases 219:256 the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 219:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 15 theme G-protein-coupled 173:189 arg1 receptors 191:199 leucine-rich repeat-containing G-protein-coupled receptors 4-6 142:203 leucine-rich repeat-containing G-protein-coupled receptors 4-6 142:203 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 4 16 theme engagement 648:657 arg1 receptor 659:666 an engagement receptor 645:666 an engagement receptor 645:666 LGR5 does not contact RNF43 but increases the affinity of RSPO1 to RNF43, supporting LGR5 as an engagement receptor and RNF43 as an effector receptor. 23756651 4 16 theme engagement 648:657 arg1 LGR5 637:640 LGR5 637:640 LGR5 637:640 LGR5 does not contact RNF43 but increases the affinity of RSPO1 to RNF43, supporting LGR5 as an engagement receptor and RNF43 as an effector receptor. 23756651 1 17 theme stem 114:117 arg1 behavior 124:131 stem cell behavior 114:131 stem cell behavior 114:131 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 0 18 theme recognition 34:44 arg1 basis 15:19 The structural basis 0:19 The structural basis of R-spondin recognition 0:44 The structural basis of R-spondin recognition by LGR5 and RNF43. 23756651 3 19 with sandwiched 417:426 arg1 module 460:465 its rod module 452:465 its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43 452:549 RSPO1 is sandwiched by LGR5 and RNF43, with its rod module of the cysteine-rich domain (CRD) contacting LGR5 and a hairpin inserted into RNF43. 23756651 4 20 theme contact 566:572 arg1 RNF43 574:578 contact RNF43 566:578 contact RNF43 566:578 LGR5 does not contact RNF43 but increases the affinity of RSPO1 to RNF43, supporting LGR5 as an engagement receptor and RNF43 as an effector receptor. 23756651 2 21 theme RNF43 357:361 arg1 ectodomains 363:373 RNF43 ectodomains 357:373 RNF43 ectodomains 357:373 The structure of RSPO1 bound to both LGR5 and RNF43 ectodomains confirms their physical linkage. 23756651 0 22 theme R-spondin 24:32 arg1 recognition 34:44 R-spondin recognition 24:44 R-spondin recognition 24:44 The structural basis of R-spondin recognition by LGR5 and RNF43. 23756651 1 23 theme RING 282:285 arg1 ligases 250:256 the transmembrane E3 ubiquitin ligases 219:256 the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 219:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 23 theme RING 282:285 arg1 finger 287:292 RING finger 3 282:294 RING finger 3 (RNF43/ZNRF3) 282:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 23 theme RING 282:285 arg1 RNF43/ZNRF3 297:307 RNF43/ZNRF3 297:307 RNF43/ZNRF3 297:307 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 24 theme Wnt 92:94 arg1 signaling 96:104 Wnt signaling 92:104 Wnt signaling 92:104 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 5 25 theme Disease 703:709 arg1 mutations 711:719 Disease mutations 703:719 Disease mutations 703:719 Disease mutations map to the RSPO1-RNF43 interface, which promises therapeutic targeting. 23756651 0 26 dep basis 15:19 arg1 RNF43 58:62 RNF43 58:62 RNF43 58:62 The structural basis of R-spondin recognition by LGR5 and RNF43. 23756651 0 26 dep basis 15:19 arg1 LGR5 49:52 LGR5 49:52 LGR5 49:52 The structural basis of R-spondin recognition by LGR5 and RNF43. 23756651 1 27 theme cell 119:122 arg1 behavior 124:131 stem cell behavior 114:131 stem cell behavior 114:131 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 5 28 theme RSPO1-RNF43 732:742 arg1 interface 744:752 the RSPO1-RNF43 interface 728:752 the RSPO1-RNF43 interface 728:752 Disease mutations map to the RSPO1-RNF43 interface, which promises therapeutic targeting. 23756651 2 29 theme RSPO1 328:332 arg1 structure 315:323 The structure 311:323 The structure of RSPO1 bound to both LGR5 and RNF43 ectodomains 311:373 The structure of RSPO1 bound to both LGR5 and RNF43 ectodomains confirms their physical linkage. 23756651 1 30 dep finger 263:268 arg1 43/zinc 270:276 43/zinc 270:276 43/zinc 270:276 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 31 dep receptors 191:199 arg1 4-6 201:203 4-6 201:203 4-6 201:203 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 32 dep enhance 84:90 arg1 bind 134:137 bind 134:137 bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3) 134:308 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 23756651 1 32 dep enhance 84:90 arg1 affect 107:112 affect 107:112 affect stem cell behavior 107:131 R-spondins (RSPOs) enhance Wnt signaling, affect stem cell behavior, bind to leucine-rich repeat-containing G-protein-coupled receptors 4-6, (LGR4-6) and the transmembrane E3 ubiquitin ligases RING finger 43/zinc and RING finger 3 (RNF43/ZNRF3). 17005555 0 0 theme central 64:70 arg1 system 80:85 central nervous system 64:85 central nervous system repair inhibition 64:103 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 5 1 theme such 1088:1091 arg1 assembly 1093:1100 such assembly 1088:1100 such assembly 1088:1100 The structure, together with biophysical analysis of its solution properties, reveals that in the crystals and in solution Lingo-1 persistently associates with itself to form a stable tetramer and that it is its LRR-Ig-composite fold that drives such assembly. 17005555 8 2 theme functional 1465:1474 arg1 sites 1484:1488 Potential functional binding sites 1455:1488 Potential functional binding sites 1455:1488 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 2 theme functional 1465:1474 arg1 self-recognition 1562:1577 self-recognition 1562:1577 self-recognition 1562:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 2 theme functional 1465:1474 arg1 site 1554:1557 the site 1550:1557 the site of self-recognition 1550:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 1 3 theme -mediated 125:133 arg1 control 135:141 Nogo receptor (NgR)-mediated control 106:141 Nogo receptor (NgR)-mediated control of axon growth 106:156 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 7 4 theme surface 1296:1302 arg1 area 1304:1307 a large surface area 1288:1307 a large surface area (9,200 A2) 1288:1318 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 7 4 theme surface 1296:1302 arg1 A2 1316:1317 9,200 A2 1310:1317 9,200 A2 1310:1317 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 1 5 theme protein 225:231 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 7 6 theme NgR 1399:1401 arg1 components 1411:1420 the NgR complex components 1395:1420 the NgR complex components 1395:1420 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 6 7 theme LRR 1210:1212 arg1 domain 1214:1219 each LRR domain 1205:1219 each LRR domain filling an open cleft in an adjacent protomer 1205:1265 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 4 8 theme ectodomain 692:701 arg1 structure 660:668 the crystal structure 648:668 the crystal structure of the ligand-binding ectodomain of human Lingo-1 648:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 0 9 theme system 80:85 arg1 inhibition 94:103 central nervous system repair inhibition 64:103 central nervous system repair inhibition 64:103 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 7 10 theme complex 1403:1409 arg1 components 1411:1420 the NgR complex components 1395:1420 the NgR complex components 1395:1420 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 6 11 theme open 1232:1235 arg1 cleft 1237:1241 an open cleft 1229:1241 an open cleft in an adjacent protomer 1229:1265 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 3 12 theme Lingo-1 541:547 arg1 function 549:556 Lingo-1 function 541:556 Lingo-1 function 541:556 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 0 13 theme nervous 72:78 arg1 system 80:85 central nervous system 64:85 central nervous system repair inhibition 64:103 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 8 14 theme Potential 1455:1463 arg1 sites 1484:1488 Potential functional binding sites 1455:1488 Potential functional binding sites 1455:1488 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 14 theme Potential 1455:1463 arg1 self-recognition 1562:1577 self-recognition 1562:1577 self-recognition 1562:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 14 theme Potential 1455:1463 arg1 site 1554:1557 the site 1550:1557 the site of self-recognition 1550:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 15 theme self-recognition 1562:1577 arg1 self-recognition 1562:1577 self-recognition 1562:1577 self-recognition 1562:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 15 theme self-recognition 1562:1577 arg1 site 1554:1557 the site 1550:1557 the site of self-recognition 1550:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 3 16 theme function 549:556 arg1 modulation 527:536 pharmacological modulation 511:536 pharmacological modulation of Lingo-1 function 511:556 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 16 theme function 549:556 arg1 approach 575:582 a novel approach 567:582 a novel approach for nerve repair and remyelination therapies 567:627 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 17 theme differentiation 462:476 arg1 regulation 432:441 regulation 432:441 regulation of oligodendrocyte differentiation and myelination 432:492 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 5 18 theme LRR-Ig-composite 1054:1069 arg1 fold 1071:1074 its LRR-Ig-composite fold 1050:1074 its LRR-Ig-composite fold that drives such assembly 1050:1100 The structure, together with biophysical analysis of its solution properties, reveals that in the crystals and in solution Lingo-1 persistently associates with itself to form a stable tetramer and that it is its LRR-Ig-composite fold that drives such assembly. 17005555 8 19 theme protein 1600:1606 arg1 assembly 1608:1615 protein assembly 1600:1615 protein assembly on the membrane 1600:1631 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 7 20 theme efficient 1340:1348 arg1 scaffold 1350:1357 an efficient scaffold 1337:1357 an efficient scaffold 1337:1357 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 7 20 theme efficient 1340:1348 arg1 tetramer 1272:1279 The tetramer 1268:1279 The tetramer 1268:1279 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 4 21 theme crystal 652:658 arg1 structure 660:668 the crystal structure 648:668 the crystal structure of the ligand-binding ectodomain of human Lingo-1 648:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 0 22 theme repair 87:92 arg1 inhibition 94:103 central nervous system repair inhibition 64:103 central nervous system repair inhibition 64:103 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 8 23 theme binding 1476:1482 arg1 sites 1484:1488 Potential functional binding sites 1455:1488 Potential functional binding sites 1455:1488 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 23 theme binding 1476:1482 arg1 self-recognition 1562:1577 self-recognition 1562:1577 self-recognition 1562:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 8 23 theme binding 1476:1482 arg1 site 1554:1557 the site 1550:1557 the site of self-recognition 1550:1577 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 3 24 theme inhibitory 376:385 arg1 important 419:427 important 419:427 important 419:427 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 24 theme inhibitory 376:385 arg1 role 387:390 the inhibitory role 372:390 the inhibitory role of Lingo-1 372:401 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 6 25 theme crystal 1124:1130 arg1 structure 1132:1140 the crystal structure 1120:1140 the crystal structure 1120:1140 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 1 26 theme axon 146:149 arg1 growth 151:156 axon growth 146:156 axon growth 146:156 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 3 27 from addition 362:369 arg1 important 419:427 important 419:427 important 419:427 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 27 from addition 362:369 arg1 role 387:390 the inhibitory role 372:390 the inhibitory role of Lingo-1 372:401 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 4 28 theme -like 827:831 arg1 modules 833:839 immunoglobulin (Ig)-like modules 808:839 immunoglobulin (Ig)-like modules 808:839 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 5 29 theme properties 908:917 arg1 analysis 883:890 biophysical analysis 871:890 biophysical analysis of its solution properties 871:917 The structure, together with biophysical analysis of its solution properties, reveals that in the crystals and in solution Lingo-1 persistently associates with itself to form a stable tetramer and that it is its LRR-Ig-composite fold that drives such assembly. 17005555 1 30 theme growth 151:156 arg1 control 135:141 Nogo receptor (NgR)-mediated control 106:141 Nogo receptor (NgR)-mediated control of axon growth 106:156 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 4 31 contain has 732:734 arg1 it 729:730 it 729:730 it 729:730 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 4 31 contain has 732:734 arg2 structure 756:764 a bimodular, kinked structure 736:764 structure 756:764 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 3 32 theme novel 569:573 arg1 approach 575:582 a novel approach 567:582 a novel approach for nerve repair and remyelination therapies 567:627 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 32 theme novel 569:573 arg1 modulation 527:536 pharmacological modulation 511:536 pharmacological modulation of Lingo-1 function 511:556 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 33 theme Lingo-1 395:401 arg1 important 419:427 important 419:427 important 419:427 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 33 theme Lingo-1 395:401 arg1 role 387:390 the inhibitory role 372:390 the inhibitory role of Lingo-1 372:401 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 5 34 theme solution 899:906 arg1 properties 908:917 its solution properties 895:917 its solution properties 895:917 The structure, together with biophysical analysis of its solution properties, reveals that in the crystals and in solution Lingo-1 persistently associates with itself to form a stable tetramer and that it is its LRR-Ig-composite fold that drives such assembly. 17005555 4 35 theme bimodular 738:746 arg1 structure 756:764 a bimodular, kinked structure 736:764 structure 756:764 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 5 36 theme biophysical 871:881 arg1 analysis 883:890 biophysical analysis 871:890 biophysical analysis of its solution properties 871:917 The structure, together with biophysical analysis of its solution properties, reveals that in the crystals and in solution Lingo-1 persistently associates with itself to form a stable tetramer and that it is its LRR-Ig-composite fold that drives such assembly. 17005555 4 37 theme ligand-binding 677:690 arg1 ectodomain 692:701 the ligand-binding ectodomain 673:701 the ligand-binding ectodomain of human Lingo-1 673:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 4 37 theme ligand-binding 677:690 arg1 Lingo-1 712:718 human Lingo-1 706:718 human Lingo-1 706:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 0 38 theme ectodomain 29:38 arg1 structure 4:12 The structure 0:12 The structure of the Lingo-1 ectodomain, a module 0:48 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 3 39 theme myelination 482:492 arg1 regulation 432:441 regulation 432:441 regulation of oligodendrocyte differentiation and myelination 432:492 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 40 theme nerve 588:592 arg1 repair 594:599 nerve repair 588:599 nerve repair 588:599 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 1 41 theme central 172:178 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 0 42 theme Lingo-1 21:27 arg1 ectodomain 29:38 the Lingo-1 ectodomain 17:38 the Lingo-1 ectodomain 17:38 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 0 42 theme Lingo-1 21:27 arg1 module 43:48 a module 41:48 a module 41:48 The structure of the Lingo-1 ectodomain, a module implicated in central nervous system repair inhibition. 17005555 4 43 theme human 706:710 arg1 Lingo-1 712:718 human Lingo-1 706:718 human Lingo-1 706:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 3 44 theme repair 594:599 arg1 therapies 619:627 nerve repair and remyelination therapies 588:627 nerve repair and remyelination therapies 588:627 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 45 from important 419:427 arg1 addition 362:369 addition 362:369 addition 362:369 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 45 from important 419:427 arg1 regulation 432:441 regulation 432:441 regulation of oligodendrocyte differentiation and myelination 432:492 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 1 46 theme nervous 180:186 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 4 47 theme Lingo-1 712:718 arg1 ectodomain 692:701 the ligand-binding ectodomain 673:701 the ligand-binding ectodomain of human Lingo-1 673:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 4 47 theme Lingo-1 712:718 arg1 Lingo-1 712:718 human Lingo-1 706:718 human Lingo-1 706:718 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 8 48 theme ectodomain 1520:1529 arg1 surface 1531:1537 the ectodomain surface 1516:1537 the ectodomain surface 1516:1537 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 6 49 theme ring-shaped 1178:1188 arg1 tetramer 1190:1197 a ring-shaped tetramer 1176:1197 a ring-shaped tetramer 1176:1197 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 3 50 from regulation 432:441 arg1 important 419:427 important 419:427 important 419:427 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 50 from regulation 432:441 arg1 role 387:390 the inhibitory role 372:390 the inhibitory role of Lingo-1 372:401 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 1 51 theme Nogo 106:109 arg1 receptor 111:118 Nogo receptor 106:118 Nogo receptor (NgR) 106:124 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 1 51 theme Nogo 106:109 arg1 NgR 121:123 NgR 121:123 NgR 121:123 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 6 52 theme Lingo-1 1155:1161 arg1 protomers 1142:1150 protomers 1142:1150 protomers of Lingo-1 1142:1161 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 6 53 theme adjacent 1249:1256 arg1 protomer 1258:1265 an adjacent protomer 1246:1265 an adjacent protomer 1246:1265 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 4 54 theme leucine-rich 778:789 arg1 repeat 791:796 leucine-rich repeat 778:796 leucine-rich repeat (LRR) 778:802 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 4 54 theme leucine-rich 778:789 arg1 LRR 799:801 LRR 799:801 LRR 799:801 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 3 55 theme oligodendrocyte 446:460 arg1 differentiation 462:476 oligodendrocyte differentiation 446:476 oligodendrocyte differentiation 446:476 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 8 56 from assembly 1608:1615 arg1 membrane 1624:1631 the membrane 1620:1631 the membrane 1620:1631 Potential functional binding sites that can be identified on the ectodomain surface, including the site of self-recognition, suggest a model for protein assembly on the membrane. 17005555 1 57 theme system-specific 188:202 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 5 58 theme stable 1019:1024 arg1 tetramer 1026:1033 a stable tetramer 1017:1033 a stable tetramer 1017:1033 The structure, together with biophysical analysis of its solution properties, reveals that in the crystals and in solution Lingo-1 persistently associates with itself to form a stable tetramer and that it is its LRR-Ig-composite fold that drives such assembly. 17005555 4 59 theme kinked 749:754 arg1 structure 756:764 a bimodular, kinked structure 736:764 structure 756:764 Here we report on the crystal structure of the ligand-binding ectodomain of human Lingo-1 and show it has a bimodular, kinked structure composed of leucine-rich repeat (LRR) and immunoglobulin (Ig)-like modules. 17005555 2 60 theme complementary 293:305 arg1 co-receptor 307:317 a complementary co-receptor 291:317 a complementary co-receptor 291:317 Interactions between Lingo-1 and NgR, along with a complementary co-receptor, result in neurite and axonal collapse. 17005555 1 61 theme type 204:207 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 7 62 theme large 1290:1294 arg1 area 1304:1307 a large surface area 1288:1307 a large surface area (9,200 A2) 1288:1318 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 7 62 theme large 1290:1294 arg1 A2 1316:1317 9,200 A2 1310:1317 9,200 A2 1310:1317 The tetramer buries a large surface area (9,200 A2) and may serve as an efficient scaffold to simultaneously bind and assemble the NgR complex components during activation on a membrane. 17005555 2 63 theme axonal 342:347 arg1 collapse 349:356 axonal collapse 342:356 axonal collapse 342:356 Interactions between Lingo-1 and NgR, along with a complementary co-receptor, result in neurite and axonal collapse. 17005555 6 64 from cleft 1237:1241 arg1 protomer 1258:1265 an adjacent protomer 1246:1265 an adjacent protomer 1246:1265 Specifically, in the crystal structure protomers of Lingo-1 associate in a ring-shaped tetramer, with each LRR domain filling an open cleft in an adjacent protomer. 17005555 1 65 theme I 209:209 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 17005555 3 66 theme remyelination 605:617 arg1 therapies 619:627 nerve repair and remyelination therapies 588:627 nerve repair and remyelination therapies 588:627 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 67 theme pharmacological 511:525 arg1 modulation 527:536 pharmacological modulation 511:536 pharmacological modulation of Lingo-1 function 511:556 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 3 67 theme pharmacological 511:525 arg1 approach 575:582 a novel approach 567:582 a novel approach for nerve repair and remyelination therapies 567:627 In addition, the inhibitory role of Lingo-1 is particularly important in regulation of oligodendrocyte differentiation and myelination, suggesting that pharmacological modulation of Lingo-1 function could be a novel approach for nerve repair and remyelination therapies. 17005555 1 68 theme transmembrane 211:223 arg1 Lingo-1 233:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 the central nervous system-specific type I transmembrane protein Lingo-1 168:239 Nogo receptor (NgR)-mediated control of axon growth relies on the central nervous system-specific type I transmembrane protein Lingo-1. 21525977 5 0 theme sterol 713:718 arg1 pocket 728:733 the sterol binding pocket 709:733 the sterol binding pocket 709:733 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 3 1 attach present 368:374 arg2 we 365:366 we 365:366 we 365:366 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 1 attach present 368:374 arg2 FINDINGS 351:358 PRINCIPAL FINDINGS 341:358 PRINCIPAL FINDINGS 341:358 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 1 attach present 368:374 arg1 absence 452:458 the absence 448:458 the absence of cholesterol 448:473 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 1 2 theme drug 142:145 arg1 Ezetimibe 147:155 the cholesterol lowering drug Ezetimibe 117:155 the cholesterol lowering drug Ezetimibe 117:155 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 6 3 theme multiple 933:940 arg1 movements 942:950 multiple movements 933:950 multiple movements around the entrance to the sterol binding pocket 933:999 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 5 4 theme binding 720:726 arg1 pocket 728:733 the sterol binding pocket 709:733 the sterol binding pocket 709:733 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 0 5 from structure 4:12 arg1 conformation 58:69 a closed conformation 49:69 a closed conformation 49:69 The structure of the NPC1L1 N-terminal domain in a closed conformation. 21525977 5 6 theme NPC1 642:645 arg1 structures 628:637 the cholesterol free and bound structures 597:637 the cholesterol free and bound structures of NPC1(NTD) 597:650 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 1 7 theme Ezetimibe 147:155 arg1 NPC1L1 83:88 BACKGROUND NPC1L1 72:88 BACKGROUND NPC1L1 72:88 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 1 7 theme Ezetimibe 147:155 arg1 target 107:112 the molecular target 93:112 the molecular target of the cholesterol lowering drug Ezetimibe 93:155 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 4 8 theme selectivity 560:570 arg1 mechanism 535:543 the mechanism 531:543 the mechanism of cholesterol selectivity of NPC1L1 531:580 The structure, combined with biochemical data, reveals the mechanism of cholesterol selectivity of NPC1L1. 21525977 2 9 theme cholesterol 321:331 arg1 levels 333:338 plasma cholesterol levels 314:338 plasma cholesterol levels 314:338 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 3 10 theme cholesterol 463:473 arg1 absence 452:458 the absence 448:458 the absence of cholesterol 448:473 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 5 11 theme closed 685:690 arg1 conformation 692:703 a closed conformation 683:703 a closed conformation 683:703 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 6 12 theme flexibility 821:831 arg1 degree 811:816 a degree 809:816 a degree of flexibility surrounding the entrance to the sterol binding pocket 809:885 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 5 13 theme free 613:616 arg1 structures 628:637 the cholesterol free and bound structures 597:637 the cholesterol free and bound structures of NPC1(NTD) 597:650 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 5 14 theme cholesterol 601:611 arg1 structures 628:637 the cholesterol free and bound structures 597:637 the cholesterol free and bound structures of NPC1(NTD) 597:650 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 4 15 theme NPC1L1 575:580 arg1 selectivity 560:570 cholesterol selectivity 548:570 cholesterol selectivity of NPC1L1 548:580 The structure, combined with biochemical data, reveals the mechanism of cholesterol selectivity of NPC1L1. 21525977 3 16 theme N-terminal 411:420 arg1 NTD 430:432 NTD 430:432 NTD 430:432 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 16 theme N-terminal 411:420 arg1 NPC1L1 438:443 NPC1L1 438:443 NPC1L1 438:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 16 theme N-terminal 411:420 arg1 domain 422:427 the N-terminal domain 407:427 the N-terminal domain (NTD) of NPC1L1 407:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 17 theme crystal 386:392 arg1 structure 394:402 the 2.8 Å crystal structure 376:402 the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 376:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 5 18 theme bound 622:626 arg1 structures 628:637 the cholesterol free and bound structures 597:637 the cholesterol free and bound structures of NPC1(NTD) 597:650 Comparison to the cholesterol free and bound structures of NPC1(NTD) reveals that NPC1L1(NTD) is in a closed conformation and the sterol binding pocket is occluded from solvent. 21525977 3 19 theme PRINCIPAL 341:349 arg1 FINDINGS 351:358 PRINCIPAL FINDINGS 341:358 PRINCIPAL FINDINGS 341:358 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 1 20 theme BACKGROUND 72:81 arg1 NPC1L1 83:88 BACKGROUND NPC1L1 72:88 BACKGROUND NPC1L1 72:88 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 1 20 theme BACKGROUND 72:81 arg1 target 107:112 the molecular target 93:112 the molecular target of the cholesterol lowering drug Ezetimibe 93:155 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 2 21 theme plasma 314:319 arg1 levels 333:338 plasma cholesterol levels 314:338 plasma cholesterol levels 314:338 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 1 22 theme intestinal 174:183 arg1 absorption 185:194 the intestinal absorption 170:194 the intestinal absorption of cholesterol 170:209 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 6 23 theme gating 901:906 arg1 mechanism 908:916 a gating mechanism 899:916 a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket 899:999 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 6 24 theme binding 986:992 arg1 pocket 994:999 the sterol binding pocket 975:999 the sterol binding pocket 975:999 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 3 25 theme Å 384:384 arg1 structure 394:402 the 2.8 Å crystal structure 376:402 the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 376:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 4 26 theme cholesterol 548:558 arg1 selectivity 560:570 cholesterol selectivity 548:570 cholesterol selectivity of NPC1L1 548:580 The structure, combined with biochemical data, reveals the mechanism of cholesterol selectivity of NPC1L1. 21525977 6 27 theme sterol 979:984 arg1 pocket 994:999 the sterol binding pocket 975:999 the sterol binding pocket 975:999 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 0 28 theme N-terminal 28:37 arg1 domain 39:44 the NPC1L1 N-terminal domain 17:44 the NPC1L1 N-terminal domain 17:44 The structure of the NPC1L1 N-terminal domain in a closed conformation. 21525977 3 29 theme NPC1L1 438:443 arg1 NTD 430:432 NTD 430:432 NTD 430:432 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 29 theme NPC1L1 438:443 arg1 NPC1L1 438:443 NPC1L1 438:443 NPC1L1 438:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 3 29 theme NPC1L1 438:443 arg1 domain 422:427 the N-terminal domain 407:427 the N-terminal domain (NTD) of NPC1L1 407:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 6 30 dep CONCLUSION 761:770 arg1 reveals 801:807 reveals 801:807 reveals 801:807 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 1 31 theme cholesterol 199:209 arg1 absorption 185:194 the intestinal absorption 170:194 the intestinal absorption of cholesterol 170:209 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 0 32 theme NPC1L1 21:26 arg1 domain 39:44 the NPC1L1 N-terminal domain 17:44 the NPC1L1 N-terminal domain 17:44 The structure of the NPC1L1 N-terminal domain in a closed conformation. 21525977 3 33 theme domain 422:427 arg1 structure 394:402 the 2.8 Å crystal structure 376:402 the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 376:443 PRINCIPAL FINDINGS Here we present the 2.8 Å crystal structure of the N-terminal domain (NTD) of NPC1L1 in the absence of cholesterol. 21525977 4 34 theme biochemical 505:515 arg1 data 517:520 biochemical data 505:520 biochemical data 505:520 The structure, combined with biochemical data, reveals the mechanism of cholesterol selectivity of NPC1L1. 21525977 1 35 theme molecular 97:105 arg1 NPC1L1 83:88 BACKGROUND NPC1L1 72:88 BACKGROUND NPC1L1 72:88 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 1 35 theme molecular 97:105 arg1 target 107:112 the molecular target 93:112 the molecular target of the cholesterol lowering drug Ezetimibe 93:155 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 0 36 theme domain 39:44 arg1 structure 4:12 The structure 0:12 The structure of the NPC1L1 N-terminal domain in a closed conformation 0:69 The structure of the NPC1L1 N-terminal domain in a closed conformation. 21525977 6 37 theme binding 872:878 arg1 pocket 880:885 the sterol binding pocket 861:885 the sterol binding pocket 861:885 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 6 38 theme sterol 865:870 arg1 pocket 880:885 the sterol binding pocket 861:885 the sterol binding pocket 861:885 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 6 39 theme NPC1L1 789:794 arg1 structure 776:784 The structure 772:784 The structure of NPC1L1(NTD) 772:799 CONCLUSION The structure of NPC1L1(NTD) reveals a degree of flexibility surrounding the entrance to the sterol binding pocket, suggesting a gating mechanism that relies on multiple movements around the entrance to the sterol binding pocket. 21525977 2 40 theme cholesterol 264:274 arg1 absorption 276:285 intestinal cholesterol absorption 253:285 intestinal cholesterol absorption 253:285 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 2 41 theme intestinal 253:262 arg1 absorption 276:285 intestinal cholesterol absorption 253:285 intestinal cholesterol absorption 253:285 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 2 42 theme levels 333:338 arg1 reduction 301:309 reduction 301:309 reduction of plasma cholesterol levels 301:338 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 0 43 theme closed 51:56 arg1 conformation 58:69 a closed conformation 49:69 a closed conformation 49:69 The structure of the NPC1L1 N-terminal domain in a closed conformation. 21525977 1 44 theme cholesterol 121:131 arg1 Ezetimibe 147:155 the cholesterol lowering drug Ezetimibe 117:155 the cholesterol lowering drug Ezetimibe 117:155 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21525977 2 45 theme NPC1L1 238:243 arg1 deletion 226:233 deletion 226:233 deletion 226:233 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 2 45 theme NPC1L1 238:243 arg1 Inhibition 212:221 Inhibition 212:221 Inhibition 212:221 Inhibition or deletion of NPC1L1 reduces intestinal cholesterol absorption, resulting in reduction of plasma cholesterol levels. 21525977 1 46 theme lowering 133:140 arg1 Ezetimibe 147:155 the cholesterol lowering drug Ezetimibe 117:155 the cholesterol lowering drug Ezetimibe 117:155 BACKGROUND NPC1L1 is the molecular target of the cholesterol lowering drug Ezetimibe and mediates the intestinal absorption of cholesterol. 21205830 0 0 theme finger 76:81 arg1 proteins 83:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 1 1 theme RING 331:334 arg1 ligases 356:362 membrane-spanning RING finger E3 ubiquitin ligases 313:362 membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 313:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 1 2 theme protein 238:244 arg1 degradation 246:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 0 3 theme RING 71:74 arg1 proteins 83:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 1 4 theme finger 336:341 arg1 ligases 356:362 membrane-spanning RING finger E3 ubiquitin ligases 313:362 membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 313:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 4 5 theme turnover 915:922 arg1 mechanism 893:901 an ER-associated degradation-like mechanism 859:901 an ER-associated degradation-like mechanism of calnexin turnover 859:922 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 1 6 theme degradation 246:256 arg1 regulators 190:199 novel regulators 184:199 novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function 184:272 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 1 7 theme E3 343:344 arg1 ligases 356:362 membrane-spanning RING finger E3 ubiquitin ligases 313:362 membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 313:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 3 8 theme wild 540:543 arg1 Nixin 550:554 wild type Nixin 540:554 wild type Nixin 540:554 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 1 9 theme ubiquitin 346:354 arg1 ligases 356:362 membrane-spanning RING finger E3 ubiquitin ligases 313:362 membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 313:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 3 10 theme type 545:548 arg1 Nixin 550:554 wild type Nixin 540:554 wild type Nixin 540:554 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 1 11 theme ER 262:263 arg1 function 265:272 ER function 262:272 ER function 262:272 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 2 12 theme turnover 508:515 arg1 regulation 485:494 the regulation 481:494 the regulation of calnexin turnover 481:515 We identified 24 ER membrane-anchored ubiquitin ligases and found Nixin/ZNRF4 to be central for the regulation of calnexin turnover. 21205830 1 13 theme ligases 356:362 arg1 inventory 300:308 the entire inventory 289:308 the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 289:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 2 14 theme calnexin 499:506 arg1 turnover 508:515 calnexin turnover 499:515 calnexin turnover 499:515 We identified 24 ER membrane-anchored ubiquitin ligases and found Nixin/ZNRF4 to be central for the regulation of calnexin turnover. 21205830 1 15 theme localized 364:372 arg1 ligases 356:362 membrane-spanning RING finger E3 ubiquitin ligases 313:362 membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 313:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 4 16 theme degradation-like 876:891 arg1 mechanism 893:901 an ER-associated degradation-like mechanism 859:901 an ER-associated degradation-like mechanism of calnexin turnover 859:922 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 4 17 theme calnexin 792:799 arg1 ubiquitination 801:814 calnexin ubiquitination 792:814 calnexin ubiquitination 792:814 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 0 18 theme systematic 2:11 arg1 search 13:18 A systematic search 0:18 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins 0:90 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 4 19 theme calnexin 906:913 arg1 turnover 915:922 calnexin turnover 906:922 calnexin turnover 906:922 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 1 20 link -linked 230:236 arg1 degradation 246:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 1 21 theme function 265:272 arg1 regulators 190:199 novel regulators 184:199 novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function 184:272 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 3 22 theme dramatic 566:573 arg1 down-regulation 575:589 a dramatic down-regulation 564:589 a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain 564:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 1 23 theme novel 184:188 arg1 regulators 190:199 novel regulators 184:199 novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function 184:272 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 3 24 theme Nixin 550:554 arg1 expression 526:535 Ectopic expression 518:535 Ectopic expression of wild type Nixin 518:554 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 2 25 theme membrane-anchored 405:421 arg1 ligases 433:439 24 ER membrane-anchored ubiquitin ligases 399:439 24 ER membrane-anchored ubiquitin ligases 399:439 We identified 24 ER membrane-anchored ubiquitin ligases and found Nixin/ZNRF4 to be central for the regulation of calnexin turnover. 21205830 2 26 theme ubiquitin 423:431 arg1 ligases 433:439 24 ER membrane-anchored ubiquitin ligases 399:439 24 ER membrane-anchored ubiquitin ligases 399:439 We identified 24 ER membrane-anchored ubiquitin ligases and found Nixin/ZNRF4 to be central for the regulation of calnexin turnover. 21205830 4 27 theme glycosylation-independent 750:774 arg1 manner 776:781 a glycosylation-independent manner 748:781 a glycosylation-independent manner 748:781 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 2 28 theme ER 402:403 arg1 ligases 433:439 24 ER membrane-anchored ubiquitin ligases 399:439 24 ER membrane-anchored ubiquitin ligases 399:439 We identified 24 ER membrane-anchored ubiquitin ligases and found Nixin/ZNRF4 to be central for the regulation of calnexin turnover. 21205830 0 29 theme reticulum 36:44 arg1 proteins 83:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 0 30 theme calnexin 133:140 arg1 stability 142:150 calnexin stability 133:150 calnexin stability 133:150 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 1 31 theme endoplasmic 204:214 arg1 ER 227:228 ER 227:228 ER 227:228 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 1 31 theme endoplasmic 204:214 arg1 reticulum 216:224 endoplasmic reticulum 204:224 endoplasmic reticulum (ER)-linked protein degradation 204:256 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 0 32 theme endoplasmic 24:34 arg1 reticulum 36:44 endoplasmic reticulum 24:44 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 0 32 theme endoplasmic 24:34 arg1 ER 47:48 ER 47:48 ER 47:48 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 3 33 theme ER-localized 598:609 arg1 calnexin 621:628 the ER-localized chaperone calnexin 594:628 the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain 594:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 1 34 theme reticulum 216:224 arg1 degradation 246:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 1 35 theme entire 293:298 arg1 inventory 300:308 the entire inventory 289:308 the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 289:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 4 36 theme ER-associated 862:874 arg1 mechanism 893:901 an ER-associated degradation-like mechanism 859:901 an ER-associated degradation-like mechanism of calnexin turnover 859:922 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 0 37 theme stability 142:150 arg1 Nixin/ZNRF4 103:113 Nixin/ZNRF4 103:113 Nixin/ZNRF4 103:113 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 0 37 theme stability 142:150 arg1 regulator 120:128 a regulator 118:128 a regulator of calnexin stability and ER homeostasis 118:169 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 3 38 theme RING 678:681 arg1 domain 683:688 the Nixin RING domain 668:688 the Nixin RING domain 668:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 4 39 theme p97-dependent 821:833 arg1 degradation 835:845 p97-dependent degradation 821:845 p97-dependent degradation 821:845 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 0 40 theme homeostasis 159:169 arg1 Nixin/ZNRF4 103:113 Nixin/ZNRF4 103:113 Nixin/ZNRF4 103:113 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 0 40 theme homeostasis 159:169 arg1 regulator 120:128 a regulator 118:128 a regulator of calnexin stability and ER homeostasis 118:169 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 3 41 theme domain 683:688 arg1 inactivation 652:663 inactivation 652:663 inactivation of the Nixin RING domain 652:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 4 42 dep interacts 721:729 arg1 induces 784:790 induces 784:790 induces calnexin ubiquitination, and p97-dependent degradation 784:845 Importantly, Nixin physically interacts with calnexin in a glycosylation-independent manner, induces calnexin ubiquitination, and p97-dependent degradation, indicating an ER-associated degradation-like mechanism of calnexin turnover. 21205830 0 43 theme ER 156:157 arg1 homeostasis 159:169 ER homeostasis 156:169 ER homeostasis 156:169 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 0 44 theme membrane-associated 51:69 arg1 proteins 83:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 endoplasmic reticulum (ER) membrane-associated RING finger proteins 24:90 A systematic search for endoplasmic reticulum (ER) membrane-associated RING finger proteins identifies Nixin/ZNRF4 as a regulator of calnexin stability and ER homeostasis. 21205830 3 45 theme Nixin 672:676 arg1 domain 683:688 the Nixin RING domain 668:688 the Nixin RING domain 668:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 3 46 theme chaperone 611:619 arg1 calnexin 621:628 the ER-localized chaperone calnexin 594:628 the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain 594:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 1 47 theme membrane-spanning 313:329 arg1 ligases 356:362 membrane-spanning RING finger E3 ubiquitin ligases 313:362 membrane-spanning RING finger E3 ubiquitin ligases localized to the ER 313:382 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 21205830 3 48 theme calnexin 621:628 arg1 down-regulation 575:589 a dramatic down-regulation 564:589 a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain 564:688 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 3 49 theme Ectopic 518:524 arg1 expression 526:535 Ectopic expression 518:535 Ectopic expression of wild type Nixin 518:554 Ectopic expression of wild type Nixin induced a dramatic down-regulation of the ER-localized chaperone calnexin that was prevented by inactivation of the Nixin RING domain. 21205830 1 50 theme -linked 230:236 arg1 degradation 246:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 endoplasmic reticulum (ER)-linked protein degradation 204:256 To identify novel regulators of endoplasmic reticulum (ER)-linked protein degradation and ER function, we determined the entire inventory of membrane-spanning RING finger E3 ubiquitin ligases localized to the ER. 19838198 5 0 from receptor 940:947 arg1 complexes 981:989 multiple different signaling complexes 952:989 multiple different signaling complexes 952:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 1 1 theme H 117:117 arg1 cells 108:112 Interleukin 17 (IL-17)-producing helper T cells 66:112 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells) 66:128 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 1 theme H 117:117 arg1 cells 123:127 T(H)-17 cells 115:127 T(H)-17 cells 115:127 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 2 theme autoimmune 268:277 arg1 disorders 279:287 chronic inflammatory and autoimmune disorders 243:287 chronic inflammatory and autoimmune disorders 243:287 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 4 3 theme complex 783:789 arg1 formation 791:799 heterodimeric versus homodimeric complex formation 750:799 heterodimeric versus homodimeric complex formation 750:799 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 5 4 theme common 817:822 arg1 strategy 836:843 a common recognition strategy 815:843 a common recognition strategy 815:843 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 5 5 theme recognition 824:834 arg1 strategy 836:843 a common recognition strategy 815:843 a common recognition strategy 815:843 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 1 6 theme disorders 279:287 arg1 mediators 230:238 key mediators 226:238 key mediators of chronic inflammatory and autoimmune disorders 226:287 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 5 7 theme IL-17 879:883 arg1 family 885:890 the IL-17 family 875:890 the IL-17 family 875:890 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 4 8 theme IL-17 639:643 arg1 cytokines 645:653 the IL-17 cytokines 635:653 the IL-17 cytokines 635:653 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 1 9 theme IL-17 196:200 arg1 family 202:207 the IL-17 family 192:207 the IL-17 family 192:207 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 4 10 theme receptor-binding 708:723 arg1 event 725:729 the second receptor-binding event 697:729 the second receptor-binding event 697:729 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 3 11 theme homodimer 581:589 arg1 interface 591:599 the IL-17 homodimer interface 571:599 the IL-17 homodimer interface 571:599 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 5 12 theme family 885:890 arg1 members 864:870 several members 856:870 several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes 856:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 1 13 theme family 202:207 arg1 members 181:187 members 181:187 members of the IL-17 family 181:207 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 4 14 theme heterodimeric 750:762 arg1 formation 791:799 heterodimeric versus homodimeric complex formation 750:799 heterodimeric versus homodimeric complex formation 750:799 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 3 15 theme complex 428:434 arg1 formation 436:444 complex formation 428:444 complex formation 428:444 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 1 16 theme -17 119:121 arg1 cells 108:112 Interleukin 17 (IL-17)-producing helper T cells 66:112 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells) 66:128 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 16 theme -17 119:121 arg1 cells 123:127 T(H)-17 cells 115:127 T(H)-17 cells 115:127 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 3 17 theme formation 436:444 arg1 unique 450:455 unique 450:455 unique 450:455 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 3 17 theme formation 436:444 arg1 mechanism 415:423 The mechanism 411:423 The mechanism of complex formation 411:444 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 4 18 theme homodimeric 771:781 arg1 formation 791:799 heterodimeric versus homodimeric complex formation 750:799 heterodimeric versus homodimeric complex formation 750:799 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 5 19 theme several 856:862 arg1 members 864:870 several members 856:870 several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes 856:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 2 20 theme crystal 310:316 arg1 structure 318:326 the crystal structure 306:326 the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry 306:408 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 0 21 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of receptor 0:27 Structural basis of receptor sharing by interleukin 17 cytokines. 19838198 0 22 theme receptor 20:27 arg1 basis 11:15 Structural basis 0:15 Structural basis of receptor 0:27 Structural basis of receptor sharing by interleukin 17 cytokines. 19838198 4 23 theme second 701:706 arg1 event 725:729 the second receptor-binding event 697:729 the second receptor-binding event 697:729 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 5 24 theme shared 933:938 arg1 receptor 940:947 a shared receptor 931:947 a shared receptor in multiple different signaling complexes 931:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 5 24 theme shared 933:938 arg1 it 906:907 it 906:907 it 906:907 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 2 25 theme IL-17 344:348 arg1 IL-17RA 362:368 IL-17RA 362:368 IL-17RA 362:368 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 2 25 theme IL-17 344:348 arg1 A 359:359 IL-17 receptor A 344:359 IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry 344:408 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 2 26 theme receptor 350:357 arg1 IL-17RA 362:368 IL-17RA 362:368 IL-17RA 362:368 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 2 26 theme receptor 350:357 arg1 A 359:359 IL-17 receptor A 344:359 IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry 344:408 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 5 27 theme signaling 971:979 arg1 complexes 981:989 multiple different signaling complexes 952:989 multiple different signaling complexes 952:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 1 28 theme -producing 88:97 arg1 cells 108:112 Interleukin 17 (IL-17)-producing helper T cells 66:112 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells) 66:128 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 28 theme -producing 88:97 arg1 cells 123:127 T(H)-17 cells 115:127 T(H)-17 cells 115:127 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 4 29 theme event 725:729 arg1 specificity 682:692 specificity 682:692 specificity 682:692 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 4 29 theme event 725:729 arg1 affinity 669:676 affinity 669:676 affinity 669:676 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 2 30 theme complex 333:339 arg1 structure 318:326 the crystal structure 306:326 the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry 306:408 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 1 31 theme key 226:228 arg1 mediators 230:238 key mediators 226:238 key mediators of chronic inflammatory and autoimmune disorders 226:287 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 2 32 theme A 359:359 arg1 complex 333:339 a complex 331:339 a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry 331:408 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 1 33 theme effector 151:158 arg1 cytokines 160:168 their effector cytokines 145:168 their effector cytokines 145:168 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 3 34 theme IL-17RA 543:549 arg1 IL-17RA 543:549 IL-17RA 543:549 IL-17RA 543:549 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 3 34 theme IL-17RA 543:549 arg1 domains 532:538 two fibronectin-type domains 511:538 two fibronectin-type domains of IL-17RA 511:549 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 1 35 theme helper 99:104 arg1 cells 108:112 Interleukin 17 (IL-17)-producing helper T cells 66:112 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells) 66:128 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 35 theme helper 99:104 arg1 cells 123:127 T(H)-17 cells 115:127 T(H)-17 cells 115:127 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 0 36 theme interleukin 40:50 arg1 cytokines 55:63 interleukin 17 cytokines 40:63 interleukin 17 cytokines 40:63 Structural basis of receptor sharing by interleukin 17 cytokines. 19838198 5 37 used used 810:813 arg2 IL-17RA 802:808 IL-17RA 802:808 IL-17RA 802:808 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 1 38 theme T 106:106 arg1 cells 108:112 Interleukin 17 (IL-17)-producing helper T cells 66:112 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells) 66:128 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 38 theme T 106:106 arg1 cells 123:127 T(H)-17 cells 115:127 T(H)-17 cells 115:127 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 39 theme chronic 243:249 arg1 disorders 279:287 chronic inflammatory and autoimmune disorders 243:287 chronic inflammatory and autoimmune disorders 243:287 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 3 40 theme IL-17 575:579 arg1 interface 591:599 the IL-17 homodimer interface 571:599 the IL-17 homodimer interface 571:599 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 3 41 theme fibronectin-type 515:530 arg1 IL-17RA 543:549 IL-17RA 543:549 IL-17RA 543:549 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 3 41 theme fibronectin-type 515:530 arg1 domains 532:538 two fibronectin-type domains 511:538 two fibronectin-type domains of IL-17RA 511:549 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 3 42 theme IL-17 502:506 arg1 engagement 488:497 the engagement 484:497 the engagement of IL-17 by two fibronectin-type domains of IL-17RA 484:549 The mechanism of complex formation was unique for cytokines and involved the engagement of IL-17 by two fibronectin-type domains of IL-17RA in a groove between the IL-17 homodimer interface. 19838198 4 43 theme receptor 623:630 arg1 Binding 602:608 Binding 602:608 Binding of the first receptor to the IL-17 cytokines 602:653 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 5 44 theme multiple 952:959 arg1 complexes 981:989 multiple different signaling complexes 952:989 multiple different signaling complexes 952:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 4 45 theme first 617:621 arg1 receptor 623:630 the first receptor 613:630 the first receptor 613:630 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 19838198 5 46 theme different 961:969 arg1 complexes 981:989 multiple different signaling complexes 952:989 multiple different signaling complexes 952:989 IL-17RA used a common recognition strategy to bind to several members of the IL-17 family, which allows it to potentially act as a shared receptor in multiple different signaling complexes. 19838198 1 47 dep together 131:138 arg1 with 140:143 with 140:143 with 140:143 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 48 theme T 115:115 arg1 cells 108:112 Interleukin 17 (IL-17)-producing helper T cells 66:112 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells) 66:128 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 1 48 theme T 115:115 arg1 cells 123:127 T(H)-17 cells 115:127 T(H)-17 cells 115:127 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 2 49 theme 1:2 392:394 arg1 stoichiometry 396:408 a 1:2 stoichiometry 390:408 a 1:2 stoichiometry 390:408 Here we present the crystal structure of a complex of IL-17 receptor A (IL-17RA) bound to IL-17F in a 1:2 stoichiometry. 19838198 1 50 theme inflammatory 251:262 arg1 disorders 279:287 chronic inflammatory and autoimmune disorders 243:287 chronic inflammatory and autoimmune disorders 243:287 Interleukin 17 (IL-17)-producing helper T cells (T(H)-17 cells), together with their effector cytokines, including members of the IL-17 family, are emerging as key mediators of chronic inflammatory and autoimmune disorders. 19838198 4 51 dep affinity 669:676 arg1 the 665:667 the 665:667 the 665:667 Binding of the first receptor to the IL-17 cytokines modulated the affinity and specificity of the second receptor-binding event, thereby promoting heterodimeric versus homodimeric complex formation. 31959827 4 0 theme better 574:579 arg1 approaches 581:590 better approaches 574:590 better approaches 574:590 The clustered and variable O-glycans make the IgA1 glycomic analysis challenging and better approaches are needed. 31959827 6 1 with profiles 762:769 arg1 deglycosylation 787:801 sequential deglycosylation 776:801 sequential deglycosylation 776:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 10 2 theme quantitative 1261:1272 arg1 assignment 1274:1283 quantitative assignment 1261:1283 quantitative assignment of Gd sites 1261:1295 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 31959827 3 3 with glycoforms 392:401 arg1 O-glycans 441:449 some galactose-deficient (Gd) HR O-glycans 408:449 some galactose-deficient (Gd) HR O-glycans 408:449 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 2 4 link β1,3-linked 339:349 arg1 galactose 351:359 β1,3-linked galactose 339:359 β1,3-linked galactose 339:359 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 5 5 theme IgA1 660:663 arg1 analysis 680:687 IgA1 HR O-glycoform analysis 660:687 IgA1 HR O-glycoform analysis 660:687 Here, we report a comprehensive analytical workflow for IgA1 HR O-glycoform analysis. 31959827 10 6 theme glycoforms 1242:1251 arg1 chromatograms 1216:1228 Extracted ion chromatograms 1202:1228 Extracted ion chromatograms of isomeric glycoforms 1202:1251 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 31959827 12 7 theme O-glycoforms 1429:1440 arg1 profiling 1408:1416 quantitative profiling 1395:1416 quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution 1395:1470 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 8 theme HR 1426:1427 arg1 O-glycoforms 1429:1440 IgA1 HR O-glycoforms 1421:1440 IgA1 HR O-glycoforms 1421:1440 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 8 9 dep three 1044:1048 arg1 to 1041:1042 to 1041:1042 to 1041:1042 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 5 10 theme comprehensive 622:634 arg1 workflow 647:654 a comprehensive analytical workflow 620:654 a comprehensive analytical workflow for IgA1 HR O-glycoform analysis 620:687 Here, we report a comprehensive analytical workflow for IgA1 HR O-glycoform analysis. 31959827 2 11 theme clustered 263:271 arg1 O-glycans 273:281 up to six clustered O-glycans 253:281 up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation 253:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 12 12 from IgAN 1537:1540 arg1 identification 1484:1497 identification 1484:1497 identification of pathogenic IgA1 HR O-glycoforms in IgAN 1484:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 13 theme quantitative 1395:1406 arg1 profiling 1408:1416 quantitative profiling 1395:1416 quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution 1395:1470 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 6 14 dep all 813:815 arg1 O-glycans 824:832 O-glycans 824:832 O-glycans 824:832 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 12 15 from O-glycoforms 1521:1532 arg1 IgAN 1537:1540 IgAN 1537:1540 IgAN 1537:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 6 16 gly deglycosylation 787:801 arg1 profiles 762:769 the HR O-glycopeptide profiles 740:769 the HR O-glycopeptide profiles with sequential deglycosylation 740:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 3 17 theme Gd 434:435 arg1 O-glycans 441:449 some galactose-deficient (Gd) HR O-glycans 408:449 some galactose-deficient (Gd) HR O-glycans 408:449 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 2 18 dep six 259:261 arg1 to 256:257 to 256:257 to 256:257 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 5 19 theme O-glycoform 668:678 arg1 analysis 680:687 IgA1 HR O-glycoform analysis 660:687 IgA1 HR O-glycoform analysis 660:687 Here, we report a comprehensive analytical workflow for IgA1 HR O-glycoform analysis. 31959827 1 20 theme immunoglobulin 105:118 arg1 A 120:120 immunoglobulin A 105:120 immunoglobulin A (IgA) nephropathy (IgAN) 105:145 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 1 20 theme immunoglobulin 105:118 arg1 IgA 123:125 IgA 123:125 IgA 123:125 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 10 21 theme sites 1291:1295 arg1 assignment 1274:1283 quantitative assignment 1261:1283 quantitative assignment of Gd sites 1261:1295 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 31959827 9 22 with Sites 1064:1068 arg1 O-glycans 1078:1086 Gd O-glycans 1075:1086 Gd O-glycans 1075:1086 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 2 23 theme IgA1 226:229 arg1 HR 245:246 HR 245:246 HR 245:246 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 2 23 theme IgA1 226:229 arg1 region 237:242 IgA1 hinge region 226:242 IgA1 hinge region (HR) 226:247 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 4 24 theme clustered 493:501 arg1 O-glycans 516:524 The clustered and variable O-glycans 489:524 The clustered and variable O-glycans 489:524 The clustered and variable O-glycans make the IgA1 glycomic analysis challenging and better approaches are needed. 31959827 3 25 gly glycoforms 392:401 arg1 IgA1 387:390 IgA1 glycoforms 387:401 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans 387:449 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 6 26 gly O-glycopeptide 747:760 arg2 O-glycopeptide 747:760 the HR O-glycopeptide profiles 740:769 the HR O-glycopeptide profiles with sequential deglycosylation 740:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 1 27 theme common 83:88 arg1 nephropathy 128:138 immunoglobulin A (IgA) nephropathy 105:138 immunoglobulin A (IgA) nephropathy (IgAN) 105:145 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 1 27 theme common 83:88 arg1 disease 96:102 A common renal disease 81:102 A common renal disease 81:102 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 4 28 theme IgA1 535:538 arg1 analysis 549:556 the IgA1 glycomic analysis 531:556 the IgA1 glycomic analysis 531:556 The clustered and variable O-glycans make the IgA1 glycomic analysis challenging and better approaches are needed. 31959827 8 29 dep six 982:984 arg1 to 979:980 to 979:980 to 979:980 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 0 30 theme O-glycoforms 12:23 arg1 Analysis 0:7 Analysis 0:7 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 0:79 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 31959827 2 31 contain has 249:251 arg2 O-glycans 273:281 up to six clustered O-glycans 253:281 up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation 253:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 2 31 contain has 249:251 arg1 HR 245:246 HR 245:246 HR 245:246 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 2 31 contain has 249:251 arg1 region 237:242 IgA1 hinge region 226:242 IgA1 hinge region (HR) 226:247 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 10 32 theme Extracted 1202:1210 arg1 chromatograms 1216:1228 Extracted ion chromatograms 1202:1228 Extracted ion chromatograms of isomeric glycoforms 1202:1251 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 31959827 2 33 with N-acetylgalactosamine 312:332 arg1 galactose 351:359 β1,3-linked galactose 339:359 β1,3-linked galactose 339:359 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 2 33 with N-acetylgalactosamine 312:332 arg1 sialylation 374:384 variable sialylation 365:384 variable sialylation 365:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 12 34 theme site-specific 1447:1459 arg1 resolution 1461:1470 site-specific resolution 1447:1470 site-specific resolution 1447:1470 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 4 35 theme variable 507:514 arg1 O-glycans 516:524 The clustered and variable O-glycans 489:524 The clustered and variable O-glycans 489:524 The clustered and variable O-glycans make the IgA1 glycomic analysis challenging and better approaches are needed. 31959827 11 36 theme frequent 1307:1314 arg1 T236 1328:1331 T236 1328:1331 T236 1328:1331 The most frequent Gd site was T236, followed by S230, T233, T228, and S232. 31959827 11 36 theme frequent 1307:1314 arg1 site 1319:1322 The most frequent Gd site 1298:1322 The most frequent Gd site 1298:1322 The most frequent Gd site was T236, followed by S230, T233, T228, and S232. 31959827 6 37 theme profiles 762:769 arg1 analysis 728:735 an automated quantitative analysis 702:735 an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation 702:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 3 38 theme IgAN 470:473 arg1 pathogenesis 475:486 IgAN pathogenesis 470:486 IgAN pathogenesis 470:486 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 0 39 theme hinge 37:41 arg1 region 43:48 the IgA1 hinge region 28:48 the IgA1 hinge region 28:48 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 31959827 12 40 theme IgA1 1513:1516 arg1 O-glycoforms 1521:1532 pathogenic IgA1 HR O-glycoforms 1502:1532 pathogenic IgA1 HR O-glycoforms in IgAN 1502:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 9 41 theme tandem 1176:1181 arg1 spectrometry 1188:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 10 42 theme isomeric 1233:1240 arg1 glycoforms 1242:1251 isomeric glycoforms 1233:1251 isomeric glycoforms 1233:1251 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 31959827 12 43 theme O-glycoforms 1521:1532 arg1 identification 1484:1497 identification 1484:1497 identification of pathogenic IgA1 HR O-glycoforms in IgAN 1484:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 44 gly O-glycoforms 1521:1532 arg1 HR 1518:1519 pathogenic IgA1 HR O-glycoforms 1502:1532 pathogenic IgA1 HR O-glycoforms in IgAN 1502:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 44 gly O-glycoforms 1521:1532 arg2 IgAN 1537:1540 IgAN 1537:1540 IgAN 1537:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 44 gly O-glycoforms 1521:1532 arg1 IgA1 1513:1516 pathogenic IgA1 HR O-glycoforms 1502:1532 pathogenic IgA1 HR O-glycoforms in IgAN 1502:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 45 gly O-glycoforms 1429:1440 arg1 IgA1 1421:1424 IgA1 HR O-glycoforms 1421:1440 IgA1 HR O-glycoforms 1421:1440 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 45 gly O-glycoforms 1429:1440 arg1 HR 1426:1427 IgA1 HR O-glycoforms 1421:1440 IgA1 HR O-glycoforms 1421:1440 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 1 46 theme complexes 215:223 arg1 deposition 178:187 glomerular deposition 167:187 glomerular deposition of IgA1-containing immune complexes 167:223 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 12 47 from identification 1484:1497 arg1 IgAN 1537:1540 IgAN 1537:1540 IgAN 1537:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 8 48 gly glycopeptides 930:942 arg2 glycopeptides 930:942 glycopeptides 930:942 glycopeptides corresponding to the HR with three to six O-glycans 930:994 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 5 49 theme analytical 636:645 arg1 workflow 647:654 a comprehensive analytical workflow 620:654 a comprehensive analytical workflow for IgA1 HR O-glycoform analysis 620:687 Here, we report a comprehensive analytical workflow for IgA1 HR O-glycoform analysis. 31959827 6 50 theme quantitative 715:726 arg1 analysis 728:735 an automated quantitative analysis 702:735 an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation 702:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 2 51 theme β1,3-linked 339:349 arg1 galactose 351:359 β1,3-linked galactose 339:359 β1,3-linked galactose 339:359 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 2 52 link Ser/Thr-linked 297:310 arg1 N-acetylgalactosamine 312:332 Ser/Thr-linked N-acetylgalactosamine 297:332 Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation 297:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 7 53 theme serum 877:881 arg1 IgA1 883:886 serum IgA1 877:886 serum IgA1 from healthy subjects 877:908 The workflow was tested using serum IgA1 from healthy subjects. 31959827 9 54 theme collision 1153:1161 arg1 spectrometry 1188:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 12 55 theme IgA1 1421:1424 arg1 O-glycoforms 1429:1440 IgA1 HR O-glycoforms 1421:1440 IgA1 HR O-glycoforms 1421:1440 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 56 theme new 1378:1380 arg1 workflow 1382:1389 The new workflow 1374:1389 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution 1374:1470 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 6 57 theme sequential 776:785 arg1 deglycosylation 787:801 sequential deglycosylation 776:801 sequential deglycosylation 776:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 6 58 theme O-glycopeptide 747:760 arg1 profiles 762:769 the HR O-glycopeptide profiles 740:769 the HR O-glycopeptide profiles with sequential deglycosylation 740:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 1 59 theme glomerular 167:176 arg1 deposition 178:187 glomerular deposition 167:187 glomerular deposition of IgA1-containing immune complexes 167:223 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 5 60 theme HR 665:666 arg1 analysis 680:687 IgA1 HR O-glycoform analysis 660:687 IgA1 HR O-glycoform analysis 660:687 Here, we report a comprehensive analytical workflow for IgA1 HR O-glycoform analysis. 31959827 8 61 gly glycopeptides 1016:1028 arg2 glycopeptides 1016:1028 nine glycopeptides 1011:1028 nine glycopeptides 1011:1028 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 1 62 theme A 120:120 arg1 IgAN 141:144 IgAN 141:144 IgAN 141:144 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 1 62 theme A 120:120 arg1 nephropathy 128:138 immunoglobulin A (IgA) nephropathy 105:138 immunoglobulin A (IgA) nephropathy (IgAN) 105:145 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 1 62 theme A 120:120 arg1 disease 96:102 A common renal disease 81:102 A common renal disease 81:102 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 6 63 theme HR 744:745 arg1 profiles 762:769 the HR O-glycopeptide profiles 740:769 the HR O-glycopeptide profiles with sequential deglycosylation 740:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 10 64 theme Gd 1288:1289 arg1 sites 1291:1295 Gd sites 1288:1295 Gd sites 1288:1295 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 31959827 0 65 theme sequential 53:62 arg1 deglycosylation 64:78 sequential deglycosylation 53:78 sequential deglycosylation 53:78 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 31959827 2 66 theme hinge 231:235 arg1 HR 245:246 HR 245:246 HR 245:246 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 2 66 theme hinge 231:235 arg1 region 237:242 IgA1 hinge region 226:242 IgA1 hinge region (HR) 226:247 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 6 67 theme automated 705:713 arg1 analysis 728:735 an automated quantitative analysis 702:735 an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation 702:801 We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. 31959827 8 68 theme Gd 1050:1051 arg1 O-glycans 1053:1061 up to three Gd O-glycans 1038:1061 up to three Gd O-glycans 1038:1061 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 9 69 theme Gd 1075:1076 arg1 O-glycans 1078:1086 Gd O-glycans 1075:1086 Gd O-glycans 1075:1086 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 3 70 theme HR 438:439 arg1 O-glycans 441:449 some galactose-deficient (Gd) HR O-glycans 408:449 some galactose-deficient (Gd) HR O-glycans 408:449 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 3 71 theme IgA1 387:390 arg1 glycoforms 392:401 IgA1 glycoforms 387:401 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans 387:449 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 4 72 theme glycomic 540:547 arg1 analysis 549:556 the IgA1 glycomic analysis 531:556 the IgA1 glycomic analysis 531:556 The clustered and variable O-glycans make the IgA1 glycomic analysis challenging and better approaches are needed. 31959827 2 73 gly sialylation 374:384 arg1 N-acetylgalactosamine 312:332 Ser/Thr-linked N-acetylgalactosamine 297:332 Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation 297:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 7 74 from subjects 901:908 arg1 IgA1 883:886 serum IgA1 877:886 serum IgA1 from healthy subjects 877:908 The workflow was tested using serum IgA1 from healthy subjects. 31959827 2 75 theme variable 365:372 arg1 sialylation 374:384 variable sialylation 365:384 variable sialylation 365:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 1 76 theme renal 90:94 arg1 nephropathy 128:138 immunoglobulin A (IgA) nephropathy 105:138 immunoglobulin A (IgA) nephropathy (IgAN) 105:145 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 1 76 theme renal 90:94 arg1 disease 96:102 A common renal disease 81:102 A common renal disease 81:102 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 3 77 theme key 458:460 arg1 role 462:465 a key role 456:465 a key role 456:465 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 0 78 theme IgA1 32:35 arg1 region 43:48 the IgA1 hinge region 28:48 the IgA1 hinge region 28:48 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 31959827 3 79 theme galactose-deficient 413:431 arg1 O-glycans 441:449 some galactose-deficient (Gd) HR O-glycans 408:449 some galactose-deficient (Gd) HR O-glycans 408:449 IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. 31959827 9 80 theme electron-transfer/higher-energy 1121:1151 arg1 spectrometry 1188:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 8 81 contain carried 1030:1036 arg1 glycopeptides 1016:1028 nine glycopeptides 1011:1028 nine glycopeptides 1011:1028 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 8 81 contain carried 1030:1036 arg2 O-glycans 1053:1061 up to three Gd O-glycans 1038:1061 up to three Gd O-glycans 1038:1061 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 0 82 theme region 43:48 arg1 O-glycoforms 12:23 O-glycoforms 12:23 O-glycoforms of the IgA1 hinge region 12:48 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 31959827 0 83 gly O-glycoforms 12:23 arg1 region 43:48 the IgA1 hinge region 28:48 the IgA1 hinge region 28:48 Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation. 31959827 11 84 theme Gd 1316:1317 arg1 T236 1328:1331 T236 1328:1331 T236 1328:1331 The most frequent Gd site was T236, followed by S230, T233, T228, and S232. 31959827 11 84 theme Gd 1316:1317 arg1 site 1319:1322 The most frequent Gd site 1298:1322 The most frequent Gd site 1298:1322 The most frequent Gd site was T236, followed by S230, T233, T228, and S232. 31959827 9 85 theme dissociation 1163:1174 arg1 spectrometry 1188:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 2 86 theme Ser/Thr-linked 297:310 arg1 N-acetylgalactosamine 312:332 Ser/Thr-linked N-acetylgalactosamine 297:332 Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation 297:384 IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. 31959827 12 87 theme pathogenic 1502:1511 arg1 O-glycoforms 1521:1532 pathogenic IgA1 HR O-glycoforms 1502:1532 pathogenic IgA1 HR O-glycoforms in IgAN 1502:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 8 88 theme glycopeptides 930:942 arg1 variants 918:925 Twelve variants 911:925 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans 911:994 Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. 31959827 9 89 theme mass 1183:1186 arg1 spectrometry 1188:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 electron-transfer/higher-energy collision dissociation tandem mass spectrometry 1121:1199 Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. 31959827 7 90 theme healthy 893:899 arg1 subjects 901:908 healthy subjects 893:908 healthy subjects 893:908 The workflow was tested using serum IgA1 from healthy subjects. 31959827 1 91 theme IgA1-containing immune 192:213 arg1 complexes 215:223 IgA1-containing immune complexes 192:223 IgA1-containing immune complexes 192:223 A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. 31959827 12 92 theme HR 1518:1519 arg1 O-glycoforms 1521:1532 pathogenic IgA1 HR O-glycoforms 1502:1532 pathogenic IgA1 HR O-glycoforms in IgAN 1502:1540 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 12 93 with profiling 1408:1416 arg1 resolution 1461:1470 site-specific resolution 1447:1470 site-specific resolution 1447:1470 The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN. 31959827 10 94 theme ion 1212:1214 arg1 chromatograms 1216:1228 Extracted ion chromatograms 1202:1228 Extracted ion chromatograms of isomeric glycoforms 1202:1251 Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. 20145116 8 0 theme peptide 1159:1165 arg1 loops 1167:1171 three peptide loops 1153:1171 three peptide loops 1153:1171 In the VEGFs, receptor specificity is determined by an N-terminal alpha helix and three peptide loops. 20145116 9 1 from loops 1212:1216 arg1 VEGF-C 1221:1226 VEGF-C 1221:1226 VEGF-C 1221:1226 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 13 2 theme structural 1631:1640 arg1 data 1642:1645 our structural data 1627:1645 our structural data 1627:1645 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 12 3 theme chimeric 1529:1536 arg1 proteins 1538:1545 VEGFR-1/VEGFR-2 chimeric proteins 1513:1545 VEGFR-1/VEGFR-2 chimeric proteins 1513:1545 We therefore created VEGFR-1/VEGFR-2 chimeric proteins to further study receptor specificity. 20145116 5 4 theme predominant 687:697 arg1 VEGFR-2 674:680 VEGFR-2 674:680 VEGFR-2 674:680 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 5 4 theme predominant 687:697 arg1 transducer 699:708 the predominant transducer 683:708 the predominant transducer of signals required for physiological and pathological angiogenesis 683:776 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 9 5 theme loops 1212:1216 arg1 two 1199:1201 two 1199:1201 two 1199:1201 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 9 5 theme loops 1212:1216 arg1 loops 1212:1216 these loops 1206:1216 these loops in VEGF-C 1206:1226 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 1 6 theme lymph 148:152 arg1 formation 161:169 blood and lymph vessel formation 138:169 blood and lymph vessel formation 138:169 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 12 7 theme VEGFR-1/VEGFR-2 1513:1527 arg1 proteins 1538:1545 VEGFR-1/VEGFR-2 chimeric proteins 1513:1545 VEGFR-1/VEGFR-2 chimeric proteins 1513:1545 We therefore created VEGFR-1/VEGFR-2 chimeric proteins to further study receptor specificity. 20145116 9 8 theme VEGFR-2 1236:1242 arg1 subdomains 1244:1253 VEGFR-2 subdomains D2 and D3 1236:1263 VEGFR-2 subdomains D2 and D3 1236:1263 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 9 8 theme VEGFR-2 1236:1242 arg1 D3 1262:1263 D3 1262:1263 D3 1262:1263 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 9 8 theme VEGFR-2 1236:1242 arg1 D2 1255:1256 D2 1255:1256 D2 1255:1256 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 8 9 theme receptor 1085:1092 arg1 specificity 1094:1104 receptor specificity 1085:1104 receptor specificity 1085:1104 In the VEGFs, receptor specificity is determined by an N-terminal alpha helix and three peptide loops. 20145116 7 10 theme twisted 1007:1013 arg1 domains 1024:1030 left-handed twisted receptor domains 995:1030 left-handed twisted receptor domains 995:1030 This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. 20145116 13 11 theme biochemical 1591:1601 arg1 analysis 1603:1610 This biochemical analysis 1586:1610 This biochemical analysis 1586:1610 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 10 12 theme D2/D3 1432:1436 arg1 linker 1438:1443 the D2/D3 linker 1428:1443 the D2/D3 linker 1428:1443 Additionally, the N-terminal helix of VEGF-C interacts with D2, and the groove separating the two VEGF-C monomers binds to the D2/D3 linker. 20145116 5 13 theme processed 641:649 arg1 VEGF-C 651:656 proteolytically processed VEGF-C 625:656 proteolytically processed VEGF-C 625:656 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 2 14 theme extracellular 251:263 arg1 domain 265:270 The extracellular domain 247:270 The extracellular domain of VEGF receptors 247:288 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 2 14 theme extracellular 251:263 arg1 receptors 280:288 VEGF receptors 275:288 VEGF receptors 275:288 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 13 15 dep together 1613:1620 arg1 with 1622:1625 with 1622:1625 with 1622:1625 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 14 16 theme significant 1740:1750 arg1 insights 1752:1759 significant insights 1740:1759 significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions 1740:1864 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 3 17 theme kinase 491:496 arg1 domain 498:503 the intracellular tyrosine kinase domain 464:503 the intracellular tyrosine kinase domain of the receptor 464:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 3 17 theme kinase 491:496 arg1 receptor 512:519 the receptor 508:519 the receptor 508:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 6 18 theme high-affinity-binding 848:868 arg1 site 870:873 the VEGFR-2 high-affinity-binding site 836:873 the VEGFR-2 high-affinity-binding site 836:873 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 1 19 theme vessel 154:159 arg1 formation 161:169 blood and lymph vessel formation 138:169 blood and lymph vessel formation 138:169 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 6 20 theme VEGFR-2 840:846 arg1 site 870:873 the VEGFR-2 high-affinity-binding site 836:873 the VEGFR-2 high-affinity-binding site 836:873 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 14 21 theme structural 1770:1779 arg1 features 1781:1788 the structural features 1766:1788 the structural features 1766:1788 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 14 22 theme interactions 1853:1864 arg1 specificity 1827:1837 specificity 1827:1837 specificity 1827:1837 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 14 22 theme interactions 1853:1864 arg1 affinity 1814:1821 affinity 1814:1821 affinity 1814:1821 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 7 23 theme left-handed 995:1005 arg1 domains 1024:1030 left-handed twisted receptor domains 995:1030 left-handed twisted receptor domains 995:1030 This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. 20145116 2 24 theme ligand 353:358 arg1 binding 360:366 ligand binding 353:366 ligand binding 353:366 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 0 25 theme Structural 0:9 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 0:83 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 20145116 13 26 theme VEGF-A 1701:1706 arg1 binding 1690:1696 the binding 1686:1696 the binding of VEGF-A and VEGF-C 1686:1717 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 1 27 theme Vascular 85:92 arg1 VEGFs 122:126 VEGFs 122:126 VEGFs 122:126 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 27 theme Vascular 85:92 arg1 factors 113:119 Vascular endothelial growth factors 85:119 Vascular endothelial growth factors (VEGFs) 85:127 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 14 28 dep affinity 1814:1821 arg1 the 1805:1807 the 1805:1807 the 1805:1807 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 13 29 theme VEGF-C 1712:1717 arg1 binding 1690:1696 the binding 1686:1696 the binding of VEGF-A and VEGF-C 1686:1717 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 0 30 theme growth 27:32 arg1 binding 41:47 growth factor binding 27:47 growth factor binding 27:47 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 20145116 8 31 theme N-terminal 1126:1135 arg1 helix 1143:1147 an N-terminal alpha helix 1123:1147 an N-terminal alpha helix 1123:1147 In the VEGFs, receptor specificity is determined by an N-terminal alpha helix and three peptide loops. 20145116 7 32 theme receptor 1015:1022 arg1 domains 1024:1030 left-handed twisted receptor domains 995:1030 left-handed twisted receptor domains 995:1030 This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. 20145116 2 33 theme VEGF 275:278 arg1 receptors 280:288 VEGF receptors 275:288 VEGF receptors 275:288 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 10 34 theme VEGF-C 1403:1408 arg1 monomers 1410:1417 the two VEGF-C monomers 1395:1417 the two VEGF-C monomers 1395:1417 Additionally, the N-terminal helix of VEGF-C interacts with D2, and the groove separating the two VEGF-C monomers binds to the D2/D3 linker. 20145116 2 35 theme receptors 280:288 arg1 domain 265:270 The extracellular domain 247:270 The extracellular domain of VEGF receptors 247:288 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 2 35 theme receptors 280:288 arg1 receptors 280:288 VEGF receptors 275:288 VEGF receptors 275:288 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 5 36 theme pathological 752:763 arg1 angiogenesis 765:776 physiological and pathological angiogenesis 734:776 physiological and pathological angiogenesis 734:776 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 1 37 theme growth 106:111 arg1 VEGFs 122:126 VEGFs 122:126 VEGFs 122:126 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 37 theme growth 106:111 arg1 factors 113:119 Vascular endothelial growth factors 85:119 Vascular endothelial growth factors (VEGFs) 85:127 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 10 38 theme N-terminal 1323:1332 arg1 helix 1334:1338 the N-terminal helix 1319:1338 the N-terminal helix of VEGF-C 1319:1348 Additionally, the N-terminal helix of VEGF-C interacts with D2, and the groove separating the two VEGF-C monomers binds to the D2/D3 linker. 20145116 0 39 theme binding 41:47 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 0:83 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 20145116 4 40 theme pathological 577:588 arg1 angiogenesis 590:601 pathological angiogenesis 577:601 pathological angiogenesis 577:601 VEGF-C stimulates lymphangiogenesis and contributes to pathological angiogenesis via VEGFR-3. 20145116 13 41 theme critical 1673:1680 arg1 residues 1664:1671 VEGFR-2 residues 1656:1671 VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C 1656:1717 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 1 42 theme receptor 199:206 arg1 VEGFR-1 226:232 VEGFR-1 226:232 VEGFR-1 226:232 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 42 theme receptor 199:206 arg1 kinases 217:223 three receptor tyrosine kinases 193:223 three receptor tyrosine kinases 193:223 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 42 theme receptor 199:206 arg1 -2 235:236 -2 235:236 -2 235:236 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 42 theme receptor 199:206 arg1 -3 243:244 -3 243:244 -3 243:244 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 2 43 theme receptor 377:384 arg1 dimerization 386:397 receptor dimerization 377:397 receptor dimerization 377:397 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 0 44 theme factor 34:39 arg1 binding 41:47 growth factor binding 27:47 growth factor binding 27:47 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 20145116 14 45 theme high 1809:1812 arg1 affinity 1814:1821 affinity 1814:1821 affinity 1814:1821 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 8 46 theme alpha 1137:1141 arg1 helix 1143:1147 an N-terminal alpha helix 1123:1147 an N-terminal alpha helix 1123:1147 In the VEGFs, receptor specificity is determined by an N-terminal alpha helix and three peptide loops. 20145116 6 47 theme VEGF-C 820:825 arg1 structure 807:815 the crystal structure 795:815 the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3 795:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 1 48 theme tyrosine 208:215 arg1 VEGFR-1 226:232 VEGFR-1 226:232 VEGFR-1 226:232 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 48 theme tyrosine 208:215 arg1 kinases 217:223 three receptor tyrosine kinases 193:223 three receptor tyrosine kinases 193:223 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 48 theme tyrosine 208:215 arg1 -2 235:236 -2 235:236 -2 235:236 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 48 theme tyrosine 208:215 arg1 -3 243:244 -3 243:244 -3 243:244 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 0 49 theme specificity 53:63 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 0:83 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 20145116 6 50 dep domains 918:924 arg1 D2 926:927 D2 926:927 D2 926:927 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 50 dep domains 918:924 arg1 domains 918:924 immunoglobulin homology domains D2 and D3 894:934 immunoglobulin homology domains D2 and D3 894:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 50 dep domains 918:924 arg1 D3 933:934 D3 933:934 D3 933:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 5 51 theme signals 713:719 arg1 VEGFR-2 674:680 VEGFR-2 674:680 VEGFR-2 674:680 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 5 51 theme signals 713:719 arg1 transducer 699:708 the predominant transducer 683:708 the predominant transducer of signals required for physiological and pathological angiogenesis 683:776 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 3 52 theme transmembrane 423:435 arg1 signaling 437:445 transmembrane signaling 423:445 transmembrane signaling 423:445 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 2 53 theme homology 323:330 arg1 domains 332:338 seven immunoglobulin homology domains 302:338 seven immunoglobulin homology domains 302:338 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 13 54 theme VEGFR-2 1656:1662 arg1 residues 1664:1671 VEGFR-2 residues 1656:1671 VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C 1656:1717 This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. 20145116 6 55 theme homology 909:916 arg1 D2 926:927 D2 926:927 D2 926:927 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 55 theme homology 909:916 arg1 domains 918:924 immunoglobulin homology domains D2 and D3 894:934 immunoglobulin homology domains D2 and D3 894:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 55 theme homology 909:916 arg1 D3 933:934 D3 933:934 D3 933:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 2 56 theme immunoglobulin 308:321 arg1 domains 332:338 seven immunoglobulin homology domains 302:338 seven immunoglobulin homology domains 302:338 The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. 20145116 6 57 theme immunoglobulin 894:907 arg1 D2 926:927 D2 926:927 D2 926:927 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 57 theme immunoglobulin 894:907 arg1 domains 918:924 immunoglobulin homology domains D2 and D3 894:934 immunoglobulin homology domains D2 and D3 894:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 57 theme immunoglobulin 894:907 arg1 D3 933:934 D3 933:934 D3 933:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 6 58 theme crystal 799:805 arg1 structure 807:815 the crystal structure 795:815 the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3 795:934 Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. 20145116 3 59 theme intracellular 468:480 arg1 domain 498:503 the intracellular tyrosine kinase domain 464:503 the intracellular tyrosine kinase domain of the receptor 464:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 3 59 theme intracellular 468:480 arg1 receptor 512:519 the receptor 508:519 the receptor 508:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 1 60 theme endothelial 94:104 arg1 VEGFs 122:126 VEGFs 122:126 VEGFs 122:126 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 60 theme endothelial 94:104 arg1 factors 113:119 Vascular endothelial growth factors 85:119 Vascular endothelial growth factors (VEGFs) 85:127 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 1 61 theme kinases 217:223 arg1 activation 179:188 activation 179:188 activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3 179:244 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 5 62 theme physiological 734:746 arg1 angiogenesis 765:776 physiological and pathological angiogenesis 734:776 physiological and pathological angiogenesis 734:776 However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. 20145116 7 63 theme 2-fold 1048:1053 arg1 axis 1055:1058 the 2-fold axis 1044:1058 the 2-fold axis of VEGF-C 1044:1068 This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. 20145116 3 64 theme tyrosine 482:489 arg1 domain 498:503 the intracellular tyrosine kinase domain 464:503 the intracellular tyrosine kinase domain of the receptor 464:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 3 64 theme tyrosine 482:489 arg1 receptor 512:519 the receptor 508:519 the receptor 508:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 10 65 theme VEGF-C 1343:1348 arg1 helix 1334:1338 the N-terminal helix 1319:1338 the N-terminal helix of VEGF-C 1319:1348 Additionally, the N-terminal helix of VEGF-C interacts with D2, and the groove separating the two VEGF-C monomers binds to the D2/D3 linker. 20145116 0 66 theme VEGF 68:71 arg1 receptor 73:80 VEGF receptor 2 68:82 VEGF receptor 2 68:82 Structural determinants of growth factor binding and specificity by VEGF receptor 2. 20145116 7 67 theme VEGF-C 1063:1068 arg1 axis 1055:1058 the 2-fold axis 1044:1058 the 2-fold axis of VEGF-C 1044:1068 This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. 20145116 14 68 theme VEGF/VEGFR 1842:1851 arg1 interactions 1853:1864 VEGF/VEGFR interactions 1842:1864 VEGF/VEGFR interactions 1842:1864 Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions. 20145116 3 69 theme receptor 512:519 arg1 domain 498:503 the intracellular tyrosine kinase domain 464:503 the intracellular tyrosine kinase domain of the receptor 464:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 3 69 theme receptor 512:519 arg1 receptor 512:519 the receptor 508:519 the receptor 508:519 Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. 20145116 7 70 theme symmetrical 962:972 arg1 complex 977:983 a symmetrical 22 complex 960:983 a symmetrical 22 complex 960:983 This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. 20145116 1 71 theme blood 138:142 arg1 formation 161:169 blood and lymph vessel formation 138:169 blood and lymph vessel formation 138:169 Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. 20145116 9 72 dep subdomains 1244:1253 arg1 subdomains 1244:1253 VEGFR-2 subdomains D2 and D3 1236:1263 VEGFR-2 subdomains D2 and D3 1236:1263 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 9 72 dep subdomains 1244:1253 arg1 D3 1262:1263 D3 1262:1263 D3 1262:1263 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 9 72 dep subdomains 1244:1253 arg1 D2 1255:1256 D2 1255:1256 D2 1255:1256 Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. 20145116 12 73 theme receptor 1564:1571 arg1 specificity 1573:1583 receptor specificity 1564:1583 receptor specificity 1564:1583 We therefore created VEGFR-1/VEGFR-2 chimeric proteins to further study receptor specificity. 23878260 9 0 theme VEGFR-specific 1631:1644 arg1 drugs 1646:1650 VEGFR-specific drugs 1631:1650 VEGFR-specific drugs 1631:1650 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 5 1 theme conserved 877:885 arg1 interface 902:910 a conserved ligand-binding interface 875:910 a conserved ligand-binding interface in D2 875:916 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 8 2 from interactions 1430:1441 arg1 D7 1453:1454 D7 1453:1454 D7 1453:1454 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 8 2 from interactions 1430:1441 arg1 D5 1446:1447 D5 1446:1447 D5 1446:1447 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 3 3 theme structural 651:660 arg1 rearrangements 662:675 structural rearrangements 651:675 structural rearrangements essential for receptor dimerization and activation 651:726 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 4 4 theme homodimer 840:848 arg1 structures 758:767 the crystal structures 746:767 the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer 746:848 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 5 5 theme ligand-binding 887:900 arg1 interface 902:910 a conserved ligand-binding interface 875:910 a conserved ligand-binding interface in D2 875:916 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 4 6 with complex 782:788 arg1 domains 803:809 VEGFR-3 domains D1-2 795:814 VEGFR-3 domains D1-2 795:814 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 4 7 theme VEGF-C 772:777 arg1 structures 758:767 the crystal structures 746:767 the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer 746:848 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 4 8 from structures 758:767 arg1 complex 782:788 complex 782:788 complex with VEGFR-3 domains D1-2 782:814 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 4 9 theme crystal 750:756 arg1 structures 758:767 the crystal structures 746:767 the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer 746:848 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 8 10 theme complex 1342:1348 arg1 consistent 1400:1409 consistent 1400:1409 consistent 1400:1409 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 8 10 theme complex 1342:1348 arg1 model 1309:1313 A structural model 1296:1313 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data 1296:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 7 11 theme thermodynamic 1180:1192 arg1 analysis 1194:1201 A thermodynamic analysis 1178:1201 A thermodynamic analysis of VEGFR-3 deletion mutants 1178:1229 A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. 23878260 8 12 theme structural 1298:1307 arg1 consistent 1400:1409 consistent 1400:1409 consistent 1400:1409 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 8 12 theme structural 1298:1307 arg1 model 1309:1313 A structural model 1296:1313 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data 1296:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 7 13 theme mutants 1223:1229 arg1 analysis 1194:1201 A thermodynamic analysis 1178:1201 A thermodynamic analysis of VEGFR-3 deletion mutants 1178:1229 A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. 23878260 3 14 dep domains 526:532 arg1 responsible 558:568 responsible 558:568 responsible 558:568 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 14 dep domains 526:532 arg1 involved 639:646 involved 639:646 are involved in structural rearrangements essential for receptor dimerization and activation 635:726 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 9 15 theme drugs 1646:1650 arg1 design 1621:1626 the design 1617:1626 the design of VEGFR-specific drugs 1617:1650 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 6 16 theme conserved 1031:1039 arg1 residues 1041:1048 the conserved residues 1027:1048 the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C 1027:1148 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 5 17 from interactions 995:1006 arg1 D5 1011:1012 D5 1011:1012 D5 1011:1012 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 8 18 attach derived 1350:1356 arg2 complex 1342:1348 the VEGF-C/VEGFR-3 D1-7 complex 1318:1348 the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data 1318:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 8 18 attach derived 1350:1356 arg1 data 1392:1395 small-angle X-ray scattering data 1363:1395 small-angle X-ray scattering data 1363:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 3 19 theme receptor 691:698 arg1 dimerization 700:711 receptor dimerization 691:711 receptor dimerization 691:711 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 6 20 theme compromised 1131:1141 arg1 VEGF-C 1143:1148 compromised VEGF-C 1131:1148 compromised VEGF-C 1131:1148 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 6 21 theme D5 1064:1065 arg1 interaction 1067:1077 the D5 interaction 1060:1077 the D5 interaction (Thr446 and Lys516) 1060:1097 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 4 22 theme D4-5 835:838 arg1 homodimer 840:848 the VEGFR-3 D4-5 homodimer 823:848 the VEGFR-3 D4-5 homodimer 823:848 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 8 23 theme D1-7 1337:1340 arg1 complex 1342:1348 the VEGF-C/VEGFR-3 D1-7 complex 1318:1348 the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data 1318:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 7 24 theme ligand 1280:1285 arg1 binding 1287:1293 ligand binding 1280:1293 ligand binding 1280:1293 A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. 23878260 0 25 theme Structural 0:9 arg1 insights 27:34 Structural and mechanistic insights 0:34 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 0:86 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 23878260 4 26 theme VEGFR-3 827:833 arg1 homodimer 840:848 the VEGFR-3 D4-5 homodimer 823:848 the VEGFR-3 D4-5 homodimer 823:848 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 5 27 from interface 902:910 arg1 D2 915:916 D2 915:916 D2 915:916 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 1 28 theme Vascular 88:95 arg1 VEGFs 125:129 VEGFs 125:129 VEGFs 125:129 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 28 theme Vascular 88:95 arg1 factors 116:122 Vascular endothelial growth factors 88:122 Vascular endothelial growth factors (VEGFs) 88:130 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 28 theme Vascular 88:95 arg1 receptors 142:150 their receptors 136:150 their receptors (VEGFRs) 136:159 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 28 theme Vascular 88:95 arg1 drivers 169:175 key drivers 165:175 key drivers of blood and lymph vessel formation in development, but also in several pathological processes 165:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 2 29 theme relevant 356:363 arg1 lymphangiogenesis 315:331 lymphangiogenesis 315:331 lymphangiogenesis 315:331 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 2 29 theme relevant 356:363 arg1 target 365:370 a clinically relevant target 343:370 a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis 343:458 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 2 30 theme tumor 426:430 arg1 angiogenesis 432:443 tumor angiogenesis 426:443 tumor angiogenesis 426:443 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 1 31 theme several 241:247 arg1 processes 262:270 several pathological processes 241:270 several pathological processes 241:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 2 32 theme VEGF-C 273:278 arg1 signaling 280:288 VEGF-C signaling 273:288 VEGF-C signaling through VEGFR-3 273:304 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 8 33 theme small-angle 1363:1373 arg1 scattering 1381:1390 small-angle X-ray scattering 1363:1390 small-angle X-ray scattering data 1363:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 5 34 theme VEGFR 945:949 arg1 dimerization 951:962 VEGFR dimerization 945:962 VEGFR dimerization 945:962 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 0 35 theme mechanistic 15:25 arg1 insights 27:34 Structural and mechanistic insights 0:34 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 0:86 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 23878260 1 36 theme pathological 249:260 arg1 processes 262:270 several pathological processes 241:270 several pathological processes 241:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 5 37 theme unique 924:929 arg1 mechanism 931:939 a unique mechanism 922:939 a unique mechanism for VEGFR dimerization and activation 922:977 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 6 38 theme VEGFR-3 1158:1164 arg1 activation 1166:1175 VEGFR-3 activation 1158:1175 VEGFR-3 activation 1158:1175 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 1 39 theme growth 109:114 arg1 VEGFs 125:129 VEGFs 125:129 VEGFs 125:129 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 39 theme growth 109:114 arg1 factors 116:122 Vascular endothelial growth factors 88:122 Vascular endothelial growth factors (VEGFs) 88:130 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 39 theme growth 109:114 arg1 receptors 142:150 their receptors 136:150 their receptors (VEGFRs) 136:159 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 39 theme growth 109:114 arg1 drivers 169:175 key drivers 165:175 key drivers of blood and lymph vessel formation in development, but also in several pathological processes 165:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 9 40 theme ligand-dependent 1492:1507 arg1 interactions 1519:1530 ligand-dependent homotypic interactions 1492:1530 ligand-dependent homotypic interactions in D5 and D7 1492:1543 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 3 41 theme extracellular 465:477 arg1 VEGFRs 489:494 VEGFRs 489:494 VEGFRs 489:494 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 41 theme extracellular 465:477 arg1 domain 479:484 The extracellular domain 461:484 The extracellular domain of VEGFRs 461:494 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 0 42 theme VEGF 41:44 arg1 binding 64:70 VEGF receptor 3 ligand binding 41:70 VEGF receptor 3 ligand binding 41:70 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 23878260 3 43 theme ligand 574:579 arg1 binding 581:587 ligand binding 574:587 ligand binding 574:587 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 1 44 theme key 165:167 arg1 factors 116:122 Vascular endothelial growth factors 88:122 Vascular endothelial growth factors (VEGFs) 88:130 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 44 theme key 165:167 arg1 receptors 142:150 their receptors 136:150 their receptors (VEGFRs) 136:159 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 44 theme key 165:167 arg1 drivers 169:175 key drivers 165:175 key drivers of blood and lymph vessel formation in development, but also in several pathological processes 165:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 45 theme blood 180:184 arg1 factors 116:122 Vascular endothelial growth factors 88:122 Vascular endothelial growth factors (VEGFs) 88:130 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 45 theme blood 180:184 arg1 receptors 142:150 their receptors 136:150 their receptors (VEGFRs) 136:159 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 45 theme blood 180:184 arg1 drivers 169:175 key drivers 165:175 key drivers of blood and lymph vessel formation in development, but also in several pathological processes 165:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 9 46 theme promising 1589:1597 arg1 possibilities 1599:1611 promising possibilities 1589:1611 promising possibilities for the design of VEGFR-specific drugs 1589:1650 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 4 47 theme VEGFR-3 795:801 arg1 domains 803:809 VEGFR-3 domains D1-2 795:814 VEGFR-3 domains D1-2 795:814 Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. 23878260 2 48 theme lymphatic 385:393 arg1 insufficiency 395:407 lymphatic insufficiency 385:407 lymphatic insufficiency 385:407 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 6 49 dep interaction 1067:1077 arg1 VEGF-C 1143:1148 compromised VEGF-C 1131:1148 compromised VEGF-C 1131:1148 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 6 49 dep interaction 1067:1077 arg1 Lys516 1091:1096 Lys516 1091:1096 Lys516 1091:1096 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 6 49 dep interaction 1067:1077 arg1 Thr446 1080:1085 Thr446 1080:1085 Thr446 1080:1085 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 7 50 theme deletion 1214:1221 arg1 mutants 1223:1229 VEGFR-3 deletion mutants 1206:1229 VEGFR-3 deletion mutants 1206:1229 A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. 23878260 6 51 theme D7 1107:1108 arg1 Arg737 1123:1128 Arg737 1123:1128 Arg737 1123:1128 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 6 51 theme D7 1107:1108 arg1 interaction 1110:1120 the D7 interaction 1103:1120 the D7 interaction (Arg737) 1103:1129 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 8 52 theme VEGF-C/VEGFR-3 1322:1335 arg1 complex 1342:1348 the VEGF-C/VEGFR-3 D1-7 complex 1318:1348 the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data 1318:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 3 53 theme VEGFRs 489:494 arg1 VEGFRs 489:494 VEGFRs 489:494 VEGFRs 489:494 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 53 theme VEGFRs 489:494 arg1 domain 479:484 The extracellular domain 461:484 The extracellular domain of VEGFRs 461:494 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 0 54 theme receptor 46:53 arg1 binding 64:70 VEGF receptor 3 ligand binding 41:70 VEGF receptor 3 ligand binding 41:70 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 23878260 8 55 theme scattering 1381:1390 arg1 data 1392:1395 small-angle X-ray scattering data 1363:1395 small-angle X-ray scattering data 1363:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 9 56 theme homotypic 1509:1517 arg1 interactions 1519:1530 ligand-dependent homotypic interactions 1492:1530 ligand-dependent homotypic interactions in D5 and D7 1492:1543 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 8 57 theme X-ray 1375:1379 arg1 scattering 1381:1390 small-angle X-ray scattering 1363:1390 small-angle X-ray scattering data 1363:1395 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 1 58 from drivers 169:175 arg1 processes 262:270 several pathological processes 241:270 several pathological processes 241:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 58 from drivers 169:175 arg1 development 216:226 development 216:226 development 216:226 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 5 59 from mechanism 931:939 arg1 D2 915:916 D2 915:916 D2 915:916 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 2 60 dep target 365:370 arg1 blocking 417:424 blocking 417:424 blocking tumor angiogenesis and metastasis 417:458 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 2 60 dep target 365:370 arg1 treating 376:383 treating 376:383 treating lymphatic insufficiency 376:407 VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. 23878260 1 61 dep blood 180:184 arg1 formation 203:211 vessel formation 196:211 vessel formation 196:211 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 9 62 theme VEGFR 1563:1567 arg1 activation 1569:1578 VEGFR activation 1563:1578 VEGFR activation 1563:1578 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 3 63 dep domains 535:541 arg1 D1-3 548:551 D1-3 548:551 D1-3 548:551 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 63 dep domains 535:541 arg1 1-3 543:545 1-3 543:545 1-3 543:545 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 1 64 theme endothelial 97:107 arg1 VEGFs 125:129 VEGFs 125:129 VEGFs 125:129 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 64 theme endothelial 97:107 arg1 factors 116:122 Vascular endothelial growth factors 88:122 Vascular endothelial growth factors (VEGFs) 88:130 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 64 theme endothelial 97:107 arg1 receptors 142:150 their receptors 136:150 their receptors (VEGFRs) 136:159 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 64 theme endothelial 97:107 arg1 drivers 169:175 key drivers 165:175 key drivers of blood and lymph vessel formation in development, but also in several pathological processes 165:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 65 theme lymph 190:194 arg1 factors 116:122 Vascular endothelial growth factors 88:122 Vascular endothelial growth factors (VEGFs) 88:130 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 65 theme lymph 190:194 arg1 receptors 142:150 their receptors 136:150 their receptors (VEGFRs) 136:159 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 1 65 theme lymph 190:194 arg1 drivers 169:175 key drivers 165:175 key drivers of blood and lymph vessel formation in development, but also in several pathological processes 165:270 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 8 66 theme homotypic 1420:1428 arg1 interactions 1430:1441 the homotypic interactions 1416:1441 the homotypic interactions in D5 and D7 1416:1454 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 23878260 3 67 theme membrane-proximal 598:614 arg1 domains 616:622 the membrane-proximal domains 4-7 594:626 the membrane-proximal domains 4-7 (D4-7) 594:633 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 67 theme membrane-proximal 598:614 arg1 D4-7 629:632 D4-7 629:632 D4-7 629:632 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 68 dep domains 616:622 arg1 4-7 624:626 4-7 624:626 4-7 624:626 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 1 69 theme vessel 196:201 arg1 formation 203:211 vessel formation 196:211 vessel formation 196:211 Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. 23878260 7 70 theme VEGFR-3 1206:1212 arg1 mutants 1223:1229 VEGFR-3 deletion mutants 1206:1229 VEGFR-3 deletion mutants 1206:1229 A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. 23878260 9 71 from interactions 1519:1530 arg1 D7 1542:1543 D7 1542:1543 D7 1542:1543 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 9 71 from interactions 1519:1530 arg1 D5 1535:1536 D5 1535:1536 D5 1535:1536 Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs. 23878260 5 72 theme homotypic 985:993 arg1 interactions 995:1006 homotypic interactions 985:1006 homotypic interactions in D5 985:1012 The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. 23878260 6 73 theme residues 1041:1048 arg1 Mutation 1015:1022 Mutation 1015:1022 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C 1015:1148 Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. 23878260 3 74 theme Ig 514:515 arg1 domains 526:532 seven Ig homology domains 508:532 seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation 508:726 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 0 75 theme ligand 57:62 arg1 binding 64:70 VEGF receptor 3 ligand binding 41:70 VEGF receptor 3 ligand binding 41:70 Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. 23878260 3 76 theme essential 677:685 arg1 rearrangements 662:675 structural rearrangements 651:675 structural rearrangements essential for receptor dimerization and activation 651:726 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 3 77 theme homology 517:524 arg1 domains 526:532 seven Ig homology domains 508:532 seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation 508:726 The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1-3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4-7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. 23878260 8 78 with consistent 1400:1409 arg1 interactions 1430:1441 the homotypic interactions 1416:1441 the homotypic interactions in D5 and D7 1416:1454 A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. 24338010 6 0 theme NPP4 1274:1277 arg1 specificity 1289:1299 NPP1 and NPP4 substrate specificity 1265:1299 specificity 1289:1299 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 4 1 theme NPP4 879:882 arg1 region 869:874 the corresponding region 851:874 the corresponding region of NPP4 851:882 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 6 2 theme rational 1325:1332 arg1 mechanism 1334:1342 a rational mechanism 1323:1342 a rational mechanism by which polymorphisms in NPP1 confer stroke resistance 1323:1398 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 1 3 theme extracellular 154:166 arg1 protein 177:183 a type I extracellular membrane protein 145:183 a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 145:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 3 theme extracellular 154:166 arg1 NPP4 137:140 NPP4 137:140 NPP4 137:140 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 4 theme Ap3A 267:270 arg1 hydrolysis 253:262 the hydrolysis 249:262 the hydrolysis of Ap3A 249:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 6 5 theme substrate 1279:1287 arg1 specificity 1289:1299 NPP1 and NPP4 substrate specificity 1265:1299 specificity 1289:1299 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 6 6 theme specificity 1289:1299 arg1 range 1305:1309 range 1305:1309 range 1305:1309 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 6 6 theme specificity 1289:1299 arg1 understanding 1248:1260 our understanding 1244:1260 our understanding of NPP1 and NPP4 substrate specificity 1244:1299 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 1 7 theme membrane 168:175 arg1 protein 177:183 a type I extracellular membrane protein 145:183 a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 145:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 7 theme membrane 168:175 arg1 NPP4 137:140 NPP4 137:140 NPP4 137:140 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 5 8 theme human 1089:1093 arg1 blood 1095:1099 human blood 1089:1099 human blood 1089:1099 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 2 9 theme high 544:547 arg1 structure 568:576 the high resolution crystal structure 540:576 the high resolution crystal structure of human NPP4 540:590 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 2 10 theme molecular 608:616 arg1 basis 618:622 the molecular basis 604:622 the molecular basis of its substrate specificity 604:651 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 4 11 theme ATP 959:961 arg1 hydrolysis 963:972 ATP hydrolysis 959:972 ATP hydrolysis 959:972 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 4 12 from claw 783:786 arg1 NPP1 791:794 NPP1 791:794 NPP1 791:794 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 4 13 theme lysine 776:781 arg1 claw 783:786 a tripartite lysine claw 763:786 a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis 763:972 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 1 14 theme chondrocytes 368:379 arg1 surface 357:363 the surface 353:363 the surface of chondrocytes 353:379 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 6 15 theme stroke 1382:1387 arg1 resistance 1389:1398 stroke resistance 1382:1398 stroke resistance 1382:1398 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 2 16 theme specificity 641:651 arg1 basis 618:622 the molecular basis 604:622 the molecular basis of its substrate specificity 604:651 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 5 17 theme platelet 1125:1132 arg1 aggregation 1134:1144 platelet aggregation 1125:1144 platelet aggregation 1125:1144 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 2 18 theme substrate 631:639 arg1 specificity 641:651 its substrate specificity 627:651 its substrate specificity 627:651 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 6 19 theme NPP1 1265:1268 arg1 specificity 1289:1299 NPP1 and NPP4 substrate specificity 1265:1299 specificity 1289:1299 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 1 20 theme brain 188:192 arg1 endothelium 203:213 brain vascular endothelium 188:213 brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 188:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 0 21 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis of purinergic signal metabolism by ectonucleotide 0:64 Molecular basis of purinergic signal metabolism by ectonucleotide pyrophosphatase/phosphodiesterases 4 and 1 and implications in stroke. 24338010 5 22 theme vascular 1194:1201 arg1 endothelium 1203:1213 vascular endothelium 1194:1213 vascular endothelium 1194:1213 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 4 23 theme corresponding 855:867 arg1 region 869:874 the corresponding region 851:874 the corresponding region of NPP4 851:882 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 1 24 theme vascular 194:201 arg1 endothelium 203:213 brain vascular endothelium 188:213 brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 188:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 0 25 theme purinergic 19:28 arg1 metabolism 37:46 purinergic signal metabolism 19:46 purinergic signal metabolism 19:46 Molecular basis of purinergic signal metabolism by ectonucleotide pyrophosphatase/phosphodiesterases 4 and 1 and implications in stroke. 24338010 6 26 from polymorphisms 1353:1365 arg1 NPP1 1370:1373 NPP1 1370:1373 NPP1 1370:1373 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 4 27 theme tripartite 765:774 arg1 claw 783:786 a tripartite lysine claw 763:786 a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis 763:972 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 2 28 theme purinergic 451:460 arg1 signals 462:468 purinergic signals 451:468 purinergic signals resulting in the physiologic activities of the two enzymes 451:527 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 1 29 theme type 291:294 arg1 protein 322:328 a type II extracellular membrane protein 289:328 a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization 289:416 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 29 theme type 291:294 arg1 NPP1 281:284 NPP1 281:284 NPP1 281:284 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 30 theme tissue 396:401 arg1 mineralization 403:416 tissue mineralization 396:416 tissue mineralization 396:416 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 0 31 theme metabolism 37:46 arg1 basis 10:14 Molecular basis 0:14 Molecular basis of purinergic signal metabolism by ectonucleotide 0:64 Molecular basis of purinergic signal metabolism by ectonucleotide pyrophosphatase/phosphodiesterases 4 and 1 and implications in stroke. 24338010 2 32 theme crystal 560:566 arg1 structure 568:576 the high resolution crystal structure 540:576 the high resolution crystal structure of human NPP4 540:590 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 5 33 from concentrations 1176:1189 arg1 endothelium 1203:1213 vascular endothelium 1194:1213 vascular endothelium 1194:1213 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 1 34 theme platelet 224:231 arg1 aggregation 233:243 platelet aggregation 224:243 platelet aggregation 224:243 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 0 35 theme signal 30:35 arg1 metabolism 37:46 purinergic signal metabolism 19:46 purinergic signal metabolism 19:46 Molecular basis of purinergic signal metabolism by ectonucleotide pyrophosphatase/phosphodiesterases 4 and 1 and implications in stroke. 24338010 1 36 theme extracellular 299:311 arg1 protein 322:328 a type II extracellular membrane protein 289:328 a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization 289:416 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 36 theme extracellular 299:311 arg1 NPP1 281:284 NPP1 281:284 NPP1 281:284 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 2 37 theme resolution 549:558 arg1 structure 568:576 the high resolution crystal structure 540:576 the high resolution crystal structure of human NPP4 540:590 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 0 38 from 4 101:101 arg1 stroke 129:134 stroke 129:134 stroke 129:134 Molecular basis of purinergic signal metabolism by ectonucleotide pyrophosphatase/phosphodiesterases 4 and 1 and implications in stroke. 24338010 2 39 theme signals 462:468 arg1 metabolism 437:446 the metabolism 433:446 the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes 433:527 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 2 40 theme physiologic 487:497 arg1 activities 499:508 the physiologic activities 483:508 the physiologic activities of the two enzymes 483:527 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 1 41 theme membrane 313:320 arg1 protein 322:328 a type II extracellular membrane protein 289:328 a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization 289:416 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 41 theme membrane 313:320 arg1 NPP1 281:284 NPP1 281:284 NPP1 281:284 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 42 dep protein 177:183 arg1 whereas 273:279 whereas 273:279 whereas 273:279 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 4 43 theme binding 919:925 arg1 orientation 927:937 this binding orientation 914:937 this binding orientation 914:937 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 5 44 theme physiologic 1050:1060 arg1 concentrations 1062:1075 physiologic concentrations 1050:1075 physiologic concentrations reported in human blood 1050:1099 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 4 45 theme ATP 838:840 arg1 phosphate 825:833 the terminal phosphate 812:833 the terminal phosphate of ATP 812:840 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 5 46 theme low 1162:1164 arg1 concentrations 1176:1189 low nanomolar concentrations 1162:1189 low nanomolar concentrations on vascular endothelium 1162:1213 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 1 47 attach present 342:348 arg2 NPP1 281:284 NPP1 281:284 NPP1 281:284 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 47 attach present 342:348 arg2 protein 322:328 a type II extracellular membrane protein 289:328 a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization 289:416 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 47 attach present 342:348 arg1 surface 357:363 the surface 353:363 the surface of chondrocytes 353:379 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 2 48 theme NPP4 587:590 arg1 structure 568:576 the high resolution crystal structure 540:576 the high resolution crystal structure of human NPP4 540:590 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 5 49 theme platelet 1026:1033 arg1 aggregation 1035:1045 platelet aggregation 1026:1045 platelet aggregation 1026:1045 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 24338010 4 50 theme structural 735:744 arg1 analysis 746:753 Comparative structural analysis 723:753 Comparative structural analysis 723:753 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 2 51 theme human 581:585 arg1 NPP4 587:590 human NPP4 581:590 human NPP4 581:590 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 4 52 theme Comparative 723:733 arg1 analysis 746:753 Comparative structural analysis 723:753 Comparative structural analysis 723:753 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 4 53 theme terminal 816:823 arg1 phosphate 825:833 the terminal phosphate 812:833 the terminal phosphate of ATP 812:840 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 1 54 theme type 147:150 arg1 protein 177:183 a type I extracellular membrane protein 145:183 a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 145:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 54 theme type 147:150 arg1 NPP4 137:140 NPP4 137:140 NPP4 137:140 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 2 55 theme enzymes 521:527 arg1 activities 499:508 the physiologic activities 483:508 the physiologic activities of the two enzymes 483:527 To understand the metabolism of purinergic signals resulting in the physiologic activities of the two enzymes, we report the high resolution crystal structure of human NPP4 and explore the molecular basis of its substrate specificity with NPP1. 24338010 1 56 theme present 342:348 arg1 protein 322:328 a type II extracellular membrane protein 289:328 a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization 289:416 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 56 theme present 342:348 arg1 NPP1 281:284 NPP1 281:284 NPP1 281:284 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 6 57 theme combined 1220:1227 arg1 studies 1229:1235 The combined studies 1216:1235 The combined studies 1216:1235 The combined studies expand our understanding of NPP1 and NPP4 substrate specificity and range and provide a rational mechanism by which polymorphisms in NPP1 confer stroke resistance. 24338010 1 58 from protein 177:183 arg1 endothelium 203:213 brain vascular endothelium 188:213 brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 188:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 59 theme I 152:152 arg1 protein 177:183 a type I extracellular membrane protein 145:183 a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A 145:270 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 1 59 theme I 152:152 arg1 NPP4 137:140 NPP4 137:140 NPP4 137:140 NPP4 is a type I extracellular membrane protein on brain vascular endothelium inducing platelet aggregation via the hydrolysis of Ap3A, whereas NPP1 is a type II extracellular membrane protein principally present on the surface of chondrocytes that regulates tissue mineralization. 24338010 4 60 contain contains 884:891 arg2 features 893:900 features 893:900 features that hinder this binding orientation 893:937 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 4 60 contain contains 884:891 arg1 region 869:874 the corresponding region 851:874 the corresponding region of NPP4 851:882 Comparative structural analysis reveals a tripartite lysine claw in NPP1 that stabilizes the terminal phosphate of ATP, whereas the corresponding region of NPP4 contains features that hinder this binding orientation, thereby inhibiting ATP hydrolysis. 24338010 5 61 theme nanomolar 1166:1174 arg1 concentrations 1176:1189 low nanomolar concentrations 1162:1189 low nanomolar concentrations on vascular endothelium 1162:1213 Furthermore, we show that NPP1 is unable to induce platelet aggregation at physiologic concentrations reported in human blood, but it could stimulate platelet aggregation if localized at low nanomolar concentrations on vascular endothelium. 18559974 6 0 theme dynamic 1010:1016 arg1 flap 1018:1021 the dynamic flap 1006:1021 the dynamic flap 1006:1021 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 18559974 0 1 theme TAFI 77:80 arg1 mechanism 54:62 the inactivation mechanism 37:62 the inactivation mechanism of activated TAFI 37:80 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 5 2 theme inhibitor 842:850 arg1 GEMSA 852:856 the inhibitor GEMSA 838:856 the inhibitor GEMSA 838:856 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 7 3 theme thrombin-cleavage 1250:1266 arg1 site 1268:1271 a cryptic thrombin-cleavage site 1240:1271 a cryptic thrombin-cleavage site present at Arg302 1240:1289 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 0 4 theme novel 85:89 arg1 mechanism 91:99 a novel mechanism 83:99 a novel mechanism for enzyme autoregulation 83:125 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 6 5 with interactions 1040:1051 arg1 peptide 1073:1079 the activation peptide 1058:1079 the activation peptide 1058:1079 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 18559974 8 6 theme control 1336:1342 arg1 mechanism 1316:1324 a novel mechanism 1308:1324 a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors 1308:1433 This represents a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors. 18559974 3 7 theme inhibitor 525:533 arg1 complex 543:549 a TAFI inhibitor (GEMSA) complex 518:549 a TAFI inhibitor (GEMSA) complex 518:549 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 7 8 theme flap 1134:1137 arg1 mobility 1139:1146 dynamic flap mobility 1126:1146 dynamic flap mobility 1126:1146 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 5 9 theme stabilizing 892:902 arg1 mutations 904:912 stabilizing mutations 892:912 stabilizing mutations 892:912 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 4 10 theme segment 717:723 arg1 dynamics 692:699 the dynamics 688:699 the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall 688:789 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 5 11 theme known 861:865 arg1 stabilizer 867:876 a known stabilizer 859:876 a known stabilizer of TAFIa 859:885 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 7 12 theme peptide 1108:1114 arg1 Release 1082:1088 Release 1082:1088 Release of the activation peptide 1082:1114 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 8 13 theme enzyme 1329:1334 arg1 control 1336:1342 enzyme control 1329:1342 enzyme control 1329:1342 This represents a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors. 18559974 6 14 theme TAFI 968:971 arg1 auto-regulation 973:987 TAFI auto-regulation 968:987 TAFI auto-regulation 968:987 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 18559974 4 15 theme crystal 620:626 arg1 structures 628:637 The crystal structures 616:637 The crystal structures 616:637 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 4 16 theme wall 786:789 arg1 wall 786:789 the active site wall 770:789 the active site wall 770:789 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 4 16 theme wall 786:789 arg1 residues 758:765 residues 758:765 residues of the active site wall 758:789 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 6 17 theme activation 1062:1071 arg1 peptide 1073:1079 the activation peptide 1058:1079 the activation peptide 1058:1079 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 18559974 3 18 theme TAFI 520:523 arg1 complex 543:549 a TAFI inhibitor (GEMSA) complex 518:549 a TAFI inhibitor (GEMSA) complex 518:549 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 7 19 from Arg302 1284:1289 arg1 present 1273:1279 present 1273:1279 present 1273:1279 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 5 20 theme flap 809:812 arg1 Dynamics 792:799 Dynamics 792:799 Dynamics of this flap 792:812 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 5 21 theme TAFIa 881:885 arg1 stabilizer 867:876 a known stabilizer 859:876 a known stabilizer of TAFIa 859:885 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 5 21 theme TAFIa 881:885 arg1 mutations 904:912 stabilizing mutations 892:912 stabilizing mutations 892:912 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 5 21 theme TAFIa 881:885 arg1 GEMSA 852:856 the inhibitor GEMSA 838:856 the inhibitor GEMSA 838:856 Dynamics of this flap are markedly reduced by the inhibitor GEMSA, a known stabilizer of TAFIa, and stabilizing mutations. 18559974 0 22 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of TAFI 0:25 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 8 23 theme inhibitors 1424:1433 arg1 absence 1404:1410 the absence 1400:1410 the absence of specific inhibitors 1400:1433 This represents a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors. 18559974 7 24 theme cryptic 1242:1248 arg1 site 1268:1271 a cryptic thrombin-cleavage site 1240:1271 a cryptic thrombin-cleavage site present at Arg302 1240:1289 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 0 25 theme TAFI 22:25 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of TAFI 0:25 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 4 26 theme active 774:779 arg1 wall 786:789 the active site wall 770:789 the active site wall 770:789 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 3 27 theme GEMSA 536:540 arg1 complex 543:549 a TAFI inhibitor (GEMSA) complex 518:549 a TAFI inhibitor (GEMSA) complex 518:549 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 2 28 theme clot 421:424 arg1 lysis 426:430 blood clot lysis 415:430 blood clot lysis 415:430 TAFIa is unique among carboxypeptidases in that it spontaneously inactivates with a short half-life, a property that is crucial for its role in controlling blood clot lysis. 18559974 0 29 theme enzyme 105:110 arg1 autoregulation 112:125 enzyme autoregulation 105:125 enzyme autoregulation 105:125 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 2 30 theme blood 415:419 arg1 lysis 426:430 blood clot lysis 415:430 blood clot lysis 415:430 TAFIa is unique among carboxypeptidases in that it spontaneously inactivates with a short half-life, a property that is crucial for its role in controlling blood clot lysis. 18559974 7 31 theme activation 1097:1106 arg1 peptide 1108:1114 the activation peptide 1093:1114 the activation peptide 1093:1114 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 3 32 theme TAFI 567:570 arg1 mutant 572:577 a quadruple TAFI mutant 555:577 a quadruple TAFI mutant (70-fold more stable active enzyme) 555:613 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 32 theme TAFI 567:570 arg1 enzyme 607:612 70-fold more stable active enzyme 580:612 70-fold more stable active enzyme 580:612 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 7 33 theme catalytic 1214:1222 arg1 site 1224:1227 the catalytic site 1210:1227 the catalytic site 1210:1227 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 4 34 theme TAFIa 649:653 arg1 stability 655:663 TAFIa stability 649:663 TAFIa stability 649:663 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 3 35 theme active 600:605 arg1 mutant 572:577 a quadruple TAFI mutant 555:577 a quadruple TAFI mutant (70-fold more stable active enzyme) 555:613 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 35 theme active 600:605 arg1 enzyme 607:612 70-fold more stable active enzyme 580:612 70-fold more stable active enzyme 580:612 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 36 theme intrinsic 448:456 arg1 instability 458:468 the intrinsic instability 444:468 the intrinsic instability of TAFIa 444:477 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 7 37 theme dynamic 1126:1132 arg1 mobility 1139:1146 dynamic flap mobility 1126:1146 dynamic flap mobility 1126:1146 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 4 38 theme site 781:784 arg1 wall 786:789 the active site wall 770:789 the active site wall 770:789 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 0 39 theme inactivation 41:52 arg1 mechanism 54:62 the inactivation mechanism 37:62 the inactivation mechanism of activated TAFI 37:80 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 3 40 theme quadruple 557:565 arg1 mutant 572:577 a quadruple TAFI mutant 555:577 a quadruple TAFI mutant (70-fold more stable active enzyme) 555:613 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 40 theme quadruple 557:565 arg1 enzyme 607:612 70-fold more stable active enzyme 580:612 70-fold more stable active enzyme 580:612 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 41 theme crystal 490:496 arg1 structures 498:507 crystal structures 490:507 crystal structures of TAFI 490:515 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 7 42 theme conformational 1174:1187 arg1 changes 1189:1195 conformational changes 1174:1195 conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302 1174:1289 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 1 43 theme fibrinolysis 149:160 arg1 inhibitor 162:170 Thrombin-activatable fibrinolysis inhibitor 128:170 Thrombin-activatable fibrinolysis inhibitor (TAFI) 128:177 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 1 43 theme fibrinolysis 149:160 arg1 pro-metallocarboxypeptidase 184:210 a pro-metallocarboxypeptidase 182:210 a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa) 182:256 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 1 43 theme fibrinolysis 149:160 arg1 TAFI 173:176 TAFI 173:176 TAFI 173:176 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 2 44 theme short 343:347 arg1 half-life 349:357 a short half-life 341:357 a short half-life 341:357 TAFIa is unique among carboxypeptidases in that it spontaneously inactivates with a short half-life, a property that is crucial for its role in controlling blood clot lysis. 18559974 2 44 theme short 343:347 arg1 property 362:369 a property 360:369 a property that is crucial for its role in controlling blood clot lysis 360:430 TAFIa is unique among carboxypeptidases in that it spontaneously inactivates with a short half-life, a property that is crucial for its role in controlling blood clot lysis. 18559974 2 44 theme short 343:347 arg1 crucial 379:385 crucial 379:385 crucial 379:385 TAFIa is unique among carboxypeptidases in that it spontaneously inactivates with a short half-life, a property that is crucial for its role in controlling blood clot lysis. 18559974 4 45 theme 55-residue 706:715 arg1 residues 726:733 residues 296-350 726:741 residues 296-350 726:741 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 4 45 theme 55-residue 706:715 arg1 segment 717:723 a 55-residue segment 704:723 a 55-residue segment (residues 296-350) that includes residues of the active site wall 704:789 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 3 46 theme stable 593:598 arg1 mutant 572:577 a quadruple TAFI mutant 555:577 a quadruple TAFI mutant (70-fold more stable active enzyme) 555:613 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 46 theme stable 593:598 arg1 enzyme 607:612 70-fold more stable active enzyme 580:612 70-fold more stable active enzyme 580:612 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 1 47 theme Thrombin-activatable 128:147 arg1 inhibitor 162:170 Thrombin-activatable fibrinolysis inhibitor 128:170 Thrombin-activatable fibrinolysis inhibitor (TAFI) 128:177 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 1 47 theme Thrombin-activatable 128:147 arg1 pro-metallocarboxypeptidase 184:210 a pro-metallocarboxypeptidase 182:210 a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa) 182:256 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 1 47 theme Thrombin-activatable 128:147 arg1 TAFI 173:176 TAFI 173:176 TAFI 173:176 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 1 48 dep activated 240:248 arg1 TAFIa 251:255 TAFIa 251:255 TAFIa 251:255 Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-metallocarboxypeptidase that can be proteolytically activated (TAFIa). 18559974 3 49 theme TAFIa 473:477 arg1 instability 458:468 the intrinsic instability 444:468 the intrinsic instability of TAFIa 444:477 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 0 50 theme activated 67:75 arg1 TAFI 77:80 activated TAFI 67:80 activated TAFI 67:80 Crystal structures of TAFI elucidate the inactivation mechanism of activated TAFI: a novel mechanism for enzyme autoregulation. 18559974 8 51 theme novel 1310:1314 arg1 mechanism 1316:1324 a novel mechanism 1308:1324 a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors 1308:1433 This represents a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors. 18559974 7 52 theme present 1273:1279 arg1 site 1268:1271 a cryptic thrombin-cleavage site 1240:1271 a cryptic thrombin-cleavage site present at Arg302 1240:1289 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 6 53 theme auto-regulation 973:987 arg1 model 959:963 a model 957:963 a model of TAFI auto-regulation 957:987 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 18559974 3 54 theme TAFI 512:515 arg1 mutant 572:577 a quadruple TAFI mutant 555:577 a quadruple TAFI mutant (70-fold more stable active enzyme) 555:613 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 54 theme TAFI 512:515 arg1 complex 543:549 a TAFI inhibitor (GEMSA) complex 518:549 a TAFI inhibitor (GEMSA) complex 518:549 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 54 theme TAFI 512:515 arg1 enzyme 607:612 70-fold more stable active enzyme 580:612 70-fold more stable active enzyme 580:612 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 3 54 theme TAFI 512:515 arg1 structures 498:507 crystal structures 490:507 crystal structures of TAFI 490:515 We studied the intrinsic instability of TAFIa by solving crystal structures of TAFI, a TAFI inhibitor (GEMSA) complex and a quadruple TAFI mutant (70-fold more stable active enzyme). 18559974 6 55 theme zymogen 993:999 arg1 TAFI 1001:1004 zymogen TAFI 993:1004 zymogen TAFI 993:1004 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 18559974 8 56 from activity 1378:1385 arg1 plasma 1390:1395 plasma 1390:1395 plasma 1390:1395 This represents a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors. 18559974 8 57 theme specific 1415:1422 arg1 inhibitors 1424:1433 specific inhibitors 1415:1433 specific inhibitors 1415:1433 This represents a novel mechanism of enzyme control that enables TAFI to regulate its activity in plasma in the absence of specific inhibitors. 18559974 7 58 attach present 1273:1279 arg2 site 1268:1271 a cryptic thrombin-cleavage site 1240:1271 a cryptic thrombin-cleavage site present at Arg302 1240:1289 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 7 58 attach present 1273:1279 arg1 Arg302 1284:1289 Arg302 1284:1289 Arg302 1284:1289 Release of the activation peptide increases dynamic flap mobility and in time this leads to conformational changes that disrupt the catalytic site and expose a cryptic thrombin-cleavage site present at Arg302. 18559974 4 59 dep residues 726:733 arg1 296-350 735:741 296-350 735:741 296-350 735:741 The crystal structures show that TAFIa stability is directly related to the dynamics of a 55-residue segment (residues 296-350) that includes residues of the active site wall. 18559974 6 60 theme structural 936:945 arg1 basis 947:951 the structural basis 932:951 the structural basis for a model of TAFI auto-regulation 932:987 Our data provide the structural basis for a model of TAFI auto-regulation: in zymogen TAFI the dynamic flap is stabilized by interactions with the activation peptide. 3840370 7 0 theme protein 1090:1096 arg1 Examination 1071:1081 Examination 1071:1081 Examination of the protein 1071:1096 Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. 3840370 7 0 theme protein 1090:1096 arg1 results 1116:1122 cDNA sequence results 1102:1122 cDNA sequence results 1102:1122 Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. 3840370 10 1 dep long 1418:1421 arg1 kilobases 1408:1416 kilobases 1408:1416 kilobases 1408:1416 2.5 kilobases long. 3840370 2 2 theme entire 333:338 arg1 sequence 351:358 The entire amino acid sequence 329:358 The entire amino acid sequence of the C4b-binding protein 329:385 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 1 3 theme cDNA 246:249 arg1 portions 234:241 portions 234:241 portions of cDNA coding for human C4b-binding protein 234:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 12 4 dep tryptophan 1645:1654 arg1 acids 1555:1559 60 amino acids long 1546:1564 60 amino acids long 1546:1564 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 13 5 from regions 1700:1706 arg1 protein 1723:1729 C4b-binding protein 1711:1729 C4b-binding protein 1711:1729 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 13 6 theme non-complement 1935:1948 arg1 2-glycoprotein 1955:1968 the non-complement beta 2-glycoprotein I 1931:1970 the non-complement beta 2-glycoprotein I 1931:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 6 7 theme 000 1028:1030 arg1 chains 1003:1008 chains 1003:1008 chains of apparent Mr 70 000 that contains 549 amino acid residues 1003:1068 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 11 8 theme C4b-binding 1458:1468 arg1 protein 1470:1476 C4b-binding protein 1458:1476 C4b-binding protein 1458:1476 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 13 9 theme C4b-binding 1711:1721 arg1 protein 1723:1729 C4b-binding protein 1711:1729 C4b-binding protein 1711:1729 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 6 10 theme Mr 1022:1023 arg1 000 1028:1030 apparent Mr 70 000 1013:1030 apparent Mr 70 000 1013:1030 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 1 11 attach isolated 293:300 arg1 library 320:326 a liver cDNA library 307:326 a liver cDNA library 307:326 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 1 11 attach isolated 293:300 arg2 clones 216:221 plasmid clones 208:221 plasmid clones containing portions of cDNA coding for human C4b-binding protein 208:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 1 12 theme synthetic 170:178 arg1 probes 200:205 probes 200:205 probes 200:205 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 1 12 theme synthetic 170:178 arg1 oligonucleotides 180:195 synthetic oligonucleotides 170:195 synthetic oligonucleotides 170:195 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 9 13 theme C4b-binding 1373:1383 arg1 protein 1385:1391 C4b-binding protein 1373:1391 C4b-binding protein 1373:1391 Northern-blot analysis indicated that the mRNA for C4b-binding protein is approx. 3840370 2 14 theme [Chung 501:506 arg1 Reid 518:521 Reid 518:521 Reid 518:521 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 2 14 theme [Chung 501:506 arg1 level 495:499 the protein level [Chung 483:506 the protein level [Chung 483:506 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 0 15 theme pathway 121:127 arg1 protein 74:80 C4b-binding protein 62:80 C4b-binding protein 62:80 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 0 15 theme pathway 121:127 arg1 protein 96:102 a regulatory protein 83:102 a regulatory protein of the classical pathway of the human complement system 83:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 13 16 theme internal-homology 1896:1912 arg1 regions 1914:1920 the internal-homology regions 1892:1920 the internal-homology regions found in the non-complement beta 2-glycoprotein I 1892:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 1 17 contain containing 223:232 arg2 portions 234:241 portions 234:241 portions of cDNA coding for human C4b-binding protein 234:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 1 17 contain containing 223:232 arg1 clones 216:221 plasmid clones 208:221 plasmid clones containing portions of cDNA coding for human C4b-binding protein 208:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 5 18 theme sequence 750:757 arg1 nucleotides 730:740 1717 nucleotides 725:740 1717 nucleotides of cDNA sequence 725:757 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 6 19 located found 971:975 arg1 plasma 980:985 plasma 980:985 plasma 980:985 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 6 19 located found 971:975 arg2 form 942:945 the secreted form 929:945 the secreted form 929:945 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 13 20 theme factor 1871:1876 arg1 B 1878:1878 the complement enzyme factor B 1849:1878 the complement enzyme factor B 1849:1878 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 4 21 theme N-terminal 618:627 arg1 residues 632:639 the N-terminal 62 residues 614:639 the N-terminal 62 residues 614:639 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. 3840370 0 22 theme complement 142:151 arg1 system 153:158 the human complement system 132:158 the human complement system 132:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 5 23 from codon 771:775 arg1 region 890:895 the 3' non-translated region 868:895 the 3' non-translated region 868:895 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 6 24 theme sequence 902:909 arg1 studies 911:917 The sequence studies 898:917 The sequence studies 898:917 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 2 25 theme sequence 465:472 arg1 study 474:478 a previous sequence study 454:478 a previous sequence study 454:478 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 6 26 theme amino 1050:1054 arg1 residues 1061:1068 549 amino acid residues 1046:1068 549 amino acid residues 1046:1068 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 4 27 theme amino 583:587 arg1 sequence 594:601 the amino acid sequence 579:601 the amino acid sequence 579:601 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. 3840370 1 28 theme cDNA 315:318 arg1 library 320:326 a liver cDNA library 307:326 a liver cDNA library 307:326 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 5 29 theme 32nd 785:788 arg1 acid 796:799 the 32nd amino acid 781:799 the 32nd amino acid in the sequence of C4b-binding protein 781:838 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 4 30 theme sequence 594:601 arg1 sequence 594:601 the amino acid sequence 579:601 the amino acid sequence 579:601 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. 3840370 4 30 theme sequence 594:601 arg1 % 574:574 55% 572:574 55% of the amino acid sequence 572:601 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. 3840370 5 31 theme plasmid 660:666 arg1 clones 668:673 The plasmid clones 656:673 The plasmid clones isolated 656:682 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 13 32 theme complement 1853:1862 arg1 B 1878:1878 the complement enzyme factor B 1849:1878 the complement enzyme factor B 1849:1878 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 2 33 theme protein 487:493 arg1 Reid 518:521 Reid 518:521 Reid 518:521 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 2 33 theme protein 487:493 arg1 level 495:499 the protein level [Chung 483:506 the protein level [Chung 483:506 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 2 34 theme protein 379:385 arg1 sequence 351:358 The entire amino acid sequence 329:358 The entire amino acid sequence of the C4b-binding protein 329:385 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 13 35 theme Ba 1836:1837 arg1 region 1839:1844 the Ba region 1832:1844 the Ba region of the complement enzyme factor B 1832:1878 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 12 36 theme several 1660:1666 arg1 residues 1684:1691 several other conserved residues 1660:1691 several other conserved residues 1660:1691 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 6 37 theme C4b-binding 950:960 arg1 protein 962:968 C4b-binding protein 950:968 C4b-binding protein 950:968 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 0 38 theme Molecular 0:8 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning 0:16 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 1 39 theme human 262:266 arg1 protein 280:286 human C4b-binding protein 262:286 human C4b-binding protein 262:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 12 40 theme conserved 1674:1682 arg1 residues 1684:1691 several other conserved residues 1660:1691 several other conserved residues 1660:1691 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 5 41 theme protein 832:838 arg1 sequence 808:815 the sequence 804:815 the sequence of C4b-binding protein 804:838 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 13 42 with homologous 1735:1744 arg1 regions 1779:1785 the three internal-homology regions 1751:1785 the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I 1751:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 13 42 with homologous 1735:1744 arg1 present 1817:1823 present 1817:1823 present 1817:1823 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 11 43 theme internal 1504:1511 arg1 regions 1524:1530 eight internal homologous regions 1498:1530 eight internal homologous regions 1498:1530 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 11 43 theme internal 1504:1511 arg1 each 1533:1536 each 1533:1536 each 1533:1536 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 13 44 gly 2-glycoprotein 1955:1968 arg1 2-glycoprotein 1955:1968 the non-complement beta 2-glycoprotein I 1931:1970 the non-complement beta 2-glycoprotein I 1931:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 2 45 theme cDNA 434:437 arg1 study 414:418 this study 409:418 this study of the cloned cDNA 409:437 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 0 46 theme cDNA 46:49 arg1 characterization 22:37 characterization 22:37 characterization 22:37 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 0 46 theme cDNA 46:49 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning 0:16 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 1 47 theme plasmid 208:214 arg1 clones 216:221 plasmid clones 208:221 plasmid clones containing portions of cDNA coding for human C4b-binding protein 208:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 11 48 theme N-terminal 1428:1437 arg1 acids 1449:1453 The N-terminal 491 amino acids 1424:1453 The N-terminal 491 amino acids of C4b-binding protein 1424:1476 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 7 49 theme sequence 1107:1114 arg1 results 1116:1122 cDNA sequence results 1102:1122 cDNA sequence results 1102:1122 Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. 3840370 6 50 contain contains 1037:1044 arg1 chains 1003:1008 chains 1003:1008 chains of apparent Mr 70 000 that contains 549 amino acid residues 1003:1068 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 6 50 contain contains 1037:1044 arg2 residues 1061:1068 549 amino acid residues 1046:1068 549 amino acid residues 1046:1068 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 2 51 theme amino 340:344 arg1 sequence 351:358 The entire amino acid sequence 329:358 The entire amino acid sequence of the C4b-binding protein 329:385 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 12 52 theme long 1561:1564 arg1 acids 1555:1559 60 amino acids long 1546:1564 60 amino acids long 1546:1564 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 11 53 theme amino 1443:1447 arg1 acids 1449:1453 The N-terminal 491 amino acids 1424:1453 The N-terminal 491 amino acids of C4b-binding protein 1424:1476 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 5 54 from nucleotide 854:863 arg1 region 890:895 the 3' non-translated region 868:895 the 3' non-translated region 868:895 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 9 55 theme Northern-blot 1322:1334 arg1 analysis 1336:1343 Northern-blot analysis 1322:1343 Northern-blot analysis 1322:1343 Northern-blot analysis indicated that the mRNA for C4b-binding protein is approx. 3840370 13 56 theme internal-homology 1761:1777 arg1 regions 1779:1785 the three internal-homology regions 1751:1785 the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I 1751:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 13 56 theme internal-homology 1761:1777 arg1 present 1817:1823 present 1817:1823 present 1817:1823 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 11 57 theme protein 1470:1476 arg1 acids 1449:1453 The N-terminal 491 amino acids 1424:1453 The N-terminal 491 amino acids of C4b-binding protein 1424:1476 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 12 58 theme amino 1549:1553 arg1 acids 1555:1559 60 amino acids long 1546:1564 60 amino acids long 1546:1564 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 6 59 theme apparent 1013:1020 arg1 000 1028:1030 apparent Mr 70 000 1013:1030 apparent Mr 70 000 1013:1030 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 0 60 theme classical 111:119 arg1 pathway 121:127 the classical pathway 107:127 the classical pathway of the human complement system 107:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 5 61 theme nucleotides 730:740 arg1 determination 708:720 the unambiguous determination 692:720 the unambiguous determination of 1717 nucleotides of cDNA sequence 692:757 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 13 62 located present 1817:1823 arg2 present 1817:1823 present 1817:1823 present 1817:1823 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 13 62 located present 1817:1823 arg2 regions 1779:1785 the three internal-homology regions 1751:1785 the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I 1751:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 13 62 located present 1817:1823 arg1 region 1839:1844 the Ba region 1832:1844 the Ba region of the complement enzyme factor B 1832:1878 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 5 63 theme cDNA 745:748 arg1 sequence 750:757 cDNA sequence 745:757 cDNA sequence 745:757 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 0 64 theme human 136:140 arg1 system 153:158 the human complement system 132:158 the human complement system 132:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 0 65 theme system 153:158 arg1 pathway 121:127 the classical pathway 107:127 the classical pathway of the human complement system 107:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 13 66 theme B 1878:1878 arg1 region 1839:1844 the Ba region 1832:1844 the Ba region of the complement enzyme factor B 1832:1878 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 6 67 theme acid 1056:1059 arg1 residues 1061:1068 549 amino acid residues 1046:1068 549 amino acid residues 1046:1068 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 0 68 theme C4b-binding 62:72 arg1 protein 74:80 C4b-binding protein 62:80 C4b-binding protein 62:80 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 0 68 theme C4b-binding 62:72 arg1 protein 96:102 a regulatory protein 83:102 a regulatory protein of the classical pathway of the human complement system 83:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 2 69 theme previous 456:463 arg1 study 474:478 a previous sequence study 454:478 a previous sequence study 454:478 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 13 70 theme beta 1950:1953 arg1 2-glycoprotein 1955:1968 the non-complement beta 2-glycoprotein I 1931:1970 the non-complement beta 2-glycoprotein I 1931:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 4 71 from 22 544:545 arg1 which 561:565 which 561:565 which 561:565 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. 3840370 5 72 theme amino 790:794 arg1 acid 796:799 the 32nd amino acid 781:799 the 32nd amino acid in the sequence of C4b-binding protein 781:838 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 4 73 theme acid 589:592 arg1 sequence 594:601 the amino acid sequence 579:601 the amino acid sequence 579:601 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. 3840370 0 74 theme regulatory 85:94 arg1 protein 74:80 C4b-binding protein 62:80 C4b-binding protein 62:80 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 0 74 theme regulatory 85:94 arg1 protein 96:102 a regulatory protein 83:102 a regulatory protein of the classical pathway of the human complement system 83:158 Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system. 3840370 13 75 theme enzyme 1864:1869 arg1 B 1878:1878 the complement enzyme factor B 1849:1878 the complement enzyme factor B 1849:1878 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 5 76 from acid 796:799 arg1 sequence 808:815 the sequence 804:815 the sequence of C4b-binding protein 804:838 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 5 77 theme 164th 848:852 arg1 nucleotide 854:863 the 164th nucleotide 844:863 the 164th nucleotide in the 3' non-translated region 844:895 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 12 78 theme other 1668:1672 arg1 residues 1684:1691 several other conserved residues 1660:1691 several other conserved residues 1660:1691 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 6 79 theme protein 962:968 arg1 form 942:945 the secreted form 929:945 the secreted form 929:945 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 2 80 theme C4b-binding 367:377 arg1 protein 379:385 the C4b-binding protein 363:385 the C4b-binding protein 363:385 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 5 81 theme unambiguous 696:706 arg1 determination 708:720 the unambiguous determination 692:720 the unambiguous determination of 1717 nucleotides of cDNA sequence 692:757 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 13 82 located found 1922:1926 arg2 regions 1914:1920 the internal-homology regions 1892:1920 the internal-homology regions found in the non-complement beta 2-glycoprotein I 1892:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 13 82 located found 1922:1926 arg1 2-glycoprotein 1955:1968 the non-complement beta 2-glycoprotein I 1931:1970 the non-complement beta 2-glycoprotein I 1931:1970 These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I. 3840370 5 83 theme C4b-binding 820:830 arg1 protein 832:838 C4b-binding protein 820:838 C4b-binding protein 820:838 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 7 84 theme polymorphic 1157:1167 arg1 sites 1169:1173 at least two polymorphic sites 1144:1173 at least two polymorphic sites in the molecule 1144:1189 Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. 3840370 1 85 theme C4b-binding 268:278 arg1 protein 280:286 human C4b-binding protein 262:286 human C4b-binding protein 262:286 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 6 86 theme secreted 933:940 arg1 form 942:945 the secreted form 929:945 the secreted form 929:945 The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. 3840370 2 87 theme cloned 427:432 arg1 cDNA 434:437 the cloned cDNA 423:437 the cloned cDNA 423:437 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 3840370 1 88 theme liver 309:313 arg1 library 320:326 a liver cDNA library 307:326 a liver cDNA library 307:326 By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. 3840370 12 89 theme half-cystine 1627:1638 arg1 region 1612:1617 each region 1607:1617 each region of four half-cystine 1607:1638 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 12 90 from presence 1595:1602 arg1 region 1612:1617 each region 1607:1617 each region of four half-cystine 1607:1638 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. 3840370 11 91 theme homologous 1513:1522 arg1 regions 1524:1530 eight internal homologous regions 1498:1530 eight internal homologous regions 1498:1530 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 11 91 theme homologous 1513:1522 arg1 each 1533:1536 each 1533:1536 each 1533:1536 The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 3840370 5 92 theme non-translated 875:888 arg1 region 890:895 the 3' non-translated region 868:895 the 3' non-translated region 868:895 The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3' non-translated region. 3840370 7 93 from sites 1169:1173 arg1 molecule 1182:1189 the molecule 1178:1189 the molecule 1178:1189 Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. 3840370 7 94 theme cDNA 1102:1105 arg1 results 1116:1122 cDNA sequence results 1102:1122 cDNA sequence results 1102:1122 Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. 3840370 2 95 theme acid 346:349 arg1 sequence 351:358 The entire amino acid sequence 329:358 The entire amino acid sequence of the C4b-binding protein 329:385 The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. 6203908 3 0 theme human 137:141 arg1 2-macroglobulin 149:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 8 1 theme reactive 786:793 arg1 site 795:798 an activatable reactive site 771:798 an activatable reactive site which can mediate covalent binding of nucleophiles 771:849 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 13 2 from stretches 1349:1357 arg1 2-macroglobulin 1383:1397 alpha 2-macroglobulin 1377:1397 alpha 2-macroglobulin 1377:1397 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 8 3 theme beta-SH 673:679 arg1 thiol 701:705 thiol 701:705 thiol esterified to the gamma-carbonyl group of Glx-952 701:755 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 8 3 theme beta-SH 673:679 arg1 Cys-949 690:696 Cys-949 690:696 Cys-949 690:696 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 8 3 theme beta-SH 673:679 arg1 group 681:685 The beta-SH group 669:685 The beta-SH group of Cys-949 669:696 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 10 4 from sites 950:954 arg1 area 1007:1010 the activation cleavage area 983:1010 the activation cleavage area (the "bait" region) 983:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 4 from sites 950:954 arg1 region 1024:1029 the "bait" region 1013:1029 the "bait" region 1013:1029 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 14 5 theme systems 1620:1626 arg1 context 1583:1589 the context 1579:1589 the context of proteolytically regulated systems with particular reference to the complement components C3 and C4 1579:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 14 6 theme complement 1661:1670 arg1 components 1672:1681 the complement components C3 and C4 1657:1691 the complement components C3 and C4 1657:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 14 6 theme complement 1661:1670 arg1 C4 1690:1691 C4 1690:1691 C4 1690:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 14 6 theme complement 1661:1670 arg1 C3 1683:1684 C3 1683:1684 C3 1683:1684 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 8 7 theme nucleophiles 838:849 arg1 binding 827:833 covalent binding 818:833 covalent binding of nucleophiles 818:849 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 14 8 theme regulated 1610:1618 arg1 systems 1620:1626 proteolytically regulated systems 1594:1626 proteolytically regulated systems with particular reference to the complement components C3 and C4 1594:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 13 9 theme complement 1430:1439 arg1 components 1441:1450 complement components C3 and C4 1430:1460 complement components C3 and C4 1430:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 13 9 theme complement 1430:1439 arg1 C4 1459:1460 C4 1459:1460 C4 1459:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 13 9 theme complement 1430:1439 arg1 C3 1452:1453 C3 1452:1453 C3 1452:1453 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 4 10 theme acid 228:231 arg1 residues 233:240 1451 amino acid residues 217:240 1451 amino acid residues 217:240 The identical subunits contain 1451 amino acid residues. 6203908 13 11 theme stretches 1349:1357 arg1 comparison 1335:1344 A comparison 1333:1344 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 1333:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 6 12 dep placed 409:414 arg1 Cys1056-Cys1104 551:565 Cys1056-Cys1104 551:565 Cys1056-Cys1104 551:565 Eleven intrachain disulfide bridges have been placed (Cys25-Cys63, Cys228-Cys276, Cys246-Cys264, Cys255-Cys408, Cys572-Cys748, Cys619-Cys666, Cys798-Cys826, Cys824-Cys860, Cys898-Cys1298, Cys1056-Cys1104, and Cys1329-Cys1444). 6203908 6 12 dep placed 409:414 arg1 Cys1329-Cys1444 572:586 Cys1329-Cys1444 572:586 Cys1329-Cys1444 572:586 Eleven intrachain disulfide bridges have been placed (Cys25-Cys63, Cys228-Cys276, Cys246-Cys264, Cys255-Cys408, Cys572-Cys748, Cys619-Cys666, Cys798-Cys826, Cys824-Cys860, Cys898-Cys1298, Cys1056-Cys1104, and Cys1329-Cys1444). 6203908 7 13 theme interchain 616:625 arg1 bridge 627:632 an interchain bridge 613:632 an interchain bridge with Cys-447 from another subunit 613:666 Cys-447 probably forms an interchain bridge with Cys-447 from another subunit. 6203908 10 14 theme sequence 1051:1058 arg1 -Arg681-Val-Gly-Phe-Tyr-Glu- 1061:1088 the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu- 1047:1088 the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu- 1047:1088 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 14 15 theme particular 1633:1642 arg1 reference 1644:1652 particular reference 1633:1652 particular reference to the complement components C3 and C4 1633:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 12 16 theme alpha 1289:1293 arg1 subunit 1311:1317 the alpha 2-macroglobulin subunit 1285:1317 the alpha 2-macroglobulin subunit 1285:1317 The presence of possible internal homologies within the alpha 2-macroglobulin subunit is discussed. 6203908 10 17 from cleavage 971:978 arg1 area 1007:1010 the activation cleavage area 983:1010 the activation cleavage area (the "bait" region) 983:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 17 from cleavage 971:978 arg1 region 1024:1029 the "bait" region 1013:1029 the "bait" region 1013:1029 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 18 from located 1036:1042 arg1 -Arg681-Val-Gly-Phe-Tyr-Glu- 1061:1088 the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu- 1047:1088 the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu- 1047:1088 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 14 19 dep components 1672:1681 arg1 components 1672:1681 the complement components C3 and C4 1657:1691 the complement components C3 and C4 1657:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 14 19 dep components 1672:1681 arg1 C4 1690:1691 C4 1690:1691 C4 1690:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 14 19 dep components 1672:1681 arg1 C3 1683:1684 C3 1683:1684 C3 1683:1684 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 8 20 theme Cys-949 690:696 arg1 thiol 701:705 thiol 701:705 thiol esterified to the gamma-carbonyl group of Glx-952 701:755 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 8 20 theme Cys-949 690:696 arg1 Cys-949 690:696 Cys-949 690:696 Cys-949 690:696 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 8 20 theme Cys-949 690:696 arg1 group 681:685 The beta-SH group 669:685 The beta-SH group of Cys-949 669:696 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 13 21 theme sequence 1412:1419 arg1 data 1421:1424 partial sequence data 1404:1424 partial sequence data for complement components C3 and C4 1404:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 4 22 theme identical 190:198 arg1 subunits 200:207 The identical subunits 186:207 The identical subunits 186:207 The identical subunits contain 1451 amino acid residues. 6203908 5 23 theme asparagine 300:309 arg1 residues 311:318 asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401 300:360 asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401 300:360 Glucosamine-based oligosaccharide groups are attached to asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401. 6203908 4 24 contain contain 209:215 arg2 residues 233:240 1451 amino acid residues 217:240 1451 amino acid residues 217:240 The identical subunits contain 1451 amino acid residues. 6203908 4 24 contain contain 209:215 arg1 subunits 200:207 The identical subunits 186:207 The identical subunits 186:207 The identical subunits contain 1451 amino acid residues. 6203908 10 25 theme proteolytic 959:969 arg1 cleavage 971:978 proteolytic cleavage 959:978 proteolytic cleavage in the activation cleavage area (the "bait" region) 959:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 3 26 gly glycoprotein 124:135 arg1 glycoprotein 124:135 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 0 27 theme Primary 0:6 arg1 structure 8:16 Primary structure 0:16 Primary structure of human alpha 2-macroglobulin 0:47 Primary structure of human alpha 2-macroglobulin. 6203908 6 28 theme disulfide 381:389 arg1 bridges 391:397 Eleven intrachain disulfide bridges 363:397 Eleven intrachain disulfide bridges 363:397 Eleven intrachain disulfide bridges have been placed (Cys25-Cys63, Cys228-Cys276, Cys246-Cys264, Cys255-Cys408, Cys572-Cys748, Cys619-Cys666, Cys798-Cys826, Cys824-Cys860, Cys898-Cys1298, Cys1056-Cys1104, and Cys1329-Cys1444). 6203908 3 29 theme primary 81:87 arg1 structure 89:97 The primary structure 77:97 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin 77:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 10 30 theme " 1022:1022 arg1 area 1007:1010 the activation cleavage area 983:1010 the activation cleavage area (the "bait" region) 983:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 30 theme " 1022:1022 arg1 region 1024:1029 the "bait" region 1013:1029 the "bait" region 1013:1029 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 0 31 theme human 21:25 arg1 2-macroglobulin 33:47 human alpha 2-macroglobulin 21:47 human alpha 2-macroglobulin 21:47 Primary structure of human alpha 2-macroglobulin. 6203908 2 32 theme complete 57:64 arg1 structure 66:74 The complete structure 53:74 The complete structure 53:74 The complete structure. 6203908 9 33 theme putative 854:861 arg1 site 894:897 A putative transglutaminase cross-linking site 852:897 A putative transglutaminase cross-linking site 852:897 A putative transglutaminase cross-linking site is constituted by Gln-670 and Gln-671. 6203908 10 34 theme activation 987:996 arg1 area 1007:1010 the activation cleavage area 983:1010 the activation cleavage area (the "bait" region) 983:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 34 theme activation 987:996 arg1 region 1024:1029 the "bait" region 1013:1029 the "bait" region 1013:1029 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 11 35 theme molecular 1095:1103 arg1 160,837 1163:1169 160,837 1163:1169 160,837 1163:1169 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 11 35 theme molecular 1095:1103 arg1 weight 1105:1110 The molecular weight 1091:1110 The molecular weight of the unmodified alpha 2-macroglobulin subunit 1091:1158 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 11 36 theme unmodified 1119:1128 arg1 subunit 1152:1158 the unmodified alpha 2-macroglobulin subunit 1115:1158 the unmodified alpha 2-macroglobulin subunit 1115:1158 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 9 37 theme transglutaminase 863:878 arg1 site 894:897 A putative transglutaminase cross-linking site 852:897 A putative transglutaminase cross-linking site 852:897 A putative transglutaminase cross-linking site is constituted by Gln-670 and Gln-671. 6203908 10 38 theme primary 942:948 arg1 sites 950:954 The primary sites 938:954 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) 938:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 38 theme primary 942:948 arg1 cleavage 971:978 proteolytic cleavage 959:978 proteolytic cleavage in the activation cleavage area (the "bait" region) 959:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 38 theme primary 942:948 arg1 located 1036:1042 located 1036:1042 located 1036:1042 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 39 from area 1007:1010 arg1 sites 950:954 The primary sites 938:954 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) 938:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 39 from area 1007:1010 arg1 cleavage 971:978 proteolytic cleavage 959:978 proteolytic cleavage in the activation cleavage area (the "bait" region) 959:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 39 from area 1007:1010 arg1 located 1036:1042 located 1036:1042 located 1036:1042 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 0 40 theme 2-macroglobulin 33:47 arg1 structure 8:16 Primary structure 0:16 Primary structure of human alpha 2-macroglobulin 0:47 Primary structure of human alpha 2-macroglobulin. 6203908 12 41 theme possible 1249:1256 arg1 homologies 1267:1276 possible internal homologies 1249:1276 possible internal homologies 1249:1276 The presence of possible internal homologies within the alpha 2-macroglobulin subunit is discussed. 6203908 9 42 theme cross-linking 880:892 arg1 site 894:897 A putative transglutaminase cross-linking site 852:897 A putative transglutaminase cross-linking site 852:897 A putative transglutaminase cross-linking site is constituted by Gln-670 and Gln-671. 6203908 5 43 theme Glucosamine-based 243:259 arg1 groups 277:282 Glucosamine-based oligosaccharide groups 243:282 Glucosamine-based oligosaccharide groups 243:282 Glucosamine-based oligosaccharide groups are attached to asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401. 6203908 6 44 theme intrachain 370:379 arg1 bridges 391:397 Eleven intrachain disulfide bridges 363:397 Eleven intrachain disulfide bridges 363:397 Eleven intrachain disulfide bridges have been placed (Cys25-Cys63, Cys228-Cys276, Cys246-Cys264, Cys255-Cys408, Cys572-Cys748, Cys619-Cys666, Cys798-Cys826, Cys824-Cys860, Cys898-Cys1298, Cys1056-Cys1104, and Cys1329-Cys1444). 6203908 8 45 theme covalent 818:825 arg1 binding 827:833 covalent binding 818:833 covalent binding of nucleophiles 818:849 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 0 46 theme alpha 27:31 arg1 2-macroglobulin 33:47 human alpha 2-macroglobulin 21:47 human alpha 2-macroglobulin 21:47 Primary structure of human alpha 2-macroglobulin. 6203908 8 47 theme activatable 774:784 arg1 site 795:798 an activatable reactive site 771:798 an activatable reactive site which can mediate covalent binding of nucleophiles 771:849 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 5 48 theme oligosaccharide 261:275 arg1 groups 277:282 Glucosamine-based oligosaccharide groups 243:282 Glucosamine-based oligosaccharide groups 243:282 Glucosamine-based oligosaccharide groups are attached to asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401. 6203908 10 49 from -Arg681-Val-Gly-Phe-Tyr-Glu- 1061:1088 arg1 sites 950:954 The primary sites 938:954 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) 938:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 49 from -Arg681-Val-Gly-Phe-Tyr-Glu- 1061:1088 arg1 cleavage 971:978 proteolytic cleavage 959:978 proteolytic cleavage in the activation cleavage area (the "bait" region) 959:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 49 from -Arg681-Val-Gly-Phe-Tyr-Glu- 1061:1088 arg1 located 1036:1042 located 1036:1042 located 1036:1042 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 3 50 theme tetrameric 106:115 arg1 2-macroglobulin 149:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 12 51 theme 2-macroglobulin 1295:1309 arg1 subunit 1311:1317 the alpha 2-macroglobulin subunit 1285:1317 the alpha 2-macroglobulin subunit 1285:1317 The presence of possible internal homologies within the alpha 2-macroglobulin subunit is discussed. 6203908 8 52 theme gamma-carbonyl 725:738 arg1 group 740:744 the gamma-carbonyl group 721:744 the gamma-carbonyl group of Glx-952 721:755 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 8 52 theme gamma-carbonyl 725:738 arg1 Glx-952 749:755 Glx-952 749:755 Glx-952 749:755 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 11 53 theme carbohydrate 1212:1223 arg1 groups 1225:1230 the carbohydrate groups 1208:1230 the carbohydrate groups 1208:1230 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 14 54 with systems 1620:1626 arg1 reference 1644:1652 particular reference 1633:1652 particular reference to the complement components C3 and C4 1633:1691 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 3 55 theme plasma 117:122 arg1 2-macroglobulin 149:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 12 56 theme internal 1258:1265 arg1 homologies 1267:1276 possible internal homologies 1249:1276 possible internal homologies 1249:1276 The presence of possible internal homologies within the alpha 2-macroglobulin subunit is discussed. 6203908 8 57 theme Glx-952 749:755 arg1 group 740:744 the gamma-carbonyl group 721:744 the gamma-carbonyl group of Glx-952 721:755 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 8 57 theme Glx-952 749:755 arg1 Glx-952 749:755 Glx-952 749:755 Glx-952 749:755 The beta-SH group of Cys-949 is thiol esterified to the gamma-carbonyl group of Glx-952, thus forming an activatable reactive site which can mediate covalent binding of nucleophiles. 6203908 13 58 from 2-macroglobulin 1383:1397 arg1 stretches 1349:1357 stretches 1349:1357 stretches of sequences from alpha 2-macroglobulin 1349:1397 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 13 58 from 2-macroglobulin 1383:1397 arg1 sequences 1362:1370 sequences 1362:1370 sequences from alpha 2-macroglobulin 1362:1397 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 10 59 theme cleavage 971:978 arg1 sites 950:954 The primary sites 938:954 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) 938:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 59 theme cleavage 971:978 arg1 cleavage 971:978 proteolytic cleavage 959:978 proteolytic cleavage in the activation cleavage area (the "bait" region) 959:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 59 theme cleavage 971:978 arg1 located 1036:1042 located 1036:1042 located 1036:1042 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 13 60 with comparison 1335:1344 arg1 data 1421:1424 partial sequence data 1404:1424 partial sequence data for complement components C3 and C4 1404:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 10 61 theme bait 1018:1021 arg1 area 1007:1010 the activation cleavage area 983:1010 the activation cleavage area (the "bait" region) 983:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 61 theme bait 1018:1021 arg1 region 1024:1029 the "bait" region 1013:1029 the "bait" region 1013:1029 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 62 located located 1036:1042 arg2 located 1036:1042 located 1036:1042 located 1036:1042 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 62 located located 1036:1042 arg2 sites 950:954 The primary sites 938:954 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) 938:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 62 located located 1036:1042 arg1 -Arg681-Val-Gly-Phe-Tyr-Glu- 1061:1088 the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu- 1047:1088 the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu- 1047:1088 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 62 located located 1036:1042 arg2 cleavage 971:978 proteolytic cleavage 959:978 proteolytic cleavage in the activation cleavage area (the "bait" region) 959:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 11 63 theme subunit 1152:1158 arg1 160,837 1163:1169 160,837 1163:1169 160,837 1163:1169 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 11 63 theme subunit 1152:1158 arg1 weight 1105:1110 The molecular weight 1091:1110 The molecular weight of the unmodified alpha 2-macroglobulin subunit 1091:1158 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 11 64 theme alpha 1130:1134 arg1 subunit 1152:1158 the unmodified alpha 2-macroglobulin subunit 1115:1158 the unmodified alpha 2-macroglobulin subunit 1115:1158 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 14 65 theme 2-macroglobulin 1542:1556 arg1 properties 1522:1531 The properties 1518:1531 The properties of alpha 2-macroglobulin 1518:1556 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 13 66 theme sequences 1362:1370 arg1 stretches 1349:1357 stretches 1349:1357 stretches of sequences from alpha 2-macroglobulin 1349:1397 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 3 67 theme alpha 143:147 arg1 2-macroglobulin 149:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 12 68 theme homologies 1267:1276 arg1 presence 1237:1244 The presence 1233:1244 The presence of possible internal homologies within the alpha 2-macroglobulin subunit 1233:1317 The presence of possible internal homologies within the alpha 2-macroglobulin subunit is discussed. 6203908 14 69 theme alpha 1536:1540 arg1 2-macroglobulin 1542:1556 alpha 2-macroglobulin 1536:1556 alpha 2-macroglobulin 1536:1556 The properties of alpha 2-macroglobulin are discussed within the context of proteolytically regulated systems with particular reference to the complement components C3 and C4. 6203908 7 70 from subunit 660:666 arg1 Cys-447 639:645 Cys-447 639:645 Cys-447 from another subunit 639:666 Cys-447 probably forms an interchain bridge with Cys-447 from another subunit. 6203908 11 71 theme 2-macroglobulin 1136:1150 arg1 subunit 1152:1158 the unmodified alpha 2-macroglobulin subunit 1115:1158 the unmodified alpha 2-macroglobulin subunit 1115:1158 The molecular weight of the unmodified alpha 2-macroglobulin subunit is 160,837 and approximately 179,000, including the carbohydrate groups. 6203908 3 72 theme 2-macroglobulin 149:163 arg1 structure 89:97 The primary structure 77:97 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin 77:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 5 73 attach attached 288:295 arg2 groups 277:282 Glucosamine-based oligosaccharide groups 243:282 Glucosamine-based oligosaccharide groups 243:282 Glucosamine-based oligosaccharide groups are attached to asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401. 6203908 5 73 attach attached 288:295 arg1 residues 311:318 asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401 300:360 asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401 300:360 Glucosamine-based oligosaccharide groups are attached to asparagine residues 32, 47, 224, 373, 387, 846, 968, and 1401. 6203908 7 74 with bridge 627:632 arg1 Cys-447 639:645 Cys-447 639:645 Cys-447 from another subunit 639:666 Cys-447 probably forms an interchain bridge with Cys-447 from another subunit. 6203908 13 75 theme alpha 1377:1381 arg1 2-macroglobulin 1383:1397 alpha 2-macroglobulin 1377:1397 alpha 2-macroglobulin 1377:1397 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 13 76 dep components 1441:1450 arg1 components 1441:1450 complement components C3 and C4 1430:1460 complement components C3 and C4 1430:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 13 76 dep components 1441:1450 arg1 C4 1459:1460 C4 1459:1460 C4 1459:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 13 76 dep components 1441:1450 arg1 C3 1452:1453 C3 1452:1453 C3 1452:1453 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 6203908 10 77 theme cleavage 998:1005 arg1 area 1007:1010 the activation cleavage area 983:1010 the activation cleavage area (the "bait" region) 983:1030 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 10 77 theme cleavage 998:1005 arg1 region 1024:1029 the "bait" region 1013:1029 the "bait" region 1013:1029 The primary sites of proteolytic cleavage in the activation cleavage area (the "bait" region) are located in the sequence: -Arg681-Val-Gly-Phe-Tyr-Glu-. 6203908 3 78 theme glycoprotein 124:135 arg1 2-macroglobulin 149:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 the tetrameric plasma glycoprotein human alpha 2-macroglobulin 102:163 The primary structure of the tetrameric plasma glycoprotein human alpha 2-macroglobulin has been determined. 6203908 4 79 theme amino 222:226 arg1 residues 233:240 1451 amino acid residues 217:240 1451 amino acid residues 217:240 The identical subunits contain 1451 amino acid residues. 6203908 13 80 theme partial 1404:1410 arg1 data 1421:1424 partial sequence data 1404:1424 partial sequence data for complement components C3 and C4 1404:1460 A comparison of stretches of sequences from alpha 2-macroglobulin with partial sequence data for complement components C3 and C4 indicates that these proteins are evolutionary related. 25043189 2 0 theme factor 456:461 arg1 receptors 489:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 5 1 theme hit 1114:1116 arg1 blockade 1137:1144 a "double hit" cytokine receptor blockade 1104:1144 a "double hit" cytokine receptor blockade 1104:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 2 2 theme anti-leukemia 313:325 arg1 antibody 327:334 the anti-leukemia antibody CSL362 309:341 the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations 309:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 4 3 theme open-like 825:833 arg1 mutants 842:848 "open-like" IL3Rα mutants 824:848 "open-like" IL3Rα mutants 824:848 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 3 4 theme alternative 749:759 arg1 mechanisms 761:770 alternative mechanisms 749:770 alternative mechanisms for blocking IL-3 signaling 749:798 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 2 5 theme colony-stimulating 437:454 arg1 GM-CSF 464:469 GM-CSF 464:469 GM-CSF 464:469 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 5 theme colony-stimulating 437:454 arg1 factor 456:461 the granulocyte-macrophage colony-stimulating factor 410:461 the granulocyte-macrophage colony-stimulating factor (GM-CSF) 410:470 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 5 6 theme potent 1069:1074 arg1 antibodies 1076:1085 potent antibodies 1069:1085 potent antibodies that can achieve a "double hit" cytokine receptor blockade 1069:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 2 7 theme classical 527:535 arg1 conformations 546:558 classical "closed" conformations 527:558 classical "closed" conformations 527:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 1 8 theme innate 143:148 arg1 immunity 163:170 innate and adaptive immunity 143:170 innate and adaptive immunity 143:170 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 5 9 theme receptor 1128:1135 arg1 blockade 1137:1144 a "double hit" cytokine receptor blockade 1104:1144 a "double hit" cytokine receptor blockade 1104:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 3 10 theme CSL362 622:627 arg1 epitope 611:617 the NTD epitope 603:617 the NTD epitope of CSL362 603:627 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 2 11 theme receptor 268:275 arg1 chain 279:283 the IL-3 receptor α chain 259:283 the IL-3 receptor α chain (IL3Rα) 259:291 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 11 theme receptor 268:275 arg1 IL3Rα 286:290 IL3Rα 286:290 IL3Rα 286:290 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 12 theme IL-5 473:476 arg1 receptors 489:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 13 attach present 399:405 arg1 receptors 489:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 13 attach present 399:405 arg2 domain 371:376 the N-terminal domain 356:376 the N-terminal domain (NTD) 356:382 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 13 attach present 399:405 arg2 domain 387:392 a domain 385:392 a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 385:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 14 theme IL-3 263:266 arg1 chain 279:283 the IL-3 receptor α chain 259:283 the IL-3 receptor α chain (IL3Rα) 259:291 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 14 theme IL-3 263:266 arg1 IL3Rα 286:290 IL3Rα 286:290 IL3Rα 286:290 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 15 from present 399:405 arg1 receptors 489:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 16 theme N-terminal 360:369 arg1 domain 387:392 a domain 385:392 a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 385:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 16 theme N-terminal 360:369 arg1 NTD 379:381 NTD 379:381 NTD 379:381 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 16 theme N-terminal 360:369 arg1 domain 371:376 the N-terminal domain 356:376 the N-terminal domain (NTD) 356:382 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 17 theme granulocyte-macrophage 414:435 arg1 GM-CSF 464:469 GM-CSF 464:469 GM-CSF 464:469 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 17 theme granulocyte-macrophage 414:435 arg1 factor 456:461 the granulocyte-macrophage colony-stimulating factor 410:461 the granulocyte-macrophage colony-stimulating factor (GM-CSF) 410:470 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 4 18 theme IL-3 944:947 arg1 complex 968:974 a higher-order IL-3 receptor-signaling complex 929:974 a higher-order IL-3 receptor-signaling complex 929:974 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 1 19 theme adaptive 154:161 arg1 immunity 163:170 innate and adaptive immunity 143:170 innate and adaptive immunity 143:170 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 4 20 theme receptor-signaling 949:966 arg1 complex 968:974 a higher-order IL-3 receptor-signaling complex 929:974 a higher-order IL-3 receptor-signaling complex 929:974 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 5 21 theme " 1117:1117 arg1 blockade 1137:1144 a "double hit" cytokine receptor blockade 1104:1144 a "double hit" cytokine receptor blockade 1104:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 2 22 theme " 544:544 arg1 conformations 546:558 classical "closed" conformations 527:558 classical "closed" conformations 527:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 3 23 theme IL-3 657:660 arg1 site 670:673 the IL-3 binding site 653:673 the IL-3 binding site 653:673 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 0 24 theme Dual 0:3 arg1 mechanism 5:13 Dual mechanism 0:13 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody 0:75 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody. 25043189 5 25 theme cytokine 1119:1126 arg1 blockade 1137:1144 a "double hit" cytokine receptor blockade 1104:1144 a "double hit" cytokine receptor blockade 1104:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 2 26 theme closed 538:543 arg1 conformations 546:558 classical "closed" conformations 527:558 classical "closed" conformations 527:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 0 27 theme interleukin-3 18:30 arg1 blockade 41:48 interleukin-3 receptor blockade 18:48 interleukin-3 receptor blockade 18:48 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody. 25043189 5 28 theme double 1107:1112 arg1 blockade 1137:1144 a "double hit" cytokine receptor blockade 1104:1144 a "double hit" cytokine receptor blockade 1104:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 5 29 theme receptors 1017:1025 arg1 forms 999:1003 open forms 994:1003 open forms of cytokine receptors 994:1025 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 5 30 theme forms 999:1003 arg1 discovery 981:989 The discovery 977:989 The discovery of open forms of cytokine receptors 977:1025 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 2 31 theme chain 279:283 arg1 structure 246:254 the crystal structure 234:254 the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations 234:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 3 32 theme extensive 570:578 arg1 analyses 591:598 extensive mutational analyses 570:598 extensive mutational analyses of the NTD epitope of CSL362 570:627 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 4 33 theme complex 968:974 arg1 assembly 917:924 the assembly 913:924 the assembly of a higher-order IL-3 receptor-signaling complex 913:974 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 3 34 theme binding 662:668 arg1 site 670:673 the IL-3 binding site 653:673 the IL-3 binding site 653:673 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 3 35 theme mutational 580:589 arg1 analyses 591:598 extensive mutational analyses 570:598 extensive mutational analyses of the NTD epitope of CSL362 570:627 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 2 36 theme α 277:277 arg1 chain 279:283 the IL-3 receptor α chain 259:283 the IL-3 receptor α chain (IL3Rα) 259:291 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 36 theme α 277:277 arg1 IL3Rα 286:290 IL3Rα 286:290 IL3Rα 286:290 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 37 from structure 246:254 arg1 complex 296:302 complex 296:302 complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations 296:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 0 38 theme blockade 41:48 arg1 mechanism 5:13 Dual mechanism 0:13 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody 0:75 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody. 25043189 3 39 dep minor 634:638 arg1 overlap 640:646 overlap 640:646 overlap 640:646 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 2 40 theme IL-13 483:487 arg1 receptors 489:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 410:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 1 41 theme several 191:197 arg1 immunopathologies 199:215 several immunopathologies 191:215 several immunopathologies 191:215 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 0 42 theme receptor 32:39 arg1 blockade 41:48 interleukin-3 receptor blockade 18:48 interleukin-3 receptor blockade 18:48 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody. 25043189 3 43 theme IL-3 697:700 arg1 binding 702:708 IL-3 binding 697:708 IL-3 binding to the closed conformation 697:735 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 5 44 theme cytokine 1008:1015 arg1 receptors 1017:1025 cytokine receptors 1008:1025 cytokine receptors 1008:1025 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 4 45 theme higher-order 931:942 arg1 complex 968:974 a higher-order IL-3 receptor-signaling complex 929:974 a higher-order IL-3 receptor-signaling complex 929:974 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 2 46 theme open 517:520 arg1 " 521:521 unique "open" 509:521 unique "open" 509:521 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 47 with complex 296:302 arg1 antibody 327:334 the anti-leukemia antibody CSL362 309:341 the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations 309:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 4 48 theme IL3Rα 836:840 arg1 mutants 842:848 "open-like" IL3Rα mutants 824:848 "open-like" IL3Rα mutants 824:848 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 2 49 theme present 399:405 arg1 domain 387:392 a domain 385:392 a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors 385:497 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 2 49 theme present 399:405 arg1 domain 371:376 the N-terminal domain 356:376 the N-terminal domain (NTD) 356:382 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 3 50 theme NTD 607:609 arg1 epitope 611:617 the NTD epitope 603:617 the NTD epitope of CSL362 603:627 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 1 51 theme activated 105:113 arg1 Interleukin-3 78:90 Interleukin-3 78:90 Interleukin-3 (IL-3) 78:97 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 1 51 theme activated 105:113 arg1 product 122:128 an activated T cell product 102:128 an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies 102:215 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 3 52 theme IL-3 785:788 arg1 signaling 790:798 blocking IL-3 signaling 776:798 blocking IL-3 signaling 776:798 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 2 53 theme unique 509:514 arg1 " 521:521 unique "open" 509:521 unique "open" 509:521 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 3 54 theme epitope 611:617 arg1 analyses 591:598 extensive mutational analyses 570:598 extensive mutational analyses of the NTD epitope of CSL362 570:627 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 1 55 theme T 115:115 arg1 Interleukin-3 78:90 Interleukin-3 78:90 Interleukin-3 (IL-3) 78:97 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 1 55 theme T 115:115 arg1 product 122:128 an activated T cell product 102:128 an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies 102:215 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 3 56 theme closed 717:722 arg1 conformation 724:735 the closed conformation 713:735 the closed conformation 713:735 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 4 57 theme " 834:834 arg1 mutants 842:848 "open-like" IL3Rα mutants 824:848 "open-like" IL3Rα mutants 824:848 Significantly, whereas "open-like" IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. 25043189 2 58 theme crystal 238:244 arg1 structure 246:254 the crystal structure 234:254 the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations 234:558 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 25043189 1 59 theme cell 117:120 arg1 Interleukin-3 78:90 Interleukin-3 78:90 Interleukin-3 (IL-3) 78:97 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 1 59 theme cell 117:120 arg1 product 122:128 an activated T cell product 102:128 an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies 102:215 Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. 25043189 5 60 theme open 994:997 arg1 forms 999:1003 open forms 994:1003 open forms of cytokine receptors 994:1025 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 5 61 theme antibodies 1076:1085 arg1 development 1054:1064 development 1054:1064 development of potent antibodies that can achieve a "double hit" cytokine receptor blockade 1054:1144 The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a "double hit" cytokine receptor blockade. 25043189 3 62 theme blocking 776:783 arg1 signaling 790:798 blocking IL-3 signaling 776:798 blocking IL-3 signaling 776:798 Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. 25043189 0 63 theme anti-cancer 56:66 arg1 antibody 68:75 an anti-cancer antibody 53:75 an anti-cancer antibody 53:75 Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody. 25043189 2 64 from receptors 489:497 arg1 present 399:405 present 399:405 present 399:405 Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique "open" and classical "closed" conformations. 12144777 4 0 theme barrel 504:509 arg1 subunits 511:518 the (alpha/beta)(8) barrel subunits 484:518 the (alpha/beta)(8) barrel subunits of the homodimeric molecule 484:546 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 6 1 theme catalytic 1062:1070 arg1 domain 1072:1077 the catalytic domain 1058:1077 the catalytic domain of the bacterial urease 1058:1101 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 1 theme catalytic 1062:1070 arg1 urease 1096:1101 the bacterial urease 1082:1101 the bacterial urease 1082:1101 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 5 2 theme dipeptide 885:893 arg1 substrates 895:904 dipeptide substrates 885:904 dipeptide substrates 885:904 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 5 3 theme used 761:764 arg1 inhibitor 777:785 the therapeutically used cilastatin inhibitor 741:785 the therapeutically used cilastatin inhibitor 741:785 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 4 4 theme microvillar 677:687 arg1 membrane 689:696 the microvillar membrane 673:696 the microvillar membrane of a kidney tubule 673:715 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 2 5 theme unliganded 325:334 arg1 structures 263:272 The crystal structures 251:272 The crystal structures of the saccharide-trimmed enzyme 251:305 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 2 5 theme unliganded 325:334 arg1 forms 359:363 unliganded and inhibitor-liganded forms 325:363 unliganded and inhibitor-liganded forms 325:363 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 4 6 theme Glu125 598:603 arg1 side-chain 605:614 the Glu125 side-chain 594:614 the Glu125 side-chain located at the bottom of the barrel 594:650 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 5 7 theme dipeptidyl 720:729 arg1 moiety 731:736 A dipeptidyl moiety 718:736 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor 718:785 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 6 8 theme urease 1096:1101 arg1 similarities 998:1009 unexpected similarities 987:1009 unexpected similarities to those of the murine adenosine deaminase 987:1052 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 8 theme urease 1096:1101 arg1 domain 1072:1077 the catalytic domain 1058:1077 the catalytic domain of the bacterial urease 1058:1101 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 8 theme urease 1096:1101 arg1 urease 1096:1101 the bacterial urease 1082:1101 the bacterial urease 1082:1101 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 1 9 theme hydrolytic 180:189 arg1 metabolism 191:200 hydrolytic metabolism 180:200 hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics 180:248 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 3 10 theme new 401:403 arg1 antibiotics 405:415 new antibiotics 401:415 new antibiotics that are not hydrolyzed by this enzyme 401:454 They are informative for designing new antibiotics that are not hydrolyzed by this enzyme. 12144777 5 11 theme cilastatin 766:775 arg1 inhibitor 777:785 the therapeutically used cilastatin inhibitor 741:785 the therapeutically used cilastatin inhibitor 741:785 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 1 12 theme Human 81:85 arg1 dipeptidase 93:103 Human renal dipeptidase 81:103 Human renal dipeptidase 81:103 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 1 12 theme Human 81:85 arg1 glycoprotein 125:136 a membrane-bound glycoprotein 108:136 a membrane-bound glycoprotein hydrolyzing dipeptides 108:159 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 6 13 theme bacterial 1086:1094 arg1 urease 1096:1101 the bacterial urease 1082:1101 the bacterial urease 1082:1101 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 14 theme metal 968:972 arg1 ions 974:977 catalytic metal ions 958:977 catalytic metal ions 958:977 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 15 theme unexpected 987:996 arg1 similarities 998:1009 unexpected similarities 987:1009 unexpected similarities to those of the murine adenosine deaminase 987:1052 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 1 16 theme renal 87:91 arg1 dipeptidase 93:103 Human renal dipeptidase 81:103 Human renal dipeptidase 81:103 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 1 16 theme renal 87:91 arg1 glycoprotein 125:136 a membrane-bound glycoprotein 108:136 a membrane-bound glycoprotein hydrolyzing dipeptides 108:159 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 0 17 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human renal dipeptidase 0:43 Crystal structure of human renal dipeptidase involved in beta-lactam hydrolysis. 12144777 4 18 theme binuclear 563:571 arg1 ions 578:581 binuclear zinc ions 563:581 binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel 563:650 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 1 19 theme penem 205:209 arg1 metabolism 191:200 hydrolytic metabolism 180:200 hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics 180:248 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 0 20 theme human 21:25 arg1 dipeptidase 33:43 human renal dipeptidase 21:43 human renal dipeptidase 21:43 Crystal structure of human renal dipeptidase involved in beta-lactam hydrolysis. 12144777 1 21 gly glycoprotein 125:136 arg1 dipeptidase 93:103 Human renal dipeptidase 81:103 Human renal dipeptidase 81:103 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 1 21 gly glycoprotein 125:136 arg1 glycoprotein 125:136 a membrane-bound glycoprotein 108:136 a membrane-bound glycoprotein hydrolyzing dipeptides 108:159 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 2 22 theme inhibitor-liganded 340:357 arg1 structures 263:272 The crystal structures 251:272 The crystal structures of the saccharide-trimmed enzyme 251:305 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 2 22 theme inhibitor-liganded 340:357 arg1 forms 359:363 unliganded and inhibitor-liganded forms 325:363 unliganded and inhibitor-liganded forms 325:363 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 5 23 theme active-site 816:826 arg1 pocket 828:833 the active-site pocket 812:833 the active-site pocket 812:833 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 5 23 theme active-site 816:826 arg1 small 845:849 small 845:849 small 845:849 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 5 24 theme substrates 895:904 arg1 recognition 870:880 precise recognition 862:880 precise recognition of dipeptide substrates 862:904 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 6 25 theme barrel 911:916 arg1 architectures 934:946 The barrel and active-site architectures 907:946 architectures 934:946 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 1 26 theme carbapenem 215:224 arg1 antibiotics 238:248 carbapenem beta-lactam antibiotics 215:248 carbapenem beta-lactam antibiotics 215:248 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 0 27 theme dipeptidase 33:43 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human renal dipeptidase 0:43 Crystal structure of human renal dipeptidase involved in beta-lactam hydrolysis. 12144777 4 28 theme zinc 573:576 arg1 ions 578:581 binuclear zinc ions 563:581 binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel 563:650 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 4 29 theme barrel 645:650 arg1 bottom 631:636 the bottom 627:636 the bottom of the barrel 627:650 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 4 30 from bottom 631:636 arg1 located 616:622 located 616:622 located 616:622 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 1 31 theme beta-lactam 226:236 arg1 antibiotics 238:248 carbapenem beta-lactam antibiotics 215:248 carbapenem beta-lactam antibiotics 215:248 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 0 32 theme renal 27:31 arg1 dipeptidase 33:43 human renal dipeptidase 21:43 human renal dipeptidase 21:43 Crystal structure of human renal dipeptidase involved in beta-lactam hydrolysis. 12144777 4 33 theme molecule 539:546 arg1 subunits 511:518 the (alpha/beta)(8) barrel subunits 484:518 the (alpha/beta)(8) barrel subunits of the homodimeric molecule 484:546 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 5 34 theme inhibitor 777:785 arg1 moiety 731:736 A dipeptidyl moiety 718:736 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor 718:785 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 4 35 theme alpha/beta 489:498 arg1 barrel 504:509 (alpha/beta)(8) barrel 488:509 the (alpha/beta)(8) barrel subunits of the homodimeric molecule 484:546 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 2 36 theme crystal 255:261 arg1 structures 263:272 The crystal structures 251:272 The crystal structures of the saccharide-trimmed enzyme 251:305 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 2 36 theme crystal 255:261 arg1 forms 359:363 unliganded and inhibitor-liganded forms 325:363 unliganded and inhibitor-liganded forms 325:363 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 1 37 theme antibiotics 238:248 arg1 metabolism 191:200 hydrolytic metabolism 180:200 hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics 180:248 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 4 38 theme homodimeric 527:537 arg1 molecule 539:546 the homodimeric molecule 523:546 the homodimeric molecule 523:546 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 1 39 theme membrane-bound 110:123 arg1 dipeptidase 93:103 Human renal dipeptidase 81:103 Human renal dipeptidase 81:103 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 1 39 theme membrane-bound 110:123 arg1 glycoprotein 125:136 a membrane-bound glycoprotein 108:136 a membrane-bound glycoprotein hydrolyzing dipeptides 108:159 Human renal dipeptidase is a membrane-bound glycoprotein hydrolyzing dipeptides and is involved in hydrolytic metabolism of penem and carbapenem beta-lactam antibiotics. 12144777 6 40 theme catalytic 958:966 arg1 ions 974:977 catalytic metal ions 958:977 catalytic metal ions 958:977 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 41 theme active-site 922:932 arg1 architectures 934:946 The barrel and active-site architectures 907:946 architectures 934:946 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 4 42 from site 468:471 arg1 subunits 511:518 the (alpha/beta)(8) barrel subunits 484:518 the (alpha/beta)(8) barrel subunits of the homodimeric molecule 484:546 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 4 42 from site 468:471 arg1 each 476:479 each 476:479 each 476:479 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 4 43 theme active 461:466 arg1 site 468:471 The active site 457:471 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule 457:546 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 2 44 theme enzyme 300:305 arg1 structures 263:272 The crystal structures 251:272 The crystal structures of the saccharide-trimmed enzyme 251:305 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 2 44 theme enzyme 300:305 arg1 forms 359:363 unliganded and inhibitor-liganded forms 325:363 unliganded and inhibitor-liganded forms 325:363 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 4 45 theme tubule 710:715 arg1 membrane 689:696 the microvillar membrane 673:696 the microvillar membrane of a kidney tubule 673:715 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 5 46 theme precise 862:868 arg1 recognition 870:880 precise recognition 862:880 precise recognition of dipeptide substrates 862:904 A dipeptidyl moiety of the therapeutically used cilastatin inhibitor is fully accommodated in the active-site pocket, which is small enough for precise recognition of dipeptide substrates. 12144777 4 47 theme located 616:622 arg1 side-chain 605:614 the Glu125 side-chain 594:614 the Glu125 side-chain located at the bottom of the barrel 594:650 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 2 48 theme saccharide-trimmed 281:298 arg1 enzyme 300:305 the saccharide-trimmed enzyme 277:305 the saccharide-trimmed enzyme 277:305 The crystal structures of the saccharide-trimmed enzyme are determined as unliganded and inhibitor-liganded forms. 12144777 4 49 theme kidney 703:708 arg1 tubule 710:715 a kidney tubule 701:715 a kidney tubule 701:715 The active site in each of the (alpha/beta)(8) barrel subunits of the homodimeric molecule is composed of binuclear zinc ions bridged by the Glu125 side-chain located at the bottom of the barrel, and it faces toward the microvillar membrane of a kidney tubule. 12144777 6 50 theme adenosine 1034:1042 arg1 deaminase 1044:1052 the murine adenosine deaminase 1023:1052 the murine adenosine deaminase 1023:1052 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 6 51 theme murine 1027:1032 arg1 deaminase 1044:1052 the murine adenosine deaminase 1023:1052 the murine adenosine deaminase 1023:1052 The barrel and active-site architectures utilizing catalytic metal ions exhibit unexpected similarities to those of the murine adenosine deaminase and the catalytic domain of the bacterial urease. 12144777 0 52 theme beta-lactam 57:67 arg1 hydrolysis 69:78 beta-lactam hydrolysis 57:78 beta-lactam hydrolysis 57:78 Crystal structure of human renal dipeptidase involved in beta-lactam hydrolysis. 34880232 7 0 theme cryo-EM 875:881 arg1 structure 883:891 the cryo-EM structure 871:891 the cryo-EM structure of system xc- in both the apo and glutamate bound states 871:948 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 8 1 theme human 1137:1141 arg1 cells 1143:1147 human cells 1137:1147 human cells 1137:1147 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 5 2 theme xc- 634:636 arg1 cells 621:625 cancerous cells 611:625 cancerous cells system xc- 611:636 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 7 3 theme bound 937:941 arg1 states 943:948 both the apo and glutamate bound states 910:948 states 943:948 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 8 4 from transport 1124:1132 arg1 cells 1143:1147 human cells 1137:1147 human cells 1137:1147 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 5 5 theme anticancer 700:709 arg1 therapies 711:719 anticancer therapies 700:719 anticancer therapies 700:719 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 4 6 theme cystine 471:477 arg1 xc- 452:454 System xc- 445:454 System xc- 445:454 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 4 6 theme cystine 471:477 arg1 transporter 479:489 a dedicated cystine transporter 459:489 a dedicated cystine transporter 459:489 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 5 7 dep it 672:673 arg1 target 689:694 an attractive target 675:694 it an attractive target for anticancer therapies 672:719 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 4 8 theme System 445:450 arg1 xc- 452:454 System xc- 445:454 System xc- 445:454 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 4 8 theme System 445:450 arg1 transporter 479:489 a dedicated cystine transporter 459:489 a dedicated cystine transporter 459:489 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 5 9 theme reactive 584:591 arg1 species 600:606 reactive oxygen species 584:606 reactive oxygen species 584:606 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 0 10 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis for redox control by the human cystine/glutamate antiporter system 0:81 Molecular basis for redox control by the human cystine/glutamate antiporter system xc. 34880232 5 11 theme species 600:606 arg1 levels 574:579 elevated levels 565:579 elevated levels of reactive oxygen species 565:606 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 5 12 theme oxygen 593:598 arg1 species 600:606 reactive oxygen species 584:606 reactive oxygen species 584:606 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 1 13 theme key 156:158 arg1 constituent 160:170 a key constituent 154:170 a key constituent of the tripeptide glutathione (GSH) 154:206 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 1 13 theme key 156:158 arg1 role 115:118 an essential role 102:118 an essential role 102:118 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 8 14 theme allosteric 984:993 arg1 mechanism 995:1003 an allosteric mechanism 981:1003 an allosteric mechanism for ligand discrimination 981:1029 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 2 15 theme GSH 242:244 arg1 synthesis 246:254 cellular GSH synthesis 233:254 cellular GSH synthesis 233:254 A rate limiting step in cellular GSH synthesis is the availability of cysteine. 34880232 3 16 theme oxidised 346:353 arg1 cystine 366:372 the oxidised di-peptide cystine 342:372 the oxidised di-peptide cystine 342:372 However, circulating cysteine exists in the blood as the oxidised di-peptide cystine, requiring specialised transport systems for its import into the cell. 34880232 5 17 theme cancerous 611:619 arg1 cells 621:625 cancerous cells 611:625 cancerous cells system xc- 611:636 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 0 18 theme antiporter 65:74 arg1 system 76:81 the human cystine/glutamate antiporter system 37:81 the human cystine/glutamate antiporter system 37:81 Molecular basis for redox control by the human cystine/glutamate antiporter system xc. 34880232 7 19 from structure 883:891 arg1 states 943:948 both the apo and glutamate bound states 910:948 states 943:948 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 3 20 theme transport 397:405 arg1 systems 407:413 specialised transport systems 385:413 specialised transport systems 385:413 However, circulating cysteine exists in the blood as the oxidised di-peptide cystine, requiring specialised transport systems for its import into the cell. 34880232 3 21 theme di-peptide 355:364 arg1 cystine 366:372 the oxidised di-peptide cystine 342:372 the oxidised di-peptide cystine 342:372 However, circulating cysteine exists in the blood as the oxidised di-peptide cystine, requiring specialised transport systems for its import into the cell. 34880232 6 22 theme ligand 755:760 arg1 recognition 762:772 ligand recognition 755:772 ligand recognition 755:772 However, the molecular basis for ligand recognition remains elusive, hampering efforts to specifically target this transport system. 34880232 2 23 theme cysteine 279:286 arg1 rate 211:214 A rate 209:214 A rate limiting step in cellular GSH synthesis 209:254 A rate limiting step in cellular GSH synthesis is the availability of cysteine. 34880232 2 23 theme cysteine 279:286 arg1 availability 263:274 the availability 259:274 the availability of cysteine 259:286 A rate limiting step in cellular GSH synthesis is the availability of cysteine. 34880232 2 24 from step 225:228 arg1 synthesis 246:254 cellular GSH synthesis 233:254 cellular GSH synthesis 233:254 A rate limiting step in cellular GSH synthesis is the availability of cysteine. 34880232 8 25 theme ligand 1009:1014 arg1 discrimination 1016:1029 ligand discrimination 1009:1029 ligand discrimination 1009:1029 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 0 26 theme redox 20:24 arg1 control 26:32 redox control 20:32 redox control 20:32 Molecular basis for redox control by the human cystine/glutamate antiporter system xc. 34880232 5 27 theme elevated 565:572 arg1 levels 574:579 elevated levels 565:579 elevated levels of reactive oxygen species 565:606 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 8 28 theme Structural 951:960 arg1 comparisons 962:972 Structural comparisons 951:972 Structural comparisons 951:972 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 1 29 theme tripeptide 179:188 arg1 glutathione 190:200 the tripeptide glutathione 175:200 the tripeptide glutathione (GSH) 175:206 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 1 29 theme tripeptide 179:188 arg1 GSH 203:205 GSH 203:205 GSH 203:205 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 5 30 theme attractive 678:687 arg1 target 689:694 an attractive target 675:694 it an attractive target for anticancer therapies 672:719 To counteract elevated levels of reactive oxygen species in cancerous cells system xc- is frequently upregulated, making it an attractive target for anticancer therapies. 34880232 7 31 theme glutamate 927:935 arg1 states 943:948 both the apo and glutamate bound states 910:948 states 943:948 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 1 32 theme glutathione 190:200 arg1 constituent 160:170 a key constituent 154:170 a key constituent of the tripeptide glutathione (GSH) 154:206 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 1 32 theme glutathione 190:200 arg1 role 115:118 an essential role 102:118 an essential role 102:118 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 8 33 theme molecular 1045:1053 arg1 dynamics 1055:1062 molecular dynamics 1045:1062 molecular dynamics 1045:1062 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 4 34 theme intracellular 526:538 arg1 glutamate 540:548 intracellular glutamate 526:548 intracellular glutamate 526:548 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 8 35 theme cell-based 1068:1077 arg1 assays 1079:1084 cell-based assays 1068:1084 cell-based assays 1068:1084 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 3 36 theme specialised 385:395 arg1 systems 407:413 specialised transport systems 385:413 specialised transport systems 385:413 However, circulating cysteine exists in the blood as the oxidised di-peptide cystine, requiring specialised transport systems for its import into the cell. 34880232 4 37 from cystine 502:508 arg1 exchange 513:520 exchange 513:520 exchange 513:520 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 0 38 theme human 41:45 arg1 system 76:81 the human cystine/glutamate antiporter system 37:81 the human cystine/glutamate antiporter system 37:81 Molecular basis for redox control by the human cystine/glutamate antiporter system xc. 34880232 6 39 theme transport 837:845 arg1 system 847:852 this transport system 832:852 this transport system 832:852 However, the molecular basis for ligand recognition remains elusive, hampering efforts to specifically target this transport system. 34880232 6 40 theme molecular 735:743 arg1 basis 745:749 the molecular basis 731:749 the molecular basis for ligand recognition 731:772 However, the molecular basis for ligand recognition remains elusive, hampering efforts to specifically target this transport system. 34880232 1 41 theme essential 105:113 arg1 role 115:118 an essential role 102:118 an essential role 102:118 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 1 41 theme essential 105:113 arg1 constituent 160:170 a key constituent 154:170 a key constituent of the tripeptide glutathione (GSH) 154:206 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 7 42 theme apo 919:921 arg1 states 943:948 both the apo and glutamate bound states 910:948 states 943:948 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 7 43 theme system 896:901 arg1 xc- 903:905 system xc- 896:905 system xc- 896:905 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 8 44 theme cystine 1116:1122 arg1 transport 1124:1132 cystine transport 1116:1132 cystine transport in human cells 1116:1147 Structural comparisons reveal an allosteric mechanism for ligand discrimination, supported by molecular dynamics and cell-based assays, establishing a mechanism for cystine transport in human cells. 34880232 2 45 theme cellular 233:240 arg1 synthesis 246:254 cellular GSH synthesis 233:254 cellular GSH synthesis 233:254 A rate limiting step in cellular GSH synthesis is the availability of cysteine. 34880232 7 46 theme xc- 903:905 arg1 structure 883:891 the cryo-EM structure 871:891 the cryo-EM structure of system xc- in both the apo and glutamate bound states 871:948 Here we present the cryo-EM structure of system xc- in both the apo and glutamate bound states. 34880232 1 47 theme cellular 123:130 arg1 homoeostasis 138:149 cellular redox homoeostasis 123:149 cellular redox homoeostasis 123:149 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 3 48 theme circulating 298:308 arg1 cysteine 310:317 circulating cysteine 298:317 circulating cysteine 298:317 However, circulating cysteine exists in the blood as the oxidised di-peptide cystine, requiring specialised transport systems for its import into the cell. 34880232 1 49 theme redox 132:136 arg1 homoeostasis 138:149 cellular redox homoeostasis 123:149 cellular redox homoeostasis 123:149 Cysteine plays an essential role in cellular redox homoeostasis as a key constituent of the tripeptide glutathione (GSH). 34880232 0 50 theme cystine/glutamate 47:63 arg1 system 76:81 the human cystine/glutamate antiporter system 37:81 the human cystine/glutamate antiporter system 37:81 Molecular basis for redox control by the human cystine/glutamate antiporter system xc. 34880232 4 51 theme dedicated 461:469 arg1 xc- 452:454 System xc- 445:454 System xc- 445:454 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 34880232 4 51 theme dedicated 461:469 arg1 transporter 479:489 a dedicated cystine transporter 459:489 a dedicated cystine transporter 459:489 System xc- is a dedicated cystine transporter, importing cystine in exchange for intracellular glutamate. 30867591 6 0 theme LAT1-4F2hc 1174:1183 arg1 structures 1154:1163 the cryo-electron microscopy structures 1125:1163 the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1125:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 3 1 theme cells 544:548 arg1 range 528:532 a wide range 521:532 a wide range of tumour cells 521:548 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 6 2 from resolutions 1264:1274 arg1 acid 1256:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid 1211:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1211:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 4 3 theme acid 643:646 arg1 complex 660:666 a heteromeric amino acid transporter complex 623:666 a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 623:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 10 4 theme LAT1-4F2hc 1679:1688 arg1 complex 1690:1696 the LAT1-4F2hc complex 1675:1696 the LAT1-4F2hc complex 1675:1696 Together, our characterizations shed light on the architecture of the LAT1-4F2hc complex, and provide insights into its function and the mechanisms through which it might be associated with disease. 30867591 6 5 theme microscopy 1143:1152 arg1 structures 1154:1163 the cryo-electron microscopy structures 1125:1163 the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1125:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 1 6 theme cross-membrane 157:170 arg1 flux 172:175 the cross-membrane flux 153:175 the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3 153:246 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 5 7 theme extensive 876:884 arg1 characterization 897:912 extensive cell-based characterization 876:912 extensive cell-based characterization of the LAT1-4F2hc complex 876:938 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 6 8 theme 2-amino-2-norbornanecarboxylic 1225:1254 arg1 acid 1256:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid 1211:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1211:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 4 9 theme heteromeric 625:635 arg1 complex 660:666 a heteromeric amino acid transporter complex 623:666 a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 623:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 1 10 theme neutral 186:192 arg1 acids 200:204 large neutral amino acids 180:204 large neutral amino acids 180:204 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 4 11 theme plasma 848:853 arg1 membrane8,9 855:865 the plasma membrane8,9 844:865 the plasma membrane8,9 844:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 4 12 theme chain 704:708 arg1 glycoprotein 759:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein 673:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 1 13 from flux 172:175 arg1 manner1-3 238:246 a sodium- and pH-independent manner1-3 209:246 a sodium- and pH-independent manner1-3 209:246 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 4 14 dep known 723:727 arg1 also 718:721 also 718:721 also 718:721 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 5 15 theme LAT1-4F2hc 921:930 arg1 complex 932:938 the LAT1-4F2hc complex 917:938 the LAT1-4F2hc complex 917:938 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 6 16 theme Å 1293:1293 arg1 resolutions 1264:1274 resolutions 1264:1274 resolutions of 3.3 Å and 3.5 Å, respectively 1264:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 7 17 theme open 1334:1337 arg1 conformation 1339:1350 an inward open conformation 1324:1350 an inward open conformation 1324:1350 LAT1 exhibits an inward open conformation. 30867591 4 18 theme 4F2 673:675 arg1 chain 704:708 4F2 cell-surface antigen heavy chain 673:708 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 0 19 theme transporter 57:67 arg1 complex 69:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 3 20 theme anti-cancer 589:599 arg1 drugs7-11 601:609 anti-cancer drugs7-11 589:609 anti-cancer drugs7-11 589:609 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 4 21 theme antigen 690:696 arg1 chain 704:708 4F2 cell-surface antigen heavy chain 673:708 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 9 22 theme transport 1574:1582 arg1 activity 1584:1591 the transport activity 1570:1591 the transport activity of the complex 1570:1606 Biochemical analysis reveals that 4F2hc is essential for the transport activity of the complex. 30867591 5 23 theme structural 944:953 arg1 determination 955:967 structural determination 944:967 structural determination of its homologues in bacteria 944:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 4 24 theme type 742:745 arg1 glycoprotein 759:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein 673:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 6 25 from structures 1154:1163 arg1 complex 1198:1204 complex 1198:1204 complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1198:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 2 26 theme pharmaceutical 374:387 arg1 L-3,4-dihydroxyphenylalanine 427:454 L-3,4-dihydroxyphenylalanine 427:454 L-3,4-dihydroxyphenylalanine across membranes2-6 427:474 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 2 26 theme pharmaceutical 374:387 arg1 drugs 389:393 pharmaceutical drugs 374:393 pharmaceutical drugs 374:393 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 7 27 theme inward 1327:1332 arg1 conformation 1339:1350 an inward open conformation 1324:1350 an inward open conformation 1324:1350 LAT1 exhibits an inward open conformation. 30867591 8 28 theme disulfide 1363:1371 arg1 association 1378:1388 a disulfide bond association 1361:1388 a disulfide bond association 1361:1388 Besides a disulfide bond association, LAT1 also interacts extensively with 4F2hc on the extracellular side, within the membrane, and on the intracellular side. 30867591 2 29 theme hormones 364:371 arg1 permeation 342:351 the permeation 338:351 the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6 338:474 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 1 30 theme amino 89:93 arg1 transporter 100:110 The L-type amino acid transporter 1 78:112 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) 78:141 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 2 31 theme hormone 400:406 arg1 precursors 408:417 hormone precursors 400:417 hormone precursors 400:417 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 2 31 theme hormone 400:406 arg1 L-3,4-dihydroxyphenylalanine 427:454 L-3,4-dihydroxyphenylalanine 427:454 L-3,4-dihydroxyphenylalanine across membranes2-6 427:474 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 5 32 theme complex 932:938 arg1 determination 955:967 structural determination 944:967 structural determination of its homologues in bacteria 944:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 5 32 theme complex 932:938 arg1 characterization 897:912 extensive cell-based characterization 876:912 extensive cell-based characterization of the LAT1-4F2hc complex 876:938 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 0 33 theme LAT1-4F2hc 23:32 arg1 complex 69:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 0 34 theme amino 46:50 arg1 complex 69:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 9 35 theme complex 1600:1606 arg1 activity 1584:1591 the transport activity 1570:1591 the transport activity of the complex 1570:1606 Biochemical analysis reveals that 4F2hc is essential for the transport activity of the complex. 30867591 4 36 theme membrane 750:757 arg1 glycoprotein 759:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein 673:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 2 37 theme superfamily 310:320 arg1 LAT1 249:252 LAT1 249:252 LAT1 249:252 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 2 37 theme superfamily 310:320 arg1 antiporter 258:267 an antiporter 255:267 an antiporter of the amino acid-polyamine-organocation superfamily 255:320 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 1 38 theme amino 194:198 arg1 acids 200:204 large neutral amino acids 180:204 large neutral amino acids 180:204 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 5 39 theme homologues 976:985 arg1 determination 955:967 structural determination 944:967 structural determination of its homologues in bacteria 944:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 5 39 theme homologues 976:985 arg1 characterization 897:912 extensive cell-based characterization 876:912 extensive cell-based characterization of the LAT1-4F2hc complex 876:938 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 0 40 theme acid 52:55 arg1 complex 69:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 1 41 theme sodium- 211:217 arg1 manner1-3 238:246 a sodium- and pH-independent manner1-3 209:246 a sodium- and pH-independent manner1-3 209:246 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 6 42 theme Å 1283:1283 arg1 resolutions 1264:1274 resolutions 1264:1274 resolutions of 3.3 Å and 3.5 Å, respectively 1264:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 3 43 theme tumour 537:542 arg1 cells 544:548 tumour cells 537:548 tumour cells 537:548 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 1 44 theme pH-independent 223:236 arg1 manner1-3 238:246 a sodium- and pH-independent manner1-3 209:246 a sodium- and pH-independent manner1-3 209:246 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 6 45 theme human 1168:1172 arg1 LAT1-4F2hc 1174:1183 human LAT1-4F2hc 1168:1183 human LAT1-4F2hc alone 1168:1189 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 4 46 dep chain 704:708 arg1 known 723:727 known 723:727 known as SLC3A2 723:737 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 5 47 from bacteria 990:997 arg1 determination 955:967 structural determination 944:967 structural determination of its homologues in bacteria 944:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 5 47 from bacteria 990:997 arg1 characterization 897:912 extensive cell-based characterization 876:912 extensive cell-based characterization of the LAT1-4F2hc complex 876:938 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 4 48 theme amino 637:641 arg1 complex 660:666 a heteromeric amino acid transporter complex 623:666 a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 623:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 2 49 theme amino 276:280 arg1 superfamily 310:320 the amino acid-polyamine-organocation superfamily 272:320 the amino acid-polyamine-organocation superfamily 272:320 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 5 50 from homologues 976:985 arg1 bacteria 990:997 bacteria 990:997 bacteria 990:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 4 51 theme LAT1 811:814 arg1 stability 798:806 the stability 794:806 the stability of LAT1 794:814 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 6 52 theme cryo-electron 1129:1141 arg1 microscopy 1143:1152 cryo-electron microscopy 1129:1152 the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1125:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 4 53 theme transporter 648:658 arg1 complex 660:666 a heteromeric amino acid transporter complex 623:666 a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 623:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 8 54 from 4F2hc 1428:1432 arg1 side 1455:1458 the extracellular side 1437:1458 the extracellular side 1437:1458 Besides a disulfide bond association, LAT1 also interacts extensively with 4F2hc on the extracellular side, within the membrane, and on the intracellular side. 30867591 8 54 from 4F2hc 1428:1432 arg1 side 1507:1510 the intracellular side 1489:1510 the intracellular side 1489:1510 Besides a disulfide bond association, LAT1 also interacts extensively with 4F2hc on the extracellular side, within the membrane, and on the intracellular side. 30867591 8 55 theme extracellular 1441:1453 arg1 side 1455:1458 the extracellular side 1437:1458 the extracellular side 1437:1458 Besides a disulfide bond association, LAT1 also interacts extensively with 4F2hc on the extracellular side, within the membrane, and on the intracellular side. 30867591 5 56 theme working 1048:1054 arg1 mechanism 1056:1064 the working mechanism 1044:1064 the working mechanism of the complex 1044:1079 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 3 57 theme potential 568:576 arg1 target 578:583 a potential target 566:583 a potential target for anti-cancer drugs7-11 566:609 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 3 57 theme potential 568:576 arg1 it 555:556 it 555:556 it 555:556 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 1 58 theme large 180:184 arg1 acids 200:204 large neutral amino acids 180:204 large neutral amino acids 180:204 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 6 59 theme inhibitor 1215:1223 arg1 acid 1256:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid 1211:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1211:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 4 60 theme heavy 698:702 arg1 chain 704:708 4F2 cell-surface antigen heavy chain 673:708 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 3 61 located observed 509:516 arg2 Overexpression 477:490 Overexpression 477:490 Overexpression of LAT1 477:498 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 3 61 located observed 509:516 arg1 range 528:532 a wide range 521:532 a wide range of tumour cells 521:548 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 5 62 theme cell-based 886:895 arg1 characterization 897:912 extensive cell-based characterization 876:912 extensive cell-based characterization of the LAT1-4F2hc complex 876:938 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 10 63 theme complex 1690:1696 arg1 architecture 1659:1670 the architecture 1655:1670 the architecture of the LAT1-4F2hc complex 1655:1696 Together, our characterizations shed light on the architecture of the LAT1-4F2hc complex, and provide insights into its function and the mechanisms through which it might be associated with disease. 30867591 0 64 theme complex 69:75 arg1 Structure 0:8 Structure 0:8 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 0:76 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 1 65 dep transporter 100:110 arg1 known 126:130 known 126:130 known as SLC7A5 126:140 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 9 66 theme Biochemical 1513:1523 arg1 analysis 1525:1532 Biochemical analysis 1513:1532 Biochemical analysis 1513:1532 Biochemical analysis reveals that 4F2hc is essential for the transport activity of the complex. 30867591 8 67 theme intracellular 1493:1505 arg1 side 1507:1510 the intracellular side 1489:1510 the intracellular side 1489:1510 Besides a disulfide bond association, LAT1 also interacts extensively with 4F2hc on the extracellular side, within the membrane, and on the intracellular side. 30867591 4 68 theme cell-surface 677:688 arg1 chain 704:708 4F2 cell-surface antigen heavy chain 673:708 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 6 69 with complex 1198:1204 arg1 acid 1256:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid 1211:1259 the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively 1211:1307 Here we report the cryo-electron microscopy structures of human LAT1-4F2hc alone and in complex with the inhibitor 2-amino-2-norbornanecarboxylic acid at resolutions of 3.3 Å and 3.5 Å, respectively. 30867591 2 70 theme acid-polyamine-organocation 282:308 arg1 superfamily 310:320 the amino acid-polyamine-organocation superfamily 272:320 the amino acid-polyamine-organocation superfamily 272:320 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 5 71 theme complex 1073:1079 arg1 mechanism 1056:1064 the working mechanism 1044:1064 the working mechanism of the complex 1044:1079 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 5 71 theme complex 1073:1079 arg1 interactions 1004:1015 the interactions 1000:1015 the interactions between LAT1 and 4F2hc 1000:1038 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 4 72 theme -a 739:740 arg1 glycoprotein 759:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein 673:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 5 73 from characterization 897:912 arg1 bacteria 990:997 bacteria 990:997 bacteria 990:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 30867591 1 74 theme L-type 82:87 arg1 transporter 100:110 The L-type amino acid transporter 1 78:112 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) 78:141 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 2 75 theme thyroid 356:362 arg1 L-3,4-dihydroxyphenylalanine 427:454 L-3,4-dihydroxyphenylalanine 427:454 L-3,4-dihydroxyphenylalanine across membranes2-6 427:474 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 2 75 theme thyroid 356:362 arg1 hormones 364:371 thyroid hormones 356:371 thyroid hormones 356:371 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 0 76 theme human 17:21 arg1 complex 69:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 1 77 theme acid 95:98 arg1 transporter 100:110 The L-type amino acid transporter 1 78:112 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) 78:141 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 2 78 theme precursors 408:417 arg1 permeation 342:351 the permeation 338:351 the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6 338:474 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 4 79 with complex 660:666 arg1 glycoprotein 759:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein 673:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 8 80 theme bond 1373:1376 arg1 association 1378:1388 a disulfide bond association 1361:1388 a disulfide bond association 1361:1388 Besides a disulfide bond association, LAT1 also interacts extensively with 4F2hc on the extracellular side, within the membrane, and on the intracellular side. 30867591 0 81 theme heteromeric 34:44 arg1 complex 69:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 the human LAT1-4F2hc heteromeric amino acid transporter complex 13:75 Structure of the human LAT1-4F2hc heteromeric amino acid transporter complex. 30867591 2 82 theme drugs 389:393 arg1 permeation 342:351 the permeation 338:351 the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6 338:474 LAT1, an antiporter of the amino acid-polyamine-organocation superfamily, also catalyses the permeation of thyroid hormones, pharmaceutical drugs, and hormone precursors such as L-3,4-dihydroxyphenylalanine across membranes2-6. 30867591 4 83 gly glycoprotein 759:770 arg1 glycoprotein 759:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein 673:770 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9 673:865 LAT1 forms a heteromeric amino acid transporter complex with 4F2 cell-surface antigen heavy chain (4F2hc; also known as SLC3A2)-a type II membrane glycoprotein that is essential for the stability of LAT1 and for its localization to the plasma membrane8,9. 30867591 1 84 dep known 126:130 arg1 also 121:124 also 121:124 also 121:124 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 3 85 theme LAT1 495:498 arg1 Overexpression 477:490 Overexpression 477:490 Overexpression of LAT1 477:498 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 1 86 theme acids 200:204 arg1 flux 172:175 the cross-membrane flux 153:175 the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3 153:246 The L-type amino acid transporter 1 (LAT1; also known as SLC7A5) catalyses the cross-membrane flux of large neutral amino acids in a sodium- and pH-independent manner1-3. 30867591 3 87 theme wide 523:526 arg1 range 528:532 a wide range 521:532 a wide range of tumour cells 521:548 Overexpression of LAT1 has been observed in a wide range of tumour cells, and it is thus a potential target for anti-cancer drugs7-11. 30867591 5 88 from determination 955:967 arg1 bacteria 990:997 bacteria 990:997 bacteria 990:997 Despite extensive cell-based characterization of the LAT1-4F2hc complex and structural determination of its homologues in bacteria, the interactions between LAT1 and 4F2hc and the working mechanism of the complex remain largely unknown12-19. 33203861 7 0 theme regulation 1019:1028 arg1 understanding 996:1008 the mechanistic understanding 980:1008 the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases 980:1068 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 3 1 theme overall 447:453 arg1 resolution 455:464 an overall resolution 444:464 an overall resolution of 3.1 angstroms 444:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 1 2 theme neuronal 213:220 arg1 excitability 222:233 neuronal excitability 213:233 neuronal excitability 213:233 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 5 3 theme Ca2+ 647:650 arg1 site 660:663 a Ca2+ binding site 645:663 a Ca2+ binding site in Cav channels 645:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 7 4 theme NALCN 1013:1017 arg1 regulation 1019:1028 NALCN regulation 1013:1028 NALCN regulation 1013:1028 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 1 5 theme central 127:133 arg1 system 143:148 the central nervous system 123:148 the central nervous system 123:148 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 4 6 theme extracellular 530:542 arg1 loops 544:548 the extracellular loops 526:548 the extracellular loops of NALCN 526:557 FAM155A forms extensive interactions with the extracellular loops of NALCN that may help stabilize NALCN in the membrane. 33203861 5 7 theme Na+ 608:610 arg1 reminiscent 630:640 reminiscent 630:640 reminiscent of a Ca2+ binding site in Cav channels 630:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 5 7 theme Na+ 608:610 arg1 site 624:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 3 8 from resolution 455:464 arg1 structure 392:400 the cryo-EM structure 380:400 the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms 380:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 3 8 from resolution 455:464 arg1 complex 420:426 complex 420:426 complex with FAM155A at an overall resolution of 3.1 angstroms 420:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 2 9 theme channelosome 262:273 arg1 Dysfunctions 236:247 Dysfunctions 236:247 Dysfunctions 236:247 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 4 10 theme NALCN 553:557 arg1 loops 544:548 the extracellular loops 526:548 the extracellular loops of NALCN 526:557 FAM155A forms extensive interactions with the extracellular loops of NALCN that may help stabilize NALCN in the membrane. 33203861 5 11 theme Cav 668:670 arg1 channels 672:679 Cav channels 668:679 Cav channels 668:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 5 12 theme ion-binding 612:622 arg1 reminiscent 630:640 reminiscent 630:640 reminiscent of a Ca2+ binding site in Cav channels 630:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 5 12 theme ion-binding 612:622 arg1 site 624:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 0 13 with complex 52:58 arg1 FAM155A 65:71 FAM155A 65:71 FAM155A 65:71 Structure of the human sodium leak channel NALCN in complex with FAM155A. 33203861 2 14 theme NALCN 256:260 arg1 channelosome 262:273 the NALCN channelosome 252:273 the NALCN channelosome 252:273 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 6 15 theme intracellular 793:805 arg1 gate 807:810 the intracellular gate 789:810 the intracellular gate 789:810 Despite its 'leaky' nature, the channel is closed and the intracellular gate is sealed by S6I, II-III linker and III-IV linker. 33203861 7 16 theme mechanistic 984:994 arg1 understanding 996:1008 the mechanistic understanding 980:1008 the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases 980:1068 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 3 17 theme angstroms 473:481 arg1 resolution 455:464 an overall resolution 444:464 an overall resolution of 3.1 angstroms 444:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 1 18 theme nervous 135:141 arg1 system 143:148 the central nervous system 123:148 the central nervous system 123:148 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 7 19 theme channelosome-related 1040:1059 arg1 diseases 1061:1068 NALCN channelosome-related diseases 1034:1068 NALCN channelosome-related diseases 1034:1068 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 5 20 theme unique 703:708 arg1 filter 727:732 the unique EEKE selectivity filter 699:732 the unique EEKE selectivity filter 699:732 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 2 21 theme diseases 354:361 arg1 diseases 354:361 human diseases 348:361 human diseases 348:361 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 2 21 theme diseases 354:361 arg1 variety 337:343 a variety 335:343 a variety of human diseases 335:361 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 2 22 theme several 287:293 arg1 subunits 305:312 several auxiliary subunits 287:312 several auxiliary subunits 287:312 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 2 23 theme human 348:352 arg1 diseases 354:361 human diseases 348:361 human diseases 348:361 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 6 24 theme leaky 748:752 arg1 nature 755:760 its 'leaky' nature 743:760 its 'leaky' nature 743:760 Despite its 'leaky' nature, the channel is closed and the intracellular gate is sealed by S6I, II-III linker and III-IV linker. 33203861 3 25 theme cryo-EM 384:390 arg1 structure 392:400 the cryo-EM structure 380:400 the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms 380:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 0 26 theme human 17:21 arg1 channel 35:41 the human sodium leak channel 13:41 the human sodium leak channel 13:41 Structure of the human sodium leak channel NALCN in complex with FAM155A. 33203861 4 27 theme extensive 498:506 arg1 interactions 508:519 extensive interactions 498:519 extensive interactions with the extracellular loops of NALCN that may help stabilize NALCN in the membrane 498:603 FAM155A forms extensive interactions with the extracellular loops of NALCN that may help stabilize NALCN in the membrane. 33203861 7 28 theme important 961:969 arg1 clues 971:975 important clues 961:975 important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases 961:1068 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 4 29 with interactions 508:519 arg1 loops 544:548 the extracellular loops 526:548 the extracellular loops of NALCN 526:557 FAM155A forms extensive interactions with the extracellular loops of NALCN that may help stabilize NALCN in the membrane. 33203861 5 30 from channels 672:679 arg1 reminiscent 630:640 reminiscent 630:640 reminiscent of a Ca2+ binding site in Cav channels 630:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 5 30 from channels 672:679 arg1 site 624:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 7 31 theme diseases 1061:1068 arg1 understanding 996:1008 the mechanistic understanding 980:1008 the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases 980:1068 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 0 32 theme leak 30:33 arg1 channel 35:41 the human sodium leak channel 13:41 the human sodium leak channel 13:41 Structure of the human sodium leak channel NALCN in complex with FAM155A. 33203861 6 33 theme III-IV 848:853 arg1 linker 855:860 III-IV linker 848:860 III-IV linker 848:860 Despite its 'leaky' nature, the channel is closed and the intracellular gate is sealed by S6I, II-III linker and III-IV linker. 33203861 5 34 theme binding 652:658 arg1 site 660:663 a Ca2+ binding site 645:663 a Ca2+ binding site in Cav channels 645:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 5 35 theme site 660:663 arg1 reminiscent 630:640 reminiscent 630:640 reminiscent of a Ca2+ binding site in Cav channels 630:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 5 35 theme site 660:663 arg1 site 624:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site 606:627 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 4 36 from NALCN 583:587 arg1 membrane 596:603 the membrane 592:603 the membrane 592:603 FAM155A forms extensive interactions with the extracellular loops of NALCN that may help stabilize NALCN in the membrane. 33203861 3 37 theme human 405:409 arg1 NALCN 411:415 human NALCN 405:415 human NALCN 405:415 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 0 38 theme sodium 23:28 arg1 channel 35:41 the human sodium leak channel 13:41 the human sodium leak channel 13:41 Structure of the human sodium leak channel NALCN in complex with FAM155A. 33203861 3 39 with complex 420:426 arg1 FAM155A 433:439 FAM155A 433:439 FAM155A 433:439 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 7 40 theme Na+ 908:910 arg1 permeation 912:921 Na+ permeation 908:921 Na+ permeation 908:921 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 1 41 theme resting 174:180 arg1 permeability 186:197 the resting Na+ permeability 170:197 the resting Na+ permeability that controls neuronal excitability 170:233 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 5 42 theme selectivity 715:725 arg1 filter 727:732 the unique EEKE selectivity filter 699:732 the unique EEKE selectivity filter 699:732 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 7 43 theme sensitivity 935:945 arg1 basis 899:903 the molecular basis 885:903 the molecular basis of Na+ permeation and voltage sensitivity 885:945 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 7 44 theme permeation 912:921 arg1 basis 899:903 the molecular basis 885:903 the molecular basis of Na+ permeation and voltage sensitivity 885:945 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 1 45 theme sodium 83:88 arg1 channel 95:101 a sodium leak channel 81:101 a sodium leak channel expressed mainly in the central nervous system 81:148 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 1 45 theme sodium 83:88 arg1 NALCN 74:78 NALCN 74:78 NALCN 74:78 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 0 46 theme channel 35:41 arg1 Structure 0:8 Structure 0:8 Structure of the human sodium leak channel 0:41 Structure of the human sodium leak channel NALCN in complex with FAM155A. 33203861 6 47 theme II-III 830:835 arg1 linker 837:842 II-III linker 830:842 II-III linker 830:842 Despite its 'leaky' nature, the channel is closed and the intracellular gate is sealed by S6I, II-III linker and III-IV linker. 33203861 1 48 theme leak 90:93 arg1 channel 95:101 a sodium leak channel 81:101 a sodium leak channel expressed mainly in the central nervous system 81:148 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 1 48 theme leak 90:93 arg1 NALCN 74:78 NALCN 74:78 NALCN 74:78 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 1 49 theme Na+ 182:184 arg1 permeability 186:197 the resting Na+ permeability 170:197 the resting Na+ permeability that controls neuronal excitability 170:233 NALCN, a sodium leak channel expressed mainly in the central nervous system, is responsible for the resting Na+ permeability that controls neuronal excitability. 33203861 7 50 theme voltage 927:933 arg1 sensitivity 935:945 voltage sensitivity 927:945 voltage sensitivity 927:945 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 5 51 from site 660:663 arg1 channels 672:679 Cav channels 668:679 Cav channels 668:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 2 52 theme auxiliary 295:303 arg1 subunits 305:312 several auxiliary subunits 287:312 several auxiliary subunits 287:312 Dysfunctions of the NALCN channelosome, NALCN with several auxiliary subunits, are associated with a variety of human diseases. 33203861 3 53 theme NALCN 411:415 arg1 structure 392:400 the cryo-EM structure 380:400 the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms 380:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 7 54 theme molecular 889:897 arg1 basis 899:903 the molecular basis 885:903 the molecular basis of Na+ permeation and voltage sensitivity 885:945 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 7 55 theme NALCN 1034:1038 arg1 diseases 1061:1068 NALCN channelosome-related diseases 1034:1068 NALCN channelosome-related diseases 1034:1068 Our study establishes the molecular basis of Na+ permeation and voltage sensitivity, and provides important clues to the mechanistic understanding of NALCN regulation and NALCN channelosome-related diseases. 33203861 5 56 theme EEKE 710:713 arg1 filter 727:732 the unique EEKE selectivity filter 699:732 the unique EEKE selectivity filter 699:732 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 33203861 3 57 from structure 392:400 arg1 resolution 455:464 an overall resolution 444:464 an overall resolution of 3.1 angstroms 444:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 3 57 from structure 392:400 arg1 complex 420:426 complex 420:426 complex with FAM155A at an overall resolution of 3.1 angstroms 420:481 Here, we report the cryo-EM structure of human NALCN in complex with FAM155A at an overall resolution of 3.1 angstroms. 33203861 5 58 from reminiscent 630:640 arg1 channels 672:679 Cav channels 668:679 Cav channels 668:679 A Na+ ion-binding site, reminiscent of a Ca2+ binding site in Cav channels, is identified in the unique EEKE selectivity filter. 6371807 5 0 theme oligosaccharides 779:794 arg1 three 754:758 three 754:758 three 754:758 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 5 0 theme oligosaccharides 779:794 arg1 oligosaccharides 779:794 the glucosamine oligosaccharides 763:794 the glucosamine oligosaccharides 763:794 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 6 1 theme glucosamine 925:935 arg1 oligosaccharides 937:952 the five glucosamine oligosaccharides 916:952 the five glucosamine oligosaccharides 916:952 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 7 2 theme residues 1136:1143 arg1 Clusters 1113:1120 Clusters 1113:1120 Clusters of tryptophan residues 1113:1143 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 6 3 theme hemopexin 1102:1110 arg1 surface 1091:1097 the surface 1087:1097 the surface of hemopexin 1087:1110 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 9 4 theme human 1433:1437 arg1 hemopexin 1439:1447 human hemopexin 1433:1447 human hemopexin 1433:1447 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 0 5 theme unusual 66:72 arg1 clustering 74:83 unusual clustering 66:83 unusual clustering of tryptophan residues 66:106 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 4 6 theme amino-terminal 498:511 arg1 O-linked 541:548 O-linked 541:548 O-linked 541:548 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 6 theme amino-terminal 498:511 arg1 residue 523:529 the amino-terminal threonine residue 494:529 the amino-terminal threonine residue 494:529 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 5 7 theme galactosamine 646:658 arg1 oligosaccharide 660:674 The galactosamine oligosaccharide 642:674 The galactosamine oligosaccharide 642:674 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 5 7 theme galactosamine 646:658 arg1 located 716:722 located 716:722 located 716:722 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 7 8 theme tryptophan 1125:1134 arg1 residues 1136:1143 tryptophan residues 1125:1143 tryptophan residues 1125:1143 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 1 9 theme enzymatic 241:249 arg1 digests 251:257 chemical and enzymatic digests 228:257 chemical and enzymatic digests of the protein 228:272 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 0 10 theme tryptophan 88:97 arg1 residues 99:106 tryptophan residues 88:106 tryptophan residues 88:106 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 9 11 theme heme-binding 1633:1644 arg1 proteins 1646:1653 other heme-binding proteins 1627:1653 other heme-binding proteins 1627:1653 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 2 12 theme Human 275:279 arg1 hemopexin 281:289 Human hemopexin 275:289 Human hemopexin 275:289 Human hemopexin consists of about 440 amino acid residues. 6371807 5 13 from located 716:722 arg1 region 746:751 the amino-terminal region 727:751 the amino-terminal region 727:751 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 7 14 theme tryptophan 1205:1214 arg1 residues 1216:1223 three or four tryptophan residues 1191:1223 three or four tryptophan residues separated by 0-12 other residues 1191:1256 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 6 15 theme middle 1002:1007 arg1 region 1009:1014 the middle region 998:1014 the middle region of the protein 998:1029 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 16 from surface 1091:1097 arg1 beta-turns 1062:1071 beta-turns 1062:1071 beta-turns 1062:1071 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 4 17 attach O-linked 541:548 arg2 O-linked 541:548 O-linked 541:548 O-linked 541:548 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 17 attach O-linked 541:548 arg1 oligosaccharide 569:583 a galactosamine oligosaccharide 553:583 a galactosamine oligosaccharide that has not previously been identified in this protein 553:639 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 17 attach O-linked 541:548 arg2 residue 523:529 the amino-terminal threonine residue 494:529 the amino-terminal threonine residue 494:529 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 6 18 theme protein 1023:1029 arg1 region 1009:1014 the middle region 998:1014 the middle region of the protein 998:1029 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 5 19 located located 716:722 arg1 region 746:751 the amino-terminal region 727:751 the amino-terminal region 727:751 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 5 19 located located 716:722 arg2 oligosaccharide 660:674 The galactosamine oligosaccharide 642:674 The galactosamine oligosaccharide 642:674 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 5 19 located located 716:722 arg2 located 716:722 located 716:722 located 716:722 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 5 19 located located 716:722 arg2 oligosaccharide 696:710 one glucosamine oligosaccharide 680:710 one glucosamine oligosaccharide 680:710 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 8 20 theme heme 1367:1370 arg1 binding 1356:1362 the binding 1352:1362 the binding of heme 1352:1370 This clustering is significant because both histidine and tryptophan have been implicated in the binding of heme. 6371807 1 21 theme sequence 172:179 arg1 analysis 181:188 sequence analysis 172:188 sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein 172:272 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 5 22 theme carboxyl-terminal 868:884 arg1 region 886:891 the carboxyl-terminal region 864:891 the carboxyl-terminal region of the protein 864:906 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 0 23 theme residues 99:106 arg1 sites 56:60 O-glycosyl and N-glycosyl sites 30:60 O-glycosyl and N-glycosyl sites 30:60 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 0 23 theme residues 99:106 arg1 clustering 74:83 unusual clustering 66:83 unusual clustering of tryptophan residues 66:106 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 0 23 theme residues 99:106 arg1 residues 99:106 tryptophan residues 88:106 tryptophan residues 88:106 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 4 24 theme virtual 474:480 arg1 feature 459:465 A unique structural feature 439:465 A unique structural feature 439:465 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 24 theme virtual 474:480 arg1 blocking 482:489 the virtual blocking 470:489 the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein 470:639 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 6 25 dep flank 1056:1060 arg1 beta-turns 1062:1071 beta-turns 1062:1071 beta-turns 1062:1071 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 5 26 from region 746:751 arg1 oligosaccharide 660:674 The galactosamine oligosaccharide 642:674 The galactosamine oligosaccharide 642:674 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 5 26 from region 746:751 arg1 located 716:722 located 716:722 located 716:722 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 3 27 theme Asn-X-Thr/Ser 424:436 arg1 sequence 412:419 the signal sequence 401:419 the signal sequence of Asn-X-Thr/Ser 401:436 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 3 28 theme attachment 355:364 arg1 attachment 355:364 attachment 355:364 attachment of glucosamine oligosaccharides 355:396 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 3 28 theme attachment 355:364 arg1 sites 346:350 five sites 341:350 five sites of attachment of glucosamine oligosaccharides 341:396 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 6 29 theme histidine-rich 971:984 arg1 sequence 986:993 a histidine-rich sequence 969:993 a histidine-rich sequence 969:993 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 3 30 theme signal 405:410 arg1 sequence 412:419 the signal sequence 401:419 the signal sequence of Asn-X-Thr/Ser 401:436 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 7 31 contain contains 1182:1189 arg1 regions 1159:1165 four regions 1154:1165 four regions 1154:1165 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 7 31 contain contains 1182:1189 arg2 residues 1216:1223 three or four tryptophan residues 1191:1223 three or four tryptophan residues separated by 0-12 other residues 1191:1256 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 7 31 contain contains 1182:1189 arg1 each 1168:1171 each 1168:1171 each 1168:1171 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 9 32 theme unique 1580:1585 arg1 gene 1598:1601 a unique primordial gene 1578:1601 a unique primordial gene differing from those of other heme-binding proteins 1578:1653 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 9 33 theme significant 1410:1420 arg1 matches 1422:1428 significant matches 1410:1428 significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins 1410:1653 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 0 34 theme hemopexin 19:27 arg1 Structure 0:8 Structure 0:8 Structure of human hemopexin 0:27 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 1 35 theme protein 266:272 arg1 digests 251:257 chemical and enzymatic digests 228:257 chemical and enzymatic digests of the protein 228:272 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 6 36 from sequence 986:993 arg1 present 958:964 present 958:964 present 958:964 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 37 located present 958:964 arg2 oligosaccharides 937:952 the five glucosamine oligosaccharides 916:952 the five glucosamine oligosaccharides 916:952 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 37 located present 958:964 arg1 sequence 986:993 a histidine-rich sequence 969:993 a histidine-rich sequence 969:993 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 37 located present 958:964 arg2 Two 909:911 Two 909:911 Two 909:911 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 5 38 theme middle 807:812 arg1 region 814:819 the middle region 803:819 the middle region 803:819 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 0 39 theme human 13:17 arg1 hemopexin 19:27 human hemopexin 13:27 human hemopexin 13:27 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 0 40 dep sites 56:60 arg1 Structure 0:8 Structure 0:8 Structure of human hemopexin 0:27 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 9 41 theme hemopexin 1439:1447 arg1 matches 1422:1428 significant matches 1410:1428 significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins 1410:1653 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 5 42 theme protein 900:906 arg1 region 886:891 the carboxyl-terminal region 864:891 the carboxyl-terminal region of the protein 864:906 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 0 43 theme O-glycosyl 30:39 arg1 sites 56:60 O-glycosyl and N-glycosyl sites 30:60 O-glycosyl and N-glycosyl sites 30:60 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 0 43 theme O-glycosyl 30:39 arg1 residues 99:106 tryptophan residues 88:106 tryptophan residues 88:106 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 2 44 theme acid 319:322 arg1 residues 324:331 about 440 amino acid residues 303:331 about 440 amino acid residues 303:331 Human hemopexin consists of about 440 amino acid residues. 6371807 5 45 theme amino-terminal 731:744 arg1 region 746:751 the amino-terminal region 727:751 the amino-terminal region 727:751 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 1 46 theme series 195:200 arg1 analysis 181:188 sequence analysis 172:188 sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein 172:272 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 4 47 theme residue 523:529 arg1 feature 459:465 A unique structural feature 439:465 A unique structural feature 439:465 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 47 theme residue 523:529 arg1 blocking 482:489 the virtual blocking 470:489 the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein 470:639 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 48 theme galactosamine 555:567 arg1 oligosaccharide 569:583 a galactosamine oligosaccharide 553:583 a galactosamine oligosaccharide that has not previously been identified in this protein 553:639 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 49 theme structural 448:457 arg1 feature 459:465 A unique structural feature 439:465 A unique structural feature 439:465 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 49 theme structural 448:457 arg1 blocking 482:489 the virtual blocking 470:489 the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein 470:639 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 9 50 theme other 1491:1495 arg1 proteins 1510:1517 other heme-binding proteins 1491:1517 other heme-binding proteins 1491:1517 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 0 51 theme N-glycosyl 45:54 arg1 sites 56:60 O-glycosyl and N-glycosyl sites 30:60 O-glycosyl and N-glycosyl sites 30:60 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 0 51 theme N-glycosyl 45:54 arg1 residues 99:106 tryptophan residues 88:106 tryptophan residues 88:106 Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. 6371807 4 52 theme threonine 513:521 arg1 O-linked 541:548 O-linked 541:548 O-linked 541:548 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 52 theme threonine 513:521 arg1 residue 523:529 the amino-terminal threonine residue 494:529 the amino-terminal threonine residue 494:529 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 6 53 from present 958:964 arg1 sequence 986:993 a histidine-rich sequence 969:993 a histidine-rich sequence 969:993 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 4 54 theme unique 441:446 arg1 feature 459:465 A unique structural feature 439:465 A unique structural feature 439:465 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 4 54 theme unique 441:446 arg1 blocking 482:489 the virtual blocking 470:489 the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein 470:639 A unique structural feature is the virtual blocking of the amino-terminal threonine residue, which is O-linked to a galactosamine oligosaccharide that has not previously been identified in this protein. 6371807 9 55 theme human 1544:1548 arg1 gene 1560:1563 the human hemopexin gene 1540:1563 the human hemopexin gene 1540:1563 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 9 56 theme heme-binding 1497:1508 arg1 proteins 1510:1517 other heme-binding proteins 1491:1517 other heme-binding proteins 1491:1517 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 3 57 theme oligosaccharides 381:396 arg1 attachment 355:364 attachment 355:364 attachment of glucosamine oligosaccharides 355:396 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 9 58 theme computer 1375:1382 arg1 analysis 1384:1391 A computer analysis 1373:1391 A computer analysis 1373:1391 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 5 59 theme glucosamine 684:694 arg1 oligosaccharide 696:710 one glucosamine oligosaccharide 680:710 one glucosamine oligosaccharide 680:710 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 9 60 theme hemopexin 1550:1558 arg1 gene 1560:1563 the human hemopexin gene 1540:1563 the human hemopexin gene 1540:1563 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 6 61 attach present 958:964 arg2 oligosaccharides 937:952 the five glucosamine oligosaccharides 916:952 the five glucosamine oligosaccharides 916:952 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 61 attach present 958:964 arg1 sequence 986:993 a histidine-rich sequence 969:993 a histidine-rich sequence 969:993 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 61 attach present 958:964 arg2 Two 909:911 Two 909:911 Two 909:911 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 1 62 theme primary 113:119 arg1 structure 121:129 The primary structure 109:129 The primary structure of human hemopexin 109:148 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 9 63 theme other 1627:1631 arg1 proteins 1646:1653 other heme-binding proteins 1627:1653 other heme-binding proteins 1627:1653 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 3 64 contain has 337:339 arg1 It 334:335 It 334:335 It 334:335 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 3 64 contain has 337:339 arg2 attachment 355:364 attachment 355:364 attachment of glucosamine oligosaccharides 355:396 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 3 64 contain has 337:339 arg2 sites 346:350 five sites 341:350 five sites of attachment of glucosamine oligosaccharides 341:396 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 2 65 theme amino 313:317 arg1 residues 324:331 about 440 amino acid residues 303:331 about 440 amino acid residues 303:331 Human hemopexin consists of about 440 amino acid residues. 6371807 9 66 theme primordial 1587:1596 arg1 gene 1598:1601 a unique primordial gene 1578:1601 a unique primordial gene differing from those of other heme-binding proteins 1578:1653 A computer analysis did not identify significant matches of human hemopexin to any protein, including cytochromes and other heme-binding proteins, which suggests that the human hemopexin gene evolved from a unique primordial gene differing from those of other heme-binding proteins. 6371807 1 67 theme peptides 205:212 arg1 series 195:200 a series 193:200 a series of peptides obtained from chemical and enzymatic digests of the protein 193:272 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 6 68 theme region 1009:1014 arg1 sequence 986:993 a histidine-rich sequence 969:993 a histidine-rich sequence 969:993 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 1 69 theme human 134:138 arg1 hemopexin 140:148 human hemopexin 134:148 human hemopexin 134:148 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 5 70 theme glucosamine 767:777 arg1 oligosaccharides 779:794 the glucosamine oligosaccharides 763:794 the glucosamine oligosaccharides 763:794 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 3 71 theme glucosamine 369:379 arg1 oligosaccharides 381:396 glucosamine oligosaccharides 369:396 glucosamine oligosaccharides 369:396 It has five sites of attachment of glucosamine oligosaccharides at the signal sequence of Asn-X-Thr/Ser. 6371807 6 72 theme oligosaccharides 937:952 arg1 Two 909:911 Two 909:911 Two 909:911 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 6 72 theme oligosaccharides 937:952 arg1 oligosaccharides 937:952 the five glucosamine oligosaccharides 916:952 the five glucosamine oligosaccharides 916:952 Two of the five glucosamine oligosaccharides are present in a histidine-rich sequence of the middle region of the protein, in which the histidines flank beta-turns presumably at the surface of hemopexin. 6371807 5 73 theme glucosamine 830:840 arg1 oligosaccharide 842:856 one glucosamine oligosaccharide 826:856 one glucosamine oligosaccharide 826:856 The galactosamine oligosaccharide and one glucosamine oligosaccharide are located in the amino-terminal region, three of the glucosamine oligosaccharides are in the middle region, and one glucosamine oligosaccharide is in the carboxyl-terminal region of the protein. 6371807 1 74 theme hemopexin 140:148 arg1 structure 121:129 The primary structure 109:129 The primary structure of human hemopexin 109:148 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 6371807 7 75 theme other 1243:1247 arg1 residues 1249:1256 0-12 other residues 1238:1256 0-12 other residues 1238:1256 Clusters of tryptophan residues occur in four regions, each of which contains three or four tryptophan residues separated by 0-12 other residues. 6371807 1 76 theme chemical 228:235 arg1 digests 251:257 chemical and enzymatic digests 228:257 chemical and enzymatic digests of the protein 228:272 The primary structure of human hemopexin is being deduced from sequence analysis of a series of peptides obtained from chemical and enzymatic digests of the protein. 35377815 0 0 theme von 77:79 arg1 factor 92:97 von Willebrand factor 77:97 von Willebrand factor tubules 77:105 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 5 1 theme domain 733:738 arg1 organization 740:751 shared domain organization 726:751 shared domain organization 726:751 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 10 2 theme assembly 1566:1573 arg1 form 1575:1578 a previously observed supramolecular assembly form 1529:1578 a previously observed supramolecular assembly form 1529:1578 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 2 theme assembly 1566:1573 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 6 3 theme mucin 1032:1036 arg1 region 1022:1027 the amino-terminal region 1003:1027 the amino-terminal region of mucin 1003:1036 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 6 4 theme VWF 952:954 arg1 adaptation 956:965 a VWF adaptation 950:965 a VWF adaptation to the vasculature 950:984 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 7 5 theme human 1059:1063 arg1 MUC2 1095:1098 the human intestinal gel-forming mucin (MUC2) 1055:1099 the human intestinal gel-forming mucin (MUC2) 1055:1099 This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. 35377815 3 6 from sites 471:475 arg1 collagen 453:460 subendothelial collagen 438:460 subendothelial collagen at wound sites 438:475 When VWF is released into the bloodstream, these tubules unfurl to release linear polymers that bind subendothelial collagen at wound sites, recruit platelets, and initiate the clotting cascade. 35377815 8 7 theme residue-resolution 1239:1256 arg1 structure 1258:1266 the residue-resolution structure 1235:1266 the residue-resolution structure of tubules formed by the homologous segment of VWF 1235:1317 To facilitate a comparison, we determined the residue-resolution structure of tubules formed by the homologous segment of VWF. 35377815 2 8 from tubules 288:294 arg1 cells 311:315 endothelial cells 299:315 endothelial cells prior to secretion 299:334 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 5 9 theme VWF 672:674 arg1 function 660:667 the divergent function 646:667 the divergent function of VWF in blood vessel repair 646:697 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 0 10 theme factor 92:97 arg1 tubules 99:105 von Willebrand factor tubules 77:105 von Willebrand factor tubules 77:105 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 9 11 theme tubules 1355:1361 arg1 structures 1324:1333 The structures 1320:1333 The structures of the MUC2 and VWF tubules 1320:1361 The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. 35377815 8 12 theme tubules 1271:1277 arg1 structure 1258:1266 the residue-resolution structure 1235:1266 the residue-resolution structure of tubules formed by the homologous segment of VWF 1235:1317 To facilitate a comparison, we determined the residue-resolution structure of tubules formed by the homologous segment of VWF. 35377815 9 13 theme MUC2 1342:1345 arg1 tubules 1355:1361 the MUC2 and VWF tubules 1338:1361 the MUC2 and VWF tubules 1338:1361 The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. 35377815 0 14 theme Willebrand 81:90 arg1 factor 92:97 von Willebrand factor 77:97 von Willebrand factor tubules 77:105 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 11 15 from mucins 1788:1793 arg1 present 1777:1783 present 1777:1783 present 1777:1783 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 7 16 theme mucin 1088:1092 arg1 MUC2 1095:1098 the human intestinal gel-forming mucin (MUC2) 1055:1099 the human intestinal gel-forming mucin (MUC2) 1055:1099 This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. 35377815 4 17 theme gel-forming 549:559 arg1 mucins 561:566 gel-forming mucins 549:566 gel-forming mucins 549:566 VWF evolved from gel-forming mucins, the polymeric glycoproteins that coat and protect exposed epithelia. 35377815 5 18 theme sequence 700:707 arg1 conservation 709:720 sequence conservation 700:720 sequence conservation 700:720 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 9 19 theme intermolecular 1400:1413 arg1 interactions 1415:1426 the intermolecular interactions 1396:1426 the intermolecular interactions required for tubule formation 1396:1456 The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. 35377815 4 20 gly glycoproteins 583:595 arg1 glycoproteins 583:595 the polymeric glycoproteins 569:595 the polymeric glycoproteins that coat and protect exposed epithelia 569:635 VWF evolved from gel-forming mucins, the polymeric glycoproteins that coat and protect exposed epithelia. 35377815 10 21 theme full-length 1481:1491 arg1 MUC2 1493:1496 full-length MUC2 1481:1496 full-length MUC2 1481:1496 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 6 22 theme region 1022:1027 arg1 ability 879:885 the ability 875:885 the ability to form tubules 875:901 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 6 22 theme region 1022:1027 arg1 feature 992:998 a feature 990:998 a feature of the amino-terminal region of mucin 990:1036 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 10 23 theme supramolecular 1551:1564 arg1 form 1575:1578 a previously observed supramolecular assembly form 1529:1578 a previously observed supramolecular assembly form 1529:1578 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 23 theme supramolecular 1551:1564 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 6 24 theme cryo-electron 845:857 arg1 microscopy 859:868 cryo-electron microscopy 845:868 cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin 845:1036 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 6 25 dep ability 879:885 arg1 form 890:893 form 890:893 to form tubules 887:901 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 10 26 theme physiological 1620:1632 arg1 intermediate 1648:1659 the physiological mucin storage intermediate 1616:1659 the physiological mucin storage intermediate 1616:1659 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 26 theme physiological 1620:1632 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 11 27 theme VWF 1733:1735 arg1 tubules 1737:1743 VWF tubules 1733:1743 VWF tubules 1733:1743 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 4 28 theme exposed 619:625 arg1 epithelia 627:635 exposed epithelia 619:635 exposed epithelia 619:635 VWF evolved from gel-forming mucins, the polymeric glycoproteins that coat and protect exposed epithelia. 35377815 5 29 theme key 777:779 arg1 aspects 781:787 key aspects 777:787 key aspects of the mucin bioassembly mechanism 777:822 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 10 30 theme linear 1511:1516 arg1 intermediate 1648:1659 the physiological mucin storage intermediate 1616:1659 the physiological mucin storage intermediate 1616:1659 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 30 theme linear 1511:1516 arg1 form 1575:1578 a previously observed supramolecular assembly form 1529:1578 a previously observed supramolecular assembly form 1529:1578 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 30 theme linear 1511:1516 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 11 31 from VWF 1799:1801 arg1 present 1777:1783 present 1777:1783 present 1777:1783 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 8 32 theme homologous 1293:1302 arg1 segment 1304:1310 the homologous segment 1289:1310 the homologous segment of VWF 1289:1317 To facilitate a comparison, we determined the residue-resolution structure of tubules formed by the homologous segment of VWF. 35377815 5 33 theme divergent 650:658 arg1 function 660:667 the divergent function 646:667 the divergent function of VWF in blood vessel repair 646:697 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 0 34 theme Helical 0:6 arg1 self-assembly 8:20 Helical self-assembly 0:20 Helical self-assembly of a mucin segment 0:39 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 10 35 from constraints 1466:1476 arg1 MUC2 1493:1496 full-length MUC2 1481:1496 full-length MUC2 1481:1496 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 7 36 theme MUC2 1095:1098 arg1 segment 1044:1050 This segment 1039:1050 This segment of the human intestinal gel-forming mucin (MUC2) 1039:1099 This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. 35377815 11 37 theme design 1759:1764 arg1 principles 1766:1775 design principles 1759:1775 design principles present in mucins and VWF 1759:1801 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 1 38 gly glycoprotein 112:123 arg1 glycoprotein 112:123 The glycoprotein von Willebrand factor 108:145 The glycoprotein von Willebrand factor (VWF) 108:151 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 9 39 theme VWF 1351:1353 arg1 tubules 1355:1361 the MUC2 and VWF tubules 1338:1361 the MUC2 and VWF tubules 1338:1361 The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. 35377815 3 40 theme clotting 514:521 arg1 cascade 523:529 the clotting cascade 510:529 the clotting cascade 510:529 When VWF is released into the bloodstream, these tubules unfurl to release linear polymers that bind subendothelial collagen at wound sites, recruit platelets, and initiate the clotting cascade. 35377815 5 41 theme mucin 796:800 arg1 mechanism 814:822 the mucin bioassembly mechanism 792:822 the mucin bioassembly mechanism 792:822 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 11 42 theme possible 1700:1707 arg1 origin 1722:1727 a possible evolutionary origin 1698:1727 a possible evolutionary origin for VWF tubules 1698:1743 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 11 43 theme evolutionary 1709:1720 arg1 origin 1722:1727 a possible evolutionary origin 1698:1727 a possible evolutionary origin for VWF tubules 1698:1743 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 2 44 theme long 274:277 arg1 tubules 288:294 long, helical tubules 274:294 long, helical tubules in endothelial cells prior to secretion 274:334 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 5 45 theme bioassembly 802:812 arg1 mechanism 814:822 the mucin bioassembly mechanism 792:822 the mucin bioassembly mechanism 792:822 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 0 46 theme segment 33:39 arg1 self-assembly 8:20 Helical self-assembly 0:20 Helical self-assembly of a mucin segment 0:39 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 2 47 dep long 274:277 arg1 helical 280:286 helical 280:286 helical 280:286 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 5 48 theme blood 679:683 arg1 repair 692:697 blood vessel repair 679:697 blood vessel repair 679:697 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 5 49 theme mechanism 814:822 arg1 aspects 781:787 key aspects 777:787 key aspects of the mucin bioassembly mechanism 777:822 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 3 50 theme linear 412:417 arg1 polymers 419:426 linear polymers 412:426 linear polymers that bind subendothelial collagen at wound sites 412:475 When VWF is released into the bloodstream, these tubules unfurl to release linear polymers that bind subendothelial collagen at wound sites, recruit platelets, and initiate the clotting cascade. 35377815 0 51 theme mucin 27:31 arg1 segment 33:39 a mucin segment 25:39 a mucin segment 25:39 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 11 52 attach present 1777:1783 arg1 VWF 1799:1801 VWF 1799:1801 VWF 1799:1801 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 11 52 attach present 1777:1783 arg2 principles 1766:1775 design principles 1759:1775 design principles present in mucins and VWF 1759:1801 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 11 52 attach present 1777:1783 arg1 mucins 1788:1793 mucins 1788:1793 mucins 1788:1793 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 5 53 theme vessel 685:690 arg1 repair 692:697 blood vessel repair 679:697 blood vessel repair 679:697 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 8 54 theme VWF 1315:1317 arg1 segment 1304:1310 the homologous segment 1289:1310 the homologous segment of VWF 1289:1317 To facilitate a comparison, we determined the residue-resolution structure of tubules formed by the homologous segment of VWF. 35377815 1 55 theme blood 204:208 arg1 walls 217:221 blood vessel walls 204:221 blood vessel walls 204:221 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 7 56 theme intestinal 1065:1074 arg1 MUC2 1095:1098 the human intestinal gel-forming mucin (MUC2) 1055:1099 the human intestinal gel-forming mucin (MUC2) 1055:1099 This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. 35377815 6 57 dep microscopy 859:868 arg1 ability 879:885 the ability 875:885 the ability to form tubules 875:901 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 6 57 dep microscopy 859:868 arg1 feature 992:998 a feature 990:998 a feature of the amino-terminal region of mucin 990:1036 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 10 58 theme Steric 1459:1464 arg1 constraints 1466:1476 Steric constraints 1459:1476 Steric constraints in full-length MUC2 1459:1496 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 5 59 theme shared 726:731 arg1 organization 740:751 shared domain organization 726:751 shared domain organization 726:751 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 10 60 theme observed 1542:1549 arg1 form 1575:1578 a previously observed supramolecular assembly form 1529:1578 a previously observed supramolecular assembly form 1529:1578 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 60 theme observed 1542:1549 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 2 61 theme prior 317:321 arg1 cells 311:315 endothelial cells 299:315 endothelial cells prior to secretion 299:334 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 7 62 theme gel-forming 1076:1086 arg1 MUC2 1095:1098 the human intestinal gel-forming mucin (MUC2) 1055:1099 the human intestinal gel-forming mucin (MUC2) 1055:1099 This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. 35377815 3 63 theme subendothelial 438:451 arg1 collagen 453:460 subendothelial collagen 438:460 subendothelial collagen at wound sites 438:475 When VWF is released into the bloodstream, these tubules unfurl to release linear polymers that bind subendothelial collagen at wound sites, recruit platelets, and initiate the clotting cascade. 35377815 1 64 theme glycoprotein 112:123 arg1 VWF 148:150 VWF 148:150 VWF 148:150 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 1 64 theme glycoprotein 112:123 arg1 factor 140:145 The glycoprotein von Willebrand factor 108:145 The glycoprotein von Willebrand factor (VWF) 108:151 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 11 65 theme present 1777:1783 arg1 principles 1766:1775 design principles 1759:1775 design principles present in mucins and VWF 1759:1801 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 11 66 theme MUC2 1676:1679 arg1 tubules 1681:1687 MUC2 tubules 1676:1687 MUC2 tubules 1676:1687 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 2 67 theme VWF 249:251 arg1 assembly 260:267 its assembly 256:267 its assembly into long, helical tubules in endothelial cells prior to secretion 256:334 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 2 67 theme VWF 249:251 arg1 feature 238:244 A distinctive feature 224:244 A distinctive feature of VWF 224:251 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 1 68 from injuries 192:199 arg1 walls 217:221 blood vessel walls 204:221 blood vessel walls 204:221 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 1 69 theme von 125:127 arg1 VWF 148:150 VWF 148:150 VWF 148:150 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 1 69 theme von 125:127 arg1 factor 140:145 The glycoprotein von Willebrand factor 108:145 The glycoprotein von Willebrand factor (VWF) 108:151 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 1 70 theme vessel 210:215 arg1 walls 217:221 blood vessel walls 204:221 blood vessel walls 204:221 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 11 71 from present 1777:1783 arg1 VWF 1799:1801 VWF 1799:1801 VWF 1799:1801 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 11 71 from present 1777:1783 arg1 mucins 1788:1793 mucins 1788:1793 mucins 1788:1793 Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF. 35377815 1 72 theme Willebrand 129:138 arg1 VWF 148:150 VWF 148:150 VWF 148:150 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 1 72 theme Willebrand 129:138 arg1 factor 140:145 The glycoprotein von Willebrand factor 108:145 The glycoprotein von Willebrand factor (VWF) 108:151 The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. 35377815 2 73 theme endothelial 299:309 arg1 cells 311:315 endothelial cells 299:315 endothelial cells prior to secretion 299:334 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 7 74 theme striking 1148:1155 arg1 resemblance 1157:1167 a striking resemblance 1146:1167 a striking resemblance to those of VWF itself 1146:1190 This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. 35377815 10 75 theme storage 1640:1646 arg1 intermediate 1648:1659 the physiological mucin storage intermediate 1616:1659 the physiological mucin storage intermediate 1616:1659 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 75 theme storage 1640:1646 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 9 76 theme flexible 1376:1383 arg1 joints 1385:1390 the flexible joints 1372:1390 the flexible joints 1372:1390 The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. 35377815 9 77 theme tubule 1441:1446 arg1 formation 1448:1456 tubule formation 1441:1456 tubule formation 1441:1456 The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. 35377815 6 78 theme amino-terminal 1007:1020 arg1 region 1022:1027 the amino-terminal region 1003:1027 the amino-terminal region of mucin 1003:1036 Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. 35377815 0 79 theme evolutionary 53:64 arg1 origin 66:71 an evolutionary origin 50:71 an evolutionary origin for von Willebrand factor tubules 50:105 Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules. 35377815 10 80 theme mucin 1634:1638 arg1 intermediate 1648:1659 the physiological mucin storage intermediate 1616:1659 the physiological mucin storage intermediate 1616:1659 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 10 80 theme mucin 1634:1638 arg1 filaments 1518:1526 linear filaments 1511:1526 linear filaments 1511:1526 Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. 35377815 2 81 theme distinctive 226:236 arg1 assembly 260:267 its assembly 256:267 its assembly into long, helical tubules in endothelial cells prior to secretion 256:334 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 2 81 theme distinctive 226:236 arg1 feature 238:244 A distinctive feature 224:244 A distinctive feature of VWF 224:251 A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. 35377815 5 82 from function 660:667 arg1 repair 692:697 blood vessel repair 679:697 blood vessel repair 679:697 Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. 35377815 3 83 theme wound 465:469 arg1 sites 471:475 wound sites 465:475 wound sites 465:475 When VWF is released into the bloodstream, these tubules unfurl to release linear polymers that bind subendothelial collagen at wound sites, recruit platelets, and initiate the clotting cascade. 35377815 4 84 theme polymeric 573:581 arg1 glycoproteins 583:595 the polymeric glycoproteins 569:595 the polymeric glycoproteins that coat and protect exposed epithelia 569:635 VWF evolved from gel-forming mucins, the polymeric glycoproteins that coat and protect exposed epithelia. 25727495 0 0 theme acyltransferase 78:92 arg1 function 14:21 function 14:21 function 14:21 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 0 0 theme acyltransferase 78:92 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 5 1 theme human 932:936 arg1 disease 938:944 human disease 932:944 human disease 932:944 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 4 2 theme LPLA2 736:740 arg1 core 728:731 the α/β hydrolase core 710:731 the α/β hydrolase core of LPLA2 710:740 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 1 3 theme cholesterol 143:153 arg1 LCAT 172:175 LCAT 172:175 LCAT 172:175 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 3 theme cholesterol 143:153 arg1 acyltransferase 155:169 lecithin:cholesterol acyltransferase 134:169 lecithin:cholesterol acyltransferase (LCAT) 134:176 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 4 theme enzymes 252:258 arg1 family 219:224 a structurally uncharacterized family 188:224 a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport 188:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 0 5 theme cholesterol 66:76 arg1 acyltransferase 78:92 lecithin:cholesterol acyltransferase 57:92 lecithin:cholesterol acyltransferase 57:92 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 1 6 theme responsible 260:270 arg1 enzymes 252:258 key lipid-metabolizing enzymes 229:258 key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport 229:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 5 7 theme therapeutics 1043:1054 arg1 development 1024:1034 rational development 1015:1034 rational development of new therapeutics 1015:1054 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 3 8 theme several 541:547 arg1 structures 573:582 several high-resolution crystal structures 541:582 several high-resolution crystal structures of human LPLA2 541:597 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 3 9 theme human 587:591 arg1 LPLA2 593:597 human LPLA2 587:597 human LPLA2 587:597 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 1 10 theme lung 276:279 arg1 catabolism 292:301 lung surfactant catabolism 276:301 lung surfactant catabolism 276:301 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 5 11 theme LCAT 877:880 arg1 structure 882:890 The LCAT structure 873:890 The LCAT structure 873:890 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 3 12 theme crystal 565:571 arg1 structures 573:582 several high-resolution crystal structures 541:582 several high-resolution crystal structures of human LPLA2 541:597 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 2 13 theme drug-induced 414:425 arg1 phospholipidosis 427:442 drug-induced phospholipidosis 414:442 drug-induced phospholipidosis 414:442 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 2 14 theme eye 482:484 arg1 disease 486:492 fish eye disease 477:492 fish eye disease 477:492 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 2 15 theme familial 498:505 arg1 deficiency 512:521 familial LCAT deficiency 498:521 familial LCAT deficiency 498:521 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 5 16 theme molecular 905:913 arg1 basis 915:919 the molecular basis 901:919 the molecular basis underlying human disease for most of the known LCAT missense mutations 901:990 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 2 17 theme fish 477:480 arg1 disease 486:492 fish eye disease 477:492 fish eye disease 477:492 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 2 18 theme LCAT 507:510 arg1 deficiency 512:521 familial LCAT deficiency 498:521 familial LCAT deficiency 498:521 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 5 19 theme missense 973:980 arg1 mutations 982:990 the known LCAT missense mutations 958:990 the known LCAT missense mutations 958:990 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 4 20 from Insertions 696:705 arg1 core 728:731 the α/β hydrolase core 710:731 the α/β hydrolase core of LPLA2 710:740 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 21 theme α/β 714:716 arg1 core 728:731 the α/β hydrolase core 710:731 the α/β hydrolase core of LPLA2 710:740 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 1 22 theme surfactant 281:290 arg1 catabolism 292:301 lung surfactant catabolism 276:301 lung surfactant catabolism 276:301 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 23 theme Lysosomal 95:103 arg1 LPLA2 123:127 LPLA2 123:127 LPLA2 123:127 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 23 theme Lysosomal 95:103 arg1 A2 119:120 Lysosomal phospholipase A2 95:120 Lysosomal phospholipase A2 (LPLA2) 95:128 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 4 24 theme responsible 764:774 arg1 domains 747:753 domains 747:753 domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates 747:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 24 theme responsible 764:774 arg1 chains 822:827 the acyl chains 813:827 the acyl chains 813:827 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 5 25 theme LCAT 968:971 arg1 mutations 982:990 the known LCAT missense mutations 958:990 the known LCAT missense mutations 958:990 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 1 26 theme phospholipase 105:117 arg1 LPLA2 123:127 LPLA2 123:127 LPLA2 123:127 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 26 theme phospholipase 105:117 arg1 A2 119:120 Lysosomal phospholipase A2 95:120 Lysosomal phospholipase A2 (LPLA2) 95:128 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 4 27 theme phospholipid 848:859 arg1 substrates 861:870 phospholipid substrates 848:870 phospholipid substrates 848:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 3 28 theme low-resolution 605:618 arg1 structure 620:628 a low-resolution structure 603:628 a low-resolution structure of LCAT 603:636 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 4 29 theme acyl 817:820 arg1 domains 747:753 domains 747:753 domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates 747:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 29 theme acyl 817:820 arg1 chains 822:827 the acyl chains 813:827 the acyl chains 813:827 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 5 30 theme acute 1102:1106 arg1 syndrome 1117:1124 acute coronary syndrome 1102:1124 acute coronary syndrome 1102:1124 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 4 31 theme membrane 780:787 arg1 interaction 789:799 membrane interaction 780:799 membrane interaction 780:799 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 2 32 theme somatic 445:451 arg1 mutations 453:461 somatic mutations 445:461 somatic mutations in LCAT 445:469 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 0 33 theme phospholipase 36:48 arg1 A2 50:51 lysosomal phospholipase A2 26:51 lysosomal phospholipase A2 26:51 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 5 34 theme coronary 1108:1115 arg1 syndrome 1117:1124 acute coronary syndrome 1102:1124 acute coronary syndrome 1102:1124 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 3 35 theme LCAT 633:636 arg1 structure 620:628 a low-resolution structure 603:628 a low-resolution structure of LCAT 603:636 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 3 35 theme LCAT 633:636 arg1 structures 573:582 several high-resolution crystal structures 541:582 several high-resolution crystal structures of human LPLA2 541:597 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 0 36 theme lysosomal 26:34 arg1 A2 50:51 lysosomal phospholipase A2 26:51 lysosomal phospholipase A2 26:51 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 1 37 theme reverse 311:317 arg1 transport 331:339 reverse cholesterol transport 311:339 reverse cholesterol transport 311:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 38 theme uncharacterized 203:217 arg1 family 219:224 a structurally uncharacterized family 188:224 a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport 188:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 39 theme cholesterol 319:329 arg1 transport 331:339 reverse cholesterol transport 311:339 reverse cholesterol transport 311:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 0 40 theme A2 50:51 arg1 function 14:21 function 14:21 function 14:21 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 0 40 theme A2 50:51 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 5 41 theme LCAT 1065:1068 arg1 deficiency 1070:1079 LCAT deficiency 1065:1079 LCAT deficiency 1065:1079 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 4 42 theme substrates 861:870 arg1 domains 747:753 domains 747:753 domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates 747:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 42 theme substrates 861:870 arg1 chains 822:827 the acyl chains 813:827 the acyl chains 813:827 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 42 theme substrates 861:870 arg1 groups 838:843 head groups 833:843 head groups of phospholipid substrates 833:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 42 theme substrates 861:870 arg1 substrates 861:870 phospholipid substrates 848:870 phospholipid substrates 848:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 5 43 theme new 1039:1041 arg1 therapeutics 1043:1054 new therapeutics 1039:1054 new therapeutics 1039:1054 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 5 44 theme rational 1015:1022 arg1 development 1024:1034 rational development 1015:1034 rational development of new therapeutics 1015:1054 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 3 45 theme close 656:660 arg1 relationship 673:684 its close structural relationship 652:684 its close structural relationship to LPLA2 652:693 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 5 46 theme known 962:966 arg1 mutations 982:990 the known LCAT missense mutations 958:990 the known LCAT missense mutations 958:990 The LCAT structure suggests the molecular basis underlying human disease for most of the known LCAT missense mutations, and paves the way for rational development of new therapeutics to treat LCAT deficiency, atherosclerosis and acute coronary syndrome. 25727495 3 47 theme LPLA2 593:597 arg1 structure 620:628 a low-resolution structure 603:628 a low-resolution structure of LCAT 603:636 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 3 47 theme LPLA2 593:597 arg1 structures 573:582 several high-resolution crystal structures 541:582 several high-resolution crystal structures of human LPLA2 541:597 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 3 48 theme structural 662:671 arg1 relationship 673:684 its close structural relationship 652:684 its close structural relationship to LPLA2 652:693 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 4 49 theme hydrolase 718:726 arg1 core 728:731 the α/β hydrolase core 710:731 the α/β hydrolase core of LPLA2 710:740 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 2 50 theme phospholipidosis 427:442 arg1 development 399:409 the development 395:409 the development of drug-induced phospholipidosis 395:442 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 1 51 theme lecithin 134:141 arg1 LCAT 172:175 LCAT 172:175 LCAT 172:175 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 51 theme lecithin 134:141 arg1 acyltransferase 155:169 lecithin:cholesterol acyltransferase 134:169 lecithin:cholesterol acyltransferase (LCAT) 134:176 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 1 52 theme key 229:231 arg1 enzymes 252:258 key lipid-metabolizing enzymes 229:258 key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport 229:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 25727495 0 53 theme lecithin 57:64 arg1 acyltransferase 78:92 lecithin:cholesterol acyltransferase 57:92 lecithin:cholesterol acyltransferase 57:92 Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase. 25727495 4 54 theme head 833:836 arg1 groups 838:843 head groups 833:843 head groups of phospholipid substrates 833:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 4 54 theme head 833:836 arg1 substrates 861:870 phospholipid substrates 848:870 phospholipid substrates 848:870 Insertions in the α/β hydrolase core of LPLA2 form domains that are responsible for membrane interaction and binding the acyl chains and head groups of phospholipid substrates. 25727495 2 55 from mutations 453:461 arg1 LCAT 466:469 LCAT 466:469 LCAT 466:469 Whereas LPLA2 is predicted to underlie the development of drug-induced phospholipidosis, somatic mutations in LCAT cause fish eye disease and familial LCAT deficiency. 25727495 3 56 theme high-resolution 549:563 arg1 structures 573:582 several high-resolution crystal structures 541:582 several high-resolution crystal structures of human LPLA2 541:597 Here we describe several high-resolution crystal structures of human LPLA2 and a low-resolution structure of LCAT that confirms its close structural relationship to LPLA2. 25727495 1 57 theme lipid-metabolizing 233:250 arg1 enzymes 252:258 key lipid-metabolizing enzymes 229:258 key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport 229:339 Lysosomal phospholipase A2 (LPLA2) and lecithin:cholesterol acyltransferase (LCAT) belong to a structurally uncharacterized family of key lipid-metabolizing enzymes responsible for lung surfactant catabolism and for reverse cholesterol transport, respectively. 8962717 0 0 theme epidermal 74:82 arg1 receptor 98:105 the epidermal growth factor receptor 70:105 the epidermal growth factor receptor 70:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 4 1 theme molecular 671:679 arg1 weight 681:686 the molecular weight 667:686 the molecular weight from 110 kDa 667:699 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 5 2 theme pulsed 823:828 arg1 detection 843:851 pulsed amperometric detection 823:851 pulsed amperometric detection 823:851 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 2 3 theme ovary 345:349 arg1 cells 351:355 Chinese hamster ovary cells 329:355 Chinese hamster ovary cells 329:355 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 1 4 theme factor 238:243 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 5 5 theme high 760:763 arg1 sensitivity 765:775 high sensitivity 760:775 high sensitivity 760:775 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 1 6 theme extracellular 159:171 arg1 acids 202:206 621 N-terminal amino acids 181:206 621 N-terminal amino acids 181:206 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 6 theme extracellular 159:171 arg1 domain 173:178 The extracellular domain 155:178 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor 155:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 6 theme extracellular 159:171 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 0 7 theme factor 91:96 arg1 receptor 98:105 the epidermal growth factor receptor 70:105 the epidermal growth factor receptor 70:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 3 8 theme digests 536:542 arg1 separation 482:491 chromatographic separation 466:491 chromatographic separation 466:491 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 3 8 theme digests 536:542 arg1 analysis 516:523 mass spectrometric analysis 497:523 mass spectrometric analysis 497:523 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 1 9 theme EGF 246:248 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 0 10 theme growth 84:89 arg1 receptor 98:105 the epidermal growth factor receptor 70:105 the epidermal growth factor receptor 70:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 6 11 theme terminal 1032:1039 arg1 GlcNAc 1041:1046 the reducing terminal GlcNAc 1019:1046 the reducing terminal GlcNAc 1019:1046 The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. 8962717 5 12 theme Released 712:719 arg1 oligosaccharides 721:736 Released oligosaccharides 712:736 Released oligosaccharides 712:736 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 3 13 theme chromatographic 466:480 arg1 separation 482:491 chromatographic separation 466:491 chromatographic separation 466:491 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 5 14 theme amperometric 830:841 arg1 detection 843:851 pulsed amperometric detection 823:851 pulsed amperometric detection 823:851 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 4 15 theme peptide-N-glycosidase 623:643 arg1 endoglycosidase 606:620 the endoglycosidase 602:620 the endoglycosidase 602:620 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 4 15 theme peptide-N-glycosidase 623:643 arg1 F 645:645 peptide-N-glycosidase F 623:645 peptide-N-glycosidase F (PNGase F) 623:656 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 4 15 theme peptide-N-glycosidase 623:643 arg1 F 655:655 PNGase F 648:655 PNGase F 648:655 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 1 16 contain has 260:262 arg1 acids 202:206 621 N-terminal amino acids 181:206 621 N-terminal amino acids 181:206 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 16 contain has 260:262 arg1 domain 173:178 The extracellular domain 155:178 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor 155:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 16 contain has 260:262 arg2 sites 304:308 eleven consensus N-linked glycosylation sites 264:308 eleven consensus N-linked glycosylation sites 264:308 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 16 contain has 260:262 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 5 17 theme gas-liquid 857:866 arg1 spectrometry 888:899 gas-liquid chromatography/mass spectrometry 857:899 gas-liquid chromatography/mass spectrometry 857:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 0 18 theme receptor 98:105 arg1 domain 60:65 the extracellular domain 42:65 the extracellular domain of the epidermal growth factor receptor 42:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 0 18 theme receptor 98:105 arg1 receptor 98:105 the epidermal growth factor receptor 70:105 the epidermal growth factor receptor 70:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 3 19 theme tryptic 528:534 arg1 digests 536:542 tryptic digests 528:542 tryptic digests 528:542 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 5 20 theme chromatography/mass 868:886 arg1 spectrometry 888:899 gas-liquid chromatography/mass spectrometry 857:899 gas-liquid chromatography/mass spectrometry 857:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 5 21 theme anion 788:792 arg1 chromatography 803:816 high pH anion exchange chromatography 780:816 high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry 780:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 6 22 theme reducing 1023:1030 arg1 GlcNAc 1041:1046 the reducing terminal GlcNAc 1019:1046 the reducing terminal GlcNAc 1019:1046 The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. 8962717 5 23 with chromatography 803:816 arg1 detection 843:851 pulsed amperometric detection 823:851 pulsed amperometric detection 823:851 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 5 23 with chromatography 803:816 arg1 spectrometry 888:899 gas-liquid chromatography/mass spectrometry 857:899 gas-liquid chromatography/mass spectrometry 857:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 2 24 theme chains 426:431 arg1 combination 386:396 a combination 384:396 a combination of high mannose and complex chains 384:431 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 1 25 theme receptor 251:258 arg1 acids 202:206 621 N-terminal amino acids 181:206 621 N-terminal amino acids 181:206 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 25 theme receptor 251:258 arg1 domain 173:178 The extracellular domain 155:178 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor 155:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 25 theme receptor 251:258 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 2 26 gly glycosylated 366:377 arg1 this 357:360 this 357:360 this 357:360 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 6 27 gly fucosylated 1004:1014 arg1 oligosaccharides 987:1002 tetra-antennary oligosaccharides 971:1002 tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc 971:1046 The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. 8962717 1 28 theme N-terminal 185:194 arg1 acids 202:206 621 N-terminal amino acids 181:206 621 N-terminal amino acids 181:206 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 28 theme N-terminal 185:194 arg1 domain 173:178 The extracellular domain 155:178 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor 155:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 28 theme N-terminal 185:194 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 0 29 theme glycosylation 16:28 arg1 patterns 30:37 the glycosylation patterns 12:37 the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor 12:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 5 30 theme exchange 794:801 arg1 chromatography 803:816 high pH anion exchange chromatography 780:816 high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry 780:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 0 31 theme Chinese 120:126 arg1 fibroblasts 142:152 Chinese hamster ovary fibroblasts 120:152 Chinese hamster ovary fibroblasts 120:152 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 6 32 gly trisialylated 957:969 arg1 chains 944:949 the complex chains 932:949 the complex chains 932:949 The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. 8962717 5 33 theme high 780:783 arg1 chromatography 803:816 high pH anion exchange chromatography 780:816 high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry 780:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 1 34 theme amino 196:200 arg1 acids 202:206 621 N-terminal amino acids 181:206 621 N-terminal amino acids 181:206 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 34 theme amino 196:200 arg1 domain 173:178 The extracellular domain 155:178 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor 155:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 34 theme amino 196:200 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 7 35 theme ligand 1119:1124 arg1 binding 1126:1132 ligand binding 1119:1132 ligand binding 1119:1132 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 35 theme ligand 1119:1124 arg1 effect 1138:1143 an effect 1135:1143 an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4 1135:1292 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 35 theme ligand 1119:1124 arg1 likely 1154:1159 likely 1154:1159 likely 1154:1159 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 36 theme large 1053:1057 arg1 mass 1072:1075 The large hydrodynamic mass 1049:1075 The large hydrodynamic mass of these oligosaccharides 1049:1101 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 2 37 theme hamster 337:343 arg1 cells 351:355 Chinese hamster ovary cells 329:355 Chinese hamster ovary cells 329:355 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 6 38 theme complex 936:942 arg1 chains 944:949 the complex chains 932:949 the complex chains 932:949 The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. 8962717 2 39 theme high 401:404 arg1 mannose 406:412 high mannose 401:412 high mannose 401:412 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 7 40 theme consensus 1192:1200 arg1 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 40 theme consensus 1192:1200 arg1 sites 1216:1220 consensus glycosylation sites 1192:1220 consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4 1192:1292 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 1 41 theme consensus 271:279 arg1 sites 304:308 eleven consensus N-linked glycosylation sites 264:308 eleven consensus N-linked glycosylation sites 264:308 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 0 42 theme ovary 136:140 arg1 fibroblasts 142:152 Chinese hamster ovary fibroblasts 120:152 Chinese hamster ovary fibroblasts 120:152 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 7 43 theme related 1234:1240 arg1 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 44 gly glycosylation 1202:1214 arg2 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 44 gly glycosylation 1202:1214 arg1 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 44 gly glycosylation 1202:1214 arg2 sites 1216:1220 consensus glycosylation sites 1192:1220 consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4 1192:1292 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 4 45 theme PNGase 648:653 arg1 F 645:645 peptide-N-glycosidase F 623:645 peptide-N-glycosidase F (PNGase F) 623:656 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 4 45 theme PNGase 648:653 arg1 F 655:655 PNGase F 648:655 PNGase F 648:655 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 0 46 theme patterns 30:37 arg1 Analysis 0:7 Analysis 0:7 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor 0:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 1 47 theme N-linked 281:288 arg1 sites 304:308 eleven consensus N-linked glycosylation sites 264:308 eleven consensus N-linked glycosylation sites 264:308 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 0 48 theme hamster 128:134 arg1 fibroblasts 142:152 Chinese hamster ovary fibroblasts 120:152 Chinese hamster ovary fibroblasts 120:152 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 2 49 theme complex 418:424 arg1 chains 426:431 complex chains 418:431 complex chains 418:431 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 3 50 theme latter 438:443 arg1 chains 445:450 The latter chains 434:450 The latter chains 434:450 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 0 51 theme extracellular 46:58 arg1 domain 60:65 the extracellular domain 42:65 the extracellular domain of the epidermal growth factor receptor 42:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 0 51 theme extracellular 46:58 arg1 receptor 98:105 the epidermal growth factor receptor 70:105 the epidermal growth factor receptor 70:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 1 52 theme glycosylation 290:302 arg1 sites 304:308 eleven consensus N-linked glycosylation sites 264:308 eleven consensus N-linked glycosylation sites 264:308 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 3 53 theme EGF-binding 567:577 arg1 domain 579:584 the EGF-binding domain 563:584 the EGF-binding domain 563:584 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 2 54 theme Chinese 329:335 arg1 cells 351:355 Chinese hamster ovary cells 329:355 Chinese hamster ovary cells 329:355 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 5 55 theme pH 785:786 arg1 chromatography 803:816 high pH anion exchange chromatography 780:816 high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry 780:899 Released oligosaccharides were characterised at high sensitivity by high pH anion exchange chromatography with pulsed amperometric detection and gas-liquid chromatography/mass spectrometry. 8962717 0 56 gly glycosylation 16:28 arg1 domain 60:65 the extracellular domain 42:65 the extracellular domain of the epidermal growth factor receptor 42:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 0 56 gly glycosylation 16:28 arg1 receptor 98:105 the epidermal growth factor receptor 70:105 the epidermal growth factor receptor 70:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 1 57 link N-linked 281:288 arg1 sites 304:308 eleven consensus N-linked glycosylation sites 264:308 eleven consensus N-linked glycosylation sites 264:308 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 7 58 dep p185erbB2/neu 1255:1267 arg1 i.e. 1250:1253 i.e. 1250:1253 i.e. 1250:1253 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 3 59 theme mass 497:500 arg1 analysis 516:523 mass spectrometric analysis 497:523 mass spectrometric analysis 497:523 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 4 60 with Treatment 587:595 arg1 endoglycosidase 606:620 the endoglycosidase 602:620 the endoglycosidase 602:620 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 4 60 with Treatment 587:595 arg1 F 645:645 peptide-N-glycosidase F 623:645 peptide-N-glycosidase F (PNGase F) 623:656 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 7 61 theme hydrodynamic 1059:1070 arg1 mass 1072:1075 The large hydrodynamic mass 1049:1075 The large hydrodynamic mass of these oligosaccharides 1049:1101 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 62 theme oligosaccharides 1086:1101 arg1 mass 1072:1075 The large hydrodynamic mass 1049:1075 The large hydrodynamic mass of these oligosaccharides 1049:1101 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 1 63 gly glycosylation 290:302 arg2 sites 304:308 eleven consensus N-linked glycosylation sites 264:308 eleven consensus N-linked glycosylation sites 264:308 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 1 63 gly glycosylation 290:302 arg2 eleven 264:269 eleven 264:269 eleven 264:269 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 4 64 from kDa 697:699 arg1 weight 681:686 the molecular weight 667:686 the molecular weight from 110 kDa 667:699 Treatment with the endoglycosidase, peptide-N-glycosidase F (PNGase F), reduced the molecular weight from 110 kDa to 75 kDa. 8962717 1 65 theme p170 216:219 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 2 66 theme mannose 406:412 arg1 combination 386:396 a combination 384:396 a combination of high mannose and complex chains 384:431 When expressed in Chinese hamster ovary cells this was glycosylated with a combination of high mannose and complex chains. 8962717 3 67 theme spectrometric 502:514 arg1 analysis 516:523 mass spectrometric analysis 497:523 mass spectrometric analysis 497:523 The latter chains were shown by chromatographic separation and mass spectrometric analysis of tryptic digests to be clustered in the EGF-binding domain. 8962717 7 68 theme glycosylation 1202:1214 arg1 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 68 theme glycosylation 1202:1214 arg1 sites 1216:1220 consensus glycosylation sites 1192:1220 consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4 1192:1292 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 1 69 theme epidermal 221:229 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 8962717 0 70 theme domain 60:65 arg1 patterns 30:37 the glycosylation patterns 12:37 the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor 12:105 Analysis of the glycosylation patterns of the extracellular domain of the epidermal growth factor receptor expressed in Chinese hamster ovary fibroblasts. 8962717 7 71 from difference 1178:1187 arg1 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 71 from difference 1178:1187 arg1 sites 1216:1220 consensus glycosylation sites 1192:1220 consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4 1192:1292 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 72 theme proteins 1225:1232 arg1 proteins 1225:1232 proteins 1225:1232 proteins related to p170 1225:1248 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 7 72 theme proteins 1225:1232 arg1 sites 1216:1220 consensus glycosylation sites 1192:1220 consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4 1192:1292 The large hydrodynamic mass of these oligosaccharides could influence ligand binding, an effect which is likely to vary with the difference in consensus glycosylation sites of proteins related to p170 i.e. p185erbB2/neu, p180erbB3 and p180erbB4. 8962717 6 73 theme tetra-antennary 971:985 arg1 oligosaccharides 987:1002 tetra-antennary oligosaccharides 971:1002 tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc 971:1046 The data were consistent with the complex chains being trisialylated tetra-antennary oligosaccharides fucosylated on the reducing terminal GlcNAc. 8962717 1 74 theme growth 231:236 arg1 receptor 251:258 the p170 epidermal growth factor (EGF) receptor 212:258 the p170 epidermal growth factor (EGF) receptor 212:258 The extracellular domain (621 N-terminal amino acids) of the p170 epidermal growth factor (EGF) receptor has eleven consensus N-linked glycosylation sites. 15378069 1 0 theme conformational 164:177 arg1 change 179:184 conformational change 164:184 conformational change 164:184 Integrins are important adhesion receptors in all Metazoa that transmit conformational change bidirectionally across the membrane. 15378069 7 1 from legs 1036:1039 arg1 conformation 1094:1105 a previously described low-affinity bent integrin conformation 1044:1105 a previously described low-affinity bent integrin conformation 1044:1105 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 5 2 theme beta3 771:775 arg1 I 777:777 the beta3 I and hybrid domains 767:796 I 777:777 Piston-like displacement of the alpha7-helix causes a 62 degrees reorientation between the beta3 I and hybrid domains. 15378069 6 3 theme connected 832:840 arg1 domain 875:880 the rigidly connected plexin/semaphorin/integrin (PSI) domain 820:880 the rigidly connected plexin/semaphorin/integrin (PSI) domain 820:880 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 3 4 theme crystal 348:354 arg1 structures 356:365 crystal structures 348:365 crystal structures 348:365 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 3 5 theme alpha 548:552 arg1 beta3 558:562 platelet integrin alpha(IIb)beta3 530:562 platelet integrin alpha(IIb)beta3 530:562 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 4 6 theme beta3 582:586 arg1 domain 590:595 the beta3 I domain 578:595 the beta3 I domain 578:595 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 3 7 theme fibrinogen-mimetic 490:507 arg1 therapeutics 509:520 fibrinogen-mimetic therapeutics 490:520 fibrinogen-mimetic therapeutics 490:520 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 6 8 theme beta 965:968 arg1 knees 942:946 the knees 938:946 the knees of the alpha and beta legs 938:973 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 7 9 theme cell 1173:1176 arg1 surface 1178:1184 the cell surface 1169:1184 the cell surface 1169:1184 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 6 10 theme beta3 895:899 arg1 leg 901:903 the upper beta3 leg 885:903 the upper beta3 leg 885:903 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 7 11 theme bent 1080:1083 arg1 conformation 1094:1105 a previously described low-affinity bent integrin conformation 1044:1105 a previously described low-affinity bent integrin conformation 1044:1105 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 4 12 theme metal 610:614 arg1 sites 624:628 three metal binding sites 604:628 three metal binding sites 604:628 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 4 12 theme metal 610:614 arg1 loops 642:646 associated loops 631:646 associated loops 631:646 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 4 12 theme metal 610:614 arg1 alpha1- 652:658 alpha1- 652:658 alpha1- 652:658 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 3 13 theme platelet 530:537 arg1 alpha 548:552 platelet integrin alpha 530:552 platelet integrin alpha(IIb)beta3 530:562 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 3 13 theme platelet 530:537 arg1 IIb 554:556 IIb 554:556 IIb 554:556 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 3 14 theme affinity 434:441 arg1 regulation 399:408 allosteric regulation 388:408 allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain 388:479 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 4 15 from Allostery 565:573 arg1 domain 590:595 the beta3 I domain 578:595 the beta3 I domain 578:595 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 6 16 theme upper 889:893 arg1 leg 901:903 the upper beta3 leg 885:903 the upper beta3 leg 885:903 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 7 17 theme integrin 1085:1092 arg1 conformation 1094:1105 a previously described low-affinity bent integrin conformation 1044:1105 a previously described low-affinity bent integrin conformation 1044:1105 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 3 18 theme integrin 539:546 arg1 alpha 548:552 platelet integrin alpha 530:552 platelet integrin alpha(IIb)beta3 530:562 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 3 18 theme integrin 539:546 arg1 IIb 554:556 IIb 554:556 IIb 554:556 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 4 19 theme associated 631:640 arg1 sites 624:628 three metal binding sites 604:628 three metal binding sites 604:628 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 4 19 theme associated 631:640 arg1 loops 642:646 associated loops 631:646 associated loops 631:646 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 7 20 theme described 1057:1065 arg1 conformation 1094:1105 a previously described low-affinity bent integrin conformation 1044:1105 a previously described low-affinity bent integrin conformation 1044:1105 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 3 21 theme conformation 417:428 arg1 regulation 399:408 allosteric regulation 388:408 allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain 388:479 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 0 22 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for allostery in integrins 0:42 Structural basis for allostery in integrins and binding to fibrinogen-mimetic therapeutics. 15378069 7 23 theme high-affinity 1140:1152 arg1 head 1154:1157 the high-affinity head 1136:1157 the high-affinity head far above the cell surface 1136:1184 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 7 24 theme low-affinity 1067:1078 arg1 conformation 1094:1105 a previously described low-affinity bent integrin conformation 1044:1105 a previously described low-affinity bent integrin conformation 1044:1105 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 1 25 from receptors 125:133 arg1 Metazoa 142:148 all Metazoa 138:148 all Metazoa that transmit conformational change bidirectionally across the membrane 138:220 Integrins are important adhesion receptors in all Metazoa that transmit conformational change bidirectionally across the membrane. 15378069 7 26 theme leg 1112:1114 arg1 extension 1116:1124 leg extension 1112:1124 leg extension 1112:1124 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 6 27 theme alpha 955:959 arg1 knees 942:946 the knees 938:946 the knees of the alpha and beta legs 938:973 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 1 28 theme important 106:114 arg1 receptors 125:133 important adhesion receptors 106:133 important adhesion receptors 106:133 Integrins are important adhesion receptors in all Metazoa that transmit conformational change bidirectionally across the membrane. 15378069 1 28 theme important 106:114 arg1 Integrins 92:100 Integrins 92:100 Integrins 92:100 Integrins are important adhesion receptors in all Metazoa that transmit conformational change bidirectionally across the membrane. 15378069 2 29 from head 263:266 arg1 ectodomain 293:302 the ectodomain 289:302 the ectodomain 289:302 Integrin alpha and beta subunits form a head and two long legs in the ectodomain and span the membrane. 15378069 4 30 theme binding 616:622 arg1 sites 624:628 three metal binding sites 604:628 three metal binding sites 604:628 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 4 30 theme binding 616:622 arg1 loops 642:646 associated loops 631:646 associated loops 631:646 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 4 30 theme binding 616:622 arg1 alpha1- 652:658 alpha1- 652:658 alpha1- 652:658 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 3 31 theme atomic 371:376 arg1 basis 378:382 the atomic basis 367:382 the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain 367:479 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 1 32 theme adhesion 116:123 arg1 receptors 125:133 important adhesion receptors 106:133 important adhesion receptors 106:133 Integrins are important adhesion receptors in all Metazoa that transmit conformational change bidirectionally across the membrane. 15378069 1 32 theme adhesion 116:123 arg1 Integrins 92:100 Integrins 92:100 Integrins 92:100 Integrins are important adhesion receptors in all Metazoa that transmit conformational change bidirectionally across the membrane. 15378069 3 33 theme integrin 461:468 arg1 ectodomain 470:479 the integrin ectodomain 457:479 the integrin ectodomain 457:479 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 3 34 dep conformation 417:428 arg1 the 413:415 the 413:415 the 413:415 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 7 35 from Allostery 976:984 arg1 head 993:996 the head 989:996 the head 989:996 Allostery in the head thus disrupts interaction between the legs in a previously described low-affinity bent integrin conformation, and leg extension positions the high-affinity head far above the cell surface. 15378069 6 36 from Transmission 799:810 arg1 leg 901:903 the upper beta3 leg 885:903 the upper beta3 leg 885:903 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 5 37 theme alpha7-helix 712:723 arg1 displacement 692:703 Piston-like displacement 680:703 Piston-like displacement of the alpha7-helix 680:723 Piston-like displacement of the alpha7-helix causes a 62 degrees reorientation between the beta3 I and hybrid domains. 15378069 5 38 theme degrees 737:743 arg1 reorientation 745:757 a 62 degrees reorientation 732:757 a 62 degrees reorientation between the beta3 I and hybrid domains 732:796 Piston-like displacement of the alpha7-helix causes a 62 degrees reorientation between the beta3 I and hybrid domains. 15378069 3 39 theme allosteric 388:397 arg1 regulation 399:408 allosteric regulation 388:408 allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain 388:479 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 3 40 dep define 336:341 arg1 bind 522:525 bind 522:525 bind to platelet integrin alpha(IIb)beta3 522:562 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 6 41 theme PSI 870:872 arg1 domain 875:880 the rigidly connected plexin/semaphorin/integrin (PSI) domain 820:880 the rigidly connected plexin/semaphorin/integrin (PSI) domain 820:880 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 6 42 theme plexin/semaphorin/integrin 842:867 arg1 domain 875:880 the rigidly connected plexin/semaphorin/integrin (PSI) domain 820:880 the rigidly connected plexin/semaphorin/integrin (PSI) domain 820:880 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 6 43 theme A 917:917 arg1 separation 919:928 a 70 A separation 912:928 a 70 A separation between the knees of the alpha and beta legs 912:973 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 2 44 theme beta 242:245 arg1 subunits 247:254 Integrin alpha and beta subunits 223:254 Integrin alpha and beta subunits 223:254 Integrin alpha and beta subunits form a head and two long legs in the ectodomain and span the membrane. 15378069 2 45 theme alpha 232:236 arg1 subunits 247:254 Integrin alpha and beta subunits 223:254 Integrin alpha and beta subunits 223:254 Integrin alpha and beta subunits form a head and two long legs in the ectodomain and span the membrane. 15378069 2 46 theme long 276:279 arg1 legs 281:284 two long legs 272:284 two long legs 272:284 Integrin alpha and beta subunits form a head and two long legs in the ectodomain and span the membrane. 15378069 3 47 theme ectodomain 470:479 arg1 ligand 447:452 ligand 447:452 ligand of the integrin ectodomain 447:479 Here, we define with crystal structures the atomic basis for allosteric regulation of the conformation and affinity for ligand of the integrin ectodomain, and how fibrinogen-mimetic therapeutics bind to platelet integrin alpha(IIb)beta3. 15378069 2 48 from legs 281:284 arg1 ectodomain 293:302 the ectodomain 289:302 the ectodomain 289:302 Integrin alpha and beta subunits form a head and two long legs in the ectodomain and span the membrane. 15378069 5 49 theme Piston-like 680:690 arg1 displacement 692:703 Piston-like displacement 680:703 Piston-like displacement of the alpha7-helix 680:723 Piston-like displacement of the alpha7-helix causes a 62 degrees reorientation between the beta3 I and hybrid domains. 15378069 2 50 theme Integrin 223:230 arg1 alpha 232:236 Integrin alpha 223:236 Integrin alpha 223:236 Integrin alpha and beta subunits form a head and two long legs in the ectodomain and span the membrane. 15378069 4 51 theme I 588:588 arg1 domain 590:595 the beta3 I domain 578:595 the beta3 I domain 578:595 Allostery in the beta3 I domain alters three metal binding sites, associated loops and alpha1- and alpha7-helices. 15378069 0 52 theme fibrinogen-mimetic 59:76 arg1 therapeutics 78:89 fibrinogen-mimetic therapeutics 59:89 fibrinogen-mimetic therapeutics 59:89 Structural basis for allostery in integrins and binding to fibrinogen-mimetic therapeutics. 15378069 6 53 dep alpha 955:959 arg1 the 951:953 the 951:953 the 951:953 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 6 53 dep alpha 955:959 arg1 legs 970:973 legs 970:973 legs 970:973 Transmission through the rigidly connected plexin/semaphorin/integrin (PSI) domain in the upper beta3 leg causes a 70 A separation between the knees of the alpha and beta legs. 15378069 5 54 theme hybrid 783:788 arg1 domains 790:796 the beta3 I and hybrid domains 767:796 domains 790:796 Piston-like displacement of the alpha7-helix causes a 62 degrees reorientation between the beta3 I and hybrid domains. 15378069 0 55 from allostery 21:29 arg1 integrins 34:42 integrins 34:42 integrins 34:42 Structural basis for allostery in integrins and binding to fibrinogen-mimetic therapeutics. 10022822 0 0 theme cathepsin 70:78 arg1 L 80:80 cathepsin L 70:80 cathepsin L 70:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 1 1 theme molecules 306:314 arg1 processing 328:337 antigen processing 320:337 antigen processing 320:337 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 1 1 theme molecules 306:314 arg1 maturation 279:288 maturation 279:288 maturation of MHC class II molecules 279:314 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 5 2 contain has 829:831 arg2 arrangement 861:871 a predominantly beta-strand arrangement 833:871 a predominantly beta-strand arrangement 833:871 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 5 2 contain has 829:831 arg1 subdomain 819:827 the second subdomain 808:827 the second subdomain 808:827 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 4 3 theme fragment 633:640 arg1 structure 612:620 The structure 608:620 The structure of the p41 fragment 608:640 The structure of the p41 fragment demonstrates a novel fold, consisting of two subdomains, each stabilized by disulfide bridges. 10022822 6 4 theme active 950:955 arg1 cleft 962:966 the active site cleft 946:966 the active site cleft of cathepsin L 946:981 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 4 5 theme p41 629:631 arg1 fragment 633:640 the p41 fragment 625:640 the p41 fragment 625:640 The structure of the p41 fragment demonstrates a novel fold, consisting of two subdomains, each stabilized by disulfide bridges. 10022822 8 6 theme non-selective 1438:1450 arg1 cystatins 1452:1460 the rather non-selective cystatins 1427:1460 the rather non-selective cystatins 1427:1460 This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes. 10022822 6 7 theme cathepsin 971:979 arg1 L 981:981 cathepsin L 971:981 cathepsin L 971:981 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 1 8 theme antigen 320:326 arg1 processing 328:337 antigen processing 320:337 antigen processing 320:337 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 6 9 theme convergent 1076:1085 arg1 evolution 1087:1095 convergent evolution 1076:1095 convergent evolution observed in cysteine protease inhibitors 1076:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 7 10 theme different 1152:1160 arg1 fold 1162:1165 the different fold 1148:1165 the different fold of the p41 fragment 1148:1185 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 7 11 theme fragment 1178:1185 arg1 fold 1162:1165 the different fold 1148:1165 the different fold of the p41 fragment 1148:1185 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 4 12 theme novel 657:661 arg1 fold 663:666 a novel fold 655:666 a novel fold 655:666 The structure of the p41 fragment demonstrates a novel fold, consisting of two subdomains, each stabilized by disulfide bridges. 10022822 0 13 theme structural 94:103 arg1 basis 105:109 the structural basis 90:109 the structural basis for differentiation between cathepsins L and S 90:156 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 7 14 with contacts 1209:1216 arg1 top 1227:1229 the top 1223:1229 the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites 1223:1338 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 5 15 theme alpha-helix-beta-strand 763:785 arg1 arrangement 787:797 an alpha-helix-beta-strand arrangement 760:797 an alpha-helix-beta-strand arrangement 760:797 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 5 15 theme alpha-helix-beta-strand 763:785 arg1 subdomain 747:755 The first subdomain 737:755 The first subdomain 737:755 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 6 16 theme protease 1118:1125 arg1 inhibitors 1127:1136 cysteine protease inhibitors 1109:1136 cysteine protease inhibitors 1109:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 5 17 theme second 812:817 arg1 subdomain 819:827 the second subdomain 808:827 the second subdomain 808:827 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 2 18 theme invariant 360:368 arg1 chain 370:374 the invariant chain 356:374 the invariant chain 356:374 The p41 form of the invariant chain includes a fragment which specifically inhibits cathepsin L but not S. 10022822 5 19 theme beta-strand 849:859 arg1 arrangement 861:871 a predominantly beta-strand arrangement 833:871 a predominantly beta-strand arrangement 833:871 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 8 20 dep cystatins 1452:1460 arg1 contrast 1415:1422 contrast 1415:1422 contrast 1415:1422 This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes. 10022822 5 21 theme first 741:745 arg1 arrangement 787:797 an alpha-helix-beta-strand arrangement 760:797 an alpha-helix-beta-strand arrangement 760:797 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 5 21 theme first 741:745 arg1 subdomain 747:755 The first subdomain 737:755 The first subdomain 737:755 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 6 22 theme cystatins 1025:1033 arg1 edge 1017:1020 the inhibitory edge 1002:1020 the inhibitory edge of cystatins 1002:1033 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 23 theme edge 1017:1020 arg1 reminiscent 987:997 reminiscent 987:997 reminiscent 987:997 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 2 24 theme cathepsin 424:432 arg1 L 434:434 cathepsin L 424:434 cathepsin L but not S 424:444 The p41 form of the invariant chain includes a fragment which specifically inhibits cathepsin L but not S. 10022822 7 25 theme additional 1198:1207 arg1 contacts 1209:1216 additional contacts 1198:1216 additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites 1198:1338 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 3 26 theme type-1 523:528 arg1 domains 530:536 the thyroglobulin type-1 domains 505:536 the thyroglobulin type-1 domains 505:536 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 3 27 from resolution 568:577 arg1 complex 582:588 complex 582:588 complex with cathepsin L 582:605 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 1 28 theme lysosomal 163:171 arg1 proteases 182:190 The lysosomal cysteine proteases 159:190 The lysosomal cysteine proteases cathepsins S and L 159:209 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 0 29 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L 0:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 6 30 theme fragment 928:935 arg1 shape 884:888 wedge shape 878:888 wedge shape 878:888 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 30 theme fragment 928:935 arg1 arrangement 905:915 three-loop arrangement 894:915 three-loop arrangement 894:915 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 31 theme inhibitory 1006:1015 arg1 edge 1017:1020 the inhibitory edge 1002:1020 the inhibitory edge of cystatins 1002:1033 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 3 32 theme domains 530:536 arg1 homologue 492:500 a homologue 490:500 a homologue of the thyroglobulin type-1 domains 490:536 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 3 32 theme domains 530:536 arg1 fragment 480:487 the p41 fragment 472:487 the p41 fragment 472:487 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 1 33 theme cysteine 173:180 arg1 proteases 182:190 The lysosomal cysteine proteases 159:190 The lysosomal cysteine proteases cathepsins S and L 159:209 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 0 34 theme MHC 21:23 arg1 fragment 52:59 MHC class II-associated p41 Ii fragment 21:59 MHC class II-associated p41 Ii fragment bound to cathepsin L 21:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 1 35 theme invariant 256:264 arg1 chain 266:270 the invariant chain 252:270 the invariant chain 252:270 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 8 36 theme domain 1399:1404 arg1 fold 1406:1409 the thyroglobulin type-1 domain fold 1374:1409 the thyroglobulin type-1 domain fold 1374:1409 This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes. 10022822 6 37 theme evolution 1087:1095 arg1 example 1065:1071 the first example 1055:1071 the first example of convergent evolution observed in cysteine protease inhibitors 1055:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 3 38 with complex 582:588 arg1 L 605:605 cathepsin L 595:605 cathepsin L 595:605 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 7 39 theme R-domain 1238:1245 arg1 top 1227:1229 the top 1223:1229 the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites 1223:1338 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 1 40 theme chain 266:270 arg1 degradation 237:247 the degradation 233:247 the degradation of the invariant chain 233:270 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 8 41 theme type-1 1392:1397 arg1 fold 1406:1409 the thyroglobulin type-1 domain fold 1374:1409 the thyroglobulin type-1 domain fold 1374:1409 This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes. 10022822 3 42 theme thyroglobulin 509:521 arg1 domains 530:536 the thyroglobulin type-1 domains 505:536 the thyroglobulin type-1 domains 505:536 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 3 43 theme p41 476:478 arg1 homologue 492:500 a homologue 490:500 a homologue of the thyroglobulin type-1 domains 490:536 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 3 43 theme p41 476:478 arg1 fragment 480:487 the p41 fragment 472:487 the p41 fragment 472:487 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 0 44 theme II-associated 31:43 arg1 fragment 52:59 MHC class II-associated p41 Ii fragment 21:59 MHC class II-associated p41 Ii fragment bound to cathepsin L 21:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 8 45 theme target 1496:1501 arg1 enzymes 1503:1509 their target enzymes 1490:1509 their target enzymes 1490:1509 This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes. 10022822 3 46 theme cathepsin 595:603 arg1 L 605:605 cathepsin L 595:605 cathepsin L 595:605 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 7 47 theme enzymes 1254:1260 arg1 R-domain 1238:1245 the R-domain 1234:1245 the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites 1234:1338 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 7 47 theme enzymes 1254:1260 arg1 enzymes 1254:1260 the enzymes 1250:1260 the enzymes 1250:1260 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 0 48 theme class 25:29 arg1 fragment 52:59 MHC class II-associated p41 Ii fragment 21:59 MHC class II-associated p41 Ii fragment bound to cathepsin L 21:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 7 49 theme substrate-binding 1316:1332 arg1 sites 1334:1338 the specificity-determining S2 and S1' substrate-binding sites 1277:1338 the specificity-determining S2 and S1' substrate-binding sites 1277:1338 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 2 50 theme chain 370:374 arg1 form 348:351 The p41 form 340:351 The p41 form of the invariant chain 340:374 The p41 form of the invariant chain includes a fragment which specifically inhibits cathepsin L but not S. 10022822 0 51 theme Ii 49:50 arg1 fragment 52:59 MHC class II-associated p41 Ii fragment 21:59 MHC class II-associated p41 Ii fragment bound to cathepsin L 21:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 6 52 theme three-loop 894:903 arg1 arrangement 905:915 three-loop arrangement 894:915 three-loop arrangement 894:915 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 53 dep shape 884:888 arg1 The 874:876 The 874:876 The 874:876 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 54 theme p41 924:926 arg1 fragment 928:935 the p41 fragment 920:935 the p41 fragment bound to the active site cleft of cathepsin L 920:981 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 3 55 theme crystal 451:457 arg1 structure 459:467 The crystal structure 447:467 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, 447:537 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 0 56 theme p41 45:47 arg1 fragment 52:59 MHC class II-associated p41 Ii fragment 21:59 MHC class II-associated p41 Ii fragment bound to cathepsin L 21:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 1 57 theme MHC 293:295 arg1 molecules 306:314 MHC class II molecules 293:314 MHC class II molecules 293:314 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 8 58 theme thyroglobulin 1378:1390 arg1 fold 1406:1409 the thyroglobulin type-1 domain fold 1374:1409 the thyroglobulin type-1 domain fold 1374:1409 This enables inhibitors based on the thyroglobulin type-1 domain fold, in contrast to the rather non-selective cystatins, to exhibit specificity for their target enzymes. 10022822 1 59 theme class 297:301 arg1 molecules 306:314 MHC class II molecules 293:314 MHC class II molecules 293:314 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 5 60 dep arrangement 787:797 arg1 has 829:831 has 829:831 has a predominantly beta-strand arrangement 829:871 The first subdomain is an alpha-helix-beta-strand arrangement, whereas the second subdomain has a predominantly beta-strand arrangement. 10022822 4 61 theme disulfide 718:726 arg1 bridges 728:734 disulfide bridges 718:734 disulfide bridges 718:734 The structure of the p41 fragment demonstrates a novel fold, consisting of two subdomains, each stabilized by disulfide bridges. 10022822 6 62 theme first 1059:1063 arg1 example 1065:1071 the first example 1055:1071 the first example of convergent evolution observed in cysteine protease inhibitors 1055:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 63 located observed 1097:1104 arg1 inhibitors 1127:1136 cysteine protease inhibitors 1109:1136 cysteine protease inhibitors 1109:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 63 located observed 1097:1104 arg2 evolution 1087:1095 convergent evolution 1076:1095 convergent evolution observed in cysteine protease inhibitors 1076:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 6 64 theme cysteine 1109:1116 arg1 inhibitors 1127:1136 cysteine protease inhibitors 1109:1136 cysteine protease inhibitors 1109:1136 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 7 65 theme specificity-determining 1281:1303 arg1 S2 1305:1306 specificity-determining S2 1281:1306 specificity-determining S2 1281:1306 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 0 66 dep cathepsins 139:148 arg1 L 150:150 L 150:150 L 150:150 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 0 66 dep cathepsins 139:148 arg1 cathepsins 139:148 cathepsins L and S 139:156 cathepsins L and S 139:156 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 0 66 dep cathepsins 139:148 arg1 S 156:156 S 156:156 S 156:156 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 6 67 theme L 981:981 arg1 cleft 962:966 the active site cleft 946:966 the active site cleft of cathepsin L 946:981 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 1 68 dep proteases 182:190 arg1 cathepsins 192:201 cathepsins S and L 192:209 The lysosomal cysteine proteases cathepsins S and L 159:209 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 1 68 dep proteases 182:190 arg1 L 209:209 L 209:209 L 209:209 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 1 68 dep proteases 182:190 arg1 S 203:203 S 203:203 S 203:203 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 2 69 theme p41 344:346 arg1 form 348:351 The p41 form 340:351 The p41 form of the invariant chain 340:374 The p41 form of the invariant chain includes a fragment which specifically inhibits cathepsin L but not S. 10022822 7 70 theme p41 1174:1176 arg1 fragment 1178:1185 the p41 fragment 1170:1185 the p41 fragment 1170:1185 However, the different fold of the p41 fragment results in additional contacts with the top of the R-domain of the enzymes, which defines the specificity-determining S2 and S1' substrate-binding sites. 10022822 1 71 theme crucial 216:222 arg1 roles 224:228 crucial roles 216:228 crucial roles 216:228 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 0 72 theme fragment 52:59 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L 0:80 Crystal structure of MHC class II-associated p41 Ii fragment bound to cathepsin L reveals the structural basis for differentiation between cathepsins L and S. 10022822 6 73 theme wedge 878:882 arg1 shape 884:888 wedge shape 878:888 wedge shape 878:888 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 10022822 1 74 dep cathepsins 192:201 arg1 cathepsins 192:201 cathepsins S and L 192:209 The lysosomal cysteine proteases cathepsins S and L 159:209 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 1 74 dep cathepsins 192:201 arg1 L 209:209 L 209:209 L 209:209 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 1 74 dep cathepsins 192:201 arg1 S 203:203 S 203:203 S 203:203 The lysosomal cysteine proteases cathepsins S and L play crucial roles in the degradation of the invariant chain during maturation of MHC class II molecules and antigen processing. 10022822 3 75 theme fragment 480:487 arg1 structure 459:467 The crystal structure 447:467 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, 447:537 The crystal structure of the p41 fragment, a homologue of the thyroglobulin type-1 domains, has been determined at 2.0 A resolution in complex with cathepsin L. 10022822 6 76 theme site 957:960 arg1 cleft 962:966 the active site cleft 946:966 the active site cleft of cathepsin L 946:981 The wedge shape and three-loop arrangement of the p41 fragment bound to the active site cleft of cathepsin L are reminiscent of the inhibitory edge of cystatins, thus demonstrating the first example of convergent evolution observed in cysteine protease inhibitors. 17503783 0 0 theme tumor 80:84 arg1 growth 86:91 tumor growth 80:91 tumor growth 80:91 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 5 1 theme protein 1130:1136 arg1 partner 1138:1144 the putative protein partner 1117:1144 the putative protein partner 1117:1144 Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells. 17503783 5 2 from activity 1171:1178 arg1 cells 1191:1195 healthy cells 1183:1195 healthy cells 1183:1195 Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells. 17503783 5 3 theme full-length 1094:1104 arg1 enzyme 1106:1111 the full-length enzyme 1090:1111 the full-length enzyme 1090:1111 Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells. 17503783 4 4 theme catalytic 995:1003 arg1 likely 1015:1020 likely 1015:1020 likely 1015:1020 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 4 theme catalytic 995:1003 arg1 domain 1005:1010 the catalytic domain 991:1010 the catalytic domain 991:1010 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 3 5 theme regulatory 849:858 arg1 processes 860:868 regulatory processes 849:868 regulatory processes 849:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 4 6 theme alternative 954:964 arg1 splice-variants 966:980 four alternative splice-variants 949:980 four alternative splice-variants 949:980 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 3 7 theme catalytic 647:655 arg1 domain 657:662 a catalytic domain 645:662 a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase 645:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 7 theme catalytic 647:655 arg1 domains 636:642 two closely associated domains 613:642 two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes 613:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 2 8 theme hyaluronan 291:300 arg1 turnover 302:309 normal cellular hyaluronan turnover 275:309 normal cellular hyaluronan turnover 275:309 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 3 9 theme protein-protein 816:830 arg1 interactions 832:843 protein-protein interactions 816:843 protein-protein interactions 816:843 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 4 10 dep intact 939:944 arg1 intact 939:944 intact 939:944 intact 939:944 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 10 dep intact 939:944 arg1 whereas 983:989 whereas 983:989 whereas 983:989 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 10 dep intact 939:944 arg1 fold 900:903 the fold 896:903 the fold of this unique EGF-like domain 896:934 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 3 11 theme venom 732:736 arg1 hyaluronidase 738:750 bee venom hyaluronidase 728:750 bee venom hyaluronidase 728:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 2 12 theme cellular 282:289 arg1 turnover 302:309 normal cellular hyaluronan turnover 275:309 normal cellular hyaluronan turnover 275:309 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 2 13 theme splice-variants 525:539 arg1 expression 495:504 the expression 491:504 the expression of the alternative splice-variants 491:539 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 2 14 theme cancer 351:356 arg1 proliferation 358:370 cancer proliferation 351:370 cancer proliferation 351:370 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 2 15 theme normal 275:280 arg1 turnover 302:309 normal cellular hyaluronan turnover 275:309 normal cellular hyaluronan turnover 275:309 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 2 16 theme inflammatory 391:402 arg1 diseases 404:411 inflammatory diseases 391:411 inflammatory diseases 391:411 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 1 17 theme diverse 178:184 arg1 roles 200:204 diverse physiological roles 178:204 diverse physiological roles found in all tissues and body fluids 178:241 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 2 18 theme alternative 513:523 arg1 splice-variants 525:539 the alternative splice-variants 509:539 the alternative splice-variants 509:539 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 1 19 theme physiological 186:198 arg1 roles 200:204 diverse physiological roles 178:204 diverse physiological roles found in all tissues and body fluids 178:241 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 3 20 theme bee 728:730 arg1 hyaluronidase 738:750 bee venom hyaluronidase 728:750 bee venom hyaluronidase 728:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 1 21 theme roles 200:204 arg1 hyaluronan 146:155 hyaluronan 146:155 hyaluronan 146:155 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 1 21 theme roles 200:204 arg1 polysaccharide 160:173 a polysaccharide 158:173 a polysaccharide of diverse physiological roles found in all tissues and body fluids 158:241 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 5 22 theme healthy 1183:1189 arg1 cells 1191:1195 healthy cells 1183:1195 healthy cells 1183:1195 Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells. 17503783 2 23 theme human 312:316 arg1 hyaluronidase-1 318:332 human hyaluronidase-1 312:332 human hyaluronidase-1 312:332 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 0 24 theme hyaluronan 38:47 arg1 enzyme 61:66 a hyaluronan hydrolyzing enzyme 36:66 a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis 36:108 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 0 24 theme hyaluronan 38:47 arg1 Structure 0:8 Structure 0:8 Structure of human hyaluronidase-1 0:33 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 2 25 theme advanced 448:455 arg1 stages 457:462 advanced stages 448:462 advanced stages of bladder cancer 448:480 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 3 26 theme novel 759:763 arg1 domain 775:780 a novel, EGF-like domain 757:780 domain 775:780 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 26 theme novel 759:763 arg1 domains 636:642 two closely associated domains 613:642 two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes 613:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 27 dep domains 636:642 arg1 domain 657:662 a catalytic domain 645:662 a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase 645:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 27 dep domains 636:642 arg1 domain 775:780 a novel, EGF-like domain 757:780 domain 775:780 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 27 dep domains 636:642 arg1 domains 636:642 two closely associated domains 613:642 two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes 613:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 0 28 theme human 13:17 arg1 hyaluronidase-1 19:33 human hyaluronidase-1 13:33 human hyaluronidase-1 13:33 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 4 29 theme domain 929:934 arg1 intact 939:944 intact 939:944 intact 939:944 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 29 theme domain 929:934 arg1 whereas 983:989 whereas 983:989 whereas 983:989 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 29 theme domain 929:934 arg1 fold 900:903 the fold 896:903 the fold of this unique EGF-like domain 896:934 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 2 30 from function 263:270 arg1 turnover 302:309 normal cellular hyaluronan turnover 275:309 normal cellular hyaluronan turnover 275:309 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 4 31 from intact 939:944 arg1 splice-variants 966:980 four alternative splice-variants 949:980 four alternative splice-variants 949:980 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 32 from splice-variants 966:980 arg1 intact 939:944 intact 939:944 intact 939:944 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 32 from splice-variants 966:980 arg1 whereas 983:989 whereas 983:989 whereas 983:989 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 4 32 from splice-variants 966:980 arg1 fold 900:903 the fold 896:903 the fold of this unique EGF-like domain 896:934 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 1 33 located found 206:210 arg1 tissues 219:225 all tissues 215:225 all tissues 215:225 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 1 33 located found 206:210 arg1 fluids 236:241 body fluids 231:241 body fluids 231:241 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 1 33 located found 206:210 arg2 roles 200:204 diverse physiological roles 178:204 diverse physiological roles found in all tissues and body fluids 178:241 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 0 34 theme hyaluronidase-1 19:33 arg1 Structure 0:8 Structure 0:8 Structure of human hyaluronidase-1 0:33 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 0 34 theme hyaluronidase-1 19:33 arg1 enzyme 61:66 a hyaluronan hydrolyzing enzyme 36:66 a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis 36:108 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 0 35 theme hydrolyzing 49:59 arg1 enzyme 61:66 a hyaluronan hydrolyzing enzyme 36:66 a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis 36:108 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 0 35 theme hydrolyzing 49:59 arg1 Structure 0:8 Structure 0:8 Structure of human hyaluronidase-1 0:33 Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. 17503783 5 36 theme enzymatic 1161:1169 arg1 activity 1171:1178 enzymatic activity 1161:1178 enzymatic activity in healthy cells 1161:1195 Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells. 17503783 2 37 dep function 263:270 arg1 addition 247:254 addition 247:254 addition 247:254 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 4 38 theme EGF-like 920:927 arg1 domain 929:934 this unique EGF-like domain 908:934 this unique EGF-like domain 908:934 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 1 39 theme Mammalian 111:119 arg1 hyaluronidases 121:134 Mammalian hyaluronidases 111:134 Mammalian hyaluronidases 111:134 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 2 40 theme bladder 467:473 arg1 cancer 475:480 bladder cancer 467:480 bladder cancer 467:480 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 3 41 theme beta/alpha 689:698 arg1 barrel 702:707 a distorted (beta/alpha)8 barrel 676:707 a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase 676:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 42 theme EGF-like 766:773 arg1 domain 775:780 a novel, EGF-like domain 757:780 domain 775:780 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 42 theme EGF-like 766:773 arg1 domains 636:642 two closely associated domains 613:642 two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes 613:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 43 from involvement 801:811 arg1 interactions 832:843 protein-protein interactions 816:843 protein-protein interactions 816:843 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 43 from involvement 801:811 arg1 processes 860:868 regulatory processes 849:868 regulatory processes 849:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 44 theme associated 625:634 arg1 domain 657:662 a catalytic domain 645:662 a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase 645:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 44 theme associated 625:634 arg1 domain 775:780 a novel, EGF-like domain 757:780 domain 775:780 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 44 theme associated 625:634 arg1 domains 636:642 two closely associated domains 613:642 two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes 613:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 1 45 theme body 231:234 arg1 fluids 236:241 body fluids 231:241 body fluids 231:241 Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. 17503783 4 46 theme unique 913:918 arg1 domain 929:934 this unique EGF-like domain 908:934 this unique EGF-like domain 908:934 The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. 17503783 3 47 theme distorted 678:686 arg1 barrel 702:707 a distorted (beta/alpha)8 barrel 676:707 a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase 676:750 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 48 theme crystal 564:570 arg1 structure 572:580 The crystal structure 560:580 The crystal structure 560:580 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 49 theme characteristic 783:796 arg1 domain 775:780 a novel, EGF-like domain 757:780 domain 775:780 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 3 49 theme characteristic 783:796 arg1 domains 636:642 two closely associated domains 613:642 two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes 613:868 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 17503783 2 50 theme cancer 475:480 arg1 stages 457:462 advanced stages 448:462 advanced stages of bladder cancer 448:480 In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. 17503783 5 51 theme putative 1121:1128 arg1 partner 1138:1144 the putative protein partner 1117:1144 the putative protein partner 1117:1144 Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells. 17503783 3 52 theme involvement 801:811 arg1 characteristic 783:796 characteristic 783:796 characteristic 783:796 The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted (beta/alpha)8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. 22932899 6 0 theme transduction 1270:1281 arg1 mechanism 1227:1235 the mechanism 1223:1235 the mechanism of peptide processing and signal transduction 1223:1281 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 6 1 theme open 1184:1187 arg1 form 1189:1192 the open form 1180:1192 the open form 1180:1192 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 2 2 theme peptide 328:334 arg1 IV 365:366 IV 365:366 IV 365:366 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 2 2 theme peptide 328:334 arg1 hormones 336:343 processing peptide hormones 317:343 processing peptide hormones angiotensin III and IV 317:366 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 2 2 theme peptide 328:334 arg1 angiotensin 345:355 angiotensin III and IV 345:366 angiotensin 345:355 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 1 3 theme family 203:208 arg1 protein 172:178 a dimeric membrane protein 153:178 a dimeric membrane protein 153:178 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 3 theme family 203:208 arg1 member 186:191 a member 184:191 a member of the M1 family of zinc metallopeptidases 184:234 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 3 theme family 203:208 arg1 N 135:135 Human aminopeptidase N 114:135 Human aminopeptidase N (hAPN/hCD13) 114:148 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 0 4 theme novel 64:68 arg1 dimer 70:74 a novel dimer 62:74 a novel dimer 62:74 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 2 5 theme processing 317:326 arg1 IV 365:366 IV 365:366 IV 365:366 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 2 5 theme processing 317:326 arg1 hormones 336:343 processing peptide hormones 317:343 processing peptide hormones angiotensin III and IV 317:366 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 2 5 theme processing 317:326 arg1 angiotensin 345:355 angiotensin III and IV 345:366 angiotensin 345:355 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 5 6 theme complex 984:990 arg1 case 963:966 the case 959:966 the case of the bestatin complex 959:990 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 2 7 theme rennin-angiotensin 248:265 arg1 system 267:272 the rennin-angiotensin system 244:272 the rennin-angiotensin system 244:272 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 1 8 theme zinc 213:216 arg1 metallopeptidases 218:234 zinc metallopeptidases 213:234 zinc metallopeptidases 213:234 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 6 9 theme processing 1248:1257 arg1 mechanism 1227:1235 the mechanism 1223:1235 the mechanism of peptide processing and signal transduction 1223:1281 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 6 10 theme dimeric 1085:1091 arg1 member 1103:1108 a dimeric M1 family member 1083:1108 a dimeric M1 family member 1083:1108 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 3 11 theme cancer 493:498 arg1 therapy 500:506 cancer therapy 493:506 cancer therapy 493:506 In addition, hAPN is also involved in cell adhesion, endocytosis, and signal transduction and it is an important target for cancer therapy. 22932899 1 12 theme metallopeptidases 218:234 arg1 family 203:208 the M1 family 196:208 the M1 family of zinc metallopeptidases 196:234 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 6 13 theme peptide 1240:1246 arg1 processing 1248:1257 peptide processing 1240:1257 peptide processing 1240:1257 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 4 14 theme peptidomimetic 652:665 arg1 bestatin 693:700 bestatin 693:700 bestatin 693:700 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 14 theme peptidomimetic 652:665 arg1 amastatin 679:687 amastatin 679:687 amastatin 679:687 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 14 theme peptidomimetic 652:665 arg1 inhibitors 667:676 the peptidomimetic inhibitors 648:676 the peptidomimetic inhibitors 648:676 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 3 15 theme important 472:480 arg1 it 463:464 it 463:464 it 463:464 In addition, hAPN is also involved in cell adhesion, endocytosis, and signal transduction and it is an important target for cancer therapy. 22932899 3 15 theme important 472:480 arg1 target 482:487 an important target 469:487 an important target for cancer therapy 469:506 In addition, hAPN is also involved in cell adhesion, endocytosis, and signal transduction and it is an important target for cancer therapy. 22932899 5 16 theme other 781:785 arg1 enzymes 790:796 other M1 enzymes 781:796 other M1 enzymes 781:796 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 4 17 theme high 531:534 arg1 structures 561:570 the high resolution x-ray crystal structures 527:570 the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN 527:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 5 18 theme inhibitor 1016:1024 arg1 design 1026:1031 inhibitor design 1016:1031 inhibitor design 1016:1031 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 5 19 theme dimer 723:727 arg1 monomer 708:714 Each monomer 703:714 Each monomer of the dimer 703:727 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 5 19 theme dimer 723:727 arg1 dimer 723:727 the dimer 719:727 the dimer 719:727 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 2 20 dep hormones 336:343 arg1 IV 365:366 IV 365:366 IV 365:366 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 2 20 dep hormones 336:343 arg1 hormones 336:343 processing peptide hormones 317:343 processing peptide hormones angiotensin III and IV 317:366 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 2 20 dep hormones 336:343 arg1 angiotensin 345:355 angiotensin III and IV 345:366 angiotensin 345:355 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 4 21 theme hAPN 601:604 arg1 ectodomain 587:596 the dimeric ectodomain 575:596 the dimeric ectodomain of hAPN 575:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 21 theme hAPN 601:604 arg1 hAPN 601:604 hAPN 601:604 hAPN 601:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 0 22 theme X-ray 4:8 arg1 structure 18:26 The X-ray crystal structure 0:26 The X-ray crystal structure of human aminopeptidase N 0:52 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 6 23 theme signal 1263:1268 arg1 transduction 1270:1281 signal transduction 1263:1281 signal transduction 1263:1281 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 2 24 theme enzymatic 279:287 arg1 activity 289:296 its enzymatic activity 275:296 its enzymatic activity 275:296 Within the rennin-angiotensin system, its enzymatic activity is responsible for processing peptide hormones angiotensin III and IV. 22932899 6 25 theme first 1066:1070 arg1 example 1072:1078 the first example 1062:1078 the first example of a dimeric M1 family member 1062:1108 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 1 26 theme dimeric 155:161 arg1 protein 172:178 a dimeric membrane protein 153:178 a dimeric membrane protein 153:178 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 26 theme dimeric 155:161 arg1 N 135:135 Human aminopeptidase N 114:135 Human aminopeptidase N (hAPN/hCD13) 114:148 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 6 27 theme member 1103:1108 arg1 example 1072:1078 the first example 1062:1078 the first example of a dimeric M1 family member 1062:1108 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 5 28 theme catalytic 870:878 arg1 site 880:883 the catalytic site 866:883 the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering 866:947 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 1 29 theme membrane 163:170 arg1 protein 172:178 a dimeric membrane protein 153:178 a dimeric membrane protein 153:178 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 29 theme membrane 163:170 arg1 N 135:135 Human aminopeptidase N 114:135 Human aminopeptidase N (hAPN/hCD13) 114:148 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 0 30 theme crystal 10:16 arg1 structure 18:26 The X-ray crystal structure 0:26 The X-ray crystal structure of human aminopeptidase N 0:52 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 6 31 theme M1 1093:1094 arg1 member 1103:1108 a dimeric M1 family member 1083:1108 a dimeric M1 family member 1083:1108 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 0 32 theme peptide 94:100 arg1 processing 102:111 peptide processing 94:111 peptide processing 94:111 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 4 33 with complexes 614:622 arg1 bestatin 693:700 bestatin 693:700 bestatin 693:700 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 33 with complexes 614:622 arg1 amastatin 679:687 amastatin 679:687 amastatin 679:687 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 33 with complexes 614:622 arg1 inhibitors 667:676 the peptidomimetic inhibitors 648:676 the peptidomimetic inhibitors 648:676 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 33 with complexes 614:622 arg1 angiotensin 629:639 angiotensin IV 629:642 angiotensin IV 629:642 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 5 34 theme closed 766:771 arg1 form 773:776 the closed form 762:776 the closed form in other M1 enzymes and each monomer 762:813 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 0 35 theme human 31:35 arg1 N 52:52 human aminopeptidase N 31:52 human aminopeptidase N 31:52 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 6 36 from features 1138:1145 arg1 conjunction 1151:1161 conjunction 1151:1161 conjunction with a model for the open form 1151:1192 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 3 37 theme cell 407:410 arg1 adhesion 412:419 cell adhesion 407:419 cell adhesion 407:419 In addition, hAPN is also involved in cell adhesion, endocytosis, and signal transduction and it is an important target for cancer therapy. 22932899 4 38 with structures 561:570 arg1 bestatin 693:700 bestatin 693:700 bestatin 693:700 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 38 with structures 561:570 arg1 amastatin 679:687 amastatin 679:687 amastatin 679:687 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 38 with structures 561:570 arg1 inhibitors 667:676 the peptidomimetic inhibitors 648:676 the peptidomimetic inhibitors 648:676 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 38 with structures 561:570 arg1 angiotensin 629:639 angiotensin IV 629:642 angiotensin IV 629:642 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 5 39 from form 773:776 arg1 enzymes 790:796 other M1 enzymes 781:796 other M1 enzymes 781:796 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 5 39 from form 773:776 arg1 monomer 807:813 each monomer 802:813 each monomer 802:813 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 4 40 theme ectodomain 587:596 arg1 structures 561:570 the high resolution x-ray crystal structures 527:570 the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN 527:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 40 theme ectodomain 587:596 arg1 complexes 614:622 its complexes 610:622 its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin 610:700 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 0 41 theme N 52:52 arg1 structure 18:26 The X-ray crystal structure 0:26 The X-ray crystal structure of human aminopeptidase N 0:52 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 6 42 theme novel 1203:1207 arg1 insights 1209:1216 novel insights 1203:1216 novel insights into the mechanism of peptide processing and signal transduction 1203:1281 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 5 43 theme new 1003:1005 arg1 route 1007:1011 a new route 1001:1011 a new route to inhibitor design 1001:1031 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 6 44 theme structural 1127:1136 arg1 features 1138:1145 the observed structural features 1114:1145 the observed structural features 1114:1145 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 3 45 theme signal 439:444 arg1 transduction 446:457 signal transduction 439:457 signal transduction 439:457 In addition, hAPN is also involved in cell adhesion, endocytosis, and signal transduction and it is an important target for cancer therapy. 22932899 0 46 theme aminopeptidase 37:50 arg1 N 52:52 human aminopeptidase N 31:52 human aminopeptidase N 31:52 The X-ray crystal structure of human aminopeptidase N reveals a novel dimer and the basis for peptide processing. 22932899 6 47 theme observed 1118:1125 arg1 features 1138:1145 the observed structural features 1114:1145 the observed structural features 1114:1145 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 6 48 theme hAPN 1038:1041 arg1 structure 1043:1051 The hAPN structure 1034:1051 The hAPN structure 1034:1051 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 5 49 theme unique 898:903 arg1 ordering 940:947 a unique substrate/inhibitor-dependent loop ordering 896:947 the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering 866:947 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 5 50 theme internal 838:845 arg1 cavity 847:852 an internal cavity 835:852 an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering 835:947 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 4 51 theme crystal 553:559 arg1 structures 561:570 the high resolution x-ray crystal structures 527:570 the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN 527:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 5 52 theme substrate/inhibitor-dependent 905:933 arg1 ordering 940:947 a unique substrate/inhibitor-dependent loop ordering 896:947 the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering 866:947 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 1 53 theme Human 114:118 arg1 protein 172:178 a dimeric membrane protein 153:178 a dimeric membrane protein 153:178 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 53 theme Human 114:118 arg1 N 135:135 Human aminopeptidase N 114:135 Human aminopeptidase N (hAPN/hCD13) 114:148 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 53 theme Human 114:118 arg1 hAPN/hCD13 138:147 hAPN/hCD13 138:147 hAPN/hCD13 138:147 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 4 54 theme x-ray 547:551 arg1 structures 561:570 the high resolution x-ray crystal structures 527:570 the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN 527:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 5 55 theme bestatin 975:982 arg1 complex 984:990 the bestatin complex 971:990 the bestatin complex 971:990 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 5 56 theme loop 935:938 arg1 ordering 940:947 a unique substrate/inhibitor-dependent loop ordering 896:947 the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering 866:947 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 4 57 theme dimeric 579:585 arg1 ectodomain 587:596 the dimeric ectodomain 575:596 the dimeric ectodomain of hAPN 575:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 4 57 theme dimeric 579:585 arg1 hAPN 601:604 hAPN 601:604 hAPN 601:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 1 58 theme aminopeptidase 120:133 arg1 protein 172:178 a dimeric membrane protein 153:178 a dimeric membrane protein 153:178 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 58 theme aminopeptidase 120:133 arg1 N 135:135 Human aminopeptidase N 114:135 Human aminopeptidase N (hAPN/hCD13) 114:148 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 1 58 theme aminopeptidase 120:133 arg1 hAPN/hCD13 138:147 hAPN/hCD13 138:147 hAPN/hCD13 138:147 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 22932899 5 59 theme M1 787:788 arg1 enzymes 790:796 other M1 enzymes 781:796 other M1 enzymes 781:796 Each monomer of the dimer is found in what has been termed the closed form in other M1 enzymes and each monomer is characterized by an internal cavity surrounding the catalytic site as well as a unique substrate/inhibitor-dependent loop ordering, which in the case of the bestatin complex suggests a new route to inhibitor design. 22932899 4 60 theme resolution 536:545 arg1 structures 561:570 the high resolution x-ray crystal structures 527:570 the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN 527:604 Reported here are the high resolution x-ray crystal structures of the dimeric ectodomain of hAPN and its complexes with angiotensin IV and the peptidomimetic inhibitors, amastatin and bestatin. 22932899 6 61 theme family 1096:1101 arg1 member 1103:1108 a dimeric M1 family member 1083:1108 a dimeric M1 family member 1083:1108 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 6 62 with conjunction 1151:1161 arg1 model 1170:1174 a model 1168:1174 a model for the open form 1168:1192 The hAPN structure provides the first example of a dimeric M1 family member and the observed structural features, in conjunction with a model for the open form, provide novel insights into the mechanism of peptide processing and signal transduction. 22932899 1 63 theme M1 200:201 arg1 family 203:208 the M1 family 196:208 the M1 family of zinc metallopeptidases 196:234 Human aminopeptidase N (hAPN/hCD13) is a dimeric membrane protein and a member of the M1 family of zinc metallopeptidases. 12792654 8 0 theme non-catalytic 1214:1226 arg1 domain 1228:1233 this non-catalytic domain 1209:1233 this non-catalytic domain 1209:1233 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 0 1 theme Gaucher 72:78 arg1 disease 80:86 Gaucher disease 72:86 Gaucher disease 72:86 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 2 2 theme chronic 309:315 arg1 onset 317:321 early or chronic onset 300:321 early or chronic onset of severe neurological symptoms 300:353 Type 1 is characterized by hepatosplenomegaly, and types 2 and 3 by early or chronic onset of severe neurological symptoms. 12792654 5 3 theme beta/alpha 698:707 arg1 barrel 717:722 a (beta/alpha)(8) TIM barrel 695:722 a (beta/alpha)(8) TIM barrel 695:722 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 7 4 theme TIM 1020:1022 arg1 barrel 1024:1029 the TIM barrel 1016:1029 the TIM barrel 1016:1029 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 8 5 theme regulatory 1175:1184 arg1 role 1200:1203 an important regulatory or structural role 1162:1203 an important regulatory or structural role for this non-catalytic domain 1162:1233 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 4 6 from resolution 650:659 arg1 structure 618:626 the X-ray structure 608:626 the X-ray structure of GlcCerase at 2.0 A resolution 608:659 We report the X-ray structure of GlcCerase at 2.0 A resolution. 12792654 6 7 with consistent 848:857 arg1 mechanism 876:884 a catalytic mechanism 864:884 a catalytic mechanism of retention 864:897 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 5 8 theme A 779:779 arg1 clan 781:784 the glucosidase hydrolase A clan 753:784 the glucosidase hydrolase A clan 753:784 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 4 9 theme X-ray 612:616 arg1 structure 618:626 the X-ray structure 608:626 the X-ray structure of GlcCerase at 2.0 A resolution 608:659 We report the X-ray structure of GlcCerase at 2.0 A resolution. 12792654 8 10 located located 1142:1148 arg1 domain 1118:1123 a separate immunoglobulin-like domain 1087:1123 a separate immunoglobulin-like domain 1087:1123 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 8 10 located located 1142:1148 arg1 barrel 1076:1081 TIM barrel 1072:1081 TIM barrel 1072:1081 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 8 10 located located 1142:1148 arg2 L444 1134:1137 L444 1134:1137 L444 1134:1137 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 8 11 theme TIM 1072:1074 arg1 barrel 1076:1081 TIM barrel 1072:1081 TIM barrel 1072:1081 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 7 12 contain has 960:962 arg2 mutations 978:986 several other mutations 964:986 several other mutations of residues that point into the TIM barrel 964:1029 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 7 12 contain has 960:962 arg1 helix 944:948 helix 7 944:950 helix 7 944:950 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 7 12 contain has 960:962 arg1 alpha-helix 931:941 alpha-helix 931:941 alpha-helix 931:941 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 3 13 theme GlcCerase 414:422 arg1 mutations 424:432 the approximately 200 GlcCerase mutations 392:432 the approximately 200 GlcCerase mutations 392:432 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 6 14 theme catalytic 812:820 arg1 E235 831:834 E235 831:834 E235 831:834 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 6 14 theme catalytic 812:820 arg1 residues 822:829 the catalytic residues E235 and E340 808:843 the catalytic residues E235 and E340 808:843 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 6 14 theme catalytic 812:820 arg1 E340 840:843 E340 840:843 E340 840:843 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 7 15 theme residues 991:998 arg1 mutations 978:986 several other mutations 964:986 several other mutations of residues that point into the TIM barrel 964:1029 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 2 16 theme symptoms 346:353 arg1 onset 317:321 early or chronic onset 300:321 early or chronic onset of severe neurological symptoms 300:353 Type 1 is characterized by hepatosplenomegaly, and types 2 and 3 by early or chronic onset of severe neurological symptoms. 12792654 0 17 theme X-ray 0:4 arg1 enzyme 62:67 the defective enzyme 48:67 the defective enzyme in Gaucher disease 48:86 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 0 17 theme X-ray 0:4 arg1 structure 6:14 X-ray structure 0:14 X-ray structure of human acid-beta-glucosidase 0:45 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 1 18 theme Gaucher 89:95 arg1 disease 140:146 the most common lysosomal storage disease 106:146 the most common lysosomal storage disease 106:146 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 1 18 theme Gaucher 89:95 arg1 disease 97:103 Gaucher disease 89:103 Gaucher disease 89:103 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 2 19 theme neurological 333:344 arg1 symptoms 346:353 severe neurological symptoms 326:353 severe neurological symptoms 326:353 Type 1 is characterized by hepatosplenomegaly, and types 2 and 3 by early or chronic onset of severe neurological symptoms. 12792654 0 20 theme human 19:23 arg1 acid-beta-glucosidase 25:45 human acid-beta-glucosidase 19:45 human acid-beta-glucosidase 19:45 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 3 21 dep mutations 493:501 arg1 mutations 493:501 the common mutations N370S and L444P 482:517 the common mutations N370S and L444P 482:517 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 3 21 dep mutations 493:501 arg1 L444P 513:517 L444P 513:517 L444P 513:517 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 3 21 dep mutations 493:501 arg1 N370S 503:507 N370S 503:507 N370S 503:507 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 3 22 theme disease 438:444 arg1 severity 446:453 disease severity 438:453 disease severity 438:453 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 5 23 theme glucosidase 757:767 arg1 clan 781:784 the glucosidase hydrolase A clan 753:784 the glucosidase hydrolase A clan 753:784 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 9 24 theme GlcCerase 1426:1434 arg1 activity 1404:1411 the activity 1400:1411 the activity of defective GlcCerase 1400:1434 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 12792654 3 25 theme non- 538:541 arg1 neuronopathic 543:555 non- neuronopathic 538:555 non- neuronopathic 538:555 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 7 26 located located 908:914 arg2 N370 900:903 N370 900:903 N370 900:903 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 7 26 located located 908:914 arg1 helix 944:948 helix 7 944:950 helix 7 944:950 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 7 26 located located 908:914 arg1 alpha-helix 931:941 alpha-helix 931:941 alpha-helix 931:941 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 1 27 theme storage 132:138 arg1 disease 140:146 the most common lysosomal storage disease 106:146 the most common lysosomal storage disease 106:146 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 1 27 theme storage 132:138 arg1 disease 97:103 Gaucher disease 89:103 Gaucher disease 89:103 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 8 28 theme immunoglobulin-like 1098:1116 arg1 domain 1118:1123 a separate immunoglobulin-like domain 1087:1123 a separate immunoglobulin-like domain 1087:1123 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 6 29 theme retention 889:897 arg1 mechanism 876:884 a catalytic mechanism 864:884 a catalytic mechanism of retention 864:897 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 7 30 theme several 964:970 arg1 mutations 978:986 several other mutations 964:986 several other mutations of residues that point into the TIM barrel 964:1029 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 5 31 theme clan 781:784 arg1 member 743:748 a member 741:748 a member of the glucosidase hydrolase A clan 741:784 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 4 32 theme A 648:648 arg1 resolution 650:659 2.0 A resolution 644:659 2.0 A resolution 644:659 We report the X-ray structure of GlcCerase at 2.0 A resolution. 12792654 4 33 theme GlcCerase 631:639 arg1 structure 618:626 the X-ray structure 608:626 the X-ray structure of GlcCerase at 2.0 A resolution 608:659 We report the X-ray structure of GlcCerase at 2.0 A resolution. 12792654 0 34 theme acid-beta-glucosidase 25:45 arg1 enzyme 62:67 the defective enzyme 48:67 the defective enzyme in Gaucher disease 48:86 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 0 34 theme acid-beta-glucosidase 25:45 arg1 structure 6:14 X-ray structure 0:14 X-ray structure of human acid-beta-glucosidase 0:45 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 7 35 theme other 972:976 arg1 mutations 978:986 several other mutations 964:986 several other mutations of residues that point into the TIM barrel 964:1029 N370 is located on the longest alpha-helix (helix 7), which has several other mutations of residues that point into the TIM barrel. 12792654 8 36 theme important 1165:1173 arg1 role 1200:1203 an important regulatory or structural role 1162:1203 an important regulatory or structural role for this non-catalytic domain 1162:1233 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 8 37 theme structural 1189:1198 arg1 role 1200:1203 an important regulatory or structural role 1162:1203 an important regulatory or structural role for this non-catalytic domain 1162:1233 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 0 38 theme defective 52:60 arg1 enzyme 62:67 the defective enzyme 48:67 the defective enzyme in Gaucher disease 48:86 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 0 38 theme defective 52:60 arg1 structure 6:14 X-ray structure 0:14 X-ray structure of human acid-beta-glucosidase 0:45 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 8 39 theme separate 1089:1096 arg1 domain 1118:1123 a separate immunoglobulin-like domain 1087:1123 a separate immunoglobulin-like domain 1087:1123 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 0 40 from enzyme 62:67 arg1 disease 80:86 Gaucher disease 72:86 Gaucher disease 72:86 X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease. 12792654 3 41 theme neuronopathic 561:573 arg1 disease 575:581 neuronopathic disease 561:581 neuronopathic disease 561:581 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 8 42 dep barrel 1076:1081 arg1 the 1068:1070 the 1068:1070 the 1068:1070 Helix 7 is at the interface between the TIM barrel and a separate immunoglobulin-like domain on which L444 is located, suggesting an important regulatory or structural role for this non-catalytic domain. 12792654 6 43 theme catalytic 866:874 arg1 mechanism 876:884 a catalytic mechanism 864:884 a catalytic mechanism of retention 864:897 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 3 44 theme common 486:491 arg1 mutations 493:501 the common mutations N370S and L444P 482:517 the common mutations N370S and L444P 482:517 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 3 44 theme common 486:491 arg1 L444P 513:517 L444P 513:517 L444P 513:517 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 3 44 theme common 486:491 arg1 N370S 503:507 N370S 503:507 N370S 503:507 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 2 45 theme early 300:304 arg1 onset 317:321 early or chronic onset 300:321 early or chronic onset of severe neurological symptoms 300:353 Type 1 is characterized by hepatosplenomegaly, and types 2 and 3 by early or chronic onset of severe neurological symptoms. 12792654 9 46 theme engineering 1278:1288 arg1 GlcCerase 1299:1307 engineering improved GlcCerase 1278:1307 engineering improved GlcCerase 1278:1307 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 12792654 5 47 theme catalytic 666:674 arg1 domain 676:681 The catalytic domain 662:681 The catalytic domain 662:681 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 5 48 theme TIM 713:715 arg1 barrel 717:722 a (beta/alpha)(8) TIM barrel 695:722 a (beta/alpha)(8) TIM barrel 695:722 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 1 49 theme common 115:120 arg1 disease 140:146 the most common lysosomal storage disease 106:146 the most common lysosomal storage disease 106:146 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 1 49 theme common 115:120 arg1 disease 97:103 Gaucher disease 89:103 Gaucher disease 89:103 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 6 50 dep residues 822:829 arg1 E235 831:834 E235 831:834 E235 831:834 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 6 50 dep residues 822:829 arg1 residues 822:829 the catalytic residues E235 and E340 808:843 the catalytic residues E235 and E340 808:843 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 6 50 dep residues 822:829 arg1 E340 840:843 E340 840:843 E340 840:843 The distance between the catalytic residues E235 and E340 is consistent with a catalytic mechanism of retention. 12792654 1 51 from mutations 162:170 arg1 gene 179:182 the gene 175:182 the gene that encodes acid-beta-glucosidase (GlcCerase) 175:229 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 1 51 from mutations 162:170 arg1 acid-beta-glucosidase 197:217 acid-beta-glucosidase 197:217 acid-beta-glucosidase (GlcCerase) 197:229 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 1 52 theme lysosomal 122:130 arg1 disease 140:146 the most common lysosomal storage disease 106:146 the most common lysosomal storage disease 106:146 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 1 52 theme lysosomal 122:130 arg1 disease 97:103 Gaucher disease 89:103 Gaucher disease 89:103 Gaucher disease, the most common lysosomal storage disease, is caused by mutations in the gene that encodes acid-beta-glucosidase (GlcCerase). 12792654 9 53 theme GlcCerase 1299:1307 arg1 possibility 1263:1273 the possibility 1259:1273 the possibility of engineering improved GlcCerase 1259:1307 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 12792654 5 54 theme hydrolase 769:777 arg1 clan 781:784 the glucosidase hydrolase A clan 753:784 the glucosidase hydrolase A clan 753:784 The catalytic domain consists of a (beta/alpha)(8) TIM barrel, as expected for a member of the glucosidase hydrolase A clan. 12792654 2 55 theme severe 326:331 arg1 symptoms 346:353 severe neurological symptoms 326:353 severe neurological symptoms 326:353 Type 1 is characterized by hepatosplenomegaly, and types 2 and 3 by early or chronic onset of severe neurological symptoms. 12792654 9 56 theme structure-based 1359:1373 arg1 drugs 1375:1379 structure-based drugs 1359:1379 structure-based drugs aimed at restoring the activity of defective GlcCerase 1359:1434 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 12792654 3 57 theme clear 359:363 arg1 correlation 365:375 No clear correlation 356:375 No clear correlation 356:375 No clear correlation exists between the approximately 200 GlcCerase mutations and disease severity, although homozygosity for the common mutations N370S and L444P is associated with non- neuronopathic and neuronopathic disease, respectively. 12792654 9 58 theme defective 1416:1424 arg1 GlcCerase 1426:1434 defective GlcCerase 1416:1434 defective GlcCerase 1416:1434 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 12792654 9 59 theme improved 1290:1297 arg1 GlcCerase 1299:1307 engineering improved GlcCerase 1278:1307 engineering improved GlcCerase 1278:1307 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 12792654 9 60 theme enzyme-replacement 1313:1330 arg1 therapy 1332:1338 enzyme-replacement therapy 1313:1338 enzyme-replacement therapy 1313:1338 The structure provides the possibility of engineering improved GlcCerase for enzyme-replacement therapy, and for designing structure-based drugs aimed at restoring the activity of defective GlcCerase. 20877282 3 0 theme plexin 679:684 arg1 signalling 686:695 plexin signalling 679:695 plexin signalling 679:695 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 6 1 theme signalling 1369:1378 arg1 mechanism 1380:1388 a cell-cell signalling mechanism 1357:1388 a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization 1357:1440 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 1 2 theme cell 269:272 arg1 migration 274:282 cell migration 269:282 cell migration 269:282 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 7 3 theme common 1733:1738 arg1 mode 1740:1743 a common mode 1731:1743 a common mode of interaction 1731:1758 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 2 4 theme signalling 432:441 arg1 capable 414:420 capable 414:420 capable 414:420 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 1 5 theme tissues 194:200 arg1 morphogenesis 172:184 morphogenesis 172:184 morphogenesis 172:184 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 1 5 theme tissues 194:200 arg1 homeostasis 156:166 homeostasis 156:166 homeostasis 156:166 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 4 6 theme human 862:866 arg1 ecto 887:890 ecto 887:890 ecto 887:890 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 6 theme human 862:866 arg1 -SEMA4D 879:885 human PLXNB1(1-2)-SEMA4D 862:885 human PLXNB1(1-2)-SEMA4D(ecto) 862:891 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 6 theme human 862:866 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 3 7 theme structural 539:548 arg1 information 550:560 any structural information 535:560 any structural information on the plexin ectodomain 535:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 1 8 with interaction 101:111 arg1 receptors 125:133 plexin receptors 118:133 plexin receptors 118:133 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 4 9 theme murine 897:902 arg1 -Sema6A 915:921 murine PlxnA2(1-4)-Sema6A 897:921 murine PlxnA2(1-4)-Sema6A(ecto) 897:927 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 9 theme murine 897:902 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 9 theme murine 897:902 arg1 ecto 923:926 ecto 923:926 ecto 923:926 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 10 with complexes 768:776 arg1 -Sema6A 915:921 murine PlxnA2(1-4)-Sema6A 897:921 murine PlxnA2(1-4)-Sema6A(ecto) 897:927 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 10 with complexes 768:776 arg1 -SEMA4D 879:885 human PLXNB1(1-2)-SEMA4D 862:885 human PLXNB1(1-2)-SEMA4D(ecto) 862:891 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 10 with complexes 768:776 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 7 11 theme complexes 1579:1587 arg1 architecture 1559:1570 the shared generic architecture 1540:1570 the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, 1540:1715 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 0 12 theme semaphorin-plexin 20:36 arg1 signalling 38:47 semaphorin-plexin signalling 20:47 semaphorin-plexin signalling 20:47 Structural basis of semaphorin-plexin signalling. 20877282 7 13 theme semaphorin 1694:1703 arg1 plexin 1709:1714 plexin 1709:1714 plexin 1709:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 13 theme semaphorin 1694:1703 arg1 domains 1678:1684 the amino-terminal seven-bladed β-propeller (sema) domains 1627:1684 the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1627:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 13 theme semaphorin 1694:1703 arg1 semaphorin 1694:1703 semaphorin 1694:1703 semaphorin 1694:1703 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 14 theme generic 1551:1557 arg1 architecture 1559:1570 the shared generic architecture 1540:1570 the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, 1540:1715 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 6 15 theme previous 1501:1508 arg1 data 1521:1524 previous functional data 1501:1524 previous functional data 1501:1524 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 2 16 theme plexin 478:483 arg1 classes 485:491 the two largest plexin classes 462:491 the two largest plexin classes 462:491 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 3 17 with interaction 594:604 arg1 semaphorins 611:621 semaphorins 611:621 semaphorins 611:621 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 4 18 theme complexes 768:776 arg1 structures 746:755 crystal structures 738:755 crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 738:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 1 19 theme semaphorin 74:83 arg1 ligands 85:91 semaphorin ligands 74:91 semaphorin ligands 74:91 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 7 20 theme seven-bladed 1646:1657 arg1 plexin 1709:1714 plexin 1709:1714 plexin 1709:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 20 theme seven-bladed 1646:1657 arg1 domains 1678:1684 the amino-terminal seven-bladed β-propeller (sema) domains 1627:1684 the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1627:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 20 theme seven-bladed 1646:1657 arg1 semaphorin 1694:1703 semaphorin 1694:1703 semaphorin 1694:1703 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 2 21 dep classes 387:393 arg1 6 402:402 6 402:402 6 402:402 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 2 21 dep classes 387:393 arg1 4 396:396 4 396:396 4 396:396 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 7 22 theme binding 1890:1896 arg1 specificity 1898:1908 binding specificity 1890:1908 binding specificity 1890:1908 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 4 23 theme crystal 738:744 arg1 structures 746:755 crystal structures 738:755 crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 738:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 3 24 from interaction 594:604 arg1 ectodomain 576:585 the plexin ectodomain 565:585 the plexin ectodomain 565:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 0 25 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of semaphorin-plexin signalling 0:47 Structural basis of semaphorin-plexin signalling. 20877282 7 26 theme domains 1872:1878 arg1 insertions 1824:1833 distinct insertions 1815:1833 distinct insertions within or between blades of the sema domains 1815:1878 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 2 27 theme direct 425:430 arg1 signalling 432:441 direct signalling 425:441 direct signalling through members of the two largest plexin classes, B and A 425:500 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 1 28 dep homeostasis 156:166 arg1 the 152:154 the 152:154 the 152:154 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 5 29 theme resulting 1225:1233 arg1 complex 1248:1254 the resulting bivalent 2:2 complex 1221:1254 the resulting bivalent 2:2 complex 1221:1254 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 1 30 theme immune 288:293 arg1 responses 295:303 immune responses 288:303 immune responses 288:303 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 4 31 theme unliganded 936:945 arg1 structures 947:956 unliganded structures 936:956 unliganded structures of PlxnA2(1-4) and Sema6A(ecto) 936:988 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 3 32 theme extracellular 627:639 arg1 specificity 641:651 extracellular specificity 627:651 extracellular specificity 627:651 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 5 33 theme 2:2 1244:1246 arg1 complex 1248:1254 the resulting bivalent 2:2 complex 1221:1254 the resulting bivalent 2:2 complex 1221:1254 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 0 34 theme signalling 38:47 arg1 basis 11:15 Structural basis 0:15 Structural basis of semaphorin-plexin signalling 0:47 Structural basis of semaphorin-plexin signalling. 20877282 5 35 theme wild-type 1058:1066 arg1 proteins 1079:1086 wild-type and mutant proteins 1058:1086 wild-type and mutant proteins 1058:1086 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 2 36 theme vertebrate 346:355 arg1 classes 387:393 two diverse vertebrate, membrane-spanning semaphorin classes 334:393 classes 387:393 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 5 37 theme mutant 1072:1077 arg1 proteins 1079:1086 wild-type and mutant proteins 1058:1086 wild-type and mutant proteins 1058:1086 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 7 38 theme based 1791:1795 arg1 signalling 1797:1806 all semaphorin-plexin based signalling 1769:1806 all semaphorin-plexin based signalling 1769:1806 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 4 39 theme regions 804:810 arg1 complexes 768:776 cognate complexes 760:776 cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 760:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 5 40 theme plexin 1142:1147 arg1 molecules 1149:1157 two plexin molecules 1138:1157 two plexin molecules 1138:1157 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 4 41 theme plexins 815:821 arg1 regions 804:810 the semaphorin-binding regions 781:810 the semaphorin-binding regions of plexins B1 and A2 781:831 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 6 42 theme semaphorin-stabilized 1400:1420 arg1 dimerization 1429:1440 semaphorin-stabilized plexin dimerization 1400:1440 semaphorin-stabilized plexin dimerization 1400:1440 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 1 43 theme neural 240:245 arg1 connectivity 247:258 neural connectivity 240:258 neural connectivity 240:258 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 7 44 theme interaction 1748:1758 arg1 mode 1740:1743 a common mode 1731:1743 a common mode of interaction 1731:1758 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 4 45 theme Sema6A 977:982 arg1 structures 947:956 unliganded structures 936:956 unliganded structures of PlxnA2(1-4) and Sema6A(ecto) 936:988 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 2 46 theme membrane-spanning 358:374 arg1 classes 387:393 two diverse vertebrate, membrane-spanning semaphorin classes 334:393 classes 387:393 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 3 47 dep specificity 641:651 arg1 the 623:625 the 623:625 the 623:625 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 4 48 theme PLXNB1 868:873 arg1 ecto 887:890 ecto 887:890 ecto 887:890 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 48 theme PLXNB1 868:873 arg1 -SEMA4D 879:885 human PLXNB1(1-2)-SEMA4D 862:885 human PLXNB1(1-2)-SEMA4D(ecto) 862:891 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 48 theme PLXNB1 868:873 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 6 49 theme cell-cell 1359:1367 arg1 mechanism 1380:1388 a cell-cell signalling mechanism 1357:1388 a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization 1357:1440 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 1 50 theme many 189:192 arg1 tissues 194:200 many tissues 189:200 many tissues 189:200 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 4 51 theme semaphorin 838:847 arg1 -Sema6A 915:921 murine PlxnA2(1-4)-Sema6A 897:921 murine PlxnA2(1-4)-Sema6A(ecto) 897:927 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 51 theme semaphorin 838:847 arg1 -SEMA4D 879:885 human PLXNB1(1-2)-SEMA4D 862:885 human PLXNB1(1-2)-SEMA4D(ecto) 862:891 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 51 theme semaphorin 838:847 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 7 52 theme plexin 1709:1714 arg1 plexin 1709:1714 plexin 1709:1714 plexin 1709:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 52 theme plexin 1709:1714 arg1 domains 1678:1684 the amino-terminal seven-bladed β-propeller (sema) domains 1627:1684 the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1627:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 52 theme plexin 1709:1714 arg1 semaphorin 1694:1703 semaphorin 1694:1703 semaphorin 1694:1703 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 6 53 with consistent 1485:1494 arg1 data 1521:1524 previous functional data 1501:1524 previous functional data 1501:1524 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 4 54 theme PlxnA2 904:909 arg1 -Sema6A 915:921 murine PlxnA2(1-4)-Sema6A 897:921 murine PlxnA2(1-4)-Sema6A(ecto) 897:927 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 54 theme PlxnA2 904:909 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 54 theme PlxnA2 904:909 arg1 ecto 923:926 ecto 923:926 ecto 923:926 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 3 55 theme information 550:560 arg1 absence 524:530 the absence 520:530 the absence of any structural information on the plexin ectodomain 520:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 3 55 theme information 550:560 arg1 interaction 594:604 its interaction 590:604 its interaction with semaphorins 590:621 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 7 56 theme shared 1544:1549 arg1 architecture 1559:1570 the shared generic architecture 1540:1570 the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, 1540:1715 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 3 57 from absence 524:530 arg1 ectodomain 576:585 the plexin ectodomain 565:585 the plexin ectodomain 565:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 1 58 from role 232:235 arg1 cancer 261:266 cancer 261:266 cancer 261:266 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 1 58 from role 232:235 arg1 migration 274:282 cell migration 269:282 cell migration 269:282 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 1 58 from role 232:235 arg1 responses 295:303 immune responses 288:303 immune responses 288:303 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 1 58 from role 232:235 arg1 connectivity 247:258 neural connectivity 240:258 neural connectivity 240:258 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 6 59 theme functional 1510:1519 arg1 data 1521:1524 previous functional data 1501:1524 previous functional data 1501:1524 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 2 60 theme classes 485:491 arg1 A 500:500 A 500:500 A 500:500 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 2 60 theme classes 485:491 arg1 B 494:494 B 494:494 B 494:494 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 2 60 theme classes 485:491 arg1 members 451:457 members 451:457 members of the two largest plexin classes 451:491 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 5 61 theme cellular 1039:1046 arg1 assays 1048:1053 biophysical and cellular assays 1023:1053 biophysical and cellular assays of wild-type and mutant proteins 1023:1086 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 7 62 theme domains 1678:1684 arg1 contacts 1615:1622 conserved contacts 1605:1622 conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1605:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 2 63 theme largest 470:476 arg1 classes 485:491 the two largest plexin classes 462:491 the two largest plexin classes 462:491 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 7 64 theme sema 1672:1675 arg1 plexin 1709:1714 plexin 1709:1714 plexin 1709:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 64 theme sema 1672:1675 arg1 domains 1678:1684 the amino-terminal seven-bladed β-propeller (sema) domains 1627:1684 the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1627:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 64 theme sema 1672:1675 arg1 semaphorin 1694:1703 semaphorin 1694:1703 semaphorin 1694:1703 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 4 65 theme cognate 760:766 arg1 complexes 768:776 cognate complexes 760:776 cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 760:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 3 66 from ectodomain 576:585 arg1 absence 524:530 the absence 520:530 the absence of any structural information on the plexin ectodomain 520:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 3 66 from ectodomain 576:585 arg1 interaction 594:604 its interaction 590:604 its interaction with semaphorins 590:621 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 5 67 theme complex 1248:1254 arg1 avidity 1210:1216 the avidity 1206:1216 the avidity of the resulting bivalent 2:2 complex 1206:1254 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 1 68 theme ligands 85:91 arg1 signalling 60:69 Cell-cell signalling 50:69 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors 50:133 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 7 69 theme β-propeller 1659:1669 arg1 plexin 1709:1714 plexin 1709:1714 plexin 1709:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 69 theme β-propeller 1659:1669 arg1 domains 1678:1684 the amino-terminal seven-bladed β-propeller (sema) domains 1627:1684 the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1627:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 69 theme β-propeller 1659:1669 arg1 semaphorin 1694:1703 semaphorin 1694:1703 semaphorin 1694:1703 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 5 70 theme monomeric 1257:1265 arg1 semaphorin 1267:1276 monomeric semaphorin 1257:1276 monomeric semaphorin 1257:1276 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 3 71 with absence 524:530 arg1 semaphorins 611:621 semaphorins 611:621 semaphorins 611:621 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 5 72 theme biophysical 1023:1033 arg1 assays 1048:1053 biophysical and cellular assays 1023:1053 biophysical and cellular assays of wild-type and mutant proteins 1023:1086 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 7 73 theme sema 1867:1870 arg1 domains 1872:1878 the sema domains 1863:1878 the sema domains 1863:1878 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 74 theme amino-terminal 1631:1644 arg1 plexin 1709:1714 plexin 1709:1714 plexin 1709:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 74 theme amino-terminal 1631:1644 arg1 domains 1678:1684 the amino-terminal seven-bladed β-propeller (sema) domains 1627:1684 the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1627:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 7 74 theme amino-terminal 1631:1644 arg1 semaphorin 1694:1703 semaphorin 1694:1703 semaphorin 1694:1703 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 4 75 theme PlxnA2 961:966 arg1 structures 947:956 unliganded structures 936:956 unliganded structures of PlxnA2(1-4) and Sema6A(ecto) 936:988 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 3 76 from information 550:560 arg1 ectodomain 576:585 the plexin ectodomain 565:585 the plexin ectodomain 565:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 4 77 dep ectodomains 849:859 arg1 -Sema6A 915:921 murine PlxnA2(1-4)-Sema6A 897:921 murine PlxnA2(1-4)-Sema6A(ecto) 897:927 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 77 dep ectodomains 849:859 arg1 ecto 887:890 ecto 887:890 ecto 887:890 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 77 dep ectodomains 849:859 arg1 -SEMA4D 879:885 human PLXNB1(1-2)-SEMA4D 862:885 human PLXNB1(1-2)-SEMA4D(ecto) 862:891 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 77 dep ectodomains 849:859 arg1 ectodomains 849:859 semaphorin ectodomains 838:859 semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)) 838:928 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 77 dep ectodomains 849:859 arg1 ecto 923:926 ecto 923:926 ecto 923:926 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 1 78 theme plexin 118:123 arg1 receptors 125:133 plexin receptors 118:133 plexin receptors 118:133 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 5 79 dep fails 1295:1299 arg1 trigger 1304:1310 trigger 1304:1310 fails to trigger signalling 1295:1321 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 5 80 theme bivalent 1235:1242 arg1 complex 1248:1254 the resulting bivalent 2:2 complex 1221:1254 the resulting bivalent 2:2 complex 1221:1254 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 5 81 theme semaphorin 1101:1110 arg1 dimers 1112:1117 semaphorin dimers 1101:1117 semaphorin dimers 1101:1117 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 7 82 theme conserved 1605:1613 arg1 contacts 1615:1622 conserved contacts 1605:1622 conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin 1605:1714 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 4 83 dep plexins 815:821 arg1 plexins 815:821 plexins B1 and A2 815:831 plexins B1 and A2 815:831 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 83 dep plexins 815:821 arg1 A2 830:831 A2 830:831 A2 830:831 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 4 83 dep plexins 815:821 arg1 B1 823:824 B1 823:824 B1 823:824 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 2 84 theme diverse 338:344 arg1 classes 387:393 two diverse vertebrate, membrane-spanning semaphorin classes 334:393 classes 387:393 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 20877282 5 85 theme proteins 1079:1086 arg1 assays 1048:1053 biophysical and cellular assays 1023:1053 biophysical and cellular assays of wild-type and mutant proteins 1023:1086 These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). 20877282 7 86 theme distinct 1815:1822 arg1 insertions 1824:1833 distinct insertions 1815:1833 distinct insertions within or between blades of the sema domains 1815:1878 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 3 87 theme plexin 569:574 arg1 ectodomain 576:585 the plexin ectodomain 565:585 the plexin ectodomain 565:585 In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. 20877282 4 88 theme semaphorin-binding 785:802 arg1 regions 804:810 the semaphorin-binding regions 781:810 the semaphorin-binding regions of plexins B1 and A2 781:831 Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). 20877282 6 89 theme plexin 1422:1427 arg1 dimerization 1429:1440 semaphorin-stabilized plexin dimerization 1400:1440 semaphorin-stabilized plexin dimerization 1400:1440 In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. 20877282 1 90 theme Cell-cell 50:58 arg1 signalling 60:69 Cell-cell signalling 50:69 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors 50:133 Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. 20877282 7 91 theme semaphorin-plexin 1773:1789 arg1 signalling 1797:1806 all semaphorin-plexin based signalling 1769:1806 all semaphorin-plexin based signalling 1769:1806 Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed β-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity. 20877282 2 92 theme semaphorin 376:385 arg1 classes 387:393 two diverse vertebrate, membrane-spanning semaphorin classes 334:393 classes 387:393 SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. 16476442 0 0 theme enzyme 76:81 arg1 domain 27:32 the N domain 21:32 the N domain of human somatic angiotensin I-converting enzyme 21:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 0 0 theme enzyme 76:81 arg1 enzyme 76:81 human somatic angiotensin I-converting enzyme 37:81 human somatic angiotensin I-converting enzyme 37:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 6 1 theme inhibitors 1153:1162 arg1 design 1103:1108 the design 1099:1108 the design of "domain-specific" second-generation ACE inhibitors 1099:1162 The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors. 16476442 1 2 theme somatic 155:161 arg1 regulator 211:219 a key regulator 205:219 a key regulator of blood pressure 205:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 2 theme somatic 155:161 arg1 sACE 196:199 sACE 196:199 sACE 196:199 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 2 theme somatic 155:161 arg1 enzyme 188:193 Human somatic angiotensin I-converting enzyme 149:193 Human somatic angiotensin I-converting enzyme (sACE) 149:200 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 0 3 theme I-converting 63:74 arg1 enzyme 76:81 human somatic angiotensin I-converting enzyme 37:81 human somatic angiotensin I-converting enzyme 37:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 5 4 theme domain 1035:1040 arg1 cooperativity 1042:1054 the domain cooperativity 1031:1054 the domain cooperativity 1031:1054 In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. 16476442 1 5 theme angiotensin 163:173 arg1 regulator 211:219 a key regulator 205:219 a key regulator of blood pressure 205:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 5 theme angiotensin 163:173 arg1 sACE 196:199 sACE 196:199 sACE 196:199 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 5 theme angiotensin 163:173 arg1 enzyme 188:193 Human somatic angiotensin I-converting enzyme 149:193 Human somatic angiotensin I-converting enzyme (sACE) 149:200 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 6 6 theme " 1129:1129 arg1 inhibitors 1153:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors. 16476442 4 7 theme ACE 642:644 arg1 structure 622:630 the structure 618:630 the structure of testis ACE (C domain) 618:655 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 1 8 theme important 246:254 arg1 target 261:266 an important drug target 243:266 an important drug target for combating cardiovascular and renal disease 243:313 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 4 9 theme domain 705:710 arg1 structure 686:694 the crystal structure 674:694 the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function 674:887 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 1 10 theme drug 256:259 arg1 target 261:266 an important drug target 243:266 an important drug target for combating cardiovascular and renal disease 243:313 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 4 11 theme drug 778:781 arg1 lisinopril 783:792 the antihypertensive drug lisinopril 757:792 the antihypertensive drug lisinopril 757:792 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 3 12 dep peptides 475:482 arg1 peptides 475:482 the two hemoregulatory peptides 452:482 the two hemoregulatory peptides angiotensin I and bradykinin 452:511 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 3 12 dep peptides 475:482 arg1 bradykinin 502:511 bradykinin 502:511 bradykinin 502:511 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 3 12 dep peptides 475:482 arg1 I 496:496 angiotensin I 484:496 angiotensin I 484:496 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 1 13 theme I-converting 175:186 arg1 regulator 211:219 a key regulator 205:219 a key regulator of blood pressure 205:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 13 theme I-converting 175:186 arg1 sACE 196:199 sACE 196:199 sACE 196:199 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 13 theme I-converting 175:186 arg1 enzyme 188:193 Human somatic angiotensin I-converting enzyme 149:193 Human somatic angiotensin I-converting enzyme (sACE) 149:200 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 0 14 theme structural 94:103 arg1 basis 105:109 a structural basis 92:109 a structural basis for domain-specific inhibitor design 92:146 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 4 15 theme antihypertensive 761:776 arg1 lisinopril 783:792 the antihypertensive drug lisinopril 757:792 the antihypertensive drug lisinopril 757:792 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 16 theme sACE 715:718 arg1 sACE 715:718 sACE 715:718 sACE 715:718 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 16 theme sACE 715:718 arg1 domain 705:710 the N domain 699:710 the N domain of sACE 699:718 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 17 theme testis 635:640 arg1 ACE 642:644 testis ACE 635:644 testis ACE (C domain) 635:655 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 17 theme testis 635:640 arg1 domain 649:654 C domain 647:654 C domain 647:654 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 18 theme C 647:647 arg1 ACE 642:644 testis ACE 635:644 testis ACE (C domain) 635:655 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 18 theme C 647:647 arg1 domain 649:654 C domain 647:654 C domain 647:654 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 0 19 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme 0:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 4 20 theme lisinopril 783:792 arg1 absence 746:752 absence 746:752 absence 746:752 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 20 theme lisinopril 783:792 arg1 presence 733:740 presence 733:740 presence 733:740 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 0 21 theme domain-specific 115:129 arg1 design 141:146 domain-specific inhibitor design 115:146 domain-specific inhibitor design 115:146 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 6 22 theme domain-specific 1114:1128 arg1 inhibitors 1153:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors. 16476442 4 23 theme N 703:703 arg1 sACE 715:718 sACE 715:718 sACE 715:718 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 23 theme N 703:703 arg1 domain 705:710 the N domain 699:710 the N domain of sACE 699:718 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 6 24 theme ACE 1149:1151 arg1 inhibitors 1153:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors. 16476442 5 25 theme relative 973:980 arg1 positions 989:997 relative domain positions 973:997 relative domain positions for sACE that may contribute to the domain cooperativity 973:1054 In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. 16476442 2 26 theme metallopeptidase 346:361 arg1 domains 363:369 two homologous metallopeptidase domains 331:369 two homologous metallopeptidase domains 331:369 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 2 26 theme metallopeptidase 346:361 arg1 C 378:378 C 378:378 C 378:378 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 2 26 theme metallopeptidase 346:361 arg1 N 372:372 N 372:372 N 372:372 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 2 27 theme inter-domain 394:405 arg1 linker 407:412 an inter-domain linker 391:412 an inter-domain linker 391:412 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 0 28 theme domain 27:32 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme 0:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 5 29 theme domain 982:987 arg1 positions 989:997 relative domain positions 973:997 relative domain positions for sACE that may contribute to the domain cooperativity 973:1054 In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. 16476442 3 30 theme substrates 566:575 arg1 range 551:555 a range 549:555 a range of other substrates and inhibitors 549:590 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 0 31 theme N 25:25 arg1 domain 27:32 the N domain 21:32 the N domain of human somatic angiotensin I-converting enzyme 21:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 0 31 theme N 25:25 arg1 enzyme 76:81 human somatic angiotensin I-converting enzyme 37:81 human somatic angiotensin I-converting enzyme 37:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 0 32 theme inhibitor 131:139 arg1 design 141:146 domain-specific inhibitor design 115:146 domain-specific inhibitor design 115:146 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 6 33 theme second-generation 1131:1147 arg1 inhibitors 1153:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 "domain-specific" second-generation ACE inhibitors 1113:1162 The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors. 16476442 1 34 theme key 207:209 arg1 regulator 211:219 a key regulator 205:219 a key regulator of blood pressure 205:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 34 theme key 207:209 arg1 enzyme 188:193 Human somatic angiotensin I-converting enzyme 149:193 Human somatic angiotensin I-converting enzyme (sACE) 149:200 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 0 35 theme human 37:41 arg1 enzyme 76:81 human somatic angiotensin I-converting enzyme 37:81 human somatic angiotensin I-converting enzyme 37:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 1 36 theme renal 301:305 arg1 disease 307:313 renal disease 301:313 renal disease 301:313 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 5 37 theme inter-domain 932:943 arg1 linker 945:950 the inter-domain linker 928:950 the inter-domain linker 928:950 In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. 16476442 2 38 theme homologous 335:344 arg1 domains 363:369 two homologous metallopeptidase domains 331:369 two homologous metallopeptidase domains 331:369 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 2 38 theme homologous 335:344 arg1 C 378:378 C 378:378 C 378:378 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 2 38 theme homologous 335:344 arg1 N 372:372 N 372:372 N 372:372 sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. 16476442 4 39 dep presence 733:740 arg1 the 729:731 the 729:731 the 729:731 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 3 40 theme other 560:564 arg1 substrates 566:575 other substrates 560:575 other substrates 560:575 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 3 41 theme angiotensin 484:494 arg1 peptides 475:482 the two hemoregulatory peptides 452:482 the two hemoregulatory peptides angiotensin I and bradykinin 452:511 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 3 41 theme angiotensin 484:494 arg1 I 496:496 angiotensin I 484:496 angiotensin I 484:496 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 4 42 dep domain 705:710 arg1 both 721:724 both 721:724 both 721:724 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 4 43 theme crystal 678:684 arg1 structure 686:694 the crystal structure 674:694 the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function 674:887 Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. 16476442 3 44 theme hemoregulatory 460:473 arg1 peptides 475:482 the two hemoregulatory peptides 452:482 the two hemoregulatory peptides angiotensin I and bradykinin 452:511 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 3 44 theme hemoregulatory 460:473 arg1 bradykinin 502:511 bradykinin 502:511 bradykinin 502:511 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 3 44 theme hemoregulatory 460:473 arg1 I 496:496 angiotensin I 484:496 angiotensin I 484:496 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 0 45 theme angiotensin 51:61 arg1 enzyme 76:81 human somatic angiotensin I-converting enzyme 37:81 human somatic angiotensin I-converting enzyme 37:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 3 46 theme inhibitors 581:590 arg1 range 551:555 a range 549:555 a range of other substrates and inhibitors 549:590 Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. 16476442 1 47 theme blood 224:228 arg1 pressure 230:237 blood pressure 224:237 blood pressure 224:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 0 48 theme somatic 43:49 arg1 enzyme 76:81 human somatic angiotensin I-converting enzyme 37:81 human somatic angiotensin I-converting enzyme 37:81 Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. 16476442 5 49 theme most 920:923 arg1 structure 907:915 the structure 903:915 the structure of most of the inter-domain linker 903:950 In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. 16476442 1 50 theme Human 149:153 arg1 regulator 211:219 a key regulator 205:219 a key regulator of blood pressure 205:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 50 theme Human 149:153 arg1 sACE 196:199 sACE 196:199 sACE 196:199 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 50 theme Human 149:153 arg1 enzyme 188:193 Human somatic angiotensin I-converting enzyme 149:193 Human somatic angiotensin I-converting enzyme (sACE) 149:200 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 51 theme pressure 230:237 arg1 regulator 211:219 a key regulator 205:219 a key regulator of blood pressure 205:237 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 51 theme pressure 230:237 arg1 target 261:266 an important drug target 243:266 an important drug target for combating cardiovascular and renal disease 243:313 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 16476442 1 51 theme pressure 230:237 arg1 enzyme 188:193 Human somatic angiotensin I-converting enzyme 149:193 Human somatic angiotensin I-converting enzyme (sACE) 149:200 Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. 11336649 2 0 theme supershift 363:372 arg1 assays 374:379 electrophoretic mobility-shift and supershift assays 328:379 assays 374:379 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 1 theme Kell 1143:1146 arg1 transcript 1148:1157 Kell transcript 1143:1157 Kell transcript 1143:1157 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 2 2 theme non-erythroid 615:627 arg1 activity 629:636 a basal non-erythroid activity 607:636 a basal non-erythroid activity in HeLa cells 607:650 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 2 theme non-erythroid 615:627 arg1 up-regulation 658:670 (ii) up-regulation 653:670 (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context 653:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 3 theme GATA-1/Ku70/80 835:848 arg1 factors 850:856 the human GATA-1/Ku70/80 factors 825:856 the human GATA-1/Ku70/80 factors 825:856 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 5 4 theme brain 1517:1521 arg1 tissues 1523:1529 brain tissues 1517:1529 brain tissues 1517:1529 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 5 4 theme brain 1517:1521 arg1 testis 1532:1537 testis 1532:1537 testis 1532:1537 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 5 4 theme brain 1517:1521 arg1 tissues 1552:1558 lymphoid tissues 1543:1558 lymphoid tissues 1543:1558 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 2 5 theme erythroleukaemia 277:292 arg1 K562 294:297 human erythroleukaemia K562 271:297 human erythroleukaemia K562 271:297 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 6 theme amount 1133:1138 arg1 presence 1115:1122 the presence 1111:1122 the presence of a low amount of Kell transcript 1111:1157 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 3 7 theme basal 1176:1180 arg1 expression 1182:1191 basal expression 1176:1191 basal expression of the Kell protein in these cells 1176:1226 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 3 8 from expression 1182:1191 arg1 cells 1222:1226 these cells 1216:1226 these cells 1216:1226 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 7 9 theme testis 1722:1727 arg1 cells 1709:1713 the Sertoli cells 1697:1713 the Sertoli cells of the testis 1697:1727 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 2 10 theme HeLa 314:317 arg1 cells 319:323 epithelial HeLa cells 303:323 epithelial HeLa cells 303:323 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 7 11 located detected 1661:1668 arg2 protein 1644:1650 The Kell protein 1635:1650 The Kell protein 1635:1650 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 7 11 located detected 1661:1668 arg1 cells 1709:1713 the Sertoli cells 1697:1713 the Sertoli cells of the testis 1697:1727 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 1 12 theme erythroid 191:199 arg1 tissue 201:206 erythroid tissue 191:206 erythroid tissue 191:206 The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. 11336649 3 13 theme Kell 1200:1203 arg1 protein 1205:1211 the Kell protein 1196:1211 the Kell protein 1196:1211 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 2 14 theme K562 577:580 arg1 cells 582:586 K562 cells 577:586 K562 cells 577:586 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 1 15 theme Kell 113:116 arg1 protein 170:176 a protein 168:176 a protein expressed in erythroid tissue only 168:211 The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. 11336649 1 15 theme Kell 113:116 arg1 antigen 130:136 The Kell blood-group antigen 109:136 The Kell blood-group antigen 109:136 The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. 11336649 6 16 theme Kell 1617:1620 arg1 transcripts 1622:1632 Kell transcripts 1617:1632 Kell transcripts 1617:1632 Moreover, most tissues analysed exhibited low levels of Kell transcripts. 11336649 2 17 theme promoter 689:696 arg1 activity 698:705 the 5' distal promoter activity 675:705 the 5' distal promoter activity occurs only in the erythroid context 675:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 18 theme promoter 250:257 arg1 activity 259:266 the KEL promoter activity 242:266 the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells 242:323 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 19 from K562 294:297 arg1 analysis 230:237 Transcriptional analysis 214:237 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis 214:476 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 20 theme HeLa 988:991 arg1 cells 993:997 HeLa cells 988:997 HeLa cells 988:997 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 21 theme erythroid 726:734 arg1 context 736:742 the erythroid context 722:742 the erythroid context 722:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 0 22 theme protein 52:58 arg1 expression 60:69 Kell protein expression 47:69 Kell protein expression 47:69 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 2 23 theme KEL 518:520 arg1 promoter 522:529 the KEL promoter 514:529 the KEL promoter 514:529 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 4 24 theme erythroid 1372:1380 arg1 tissue 1382:1387 erythroid tissue 1372:1387 erythroid tissue 1372:1387 DNA sequencing of the transcript revealed a sequence identical to that obtained from erythroid tissue. 11336649 6 25 theme low 1603:1605 arg1 levels 1607:1612 low levels 1603:1612 low levels of Kell transcripts 1603:1632 Moreover, most tissues analysed exhibited low levels of Kell transcripts. 11336649 2 26 from cells 319:323 arg1 analysis 230:237 Transcriptional analysis 214:237 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis 214:476 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 27 theme KEL 1015:1017 arg1 expression 1019:1028 KEL expression 1015:1028 KEL expression in HeLa cells 1015:1042 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 7 28 theme Kell 1639:1642 arg1 protein 1644:1650 The Kell protein 1635:1650 The Kell protein 1635:1650 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 2 29 theme strong 542:547 arg1 activity 565:572 a strong transcriptional activity 540:572 a strong transcriptional activity in K562 cells 540:586 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 5 30 theme KEL 1408:1410 arg1 expression 1412:1421 KEL expression 1408:1421 KEL expression 1408:1421 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 2 31 theme co-transfection 424:438 arg1 studies 440:446 co-transfection studies 424:446 co-transfection studies 424:446 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 6 32 theme most 1571:1574 arg1 tissues 1576:1582 most tissues 1571:1582 most tissues analysed 1571:1591 Moreover, most tissues analysed exhibited low levels of Kell transcripts. 11336649 8 33 theme erythroid 1910:1918 arg1 tissue 1920:1925 erythroid tissue 1910:1925 erythroid tissue 1910:1925 Altogether, the results indicated that KEL expression is not restricted to erythroid tissue. 11336649 0 34 theme Transcriptional 0:14 arg1 regulation 16:25 Transcriptional regulation 0:25 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 0:107 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 2 35 dep activity 698:705 arg1 occurs 707:712 occurs 707:712 occurs only in the erythroid context 707:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 5 36 theme Kell 1489:1492 arg1 mRNAs 1494:1498 Kell mRNAs 1489:1498 Kell mRNAs 1489:1498 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 2 37 located localized 766:774 arg2 motifs 759:764 a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs 607:764 a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity 607:916 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 37 located localized 766:774 arg1 region 790:795 the exon 1 region 779:795 the exon 1 region 779:795 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 38 from activity 259:266 arg1 K562 294:297 human erythroleukaemia K562 271:297 human erythroleukaemia K562 271:297 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 38 from activity 259:266 arg1 cells 319:323 epithelial HeLa cells 303:323 epithelial HeLa cells 303:323 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 39 from activity 629:636 arg1 cells 646:650 HeLa cells 641:650 HeLa cells 641:650 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 0 40 theme KEL 34:36 arg1 gene 38:41 the KEL gene 30:41 the KEL gene 30:41 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 2 41 theme promoter 900:907 arg1 activity 909:916 the promoter activity 896:916 the promoter activity 896:916 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 42 theme electrophoretic 328:342 arg1 mobility-shift 344:357 electrophoretic mobility-shift and supershift assays 328:379 mobility-shift 344:357 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 7 43 theme follicular 1807:1816 arg1 cells 1828:1832 the follicular dendritic cells 1803:1832 the follicular dendritic cells 1803:1832 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 2 44 theme chloramphenicol 382:396 arg1 assays 416:421 chloramphenicol acetyltransferase assays 382:421 chloramphenicol acetyltransferase assays 382:421 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 45 theme low 1129:1131 arg1 amount 1133:1138 a low amount 1127:1138 a low amount of Kell transcript 1127:1157 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 3 45 theme low 1129:1131 arg1 transcript 1148:1157 Kell transcript 1143:1157 Kell transcript 1143:1157 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 5 46 theme lymphoid 1543:1550 arg1 tissues 1523:1529 brain tissues 1517:1529 brain tissues 1517:1529 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 5 46 theme lymphoid 1543:1550 arg1 tissues 1552:1558 lymphoid tissues 1543:1558 lymphoid tissues 1543:1558 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 4 47 theme DNA 1287:1289 arg1 sequencing 1291:1300 DNA sequencing 1287:1300 DNA sequencing of the transcript 1287:1318 DNA sequencing of the transcript revealed a sequence identical to that obtained from erythroid tissue. 11336649 2 48 theme activity 629:636 arg1 motifs 759:764 a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs 607:764 a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity 607:916 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 7 49 theme lymphoid 1736:1743 arg1 tissues 1745:1751 lymphoid tissues 1736:1751 lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells 1736:1832 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 5 50 theme human 1393:1397 arg1 tissues 1399:1405 human tissues 1393:1405 human tissues 1393:1405 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 3 51 theme transcript 1148:1157 arg1 amount 1133:1138 a low amount 1127:1138 a low amount of Kell transcript 1127:1157 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 3 51 theme transcript 1148:1157 arg1 transcript 1148:1157 Kell transcript 1143:1157 Kell transcript 1143:1157 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 2 52 theme basal 609:613 arg1 activity 629:636 a basal non-erythroid activity 607:636 a basal non-erythroid activity in HeLa cells 607:650 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 52 theme basal 609:613 arg1 up-regulation 658:670 (ii) up-regulation 653:670 (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context 653:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 53 theme human 829:833 arg1 factors 850:856 the human GATA-1/Ku70/80 factors 825:856 the human GATA-1/Ku70/80 factors 825:856 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 54 theme human 271:275 arg1 K562 294:297 human erythroleukaemia K562 271:297 human erythroleukaemia K562 271:297 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 55 theme HeLa 641:644 arg1 cells 646:650 HeLa cells 641:650 HeLa cells 641:650 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 4 56 theme identical 1340:1348 arg1 sequence 1331:1338 a sequence 1329:1338 a sequence identical to that obtained from erythroid tissue 1329:1387 DNA sequencing of the transcript revealed a sequence identical to that obtained from erythroid tissue. 11336649 2 57 theme epithelial 303:312 arg1 cells 319:323 epithelial HeLa cells 303:323 epithelial HeLa cells 303:323 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 58 theme two 755:757 arg1 motifs 759:764 a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs 607:764 a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity 607:916 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 8 59 theme KEL 1874:1876 arg1 expression 1878:1887 KEL expression 1874:1887 KEL expression 1874:1887 Altogether, the results indicated that KEL expression is not restricted to erythroid tissue. 11336649 3 60 theme protein 1205:1211 arg1 expression 1182:1191 basal expression 1176:1191 basal expression of the Kell protein in these cells 1176:1226 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 2 61 theme repressing 966:975 arg1 factors 977:983 the repressing factors 962:983 the repressing factors 962:983 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 4 62 theme transcript 1309:1318 arg1 sequencing 1291:1300 DNA sequencing 1287:1300 DNA sequencing of the transcript 1287:1318 DNA sequencing of the transcript revealed a sequence identical to that obtained from erythroid tissue. 11336649 2 63 theme Transcriptional 214:228 arg1 analysis 230:237 Transcriptional analysis 214:237 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis 214:476 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 64 from expression 1019:1028 arg1 cells 1038:1042 HeLa cells 1033:1042 HeLa cells 1033:1042 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 6 65 theme transcripts 1622:1632 arg1 levels 1607:1612 low levels 1603:1612 low levels of Kell transcripts 1603:1632 Moreover, most tissues analysed exhibited low levels of Kell transcripts. 11336649 2 66 theme activity 698:705 arg1 up-regulation 658:670 (ii) up-regulation 653:670 (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context 653:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 66 theme activity 698:705 arg1 activity 629:636 a basal non-erythroid activity 607:636 a basal non-erythroid activity in HeLa cells 607:650 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 67 theme activity 259:266 arg1 analysis 230:237 Transcriptional analysis 214:237 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis 214:476 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 1 68 theme blood-group 118:128 arg1 protein 170:176 a protein 168:176 a protein expressed in erythroid tissue only 168:211 The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. 11336649 1 68 theme blood-group 118:128 arg1 antigen 130:136 The Kell blood-group antigen 109:136 The Kell blood-group antigen 109:136 The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. 11336649 7 69 theme Sertoli 1701:1707 arg1 cells 1709:1713 the Sertoli cells 1697:1713 the Sertoli cells of the testis 1697:1727 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 2 70 from activity 565:572 arg1 cells 582:586 K562 cells 577:586 K562 cells 577:586 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 71 theme Western-blot 1264:1275 arg1 analysis 1277:1284 Western-blot analysis 1264:1284 Western-blot analysis 1264:1284 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 0 72 theme expression 60:69 arg1 regulation 16:25 Transcriptional regulation 0:25 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 0:107 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 2 73 theme KEL 246:248 arg1 activity 259:266 the KEL promoter activity 242:266 the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells 242:323 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 7 74 located localized 1790:1798 arg1 cells 1828:1832 the follicular dendritic cells 1803:1832 the follicular dendritic cells 1803:1832 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 7 74 located localized 1790:1798 arg2 spleen 1758:1763 spleen 1758:1763 spleen 1758:1763 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 0 75 theme erythroid 74:82 arg1 cells 102:106 erythroid and non-erythroid cells 74:106 erythroid and non-erythroid cells 74:106 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 0 76 from regulation 16:25 arg1 cells 102:106 erythroid and non-erythroid cells 74:106 erythroid and non-erythroid cells 74:106 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 5 77 theme dot-blot 1443:1450 arg1 analysis 1452:1459 dot-blot analysis 1443:1459 dot-blot analysis 1443:1459 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 0 78 theme non-erythroid 88:100 arg1 cells 102:106 erythroid and non-erythroid cells 74:106 erythroid and non-erythroid cells 74:106 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 2 79 dep up-regulation 658:670 arg1 ii 654:655 ii 654:655 ii 654:655 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 80 theme transcriptional 549:563 arg1 activity 565:572 a strong transcriptional activity 540:572 a strong transcriptional activity in K562 cells 540:586 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 81 theme promoter 941:948 arg1 activity 950:957 the promoter activity 937:957 the promoter activity by the repressing factors in HeLa cells 937:997 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 81 theme promoter 941:948 arg1 incomplete 1003:1012 incomplete 1003:1012 incomplete 1003:1012 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 82 theme distal 682:687 arg1 activity 698:705 the 5' distal promoter activity 675:705 the 5' distal promoter activity occurs only in the erythroid context 675:742 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 83 theme HeLa 1033:1036 arg1 cells 1038:1042 HeLa cells 1033:1042 HeLa cells 1033:1042 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 2 84 from analysis 230:237 arg1 K562 294:297 human erythroleukaemia K562 271:297 human erythroleukaemia K562 271:297 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 84 from analysis 230:237 arg1 cells 319:323 epithelial HeLa cells 303:323 epithelial HeLa cells 303:323 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 85 theme following 491:499 arg1 results 501:507 the following results 487:507 the following results 487:507 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 86 theme acetyltransferase 398:414 arg1 assays 416:421 chloramphenicol acetyltransferase assays 382:421 chloramphenicol acetyltransferase assays 382:421 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 3 87 theme primer-extension 1069:1084 arg1 analysis 1086:1093 primer-extension analysis 1069:1093 primer-extension analysis 1069:1093 KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. 11336649 2 88 from inhibition 923:932 arg1 cells 582:586 K562 cells 577:586 K562 cells 577:586 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 5 89 theme mRNAs 1494:1498 arg1 levels 1479:1484 high levels 1474:1484 high levels of Kell mRNAs 1474:1498 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 2 90 theme site-directed 452:464 arg1 mutagenesis 466:476 site-directed mutagenesis 452:476 site-directed mutagenesis 452:476 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 0 91 theme gene 38:41 arg1 regulation 16:25 Transcriptional regulation 0:25 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 0:107 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 2 92 theme activity 909:916 arg1 down-regulation 877:891 down-regulation 877:891 down-regulation of the promoter activity 877:916 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 0 93 theme Kell 47:50 arg1 expression 60:69 Kell protein expression 47:69 Kell protein expression 47:69 Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells. 11336649 7 94 theme dendritic 1818:1826 arg1 cells 1828:1832 the follicular dendritic cells 1803:1832 the follicular dendritic cells 1803:1832 The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. 11336649 2 95 from activity 950:957 arg1 cells 993:997 HeLa cells 988:997 HeLa cells 988:997 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 5 96 theme high 1474:1477 arg1 levels 1479:1484 high levels 1474:1484 high levels of Kell mRNAs 1474:1498 In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. 11336649 2 97 dep exhibits 531:538 arg1 i 511:511 i 511:511 i 511:511 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 98 theme exon 783:786 arg1 region 790:795 the exon 1 region 779:795 the exon 1 region 779:795 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 11336649 2 99 from motifs 759:764 arg1 cells 582:586 K562 cells 577:586 K562 cells 577:586 Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. 20008117 6 0 theme low-temperature 864:878 arg1 incubation 880:889 low-temperature incubation 864:889 low-temperature incubation 864:889 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 6 1 theme common 773:778 arg1 CFTR 806:809 DeltaF508 CFTR 796:809 DeltaF508 CFTR 796:809 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 6 1 theme common 773:778 arg1 mutation 780:787 The most common mutation 764:787 The most common mutation in CF 764:793 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 2 2 theme mutant 322:327 arg1 CFTR 329:332 wild-type and mutant CFTR 308:332 wild-type and mutant CFTR in its native environment 308:358 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 2 3 from trafficking 293:303 arg1 environment 348:358 its native environment 337:358 its native environment 337:358 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 7 4 theme apical 1096:1101 arg1 membrane 1103:1110 the apical membrane 1092:1110 the apical membrane 1092:1110 Incubation with small-molecule correctors resulted in DeltaF508 CFTR at the apical membrane but did not restore apical stability. 20008117 10 5 theme native 1762:1767 arg1 environment 1769:1779 its native environment 1758:1779 its native environment 1758:1779 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 6 6 theme endoplasmic 829:839 arg1 reticulum 841:849 endoplasmic reticulum 829:849 endoplasmic reticulum retention 829:859 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 1 7 theme epithelial 164:173 arg1 cells 175:179 epithelial cells 164:179 epithelial cells 164:179 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 8 8 theme mutant 1167:1172 arg1 protein 1174:1180 the mutant protein 1163:1180 the mutant protein 1163:1180 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 7 9 theme DeltaF508 1074:1082 arg1 CFTR 1084:1087 DeltaF508 CFTR 1074:1087 DeltaF508 CFTR 1074:1087 Incubation with small-molecule correctors resulted in DeltaF508 CFTR at the apical membrane but did not restore apical stability. 20008117 9 10 theme CFTR 1480:1483 arg1 properties 1466:1475 intrinsic properties 1456:1475 intrinsic properties of CFTR that may affect its apical stability 1456:1520 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 9 11 theme surface 1711:1717 arg1 CFTR 1719:1722 surface CFTR 1711:1722 surface CFTR 1711:1722 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 8 12 theme inhibitor 1232:1240 arg1 Dynasore 1242:1249 the dynamin inhibitor Dynasore 1220:1249 the dynamin inhibitor Dynasore 1220:1249 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 3 13 theme intracellular 515:527 arg1 compartments 529:540 intracellular compartments 515:540 intracellular compartments 515:540 We labeled the apical pool of CFTR and subsequently visualized the protein in intracellular compartments. 20008117 8 14 theme apical 1189:1194 arg1 membrane 1196:1203 the apical membrane 1185:1203 the apical membrane 1185:1203 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 6 15 theme apical 914:919 arg1 membrane 921:928 the apical membrane 910:928 the apical membrane 910:928 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 6 16 from mutation 780:787 arg1 CF 792:793 CF 792:793 CF 792:793 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 7 17 theme small-molecule 1036:1049 arg1 correctors 1051:1060 small-molecule correctors 1036:1060 small-molecule correctors 1036:1060 Incubation with small-molecule correctors resulted in DeltaF508 CFTR at the apical membrane but did not restore apical stability. 20008117 10 18 theme protein 1814:1820 arg1 properties 1796:1805 properties 1796:1805 properties of the protein and modulation of endocytic trafficking 1796:1860 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 1 19 theme cystic 200:205 arg1 CF 217:218 CF 217:218 CF 217:218 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 1 19 theme cystic 200:205 arg1 fibrosis 207:214 cystic fibrosis 200:214 cystic fibrosis (CF) 200:219 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 9 20 dep necessary 1572:1580 arg1 influenced 1684:1693 influenced 1684:1693 influenced the turnover of surface CFTR 1684:1722 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 5 21 theme CFTR 642:645 arg1 endocytosis 647:657 CFTR endocytosis 642:657 CFTR endocytosis 642:657 CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. 20008117 0 22 from cultures 100:107 arg1 Modulation 0:9 Modulation 0:9 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 0:108 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 10 23 theme endocytic 1840:1848 arg1 trafficking 1850:1860 endocytic trafficking 1840:1860 endocytic trafficking 1840:1860 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 0 24 theme CFTR 60:63 arg1 trafficking 24:34 endocytic trafficking 14:34 endocytic trafficking 14:34 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 0 24 theme CFTR 60:63 arg1 stability 47:55 apical stability 40:55 apical stability 40:55 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 10 25 theme modulation 1826:1835 arg1 properties 1796:1805 properties 1796:1805 properties of the protein and modulation of endocytic trafficking 1796:1860 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 1 26 theme apical 137:142 arg1 CFTR 110:113 CFTR 110:113 CFTR 110:113 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 1 26 theme apical 137:142 arg1 channel 153:159 a highly regulated apical chloride channel 118:159 a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF) 118:219 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 2 27 from stability 265:273 arg1 environment 348:358 its native environment 337:358 its native environment 337:358 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 8 28 theme cholesterol-extracting 1259:1280 arg1 cyclodextrin 1288:1299 the cholesterol-extracting agent cyclodextrin 1255:1299 the cholesterol-extracting agent cyclodextrin 1255:1299 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 0 29 theme primary 68:74 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 0 30 from Modulation 0:9 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 10 31 theme apical 1730:1735 arg1 stability 1737:1745 apical stability 1730:1745 apical stability of CFTR in its native environment 1730:1779 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 3 32 from protein 504:510 arg1 compartments 529:540 intracellular compartments 515:540 intracellular compartments 515:540 We labeled the apical pool of CFTR and subsequently visualized the protein in intracellular compartments. 20008117 0 33 theme airway 82:87 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 2 34 dep cultures 427:434 arg1 i.e. 361:364 i.e. 361:364 i.e. 361:364 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 0 35 from stability 47:55 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 5 36 theme HAE 714:716 arg1 cells 718:722 nonpolarized HAE cells 701:722 nonpolarized HAE cells 701:722 CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. 20008117 0 37 theme apical 40:45 arg1 stability 47:55 apical stability 40:55 apical stability 40:55 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 8 38 theme inhibitor 1376:1384 arg1 MG-132 1386:1391 the proteasomal inhibitor MG-132 1360:1391 the proteasomal inhibitor MG-132 1360:1391 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 7 39 with Incubation 1020:1029 arg1 correctors 1051:1060 small-molecule correctors 1036:1060 small-molecule correctors 1036:1060 Incubation with small-molecule correctors resulted in DeltaF508 CFTR at the apical membrane but did not restore apical stability. 20008117 2 40 theme airway 403:408 arg1 cultures 427:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 2 41 theme primary 389:395 arg1 cultures 427:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 0 42 theme endocytic 14:22 arg1 trafficking 24:34 endocytic trafficking 14:34 endocytic trafficking 14:34 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 10 43 theme CFTR 1750:1753 arg1 stability 1737:1745 apical stability 1730:1745 apical stability of CFTR in its native environment 1730:1779 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 9 44 theme apical 1603:1608 arg1 membrane 1610:1617 the apical membrane 1599:1617 the apical membrane 1599:1617 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 7 45 theme apical 1132:1137 arg1 stability 1139:1147 apical stability 1132:1147 apical stability 1132:1147 Incubation with small-molecule correctors resulted in DeltaF508 CFTR at the apical membrane but did not restore apical stability. 20008117 5 46 theme epithelial 742:751 arg1 line 758:761 a polarized epithelial cell line 730:761 a polarized epithelial cell line 730:761 CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. 20008117 2 47 from environment 348:358 arg1 stability 265:273 apical stability 258:273 apical stability 258:273 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 2 47 from environment 348:358 arg1 trafficking 293:303 intracellular trafficking 279:303 intracellular trafficking 279:303 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 6 48 theme wild-type 1004:1012 arg1 CFTR 1014:1017 wild-type CFTR 1004:1017 wild-type CFTR 1004:1017 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 8 49 theme DeltaF508 1427:1435 arg1 endocytosis 1412:1422 endocytosis 1412:1422 endocytosis of DeltaF508 1412:1435 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 0 50 theme stability 47:55 arg1 Modulation 0:9 Modulation 0:9 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 0:108 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 10 51 from stability 1737:1745 arg1 environment 1769:1779 its native environment 1758:1779 its native environment 1758:1779 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 9 52 theme efficient 1641:1649 arg1 recycling 1658:1666 efficient apical recycling 1641:1666 efficient apical recycling 1641:1666 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 4 53 theme apical 563:568 arg1 surface 570:576 the apical surface 559:576 the apical surface 559:576 CFTR moved from the apical surface to endosomes and then efficiently recycled back to the surface. 20008117 2 54 dep stability 265:273 arg1 the 254:256 the 254:256 the 254:256 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 3 55 theme CFTR 467:470 arg1 pool 459:462 the apical pool 448:462 the apical pool of CFTR 448:470 We labeled the apical pool of CFTR and subsequently visualized the protein in intracellular compartments. 20008117 9 56 theme properties 1466:1475 arg1 examination 1441:1451 examination 1441:1451 examination of intrinsic properties of CFTR that may affect its apical stability 1441:1520 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 2 57 theme CFTR 329:332 arg1 stability 265:273 apical stability 258:273 apical stability 258:273 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 2 57 theme CFTR 329:332 arg1 trafficking 293:303 intracellular trafficking 279:303 intracellular trafficking 279:303 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 6 58 theme reticulum 841:849 arg1 retention 851:859 endoplasmic reticulum retention 829:859 endoplasmic reticulum retention 829:859 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 9 59 theme N-linked 1537:1544 arg1 oligosaccharides 1546:1561 N-linked oligosaccharides 1537:1561 N-linked oligosaccharides 1537:1561 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 9 60 theme intrinsic 1456:1464 arg1 properties 1466:1475 intrinsic properties 1456:1475 intrinsic properties of CFTR that may affect its apical stability 1456:1520 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 2 61 from CFTR 329:332 arg1 environment 348:358 its native environment 337:358 its native environment 337:358 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 2 62 theme intracellular 279:291 arg1 trafficking 293:303 intracellular trafficking 279:303 intracellular trafficking 279:303 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 1 63 theme cells 175:179 arg1 CFTR 110:113 CFTR 110:113 CFTR 110:113 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 1 63 theme cells 175:179 arg1 channel 153:159 a highly regulated apical chloride channel 118:159 a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF) 118:219 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 6 64 theme endocytic 933:941 arg1 compartments 943:954 endocytic compartments 933:954 endocytic compartments 933:954 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 9 65 theme CFTR 1719:1722 arg1 turnover 1699:1706 the turnover 1695:1706 the turnover of surface CFTR 1695:1722 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 8 66 theme dynamin 1224:1230 arg1 Dynasore 1242:1249 the dynamin inhibitor Dynasore 1220:1249 the dynamin inhibitor Dynasore 1220:1249 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 2 67 theme wild-type 308:316 arg1 CFTR 329:332 wild-type and mutant CFTR 308:332 wild-type and mutant CFTR in its native environment 308:358 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 10 68 dep protein 1814:1820 arg1 the 1810:1812 the 1810:1812 the 1810:1812 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 9 69 theme apical 1505:1510 arg1 stability 1512:1520 its apical stability 1501:1520 its apical stability 1501:1520 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 10 70 theme trafficking 1850:1860 arg1 protein 1814:1820 protein 1814:1820 protein 1814:1820 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 10 70 theme trafficking 1850:1860 arg1 modulation 1826:1835 modulation 1826:1835 modulation of endocytic trafficking 1826:1860 Thus apical stability of CFTR in its native environment is affected by properties of the protein and modulation of endocytic trafficking. 20008117 8 71 theme agent 1282:1286 arg1 cyclodextrin 1288:1299 the cholesterol-extracting agent cyclodextrin 1255:1299 the cholesterol-extracting agent cyclodextrin 1255:1299 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 1 72 theme regulated 127:135 arg1 CFTR 110:113 CFTR 110:113 CFTR 110:113 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 1 72 theme regulated 127:135 arg1 channel 153:159 a highly regulated apical chloride channel 118:159 a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF) 118:219 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 0 73 from trafficking 24:34 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 0 74 theme human 76:80 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 1 75 theme chloride 144:151 arg1 CFTR 110:113 CFTR 110:113 CFTR 110:113 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 1 75 theme chloride 144:151 arg1 channel 153:159 a highly regulated apical chloride channel 118:159 a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF) 118:219 CFTR is a highly regulated apical chloride channel of epithelial cells that is mutated in cystic fibrosis (CF). 20008117 0 76 theme epithelial 89:98 arg1 cultures 100:107 primary human airway epithelial cultures 68:107 primary human airway epithelial cultures 68:107 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 2 77 theme apical 258:263 arg1 stability 265:273 apical stability 258:273 apical stability 258:273 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 8 78 theme DeltaF508 1341:1349 arg1 internalization 1322:1336 internalization 1322:1336 internalization of DeltaF508 1322:1349 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 5 79 theme nonpolarized 701:712 arg1 cells 718:722 nonpolarized HAE cells 701:722 nonpolarized HAE cells 701:722 CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. 20008117 2 80 theme epithelial 410:419 arg1 cultures 427:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 8 81 theme proteasomal 1364:1374 arg1 MG-132 1386:1391 the proteasomal inhibitor MG-132 1360:1391 the proteasomal inhibitor MG-132 1360:1391 To stabilize the mutant protein at the apical membrane, we found that the dynamin inhibitor Dynasore and the cholesterol-extracting agent cyclodextrin dramatically reduced internalization of DeltaF508, whereas the proteasomal inhibitor MG-132 completely blocked endocytosis of DeltaF508. 20008117 2 82 theme native 341:346 arg1 environment 348:358 its native environment 337:358 its native environment 337:358 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 0 83 theme trafficking 24:34 arg1 Modulation 0:9 Modulation 0:9 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 0:108 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures. 20008117 9 84 link N-linked 1537:1544 arg1 oligosaccharides 1546:1561 N-linked oligosaccharides 1537:1561 N-linked oligosaccharides 1537:1561 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 20008117 2 85 theme human 397:401 arg1 cultures 427:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 2 86 theme differentiated 374:387 arg1 cultures 427:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 5 87 theme polarized 732:740 arg1 line 758:761 a polarized epithelial cell line 730:761 a polarized epithelial cell line 730:761 CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. 20008117 5 88 theme cell 753:756 arg1 line 758:761 a polarized epithelial cell line 730:761 a polarized epithelial cell line 730:761 CFTR endocytosis occurred more slowly in polarized than in nonpolarized HAE cells or in a polarized epithelial cell line. 20008117 6 89 theme DeltaF508 796:804 arg1 CFTR 806:809 DeltaF508 CFTR 796:809 DeltaF508 CFTR 796:809 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 6 89 theme DeltaF508 796:804 arg1 mutation 780:787 The most common mutation 764:787 The most common mutation in CF 764:793 The most common mutation in CF, DeltaF508 CFTR, was rescued from endoplasmic reticulum retention by low-temperature incubation but transited from the apical membrane to endocytic compartments more rapidly and recycled less efficiently than wild-type CFTR. 20008117 2 90 theme HAE 422:424 arg1 cultures 427:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 highly differentiated primary human airway epithelial (HAE) cultures 367:434 In this study, we characterized the apical stability and intracellular trafficking of wild-type and mutant CFTR in its native environment, i.e., highly differentiated primary human airway epithelial (HAE) cultures. 20008117 3 91 theme apical 452:457 arg1 pool 459:462 the apical pool 448:462 the apical pool of CFTR 448:470 We labeled the apical pool of CFTR and subsequently visualized the protein in intracellular compartments. 20008117 9 92 theme apical 1651:1656 arg1 recycling 1658:1666 efficient apical recycling 1641:1666 efficient apical recycling 1641:1666 On examination of intrinsic properties of CFTR that may affect its apical stability, we found that N-linked oligosaccharides were not necessary for transport to the apical membrane but were required for efficient apical recycling and, therefore, influenced the turnover of surface CFTR. 3571235 4 0 theme intramembranous 481:495 arg1 domain 497:502 the intramembranous domain 477:502 the intramembranous domain (residues 36-71) of glycophorin B 477:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 0 theme intramembranous 481:495 arg1 B 536:536 glycophorin B 524:536 glycophorin B 524:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 0 theme intramembranous 481:495 arg1 residues 505:512 residues 36-71 505:518 residues 36-71 505:518 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 1 gly sialoglycoprotein 624:640 arg1 sialoglycoprotein 624:640 the major sialoglycoprotein 614:640 the major sialoglycoprotein (glycophorin A) 614:656 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 1 gly sialoglycoprotein 624:640 arg1 A 655:655 glycophorin A 643:655 glycophorin A 643:655 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 2 2 gly sialoglycoproteins 157:174 arg1 glycophorins 177:188 glycophorins B and C 177:196 glycophorins B and C 177:196 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 2 gly sialoglycoproteins 157:174 arg1 sialoglycoproteins 157:174 two sialoglycoproteins 153:174 two sialoglycoproteins (glycophorins B and C) 153:197 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 5 3 theme hydrophobic 690:700 arg1 domain 702:707 the hydrophobic domain 686:707 the hydrophobic domain (residues 49-88) of glycophorin C 686:741 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 5 3 theme hydrophobic 690:700 arg1 C 741:741 glycophorin C 729:741 glycophorin C 729:741 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 5 3 theme hydrophobic 690:700 arg1 residues 710:717 residues 49-88 710:723 residues 49-88 710:723 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 0 4 from membranes 44:52 arg1 Glycophorins 0:11 Glycophorins B and C 0:19 Glycophorins B and C from human erythrocyte membranes 0:52 Glycophorins B and C from human erythrocyte membranes. 3571235 0 4 from membranes 44:52 arg1 C 19:19 C 19:19 C 19:19 Glycophorins B and C from human erythrocyte membranes. 3571235 0 4 from membranes 44:52 arg1 B 13:13 B 13:13 B 13:13 Glycophorins B and C from human erythrocyte membranes. 3571235 2 5 dep glycophorins 177:188 arg1 glycophorins 177:188 glycophorins B and C 177:196 glycophorins B and C 177:196 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 5 dep glycophorins 177:188 arg1 C 196:196 C 196:196 C 196:196 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 5 dep glycophorins 177:188 arg1 B 190:190 B 190:190 B 190:190 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 5 6 theme domain 702:707 arg1 sequence 674:681 The amino acid sequence 659:681 The amino acid sequence 659:681 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 4 7 theme acid 460:463 arg1 similar 569:575 similar 569:575 similar 569:575 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 7 theme acid 460:463 arg1 sequence 465:472 The amino acid sequence 450:472 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B 450:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 2 8 theme permeation 273:282 arg1 chromatography 284:297 high-performance ion exchange and gel permeation chromatography 235:297 chromatography 284:297 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 4 9 theme sialoglycoprotein 624:640 arg1 region 604:609 the hydrophobic region 588:609 the hydrophobic region of the major sialoglycoprotein (glycophorin A) 588:656 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 5 10 theme glycophorin 729:739 arg1 C 741:741 glycophorin C 729:741 glycophorin C 729:741 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 1 11 theme sequence 72:79 arg1 analysis 81:88 sequence analysis 72:88 sequence analysis 72:88 Purification and sequence analysis. 3571235 4 12 theme major 618:622 arg1 sialoglycoprotein 624:640 the major sialoglycoprotein 614:640 the major sialoglycoprotein (glycophorin A) 614:656 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 12 theme major 618:622 arg1 A 655:655 glycophorin A 643:655 glycophorin A 643:655 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 2 13 theme preparative 125:135 arg1 purification 137:148 the preparative purification 121:148 the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes 121:230 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 14 theme Triton 318:323 arg1 X-100 325:329 Triton X-100 318:329 Triton X-100 318:329 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 15 from membranes 222:230 arg1 purification 137:148 the preparative purification 121:148 the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes 121:230 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 4 16 theme glycophorin 643:653 arg1 sialoglycoprotein 624:640 the major sialoglycoprotein 614:640 the major sialoglycoprotein (glycophorin A) 614:656 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 16 theme glycophorin 643:653 arg1 A 655:655 glycophorin A 643:655 glycophorin A 643:655 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 3 17 theme detectable 371:380 arg1 contaminants 382:393 any detectable contaminants 367:393 any detectable contaminants 367:393 Glycophorin B was obtained without any detectable contaminants, and glycophorin C exhibited a purity of about 90-95%. 3571235 4 18 theme B 536:536 arg1 domain 497:502 the intramembranous domain 477:502 the intramembranous domain (residues 36-71) of glycophorin B 477:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 18 theme B 536:536 arg1 B 536:536 glycophorin B 524:536 glycophorin B 524:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 18 theme B 536:536 arg1 residues 505:512 residues 36-71 505:518 residues 36-71 505:518 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 5 19 theme amino 663:667 arg1 sequence 674:681 The amino acid sequence 659:681 The amino acid sequence 659:681 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 5 20 theme acid 669:672 arg1 sequence 674:681 The amino acid sequence 659:681 The amino acid sequence 659:681 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 4 21 theme hydrophobic 592:602 arg1 region 604:609 the hydrophobic region 588:609 the hydrophobic region of the major sialoglycoprotein (glycophorin A) 588:656 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 0 22 theme human 26:30 arg1 membranes 44:52 human erythrocyte membranes 26:52 human erythrocyte membranes 26:52 Glycophorins B and C from human erythrocyte membranes. 3571235 5 23 dep residues 710:717 arg1 49-88 719:723 49-88 719:723 49-88 719:723 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 2 24 from exchange 256:263 arg1 presence 306:313 the presence 302:313 the presence of Triton X-100 302:329 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 3 25 theme % 447:447 arg1 purity 426:431 a purity 424:431 a purity of about 90-95% 424:447 Glycophorin B was obtained without any detectable contaminants, and glycophorin C exhibited a purity of about 90-95%. 3571235 5 26 theme C 741:741 arg1 domain 702:707 the hydrophobic domain 686:707 the hydrophobic domain (residues 49-88) of glycophorin C 686:741 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 5 26 theme C 741:741 arg1 C 741:741 glycophorin C 729:741 glycophorin C 729:741 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 5 26 theme C 741:741 arg1 residues 710:717 residues 49-88 710:723 residues 49-88 710:723 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 4 27 dep residues 505:512 arg1 36-71 514:518 36-71 514:518 36-71 514:518 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 28 theme amino 454:458 arg1 similar 569:575 similar 569:575 similar 569:575 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 28 theme amino 454:458 arg1 sequence 465:472 The amino acid sequence 450:472 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B 450:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 3 29 theme glycophorin 400:410 arg1 C 412:412 glycophorin C 400:412 glycophorin C 400:412 Glycophorin B was obtained without any detectable contaminants, and glycophorin C exhibited a purity of about 90-95%. 3571235 2 30 theme sialoglycoproteins 157:174 arg1 purification 137:148 the preparative purification 121:148 the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes 121:230 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 31 theme erythrocyte 210:220 arg1 membranes 222:230 human erythrocyte membranes 204:230 human erythrocyte membranes 204:230 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 0 32 theme erythrocyte 32:42 arg1 membranes 44:52 human erythrocyte membranes 26:52 human erythrocyte membranes 26:52 Glycophorins B and C from human erythrocyte membranes. 3571235 4 33 theme glycophorin 524:534 arg1 B 536:536 glycophorin B 524:536 glycophorin B 524:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 2 34 theme human 204:208 arg1 membranes 222:230 human erythrocyte membranes 204:230 human erythrocyte membranes 204:230 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 35 theme high-performance 235:250 arg1 exchange 256:263 high-performance ion exchange and gel permeation chromatography 235:297 exchange 256:263 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 36 theme gel 269:271 arg1 chromatography 284:297 high-performance ion exchange and gel permeation chromatography 235:297 chromatography 284:297 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 2 37 theme X-100 325:329 arg1 presence 306:313 the presence 302:313 the presence of Triton X-100 302:329 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 3 38 theme Glycophorin 332:342 arg1 B 344:344 Glycophorin B 332:344 Glycophorin B 332:344 Glycophorin B was obtained without any detectable contaminants, and glycophorin C exhibited a purity of about 90-95%. 3571235 2 39 theme ion 252:254 arg1 exchange 256:263 high-performance ion exchange and gel permeation chromatography 235:297 exchange 256:263 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 5 40 theme cDNA 829:832 arg1 sequencing 834:843 cDNA sequencing 829:843 cDNA sequencing 829:843 The amino acid sequence of the hydrophobic domain (residues 49-88) of glycophorin C, that was also determined, agreed completely with the structure recently deduced from cDNA sequencing. 3571235 2 41 from chromatography 284:297 arg1 presence 306:313 the presence 302:313 the presence of Triton X-100 302:329 We have developed methods for the preparative purification of two sialoglycoproteins (glycophorins B and C) from human erythrocyte membranes by high-performance ion exchange and gel permeation chromatography in the presence of Triton X-100. 3571235 4 42 theme domain 497:502 arg1 similar 569:575 similar 569:575 similar 569:575 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 4 42 theme domain 497:502 arg1 sequence 465:472 The amino acid sequence 450:472 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B 450:536 The amino acid sequence of the intramembranous domain (residues 36-71) of glycophorin B was determined and found to be similar to that of the hydrophobic region of the major sialoglycoprotein (glycophorin A). 3571235 0 43 dep Glycophorins 0:11 arg1 Glycophorins 0:11 Glycophorins B and C 0:19 Glycophorins B and C from human erythrocyte membranes 0:52 Glycophorins B and C from human erythrocyte membranes. 3571235 0 43 dep Glycophorins 0:11 arg1 C 19:19 C 19:19 C 19:19 Glycophorins B and C from human erythrocyte membranes. 3571235 0 43 dep Glycophorins 0:11 arg1 B 13:13 B 13:13 B 13:13 Glycophorins B and C from human erythrocyte membranes. 4742735 0 0 theme human 56:60 arg1 immunoglobulin 66:79 a human IgM immunoglobulin 54:79 a human IgM immunoglobulin 54:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 2 1 theme heavy 426:430 arg1 chain 432:436 each heavy chain 421:436 each heavy chain 421:436 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 1 2 theme bridges 197:203 arg1 location 171:178 the location 167:178 the location of all disulfide bridges and oligosaccharides 167:224 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 1 3 theme chain 120:124 arg1 sequence 97:104 The amino acid sequence 82:104 The amino acid sequence 82:104 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 0 4 theme immunoglobulin 66:79 arg1 chain 45:49 the Mu heavy chain 32:49 the Mu heavy chain of a human IgM immunoglobulin 32:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 1 5 theme oligosaccharides 209:224 arg1 location 171:178 the location 167:178 the location of all disulfide bridges and oligosaccharides 167:224 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 0 6 theme IgM 62:64 arg1 immunoglobulin 66:79 a human IgM immunoglobulin 54:79 a human IgM immunoglobulin 54:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 2 7 theme constant 268:275 arg1 regions 277:283 the constant regions 264:283 the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains 264:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 2 8 theme immunoglobulin 288:301 arg1 regions 277:283 the constant regions 264:283 the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains 264:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 2 9 theme epsilon 328:334 arg1 chains 342:347 epsilon heavy chains 328:347 epsilon heavy chains 328:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 0 10 theme amino 9:13 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin 0:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 1 11 theme human 131:135 arg1 immunoglobulin 141:154 a human IgM immunoglobulin 129:154 a human IgM immunoglobulin 129:154 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 0 12 theme Complete 0:7 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin 0:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 1 13 theme IgM 137:139 arg1 immunoglobulin 141:154 a human IgM immunoglobulin 129:154 a human IgM immunoglobulin 129:154 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 1 14 theme immunoglobulin 141:154 arg1 chain 120:124 the micro, chain 109:124 chain 120:124 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 0 15 theme acid 15:18 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin 0:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 2 16 theme alpha 317:321 arg1 regions 277:283 the constant regions 264:283 the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains 264:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 2 17 theme chains 342:347 arg1 regions 277:283 the constant regions 264:283 the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains 264:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 2 18 theme heavy 336:340 arg1 chains 342:347 epsilon heavy chains 328:347 epsilon heavy chains 328:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 0 19 theme heavy 39:43 arg1 chain 45:49 the Mu heavy chain 32:49 the Mu heavy chain of a human IgM immunoglobulin 32:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 1 20 theme amino 86:90 arg1 sequence 97:104 The amino acid sequence 82:104 The amino acid sequence 82:104 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 0 21 theme Mu 36:37 arg1 chain 45:49 the Mu heavy chain 32:49 the Mu heavy chain of a human IgM immunoglobulin 32:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 1 22 theme acid 92:95 arg1 sequence 97:104 The amino acid sequence 82:104 The amino acid sequence 82:104 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 2 23 theme regions 277:283 arg1 homology 252:259 The homology 248:259 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains 248:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 2 24 theme gamma 310:314 arg1 regions 277:283 the constant regions 264:283 the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains 264:347 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 2 25 theme evolutionary 357:368 arg1 relationships 370:382 evolutionary relationships 357:382 evolutionary relationships 357:382 The homology of the constant regions of immunoglobulin micro, gamma, alpha, and epsilon heavy chains reveals evolutionary relationships and suggests that two genes code for each heavy chain. 4742735 0 26 theme chain 45:49 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin 0:79 Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. 4742735 1 27 theme micro 113:117 arg1 chain 120:124 the micro, chain 109:124 chain 120:124 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 4742735 1 28 theme disulfide 187:195 arg1 bridges 197:203 all disulfide bridges 183:203 all disulfide bridges 183:203 The amino acid sequence of the micro, chain of a human IgM immunoglobulin, including the location of all disulfide bridges and oligosaccharides, has been determined. 22660477 8 0 theme B 1110:1110 arg1 receptor 1113:1120 GABA(B) receptor 1105:1120 GABA(B) receptor that is unique to the GABAergic system 1105:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 8 1 theme GABA 1105:1108 arg1 receptor 1113:1120 GABA(B) receptor 1105:1120 GABA(B) receptor that is unique to the GABAergic system 1105:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 5 2 theme GBR1 595:598 arg1 ectodomain 600:609 the GBR1 ectodomain 591:609 the GBR1 ectodomain 591:609 Here we demonstrate that the GBR2 ectodomain directly interacts with the GBR1 ectodomain to increase agonist affinity by selectively stabilizing the agonist-bound conformation of GBR1. 22660477 8 3 theme active 1089:1094 arg1 state 1096:1100 the active state 1085:1100 the active state of GABA(B) receptor that is unique to the GABAergic system 1085:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 0 4 theme GABA 74:77 arg1 GBR2 91:94 human GABA(B) receptor GBR2 68:94 human GABA(B) receptor GBR2 68:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 6 5 theme heterodimeric 786:798 arg1 interface 800:808 a polar heterodimeric interface 778:808 a polar heterodimeric interface 778:808 We present the crystal structure of the GBR2 ectodomain, which reveals a polar heterodimeric interface. 22660477 8 6 theme structural 940:949 arg1 data 966:969 our structural and functional data 936:969 our structural and functional data 936:969 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 7 7 theme heterodimer 837:847 arg1 contacts 849:856 specific heterodimer contacts 828:856 specific heterodimer contacts from both subunits 828:875 We also identify specific heterodimer contacts from both subunits, and GBR1 residues involved in ligand recognition. 22660477 4 8 theme GBR1 405:408 arg1 subunits 419:426 GBR1 and GBR2 subunits 405:426 GBR1 and GBR2 subunits 405:426 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 3 9 theme several 315:321 arg1 disorders 336:344 several neurological disorders 315:344 several neurological disorders 315:344 Malfunction of GABA(B) receptor has been implicated in several neurological disorders. 22660477 7 10 theme ligand 908:913 arg1 recognition 915:925 ligand recognition 908:925 ligand recognition 908:925 We also identify specific heterodimer contacts from both subunits, and GBR1 residues involved in ligand recognition. 22660477 4 11 theme B 352:352 arg1 assembly 393:400 a heterodimeric assembly 377:400 a heterodimeric assembly 377:400 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 4 11 theme B 352:352 arg1 receptor 355:362 GABA(B) receptor 347:362 GABA(B) receptor 347:362 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 3 12 theme neurological 323:334 arg1 disorders 336:344 several neurological disorders 315:344 several neurological disorders 315:344 Malfunction of GABA(B) receptor has been implicated in several neurological disorders. 22660477 5 13 theme GBR2 551:554 arg1 ectodomain 556:565 the GBR2 ectodomain 547:565 the GBR2 ectodomain 547:565 Here we demonstrate that the GBR2 ectodomain directly interacts with the GBR1 ectodomain to increase agonist affinity by selectively stabilizing the agonist-bound conformation of GBR1. 22660477 0 14 theme GBR2 91:94 arg1 GBR2 91:94 human GABA(B) receptor GBR2 68:94 human GABA(B) receptor GBR2 68:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 0 14 theme GBR2 91:94 arg1 domain 58:63 the extracellular domain 40:63 the extracellular domain of human GABA(B) receptor GBR2 40:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 4 15 theme subunits 419:426 arg1 assembly 393:400 a heterodimeric assembly 377:400 a heterodimeric assembly 377:400 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 4 15 theme subunits 419:426 arg1 receptor 355:362 GABA(B) receptor 347:362 GABA(B) receptor 347:362 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 8 16 theme open 1029:1032 arg1 conformation 1034:1045 a constitutively open conformation 1012:1045 a constitutively open conformation 1012:1045 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 7 17 theme specific 828:835 arg1 contacts 849:856 specific heterodimer contacts 828:856 specific heterodimer contacts from both subunits 828:875 We also identify specific heterodimer contacts from both subunits, and GBR1 residues involved in ligand recognition. 22660477 0 18 theme receptor 82:89 arg1 GBR2 91:94 human GABA(B) receptor GBR2 68:94 human GABA(B) receptor GBR2 68:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 6 19 theme ectodomain 752:761 arg1 structure 730:738 the crystal structure 718:738 the crystal structure 718:738 We present the crystal structure of the GBR2 ectodomain, which reveals a polar heterodimeric interface. 22660477 2 20 theme brain 244:248 arg1 function 250:257 mammalian brain function 234:257 mammalian brain function 234:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 3 21 theme receptor 283:290 arg1 Malfunction 260:270 Malfunction 260:270 Malfunction of GABA(B) receptor 260:290 Malfunction of GABA(B) receptor has been implicated in several neurological disorders. 22660477 6 22 theme GBR2 747:750 arg1 ectodomain 752:761 the GBR2 ectodomain 743:761 the GBR2 ectodomain 743:761 We present the crystal structure of the GBR2 ectodomain, which reveals a polar heterodimeric interface. 22660477 7 23 from subunits 868:875 arg1 residues 887:894 GBR1 residues 882:894 GBR1 residues involved in ligand recognition 882:925 We also identify specific heterodimer contacts from both subunits, and GBR1 residues involved in ligand recognition. 22660477 7 23 from subunits 868:875 arg1 contacts 849:856 specific heterodimer contacts 828:856 specific heterodimer contacts from both subunits 828:875 We also identify specific heterodimer contacts from both subunits, and GBR1 residues involved in ligand recognition. 22660477 4 24 theme G 502:502 arg1 coupling 512:519 G protein coupling 502:519 G protein coupling 502:519 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 6 25 theme polar 780:784 arg1 interface 800:808 a polar heterodimeric interface 778:808 a polar heterodimeric interface 778:808 We present the crystal structure of the GBR2 ectodomain, which reveals a polar heterodimeric interface. 22660477 8 26 theme GBR2 989:992 arg1 ectodomain 994:1003 the GBR2 ectodomain 985:1003 the GBR2 ectodomain 985:1003 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 8 27 theme GABAergic 1144:1152 arg1 system 1154:1159 the GABAergic system 1140:1159 the GABAergic system 1140:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 4 28 theme GABA 347:350 arg1 assembly 393:400 a heterodimeric assembly 377:400 a heterodimeric assembly 377:400 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 4 28 theme GABA 347:350 arg1 receptor 355:362 GABA(B) receptor 347:362 GABA(B) receptor 347:362 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 0 29 theme functional 14:23 arg1 interaction 25:35 functional interaction 14:35 functional interaction 14:35 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 2 30 theme GABA 174:177 arg1 receptor 214:221 a G protein-coupled receptor 194:221 a G protein-coupled receptor central to mammalian brain function 194:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 2 30 theme GABA 174:177 arg1 receptor 182:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 4 31 theme protein 504:510 arg1 coupling 512:519 G protein coupling 502:519 G protein coupling 502:519 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 5 32 theme agonist-bound 671:683 arg1 conformation 685:696 the agonist-bound conformation 667:696 the agonist-bound conformation of GBR1 667:704 Here we demonstrate that the GBR2 ectodomain directly interacts with the GBR1 ectodomain to increase agonist affinity by selectively stabilizing the agonist-bound conformation of GBR1. 22660477 2 33 theme metabotropic 161:172 arg1 receptor 214:221 a G protein-coupled receptor 194:221 a G protein-coupled receptor central to mammalian brain function 194:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 2 33 theme metabotropic 161:172 arg1 receptor 182:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 2 34 theme protein-coupled 198:212 arg1 receptor 214:221 a G protein-coupled receptor 194:221 a G protein-coupled receptor central to mammalian brain function 194:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 2 34 theme protein-coupled 198:212 arg1 receptor 182:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 8 35 theme functional 955:964 arg1 data 966:969 our structural and functional data 936:969 our structural and functional data 936:969 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 8 36 theme structural 1061:1070 arg1 asymmetry 1072:1080 a structural asymmetry 1059:1080 a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system 1059:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 4 37 theme GBR2 414:417 arg1 subunits 419:426 GBR1 and GBR2 subunits 405:426 GBR1 and GBR2 subunits 405:426 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 0 38 theme domain 58:63 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 0 38 theme domain 58:63 arg1 interaction 25:35 functional interaction 14:35 functional interaction 14:35 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 8 39 theme receptor 1113:1120 arg1 state 1096:1100 the active state 1085:1100 the active state of GABA(B) receptor that is unique to the GABAergic system 1085:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 5 40 theme agonist 623:629 arg1 affinity 631:638 agonist affinity 623:638 agonist affinity 623:638 Here we demonstrate that the GBR2 ectodomain directly interacts with the GBR1 ectodomain to increase agonist affinity by selectively stabilizing the agonist-bound conformation of GBR1. 22660477 6 41 theme crystal 722:728 arg1 structure 730:738 the crystal structure 718:738 the crystal structure 718:738 We present the crystal structure of the GBR2 ectodomain, which reveals a polar heterodimeric interface. 22660477 0 42 theme extracellular 44:56 arg1 GBR2 91:94 human GABA(B) receptor GBR2 68:94 human GABA(B) receptor GBR2 68:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 0 42 theme extracellular 44:56 arg1 domain 58:63 the extracellular domain 40:63 the extracellular domain of human GABA(B) receptor GBR2 40:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 3 43 theme B 280:280 arg1 receptor 283:290 GABA(B) receptor 275:290 GABA(B) receptor 275:290 Malfunction of GABA(B) receptor has been implicated in several neurological disorders. 22660477 1 44 theme Inhibitory 97:106 arg1 neurotransmission 108:124 Inhibitory neurotransmission 97:124 Inhibitory neurotransmission 97:124 Inhibitory neurotransmission is mediated primarily by GABA. 22660477 2 45 theme mammalian 234:242 arg1 function 250:257 mammalian brain function 234:257 mammalian brain function 234:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 7 46 theme GBR1 882:885 arg1 residues 887:894 GBR1 residues 882:894 GBR1 residues involved in ligand recognition 882:925 We also identify specific heterodimer contacts from both subunits, and GBR1 residues involved in ligand recognition. 22660477 8 47 from asymmetry 1072:1080 arg1 state 1096:1100 the active state 1085:1100 the active state of GABA(B) receptor that is unique to the GABAergic system 1085:1159 Lastly, our structural and functional data indicate that the GBR2 ectodomain adopts a constitutively open conformation, suggesting a structural asymmetry in the active state of GABA(B) receptor that is unique to the GABAergic system. 22660477 3 48 theme GABA 275:278 arg1 receptor 283:290 GABA(B) receptor 275:290 GABA(B) receptor 275:290 Malfunction of GABA(B) receptor has been implicated in several neurological disorders. 22660477 2 49 theme B 179:179 arg1 receptor 214:221 a G protein-coupled receptor 194:221 a G protein-coupled receptor central to mammalian brain function 194:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 2 49 theme B 179:179 arg1 receptor 182:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 5 50 theme GBR1 701:704 arg1 conformation 685:696 the agonist-bound conformation 667:696 the agonist-bound conformation of GBR1 667:704 Here we demonstrate that the GBR2 ectodomain directly interacts with the GBR1 ectodomain to increase agonist affinity by selectively stabilizing the agonist-bound conformation of GBR1. 22660477 0 51 theme human 68:72 arg1 GBR2 91:94 human GABA(B) receptor GBR2 68:94 human GABA(B) receptor GBR2 68:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 22660477 4 52 theme heterodimeric 379:391 arg1 assembly 393:400 a heterodimeric assembly 377:400 a heterodimeric assembly 377:400 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 4 52 theme heterodimeric 379:391 arg1 receptor 355:362 GABA(B) receptor 347:362 GABA(B) receptor 347:362 GABA(B) receptor functions as a heterodimeric assembly of GBR1 and GBR2 subunits, where GBR1 is responsible for ligand-binding and GBR2 is responsible for G protein coupling. 22660477 2 53 theme central 223:229 arg1 receptor 214:221 a G protein-coupled receptor 194:221 a G protein-coupled receptor central to mammalian brain function 194:257 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 2 53 theme central 223:229 arg1 receptor 182:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor 157:189 The metabotropic GABA(B) receptor is a G protein-coupled receptor central to mammalian brain function. 22660477 0 54 theme B 79:79 arg1 GBR2 91:94 human GABA(B) receptor GBR2 68:94 human GABA(B) receptor GBR2 68:94 Structure and functional interaction of the extracellular domain of human GABA(B) receptor GBR2. 23129617 4 0 theme surface 545:551 arg1 expression 553:562 surface expression 545:562 surface expression of AMPARs 545:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 6 1 theme endogenous 755:764 arg1 RNF167 766:771 endogenous RNF167 755:771 endogenous RNF167 755:771 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 8 2 theme excitatory 1236:1245 arg1 synapses 1247:1254 excitatory synapses 1236:1254 excitatory synapses 1236:1254 Therefore, our study identifies RNF167 as a selective regulator of AMPAR-mediated neurotransmission and expands our understanding of how ubiquitination dynamically regulates excitatory synapses. 23129617 4 3 theme AMPARs 567:572 arg1 expression 553:562 surface expression 545:562 surface expression of AMPARs 545:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 8 4 theme AMPAR-mediated 1129:1142 arg1 neurotransmission 1144:1160 AMPAR-mediated neurotransmission 1129:1160 AMPAR-mediated neurotransmission 1129:1160 Therefore, our study identifies RNF167 as a selective regulator of AMPAR-mediated neurotransmission and expands our understanding of how ubiquitination dynamically regulates excitatory synapses. 23129617 6 5 theme RNF167 867:872 arg1 activity 874:881 disrupted RNF167 activity 857:881 disrupted RNF167 activity 857:881 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 6 6 theme activity-dependent 914:931 arg1 ubiquitination 933:946 activity-dependent ubiquitination 914:946 activity-dependent ubiquitination of AMPARs 914:956 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 3 7 theme enzymes 393:399 arg1 few 367:369 few 367:369 few 367:369 However, very few of the ubiquitinating enzymes implicated in the process have been identified. 23129617 6 8 theme disrupted 857:865 arg1 activity 874:881 disrupted RNF167 activity 857:881 disrupted RNF167 activity 857:881 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 1 9 theme AMPA 80:83 arg1 receptors 85:93 AMPA receptors 80:93 AMPA receptors (AMPARs) 80:102 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 1 9 theme AMPA 80:83 arg1 AMPARs 96:101 AMPARs 96:101 AMPARs 96:101 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 6 10 theme specific 727:734 arg1 shRNA 736:740 a specific shRNA 725:740 a specific shRNA to eliminate endogenous RNF167 725:771 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 4 11 theme ubiquitin 513:521 arg1 ligases 523:529 transmembrane RING domain-containing E3 ubiquitin ligases 473:529 transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs 473:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 2 12 theme synaptic 315:322 arg1 expression 324:333 the synaptic expression 311:333 the synaptic expression of many proteins 311:350 Ubiquitination is a posttranslational modification that dynamically regulates the synaptic expression of many proteins. 23129617 3 13 theme ubiquitinating 378:391 arg1 enzymes 393:399 the ubiquitinating enzymes 374:399 the ubiquitinating enzymes implicated in the process 374:425 However, very few of the ubiquitinating enzymes implicated in the process have been identified. 23129617 8 14 theme selective 1106:1114 arg1 regulator 1116:1124 a selective regulator 1104:1124 a selective regulator of AMPAR-mediated neurotransmission 1104:1160 Therefore, our study identifies RNF167 as a selective regulator of AMPAR-mediated neurotransmission and expands our understanding of how ubiquitination dynamically regulates excitatory synapses. 23129617 8 14 theme selective 1106:1114 arg1 RNF167 1094:1099 RNF167 1094:1099 RNF167 1094:1099 Therefore, our study identifies RNF167 as a selective regulator of AMPAR-mediated neurotransmission and expands our understanding of how ubiquitination dynamically regulates excitatory synapses. 23129617 6 15 theme hippocampal 832:842 arg1 neurons 844:850 hippocampal neurons 832:850 hippocampal neurons with disrupted RNF167 activity 832:881 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 6 16 with neurons 844:850 arg1 activity 874:881 disrupted RNF167 activity 857:881 disrupted RNF167 activity 857:881 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 5 17 located located 651:657 arg1 surface 666:672 the surface 662:672 the surface of cultured neurons 662:692 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 17 located located 651:657 arg2 lysosomal 611:619 lysosomal 611:619 lysosomal 611:619 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 17 located located 651:657 arg2 located 651:657 located 651:657 located 651:657 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 17 located located 651:657 arg2 subpopulation 624:636 a subpopulation 622:636 a subpopulation of RNF167 622:646 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 18 theme cultured 677:684 arg1 neurons 686:692 cultured neurons 677:692 cultured neurons 677:692 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 6 19 theme AMPARs 951:956 arg1 ubiquitination 933:946 activity-dependent ubiquitination 914:946 activity-dependent ubiquitination of AMPARs 914:956 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 1 20 theme postsynaptic 184:195 arg1 sites 197:201 postsynaptic sites 184:201 postsynaptic sites 184:201 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 0 21 theme Ubiquitin 0:8 arg1 ligase 10:15 Ubiquitin ligase RNF167 0:22 Ubiquitin ligase RNF167 0:22 Ubiquitin ligase RNF167 regulates AMPA receptor-mediated synaptic transmission. 23129617 5 22 theme neurons 686:692 arg1 surface 666:672 the surface 662:672 the surface of cultured neurons 662:692 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 23 theme RNF167 641:646 arg1 located 651:657 located 651:657 located 651:657 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 23 theme RNF167 641:646 arg1 lysosomal 611:619 lysosomal 611:619 lysosomal 611:619 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 23 theme RNF167 641:646 arg1 subpopulation 624:636 a subpopulation 622:636 a subpopulation of RNF167 622:646 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 2 24 theme proteins 343:350 arg1 expression 324:333 the synaptic expression 311:333 the synaptic expression of many proteins 311:350 Ubiquitination is a posttranslational modification that dynamically regulates the synaptic expression of many proteins. 23129617 7 25 theme synaptic 1022:1029 arg1 currents 1037:1044 synaptic NMDAR currents 1022:1044 synaptic NMDAR currents 1022:1044 In addition, RNF167 regulates synaptic AMPAR currents, whereas synaptic NMDAR currents are unaffected. 23129617 8 26 theme neurotransmission 1144:1160 arg1 regulator 1116:1124 a selective regulator 1104:1124 a selective regulator of AMPAR-mediated neurotransmission 1104:1160 Therefore, our study identifies RNF167 as a selective regulator of AMPAR-mediated neurotransmission and expands our understanding of how ubiquitination dynamically regulates excitatory synapses. 23129617 8 26 theme neurotransmission 1144:1160 arg1 RNF167 1094:1099 RNF167 1094:1099 RNF167 1094:1099 Therefore, our study identifies RNF167 as a selective regulator of AMPAR-mediated neurotransmission and expands our understanding of how ubiquitination dynamically regulates excitatory synapses. 23129617 4 27 theme RING 487:490 arg1 ligases 523:529 transmembrane RING domain-containing E3 ubiquitin ligases 473:529 transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs 473:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 2 28 theme many 338:341 arg1 proteins 343:350 many proteins 338:350 many proteins 338:350 Ubiquitination is a posttranslational modification that dynamically regulates the synaptic expression of many proteins. 23129617 1 29 theme synaptic 214:221 arg1 strength 223:230 synaptic strength 214:230 synaptic strength 214:230 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 0 30 theme receptor-mediated 39:55 arg1 transmission 66:77 AMPA receptor-mediated synaptic transmission 34:77 AMPA receptor-mediated synaptic transmission 34:77 Ubiquitin ligase RNF167 regulates AMPA receptor-mediated synaptic transmission. 23129617 4 31 theme transmembrane 473:485 arg1 ligases 523:529 transmembrane RING domain-containing E3 ubiquitin ligases 473:529 transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs 473:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 2 32 theme posttranslational 253:269 arg1 Ubiquitination 233:246 Ubiquitination 233:246 Ubiquitination 233:246 Ubiquitination is a posttranslational modification that dynamically regulates the synaptic expression of many proteins. 23129617 2 32 theme posttranslational 253:269 arg1 modification 271:282 a posttranslational modification 251:282 a posttranslational modification that dynamically regulates the synaptic expression of many proteins 251:350 Ubiquitination is a posttranslational modification that dynamically regulates the synaptic expression of many proteins. 23129617 0 33 theme AMPA 34:37 arg1 transmission 66:77 AMPA receptor-mediated synaptic transmission 34:77 AMPA receptor-mediated synaptic transmission 34:77 Ubiquitin ligase RNF167 regulates AMPA receptor-mediated synaptic transmission. 23129617 4 34 theme E3 510:511 arg1 ligases 523:529 transmembrane RING domain-containing E3 ubiquitin ligases 473:529 transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs 473:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 6 35 theme surface 800:806 arg1 expression 808:817 AMPAR surface expression 794:817 AMPAR surface expression 794:817 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 4 36 theme domain-containing 492:508 arg1 ligases 523:529 transmembrane RING domain-containing E3 ubiquitin ligases 473:529 transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs 473:572 In a screen to identify transmembrane RING domain-containing E3 ubiquitin ligases that regulate surface expression of AMPARs, we identified RNF167. 23129617 6 37 theme AMPAR 794:798 arg1 expression 808:817 AMPAR surface expression 794:817 AMPAR surface expression 794:817 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 0 38 theme synaptic 57:64 arg1 transmission 66:77 AMPA receptor-mediated synaptic transmission 34:77 AMPA receptor-mediated synaptic transmission 34:77 Ubiquitin ligase RNF167 regulates AMPA receptor-mediated synaptic transmission. 23129617 6 39 theme RING 703:706 arg1 mutant 708:713 a RING mutant RNF167 701:720 a RING mutant RNF167 701:720 Using a RING mutant RNF167 or a specific shRNA to eliminate endogenous RNF167, we demonstrate that AMPAR surface expression increases in hippocampal neurons with disrupted RNF167 activity and that RNF167 is involved in activity-dependent ubiquitination of AMPARs. 23129617 7 40 theme synaptic 989:996 arg1 currents 1004:1011 synaptic AMPAR currents 989:1011 synaptic AMPAR currents 989:1011 In addition, RNF167 regulates synaptic AMPAR currents, whereas synaptic NMDAR currents are unaffected. 23129617 7 41 theme AMPAR 998:1002 arg1 currents 1004:1011 synaptic AMPAR currents 989:1011 synaptic AMPAR currents 989:1011 In addition, RNF167 regulates synaptic AMPAR currents, whereas synaptic NMDAR currents are unaffected. 23129617 7 42 theme NMDAR 1031:1035 arg1 currents 1037:1044 synaptic NMDAR currents 1022:1044 synaptic NMDAR currents 1022:1044 In addition, RNF167 regulates synaptic AMPAR currents, whereas synaptic NMDAR currents are unaffected. 23129617 1 43 theme fast 128:131 arg1 neurotransmission 144:160 fast excitatory neurotransmission 128:160 fast excitatory neurotransmission 128:160 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 1 44 theme excitatory 133:142 arg1 neurotransmission 144:160 fast excitatory neurotransmission 128:160 fast excitatory neurotransmission 128:160 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 1 45 from sites 197:201 arg1 density 173:179 their density 167:179 their density at postsynaptic sites 167:201 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 1 46 theme neurotransmission 144:160 arg1 majority 116:123 the majority 112:123 the majority of fast excitatory neurotransmission 112:160 AMPA receptors (AMPARs) mediate the majority of fast excitatory neurotransmission, and their density at postsynaptic sites determines synaptic strength. 23129617 5 47 dep located 651:657 arg1 located 651:657 located 651:657 located 651:657 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 47 dep located 651:657 arg1 lysosomal 611:619 lysosomal 611:619 lysosomal 611:619 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 5 47 dep located 651:657 arg1 subpopulation 624:636 a subpopulation 622:636 a subpopulation of RNF167 622:646 Predominantly lysosomal, a subpopulation of RNF167 is located on the surface of cultured neurons. 23129617 7 48 dep regulates 979:987 arg1 whereas 1014:1020 whereas 1014:1020 whereas 1014:1020 In addition, RNF167 regulates synaptic AMPAR currents, whereas synaptic NMDAR currents are unaffected. 21805521 5 0 link N-linked 864:871 arg1 glycans 873:879 three N-linked glycans 858:879 three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%) 858:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 4 1 link O-linked 678:685 arg1 glycans 687:693 O-linked glycans 678:693 O-linked glycans 678:693 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 6 2 theme GalNAc-Gal[sialic 1111:1127 arg1 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 2 theme GalNAc-Gal[sialic 1111:1127 arg1 % 1138:1138 30% 1136:1138 30% 1136:1138 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 2 theme GalNAc-Gal[sialic 1111:1127 arg1 acid 1129:1132 GalNAc-Gal[sialic acid] 1111:1133 GalNAc-Gal[sialic acid] (30%) 1111:1139 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 3 theme O-linked 1046:1053 arg1 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 3 theme O-linked 1046:1053 arg1 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 3 theme O-linked 1046:1053 arg1 acid 1129:1132 GalNAc-Gal[sialic acid] 1111:1133 GalNAc-Gal[sialic acid] (30%) 1111:1139 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 9 4 theme C-terminal 1764:1773 arg1 domain 1775:1780 the C-terminal domain 1760:1780 the C-terminal domain 1760:1780 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 3 5 theme K 523:523 arg1 weights 506:512 molecular weights 496:512 molecular weights of 27-28 K 496:523 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 3 5 theme K 523:523 arg1 K 532:532 30 K 529:532 30 K at ∼ 1:9 ratio 529:547 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 1 6 theme bone 250:253 arg1 metabolism 255:264 bone metabolism 250:264 bone metabolism 250:264 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 5 7 dep forms 909:913 arg1 % 951:951 60% 949:951 60% 949:951 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 7 dep forms 909:913 arg1 % 943:943 23% 941:943 23% 941:943 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 7 8 theme disulfide 1235:1243 arg1 bonds 1245:1249 The 10 intramolecular disulfide bonds 1213:1249 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains 1213:1296 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 4 9 theme 27.4K 606:610 arg1 weights 595:601 the observed molecular weights 572:601 the observed molecular weights of 27.4K and 29.5K 572:620 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 5 10 theme ∼ 811:811 arg1 huDKK1 820:825 huDKK1 820:825 huDKK1 820:825 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 10 theme ∼ 811:811 arg1 % 815:815 ∼ 92% 811:815 ∼ 92% of huDKK1 811:825 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 7 11 from N- 1258:1259 arg1 bonds 1245:1249 The 10 intramolecular disulfide bonds 1213:1249 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains 1213:1296 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 9 12 theme expected 1791:1798 arg1 homology 1800:1807 the expected homology 1787:1807 the expected homology with those found in colipase and other reported disulfide clusters 1787:1874 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 4 13 theme 29.5K 616:620 arg1 weights 595:601 the observed molecular weights 572:601 the observed molecular weights of 27.4K and 29.5K 572:620 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 8 14 theme family 1605:1610 arg1 proteins 1612:1619 the DKK family proteins 1597:1619 the DKK family proteins 1597:1619 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 4 15 theme observed 576:583 arg1 weights 595:601 the observed molecular weights 572:601 the observed molecular weights of 27.4K and 29.5K 572:620 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 4 16 contain contain 670:676 arg1 form 661:664 the low molecular weight form 636:664 the low molecular weight form 636:664 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 4 16 contain contain 670:676 arg2 glycans 687:693 O-linked glycans 678:693 O-linked glycans 678:693 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 5 17 theme peptide 780:786 arg1 mapping 788:794 LC-MS/MS peptide mapping 771:794 LC-MS/MS peptide mapping 771:794 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 7 18 from domains 1290:1296 arg1 bonds 1245:1249 The 10 intramolecular disulfide bonds 1213:1249 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains 1213:1296 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 19 theme peptides 1512:1519 arg1 characterization 1383:1398 characterization 1383:1398 characterization 1383:1398 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 19 theme peptides 1512:1519 arg1 isolation 1369:1377 isolation 1369:1377 isolation 1369:1377 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 19 theme peptides 1512:1519 arg1 characterization 1462:1477 characterization 1462:1477 characterization 1462:1477 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 19 theme peptides 1512:1519 arg1 digestions 1357:1366 multiple proteolytic digestions 1336:1366 multiple proteolytic digestions 1336:1366 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 19 theme peptides 1512:1519 arg1 digestion 1448:1456 secondary digestion 1438:1456 secondary digestion 1438:1456 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 6 20 theme -Gal-sialic 1084:1094 arg1 acid 1079:1082 sialic acid 1072:1082 sialic acid 1072:1082 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 20 theme -Gal-sialic 1084:1094 arg1 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 20 theme -Gal-sialic 1084:1094 arg1 % 1104:1104 65% 1102:1104 65% 1102:1104 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 20 theme -Gal-sialic 1084:1094 arg1 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 0 21 theme cysteine-rich 120:132 arg1 domains 134:140 the two cysteine-rich domains 112:140 the two cysteine-rich domains 112:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 1 22 theme canonical 190:198 arg1 pathway 214:220 the canonical Wnt-signaling pathway 186:220 the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases 186:283 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 7 23 theme selected 1482:1489 arg1 peptides 1512:1519 selected disulfide-containing peptides 1482:1519 selected disulfide-containing peptides 1482:1519 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 4 24 theme weight 654:659 arg1 form 661:664 the low molecular weight form 636:664 the low molecular weight form 636:664 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 3 25 theme 1:9 539:541 arg1 ratio 543:547 ∼ 1:9 ratio 537:547 ∼ 1:9 ratio 537:547 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 8 26 theme exact 1635:1639 arg1 matches 1641:1647 no exact matches 1632:1647 no exact matches in disulfide positioning 1632:1672 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 5 27 gly glycosylated 830:841 arg2 Asn²²⁵ 846:851 Asn²²⁵ 846:851 Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%) 846:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 27 gly glycosylated 830:841 arg1 huDKK1 820:825 huDKK1 820:825 huDKK1 820:825 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 27 gly glycosylated 830:841 arg1 % 815:815 ∼ 92% 811:815 ∼ 92% of huDKK1 811:825 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 0 28 from linkages 100:107 arg1 domains 134:140 the two cysteine-rich domains 112:140 the two cysteine-rich domains 112:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 9 29 theme other 1842:1846 arg1 clusters 1867:1874 other reported disulfide clusters 1842:1874 other reported disulfide clusters 1842:1874 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 9 30 theme disulfide 1857:1865 arg1 clusters 1867:1874 other reported disulfide clusters 1842:1874 other reported disulfide clusters 1842:1874 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 6 31 link O-linked 1046:1053 arg1 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 31 link O-linked 1046:1053 arg1 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 31 link O-linked 1046:1053 arg1 acid 1129:1132 GalNAc-Gal[sialic acid] 1111:1133 GalNAc-Gal[sialic acid] (30%) 1111:1139 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 32 theme targeted 1194:1201 arg1 LC-MS/MS 1203:1210 β-elimination and targeted LC-MS/MS 1176:1210 β-elimination and targeted LC-MS/MS 1176:1210 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 1 33 theme engineered 303:312 arg1 cells 336:340 engineered Chinese hamster ovary cells 303:340 engineered Chinese hamster ovary cells 303:340 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 1 34 theme Human 143:147 arg1 huDKK1 161:166 huDKK1 161:166 huDKK1 161:166 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 1 34 theme Human 143:147 arg1 Dickkopf-1 149:158 Human Dickkopf-1 143:158 Human Dickkopf-1 (huDKK1) 143:167 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 1 34 theme Human 143:147 arg1 inhibitor 173:181 an inhibitor 170:181 an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases 170:283 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 4 35 theme low 640:642 arg1 weight 654:659 the low molecular weight 636:659 the low molecular weight form 636:664 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 6 36 theme β-elimination 1176:1188 arg1 LC-MS/MS 1203:1210 β-elimination and targeted LC-MS/MS 1176:1210 β-elimination and targeted LC-MS/MS 1176:1210 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 1 37 theme hamster 322:328 arg1 cells 336:340 engineered Chinese hamster ovary cells 303:340 engineered Chinese hamster ovary cells 303:340 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 7 38 theme peptides 1424:1431 arg1 characterization 1383:1398 characterization 1383:1398 characterization 1383:1398 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 38 theme peptides 1424:1431 arg1 isolation 1369:1377 isolation 1369:1377 isolation 1369:1377 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 38 theme peptides 1424:1431 arg1 characterization 1462:1477 characterization 1462:1477 characterization 1462:1477 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 38 theme peptides 1424:1431 arg1 digestions 1357:1366 multiple proteolytic digestions 1336:1366 multiple proteolytic digestions 1336:1366 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 38 theme peptides 1424:1431 arg1 digestion 1448:1456 secondary digestion 1438:1456 secondary digestion 1438:1456 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 4 39 theme high 705:708 arg1 weight 720:725 the high molecular weight 701:725 the high molecular weight form 701:730 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 8 40 theme other 1691:1695 arg1 clusters 1713:1720 other known disulfide clusters 1691:1720 other known disulfide clusters 1691:1720 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 5 41 theme biantennary 897:907 arg1 forms 909:913 two biantennary forms 893:913 two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively) 893:966 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 4 42 theme weight 720:725 arg1 form 727:730 the high molecular weight form 701:730 the high molecular weight form 701:730 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 0 43 theme linkages 100:107 arg1 determination 73:85 determination 73:85 determination of disulfide linkages in the two cysteine-rich domains 73:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 0 43 theme linkages 100:107 arg1 characterization 35:50 characterization 35:50 characterization of glycosylation 35:67 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 9 44 located found 1820:1824 arg2 those 1814:1818 those 1814:1818 those 1814:1818 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 9 44 located found 1820:1824 arg1 clusters 1867:1874 other reported disulfide clusters 1842:1874 other reported disulfide clusters 1842:1874 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 9 44 located found 1820:1824 arg1 colipase 1829:1836 colipase 1829:1836 colipase 1829:1836 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 8 45 theme disulfide 1703:1711 arg1 clusters 1713:1720 other known disulfide clusters 1691:1720 other known disulfide clusters 1691:1720 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 4 46 theme O-linked 678:685 arg1 glycans 687:693 O-linked glycans 678:693 O-linked glycans 678:693 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 7 47 theme cysteine-rich 1276:1288 arg1 domains 1290:1296 C-terminal cysteine-rich domains 1265:1296 C-terminal cysteine-rich domains 1265:1296 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 0 48 theme Human 0:4 arg1 protein 26:32 Human Dickkopf-1 (huDKK1) protein 0:32 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 0:141 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 2 49 theme reporter 410:417 arg1 assay 424:428 a TCF/lef-luciferase reporter gene assay 389:428 a TCF/lef-luciferase reporter gene assay 389:428 HuDKK1 is biologically active in a TCF/lef-luciferase reporter gene assay and is able to bind LRP6 coreceptor. 21805521 1 50 theme other 270:274 arg1 diseases 276:283 other diseases 270:283 other diseases 270:283 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 2 51 theme LRP6 450:453 arg1 coreceptor 455:464 LRP6 coreceptor 450:464 LRP6 coreceptor 450:464 HuDKK1 is biologically active in a TCF/lef-luciferase reporter gene assay and is able to bind LRP6 coreceptor. 21805521 4 52 theme O-linked 753:760 arg1 glycans 762:768 O-linked glycans 753:768 O-linked glycans 753:768 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 7 53 dep N- 1258:1259 arg1 the 1254:1256 the 1254:1256 the 1254:1256 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 54 theme multiple 1336:1343 arg1 digestions 1357:1366 multiple proteolytic digestions 1336:1366 multiple proteolytic digestions 1336:1366 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 2 55 from active 379:384 arg1 assay 424:428 a TCF/lef-luciferase reporter gene assay 389:428 a TCF/lef-luciferase reporter gene assay 389:428 HuDKK1 is biologically active in a TCF/lef-luciferase reporter gene assay and is able to bind LRP6 coreceptor. 21805521 1 56 theme Wnt-signaling 200:212 arg1 pathway 214:220 the canonical Wnt-signaling pathway 186:220 the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases 186:283 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 5 57 theme huDKK1 820:825 arg1 huDKK1 820:825 huDKK1 820:825 huDKK1 820:825 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 57 theme huDKK1 820:825 arg1 % 815:815 ∼ 92% 811:815 ∼ 92% of huDKK1 811:825 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 9 58 theme disulfide 1732:1740 arg1 bonds 1742:1746 The five disulfide bonds 1723:1746 The five disulfide bonds assigned in the C-terminal domain 1723:1780 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 6 59 contain contains 1033:1040 arg2 acid 1129:1132 GalNAc-Gal[sialic acid] 1111:1133 GalNAc-Gal[sialic acid] (30%) 1111:1139 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 59 contain contains 1033:1040 arg2 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 59 contain contains 1033:1040 arg1 HuDKK1 1026:1031 HuDKK1 1026:1031 HuDKK1 1026:1031 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 59 contain contains 1033:1040 arg2 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 5 60 theme sialic 928:933 arg1 acid 935:938 1 and 2 sialic acid 920:938 1 and 2 sialic acid 920:938 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 6 61 attach attached 1142:1149 arg3 Ser³⁰ 1154:1158 Ser³⁰ 1154:1158 Ser³⁰ 1154:1158 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 61 attach attached 1142:1149 arg2 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 61 attach attached 1142:1149 arg2 acid 1129:1132 GalNAc-Gal[sialic acid] 1111:1133 GalNAc-Gal[sialic acid] (30%) 1111:1139 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 61 attach attached 1142:1149 arg2 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 5 62 theme sialic 1007:1012 arg1 acids 1014:1018 2 sialic acids 1005:1018 2 sialic acids (9%) 1005:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 62 theme sialic 1007:1012 arg1 % 1022:1022 9% 1021:1022 9% 1021:1022 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 3 63 theme 27-28 517:521 arg1 K 523:523 27-28 K 517:523 27-28 K 517:523 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 8 64 theme N-terminal 1565:1574 arg1 domain 1576:1581 the huDKK1 N-terminal domain 1554:1581 the huDKK1 N-terminal domain 1554:1581 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 7 65 theme C-terminal 1265:1274 arg1 domains 1290:1296 C-terminal cysteine-rich domains 1265:1296 C-terminal cysteine-rich domains 1265:1296 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 7 66 theme intramolecular 1220:1233 arg1 bonds 1245:1249 The 10 intramolecular disulfide bonds 1213:1249 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains 1213:1296 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 8 67 theme DKK 1601:1603 arg1 proteins 1612:1619 the DKK family proteins 1597:1619 the DKK family proteins 1597:1619 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 5 68 theme LC-MS/MS 771:778 arg1 mapping 788:794 LC-MS/MS peptide mapping 771:794 LC-MS/MS peptide mapping 771:794 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 7 69 theme disulfide-containing 1491:1510 arg1 peptides 1512:1519 selected disulfide-containing peptides 1482:1519 selected disulfide-containing peptides 1482:1519 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 0 70 from determination 73:85 arg1 domains 134:140 the two cysteine-rich domains 112:140 the two cysteine-rich domains 112:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 5 71 with forms 909:913 arg1 acids 1014:1018 2 sialic acids 1005:1018 2 sialic acids (9%) 1005:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 71 with forms 909:913 arg1 % 1022:1022 9% 1021:1022 9% 1021:1022 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 71 with forms 909:913 arg1 acid 935:938 1 and 2 sialic acid 920:938 1 and 2 sialic acid 920:938 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 4 72 theme molecular 585:593 arg1 weights 595:601 the observed molecular weights 572:601 the observed molecular weights of 27.4K and 29.5K 572:620 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 3 73 theme ∼ 537:537 arg1 ratio 543:547 ∼ 1:9 ratio 537:547 ∼ 1:9 ratio 537:547 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 6 74 theme sialic 1072:1077 arg1 acid 1079:1082 sialic acid 1072:1082 sialic acid 1072:1082 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 74 theme sialic 1072:1077 arg1 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 4 75 theme molecular 644:652 arg1 weight 654:659 the low molecular weight 636:659 the low molecular weight form 636:664 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 5 76 with structure 990:998 arg1 acids 1014:1018 2 sialic acids 1005:1018 2 sialic acids (9%) 1005:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 76 with structure 990:998 arg1 % 1022:1022 9% 1021:1022 9% 1021:1022 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 5 76 with structure 990:998 arg1 acid 935:938 1 and 2 sialic acid 920:938 1 and 2 sialic acid 920:938 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 7 77 theme secondary 1438:1446 arg1 digestion 1448:1456 secondary digestion 1438:1456 secondary digestion 1438:1456 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 6 78 theme GalNAc 1064:1069 arg1 acid 1079:1082 sialic acid 1072:1082 sialic acid 1072:1082 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 78 theme GalNAc 1064:1069 arg1 acid 1096:1099 GalNAc (sialic acid)-Gal-sialic acid 1064:1099 GalNAc (sialic acid)-Gal-sialic acid (65%) 1064:1105 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 78 theme GalNAc 1064:1069 arg1 % 1104:1104 65% 1102:1104 65% 1102:1104 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 6 78 theme GalNAc 1064:1069 arg1 glycans 1055:1061 two O-linked glycans 1042:1061 two O-linked glycans 1042:1061 HuDKK1 contains two O-linked glycans, GalNAc (sialic acid)-Gal-sialic acid (65%) and GalNAc-Gal[sialic acid] (30%), attached at Ser³⁰ as confirmed by β-elimination and targeted LC-MS/MS. 21805521 9 79 theme reported 1848:1855 arg1 clusters 1867:1874 other reported disulfide clusters 1842:1874 other reported disulfide clusters 1842:1874 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 0 80 theme glycosylation 55:67 arg1 determination 73:85 determination 73:85 determination of disulfide linkages in the two cysteine-rich domains 73:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 0 80 theme glycosylation 55:67 arg1 characterization 35:50 characterization 35:50 characterization of glycosylation 35:67 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 2 81 from assay 424:428 arg1 active 379:384 active 379:384 active 379:384 HuDKK1 is biologically active in a TCF/lef-luciferase reporter gene assay and is able to bind LRP6 coreceptor. 21805521 0 82 from domains 134:140 arg1 determination 73:85 determination 73:85 determination of disulfide linkages in the two cysteine-rich domains 73:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 0 82 from domains 134:140 arg1 characterization 35:50 characterization 35:50 characterization of glycosylation 35:67 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 4 83 contain contains 732:739 arg1 form 727:730 the high molecular weight form 701:730 the high molecular weight form 701:730 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 4 83 contain contains 732:739 arg2 glycans 762:768 O-linked glycans 753:768 O-linked glycans 753:768 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 4 83 contain contains 732:739 arg2 N- 746:747 N- 746:747 N- 746:747 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 1 84 theme Chinese 314:320 arg1 cells 336:340 engineered Chinese hamster ovary cells 303:340 engineered Chinese hamster ovary cells 303:340 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 8 85 theme disulfide 1652:1660 arg1 positioning 1662:1672 disulfide positioning 1652:1672 disulfide positioning 1652:1672 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 7 86 theme disulfide-containing 1403:1422 arg1 peptides 1424:1431 disulfide-containing peptides 1403:1431 disulfide-containing peptides 1403:1431 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 0 87 theme disulfide 90:98 arg1 linkages 100:107 disulfide linkages 90:107 disulfide linkages in the two cysteine-rich domains 90:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 1 88 theme ovary 330:334 arg1 cells 336:340 engineered Chinese hamster ovary cells 303:340 engineered Chinese hamster ovary cells 303:340 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 0 89 from characterization 35:50 arg1 domains 134:140 the two cysteine-rich domains 112:140 the two cysteine-rich domains 112:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 5 90 theme triantennary 977:988 arg1 structure 990:998 one triantennary structure 973:998 one triantennary structure with 2 sialic acids (9%) 973:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 4 91 theme molecular 710:718 arg1 weight 720:725 the high molecular weight 701:725 the high molecular weight form 701:730 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 0 92 theme Dickkopf-1 6:15 arg1 protein 26:32 Human Dickkopf-1 (huDKK1) protein 0:32 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 0:141 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 8 93 theme known 1697:1701 arg1 clusters 1713:1720 other known disulfide clusters 1691:1720 other known disulfide clusters 1691:1720 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 5 94 theme N-linked 864:871 arg1 glycans 873:879 three N-linked glycans 858:879 three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%) 858:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 4 95 link O-linked 753:760 arg1 glycans 762:768 O-linked glycans 753:768 O-linked glycans 753:768 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 5 96 with Asn²²⁵ 846:851 arg1 glycans 873:879 three N-linked glycans 858:879 three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%) 858:1023 LC-MS/MS peptide mapping indicates that ∼ 92% of huDKK1 is glycosylated at Asn²²⁵ with three N-linked glycans composed of two biantennary forms with 1 and 2 sialic acid (23% and 60%, respectively), and one triantennary structure with 2 sialic acids (9%). 21805521 0 97 theme huDKK1 18:23 arg1 protein 26:32 Human Dickkopf-1 (huDKK1) protein 0:32 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 0:141 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 2 98 theme gene 419:422 arg1 assay 424:428 a TCF/lef-luciferase reporter gene assay 389:428 a TCF/lef-luciferase reporter gene assay 389:428 HuDKK1 is biologically active in a TCF/lef-luciferase reporter gene assay and is able to bind LRP6 coreceptor. 21805521 3 99 from ratio 543:547 arg1 weights 506:512 molecular weights 496:512 molecular weights of 27-28 K 496:523 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 3 99 from ratio 543:547 arg1 K 532:532 30 K 529:532 30 K at ∼ 1:9 ratio 529:547 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 2 100 theme TCF/lef-luciferase 391:408 arg1 assay 424:428 a TCF/lef-luciferase reporter gene assay 389:428 a TCF/lef-luciferase reporter gene assay 389:428 HuDKK1 is biologically active in a TCF/lef-luciferase reporter gene assay and is able to bind LRP6 coreceptor. 21805521 9 101 with homology 1800:1807 arg1 those 1814:1818 those 1814:1818 those 1814:1818 The five disulfide bonds assigned in the C-terminal domain show the expected homology with those found in colipase and other reported disulfide clusters. 21805521 0 102 dep protein 26:32 arg1 determination 73:85 determination 73:85 determination of disulfide linkages in the two cysteine-rich domains 73:140 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 0 102 dep protein 26:32 arg1 characterization 35:50 characterization 35:50 characterization of glycosylation 35:67 Human Dickkopf-1 (huDKK1) protein: characterization of glycosylation and determination of disulfide linkages in the two cysteine-rich domains. 21805521 8 103 theme disulfide 1531:1539 arg1 bonds 1541:1545 The five disulfide bonds 1522:1545 The five disulfide bonds within the huDKK1 N-terminal domain 1522:1581 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 8 103 theme disulfide 1531:1539 arg1 unique 1587:1592 unique 1587:1592 unique 1587:1592 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 7 104 theme proteolytic 1345:1355 arg1 digestions 1357:1366 multiple proteolytic digestions 1336:1366 multiple proteolytic digestions 1336:1366 The 10 intramolecular disulfide bonds at the N- and C-terminal cysteine-rich domains were elucidated by analyses including multiple proteolytic digestions, isolation and characterization of disulfide-containing peptides, and secondary digestion and characterization of selected disulfide-containing peptides. 21805521 4 105 theme MALDI-MS 553:560 arg1 analysis 562:569 MALDI-MS analysis 553:569 MALDI-MS analysis 553:569 By MALDI-MS analysis, the observed molecular weights of 27.4K and 29.5K indicate that the low molecular weight form may contain O-linked glycans while the high molecular weight form contains both N- and O-linked glycans. 21805521 1 106 theme pathway 214:220 arg1 Dickkopf-1 149:158 Human Dickkopf-1 143:158 Human Dickkopf-1 (huDKK1) 143:167 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 1 106 theme pathway 214:220 arg1 inhibitor 173:181 an inhibitor 170:181 an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases 170:283 Human Dickkopf-1 (huDKK1), an inhibitor of the canonical Wnt-signaling pathway that has been implicated in bone metabolism and other diseases, was expressed in engineered Chinese hamster ovary cells and purified. 21805521 8 107 from matches 1641:1647 arg1 positioning 1662:1672 disulfide positioning 1652:1672 disulfide positioning 1652:1672 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 21805521 3 108 theme molecular 496:504 arg1 weights 506:512 molecular weights 496:512 molecular weights of 27-28 K 496:523 In SDS-PAGE, huDKK1 exhibits molecular weights of 27-28 K and 30 K at ∼ 1:9 ratio. 21805521 8 109 theme huDKK1 1558:1563 arg1 domain 1576:1581 the huDKK1 N-terminal domain 1554:1581 the huDKK1 N-terminal domain 1554:1581 The five disulfide bonds within the huDKK1 N-terminal domain are unique to the DKK family proteins; there are no exact matches in disulfide positioning when compared to other known disulfide clusters. 22327295 4 0 dep are 530:532 arg1 targeted 572:579 targeted 572:579 is specifically targeted 556:579 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 4 0 dep are 530:532 arg1 liberate 603:610 liberate 603:610 liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB 603:778 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 6 1 theme rational 943:950 arg1 basis 952:956 a rational basis 941:956 a rational basis for the specificity of TbpA for human transferrin 941:1006 Our studies provide a rational basis for the specificity of TbpA for human transferrin, show how TbpA promotes iron release from transferrin, and elucidate how TbpB facilitates this process. 22327295 4 2 theme corresponding 749:761 arg1 TbpB 775:778 the corresponding co-receptor TbpB 745:778 the corresponding co-receptor TbpB 745:778 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 1 3 theme human 81:85 arg1 pathogens 87:95 obligate human pathogens 72:95 obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea 72:152 Neisseria are obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea. 22327295 1 3 theme human 81:85 arg1 Neisseria 58:66 Neisseria 58:66 Neisseria 58:66 Neisseria are obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea. 22327295 3 4 theme membrane 340:347 arg1 protein 349:355 an integral outer membrane protein 322:355 an integral outer membrane protein 322:355 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 4 theme membrane 340:347 arg1 TbpA 316:319 TbpA 316:319 TbpA 316:319 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 4 5 theme TbpB 775:778 arg1 structures 695:704 crystal structures 687:704 crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB 687:778 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 5 6 theme TbpA-TbpB-transferrin 867:887 arg1 complex 889:895 the TbpA-TbpB-transferrin complex 863:895 the TbpA-TbpB-transferrin complex 863:895 We characterized the TbpB-transferrin complex by small-angle X-ray scattering and the TbpA-TbpB-transferrin complex by electron microscopy. 22327295 6 7 theme human 990:994 arg1 transferrin 996:1006 human transferrin 990:1006 human transferrin 990:1006 Our studies provide a rational basis for the specificity of TbpA for human transferrin, show how TbpA promotes iron release from transferrin, and elucidate how TbpB facilitates this process. 22327295 1 8 theme obligate 72:79 arg1 pathogens 87:95 obligate human pathogens 72:95 obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea 72:152 Neisseria are obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea. 22327295 1 8 theme obligate 72:79 arg1 Neisseria 58:66 Neisseria 58:66 Neisseria 58:66 Neisseria are obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea. 22327295 5 9 theme TbpB-transferrin 802:817 arg1 complex 819:825 the TbpB-transferrin complex 798:825 the TbpB-transferrin complex 798:825 We characterized the TbpB-transferrin complex by small-angle X-ray scattering and the TbpA-TbpB-transferrin complex by electron microscopy. 22327295 6 10 from transferrin 1050:1060 arg1 release 1037:1043 iron release 1032:1043 iron release from transferrin 1032:1060 Our studies provide a rational basis for the specificity of TbpA for human transferrin, show how TbpA promotes iron release from transferrin, and elucidate how TbpB facilitates this process. 22327295 4 11 theme complex 730:736 arg1 structures 695:704 crystal structures 687:704 crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB 687:778 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 2 12 from transferrin 230:240 arg1 extract 199:205 extract 199:205 extract 199:205 Neisseria require iron for survival and can extract it directly from human transferrin for transport across the outer membrane. 22327295 0 13 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for iron piracy by pathogenic Neisseria. 0:56 Structural basis for iron piracy by pathogenic Neisseria. 22327295 4 14 theme TbpA-transferrin 713:728 arg1 complex 730:736 the TbpA-transferrin complex 709:736 the TbpA-transferrin complex 709:736 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 5 15 theme electron 900:907 arg1 microscopy 909:918 electron microscopy 900:918 electron microscopy 900:918 We characterized the TbpB-transferrin complex by small-angle X-ray scattering and the TbpA-TbpB-transferrin complex by electron microscopy. 22327295 0 16 theme iron 21:24 arg1 piracy 26:31 iron piracy 21:31 iron piracy 21:31 Structural basis for iron piracy by pathogenic Neisseria. 22327295 4 17 theme human 538:542 arg1 transferrin 544:554 human transferrin 538:554 human transferrin 538:554 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 3 18 theme important 442:450 arg1 proteins 417:424 both proteins 412:424 both proteins 412:424 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 18 theme important 442:450 arg1 vaccine 452:458 potentially important vaccine and therapeutic targets 430:482 vaccine 452:458 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 5 19 theme small-angle 830:840 arg1 scattering 848:857 small-angle X-ray scattering 830:857 small-angle X-ray scattering 830:857 We characterized the TbpB-transferrin complex by small-angle X-ray scattering and the TbpA-TbpB-transferrin complex by electron microscopy. 22327295 3 20 theme transport 287:295 arg1 system 297:302 The transport system 283:302 The transport system 283:302 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 1 21 theme bacterial 105:113 arg1 meningitis 115:124 bacterial meningitis 105:124 bacterial meningitis 105:124 Neisseria are obligate human pathogens causing bacterial meningitis, septicaemia and gonorrhoea. 22327295 4 22 from transferrin 622:632 arg1 iron 612:615 iron 612:615 iron from transferrin 612:632 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 6 23 theme TbpA 981:984 arg1 specificity 966:976 the specificity 962:976 the specificity of TbpA for human transferrin 962:1006 Our studies provide a rational basis for the specificity of TbpA for human transferrin, show how TbpA promotes iron release from transferrin, and elucidate how TbpB facilitates this process. 22327295 3 24 theme cell 398:401 arg1 surface 403:409 the cell surface 394:409 the cell surface 394:409 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 4 25 theme Neisseria 511:519 arg1 research 521:528 Neisseria research 511:528 Neisseria research 511:528 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 3 26 attach attached 382:389 arg2 TbpB 362:365 TbpB 362:365 TbpB 362:365 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 26 attach attached 382:389 arg1 surface 403:409 the cell surface 394:409 the cell surface 394:409 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 26 attach attached 382:389 arg2 co-receptor 370:380 a co-receptor 368:380 a co-receptor attached to the cell surface 368:409 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 2 27 theme human 224:228 arg1 transferrin 230:240 human transferrin 224:240 human transferrin 224:240 Neisseria require iron for survival and can extract it directly from human transferrin for transport across the outer membrane. 22327295 0 28 theme pathogenic 36:45 arg1 Neisseria 47:55 pathogenic Neisseria 36:55 pathogenic Neisseria 36:55 Structural basis for iron piracy by pathogenic Neisseria. 22327295 4 29 theme crystal 687:693 arg1 structures 695:704 crystal structures 687:704 crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB 687:778 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 2 30 theme outer 267:271 arg1 membrane 273:280 the outer membrane 263:280 the outer membrane 263:280 Neisseria require iron for survival and can extract it directly from human transferrin for transport across the outer membrane. 22327295 4 31 theme co-receptor 763:773 arg1 TbpB 775:778 the corresponding co-receptor TbpB 745:778 the corresponding co-receptor TbpB 745:778 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 3 32 theme therapeutic 464:474 arg1 targets 476:482 potentially important vaccine and therapeutic targets 430:482 targets 476:482 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 33 theme integral 325:332 arg1 protein 349:355 an integral outer membrane protein 322:355 an integral outer membrane protein 322:355 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 33 theme integral 325:332 arg1 TbpA 316:319 TbpA 316:319 TbpA 316:319 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 5 34 theme X-ray 842:846 arg1 scattering 848:857 small-angle X-ray scattering 830:857 small-angle X-ray scattering 830:857 We characterized the TbpB-transferrin complex by small-angle X-ray scattering and the TbpA-TbpB-transferrin complex by electron microscopy. 22327295 6 35 theme iron 1032:1035 arg1 release 1037:1043 iron release 1032:1043 iron release from transferrin 1032:1060 Our studies provide a rational basis for the specificity of TbpA for human transferrin, show how TbpA promotes iron release from transferrin, and elucidate how TbpB facilitates this process. 22327295 3 36 theme outer 334:338 arg1 protein 349:355 an integral outer membrane protein 322:355 an integral outer membrane protein 322:355 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 3 36 theme outer 334:338 arg1 TbpA 316:319 TbpA 316:319 TbpA 316:319 The transport system consists of TbpA, an integral outer membrane protein, and TbpB, a co-receptor attached to the cell surface; both proteins are potentially important vaccine and therapeutic targets. 22327295 4 37 theme key 489:491 arg1 questions 493:501 Two key questions 485:501 Two key questions driving Neisseria research 485:528 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 22327295 4 38 dep pH. 645:647 arg1 address 652:658 address 652:658 To address these questions 649:674 Two key questions driving Neisseria research are how human transferrin is specifically targeted, and how the bacteria liberate iron from transferrin at neutral pH. To address these questions, we solved crystal structures of the TbpA-transferrin complex and of the corresponding co-receptor TbpB. 15861141 2 0 theme interaction 465:475 arg1 inhibitor 400:408 a competitive peptide inhibitor 378:408 a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction 378:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 4 1 theme human 851:855 arg1 complex 866:872 the human uPA-uPAR complex 847:872 the human uPA-uPAR complex 847:872 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 15861141 1 2 theme soluble 141:147 arg1 form 149:152 a soluble form 139:152 a soluble form 139:152 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 6 3 with interplay 1147:1155 arg1 partners 1166:1173 the partners 1162:1173 the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration 1162:1286 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 2 4 theme -uPAR 459:463 arg1 interaction 465:475 the urokinase-type plasminogen activator (uPA)-uPAR interaction 413:475 the urokinase-type plasminogen activator (uPA)-uPAR interaction 413:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 6 5 theme cell 1235:1238 arg1 surface 1240:1246 the cell surface 1231:1246 the cell surface 1231:1246 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 1 6 theme form 149:152 arg1 structure 126:134 the crystal structure 114:134 the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers 114:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 2 7 with association 361:371 arg1 inhibitor 400:408 a competitive peptide inhibitor 378:408 a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction 378:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 1 8 from microenvironment 284:299 arg1 cancers 315:321 many human cancers 304:321 many human cancers 304:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 5 9 theme uPAR 946:949 arg1 cavity 959:964 the uPAR central cavity 942:964 the uPAR central cavity 942:964 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 0 10 theme antagonist 84:93 arg1 peptide 95:101 an antagonist peptide 81:101 an antagonist peptide 81:101 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 1 11 theme invasive 248:255 arg1 areas 257:261 the invasive areas 244:261 the invasive areas of the tumor-stromal microenvironment in many human cancers 244:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 1 12 from cancers 315:321 arg1 areas 257:261 the invasive areas 244:261 the invasive areas of the tumor-stromal microenvironment in many human cancers 244:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 2 13 theme peptide 392:398 arg1 inhibitor 400:408 a competitive peptide inhibitor 378:408 a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction 378:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 3 14 theme consecutive 504:514 arg1 domains 529:535 three consecutive three-finger domains 498:535 three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, 498:670 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 4 15 theme convergent 750:759 arg1 motif 769:773 a convergent binding motif 748:773 a convergent binding motif shared by the antagonist peptide and uPA 748:814 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 15861141 2 16 theme activator 444:452 arg1 interaction 465:475 the urokinase-type plasminogen activator (uPA)-uPAR interaction 413:475 the urokinase-type plasminogen activator (uPA)-uPAR interaction 413:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 3 17 theme large 678:682 arg1 surface 693:699 a large external surface 676:699 a large external surface 676:699 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 6 18 theme uPA-focalized 1202:1214 arg1 proteolysis 1216:1226 uPA-focalized proteolysis 1202:1226 uPA-focalized proteolysis 1202:1226 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 2 19 theme urokinase-type 417:430 arg1 uPA 455:457 uPA 455:457 uPA 455:457 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 2 19 theme urokinase-type 417:430 arg1 activator 444:452 the urokinase-type plasminogen activator 413:452 the urokinase-type plasminogen activator (uPA)-uPAR interaction 413:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 1 20 theme human 157:161 arg1 receptor 200:207 human urokinase-type plasminogen activator receptor 157:207 human urokinase-type plasminogen activator receptor (uPAR/CD87) 157:219 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 1 20 theme human 157:161 arg1 uPAR/CD87 210:218 uPAR/CD87 210:218 uPAR/CD87 210:218 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 2 21 theme plasminogen 432:442 arg1 uPA 455:457 uPA 455:457 uPA 455:457 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 2 21 theme plasminogen 432:442 arg1 activator 444:452 the urokinase-type plasminogen activator 413:452 the urokinase-type plasminogen activator (uPA)-uPAR interaction 413:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 3 22 theme external 684:691 arg1 surface 693:699 a large external surface 676:699 a large external surface 676:699 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 1 23 from areas 257:261 arg1 cancers 315:321 many human cancers 304:321 many human cancers 304:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 6 24 theme cell 1260:1263 arg1 adhesion 1265:1272 cell adhesion 1260:1272 cell adhesion 1260:1272 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 1 25 theme urokinase-type 163:176 arg1 receptor 200:207 human urokinase-type plasminogen activator receptor 157:207 human urokinase-type plasminogen activator receptor (uPAR/CD87) 157:219 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 1 25 theme urokinase-type 163:176 arg1 uPAR/CD87 210:218 uPAR/CD87 210:218 uPAR/CD87 210:218 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 0 26 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human urokinase plasminogen activator receptor 0:70 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 5 27 theme accessible 1010:1019 arg1 surface 1002:1008 the external receptor surface 980:1008 the external receptor surface accessible for other protein interactions (vitronectin and integrins) 980:1078 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 3 28 theme circular 560:567 arg1 manner 569:574 an almost circular manner 550:574 an almost circular manner 550:574 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 1 29 theme plasminogen 178:188 arg1 receptor 200:207 human urokinase-type plasminogen activator receptor 157:207 human urokinase-type plasminogen activator receptor (uPAR/CD87) 157:219 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 1 29 theme plasminogen 178:188 arg1 uPAR/CD87 210:218 uPAR/CD87 210:218 uPAR/CD87 210:218 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 6 30 theme structural 1096:1105 arg1 assembly 1107:1114 this unique structural assembly 1084:1114 this unique structural assembly 1084:1114 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 1 31 theme activator 190:198 arg1 receptor 200:207 human urokinase-type plasminogen activator receptor 157:207 human urokinase-type plasminogen activator receptor (uPAR/CD87) 157:219 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 1 31 theme activator 190:198 arg1 uPAR/CD87 210:218 uPAR/CD87 210:218 uPAR/CD87 210:218 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 4 32 theme complex 866:872 arg1 model 838:842 a model 836:842 a model of the human uPA-uPAR complex 836:872 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 15861141 5 33 theme receptor-binding 903:918 arg1 module 920:925 the receptor-binding module 899:925 the receptor-binding module of uPA 899:932 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 1 34 theme receptor 200:207 arg1 form 149:152 a soluble form 139:152 a soluble form 139:152 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 0 35 theme urokinase 31:39 arg1 receptor 63:70 the human urokinase plasminogen activator receptor 21:70 the human urokinase plasminogen activator receptor 21:70 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 1 36 theme tumor-stromal 270:282 arg1 microenvironment 284:299 the tumor-stromal microenvironment 266:299 the tumor-stromal microenvironment in many human cancers 266:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 5 37 theme other 1025:1029 arg1 interactions 1039:1050 other protein interactions 1025:1050 other protein interactions (vitronectin and integrins) 1025:1078 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 6 38 theme fine 1142:1145 arg1 interplay 1147:1155 the fine interplay 1138:1155 the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration 1138:1286 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 2 39 theme competitive 380:390 arg1 inhibitor 400:408 a competitive peptide inhibitor 378:408 a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction 378:475 The structure was solved at 2.7 A in association with a competitive peptide inhibitor of the urokinase-type plasminogen activator (uPA)-uPAR interaction. 15861141 0 40 theme human 25:29 arg1 receptor 63:70 the human urokinase plasminogen activator receptor 21:70 the human urokinase plasminogen activator receptor 21:70 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 1 41 theme microenvironment 284:299 arg1 areas 257:261 the invasive areas 244:261 the invasive areas of the tumor-stromal microenvironment in many human cancers 244:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 5 42 theme protein 1031:1037 arg1 interactions 1039:1050 other protein interactions 1025:1050 other protein interactions (vitronectin and integrins) 1025:1078 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 0 43 theme activator 53:61 arg1 receptor 63:70 the human urokinase plasminogen activator receptor 21:70 the human urokinase plasminogen activator receptor 21:70 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 5 44 theme receptor 993:1000 arg1 surface 1002:1008 the external receptor surface 980:1008 the external receptor surface accessible for other protein interactions (vitronectin and integrins) 980:1078 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 0 45 theme plasminogen 41:51 arg1 receptor 63:70 the human urokinase plasminogen activator receptor 21:70 the human urokinase plasminogen activator receptor 21:70 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 1 46 theme many 304:307 arg1 cancers 315:321 many human cancers 304:321 many human cancers 304:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 5 47 dep interactions 1039:1050 arg1 integrins 1069:1077 integrins 1069:1077 integrins 1069:1077 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 5 47 dep interactions 1039:1050 arg1 vitronectin 1053:1063 vitronectin 1053:1063 vitronectin 1053:1063 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 5 48 theme external 984:991 arg1 surface 1002:1008 the external receptor surface 980:1008 the external receptor surface accessible for other protein interactions (vitronectin and integrins) 980:1078 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 4 49 theme antagonist 789:798 arg1 peptide 800:806 the antagonist peptide 785:806 the antagonist peptide 785:806 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 15861141 1 50 theme human 309:313 arg1 cancers 315:321 many human cancers 304:321 many human cancers 304:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 4 51 theme uPA-uPAR 857:864 arg1 complex 866:872 the human uPA-uPAR complex 847:872 the human uPA-uPAR complex 847:872 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 15861141 3 52 theme helical 650:656 arg1 conformation 658:669 a helical conformation 648:669 a helical conformation 648:669 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 3 53 theme deep 600:603 arg1 cavity 614:619 both a deep internal cavity 593:619 both a deep internal cavity where the peptide binds in a helical conformation 593:669 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 1 54 theme crystal 118:124 arg1 structure 126:134 the crystal structure 114:134 the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers 114:321 We report the crystal structure of a soluble form of human urokinase-type plasminogen activator receptor (uPAR/CD87), which is expressed at the invasive areas of the tumor-stromal microenvironment in many human cancers. 15861141 3 55 theme three-finger 516:527 arg1 domains 529:535 three consecutive three-finger domains 498:535 three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, 498:670 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 3 56 theme internal 605:612 arg1 cavity 614:619 both a deep internal cavity 593:619 both a deep internal cavity where the peptide binds in a helical conformation 593:669 uPAR is composed of three consecutive three-finger domains organized in an almost circular manner, which generates both a deep internal cavity where the peptide binds in a helical conformation, and a large external surface. 15861141 4 57 theme binding 761:767 arg1 motif 769:773 a convergent binding motif 748:773 a convergent binding motif shared by the antagonist peptide and uPA 748:814 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 15861141 5 58 theme uPA 930:932 arg1 module 920:925 the receptor-binding module 899:925 the receptor-binding module of uPA 899:932 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 5 59 theme central 951:957 arg1 cavity 959:964 the uPAR central cavity 942:964 the uPAR central cavity 942:964 This model reveals that the receptor-binding module of uPA engages the uPAR central cavity, thus leaving the external receptor surface accessible for other protein interactions (vitronectin and integrins). 15861141 0 60 theme receptor 63:70 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human urokinase plasminogen activator receptor 0:70 Crystal structure of the human urokinase plasminogen activator receptor bound to an antagonist peptide. 15861141 6 61 theme unique 1089:1094 arg1 assembly 1107:1114 this unique structural assembly 1084:1114 this unique structural assembly 1084:1114 By this unique structural assembly, uPAR can orchestrate the fine interplay with the partners that are required to guide uPA-focalized proteolysis on the cell surface and control cell adhesion and migration. 15861141 4 62 theme motif 769:773 arg1 discovery 735:743 the discovery 731:743 the discovery of a convergent binding motif shared by the antagonist peptide and uPA 731:814 This knowledge combined with the discovery of a convergent binding motif shared by the antagonist peptide and uPA allowed us to build a model of the human uPA-uPAR complex. 30729188 5 0 contain contain 746:752 arg2 GM1 799:801 GM1 799:801 GM1 799:801 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 0 contain contain 746:752 arg2 lipids 773:778 negatively charged lipids 754:778 negatively charged lipids 754:778 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 0 contain contain 746:752 arg1 membranes 731:739 membranes 731:739 membranes that contain negatively charged lipids, including BMP and GM1 731:801 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 0 contain contain 746:752 arg2 BMP 791:793 BMP 791:793 BMP 791:793 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 1 1 theme ependymin-related 143:159 arg1 family 169:174 the ependymin-related protein family 139:174 the ependymin-related protein family 139:174 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 5 2 theme charged 765:771 arg1 GM1 799:801 GM1 799:801 GM1 799:801 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 2 theme charged 765:771 arg1 lipids 773:778 negatively charged lipids 754:778 negatively charged lipids 754:778 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 2 theme charged 765:771 arg1 BMP 791:793 BMP 791:793 BMP 791:793 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 1 3 theme protein 161:167 arg1 family 169:174 the ependymin-related protein family 139:174 the ependymin-related protein family 139:174 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 0 4 theme lipoprotein 94:104 arg1 transporters 106:117 bacterial lipoprotein transporters 84:117 bacterial lipoprotein transporters 84:117 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 5 5 theme lysosomal 848:856 arg1 protein 868:874 a lysosomal activator protein 846:874 a lysosomal activator protein 846:874 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 5 theme lysosomal 848:856 arg1 EPDR1 824:828 EPDR1 824:828 EPDR1 824:828 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 4 6 theme non-overlapping 635:649 arg1 grooves 663:669 two non-overlapping hydrophobic grooves 631:669 two non-overlapping hydrophobic grooves on the flat side of the hemisphere 631:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 1 7 theme most 258:261 arg1 vertebrates 263:273 most vertebrates 258:273 most vertebrates 258:273 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 0 8 theme bacterial 84:92 arg1 transporters 106:117 bacterial lipoprotein transporters 84:117 bacterial lipoprotein transporters 84:117 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 5 9 theme activator 858:866 arg1 protein 868:874 a lysosomal activator protein 846:874 a lysosomal activator protein 846:874 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 9 theme activator 858:866 arg1 EPDR1 824:828 EPDR1 824:828 EPDR1 824:828 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 4 10 from grooves 663:669 arg1 side 683:686 the flat side 674:686 the flat side of the hemisphere 674:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 1 11 theme vertebrates 263:273 arg1 lysosomes 230:238 lysosomes 230:238 lysosomes 230:238 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 1 11 theme vertebrates 263:273 arg1 secretomes 244:253 secretomes 244:253 secretomes 244:253 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 4 12 theme flat 678:681 arg1 side 683:686 the flat side 674:686 the flat side of the hemisphere 674:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 6 13 theme cellular 1051:1058 arg1 life 1060:1063 cellular life 1051:1063 cellular life 1051:1063 A phylogenetic analysis reveals that the fold is more widely distributed than previously suspected, with representatives identified in all branches of cellular life. 30729188 1 14 theme family 169:174 arg1 EPDR1 120:124 EPDR1 120:124 EPDR1 120:124 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 1 14 theme family 169:174 arg1 member 129:134 a member 127:134 a member of the ependymin-related protein family 127:174 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 0 15 theme transporters 106:117 arg1 superfamily 69:79 the superfamily 65:79 the superfamily of bacterial lipoprotein transporters 65:117 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 3 16 theme crystal 392:398 arg1 structures 400:409 crystal structures 392:409 crystal structures of human EPDR1 392:424 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 3 17 theme related 511:517 arg1 proteins 502:509 bacterial proteins 492:509 bacterial proteins related to the LolA lipoprotein transporter 492:553 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 6 18 theme life 1060:1063 arg1 branches 1039:1046 all branches 1035:1046 all branches of cellular life 1035:1063 A phylogenetic analysis reveals that the fold is more widely distributed than previously suspected, with representatives identified in all branches of cellular life. 30729188 0 19 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of human lysosomal EPDR1 0:42 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 4 20 theme hemisphere 695:704 arg1 side 683:686 the flat side 674:686 the flat side of the hemisphere 674:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 3 21 theme human 414:418 arg1 EPDR1 420:424 human EPDR1 414:424 human EPDR1 414:424 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 0 22 theme human 22:26 arg1 EPDR1 38:42 human lysosomal EPDR1 22:42 human lysosomal EPDR1 22:42 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 2 23 theme human 300:304 arg1 disease 306:312 human disease 300:312 human disease 300:312 Despite having roles in human disease and health, the molecular functions of EPDR1 remain unknown. 30729188 1 24 dep lysosomes 230:238 arg1 the 226:228 the 226:228 the 226:228 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 3 25 theme lipoprotein 531:541 arg1 transporter 543:553 the LolA lipoprotein transporter 522:553 the LolA lipoprotein transporter 522:553 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 1 26 theme uncharacterized 193:207 arg1 EPDR1 120:124 EPDR1 120:124 EPDR1 120:124 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 1 26 theme uncharacterized 193:207 arg1 protein 209:215 a relatively uncharacterized protein 180:215 a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates 180:273 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 0 27 theme EPDR1 38:42 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of human lysosomal EPDR1 0:42 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 4 28 theme hydrophobic 651:661 arg1 grooves 663:669 two non-overlapping hydrophobic grooves 631:669 two non-overlapping hydrophobic grooves on the flat side of the hemisphere 631:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 0 29 theme lysosomal 28:36 arg1 EPDR1 38:42 human lysosomal EPDR1 22:42 human lysosomal EPDR1 22:42 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 6 30 theme phylogenetic 902:913 arg1 analysis 915:922 A phylogenetic analysis 900:922 A phylogenetic analysis 900:922 A phylogenetic analysis reveals that the fold is more widely distributed than previously suspected, with representatives identified in all branches of cellular life. 30729188 3 31 theme EPDR1 420:424 arg1 structures 400:409 crystal structures 392:409 crystal structures of human EPDR1 392:424 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 2 32 theme molecular 330:338 arg1 functions 340:348 the molecular functions 326:348 the molecular functions of EPDR1 326:357 Despite having roles in human disease and health, the molecular functions of EPDR1 remain unknown. 30729188 4 33 with half-shell 615:624 arg1 grooves 663:669 two non-overlapping hydrophobic grooves 631:669 two non-overlapping hydrophobic grooves on the flat side of the hemisphere 631:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 4 34 with homodimer 570:578 arg1 shape 596:600 an overall shape 585:600 an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere 585:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 30729188 5 35 theme lipid 881:885 arg1 EPDR1 824:828 EPDR1 824:828 EPDR1 824:828 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 5 35 theme lipid 881:885 arg1 transporter 887:897 a lipid transporter 879:897 a lipid transporter 879:897 EPDR1 can interact with membranes that contain negatively charged lipids, including BMP and GM1, and we suggest that EPDR1 may function as a lysosomal activator protein or a lipid transporter. 30729188 3 36 theme LolA 526:529 arg1 transporter 543:553 the LolA lipoprotein transporter 522:553 the LolA lipoprotein transporter 522:553 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 1 37 located found 217:221 arg1 lysosomes 230:238 lysosomes 230:238 lysosomes 230:238 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 1 37 located found 217:221 arg2 EPDR1 120:124 EPDR1 120:124 EPDR1 120:124 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 1 37 located found 217:221 arg2 protein 209:215 a relatively uncharacterized protein 180:215 a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates 180:273 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 1 37 located found 217:221 arg1 secretomes 244:253 secretomes 244:253 secretomes 244:253 EPDR1, a member of the ependymin-related protein family, is a relatively uncharacterized protein found in the lysosomes and secretomes of most vertebrates. 30729188 0 38 with homology 51:58 arg1 superfamily 69:79 the superfamily 65:79 the superfamily of bacterial lipoprotein transporters 65:117 Crystal structures of human lysosomal EPDR1 reveal homology with the superfamily of bacterial lipoprotein transporters. 30729188 3 39 theme bacterial 492:500 arg1 proteins 502:509 bacterial proteins 492:509 bacterial proteins related to the LolA lipoprotein transporter 492:553 Here, we present crystal structures of human EPDR1 and reveal that the protein adopts a fold previously seen only in bacterial proteins related to the LolA lipoprotein transporter. 30729188 2 40 from roles 291:295 arg1 health 318:323 health 318:323 health 318:323 Despite having roles in human disease and health, the molecular functions of EPDR1 remain unknown. 30729188 2 40 from roles 291:295 arg1 disease 306:312 human disease 300:312 human disease 300:312 Despite having roles in human disease and health, the molecular functions of EPDR1 remain unknown. 30729188 2 41 theme EPDR1 353:357 arg1 functions 340:348 the molecular functions 326:348 the molecular functions of EPDR1 326:357 Despite having roles in human disease and health, the molecular functions of EPDR1 remain unknown. 30729188 4 42 theme overall 588:594 arg1 shape 596:600 an overall shape 585:600 an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere 585:704 EPDR1 forms a homodimer with an overall shape resembling a half-shell with two non-overlapping hydrophobic grooves on the flat side of the hemisphere. 28581485 5 0 from application 993:1003 arg1 enzymes 1065:1071 tracking pathologically relevant enzymes 1032:1071 tracking pathologically relevant enzymes 1032:1071 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 5 1 theme unanticipated 1152:1164 arg1 functionality 1193:1205 unanticipated structural and biochemical functionality 1152:1205 unanticipated structural and biochemical functionality 1152:1205 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 3 2 theme β-glucuronidase-specific 506:529 arg1 ABPs 554:557 ABPs 554:557 ABPs 554:557 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 2 theme β-glucuronidase-specific 506:529 arg1 probes 546:551 β-glucuronidase-specific activity-based probes 506:551 β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states 506:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 2 3 from activities 455:464 arg1 tissues 469:475 tissues 469:475 tissues 469:475 The medical importance of these enzymes necessitates reliable methods to assay their activities in tissues. 28581485 3 4 theme rapid 571:575 arg1 visualization 594:606 rapid and quantitative visualization 571:606 rapid and quantitative visualization of GUSB and HPSE 571:623 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 1 5 theme exo-acting 173:182 arg1 GUSB 201:204 GUSB 201:204 GUSB 201:204 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 5 theme exo-acting 173:182 arg1 β-glucuronidase 184:198 exo-acting β-glucuronidase 173:198 exo-acting β-glucuronidase (GUSB) 173:205 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 4 6 theme bacterial 945:953 arg1 homologs 955:962 their bacterial homologs 939:962 their bacterial homologs 939:962 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 4 7 theme structural 880:889 arg1 relationships 891:903 structural relationships 880:903 structural relationships between proHPSE, mature HPSE, and their bacterial homologs 880:962 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 1 8 theme endo-acting 275:285 arg1 HPSE 299:302 HPSE 299:302 HPSE 299:302 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 8 theme endo-acting 275:285 arg1 heparanase 287:296 endo-acting heparanase 275:296 endo-acting heparanase (HPSE) 275:303 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 8 theme endo-acting 275:285 arg1 enzymes 135:141 at least two distinct β-glucuronidase enzymes 97:141 at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII 97:268 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 9 dep enzymes 135:141 arg1 GUSB 201:204 GUSB 201:204 GUSB 201:204 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 9 dep enzymes 135:141 arg1 β-glucuronidase 184:198 exo-acting β-glucuronidase 173:198 exo-acting β-glucuronidase (GUSB) 173:205 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 2 10 theme enzymes 402:408 arg1 importance 382:391 The medical importance 370:391 The medical importance of these enzymes 370:408 The medical importance of these enzymes necessitates reliable methods to assay their activities in tissues. 28581485 3 11 theme disease 720:726 arg1 states 728:733 disease states 720:733 disease states 720:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 4 12 theme inactive 778:785 arg1 proHPSE 802:808 the supposedly inactive HPSE proenzyme proHPSE 763:808 the supposedly inactive HPSE proenzyme proHPSE 763:808 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 5 13 theme β-glucuronidase 1008:1022 arg1 ABPs 1024:1027 β-glucuronidase ABPs 1008:1027 β-glucuronidase ABPs 1008:1027 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 4 14 theme surprising 850:859 arg1 insights 861:868 surprising insights 850:868 surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs 850:962 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 0 15 theme Activity-based 0:13 arg1 probes 15:20 Activity-based probes 0:20 Activity-based probes for functional interrogation of retaining β-glucuronidases 0:79 Activity-based probes for functional interrogation of retaining β-glucuronidases. 28581485 5 16 theme structural 1166:1175 arg1 functionality 1193:1205 unanticipated structural and biochemical functionality 1152:1205 unanticipated structural and biochemical functionality 1152:1205 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 3 17 theme quantitative 581:592 arg1 visualization 594:606 rapid and quantitative visualization 571:606 rapid and quantitative visualization of GUSB and HPSE 571:623 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 0 18 theme functional 26:35 arg1 interrogation 37:49 functional interrogation 26:49 functional interrogation of retaining β-glucuronidases 26:79 Activity-based probes for functional interrogation of retaining β-glucuronidases. 28581485 5 19 theme ABPs 1024:1027 arg1 application 993:1003 the application 989:1003 the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes 989:1071 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 1 20 theme distinct 110:117 arg1 enzymes 135:141 at least two distinct β-glucuronidase enzymes 97:141 at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII 97:268 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 20 theme distinct 110:117 arg1 heparanase 287:296 endo-acting heparanase 275:296 endo-acting heparanase (HPSE) 275:303 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 5 21 theme tracking 1032:1039 arg1 enzymes 1065:1071 tracking pathologically relevant enzymes 1032:1071 tracking pathologically relevant enzymes 1032:1071 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 4 22 theme HPSE 787:790 arg1 proHPSE 802:808 the supposedly inactive HPSE proenzyme proHPSE 763:808 the supposedly inactive HPSE proenzyme proHPSE 763:808 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 1 23 theme β-glucuronidase 119:133 arg1 enzymes 135:141 at least two distinct β-glucuronidase enzymes 97:141 at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII 97:268 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 1 23 theme β-glucuronidase 119:133 arg1 heparanase 287:296 endo-acting heparanase 275:296 endo-acting heparanase (HPSE) 275:303 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 2 24 theme medical 374:380 arg1 importance 382:391 The medical importance 370:391 The medical importance of these enzymes 370:408 The medical importance of these enzymes necessitates reliable methods to assay their activities in tissues. 28581485 3 25 from activities 695:704 arg1 normal 709:714 normal 709:714 normal 709:714 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 25 from activities 695:704 arg1 states 728:733 disease states 720:733 disease states 720:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 26 theme powerful 660:667 arg1 tool 669:672 a powerful tool 658:672 a powerful tool for dissecting their activities in normal and disease states 658:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 27 theme GUSB 611:614 arg1 visualization 594:606 rapid and quantitative visualization 571:606 rapid and quantitative visualization of GUSB and HPSE 571:623 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 2 28 theme reliable 423:430 arg1 methods 432:438 reliable methods 423:438 reliable methods 423:438 The medical importance of these enzymes necessitates reliable methods to assay their activities in tissues. 28581485 0 29 theme β-glucuronidases 64:79 arg1 interrogation 37:49 functional interrogation 26:49 functional interrogation of retaining β-glucuronidases 26:79 Activity-based probes for functional interrogation of retaining β-glucuronidases. 28581485 5 30 theme biochemical 1181:1191 arg1 functionality 1193:1205 unanticipated structural and biochemical functionality 1152:1205 unanticipated structural and biochemical functionality 1152:1205 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 5 31 theme ABP-driven 1105:1114 arg1 approaches 1116:1125 ABP-driven approaches 1105:1125 ABP-driven approaches 1105:1125 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 5 32 theme relevant 1056:1063 arg1 enzymes 1065:1071 tracking pathologically relevant enzymes 1032:1071 tracking pathologically relevant enzymes 1032:1071 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 0 33 theme retaining 54:62 arg1 β-glucuronidases 64:79 retaining β-glucuronidases 54:79 retaining β-glucuronidases 54:79 Activity-based probes for functional interrogation of retaining β-glucuronidases. 28581485 5 34 theme functionality 1193:1205 arg1 discovery 1139:1147 discovery 1139:1147 discovery of unanticipated structural and biochemical functionality 1139:1205 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 4 35 theme mature 922:927 arg1 HPSE 929:932 mature HPSE 922:932 mature HPSE 922:932 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 3 36 theme biological 628:637 arg1 samples 639:645 biological samples 628:645 biological samples 628:645 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 4 37 theme proenzyme 792:800 arg1 proHPSE 802:808 the supposedly inactive HPSE proenzyme proHPSE 763:808 the supposedly inactive HPSE proenzyme proHPSE 763:808 Unexpectedly, we find that the supposedly inactive HPSE proenzyme proHPSE is also labeled by our ABPs, leading to surprising insights regarding structural relationships between proHPSE, mature HPSE, and their bacterial homologs. 28581485 5 38 theme case 1087:1090 arg1 study 1092:1096 a case study 1085:1096 a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality 1085:1205 Our results demonstrate the application of β-glucuronidase ABPs in tracking pathologically relevant enzymes and provide a case study of how ABP-driven approaches can lead to discovery of unanticipated structural and biochemical functionality. 28581485 3 39 theme HPSE 620:623 arg1 visualization 594:606 rapid and quantitative visualization 571:606 rapid and quantitative visualization of GUSB and HPSE 571:623 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 40 theme activity-based 531:544 arg1 ABPs 554:557 ABPs 554:557 ABPs 554:557 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 40 theme activity-based 531:544 arg1 probes 546:551 β-glucuronidase-specific activity-based probes 506:551 β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states 506:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 1 41 theme mucopolysaccharidosis 239:259 arg1 VII 266:268 mucopolysaccharidosis type VII 239:268 mucopolysaccharidosis type VII 239:268 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 28581485 3 42 theme probes 546:551 arg1 set 499:501 a set 497:501 a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states 497:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 42 theme probes 546:551 arg1 ABPs 554:557 ABPs 554:557 ABPs 554:557 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 3 42 theme probes 546:551 arg1 probes 546:551 β-glucuronidase-specific activity-based probes 506:551 β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states 506:733 Herein, we present a set of β-glucuronidase-specific activity-based probes (ABPs) that allow rapid and quantitative visualization of GUSB and HPSE in biological samples, providing a powerful tool for dissecting their activities in normal and disease states. 28581485 1 43 theme type 261:264 arg1 VII 266:268 mucopolysaccharidosis type VII 239:268 mucopolysaccharidosis type VII 239:268 Humans express at least two distinct β-glucuronidase enzymes that are involved in disease: exo-acting β-glucuronidase (GUSB), whose deficiency gives rise to mucopolysaccharidosis type VII, and endo-acting heparanase (HPSE), whose overexpression is implicated in inflammation and cancers. 16623661 0 0 theme rational 64:71 arg1 design 99:104 rational structure-based inhibitor design 64:104 rational structure-based inhibitor design 64:104 The structure of siglec-7 in complex with sialosides: leads for rational structure-based inhibitor design. 16623661 4 1 theme trimethylsilyl 946:959 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 1 theme trimethylsilyl 946:959 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 2 theme binding 1016:1022 arg1 site 1024:1027 the binding site 1012:1027 the binding site 1012:1027 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 6 3 theme alpha-methyl 1319:1330 arg1 scaffold 1376:1383 a basic scaffold 1368:1383 a basic scaffold 1368:1383 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 6 3 theme alpha-methyl 1319:1330 arg1 Neu5Ac 1332:1337 alpha-methyl Neu5Ac 1319:1337 alpha-methyl Neu5Ac (N-acetylneuraminic acid) 1319:1363 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 6 3 theme alpha-methyl 1319:1330 arg1 acid 1359:1362 N-acetylneuraminic acid 1340:1362 N-acetylneuraminic acid 1340:1362 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 4 4 theme detailed 988:995 arg1 description 997:1007 a detailed description 986:1007 a detailed description of the binding site, required for structure-guided inhibitor design 986:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 7 5 theme inhibition 1524:1533 arg1 data 1535:1538 inhibition data 1524:1538 inhibition data for the sialosides 1524:1557 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 5 theme inhibition 1524:1533 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 0 6 from structure 4:12 arg1 complex 29:35 complex 29:35 complex with sialosides 29:51 The structure of siglec-7 in complex with sialosides: leads for rational structure-based inhibitor design. 16623661 4 7 theme sialylated 846:855 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 7 theme sialylated 846:855 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 7 8 theme amino-oxalyl-amino 1484:1501 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 8 theme amino-oxalyl-amino 1484:1501 arg1 -9-deoxy-Neu5Ac 1503:1517 oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1452:1557 -9-deoxy-Neu5Ac 1503:1517 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 4 9 with complex 831:837 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 9 with complex 831:837 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 0 10 theme inhibitor 89:97 arg1 design 99:104 rational structure-based inhibitor design 64:104 rational structure-based inhibitor design 64:104 The structure of siglec-7 in complex with sialosides: leads for rational structure-based inhibitor design. 16623661 5 11 theme binding 1094:1100 arg1 assays 1102:1107 binding assays 1094:1107 binding assays 1094:1107 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 4 12 theme siglec-7 819:826 arg1 structure 806:814 the crystal structure 794:814 the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design 794:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 12 theme siglec-7 819:826 arg1 step 760:763 a first step 752:763 a first step towards this end 752:780 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 5 13 theme acid 1216:1219 arg1 binding 1221:1227 sialic acid binding 1209:1227 sialic acid binding 1209:1227 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 2 14 theme T-cells 528:534 arg1 subsets 513:519 subsets 513:519 subsets of CD8 T-cells 513:534 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 2 14 theme T-cells 528:534 arg1 monocytes 491:499 monocytes 491:499 monocytes 491:499 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 2 14 theme T-cells 528:534 arg1 cells 481:485 natural killer cells 466:485 natural killer cells 466:485 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 7 15 from co-crystal 1388:1397 arg1 complex 1414:1420 complex 1414:1420 complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1414:1557 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 16 theme inhibitor 1588:1596 arg1 design 1598:1603 future inhibitor design 1581:1603 future inhibitor design 1581:1603 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 17 theme siglec-7 1402:1409 arg1 co-crystal 1388:1397 A co-crystal 1386:1397 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1386:1557 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 1 18 theme haematopoietic 277:290 arg1 systems 312:318 the haematopoietic, immune and nervous systems 273:318 the haematopoietic, immune and nervous systems 273:318 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 2 19 theme CD8 524:526 arg1 T-cells 528:534 CD8 T-cells 524:534 CD8 T-cells 524:534 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 3 20 theme Siglec-specific 537:551 arg1 inhibitors 553:562 Siglec-specific inhibitors 537:562 Siglec-specific inhibitors 537:562 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 2 21 theme CD33-related 411:422 arg1 siglecs 424:430 immune inhibitory CD33-related siglecs 393:430 immune inhibitory CD33-related siglecs 393:430 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 1 22 gly sialylated 185:194 arg1 glycoconjugates 196:210 sialylated glycoconjugates 185:210 sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems 185:318 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 6 23 theme basic 1370:1374 arg1 Neu5Ac 1332:1337 alpha-methyl Neu5Ac 1319:1337 alpha-methyl Neu5Ac (N-acetylneuraminic acid) 1319:1363 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 6 23 theme basic 1370:1374 arg1 scaffold 1376:1383 a basic scaffold 1368:1383 a basic scaffold 1368:1383 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 1 24 theme transmembrane 157:169 arg1 receptors 171:179 transmembrane receptors 157:179 transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems 157:318 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 1 24 theme transmembrane 157:169 arg1 Siglecs 107:113 Siglecs 107:113 Siglecs (sialic acid binding Ig-like lectins) 107:151 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 2 25 theme killer 474:479 arg1 cells 481:485 natural killer cells 466:485 natural killer cells 466:485 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 3 26 theme potential 718:726 arg1 siglecs 707:713 siglecs 707:713 siglecs 707:713 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 3 26 theme potential 718:726 arg1 targets 740:746 potential therapeutic targets 718:746 potential therapeutic targets 718:746 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 7 27 dep clear 1565:1569 arg1 leads 1571:1575 leads 1571:1575 leads for future inhibitor design 1571:1603 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 2 28 theme immune 393:398 arg1 siglecs 424:430 immune inhibitory CD33-related siglecs 393:430 immune inhibitory CD33-related siglecs 393:430 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 2 29 theme natural 466:472 arg1 killer 474:479 natural killer 466:479 natural killer cells 466:485 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 7 30 theme oxamido-Neu5Ac 1452:1465 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 30 theme oxamido-Neu5Ac 1452:1465 arg1 -9-deoxy-Neu5Ac 1503:1517 oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1452:1557 -9-deoxy-Neu5Ac 1503:1517 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 4 31 theme structure-guided 1043:1058 arg1 design 1070:1075 structure-guided inhibitor design 1043:1075 structure-guided inhibitor design 1043:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 0 32 theme siglec-7 17:24 arg1 structure 4:12 The structure 0:12 The structure of siglec-7 in complex with sialosides: 0:52 The structure of siglec-7 in complex with sialosides: leads for rational structure-based inhibitor design. 16623661 1 33 theme immune 293:298 arg1 systems 312:318 the haematopoietic, immune and nervous systems 273:318 the haematopoietic, immune and nervous systems 273:318 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 4 34 theme 2,3 903:905 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 34 theme 2,3 903:905 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 34 theme 2,3 903:905 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 35 theme first 754:758 arg1 structure 806:814 the crystal structure 794:814 the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design 794:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 35 theme first 754:758 arg1 step 760:763 a first step 752:763 a first step towards this end 752:780 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 1 36 theme sialylated 185:194 arg1 glycoconjugates 196:210 sialylated glycoconjugates 185:210 sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems 185:318 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 5 37 theme key 1136:1138 arg1 role 1151:1154 a key structural role 1134:1154 a key structural role for Lys131, a residue that changes conformation upon sialic acid binding 1134:1227 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 6 38 theme members 1285:1291 arg1 sites 1262:1266 the binding sites 1250:1266 the binding sites of siglec family members 1250:1291 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 6 38 theme members 1285:1291 arg1 members 1285:1291 siglec family members 1271:1291 siglec family members 1271:1291 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 5 39 theme sialic 1209:1214 arg1 binding 1221:1227 sialic acid binding 1209:1227 sialic acid binding 1209:1227 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 5 40 theme structural 1140:1149 arg1 role 1151:1154 a key structural role 1134:1154 a key structural role for Lys131, a residue that changes conformation upon sialic acid binding 1134:1227 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 1 41 theme nervous 304:310 arg1 systems 312:318 the haematopoietic, immune and nervous systems 273:318 the haematopoietic, immune and nervous systems 273:318 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 6 42 theme family 1278:1283 arg1 members 1285:1291 siglec family members 1271:1291 siglec family members 1271:1291 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 5 43 used used 1114:1117 arg2 assays 1102:1107 binding assays 1094:1107 binding assays 1094:1107 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 5 43 used used 1114:1117 arg2 Mutagenesis 1078:1088 Mutagenesis 1078:1088 Mutagenesis 1078:1088 Mutagenesis and binding assays were used to demonstrate a key structural role for Lys131, a residue that changes conformation upon sialic acid binding. 16623661 4 44 theme inhibitor 1060:1068 arg1 design 1070:1075 structure-guided inhibitor design 1043:1075 structure-guided inhibitor design 1043:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 45 gly sialylated 846:855 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 45 gly sialylated 846:855 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 6 46 theme siglec 1271:1276 arg1 members 1285:1291 siglec family members 1271:1291 siglec family members 1271:1291 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 4 47 theme crystal 798:804 arg1 structure 806:814 the crystal structure 794:814 the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design 794:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 47 theme crystal 798:804 arg1 step 760:763 a first step 752:763 a first step towards this end 752:780 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 2 48 theme siglecs 424:430 arg1 family 383:388 the recently described family 360:388 the recently described family of immune inhibitory CD33-related siglecs 360:430 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 3 49 dep potential 718:726 arg1 therapeutic 728:738 therapeutic 728:738 therapeutic 728:738 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 4 50 theme analogue 881:888 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 50 theme analogue 881:888 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 50 theme analogue 881:888 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 6 51 theme N-acetylneuraminic 1340:1357 arg1 Neu5Ac 1332:1337 alpha-methyl Neu5Ac 1319:1337 alpha-methyl Neu5Ac (N-acetylneuraminic acid) 1319:1363 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 6 51 theme N-acetylneuraminic 1340:1357 arg1 acid 1359:1362 N-acetylneuraminic acid 1340:1362 N-acetylneuraminic acid 1340:1362 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 3 52 theme masked 597:602 arg1 detection 584:592 the detection 580:592 the detection of masked 580:602 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 6 53 theme binding 1254:1260 arg1 sites 1262:1266 the binding sites 1250:1266 the binding sites of siglec family members 1250:1291 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 6 53 theme binding 1254:1260 arg1 members 1285:1291 siglec family members 1271:1291 siglec family members 1271:1291 Differences between the binding sites of siglec family members were then exploited using alpha-methyl Neu5Ac (N-acetylneuraminic acid) as a basic scaffold. 16623661 2 54 theme described 373:381 arg1 family 383:388 the recently described family 360:388 the recently described family of immune inhibitory CD33-related siglecs 360:430 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 4 55 theme ganglioside 869:879 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 55 theme ganglioside 869:879 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 55 theme ganglioside 869:879 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 7 56 theme [methyl 1467:1473 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 56 theme [methyl 1467:1473 arg1 -9-deoxy-Neu5Ac 1503:1517 oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1452:1557 -9-deoxy-Neu5Ac 1503:1517 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 57 with complex 1414:1420 arg1 data 1535:1538 inhibition data 1524:1538 inhibition data for the sialosides 1524:1557 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 57 with complex 1414:1420 arg1 -9-deoxy-Neu5Ac 1503:1517 oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1452:1557 -9-deoxy-Neu5Ac 1503:1517 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 57 with complex 1414:1420 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 4 58 from structure 806:814 arg1 complex 831:837 complex 831:837 complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design 831:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 59 theme disialyl 919:926 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 59 theme disialyl 919:926 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 3 60 theme siglecs 626:632 arg1 forms 617:621 forms 617:621 forms of siglecs 617:632 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 4 61 theme [alpha 896:901 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 61 theme [alpha 896:901 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 61 theme [alpha 896:901 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 7 62 theme alpha-9- 1475:1482 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 62 theme alpha-9- 1475:1482 arg1 -9-deoxy-Neu5Ac 1503:1517 oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1452:1557 -9-deoxy-Neu5Ac 1503:1517 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 1 63 theme sialic 116:121 arg1 Siglecs 107:113 Siglecs 107:113 Siglecs (sialic acid binding Ig-like lectins) 107:151 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 1 63 theme sialic 116:121 arg1 lectins 144:150 sialic acid binding Ig-like lectins 116:150 sialic acid binding Ig-like lectins 116:150 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 4 64 theme site 1024:1027 arg1 description 997:1007 a detailed description 986:1007 a detailed description of the binding site, required for structure-guided inhibitor design 986:1075 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 1 65 theme cellular 226:233 arg1 interactions 235:246 cellular interactions 226:246 cellular interactions 226:246 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 4 66 theme DSLc4 890:894 arg1 ligand 857:862 a sialylated ligand 844:862 a sialylated ligand 844:862 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 66 theme DSLc4 890:894 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 66 theme DSLc4 890:894 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 3 67 theme signalling 659:668 arg1 pathways 670:677 signalling pathways 659:677 signalling pathways 659:677 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 7 68 theme future 1581:1586 arg1 design 1598:1603 future inhibitor design 1581:1603 future inhibitor design 1581:1603 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 1 69 theme acid 123:126 arg1 Siglecs 107:113 Siglecs 107:113 Siglecs (sialic acid binding Ig-like lectins) 107:151 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 1 69 theme acid 123:126 arg1 lectins 144:150 sialic acid binding Ig-like lectins 116:150 sialic acid binding Ig-like lectins 116:150 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 2 70 theme structural 335:344 arg1 prototype 346:354 a structural prototype 333:354 a structural prototype for the recently described family of immune inhibitory CD33-related siglecs 333:430 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 2 70 theme structural 335:344 arg1 Siglec-7 321:328 Siglec-7 321:328 Siglec-7 321:328 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 3 71 theme pathways 670:677 arg1 dissection 645:654 dissection 645:654 dissection of signalling pathways 645:677 Siglec-specific inhibitors are desired for the detection of masked and unmasked forms of siglecs, to aid in dissection of signalling pathways and as tools to investigate siglecs as potential therapeutic targets. 16623661 1 72 theme binding 128:134 arg1 Siglecs 107:113 Siglecs 107:113 Siglecs (sialic acid binding Ig-like lectins) 107:151 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 1 72 theme binding 128:134 arg1 lectins 144:150 sialic acid binding Ig-like lectins 116:150 sialic acid binding Ig-like lectins 116:150 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 2 73 theme inhibitory 400:409 arg1 siglecs 424:430 immune inhibitory CD33-related siglecs 393:430 immune inhibitory CD33-related siglecs 393:430 Siglec-7 is a structural prototype for the recently described family of immune inhibitory CD33-related siglecs and is predominantly expressed on natural killer cells and monocytes, as well as subsets of CD8 T-cells. 16623661 4 74 theme lactotetraosyl 928:941 arg1 ethyl 961:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl 914:965 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 4 74 theme lactotetraosyl 928:941 arg1 /alpha 907:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha 865:912 the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl] 865:966 As a first step towards this end, we present the crystal structure of siglec-7 in complex with a sialylated ligand, the ganglioside analogue DSLc4 [alpha(2,3)/alpha(2,6) disialyl lactotetraosyl 2-(trimethylsilyl)ethyl], which allows for a detailed description of the binding site, required for structure-guided inhibitor design. 16623661 7 75 theme sialoside 1431:1439 arg1 data 1535:1538 inhibition data 1524:1538 inhibition data for the sialosides 1524:1557 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 75 theme sialoside 1431:1439 arg1 -9-deoxy-Neu5Ac 1503:1517 oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides 1452:1557 -9-deoxy-Neu5Ac 1503:1517 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 7 75 theme sialoside 1431:1439 arg1 inhibitor 1441:1449 the sialoside inhibitor 1427:1449 the sialoside inhibitor 1427:1449 A co-crystal of siglec-7 in complex with the sialoside inhibitor, oxamido-Neu5Ac [methyl alpha-9-(amino-oxalyl-amino)-9-deoxy-Neu5Ac] and inhibition data for the sialosides gives clear leads for future inhibitor design. 16623661 1 76 theme signalling 252:261 arg1 events 263:268 signalling events 252:268 signalling events 252:268 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 0 77 with complex 29:35 arg1 sialosides 42:51 sialosides 42:51 sialosides 42:51 The structure of siglec-7 in complex with sialosides: leads for rational structure-based inhibitor design. 16623661 1 78 theme Ig-like 136:142 arg1 Siglecs 107:113 Siglecs 107:113 Siglecs (sialic acid binding Ig-like lectins) 107:151 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 1 78 theme Ig-like 136:142 arg1 lectins 144:150 sialic acid binding Ig-like lectins 116:150 sialic acid binding Ig-like lectins 116:150 Siglecs (sialic acid binding Ig-like lectins) are transmembrane receptors for sialylated glycoconjugates that modulate cellular interactions and signalling events in the haematopoietic, immune and nervous systems. 16623661 0 79 theme structure-based 73:87 arg1 design 99:104 rational structure-based inhibitor design 64:104 rational structure-based inhibitor design 64:104 The structure of siglec-7 in complex with sialosides: leads for rational structure-based inhibitor design. 16895906 7 0 theme preferred 1218:1226 arg1 conformations 1237:1249 preferred solution conformations 1218:1249 preferred solution conformations of GT1b 1218:1257 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 7 1 theme GT1b 1254:1257 arg1 conformations 1237:1249 preferred solution conformations 1218:1249 preferred solution conformations of GT1b 1218:1257 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 7 2 theme solution 1228:1235 arg1 conformations 1237:1249 preferred solution conformations 1218:1249 preferred solution conformations of GT1b 1218:1257 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 5 3 theme crystal 601:607 arg1 structure 609:617 The crystal structure 597:617 The crystal structure of the complex 597:632 The crystal structure of the complex offers a first glimpse into how this important family of lectins binds the structurally diverse gangliosides. 16895906 1 4 theme mammalian 137:145 arg1 receptors 167:175 mammalian sialic acid binding receptors 137:175 mammalian sialic acid binding receptors expressed predominantly in the immune system 137:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 4 5 theme ganglioside 579:589 arg1 GT1b 591:594 the alpha(2,8)-disialylated ganglioside GT1b 551:594 the alpha(2,8)-disialylated ganglioside GT1b 551:594 We have co-crystallized Siglec-7 with a synthetic oligosaccharide corresponding to the alpha(2,8)-disialylated ganglioside GT1b. 16895906 4 6 theme synthetic 508:516 arg1 oligosaccharide 518:532 a synthetic oligosaccharide 506:532 a synthetic oligosaccharide corresponding to the alpha(2,8)-disialylated ganglioside GT1b 506:594 We have co-crystallized Siglec-7 with a synthetic oligosaccharide corresponding to the alpha(2,8)-disialylated ganglioside GT1b. 16895906 7 7 theme hydrophobic 1104:1114 arg1 contacts 1116:1123 extensive hydrophobic contacts 1094:1123 extensive hydrophobic contacts together with key polar interactions 1094:1160 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 2 8 theme sialylated 286:295 arg1 glycans 297:303 sialylated glycans 286:303 sialylated glycans 286:303 The CD33-related siglecs show complex recognition patterns for sialylated glycans. 16895906 7 9 with combination 1007:1017 arg1 studies 1036:1042 mutagenesis studies 1024:1042 mutagenesis studies 1024:1042 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 2 10 gly sialylated 286:295 arg1 glycans 297:303 sialylated glycans 286:303 sialylated glycans 286:303 The CD33-related siglecs show complex recognition patterns for sialylated glycans. 16895906 6 11 theme ganglioside 971:981 arg1 core 959:962 the underlying neutral glycan core 929:962 the underlying neutral glycan core of the ganglioside 929:981 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 5 12 theme complex 626:632 arg1 structure 609:617 The crystal structure 597:617 The crystal structure of the complex 597:632 The crystal structure of the complex offers a first glimpse into how this important family of lectins binds the structurally diverse gangliosides. 16895906 1 13 theme sialic 147:152 arg1 receptors 167:175 mammalian sialic acid binding receptors 137:175 mammalian sialic acid binding receptors expressed predominantly in the immune system 137:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 0 14 theme -disialylganglioside 83:102 arg1 GT1b 104:107 the alpha(2,8)-disialylganglioside GT1b 69:107 the alpha(2,8)-disialylganglioside GT1b 69:107 Siglec-7 undergoes a major conformational change when complexed with the alpha(2,8)-disialylganglioside GT1b. 16895906 6 15 theme conformational 878:891 arg1 shift 893:897 a dramatic conformational shift 867:897 a dramatic conformational shift 867:897 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 7 16 theme extensive 1094:1102 arg1 contacts 1116:1123 extensive hydrophobic contacts 1094:1123 extensive hydrophobic contacts together with key polar interactions 1094:1160 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 2 17 theme recognition 261:271 arg1 patterns 273:280 complex recognition patterns 253:280 complex recognition patterns for sialylated glycans 253:303 The CD33-related siglecs show complex recognition patterns for sialylated glycans. 16895906 1 18 theme acid 154:157 arg1 receptors 167:175 mammalian sialic acid binding receptors 137:175 mammalian sialic acid binding receptors expressed predominantly in the immune system 137:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 7 19 theme binding 1175:1181 arg1 complementary 1201:1213 complementary 1201:1213 complementary 1201:1213 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 7 19 theme binding 1175:1181 arg1 structure 1188:1196 the binding site structure 1171:1196 the binding site structure 1171:1196 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 6 20 theme dramatic 869:876 arg1 shift 893:897 a dramatic conformational shift 867:897 a dramatic conformational shift 867:897 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 1 21 theme binding 159:165 arg1 receptors 167:175 mammalian sialic acid binding receptors 137:175 mammalian sialic acid binding receptors expressed predominantly in the immune system 137:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 7 22 theme site 1183:1186 arg1 complementary 1201:1213 complementary 1201:1213 complementary 1201:1213 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 7 22 theme site 1183:1186 arg1 structure 1188:1196 the binding site structure 1171:1196 the binding site structure 1171:1196 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 5 23 theme first 643:647 arg1 glimpse 649:655 a first glimpse 641:655 a first glimpse into how this important family of lectins binds the structurally diverse gangliosides 641:741 The crystal structure of the complex offers a first glimpse into how this important family of lectins binds the structurally diverse gangliosides. 16895906 3 24 theme key 422:424 arg1 interactions 426:437 the key interactions 418:437 the key interactions that drive this selectivity 418:465 Siglec-7 shows a preference for alpha(2,8)-disialylated ligands and provides a structural template for studying the key interactions that drive this selectivity. 16895906 0 25 theme major 21:25 arg1 change 42:47 a major conformational change 19:47 a major conformational change when complexed with the alpha(2,8)-disialylganglioside GT1b 19:107 Siglec-7 undergoes a major conformational change when complexed with the alpha(2,8)-disialylganglioside GT1b. 16895906 3 26 gly -disialylated 348:360 arg1 ligands 362:368 alpha(2,8)-disialylated ligands 338:368 alpha(2,8)-disialylated ligands 338:368 Siglec-7 shows a preference for alpha(2,8)-disialylated ligands and provides a structural template for studying the key interactions that drive this selectivity. 16895906 6 27 theme previous 809:816 arg1 studies 818:824 previous studies 809:824 previous studies 809:824 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 7 28 from data 999:1002 arg1 combination 1007:1017 combination 1007:1017 combination with mutagenesis studies 1007:1042 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 4 29 theme -disialylated 565:577 arg1 GT1b 591:594 the alpha(2,8)-disialylated ganglioside GT1b 551:594 the alpha(2,8)-disialylated ganglioside GT1b 551:594 We have co-crystallized Siglec-7 with a synthetic oligosaccharide corresponding to the alpha(2,8)-disialylated ganglioside GT1b. 16895906 7 30 theme ganglioside 1069:1079 arg1 binding 1054:1060 binding 1054:1060 binding of the ganglioside 1054:1079 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 6 31 theme glycan 952:957 arg1 core 959:962 the underlying neutral glycan core 929:962 the underlying neutral glycan core of the ganglioside 929:981 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 1 32 theme receptors 167:175 arg1 siglecs 114:120 The siglecs 110:120 The siglecs 110:120 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 1 32 theme receptors 167:175 arg1 group 128:132 a group 126:132 a group of mammalian sialic acid binding receptors expressed predominantly in the immune system 126:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 1 32 theme receptors 167:175 arg1 receptors 167:175 mammalian sialic acid binding receptors 137:175 mammalian sialic acid binding receptors expressed predominantly in the immune system 137:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 0 33 theme conformational 27:40 arg1 change 42:47 a major conformational change 19:47 a major conformational change when complexed with the alpha(2,8)-disialylganglioside GT1b 19:107 Siglec-7 undergoes a major conformational change when complexed with the alpha(2,8)-disialylganglioside GT1b. 16895906 6 34 theme neutral 944:950 arg1 core 959:962 the underlying neutral glycan core 929:962 the underlying neutral glycan core of the ganglioside 929:981 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 5 35 theme diverse 722:728 arg1 gangliosides 730:741 the structurally diverse gangliosides 705:741 the structurally diverse gangliosides 705:741 The crystal structure of the complex offers a first glimpse into how this important family of lectins binds the structurally diverse gangliosides. 16895906 6 36 theme C-C 775:777 arg1 loop 780:783 the C-C' loop 771:783 the C-C' loop 771:783 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 6 36 theme C-C 775:777 arg1 region 788:793 a region 786:793 a region implicated in previous studies as driving siglec specificity 786:854 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 3 37 theme structural 385:394 arg1 template 396:403 a structural template 383:403 a structural template for studying the key interactions that drive this selectivity 383:465 Siglec-7 shows a preference for alpha(2,8)-disialylated ligands and provides a structural template for studying the key interactions that drive this selectivity. 16895906 6 38 theme underlying 933:942 arg1 core 959:962 the underlying neutral glycan core 929:962 the underlying neutral glycan core of the ganglioside 929:981 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 5 39 theme important 671:679 arg1 family 681:686 this important family 666:686 this important family of lectins 666:697 The crystal structure of the complex offers a first glimpse into how this important family of lectins binds the structurally diverse gangliosides. 16895906 7 40 theme polar 1143:1147 arg1 interactions 1149:1160 key polar interactions 1139:1160 key polar interactions 1139:1160 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 6 41 theme siglec 837:842 arg1 specificity 844:854 siglec specificity 837:854 siglec specificity 837:854 The structure reveals that the C-C' loop, a region implicated in previous studies as driving siglec specificity, undergoes a dramatic conformational shift, allowing it to interact with the underlying neutral glycan core of the ganglioside. 16895906 7 42 theme mutagenesis 1024:1034 arg1 studies 1036:1042 mutagenesis studies 1024:1042 mutagenesis studies 1024:1042 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 2 43 theme complex 253:259 arg1 patterns 273:280 complex recognition patterns 253:280 complex recognition patterns for sialylated glycans 253:303 The CD33-related siglecs show complex recognition patterns for sialylated glycans. 16895906 4 44 gly -disialylated 565:577 arg1 GT1b 591:594 the alpha(2,8)-disialylated ganglioside GT1b 551:594 the alpha(2,8)-disialylated ganglioside GT1b 551:594 We have co-crystallized Siglec-7 with a synthetic oligosaccharide corresponding to the alpha(2,8)-disialylated ganglioside GT1b. 16895906 1 45 theme immune 208:213 arg1 system 215:220 the immune system 204:220 the immune system 204:220 The siglecs are a group of mammalian sialic acid binding receptors expressed predominantly in the immune system. 16895906 5 46 theme lectins 691:697 arg1 family 681:686 this important family 666:686 this important family of lectins 666:697 The crystal structure of the complex offers a first glimpse into how this important family of lectins binds the structurally diverse gangliosides. 16895906 7 47 theme structural 988:997 arg1 data 999:1002 The structural data 984:1002 The structural data in combination with mutagenesis studies 984:1042 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 7 48 theme key 1139:1141 arg1 interactions 1149:1160 key polar interactions 1139:1160 key polar interactions 1139:1160 The structural data in combination with mutagenesis studies show that binding of the ganglioside is driven by extensive hydrophobic contacts together with key polar interactions and that the binding site structure is complementary to preferred solution conformations of GT1b. 16895906 2 49 theme CD33-related 227:238 arg1 siglecs 240:246 The CD33-related siglecs 223:246 The CD33-related siglecs 223:246 The CD33-related siglecs show complex recognition patterns for sialylated glycans. 16895906 3 50 theme -disialylated 348:360 arg1 ligands 362:368 alpha(2,8)-disialylated ligands 338:368 alpha(2,8)-disialylated ligands 338:368 Siglec-7 shows a preference for alpha(2,8)-disialylated ligands and provides a structural template for studying the key interactions that drive this selectivity. 1569188 3 0 theme deglycosylated 507:520 arg1 centrifugation 479:492 cesium chloride gradient centrifugation 454:492 cesium chloride gradient centrifugation 454:492 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 13 1 from cartilage 1599:1607 arg1 release 1554:1560 the release 1550:1560 the release of aggrecan fragments from articular cartilage into the synovial fluid 1550:1631 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 13 1 from cartilage 1599:1607 arg1 fragments 1574:1582 aggrecan fragments 1565:1582 aggrecan fragments from articular cartilage 1565:1607 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 6 2 theme 374 904:906 arg1 bond 908:911 the Glu 373-Ala 374 bond 888:911 the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains 888:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 4 3 theme aggrecan 605:612 arg1 populations 627:637 two major aggrecan core protein populations 595:637 two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD 595:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 6 4 theme NH2 815:817 arg1 terminus 819:826 This NH2 terminus 810:826 This NH2 terminus 810:826 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 6 5 theme Glu 892:894 arg1 bond 908:911 the Glu 373-Ala 374 bond 888:911 the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains 888:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 12 6 from present 1256:1262 arg1 fluid 1302:1306 both osteoarthritic human synovial fluid 1267:1306 both osteoarthritic human synovial fluid 1267:1306 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 6 from present 1256:1262 arg1 fluid 1338:1342 normal bovine synovial fluid 1315:1342 normal bovine synovial fluid 1315:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 2 7 theme knee 253:256 arg1 injury 258:263 recent knee injury 246:263 recent knee injury 246:263 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 1 8 theme 374 160:162 arg1 bond 164:167 the Glu 373-Ala 374 bond 144:167 the Glu 373-Ala 374 bond of the interglobular domain 144:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 7 9 theme explant 1094:1100 arg1 cultures 1102:1109 bovine cartilage explant cultures 1077:1109 bovine cartilage explant cultures 1077:1109 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 2 10 theme early 288:292 arg1 osteoarthritis 308:321 early or late stage osteoarthritis 288:321 early or late stage osteoarthritis 288:321 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 16 11 theme proteinase 1780:1789 arg1 action 1751:1756 the action 1747:1756 the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain 1747:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 13 12 dep seen 1633:1636 arg1 L. 1688:1689 L. 1688:1689 L. 1688:1689 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 13 12 dep seen 1633:1636 arg1 S. 1691:1692 S. 1691:1692 S. 1691:1692 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 6 13 theme G2 964:965 arg1 domains 967:973 the G1 and G2 domains 953:973 the G1 and G2 domains 953:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 4 14 theme molecular 653:661 arg1 masses 663:668 apparent molecular masses 644:668 apparent molecular masses of approximately 90 kD and 150 kD 644:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 7 15 theme bovine 1077:1082 arg1 cultures 1102:1109 bovine cartilage explant cultures 1077:1109 bovine cartilage explant cultures 1077:1109 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 1 16 theme interglobular 176:188 arg1 domain 190:195 the interglobular domain 172:195 the interglobular domain 172:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 6 17 theme G1 957:958 arg1 domains 967:973 the G1 and G2 domains 953:973 the G1 and G2 domains 953:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 12 18 theme normal 1315:1320 arg1 fluid 1338:1342 normal bovine synovial fluid 1315:1342 normal bovine synovial fluid 1315:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 13 19 theme human 1655:1659 arg1 osteoarthritis 1661:1674 human osteoarthritis 1655:1674 human osteoarthritis 1655:1674 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 5 20 theme NH2-terminal 722:733 arg1 analysis 735:742 NH2-terminal analysis 722:742 NH2-terminal analysis of both populations 722:762 For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. 1569188 4 21 with populations 627:637 arg1 masses 663:668 apparent molecular masses 644:668 apparent molecular masses of approximately 90 kD and 150 kD 644:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 12 22 theme s 1529:1529 arg1 domain 1438:1443 the G2 domain 1431:1443 the G2 domain 1431:1443 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 22 theme s 1529:1529 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 22 theme s 1529:1529 arg1 lengths 1487:1493 variable lengths 1478:1493 variable lengths of the chondroitin sulfate domain(s) 1478:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 22 theme s 1529:1529 arg1 domain 1423:1428 the interglobular domain 1405:1428 the interglobular domain 1405:1428 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 22 theme s 1529:1529 arg1 domain 1466:1471 the keratan sulfate domain 1446:1471 the keratan sulfate domain 1446:1471 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 23 theme domain 1522:1527 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 24 theme synovial 1329:1336 arg1 fluid 1338:1342 normal bovine synovial fluid 1315:1342 normal bovine synovial fluid 1315:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 25 theme domain 1423:1428 arg1 stretch 1394:1400 a short NH2-terminal stretch 1373:1400 a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s) 1373:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 7 26 theme stimulated 1043:1052 arg1 catabolism 1063:1072 interleukin-1 stimulated aggrecan catabolism 1029:1072 interleukin-1 stimulated aggrecan catabolism 1029:1072 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 2 27 theme Synovial 198:205 arg1 fluid 207:211 Synovial fluid 198:211 Synovial fluid 198:211 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 6 28 theme interglobular 924:936 arg1 domain 938:943 the interglobular domain 920:943 the interglobular domain between the G1 and G2 domains 920:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 12 29 theme synovial 1293:1300 arg1 fluid 1302:1306 both osteoarthritic human synovial fluid 1267:1306 both osteoarthritic human synovial fluid 1267:1306 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 16 30 theme 373-Ala 1813:1819 arg1 bond 1825:1828 the Glu 373-Ala 374 bond 1805:1828 the Glu 373-Ala 374 bond of the interglobular domain 1805:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 1 31 theme proteinase 119:128 arg1 osteoarthritis 93:106 osteoarthritis 93:106 osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain 93:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 3 32 attach present 375:381 arg2 fragments 365:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 32 attach present 375:381 arg1 fluid 430:434 normal bovine synovial fluid 407:434 normal bovine synovial fluid 407:434 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 32 attach present 375:381 arg1 fluids 392:397 these fluids 386:397 these fluids 386:397 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 4 33 theme kD 701:702 arg1 masses 663:668 apparent molecular masses 644:668 apparent molecular masses of approximately 90 kD and 150 kD 644:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 12 34 theme chondroitin 1502:1512 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 35 attach present 1256:1262 arg1 fluid 1302:1306 both osteoarthritic human synovial fluid 1267:1306 both osteoarthritic human synovial fluid 1267:1306 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 35 attach present 1256:1262 arg1 fluid 1338:1342 normal bovine synovial fluid 1315:1342 normal bovine synovial fluid 1315:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 35 attach present 1256:1262 arg2 fragments 1246:1254 the major aggrecan fragments 1227:1254 the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid 1227:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 35 attach present 1256:1262 arg2 large 1348:1352 large 1348:1352 large 1348:1352 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 36 theme NH2-terminal 1381:1392 arg1 stretch 1394:1400 a short NH2-terminal stretch 1373:1400 a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s) 1373:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 5 37 theme major 778:782 arg1 ARGSV 803:807 a single major sequence beginning ARGSV 769:807 a single major sequence beginning ARGSV 769:807 For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. 1569188 1 38 theme Glu 148:150 arg1 bond 164:167 the Glu 373-Ala 374 bond 144:167 the Glu 373-Ala 374 bond of the interglobular domain 144:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 2 39 with patients 274:281 arg1 osteoarthritis 308:321 early or late stage osteoarthritis 288:321 early or late stage osteoarthritis 288:321 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 3 40 theme aggrecan 356:363 arg1 fragments 365:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 41 from present 375:381 arg1 fluid 430:434 normal bovine synovial fluid 407:434 normal bovine synovial fluid 407:434 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 41 from present 375:381 arg1 fluids 392:397 these fluids 386:397 these fluids 386:397 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 12 42 theme variable 1478:1485 arg1 lengths 1487:1493 variable lengths 1478:1493 variable lengths of the chondroitin sulfate domain(s) 1478:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 43 theme Chondroitin 324:334 arg1 fragments 365:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 44 gly deglycosylated 507:520 arg1 centrifugation 479:492 cesium chloride gradient centrifugation 454:492 cesium chloride gradient centrifugation 454:492 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 5 45 theme populations 752:762 arg1 analysis 735:742 NH2-terminal analysis 722:742 NH2-terminal analysis of both populations 722:762 For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. 1569188 12 46 theme present 1256:1262 arg1 large 1348:1352 large 1348:1352 large 1348:1352 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 46 theme present 1256:1262 arg1 fragments 1246:1254 the major aggrecan fragments 1227:1254 the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid 1227:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 13 47 theme synovial 1618:1625 arg1 fluid 1627:1631 the synovial fluid 1614:1631 the synovial fluid 1614:1631 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 12 48 theme short 1375:1379 arg1 stretch 1394:1400 a short NH2-terminal stretch 1373:1400 a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s) 1373:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 49 theme bovine 414:419 arg1 fluid 430:434 normal bovine synovial fluid 407:434 normal bovine synovial fluid 407:434 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 50 theme present 375:381 arg1 fragments 365:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 13 51 theme fragments 1574:1582 arg1 release 1554:1560 the release 1550:1560 the release of aggrecan fragments from articular cartilage into the synovial fluid 1550:1631 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 6 52 theme protein 877:883 arg1 cleavage 841:848 cleavage 841:848 cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains 841:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 13 53 theme articular 1589:1597 arg1 cartilage 1599:1607 articular cartilage 1589:1607 articular cartilage 1589:1607 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 16 54 theme cartilage 1770:1778 arg1 proteinase 1780:1789 a normal cartilage proteinase 1761:1789 a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain 1761:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 0 55 theme fragments 26:34 arg1 structure 4:12 The structure 0:12 The structure of aggrecan fragments in human synovial fluid 0:58 The structure of aggrecan fragments in human synovial fluid. 1569188 5 56 theme beginning 793:801 arg1 ARGSV 803:807 a single major sequence beginning ARGSV 769:807 a single major sequence beginning ARGSV 769:807 For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. 1569188 6 57 theme aggrecan 863:870 arg1 protein 877:883 the human aggrecan core protein 853:883 the human aggrecan core protein 853:883 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 16 58 theme 374 1821:1823 arg1 bond 1825:1828 the Glu 373-Ala 374 bond 1805:1828 the Glu 373-Ala 374 bond of the interglobular domain 1805:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 12 59 theme aggrecan 1237:1244 arg1 large 1348:1352 large 1348:1352 large 1348:1352 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 59 theme aggrecan 1237:1244 arg1 fragments 1246:1254 the major aggrecan fragments 1227:1254 the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid 1227:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 0 60 theme human 39:43 arg1 fluid 54:58 human synovial fluid 39:58 human synovial fluid 39:58 The structure of aggrecan fragments in human synovial fluid. 1569188 12 61 theme G2 1435:1436 arg1 domain 1438:1443 the G2 domain 1431:1443 the G2 domain 1431:1443 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 61 theme G2 1435:1436 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 62 theme domain 1466:1471 arg1 stretch 1394:1400 a short NH2-terminal stretch 1373:1400 a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s) 1373:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 63 theme chloride 461:468 arg1 centrifugation 479:492 cesium chloride gradient centrifugation 454:492 cesium chloride gradient centrifugation 454:492 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 64 theme gel 542:544 arg1 filtration 546:555 gel filtration 542:555 gel filtration on Superose-12 542:570 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 16 65 theme interglobular 1837:1849 arg1 domain 1851:1856 the interglobular domain 1833:1856 the interglobular domain 1833:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 4 66 theme core 614:617 arg1 populations 627:637 two major aggrecan core protein populations 595:637 two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD 595:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 6 67 from bond 908:911 arg1 cleavage 841:848 cleavage 841:848 cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains 841:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 1 68 from involvement 78:88 arg1 osteoarthritis 93:106 osteoarthritis 93:106 osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain 93:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 12 69 theme interglobular 1409:1421 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 69 theme interglobular 1409:1421 arg1 domain 1423:1428 the interglobular domain 1405:1428 the interglobular domain 1405:1428 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 16 70 theme Glu 1809:1811 arg1 bond 1825:1828 the Glu 373-Ala 374 bond 1805:1828 the Glu 373-Ala 374 bond of the interglobular domain 1805:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 6 71 theme human 857:861 arg1 protein 877:883 the human aggrecan core protein 853:883 the human aggrecan core protein 853:883 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 6 72 theme 373-Ala 896:902 arg1 bond 908:911 the Glu 373-Ala 374 bond 888:911 the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains 888:973 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 4 73 theme major 599:603 arg1 populations 627:637 two major aggrecan core protein populations 595:637 two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD 595:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 1 74 theme 373-Ala 152:158 arg1 bond 164:167 the Glu 373-Ala 374 bond 144:167 the Glu 373-Ala 374 bond of the interglobular domain 144:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 7 75 theme cartilage 1084:1092 arg1 cultures 1102:1109 bovine cartilage explant cultures 1077:1109 bovine cartilage explant cultures 1077:1109 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 4 76 theme kD 690:691 arg1 masses 663:668 apparent molecular masses 644:668 apparent molecular masses of approximately 90 kD and 150 kD 644:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 4 77 theme apparent 644:651 arg1 masses 663:668 apparent molecular masses 644:668 apparent molecular masses of approximately 90 kD and 150 kD 644:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 7 78 theme aggrecan 1054:1061 arg1 catabolism 1063:1072 interleukin-1 stimulated aggrecan catabolism 1029:1072 interleukin-1 stimulated aggrecan catabolism 1029:1072 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 1 79 theme domain 190:195 arg1 bond 164:167 the Glu 373-Ala 374 bond 144:167 the Glu 373-Ala 374 bond of the interglobular domain 144:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 2 80 with patients 232:239 arg1 injury 258:263 recent knee injury 246:263 recent knee injury 246:263 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 13 81 theme osteoarthritis 1661:1674 arg1 stages 1645:1650 all stages 1641:1650 all stages of human osteoarthritis 1641:1674 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 7 82 theme interleukin-1 1029:1041 arg1 catabolism 1063:1072 interleukin-1 stimulated aggrecan catabolism 1029:1072 interleukin-1 stimulated aggrecan catabolism 1029:1072 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 4 83 contain contained 585:593 arg2 populations 627:637 two major aggrecan core protein populations 595:637 two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD 595:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 4 83 contain contained 585:593 arg1 sample 578:583 Each sample 573:583 Each sample 573:583 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 12 84 theme bovine 1322:1327 arg1 fluid 1338:1342 normal bovine synovial fluid 1315:1342 normal bovine synovial fluid 1315:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 16 85 theme normal 1763:1768 arg1 proteinase 1780:1789 a normal cartilage proteinase 1761:1789 a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain 1761:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 2 86 theme recent 246:251 arg1 injury 258:263 recent knee injury 246:263 recent knee injury 246:263 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 1 87 theme novel 113:117 arg1 proteinase 119:128 a novel proteinase 111:128 a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain 111:195 Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 12 88 theme human 1287:1291 arg1 fluid 1302:1306 both osteoarthritic human synovial fluid 1267:1306 both osteoarthritic human synovial fluid 1267:1306 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 2 89 theme stage 302:306 arg1 osteoarthritis 308:321 early or late stage osteoarthritis 288:321 early or late stage osteoarthritis 288:321 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 7 90 from site 993:996 arg1 Cleavage 976:983 Cleavage 976:983 Cleavage at this site 976:996 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 12 91 theme sulfate 1514:1520 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 5 92 theme single 771:776 arg1 ARGSV 803:807 a single major sequence beginning ARGSV 769:807 a single major sequence beginning ARGSV 769:807 For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. 1569188 12 93 from fluid 1302:1306 arg1 present 1256:1262 present 1256:1262 present 1256:1262 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 0 94 from structure 4:12 arg1 fluid 54:58 human synovial fluid 39:58 human synovial fluid 39:58 The structure of aggrecan fragments in human synovial fluid. 1569188 3 95 from filtration 546:555 arg1 Superose-12 560:570 Superose-12 560:570 Superose-12 560:570 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 12 96 theme lengths 1487:1493 arg1 stretch 1394:1400 a short NH2-terminal stretch 1373:1400 a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s) 1373:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 97 theme gradient 470:477 arg1 centrifugation 479:492 cesium chloride gradient centrifugation 454:492 cesium chloride gradient centrifugation 454:492 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 3 98 theme sulfate-substituted 336:354 arg1 fragments 365:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments 324:373 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 7 99 dep occurs 1003:1008 arg1 P. 1123:1124 P. 1123:1124 P. 1123:1124 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 7 99 dep occurs 1003:1008 arg1 Flannery 1152:1159 Flannery 1152:1159 Flannery 1152:1159 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 7 99 dep occurs 1003:1008 arg1 J. 1119:1120 J. 1119:1120 J. 1119:1120 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 7 99 dep occurs 1003:1008 arg1 Neame 1126:1130 Neame 1126:1130 Neame 1126:1130 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 7 99 dep occurs 1003:1008 arg1 Boynton 1136:1142 Boynton 1136:1142 Boynton 1136:1142 Cleavage at this site also occurs during control and interleukin-1 stimulated aggrecan catabolism in bovine cartilage explant cultures (Sandy, J., P. Neame, R. Boynton, and C. Flannery. 1569188 3 100 from fluids 392:397 arg1 present 375:381 present 375:381 present 375:381 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 12 101 theme osteoarthritic 1272:1285 arg1 fluid 1302:1306 both osteoarthritic human synovial fluid 1267:1306 both osteoarthritic human synovial fluid 1267:1306 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 102 theme synovial 421:428 arg1 fluid 430:434 normal bovine synovial fluid 407:434 normal bovine synovial fluid 407:434 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 0 103 theme aggrecan 17:24 arg1 fragments 26:34 aggrecan fragments 17:34 aggrecan fragments 17:34 The structure of aggrecan fragments in human synovial fluid. 1569188 12 104 theme major 1231:1235 arg1 large 1348:1352 large 1348:1352 large 1348:1352 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 104 theme major 1231:1235 arg1 fragments 1246:1254 the major aggrecan fragments 1227:1254 the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid 1227:1342 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 105 theme cesium 454:459 arg1 centrifugation 479:492 cesium chloride gradient centrifugation 454:492 cesium chloride gradient centrifugation 454:492 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 5 106 theme sequence 784:791 arg1 ARGSV 803:807 a single major sequence beginning ARGSV 769:807 a single major sequence beginning ARGSV 769:807 For all samples, NH2-terminal analysis of both populations gave a single major sequence beginning ARGSV. 1569188 12 107 theme keratan 1450:1456 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 107 theme keratan 1450:1456 arg1 domain 1466:1471 the keratan sulfate domain 1446:1471 the keratan sulfate domain 1446:1471 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 6 108 theme core 872:875 arg1 protein 877:883 the human aggrecan core protein 853:883 the human aggrecan core protein 853:883 This NH2 terminus results from cleavage of the human aggrecan core protein at the Glu 373-Ala 374 bond within the interglobular domain between the G1 and G2 domains. 1569188 12 109 from fluid 1338:1342 arg1 present 1256:1262 present 1256:1262 present 1256:1262 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 110 theme normal 407:412 arg1 fluid 430:434 normal bovine synovial fluid 407:434 normal bovine synovial fluid 407:434 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 0 111 theme synovial 45:52 arg1 fluid 54:58 human synovial fluid 39:58 human synovial fluid 39:58 The structure of aggrecan fragments in human synovial fluid. 1569188 12 112 theme sulfate 1458:1464 arg1 s 1529:1529 the chondroitin sulfate domain(s) 1498:1530 the chondroitin sulfate domain(s) 1498:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 12 112 theme sulfate 1458:1464 arg1 domain 1466:1471 the keratan sulfate domain 1446:1471 the keratan sulfate domain 1446:1471 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 16 113 theme domain 1851:1856 arg1 bond 1825:1828 the Glu 373-Ala 374 bond 1805:1828 the Glu 373-Ala 374 bond of the interglobular domain 1805:1856 62:623-632) is promoted by the action of a normal cartilage proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. 1569188 4 114 theme protein 619:625 arg1 populations 627:637 two major aggrecan core protein populations 595:637 two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD 595:702 Each sample contained two major aggrecan core protein populations with apparent molecular masses of approximately 90 kD and 150 kD. 1569188 13 115 theme aggrecan 1565:1572 arg1 fragments 1574:1582 aggrecan fragments 1565:1582 aggrecan fragments from articular cartilage 1565:1607 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1569188 2 116 theme late 297:300 arg1 osteoarthritis 308:321 early or late stage osteoarthritis 288:321 early or late stage osteoarthritis 288:321 Synovial fluid was collected from patients with recent knee injury and from patients with early or late stage osteoarthritis. 1569188 12 117 theme domain 1438:1443 arg1 stretch 1394:1400 a short NH2-terminal stretch 1373:1400 a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s) 1373:1530 These results indicate that the major aggrecan fragments present in both osteoarthritic human synovial fluid and in normal bovine synovial fluid are large, being composed of a short NH2-terminal stretch of the interglobular domain, the G2 domain, the keratan sulfate domain, and variable lengths of the chondroitin sulfate domain(s). 1569188 3 118 from fluid 430:434 arg1 present 375:381 present 375:381 present 375:381 Chondroitin sulfate-substituted aggrecan fragments present in these fluids, and in normal bovine synovial fluid, were purified by cesium chloride gradient centrifugation, enzymically deglycosylated and fractionated by gel filtration on Superose-12. 1569188 13 119 from release 1554:1560 arg1 cartilage 1599:1607 articular cartilage 1589:1607 articular cartilage 1589:1607 We conclude that the release of aggrecan fragments from articular cartilage into the synovial fluid seen at all stages of human osteoarthritis (Lohmander, L. S. 1991. 1533633 0 0 theme lysosomal 78:86 arg1 enzyme 88:93 the lysosomal enzyme 74:93 the lysosomal enzyme 74:93 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 0 0 theme lysosomal 78:86 arg1 A 116:116 beta-hexosaminidase A 96:116 beta-hexosaminidase A 96:116 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 4 1 gly glycosylation 730:742 arg2 sites 744:748 the three glycosylation sites 720:748 the three glycosylation sites on the alpha-subunit 720:769 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 4 1 gly glycosylation 730:742 arg2 three 724:728 three 724:728 three 724:728 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 4 2 theme glycosylation 730:742 arg1 sites 744:748 the three glycosylation sites 720:748 the three glycosylation sites on the alpha-subunit 720:769 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 2 3 from sites 313:317 arg1 alpha-subunit 388:400 the alpha-subunit 384:400 the alpha-subunit of beta-hexosaminidase 384:423 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 9 4 theme disulfide 1463:1471 arg1 bonds 1473:1477 inappropriate disulfide bonds 1449:1477 inappropriate disulfide bonds 1449:1477 It was found to be improperly folded into an insoluble aggregate, linked through inappropriate disulfide bonds. 1533633 8 5 theme glycosylation 1250:1262 arg1 sites 1264:1268 all three glycosylation sites 1240:1268 all three glycosylation sites 1240:1268 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 11 6 theme Tay-Sachs 1743:1751 arg1 patients 1761:1768 Tay-Sachs disease patients 1743:1768 Tay-Sachs disease patients 1743:1768 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 10 7 gly unglycosylated 1484:1497 arg1 protein 1499:1505 The unglycosylated protein 1480:1505 The unglycosylated protein 1480:1505 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 1 8 theme lysosomal 271:279 arg1 enzymes 281:287 lysosomal enzymes 271:287 lysosomal enzymes 271:287 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 11 9 theme alpha-subunit 1683:1695 arg1 properties 1638:1647 The properties 1634:1647 The properties of the nonglycosylated, misfolded alpha-subunit 1634:1695 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 11 9 theme alpha-subunit 1683:1695 arg1 similar 1702:1708 similar 1702:1708 similar 1702:1708 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 10 10 with complex 1580:1586 arg1 BiP 1629:1631 BiP 1629:1631 BiP 1629:1631 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 10 10 with complex 1580:1586 arg1 protein 1620:1626 the Ig heavy chain-binding protein 1593:1626 the Ig heavy chain-binding protein 1593:1626 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 6 11 theme catalytic 981:989 arg1 activity 991:998 the catalytic activity 977:998 the catalytic activity associated with the alpha-chain 977:1030 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 1 12 theme enzymes 281:287 arg1 biosynthesis 255:266 the biosynthesis 251:266 the biosynthesis of lysosomal enzymes 251:287 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 13 dep glycosylation 153:165 arg1 The 149:151 The 149:151 The 149:151 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 11 14 theme mutant 1718:1723 arg1 alpha-subunits 1725:1738 some mutant alpha-subunits 1713:1738 some mutant alpha-subunits in Tay-Sachs disease patients 1713:1768 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 10 15 theme unglycosylated 1484:1497 arg1 protein 1499:1505 The unglycosylated protein 1480:1505 The unglycosylated protein 1480:1505 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 10 16 theme chain-binding 1606:1618 arg1 BiP 1629:1631 BiP 1629:1631 BiP 1629:1631 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 10 16 theme chain-binding 1606:1618 arg1 protein 1620:1626 the Ig heavy chain-binding protein 1593:1626 the Ig heavy chain-binding protein 1593:1626 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 6 17 theme oligosaccharide 926:940 arg1 absence 900:906 The absence 896:906 The absence of any individual oligosaccharide 896:940 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 8 18 gly glycosylation 1250:1262 arg2 sites 1264:1268 all three glycosylation sites 1240:1268 all three glycosylation sites 1240:1268 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 18 gly glycosylation 1250:1262 arg2 three 1244:1248 three 1244:1248 three 1244:1248 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 1 19 theme subsequent 171:180 arg1 modification 231:242 a pivotal modification 221:242 a pivotal modification during the biosynthesis of lysosomal enzymes 221:287 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 19 theme subsequent 171:180 arg1 glycosylation 153:165 glycosylation 153:165 glycosylation 153:165 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 19 theme subsequent 171:180 arg1 phosphorylation 182:196 subsequent phosphorylation 171:196 subsequent phosphorylation 171:196 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 0 20 theme beta-hexosaminidase 96:114 arg1 enzyme 88:93 the lysosomal enzyme 74:93 the lysosomal enzyme 74:93 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 0 20 theme beta-hexosaminidase 96:114 arg1 A 116:116 beta-hexosaminidase A 96:116 beta-hexosaminidase A 96:116 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 6 21 theme individual 915:924 arg1 oligosaccharide 926:940 any individual oligosaccharide 911:940 any individual oligosaccharide 911:940 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 2 22 theme enzyme 519:524 arg1 folding 490:496 folding 490:496 folding 490:496 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 22 theme enzyme 519:524 arg1 transport 502:510 transport 502:510 transport 502:510 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 11 23 gly nonglycosylated 1656:1670 arg1 alpha-subunit 1683:1695 the nonglycosylated, misfolded alpha-subunit 1652:1695 the nonglycosylated, misfolded alpha-subunit 1652:1695 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 4 24 theme COS-1 699:703 arg1 cells 705:709 COS-1 cells 699:709 COS-1 cells 699:709 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 11 25 from alpha-subunits 1725:1738 arg1 patients 1761:1768 Tay-Sachs disease patients 1743:1768 Tay-Sachs disease patients 1743:1768 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 12 26 theme proper 1877:1882 arg1 folding 1884:1890 proper folding 1877:1890 proper folding 1877:1890 The results indicate that the oligosaccharides are essential, although not in a site-specific manner, for proper folding and cellular transport of the alpha-subunit. 1533633 10 27 theme Ig 1597:1598 arg1 BiP 1629:1631 BiP 1629:1631 BiP 1629:1631 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 10 27 theme Ig 1597:1598 arg1 protein 1620:1626 the Ig heavy chain-binding protein 1593:1626 the Ig heavy chain-binding protein 1593:1626 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 2 28 theme glycosylation 331:343 arg1 phosphorylation 365:379 oligosaccharide phosphorylation 349:379 oligosaccharide phosphorylation 349:379 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 28 theme glycosylation 331:343 arg1 sites 313:317 the sites 309:317 the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase 309:423 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 28 theme glycosylation 331:343 arg1 glycosylation 331:343 N-linked glycosylation 322:343 N-linked glycosylation 322:343 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 5 29 theme oligosaccharides 836:851 arg1 oligosaccharides 836:851 the three oligosaccharides 826:851 the three oligosaccharides 826:851 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 29 theme oligosaccharides 836:851 arg1 phosphorylation 879:893 phosphorylation 879:893 phosphorylation 879:893 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 29 theme oligosaccharides 836:851 arg1 One 819:821 One 819:821 One 819:821 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 29 theme oligosaccharides 836:851 arg1 site 871:874 the preferred site 857:874 the preferred site of phosphorylation 857:893 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 8 30 theme wild-type 1290:1298 arg1 protein 1300:1306 the wild-type protein 1286:1306 the wild-type protein 1286:1306 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 4 31 mod modified 787:794 arg3 oligosaccharide 802:816 an oligosaccharide 799:816 an oligosaccharide 799:816 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 4 31 mod modified 787:794 arg1 sites 744:748 the three glycosylation sites 720:748 the three glycosylation sites on the alpha-subunit 720:769 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 4 31 mod modified 787:794 arg1 each 712:715 each 712:715 each 712:715 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 2 32 theme N-linked 322:329 arg1 glycosylation 331:343 N-linked glycosylation 322:343 N-linked glycosylation 322:343 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 1 33 theme mannose 201:207 arg1 residues 209:216 mannose residues 201:216 mannose residues 201:216 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 0 34 theme glycosylation 16:28 arg1 Analysis 0:7 Analysis 0:7 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 0:147 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 8 35 theme genetic 1218:1224 arg1 alteration 1226:1235 genetic alteration 1218:1235 genetic alteration of all three glycosylation sites 1218:1268 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 12 36 theme alpha-subunit 1922:1934 arg1 transport 1905:1913 cellular transport 1896:1913 cellular transport 1896:1913 The results indicate that the oligosaccharides are essential, although not in a site-specific manner, for proper folding and cellular transport of the alpha-subunit. 1533633 12 36 theme alpha-subunit 1922:1934 arg1 folding 1884:1890 proper folding 1877:1890 proper folding 1877:1890 The results indicate that the oligosaccharides are essential, although not in a site-specific manner, for proper folding and cellular transport of the alpha-subunit. 1533633 3 37 dep either 566:571 arg1 singly 573:578 singly 573:578 singly 573:578 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 1 38 theme residues 209:216 arg1 modification 231:242 a pivotal modification 221:242 a pivotal modification during the biosynthesis of lysosomal enzymes 221:287 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 38 theme residues 209:216 arg1 glycosylation 153:165 glycosylation 153:165 glycosylation 153:165 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 38 theme residues 209:216 arg1 phosphorylation 182:196 subsequent phosphorylation 171:196 subsequent phosphorylation 171:196 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 2 39 from influence 449:457 arg1 folding 490:496 folding 490:496 folding 490:496 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 39 from influence 449:457 arg1 transport 502:510 transport 502:510 transport 502:510 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 12 40 theme site-specific 1851:1863 arg1 manner 1865:1870 a site-specific manner 1849:1870 a site-specific manner 1849:1870 The results indicate that the oligosaccharides are essential, although not in a site-specific manner, for proper folding and cellular transport of the alpha-subunit. 1533633 10 41 theme reticulum 1551:1559 arg1 lumen 1526:1530 the lumen 1522:1530 the lumen of the endoplasmic reticulum 1522:1559 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 11 42 theme disease 1753:1759 arg1 patients 1761:1768 Tay-Sachs disease patients 1743:1768 Tay-Sachs disease patients 1743:1768 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 11 43 dep nonglycosylated 1656:1670 arg1 misfolded 1673:1681 misfolded 1673:1681 misfolded 1673:1681 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 0 44 theme phosphorylation 34:48 arg1 Analysis 0:7 Analysis 0:7 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 0:147 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 3 45 theme glycosylation 541:553 arg1 sequences 555:563 The potential glycosylation sequences 527:563 The potential glycosylation sequences 527:563 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 10 46 theme endoplasmic 1539:1549 arg1 reticulum 1551:1559 the endoplasmic reticulum 1535:1559 the endoplasmic reticulum 1535:1559 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 9 47 theme inappropriate 1449:1461 arg1 bonds 1473:1477 inappropriate disulfide bonds 1449:1477 inappropriate disulfide bonds 1449:1477 It was found to be improperly folded into an insoluble aggregate, linked through inappropriate disulfide bonds. 1533633 8 48 theme protein 1300:1306 arg1 synthesis 1273:1281 synthesis 1273:1281 synthesis of the wild-type protein 1273:1306 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 48 theme protein 1300:1306 arg1 alteration 1226:1235 genetic alteration 1218:1235 genetic alteration of all three glycosylation sites 1218:1268 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 49 theme tunicamycin 1327:1337 arg1 presence 1315:1322 the presence 1311:1322 the presence of tunicamycin 1311:1337 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 50 theme sites 1264:1268 arg1 synthesis 1273:1281 synthesis 1273:1281 synthesis of the wild-type protein 1273:1306 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 50 theme sites 1264:1268 arg1 alteration 1226:1235 genetic alteration 1218:1235 genetic alteration of all three glycosylation sites 1218:1268 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 3 51 dep sequences 555:563 arg1 combination 586:596 combination 586:596 combination 586:596 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 3 51 dep sequences 555:563 arg1 either 566:571 either 566:571 either 566:571 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 0 52 theme site-directed 122:134 arg1 mutagenesis 136:146 site-directed mutagenesis 122:146 site-directed mutagenesis 122:146 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 6 53 theme proper 1042:1047 arg1 folding 1049:1055 folding 1049:1055 folding 1049:1055 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 3 54 theme cDNA 656:659 arg1 mutagenesis 637:647 site-directed mutagenesis 623:647 site-directed mutagenesis of the cDNA 623:659 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 5 55 theme phosphorylation 879:893 arg1 oligosaccharides 836:851 the three oligosaccharides 826:851 the three oligosaccharides 826:851 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 55 theme phosphorylation 879:893 arg1 phosphorylation 879:893 phosphorylation 879:893 phosphorylation 879:893 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 55 theme phosphorylation 879:893 arg1 site 871:874 the preferred site 857:874 the preferred site of phosphorylation 857:893 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 55 theme phosphorylation 879:893 arg1 One 819:821 One 819:821 One 819:821 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 1 56 theme pivotal 223:229 arg1 modification 231:242 a pivotal modification 221:242 a pivotal modification during the biosynthesis of lysosomal enzymes 221:287 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 56 theme pivotal 223:229 arg1 glycosylation 153:165 glycosylation 153:165 glycosylation 153:165 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 1 56 theme pivotal 223:229 arg1 phosphorylation 182:196 subsequent phosphorylation 171:196 subsequent phosphorylation 171:196 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 6 57 theme activity 991:998 arg1 expression 963:972 the expression 959:972 the expression of the catalytic activity associated with the alpha-chain 959:1030 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 7 58 theme profound 1085:1092 arg1 effect 1094:1099 A profound effect 1083:1099 A profound effect 1083:1099 A profound effect was observed, however, when all three oligosaccharides were absent. 1533633 0 59 theme alpha-subunit 57:69 arg1 phosphorylation 34:48 phosphorylation 34:48 phosphorylation 34:48 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 0 59 theme alpha-subunit 57:69 arg1 glycosylation 16:28 glycosylation 16:28 glycosylation 16:28 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 12 60 theme cellular 1896:1903 arg1 transport 1905:1913 cellular transport 1896:1913 cellular transport 1896:1913 The results indicate that the oligosaccharides are essential, although not in a site-specific manner, for proper folding and cellular transport of the alpha-subunit. 1533633 8 61 gly unglycosylated 1173:1186 arg1 alpha-subunit 1188:1200 The unglycosylated alpha-subunit 1169:1200 The unglycosylated alpha-subunit 1169:1200 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 61 gly unglycosylated 1173:1186 arg1 inactive 1358:1365 inactive 1358:1365 inactive 1358:1365 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 3 62 theme potential 531:539 arg1 sequences 555:563 The potential glycosylation sequences 527:563 The potential glycosylation sequences 527:563 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 9 63 theme insoluble 1413:1421 arg1 aggregate 1423:1431 an insoluble aggregate 1410:1431 an insoluble aggregate 1410:1431 It was found to be improperly folded into an insoluble aggregate, linked through inappropriate disulfide bonds. 1533633 0 64 gly glycosylation 16:28 arg1 alpha-subunit 57:69 the alpha-subunit 53:69 the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, 53:117 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 1533633 2 65 theme phosphorylation 365:379 arg1 phosphorylation 365:379 oligosaccharide phosphorylation 349:379 oligosaccharide phosphorylation 349:379 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 65 theme phosphorylation 365:379 arg1 sites 313:317 the sites 309:317 the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase 309:423 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 65 theme phosphorylation 365:379 arg1 glycosylation 331:343 N-linked glycosylation 322:343 N-linked glycosylation 322:343 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 66 link N-linked 322:329 arg1 glycosylation 331:343 N-linked glycosylation 322:343 N-linked glycosylation 322:343 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 4 67 from expression 665:674 arg1 cells 705:709 COS-1 cells 699:709 COS-1 cells 699:709 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 2 68 theme oligosaccharide 349:363 arg1 phosphorylation 365:379 oligosaccharide phosphorylation 349:379 oligosaccharide phosphorylation 349:379 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 6 69 theme subunits 1073:1080 arg1 assembly 1061:1068 assembly 1061:1068 assembly 1061:1068 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 6 69 theme subunits 1073:1080 arg1 folding 1049:1055 folding 1049:1055 folding 1049:1055 The absence of any individual oligosaccharide did not diminish the expression of the catalytic activity associated with the alpha-chain, implying proper folding and assembly of subunits. 1533633 1 70 gly glycosylation 153:165 arg1 residues 209:216 mannose residues 201:216 mannose residues 201:216 The glycosylation and subsequent phosphorylation of mannose residues is a pivotal modification during the biosynthesis of lysosomal enzymes. 1533633 11 71 theme nonglycosylated 1656:1670 arg1 alpha-subunit 1683:1695 the nonglycosylated, misfolded alpha-subunit 1652:1695 the nonglycosylated, misfolded alpha-subunit 1652:1695 The properties of the nonglycosylated, misfolded alpha-subunit were similar to some mutant alpha-subunits in Tay-Sachs disease patients. 1533633 10 72 located found 1569:1573 arg1 complex 1580:1586 a complex 1578:1586 a complex with the Ig heavy chain-binding protein, BiP 1578:1631 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 10 72 located found 1569:1573 arg2 protein 1499:1505 The unglycosylated protein 1480:1505 The unglycosylated protein 1480:1505 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 8 73 theme unglycosylated 1173:1186 arg1 alpha-subunit 1188:1200 The unglycosylated alpha-subunit 1169:1200 The unglycosylated alpha-subunit 1169:1200 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 8 73 theme unglycosylated 1173:1186 arg1 inactive 1358:1365 inactive 1358:1365 inactive 1358:1365 The unglycosylated alpha-subunit, resulting from genetic alteration of all three glycosylation sites or synthesis of the wild-type protein in the presence of tunicamycin, was catalytically inactive. 1533633 4 74 theme cDNAs 690:694 arg1 expression 665:674 expression 665:674 expression of the mutant cDNAs in COS-1 cells 665:709 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 2 75 theme oligosaccharides 466:481 arg1 influence 449:457 the influence 445:457 the influence of the oligosaccharides on the folding and transport of the enzyme 445:524 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 76 theme beta-hexosaminidase 405:423 arg1 alpha-subunit 388:400 the alpha-subunit 384:400 the alpha-subunit of beta-hexosaminidase 384:423 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 4 77 from sites 744:748 arg1 alpha-subunit 757:769 the alpha-subunit 753:769 the alpha-subunit 753:769 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 2 78 gly glycosylation 331:343 arg2 glycosylation 331:343 N-linked glycosylation 322:343 N-linked glycosylation 322:343 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 78 gly glycosylation 331:343 arg2 sites 313:317 the sites 309:317 the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase 309:423 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 2 78 gly glycosylation 331:343 arg2 phosphorylation 365:379 oligosaccharide phosphorylation 349:379 oligosaccharide phosphorylation 349:379 We have identified the sites of N-linked glycosylation and oligosaccharide phosphorylation on the alpha-subunit of beta-hexosaminidase and have determined the influence of the oligosaccharides on the folding and transport of the enzyme. 1533633 4 79 theme mutant 683:688 arg1 cDNAs 690:694 the mutant cDNAs 679:694 the mutant cDNAs 679:694 By expression of the mutant cDNAs in COS-1 cells, each of the three glycosylation sites on the alpha-subunit was found to be modified by an oligosaccharide. 1533633 10 80 theme heavy 1600:1604 arg1 BiP 1629:1631 BiP 1629:1631 BiP 1629:1631 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 10 80 theme heavy 1600:1604 arg1 protein 1620:1626 the Ig heavy chain-binding protein 1593:1626 the Ig heavy chain-binding protein 1593:1626 The unglycosylated protein was trapped in the lumen of the endoplasmic reticulum and was found in a complex with the Ig heavy chain-binding protein, BiP. 1533633 3 81 theme site-directed 623:635 arg1 mutagenesis 637:647 site-directed mutagenesis 623:647 site-directed mutagenesis of the cDNA 623:659 The potential glycosylation sequences, either singly or in combination, were eliminated through site-directed mutagenesis of the cDNA. 1533633 5 82 theme preferred 861:869 arg1 oligosaccharides 836:851 the three oligosaccharides 826:851 the three oligosaccharides 826:851 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 82 theme preferred 861:869 arg1 phosphorylation 879:893 phosphorylation 879:893 phosphorylation 879:893 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 82 theme preferred 861:869 arg1 site 871:874 the preferred site 857:874 the preferred site of phosphorylation 857:893 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 5 82 theme preferred 861:869 arg1 One 819:821 One 819:821 One 819:821 One of the three oligosaccharides was the preferred site of phosphorylation. 1533633 0 83 theme enzyme 88:93 arg1 alpha-subunit 57:69 the alpha-subunit 53:69 the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, 53:117 Analysis of the glycosylation and phosphorylation of the alpha-subunit of the lysosomal enzyme, beta-hexosaminidase A, by site-directed mutagenesis. 20457942 1 0 theme cell 92:95 arg1 membranes 97:105 cell membranes 92:105 cell membranes 92:105 In humans, NH(3) transport across cell membranes is facilitated by the Rh (rhesus) family of proteins. 20457942 5 1 theme RhCG 637:640 arg1 structure 642:650 our RhCG structure 633:650 our RhCG structure 633:650 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 2 theme RhCE 765:768 arg1 heterotrimers 733:745 stochastically assembled heterotrimers 708:745 stochastically assembled heterotrimers of RhAG, RhD, and RhCE 708:768 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 3 theme erythrocyte 601:611 arg1 complex 616:622 the erythrocyte Rh complex 597:622 the erythrocyte Rh complex 597:622 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 4 4 contain contains 433:440 arg1 monomer 425:431 Each monomer 420:431 Each monomer 420:431 Each monomer contains 12 transmembrane helices, one more than in the bacterial homologs. 20457942 4 4 contain contains 433:440 arg2 one 468:470 one 468:470 one 468:470 Each monomer contains 12 transmembrane helices, one more than in the bacterial homologs. 20457942 4 4 contain contains 433:440 arg2 helices 459:465 12 transmembrane helices 442:465 12 transmembrane helices 442:465 Each monomer contains 12 transmembrane helices, one more than in the bacterial homologs. 20457942 2 5 theme ammonia 249:255 arg1 excretion 257:265 ammonia excretion 249:265 ammonia excretion 249:265 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 5 6 theme Rh 613:614 arg1 complex 616:622 the erythrocyte Rh complex 597:622 the erythrocyte Rh complex 597:622 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 7 theme RhAG 750:753 arg1 heterotrimers 733:745 stochastically assembled heterotrimers 708:745 stochastically assembled heterotrimers of RhAG, RhD, and RhCE 708:768 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 3 8 theme RhCG 338:341 arg1 structure 319:327 The X-ray crystallographic structure 292:327 The X-ray crystallographic structure 292:327 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 20457942 5 9 theme complex 616:622 arg1 Models 587:592 internal pH. Models 574:592 internal pH. Models of the erythrocyte Rh complex based on our RhCG structure 574:650 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 10 theme Rh 682:683 arg1 complex 685:691 the erythrocytic Rh complex 665:691 the erythrocytic Rh complex 665:691 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 11 theme erythrocytic 669:680 arg1 complex 685:691 the erythrocytic Rh complex 665:691 the erythrocytic Rh complex 665:691 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 2 12 theme pH 277:278 arg1 regulation 280:289 renal pH regulation 271:289 renal pH regulation 271:289 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 2 13 gly glycoprotein 172:183 arg1 RhCG 186:189 RhCG 186:189 RhCG 186:189 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 2 13 gly glycoprotein 172:183 arg1 glycoprotein 172:183 Human Rh C glycoprotein 161:183 Human Rh C glycoprotein (RhCG) 161:190 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 4 14 theme transmembrane 445:457 arg1 one 468:470 one 468:470 one 468:470 Each monomer contains 12 transmembrane helices, one more than in the bacterial homologs. 20457942 4 14 theme transmembrane 445:457 arg1 helices 459:465 12 transmembrane helices 442:465 12 transmembrane helices 442:465 Each monomer contains 12 transmembrane helices, one more than in the bacterial homologs. 20457942 2 15 theme renal 271:275 arg1 regulation 280:289 renal pH regulation 271:289 renal pH regulation 271:289 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 0 16 theme Rh 18:19 arg1 Function 0:7 Function 0:7 Function of human Rh 0:19 Function of human Rh based on structure of RhCG at 2.1 A. 20457942 5 17 theme internal 574:581 arg1 Models 587:592 internal pH. Models 574:592 internal pH. Models of the erythrocyte Rh complex based on our RhCG structure 574:650 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 2 18 theme trimeric 200:207 arg1 complex 209:215 a trimeric complex 198:215 a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation 198:289 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 1 19 theme Rh 129:130 arg1 family 141:146 the Rh (rhesus) family 125:146 the Rh (rhesus) family of proteins 125:158 In humans, NH(3) transport across cell membranes is facilitated by the Rh (rhesus) family of proteins. 20457942 0 20 theme human 12:16 arg1 Rh 18:19 human Rh 12:19 human Rh 12:19 Function of human Rh based on structure of RhCG at 2.1 A. 20457942 5 21 theme pH. 583:585 arg1 Models 587:592 internal pH. Models 574:592 internal pH. Models of the erythrocyte Rh complex based on our RhCG structure 574:650 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 0 22 from A 55:55 arg1 structure 30:38 structure 30:38 structure of RhCG at 2.1 A 30:55 Function of human Rh based on structure of RhCG at 2.1 A. 20457942 3 23 theme A 362:362 arg1 resolution 364:373 2.1 A resolution 358:373 2.1 A resolution 358:373 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 20457942 3 24 theme crystallographic 302:317 arg1 structure 319:327 The X-ray crystallographic structure 292:327 The X-ray crystallographic structure 292:327 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 20457942 3 25 theme human 332:336 arg1 RhCG 338:341 human RhCG 332:341 human RhCG 332:341 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 20457942 1 26 theme rhesus 133:138 arg1 family 141:146 the Rh (rhesus) family 125:146 the Rh (rhesus) family of proteins 125:158 In humans, NH(3) transport across cell membranes is facilitated by the Rh (rhesus) family of proteins. 20457942 5 27 dep RhCG 545:548 arg1 conducts 550:557 conducts 550:557 conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure 550:650 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 28 theme assembled 723:731 arg1 heterotrimers 733:745 stochastically assembled heterotrimers 708:745 stochastically assembled heterotrimers of RhAG, RhD, and RhCE 708:768 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 5 29 theme RhD 756:758 arg1 heterotrimers 733:745 stochastically assembled heterotrimers 708:745 stochastically assembled heterotrimers of RhAG, RhD, and RhCE 708:768 Reconstituted into proteoliposomes, RhCG conducts NH(3) to raise internal pH. Models of the erythrocyte Rh complex based on our RhCG structure suggest that the erythrocytic Rh complex is composed of stochastically assembled heterotrimers of RhAG, RhD, and RhCE. 20457942 3 30 theme X-ray 296:300 arg1 structure 319:327 The X-ray crystallographic structure 292:327 The X-ray crystallographic structure 292:327 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 20457942 2 31 theme essential 231:239 arg1 role 241:244 an essential role 228:244 an essential role 228:244 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 1 32 theme 3 72:72 arg1 transport 75:83 NH(3) transport 69:83 NH(3) transport across cell membranes 69:105 In humans, NH(3) transport across cell membranes is facilitated by the Rh (rhesus) family of proteins. 20457942 4 33 theme bacterial 489:497 arg1 homologs 499:506 the bacterial homologs 485:506 the bacterial homologs 485:506 Each monomer contains 12 transmembrane helices, one more than in the bacterial homologs. 20457942 2 34 theme C 170:170 arg1 RhCG 186:189 RhCG 186:189 RhCG 186:189 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 2 34 theme C 170:170 arg1 glycoprotein 172:183 Human Rh C glycoprotein 161:183 Human Rh C glycoprotein (RhCG) 161:190 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 2 35 theme Rh 167:168 arg1 RhCG 186:189 RhCG 186:189 RhCG 186:189 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 2 35 theme Rh 167:168 arg1 glycoprotein 172:183 Human Rh C glycoprotein 161:183 Human Rh C glycoprotein (RhCG) 161:190 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 3 36 theme ammonia 401:407 arg1 transport 409:417 ammonia transport 401:417 ammonia transport 401:417 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 20457942 1 37 theme proteins 151:158 arg1 family 141:146 the Rh (rhesus) family 125:146 the Rh (rhesus) family of proteins 125:158 In humans, NH(3) transport across cell membranes is facilitated by the Rh (rhesus) family of proteins. 20457942 0 38 theme RhCG 43:46 arg1 structure 30:38 structure 30:38 structure of RhCG at 2.1 A 30:55 Function of human Rh based on structure of RhCG at 2.1 A. 20457942 2 39 theme Human 161:165 arg1 RhCG 186:189 RhCG 186:189 RhCG 186:189 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 2 39 theme Human 161:165 arg1 glycoprotein 172:183 Human Rh C glycoprotein 161:183 Human Rh C glycoprotein (RhCG) 161:190 Human Rh C glycoprotein (RhCG) forms a trimeric complex that plays an essential role in ammonia excretion and renal pH regulation. 20457942 3 40 theme transport 409:417 arg1 mechanism 388:396 the mechanism 384:396 the mechanism of ammonia transport 384:417 The X-ray crystallographic structure of human RhCG, determined at 2.1 A resolution, reveals the mechanism of ammonia transport. 3497398 3 0 with treatment 516:524 arg1 bromide 540:546 cyanogen bromide 531:546 cyanogen bromide 531:546 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 3 0 with treatment 516:524 arg1 protease 566:573 Achromobacter protease I 552:575 Achromobacter protease I 552:575 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 2 1 theme von 336:338 arg1 domain 366:371 the von Willebrand factor-binding domain 332:371 the von Willebrand factor-binding domain 332:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 8 2 theme charged 1208:1214 arg1 molecules 1216:1224 positively charged molecules 1197:1224 positively charged molecules 1197:1224 This area is likely to attract positively charged molecules. 3497398 9 3 dep region 1299:1304 arg1 corresponding 1306:1318 corresponding 1306:1318 corresponding to the previously described "macroglycopeptide" 1306:1366 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 9 3 dep region 1299:1304 arg1 representing 1372:1383 representing 1372:1383 representing the carboxyl-terminal half of the alpha chain 1372:1429 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 4 4 theme aureus 692:697 arg1 protease 702:709 Staphylococcus aureus V8 protease 677:709 Staphylococcus aureus V8 protease 677:709 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 11 5 theme protein 1698:1704 arg1 sequence 1706:1713 This protein sequence 1693:1713 This protein sequence 1693:1713 This protein sequence agrees completely with that deduced from the cDNA sequence reported by Lopez et al. [Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T. & Roth, G.J. (1987) Proc. 3497398 10 6 theme determined 1470:1479 arg1 sequence 1481:1488 The determined sequence 1466:1488 The determined sequence of the alpha chain of glycoprotein Ib 1466:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 2 7 theme complete 248:255 arg1 sequence 257:264 the complete sequence 244:264 the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain 244:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 6 8 theme glycoprotein 929:940 arg1 Ib 942:943 glycoprotein Ib 929:943 glycoprotein Ib 929:943 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 6 9 contain contains 972:979 arg2 chains 1029:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 6 9 contain contains 972:979 arg1 region 919:924 This region 914:924 This region of glycoprotein Ib 914:943 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 7 10 theme charged 1137:1143 arg1 residues 1145:1152 10 negatively charged residues 1123:1152 10 negatively charged residues 1123:1152 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 2 11 theme fragment 296:303 arg1 sequence 257:264 the complete sequence 244:264 the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain 244:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 1 12 theme alpha 170:174 arg1 chain 176:180 the alpha chain 166:180 the alpha chain of the human platelet membrane glycoprotein Ib 166:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 10 13 theme duplication 1612:1622 arg1 region 1539:1544 a region 1537:1544 a region (residues 29-193) 1537:1562 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 13 theme duplication 1612:1622 arg1 indicative 1593:1602 indicative 1593:1602 indicative 1593:1602 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 13 theme duplication 1612:1622 arg1 homologous 1638:1647 homologous 1638:1647 homologous 1638:1647 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 2 14 theme amino-terminal 273:286 arg1 fragment 296:303 the amino-terminal tryptic fragment 269:303 the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain 269:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 6 15 theme O-linked 1007:1014 arg1 chains 1029:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 10 16 theme human 1652:1656 arg1 2-glycoprotein 1677:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 6 17 link O-linked 1007:1014 arg1 chains 1029:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 5 18 theme polypeptide 901:911 arg1 half 877:880 approximately half 863:880 approximately half of the alpha-chain polypeptide 863:911 Analysis of all these fragments provided data that allowed determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide. 3497398 1 19 from chain 176:180 arg1 segment 153:159 a 299-residue segment 139:159 a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib 139:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 1 19 from chain 176:180 arg1 sequence 127:134 the amino acid sequence 112:134 the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib 112:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 9 20 gly macroglycopeptide 1349:1365 arg2 macroglycopeptide 1349:1365 the previously described "macroglycopeptide" 1323:1366 the previously described "macroglycopeptide" 1323:1366 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 1 21 theme acid 122:125 arg1 sequence 127:134 the amino acid sequence 112:134 the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib 112:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 0 22 theme domain 57:62 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 0:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 10 23 theme alpha 1497:1501 arg1 chain 1503:1507 the alpha chain 1493:1507 the alpha chain of glycoprotein Ib 1493:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 0 24 theme platelet 67:74 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 1 25 theme 299-residue 141:151 arg1 segment 153:159 a 299-residue segment 139:159 a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib 139:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 9 26 dep glycosylated 1251:1262 arg1 threonine 1279:1287 threonine 1279:1287 threonine 1279:1287 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 9 26 dep glycosylated 1251:1262 arg1 serine 1268:1273 serine 1268:1273 serine 1268:1273 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 0 27 theme glycoprotein 85:96 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 1 28 from sequence 127:134 arg1 chain 176:180 the alpha chain 166:180 the alpha chain of the human platelet membrane glycoprotein Ib 166:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 4 29 dep Serratia 716:723 arg1 marcescens 725:734 marcescens 725:734 marcescens 725:734 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 10 30 theme Ib 1525:1526 arg1 chain 1503:1507 the alpha chain 1493:1507 the alpha chain of glycoprotein Ib 1493:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 3 31 theme primary 378:384 arg1 sets 386:389 Two primary sets 374:389 Two primary sets of overlapping fragments 374:414 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 3 31 theme primary 378:384 arg1 fragments 406:414 overlapping fragments 394:414 overlapping fragments 394:414 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 0 32 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 0:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 9 33 theme carboxyl-terminal 1389:1405 arg1 half 1407:1410 the carboxyl-terminal half 1385:1410 the carboxyl-terminal half of the alpha chain 1385:1429 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 3 34 theme protein 469:475 arg1 cleavage 433:440 cleavage 433:440 cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds 433:504 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 3 35 theme fragments 406:414 arg1 sets 386:389 Two primary sets 374:389 Two primary sets of overlapping fragments 374:414 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 3 35 theme fragments 406:414 arg1 fragments 406:414 overlapping fragments 394:414 overlapping fragments 394:414 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 10 36 with region 1539:1544 arg1 repeats 1575:1581 seven repeats 1569:1581 seven repeats 1569:1581 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 9 37 theme alpha 1419:1423 arg1 chain 1425:1429 the alpha chain 1415:1429 the alpha chain 1415:1429 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 10 38 dep residues 1547:1554 arg1 29-193 1556:1561 29-193 1556:1561 29-193 1556:1561 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 3 39 theme lysyl 494:498 arg1 bonds 500:504 methionyl and lysyl bonds 480:504 methionyl and lysyl bonds 480:504 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 5 40 theme sequence 837:844 arg1 determination 805:817 determination 805:817 determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide 805:911 Analysis of all these fragments provided data that allowed determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide. 3497398 10 41 gly glycoprotein 1512:1523 arg1 glycoprotein 1512:1523 glycoprotein Ib 1512:1526 glycoprotein Ib 1512:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 2 42 theme residues 312:319 arg1 fragment 296:303 the amino-terminal tryptic fragment 269:303 the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain 269:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 1 43 theme platelet 195:202 arg1 glycoprotein 213:224 the human platelet membrane glycoprotein 185:224 the human platelet membrane glycoprotein Ib 185:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 11 44 theme cDNA 1760:1763 arg1 sequence 1765:1772 the cDNA sequence 1756:1772 the cDNA sequence reported by Lopez et al. [Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T. & Roth, G.J. (1987) Proc 1756:1896 This protein sequence agrees completely with that deduced from the cDNA sequence reported by Lopez et al. [Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T. & Roth, G.J. (1987) Proc. 3497398 1 45 theme glycoprotein 213:224 arg1 Ib 226:227 the human platelet membrane glycoprotein Ib 185:227 the human platelet membrane glycoprotein Ib 185:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 5 46 theme continuous 826:835 arg1 sequence 837:844 the continuous sequence 822:844 the continuous sequence corresponding to approximately half of the alpha-chain polypeptide 822:911 Analysis of all these fragments provided data that allowed determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide. 3497398 2 47 theme Willebrand 340:349 arg1 domain 366:371 the von Willebrand factor-binding domain 332:371 the von Willebrand factor-binding domain 332:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 4 48 theme Additional 592:601 arg1 fragments 603:611 Additional fragments 592:611 Additional fragments 592:611 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 10 49 theme gene 1607:1610 arg1 duplication 1612:1622 gene duplication 1607:1622 gene duplication 1607:1622 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 9 50 theme hydrophilic 1231:1241 arg1 region 1299:1304 The hydrophilic, highly glycosylated (at serine and threonine residues) region 1227:1304 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain 1227:1429 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 9 50 theme hydrophilic 1231:1241 arg1 likely 1434:1439 likely 1434:1439 likely 1434:1439 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 3 51 theme Achromobacter 552:564 arg1 protease 566:573 Achromobacter protease I 552:575 Achromobacter protease I 552:575 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 7 52 theme residues 1145:1152 arg1 cluster 1112:1118 a cluster 1110:1118 a cluster of 10 negatively charged residues at 269-287 1110:1163 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 1 53 theme segment 153:159 arg1 sequence 127:134 the amino acid sequence 112:134 the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib 112:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 0 54 theme Willebrand 31:40 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 0 54 theme Willebrand 31:40 arg1 domain 57:62 the von Willebrand factor-binding domain 23:62 the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 23:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 6 55 theme Ib 942:943 arg1 region 919:924 This region 914:924 This region of glycoprotein Ib 914:943 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 4 56 theme Staphylococcus 677:690 arg1 protease 702:709 Staphylococcus aureus V8 protease 677:709 Staphylococcus aureus V8 protease 677:709 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 4 57 theme V8 699:700 arg1 protease 702:709 Staphylococcus aureus V8 protease 677:709 Staphylococcus aureus V8 protease 677:709 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 3 58 from bonds 500:504 arg1 cleavage 433:440 cleavage 433:440 cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds 433:504 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 2 59 theme tryptic 288:294 arg1 fragment 296:303 the amino-terminal tryptic fragment 269:303 the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain 269:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 6 60 theme carbohydrate 1016:1027 arg1 chains 1029:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 4 61 theme glycocalicin 650:661 arg1 treatment 630:638 treatment 630:638 treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease 630:743 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 10 62 theme leucine-rich 1658:1669 arg1 2-glycoprotein 1677:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 11 63 dep Lopez 1786:1790 arg1 al. 1795:1797 Lopez et al. 1786:1797 Lopez et al. [Lopez 1786:1804 This protein sequence agrees completely with that deduced from the cDNA sequence reported by Lopez et al. [Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T. & Roth, G.J. (1987) Proc. 3497398 5 64 theme alpha-chain 889:899 arg1 polypeptide 901:911 the alpha-chain polypeptide 885:911 the alpha-chain polypeptide 885:911 Analysis of all these fragments provided data that allowed determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide. 3497398 4 65 with treatment 630:638 arg1 trypsin 668:674 trypsin 668:674 trypsin 668:674 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 4 65 with treatment 630:638 arg1 protease 736:743 Serratia marcescens protease 716:743 Serratia marcescens protease 716:743 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 4 65 with treatment 630:638 arg1 protease 702:709 Staphylococcus aureus V8 protease 677:709 Staphylococcus aureus V8 protease 677:709 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 10 66 contain contains 1528:1535 arg2 indicative 1593:1602 indicative 1593:1602 indicative 1593:1602 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 66 contain contains 1528:1535 arg1 sequence 1481:1488 The determined sequence 1466:1488 The determined sequence of the alpha chain of glycoprotein Ib 1466:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 66 contain contains 1528:1535 arg2 homologous 1638:1647 homologous 1638:1647 homologous 1638:1647 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 66 contain contains 1528:1535 arg2 residues 1547:1554 residues 29-193 1547:1561 residues 29-193 1547:1561 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 66 contain contains 1528:1535 arg2 region 1539:1544 a region 1537:1544 a region (residues 29-193) 1537:1562 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 6 67 theme N-linked 990:997 arg1 chains 1029:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 6 68 link N-linked 990:997 arg1 chains 1029:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 only two N-linked and one O-linked carbohydrate chains 981:1034 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 1 69 theme amino 116:120 arg1 sequence 127:134 the amino acid sequence 112:134 the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib 112:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 6 70 gly glycoprotein 929:940 arg1 glycoprotein 929:940 glycoprotein Ib 929:943 glycoprotein Ib 929:943 This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains. 3497398 3 71 theme methionyl 480:488 arg1 bonds 500:504 methionyl and lysyl bonds 480:504 methionyl and lysyl bonds 480:504 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 10 72 gly 2-glycoprotein 1677:1690 arg1 2-glycoprotein 1677:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 9 73 dep hydrophilic 1231:1241 arg1 glycosylated 1251:1262 glycosylated 1251:1262 glycosylated 1251:1262 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 11 74 theme Lopez 1786:1790 arg1 D.W. 1820:1823 D.W. 1820:1823 D.W. 1820:1823 This protein sequence agrees completely with that deduced from the cDNA sequence reported by Lopez et al. [Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T. & Roth, G.J. (1987) Proc. 3497398 11 74 theme Lopez 1786:1790 arg1 [Lopez 1799:1804 Lopez et al. [Lopez 1786:1804 Lopez et al. [Lopez 1786:1804 This protein sequence agrees completely with that deduced from the cDNA sequence reported by Lopez et al. [Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T. & Roth, G.J. (1987) Proc. 3497398 0 75 theme membrane 76:83 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 9 76 theme described 1338:1346 arg1 " 1366:1366 the previously described "macroglycopeptide" 1323:1366 the previously described "macroglycopeptide" 1323:1366 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 7 77 theme hydrophilic 1039:1049 arg1 region 1051:1056 A hydrophilic region 1037:1056 A hydrophilic region 1037:1056 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 9 78 dep serine 1268:1273 arg1 residues 1289:1296 residues 1289:1296 residues 1289:1296 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 4 79 theme Serratia 716:723 arg1 protease 736:743 Serratia marcescens protease 716:743 Serratia marcescens protease 716:743 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 0 80 theme Ib 98:99 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 0 80 theme Ib 98:99 arg1 domain 57:62 the von Willebrand factor-binding domain 23:62 the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 23:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 9 81 theme macroglycopeptide 1349:1365 arg1 " 1366:1366 the previously described "macroglycopeptide" 1323:1366 the previously described "macroglycopeptide" 1323:1366 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 5 82 theme fragments 768:776 arg1 Analysis 746:753 Analysis 746:753 Analysis of all these fragments 746:776 Analysis of all these fragments provided data that allowed determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide. 3497398 1 83 gly glycoprotein 213:224 arg1 glycoprotein 213:224 the human platelet membrane glycoprotein 185:224 the human platelet membrane glycoprotein Ib 185:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 10 84 theme glycoprotein 1512:1523 arg1 Ib 1525:1526 glycoprotein Ib 1512:1526 glycoprotein Ib 1512:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 7 85 contain contains 1101:1108 arg1 residues 1073:1080 residues 215 and 299 1073:1092 residues 1073:1080 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 7 85 contain contains 1101:1108 arg2 cluster 1112:1118 a cluster 1110:1118 a cluster of 10 negatively charged residues at 269-287 1110:1163 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 7 85 contain contains 1101:1108 arg1 299 1090:1092 299 1090:1092 299 1090:1092 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 0 86 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 0:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 0 87 gly glycoprotein 85:96 arg1 glycoprotein 85:96 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 10 88 theme chain 1503:1507 arg1 sequence 1481:1488 The determined sequence 1466:1488 The determined sequence of the alpha chain of glycoprotein Ib 1466:1526 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 10 89 theme alpha 1671:1675 arg1 2-glycoprotein 1677:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 human leucine-rich alpha 2-glycoprotein 1652:1690 The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein. 3497398 0 90 theme von 27:29 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 0 90 theme von 27:29 arg1 domain 57:62 the von Willebrand factor-binding domain 23:62 the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 23:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 7 91 from 269-287 1157:1163 arg1 cluster 1112:1118 a cluster 1110:1118 a cluster of 10 negatively charged residues at 269-287 1110:1163 A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287. 3497398 3 92 theme S-carboxymethylated 449:467 arg1 protein 469:475 the S-carboxymethylated protein 445:475 the S-carboxymethylated protein 445:475 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 0 93 theme factor-binding 42:55 arg1 Ib 98:99 platelet membrane glycoprotein Ib 67:99 platelet membrane glycoprotein Ib 67:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 0 93 theme factor-binding 42:55 arg1 domain 57:62 the von Willebrand factor-binding domain 23:62 the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib 23:99 Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. 3497398 9 94 theme chain 1425:1429 arg1 half 1407:1410 the carboxyl-terminal half 1385:1410 the carboxyl-terminal half of the alpha chain 1385:1429 The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292. 3497398 3 95 theme overlapping 394:404 arg1 fragments 406:414 overlapping fragments 394:414 overlapping fragments 394:414 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 2 96 theme factor-binding 351:364 arg1 domain 366:371 the von Willebrand factor-binding domain 332:371 the von Willebrand factor-binding domain 332:371 This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain. 3497398 1 97 theme membrane 204:211 arg1 glycoprotein 213:224 the human platelet membrane glycoprotein 185:224 the human platelet membrane glycoprotein Ib 185:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 1 98 theme human 189:193 arg1 glycoprotein 213:224 the human platelet membrane glycoprotein 185:224 the human platelet membrane glycoprotein Ib 185:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 3497398 4 99 theme native 643:648 arg1 glycocalicin 650:661 native glycocalicin 643:661 native glycocalicin 643:661 Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease. 3497398 3 100 theme cyanogen 531:538 arg1 bromide 540:546 cyanogen bromide 531:546 cyanogen bromide 531:546 Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively. 3497398 1 101 theme Ib 226:227 arg1 chain 176:180 the alpha chain 166:180 the alpha chain of the human platelet membrane glycoprotein Ib 166:227 We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib. 19855092 10 0 from improvements 1816:1827 arg1 tests 1862:1866 glucose and insulin tolerance tests 1832:1866 glucose and insulin tolerance tests observed with control adiponectin 1832:1900 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 6 1 theme adiponectin 915:925 arg1 forms 906:910 mutant forms 899:910 mutant forms of adiponectin (lacking the modified Thr residues) 899:961 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 2 2 theme proline 380:386 arg1 hydroxylation 388:400 proline hydroxylation 380:400 proline hydroxylation 380:400 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 9 3 theme rat 1537:1539 arg1 hepatocytes 1541:1551 isolated primary rat hepatocytes 1520:1551 isolated primary rat hepatocytes 1520:1551 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 4 4 theme unidentified 678:689 arg1 glycans 700:706 previously unidentified O-linked glycans 667:706 previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin 667:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 6 5 theme forms 906:910 arg1 Expression 885:894 Expression 885:894 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation 885:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 4 6 from adiponectin 756:766 arg1 residues 715:722 Thr residues 711:722 Thr residues of the variable domain in human adiponectin 711:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 4 6 from adiponectin 756:766 arg1 domain 740:745 the variable domain 727:745 the variable domain in human adiponectin 727:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 6 7 theme adiponectin 979:989 arg1 Expression 885:894 Expression 885:894 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation 885:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 6 8 from adiponectin 979:989 arg1 cells 994:998 cells 994:998 cells defective in sialylation 994:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 9 9 theme primary 1529:1535 arg1 hepatocytes 1541:1551 isolated primary rat hepatocytes 1520:1551 isolated primary rat hepatocytes 1520:1551 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 2 10 theme Previous 290:297 arg1 work 299:302 Previous work 290:302 Previous work 290:302 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 5 11 theme Enzymatic 769:777 arg1 removal 779:785 Enzymatic removal 769:785 Enzymatic removal of sialic acid or its underlying O-linked sugars 769:834 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 10 12 theme desialylated 1724:1735 arg1 adiponectin 1737:1747 desialylated adiponectin 1724:1747 desialylated adiponectin 1724:1747 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 11 13 theme acid 1951:1954 arg1 content 1956:1962 sialic acid content 1944:1962 sialic acid content 1944:1962 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 8 14 theme control 1363:1369 arg1 adiponectin 1371:1381 control adiponectin 1363:1381 control adiponectin 1363:1381 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 5 15 theme acid 797:800 arg1 removal 779:785 Enzymatic removal 769:785 Enzymatic removal of sialic acid or its underlying O-linked sugars 769:834 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 8 16 theme desialylated 1300:1311 arg1 adiponectin 1313:1323 desialylated adiponectin 1300:1323 desialylated adiponectin 1300:1323 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 4 17 theme variable 731:738 arg1 domain 740:745 the variable domain 727:745 the variable domain in human adiponectin 727:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 7 18 gly desialylated 1146:1157 arg1 adiponectin 1159:1169 desialylated adiponectin 1146:1169 desialylated adiponectin 1146:1169 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 7 19 theme desialylated 1146:1157 arg1 adiponectin 1159:1169 desialylated adiponectin 1146:1169 desialylated adiponectin 1146:1169 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 9 20 theme asialoglycoprotein 1609:1626 arg1 receptor 1628:1635 the hepatic asialoglycoprotein receptor 1597:1635 the hepatic asialoglycoprotein receptor 1597:1635 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 1 21 theme adipocyte-secreted 148:165 arg1 Adiponectin 130:140 Adiponectin 130:140 Adiponectin 130:140 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 1 21 theme adipocyte-secreted 148:165 arg1 hormone 188:194 an adipocyte-secreted, insulin-sensitizing hormone 145:194 an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes 145:287 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 2 22 from importance 325:334 arg1 secretion 474:482 secretion 474:482 secretion 474:482 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 2 22 from importance 325:334 arg1 function 489:496 function 489:496 function 489:496 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 2 22 from importance 325:334 arg1 oligomerization 457:471 adiponectin oligomerization 445:471 adiponectin oligomerization 445:471 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 11 23 theme important 1925:1933 arg1 role 1935:1938 an important role 1922:1938 an important role for sialic acid content in the regulation of circulating adiponectin levels 1922:2014 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 5 24 theme multimer 863:870 arg1 composition 872:882 adiponectin multimer composition 851:882 adiponectin multimer composition 851:882 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 5 25 theme O-linked 820:827 arg1 sugars 829:834 its underlying O-linked sugars 805:834 its underlying O-linked sugars 805:834 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 8 26 theme circulating 1457:1467 arg1 adiponectin 1469:1479 circulating adiponectin 1457:1479 circulating adiponectin 1457:1479 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 2 27 theme adiponectin 445:455 arg1 oligomerization 457:471 adiponectin oligomerization 445:471 adiponectin oligomerization 445:471 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 4 28 dep demonstrated 627:638 arg1 Using 582:586 Using 582:586 Using a variety of biochemical approaches 582:622 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 0 29 theme Sialic 0:5 arg1 modification 12:23 Sialic acid modification 0:23 Sialic acid modification of adiponectin 0:38 Sialic acid modification of adiponectin is not required for multimerization or secretion but determines half-life in circulation. 19855092 7 30 from adiponectin 1197:1207 arg1 mice 1261:1264 adiponectin(-/-) mice 1244:1264 adiponectin(-/-) mice 1244:1264 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 7 30 from adiponectin 1197:1207 arg1 myotubes 1215:1222 L6 myotubes 1212:1222 L6 myotubes 1212:1222 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 7 30 from adiponectin 1197:1207 arg1 assays 1234:1239 acute assays 1228:1239 acute assays 1228:1239 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 10 31 theme control 1882:1888 arg1 adiponectin 1890:1900 control adiponectin 1882:1900 control adiponectin 1882:1900 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 7 32 theme -/- 1256:1258 arg1 mice 1261:1264 adiponectin(-/-) mice 1244:1264 adiponectin(-/-) mice 1244:1264 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 0 33 theme adiponectin 28:38 arg1 modification 12:23 Sialic acid modification 0:23 Sialic acid modification of adiponectin 0:38 Sialic acid modification of adiponectin is not required for multimerization or secretion but determines half-life in circulation. 19855092 6 34 from Expression 885:894 arg1 cells 994:998 cells 994:998 cells defective in sialylation 994:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 10 35 theme chronic 1653:1659 arg1 administration 1661:1674 chronic administration 1653:1674 chronic administration in adiponectin(-/-) mice 1653:1699 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 10 36 theme tolerance 1852:1860 arg1 tests 1862:1866 glucose and insulin tolerance tests 1832:1866 glucose and insulin tolerance tests observed with control adiponectin 1832:1900 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 7 37 theme adiponectin 1244:1254 arg1 mice 1261:1264 adiponectin(-/-) mice 1244:1264 adiponectin(-/-) mice 1244:1264 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 6 38 theme modified 940:947 arg1 residues 953:960 the modified Thr residues 936:960 the modified Thr residues 936:960 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 7 39 theme L6 1212:1213 arg1 myotubes 1215:1222 L6 myotubes 1212:1222 L6 myotubes 1212:1222 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 10 40 theme -/- 1691:1693 arg1 mice 1696:1699 adiponectin(-/-) mice 1679:1699 adiponectin(-/-) mice 1679:1699 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 4 41 theme biochemical 601:611 arg1 approaches 613:622 biochemical approaches 601:622 biochemical approaches 601:622 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 3 42 theme functional 526:535 arg1 characterization 537:552 the first functional characterization 516:552 the first functional characterization of adiponectin sialylation 516:579 Here we describe the first functional characterization of adiponectin sialylation. 19855092 9 43 theme desialylated 1492:1503 arg1 adiponectin 1505:1515 desialylated adiponectin 1492:1515 desialylated adiponectin 1492:1515 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 5 44 link O-linked 820:827 arg1 sugars 829:834 its underlying O-linked sugars 805:834 its underlying O-linked sugars 805:834 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 4 45 theme O-linked 691:698 arg1 glycans 700:706 previously unidentified O-linked glycans 667:706 previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin 667:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 6 46 from defective 1000:1008 arg1 sialylation 1013:1023 sialylation 1013:1023 sialylation 1013:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 2 47 theme lysine 406:411 arg1 hydroxylation/glycosylation 413:439 lysine hydroxylation/glycosylation 406:439 lysine hydroxylation/glycosylation 406:439 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 6 48 from sialylation 1013:1023 arg1 defective 1000:1008 defective 1000:1008 defective 1000:1008 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 5 49 theme adiponectin 851:861 arg1 composition 872:882 adiponectin multimer composition 851:882 adiponectin multimer composition 851:882 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 7 50 theme acute 1228:1232 arg1 assays 1234:1239 acute assays 1228:1239 acute assays 1228:1239 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 3 51 theme sialylation 569:579 arg1 characterization 537:552 the first functional characterization 516:552 the first functional characterization of adiponectin sialylation 516:579 Here we describe the first functional characterization of adiponectin sialylation. 19855092 1 52 theme resistance 265:274 arg1 conditions 243:252 conditions 243:252 conditions of insulin resistance and diabetes 243:287 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 6 53 theme defective 1000:1008 arg1 cells 994:998 cells 994:998 cells defective in sialylation 994:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 4 54 link O-linked 691:698 arg1 glycans 700:706 previously unidentified O-linked glycans 667:706 previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin 667:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 10 55 theme steady-state 1701:1712 arg1 levels 1714:1719 steady-state levels 1701:1719 steady-state levels of desialylated adiponectin 1701:1747 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 1 56 theme diabetes 280:287 arg1 conditions 243:252 conditions 243:252 conditions of insulin resistance and diabetes 243:287 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 11 57 theme adiponectin 1997:2007 arg1 levels 2009:2014 circulating adiponectin levels 1985:2014 circulating adiponectin levels 1985:2014 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 11 58 theme levels 2009:2014 arg1 regulation 1971:1980 the regulation 1967:1980 the regulation of circulating adiponectin levels 1967:2014 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 6 59 theme mutant 899:904 arg1 forms 906:910 mutant forms 899:910 mutant forms of adiponectin (lacking the modified Thr residues) 899:961 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 1 60 theme hormone 188:194 arg1 levels 212:217 the circulating levels 196:217 the circulating levels 196:217 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 6 61 theme wild-type 969:977 arg1 adiponectin 979:989 wild-type adiponectin 969:989 wild-type adiponectin in cells defective in sialylation 969:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 4 62 theme domain 740:745 arg1 residues 715:722 Thr residues 711:722 Thr residues of the variable domain in human adiponectin 711:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 4 62 theme domain 740:745 arg1 domain 740:745 the variable domain 727:745 the variable domain in human adiponectin 727:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 10 63 theme adiponectin 1737:1747 arg1 levels 1714:1719 steady-state levels 1701:1719 steady-state levels of desialylated adiponectin 1701:1747 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 8 64 gly desialylated 1300:1311 arg1 adiponectin 1313:1323 desialylated adiponectin 1300:1323 desialylated adiponectin 1300:1323 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 2 65 theme posttranslational 339:355 arg1 hydroxylation 388:400 proline hydroxylation 380:400 proline hydroxylation 380:400 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 2 65 theme posttranslational 339:355 arg1 modifications 357:369 posttranslational modifications 339:369 posttranslational modifications 339:369 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 2 65 theme posttranslational 339:355 arg1 hydroxylation/glycosylation 413:439 lysine hydroxylation/glycosylation 406:439 lysine hydroxylation/glycosylation 406:439 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 4 66 from domain 740:745 arg1 adiponectin 756:766 human adiponectin 750:766 human adiponectin 750:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 1 67 theme circulating 200:210 arg1 levels 212:217 the circulating levels 196:217 the circulating levels 196:217 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 4 68 theme human 750:754 arg1 adiponectin 756:766 human adiponectin 750:766 human adiponectin 750:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 5 69 theme sialic 790:795 arg1 acid 797:800 sialic acid 790:800 sialic acid 790:800 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 8 70 theme plasma 1280:1285 arg1 clearance 1287:1295 plasma clearance 1280:1295 plasma clearance of desialylated adiponectin 1280:1323 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 10 71 theme adiponectin 1679:1689 arg1 mice 1696:1699 adiponectin(-/-) mice 1679:1699 adiponectin(-/-) mice 1679:1699 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 10 72 theme control 1765:1771 arg1 adiponectin 1773:1783 control adiponectin 1765:1783 control adiponectin 1765:1783 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 10 73 theme insulin 1844:1850 arg1 tests 1862:1866 glucose and insulin tolerance tests 1832:1866 glucose and insulin tolerance tests observed with control adiponectin 1832:1900 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 7 74 theme adiponectin 1159:1169 arg1 Activity 1134:1141 Activity 1134:1141 Activity of desialylated adiponectin 1134:1169 Activity of desialylated adiponectin was comparable to control adiponectin in L6 myotubes and acute assays in adiponectin(-/-) mice. 19855092 4 75 from residues 715:722 arg1 adiponectin 756:766 human adiponectin 750:766 human adiponectin 750:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 6 76 theme multimer 1044:1051 arg1 formation 1053:1061 multimer formation 1044:1061 multimer formation 1044:1061 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 11 77 from role 1935:1938 arg1 regulation 1971:1980 the regulation 1967:1980 the regulation of circulating adiponectin levels 1967:2014 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 9 78 theme hepatic 1601:1607 arg1 receptor 1628:1635 the hepatic asialoglycoprotein receptor 1597:1635 the hepatic asialoglycoprotein receptor 1597:1635 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 8 79 theme adiponectin 1313:1323 arg1 clearance 1287:1295 plasma clearance 1280:1295 plasma clearance of desialylated adiponectin 1280:1323 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 4 80 gly sialylation 645:655 arg2 glycans 700:706 previously unidentified O-linked glycans 667:706 previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin 667:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 11 81 theme circulating 1985:1995 arg1 levels 2009:2014 circulating adiponectin levels 1985:2014 circulating adiponectin levels 1985:2014 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 9 82 gly asialoglycoprotein 1609:1626 arg1 asialoglycoprotein 1609:1626 the hepatic asialoglycoprotein receptor 1597:1635 the hepatic asialoglycoprotein receptor 1597:1635 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 4 83 from glycans 700:706 arg1 residues 715:722 Thr residues 711:722 Thr residues of the variable domain in human adiponectin 711:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 4 83 from glycans 700:706 arg1 domain 740:745 the variable domain 727:745 the variable domain in human adiponectin 727:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 6 84 theme structural 1095:1104 arg1 role 1106:1109 a structural role 1093:1109 a structural role for this modification 1093:1131 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 5 85 theme underlying 809:818 arg1 sugars 829:834 its underlying O-linked sugars 805:834 its underlying O-linked sugars 805:834 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 10 86 theme glucose 1832:1838 arg1 tests 1862:1866 glucose and insulin tolerance tests 1832:1866 glucose and insulin tolerance tests observed with control adiponectin 1832:1900 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 5 87 theme sugars 829:834 arg1 removal 779:785 Enzymatic removal 769:785 Enzymatic removal of sialic acid or its underlying O-linked sugars 769:834 Enzymatic removal of sialic acid or its underlying O-linked sugars did not affect adiponectin multimer composition. 19855092 10 88 gly desialylated 1724:1735 arg1 adiponectin 1737:1747 desialylated adiponectin 1724:1747 desialylated adiponectin 1724:1747 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 11 89 theme adiponectin 2084:2094 arg1 sialylation/desialylation 2096:2120 adiponectin sialylation/desialylation 2084:2120 adiponectin sialylation/desialylation 2084:2120 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 0 90 theme acid 7:10 arg1 modification 12:23 Sialic acid modification 0:23 Sialic acid modification of adiponectin 0:38 Sialic acid modification of adiponectin is not required for multimerization or secretion but determines half-life in circulation. 19855092 11 91 theme sialic 1944:1949 arg1 content 1956:1962 sialic acid content 1944:1962 sialic acid content 1944:1962 These data suggest an important role for sialic acid content in the regulation of circulating adiponectin levels and highlight the importance of understanding mechanisms regulating adiponectin sialylation/desialylation. 19855092 8 92 theme adiponectin 1469:1479 arg1 half-life 1444:1452 the half-life 1440:1452 the half-life of circulating adiponectin 1440:1479 In contrast, plasma clearance of desialylated adiponectin was accelerated compared with that of control adiponectin, implicating a role for this modification in determining the half-life of circulating adiponectin. 19855092 1 93 dep adipocyte-secreted 148:165 arg1 insulin-sensitizing 168:186 insulin-sensitizing 168:186 insulin-sensitizing 168:186 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 3 94 theme first 520:524 arg1 characterization 537:552 the first functional characterization 516:552 the first functional characterization of adiponectin sialylation 516:579 Here we describe the first functional characterization of adiponectin sialylation. 19855092 10 95 from administration 1661:1674 arg1 mice 1696:1699 adiponectin(-/-) mice 1679:1699 adiponectin(-/-) mice 1679:1699 Finally, after chronic administration in adiponectin(-/-) mice steady-state levels of desialylated adiponectin were lower than control adiponectin and failed to recapitulate the improvements in glucose and insulin tolerance tests observed with control adiponectin. 19855092 0 96 mod modification 12:23 arg3 acid 7:10 Sialic acid modification 0:23 Sialic acid modification of adiponectin 0:38 Sialic acid modification of adiponectin is not required for multimerization or secretion but determines half-life in circulation. 19855092 0 96 mod modification 12:23 arg1 adiponectin 28:38 adiponectin 28:38 adiponectin 28:38 Sialic acid modification of adiponectin is not required for multimerization or secretion but determines half-life in circulation. 19855092 9 97 gly desialylated 1492:1503 arg1 adiponectin 1505:1515 desialylated adiponectin 1492:1515 desialylated adiponectin 1492:1515 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 6 98 theme Thr 949:951 arg1 residues 953:960 the modified Thr residues 936:960 the modified Thr residues 936:960 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 4 99 theme approaches 613:622 arg1 variety 590:596 a variety 588:596 a variety of biochemical approaches 588:622 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 4 99 theme approaches 613:622 arg1 approaches 613:622 biochemical approaches 601:622 biochemical approaches 601:622 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 3 100 theme adiponectin 557:567 arg1 sialylation 569:579 adiponectin sialylation 557:579 adiponectin sialylation 557:579 Here we describe the first functional characterization of adiponectin sialylation. 19855092 2 101 theme modifications 357:369 arg1 importance 325:334 the importance 321:334 the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function 321:496 Previous work has demonstrated the importance of posttranslational modifications, such as proline hydroxylation and lysine hydroxylation/glycosylation, in adiponectin oligomerization, secretion, and function. 19855092 9 102 theme adiponectin 1505:1515 arg1 Uptake 1482:1487 Uptake 1482:1487 Uptake of desialylated adiponectin by isolated primary rat hepatocytes 1482:1551 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 19855092 4 103 theme Thr 711:713 arg1 residues 715:722 Thr residues 711:722 Thr residues of the variable domain in human adiponectin 711:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 4 103 theme Thr 711:713 arg1 domain 740:745 the variable domain 727:745 the variable domain in human adiponectin 727:766 Using a variety of biochemical approaches we demonstrated that sialylation occurs on previously unidentified O-linked glycans on Thr residues of the variable domain in human adiponectin. 19855092 6 104 from cells 994:998 arg1 Expression 885:894 Expression 885:894 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation 885:1023 Expression of mutant forms of adiponectin (lacking the modified Thr residues) or of wild-type adiponectin in cells defective in sialylation did not compromise multimer formation or secretion, arguing against a structural role for this modification. 19855092 1 105 theme insulin 257:263 arg1 resistance 265:274 insulin resistance 257:274 insulin resistance 257:274 Adiponectin is an adipocyte-secreted, insulin-sensitizing hormone the circulating levels of which are reduced in conditions of insulin resistance and diabetes. 19855092 9 106 theme isolated 1520:1527 arg1 hepatocytes 1541:1551 isolated primary rat hepatocytes 1520:1551 isolated primary rat hepatocytes 1520:1551 Uptake of desialylated adiponectin by isolated primary rat hepatocytes was also accelerated, suggesting a role for the hepatic asialoglycoprotein receptor. 6510521 5 0 theme acid 739:742 arg1 .. 762:763 acid hydrolysis--bonds .. 739:763 acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 739:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 3 1 dep fragments 544:552 arg1 CB3 562:564 CB3 562:564 CB3 562:564 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 3 1 dep fragments 544:552 arg1 CB2 554:556 CB2 554:556 CB2 554:556 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 3 1 dep fragments 544:552 arg1 fragments 544:552 fragments CB2 and CB3 544:564 fragments CB2 and CB3 544:564 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 4 2 dep attached 640:647 arg1 positions 650:658 positions 1,41,164,217 and probably 223 650:688 positions 1,41,164,217 and probably 223 650:688 Fragments CB1-CB4 include 5 sites, where hexosamine oligosaccharides are attached (positions 1,41,164,217 and probably 223). 6510521 3 3 dep residues 500:507 arg1 165/170 521:527 165/170 521:527 165/170 521:527 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 3 3 dep residues 500:507 arg1 126/130 509:515 126/130 509:515 126/130 509:515 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 5 4 from .. 762:763 arg1 positions 782:790 positions 20/21 and 187/188 782:808 positions 20/21 and 187/188 782:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 5 5 theme sequenced 699:707 arg1 region 709:714 the sequenced region 695:714 the sequenced region 695:714 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 1 6 from residues 125:132 arg1 digestion 82:90 Cyanogen bromide digestion 65:90 Cyanogen bromide digestion of hemopexin at its 6 methionine residues 65:132 Cyanogen bromide digestion of hemopexin at its 6 methionine residues results in 7 fragments (CB1-CB7) partially connected by disulfide bridges. 6510521 3 7 theme intramolecular 444:457 arg1 bonds 469:473 intramolecular disulfide bonds 444:473 intramolecular disulfide bonds 444:473 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 3 8 theme disulfide 459:467 arg1 bonds 469:473 intramolecular disulfide bonds 444:473 intramolecular disulfide bonds 444:473 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 2 9 theme hemopexin 365:373 arg1 region 349:354 the N-terminal region 334:354 the N-terminal region of human hemopexin 334:373 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 5 10 theme Asp--Pro 766:773 arg1 .. 775:776 Asp--Pro .. 766:776 acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 739:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 5 11 theme hydrolysis--bonds 744:760 arg1 .. 762:763 acid hydrolysis--bonds .. 739:763 acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 739:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 1 12 theme Cyanogen 65:72 arg1 bromide 74:80 bromide 74:80 bromide 74:80 Cyanogen bromide digestion of hemopexin at its 6 methionine residues results in 7 fragments (CB1-CB7) partially connected by disulfide bridges. 6510521 2 13 theme human 359:363 arg1 hemopexin 365:373 human hemopexin 359:373 human hemopexin 359:373 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 0 14 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the N-terminal region of human hemopexin 0:62 Amino acid sequence of the N-terminal region of human hemopexin. 6510521 1 15 theme bromide 74:80 arg1 digestion 82:90 Cyanogen bromide digestion 65:90 Cyanogen bromide digestion of hemopexin at its 6 methionine residues 65:132 Cyanogen bromide digestion of hemopexin at its 6 methionine residues results in 7 fragments (CB1-CB7) partially connected by disulfide bridges. 6510521 0 16 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the N-terminal region of human hemopexin 0:62 Amino acid sequence of the N-terminal region of human hemopexin. 6510521 5 17 dep .. 762:763 arg1 .. 775:776 Asp--Pro .. 766:776 acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 739:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 2 18 theme region 349:354 arg1 sequence 322:329 a continuous amino acid sequence 298:329 a continuous amino acid sequence 298:329 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 2 19 theme enzyme 281:286 arg1 cleavage 288:295 their enzyme cleavage 275:295 their enzyme cleavage 275:295 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 2 20 theme amino 391:395 arg1 residues 402:409 220 amino acid residues 387:409 220 amino acid residues 387:409 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 2 21 theme acid 397:400 arg1 residues 402:409 220 amino acid residues 387:409 220 amino acid residues 387:409 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 6 22 theme sequence 895:902 arg1 heterogeneity 904:916 no sequence heterogeneity 892:916 no sequence heterogeneity 892:916 In spite of the fact that pooled material of many donors was studied, no sequence heterogeneity was discovered. 6510521 1 23 theme hemopexin 95:103 arg1 digestion 82:90 Cyanogen bromide digestion 65:90 Cyanogen bromide digestion of hemopexin at its 6 methionine residues 65:132 Cyanogen bromide digestion of hemopexin at its 6 methionine residues results in 7 fragments (CB1-CB7) partially connected by disulfide bridges. 6510521 3 24 theme half-cystine 487:498 arg1 residues 500:507 half-cystine residues 126/130 and 165/170 487:527 half-cystine residues 126/130 and 165/170 487:527 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 0 25 theme N-terminal 27:36 arg1 region 38:43 the N-terminal region 23:43 the N-terminal region of human hemopexin 23:62 Amino acid sequence of the N-terminal region of human hemopexin. 6510521 3 26 theme bonds 469:473 arg1 presence 432:439 The presence 428:439 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, 428:528 The presence of intramolecular disulfide bonds, connecting half-cystine residues 126/130 and 165/170, was proved in fragments CB2 and CB3. 6510521 5 27 located found 815:819 arg1 region 709:714 the sequenced region 695:714 the sequenced region 695:714 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 5 27 located found 815:819 arg2 sites 720:724 two sites 716:724 two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 716:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 2 28 theme sequence 212:219 arg1 studies 221:227 sequence studies 212:227 sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage 212:295 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 2 29 dep fragments 232:240 arg1 peptides 254:261 peptides 254:261 peptides 254:261 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 2 29 dep fragments 232:240 arg1 CB1-CB4 242:248 CB1-CB4 242:248 CB1-CB4 242:248 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 4 30 theme hexosamine 608:617 arg1 oligosaccharides 619:634 hexosamine oligosaccharides 608:634 hexosamine oligosaccharides 608:634 Fragments CB1-CB4 include 5 sites, where hexosamine oligosaccharides are attached (positions 1,41,164,217 and probably 223). 6510521 1 31 theme disulfide 190:198 arg1 bridges 200:206 disulfide bridges 190:206 disulfide bridges 190:206 Cyanogen bromide digestion of hemopexin at its 6 methionine residues results in 7 fragments (CB1-CB7) partially connected by disulfide bridges. 6510521 5 32 theme sensitive 726:734 arg1 sites 720:724 two sites 716:724 two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 716:808 In the sequenced region two sites sensitive to acid hydrolysis--bonds ... Asp--Pro ... in positions 20/21 and 187/188 were found. 6510521 2 33 theme acid 317:320 arg1 sequence 322:329 a continuous amino acid sequence 298:329 a continuous amino acid sequence 298:329 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 0 34 theme region 38:43 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the N-terminal region of human hemopexin 0:62 Amino acid sequence of the N-terminal region of human hemopexin. 6510521 2 35 theme amino 311:315 arg1 sequence 322:329 a continuous amino acid sequence 298:329 a continuous amino acid sequence 298:329 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 6 36 theme donors 872:877 arg1 material 855:862 pooled material 848:862 pooled material of many donors 848:877 In spite of the fact that pooled material of many donors was studied, no sequence heterogeneity was discovered. 6510521 2 37 theme continuous 300:309 arg1 sequence 322:329 a continuous amino acid sequence 298:329 a continuous amino acid sequence 298:329 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 4 38 theme CB1-CB4 577:583 arg1 Fragments 567:575 Fragments 567:575 Fragments CB1-CB4 567:583 Fragments CB1-CB4 include 5 sites, where hexosamine oligosaccharides are attached (positions 1,41,164,217 and probably 223). 6510521 4 39 dep positions 650:658 arg1 1,41,164,217 660:671 1,41,164,217 660:671 1,41,164,217 660:671 Fragments CB1-CB4 include 5 sites, where hexosamine oligosaccharides are attached (positions 1,41,164,217 and probably 223). 6510521 4 39 dep positions 650:658 arg1 223 686:688 223 686:688 223 686:688 Fragments CB1-CB4 include 5 sites, where hexosamine oligosaccharides are attached (positions 1,41,164,217 and probably 223). 6510521 2 40 theme fragments 232:240 arg1 studies 221:227 sequence studies 212:227 sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage 212:295 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 1 41 theme methionine 114:123 arg1 residues 125:132 6 methionine residues 112:132 its 6 methionine residues 108:132 Cyanogen bromide digestion of hemopexin at its 6 methionine residues results in 7 fragments (CB1-CB7) partially connected by disulfide bridges. 6510521 0 42 theme hemopexin 54:62 arg1 region 38:43 the N-terminal region 23:43 the N-terminal region of human hemopexin 23:62 Amino acid sequence of the N-terminal region of human hemopexin. 6510521 2 43 theme N-terminal 338:347 arg1 region 349:354 the N-terminal region 334:354 the N-terminal region of human hemopexin 334:373 By sequence studies of fragments CB1-CB4 and peptides prepared by their enzyme cleavage, a continuous amino acid sequence of the N-terminal region of human hemopexin, comprising 220 amino acid residues, was determined. 6510521 6 44 theme pooled 848:853 arg1 material 855:862 pooled material 848:862 pooled material of many donors 848:877 In spite of the fact that pooled material of many donors was studied, no sequence heterogeneity was discovered. 6510521 0 45 theme human 48:52 arg1 hemopexin 54:62 human hemopexin 48:62 human hemopexin 48:62 Amino acid sequence of the N-terminal region of human hemopexin. 6510521 6 46 theme many 867:870 arg1 donors 872:877 many donors 867:877 many donors 867:877 In spite of the fact that pooled material of many donors was studied, no sequence heterogeneity was discovered. 23803857 6 0 dep = 1123:1123 arg1 M 1139:1139 6.7 × 10(-10) M 1125:1139 6.7 × 10(-10) M 1125:1139 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 4 1 theme lectin-like 775:785 arg1 dimers 787:792 other C-type lectin-like dimers 762:792 other C-type lectin-like dimers 762:792 Whereas KACL forms a homodimer resembling other C-type lectin-like dimers, NKp65 is monomeric. 23803857 0 2 link linked 82:87 arg1 recognition 89:99 genetically linked recognition 70:99 genetically linked recognition in natural killer gene complex 70:130 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 1 3 theme killer 145:150 arg1 NKC 171:173 NKC 171:173 NKC 171:173 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 3 theme killer 145:150 arg1 complex 162:168 The natural killer (NK) gene complex 133:168 The natural killer (NK) gene complex (NKC) 133:174 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 3 theme killer 145:150 arg1 receptors 212:220 numerous C-type lectin-like receptors 184:220 numerous C-type lectin-like receptors that govern the activity of NK cells 184:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 7 4 theme other 1419:1423 arg1 pairs 1444:1448 other genetically linked pairs 1419:1448 other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL 1419:1509 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 6 5 theme cell 1209:1212 arg1 activation 1214:1223 cell activation 1209:1223 cell activation 1209:1223 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 3 6 theme activating 576:585 arg1 NKp65 566:570 human NKp65 560:570 human NKp65 560:570 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 3 6 theme activating 576:585 arg1 receptor 587:594 an activating receptor 573:594 an activating receptor implicated in the immunosurveillance of skin 573:639 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 3 7 theme human 560:564 arg1 NKp65 566:570 human NKp65 560:570 human NKp65 560:570 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 3 7 theme human 560:564 arg1 receptor 587:594 an activating receptor 573:594 an activating receptor implicated in the immunosurveillance of skin 573:639 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 7 8 theme receptor-ligand 1281:1295 arg1 complex 1297:1303 an NKC-encoded receptor-ligand complex 1266:1303 an NKC-encoded receptor-ligand complex 1266:1303 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 0 9 theme linked 82:87 arg1 recognition 89:99 genetically linked recognition 70:99 genetically linked recognition in natural killer gene complex 70:130 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 7 10 theme complex 1297:1303 arg1 structure 1253:1261 This previously unreported structure 1226:1261 This previously unreported structure 1226:1261 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 6 11 theme interaction 1059:1069 arg1 affinity 1032:1039 the exceptionally high affinity 1009:1039 the exceptionally high affinity of the NKp65-KACL interaction 1009:1069 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 0 12 theme natural 104:110 arg1 killer 112:117 natural killer 104:117 natural killer gene complex 104:130 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 3 13 theme skin 636:639 arg1 immunosurveillance 614:631 the immunosurveillance 610:631 the immunosurveillance of skin 610:639 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 5 14 theme NKp65-KACL 839:848 arg1 complex 850:856 the NKp65-KACL complex 835:856 the NKp65-KACL complex 835:856 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 6 15 theme NKp65-KACL 1048:1057 arg1 interaction 1059:1069 the NKp65-KACL interaction 1044:1069 the NKp65-KACL interaction 1044:1069 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 1 16 theme gene 157:160 arg1 NKC 171:173 NKC 171:173 NKC 171:173 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 16 theme gene 157:160 arg1 complex 162:168 The natural killer (NK) gene complex 133:168 The natural killer (NK) gene complex (NKC) 133:174 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 16 theme gene 157:160 arg1 receptors 212:220 numerous C-type lectin-like receptors 184:220 numerous C-type lectin-like receptors that govern the activity of NK cells 184:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 2 17 theme MHC-like 337:344 arg1 molecules 346:354 MHC or MHC-like molecules 330:354 molecules 346:354 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 5 18 from mode 827:830 arg1 complex 850:856 the NKp65-KACL complex 835:856 the NKp65-KACL complex 835:856 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 5 19 theme monomeric 870:878 arg1 receptor 880:887 a monomeric receptor 868:887 a monomeric receptor 868:887 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 6 20 theme NKp65 1192:1196 arg1 nature 1182:1187 the monomeric nature 1168:1187 the monomeric nature of NKp65 1168:1196 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 4 21 theme other 762:766 arg1 dimers 787:792 other C-type lectin-like dimers 762:792 other C-type lectin-like dimers 762:792 Whereas KACL forms a homodimer resembling other C-type lectin-like dimers, NKp65 is monomeric. 23803857 5 22 from those 943:947 arg1 distinct 929:936 distinct 929:936 distinct 929:936 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 5 22 from those 943:947 arg1 mode 827:830 The binding mode 815:830 The binding mode 815:830 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 3 23 dep ligand 667:672 arg1 KACL 713:716 KACL 713:716 KACL 713:716 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 3 23 dep ligand 667:672 arg1 lectin 705:710 keratinocyte-associated C-type lectin 674:710 its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL) 651:717 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 2 24 theme MHC 330:332 arg1 molecules 346:354 MHC or MHC-like molecules 330:354 molecules 346:354 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 7 25 theme mutational 1319:1328 arg1 analysis 1330:1337 mutational analysis 1319:1337 mutational analysis of the interface 1319:1354 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 7 26 theme interface 1346:1354 arg1 analysis 1330:1337 mutational analysis 1319:1337 mutational analysis of the interface 1319:1354 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 2 27 dep receptors 283:291 arg1 Ly49s 294:298 Ly49s 294:298 Ly49s 294:298 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 2 27 dep receptors 283:291 arg1 CD94/NKG2A 308:317 CD94/NKG2A 308:317 CD94/NKG2A 308:317 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 2 27 dep receptors 283:291 arg1 NKG2D 301:305 NKG2D 301:305 NKG2D 301:305 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 2 28 attach linked 464:469 arg2 they 443:446 they 443:446 they 443:446 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 2 28 attach linked 464:469 arg1 ligands 426:432 C-type lectin-like ligands 407:432 C-type lectin-like ligands to which they are genetically linked in the NKC 407:480 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 2 28 attach linked 464:469 arg3 NKC 478:480 the NKC 474:480 the NKC 474:480 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 4 29 theme C-type 768:773 arg1 dimers 787:792 other C-type lectin-like dimers 762:792 other C-type lectin-like dimers 762:792 Whereas KACL forms a homodimer resembling other C-type lectin-like dimers, NKp65 is monomeric. 23803857 1 30 theme NK 250:251 arg1 cells 253:257 NK cells 250:257 NK cells 250:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 0 31 theme gene 119:122 arg1 complex 124:130 natural killer gene complex 104:130 natural killer gene complex 104:130 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 6 32 theme high 1027:1030 arg1 affinity 1032:1039 the exceptionally high affinity 1009:1039 the exceptionally high affinity of the NKp65-KACL interaction 1009:1069 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 0 33 theme NKp65 13:17 arg1 Structure 0:8 Structure 0:8 Structure of NKp65 bound to its keratinocyte ligand 0:50 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 1 34 theme cells 253:257 arg1 activity 238:245 the activity 234:245 the activity of NK cells 234:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 0 35 theme killer 112:117 arg1 complex 124:130 natural killer gene complex 104:130 natural killer gene complex 104:130 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 7 36 theme unreported 1242:1251 arg1 structure 1253:1261 This previously unreported structure 1226:1261 This previously unreported structure 1226:1261 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 6 37 theme monomeric 1172:1180 arg1 nature 1182:1187 the monomeric nature 1168:1187 the monomeric nature of NKp65 1168:1196 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 6 38 theme = 1123:1123 arg1 KD 1120:1121 KD 1120:1121 KD = 6.7 × 10(-10) M 1120:1139 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 7 39 theme docking 1371:1377 arg1 applicable 1405:1414 applicable 1405:1414 applicable 1405:1414 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 7 39 theme docking 1371:1377 arg1 template 1379:1386 a docking template 1369:1386 a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL 1369:1509 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 3 40 theme keratinocyte-associated 674:696 arg1 KACL 713:716 KACL 713:716 KACL 713:716 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 3 40 theme keratinocyte-associated 674:696 arg1 lectin 705:710 keratinocyte-associated C-type lectin 674:710 its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL) 651:717 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 7 41 theme linked 1437:1442 arg1 pairs 1444:1448 other genetically linked pairs 1419:1448 other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL 1419:1509 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 3 42 theme C-type 698:703 arg1 KACL 713:716 KACL 713:716 KACL 713:716 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 3 42 theme C-type 698:703 arg1 lectin 705:710 keratinocyte-associated C-type lectin 674:710 its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL) 651:717 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 0 43 theme keratinocyte 32:43 arg1 ligand 45:50 its keratinocyte ligand 28:50 its keratinocyte ligand 28:50 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 3 44 theme NKC-encoded 655:665 arg1 ligand 667:672 its NKC-encoded ligand 651:672 its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL) 651:717 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 7 45 from pairs 1444:1448 arg1 NKRP1A-LLT1 1483:1493 NKRP1A-LLT1 1483:1493 NKRP1A-LLT1 1483:1493 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 7 45 from pairs 1444:1448 arg1 NKC 1457:1459 the NKC 1453:1459 the NKC 1453:1459 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 7 45 from pairs 1444:1448 arg1 Nkrp1-Clr 1472:1480 Nkrp1-Clr 1472:1480 Nkrp1-Clr 1472:1480 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 7 45 from pairs 1444:1448 arg1 NKp80-AICL 1500:1509 NKp80-AICL 1500:1509 NKp80-AICL 1500:1509 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 2 46 dep others 357:362 arg1 NKRP1A 372:377 NKRP1A 372:377 NKRP1A 372:377 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 5 47 theme binding 819:825 arg1 distinct 929:936 distinct 929:936 distinct 929:936 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 5 47 theme binding 819:825 arg1 mode 827:830 The binding mode 815:830 The binding mode 815:830 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 7 48 link linked 1437:1442 arg1 pairs 1444:1448 other genetically linked pairs 1419:1448 other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL 1419:1509 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 6 49 theme interaction 1101:1111 arg1 pairs 1113:1117 other cell-cell interaction pairs 1085:1117 other cell-cell interaction pairs (KD = 6.7 × 10(-10) M) 1085:1140 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 6 50 theme × 1129:1129 arg1 M 1139:1139 6.7 × 10(-10) M 1125:1139 6.7 × 10(-10) M 1125:1139 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 3 51 dep determined 532:541 arg1 bound 642:646 bound 642:646 determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL) 532:717 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 1 52 theme numerous 184:191 arg1 receptors 212:220 numerous C-type lectin-like receptors 184:220 numerous C-type lectin-like receptors that govern the activity of NK cells 184:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 52 theme numerous 184:191 arg1 complex 162:168 The natural killer (NK) gene complex 133:168 The natural killer (NK) gene complex (NKC) 133:174 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 7 53 theme NKC-encoded 1269:1279 arg1 complex 1297:1303 an NKC-encoded receptor-ligand complex 1266:1303 an NKC-encoded receptor-ligand complex 1266:1303 This previously unreported structure of an NKC-encoded receptor-ligand complex, coupled with mutational analysis of the interface, establishes a docking template that is directly applicable to other genetically linked pairs in the NKC, including Nkrp1-Clr, NKRP1A-LLT1, and NKp80-AICL. 23803857 0 54 from recognition 89:99 arg1 complex 124:130 natural killer gene complex 104:130 natural killer gene complex 104:130 Structure of NKp65 bound to its keratinocyte ligand reveals basis for genetically linked recognition in natural killer gene complex. 23803857 6 55 theme cell-cell 1091:1099 arg1 pairs 1113:1117 other cell-cell interaction pairs 1085:1117 other cell-cell interaction pairs (KD = 6.7 × 10(-10) M) 1085:1140 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 6 56 dep pairs 1113:1117 arg1 KD 1120:1121 KD 1120:1121 KD = 6.7 × 10(-10) M 1120:1139 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 1 57 theme C-type 193:198 arg1 receptors 212:220 numerous C-type lectin-like receptors 184:220 numerous C-type lectin-like receptors that govern the activity of NK cells 184:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 57 theme C-type 193:198 arg1 complex 162:168 The natural killer (NK) gene complex 133:168 The natural killer (NK) gene complex (NKC) 133:174 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 2 58 theme lectin-like 414:424 arg1 ligands 426:432 C-type lectin-like ligands 407:432 C-type lectin-like ligands to which they are genetically linked in the NKC 407:480 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 3 59 theme NKp65 566:570 arg1 structure 547:555 the structure 543:555 the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, 543:640 To understand the basis for this recognition, we determined the structure of human NKp65, an activating receptor implicated in the immunosurveillance of skin, bound to its NKC-encoded ligand keratinocyte-associated C-type lectin (KACL). 23803857 6 60 theme other 1085:1089 arg1 pairs 1113:1117 other cell-cell interaction pairs 1085:1117 other cell-cell interaction pairs (KD = 6.7 × 10(-10) M) 1085:1140 The structure explains the exceptionally high affinity of the NKp65-KACL interaction compared with other cell-cell interaction pairs (KD = 6.7 × 10(-10) M), which may compensate for the monomeric nature of NKp65 to achieve cell activation. 23803857 1 61 theme lectin-like 200:210 arg1 receptors 212:220 numerous C-type lectin-like receptors 184:220 numerous C-type lectin-like receptors that govern the activity of NK cells 184:257 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 61 theme lectin-like 200:210 arg1 complex 162:168 The natural killer (NK) gene complex 133:168 The natural killer (NK) gene complex (NKC) 133:174 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 2 62 theme C-type 407:412 arg1 ligands 426:432 C-type lectin-like ligands 407:432 C-type lectin-like ligands to which they are genetically linked in the NKC 407:480 Although some of these receptors (Ly49s, NKG2D, CD94/NKG2A) recognize MHC or MHC-like molecules, others (Nkrp1, NKRP1A, NKp80, NKp65) instead bind C-type lectin-like ligands to which they are genetically linked in the NKC. 23803857 5 63 theme dimeric 899:905 arg1 ligand 907:912 a dimeric ligand 897:912 a dimeric ligand 897:912 The binding mode in the NKp65-KACL complex, in which a monomeric receptor engages a dimeric ligand, is completely distinct from those used by Ly49s, NKG2D, or CD94/NKG2A. 23803857 1 64 theme natural 137:143 arg1 NK 153:154 NK 153:154 NK 153:154 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 23803857 1 64 theme natural 137:143 arg1 killer 145:150 The natural killer 133:150 The natural killer (NK) gene complex (NKC) 133:174 The natural killer (NK) gene complex (NKC) encodes numerous C-type lectin-like receptors that govern the activity of NK cells. 20802483 0 0 with assembly 29:36 arg1 receptors 71:79 its receptors 67:79 its receptors 67:79 Structural insights into the assembly and activation of IL-1β with its receptors. 20802483 5 1 theme Biochemical 544:554 arg1 analysis 556:563 Biochemical analysis 544:563 Biochemical analysis 544:563 Biochemical analysis demonstrated that IL-1β-IL-1RI and IL-1β-IL-1RII interacted similarly with IL-1RAcP. 20802483 1 2 theme inflammation 130:141 arg1 orchestrator 114:125 a key orchestrator 108:125 a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP) 108:267 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 2 theme inflammation 130:141 arg1 Interleukin 82:92 Interleukin 1β 82:95 Interleukin 1β (IL-1β) 82:103 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 7 3 theme IL-1 835:838 arg1 receptor 840:847 the IL-1 receptor 831:847 the IL-1 receptor 831:847 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 7 4 theme IL-1 917:920 arg1 family 922:927 the IL-1 family 913:927 the IL-1 family of cytokines 913:940 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 7 5 theme receptor 840:847 arg1 activation 817:826 activation 817:826 activation 817:826 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 7 5 theme receptor 840:847 arg1 assembly 804:811 assembly 804:811 assembly 804:811 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 6 6 theme loops 687:691 arg1 importance 669:678 the importance 665:678 the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism 665:758 It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. 20802483 1 7 theme defense 152:158 arg1 orchestrator 114:125 a key orchestrator 108:125 a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP) 108:267 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 7 theme defense 152:158 arg1 Interleukin 82:92 Interleukin 1β 82:95 Interleukin 1β (IL-1β) 82:103 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 3 8 theme crystal 389:395 arg1 structure 397:405 the crystal structure 385:405 the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP 385:468 Here we present the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. 20802483 1 9 theme host 147:150 arg1 defense 152:158 host defense 147:158 host defense 147:158 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 10 theme IL-1 226:229 arg1 IL-1RAcP 259:266 IL-1RAcP 259:266 IL-1RAcP 259:266 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 10 theme IL-1 226:229 arg1 protein 250:256 IL-1 receptor accessory protein 226:256 IL-1 receptor accessory protein (IL-1RAcP) 226:267 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 2 11 theme signaling-competent 324:342 arg1 complex 344:350 the signaling-competent complex 320:350 the signaling-competent complex 320:350 How IL-1RAcP is recruited by IL-1β-IL-1RI to form the signaling-competent complex remains elusive. 20802483 0 12 theme Structural 0:9 arg1 insights 11:18 Structural insights 0:18 Structural insights into the assembly and activation of IL-1β with its receptors. 0:80 Structural insights into the assembly and activation of IL-1β with its receptors. 20802483 4 13 theme composite 497:505 arg1 surface 515:521 a composite binding surface 495:521 a composite binding surface to recruit IL-1RAcP 495:541 IL-1β-IL-1RII generated a composite binding surface to recruit IL-1RAcP. 20802483 1 14 theme receptor 231:238 arg1 IL-1RAcP 259:266 IL-1RAcP 259:266 IL-1RAcP 259:266 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 14 theme receptor 231:238 arg1 protein 250:256 IL-1 receptor accessory protein 226:256 IL-1 receptor accessory protein (IL-1RAcP) 226:267 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 7 15 theme cytokines 932:940 arg1 family 922:927 the IL-1 family 913:927 the IL-1 family of cytokines 913:940 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 6 16 theme receptor 701:708 arg1 IL-1Ra 722:727 IL-1Ra 722:727 IL-1Ra 722:727 It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. 20802483 6 16 theme receptor 701:708 arg1 antagonist 710:719 IL-1 receptor antagonist 696:719 IL-1 receptor antagonist (IL-1Ra) 696:728 It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. 20802483 1 17 theme accessory 240:248 arg1 IL-1RAcP 259:266 IL-1RAcP 259:266 IL-1RAcP 259:266 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 17 theme accessory 240:248 arg1 protein 250:256 IL-1 receptor accessory protein 226:256 IL-1 receptor accessory protein (IL-1RAcP) 226:267 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 7 18 theme structural 783:792 arg1 basis 794:798 a structural basis 781:798 a structural basis for assembly and activation of the IL-1 receptor 781:847 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 4 19 theme binding 507:513 arg1 surface 515:521 a composite binding surface 495:521 a composite binding surface to recruit IL-1RAcP 495:541 IL-1β-IL-1RII generated a composite binding surface to recruit IL-1RAcP. 20802483 3 20 theme IL-1β 410:414 arg1 structure 397:405 the crystal structure 385:405 the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP 385:468 Here we present the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. 20802483 0 21 dep assembly 29:36 arg1 the 25:27 the 25:27 the 25:27 Structural insights into the assembly and activation of IL-1β with its receptors. 20802483 7 22 theme cytokine-receptor 869:885 arg1 architecture 887:898 a general cytokine-receptor architecture 859:898 a general cytokine-receptor architecture that governs the IL-1 family of cytokines 859:940 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 3 23 theme IL-1 425:428 arg1 type 439:442 IL-1 receptor type II 425:445 IL-1 receptor type II (IL-1RII) 425:455 Here we present the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. 20802483 3 23 theme IL-1 425:428 arg1 IL-1RII 448:454 IL-1RII 448:454 IL-1RII 448:454 Here we present the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. 20802483 1 24 theme IL-1 192:195 arg1 type 206:209 IL-1 receptor type I 192:211 IL-1 receptor type I (IL-1RI) 192:220 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 24 theme IL-1 192:195 arg1 IL-1RI 214:219 IL-1RI 214:219 IL-1RI 214:219 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 6 25 theme antagonist 710:719 arg1 loops 687:691 two loops 683:691 two loops of IL-1 receptor antagonist (IL-1Ra) 683:728 It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. 20802483 3 26 theme receptor 430:437 arg1 type 439:442 IL-1 receptor type II 425:445 IL-1 receptor type II (IL-1RII) 425:455 Here we present the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. 20802483 3 26 theme receptor 430:437 arg1 IL-1RII 448:454 IL-1RII 448:454 IL-1RII 448:454 Here we present the crystal structure of IL-1β bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. 20802483 1 27 theme receptor 197:204 arg1 type 206:209 IL-1 receptor type I 192:211 IL-1 receptor type I (IL-1RI) 192:220 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 27 theme receptor 197:204 arg1 IL-1RI 214:219 IL-1RI 214:219 IL-1RI 214:219 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 0 28 with activation 42:51 arg1 receptors 71:79 its receptors 67:79 its receptors 67:79 Structural insights into the assembly and activation of IL-1β with its receptors. 20802483 1 29 theme key 110:112 arg1 orchestrator 114:125 a key orchestrator 108:125 a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP) 108:267 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 1 29 theme key 110:112 arg1 Interleukin 82:92 Interleukin 1β 82:95 Interleukin 1β (IL-1β) 82:103 Interleukin 1β (IL-1β) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). 20802483 7 30 theme general 861:867 arg1 architecture 887:898 a general cytokine-receptor architecture 859:898 a general cytokine-receptor architecture that governs the IL-1 family of cytokines 859:940 Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines. 20802483 0 31 theme IL-1β 56:60 arg1 activation 42:51 activation 42:51 activation 42:51 Structural insights into the assembly and activation of IL-1β with its receptors. 20802483 0 31 theme IL-1β 56:60 arg1 assembly 29:36 assembly 29:36 assembly 29:36 Structural insights into the assembly and activation of IL-1β with its receptors. 20802483 6 32 theme IL-1 696:699 arg1 IL-1Ra 722:727 IL-1Ra 722:727 IL-1Ra 722:727 It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. 20802483 6 32 theme IL-1 696:699 arg1 antagonist 710:719 IL-1 receptor antagonist 696:719 IL-1 receptor antagonist (IL-1Ra) 696:728 It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. 34388369 0 0 theme signal 77:82 arg1 cleavage 92:99 signal peptide cleavage 77:99 signal peptide cleavage 77:99 Structure of the human signal peptidase complex reveals the determinants for signal peptide cleavage. 34388369 1 1 theme pre-proteins 278:289 arg1 variety 257:263 a large variety 249:263 a large variety of secretory pre-proteins with exquisite specificity 249:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 1 theme pre-proteins 278:289 arg1 pre-proteins 278:289 secretory pre-proteins 268:289 secretory pre-proteins with exquisite specificity 268:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 2 2 theme removal 440:446 arg1 details 410:416 the molecular details 396:416 the molecular details of SP recognition and removal 396:446 Although the determinants of this process have been established empirically, the molecular details of SP recognition and removal remain elusive. 34388369 4 3 theme structural 655:664 arg1 proteomics 666:675 electron cryo-microscopy and structural proteomics 626:675 electron cryo-microscopy and structural proteomics 626:675 We determined the atomic structures of both paralogs using electron cryo-microscopy and structural proteomics. 34388369 3 4 with paralogs 522:529 arg1 subunits 557:564 distinct proteolytic subunits 536:564 distinct proteolytic subunits 536:564 Here, we show that the human SPC exists in two functional paralogs with distinct proteolytic subunits. 34388369 1 5 theme exquisite 296:304 arg1 specificity 306:316 exquisite specificity 296:316 exquisite specificity 296:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 3 6 theme human 487:491 arg1 SPC 493:495 the human SPC 483:495 the human SPC 483:495 Here, we show that the human SPC exists in two functional paralogs with distinct proteolytic subunits. 34388369 1 7 with pre-proteins 278:289 arg1 specificity 306:316 exquisite specificity 296:316 exquisite specificity 296:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 8 theme signal 222:227 arg1 SPs 239:241 SPs 239:241 SPs 239:241 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 8 theme signal 222:227 arg1 peptides 229:236 signal peptides 222:236 signal peptides (SPs) 222:242 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 0 9 theme peptide 84:90 arg1 cleavage 92:99 signal peptide cleavage 77:99 signal peptide cleavage 77:99 Structure of the human signal peptidase complex reveals the determinants for signal peptide cleavage. 34388369 6 10 theme dynamics 854:861 arg1 simulations 863:873 Molecular dynamics simulations 844:873 Molecular dynamics simulations 844:873 Molecular dynamics simulations indicate that this unique architecture generates specificity for SPs based on the length of their hydrophobic segments. 34388369 6 11 theme Molecular 844:852 arg1 simulations 863:873 Molecular dynamics simulations 844:873 Molecular dynamics simulations 844:873 Molecular dynamics simulations indicate that this unique architecture generates specificity for SPs based on the length of their hydrophobic segments. 34388369 1 12 theme essential 143:151 arg1 complex 162:168 an essential membrane complex 140:168 an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity 140:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 12 theme essential 143:151 arg1 complex 123:129 The signal peptidase complex 102:129 The signal peptidase complex (SPC) 102:135 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 13 theme membrane 153:160 arg1 complex 162:168 an essential membrane complex 140:168 an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity 140:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 13 theme membrane 153:160 arg1 complex 123:129 The signal peptidase complex 102:129 The signal peptidase complex (SPC) 102:135 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 4 14 theme electron 626:633 arg1 proteomics 666:675 electron cryo-microscopy and structural proteomics 626:675 electron cryo-microscopy and structural proteomics 626:675 We determined the atomic structures of both paralogs using electron cryo-microscopy and structural proteomics. 34388369 2 15 theme molecular 400:408 arg1 details 410:416 the molecular details 396:416 the molecular details of SP recognition and removal 396:446 Although the determinants of this process have been established empirically, the molecular details of SP recognition and removal remain elusive. 34388369 4 16 theme paralogs 611:618 arg1 structures 592:601 the atomic structures 581:601 the atomic structures of both paralogs 581:618 We determined the atomic structures of both paralogs using electron cryo-microscopy and structural proteomics. 34388369 3 17 theme functional 511:520 arg1 paralogs 522:529 two functional paralogs 507:529 two functional paralogs with distinct proteolytic subunits 507:564 Here, we show that the human SPC exists in two functional paralogs with distinct proteolytic subunits. 34388369 0 18 theme human 17:21 arg1 complex 40:46 the human signal peptidase complex 13:46 the human signal peptidase complex 13:46 Structure of the human signal peptidase complex reveals the determinants for signal peptide cleavage. 34388369 4 19 theme atomic 585:590 arg1 structures 592:601 the atomic structures 581:601 the atomic structures of both paralogs 581:618 We determined the atomic structures of both paralogs using electron cryo-microscopy and structural proteomics. 34388369 1 20 theme endoplasmic 177:187 arg1 ER 200:201 ER 200:201 ER 200:201 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 20 theme endoplasmic 177:187 arg1 reticulum 189:197 the endoplasmic reticulum 173:197 the endoplasmic reticulum (ER) 173:202 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 0 21 theme peptidase 30:38 arg1 complex 40:46 the human signal peptidase complex 13:46 the human signal peptidase complex 13:46 Structure of the human signal peptidase complex reveals the determinants for signal peptide cleavage. 34388369 5 22 theme active 682:687 arg1 site 689:692 The active site 678:692 The active site 678:692 The active site is formed by a catalytic triad and abuts the ER membrane, where a transmembrane window collectively formed by all subunits locally thins the bilayer. 34388369 3 23 theme distinct 536:543 arg1 subunits 557:564 distinct proteolytic subunits 536:564 distinct proteolytic subunits 536:564 Here, we show that the human SPC exists in two functional paralogs with distinct proteolytic subunits. 34388369 2 24 theme recognition 424:434 arg1 details 410:416 the molecular details 396:416 the molecular details of SP recognition and removal 396:446 Although the determinants of this process have been established empirically, the molecular details of SP recognition and removal remain elusive. 34388369 0 25 theme signal 23:28 arg1 complex 40:46 the human signal peptidase complex 13:46 the human signal peptidase complex 13:46 Structure of the human signal peptidase complex reveals the determinants for signal peptide cleavage. 34388369 3 26 theme proteolytic 545:555 arg1 subunits 557:564 distinct proteolytic subunits 536:564 distinct proteolytic subunits 536:564 Here, we show that the human SPC exists in two functional paralogs with distinct proteolytic subunits. 34388369 5 27 theme transmembrane 760:772 arg1 window 774:779 a transmembrane window 758:779 a transmembrane window collectively formed by all subunits 758:815 The active site is formed by a catalytic triad and abuts the ER membrane, where a transmembrane window collectively formed by all subunits locally thins the bilayer. 34388369 2 28 theme SP 421:422 arg1 recognition 424:434 SP recognition 421:434 SP recognition 421:434 Although the determinants of this process have been established empirically, the molecular details of SP recognition and removal remain elusive. 34388369 1 29 theme large 251:255 arg1 variety 257:263 a large variety 249:263 a large variety of secretory pre-proteins with exquisite specificity 249:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 29 theme large 251:255 arg1 pre-proteins 278:289 secretory pre-proteins 268:289 secretory pre-proteins with exquisite specificity 268:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 30 theme signal 106:111 arg1 complex 123:129 The signal peptidase complex 102:129 The signal peptidase complex (SPC) 102:135 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 30 theme signal 106:111 arg1 complex 162:168 an essential membrane complex 140:168 an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity 140:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 30 theme signal 106:111 arg1 SPC 132:134 SPC 132:134 SPC 132:134 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 0 31 theme complex 40:46 arg1 Structure 0:8 Structure 0:8 Structure of the human signal peptidase complex 0:46 Structure of the human signal peptidase complex reveals the determinants for signal peptide cleavage. 34388369 2 32 theme process 353:359 arg1 determinants 332:343 the determinants 328:343 the determinants of this process 328:359 Although the determinants of this process have been established empirically, the molecular details of SP recognition and removal remain elusive. 34388369 1 33 theme peptidase 113:121 arg1 complex 123:129 The signal peptidase complex 102:129 The signal peptidase complex (SPC) 102:135 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 33 theme peptidase 113:121 arg1 complex 162:168 an essential membrane complex 140:168 an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity 140:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 33 theme peptidase 113:121 arg1 SPC 132:134 SPC 132:134 SPC 132:134 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 6 34 theme unique 894:899 arg1 architecture 901:912 this unique architecture 889:912 this unique architecture 889:912 Molecular dynamics simulations indicate that this unique architecture generates specificity for SPs based on the length of their hydrophobic segments. 34388369 1 35 from complex 162:168 arg1 ER 200:201 ER 200:201 ER 200:201 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 1 35 from complex 162:168 arg1 reticulum 189:197 the endoplasmic reticulum 173:197 the endoplasmic reticulum (ER) 173:202 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 5 36 theme catalytic 709:717 arg1 triad 719:723 a catalytic triad 707:723 a catalytic triad 707:723 The active site is formed by a catalytic triad and abuts the ER membrane, where a transmembrane window collectively formed by all subunits locally thins the bilayer. 34388369 6 37 theme segments 985:992 arg1 length 957:962 the length 953:962 the length of their hydrophobic segments 953:992 Molecular dynamics simulations indicate that this unique architecture generates specificity for SPs based on the length of their hydrophobic segments. 34388369 1 38 theme secretory 268:276 arg1 pre-proteins 278:289 secretory pre-proteins 268:289 secretory pre-proteins with exquisite specificity 268:316 The signal peptidase complex (SPC) is an essential membrane complex in the endoplasmic reticulum (ER), where it removes signal peptides (SPs) from a large variety of secretory pre-proteins with exquisite specificity. 34388369 5 39 theme ER 739:740 arg1 membrane 742:749 the ER membrane 735:749 the ER membrane 735:749 The active site is formed by a catalytic triad and abuts the ER membrane, where a transmembrane window collectively formed by all subunits locally thins the bilayer. 34388369 4 40 theme cryo-microscopy 635:649 arg1 proteomics 666:675 electron cryo-microscopy and structural proteomics 626:675 electron cryo-microscopy and structural proteomics 626:675 We determined the atomic structures of both paralogs using electron cryo-microscopy and structural proteomics. 34388369 6 41 theme hydrophobic 973:983 arg1 segments 985:992 their hydrophobic segments 967:992 their hydrophobic segments 967:992 Molecular dynamics simulations indicate that this unique architecture generates specificity for SPs based on the length of their hydrophobic segments. 12788922 9 0 used used 1158:1161 arg2 structure 1144:1152 The C1s structure 1136:1152 The C1s structure 1136:1152 The C1s structure was used to build a model of the C1r-C1s CUB1-EGF heterodimer, which in C1 connects C1r to C1s and mediates interaction with C1q. 12788922 10 1 theme structural 1286:1295 arg1 model 1297:1301 A structural model 1284:1301 A structural model of the C1q/C1r/C1s interface 1284:1330 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 3 2 theme N-terminal 277:286 arg1 elements 329:336 key elements 325:336 key elements of the C1 architecture 325:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 2 theme N-terminal 277:286 arg1 segments 297:304 The N-terminal CUB1-EGF segments 273:304 The N-terminal CUB1-EGF segments of C1r and C1s 273:319 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 3 theme C1r-C1s 403:409 arg1 association 411:421 Ca2+-dependent C1r-C1s association 388:421 Ca2+-dependent C1r-C1s association 388:421 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 7 4 theme distal 875:880 arg1 end 882:884 the distal end 871:884 the distal end of each CUB1 module 871:904 Unexpectedly, a second Ca2+ ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu45, Asp53, Asp98, and two water molecules. 12788922 3 5 theme C1s 317:319 arg1 elements 329:336 key elements 325:336 key elements of the C1 architecture 325:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 5 theme C1s 317:319 arg1 segments 297:304 The N-terminal CUB1-EGF segments 273:304 The N-terminal CUB1-EGF segments of C1r and C1s 273:319 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 5 6 theme factor 687:692 arg1 module 700:705 the epidermal growth factor (EGF) module 666:705 the epidermal growth factor (EGF) module of its counterpart 666:724 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 6 7 theme inter-monomer 801:813 arg1 interfaces 815:824 both the intra- and inter-monomer interfaces 781:824 interfaces 815:824 A Ca2+ ion is bound to each EGF module and stabilizes both the intra- and inter-monomer interfaces. 12788922 3 8 with interaction 427:437 arg1 C1q 444:446 C1q 444:446 C1q 444:446 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 6 9 theme Ca2+ 729:732 arg1 ion 734:736 A Ca2+ ion 727:736 A Ca2+ ion 727:736 A Ca2+ ion is bound to each EGF module and stabilizes both the intra- and inter-monomer interfaces. 12788922 9 10 theme C1s 1140:1142 arg1 structure 1144:1152 The C1s structure 1136:1152 The C1s structure 1136:1152 The C1s structure was used to build a model of the C1r-C1s CUB1-EGF heterodimer, which in C1 connects C1r to C1s and mediates interaction with C1q. 12788922 0 11 theme domain 48:53 arg1 structure 6:14 X-ray structure 0:14 X-ray structure of the Ca2+-binding interaction domain of C1s 0:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 0 12 theme interaction 36:46 arg1 C1s 58:60 C1s 58:60 C1s 58:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 0 12 theme interaction 36:46 arg1 domain 48:53 the Ca2+-binding interaction domain 19:53 the Ca2+-binding interaction domain of C1s 19:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 10 13 theme interface 1322:1330 arg1 model 1297:1301 A structural model 1284:1301 A structural model of the C1q/C1r/C1s interface 1284:1330 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 10 14 theme heterodimer 1465:1475 arg1 axis 1445:1448 the transversal axis 1429:1448 the transversal axis of the C1r-C1s heterodimer 1429:1475 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 10 15 theme transversal 1433:1443 arg1 axis 1445:1448 the transversal axis 1429:1448 the transversal axis of the C1r-C1s heterodimer 1429:1475 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 6 16 theme intra- 790:795 arg1 interfaces 815:824 both the intra- and inter-monomer interfaces 781:824 interfaces 815:824 A Ca2+ ion is bound to each EGF module and stabilizes both the intra- and inter-monomer interfaces. 12788922 10 17 theme C1q 1389:1391 arg1 helix 1380:1384 the rod-like collagen triple helix 1351:1384 the rod-like collagen triple helix of C1q 1351:1391 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 10 18 theme rod-like 1355:1362 arg1 helix 1380:1384 the rod-like collagen triple helix 1351:1384 the rod-like collagen triple helix of C1q 1351:1391 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 8 19 theme module 1121:1126 arg1 subset 1128:1133 a novel, Ca2+-binding CUB module subset 1095:1133 a novel, Ca2+-binding CUB module subset 1095:1133 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 0 20 theme X-ray 0:4 arg1 structure 6:14 X-ray structure 0:14 X-ray structure of the Ca2+-binding interaction domain of C1s 0:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 5 21 theme head-to-tail 577:588 arg1 homodimer 590:598 a head-to-tail homodimer 575:598 a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart 575:724 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 7 22 theme second 843:848 arg1 ion 855:857 a second Ca2+ ion 841:857 a second Ca2+ ion 841:857 Unexpectedly, a second Ca2+ ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu45, Asp53, Asp98, and two water molecules. 12788922 10 23 theme C1q/C1r/C1s 1310:1320 arg1 interface 1322:1330 the C1q/C1r/C1s interface 1306:1330 the C1q/C1r/C1s interface 1306:1330 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 2 24 dep proteases 220:228 arg1 C1s 238:240 C1s 238:240 C1s 238:240 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 2 24 dep proteases 220:228 arg1 C1r 230:232 C1r 230:232 C1r 230:232 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 8 25 theme repertoire 1073:1082 arg1 repertoire 1073:1082 the CUB repertoire 1065:1082 the CUB repertoire 1065:1082 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 8 25 theme repertoire 1073:1082 arg1 two-thirds 1051:1060 two-thirds 1051:1060 two-thirds 1051:1060 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 4 26 theme C1s 500:502 arg1 domain 490:495 the interaction domain 474:495 the interaction domain of C1s 474:502 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 4 26 theme C1s 500:502 arg1 C1s 500:502 C1s 500:502 C1s 500:502 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 8 27 dep novel 1097:1101 arg1 Ca2+-binding 1104:1115 Ca2+-binding 1104:1115 Ca2+-binding 1104:1115 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 7 28 theme Ca2+ 850:853 arg1 ion 855:857 a second Ca2+ ion 841:857 a second Ca2+ ion 841:857 Unexpectedly, a second Ca2+ ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu45, Asp53, Asp98, and two water molecules. 12788922 2 29 theme protein 260:266 arg1 C1q 268:270 a recognition protein C1q 246:270 a recognition protein C1q 246:270 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 3 30 theme key 325:327 arg1 elements 329:336 key elements 325:336 key elements of the C1 architecture 325:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 30 theme key 325:327 arg1 segments 297:304 The N-terminal CUB1-EGF segments 273:304 The N-terminal CUB1-EGF segments of C1r and C1s 273:319 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 9 31 theme CUB1-EGF 1195:1202 arg1 heterodimer 1204:1214 the C1r-C1s CUB1-EGF heterodimer 1183:1214 the C1r-C1s CUB1-EGF heterodimer 1183:1214 The C1s structure was used to build a model of the C1r-C1s CUB1-EGF heterodimer, which in C1 connects C1r to C1s and mediates interaction with C1q. 12788922 5 32 theme monomer 654:660 arg1 module 640:645 the CUB1 module 631:645 the CUB1 module of one monomer 631:660 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 5 32 theme monomer 654:660 arg1 module 700:705 the epidermal growth factor (EGF) module 666:705 the epidermal growth factor (EGF) module of its counterpart 666:724 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 2 33 theme recognition 248:258 arg1 C1q 268:270 a recognition protein C1q 246:270 a recognition protein C1q 246:270 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 3 34 theme architecture 348:359 arg1 elements 329:336 key elements 325:336 key elements of the C1 architecture 325:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 34 theme architecture 348:359 arg1 segments 297:304 The N-terminal CUB1-EGF segments 273:304 The N-terminal CUB1-EGF segments of C1r and C1s 273:319 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 9 35 theme heterodimer 1204:1214 arg1 model 1174:1178 a model 1172:1178 a model 1172:1178 The C1s structure was used to build a model of the C1r-C1s CUB1-EGF heterodimer, which in C1 connects C1r to C1s and mediates interaction with C1q. 12788922 3 36 with association 411:421 arg1 C1q 444:446 C1q 444:446 C1q 444:446 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 8 37 theme CUB 1117:1119 arg1 subset 1128:1133 a novel, Ca2+-binding CUB module subset 1095:1133 a novel, Ca2+-binding CUB module subset 1095:1133 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 3 38 theme Ca2+-dependent 388:401 arg1 association 411:421 Ca2+-dependent C1r-C1s association 388:421 Ca2+-dependent C1r-C1s association 388:421 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 0 39 theme Ca2+-binding 23:34 arg1 C1s 58:60 C1s 58:60 C1s 58:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 0 39 theme Ca2+-binding 23:34 arg1 domain 48:53 the Ca2+-binding interaction domain 19:53 the Ca2+-binding interaction domain of C1s 19:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 4 40 theme interaction 478:488 arg1 domain 490:495 the interaction domain 474:495 the interaction domain of C1s 474:502 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 4 40 theme interaction 478:488 arg1 C1s 500:502 C1s 500:502 C1s 500:502 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 5 41 theme counterpart 714:724 arg1 module 640:645 the CUB1 module 631:645 the CUB1 module of one monomer 631:660 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 5 41 theme counterpart 714:724 arg1 module 700:705 the epidermal growth factor (EGF) module 666:705 the epidermal growth factor (EGF) module of its counterpart 666:724 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 9 42 with interaction 1262:1272 arg1 C1q 1279:1281 C1q 1279:1281 C1q 1279:1281 The C1s structure was used to build a model of the C1r-C1s CUB1-EGF heterodimer, which in C1 connects C1r to C1s and mediates interaction with C1q. 12788922 3 43 theme C1r 309:311 arg1 elements 329:336 key elements 325:336 key elements of the C1 architecture 325:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 43 theme C1r 309:311 arg1 segments 297:304 The N-terminal CUB1-EGF segments 273:304 The N-terminal CUB1-EGF segments of C1r and C1s 273:319 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 8 44 theme novel 1097:1101 arg1 subset 1128:1133 a novel, Ca2+-binding CUB module subset 1095:1133 a novel, Ca2+-binding CUB module subset 1095:1133 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 5 45 theme epidermal 670:678 arg1 factor 687:692 epidermal growth factor 670:692 the epidermal growth factor (EGF) module of its counterpart 666:724 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 5 45 theme epidermal 670:678 arg1 EGF 695:697 EGF 695:697 EGF 695:697 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 10 46 theme collagen 1364:1371 arg1 helix 1380:1384 the rod-like collagen triple helix 1351:1384 the rod-like collagen triple helix of C1q 1351:1391 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 9 47 theme C1r-C1s 1187:1193 arg1 heterodimer 1204:1214 the C1r-C1s CUB1-EGF heterodimer 1183:1214 the C1r-C1s CUB1-EGF heterodimer 1183:1214 The C1s structure was used to build a model of the C1r-C1s CUB1-EGF heterodimer, which in C1 connects C1r to C1s and mediates interaction with C1q. 12788922 1 48 theme C1 97:98 arg1 complex 100:106 the C1 complex 93:106 the C1 complex of complement 93:120 Insights into the assembly of the C1 complex of complement. 12788922 3 49 theme C1 345:346 arg1 architecture 348:359 the C1 architecture 341:359 the C1 architecture 341:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 5 50 theme CUB1 635:638 arg1 module 640:645 the CUB1 module 631:645 the CUB1 module of one monomer 631:660 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 2 51 theme complement 178:187 arg1 pathway 167:173 the classical pathway 153:173 the classical pathway of complement 153:187 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 7 52 theme water 971:975 arg1 molecules 977:985 two water molecules 967:985 two water molecules 967:985 Unexpectedly, a second Ca2+ ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu45, Asp53, Asp98, and two water molecules. 12788922 7 53 theme module 899:904 arg1 end 882:884 the distal end 871:884 the distal end of each CUB1 module 871:904 Unexpectedly, a second Ca2+ ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu45, Asp53, Asp98, and two water molecules. 12788922 1 54 theme complex 100:106 arg1 assembly 81:88 the assembly 77:88 the assembly of the C1 complex of complement 77:120 Insights into the assembly of the C1 complex of complement. 12788922 10 55 theme C1r-C1s 1457:1463 arg1 heterodimer 1465:1475 the C1r-C1s heterodimer 1453:1475 the C1r-C1s heterodimer 1453:1475 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 4 56 theme domain 490:495 arg1 structure 461:469 The crystal structure 449:469 The crystal structure of the interaction domain of C1s 449:502 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 8 57 theme acidic 994:999 arg1 residues 1001:1008 These acidic residues 988:1008 These acidic residues 988:1008 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 6 58 theme EGF 755:757 arg1 module 759:764 each EGF module 750:764 each EGF module 750:764 A Ca2+ ion is bound to each EGF module and stabilizes both the intra- and inter-monomer interfaces. 12788922 1 59 theme complement 111:120 arg1 complex 100:106 the C1 complex 93:106 the C1 complex of complement 93:120 Insights into the assembly of the C1 complex of complement. 12788922 4 60 theme A 539:539 arg1 resolution 541:550 1.5 A resolution 535:550 1.5 A resolution 535:550 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 3 61 theme CUB1-EGF 288:295 arg1 elements 329:336 key elements 325:336 key elements of the C1 architecture 325:359 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 3 61 theme CUB1-EGF 288:295 arg1 segments 297:304 The N-terminal CUB1-EGF segments 273:304 The N-terminal CUB1-EGF segments of C1r and C1s 273:319 The N-terminal CUB1-EGF segments of C1r and C1s are key elements of the C1 architecture, because they mediate both Ca2+-dependent C1r-C1s association and interaction with C1q. 12788922 2 62 theme classical 157:165 arg1 pathway 167:173 the classical pathway 153:173 the classical pathway of complement 153:187 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 2 63 theme modular 212:218 arg1 proteases 220:228 two modular proteases C1r and C1s 208:240 two modular proteases C1r and C1s 208:240 C1, the complex that triggers the classical pathway of complement, is assembled from two modular proteases C1r and C1s and a recognition protein C1q. 12788922 7 64 theme CUB1 894:897 arg1 module 899:904 each CUB1 module 889:904 each CUB1 module 889:904 Unexpectedly, a second Ca2+ ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu45, Asp53, Asp98, and two water molecules. 12788922 10 65 theme triple 1373:1378 arg1 helix 1380:1384 the rod-like collagen triple helix 1351:1384 the rod-like collagen triple helix of C1q 1351:1391 A structural model of the C1q/C1r/C1s interface is proposed, where the rod-like collagen triple helix of C1q is accommodated into a groove along the transversal axis of the C1r-C1s heterodimer. 12788922 0 66 theme C1s 58:60 arg1 C1s 58:60 C1s 58:60 C1s 58:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 0 66 theme C1s 58:60 arg1 domain 48:53 the Ca2+-binding interaction domain 19:53 the Ca2+-binding interaction domain of C1s 19:60 X-ray structure of the Ca2+-binding interaction domain of C1s. 12788922 4 67 theme crystal 453:459 arg1 structure 461:469 The crystal structure 449:469 The crystal structure of the interaction domain of C1s 449:502 The crystal structure of the interaction domain of C1s has been solved and refined to 1.5 A resolution. 12788922 8 68 theme CUB 1069:1071 arg1 repertoire 1073:1082 the CUB repertoire 1065:1082 the CUB repertoire 1065:1082 These acidic residues and Tyr17 are conserved in approximately two-thirds of the CUB repertoire and define a novel, Ca2+-binding CUB module subset. 12788922 5 69 theme growth 680:685 arg1 factor 687:692 epidermal growth factor 670:692 the epidermal growth factor (EGF) module of its counterpart 666:724 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 12788922 5 69 theme growth 680:685 arg1 EGF 695:697 EGF 695:697 EGF 695:697 The structure reveals a head-to-tail homodimer involving interactions between the CUB1 module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 25053408 11 0 theme loop 1662:1665 arg1 structure 1667:1675 a novel reentrant loop structure 1644:1675 a novel reentrant loop structure 1644:1675 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 10 1 theme Pemetrexed 1405:1414 arg1 pretreatment 1416:1427 Pemetrexed pretreatment 1405:1427 Pemetrexed pretreatment 1405:1427 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 6 2 theme TMD2-3 875:880 arg1 loop 882:885 the TMD2-3 loop 871:885 the TMD2-3 loop in a Cys-less PCFT background 871:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 11 3 theme 2-3 1596:1598 arg1 aqueous-accessible 1615:1632 aqueous-accessible 1615:1632 aqueous-accessible 1615:1632 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 11 3 theme 2-3 1596:1598 arg1 domain 1605:1610 the TMD 2-3 loop domain 1588:1610 the TMD 2-3 loop domain 1588:1610 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 0 4 theme Substituted 0:10 arg1 accessibility 21:33 Substituted cysteine accessibility 0:33 Substituted cysteine accessibility 0:33 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 1 5 theme proton-coupled 183:196 arg1 symporter 243:251 a folate-proton symporter 227:251 a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions 227:385 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 1 5 theme proton-coupled 183:196 arg1 PCFT 218:221 PCFT 218:221 PCFT 218:221 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 1 5 theme proton-coupled 183:196 arg1 transporter 205:215 The proton-coupled folate transporter 179:215 The proton-coupled folate transporter (PCFT) 179:222 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 12 6 from Residues 1678:1685 arg1 TMD2 1690:1693 TMD2 1690:1693 TMD2 1690:1693 Residues in TMD2 form an aqueous transmembrane pathway for folate substrates, and Gly-93 and Phe-94 may contribute to a substrate binding domain. 25053408 8 7 theme pulldown 1109:1116 arg1 assays 1118:1123 pulldown assays 1109:1123 pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1109:1190 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 6 8 theme PCFT-null 939:947 arg1 cells 954:958 PCFT-null HeLa cells 939:958 PCFT-null HeLa cells 939:958 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 4 9 theme solute 639:644 arg1 carriers 646:653 other solute carriers 633:653 other solute carriers 633:653 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 9 10 from residues 1268:1275 arg1 domain 1305:1310 the TMD2-3 loop domain 1289:1310 the TMD2-3 loop domain 1289:1310 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 9 10 from residues 1268:1275 arg1 TMD2 1280:1283 TMD2 1280:1283 TMD2 1280:1283 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 8 11 theme 2-aminoethyl 1151:1162 arg1 methanethiosulfonate-biotin 1164:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 4 12 theme domain 705:710 arg1 part 670:673 part 670:673 part of the PCFT substrate binding domain 670:710 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 0 13 theme proton-coupled 144:157 arg1 transporter 166:176 the human proton-coupled folate transporter 134:176 the human proton-coupled folate transporter 134:176 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 6 14 theme Cys-less 892:899 arg1 background 906:915 a Cys-less PCFT background 890:915 a Cys-less PCFT background 890:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 0 15 from role 99:102 arg1 transporter 166:176 the human proton-coupled folate transporter 134:176 the human proton-coupled folate transporter 134:176 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 0 16 theme reentrant 69:77 arg1 loop 79:82 a novel transmembrane 2-3 reentrant loop 43:82 a novel transmembrane 2-3 reentrant loop 43:82 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 12 17 theme transmembrane 1711:1723 arg1 pathway 1725:1731 an aqueous transmembrane pathway 1700:1731 an aqueous transmembrane pathway for folate substrates 1700:1753 Residues in TMD2 form an aqueous transmembrane pathway for folate substrates, and Gly-93 and Phe-94 may contribute to a substrate binding domain. 25053408 7 18 from pH 1063:1064 arg1 active 1023:1028 active 1023:1028 active 1023:1028 All 33 mutants were detected by Western blotting, and 28 were active for [(3)H]methotrexate uptake at pH 5.5. 25053408 8 19 with assays 1118:1123 arg1 methanethiosulfonate-biotin 1164:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 4 20 theme substrate 687:695 arg1 domain 705:710 the PCFT substrate binding domain 678:710 the PCFT substrate binding domain 678:710 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 1 21 theme microenvironment 359:374 arg1 conditions 376:385 acidic microenvironment conditions 352:385 acidic microenvironment conditions 352:385 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 10 22 theme mutants 1461:1467 arg1 biotinylation 1439:1451 biotinylation 1439:1451 biotinylation of TMD2 mutants G93C and F94C 1439:1481 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 13 23 theme substrate 1952:1960 arg1 binding 1962:1968 substrate binding 1952:1968 substrate binding 1952:1968 Characterization of PCFT structure is essential to understanding the transport mechanism including the critical determinants of substrate binding. 25053408 0 24 theme transmembrane 108:120 arg1 domain 122:127 transmembrane domain 2 108:129 transmembrane domain 2 108:129 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 10 25 dep mutants 1461:1467 arg1 G93C 1469:1472 G93C 1469:1472 G93C 1469:1472 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 10 25 dep mutants 1461:1467 arg1 mutants 1461:1467 TMD2 mutants 1456:1467 TMD2 mutants G93C and F94C 1456:1481 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 10 25 dep mutants 1461:1467 arg1 F94C 1478:1481 F94C 1478:1481 F94C 1478:1481 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 5 26 theme PCFT 781:784 arg1 residues 763:770 the seven cysteine (Cys) residues 738:770 the seven cysteine (Cys) residues of human PCFT to serine 738:794 In this study we mutated the seven cysteine (Cys) residues of human PCFT to serine, creating Cys-less PCFT. 25053408 5 26 theme PCFT 781:784 arg1 PCFT 781:784 human PCFT 775:784 human PCFT to serine 775:794 In this study we mutated the seven cysteine (Cys) residues of human PCFT to serine, creating Cys-less PCFT. 25053408 9 27 theme loop 1300:1303 arg1 domain 1305:1310 the TMD2-3 loop domain 1289:1310 the TMD2-3 loop domain 1289:1310 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 2 28 theme loop 440:443 arg1 domain 445:450 the loop domain 436:450 the loop domain between transmembrane domains (TMDs) 2 and 3 436:495 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 1 29 theme cytotoxic 314:322 arg1 antifolates 324:334 cytotoxic antifolates 314:334 cytotoxic antifolates 314:334 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 6 30 theme Thirty-three 821:832 arg1 mutants 845:851 Thirty-three single-Cys mutants 821:851 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background 821:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 9 31 theme 2-aminoethyl 1325:1336 arg1 methanethiosulfonate-biotin 1338:1364 2-aminoethyl methanethiosulfonate-biotin 1325:1364 2-aminoethyl methanethiosulfonate-biotin 1325:1364 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 0 32 theme transmembrane 51:63 arg1 loop 79:82 a novel transmembrane 2-3 reentrant loop 43:82 a novel transmembrane 2-3 reentrant loop 43:82 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 10 33 theme transport 1543:1551 arg1 activity 1553:1560 methotrexate transport activity 1530:1560 methotrexate transport activity 1530:1560 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 9 34 theme aqueous 1380:1386 arg1 accessibilities 1388:1402 aqueous accessibilities 1380:1402 aqueous accessibilities 1380:1402 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 6 35 from loop 882:885 arg1 background 906:915 a Cys-less PCFT background 890:915 a Cys-less PCFT background 890:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 11 36 theme reentrant 1652:1660 arg1 structure 1667:1675 a novel reentrant loop structure 1644:1675 a novel reentrant loop structure 1644:1675 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 13 37 theme structure 1849:1857 arg1 Characterization 1824:1839 Characterization 1824:1839 Characterization of PCFT structure 1824:1857 Characterization of PCFT structure is essential to understanding the transport mechanism including the critical determinants of substrate binding. 25053408 5 38 theme cysteine 748:755 arg1 residues 763:770 the seven cysteine (Cys) residues 738:770 the seven cysteine (Cys) residues of human PCFT to serine 738:794 In this study we mutated the seven cysteine (Cys) residues of human PCFT to serine, creating Cys-less PCFT. 25053408 5 38 theme cysteine 748:755 arg1 PCFT 781:784 human PCFT 775:784 human PCFT to serine 775:794 In this study we mutated the seven cysteine (Cys) residues of human PCFT to serine, creating Cys-less PCFT. 25053408 7 39 theme methotrexate 1040:1051 arg1 H 1038:1038 3)H 1036:1038 3)H 1036:1038 All 33 mutants were detected by Western blotting, and 28 were active for [(3)H]methotrexate uptake at pH 5.5. 25053408 7 39 theme methotrexate 1040:1051 arg1 uptake 1053:1058 [(3)H]methotrexate uptake 1034:1058 [(3)H]methotrexate uptake 1034:1058 All 33 mutants were detected by Western blotting, and 28 were active for [(3)H]methotrexate uptake at pH 5.5. 25053408 2 40 theme Predicted 388:396 arg1 models 407:412 Predicted topology models 388:412 Predicted topology models for PCFT 388:421 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 9 41 theme Multiple 1259:1266 arg1 residues 1268:1275 Multiple residues 1259:1275 Multiple residues in TMD2 and the TMD2-3 loop domain 1259:1310 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 6 42 from TMD2 862:865 arg1 background 906:915 a Cys-less PCFT background 890:915 a Cys-less PCFT background 890:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 1 43 theme solid 273:277 arg1 tumors 279:284 solid tumors 273:284 solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions 273:385 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 12 44 theme substrate 1798:1806 arg1 domain 1816:1821 a substrate binding domain 1796:1821 a substrate binding domain 1796:1821 Residues in TMD2 form an aqueous transmembrane pathway for folate substrates, and Gly-93 and Phe-94 may contribute to a substrate binding domain. 25053408 8 45 theme active 1079:1084 arg1 residues 1086:1093 the active residues 1075:1093 the active residues 1075:1093 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 7 46 dep H 1038:1038 arg1 3 1036:1036 3 1036:1036 3 1036:1036 All 33 mutants were detected by Western blotting, and 28 were active for [(3)H]methotrexate uptake at pH 5.5. 25053408 11 47 theme loop 1600:1603 arg1 aqueous-accessible 1615:1632 aqueous-accessible 1615:1632 aqueous-accessible 1615:1632 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 11 47 theme loop 1600:1603 arg1 domain 1605:1610 the TMD 2-3 loop domain 1588:1610 the TMD 2-3 loop domain 1588:1610 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 1 48 theme folate 198:203 arg1 symporter 243:251 a folate-proton symporter 227:251 a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions 227:385 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 1 48 theme folate 198:203 arg1 PCFT 218:221 PCFT 218:221 PCFT 218:221 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 1 48 theme folate 198:203 arg1 transporter 205:215 The proton-coupled folate transporter 179:215 The proton-coupled folate transporter (PCFT) 179:222 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 6 49 theme HeLa 949:952 arg1 cells 954:958 PCFT-null HeLa cells 939:958 PCFT-null HeLa cells 939:958 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 8 50 theme streptavidin 1196:1207 arg1 beads 1209:1213 streptavidin beads 1196:1213 streptavidin beads 1196:1213 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 11 51 theme TMD 1592:1594 arg1 aqueous-accessible 1615:1632 aqueous-accessible 1615:1632 aqueous-accessible 1615:1632 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 11 51 theme TMD 1592:1594 arg1 domain 1605:1610 the TMD 2-3 loop domain 1588:1610 the TMD 2-3 loop domain 1588:1610 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 3 52 theme TMD2-3 567:572 arg1 loop 574:577 the TMD2-3 loop 563:577 the TMD2-3 loop 563:577 Mutations involving Asp-109 or Arg-113 in the TMD2-3 loop result in loss of activity. 25053408 4 53 theme other 633:637 arg1 carriers 646:653 other solute carriers 633:653 other solute carriers 633:653 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 0 54 theme human 138:142 arg1 transporter 166:176 the human proton-coupled folate transporter 134:176 the human proton-coupled folate transporter 134:176 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 3 55 theme activity 597:604 arg1 loss 589:592 loss 589:592 loss of activity 589:604 Mutations involving Asp-109 or Arg-113 in the TMD2-3 loop result in loss of activity. 25053408 5 56 theme Cys-less 806:813 arg1 PCFT 815:818 Cys-less PCFT 806:818 Cys-less PCFT 806:818 In this study we mutated the seven cysteine (Cys) residues of human PCFT to serine, creating Cys-less PCFT. 25053408 0 57 from loop 79:82 arg1 transporter 166:176 the human proton-coupled folate transporter 134:176 the human proton-coupled folate transporter 134:176 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 0 58 theme folate 159:164 arg1 transporter 166:176 the human proton-coupled folate transporter 134:176 the human proton-coupled folate transporter 134:176 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 8 59 theme membrane-impermeable 1130:1149 arg1 methanethiosulfonate-biotin 1164:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 6 60 theme PCFT 901:904 arg1 background 906:915 a Cys-less PCFT background 890:915 a Cys-less PCFT background 890:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 4 61 theme binding 697:703 arg1 domain 705:710 the PCFT substrate binding domain 678:710 the PCFT substrate binding domain 678:710 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 13 62 theme binding 1962:1968 arg1 determinants 1936:1947 the critical determinants 1923:1947 the critical determinants of substrate binding 1923:1968 Characterization of PCFT structure is essential to understanding the transport mechanism including the critical determinants of substrate binding. 25053408 7 63 theme Western 993:999 arg1 blotting 1001:1008 Western blotting 993:1008 Western blotting 993:1008 All 33 mutants were detected by Western blotting, and 28 were active for [(3)H]methotrexate uptake at pH 5.5. 25053408 12 64 theme folate 1737:1742 arg1 substrates 1744:1753 folate substrates 1737:1753 folate substrates 1737:1753 Residues in TMD2 form an aqueous transmembrane pathway for folate substrates, and Gly-93 and Phe-94 may contribute to a substrate binding domain. 25053408 13 65 theme critical 1927:1934 arg1 determinants 1936:1947 the critical determinants 1923:1947 the critical determinants of substrate binding 1923:1968 Characterization of PCFT structure is essential to understanding the transport mechanism including the critical determinants of substrate binding. 25053408 0 66 theme functional 88:97 arg1 role 99:102 functional role 88:102 functional role for transmembrane domain 2 in the human proton-coupled folate transporter 88:176 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 1 67 theme acidic 352:357 arg1 conditions 376:385 acidic microenvironment conditions 352:385 acidic microenvironment conditions 352:385 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 12 68 theme aqueous 1703:1709 arg1 pathway 1725:1731 an aqueous transmembrane pathway 1700:1731 an aqueous transmembrane pathway for folate substrates 1700:1753 Residues in TMD2 form an aqueous transmembrane pathway for folate substrates, and Gly-93 and Phe-94 may contribute to a substrate binding domain. 25053408 4 69 theme PCFT 682:685 arg1 domain 705:710 the PCFT substrate binding domain 678:710 the PCFT substrate binding domain 678:710 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 10 70 theme TMD2 1456:1459 arg1 G93C 1469:1472 G93C 1469:1472 G93C 1469:1472 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 10 70 theme TMD2 1456:1459 arg1 mutants 1461:1467 TMD2 mutants 1456:1467 TMD2 mutants G93C and F94C 1456:1481 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 10 70 theme TMD2 1456:1459 arg1 F94C 1478:1481 F94C 1478:1481 F94C 1478:1481 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 13 71 theme transport 1893:1901 arg1 mechanism 1903:1911 the transport mechanism 1889:1911 the transport mechanism including the critical determinants of substrate binding 1889:1968 Characterization of PCFT structure is essential to understanding the transport mechanism including the critical determinants of substrate binding. 25053408 2 72 dep domains 474:480 arg1 2 489:489 2 489:489 2 489:489 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 2 72 dep domains 474:480 arg1 3 495:495 3 495:495 3 495:495 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 5 73 theme human 775:779 arg1 PCFT 781:784 human PCFT 775:784 human PCFT to serine 775:794 In this study we mutated the seven cysteine (Cys) residues of human PCFT to serine, creating Cys-less PCFT. 25053408 9 74 theme TMD2-3 1293:1298 arg1 domain 1305:1310 the TMD2-3 loop domain 1289:1310 the TMD2-3 loop domain 1289:1310 Multiple residues in TMD2 and the TMD2-3 loop domain reacted with 2-aminoethyl methanethiosulfonate-biotin, establishing aqueous accessibilities. 25053408 2 75 theme transmembrane 460:472 arg1 TMDs 483:486 TMDs 483:486 TMDs 483:486 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 2 75 theme transmembrane 460:472 arg1 domains 474:480 transmembrane domains 460:480 transmembrane domains (TMDs) 2 and 3 460:495 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 10 76 theme residues 1511:1518 arg1 biotinylation 1488:1500 biotinylation 1488:1500 biotinylation of these residues 1488:1518 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 0 77 theme novel 45:49 arg1 loop 79:82 a novel transmembrane 2-3 reentrant loop 43:82 a novel transmembrane 2-3 reentrant loop 43:82 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 6 78 theme single-Cys 834:843 arg1 mutants 845:851 Thirty-three single-Cys mutants 821:851 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background 821:915 Thirty-three single-Cys mutants spanning TMD2 and the TMD2-3 loop in a Cys-less PCFT background were transfected into PCFT-null HeLa cells. 25053408 0 79 theme 2-3 65:67 arg1 loop 79:82 a novel transmembrane 2-3 reentrant loop 43:82 a novel transmembrane 2-3 reentrant loop 43:82 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 25053408 4 80 theme structural 610:619 arg1 homology 621:628 structural homology 610:628 structural homology to other solute carriers 610:653 By structural homology to other solute carriers, TMD2 may form part of the PCFT substrate binding domain. 25053408 10 81 theme methotrexate 1530:1541 arg1 activity 1553:1560 methotrexate transport activity 1530:1560 methotrexate transport activity 1530:1560 Pemetrexed pretreatment inhibited biotinylation of TMD2 mutants G93C and F94C, and biotinylation of these residues inhibited methotrexate transport activity. 25053408 11 82 theme novel 1646:1650 arg1 structure 1667:1675 a novel reentrant loop structure 1644:1675 a novel reentrant loop structure 1644:1675 Our results suggest that the TMD 2-3 loop domain is aqueous-accessible and forms a novel reentrant loop structure. 25053408 2 83 theme topology 398:405 arg1 models 407:412 Predicted topology models 388:412 Predicted topology models for PCFT 388:421 Predicted topology models for PCFT suggest that the loop domain between transmembrane domains (TMDs) 2 and 3 resides in the cytosol. 25053408 1 84 theme folate-proton 229:241 arg1 symporter 243:251 a folate-proton symporter 227:251 a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions 227:385 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 1 84 theme folate-proton 229:241 arg1 transporter 205:215 The proton-coupled folate transporter 179:215 The proton-coupled folate transporter (PCFT) 179:222 The proton-coupled folate transporter (PCFT) is a folate-proton symporter highly expressed in solid tumors that can selectively target cytotoxic antifolates to tumors under acidic microenvironment conditions. 25053408 12 85 theme binding 1808:1814 arg1 domain 1816:1821 a substrate binding domain 1796:1821 a substrate binding domain 1796:1821 Residues in TMD2 form an aqueous transmembrane pathway for folate substrates, and Gly-93 and Phe-94 may contribute to a substrate binding domain. 25053408 13 86 theme PCFT 1844:1847 arg1 structure 1849:1857 PCFT structure 1844:1857 PCFT structure 1844:1857 Characterization of PCFT structure is essential to understanding the transport mechanism including the critical determinants of substrate binding. 25053408 8 87 with beads 1209:1213 arg1 methanethiosulfonate-biotin 1164:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin 1130:1190 For the active residues, we performed pulldown assays with membrane-impermeable 2-aminoethyl methanethiosulfonate-biotin and streptavidin beads to determine their aqueous-accessibilities. 25053408 0 88 theme cysteine 12:19 arg1 accessibility 21:33 Substituted cysteine accessibility 0:33 Substituted cysteine accessibility 0:33 Substituted cysteine accessibility reveals a novel transmembrane 2-3 reentrant loop and functional role for transmembrane domain 2 in the human proton-coupled folate transporter. 30301806 6 0 theme substrate- 924:933 arg1 site 956:959 a stable substrate- or inhibitor-binding site 915:959 a stable substrate- or inhibitor-binding site 915:959 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 1 1 theme animal 165:170 arg1 models 172:177 animal models 165:177 animal models 165:177 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 6 2 theme stable 917:922 arg1 site 956:959 a stable substrate- or inhibitor-binding site 915:959 a stable substrate- or inhibitor-binding site 915:959 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 6 3 from pair 1024:1027 arg1 enzyme 1059:1064 the enzyme 1055:1064 the enzyme 1055:1064 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 2 4 from levels 333:338 arg1 cells 350:354 immune cells 343:354 immune cells 343:354 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 4 5 theme various 699:705 arg1 states 718:723 various activation states 699:723 various activation states 699:723 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 3 6 theme Pharmacological 357:371 arg1 inhibitors 373:382 Pharmacological inhibitors 357:382 Pharmacological inhibitors of NAAA activity 357:399 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 6 7 from helices 1044:1050 arg1 enzyme 1059:1064 the enzyme 1055:1064 the enzyme 1055:1064 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 4 8 theme molecular 609:617 arg1 mechanism 619:627 the molecular mechanism 605:627 the molecular mechanism of action of NAAA 605:645 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 6 9 theme active 1154:1159 arg1 site 1161:1164 the active site 1150:1164 the active site that accommodates the ligand's acyl chain 1150:1206 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 1 10 from inflammation 149:160 arg1 humans 186:191 humans 186:191 humans 186:191 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 1 10 from inflammation 149:160 arg1 models 172:177 animal models 165:177 animal models 165:177 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 4 11 theme activation 707:716 arg1 states 718:723 various activation states 699:723 various activation states 699:723 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 6 12 theme pair 1024:1027 arg1 interaction 1007:1017 the interaction 1003:1017 the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes 1003:1085 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 6 13 theme cavity 1138:1143 arg1 creation 1105:1112 the creation 1101:1112 the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain 1101:1206 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 5 14 theme buried 855:860 arg1 NAAA 877:880 NAAA 877:880 NAAA 877:880 Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. 30301806 5 14 theme buried 855:860 arg1 site 869:872 the otherwise buried active site 841:872 the otherwise buried active site of NAAA 841:880 Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. 30301806 3 15 theme rodent 458:463 arg1 models 465:470 rodent models 458:470 rodent models 458:470 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 0 16 theme immunoregulatory 41:56 arg1 NAAA 66:69 the immunoregulatory amidase NAAA 37:69 the immunoregulatory amidase NAAA 37:69 Molecular mechanism of activation of the immunoregulatory amidase NAAA. 30301806 1 17 from pain 140:143 arg1 humans 186:191 humans 186:191 humans 186:191 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 1 17 from pain 140:143 arg1 models 172:177 animal models 165:177 animal models 165:177 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 4 18 from structures 670:679 arg1 states 718:723 various activation states 699:723 various activation states 699:723 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 4 18 from structures 670:679 arg1 complex 732:738 complex 732:738 complex with several ligands, including both a covalent and a reversible inhibitor 732:813 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 4 19 theme crystal 662:668 arg1 structures 670:679 crystal structures 662:679 crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor 662:813 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 3 20 theme activity 392:399 arg1 inhibitors 373:382 Pharmacological inhibitors 357:382 Pharmacological inhibitors of NAAA activity 357:399 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 4 21 theme several 745:751 arg1 ligands 753:759 several ligands 745:759 several ligands 745:759 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 4 21 theme several 745:751 arg1 inhibitor 805:813 a reversible inhibitor 792:813 a reversible inhibitor 792:813 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 4 21 theme several 745:751 arg1 covalent 779:786 covalent 779:786 covalent 779:786 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 0 22 theme Molecular 0:8 arg1 mechanism 10:18 Molecular mechanism 0:18 Molecular mechanism of activation of the immunoregulatory amidase NAAA 0:69 Molecular mechanism of activation of the immunoregulatory amidase NAAA. 30301806 1 23 theme bioactive 99:107 arg1 Palmitoylethanolamide 72:92 Palmitoylethanolamide 72:92 Palmitoylethanolamide 72:92 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 1 23 theme bioactive 99:107 arg1 lipid 109:113 a bioactive lipid 97:113 a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans 97:191 Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. 30301806 6 24 theme lipid 1071:1075 arg1 membranes 1077:1085 lipid membranes 1071:1085 lipid membranes 1071:1085 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 0 25 theme activation 23:32 arg1 mechanism 10:18 Molecular mechanism 0:18 Molecular mechanism of activation of the immunoregulatory amidase NAAA 0:69 Molecular mechanism of activation of the immunoregulatory amidase NAAA. 30301806 6 26 theme acyl 1197:1200 arg1 chain 1202:1206 the ligand's acyl chain 1184:1206 the ligand's acyl chain 1184:1206 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 6 27 from enzyme 1059:1064 arg1 pair 1024:1027 a pair 1022:1027 a pair of hydrophobic helices in the enzyme 1022:1064 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 3 28 theme NAAA 387:390 arg1 activity 392:399 NAAA activity 387:399 NAAA activity 387:399 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 2 29 theme cysteine 296:303 arg1 hydrolase 305:313 a cysteine hydrolase 294:313 a cysteine hydrolase expressed at high levels in immune cells 294:354 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 2 29 theme cysteine 296:303 arg1 amidase 278:284 N-acylethanolamine acid amidase 254:284 N-acylethanolamine acid amidase (NAAA) 254:291 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 4 30 theme NAAA 642:645 arg1 action 632:637 action 632:637 action of NAAA 632:645 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 6 31 theme hydrophobic 1126:1136 arg1 cavity 1138:1143 a linear hydrophobic cavity 1117:1143 a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain 1117:1206 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 5 32 theme active 862:867 arg1 NAAA 877:880 NAAA 877:880 NAAA 877:880 Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. 30301806 5 32 theme active 862:867 arg1 site 869:872 the otherwise buried active site 841:872 the otherwise buried active site of NAAA 841:880 Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. 30301806 3 33 theme antiinflammatory 430:445 arg1 effects 447:453 profound analgesic and antiinflammatory effects 407:453 profound analgesic and antiinflammatory effects 407:453 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 6 34 theme inhibitor-binding 938:954 arg1 site 956:959 a stable substrate- or inhibitor-binding site 915:959 a stable substrate- or inhibitor-binding site 915:959 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 3 35 theme profound 407:414 arg1 effects 447:453 profound analgesic and antiinflammatory effects 407:453 profound analgesic and antiinflammatory effects 407:453 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 0 36 theme NAAA 66:69 arg1 activation 23:32 activation 23:32 activation of the immunoregulatory amidase NAAA 23:69 Molecular mechanism of activation of the immunoregulatory amidase NAAA. 30301806 2 37 theme immune 343:348 arg1 cells 350:354 immune cells 343:354 immune cells 343:354 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 3 38 theme analgesic 416:424 arg1 effects 447:453 profound analgesic and antiinflammatory effects 407:453 profound analgesic and antiinflammatory effects 407:453 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 6 39 theme site 956:959 arg1 Formation 902:910 Formation 902:910 Formation of a stable substrate- or inhibitor-binding site 902:959 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 3 40 theme potential 503:511 arg1 target 513:518 a potential target 501:518 a potential target for therapeutic drug discovery 501:549 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 4 41 theme reversible 794:803 arg1 inhibitor 805:813 a reversible inhibitor 792:813 a reversible inhibitor 792:813 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 6 42 with interaction 1007:1017 arg1 membranes 1077:1085 lipid membranes 1071:1085 lipid membranes 1071:1085 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 4 43 theme enzyme 689:694 arg1 structures 670:679 crystal structures 662:679 crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor 662:813 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 6 44 theme linear 1119:1124 arg1 cavity 1138:1143 a linear hydrophobic cavity 1117:1143 a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain 1117:1206 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 6 45 theme helices 1044:1050 arg1 pair 1024:1027 a pair 1022:1027 a pair of hydrophobic helices in the enzyme 1022:1064 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 2 46 theme signaling 198:206 arg1 activity 208:215 Its signaling activity 194:215 Its signaling activity 194:215 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 2 47 theme high 328:331 arg1 levels 333:338 high levels 328:338 high levels in immune cells 328:354 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 6 48 theme hydrophobic 1032:1042 arg1 helices 1044:1050 hydrophobic helices 1032:1050 hydrophobic helices in the enzyme 1032:1064 Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain. 30301806 2 49 theme acid 273:276 arg1 hydrolase 305:313 a cysteine hydrolase 294:313 a cysteine hydrolase expressed at high levels in immune cells 294:354 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 2 49 theme acid 273:276 arg1 NAAA 287:290 NAAA 287:290 NAAA 287:290 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 2 49 theme acid 273:276 arg1 amidase 278:284 N-acylethanolamine acid amidase 254:284 N-acylethanolamine acid amidase (NAAA) 254:291 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 3 50 theme therapeutic 524:534 arg1 discovery 541:549 therapeutic drug discovery 524:549 therapeutic drug discovery 524:549 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 5 51 theme NAAA 877:880 arg1 NAAA 877:880 NAAA 877:880 NAAA 877:880 Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. 30301806 5 51 theme NAAA 877:880 arg1 site 869:872 the otherwise buried active site 841:872 the otherwise buried active site of NAAA 841:880 Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. 30301806 0 52 theme amidase 58:64 arg1 NAAA 66:69 the immunoregulatory amidase NAAA 37:69 the immunoregulatory amidase NAAA 37:69 Molecular mechanism of activation of the immunoregulatory amidase NAAA. 30301806 4 53 with complex 732:738 arg1 ligands 753:759 several ligands 745:759 several ligands 745:759 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 4 53 with complex 732:738 arg1 inhibitor 805:813 a reversible inhibitor 792:813 a reversible inhibitor 792:813 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 4 53 with complex 732:738 arg1 covalent 779:786 covalent 779:786 covalent 779:786 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 30301806 2 54 theme N-acylethanolamine 254:271 arg1 hydrolase 305:313 a cysteine hydrolase 294:313 a cysteine hydrolase expressed at high levels in immune cells 294:354 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 2 54 theme N-acylethanolamine 254:271 arg1 NAAA 287:290 NAAA 287:290 NAAA 287:290 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 2 54 theme N-acylethanolamine 254:271 arg1 amidase 278:284 N-acylethanolamine acid amidase 254:284 N-acylethanolamine acid amidase (NAAA) 254:291 Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. 30301806 3 55 theme drug 536:539 arg1 discovery 541:549 therapeutic drug discovery 524:549 therapeutic drug discovery 524:549 Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. 30301806 4 56 theme action 632:637 arg1 mechanism 619:627 the molecular mechanism 605:627 the molecular mechanism of action of NAAA 605:645 To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. 11707436 3 0 theme hexosaminidase 665:678 arg1 activity 680:687 hexosaminidase activity 665:687 hexosaminidase activity 665:687 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 6 1 theme incomplete 1227:1236 arg1 sulfate 1247:1253 incomplete dermatan sulfate 1227:1253 incomplete dermatan sulfate degradation 1227:1265 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 1 2 theme Hex 303:305 arg1 alphabeta 310:318 alphabeta 310:318 alphabeta 310:318 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 2 theme Hex 303:305 arg1 A 307:307 Hex A 303:307 Hex A (alphabeta) 303:319 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 8 3 theme lysosomal 1682:1690 arg1 phospholipid 1700:1711 the lysosomal anionic phospholipid 1678:1711 the lysosomal anionic phospholipid bis(monoacylglycero)phosphate 1678:1741 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 7 4 theme amphiphilic 1431:1441 arg1 SM2 1551:1553 the sulfated glycosphingolipid SM2 1520:1553 the sulfated glycosphingolipid SM2 1520:1553 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 4 theme amphiphilic 1431:1441 arg1 glycoconjugates 1443:1457 water-soluble and amphiphilic glycoconjugates 1413:1457 water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2 1413:1553 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 4 theme amphiphilic 1431:1441 arg1 substrates 1480:1489 artificial substrates 1469:1489 artificial substrates 1469:1489 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 4 theme amphiphilic 1431:1441 arg1 fragments 1505:1513 sulfated GAG fragments 1492:1513 sulfated GAG fragments 1492:1513 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 2 5 theme Hex 529:531 arg1 S 533:533 labile Hex S 522:533 labile Hex S 522:533 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 3 6 theme S 720:720 arg1 Nat 882:884 Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat 716:884 Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat 716:884 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 7 7 theme water-soluble 1413:1425 arg1 SM2 1551:1553 the sulfated glycosphingolipid SM2 1520:1553 the sulfated glycosphingolipid SM2 1520:1553 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 7 theme water-soluble 1413:1425 arg1 glycoconjugates 1443:1457 water-soluble and amphiphilic glycoconjugates 1413:1457 water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2 1413:1553 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 7 theme water-soluble 1413:1425 arg1 substrates 1480:1489 artificial substrates 1469:1489 artificial substrates 1469:1489 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 7 theme water-soluble 1413:1425 arg1 fragments 1505:1513 sulfated GAG fragments 1492:1513 sulfated GAG fragments 1492:1513 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 3 8 from activity 680:687 arg1 deficient 652:660 deficient 652:660 deficient 652:660 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 8 9 theme membrane-bound 1570:1583 arg1 SM2 1585:1587 membrane-bound SM2 1570:1587 membrane-bound SM2 1570:1587 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 3 10 dep N. 763:764 arg1 M. 773:774 M. 773:774 M. 773:774 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 3 10 dep N. 763:764 arg1 L. 776:777 L. 776:777 L. 776:777 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 3 11 theme anionic 595:601 arg1 oligosaccharides 603:618 anionic oligosaccharides 595:618 anionic oligosaccharides 595:618 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 8 12 theme recombinant 1596:1606 arg1 S 1612:1612 the recombinant Hex S 1592:1612 the recombinant Hex S 1592:1612 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 7 13 theme glycosphingolipid 1533:1549 arg1 SM2 1551:1553 the sulfated glycosphingolipid SM2 1520:1553 the sulfated glycosphingolipid SM2 1520:1553 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 7 14 theme Hex 1386:1388 arg1 S 1390:1390 recombinant Hex S 1374:1390 recombinant Hex S 1374:1390 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 5 15 theme knockout 1092:1099 arg1 mice 1101:1104 the double knockout mice 1081:1104 the double knockout mice 1081:1104 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 3 16 dep S 720:720 arg1 L 871:871 Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L 723:871 Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L 723:871 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 6 17 theme degradation 1351:1361 arg1 fragments 1329:1337 fragments 1329:1337 fragments of N-glycan degradation 1329:1361 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 6 17 theme degradation 1351:1361 arg1 oligosaccharides 1295:1310 the neutral storage oligosaccharides 1275:1310 the neutral storage oligosaccharides 1275:1310 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 1 18 link linked 265:270 arg1 beta 291:294 beta 291:294 beta 291:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 18 link linked 265:270 arg1 alpha 281:285 alpha 281:285 alpha 281:285 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 18 link linked 265:270 arg1 subunits 272:279 two noncovalently linked subunits 247:279 two noncovalently linked subunits alpha and beta 247:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 19 theme Hex 344:346 arg1 alphaalpha 351:360 alphaalpha 351:360 alphaalpha 351:360 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 19 theme Hex 344:346 arg1 S 348:348 Hex S 344:348 Hex S (alphaalpha) 344:361 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 2 20 theme physiological 492:504 arg1 significance 506:517 the physiological significance 488:517 the physiological significance of labile Hex S 488:533 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 0 21 theme terminal 113:120 arg1 residues 167:174 terminal beta-glycosidically bound N-acetylhexosamine residues 113:174 terminal beta-glycosidically bound N-acetylhexosamine residues 113:174 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 3 22 theme R. 868:869 arg1 L 871:871 Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L 723:871 Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L 723:871 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 7 23 theme GAG 1501:1503 arg1 fragments 1505:1513 sulfated GAG fragments 1492:1513 sulfated GAG fragments 1492:1513 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 3 24 from accumulation 579:590 arg1 mice 700:703 mice 700:703 mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat 700:884 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 3 24 from accumulation 579:590 arg1 mice 639:642 double knockout mice 623:642 double knockout mice totally deficient in hexosaminidase activity 623:687 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 0 25 theme bound 142:146 arg1 residues 167:174 terminal beta-glycosidically bound N-acetylhexosamine residues 113:174 terminal beta-glycosidically bound N-acetylhexosamine residues 113:174 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 8 26 theme GM2 1652:1654 arg1 protein 1666:1672 the GM2 activator protein 1648:1672 the GM2 activator protein 1648:1672 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 0 27 theme lysosomal 75:83 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 0 28 theme Physiological 0:12 arg1 substrates 14:23 Physiological substrates 0:23 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 0:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 7 29 theme artificial 1469:1478 arg1 substrates 1480:1489 artificial substrates 1469:1489 artificial substrates 1469:1489 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 2 30 theme B 392:392 arg1 role 374:377 the role 370:377 the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates 370:459 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 2 31 theme neutral 437:443 arg1 glycoconjugates 445:459 several anionic and neutral glycoconjugates 417:459 several anionic and neutral glycoconjugates 417:459 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 3 32 theme knockout 630:637 arg1 mice 639:642 double knockout mice 623:642 double knockout mice totally deficient in hexosaminidase activity 623:687 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 2 33 theme A 386:386 arg1 role 374:377 the role 370:377 the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates 370:459 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 6 34 theme anionic 1180:1186 arg1 glycans 1188:1194 the anionic glycans 1176:1194 the anionic glycans 1176:1194 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 6 34 theme anionic 1180:1186 arg1 products 1215:1222 products 1215:1222 products of incomplete dermatan sulfate degradation 1215:1265 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 0 35 theme lysosomal 35:43 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 2 36 theme anionic 425:431 arg1 glycoconjugates 445:459 several anionic and neutral glycoconjugates 417:459 several anionic and neutral glycoconjugates 417:459 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 0 37 theme Human 69:73 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 0 38 theme -hexosaminidase 50:64 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 5 39 theme physiological 984:996 arg1 substrates 998:1007 physiological substrates 984:1007 physiological substrates of Hex S 984:1016 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 6 40 theme degradation 1255:1265 arg1 glycans 1188:1194 the anionic glycans 1176:1194 the anionic glycans 1176:1194 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 6 40 theme degradation 1255:1265 arg1 products 1215:1222 products 1215:1222 products of incomplete dermatan sulfate degradation 1215:1265 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 3 41 from deficient 652:660 arg1 activity 680:687 hexosaminidase activity 665:687 hexosaminidase activity 665:687 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 6 42 theme dermatan 1238:1245 arg1 sulfate 1247:1253 incomplete dermatan sulfate 1227:1253 incomplete dermatan sulfate degradation 1227:1265 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 5 43 theme Hex 965:967 arg1 specificity 950:960 the substrate specificity 936:960 the substrate specificity of Hex S. To identify physiological substrates of Hex S 936:1016 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 8 44 theme anionic 1692:1698 arg1 phospholipid 1700:1711 the lysosomal anionic phospholipid 1678:1711 the lysosomal anionic phospholipid bis(monoacylglycero)phosphate 1678:1741 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 8 45 theme Hex 1608:1610 arg1 S 1612:1612 the recombinant Hex S 1592:1612 the recombinant Hex S 1592:1612 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 3 46 theme Hex 716:718 arg1 S 720:720 Hex S 716:720 Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat 716:884 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 3 46 theme Hex 716:718 arg1 1996 876:879 1996 876:879 1996 876:879 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 8 47 theme bis 1713:1715 arg1 phosphate 1733:1741 bis(monoacylglycero)phosphate 1713:1741 the lysosomal anionic phospholipid bis(monoacylglycero)phosphate 1678:1741 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 1 48 theme different 216:224 arg1 isozymes 226:233 Three different isozymes 210:233 Three different isozymes composed of two noncovalently linked subunits alpha and beta 210:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 8 49 dep phospholipid 1700:1711 arg1 phosphate 1733:1741 bis(monoacylglycero)phosphate 1713:1741 the lysosomal anionic phospholipid bis(monoacylglycero)phosphate 1678:1741 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 8 50 theme monoacylglycero 1717:1731 arg1 phosphate 1733:1741 bis(monoacylglycero)phosphate 1713:1741 the lysosomal anionic phospholipid bis(monoacylglycero)phosphate 1678:1741 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 3 51 theme K. 853:854 arg1 L 871:871 Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L 723:871 Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L 723:871 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 2 52 theme labile 522:527 arg1 S 533:533 labile Hex S 522:533 labile Hex S 522:533 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 6 53 theme storage 1287:1293 arg1 fragments 1329:1337 fragments 1329:1337 fragments of N-glycan degradation 1329:1361 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 6 53 theme storage 1287:1293 arg1 oligosaccharides 1295:1310 the neutral storage oligosaccharides 1275:1310 the neutral storage oligosaccharides 1275:1310 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 1 54 theme linked 265:270 arg1 beta 291:294 beta 291:294 beta 291:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 54 theme linked 265:270 arg1 alpha 281:285 alpha 281:285 alpha 281:285 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 54 theme linked 265:270 arg1 subunits 272:279 two noncovalently linked subunits 247:279 two noncovalently linked subunits alpha and beta 247:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 0 55 theme glycoconjugates 193:207 arg1 number 183:188 a number 181:188 a number of glycoconjugates 181:207 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 5 56 theme neutral 1031:1037 arg1 oligosaccharides 1039:1054 anionic and neutral oligosaccharides 1019:1054 anionic and neutral oligosaccharides excreted in the urine of the double knockout mice 1019:1104 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 8 57 theme SM2 1585:1587 arg1 Hydrolysis 1556:1565 Hydrolysis 1556:1565 Hydrolysis of membrane-bound SM2 by the recombinant Hex S 1556:1612 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 7 58 theme recombinant 1374:1384 arg1 S 1390:1390 recombinant Hex S 1374:1390 recombinant Hex S 1374:1390 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 2 59 theme S 533:533 arg1 significance 506:517 the physiological significance 488:517 the physiological significance of labile Hex S 488:533 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 5 60 dep Hex 965:967 arg1 S. 969:970 Hex S. 965:970 Hex S. 965:970 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 6 61 theme glycosidase 1154:1164 arg1 digestion 1166:1174 glycosidase digestion 1154:1174 glycosidase digestion 1154:1174 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 3 62 theme striking 570:577 arg1 accumulation 579:590 the striking accumulation 566:590 the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat 566:884 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 5 63 theme S 1016:1016 arg1 substrates 998:1007 physiological substrates 984:1007 physiological substrates of Hex S 984:1016 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 6 64 theme N-glycan 1342:1349 arg1 degradation 1351:1361 N-glycan degradation 1342:1361 N-glycan degradation 1342:1361 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 7 65 theme sulfated 1524:1531 arg1 SM2 1551:1553 the sulfated glycosphingolipid SM2 1520:1553 the sulfated glycosphingolipid SM2 1520:1553 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 2 66 theme glycoconjugates 445:459 arg1 degradation 402:412 the degradation 398:412 the degradation of several anionic and neutral glycoconjugates 398:459 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 5 67 theme anionic 1019:1025 arg1 oligosaccharides 1039:1054 anionic and neutral oligosaccharides 1019:1054 anionic and neutral oligosaccharides excreted in the urine of the double knockout mice 1019:1104 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 5 68 theme double 1085:1090 arg1 mice 1101:1104 the double knockout mice 1081:1104 the double knockout mice 1081:1104 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 0 69 theme N-acetylhexosamine 148:165 arg1 residues 167:174 terminal beta-glycosidically bound N-acetylhexosamine residues 113:174 terminal beta-glycosidically bound N-acetylhexosamine residues 113:174 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 5 70 theme mice 1101:1104 arg1 urine 1072:1076 the urine 1068:1076 the urine of the double knockout mice 1068:1104 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 7 71 theme sulfated 1492:1499 arg1 fragments 1505:1513 sulfated GAG fragments 1492:1513 sulfated GAG fragments 1492:1513 In vitro, recombinant Hex S was highly active on water-soluble and amphiphilic glycoconjugates including artificial substrates, sulfated GAG fragments, and the sulfated glycosphingolipid SM2. 11707436 0 72 theme human 29:33 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 5 73 theme substrate 940:948 arg1 specificity 950:960 the substrate specificity 936:960 the substrate specificity of Hex S. To identify physiological substrates of Hex S 936:1016 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 8 74 theme activator 1656:1664 arg1 protein 1666:1672 the GM2 activator protein 1648:1672 the GM2 activator protein 1648:1672 Hydrolysis of membrane-bound SM2 by the recombinant Hex S was synergistically stimulated by the GM2 activator protein and the lysosomal anionic phospholipid bis(monoacylglycero)phosphate. 11707436 5 75 dep 14 894:895 arg1 348-352 898:904 348-352 898:904 348-352 898:904 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 11707436 3 76 theme double 623:628 arg1 mice 639:642 double knockout mice 623:642 double knockout mice totally deficient in hexosaminidase activity 623:687 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 0 77 theme beta 45:48 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 1 78 theme Hex 322:324 arg1 betabeta 329:336 betabeta 329:336 betabeta 329:336 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 78 theme Hex 322:324 arg1 B 326:326 Hex B 322:326 Hex B (betabeta) 322:337 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 3 79 theme oligosaccharides 603:618 arg1 accumulation 579:590 the striking accumulation 566:590 the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat 566:884 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 2 80 theme Hex 382:384 arg1 A 386:386 Hex A 382:386 Hex A 382:386 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 0 81 theme S. 66:67 arg1 beta-hexosaminidases 85:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases 29:104 Physiological substrates for human lysosomal beta -hexosaminidase S. Human lysosomal beta-hexosaminidases remove terminal beta-glycosidically bound N-acetylhexosamine residues from a number of glycoconjugates. 11707436 6 82 dep identified 1201:1210 arg1 Using 1134:1138 Using 1134:1138 Using ESI-MS/MS and glycosidase digestion 1134:1174 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 2 83 theme several 417:423 arg1 glycoconjugates 445:459 several anionic and neutral glycoconjugates 417:459 several anionic and neutral glycoconjugates 417:459 While the role of Hex A and B for the degradation of several anionic and neutral glycoconjugates has been well established, the physiological significance of labile Hex S has remained unclear. 11707436 3 84 theme deficient 652:660 arg1 mice 639:642 double knockout mice 623:642 double knockout mice totally deficient in hexosaminidase activity 623:687 However, the striking accumulation of anionic oligosaccharides in double knockout mice totally deficient in hexosaminidase activity but not in mice expressing Hex S (Sango, K., McDonald, M. P., Crawley, J. N., Mack, M. L., Tifft, C.J., Skop, E., Starr, C. M., Hoffmann, A., Sandhoff, K., Suzuki, K., and Proia, R. L., (1996) Nat. 11707436 6 85 theme neutral 1279:1285 arg1 fragments 1329:1337 fragments 1329:1337 fragments of N-glycan degradation 1329:1361 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 6 85 theme neutral 1279:1285 arg1 oligosaccharides 1295:1310 the neutral storage oligosaccharides 1275:1310 the neutral storage oligosaccharides 1275:1310 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 1 86 dep subunits 272:279 arg1 beta 291:294 beta 291:294 beta 291:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 86 dep subunits 272:279 arg1 alpha 281:285 alpha 281:285 alpha 281:285 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 1 86 dep subunits 272:279 arg1 subunits 272:279 two noncovalently linked subunits 247:279 two noncovalently linked subunits alpha and beta 247:294 Three different isozymes composed of two noncovalently linked subunits alpha and beta exist: Hex A (alphabeta), Hex B (betabeta), and Hex S (alphaalpha). 11707436 6 87 theme sulfate 1247:1253 arg1 degradation 1255:1265 incomplete dermatan sulfate degradation 1227:1265 incomplete dermatan sulfate degradation 1227:1265 Using ESI-MS/MS and glycosidase digestion the anionic glycans were identified as products of incomplete dermatan sulfate degradation whereas the neutral storage oligosaccharides were found to be fragments of N-glycan degradation. 11707436 5 88 theme Hex 1012:1014 arg1 S 1016:1016 Hex S 1012:1016 Hex S 1012:1016 14, 348-352) prompted us to reinvestigate the substrate specificity of Hex S. To identify physiological substrates of Hex S, anionic and neutral oligosaccharides excreted in the urine of the double knockout mice were isolated and analyzed. 4055801 17 0 theme major 2020:2024 arg1 parallel 2031:2038 at least one major fold parallel 2007:2038 at least one major fold parallel 2007:2038 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 7 1 theme = 802:802 arg1 Mr 799:800 Mr 799:800 Mr = 28,000 799:809 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 14 2 located found 1798:1802 arg1 periphery 1811:1819 the periphery 1807:1819 the periphery of the torus and base 1807:1841 The oligosaccharide positions were found on the periphery of the torus and base. 4055801 14 2 located found 1798:1802 arg2 positions 1783:1791 The oligosaccharide positions 1763:1791 The oligosaccharide positions 1763:1791 The oligosaccharide positions were found on the periphery of the torus and base. 4055801 13 3 theme concanavalin 1723:1734 arg1 A 1736:1736 concanavalin A 1723:1736 concanavalin A 1723:1736 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 7 4 dep amino- 849:854 arg1 the 845:847 the 845:847 the 845:847 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 17 5 theme tubule 2067:2072 arg1 axis 2055:2058 the central axis 2043:2058 the central axis of the tubule 2043:2072 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 2 6 theme visible 269:275 arg1 crevice 261:267 a crevice 259:267 a crevice visible on one face 259:287 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 8 7 theme C9b 1162:1164 arg1 resistance 1083:1092 the resistance 1079:1092 the resistance of poly(C9) to denaturation 1079:1120 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 7 theme C9b 1162:1164 arg1 feature 1138:1144 a collective feature 1125:1144 a collective feature of both C9a and C9b 1125:1164 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 7 8 dep = 824:824 arg1 38,000 826:831 38,000 826:831 38,000 826:831 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 5 9 theme poly 531:534 arg1 micrographs 516:526 Electron micrographs 507:526 Electron micrographs of poly(C9) 507:538 Electron micrographs of poly(C9) indicate that the torus is formed by radial strands of polypeptide. 4055801 14 10 theme base 1838:1841 arg1 periphery 1811:1819 the periphery 1807:1819 the periphery of the torus and base 1807:1841 The oligosaccharide positions were found on the periphery of the torus and base. 4055801 9 11 theme immunoelectron 1235:1248 arg1 microscopy 1250:1259 immunoelectron microscopy 1235:1259 immunoelectron microscopy 1235:1259 The C9a and C9b polypeptide regions have been mapped on poly(C9) by immunoelectron microscopy. 4055801 5 12 theme radial 577:582 arg1 strands 584:590 radial strands 577:590 radial strands of polypeptide 577:605 Electron micrographs of poly(C9) indicate that the torus is formed by radial strands of polypeptide. 4055801 12 13 theme polypeptide 1546:1556 arg1 segments 1527:1534 the C9a and C9b segments 1511:1534 the C9a and C9b segments of the C9 polypeptide 1511:1556 Thus, the C9a and C9b segments of the C9 polypeptide are not clearly segregated in poly(C9). 4055801 7 14 theme protein 894:900 arg1 segments 878:885 carboxyl-terminal segments 860:885 carboxyl-terminal segments 860:885 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 14 theme protein 894:900 arg1 fragments 783:791 two single-chain polypeptide fragments 754:791 two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000) 754:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 14 theme protein 894:900 arg1 amino- 849:854 amino- 849:854 amino- 849:854 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 11 15 theme C9b 1379:1381 arg1 epitopes 1383:1390 C9b epitopes 1379:1390 C9b epitopes 1379:1390 C9b epitopes are concentrated predominantly about the torus and base, but were rarely observed on the midsection of poly(C9). 4055801 14 16 theme torus 1828:1832 arg1 periphery 1811:1819 the periphery 1807:1819 the periphery of the torus and base 1807:1841 The oligosaccharide positions were found on the periphery of the torus and base. 4055801 17 17 theme polypeptide 1979:1989 arg1 chain 1991:1995 the polypeptide chain 1975:1995 the polypeptide chain 1975:1995 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 11 18 theme poly 1495:1498 arg1 midsection 1481:1490 the midsection 1477:1490 the midsection of poly(C9) 1477:1502 C9b epitopes are concentrated predominantly about the torus and base, but were rarely observed on the midsection of poly(C9). 4055801 12 19 theme C9a 1515:1517 arg1 segments 1527:1534 the C9a and C9b segments 1511:1534 the C9a and C9b segments of the C9 polypeptide 1511:1556 Thus, the C9a and C9b segments of the C9 polypeptide are not clearly segregated in poly(C9). 4055801 11 20 located observed 1465:1472 arg2 epitopes 1383:1390 C9b epitopes 1379:1390 C9b epitopes 1379:1390 C9b epitopes are concentrated predominantly about the torus and base, but were rarely observed on the midsection of poly(C9). 4055801 11 20 located observed 1465:1472 arg1 midsection 1481:1490 the midsection 1477:1490 the midsection of poly(C9) 1477:1502 C9b epitopes are concentrated predominantly about the torus and base, but were rarely observed on the midsection of poly(C9). 4055801 2 21 theme ellipsoid 221:229 arg1 appearance 231:240 an ellipsoid appearance 218:240 an ellipsoid appearance (70 X 50 A) 218:252 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 2 21 theme ellipsoid 221:229 arg1 A 251:251 70 X 50 A 243:251 70 X 50 A 243:251 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 7 22 dep C9b 816:818 arg1 Mr 821:822 Mr 821:822 Mr = 38,000 821:831 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 10 23 theme cylinder 1369:1376 arg1 midsection 1342:1351 the midsection 1338:1351 the midsection of the poly(C9) cylinder 1338:1376 Determinants for the C9a region were observed about the torus, base, and on the midsection of the poly(C9) cylinder. 4055801 8 24 theme protein 941:947 arg1 C9a 950:952 C9a 950:952 C9a 950:952 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 24 theme protein 941:947 arg1 form 929:932 The cleaved form 917:932 The cleaved form of the protein 917:947 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 7 25 theme single-chain 758:769 arg1 C9b 816:818 C9b 816:818 C9b (Mr = 38,000) 816:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 25 theme single-chain 758:769 arg1 C9a 794:796 C9a 794:796 C9a (Mr = 28,000) 794:810 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 25 theme single-chain 758:769 arg1 fragments 783:791 two single-chain polypeptide fragments 754:791 two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000) 754:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 25 theme single-chain 758:769 arg1 amino- 849:854 amino- 849:854 amino- 849:854 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 13 26 from locations 1602:1610 arg1 C9 1645:1646 C9 1645:1646 C9 1645:1646 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 13 26 from locations 1602:1610 arg1 poly 1640:1643 poly 1640:1643 poly(C9) 1640:1647 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 4 27 theme 90 443:444 arg1 A-inner 446:452 A-inner 446:452 A-inner 446:452 Poly(C9) is a cylinder (150 A-outer diameters and 90 A-inner diameter) rimmed by a torus (46-A thick) on one end. 4055801 6 28 theme structure 702:710 arg1 axis 678:681 the central axis 666:681 the central axis of the cylindrical structure 666:710 Each subunit of poly(C9) is apparently tilted relative to the central axis of the cylindrical structure. 4055801 12 29 theme C9b 1523:1525 arg1 segments 1527:1534 the C9a and C9b segments 1511:1534 the C9a and C9b segments of the C9 polypeptide 1511:1556 Thus, the C9a and C9b segments of the C9 polypeptide are not clearly segregated in poly(C9). 4055801 7 30 theme polypeptide 771:781 arg1 C9b 816:818 C9b 816:818 C9b (Mr = 38,000) 816:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 30 theme polypeptide 771:781 arg1 C9a 794:796 C9a 794:796 C9a (Mr = 28,000) 794:810 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 30 theme polypeptide 771:781 arg1 fragments 783:791 two single-chain polypeptide fragments 754:791 two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000) 754:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 30 theme polypeptide 771:781 arg1 amino- 849:854 amino- 849:854 amino- 849:854 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 5 31 theme Electron 507:514 arg1 micrographs 516:526 Electron micrographs 507:526 Electron micrographs of poly(C9) 507:538 Electron micrographs of poly(C9) indicate that the torus is formed by radial strands of polypeptide. 4055801 6 32 theme poly 624:627 arg1 subunit 613:619 Each subunit 608:619 Each subunit of poly(C9) 608:631 Each subunit of poly(C9) is apparently tilted relative to the central axis of the cylindrical structure. 4055801 6 33 theme central 670:676 arg1 axis 678:681 the central axis 666:681 the central axis of the cylindrical structure 666:710 Each subunit of poly(C9) is apparently tilted relative to the central axis of the cylindrical structure. 4055801 13 34 with complex 1710:1716 arg1 A 1736:1736 concanavalin A 1723:1736 concanavalin A 1723:1736 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 1 35 theme transmission 141:152 arg1 microscopy 163:172 transmission electron microscopy 141:172 transmission electron microscopy 141:172 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 4 36 theme A-inner 446:452 arg1 diameter 454:461 90 A-inner diameter 443:461 90 A-inner diameter 443:461 Poly(C9) is a cylinder (150 A-outer diameters and 90 A-inner diameter) rimmed by a torus (46-A thick) on one end. 4055801 7 37 dep = 802:802 arg1 28,000 804:809 28,000 804:809 28,000 804:809 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 1 38 theme electron 154:161 arg1 microscopy 163:172 transmission electron microscopy 141:172 transmission electron microscopy 141:172 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 10 39 theme C9a 1283:1285 arg1 region 1287:1292 the C9a region 1279:1292 the C9a region 1279:1292 Determinants for the C9a region were observed about the torus, base, and on the midsection of the poly(C9) cylinder. 4055801 9 40 theme C9b 1179:1181 arg1 regions 1195:1201 The C9a and C9b polypeptide regions 1167:1201 The C9a and C9b polypeptide regions 1167:1201 The C9a and C9b polypeptide regions have been mapped on poly(C9) by immunoelectron microscopy. 4055801 8 41 theme poly 1097:1100 arg1 resistance 1083:1092 the resistance 1079:1092 the resistance of poly(C9) to denaturation 1079:1120 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 41 theme poly 1097:1100 arg1 feature 1138:1144 a collective feature 1125:1144 a collective feature of both C9a and C9b 1125:1164 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 6 42 theme cylindrical 690:700 arg1 structure 702:710 the cylindrical structure 686:710 the cylindrical structure 686:710 Each subunit of poly(C9) is apparently tilted relative to the central axis of the cylindrical structure. 4055801 1 43 theme human 78:82 arg1 C9 105:106 human complement component C9 78:106 human complement component C9 78:106 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 14 44 theme oligosaccharide 1767:1781 arg1 positions 1783:1791 The oligosaccharide positions 1763:1791 The oligosaccharide positions 1763:1791 The oligosaccharide positions were found on the periphery of the torus and base. 4055801 13 45 theme electron 1668:1675 arg1 microscopy 1677:1686 electron microscopy 1668:1686 electron microscopy 1668:1686 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 3 46 theme hollow 327:332 arg1 structures 342:351 hollow tubular structures 327:351 hollow tubular structures consisting of 12-16 monomeric subunits 327:390 C9 polymerizes spontaneously to form hollow tubular structures consisting of 12-16 monomeric subunits. 4055801 8 47 theme sulfate-resistant 1035:1051 arg1 C9 1058:1059 C9 1058:1059 C9 1058:1059 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 47 theme sulfate-resistant 1035:1051 arg1 poly 1053:1056 sodium dodecyl sulfate-resistant poly 1020:1056 sodium dodecyl sulfate-resistant poly(C9) 1020:1060 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 4 48 dep torus 476:480 arg1 thick 488:492 thick 488:492 thick 488:492 Poly(C9) is a cylinder (150 A-outer diameters and 90 A-inner diameter) rimmed by a torus (46-A thick) on one end. 4055801 8 49 theme suitable 992:999 arg1 conditions 1001:1010 suitable conditions 992:1010 suitable conditions 992:1010 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 3 50 theme monomeric 373:381 arg1 subunits 383:390 12-16 monomeric subunits 367:390 12-16 monomeric subunits 367:390 C9 polymerizes spontaneously to form hollow tubular structures consisting of 12-16 monomeric subunits. 4055801 0 51 theme component 31:39 arg1 C9 41:42 component C9 31:42 component C9 31:42 The architecture of complement component C9 and poly(C9). 4055801 17 52 theme poly 1966:1969 arg1 subunit 1955:1961 a subunit 1953:1961 a subunit of poly(C9) 1953:1973 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 3 53 theme tubular 334:340 arg1 structures 342:351 hollow tubular structures 327:351 hollow tubular structures consisting of 12-16 monomeric subunits 327:390 C9 polymerizes spontaneously to form hollow tubular structures consisting of 12-16 monomeric subunits. 4055801 8 54 theme cleaved 921:927 arg1 C9a 950:952 C9a 950:952 C9a 950:952 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 54 theme cleaved 921:927 arg1 form 929:932 The cleaved form 917:932 The cleaved form of the protein 917:947 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 12 55 theme C9 1543:1544 arg1 polypeptide 1546:1556 the C9 polypeptide 1539:1556 the C9 polypeptide 1539:1556 Thus, the C9a and C9b segments of the C9 polypeptide are not clearly segregated in poly(C9). 4055801 9 56 theme polypeptide 1183:1193 arg1 regions 1195:1201 The C9a and C9b polypeptide regions 1167:1201 The C9a and C9b polypeptide regions 1167:1201 The C9a and C9b polypeptide regions have been mapped on poly(C9) by immunoelectron microscopy. 4055801 4 57 theme A-outer 421:427 arg1 diameters 429:437 150 A-outer diameters 417:437 150 A-outer diameters 417:437 Poly(C9) is a cylinder (150 A-outer diameters and 90 A-inner diameter) rimmed by a torus (46-A thick) on one end. 4055801 8 58 theme collective 1127:1136 arg1 resistance 1083:1092 the resistance 1079:1092 the resistance of poly(C9) to denaturation 1079:1120 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 58 theme collective 1127:1136 arg1 feature 1138:1144 a collective feature 1125:1144 a collective feature of both C9a and C9b 1125:1164 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 17 59 dep form 2002:2005 arg1 form 1948:1951 form 1948:1951 To form a subunit of poly(C9) 1945:1973 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 1 60 theme complement 84:93 arg1 C9 105:106 human complement component C9 78:106 human complement component C9 78:106 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 4 61 dep cylinder 407:414 arg1 diameter 454:461 90 A-inner diameter 443:461 90 A-inner diameter 443:461 Poly(C9) is a cylinder (150 A-outer diameters and 90 A-inner diameter) rimmed by a torus (46-A thick) on one end. 4055801 4 61 dep cylinder 407:414 arg1 diameters 429:437 150 A-outer diameters 417:437 150 A-outer diameters 417:437 Poly(C9) is a cylinder (150 A-outer diameters and 90 A-inner diameter) rimmed by a torus (46-A thick) on one end. 4055801 2 62 theme Monomeric 175:183 arg1 Mr 196:197 Mr = 66,000 196:206 Mr = 66,000 196:206 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 2 62 theme Monomeric 175:183 arg1 C9 192:193 Monomeric native C9 175:193 Monomeric native C9 (Mr = 66,000) 175:207 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 2 63 theme native 185:190 arg1 Mr 196:197 Mr = 66,000 196:206 Mr = 66,000 196:206 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 2 63 theme native 185:190 arg1 C9 192:193 Monomeric native C9 175:193 Monomeric native C9 (Mr = 66,000) 175:207 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 13 64 theme oligosaccharide 1615:1629 arg1 units 1631:1635 oligosaccharide units 1615:1635 oligosaccharide units 1615:1635 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 1 65 theme component 95:103 arg1 C9 105:106 human complement component C9 78:106 human complement component C9 78:106 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 7 66 dep fragments 783:791 arg1 C9b 816:818 C9b 816:818 C9b (Mr = 38,000) 816:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 66 dep fragments 783:791 arg1 C9a 794:796 C9a 794:796 C9a (Mr = 28,000) 794:810 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 66 dep fragments 783:791 arg1 fragments 783:791 two single-chain polypeptide fragments 754:791 two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000) 754:832 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 67 theme = 824:824 arg1 Mr 821:822 Mr 821:822 Mr = 38,000 821:831 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 16 68 theme major 1924:1928 arg1 rearrangement 1930:1942 a major rearrangement 1922:1942 a major rearrangement 1922:1942 Polymerization involves a major rearrangement. 4055801 13 69 theme complex 1710:1716 arg1 labeling 1694:1701 labeling 1694:1701 labeling of the complex with concanavalin A bound to colloidal gold 1694:1760 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 1 70 theme C9 105:106 arg1 architecture 62:73 The architecture 58:73 The architecture of human complement component C9 and poly(C9) 58:119 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 13 71 theme colloidal 1747:1755 arg1 gold 1757:1760 colloidal gold 1747:1760 colloidal gold 1747:1760 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 13 72 theme units 1631:1635 arg1 locations 1602:1610 The locations 1598:1610 The locations of oligosaccharide units on poly(C9) 1598:1647 The locations of oligosaccharide units on poly(C9) were visualized by electron microscopy after labeling of the complex with concanavalin A bound to colloidal gold. 4055801 10 73 theme poly 1360:1363 arg1 cylinder 1369:1376 the poly(C9) cylinder 1356:1376 the poly(C9) cylinder 1356:1376 Determinants for the C9a region were observed about the torus, base, and on the midsection of the poly(C9) cylinder. 4055801 7 74 dep C9a 794:796 arg1 Mr 799:800 Mr 799:800 Mr = 28,000 799:809 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 7 75 theme carboxyl-terminal 860:876 arg1 segments 878:885 carboxyl-terminal segments 860:885 carboxyl-terminal segments 860:885 C9 can be cleaved by alpha-thrombin into two single-chain polypeptide fragments: C9a (Mr = 28,000) and C9b (Mr = 38,000), which are the amino- and carboxyl-terminal segments of the protein, respectively. 4055801 5 76 theme polypeptide 595:605 arg1 strands 584:590 radial strands 577:590 radial strands of polypeptide 577:605 Electron micrographs of poly(C9) indicate that the torus is formed by radial strands of polypeptide. 4055801 10 77 theme C9 1365:1366 arg1 cylinder 1369:1376 the poly(C9) cylinder 1356:1376 the poly(C9) cylinder 1356:1376 Determinants for the C9a region were observed about the torus, base, and on the midsection of the poly(C9) cylinder. 4055801 8 78 theme C9a 1154:1156 arg1 resistance 1083:1092 the resistance 1079:1092 the resistance of poly(C9) to denaturation 1079:1120 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 8 78 theme C9a 1154:1156 arg1 feature 1138:1144 a collective feature 1125:1144 a collective feature of both C9a and C9b 1125:1164 The cleaved form of the protein, C9a,b, can be induced to polymerize under suitable conditions to form sodium dodecyl sulfate-resistant poly(C9), indicating that the resistance of poly(C9) to denaturation is a collective feature of both C9a and C9b. 4055801 9 79 theme C9a 1171:1173 arg1 regions 1195:1201 The C9a and C9b polypeptide regions 1167:1201 The C9a and C9b polypeptide regions 1167:1201 The C9a and C9b polypeptide regions have been mapped on poly(C9) by immunoelectron microscopy. 4055801 1 80 theme poly 112:115 arg1 architecture 62:73 The architecture 58:73 The architecture of human complement component C9 and poly(C9) 58:119 The architecture of human complement component C9 and poly(C9) was investigated by transmission electron microscopy. 4055801 17 81 theme fold 2026:2029 arg1 parallel 2031:2038 at least one major fold parallel 2007:2038 at least one major fold parallel 2007:2038 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 2 82 theme = 199:199 arg1 Mr 196:197 Mr = 66,000 196:206 Mr = 66,000 196:206 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 2 82 theme = 199:199 arg1 C9 192:193 Monomeric native C9 175:193 Monomeric native C9 (Mr = 66,000) 175:207 Monomeric native C9 (Mr = 66,000) exhibits an ellipsoid appearance (70 X 50 A) with a crevice visible on one face. 4055801 17 83 theme central 2047:2053 arg1 axis 2055:2058 the central axis 2043:2058 the central axis of the tubule 2043:2072 To form a subunit of poly(C9) the polypeptide chain must form at least one major fold parallel to the central axis of the tubule. 4055801 15 84 theme single-domain 1875:1887 arg1 C9 1856:1857 C9 1856:1857 C9 1856:1857 In summary, C9 appears to be a single-domain protein. 4055801 15 84 theme single-domain 1875:1887 arg1 protein 1889:1895 a single-domain protein 1873:1895 a single-domain protein 1873:1895 In summary, C9 appears to be a single-domain protein. 24162852 3 0 theme Plasmodium-infected 694:712 arg1 erythrocytes 714:725 Plasmodium-infected erythrocytes 694:725 Plasmodium-infected erythrocytes 694:725 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 8 1 theme cavity 1402:1407 arg1 existence 1381:1389 the existence 1377:1389 the existence of a large cavity that traverses the entire length of the molecule 1377:1456 Remarkably, the crystal structure also shows the existence of a large cavity that traverses the entire length of the molecule. 24162852 9 2 theme SR-BI 1474:1478 arg1 Mutagenesis 1459:1469 Mutagenesis 1459:1469 Mutagenesis of SR-BI 1459:1478 Mutagenesis of SR-BI indicates that the cavity serves as a tunnel through which cholesterol(esters) are delivered from the bound lipoprotein to the outer leaflet of the plasma membrane. 24162852 10 3 theme selective 1831:1839 arg1 transfer 1847:1854 the selective lipid transfer 1827:1854 the selective lipid transfer characteristic of SR-BI and CD36 1827:1887 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 3 4 theme effete 677:682 arg1 cells 684:688 effete cells 677:688 effete cells 677:688 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 4 5 theme atherosclerosis 850:864 arg1 pathogenesis 834:845 the pathogenesis 830:845 the pathogenesis of atherosclerosis and of Alzheimer's disease 830:891 Notably, CD36 is also a receptor for modified lipoproteins and β-amyloid, and has been implicated in the pathogenesis of atherosclerosis and of Alzheimer's disease. 24162852 1 6 theme important 154:162 arg1 Members 87:93 Members 87:93 Members of the CD36 superfamily of scavenger receptor proteins 87:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 1 6 theme important 154:162 arg1 regulators 164:173 important regulators 154:173 important regulators of lipid metabolism and innate immunity 154:213 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 3 7 theme cells 684:688 arg1 phagocytosis 661:672 phagocytosis 661:672 phagocytosis of effete cells and Plasmodium-infected erythrocytes 661:725 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 3 8 theme lysosomal 500:508 arg1 delivery 510:517 lysosomal delivery 500:517 lysosomal delivery of β-glucocerebrosidase 500:541 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 2 9 theme molecular 296:304 arg1 patterns 306:313 pathogen-associated molecular patterns 276:313 pathogen-associated molecular patterns 276:313 They recognize normal and modified lipoproteins, as well as pathogen-associated molecular patterns. 24162852 10 10 theme ligands 1720:1726 arg1 constituents 1700:1711 lipidic constituents 1692:1711 lipidic constituents of the ligands attached to the receptor surface 1692:1759 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 10 11 theme SR-BI 1874:1878 arg1 characteristic 1856:1869 characteristic 1856:1869 characteristic 1856:1869 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 6 12 theme homology 1154:1161 arg1 modelling 1163:1171 homology modelling 1154:1171 homology modelling 1154:1171 Here we determine the crystal structure of LIMP-2 and infer, by homology modelling, the structure of SR-BI and CD36. 24162852 2 13 theme pathogen-associated 276:294 arg1 patterns 306:313 pathogen-associated molecular patterns 276:313 pathogen-associated molecular patterns 276:313 They recognize normal and modified lipoproteins, as well as pathogen-associated molecular patterns. 24162852 10 14 dep membrane 1783:1790 arg1 off 1772:1774 off 1772:1774 off 1772:1774 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 5 15 dep function 965:972 arg1 the 961:963 the 961:963 the 961:963 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 4 16 theme modified 766:773 arg1 lipoproteins 775:786 modified lipoproteins 766:786 modified lipoproteins 766:786 Notably, CD36 is also a receptor for modified lipoproteins and β-amyloid, and has been implicated in the pathogenesis of atherosclerosis and of Alzheimer's disease. 24162852 6 17 theme crystal 1112:1118 arg1 structure 1120:1128 the crystal structure 1108:1128 the crystal structure of LIMP-2 1108:1138 Here we determine the crystal structure of LIMP-2 and infer, by homology modelling, the structure of SR-BI and CD36. 24162852 3 18 theme erythrocytes 714:725 arg1 phagocytosis 661:672 phagocytosis 661:672 phagocytosis of effete cells and Plasmodium-infected erythrocytes 661:725 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 9 19 theme outer 1607:1611 arg1 leaflet 1613:1619 the outer leaflet 1603:1619 the outer leaflet of the plasma membrane 1603:1642 Mutagenesis of SR-BI indicates that the cavity serves as a tunnel through which cholesterol(esters) are delivered from the bound lipoprotein to the outer leaflet of the plasma membrane. 24162852 5 20 from roles 918:922 arg1 disease 938:944 disease 938:944 disease 938:944 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 5 20 from roles 918:922 arg1 health 927:932 health 927:932 health 927:932 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 3 21 theme β-glucocerebrosidase 522:541 arg1 delivery 510:517 lysosomal delivery 500:517 lysosomal delivery of β-glucocerebrosidase 500:541 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 10 22 theme lipidic 1692:1698 arg1 constituents 1700:1711 lipidic constituents 1692:1711 lipidic constituents of the ligands attached to the receptor surface 1692:1759 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 9 23 theme bound 1582:1586 arg1 lipoprotein 1588:1598 the bound lipoprotein 1578:1598 the bound lipoprotein 1578:1598 Mutagenesis of SR-BI indicates that the cavity serves as a tunnel through which cholesterol(esters) are delivered from the bound lipoprotein to the outer leaflet of the plasma membrane. 24162852 7 24 theme helical 1222:1228 arg1 bundle 1230:1235 a helical bundle 1220:1235 a helical bundle where β-glucocerebrosidase binds, and where ligands are most likely to bind to SR-BI and CD36 1220:1329 LIMP-2 shows a helical bundle where β-glucocerebrosidase binds, and where ligands are most likely to bind to SR-BI and CD36. 24162852 6 25 theme CD36 1201:1204 arg1 structure 1178:1186 the structure 1174:1186 the structure of SR-BI and CD36 1174:1204 Here we determine the crystal structure of LIMP-2 and infer, by homology modelling, the structure of SR-BI and CD36. 24162852 10 26 theme characteristic 1856:1869 arg1 transfer 1847:1854 the selective lipid transfer 1827:1854 the selective lipid transfer characteristic of SR-BI and CD36 1827:1887 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 1 27 theme lipid 178:182 arg1 metabolism 184:193 lipid metabolism 178:193 lipid metabolism 178:193 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 1 28 theme metabolism 184:193 arg1 Members 87:93 Members 87:93 Members of the CD36 superfamily of scavenger receptor proteins 87:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 1 28 theme metabolism 184:193 arg1 regulators 164:173 important regulators 154:173 important regulators of lipid metabolism and innate immunity 154:213 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 3 29 dep consists 327:334 arg1 LIMP-2/LGP85 471:482 LIMP-2/LGP85 471:482 LIMP-2/LGP85 471:482 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 3 29 dep consists 327:334 arg1 SR-BI 354:358 SR-BI 354:358 SR-BI 354:358 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 3 29 dep consists 327:334 arg1 CD36 613:616 CD36 613:616 CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes) 613:726 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 3 29 dep consists 327:334 arg1 receptor 559:566 a receptor 557:566 a receptor for enterovirus 71 and coxsackieviruses 557:606 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 6 30 theme LIMP-2 1133:1138 arg1 structure 1120:1128 the crystal structure 1108:1128 the crystal structure of LIMP-2 1108:1138 Here we determine the crystal structure of LIMP-2 and infer, by homology modelling, the structure of SR-BI and CD36. 24162852 8 31 theme crystal 1348:1354 arg1 structure 1356:1364 the crystal structure 1344:1364 the crystal structure 1344:1364 Remarkably, the crystal structure also shows the existence of a large cavity that traverses the entire length of the molecule. 24162852 3 32 theme C 461:461 arg1 virus 463:467 hepatitis C virus 451:467 hepatitis C virus 451:467 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 0 33 theme LIMP-2 13:18 arg1 Structure 0:8 Structure 0:8 Structure of LIMP-2 0:18 Structure of LIMP-2 provides functional insights with implications for SR-BI and CD36. 24162852 10 34 theme CD36 1884:1887 arg1 characteristic 1856:1869 characteristic 1856:1869 characteristic 1856:1869 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 9 35 theme plasma 1628:1633 arg1 membrane 1635:1642 the plasma membrane 1624:1642 the plasma membrane 1624:1642 Mutagenesis of SR-BI indicates that the cavity serves as a tunnel through which cholesterol(esters) are delivered from the bound lipoprotein to the outer leaflet of the plasma membrane. 24162852 8 36 theme large 1396:1400 arg1 cavity 1402:1407 a large cavity 1394:1407 a large cavity that traverses the entire length of the molecule 1394:1456 Remarkably, the crystal structure also shows the existence of a large cavity that traverses the entire length of the molecule. 24162852 1 37 theme innate 199:204 arg1 immunity 206:213 innate immunity 199:213 innate immunity 199:213 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 3 38 theme steroidogenic 405:417 arg1 organs 419:424 the liver and steroidogenic organs 391:424 the liver and steroidogenic organs 391:424 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 5 39 theme prominent 908:916 arg1 roles 918:922 their prominent roles 902:922 their prominent roles in health and disease 902:944 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 9 40 theme membrane 1635:1642 arg1 leaflet 1613:1619 the outer leaflet 1603:1619 the outer leaflet of the plasma membrane 1603:1642 Mutagenesis of SR-BI indicates that the cavity serves as a tunnel through which cholesterol(esters) are delivered from the bound lipoprotein to the outer leaflet of the plasma membrane. 24162852 3 41 dep CD36 613:616 arg1 transporter 632:642 a fatty-acid transporter 619:642 a fatty-acid transporter 619:642 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 3 41 dep CD36 613:616 arg1 receptor 648:655 receptor 648:655 receptor 648:655 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 5 42 theme family 1004:1009 arg1 members 1011:1017 the CD36 family members 995:1017 the CD36 family members 995:1017 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 1 43 theme immunity 206:213 arg1 Members 87:93 Members 87:93 Members of the CD36 superfamily of scavenger receptor proteins 87:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 1 43 theme immunity 206:213 arg1 regulators 164:173 important regulators 154:173 important regulators of lipid metabolism and innate immunity 154:213 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 0 44 theme functional 29:38 arg1 insights 40:47 functional insights 29:47 functional insights with implications for SR-BI and CD36 29:84 Structure of LIMP-2 provides functional insights with implications for SR-BI and CD36. 24162852 2 45 theme normal 231:236 arg1 lipoproteins 251:262 normal and modified lipoproteins 231:262 normal and modified lipoproteins 231:262 They recognize normal and modified lipoproteins, as well as pathogen-associated molecular patterns. 24162852 3 46 theme hepatitis 451:459 arg1 virus 463:467 hepatitis C virus 451:467 hepatitis C virus 451:467 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 3 47 theme liver 395:399 arg1 organs 419:424 the liver and steroidogenic organs 391:424 the liver and steroidogenic organs 391:424 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 9 48 theme cholesterol 1539:1549 arg1 esters 1551:1556 cholesterol(esters) 1539:1557 cholesterol(esters) 1539:1557 Mutagenesis of SR-BI indicates that the cavity serves as a tunnel through which cholesterol(esters) are delivered from the bound lipoprotein to the outer leaflet of the plasma membrane. 24162852 3 49 theme fatty-acid 621:630 arg1 transporter 632:642 a fatty-acid transporter 619:642 a fatty-acid transporter 619:642 The family consists of three members: SR-BI (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis C virus), LIMP-2/LGP85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 and coxsackieviruses) and CD36 (a fatty-acid transporter and receptor for phagocytosis of effete cells and Plasmodium-infected erythrocytes). 24162852 10 50 theme receptor 1744:1751 arg1 surface 1753:1759 the receptor surface 1740:1759 the receptor surface 1740:1759 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 1 51 theme CD36 102:105 arg1 superfamily 107:117 the CD36 superfamily 98:117 the CD36 superfamily of scavenger receptor proteins 98:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 8 52 theme molecule 1449:1456 arg1 length 1435:1440 the entire length 1424:1440 the entire length of the molecule 1424:1456 Remarkably, the crystal structure also shows the existence of a large cavity that traverses the entire length of the molecule. 24162852 6 53 theme SR-BI 1191:1195 arg1 structure 1178:1186 the structure 1174:1186 the structure of SR-BI and CD36 1174:1204 Here we determine the crystal structure of LIMP-2 and infer, by homology modelling, the structure of SR-BI and CD36. 24162852 1 54 theme superfamily 107:117 arg1 Members 87:93 Members 87:93 Members of the CD36 superfamily of scavenger receptor proteins 87:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 1 54 theme superfamily 107:117 arg1 regulators 164:173 important regulators 154:173 important regulators of lipid metabolism and innate immunity 154:213 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 5 55 theme CD36 999:1002 arg1 members 1011:1017 the CD36 family members 995:1017 the CD36 family members 995:1017 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 5 56 theme members 1011:1017 arg1 abnormalities 978:990 abnormalities 978:990 abnormalities 978:990 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 5 56 theme members 1011:1017 arg1 function 965:972 function 965:972 function 965:972 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 7 57 dep bundle 1230:1235 arg1 likely 1298:1303 likely 1298:1303 likely 1298:1303 LIMP-2 shows a helical bundle where β-glucocerebrosidase binds, and where ligands are most likely to bind to SR-BI and CD36. 24162852 7 57 dep bundle 1230:1235 arg1 binds 1264:1268 binds 1264:1268 binds 1264:1268 LIMP-2 shows a helical bundle where β-glucocerebrosidase binds, and where ligands are most likely to bind to SR-BI and CD36. 24162852 5 58 theme information 1055:1065 arg1 paucity 1044:1050 the paucity 1040:1050 the paucity of information about their structure 1040:1087 Despite their prominent roles in health and disease, understanding the function and abnormalities of the CD36 family members has been hampered by the paucity of information about their structure. 24162852 1 59 theme scavenger 122:130 arg1 proteins 141:148 scavenger receptor proteins 122:148 scavenger receptor proteins 122:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 8 60 theme entire 1428:1433 arg1 length 1435:1440 the entire length 1424:1440 the entire length of the molecule 1424:1456 Remarkably, the crystal structure also shows the existence of a large cavity that traverses the entire length of the molecule. 24162852 1 61 theme receptor 132:139 arg1 proteins 141:148 scavenger receptor proteins 122:148 scavenger receptor proteins 122:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 24162852 10 62 attach attached 1728:1735 arg2 ligands 1720:1726 the ligands 1716:1726 the ligands attached to the receptor surface 1716:1759 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 10 62 attach attached 1728:1735 arg1 surface 1753:1759 the receptor surface 1740:1759 the receptor surface 1740:1759 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 10 63 theme lipid 1841:1845 arg1 transfer 1847:1854 the selective lipid transfer 1827:1854 the selective lipid transfer characteristic of SR-BI and CD36 1827:1887 We provide evidence supporting a model whereby lipidic constituents of the ligands attached to the receptor surface are handed off to the membrane through the tunnel, accounting for the selective lipid transfer characteristic of SR-BI and CD36. 24162852 2 64 theme modified 242:249 arg1 lipoproteins 251:262 normal and modified lipoproteins 231:262 normal and modified lipoproteins 231:262 They recognize normal and modified lipoproteins, as well as pathogen-associated molecular patterns. 24162852 0 65 with insights 40:47 arg1 implications 54:65 implications 54:65 implications for SR-BI and CD36 54:84 Structure of LIMP-2 provides functional insights with implications for SR-BI and CD36. 24162852 1 66 theme proteins 141:148 arg1 superfamily 107:117 the CD36 superfamily 98:117 the CD36 superfamily of scavenger receptor proteins 98:148 Members of the CD36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. 6171497 1 0 from inhibitor 340:348 arg1 serum 359:363 human serum 353:363 human serum 353:363 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 0 1 theme inhibitor 92:100 arg1 proteolysis 53:63 limited proteolysis 45:63 limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography 45:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 2 theme affinity 186:193 arg1 chromatography 195:208 affinity chromatography 186:208 affinity chromatography 186:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 2 3 contain has 432:434 arg2 content 455:461 a 50% carbohydrate content 436:461 a 50% carbohydrate content 436:461 The inhibitor molecule has a 50% carbohydrate content. 6171497 2 3 contain has 432:434 arg1 molecule 423:430 The inhibitor molecule 409:430 The inhibitor molecule 409:430 The inhibitor molecule has a 50% carbohydrate content. 6171497 1 4 theme known 282:286 arg1 sequence 299:306 a known amino acid sequence 280:306 a known amino acid sequence 280:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 4 5 from residue 591:597 arg1 position 602:609 position 10 602:612 position 10 602:612 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 4 5 from residue 591:597 arg1 peptide 642:648 the N-terminal extension peptide 617:648 the N-terminal extension peptide 617:648 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 5 6 theme second 655:660 arg1 chain 667:671 The second side chain 651:671 The second side chain 651:671 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 7 from located 745:751 arg1 domain 794:799 the inactive "inhibitory" Kunitz-type domain 756:799 the inactive "inhibitory" Kunitz-type domain of the inhibitor 756:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 7 from located 745:751 arg1 inhibitor 808:816 the inhibitor 804:816 the inhibitor 804:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 0 8 from inhibitor 164:172 arg1 inhibitor 92:100 the inter-alpha-trypsin inhibitor 68:100 the inter-alpha-trypsin inhibitor 68:100 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 8 from inhibitor 164:172 arg1 Attachments 106:116 V. Attachments 103:116 V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography 103:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 9 theme V. 103:104 arg1 inhibitor 92:100 the inter-alpha-trypsin inhibitor 68:100 the inter-alpha-trypsin inhibitor 68:100 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 9 theme V. 103:104 arg1 Attachments 106:116 V. Attachments 103:116 V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography 103:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 5 10 theme side 662:665 arg1 chain 667:671 The second side chain 651:671 The second side chain 651:671 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 1 11 from bronchial 379:387 arg1 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 12 theme acid 294:297 arg1 sequence 299:306 a known amino acid sequence 280:306 a known amino acid sequence 280:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 0 13 theme carbohydrates 121:133 arg1 inhibitor 92:100 the inter-alpha-trypsin inhibitor 68:100 the inter-alpha-trypsin inhibitor 68:100 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 13 theme carbohydrates 121:133 arg1 Attachments 106:116 V. Attachments 103:116 V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography 103:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 1 14 from present 311:317 arg1 mucus 402:406 nasal mucus 396:406 nasal mucus 396:406 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 14 from present 311:317 arg1 urine 369:373 urine 369:373 urine 369:373 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 14 from present 311:317 arg1 bronchial 379:387 bronchial 379:387 bronchial 379:387 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 2 15 theme inhibitor 413:421 arg1 molecule 423:430 The inhibitor molecule 409:430 The inhibitor molecule 409:430 The inhibitor molecule has a 50% carbohydrate content. 6171497 4 16 theme serine 584:589 arg1 residue 591:597 the serine residue 580:597 the serine residue in position 10 in the N-terminal extension peptide 580:648 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 5 17 from domain 794:799 arg1 located 745:751 located 745:751 located 745:751 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 1 18 attach present 311:317 arg2 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 18 attach present 311:317 arg2 part 233:236 The inhibitory active part 211:236 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence 211:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 18 attach present 311:317 arg1 mucus 402:406 nasal mucus 396:406 nasal mucus 396:406 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 18 attach present 311:317 arg1 urine 369:373 urine 369:373 urine 369:373 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 18 attach present 311:317 arg1 bronchial 379:387 bronchial 379:387 bronchial 379:387 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 0 19 theme proteinase 12:21 arg1 inhibitors 23:32 Kunitz-type proteinase inhibitors 0:32 Kunitz-type proteinase inhibitors 0:32 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 5 20 theme inactive 760:767 arg1 domain 794:799 the inactive "inhibitory" Kunitz-type domain 756:799 the inactive "inhibitory" Kunitz-type domain of the inhibitor 756:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 20 theme inactive 760:767 arg1 inhibitor 808:816 the inhibitor 804:816 the inhibitor 804:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 21 theme asparagine 710:719 arg1 residue 721:727 the asparagine residue 706:727 the asparagine residue in position 24 706:742 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 0 22 theme Kunitz-type 0:10 arg1 inhibitors 23:32 Kunitz-type proteinase inhibitors 0:32 Kunitz-type proteinase inhibitors 0:32 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 1 23 from urine 369:373 arg1 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 5 24 from residue 721:727 arg1 position 732:739 position 24 732:742 position 24 732:742 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 1 25 theme inhibitory 215:224 arg1 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 25 theme inhibitory 215:224 arg1 part 233:236 The inhibitory active part 211:236 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence 211:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 5 26 theme inhibitor 808:816 arg1 domain 794:799 the inactive "inhibitory" Kunitz-type domain 756:799 the inactive "inhibitory" Kunitz-type domain of the inhibitor 756:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 26 theme inhibitor 808:816 arg1 inhibitor 808:816 the inhibitor 804:816 the inhibitor 804:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 2 27 theme 50 438:439 arg1 % 440:440 % 440:440 % 440:440 The inhibitor molecule has a 50% carbohydrate content. 6171497 6 28 theme chains 860:865 arg1 composition 823:833 The composition 819:833 The composition of the carbohydrate side chains 819:865 The composition of the carbohydrate side chains were determined. 6171497 4 29 attach linked 533:538 arg1 polypeptide 547:557 the polypeptide 543:557 the polypeptide 543:557 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 4 29 attach linked 533:538 arg3 residue 591:597 the serine residue 580:597 the serine residue in position 10 in the N-terminal extension peptide 580:648 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 4 29 attach linked 533:538 arg2 chain 524:528 One chain 520:528 One chain 520:528 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 1 30 theme active 226:231 arg1 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 30 theme active 226:231 arg1 part 233:236 The inhibitory active part 211:236 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence 211:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 0 31 theme limited 45:51 arg1 proteolysis 53:63 limited proteolysis 45:63 limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography 45:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 32 theme urinary 148:154 arg1 inhibitor 164:172 the human urinary trypsin inhibitor 138:172 the human urinary trypsin inhibitor isolated by affinity chromatography 138:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 6 33 theme side 855:858 arg1 chains 860:865 the carbohydrate side chains 838:865 the carbohydrate side chains 838:865 The composition of the carbohydrate side chains were determined. 6171497 5 34 theme inhibitory 770:779 arg1 domain 794:799 the inactive "inhibitory" Kunitz-type domain 756:799 the inactive "inhibitory" Kunitz-type domain of the inhibitor 756:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 34 theme inhibitory 770:779 arg1 inhibitor 808:816 the inhibitor 804:816 the inhibitor 804:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 35 dep attached 676:683 arg1 located 745:751 located 745:751 located 745:751 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 0 36 theme human 142:146 arg1 inhibitor 164:172 the human urinary trypsin inhibitor 138:172 the human urinary trypsin inhibitor isolated by affinity chromatography 138:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 1 37 theme nasal 396:400 arg1 mucus 402:406 nasal mucus 396:406 nasal mucus 396:406 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 3 38 attach attached 493:500 arg3 positions 509:517 two positions 505:517 two positions 505:517 Carbohydrate side chains are attached in two positions. 6171497 3 38 attach attached 493:500 arg2 chains 482:487 Carbohydrate side chains 464:487 Carbohydrate side chains 464:487 Carbohydrate side chains are attached in two positions. 6171497 4 39 theme extension 632:640 arg1 peptide 642:648 the N-terminal extension peptide 617:648 the N-terminal extension peptide 617:648 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 5 40 theme " 780:780 arg1 domain 794:799 the inactive "inhibitory" Kunitz-type domain 756:799 the inactive "inhibitory" Kunitz-type domain of the inhibitor 756:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 40 theme " 780:780 arg1 inhibitor 808:816 the inhibitor 804:816 the inhibitor 804:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 1 41 theme acid-resistant 325:338 arg1 inhibitor 340:348 an acid-resistant inhibitor 322:348 an acid-resistant inhibitor in human serum 322:363 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 5 42 theme Kunitz-type 782:792 arg1 domain 794:799 the inactive "inhibitory" Kunitz-type domain 756:799 the inactive "inhibitory" Kunitz-type domain of the inhibitor 756:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 42 theme Kunitz-type 782:792 arg1 inhibitor 808:816 the inhibitor 804:816 the inhibitor 804:816 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 5 43 dep located 745:751 arg1 N-glycosidically 685:700 N-glycosidically 685:700 N-glycosidically 685:700 The second side chain is attached N-glycosidically via the asparagine residue in position 24, located in the inactive "inhibitory" Kunitz-type domain of the inhibitor. 6171497 3 44 theme Carbohydrate 464:475 arg1 chains 482:487 Carbohydrate side chains 464:487 Carbohydrate side chains 464:487 Carbohydrate side chains are attached in two positions. 6171497 2 45 theme carbohydrate 442:453 arg1 content 455:461 a 50% carbohydrate content 436:461 a 50% carbohydrate content 436:461 The inhibitor molecule has a 50% carbohydrate content. 6171497 0 46 theme trypsin 156:162 arg1 inhibitor 164:172 the human urinary trypsin inhibitor 138:172 the human urinary trypsin inhibitor isolated by affinity chromatography 138:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 47 from carbohydrates 121:133 arg1 inhibitor 164:172 the human urinary trypsin inhibitor 138:172 the human urinary trypsin inhibitor isolated by affinity chromatography 138:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 6 48 theme carbohydrate 842:853 arg1 chains 860:865 the carbohydrate side chains 838:865 the carbohydrate side chains 838:865 The composition of the carbohydrate side chains were determined. 6171497 3 49 theme side 477:480 arg1 chains 482:487 Carbohydrate side chains 464:487 Carbohydrate side chains 464:487 Carbohydrate side chains are attached in two positions. 6171497 0 50 from Attachments 106:116 arg1 inhibitor 164:172 the human urinary trypsin inhibitor 138:172 the human urinary trypsin inhibitor isolated by affinity chromatography 138:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 1 51 with inhibitor 265:273 arg1 sequence 299:306 a known amino acid sequence 280:306 a known amino acid sequence 280:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 52 theme inter-alpha-trypsin 245:263 arg1 inhibitor 265:273 the inter-alpha-trypsin inhibitor 241:273 the inter-alpha-trypsin inhibitor with a known amino acid sequence 241:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 0 53 theme inter-alpha-trypsin 72:90 arg1 inhibitor 92:100 the inter-alpha-trypsin inhibitor 68:100 the inter-alpha-trypsin inhibitor 68:100 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 0 53 theme inter-alpha-trypsin 72:90 arg1 Attachments 106:116 V. Attachments 103:116 V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography 103:208 Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, V. Attachments of carbohydrates in the human urinary trypsin inhibitor isolated by affinity chromatography. 6171497 4 54 theme N-terminal 621:630 arg1 peptide 642:648 the N-terminal extension peptide 617:648 the N-terminal extension peptide 617:648 One chain is linked to the polypeptide O-glycosidically via the serine residue in position 10 in the N-terminal extension peptide. 6171497 1 55 theme inhibitor 265:273 arg1 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 55 theme inhibitor 265:273 arg1 part 233:236 The inhibitory active part 211:236 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence 211:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 56 theme amino 288:292 arg1 sequence 299:306 a known amino acid sequence 280:306 a known amino acid sequence 280:306 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 1 57 theme human 353:357 arg1 serum 359:363 human serum 353:363 human serum 353:363 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 6171497 2 58 theme % 440:440 arg1 content 455:461 a 50% carbohydrate content 436:461 a 50% carbohydrate content 436:461 The inhibitor molecule has a 50% carbohydrate content. 6171497 1 59 from mucus 402:406 arg1 present 311:317 present 311:317 present 311:317 The inhibitory active part of the inter-alpha-trypsin inhibitor with a known amino acid sequence is present as an acid-resistant inhibitor in human serum, in urine, in bronchial and in nasal mucus. 1556128 6 0 from differences 899:909 arg1 forms 926:930 the various forms 914:930 the various forms of HB-EGF 914:940 Some of the differences in the various forms of HB-EGF were found to be due to N-terminal heterogeneity. 1556128 3 1 theme cells 470:474 arg1 medium 454:459 the conditioned medium 438:459 the conditioned medium of U-937 cells 438:474 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 2 2 theme factor-like 162:172 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 2 theme factor-like 162:172 arg1 HB-EGF 189:194 HB-EGF 189:194 HB-EGF 189:194 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 2 theme factor-like 162:172 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 2 theme factor-like 162:172 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 3 theme family 262:267 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 3 theme family 262:267 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 3 theme family 262:267 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 9 4 theme conditioned 1422:1432 arg1 medium 1434:1439 U-937 cell conditioned medium 1411:1439 U-937 cell conditioned medium 1411:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 3 5 dep exchange 489:496 arg1 chromatography 577:590 reversed phase liquid chromatography 555:590 reversed phase liquid chromatography 555:590 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 8 6 theme electrophoresis 1233:1247 arg1 analysis 1249:1256 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis 1191:1256 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis 1191:1256 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 2 7 theme epidermal 145:153 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 7 theme epidermal 145:153 arg1 HB-EGF 189:194 HB-EGF 189:194 HB-EGF 189:194 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 7 theme epidermal 145:153 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 7 theme epidermal 145:153 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 8 theme EGF 257:259 arg1 family 262:267 the epidermal growth factor (EGF) family 228:267 the epidermal growth factor (EGF) family 228:267 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 4 9 theme first 620:624 arg1 round 626:630 the first round 616:630 the first round of C4 column chromatography 616:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 3 10 theme heparin 516:522 arg1 affinity 524:531 heparin affinity 516:531 heparin affinity 516:531 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 6 11 theme HB-EGF 935:940 arg1 forms 926:930 the various forms 914:930 the various forms of HB-EGF 914:940 Some of the differences in the various forms of HB-EGF were found to be due to N-terminal heterogeneity. 1556128 4 12 theme elution 597:603 arg1 profile 605:611 The elution profile 593:611 The elution profile of the first round of C4 column chromatography 593:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 7 13 theme mature 1048:1053 arg1 polypeptide 1062:1072 the mature HB-EGF polypeptide 1044:1072 the mature HB-EGF polypeptide 1044:1072 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 0 14 theme EGF-like 29:36 arg1 factor 45:50 heparin-binding EGF-like growth factor 13:50 heparin-binding EGF-like growth factor 13:50 Structure of heparin-binding EGF-like growth factor. 1556128 5 15 theme different 837:845 arg1 forms 858:862 different structural forms 837:862 different structural forms of the HB-EGF protein 837:884 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 8 16 theme sulfate-polyacrylamide 1206:1227 arg1 electrophoresis 1233:1247 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1191:1247 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis 1191:1256 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 8 17 contain have 1301:1304 arg2 masses 1325:1330 apparent molecular masses 1306:1330 apparent molecular masses of 19-23 kDa 1306:1343 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 8 17 contain have 1301:1304 arg1 forms 1285:1289 the various forms 1273:1289 the various forms of HB-EGF 1273:1299 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 8 18 theme HB-EGF 1294:1299 arg1 forms 1285:1289 the various forms 1273:1289 the various forms of HB-EGF 1273:1299 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 2 19 theme described 208:216 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 19 theme described 208:216 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 19 theme described 208:216 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 1 20 theme mature 113:118 arg1 protein 120:126 the mature protein 109:126 the mature protein 109:126 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 8 21 theme apparent 1306:1313 arg1 masses 1325:1330 apparent molecular masses 1306:1330 apparent molecular masses of 19-23 kDa 1306:1343 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 7 22 theme fragments 1019:1027 arg1 Microsequencing 992:1006 Microsequencing 992:1006 Microsequencing of tryptic fragments 992:1027 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 3 23 theme liquid 570:575 arg1 chromatography 577:590 reversed phase liquid chromatography 555:590 reversed phase liquid chromatography 555:590 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 4 24 theme growth 675:680 arg1 peaks 698:702 four growth factor activity peaks 670:702 four growth factor activity peaks with similar specific biological activities 670:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 3 25 theme reversed 555:562 arg1 chromatography 577:590 reversed phase liquid chromatography 555:590 reversed phase liquid chromatography 555:590 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 5 26 theme protein 878:884 arg1 forms 858:862 different structural forms 837:862 different structural forms of the HB-EGF protein 837:884 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 4 27 theme column 638:643 arg1 chromatography 645:658 C4 column chromatography 635:658 C4 column chromatography 635:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 2 28 theme factor 249:254 arg1 family 262:267 the epidermal growth factor (EGF) family 228:267 the epidermal growth factor (EGF) family 228:267 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 4 29 theme specific 717:724 arg1 activities 737:746 similar specific biological activities 709:746 similar specific biological activities 709:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 1 30 gly glycosylation 92:104 arg1 protein 120:126 the mature protein 109:126 the mature protein 109:126 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 5 31 contain contained 827:835 arg2 forms 858:862 different structural forms 837:862 different structural forms of the HB-EGF protein 837:884 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 5 31 contain contained 827:835 arg1 peaks 821:825 these peaks 815:825 these peaks 815:825 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 4 32 theme activity 689:696 arg1 peaks 698:702 four growth factor activity peaks 670:702 four growth factor activity peaks with similar specific biological activities 670:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 2 33 theme EGF 399:401 arg1 receptor 403:410 the EGF receptor 395:410 the EGF receptor 395:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 34 theme growth 242:247 arg1 family 262:267 the epidermal growth factor (EGF) family 228:267 the epidermal growth factor (EGF) family 228:267 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 0 35 theme heparin-binding 13:27 arg1 factor 45:50 heparin-binding EGF-like growth factor 13:50 heparin-binding EGF-like growth factor 13:50 Structure of heparin-binding EGF-like growth factor. 1556128 9 36 theme Further 1346:1352 arg1 analysis 1354:1361 Further analysis 1346:1361 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium 1346:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 0 37 theme growth 38:43 arg1 factor 45:50 heparin-binding EGF-like growth factor 13:50 heparin-binding EGF-like growth factor 13:50 Structure of heparin-binding EGF-like growth factor. 1556128 5 38 theme N-terminal 749:758 arg1 microsequencing 781:795 N-terminal and tryptic fragment microsequencing 749:795 N-terminal and tryptic fragment microsequencing 749:795 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 2 39 theme growth 174:179 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 39 theme growth 174:179 arg1 HB-EGF 189:194 HB-EGF 189:194 HB-EGF 189:194 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 39 theme growth 174:179 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 39 theme growth 174:179 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 5 40 theme tryptic 764:770 arg1 microsequencing 781:795 N-terminal and tryptic fragment microsequencing 749:795 N-terminal and tryptic fragment microsequencing 749:795 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 2 41 theme epidermal 232:240 arg1 family 262:267 the epidermal growth factor (EGF) family 228:267 the epidermal growth factor (EGF) family 228:267 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 9 42 theme form 1387:1390 arg1 analysis 1354:1361 Further analysis 1346:1361 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium 1346:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 4 43 theme round 626:630 arg1 profile 605:611 The elution profile 593:611 The elution profile of the first round of C4 column chromatography 593:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 2 44 theme 125I-EGF 333:340 arg1 binding 322:328 the binding 318:328 the binding of 125I-EGF to its receptor 318:356 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 9 45 theme HB-EGF 1395:1400 arg1 form 1387:1390 the most predominant form 1366:1390 the most predominant form of HB-EGF found in U-937 cell conditioned medium 1366:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 1 46 theme Multiple 53:60 arg1 forms 62:66 Multiple forms 53:66 Multiple forms 53:66 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 4 47 contain contained 660:668 arg1 profile 605:611 The elution profile 593:611 The elution profile of the first round of C4 column chromatography 593:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 4 47 contain contained 660:668 arg2 peaks 698:702 four growth factor activity peaks 670:702 four growth factor activity peaks with similar specific biological activities 670:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 3 48 theme U-937 464:468 arg1 cells 470:474 U-937 cells 464:474 U-937 cells 464:474 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 3 49 theme copper 499:504 arg1 affinity 506:513 copper affinity 499:513 copper affinity 499:513 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 2 50 theme growth 155:160 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 50 theme growth 155:160 arg1 HB-EGF 189:194 HB-EGF 189:194 HB-EGF 189:194 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 50 theme growth 155:160 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 50 theme growth 155:160 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 5 51 theme structural 847:856 arg1 forms 858:862 different structural forms 837:862 different structural forms of the HB-EGF protein 837:884 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 7 52 theme acids 1115:1119 arg1 acids 1115:1119 the 208 amino acids 1101:1119 the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule 1101:1188 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 7 52 theme acids 1115:1119 arg1 86 1095:1096 86 1095:1096 86 1095:1096 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 8 53 theme gel 1229:1231 arg1 electrophoresis 1233:1247 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1191:1247 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis 1191:1256 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 2 54 theme Heparin-binding 129:143 arg1 mitogenic 277:285 mitogenic 277:285 mitogenic 277:285 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 54 theme Heparin-binding 129:143 arg1 HB-EGF 189:194 HB-EGF 189:194 HB-EGF 189:194 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 54 theme Heparin-binding 129:143 arg1 factor 181:186 Heparin-binding epidermal growth factor-like growth factor 129:186 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) 129:195 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 54 theme Heparin-binding 129:143 arg1 member 218:223 a newly described member 200:223 a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor 200:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 2 55 theme BALB/c 291:296 arg1 cells 302:306 BALB/c 3T3 cells 291:306 BALB/c 3T3 cells 291:306 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 7 56 theme HB-EGF 1055:1060 arg1 polypeptide 1062:1072 the mature HB-EGF polypeptide 1044:1072 the mature HB-EGF polypeptide 1044:1072 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 8 57 theme various 1277:1283 arg1 forms 1285:1289 the various forms 1273:1289 the various forms of HB-EGF 1273:1299 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 4 58 with peaks 698:702 arg1 activities 737:746 similar specific biological activities 709:746 similar specific biological activities 709:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 9 59 theme 7.2-7.8 1471:1477 arg1 pI 1465:1466 a pI 1463:1466 a pI of 7.2-7.8 1463:1477 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 6 60 theme various 918:924 arg1 forms 926:930 the various forms 914:930 the various forms of HB-EGF 914:940 Some of the differences in the various forms of HB-EGF were found to be due to N-terminal heterogeneity. 1556128 4 61 theme chromatography 645:658 arg1 round 626:630 the first round 616:630 the first round of C4 column chromatography 616:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 7 62 contain contain 1078:1084 arg2 86 1095:1096 86 1095:1096 86 1095:1096 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 7 62 contain contain 1078:1084 arg2 acids 1115:1119 the 208 amino acids 1101:1119 the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule 1101:1188 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 7 62 contain contain 1078:1084 arg1 polypeptide 1062:1072 the mature HB-EGF polypeptide 1044:1072 the mature HB-EGF polypeptide 1044:1072 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 7 63 theme tryptic 1011:1017 arg1 fragments 1019:1027 tryptic fragments 1011:1027 tryptic fragments 1011:1027 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 3 64 theme phase 564:568 arg1 chromatography 577:590 reversed phase liquid chromatography 555:590 reversed phase liquid chromatography 555:590 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 1 65 theme protein 120:126 arg1 forms 62:66 Multiple forms 53:66 Multiple forms 53:66 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 1 65 theme protein 120:126 arg1 structure 77:85 primary structure 69:85 primary structure 69:85 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 1 65 theme protein 120:126 arg1 glycosylation 92:104 glycosylation 92:104 glycosylation of the mature protein 92:126 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 3 66 theme C4 552:553 arg1 rounds 542:547 two rounds 538:547 two rounds of C4 538:553 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 3 66 theme C4 552:553 arg1 exchange 489:496 cation exchange 482:496 cation exchange 482:496 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 3 66 theme C4 552:553 arg1 affinity 506:513 copper affinity 499:513 copper affinity 499:513 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 3 66 theme C4 552:553 arg1 affinity 524:531 heparin affinity 516:531 heparin affinity 516:531 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 5 67 theme HB-EGF 871:876 arg1 protein 878:884 the HB-EGF protein 867:884 the HB-EGF protein 867:884 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 8 68 theme molecular 1315:1323 arg1 masses 1325:1330 apparent molecular masses 1306:1330 apparent molecular masses of 19-23 kDa 1306:1343 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 6 69 theme N-terminal 966:975 arg1 heterogeneity 977:989 N-terminal heterogeneity 966:989 N-terminal heterogeneity 966:989 Some of the differences in the various forms of HB-EGF were found to be due to N-terminal heterogeneity. 1556128 9 70 gly O-glycosylated 1486:1499 arg1 it 1456:1457 it 1456:1457 it 1456:1457 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 4 71 theme C4 635:636 arg1 chromatography 645:658 C4 column chromatography 635:658 C4 column chromatography 635:658 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 7 72 theme precursor 1171:1179 arg1 molecule 1181:1188 the HB-EGF precursor molecule 1160:1188 the HB-EGF precursor molecule 1160:1188 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 8 73 theme kDa 1341:1343 arg1 masses 1325:1330 apparent molecular masses 1306:1330 apparent molecular masses of 19-23 kDa 1306:1343 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. 1556128 9 74 located found 1402:1406 arg2 form 1387:1390 the most predominant form 1366:1390 the most predominant form of HB-EGF found in U-937 cell conditioned medium 1366:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 9 74 located found 1402:1406 arg1 medium 1434:1439 U-937 cell conditioned medium 1411:1439 U-937 cell conditioned medium 1411:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 4 75 theme similar 709:715 arg1 activities 737:746 similar specific biological activities 709:746 similar specific biological activities 709:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 2 76 theme 3T3 298:300 arg1 cells 302:306 BALB/c 3T3 cells 291:306 BALB/c 3T3 cells 291:306 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 1 77 theme primary 69:75 arg1 structure 77:85 primary structure 69:85 primary structure 69:85 Multiple forms, primary structure, and glycosylation of the mature protein. 1556128 4 78 theme biological 726:735 arg1 activities 737:746 similar specific biological activities 709:746 similar specific biological activities 709:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 5 79 theme fragment 772:779 arg1 microsequencing 781:795 N-terminal and tryptic fragment microsequencing 749:795 N-terminal and tryptic fragment microsequencing 749:795 N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. 1556128 2 80 theme receptor 403:410 arg1 autophosphorylation 372:390 autophosphorylation 372:390 autophosphorylation of the EGF receptor 372:410 Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. 1556128 9 81 contain has 1459:1461 arg1 it 1456:1457 it 1456:1457 it 1456:1457 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 9 81 contain has 1459:1461 arg2 pI 1465:1466 a pI 1463:1466 a pI of 7.2-7.8 1463:1477 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 4 82 theme factor 682:687 arg1 peaks 698:702 four growth factor activity peaks 670:702 four growth factor activity peaks with similar specific biological activities 670:746 The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. 1556128 7 83 theme nucleotide 1134:1143 arg1 sequence 1145:1152 nucleotide sequence 1134:1152 nucleotide sequence 1134:1152 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 0 84 theme factor 45:50 arg1 Structure 0:8 Structure 0:8 Structure of heparin-binding EGF-like growth factor. 0:51 Structure of heparin-binding EGF-like growth factor. 1556128 7 85 theme HB-EGF 1164:1169 arg1 molecule 1181:1188 the HB-EGF precursor molecule 1160:1188 the HB-EGF precursor molecule 1160:1188 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 7 86 theme amino 1109:1113 arg1 acids 1115:1119 the 208 amino acids 1101:1119 the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule 1101:1188 Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. 1556128 9 87 theme predominant 1375:1385 arg1 form 1387:1390 the most predominant form 1366:1390 the most predominant form of HB-EGF found in U-937 cell conditioned medium 1366:1439 Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated. 1556128 3 88 theme conditioned 442:452 arg1 medium 454:459 the conditioned medium 438:459 the conditioned medium of U-937 cells 438:474 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 1556128 3 89 theme cation 482:487 arg1 exchange 489:496 cation exchange 482:496 cation exchange 482:496 HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. 26403559 2 0 theme cell 490:493 arg1 differentiation 495:509 haematopoietic stem cell differentiation 470:509 haematopoietic stem cell differentiation 470:509 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 1 1 theme disease 326:332 arg1 treatment 298:306 the treatment 294:306 the treatment of cardiovascular disease 294:332 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 2 theme active 159:164 arg1 domains 166:172 two active domains 155:172 two active domains 155:172 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 3 theme electrolyte 235:245 arg1 homeostasis 247:257 electrolyte homeostasis 235:257 electrolyte homeostasis 235:257 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 2 4 theme stem 485:488 arg1 differentiation 495:509 haematopoietic stem cell differentiation 470:509 haematopoietic stem cell differentiation 470:509 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 1 5 dep it 267:268 arg1 target 284:289 the principal target 270:289 it the principal target in the treatment of cardiovascular disease 267:332 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 5 6 theme domain 907:912 arg1 co-operation 914:925 domain co-operation 907:925 domain co-operation 907:925 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 1 7 theme homeostasis 247:257 arg1 regulation 202:211 the regulation 198:211 the regulation of blood pressure and electrolyte homeostasis 198:257 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 5 8 theme Ac-SDKP 895:901 arg1 study 886:890 A comprehensive kinetic study 862:890 A comprehensive kinetic study of Ac-SDKP and domain co-operation 862:925 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 4 9 theme Ac-SDKP 745:751 arg1 cleavage 753:760 Ac-SDKP cleavage 745:760 Ac-SDKP cleavage 745:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 4 10 with complex 706:712 arg1 products 733:740 the dipeptide products 719:740 the dipeptide products of Ac-SDKP cleavage 719:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 6 11 theme fibrosis 1208:1215 arg1 disorders 1217:1225 fibrosis disorders 1208:1225 fibrosis disorders 1208:1225 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 6 12 theme ACE 1132:1134 arg1 inhibitors 1136:1145 design novel ACE inhibitors 1119:1145 design novel ACE inhibitors 1119:1145 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 6 13 theme Ac-SDKP 1151:1157 arg1 analogues 1159:1167 Ac-SDKP analogues 1151:1167 Ac-SDKP analogues 1151:1167 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 6 14 theme N-domain 1082:1089 arg1 selectivity 1091:1101 N-domain selectivity 1082:1101 N-domain selectivity 1082:1101 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 1 15 theme Angiotensin-I 75:87 arg1 dipeptidylcarboxypeptidase 123:148 a zinc dipeptidylcarboxypeptidase 116:148 a zinc dipeptidylcarboxypeptidase 116:148 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 15 theme Angiotensin-I 75:87 arg1 ACE 108:110 ACE 108:110 ACE 108:110 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 15 theme Angiotensin-I 75:87 arg1 enzyme 100:105 Angiotensin-I converting enzyme 75:105 Angiotensin-I converting enzyme (ACE) 75:111 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 4 16 from structures 676:685 arg1 complex 706:712 complex 706:712 complex with the dipeptide products of Ac-SDKP cleavage 706:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 4 17 theme crystal 668:674 arg1 structures 676:685 The high resolution crystal structures 648:685 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage 648:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 2 18 theme negative 448:455 arg1 regulator 457:465 negative regulator 448:465 negative regulator of haematopoietic stem cell differentiation 448:509 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 1 19 from target 284:289 arg1 treatment 298:306 the treatment 294:306 the treatment of cardiovascular disease 294:332 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 20 theme converting 89:98 arg1 dipeptidylcarboxypeptidase 123:148 a zinc dipeptidylcarboxypeptidase 116:148 a zinc dipeptidylcarboxypeptidase 116:148 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 20 theme converting 89:98 arg1 ACE 108:110 ACE 108:110 ACE 108:110 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 20 theme converting 89:98 arg1 enzyme 100:105 Angiotensin-I converting enzyme 75:105 Angiotensin-I converting enzyme (ACE) 75:111 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 6 21 theme novel 1126:1130 arg1 inhibitors 1136:1145 design novel ACE inhibitors 1119:1145 design novel ACE inhibitors 1119:1145 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 4 22 theme resolution 657:666 arg1 structures 676:685 The high resolution crystal structures 648:685 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage 648:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 6 23 theme design 1119:1124 arg1 inhibitors 1136:1145 design novel ACE inhibitors 1119:1145 design novel ACE inhibitors 1119:1145 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 3 24 theme detailed 568:575 arg1 basis 604:608 a detailed biochemical and structural basis 566:608 a detailed biochemical and structural basis for the domain preference of Ac-SDKP 566:645 Here we provide a detailed biochemical and structural basis for the domain preference of Ac-SDKP. 26403559 5 25 theme kinetic 878:884 arg1 study 886:890 A comprehensive kinetic study 862:890 A comprehensive kinetic study of Ac-SDKP and domain co-operation 862:925 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 0 26 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 0:73 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 26403559 4 27 theme enzyme 854:859 arg1 recognition 835:845 substrate recognition 825:845 substrate recognition of the enzyme 825:859 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 3 28 theme biochemical 577:587 arg1 basis 604:608 a detailed biochemical and structural basis 566:608 a detailed biochemical and structural basis for the domain preference of Ac-SDKP 566:645 Here we provide a detailed biochemical and structural basis for the domain preference of Ac-SDKP. 26403559 1 29 theme key 186:188 arg1 role 190:193 a key role 184:193 a key role 184:193 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 0 30 theme Ac-SDKP 20:26 arg1 hydrolysis 28:37 Ac-SDKP hydrolysis 20:37 Ac-SDKP hydrolysis 20:37 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 26403559 3 31 theme structural 593:602 arg1 basis 604:608 a detailed biochemical and structural basis 566:608 a detailed biochemical and structural basis for the domain preference of Ac-SDKP 566:645 Here we provide a detailed biochemical and structural basis for the domain preference of Ac-SDKP. 26403559 2 32 theme potent 418:423 arg1 agent 438:442 a potent antifibrotic agent 416:442 a potent antifibrotic agent 416:442 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 1 33 theme principal 274:282 arg1 target 284:289 the principal target 270:289 it the principal target in the treatment of cardiovascular disease 267:332 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 5 34 theme domain 955:960 arg1 interactions 962:973 domain interactions 955:973 domain interactions affecting processing of the tetrapeptide substrate 955:1024 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 4 35 theme cleavage 753:760 arg1 products 733:740 the dipeptide products 719:740 the dipeptide products of Ac-SDKP cleavage 719:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 4 36 theme ACE 699:701 arg1 structures 676:685 The high resolution crystal structures 648:685 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage 648:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 2 37 theme haematopoietic 470:483 arg1 differentiation 495:509 haematopoietic stem cell differentiation 470:509 haematopoietic stem cell differentiation 470:509 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 6 38 used used 1183:1186 arg2 analogues 1159:1167 Ac-SDKP analogues 1151:1167 Ac-SDKP analogues 1151:1167 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 6 38 used used 1183:1186 arg2 inhibitors 1136:1145 design novel ACE inhibitors 1119:1145 design novel ACE inhibitors 1119:1145 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 4 39 theme high 652:655 arg1 structures 676:685 The high resolution crystal structures 648:685 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage 648:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 0 40 theme hydrolysis 28:37 arg1 basis 11:15 Structural basis 0:15 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 0:73 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 26403559 3 41 theme Ac-SDKP 639:645 arg1 preference 625:634 the domain preference 614:634 the domain preference of Ac-SDKP 614:645 Here we provide a detailed biochemical and structural basis for the domain preference of Ac-SDKP. 26403559 2 42 theme antifibrotic 425:436 arg1 agent 438:442 a potent antifibrotic agent 416:442 a potent antifibrotic agent 416:442 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 0 43 theme converting 56:65 arg1 enzyme 67:72 Angiotensin-I converting enzyme 42:72 Angiotensin-I converting enzyme 42:72 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 26403559 4 44 theme substrate 825:833 arg1 recognition 835:845 substrate recognition 825:845 substrate recognition of the enzyme 825:859 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 4 45 theme recognition 835:845 arg1 mechanism 812:820 the mechanism 808:820 the mechanism of substrate recognition of the enzyme 808:859 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 5 46 theme co-operation 914:925 arg1 study 886:890 A comprehensive kinetic study 862:890 A comprehensive kinetic study of Ac-SDKP and domain co-operation 862:925 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 0 47 theme Angiotensin-I 42:54 arg1 enzyme 67:72 Angiotensin-I converting enzyme 42:72 Angiotensin-I converting enzyme 42:72 Structural basis of Ac-SDKP hydrolysis by Angiotensin-I converting enzyme. 26403559 1 48 with dipeptidylcarboxypeptidase 123:148 arg1 domains 166:172 two active domains 155:172 two active domains 155:172 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 5 49 theme substrate 1016:1024 arg1 processing 985:994 processing 985:994 processing of the tetrapeptide substrate 985:1024 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 1 50 theme zinc 118:121 arg1 dipeptidylcarboxypeptidase 123:148 a zinc dipeptidylcarboxypeptidase 116:148 a zinc dipeptidylcarboxypeptidase 116:148 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 1 50 theme zinc 118:121 arg1 enzyme 100:105 Angiotensin-I converting enzyme 75:105 Angiotensin-I converting enzyme (ACE) 75:111 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 6 51 theme molecular 1062:1070 arg1 basis 1072:1076 the molecular basis 1058:1076 the molecular basis for N-domain selectivity 1058:1101 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 5 52 theme tetrapeptide 1003:1014 arg1 substrate 1016:1024 the tetrapeptide substrate 999:1024 the tetrapeptide substrate 999:1024 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 6 53 theme disorders 1217:1225 arg1 treatment 1195:1203 the treatment 1191:1203 the treatment of fibrosis disorders 1191:1225 Our results further illustrate the molecular basis for N-domain selectivity and should help design novel ACE inhibitors and Ac-SDKP analogues that could be used in the treatment of fibrosis disorders. 26403559 4 54 theme dipeptide 723:731 arg1 products 733:740 the dipeptide products 719:740 the dipeptide products of Ac-SDKP cleavage 719:760 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 26403559 1 55 theme blood 216:220 arg1 pressure 222:229 blood pressure 216:229 blood pressure 216:229 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 2 56 theme tetrapetide 354:364 arg1 Ac-SDKP 392:398 Ac-SDKP 392:398 Ac-SDKP 392:398 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 2 56 theme tetrapetide 354:364 arg1 N-acetyl-Ser-Asp-Lys-Pro 366:389 the tetrapetide N-acetyl-Ser-Asp-Lys-Pro 350:389 the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) 350:399 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 5 57 theme comprehensive 864:876 arg1 study 886:890 A comprehensive kinetic study 862:890 A comprehensive kinetic study of Ac-SDKP and domain co-operation 862:925 A comprehensive kinetic study of Ac-SDKP and domain co-operation was performed and indicated domain interactions affecting processing of the tetrapeptide substrate. 26403559 1 58 theme pressure 222:229 arg1 regulation 202:211 the regulation 198:211 the regulation of blood pressure and electrolyte homeostasis 198:257 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 2 59 theme differentiation 495:509 arg1 agent 438:442 a potent antifibrotic agent 416:442 a potent antifibrotic agent 416:442 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 2 59 theme differentiation 495:509 arg1 regulator 457:465 negative regulator 448:465 negative regulator of haematopoietic stem cell differentiation 448:509 More recently, the tetrapetide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) has emerged as a potent antifibrotic agent and negative regulator of haematopoietic stem cell differentiation which is processed exclusively by ACE. 26403559 1 60 theme cardiovascular 311:324 arg1 disease 326:332 cardiovascular disease 311:332 cardiovascular disease 311:332 Angiotensin-I converting enzyme (ACE) is a zinc dipeptidylcarboxypeptidase with two active domains and plays a key role in the regulation of blood pressure and electrolyte homeostasis, making it the principal target in the treatment of cardiovascular disease. 26403559 3 61 theme domain 618:623 arg1 preference 625:634 the domain preference 614:634 the domain preference of Ac-SDKP 614:645 Here we provide a detailed biochemical and structural basis for the domain preference of Ac-SDKP. 26403559 4 62 theme N-domain 690:697 arg1 ACE 699:701 N-domain ACE 690:701 N-domain ACE 690:701 The high resolution crystal structures of N-domain ACE in complex with the dipeptide products of Ac-SDKP cleavage were obtained and offered a template to model the mechanism of substrate recognition of the enzyme. 12200435 0 0 theme Sertoli 59:65 arg1 cells 67:71 Sertoli cells 59:71 Sertoli cells 59:71 The human "prion-like" protein Doppel is expressed in both Sertoli cells and spermatozoa. 12200435 1 1 theme protein 263:269 arg1 role 247:250 the physiological role 229:250 the physiological role of neither protein 229:269 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 2 2 theme Experimental 281:292 arg1 data 294:297 Experimental data 281:297 Experimental data 281:297 Experimental data suggest either direct or indirect interaction between the two proteins. 12200435 2 3 theme indirect 324:331 arg1 interaction 333:343 either direct or indirect interaction 307:343 either direct or indirect interaction between the two proteins 307:368 Experimental data suggest either direct or indirect interaction between the two proteins. 12200435 1 4 contain has 126:128 arg1 Dpl 121:123 Dpl 121:123 Dpl 121:123 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 1 4 contain has 126:128 arg1 protein 112:118 The prion-like Doppel protein 90:118 The prion-like Doppel protein (Dpl) 90:124 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 1 4 contain has 126:128 arg2 properties 162:171 many biochemical and structural properties 130:171 many biochemical and structural properties 130:171 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 4 5 link O-linked 670:677 arg1 sugars 679:684 O-linked sugars 670:684 O-linked sugars 670:684 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 3 6 theme biochemical 429:439 arg1 characteristics 441:455 biochemical characteristics 429:455 biochemical characteristics 429:455 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 4 7 theme O-linked 670:677 arg1 sugars 679:684 O-linked sugars 670:684 O-linked sugars 670:684 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 5 8 theme spermatozoa 773:783 arg1 flagella 739:746 the flagella 735:746 the flagella of epididymal and mature spermatozoa 735:783 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 4 9 with glycoprotein 645:656 arg1 N- 663:664 N- 663:664 N- 663:664 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 4 9 with glycoprotein 645:656 arg1 sugars 679:684 O-linked sugars 670:684 O-linked sugars 670:684 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 3 10 dep pattern 417:423 arg1 the 402:404 the 402:404 the 402:404 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 5 11 theme seminal 793:799 arg1 plasma 801:806 seminal plasma 793:806 seminal plasma 793:806 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 1 12 theme many 130:133 arg1 properties 162:171 many biochemical and structural properties 130:171 many biochemical and structural properties 130:171 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 3 13 theme human 467:471 arg1 tissues 473:479 human tissues 467:479 human tissues 467:479 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 1 14 theme biochemical 135:145 arg1 properties 162:171 many biochemical and structural properties 130:171 many biochemical and structural properties 130:171 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 0 15 theme human 4:8 arg1 Doppel 31:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel is expressed in both Sertoli cells and spermatozoa. 12200435 3 16 theme hamster 496:502 arg1 cells 510:514 Chinese hamster ovary cells 488:514 Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs 488:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 2 17 theme direct 314:319 arg1 interaction 333:343 either direct or indirect interaction 307:343 either direct or indirect interaction between the two proteins 307:368 Experimental data suggest either direct or indirect interaction between the two proteins. 12200435 7 18 theme human 1059:1063 arg1 fertility 1070:1078 human male fertility 1059:1078 human male fertility 1059:1078 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 19 theme germinal 982:989 arg1 cells 991:995 germinal cells 982:995 germinal cells 982:995 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 19 theme germinal 982:989 arg1 spermatozoa 969:979 spermatozoa 969:979 spermatozoa (germinal cells) 969:996 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 3 20 theme ovary 504:508 arg1 cells 510:514 Chinese hamster ovary cells 488:514 Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs 488:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 1 21 theme structural 151:160 arg1 properties 162:171 many biochemical and structural properties 130:171 many biochemical and structural properties 130:171 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 0 22 theme prion-like 11:20 arg1 Doppel 31:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel is expressed in both Sertoli cells and spermatozoa. 12200435 6 23 theme mature 877:882 arg1 spermatozoa 884:894 mature spermatozoa 877:894 mature spermatozoa 877:894 Dpl coexists only with N-terminally truncated isoforms of PrP(c) on mature spermatozoa. 12200435 1 24 with common 176:181 arg1 protein 207:213 cellular prion protein 192:213 the cellular prion protein (PrP(c)) 188:222 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 8 25 theme interesting 1137:1147 arg1 model 1149:1153 an interesting model 1134:1153 an interesting model for studies of the potential interaction between Dpl and PrP(c) 1134:1217 Finally, our data indicate that spermatozoa are thus an interesting model for studies of the potential interaction between Dpl and PrP(c). 12200435 8 25 theme interesting 1137:1147 arg1 spermatozoa 1113:1123 spermatozoa 1113:1123 spermatozoa 1113:1123 Finally, our data indicate that spermatozoa are thus an interesting model for studies of the potential interaction between Dpl and PrP(c). 12200435 5 26 theme epididymal 751:760 arg1 spermatozoa 773:783 epididymal and mature spermatozoa 751:783 epididymal and mature spermatozoa 751:783 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 3 27 theme Chinese 488:494 arg1 cells 510:514 Chinese hamster ovary cells 488:514 Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs 488:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 6 28 theme truncated 845:853 arg1 isoforms 855:862 N-terminally truncated isoforms 832:862 N-terminally truncated isoforms of PrP(c) 832:872 Dpl coexists only with N-terminally truncated isoforms of PrP(c) on mature spermatozoa. 12200435 5 29 theme mature 766:771 arg1 spermatozoa 773:783 epididymal and mature spermatozoa 751:783 epididymal and mature spermatozoa 751:783 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 0 30 theme protein 23:29 arg1 Doppel 31:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel is expressed in both Sertoli cells and spermatozoa. 12200435 1 31 dep protein 207:213 arg1 PrP 216:218 PrP 216:218 PrP(c) 216:221 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 1 31 dep protein 207:213 arg1 c 220:220 c 220:220 c 220:220 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 8 32 theme potential 1174:1182 arg1 interaction 1184:1194 the potential interaction 1170:1194 the potential interaction between Dpl and PrP(c) 1170:1217 Finally, our data indicate that spermatozoa are thus an interesting model for studies of the potential interaction between Dpl and PrP(c). 12200435 4 33 theme Human 581:585 arg1 glycoprotein 645:656 a glycosylphosphatidylinositol-anchored glycoprotein 605:656 a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars 605:684 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 4 33 theme Human 581:585 arg1 Dpl 587:589 Human Dpl 581:589 Human Dpl 581:589 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 0 34 theme " 21:21 arg1 Doppel 31:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel 0:36 The human "prion-like" protein Doppel is expressed in both Sertoli cells and spermatozoa. 12200435 3 35 theme gene 564:567 arg1 constructs 569:578 wild-type or variant human Dpl gene constructs 533:578 wild-type or variant human Dpl gene constructs 533:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 3 36 theme wild-type 533:541 arg1 constructs 569:578 wild-type or variant human Dpl gene constructs 533:578 wild-type or variant human Dpl gene constructs 533:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 1 37 theme physiological 233:245 arg1 role 247:250 the physiological role 229:250 the physiological role of neither protein 229:269 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 7 38 theme Sertoli 935:941 arg1 cells 943:947 Sertoli cells 935:947 Sertoli cells (somatic cells) 935:963 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 38 theme Sertoli 935:941 arg1 cells 958:962 somatic cells 950:962 somatic cells 950:962 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 8 39 theme interaction 1184:1194 arg1 studies 1159:1165 studies 1159:1165 studies of the potential interaction between Dpl and PrP(c) 1159:1217 Finally, our data indicate that spermatozoa are thus an interesting model for studies of the potential interaction between Dpl and PrP(c). 12200435 6 40 theme PrP 867:869 arg1 isoforms 855:862 N-terminally truncated isoforms 832:862 N-terminally truncated isoforms of PrP(c) 832:872 Dpl coexists only with N-terminally truncated isoforms of PrP(c) on mature spermatozoa. 12200435 7 41 theme somatic 950:956 arg1 cells 943:947 Sertoli cells 935:947 Sertoli cells (somatic cells) 935:963 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 41 theme somatic 950:956 arg1 cells 958:962 somatic cells 950:962 somatic cells 950:962 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 5 42 from cells 711:715 arg1 testis 724:729 the testis 720:729 the testis 720:729 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 7 43 theme human 917:921 arg1 Dpl 923:925 human Dpl 917:925 human Dpl 917:925 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 1 44 theme prion-like 94:103 arg1 Dpl 121:123 Dpl 121:123 Dpl 121:123 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 1 44 theme prion-like 94:103 arg1 protein 112:118 The prion-like Doppel protein 90:118 The prion-like Doppel protein (Dpl) 90:124 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 7 45 theme major 1045:1049 arg1 role 1051:1054 a major role 1043:1054 a major role 1043:1054 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 4 46 gly glycoprotein 645:656 arg1 glycoprotein 645:656 a glycosylphosphatidylinositol-anchored glycoprotein 605:656 a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars 605:684 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 4 46 gly glycoprotein 645:656 arg1 Dpl 587:589 Human Dpl 581:589 Human Dpl 581:589 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 3 47 theme human 554:558 arg1 constructs 569:578 wild-type or variant human Dpl gene constructs 533:578 wild-type or variant human Dpl gene constructs 533:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 7 48 theme Dpl 923:925 arg1 localization 901:912 The localization 897:912 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) 897:996 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 1 49 theme Doppel 105:110 arg1 Dpl 121:123 Dpl 121:123 Dpl 121:123 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 1 49 theme Doppel 105:110 arg1 protein 112:118 The prion-like Doppel protein 90:118 The prion-like Doppel protein (Dpl) 90:124 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 3 50 theme Dpl 560:562 arg1 constructs 569:578 wild-type or variant human Dpl gene constructs 533:578 wild-type or variant human Dpl gene constructs 533:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 3 51 theme expression 406:415 arg1 pattern 417:423 expression pattern 406:423 expression pattern 406:423 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 1 52 theme cellular 192:199 arg1 protein 207:213 cellular prion protein 192:213 the cellular prion protein (PrP(c)) 188:222 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 4 53 theme glycosylphosphatidylinositol-anchored 607:643 arg1 glycoprotein 645:656 a glycosylphosphatidylinositol-anchored glycoprotein 605:656 a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars 605:684 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 4 53 theme glycosylphosphatidylinositol-anchored 607:643 arg1 Dpl 587:589 Human Dpl 581:589 Human Dpl 581:589 Human Dpl appears to be a glycosylphosphatidylinositol-anchored glycoprotein with N- and O-linked sugars. 12200435 7 54 from localization 901:912 arg1 cells 991:995 germinal cells 982:995 germinal cells 982:995 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 54 from localization 901:912 arg1 cells 943:947 Sertoli cells 935:947 Sertoli cells (somatic cells) 935:963 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 54 from localization 901:912 arg1 cells 958:962 somatic cells 950:962 somatic cells 950:962 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 7 54 from localization 901:912 arg1 spermatozoa 969:979 spermatozoa 969:979 spermatozoa (germinal cells) 969:996 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 1 55 theme prion 201:205 arg1 protein 207:213 cellular prion protein 192:213 the cellular prion protein (PrP(c)) 188:222 The prion-like Doppel protein (Dpl) has many biochemical and structural properties in common with the cellular prion protein (PrP(c)), and the physiological role of neither protein is known. 12200435 5 56 theme Sertoli 703:709 arg1 cells 711:715 Sertoli cells 703:715 Sertoli cells in the testis 703:729 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 7 57 theme male 1065:1068 arg1 fertility 1070:1078 human male fertility 1059:1078 human male fertility 1059:1078 The localization of human Dpl on both Sertoli cells (somatic cells) and spermatozoa (germinal cells) strongly suggests that this protein may play a major role in human male fertility. 12200435 3 58 theme variant 546:552 arg1 constructs 569:578 wild-type or variant human Dpl gene constructs 533:578 wild-type or variant human Dpl gene constructs 533:578 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 5 59 located found 694:698 arg1 cells 711:715 Sertoli cells 703:715 Sertoli cells in the testis 703:729 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 5 59 located found 694:698 arg2 It 687:688 It 687:688 It 687:688 It was found on Sertoli cells in the testis, on the flagella of epididymal and mature spermatozoa, and in seminal plasma. 12200435 3 60 theme Dpl 460:462 arg1 pattern 417:423 expression pattern 406:423 expression pattern 406:423 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 12200435 3 60 theme Dpl 460:462 arg1 characteristics 441:455 biochemical characteristics 429:455 biochemical characteristics 429:455 In this study, we investigated the expression pattern and biochemical characteristics of Dpl in human tissues and in Chinese hamster ovary cells transfected with wild-type or variant human Dpl gene constructs. 18065761 3 0 theme ten 710:712 arg1 subunits 721:728 eight (dp8) and ten (dp10) subunits 694:728 eight (dp8) and ten (dp10) subunits 694:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 2 1 theme heparin-binding 371:385 arg1 TSP-1 397:401 TSP-1 397:401 TSP-1 397:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 2 1 theme heparin-binding 371:385 arg1 domain 387:392 the high affinity heparin-binding domain 353:392 the high affinity heparin-binding domain of TSP-1 353:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 5 2 theme heparin 1120:1126 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 3 from importance 1005:1014 arg1 positions 1040:1048 positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin 1040:1126 positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin 1040:1126 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 4 theme binding 1068:1074 arg1 groups 1084:1089 binding sulfate groups 1068:1089 binding sulfate groups of the dp8 and dp10 forms of heparin 1068:1126 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 4 theme binding 1068:1074 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 2 5 theme affinity 362:369 arg1 TSP-1 397:401 TSP-1 397:401 TSP-1 397:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 2 5 theme affinity 362:369 arg1 domain 387:392 the high affinity heparin-binding domain 353:392 the high affinity heparin-binding domain of TSP-1 353:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 1 6 theme matrix 265:270 arg1 structure 272:280 matrix structure 265:280 matrix structure 265:280 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 4 7 theme TSPN-1 895:900 arg1 dimers 902:907 trans and cis TSPN-1 dimers 881:907 trans and cis TSPN-1 dimers 881:907 We have found that dp8 and dp10 bind to TSPN-1 in a manner similar to Arixtra and that dp8 and dp10 induce the formation of trans and cis TSPN-1 dimers, respectively. 18065761 5 8 theme sulfate 1076:1082 arg1 groups 1084:1089 binding sulfate groups 1068:1089 binding sulfate groups of the dp8 and dp10 forms of heparin 1068:1126 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 8 theme sulfate 1076:1082 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 9 theme trans 1314:1318 arg1 dimers 1320:1325 cis and trans dimers 1306:1325 cis and trans dimers of the TSPN-1 domain 1306:1346 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 2 10 from structure 340:348 arg1 association 426:436 association 426:436 association with the synthetic heparin, Arixtra 426:472 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 8 11 theme TSPN-1 1524:1529 arg1 domains 1531:1537 TSPN-1 domains 1524:1537 TSPN-1 domains from two separate TSP-1 molecules 1524:1571 Dimer formation may also involve TSPN-1 domains from two separate TSP-1 molecules. 18065761 7 12 theme heparin 1382:1388 arg1 segments 1370:1377 relatively short segments 1353:1377 relatively short segments of heparin 1353:1388 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 4 13 theme cis 891:893 arg1 dimers 902:907 trans and cis TSPN-1 dimers 881:907 trans and cis TSPN-1 dimers 881:907 We have found that dp8 and dp10 bind to TSPN-1 in a manner similar to Arixtra and that dp8 and dp10 induce the formation of trans and cis TSPN-1 dimers, respectively. 18065761 3 14 theme association 551:561 arg1 binding 497:503 the binding 493:503 the binding of TSPN-1 to Arixtra 493:524 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 3 14 theme association 551:561 arg1 representative 529:542 representative 529:542 representative 529:542 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 3 15 theme TSPN-1 508:513 arg1 binding 497:503 the binding 493:503 the binding of TSPN-1 to Arixtra 493:524 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 3 15 theme TSPN-1 508:513 arg1 representative 529:542 representative 529:542 representative 529:542 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 5 16 dep In 924:925 arg1 silico 927:932 silico 927:932 silico 927:932 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 17 theme In 924:925 arg1 calculations 942:953 In silico docking calculations 924:953 In silico docking calculations partnered with our crystal structures 924:991 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 3 18 from structures 621:630 arg1 complex 645:651 complex 645:651 complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits 645:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 3 19 contain containing 683:692 arg2 subunits 721:728 eight (dp8) and ten (dp10) subunits 694:728 eight (dp8) and ten (dp10) subunits 694:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 3 19 contain containing 683:692 arg1 oligosaccharides 666:681 heparin oligosaccharides 658:681 heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits 658:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 4 20 theme dimers 902:907 arg1 formation 868:876 the formation 864:876 the formation of trans and cis TSPN-1 dimers 864:907 We have found that dp8 and dp10 bind to TSPN-1 in a manner similar to Arixtra and that dp8 and dp10 induce the formation of trans and cis TSPN-1 dimers, respectively. 18065761 8 21 theme Dimer 1491:1495 arg1 formation 1497:1505 Dimer formation 1491:1505 Dimer formation 1491:1505 Dimer formation may also involve TSPN-1 domains from two separate TSP-1 molecules. 18065761 8 22 theme separate 1548:1555 arg1 molecules 1563:1571 two separate TSP-1 molecules 1544:1571 two separate TSP-1 molecules 1544:1571 Dimer formation may also involve TSPN-1 domains from two separate TSP-1 molecules. 18065761 0 23 theme cis- 16:19 arg1 modes 44:48 Heparin-induced cis- and trans-dimerization modes 0:48 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 0:91 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 18065761 2 24 with association 426:436 arg1 heparin 457:463 the synthetic heparin 443:463 the synthetic heparin 443:463 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 2 24 with association 426:436 arg1 Arixtra 466:472 Arixtra 466:472 Arixtra 466:472 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 3 25 theme x-ray 734:738 arg1 crystallography 740:754 x-ray crystallography 734:754 x-ray crystallography 734:754 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 3 26 theme occurring 578:586 arg1 heparins 588:595 naturally occurring heparins 568:595 naturally occurring heparins 568:595 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 0 27 theme Heparin-induced 0:14 arg1 modes 44:48 Heparin-induced cis- and trans-dimerization modes 0:48 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 0:91 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 18065761 1 28 theme cellular 286:293 arg1 phenotype 295:303 cellular phenotype 286:303 cellular phenotype 286:303 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 4 29 theme similar 816:822 arg1 manner 809:814 a manner 807:814 a manner similar to Arixtra 807:833 We have found that dp8 and dp10 bind to TSPN-1 in a manner similar to Arixtra and that dp8 and dp10 induce the formation of trans and cis TSPN-1 dimers, respectively. 18065761 8 30 from molecules 1563:1571 arg1 domains 1531:1537 TSPN-1 domains 1524:1537 TSPN-1 domains from two separate TSP-1 molecules 1524:1571 Dimer formation may also involve TSPN-1 domains from two separate TSP-1 molecules. 18065761 2 31 theme TSP-1 397:401 arg1 TSP-1 397:401 TSP-1 397:401 TSP-1 397:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 2 31 theme TSP-1 397:401 arg1 domain 387:392 the high affinity heparin-binding domain 353:392 the high affinity heparin-binding domain of TSP-1 353:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 0 32 theme trans-dimerization 25:42 arg1 modes 44:48 Heparin-induced cis- and trans-dimerization modes 0:48 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 0:91 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 18065761 5 33 theme dp10 1106:1109 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 34 theme TSPN-1 1334:1339 arg1 domain 1341:1346 the TSPN-1 domain 1330:1346 the TSPN-1 domain 1330:1346 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 3 35 with complex 645:651 arg1 oligosaccharides 666:681 heparin oligosaccharides 658:681 heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits 658:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 1 36 with interactions 105:116 arg1 proteoglycans 136:148 proteoglycans 136:148 proteoglycans 136:148 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 1 36 with interactions 105:116 arg1 proteins 123:130 proteins 123:130 proteins 123:130 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 5 37 theme forms 1111:1115 arg1 groups 1084:1089 binding sulfate groups 1068:1089 binding sulfate groups of the dp8 and dp10 forms of heparin 1068:1126 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 37 theme forms 1111:1115 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 38 theme short 1364:1368 arg1 segments 1370:1377 relatively short segments 1353:1377 relatively short segments of heparin 1353:1388 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 5 39 theme residues 1028:1035 arg1 importance 1005:1014 the importance 1001:1014 the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin 1001:1126 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 40 theme domain 1341:1346 arg1 dimers 1320:1325 cis and trans dimers 1306:1325 cis and trans dimers of the TSPN-1 domain 1306:1346 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 1 41 theme cell 210:213 arg1 membrane 215:222 the cell membrane 206:222 the cell membrane 206:222 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 3 42 theme heparin 658:664 arg1 oligosaccharides 666:681 heparin oligosaccharides 658:681 heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits 658:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 6 43 theme domains 1159:1165 arg1 ability 1133:1139 The ability 1129:1139 The ability of several TSPN-1 domains to bind to glycosaminoglycans 1129:1195 The ability of several TSPN-1 domains to bind to glycosaminoglycans simultaneously probably increases the affinity of binding through multivalent interactions. 18065761 1 44 theme membrane 215:222 arg1 interface 193:201 the interface 189:201 the interface of the cell membrane 189:222 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 1 44 theme membrane 215:222 arg1 matrix 246:251 the extracellular matrix 228:251 the extracellular matrix 228:251 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 4 45 theme trans 881:885 arg1 dimers 902:907 trans and cis TSPN-1 dimers 881:907 trans and cis TSPN-1 dimers 881:907 We have found that dp8 and dp10 bind to TSPN-1 in a manner similar to Arixtra and that dp8 and dp10 induce the formation of trans and cis TSPN-1 dimers, respectively. 18065761 2 46 theme synthetic 447:455 arg1 heparin 457:463 the synthetic heparin 443:463 the synthetic heparin 443:463 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 2 46 theme synthetic 447:455 arg1 Arixtra 466:472 Arixtra 466:472 Arixtra 466:472 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 0 47 theme thrombospondin-1 57:72 arg1 domain 85:90 the thrombospondin-1 N-terminal domain 53:90 the thrombospondin-1 N-terminal domain 53:90 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 18065761 5 48 from positions 1040:1048 arg1 groups 1084:1089 binding sulfate groups 1068:1089 binding sulfate groups of the dp8 and dp10 forms of heparin 1068:1126 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 48 from positions 1040:1048 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 6 49 theme several 1144:1150 arg1 domains 1159:1165 several TSPN-1 domains 1144:1165 several TSPN-1 domains 1144:1165 The ability of several TSPN-1 domains to bind to glycosaminoglycans simultaneously probably increases the affinity of binding through multivalent interactions. 18065761 2 50 theme high 357:360 arg1 TSP-1 397:401 TSP-1 397:401 TSP-1 397:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 2 50 theme high 357:360 arg1 domain 387:392 the high affinity heparin-binding domain 353:392 the high affinity heparin-binding domain of TSP-1 353:401 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 3 51 with association 551:561 arg1 heparins 588:595 naturally occurring heparins 568:595 naturally occurring heparins 568:595 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 6 52 theme TSPN-1 1152:1157 arg1 domains 1159:1165 several TSPN-1 domains 1144:1165 several TSPN-1 domains 1144:1165 The ability of several TSPN-1 domains to bind to glycosaminoglycans simultaneously probably increases the affinity of binding through multivalent interactions. 18065761 6 53 theme binding 1247:1253 arg1 affinity 1235:1242 the affinity 1231:1242 the affinity of binding 1231:1253 The ability of several TSPN-1 domains to bind to glycosaminoglycans simultaneously probably increases the affinity of binding through multivalent interactions. 18065761 3 54 theme eight 694:698 arg1 subunits 721:728 eight (dp8) and ten (dp10) subunits 694:728 eight (dp8) and ten (dp10) subunits 694:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 7 55 theme dimers 1320:1325 arg1 formation 1293:1301 The formation 1289:1301 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin 1289:1388 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 8 56 theme TSP-1 1557:1561 arg1 molecules 1563:1571 two separate TSP-1 molecules 1544:1571 two separate TSP-1 molecules 1544:1571 Dimer formation may also involve TSPN-1 domains from two separate TSP-1 molecules. 18065761 5 57 theme dp8 1098:1100 arg1 forms 1111:1115 the dp8 and dp10 forms 1094:1115 forms 1111:1115 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 58 theme cis 1306:1308 arg1 dimers 1320:1325 cis and trans dimers 1306:1325 cis and trans dimers of the TSPN-1 domain 1306:1346 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 5 59 theme docking 934:940 arg1 calculations 942:953 In silico docking calculations 924:953 In silico docking calculations partnered with our crystal structures 924:991 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 60 theme crystal 974:980 arg1 structures 982:991 our crystal structures 970:991 our crystal structures 970:991 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 61 theme TSP-1 1422:1426 arg1 ability 1411:1417 the ability 1407:1417 the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans 1407:1488 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 6 62 theme multivalent 1263:1273 arg1 interactions 1275:1286 multivalent interactions 1263:1286 multivalent interactions 1263:1286 The ability of several TSPN-1 domains to bind to glycosaminoglycans simultaneously probably increases the affinity of binding through multivalent interactions. 18065761 7 63 with formation 1293:1301 arg1 segments 1370:1377 relatively short segments 1353:1377 relatively short segments of heparin 1353:1388 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18065761 3 64 theme TSPN-1 635:640 arg1 structures 621:630 the structures 617:630 the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits 617:728 To establish that the binding of TSPN-1 to Arixtra is representative of the association with naturally occurring heparins, we have determined the structures of TSPN-1 in complex with heparin oligosaccharides containing eight (dp8) and ten (dp10) subunits, by x-ray crystallography. 18065761 1 65 theme extracellular 232:244 arg1 matrix 246:251 the extracellular matrix 228:251 the extracellular matrix 228:251 Through its interactions with proteins and proteoglycans, thrombospondin-1 (TSP-1) functions at the interface of the cell membrane and the extracellular matrix to regulate matrix structure and cellular phenotype. 18065761 0 66 theme domain 85:90 arg1 modes 44:48 Heparin-induced cis- and trans-dimerization modes 0:48 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 0:91 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 18065761 5 67 theme arginine 1019:1026 arg1 residues 1028:1035 arginine residues 1019:1035 arginine residues 1019:1035 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 5 68 theme partnered 955:963 arg1 calculations 942:953 In silico docking calculations 924:953 In silico docking calculations partnered with our crystal structures 924:991 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 0 69 theme N-terminal 74:83 arg1 domain 85:90 the thrombospondin-1 N-terminal domain 53:90 the thrombospondin-1 N-terminal domain 53:90 Heparin-induced cis- and trans-dimerization modes of the thrombospondin-1 N-terminal domain. 18065761 2 70 theme domain 387:392 arg1 structure 340:348 the structure 336:348 the structure 336:348 We have previously determined the structure of the high affinity heparin-binding domain of TSP-1, designated TSPN-1, in association with the synthetic heparin, Arixtra. 18065761 5 71 with partnered 955:963 arg1 structures 982:991 our crystal structures 970:991 our crystal structures 970:991 In silico docking calculations partnered with our crystal structures support the importance of arginine residues in positions 29, 42, and 77 in binding sulfate groups of the dp8 and dp10 forms of heparin. 18065761 7 72 theme high 1446:1449 arg1 affinity 1451:1458 high affinity 1446:1458 high affinity binding to glycosaminoglycans 1446:1488 The formation of cis and trans dimers of the TSPN-1 domain with relatively short segments of heparin further enhances the ability of TSP-1 to participate in high affinity binding to glycosaminoglycans. 18794910 4 0 located detected 695:702 arg1 lesions 720:726 precancerous lesions 707:726 precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 707:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 0 located detected 695:702 arg2 RNF13 685:689 RNF13 685:689 RNF13 685:689 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 0 located detected 695:702 arg1 addition 675:682 addition 675:682 addition 675:682 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 1 theme 23/29 833:837 arg1 % 847:847 23/29 and 62.8% 833:847 % 847:847 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 1 theme 23/29 833:837 arg1 % 830:830 79.3% 826:830 79.3% 826:830 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 1 2 theme cancer 153:158 arg1 development 160:170 cancer development 153:170 cancer development 153:170 Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. 18794910 4 3 theme 62.8 843:846 arg1 % 847:847 23/29 and 62.8% 833:847 % 847:847 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 3 theme 62.8 843:846 arg1 % 830:830 79.3% 826:830 79.3% 826:830 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 3 4 from over-expression 533:547 arg1 samples 561:567 30 tumor samples 552:567 30 tumor samples 552:567 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 2 5 theme E3 300:301 arg1 ligase 313:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 5 theme E3 300:301 arg1 RNF13 259:263 a RING-finger-containing protein RNF13 226:263 a RING-finger-containing protein RNF13 226:263 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 6 theme ubiquitin 303:311 arg1 ligase 313:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 6 theme ubiquitin 303:311 arg1 RNF13 259:263 a RING-finger-containing protein RNF13 226:263 a RING-finger-containing protein RNF13 226:263 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 6 7 theme domain-mutant 1084:1096 arg1 RNF13 1098:1102 RING domain-mutant RNF13 1079:1102 wild type but not RING domain-mutant RNF13 1061:1102 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 6 8 theme pancreatic 1107:1116 arg1 cells 1135:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 2 9 theme membrane-associated 280:298 arg1 ligase 313:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 9 theme membrane-associated 280:298 arg1 RNF13 259:263 a RING-finger-containing protein RNF13 226:263 a RING-finger-containing protein RNF13 226:263 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 6 10 theme RNF13 1098:1102 arg1 Over-expression 1042:1056 Over-expression 1042:1056 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells 1042:1139 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 4 11 dep lesions 720:726 arg1 complexes 737:745 tubular complexes 729:745 precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 707:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 5 12 theme cancer 1022:1027 arg1 progression 1029:1039 cancer progression 1022:1039 cancer progression 1022:1039 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 7 13 theme ubiquitin-mediated 1347:1364 arg1 modification 1366:1377 ubiquitin-mediated modification 1347:1377 ubiquitin-mediated modification of proteins by RNF13 1347:1398 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 3 14 theme tissues 491:497 arg1 analysis 414:421 Immunohistochemical analysis 394:421 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients 394:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 2 15 theme ER/Golgi 271:278 arg1 ligase 313:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 15 theme ER/Golgi 271:278 arg1 RNF13 259:263 a RING-finger-containing protein RNF13 226:263 a RING-finger-containing protein RNF13 226:263 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 7 16 theme ubiquitin 1291:1299 arg1 RNF13 1271:1275 RNF13 1271:1275 RNF13 1271:1275 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 7 16 theme ubiquitin 1291:1299 arg1 ligase 1301:1306 a novel E3 ubiquitin ligase 1280:1306 a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis 1280:1344 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 7 17 theme novel 1282:1286 arg1 RNF13 1271:1275 RNF13 1271:1275 RNF13 1271:1275 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 7 17 theme novel 1282:1286 arg1 ligase 1301:1306 a novel E3 ubiquitin ligase 1280:1306 a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis 1280:1344 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 5 18 from role 1005:1008 arg1 progression 1029:1039 cancer progression 1022:1039 cancer progression 1022:1039 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 7 19 theme pancreatic 1419:1428 arg1 development 1437:1447 pancreatic cancer development 1419:1447 pancreatic cancer development 1419:1447 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 3 20 theme pancreatic 426:435 arg1 PDAC 460:463 PDAC 460:463 PDAC 460:463 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 20 theme pancreatic 426:435 arg1 adenocarcinoma 444:457 pancreatic ductal adenocarcinoma 426:457 pancreatic ductal adenocarcinoma (PDAC) 426:464 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 4 21 from complexes 737:745 arg1 neoplasia 807:815 pancreatic intraepithelial neoplasia 780:815 pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 780:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 21 from complexes 737:745 arg1 CP 772:773 CP 772:773 CP 772:773 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 21 from complexes 737:745 arg1 pancreatitis 758:769 chronic pancreatitis 750:769 chronic pancreatitis (CP) 750:774 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 21 from complexes 737:745 arg1 PanIN 818:822 PanIN 818:822 PanIN 818:822 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 3 22 dep documented 516:525 arg1 % 574:574 41.7% 570:574 41.7% 570:574 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 5 23 from expression 942:951 arg1 samples 973:979 PDAC samples 968:979 PDAC samples 968:979 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 7 24 theme E3 1288:1289 arg1 RNF13 1271:1275 RNF13 1271:1275 RNF13 1271:1275 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 7 24 theme E3 1288:1289 arg1 ligase 1301:1306 a novel E3 ubiquitin ligase 1280:1306 a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis 1280:1344 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 7 25 theme cancer 1430:1435 arg1 development 1437:1447 pancreatic cancer development 1419:1447 pancreatic cancer development 1419:1447 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 6 26 theme matrix 1200:1205 arg1 metalloproteinase-9 1207:1225 matrix metalloproteinase-9 1200:1225 matrix metalloproteinase-9 1200:1225 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 5 27 theme P 954:954 arg1 expression 942:951 Tenascin-C expression 931:951 Tenascin-C expression (P = 0.004) in PDAC samples 931:979 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 5 27 theme P 954:954 arg1 0.004 958:962 P = 0.004 954:962 P = 0.004 954:962 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 1 28 theme Protein 79:85 arg1 ubiquitination 87:100 Protein ubiquitination 79:100 Protein ubiquitination by E3 ubiquitin ligases 79:124 Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. 18794910 3 29 theme adenocarcinoma 444:457 arg1 analysis 414:421 Immunohistochemical analysis 394:421 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients 394:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 30 theme Immunohistochemical 394:412 arg1 analysis 414:421 Immunohistochemical analysis 394:421 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients 394:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 31 from patients 507:514 arg1 PDAC 460:463 PDAC 460:463 PDAC 460:463 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 31 from patients 507:514 arg1 adenocarcinoma 444:457 pancreatic ductal adenocarcinoma 426:457 pancreatic ductal adenocarcinoma (PDAC) 426:464 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 31 from patients 507:514 arg1 analysis 414:421 Immunohistochemical analysis 394:421 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients 394:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 31 from patients 507:514 arg1 tissues 491:497 paracancerous normal tissues 470:497 paracancerous normal tissues from 72 patients 470:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 5 32 theme RNF13 882:886 arg1 staining 888:895 RNF13 staining 882:895 RNF13 staining 882:895 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 2 33 theme ligase 377:382 arg1 activity 384:391 the ubiquitin ligase activity 363:391 the ubiquitin ligase activity 363:391 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 0 34 theme novel 9:13 arg1 ligase 35:40 a novel RING-type ubiquitin ligase 7:40 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 0:77 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 18794910 4 35 theme pancreatic 780:789 arg1 neoplasia 807:815 pancreatic intraepithelial neoplasia 780:815 pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 780:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 35 theme pancreatic 780:789 arg1 PanIN 818:822 PanIN 818:822 PanIN 818:822 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 5 36 theme PDAC 968:971 arg1 samples 973:979 PDAC samples 968:979 PDAC samples 968:979 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 3 37 theme normal 484:489 arg1 tissues 491:497 paracancerous normal tissues 470:497 paracancerous normal tissues from 72 patients 470:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 6 38 theme RING 1079:1082 arg1 RNF13 1098:1102 RING domain-mutant RNF13 1079:1102 wild type but not RING domain-mutant RNF13 1061:1102 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 3 39 theme tumor 555:559 arg1 samples 561:567 30 tumor samples 552:567 30 tumor samples 552:567 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 6 40 theme MiaPaca-2 1118:1126 arg1 cells 1135:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 1 41 theme E3 105:106 arg1 ligases 118:124 E3 ubiquitin ligases 105:124 E3 ubiquitin ligases 105:124 Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. 18794910 1 42 theme important 135:143 arg1 role 145:148 an important role 132:148 an important role 132:148 Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. 18794910 0 43 theme ubiquitin 25:33 arg1 ligase 35:40 a novel RING-type ubiquitin ligase 7:40 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 0:77 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 18794910 5 44 theme = 956:956 arg1 expression 942:951 Tenascin-C expression 931:951 Tenascin-C expression (P = 0.004) in PDAC samples 931:979 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 5 44 theme = 956:956 arg1 0.004 958:962 P = 0.004 954:962 P = 0.004 954:962 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 3 45 theme P 660:660 arg1 0.024 664:668 P = 0.024 660:668 P = 0.024 660:668 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 45 theme P 660:660 arg1 grading 651:657 histological grading 638:657 histological grading (P = 0.024) 638:669 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 6 46 theme cancer 1128:1133 arg1 cells 1135:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 4 47 dep neoplasia 807:815 arg1 22/35 850:854 22/35 850:854 22/35 850:854 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 47 dep neoplasia 807:815 arg1 % 847:847 23/29 and 62.8% 833:847 % 847:847 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 47 dep neoplasia 807:815 arg1 % 830:830 79.3% 826:830 79.3% 826:830 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 0 48 theme RING-type 15:23 arg1 ligase 35:40 a novel RING-type ubiquitin ligase 7:40 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 0:77 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 18794910 3 49 theme ductal 437:442 arg1 PDAC 460:463 PDAC 460:463 PDAC 460:463 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 49 theme ductal 437:442 arg1 adenocarcinoma 444:457 pancreatic ductal adenocarcinoma 426:457 pancreatic ductal adenocarcinoma (PDAC) 426:464 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 50 theme histological 638:649 arg1 0.024 664:668 P = 0.024 660:668 P = 0.024 660:668 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 50 theme histological 638:649 arg1 grading 651:657 histological grading 638:657 histological grading (P = 0.024) 638:669 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 4 51 theme intraepithelial 791:805 arg1 neoplasia 807:815 pancreatic intraepithelial neoplasia 780:815 pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 780:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 51 theme intraepithelial 791:805 arg1 PanIN 818:822 PanIN 818:822 PanIN 818:822 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 3 52 theme paracancerous 470:482 arg1 tissues 491:497 paracancerous normal tissues 470:497 paracancerous normal tissues from 72 patients 470:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 5 53 theme RNF13 1013:1017 arg1 role 1005:1008 the role 1001:1008 the role of RNF13 in cancer progression 1001:1039 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 7 54 theme pancreatic 1320:1329 arg1 carcinogenesis 1331:1344 pancreatic carcinogenesis 1320:1344 pancreatic carcinogenesis 1320:1344 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 6 55 from Over-expression 1042:1056 arg1 cells 1135:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 pancreatic MiaPaca-2 cancer cells 1107:1139 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 1 56 theme ubiquitin 108:116 arg1 ligases 118:124 E3 ubiquitin ligases 105:124 E3 ubiquitin ligases 105:124 Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. 18794910 7 57 theme proteins 1382:1389 arg1 modification 1366:1377 ubiquitin-mediated modification 1347:1377 ubiquitin-mediated modification of proteins by RNF13 1347:1398 Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiquitin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development. 18794910 2 58 theme experimental 199:210 arg1 evidence 212:219 experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity 199:391 experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity 199:391 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 6 59 theme wild 1061:1064 arg1 type 1066:1069 wild type 1061:1069 wild type but not RING domain-mutant RNF13 1061:1102 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 0 60 dep RNF13 0:4 arg1 ligase 35:40 a novel RING-type ubiquitin ligase 7:40 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 0:77 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 18794910 6 61 theme invasive 1151:1158 arg1 potential 1160:1168 invasive potential 1151:1168 invasive potential 1151:1168 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 6 62 theme gelatinolytic 1174:1186 arg1 activity 1188:1195 gelatinolytic activity 1174:1195 gelatinolytic activity 1174:1195 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 2 63 theme finger 333:338 arg1 domain 340:345 its RING finger domain 324:345 its RING finger domain 324:345 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 5 64 theme Tenascin-C 931:940 arg1 expression 942:951 Tenascin-C expression 931:951 Tenascin-C expression (P = 0.004) in PDAC samples 931:979 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 5 64 theme Tenascin-C 931:940 arg1 0.004 958:962 P = 0.004 954:962 P = 0.004 954:962 Moreover, RNF13 staining was significantly correlated with Tenascin-C expression (P = 0.004) in PDAC samples, further supporting the role of RNF13 in cancer progression. 18794910 3 65 theme = 662:662 arg1 0.024 664:668 P = 0.024 660:668 P = 0.024 660:668 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 3 65 theme = 662:662 arg1 grading 651:657 histological grading 638:657 histological grading (P = 0.024) 638:669 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 4 66 theme chronic 750:756 arg1 CP 772:773 CP 772:773 CP 772:773 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 66 theme chronic 750:756 arg1 pancreatitis 758:769 chronic pancreatitis 750:769 chronic pancreatitis (CP) 750:774 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 4 67 theme tubular 729:735 arg1 complexes 737:745 tubular complexes 729:745 precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 707:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 2 68 theme RING 328:331 arg1 domain 340:345 its RING finger domain 324:345 its RING finger domain 324:345 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 6 69 theme type 1066:1069 arg1 Over-expression 1042:1056 Over-expression 1042:1056 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells 1042:1139 Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. 18794910 4 70 theme precancerous 707:718 arg1 lesions 720:726 precancerous lesions 707:726 precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively) 707:869 In addition, RNF13 was detected in precancerous lesions: tubular complexes in chronic pancreatitis (CP) and pancreatic intraepithelial neoplasia (PanIN) (79.3%, 23/29 and 62.8%, 22/35, respectively). 18794910 0 71 theme pancreatic 60:69 arg1 cancer 71:76 pancreatic cancer 60:76 pancreatic cancer 60:76 RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. 18794910 2 72 theme protein 251:257 arg1 ligase 313:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 72 theme protein 251:257 arg1 RNF13 259:263 a RING-finger-containing protein RNF13 226:263 a RING-finger-containing protein RNF13 226:263 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 3 73 dep % 574:574 arg1 30/72 577:581 30/72 577:581 30/72 577:581 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 2 74 theme ubiquitin 367:375 arg1 activity 384:391 the ubiquitin ligase activity 363:391 the ubiquitin ligase activity 363:391 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 3 75 from analysis 414:421 arg1 patients 507:514 72 patients 504:514 72 patients 504:514 Immunohistochemical analysis of pancreatic ductal adenocarcinoma (PDAC) and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples (41.7%, 30/72), and its expression was significantly associated with histological grading (P = 0.024). 18794910 2 76 theme RING-finger-containing 228:249 arg1 ligase 313:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 an ER/Golgi membrane-associated E3 ubiquitin ligase 268:318 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 18794910 2 76 theme RING-finger-containing 228:249 arg1 RNF13 259:263 a RING-finger-containing protein RNF13 226:263 a RING-finger-containing protein RNF13 226:263 In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. 29429898 5 0 theme α 793:793 arg1 helix 795:799 the C-terminal α helix 778:799 the C-terminal α helix in scKMO and shifts of domain III in pfKMO 778:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 4 1 theme bond 499:502 arg1 network 504:510 the hydrogen bond network 486:510 the hydrogen bond network 486:510 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 6 2 theme Ro 890:891 arg1 61-8048 893:899 Ro 61-8048 890:899 Ro 61-8048 890:899 Interestingly, Ro 61-8048 has two different binding modes. 29429898 4 3 theme flavin 601:606 arg1 reduction 608:616 flavin reduction 601:616 flavin reduction in KMO 601:623 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 3 4 theme human 347:351 arg1 KMO 353:355 human KMO 347:355 human KMO 347:355 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 4 5 theme flavin 521:526 arg1 reduction 528:536 flavin reduction 521:536 flavin reduction in p-hydroxybenzoate hydroxylase 521:569 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 5 6 from scKMO 804:808 arg1 disorder 766:773 disorder 766:773 disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO 766:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 5 6 from scKMO 804:808 arg1 pfKMO 838:842 pfKMO 838:842 pfKMO 838:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 7 7 theme a non-substrate 985:999 arg1 effector 1001:1008 a non-substrate effector 985:1008 a non-substrate effector in pfKMO 985:1017 It acts as a competitive inhibitor in scKMO and as a non-substrate effector in pfKMO. 29429898 4 8 from reduction 608:616 arg1 KMO 621:623 KMO 621:623 KMO 621:623 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 8 9 theme inhibitors 1138:1147 arg1 design 1124:1129 structure-based drug design 1103:1129 structure-based drug design of KMO inhibitors 1103:1147 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 3 10 theme crystal 361:367 arg1 structures 369:378 crystal structures 361:378 crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048 361:464 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 5 11 theme Conformational 639:652 arg1 changes 654:660 Conformational changes 639:660 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors 639:756 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 1 12 theme neurodegenerative 184:200 arg1 disorders 202:210 neurodegenerative disorders 184:210 neurodegenerative disorders 184:210 Kynurenine 3-monooxygenase (KMO) inhibitors have been developed for the treatment of neurodegenerative disorders. 29429898 3 13 theme structures 369:378 arg1 structure 334:342 the structure 330:342 the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048 330:464 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 4 14 theme hydrogen 490:497 arg1 network 504:510 the hydrogen bond network 486:510 the hydrogen bond network 486:510 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 0 15 theme Peroxide 48:55 arg1 Production 57:66 Inhibitor-Induced Hydrogen Peroxide Production 21:66 Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase 21:96 Structural Basis for Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase. 29429898 1 16 theme Kynurenine 99:108 arg1 KMO 127:129 KMO 127:129 KMO 127:129 Kynurenine 3-monooxygenase (KMO) inhibitors have been developed for the treatment of neurodegenerative disorders. 29429898 1 16 theme Kynurenine 99:108 arg1 3-monooxygenase 110:124 Kynurenine 3-monooxygenase 99:124 Kynurenine 3-monooxygenase (KMO) inhibitors 99:141 Kynurenine 3-monooxygenase (KMO) inhibitors have been developed for the treatment of neurodegenerative disorders. 29429898 8 17 theme structure-based 1103:1117 arg1 design 1124:1129 structure-based drug design 1103:1129 structure-based drug design of KMO inhibitors 1103:1147 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 1 18 theme disorders 202:210 arg1 treatment 171:179 the treatment 167:179 the treatment of neurodegenerative disorders 167:210 Kynurenine 3-monooxygenase (KMO) inhibitors have been developed for the treatment of neurodegenerative disorders. 29429898 5 19 theme helix 795:799 arg1 disorder 766:773 disorder 766:773 disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO 766:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 8 20 theme KMO 1083:1085 arg1 cycle 1074:1078 the catalytic cycle 1060:1078 the catalytic cycle of KMO 1060:1085 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 7 21 theme competitive 947:957 arg1 inhibitor 959:967 a competitive inhibitor 945:967 a competitive inhibitor in scKMO and as a non-substrate effector in pfKMO 945:1017 It acts as a competitive inhibitor in scKMO and as a non-substrate effector in pfKMO. 29429898 1 22 theme 3-monooxygenase 110:124 arg1 inhibitors 132:141 Kynurenine 3-monooxygenase (KMO) inhibitors 99:141 Kynurenine 3-monooxygenase (KMO) inhibitors 99:141 Kynurenine 3-monooxygenase (KMO) inhibitors have been developed for the treatment of neurodegenerative disorders. 29429898 7 23 from effector 1001:1008 arg1 pfKMO 1013:1017 pfKMO 1013:1017 pfKMO 1013:1017 It acts as a competitive inhibitor in scKMO and as a non-substrate effector in pfKMO. 29429898 0 24 theme Structural 0:9 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis for Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase 0:96 Structural Basis for Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase. 29429898 0 25 theme Inhibitor-Induced 21:37 arg1 Production 57:66 Inhibitor-Induced Hydrogen Peroxide Production 21:66 Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase 21:96 Structural Basis for Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase. 29429898 5 26 theme domain 824:829 arg1 shifts 814:819 shifts 814:819 shifts of domain III in pfKMO 814:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 5 26 theme domain 824:829 arg1 scKMO 804:808 scKMO 804:808 scKMO 804:808 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 3 27 theme cerevisiae 397:406 arg1 KMO 446:448 Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO 383:448 Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO 383:448 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 3 28 with KMO 353:355 arg1 Ro 455:456 Ro 61-8048 455:464 Ro 61-8048 455:464 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 8 29 theme cycle 1074:1078 arg1 understanding 1043:1055 understanding 1043:1055 understanding of the catalytic cycle of KMO 1043:1085 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 8 29 theme cycle 1074:1078 arg1 insight 1091:1097 insight 1091:1097 insight for structure-based drug design of KMO inhibitors 1091:1147 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 5 30 from disorder 766:773 arg1 shifts 814:819 shifts 814:819 shifts of domain III in pfKMO 814:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 5 30 from disorder 766:773 arg1 scKMO 804:808 scKMO 804:808 scKMO 804:808 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 3 31 theme fluorescens  429:440 arg1 KMO 446:448 Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO 383:448 Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO 383:448 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 5 32 theme non-substrate 734:746 arg1 effectors 748:756 the flavin and substrate or non-substrate effectors 706:756 effectors 748:756 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 2 33 theme production 270:279 arg1 unknown 303:309 unknown 303:309 unknown 303:309 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 2 33 theme production 270:279 arg1 mechanisms 217:226 The mechanisms 213:226 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors 213:297 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 6 34 contain has 901:903 arg2 modes 927:931 two different binding modes 905:931 two different binding modes 905:931 Interestingly, Ro 61-8048 has two different binding modes. 29429898 6 34 contain has 901:903 arg1 61-8048 893:899 Ro 61-8048 890:899 Ro 61-8048 890:899 Interestingly, Ro 61-8048 has two different binding modes. 29429898 0 35 theme Hydrogen 39:46 arg1 Peroxide 48:55 Hydrogen Peroxide 39:55 Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase 21:96 Structural Basis for Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase. 29429898 4 36 theme Proton 467:472 arg1 transfer 474:481 Proton transfer 467:481 Proton transfer in the hydrogen bond network 467:510 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 2 37 theme peroxide 261:268 arg1 production 270:279 hydrogen peroxide production 252:279 hydrogen peroxide production 252:279 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 8 38 theme catalytic 1064:1072 arg1 cycle 1074:1078 the catalytic cycle 1060:1078 the catalytic cycle of KMO 1060:1085 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 5 39 theme flavin 857:862 arg1 reduction 864:872 flavin reduction 857:872 flavin reduction 857:872 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 3 40 theme KMO 446:448 arg1 KMO 353:355 human KMO 347:355 human KMO 347:355 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 3 40 theme KMO 446:448 arg1 structures 369:378 crystal structures 361:378 crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048 361:464 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 2 41 theme hydrogen 252:259 arg1 peroxide 261:268 hydrogen peroxide 252:268 hydrogen peroxide production 252:279 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 4 42 from reduction 528:536 arg1 hydroxylase 559:569 p-hydroxybenzoate hydroxylase 541:569 p-hydroxybenzoate hydroxylase 541:569 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 8 43 theme drug 1119:1122 arg1 design 1124:1129 structure-based drug design 1103:1129 structure-based drug design of KMO inhibitors 1103:1147 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 2 44 theme reduction 238:246 arg1 unknown 303:309 unknown 303:309 unknown 303:309 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 2 44 theme reduction 238:246 arg1 mechanisms 217:226 The mechanisms 213:226 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors 213:297 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 5 45 from helix 795:799 arg1 shifts 814:819 shifts 814:819 shifts of domain III in pfKMO 814:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 5 45 from helix 795:799 arg1 scKMO 804:808 scKMO 804:808 scKMO 804:808 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 5 46 from shifts 814:819 arg1 disorder 766:773 disorder 766:773 disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO 766:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 5 46 from shifts 814:819 arg1 pfKMO 838:842 pfKMO 838:842 pfKMO 838:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 4 47 from transfer 474:481 arg1 network 504:510 the hydrogen bond network 486:510 the hydrogen bond network 486:510 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 4 48 theme p-hydroxybenzoate 541:557 arg1 hydroxylase 559:569 p-hydroxybenzoate hydroxylase 541:569 p-hydroxybenzoate hydroxylase 541:569 Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. 29429898 8 49 theme KMO 1134:1136 arg1 inhibitors 1138:1147 KMO inhibitors 1134:1147 KMO inhibitors 1134:1147 These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors. 29429898 2 50 theme flavin 231:236 arg1 reduction 238:246 flavin reduction 231:246 flavin reduction 231:246 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 3 51 with structures 369:378 arg1 Ro 455:456 Ro 61-8048 455:464 Ro 61-8048 455:464 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 6 52 theme binding 919:925 arg1 modes 927:931 two different binding modes 905:931 two different binding modes 905:931 Interestingly, Ro 61-8048 has two different binding modes. 29429898 3 53 theme KMO 353:355 arg1 structure 334:342 the structure 330:342 the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048 330:464 Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. 29429898 2 54 theme KMO 284:286 arg1 inhibitors 288:297 KMO inhibitors 284:297 KMO inhibitors 284:297 The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. 29429898 7 55 from inhibitor 959:967 arg1 scKMO 972:976 scKMO 972:976 scKMO 972:976 It acts as a competitive inhibitor in scKMO and as a non-substrate effector in pfKMO. 29429898 5 56 theme C-terminal 782:791 arg1 helix 795:799 the C-terminal α helix 778:799 the C-terminal α helix in scKMO and shifts of domain III in pfKMO 778:842 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 29429898 6 57 theme different 909:917 arg1 modes 927:931 two different binding modes 905:931 two different binding modes 905:931 Interestingly, Ro 61-8048 has two different binding modes. 29429898 0 58 theme Kynurenine 71:80 arg1 3-Monooxygenase 82:96 Kynurenine 3-Monooxygenase 71:96 Kynurenine 3-Monooxygenase 71:96 Structural Basis for Inhibitor-Induced Hydrogen Peroxide Production by Kynurenine 3-Monooxygenase. 29429898 5 59 theme π-π 666:668 arg1 interactions 670:681 π-π interactions 666:681 π-π interactions between the loop above the flavin and substrate or non-substrate effectors 666:756 Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. 30639100 3 0 theme GPCRs 685:689 arg1 characteristic 657:670 characteristic 657:670 characteristic 657:670 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 4 1 theme spanning from 752:764 arg1 remodeling 826:835 remodeling 826:835 remodeling the ligand-binding cavity 826:861 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 4 1 theme spanning from 752:764 arg1 interface 742:750 an extensive interface 729:750 an extensive interface spanning from the receptor core to its extracellular face and N terminus 729:823 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 1 2 theme important 213:221 arg1 model 223:227 an important model 210:227 an important model for studies of G-protein-coupled receptor (GPCR) signaling 210:286 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 2 3 theme crystal 441:447 arg1 structure 449:457 the crystal structure 437:457 the crystal structure of active-state human AT1R 437:484 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 3 4 theme characteristic 657:670 arg1 changes 649:655 the large conformational changes 624:655 the large conformational changes characteristic of activated GPCRs 624:689 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 4 5 theme ligand-binding 841:854 arg1 cavity 856:861 the ligand-binding cavity 837:861 remodeling the ligand-binding cavity 826:861 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 4 6 theme receptor 770:777 arg1 core 779:782 the receptor core 766:782 the receptor core 766:782 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 5 7 theme key 989:991 arg1 sites 993:997 several key sites 981:997 several key sites 981:997 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 2 8 dep determined 426:435 arg1 bound 486:490 bound 486:490 determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity 426:539 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 4 9 theme extensive 732:740 arg1 remodeling 826:835 remodeling 826:835 remodeling the ligand-binding cavity 826:861 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 4 9 theme extensive 732:740 arg1 interface 742:750 an extensive interface 729:750 an extensive interface spanning from the receptor core to its extracellular face and N terminus 729:823 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 5 10 from sites 993:997 arg1 GPCRs 972:976 other GPCRs 966:976 other GPCRs at several key sites 966:997 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 2 11 theme AT1R 481:484 arg1 structure 449:457 the crystal structure 437:457 the crystal structure of active-state human AT1R 437:484 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 4 12 dep core 779:782 arg1 face 805:808 its extracellular face 787:808 its extracellular face 787:808 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 4 12 dep core 779:782 arg1 terminus 816:823 N terminus 814:823 N terminus 814:823 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 2 13 theme nanobody 358:365 arg1 fragment 347:354 a single-domain antibody fragment 322:354 a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library 322:420 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 13 theme nanobody 358:365 arg1 " 366:366 "nanobody" 357:366 "nanobody" 357:366 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 14 theme human 475:479 arg1 AT1R 481:484 active-state human AT1R 462:484 active-state human AT1R 462:484 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 15 theme yeast-displayed 398:412 arg1 library 414:420 a synthetic yeast-displayed library 386:420 a synthetic yeast-displayed library 386:420 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 0 16 theme Activation 12:21 arg1 Mechanism 23:31 Distinctive Activation Mechanism 0:31 Distinctive Activation Mechanism for Angiotensin Receptor 0:56 Distinctive Activation Mechanism for Angiotensin Receptor Revealed by a Synthetic Nanobody. 30639100 2 17 theme active-state 462:473 arg1 AT1R 481:484 active-state human AT1R 462:484 active-state human AT1R 462:484 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 1 18 theme critical 147:154 arg1 regulator 156:164 a critical regulator 145:164 a critical regulator of cardiovascular and renal function 145:201 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 18 theme critical 147:154 arg1 receptor 126:133 The angiotensin II (AngII) type 1 receptor 92:133 The angiotensin II (AngII) type 1 receptor (AT1R) 92:140 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 0 19 theme Distinctive 0:10 arg1 Mechanism 23:31 Distinctive Activation Mechanism 0:31 Distinctive Activation Mechanism for Angiotensin Receptor 0:56 Distinctive Activation Mechanism for Angiotensin Receptor Revealed by a Synthetic Nanobody. 30639100 1 20 theme angiotensin 96:106 arg1 regulator 156:164 a critical regulator 145:164 a critical regulator of cardiovascular and renal function 145:201 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 20 theme angiotensin 96:106 arg1 AT1R 136:139 AT1R 136:139 AT1R 136:139 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 20 theme angiotensin 96:106 arg1 receptor 126:133 The angiotensin II (AngII) type 1 receptor 92:133 The angiotensin II (AngII) type 1 receptor (AT1R) 92:140 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 2 21 theme synthetic 388:396 arg1 library 414:420 a synthetic yeast-displayed library 386:420 a synthetic yeast-displayed library 386:420 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 1 22 theme G-protein-coupled 244:260 arg1 receptor 262:269 G-protein-coupled receptor 244:269 G-protein-coupled receptor (GPCR) signaling 244:286 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 22 theme G-protein-coupled 244:260 arg1 GPCR 272:275 GPCR 272:275 GPCR 272:275 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 5 23 theme allosteric 1030:1039 arg1 mechanisms 1041:1050 allosteric mechanisms 1030:1050 allosteric mechanisms 1030:1050 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 6 24 theme biased 1145:1150 arg1 signaling 1152:1160 full or biased signaling 1137:1160 full or biased signaling 1137:1160 Our structure provides insight into how AngII and its analogs stimulate full or biased signaling, respectively. 30639100 1 25 theme receptor 262:269 arg1 signaling 278:286 G-protein-coupled receptor (GPCR) signaling 244:286 G-protein-coupled receptor (GPCR) signaling 244:286 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 6 26 theme full 1137:1140 arg1 signaling 1152:1160 full or biased signaling 1137:1160 full or biased signaling 1137:1160 Our structure provides insight into how AngII and its analogs stimulate full or biased signaling, respectively. 30639100 5 27 theme other 966:970 arg1 GPCRs 972:976 other GPCRs 966:976 other GPCRs at several key sites 966:997 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 1 28 theme cardiovascular 169:182 arg1 function 194:201 cardiovascular and renal function 169:201 cardiovascular and renal function 169:201 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 2 29 theme AngII 498:502 arg1 analog 504:509 an AngII analog 495:509 an AngII analog with partial agonist activity 495:539 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 0 30 theme Angiotensin 37:47 arg1 Receptor 49:56 Angiotensin Receptor 37:56 Angiotensin Receptor 37:56 Distinctive Activation Mechanism for Angiotensin Receptor Revealed by a Synthetic Nanobody. 30639100 2 31 with receptor 308:315 arg1 fragment 347:354 a single-domain antibody fragment 322:354 a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library 322:420 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 31 with receptor 308:315 arg1 " 366:366 "nanobody" 357:366 "nanobody" 357:366 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 4 32 theme extracellular 791:803 arg1 face 805:808 its extracellular face 787:808 its extracellular face 787:808 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 1 33 theme renal 188:192 arg1 function 194:201 cardiovascular and renal function 169:201 cardiovascular and renal function 169:201 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 4 34 theme N 814:814 arg1 terminus 816:823 N terminus 814:823 N terminus 814:823 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 1 35 theme signaling 278:286 arg1 studies 233:239 studies 233:239 studies of G-protein-coupled receptor (GPCR) signaling 233:286 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 3 36 theme conformational 634:647 arg1 changes 649:655 the large conformational changes 624:655 the large conformational changes characteristic of activated GPCRs 624:689 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 3 37 theme transducer 593:602 arg1 pocket 604:609 the receptor's intracellular transducer pocket 564:609 the receptor's intracellular transducer pocket 564:609 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 2 38 theme agonist 524:530 arg1 activity 532:539 partial agonist activity 516:539 partial agonist activity 516:539 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 5 39 theme mechanisms 1041:1050 arg1 diversity 1017:1025 the diversity 1013:1025 the diversity of allosteric mechanisms among GPCRs 1013:1062 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 5 40 theme several 981:987 arg1 sites 993:997 several key sites 981:997 several key sites 981:997 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 1 41 theme type 119:122 arg1 regulator 156:164 a critical regulator 145:164 a critical regulator of cardiovascular and renal function 145:201 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 41 theme type 119:122 arg1 AT1R 136:139 AT1R 136:139 AT1R 136:139 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 41 theme type 119:122 arg1 receptor 126:133 The angiotensin II (AngII) type 1 receptor 92:133 The angiotensin II (AngII) type 1 receptor (AT1R) 92:140 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 42 theme function 194:201 arg1 model 223:227 an important model 210:227 an important model for studies of G-protein-coupled receptor (GPCR) signaling 210:286 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 42 theme function 194:201 arg1 regulator 156:164 a critical regulator 145:164 a critical regulator of cardiovascular and renal function 145:201 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 1 42 theme function 194:201 arg1 receptor 126:133 The angiotensin II (AngII) type 1 receptor 92:133 The angiotensin II (AngII) type 1 receptor (AT1R) 92:140 The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. 30639100 2 43 theme partial 516:522 arg1 activity 532:539 partial agonist activity 516:539 partial agonist activity 516:539 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 3 44 theme intracellular 579:591 arg1 pocket 604:609 the receptor's intracellular transducer pocket 564:609 the receptor's intracellular transducer pocket 564:609 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 5 45 theme conformational 908:921 arg1 changes 923:929 conformational changes 908:929 conformational changes 908:929 Remarkably, the mechanism used to propagate conformational changes through the receptor diverges from other GPCRs at several key sites, highlighting the diversity of allosteric mechanisms among GPCRs. 30639100 0 46 theme Synthetic 72:80 arg1 Nanobody 82:89 a Synthetic Nanobody 70:89 a Synthetic Nanobody 70:89 Distinctive Activation Mechanism for Angiotensin Receptor Revealed by a Synthetic Nanobody. 30639100 3 47 theme large 628:632 arg1 changes 649:655 the large conformational changes 624:655 the large conformational changes characteristic of activated GPCRs 624:689 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 2 48 theme antibody 338:345 arg1 fragment 347:354 a single-domain antibody fragment 322:354 a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library 322:420 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 48 theme antibody 338:345 arg1 " 366:366 "nanobody" 357:366 "nanobody" 357:366 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 49 with analog 504:509 arg1 activity 532:539 partial agonist activity 516:539 partial agonist activity 516:539 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 4 50 dep remodeling 826:835 arg1 cavity 856:861 the ligand-binding cavity 837:861 remodeling the ligand-binding cavity 826:861 The peptide engages the AT1R through an extensive interface spanning from the receptor core to its extracellular face and N terminus, remodeling the ligand-binding cavity. 30639100 3 51 theme activated 675:683 arg1 GPCRs 685:689 activated GPCRs 675:689 activated GPCRs 675:689 The nanobody binds to the receptor's intracellular transducer pocket, stabilizing the large conformational changes characteristic of activated GPCRs. 30639100 2 52 theme single-domain 324:336 arg1 fragment 347:354 a single-domain antibody fragment 322:354 a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library 322:420 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 30639100 2 52 theme single-domain 324:336 arg1 " 366:366 "nanobody" 357:366 "nanobody" 357:366 By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered using a synthetic yeast-displayed library, we determined the crystal structure of active-state human AT1R bound to an AngII analog with partial agonist activity. 28292932 7 0 theme new 1208:1210 arg1 light 1212:1216 new light 1208:1216 new light 1208:1216 These results shed new light on substrate specificity determinants within the NPP enzyme family. 28292932 1 1 theme initial 157:163 arg1 degradation 165:175 its initial degradation 153:175 its initial degradation 153:175 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 0 2 theme substrate 80:88 arg1 recognition 90:100 substrate recognition 80:100 substrate recognition 80:100 Crystal structure of the human alkaline sphingomyelinase provides insights into substrate recognition. 28292932 5 3 theme choline 885:891 arg1 moiety 893:898 the choline moiety 881:898 the choline moiety of its substrates 881:916 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 6 4 theme Mutational 983:992 arg1 analysis 994:1001 Mutational analysis 983:1001 Mutational analysis 983:1001 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 6 5 theme patch 1094:1098 arg1 loop-that 1125:1133 a unique hydrophobic loop-that 1104:1133 a unique hydrophobic loop-that 1104:1133 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 6 5 theme patch 1094:1098 arg1 surface 1060:1066 the surface 1056:1066 the surface of the protein-a cationic patch 1056:1098 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 5 6 theme aromatic 939:946 arg1 box 948:950 an NPP7-specific aromatic box 922:950 an NPP7-specific aromatic box composed of tyrosine residues 922:980 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 7 7 theme NPP 1267:1269 arg1 family 1278:1283 the NPP enzyme family 1263:1283 the NPP enzyme family 1263:1283 These results shed new light on substrate specificity determinants within the NPP enzyme family. 28292932 6 8 theme salt 1174:1177 arg1 micelles 1179:1186 bile salt micelles 1169:1186 bile salt micelles 1169:1186 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 2 9 theme family 365:370 arg1 members 377:383 the members 373:383 the members 373:383 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 3 10 theme only 483:486 arg1 paralog 488:494 the only paralog 479:494 the only paralog that can cleave SM 479:513 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 3 10 theme only 483:486 arg1 NPP7 471:474 NPP7 471:474 NPP7 471:474 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 2 11 theme NPP 360:362 arg1 family 365:370 the nucleotide pyrophosphatase/phosphodiesterase (NPP) family 310:370 the nucleotide pyrophosphatase/phosphodiesterase (NPP) family 310:370 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 4 12 theme reaction 757:764 arg1 product 766:772 a reaction product 755:772 a reaction product 755:772 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 4 12 theme reaction 757:764 arg1 phosphocholine 739:752 phosphocholine 739:752 phosphocholine 739:752 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 6 13 theme protein-a 1075:1083 arg1 patch 1094:1098 the protein-a cationic patch 1071:1098 the protein-a cationic patch 1071:1098 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 6 14 theme hydrophobic 1113:1123 arg1 loop-that 1125:1133 a unique hydrophobic loop-that 1104:1133 a unique hydrophobic loop-that 1104:1133 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 6 15 from features 1044:1051 arg1 loop-that 1125:1133 a unique hydrophobic loop-that 1104:1133 a unique hydrophobic loop-that 1104:1133 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 6 15 from features 1044:1051 arg1 surface 1060:1066 the surface 1056:1066 the surface of the protein-a cationic patch 1056:1098 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 0 16 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human alkaline sphingomyelinase 0:55 Crystal structure of the human alkaline sphingomyelinase provides insights into substrate recognition. 28292932 6 17 theme cationic 1085:1092 arg1 patch 1094:1098 the protein-a cationic patch 1071:1098 the protein-a cationic patch 1071:1098 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 5 18 theme other 848:852 arg1 NPPs 854:857 other NPPs 848:857 other NPPs 848:857 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 2 19 theme related 452:458 arg1 molecules 460:468 other related molecules 446:468 other related molecules 446:468 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 7 20 theme enzyme 1271:1276 arg1 family 1278:1283 the NPP enzyme family 1263:1283 the NPP enzyme family 1263:1283 These results shed new light on substrate specificity determinants within the NPP enzyme family. 28292932 4 21 theme recognition 655:665 arg1 mechanism 632:640 the mechanism 628:640 the mechanism of substrate recognition 628:665 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 2 22 theme other 446:450 arg1 molecules 460:468 other related molecules 446:468 other related molecules 446:468 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 3 23 theme bile 558:561 arg1 salts 563:567 bile salts 558:567 bile salts 558:567 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 3 23 theme bile 558:561 arg1 class 572:576 a class 570:576 a class of physiological anionic detergents 570:612 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 4 24 theme substrate 645:653 arg1 recognition 655:665 substrate recognition 645:665 substrate recognition 645:665 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 5 25 theme substrates 907:916 arg1 moiety 893:898 the choline moiety 881:898 the choline moiety of its substrates 881:916 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 5 26 theme NPP7-specific 925:937 arg1 box 948:950 an NPP7-specific aromatic box 922:950 an NPP7-specific aromatic box composed of tyrosine residues 922:980 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 4 27 theme alk-SMase 713:721 arg1 structure 694:702 the crystal structure 682:702 the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product 682:772 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 6 28 theme bile 1169:1172 arg1 micelles 1179:1186 bile salt micelles 1169:1186 bile salt micelles 1169:1186 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 1 29 theme dietary 117:123 arg1 SM 140:141 SM 140:141 SM 140:141 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 1 29 theme dietary 117:123 arg1 sphingomyelin 125:137 dietary sphingomyelin 117:137 dietary sphingomyelin (SM) 117:142 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 0 30 theme alkaline 31:38 arg1 sphingomyelinase 40:55 the human alkaline sphingomyelinase 21:55 the human alkaline sphingomyelinase 21:55 Crystal structure of the human alkaline sphingomyelinase provides insights into substrate recognition. 28292932 1 31 dep sphingomyelinase 246:261 arg1 alk-SMase 264:272 alk-SMase 264:272 alk-SMase 264:272 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 1 31 dep sphingomyelinase 246:261 arg1 ENPP7 281:285 ENPP7 281:285 ENPP7 281:285 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 1 31 dep sphingomyelinase 246:261 arg1 NPP7 275:278 NPP7 275:278 NPP7 275:278 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 0 32 theme human 25:29 arg1 sphingomyelinase 40:55 the human alkaline sphingomyelinase 21:55 the human alkaline sphingomyelinase 21:55 Crystal structure of the human alkaline sphingomyelinase provides insights into substrate recognition. 28292932 6 33 theme enzymatic 1007:1015 arg1 assays 1026:1031 enzymatic activity assays 1007:1031 enzymatic activity assays 1007:1031 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 2 34 theme nucleoside 404:413 arg1 phosphates 415:424 nucleoside phosphates 404:424 nucleoside phosphates 404:424 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 4 35 with complex 726:732 arg1 product 766:772 a reaction product 755:772 a reaction product 755:772 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 4 35 with complex 726:732 arg1 phosphocholine 739:752 phosphocholine 739:752 phosphocholine 739:752 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 5 36 theme catalytic 805:813 arg1 center 815:820 catalytic center 805:820 catalytic center 805:820 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 3 37 theme salts 563:567 arg1 presence 546:553 the presence 542:553 the presence of bile salts, a class of physiological anionic detergents 542:612 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 1 38 theme sphingomyelin 125:137 arg1 Absorption 103:112 Absorption 103:112 Absorption of dietary sphingomyelin (SM) 103:142 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 0 39 theme sphingomyelinase 40:55 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human alkaline sphingomyelinase 0:55 Crystal structure of the human alkaline sphingomyelinase provides insights into substrate recognition. 28292932 4 40 theme crystal 686:692 arg1 structure 694:702 the crystal structure 682:702 the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product 682:772 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 4 41 theme human 707:711 arg1 alk-SMase 713:721 human alk-SMase 707:721 human alk-SMase 707:721 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 3 42 theme anionic 595:601 arg1 detergents 603:612 physiological anionic detergents 581:612 physiological anionic detergents 581:612 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 6 43 theme activity 1017:1024 arg1 assays 1026:1031 enzymatic activity assays 1007:1031 enzymatic activity assays 1007:1031 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 2 44 theme pyrophosphatase/phosphodiesterase 325:357 arg1 family 365:370 the nucleotide pyrophosphatase/phosphodiesterase (NPP) family 310:370 the nucleotide pyrophosphatase/phosphodiesterase (NPP) family 310:370 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 6 45 theme unique 1106:1111 arg1 loop-that 1125:1133 a unique hydrophobic loop-that 1104:1133 a unique hydrophobic loop-that 1104:1133 Mutational analysis and enzymatic activity assays identified features on the surface of the protein-a cationic patch and a unique hydrophobic loop-that are essential for accessing SM in bile salt micelles. 28292932 1 46 theme intestinal 219:228 arg1 sphingomyelinase 246:261 the intestinal enzyme alkaline sphingomyelinase 215:261 the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7) 215:286 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 5 47 theme tyrosine 964:971 arg1 residues 973:980 tyrosine residues 964:980 tyrosine residues 964:980 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 3 48 theme detergents 603:612 arg1 salts 563:567 bile salts 558:567 bile salts 558:567 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 3 48 theme detergents 603:612 arg1 class 572:576 a class 570:576 a class of physiological anionic detergents 570:612 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 2 49 theme nucleotide 314:323 arg1 family 365:370 the nucleotide pyrophosphatase/phosphodiesterase (NPP) family 310:370 the nucleotide pyrophosphatase/phosphodiesterase (NPP) family 310:370 alk-SMase belongs to the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, the members of which hydrolyze nucleoside phosphates, phospholipids, and other related molecules. 28292932 4 50 from structure 694:702 arg1 complex 726:732 complex 726:732 complex with phosphocholine, a reaction product 726:772 To elucidate the mechanism of substrate recognition, we determined the crystal structure of human alk-SMase in complex with phosphocholine, a reaction product. 28292932 1 51 theme enzyme 230:235 arg1 sphingomyelinase 246:261 the intestinal enzyme alkaline sphingomyelinase 215:261 the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7) 215:286 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 1 52 theme alkaline 237:244 arg1 sphingomyelinase 246:261 the intestinal enzyme alkaline sphingomyelinase 215:261 the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7) 215:286 Absorption of dietary sphingomyelin (SM) requires its initial degradation into ceramide, a process catalyzed by the intestinal enzyme alkaline sphingomyelinase (alk-SMase, NPP7, ENPP7). 28292932 7 53 theme substrate 1221:1229 arg1 determinants 1243:1254 substrate specificity determinants 1221:1254 substrate specificity determinants within the NPP enzyme family 1221:1283 These results shed new light on substrate specificity determinants within the NPP enzyme family. 28292932 3 54 theme physiological 581:593 arg1 detergents 603:612 physiological anionic detergents 581:612 physiological anionic detergents 581:612 NPP7 is the only paralog that can cleave SM, and its activity requires the presence of bile salts, a class of physiological anionic detergents. 28292932 5 55 theme overall 788:794 arg1 fold 796:799 the overall fold 784:799 the overall fold 784:799 Although the overall fold and catalytic center are conserved relative to other NPPs, alk-SMase recognizes the choline moiety of its substrates via an NPP7-specific aromatic box composed of tyrosine residues. 28292932 7 56 theme specificity 1231:1241 arg1 determinants 1243:1254 substrate specificity determinants 1221:1254 substrate specificity determinants within the NPP enzyme family 1221:1283 These results shed new light on substrate specificity determinants within the NPP enzyme family. 17196528 0 0 theme TrkA 83:86 arg1 receptors 96:104 the TrkA and p75 receptors 79:104 the TrkA and p75 receptors 79:104 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 1 1 theme complex 314:320 arg1 formation 278:286 formation 278:286 formation of a ternary TrkA/NGF/p75 complex 278:320 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 1 2 theme distinct 152:159 arg1 TrkA 186:189 TrkA 186:189 TrkA 186:189 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 1 2 theme distinct 152:159 arg1 receptors 175:183 two structurally distinct transmembrane receptors 135:183 two structurally distinct transmembrane receptors 135:183 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 1 2 theme distinct 152:159 arg1 p75 195:197 p75 195:197 p75 195:197 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 2 3 theme TrkA 469:472 arg1 TrkA 469:472 TrkA 469:472 TrkA 469:472 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 2 3 theme TrkA 469:472 arg1 domain 459:464 a complete extracellular domain 434:464 a complete extracellular domain of TrkA complexed with NGF 434:491 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 1 4 theme transmembrane 161:173 arg1 TrkA 186:189 TrkA 186:189 TrkA 186:189 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 1 4 theme transmembrane 161:173 arg1 receptors 175:183 two structurally distinct transmembrane receptors 135:183 two structurally distinct transmembrane receptors 135:183 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 1 4 theme transmembrane 161:173 arg1 p75 195:197 p75 195:197 p75 195:197 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 0 5 theme p75 92:94 arg1 receptors 96:104 the TrkA and p75 receptors 79:104 the TrkA and p75 receptors 79:104 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 3 6 theme crab-shaped 516:526 arg1 structure 545:553 a crab-shaped homodimeric TrkA structure 514:553 a crab-shaped homodimeric TrkA structure 514:553 The complex reveals a crab-shaped homodimeric TrkA structure, but a mechanism for p75 coordination is not obvious. 17196528 1 7 theme high-affinity 238:250 arg1 site 265:268 a "high-affinity" NGF binding site 235:268 a "high-affinity" NGF binding site 235:268 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 4 8 theme interaction 740:750 arg1 system 752:757 a beta-gal protein-protein interaction system 713:757 a beta-gal protein-protein interaction system 713:757 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 3 9 theme homodimeric 528:538 arg1 structure 545:553 a crab-shaped homodimeric TrkA structure 514:553 a crab-shaped homodimeric TrkA structure 514:553 The complex reveals a crab-shaped homodimeric TrkA structure, but a mechanism for p75 coordination is not obvious. 17196528 1 10 theme " 251:251 arg1 site 265:268 a "high-affinity" NGF binding site 235:268 a "high-affinity" NGF binding site 235:268 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 4 11 theme mammalian 690:698 arg1 cells 700:704 intact mammalian cells 683:704 intact mammalian cells 683:704 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 7 12 theme adaptor 1058:1064 arg1 molecules 1066:1074 shared adaptor molecules 1051:1074 shared adaptor molecules 1051:1074 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 0 13 with interactions 61:72 arg1 receptors 96:104 the TrkA and p75 receptors 79:104 the TrkA and p75 receptors 79:104 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 7 14 theme extracellular 1104:1116 arg1 interactions 1118:1129 direct extracellular interactions 1097:1129 direct extracellular interactions 1097:1129 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 0 15 theme nerve 41:45 arg1 interactions 61:72 nerve growth factor interactions 41:72 nerve growth factor interactions with the TrkA and p75 receptors 41:104 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 2 16 theme high-affinity 360:372 arg1 site 374:377 the high-affinity site 356:377 the high-affinity site 356:377 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 6 17 theme TrkA/p75 921:928 arg1 interaction 930:940 a direct TrkA/p75 interaction 912:940 a direct TrkA/p75 interaction 912:940 We find no evidence for a direct TrkA/p75 interaction. 17196528 4 18 theme TrkA 666:669 arg1 heterodimerization 629:646 the heterodimerization 625:646 the heterodimerization of membrane-bound TrkA and p75 625:677 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 0 19 theme Structural 0:9 arg1 insights 27:34 Structural and mechanistic insights 0:34 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 0:105 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 1 20 theme NGF 253:255 arg1 site 265:268 a "high-affinity" NGF binding site 235:268 a "high-affinity" NGF binding site 235:268 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 7 21 theme shared 1051:1056 arg1 molecules 1066:1074 shared adaptor molecules 1051:1074 shared adaptor molecules 1051:1074 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 4 22 theme membrane-bound 651:664 arg1 TrkA 666:669 membrane-bound TrkA 651:669 membrane-bound TrkA 651:669 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 2 23 theme three-dimensional 403:419 arg1 structure 421:429 the three-dimensional structure 399:429 the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF 399:491 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 1 24 theme binding 257:263 arg1 site 265:268 a "high-affinity" NGF binding site 235:268 a "high-affinity" NGF binding site 235:268 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 7 25 theme signaling 1025:1033 arg1 pathways 1035:1042 downstream signaling pathways 1014:1042 downstream signaling pathways 1014:1042 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 3 26 theme TrkA 540:543 arg1 structure 545:553 a crab-shaped homodimeric TrkA structure 514:553 a crab-shaped homodimeric TrkA structure 514:553 The complex reveals a crab-shaped homodimeric TrkA structure, but a mechanism for p75 coordination is not obvious. 17196528 0 27 theme mechanistic 15:25 arg1 insights 27:34 Structural and mechanistic insights 0:34 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 0:105 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 7 28 theme pathways 1035:1042 arg1 convergence 999:1009 convergence 999:1009 convergence of downstream signaling pathways and/or shared adaptor molecules 999:1074 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 4 29 theme beta-gal 715:722 arg1 system 752:757 a beta-gal protein-protein interaction system 713:757 a beta-gal protein-protein interaction system 713:757 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 2 30 theme domain 459:464 arg1 structure 421:429 the three-dimensional structure 399:429 the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF 399:491 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 4 31 theme intact 683:688 arg1 cells 700:704 intact mammalian cells 683:704 intact mammalian cells 683:704 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 5 32 theme preformed 837:845 arg1 oligomer 847:854 a preformed oligomer 835:854 a preformed oligomer that is not dissociated by NGF 835:885 We find that NGF dimerizes TrkA and that p75 exists on the cell surface as a preformed oligomer that is not dissociated by NGF. 17196528 7 33 theme direct 1097:1102 arg1 interactions 1118:1129 direct extracellular interactions 1097:1129 direct extracellular interactions 1097:1129 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 2 34 theme extracellular 445:457 arg1 TrkA 469:472 TrkA 469:472 TrkA 469:472 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 2 34 theme extracellular 445:457 arg1 domain 459:464 a complete extracellular domain 434:464 a complete extracellular domain of TrkA complexed with NGF 434:491 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 7 35 theme downstream 1014:1023 arg1 pathways 1035:1042 downstream signaling pathways 1014:1042 downstream signaling pathways 1014:1042 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 3 36 theme p75 576:578 arg1 coordination 580:591 p75 coordination 576:591 p75 coordination 576:591 The complex reveals a crab-shaped homodimeric TrkA structure, but a mechanism for p75 coordination is not obvious. 17196528 5 37 theme cell 819:822 arg1 surface 824:830 the cell surface 815:830 the cell surface 815:830 We find that NGF dimerizes TrkA and that p75 exists on the cell surface as a preformed oligomer that is not dissociated by NGF. 17196528 0 38 theme factor 54:59 arg1 interactions 61:72 nerve growth factor interactions 41:72 nerve growth factor interactions with the TrkA and p75 receptors 41:104 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 6 39 theme direct 914:919 arg1 interaction 930:940 a direct TrkA/p75 interaction 912:940 a direct TrkA/p75 interaction 912:940 We find no evidence for a direct TrkA/p75 interaction. 17196528 1 40 theme Nerve 107:111 arg1 factor 120:125 Nerve growth factor 107:125 Nerve growth factor 107:125 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 0 41 theme growth 47:52 arg1 interactions 61:72 nerve growth factor interactions 41:72 nerve growth factor interactions with the TrkA and p75 receptors 41:104 Structural and mechanistic insights into nerve growth factor interactions with the TrkA and p75 receptors. 17196528 7 42 theme molecules 1066:1074 arg1 convergence 999:1009 convergence 999:1009 convergence of downstream signaling pathways and/or shared adaptor molecules 999:1074 We propose that TrkA and p75 likely communicate through convergence of downstream signaling pathways and/or shared adaptor molecules, rather than through direct extracellular interactions. 17196528 1 43 theme growth 113:118 arg1 factor 120:125 Nerve growth factor 107:125 Nerve growth factor 107:125 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 1 44 theme ternary 293:299 arg1 complex 314:320 a ternary TrkA/NGF/p75 complex 291:320 a ternary TrkA/NGF/p75 complex 291:320 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 4 45 theme protein-protein 724:738 arg1 system 752:757 a beta-gal protein-protein interaction system 713:757 a beta-gal protein-protein interaction system 713:757 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 4 46 theme p75 675:677 arg1 heterodimerization 629:646 the heterodimerization 625:646 the heterodimerization of membrane-bound TrkA and p75 625:677 We investigated the heterodimerization of membrane-bound TrkA and p75, on intact mammalian cells, using a beta-gal protein-protein interaction system. 17196528 2 47 theme complete 436:443 arg1 TrkA 469:472 TrkA 469:472 TrkA 469:472 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 2 47 theme complete 436:443 arg1 domain 459:464 a complete extracellular domain 434:464 a complete extracellular domain of TrkA complexed with NGF 434:491 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 17196528 1 48 theme TrkA/NGF/p75 301:312 arg1 complex 314:320 a ternary TrkA/NGF/p75 complex 291:320 a ternary TrkA/NGF/p75 complex 291:320 Nerve growth factor engages two structurally distinct transmembrane receptors, TrkA and p75, which have been proposed to create a "high-affinity" NGF binding site through formation of a ternary TrkA/NGF/p75 complex. 17196528 2 49 theme structural 335:344 arg1 basis 346:350 a structural basis 333:350 a structural basis for the high-affinity site 333:377 To define a structural basis for the high-affinity site, we have determined the three-dimensional structure of a complete extracellular domain of TrkA complexed with NGF. 3163150 4 0 theme serine 497:502 arg1 residues 504:511 two serine residues 493:511 two serine residues 493:511 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 3 1 theme half-cystine 233:244 arg1 residues 246:253 Ten half-cystine residues 229:253 Ten half-cystine residues 229:253 Ten half-cystine residues form five disulfide bridges. 3163150 5 2 theme porcine 667:673 arg1 proteinases 644:654 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 2 theme porcine 667:673 arg1 kallikrein 675:684 porcine kallikrein 667:684 porcine kallikrein 667:684 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 3 theme urinary 599:605 arg1 kallikrein 607:616 human urinary kallikrein 593:616 human urinary kallikrein 593:616 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 4 theme kallikrein 607:616 arg1 structure 580:588 the primary structure 568:588 the primary structure of human urinary kallikrein 568:616 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 6 5 dep CDGVLQGVTSWGYVPCGTPNKPSVAVRVLSYVKWIEDTIAENS 938:980 arg1 AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN 778:816 IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN 738:816 IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN 738:816 IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN MSLLENHTRQADEDYSHDLMLLRLTEPADTITDAVKVVE LPTQEPEVGSTCLASGWGSIEPENFSFPDDLQCVDLKIL PNDECEKAHVQKVTDFMLCVGHLEGGKDTCVGDSGGPLM CDGVLQGVTSWGYVPCGTPNKPSVAVRVLSYVKWIEDTIAENS. 3163150 4 6 from present 399:405 arg1 Asn-Xaa-Thr 430:440 the acceptor sequon Asn-Xaa-Thr 410:440 the acceptor sequon Asn-Xaa-Thr (or -Ser) 410:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 6 from present 399:405 arg1 -Ser 446:449 -Ser 446:449 -Ser 446:449 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 7 theme sequon 423:428 arg1 Asn-Xaa-Thr 430:440 the acceptor sequon Asn-Xaa-Thr 410:440 the acceptor sequon Asn-Xaa-Thr (or -Ser) 410:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 7 theme sequon 423:428 arg1 -Ser 446:449 -Ser 446:449 -Ser 446:449 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 5 8 with Comparison 554:563 arg1 that 623:626 that 623:626 that 623:626 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 3 9 theme disulfide 265:273 arg1 bridges 275:281 five disulfide bridges 260:281 five disulfide bridges 260:281 Ten half-cystine residues form five disulfide bridges. 3163150 4 10 theme acceptor 414:421 arg1 Asn-Xaa-Thr 430:440 the acceptor sequon Asn-Xaa-Thr 410:440 the acceptor sequon Asn-Xaa-Thr (or -Ser) 410:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 10 theme acceptor 414:421 arg1 -Ser 446:449 -Ser 446:449 -Ser 446:449 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 11 attach present 399:405 arg2 asparagine 379:388 asparagine 379:388 asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) 379:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 11 attach present 399:405 arg1 Asn-Xaa-Thr 430:440 the acceptor sequon Asn-Xaa-Thr 410:440 the acceptor sequon Asn-Xaa-Thr (or -Ser) 410:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 11 attach present 399:405 arg1 -Ser 446:449 -Ser 446:449 -Ser 446:449 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 5 12 theme other 631:635 arg1 proteinases 644:654 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 12 theme other 631:635 arg1 kallikrein 675:684 porcine kallikrein 667:684 porcine kallikrein 667:684 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 12 theme other 631:635 arg1 kallikrein 694:703 rat kallikrein 690:703 rat kallikrein 690:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 13 theme primary 572:578 arg1 structure 580:588 the primary structure 568:588 the primary structure of human urinary kallikrein 568:616 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 1 14 theme amino 85:89 arg1 sequence 96:103 The amino acid sequence 81:103 The amino acid sequence of human urinary kallikrein 81:131 The amino acid sequence of human urinary kallikrein was determined by protein-chemical methods. 3163150 4 15 dep three 334:338 arg1 linked 369:374 linked 369:374 are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) 348:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 0 16 theme urinary 6:12 arg1 sequence 37:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence and carbohydrate attachment sites. 3163150 5 17 theme serine 637:642 arg1 proteinases 644:654 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 17 theme serine 637:642 arg1 kallikrein 675:684 porcine kallikrein 667:684 porcine kallikrein 667:684 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 17 theme serine 637:642 arg1 kallikrein 694:703 rat kallikrein 690:703 rat kallikrein 690:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 18 theme structure 580:588 arg1 Comparison 554:563 Comparison 554:563 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein 554:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 1 19 theme acid 91:94 arg1 sequence 96:103 The amino acid sequence 81:103 The amino acid sequence of human urinary kallikrein 81:131 The amino acid sequence of human urinary kallikrein was determined by protein-chemical methods. 3163150 0 20 theme Human 0:4 arg1 sequence 37:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence and carbohydrate attachment sites. 3163150 0 21 theme carbohydrate 50:61 arg1 sites 74:78 carbohydrate attachment sites 50:78 carbohydrate attachment sites 50:78 Human urinary kallikrein--amino acid sequence and carbohydrate attachment sites. 3163150 2 22 contain contains 195:202 arg1 chain 189:193 The peptide chain 177:193 The peptide chain 177:193 The peptide chain contains 238 amino acid residues. 3163150 2 22 contain contains 195:202 arg2 residues 219:226 238 amino acid residues 204:226 238 amino acid residues 204:226 The peptide chain contains 238 amino acid residues. 3163150 0 23 theme acid 32:35 arg1 sequence 37:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence and carbohydrate attachment sites. 3163150 5 24 theme high 713:716 arg1 degree 718:723 a high degree 711:723 a high degree of homology 711:735 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 25 theme human 593:597 arg1 kallikrein 607:616 human urinary kallikrein 593:616 human urinary kallikrein 593:616 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 0 26 theme kallikrein--amino 14:30 arg1 sequence 37:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence 0:44 Human urinary kallikrein--amino acid sequence and carbohydrate attachment sites. 3163150 1 27 theme human 108:112 arg1 kallikrein 122:131 human urinary kallikrein 108:131 human urinary kallikrein 108:131 The amino acid sequence of human urinary kallikrein was determined by protein-chemical methods. 3163150 4 28 theme side 301:304 arg1 chains 306:311 Six carbohydrate side chains 284:311 Six carbohydrate side chains 284:311 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 29 theme carbohydrate 288:299 arg1 chains 306:311 Six carbohydrate side chains 284:311 Six carbohydrate side chains 284:311 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 5 30 dep proteinases 644:654 arg1 proteinases 644:654 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 30 dep proteinases 644:654 arg1 kallikrein 675:684 porcine kallikrein 667:684 porcine kallikrein 667:684 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 30 dep proteinases 644:654 arg1 kallikrein 694:703 rat kallikrein 690:703 rat kallikrein 690:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 31 theme homology 728:735 arg1 degree 718:723 a high degree 711:723 a high degree of homology 711:735 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 2 32 theme peptide 181:187 arg1 chain 189:193 The peptide chain 177:193 The peptide chain 177:193 The peptide chain contains 238 amino acid residues. 3163150 0 33 theme attachment 63:72 arg1 sites 74:78 carbohydrate attachment sites 50:78 carbohydrate attachment sites 50:78 Human urinary kallikrein--amino acid sequence and carbohydrate attachment sites. 3163150 4 34 theme threonine 521:529 arg1 residue 531:537 one threonine residue 517:537 one threonine residue 517:537 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 5 35 theme rat 690:692 arg1 proteinases 644:654 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 other serine proteinases especially porcine kallikrein and rat kallikrein 631:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 5 35 theme rat 690:692 arg1 kallikrein 694:703 rat kallikrein 690:703 rat kallikrein 690:703 Comparison of the primary structure of human urinary kallikrein with that of other serine proteinases especially porcine kallikrein and rat kallikrein shows a high degree of homology. 3163150 1 36 theme urinary 114:120 arg1 kallikrein 122:131 human urinary kallikrein 108:131 human urinary kallikrein 108:131 The amino acid sequence of human urinary kallikrein was determined by protein-chemical methods. 3163150 4 37 located present 399:405 arg2 asparagine 379:388 asparagine 379:388 asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) 379:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 37 located present 399:405 arg1 Asn-Xaa-Thr 430:440 the acceptor sequon Asn-Xaa-Thr 410:440 the acceptor sequon Asn-Xaa-Thr (or -Ser) 410:450 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 37 located present 399:405 arg1 -Ser 446:449 -Ser 446:449 -Ser 446:449 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 1 38 theme kallikrein 122:131 arg1 sequence 96:103 The amino acid sequence 81:103 The amino acid sequence of human urinary kallikrein 81:131 The amino acid sequence of human urinary kallikrein was determined by protein-chemical methods. 3163150 2 39 theme acid 214:217 arg1 residues 219:226 238 amino acid residues 204:226 238 amino acid residues 204:226 The peptide chain contains 238 amino acid residues. 3163150 6 40 theme IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA 738:776 arg1 AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN 778:816 IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN 738:816 IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN 738:816 IVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQWVLTA AHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFN MSLLENHTRQADEDYSHDLMLLRLTEPADTITDAVKVVE LPTQEPEVGSTCLASGWGSIEPENFSFPDDLQCVDLKIL PNDECEKAHVQKVTDFMLCVGHLEGGKDTCVGDSGGPLM CDGVLQGVTSWGYVPCGTPNKPSVAVRVLSYVKWIEDTIAENS. 3163150 4 41 from Asn-Xaa-Thr 430:440 arg1 present 399:405 present 399:405 present 399:405 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 2 42 theme amino 208:212 arg1 residues 219:226 238 amino acid residues 204:226 238 amino acid residues 204:226 The peptide chain contains 238 amino acid residues. 3163150 1 43 theme protein-chemical 151:166 arg1 methods 168:174 protein-chemical methods 151:174 protein-chemical methods 151:174 The amino acid sequence of human urinary kallikrein was determined by protein-chemical methods. 3163150 4 44 theme them 343:346 arg1 them 343:346 them 343:346 them 343:346 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 44 theme them 343:346 arg1 three 334:338 three 334:338 three 334:338 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 45 attach linked 483:488 arg1 residues 504:511 two serine residues 493:511 two serine residues 493:511 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 45 attach linked 483:488 arg2 three 456:460 three 456:460 three 456:460 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 45 attach linked 483:488 arg2 them 343:346 them 343:346 them 343:346 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 45 attach linked 483:488 arg1 residue 531:537 one threonine residue 517:537 one threonine residue 517:537 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 3163150 4 45 attach linked 483:488 arg2 three 334:338 three 334:338 three 334:338 Six carbohydrate side chains could be identified, three of them are N-glycosidically linked to asparagine which is present in the acceptor sequon Asn-Xaa-Thr (or -Ser) and three are O-glycosidically linked to two serine residues and one threonine residue, respectively. 11325591 0 0 theme key 75:77 arg1 caspase-3 64:72 caspase-3 64:72 caspase-3 64:72 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 0 0 theme key 75:77 arg1 mediators 79:87 key mediators 75:87 key mediators of cell death with cleavage specificity for aspartic acid in P1 75:151 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 1 1 theme cysteine 282:289 arg1 family 313:318 the caspase (cysteine aspartyl proteinases) family 269:318 the caspase (cysteine aspartyl proteinases) family 269:318 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 1 theme cysteine 282:289 arg1 proteinases 300:310 cysteine aspartyl proteinases 282:310 cysteine aspartyl proteinases 282:310 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 2 2 theme granzyme 546:553 arg1 B 555:555 the human granzyme B 536:555 the human granzyme B 536:555 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 2 3 theme three-dimensional 490:506 arg1 structure 508:516 the three-dimensional structure 486:516 the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor 486:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 2 4 theme complex 525:531 arg1 structure 508:516 the three-dimensional structure 486:516 the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor 486:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 1 5 theme cytotoxic 215:223 arg1 T-lymphocyte 225:236 cytotoxic T-lymphocyte 215:236 cytotoxic T-lymphocyte (CTL) granules 215:251 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 5 theme cytotoxic 215:223 arg1 CTL 239:241 CTL 239:241 CTL 239:241 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 6 from acid 368:371 arg1 P1 376:377 P1 376:377 P1 376:377 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 7 theme aspartyl 291:298 arg1 family 313:318 the caspase (cysteine aspartyl proteinases) family 269:318 the caspase (cysteine aspartyl proteinases) family 269:318 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 7 theme aspartyl 291:298 arg1 proteinases 300:310 cysteine aspartyl proteinases 282:310 cysteine aspartyl proteinases 282:310 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 2 8 theme human 540:544 arg1 B 555:555 the human granzyme B 536:555 the human granzyme B 536:555 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 2 9 with complex 525:531 arg1 inhibitor 593:601 a potent tetrapeptide aldehyde inhibitor 562:601 a potent tetrapeptide aldehyde inhibitor 562:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 1 10 theme T-lymphocyte 225:236 arg1 granules 244:251 cytotoxic T-lymphocyte (CTL) granules 215:251 cytotoxic T-lymphocyte (CTL) granules 215:251 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 11 dep proteins 1078:1085 arg1 these 1072:1076 these 1072:1076 these 1072:1076 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 12 theme human 635:639 arg1 B 650:650 human granzyme B 635:650 human granzyme B 635:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 13 from site 729:732 arg1 caspases 761:768 the apoptosis-promoting caspases 737:768 the apoptosis-promoting caspases 737:768 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 2 14 theme aldehyde 584:591 arg1 inhibitor 593:601 a potent tetrapeptide aldehyde inhibitor 562:601 a potent tetrapeptide aldehyde inhibitor 562:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 0 15 from acid 142:145 arg1 P1 150:151 P1 150:151 P1 150:151 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 1 16 theme critical 388:395 arg1 roles 397:401 critical roles 388:401 critical roles 388:401 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 17 theme above 853:857 arg1 differences 859:869 The above differences 849:869 The above differences 849:869 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 0 18 theme death 97:101 arg1 caspase-3 64:72 caspase-3 64:72 caspase-3 64:72 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 0 18 theme death 97:101 arg1 mediators 79:87 key mediators 75:87 key mediators of cell death with cleavage specificity for aspartic acid in P1 75:151 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 1 19 theme family 313:318 arg1 B 174:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 19 theme family 313:318 arg1 members 258:264 members 258:264 members of the caspase (cysteine aspartyl proteinases) family 258:318 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 2 20 theme potent 564:569 arg1 inhibitor 593:601 a potent tetrapeptide aldehyde inhibitor 562:601 a potent tetrapeptide aldehyde inhibitor 562:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 2 21 theme tetrapeptide 571:582 arg1 inhibitor 593:601 a potent tetrapeptide aldehyde inhibitor 562:601 a potent tetrapeptide aldehyde inhibitor 562:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 0 22 theme cell 92:95 arg1 death 97:101 cell death 92:101 cell death 92:101 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 1 23 theme BACKGROUND 154:163 arg1 B 174:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 0 24 theme three-dimensional 4:20 arg1 structure 22:30 The three-dimensional structure 0:30 The three-dimensional structure of human granzyme B 0:50 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 3 25 dep granzyme 825:832 arg1 B. 834:835 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 25 dep granzyme 825:832 arg1 CONCLUSIONS 837:847 CONCLUSIONS 837:847 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 26 theme disease 1095:1101 arg1 states 1103:1108 the disease states 1091:1108 the disease states 1091:1108 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 27 theme Granzyme 165:172 arg1 B 174:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 2 28 theme B 555:555 arg1 complex 525:531 the complex 521:531 the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor 521:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 1 29 theme biochemical 410:420 arg1 events 422:427 the biochemical events 406:427 the biochemical events that culminate in cell death 406:456 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 30 theme S1 621:622 arg1 B 650:650 human granzyme B 635:650 human granzyme B 635:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 30 theme S1 621:622 arg1 larger 669:674 larger 669:674 larger 669:674 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 30 theme S1 621:622 arg1 subsite 624:630 The Asp-specific S1 subsite 604:630 The Asp-specific S1 subsite of human granzyme B 604:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 31 theme inhibitors 1015:1024 arg1 design 996:1001 the design 992:1001 the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 992:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 0 32 theme cleavage 108:115 arg1 specificity 117:127 cleavage specificity 108:127 cleavage specificity for aspartic acid in P1 108:151 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 0 33 with mediators 79:87 arg1 specificity 117:127 cleavage specificity 108:127 cleavage specificity for aspartic acid in P1 108:151 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 3 34 from subsite 810:816 arg1 granzyme 825:832 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 35 dep CONCLUSIONS 837:847 arg1 account 871:877 account 871:877 account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases 871:978 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 35 dep CONCLUSIONS 837:847 arg1 enable 985:990 enable 985:990 enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 985:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 36 theme unique 327:332 arg1 specificity 343:353 a unique cleavage specificity 325:353 a unique cleavage specificity for aspartic acid in P1 325:377 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 0 37 theme granzyme 41:48 arg1 B 50:50 human granzyme B 35:50 human granzyme B 35:50 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 3 38 theme physiological 1045:1057 arg1 functions 1059:1067 the physiological functions 1041:1067 the physiological functions of these proteins and the disease states with which they are associated 1041:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 39 theme cleavage 334:341 arg1 specificity 343:353 a unique cleavage specificity 325:353 a unique cleavage specificity for aspartic acid in P1 325:377 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 0 40 theme human 35:39 arg1 B 50:50 human granzyme B 35:50 human granzyme B 35:50 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 3 41 theme specific 1006:1013 arg1 inhibitors 1015:1024 specific inhibitors 1006:1024 specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 1006:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 2 42 dep RESULTS 459:465 arg1 determined 475:484 determined 475:484 have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor 470:601 RESULTS We have determined the three-dimensional structure of the complex of the human granzyme B with a potent tetrapeptide aldehyde inhibitor. 11325591 3 43 theme Asp-specific 716:727 arg1 site 729:732 the corresponding Asp-specific site 698:732 the corresponding Asp-specific site in the apoptosis-promoting caspases 698:768 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 44 from granzymes 202:210 arg1 granules 244:251 cytotoxic T-lymphocyte (CTL) granules 215:251 cytotoxic T-lymphocyte (CTL) granules 215:251 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 45 from variation 887:895 arg1 specificity 910:920 substrate specificity 900:920 substrate specificity 900:920 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 46 theme granzyme 928:935 arg1 proteases 953:961 other serine proteases 940:961 other serine proteases 940:961 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 46 theme granzyme 928:935 arg1 caspases 971:978 the caspases 967:978 the caspases 967:978 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 46 theme granzyme 928:935 arg1 B 937:937 granzyme B 928:937 granzyme B 928:937 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 0 47 theme B 50:50 arg1 structure 22:30 The three-dimensional structure 0:30 The three-dimensional structure of human granzyme B 0:50 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 3 48 theme corresponding 702:714 arg1 site 729:732 the corresponding Asp-specific site 698:732 the corresponding Asp-specific site in the apoptosis-promoting caspases 698:768 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 0 49 theme aspartic 133:140 arg1 acid 142:145 aspartic acid 133:145 aspartic acid in P1 133:151 The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. 11325591 1 50 theme aspartic 359:366 arg1 acid 368:371 aspartic acid 359:371 aspartic acid in P1 359:377 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 51 theme substrate 900:908 arg1 specificity 910:920 substrate specificity 900:920 substrate specificity 900:920 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 52 theme caspase 273:279 arg1 family 313:318 the caspase (cysteine aspartyl proteinases) family 269:318 the caspase (cysteine aspartyl proteinases) family 269:318 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 52 theme caspase 273:279 arg1 proteinases 300:310 cysteine aspartyl proteinases 282:310 cysteine aspartyl proteinases 282:310 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 53 theme proteins 1078:1085 arg1 functions 1059:1067 the physiological functions 1041:1067 the physiological functions of these proteins and the disease states with which they are associated 1041:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 54 theme cell 447:450 arg1 death 452:456 cell death 447:456 cell death 447:456 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 55 theme rat 821:823 arg1 granzyme 825:832 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 821:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 56 theme other 940:944 arg1 proteases 953:961 other serine proteases 940:961 other serine proteases 940:961 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 56 theme other 940:944 arg1 B 937:937 granzyme B 928:937 granzyme B 928:937 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 57 theme abundant 193:200 arg1 granzymes 202:210 the most abundant granzymes 184:210 the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules 184:251 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 58 theme Asp-specific 608:619 arg1 B 650:650 human granzyme B 635:650 human granzyme B 635:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 58 theme Asp-specific 608:619 arg1 larger 669:674 larger 669:674 larger 669:674 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 58 theme Asp-specific 608:619 arg1 subsite 624:630 The Asp-specific S1 subsite 604:630 The Asp-specific S1 subsite of human granzyme B 604:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 59 theme serine 946:951 arg1 proteases 953:961 other serine proteases 940:961 other serine proteases 940:961 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 59 theme serine 946:951 arg1 B 937:937 granzyme B 928:937 granzyme B 928:937 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 60 theme states 1103:1108 arg1 functions 1059:1067 the physiological functions 1041:1067 the physiological functions of these proteins and the disease states with which they are associated 1041:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 61 theme corresponding 796:808 arg1 subsite 810:816 the corresponding subsite 792:816 the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated 792:1139 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 62 theme granzymes 202:210 arg1 granzymes 202:210 the most abundant granzymes 184:210 the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules 184:251 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 62 theme granzymes 202:210 arg1 one 177:179 one 177:179 one 177:179 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 63 theme granzyme 641:648 arg1 B 650:650 human granzyme B 635:650 human granzyme B 635:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 64 theme B 650:650 arg1 B 650:650 human granzyme B 635:650 human granzyme B 635:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 64 theme B 650:650 arg1 larger 669:674 larger 669:674 larger 669:674 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 3 64 theme B 650:650 arg1 subsite 624:630 The Asp-specific S1 subsite 604:630 The Asp-specific S1 subsite of human granzyme B 604:650 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 11325591 1 65 contain have 320:323 arg1 granzymes 202:210 the most abundant granzymes 184:210 the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules 184:251 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 65 contain have 320:323 arg1 one 177:179 one 177:179 one 177:179 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 65 contain have 320:323 arg1 B 174:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B 154:174 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 65 contain have 320:323 arg1 members 258:264 members 258:264 members of the caspase (cysteine aspartyl proteinases) family 258:318 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 1 65 contain have 320:323 arg2 specificity 343:353 a unique cleavage specificity 325:353 a unique cleavage specificity for aspartic acid in P1 325:377 BACKGROUND Granzyme B, one of the most abundant granzymes in cytotoxic T-lymphocyte (CTL) granules, and members of the caspase (cysteine aspartyl proteinases) family have a unique cleavage specificity for aspartic acid in P1 and play critical roles in the biochemical events that culminate in cell death. 11325591 3 66 theme apoptosis-promoting 741:759 arg1 caspases 761:768 the apoptosis-promoting caspases 737:768 the apoptosis-promoting caspases 737:768 The Asp-specific S1 subsite of human granzyme B is significantly larger and less charged than the corresponding Asp-specific site in the apoptosis-promoting caspases, and also larger than the corresponding subsite in rat granzyme B. CONCLUSIONS The above differences account for the variation in substrate specificity among granzyme B, other serine proteases and the caspases, and enable the design of specific inhibitors that can probe the physiological functions of these proteins and the disease states with which they are associated. 30356213 4 0 theme protein 749:755 arg1 inability 698:706 an inability 695:706 an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate 695:808 Precise mechanistic studies have so far been prevented by an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate. 30356213 2 1 theme mouse 355:359 arg1 knockout5 361:369 mouse knockout5 355:369 mouse knockout5 355:369 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 5 2 theme enzyme 961:966 arg1 process 987:993 a palladium-mediated enzyme activation 'shunt' process 940:993 a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation 940:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 1 3 from molecule 160:167 arg1 animals 180:186 animals 180:186 animals 180:186 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 1 3 from molecule 160:167 arg1 humans 172:177 humans 172:177 humans 172:177 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 1 3 from molecule 160:167 arg1 fungi1 192:197 fungi1 192:197 fungi1 192:197 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 6 4 theme distinct 1105:1112 arg1 stages 1114:1119 distinct stages 1105:1119 distinct stages 1105:1119 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 8 5 from tool 1517:1520 arg1 study 1529:1533 the study 1525:1533 the study of mechanism 1525:1546 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 4 6 gly glycoforms 730:739 arg1 access 711:716 access homogeneous glycoforms 711:739 access homogeneous glycoforms 711:739 Precise mechanistic studies have so far been prevented by an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate. 30356213 8 7 theme active 1433:1438 arg1 enzymes 1440:1446 active enzymes 1433:1446 active enzymes 1433:1446 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 5 8 theme activation 968:977 arg1 process 987:993 a palladium-mediated enzyme activation 'shunt' process 940:993 a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation 940:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 2 9 theme glycogen 282:289 arg1 formation 291:299 glycogen formation 282:299 glycogen formation 282:299 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 7 10 theme proof-read 1263:1272 arg1 mechanism 1275:1283 a tolerant but 'proof-read' mechanism 1247:1283 a tolerant but 'proof-read' mechanism that underlies the precision of this metabolic process 1247:1338 This reveals a tolerant but 'proof-read' mechanism that underlies the precision of this metabolic process. 30356213 8 11 theme mechanism 1538:1546 arg1 study 1529:1533 the study 1525:1533 the study of mechanism 1525:1546 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 3 12 theme own 523:525 arg1 autoglucosylation 536:552 its own stepwise autoglucosylation 519:552 its own stepwise autoglucosylation 519:552 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 7 13 theme tolerant 1249:1256 arg1 mechanism 1275:1283 a tolerant but 'proof-read' mechanism 1247:1283 a tolerant but 'proof-read' mechanism that underlies the precision of this metabolic process 1247:1338 This reveals a tolerant but 'proof-read' mechanism that underlies the precision of this metabolic process. 30356213 1 14 dep energy 144:149 arg1 hence 138:142 hence 138:142 hence 138:142 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 4 15 theme access 711:716 arg1 glycoforms 730:739 access homogeneous glycoforms 711:739 access homogeneous glycoforms 711:739 Precise mechanistic studies have so far been prevented by an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate. 30356213 6 16 theme GYG 1051:1053 arg1 intermediates 1055:1067 GYG intermediates 1051:1067 GYG intermediates 1051:1067 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 5 17 theme shunt 980:984 arg1 process 987:993 a palladium-mediated enzyme activation 'shunt' process 940:993 a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation 940:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 5 18 theme different 860:868 arg1 states 898:903 different, homogeneously glucosylated states 860:903 different, homogeneously glucosylated states of GYG 860:910 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 7 19 theme process 1332:1338 arg1 precision 1304:1312 the precision 1300:1312 the precision of this metabolic process 1300:1338 This reveals a tolerant but 'proof-read' mechanism that underlies the precision of this metabolic process. 30356213 6 20 theme triphasic 1156:1164 arg1 kinetics 1166:1173 triphasic kinetics 1156:1173 triphasic kinetics 1156:1173 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 1 21 theme glycogen 101:108 arg1 Biosynthesis 85:96 Biosynthesis 85:96 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, 85:198 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 5 22 dep different 860:868 arg1 glucosylated 885:896 glucosylated 885:896 glucosylated 885:896 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 0 23 theme enzyme 19:24 arg1 activation 26:35 Palladium-mediated enzyme activation 0:35 Palladium-mediated enzyme activation 0:35 Palladium-mediated enzyme activation suggests multiphase initiation of glycogenesis. 30356213 8 24 dep direct 1370:1375 arg1 controlled 1389:1398 controlled 1389:1398 controlled 1389:1398 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 2 25 theme neurodegenerative 307:323 arg1 disease2-4 339:348 neurodegenerative and metabolic disease2-4 307:348 neurodegenerative and metabolic disease2-4 307:348 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 6 26 theme catalytic 1083:1091 arg1 activity 1093:1100 catalytic activity 1083:1100 catalytic activity 1083:1100 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 2 27 theme human 385:389 arg1 mutations6 391:400 inherited human mutations6 375:400 inherited human mutations6 375:400 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 0 28 theme Palladium-mediated 0:17 arg1 activation 26:35 Palladium-mediated enzyme activation 0:35 Palladium-mediated enzyme activation 0:35 Palladium-mediated enzyme activation suggests multiphase initiation of glycogenesis. 30356213 6 29 theme kinetics 1166:1173 arg1 plasticity 1189:1198 substrate plasticity 1179:1198 substrate plasticity 1179:1198 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 6 29 theme kinetics 1166:1173 arg1 discovery 1143:1151 discovery 1143:1151 discovery of triphasic kinetics 1143:1173 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 2 30 theme inherited 375:383 arg1 mutations6 391:400 inherited human mutations6 375:400 inherited human mutations6 375:400 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 8 31 theme intermediate 1410:1421 arg1 states 1423:1428 intermediate states 1410:1428 intermediate states of active enzymes 1410:1446 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 1 32 theme essential 115:123 arg1 energy 144:149 energy 144:149 energy 144:149 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 1 32 theme essential 115:123 arg1 glucose 125:131 essential glucose 115:131 the essential glucose (and hence energy) storage molecule in humans, animals and fungi1 111:197 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 5 33 theme bond 1016:1019 arg1 formation 1021:1029 on-protein C-C bond formation 1001:1029 on-protein C-C bond formation 1001:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 3 34 theme seed 451:454 arg1 GYG 436:438 GYG 436:438 GYG 436:438 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 3 34 theme seed 451:454 arg1 core 456:459 a 'seed core' 448:460 a 'seed core' for the formation of the glycogen particle 448:503 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 8 35 theme direct 1370:1375 arg1 access 1400:1405 direct, chemically controlled access 1370:1405 direct, chemically controlled access to intermediate states of active enzymes 1370:1446 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 8 36 theme such 1462:1465 arg1 tool 1517:1520 a powerful tool 1506:1520 a powerful tool 1506:1520 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 8 36 theme such 1462:1465 arg1 activation 1486:1495 such ligation-dependent activation 1462:1495 such ligation-dependent activation 1462:1495 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 1 37 theme glucose 125:131 arg1 glycogen 101:108 glycogen 101:108 glycogen 101:108 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 1 37 theme glucose 125:131 arg1 molecule 160:167 the essential glucose (and hence energy) storage molecule 111:167 the essential glucose (and hence energy) storage molecule in humans, animals and fungi1 111:197 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 2 38 theme glycogen 416:423 arg1 synthesis 425:433 glycogen synthesis 416:433 glycogen synthesis 416:433 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 5 39 theme unprecedented 829:841 arg1 access 850:855 unprecedented direct access 829:855 unprecedented direct access to different, homogeneously glucosylated states of GYG 829:910 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 3 40 theme bound 575:579 arg1 chain 603:607 a covalently bound gluco-oligosaccharide chain 562:607 a covalently bound gluco-oligosaccharide chain 562:607 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 7 41 theme metabolic 1322:1330 arg1 process 1332:1338 this metabolic process 1317:1338 this metabolic process 1317:1338 This reveals a tolerant but 'proof-read' mechanism that underlies the precision of this metabolic process. 30356213 4 42 theme mechanistic 645:655 arg1 studies 657:663 Precise mechanistic studies 637:663 Precise mechanistic studies 637:663 Precise mechanistic studies have so far been prevented by an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate. 30356213 0 43 theme multiphase 46:55 arg1 initiation 57:66 multiphase initiation 46:66 multiphase initiation of glycogenesis 46:82 Palladium-mediated enzyme activation suggests multiphase initiation of glycogenesis. 30356213 3 44 theme gluco-oligosaccharide 581:601 arg1 chain 603:607 a covalently bound gluco-oligosaccharide chain 562:607 a covalently bound gluco-oligosaccharide chain 562:607 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 5 45 theme direct 843:848 arg1 access 850:855 unprecedented direct access 829:855 unprecedented direct access to different, homogeneously glucosylated states of GYG 829:910 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 2 46 theme GYG 405:407 arg1 mutations6 391:400 inherited human mutations6 375:400 inherited human mutations6 375:400 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 2 46 theme GYG 405:407 arg1 knockout5 361:369 mouse knockout5 355:369 mouse knockout5 355:369 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 0 47 theme glycogenesis 71:82 arg1 initiation 57:66 multiphase initiation 46:66 multiphase initiation of glycogenesis 46:82 Palladium-mediated enzyme activation suggests multiphase initiation of glycogenesis. 30356213 8 48 theme ligation-dependent 1467:1484 arg1 tool 1517:1520 a powerful tool 1506:1520 a powerful tool 1506:1520 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 8 48 theme ligation-dependent 1467:1484 arg1 activation 1486:1495 such ligation-dependent activation 1462:1495 such ligation-dependent activation 1462:1495 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 5 49 theme GYG 908:910 arg1 states 898:903 different, homogeneously glucosylated states 860:903 different, homogeneously glucosylated states of GYG 860:910 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 5 50 theme palladium-mediated 942:959 arg1 process 987:993 a palladium-mediated enzyme activation 'shunt' process 940:993 a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation 940:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 6 51 theme substrates 1222:1231 arg1 use 1209:1211 GYG's use 1203:1211 GYG's use of sugar substrates 1203:1231 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 8 52 theme enzymes 1440:1446 arg1 states 1423:1428 intermediate states 1410:1428 intermediate states of active enzymes 1410:1446 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 6 53 theme sugar 1216:1220 arg1 substrates 1222:1231 sugar substrates 1216:1231 sugar substrates 1216:1231 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 3 54 theme initiation 612:621 arg1 site 623:626 initiation site Tyr 195 612:634 initiation site Tyr 195 612:634 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 8 55 theme access 1400:1405 arg1 demonstration 1353:1365 The present demonstration 1341:1365 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes 1341:1446 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 2 56 theme metabolic 329:337 arg1 disease2-4 339:348 neurodegenerative and metabolic disease2-4 307:348 neurodegenerative and metabolic disease2-4 307:348 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 3 57 theme stepwise 527:534 arg1 autoglucosylation 536:552 its own stepwise autoglucosylation 519:552 its own stepwise autoglucosylation 519:552 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 6 58 theme Careful 1032:1038 arg1 mimicry 1040:1046 Careful mimicry 1032:1046 Careful mimicry of GYG intermediates 1032:1067 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 6 59 theme substrate 1179:1187 arg1 plasticity 1189:1198 substrate plasticity 1179:1198 substrate plasticity 1179:1198 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 8 60 theme powerful 1508:1515 arg1 tool 1517:1520 a powerful tool 1506:1520 a powerful tool 1506:1520 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 8 60 theme powerful 1508:1515 arg1 activation 1486:1495 such ligation-dependent activation 1462:1495 such ligation-dependent activation 1462:1495 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 8 61 theme present 1345:1351 arg1 demonstration 1353:1365 The present demonstration 1341:1365 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes 1341:1446 The present demonstration of direct, chemically controlled access to intermediate states of active enzymes suggests that such ligation-dependent activation could be a powerful tool in the study of mechanism. 30356213 3 62 theme particle 496:503 arg1 formation 470:478 the formation 466:478 the formation of the glycogen particle 466:503 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 4 63 theme homogeneous 718:728 arg1 glycoforms 730:739 access homogeneous glycoforms 711:739 access homogeneous glycoforms 711:739 Precise mechanistic studies have so far been prevented by an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate. 30356213 4 64 theme Precise 637:643 arg1 studies 657:663 Precise mechanistic studies 637:663 Precise mechanistic studies 637:663 Precise mechanistic studies have so far been prevented by an inability to access homogeneous glycoforms of this protein, which unusually acts as both catalyst and substrate. 30356213 5 65 theme on-protein 1001:1010 arg1 formation 1021:1029 on-protein C-C bond formation 1001:1029 on-protein C-C bond formation 1001:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 6 66 theme intermediates 1055:1067 arg1 mimicry 1040:1046 Careful mimicry 1032:1046 Careful mimicry of GYG intermediates 1032:1067 Careful mimicry of GYG intermediates recapitulates catalytic activity at distinct stages, which in turn allows discovery of triphasic kinetics and substrate plasticity in GYG's use of sugar substrates. 30356213 3 67 theme glycogen 487:494 arg1 particle 496:503 the glycogen particle 483:503 the glycogen particle 483:503 GYG acts as a 'seed core' for the formation of the glycogen particle by catalysing its own stepwise autoglucosylation to form a covalently bound gluco-oligosaccharide chain at initiation site Tyr 195. 30356213 2 68 from Deficiencies 266:277 arg1 formation 291:299 glycogen formation 282:299 glycogen formation 282:299 Deficiencies in glycogen formation cause neurodegenerative and metabolic disease2-4, and mouse knockout5 and inherited human mutations6 of GYG impair glycogen synthesis. 30356213 1 69 theme glycosyltransferase 220:238 arg1 glycogenin 248:257 glycogenin 248:257 glycogenin (GYG) 248:263 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 1 69 theme glycosyltransferase 220:238 arg1 enzyme 240:245 the glycosyltransferase enzyme 216:245 the glycosyltransferase enzyme 216:245 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 5 70 theme C-C 1012:1014 arg1 formation 1021:1029 on-protein C-C bond formation 1001:1029 on-protein C-C bond formation 1001:1029 Here we show that unprecedented direct access to different, homogeneously glucosylated states of GYG can be accomplished through a palladium-mediated enzyme activation 'shunt' process using on-protein C-C bond formation. 30356213 1 71 theme storage 152:158 arg1 glycogen 101:108 glycogen 101:108 glycogen 101:108 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 30356213 1 71 theme storage 152:158 arg1 molecule 160:167 the essential glucose (and hence energy) storage molecule 111:167 the essential glucose (and hence energy) storage molecule in humans, animals and fungi1 111:197 Biosynthesis of glycogen, the essential glucose (and hence energy) storage molecule in humans, animals and fungi1, is initiated by the glycosyltransferase enzyme, glycogenin (GYG). 22160680 6 0 theme conformational 1043:1056 arg1 plasticity 1058:1067 the conformational plasticity 1039:1067 the conformational plasticity of glycogenin and coexistence of two modes of glucosylation 1039:1127 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 6 1 theme coexistence 1087:1097 arg1 plasticity 1058:1067 the conformational plasticity 1039:1067 the conformational plasticity of glycogenin and coexistence of two modes of glucosylation 1039:1127 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 5 2 theme storage 925:931 arg1 disease 933:939 glycogen storage disease XV 916:942 glycogen storage disease XV 916:942 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 2 3 theme conformational 382:395 arg1 switch 397:402 a dynamic conformational switch 372:402 a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose 372:475 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 4 4 from site 722:725 arg1 mode 763:766 either an intra- or intersubunit mode 730:766 mode 763:766 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 3 5 theme segment 612:618 arg1 movement 566:573 major movement 560:573 major movement of an approximately 30-residue "lid" segment covering the active site 560:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 3 5 theme segment 612:618 arg1 Tyr195 548:553 Tyr195 548:553 Tyr195 548:553 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 2 6 theme glycogenin 324:333 arg1 snapshots 305:313 crystallographic snapshots 288:313 crystallographic snapshots of human glycogenin 288:333 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 1 7 attach attached 175:182 arg3 Tyr195 197:202 Tyr195 197:202 Tyr195 197:202 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 1 7 attach attached 175:182 arg2 chain 158:162 a maltosaccharide chain 140:162 a maltosaccharide chain covalently attached to itself on Tyr195 140:202 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 1 7 attach attached 175:182 arg1 itself 187:192 itself 187:192 itself 187:192 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 2 8 theme human 318:322 arg1 glycogenin 324:333 human glycogenin 318:333 human glycogenin 318:333 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 3 9 theme polypeptide 517:527 arg1 stretch 529:535 a polypeptide stretch 515:535 a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site 515:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 3 10 theme stretch 529:535 arg1 ordering 503:510 the ordering 499:510 the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site 499:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 2 11 theme sugar 453:457 arg1 donor 459:463 the sugar donor UDP-glucose 449:475 the sugar donor UDP-glucose 449:475 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 6 12 theme modes 1106:1110 arg1 coexistence 1087:1097 coexistence 1087:1097 coexistence 1087:1097 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 6 12 theme modes 1106:1110 arg1 glycogenin 1072:1081 glycogenin 1072:1081 glycogenin 1072:1081 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 4 13 dep donor 833:837 arg1 groups 858:863 sugar groups 852:863 sugar groups 852:863 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 3 14 contain containing 537:546 arg1 stretch 529:535 a polypeptide stretch 515:535 a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site 515:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 3 14 contain containing 537:546 arg2 movement 566:573 major movement 560:573 major movement of an approximately 30-residue "lid" segment covering the active site 560:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 3 14 contain containing 537:546 arg2 Tyr195 548:553 Tyr195 548:553 Tyr195 548:553 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 4 15 theme steric 800:805 arg1 factors 807:813 steric factors 800:813 steric factors 800:813 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 4 16 theme maltosaccharide 684:698 arg1 chain 700:704 the nascent maltosaccharide chain 672:704 the nascent maltosaccharide chain 672:704 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 6 17 theme glucosylation 1115:1127 arg1 modes 1106:1110 two modes 1102:1110 two modes of glucosylation 1102:1127 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 4 18 theme dependent 768:776 arg1 mode 763:766 either an intra- or intersubunit mode 730:766 mode 763:766 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 2 19 theme active 423:428 arg1 states 430:435 ground and active states 412:435 ground and active states mediated by the sugar donor UDP-glucose 412:475 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 0 20 theme Conformational 0:13 arg1 plasticity 15:24 Conformational plasticity 0:24 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis 0:99 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis. 22160680 2 21 theme reaction 346:353 arg1 cycle 355:359 its reaction cycle 342:359 its reaction cycle 342:359 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 0 22 theme glycogenin 29:38 arg1 plasticity 15:24 Conformational plasticity 0:24 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis 0:99 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis. 22160680 4 23 theme chain 783:787 arg1 length 789:794 chain length 783:794 chain length 783:794 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 1 24 theme stepwise 210:217 arg1 reaction 233:240 a stepwise glucosylation reaction 208:240 a stepwise glucosylation reaction 208:240 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 3 25 theme " 610:610 arg1 segment 612:618 an approximately 30-residue "lid" segment 578:618 an approximately 30-residue "lid" segment covering the active site 578:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 5 26 theme ground 979:984 arg1 state 986:990 the ground state 975:990 the ground state 975:990 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 1 27 theme glucosylation 219:231 arg1 reaction 233:240 a stepwise glucosylation reaction 208:240 a stepwise glucosylation reaction 208:240 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 5 28 from state 986:990 arg1 mutation 893:900 The Thr83Met mutation 880:900 The Thr83Met mutation 880:900 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 5 28 from state 986:990 arg1 locked 965:970 locked 965:970 locked 965:970 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 4 29 theme nascent 676:682 arg1 chain 700:704 the nascent maltosaccharide chain 672:704 the nascent maltosaccharide chain 672:704 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 5 30 theme Thr83Met 884:891 arg1 mutation 893:900 The Thr83Met mutation 880:900 The Thr83Met mutation 880:900 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 5 30 theme Thr83Met 884:891 arg1 locked 965:970 locked 965:970 locked 965:970 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 3 31 theme active 633:638 arg1 site 640:643 the active site 629:643 the active site 629:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 22160680 6 32 theme glycogenin 1072:1081 arg1 plasticity 1058:1067 the conformational plasticity 1039:1067 the conformational plasticity of glycogenin and coexistence of two modes of glucosylation 1039:1127 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 0 33 theme substrate 64:72 arg1 plasticity 15:24 Conformational plasticity 0:24 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis 0:99 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis. 22160680 2 34 dep donor 459:463 arg1 UDP-glucose 465:475 UDP-glucose 465:475 the sugar donor UDP-glucose 449:475 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 2 35 theme dynamic 374:380 arg1 switch 397:402 a dynamic conformational switch 372:402 a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose 372:475 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 0 36 theme maltosaccharide 48:62 arg1 substrate 64:72 its maltosaccharide substrate 44:72 its maltosaccharide substrate 44:72 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis. 22160680 4 37 theme sugar 852:856 arg1 groups 858:863 sugar groups 852:863 sugar groups 852:863 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 4 38 theme intersubunit 750:761 arg1 mode 763:766 either an intra- or intersubunit mode 730:766 mode 763:766 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 4 39 theme intra- 740:745 arg1 mode 763:766 either an intra- or intersubunit mode 730:766 mode 763:766 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 2 40 theme crystallographic 288:303 arg1 snapshots 305:313 crystallographic snapshots 288:313 crystallographic snapshots of human glycogenin 288:333 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 4 41 theme active 715:720 arg1 site 722:725 the active site 711:725 the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors 711:813 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 1 42 theme maltosaccharide 142:156 arg1 chain 158:162 a maltosaccharide chain 140:162 a maltosaccharide chain covalently attached to itself on Tyr195 140:202 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 6 43 theme catalytic 1148:1156 arg1 mechanism 1158:1166 its catalytic mechanism 1144:1166 its catalytic mechanism 1144:1166 Our data highlight the conformational plasticity of glycogenin and coexistence of two modes of glucosylation as integral to its catalytic mechanism. 22160680 1 44 theme chain 158:162 arg1 synthesis 127:135 the synthesis 123:135 the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 123:202 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 5 45 from locked 965:970 arg1 state 986:990 the ground state 975:990 the ground state 975:990 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 1 46 theme glycogen 251:258 arg1 synthesis 260:268 glycogen synthesis 251:268 glycogen synthesis 251:268 Glycogenin initiates the synthesis of a maltosaccharide chain covalently attached to itself on Tyr195 via a stepwise glucosylation reaction, priming glycogen synthesis. 22160680 0 47 theme glycogen 81:88 arg1 biogenesis 90:99 glycogen biogenesis 81:99 glycogen biogenesis 81:99 Conformational plasticity of glycogenin and its maltosaccharide substrate during glycogen biogenesis. 22160680 5 48 theme glycogen 916:923 arg1 disease 933:939 glycogen storage disease XV 916:942 glycogen storage disease XV 916:942 The Thr83Met mutation, which causes glycogen storage disease XV, is conformationally locked in the ground state and catalytically inactive. 22160680 2 49 theme ground 412:417 arg1 states 430:435 ground and active states 412:435 ground and active states mediated by the sugar donor UDP-glucose 412:475 We have captured crystallographic snapshots of human glycogenin during its reaction cycle, revealing a dynamic conformational switch between ground and active states mediated by the sugar donor UDP-glucose. 22160680 4 50 theme rearranged 650:659 arg1 lid 661:663 The rearranged lid 646:663 The rearranged lid 646:663 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 4 51 dep mode 763:766 arg1 either 730:735 either 730:735 either 730:735 The rearranged lid guides the nascent maltosaccharide chain into the active site in either an intra- or intersubunit mode dependent upon chain length and steric factors and positions the donor and acceptor sugar groups for catalysis. 22160680 3 52 theme major 560:564 arg1 movement 566:573 major movement 560:573 major movement of an approximately 30-residue "lid" segment covering the active site 560:643 This switch includes the ordering of a polypeptide stretch containing Tyr195, and major movement of an approximately 30-residue "lid" segment covering the active site. 34234349 6 0 theme 4,5-bisphosphate 992:1007 arg1 molecule 1009:1016 a phosphatidylinositol 4,5-bisphosphate molecule 969:1016 a phosphatidylinositol 4,5-bisphosphate molecule 969:1016 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 1 1 theme neurotransmitters1-3 207:226 arg1 release 196:202 the release 192:202 the release of neurotransmitters1-3 192:226 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 2 2 theme intractable 325:335 arg1 pain4-6 337:343 intractable pain4-6 325:343 intractable pain4-6 325:343 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 2 3 theme pore 269:272 arg1 Ziconotide 229:238 Ziconotide 229:238 Ziconotide 229:238 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 2 3 theme pore 269:272 arg1 blocker 274:280 a Cav2.2-specific peptide pore blocker 243:280 a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6 243:343 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 4 4 from loops 640:644 arg1 IV 676:677 IV 676:677 IV 676:677 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 4 from loops 640:644 arg1 repeats 656:662 repeats II, III and IV 656:677 repeats II, III and IV of α1 656:683 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 4 from loops 640:644 arg1 II 664:665 II 664:665 II 664:665 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 6 5 theme repeat 869:874 arg1 VSD 862:864 the VSD 858:864 the VSD of repeat II 858:877 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 4 6 from helices 561:567 arg1 IV 676:677 IV 676:677 IV 676:677 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 6 from helices 561:567 arg1 repeats 656:662 repeats II, III and IV 656:677 repeats II, III and IV of α1 656:683 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 6 from helices 561:567 arg1 II 664:665 II 664:665 II 664:665 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 6 7 theme domains 807:813 arg1 VSDs 816:819 VSDs 816:819 VSDs 816:819 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 6 7 theme domains 807:813 arg1 domains 807:813 the voltage-sensing domains 787:813 the voltage-sensing domains (VSDs) 787:820 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 6 7 theme domains 807:813 arg1 Three 778:782 Three 778:782 Three 778:782 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 3 8 theme ziconotide 509:518 arg1 absence 498:504 absence 498:504 absence 498:504 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 3 8 theme ziconotide 509:518 arg1 presence 486:493 presence 486:493 presence 486:493 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 7 9 theme Cav 1182:1184 arg1 channels 1186:1193 Cav channels 1182:1193 Cav channels 1182:1193 Our studies reveal the molecular basis for Cav2.2-specific pore blocking by ziconotide and establish the framework for investigating electromechanical coupling in Cav channels. 34234349 2 10 theme peptide 261:267 arg1 Ziconotide 229:238 Ziconotide 229:238 Ziconotide 229:238 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 2 10 theme peptide 261:267 arg1 blocker 274:280 a Cav2.2-specific peptide pore blocker 243:280 a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6 243:343 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 3 11 theme β3 466:467 arg1 subunits 469:476 the ancillary α2δ-1 and β3 subunits 442:476 subunits 469:476 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 2 12 theme Cav2.2-specific 245:259 arg1 Ziconotide 229:238 Ziconotide 229:238 Ziconotide 229:238 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 2 12 theme Cav2.2-specific 245:259 arg1 blocker 274:280 a Cav2.2-specific peptide pore blocker 243:280 a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6 243:343 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 5 13 theme repeat 724:729 arg1 α2δ-1 739:743 α2δ-1 739:743 α2δ-1 739:743 To accommodate ziconotide, the ECL of repeat III and α2δ-1 have to tilt upward concertedly. 34234349 5 13 theme repeat 724:729 arg1 ECL 717:719 the ECL 713:719 the ECL of repeat III 713:733 To accommodate ziconotide, the ECL of repeat III and α2δ-1 have to tilt upward concertedly. 34234349 6 14 theme phosphatidylinositol 971:990 arg1 4,5-bisphosphate 992:1007 a phosphatidylinositol 4,5-bisphosphate 969:1007 a phosphatidylinositol 4,5-bisphosphate molecule 969:1016 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 3 15 theme core 430:433 arg1 α1 435:436 the core α1 426:436 the core α1 426:436 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 3 16 theme cryo-electron 362:374 arg1 microscopy 376:385 cryo-electron microscopy 362:385 cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) 362:477 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 2 17 used used 307:310 arg2 Ziconotide 229:238 Ziconotide 229:238 Ziconotide 229:238 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 2 17 used used 307:310 arg2 blocker 274:280 a Cav2.2-specific peptide pore blocker 243:280 a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6 243:343 Ziconotide is a Cav2.2-specific peptide pore blocker that has been clinically used for treating intractable pain4-6. 34234349 1 18 theme neuronal-type 76:88 arg1 channels 127:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 6 19 theme voltage-sensing 791:805 arg1 VSDs 816:819 VSDs 816:819 VSDs 816:819 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 6 19 theme voltage-sensing 791:805 arg1 domains 807:813 the voltage-sensing domains 787:813 the voltage-sensing domains (VSDs) 787:820 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 4 20 theme extracellular 626:638 arg1 ECLs 647:650 ECLs 647:650 ECLs 647:650 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 20 theme extracellular 626:638 arg1 loops 640:644 the extracellular loops 622:644 the extracellular loops (ECLs) in repeats II, III and IV of α1 622:683 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 3 21 theme microscopy 376:385 arg1 structures 387:396 cryo-electron microscopy structures 362:396 cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) 362:477 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 7 22 theme Cav2.2-specific 1062:1076 arg1 blocking 1083:1090 Cav2.2-specific pore blocking 1062:1090 Cav2.2-specific pore blocking by ziconotide 1062:1104 Our studies reveal the molecular basis for Cav2.2-specific pore blocking by ziconotide and establish the framework for investigating electromechanical coupling in Cav channels. 34234349 7 23 from coupling 1170:1177 arg1 channels 1186:1193 Cav channels 1182:1193 Cav channels 1182:1193 Our studies reveal the molecular basis for Cav2.2-specific pore blocking by ziconotide and establish the framework for investigating electromechanical coupling in Cav channels. 34234349 6 24 theme intracellular 942:954 arg1 segments 956:963 Cav2-unique intracellular segments 930:963 Cav2-unique intracellular segments 930:963 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 0 25 theme Cav2.2 19:24 arg1 channel 26:32 human Cav2.2 channel 13:32 human Cav2.2 channel 13:32 Structure of human Cav2.2 channel blocked by the painkiller ziconotide. 34234349 1 26 theme N-type 91:96 arg1 channels 127:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 6 27 theme Cav2-unique 930:940 arg1 segments 956:963 Cav2-unique intracellular segments 930:963 Cav2-unique intracellular segments 930:963 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 0 28 theme human 13:17 arg1 channel 26:32 human Cav2.2 channel 13:32 human Cav2.2 channel 13:32 Structure of human Cav2.2 channel blocked by the painkiller ziconotide. 34234349 3 29 dep presence 486:493 arg1 the 482:484 the 482:484 the 482:484 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 3 30 theme ancillary 446:454 arg1 α2δ-1 456:460 the ancillary α2δ-1 and β3 subunits 442:476 α2δ-1 456:460 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 3 31 theme human 401:405 arg1 Cav2.2 407:412 human Cav2.2 401:412 human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) 401:477 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 4 32 dep helices 561:567 arg1 helices 561:567 helices P1 and P2 561:577 helices P1 and P2 561:577 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 32 dep helices 561:567 arg1 P2 576:577 P2 576:577 P2 576:577 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 32 dep helices 561:567 arg1 P1 569:570 P1 569:570 P1 569:570 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 1 33 theme important 174:182 arg1 role 184:187 an important role 171:187 an important role 171:187 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 0 34 theme channel 26:32 arg1 Structure 0:8 Structure 0:8 Structure of human Cav2.2 channel 0:32 Structure of human Cav2.2 channel blocked by the painkiller ziconotide. 34234349 3 35 theme Cav2.2 407:412 arg1 structures 387:396 cryo-electron microscopy structures 362:396 cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) 362:477 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 4 36 theme α1 682:683 arg1 IV 676:677 IV 676:677 IV 676:677 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 36 theme α1 682:683 arg1 repeats 656:662 repeats II, III and IV 656:677 repeats II, III and IV of α1 656:683 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 36 theme α1 682:683 arg1 II 664:665 II 664:665 II 664:665 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 7 37 theme pore 1078:1081 arg1 blocking 1083:1090 Cav2.2-specific pore blocking 1062:1090 Cav2.2-specific pore blocking by ziconotide 1062:1104 Our studies reveal the molecular basis for Cav2.2-specific pore blocking by ziconotide and establish the framework for investigating electromechanical coupling in Cav channels. 34234349 4 38 theme selectivity 598:608 arg1 filter 610:615 the selectivity filter 594:615 the selectivity filter 594:615 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 7 39 theme molecular 1042:1050 arg1 basis 1052:1056 the molecular basis 1038:1056 the molecular basis for Cav2.2-specific pore blocking by ziconotide 1038:1104 Our studies reveal the molecular basis for Cav2.2-specific pore blocking by ziconotide and establish the framework for investigating electromechanical coupling in Cav channels. 34234349 6 40 theme depolarized 831:841 arg1 state 843:847 a depolarized state 829:847 a depolarized state 829:847 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 1 41 theme voltage-gated 99:111 arg1 channels 127:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 1 42 theme calcium 113:119 arg1 channels 127:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 4 43 dep repeats 656:662 arg1 IV 676:677 IV 676:677 IV 676:677 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 43 dep repeats 656:662 arg1 repeats 656:662 repeats II, III and IV 656:677 repeats II, III and IV of α1 656:683 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 4 43 dep repeats 656:662 arg1 II 664:665 II 664:665 II 664:665 Ziconotide is thoroughly coordinated by helices P1 and P2, which support the selectivity filter, and the extracellular loops (ECLs) in repeats II, III and IV of α1. 34234349 3 44 attach present 354:360 arg2 we 351:352 we 351:352 we 351:352 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 3 44 attach present 354:360 arg1 absence 498:504 absence 498:504 absence 498:504 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 3 44 attach present 354:360 arg1 presence 486:493 presence 486:493 presence 486:493 Here we present cryo-electron microscopy structures of human Cav2.2 (comprising the core α1 and the ancillary α2δ-1 and β3 subunits) in the presence or absence of ziconotide. 34234349 1 45 theme Cav 122:124 arg1 channels 127:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 7 46 theme electromechanical 1152:1168 arg1 coupling 1170:1177 electromechanical coupling 1152:1177 electromechanical coupling in Cav channels 1152:1193 Our studies reveal the molecular basis for Cav2.2-specific pore blocking by ziconotide and establish the framework for investigating electromechanical coupling in Cav channels. 34234349 1 47 contain have 166:169 arg2 role 184:187 an important role 171:187 an important role 171:187 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 1 47 contain have 166:169 arg1 channels 127:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels 72:134 The neuronal-type (N-type) voltage-gated calcium (Cav) channels, which are designated Cav2.2, have an important role in the release of neurotransmitters1-3. 34234349 6 48 theme down 890:893 arg1 conformation 895:906 a down conformation 888:906 a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule 888:1016 Three of the voltage-sensing domains (VSDs) are in a depolarized state, whereas the VSD of repeat II exhibits a down conformation that is stabilized by Cav2-unique intracellular segments and a phosphatidylinositol 4,5-bisphosphate molecule. 34234349 0 49 theme painkiller 49:58 arg1 ziconotide 60:69 the painkiller ziconotide 45:69 the painkiller ziconotide 45:69 Structure of human Cav2.2 channel blocked by the painkiller ziconotide. 10978165 3 0 theme Posttranslational 220:236 arg1 processing 238:247 Posttranslational processing 220:247 Posttranslational processing of HCC-1 220:256 Posttranslational processing of HCC-1 was shown to modulate its biological properties. 10978165 4 1 theme different 329:337 arg1 forms 349:353 several different processed forms 321:353 several different processed forms of HCC-1 321:362 In this study several different processed forms of HCC-1 were isolated. 10978165 11 2 theme biological 1403:1412 arg1 activity 1414:1421 its full biological activity 1394:1421 its full biological activity 1394:1421 The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity. 10978165 9 3 theme nonglycosylated 1128:1142 arg1 1-74 1151:1154 1-74 1151:1154 1-74 1151:1154 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 3 theme nonglycosylated 1128:1142 arg1 products 1115:1122 the major products 1105:1122 the major products 1105:1122 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 3 theme nonglycosylated 1128:1142 arg1 HCC-1 1144:1148 nonglycosylated HCC-1 1128:1148 nonglycosylated HCC-1 (1-74) 1128:1155 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 7 4 with O-glycosylation 749:763 arg1 acids 815:819 two N-acetylneuraminic acids 792:819 two N-acetylneuraminic acids 792:819 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 1 5 theme human 158:162 arg1 plasma 164:169 human plasma 158:169 human plasma 158:169 HCC-1 is the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma. 10978165 7 6 from position 768:775 arg1 O-glycosylation 749:763 O-glycosylation 749:763 O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids 749:819 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 7 6 from position 768:775 arg1 galactose 864:872 the disaccharide N-acetylgalactosamine galactose 825:872 the disaccharide N-acetylgalactosamine galactose 825:872 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 10 7 theme glycosylated 1253:1264 arg1 1-74 1273:1276 1-74 1273:1276 1-74 1273:1276 Our data imply that HCC-1 (1-74), HCC-1 (3-74), HCC-1 (4-74) and glycosylated HCC-1 (1-74) circulate in human blood. 10978165 10 7 theme glycosylated 1253:1264 arg1 HCC-1 1266:1270 glycosylated HCC-1 1253:1270 glycosylated HCC-1 (1-74) 1253:1277 Our data imply that HCC-1 (1-74), HCC-1 (3-74), HCC-1 (4-74) and glycosylated HCC-1 (1-74) circulate in human blood. 10978165 5 8 theme HCC-1 437:441 arg1 band 465:468 a HCC-1 immunoreactive double band 435:468 a HCC-1 immunoreactive double band at 8-10 kDa 435:480 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 5 9 theme Western 379:385 arg1 analysis 392:399 Western blot analysis 379:399 Western blot analysis of human plasma extracts 379:424 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 5 10 theme HCC-1 522:526 arg1 peptides 528:535 two distinct HCC-1 peptides 509:535 two distinct HCC-1 peptides 509:535 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 7 11 theme N-acetylgalactosamine 842:862 arg1 galactose 864:872 the disaccharide N-acetylgalactosamine galactose 825:872 the disaccharide N-acetylgalactosamine galactose 825:872 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 5 12 theme blot 387:390 arg1 analysis 392:399 Western blot analysis 379:399 Western blot analysis of human plasma extracts 379:424 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 9 13 gly nonglycosylated 1128:1142 arg1 1-74 1151:1154 1-74 1151:1154 1-74 1151:1154 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 13 gly nonglycosylated 1128:1142 arg1 products 1115:1122 the major products 1105:1122 the major products 1105:1122 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 13 gly nonglycosylated 1128:1142 arg1 HCC-1 1144:1148 nonglycosylated HCC-1 1128:1148 nonglycosylated HCC-1 (1-74) 1128:1155 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 6 14 gly glycosylated 659:670 arg1 1-74 679:682 1-74 679:682 1-74 679:682 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 6 14 gly glycosylated 659:670 arg1 HCC-1 672:676 glycosylated HCC-1 659:676 glycosylated HCC-1 (1-74) 659:683 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 1 15 from concentrations 140:153 arg1 plasma 164:169 human plasma 158:169 human plasma 158:169 HCC-1 is the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma. 10978165 7 16 theme disaccharide 829:840 arg1 galactose 864:872 the disaccharide N-acetylgalactosamine galactose 825:872 the disaccharide N-acetylgalactosamine galactose 825:872 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 7 17 theme molecular 716:724 arg1 mass 726:729 a molecular mass 714:729 a molecular mass of 9621 Da 714:740 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 9 18 gly glycosylated 1161:1172 arg1 1-74 1181:1184 1-74 1181:1184 1-74 1181:1184 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 18 gly glycosylated 1161:1172 arg1 HCC-1 1174:1178 glycosylated HCC-1 1161:1178 glycosylated HCC-1 (1-74) 1161:1185 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 6 19 attach isolated 558:565 arg1 library 582:588 a peptide library 572:588 a peptide library of human blood filtrate 572:612 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 6 19 attach isolated 558:565 arg2 peptides 544:551 These peptides 538:551 These peptides 538:551 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 8 20 theme truncated 900:908 arg1 3-74 917:920 3-74 917:920 3-74 917:920 Furthermore N-terminally truncated HCC-1 (3-74) and HCC-1 (4-74) were identified in the peptide library. 10978165 8 20 theme truncated 900:908 arg1 HCC-1 910:914 N-terminally truncated HCC-1 887:914 N-terminally truncated HCC-1 (3-74) 887:921 Furthermore N-terminally truncated HCC-1 (3-74) and HCC-1 (4-74) were identified in the peptide library. 10978165 0 21 theme processed 20:28 arg1 forms 30:34 Posttranslationally processed forms 0:34 Posttranslationally processed forms of the human chemokine HCC-1 0:63 Posttranslationally processed forms of the human chemokine HCC-1. 10978165 9 22 theme glycosylated 1161:1172 arg1 1-74 1181:1184 1-74 1181:1184 1-74 1181:1184 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 22 theme glycosylated 1161:1172 arg1 HCC-1 1174:1178 glycosylated HCC-1 1161:1178 glycosylated HCC-1 (1-74) 1161:1185 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 1 23 theme only 79:82 arg1 CC-chemokine 84:95 the only CC-chemokine 75:95 the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma 75:169 HCC-1 is the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma. 10978165 1 23 theme only 79:82 arg1 HCC-1 66:70 HCC-1 66:70 HCC-1 66:70 HCC-1 is the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma. 10978165 5 24 theme distinct 513:520 arg1 peptides 528:535 two distinct HCC-1 peptides 509:535 two distinct HCC-1 peptides 509:535 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 6 25 theme filtrate 605:612 arg1 library 582:588 a peptide library 572:588 a peptide library of human blood filtrate 572:612 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 3 26 theme HCC-1 252:256 arg1 processing 238:247 Posttranslational processing 220:247 Posttranslational processing of HCC-1 220:256 Posttranslational processing of HCC-1 was shown to modulate its biological properties. 10978165 10 27 theme human 1292:1296 arg1 blood 1298:1302 human blood 1292:1302 human blood 1292:1302 Our data imply that HCC-1 (1-74), HCC-1 (3-74), HCC-1 (4-74) and glycosylated HCC-1 (1-74) circulate in human blood. 10978165 11 28 theme HCC-1 1351:1355 arg1 processing 1320:1329 N-terminal processing 1309:1329 N-terminal processing 1309:1329 The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity. 10978165 11 28 theme HCC-1 1351:1355 arg1 modification 1335:1346 modification 1335:1346 modification 1335:1346 The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity. 10978165 6 29 theme blood 599:603 arg1 filtrate 605:612 human blood filtrate 593:612 human blood filtrate 593:612 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 30 theme major 1109:1113 arg1 products 1115:1122 the major products 1105:1122 the major products 1105:1122 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 30 theme major 1109:1113 arg1 HCC-1 1144:1148 nonglycosylated HCC-1 1128:1148 nonglycosylated HCC-1 (1-74) 1128:1155 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 8 31 theme peptide 963:969 arg1 library 971:977 the peptide library 959:977 the peptide library 959:977 Furthermore N-terminally truncated HCC-1 (3-74) and HCC-1 (4-74) were identified in the peptide library. 10978165 6 32 theme human 593:597 arg1 filtrate 605:612 human blood filtrate 593:612 human blood filtrate 593:612 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 3 33 theme biological 284:293 arg1 properties 295:304 its biological properties 280:304 its biological properties 280:304 Posttranslational processing of HCC-1 was shown to modulate its biological properties. 10978165 2 34 theme physiological 176:188 arg1 function 190:197 Its physiological function 172:197 Its physiological function 172:197 Its physiological function is not well defined. 10978165 0 35 theme human 43:47 arg1 HCC-1 59:63 the human chemokine HCC-1 39:63 the human chemokine HCC-1 39:63 Posttranslationally processed forms of the human chemokine HCC-1. 10978165 7 36 theme N-acetylneuraminic 796:813 arg1 acids 815:819 two N-acetylneuraminic acids 792:819 two N-acetylneuraminic acids 792:819 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 6 37 theme glycosylated 659:670 arg1 1-74 679:682 1-74 679:682 1-74 679:682 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 6 37 theme glycosylated 659:670 arg1 HCC-1 672:676 glycosylated HCC-1 659:676 glycosylated HCC-1 (1-74) 659:683 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 11 38 theme full 1398:1401 arg1 activity 1414:1421 its full biological activity 1394:1421 its full biological activity 1394:1421 The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity. 10978165 0 39 theme HCC-1 59:63 arg1 forms 30:34 Posttranslationally processed forms 0:34 Posttranslationally processed forms of the human chemokine HCC-1 0:63 Posttranslationally processed forms of the human chemokine HCC-1. 10978165 9 40 dep % 1071:1071 arg1 represents 1073:1082 represents 1073:1082 represents HCC-1 (4-74) 1073:1095 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 7 41 theme Glycosylated 686:697 arg1 HCC-1 699:703 Glycosylated HCC-1 686:703 Glycosylated HCC-1 686:703 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 0 42 theme chemokine 49:57 arg1 HCC-1 59:63 the human chemokine HCC-1 39:63 the human chemokine HCC-1 39:63 Posttranslationally processed forms of the human chemokine HCC-1. 10978165 11 43 theme N-terminal 1309:1318 arg1 processing 1320:1329 N-terminal processing 1309:1329 N-terminal processing 1309:1329 The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity. 10978165 5 44 from kDa 478:480 arg1 band 465:468 a HCC-1 immunoreactive double band 435:468 a HCC-1 immunoreactive double band at 8-10 kDa 435:480 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 11 45 dep processing 1320:1329 arg1 The 1305:1307 The 1305:1307 The 1305:1307 The N-terminal processing and modification of HCC-1 might be of importance in displaying its full biological activity. 10978165 5 46 theme immunoreactive 443:456 arg1 band 465:468 a HCC-1 immunoreactive double band 435:468 a HCC-1 immunoreactive double band at 8-10 kDa 435:480 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 4 47 theme HCC-1 358:362 arg1 forms 349:353 several different processed forms 321:353 several different processed forms of HCC-1 321:362 In this study several different processed forms of HCC-1 were isolated. 10978165 5 48 theme double 458:463 arg1 band 465:468 a HCC-1 immunoreactive double band 435:468 a HCC-1 immunoreactive double band at 8-10 kDa 435:480 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 7 49 gly O-glycosylation 749:763 arg1 position 768:775 position 7 768:777 position 7 (Ser-7) 768:785 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 7 49 gly O-glycosylation 749:763 arg1 Ser-7 780:784 Ser-7 780:784 Ser-7 780:784 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 7 49 gly O-glycosylation 749:763 arg2 Ser-7 780:784 Ser-7 780:784 Ser-7 780:784 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 7 49 gly O-glycosylation 749:763 arg2 position 768:775 position 7 768:777 position 7 (Ser-7) 768:785 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 4 50 theme several 321:327 arg1 forms 349:353 several different processed forms 321:353 several different processed forms of HCC-1 321:362 In this study several different processed forms of HCC-1 were isolated. 10978165 9 51 theme total 1016:1020 arg1 HCC-1 1022:1026 total HCC-1 1016:1026 total HCC-1 1016:1026 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 10 52 gly glycosylated 1253:1264 arg1 1-74 1273:1276 1-74 1273:1276 1-74 1273:1276 Our data imply that HCC-1 (1-74), HCC-1 (3-74), HCC-1 (4-74) and glycosylated HCC-1 (1-74) circulate in human blood. 10978165 10 52 gly glycosylated 1253:1264 arg1 HCC-1 1266:1270 glycosylated HCC-1 1253:1270 glycosylated HCC-1 (1-74) 1253:1277 Our data imply that HCC-1 (1-74), HCC-1 (3-74), HCC-1 (4-74) and glycosylated HCC-1 (1-74) circulate in human blood. 10978165 7 53 with galactose 864:872 arg1 acids 815:819 two N-acetylneuraminic acids 792:819 two N-acetylneuraminic acids 792:819 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 5 54 theme peptides 528:535 arg1 presence 497:504 the presence 493:504 the presence of two distinct HCC-1 peptides 493:535 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 1 55 theme nanomolar 130:138 arg1 concentrations 140:153 nanomolar concentrations 130:153 nanomolar concentrations in human plasma 130:169 HCC-1 is the only CC-chemokine known so far which circulates in nanomolar concentrations in human plasma. 10978165 9 56 theme HCC-1 1022:1026 arg1 % 1011:1011 approximately 3% 996:1011 approximately 3% of total HCC-1 996:1026 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 9 56 theme HCC-1 1022:1026 arg1 HCC-1 1022:1026 total HCC-1 1016:1026 total HCC-1 1016:1026 In hemofiltrate approximately 3% of total HCC-1 represents HCC-1 (3-74) and approximately 1% represents HCC-1 (4-74) whereas the major products are nonglycosylated HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 5 57 theme human 404:408 arg1 extracts 417:424 human plasma extracts 404:424 human plasma extracts 404:424 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 6 58 theme peptide 574:580 arg1 library 582:588 a peptide library 572:588 a peptide library of human blood filtrate 572:612 These peptides were isolated from a peptide library of human blood filtrate and represent predominantly HCC-1 (1-74) and glycosylated HCC-1 (1-74). 10978165 7 59 theme Da 739:740 arg1 mass 726:729 a molecular mass 714:729 a molecular mass of 9621 Da 714:740 Glycosylated HCC-1 exhibits a molecular mass of 9621 Da due to O-glycosylation at position 7 (Ser-7) with two N-acetylneuraminic acids and the disaccharide N-acetylgalactosamine galactose. 10978165 4 60 theme processed 339:347 arg1 forms 349:353 several different processed forms 321:353 several different processed forms of HCC-1 321:362 In this study several different processed forms of HCC-1 were isolated. 10978165 5 61 theme extracts 417:424 arg1 analysis 392:399 Western blot analysis 379:399 Western blot analysis of human plasma extracts 379:424 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 10978165 5 62 theme plasma 410:415 arg1 extracts 417:424 human plasma extracts 404:424 human plasma extracts 404:424 Western blot analysis of human plasma extracts revealed a HCC-1 immunoreactive double band at 8-10 kDa indicating the presence of two distinct HCC-1 peptides. 23056909 3 0 theme physiological 685:697 arg1 bradykinin 632:641 vasodilator bradykinin 620:641 vasodilator bradykinin 620:641 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 0 theme physiological 685:697 arg1 modulator 699:707 a physiological modulator 683:707 a physiological modulator of hematopoiesis 683:724 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 0 1 theme ACE 79:81 arg1 activity 84:91 human angiotensin-I converting enzyme (ACE) activity 40:91 human angiotensin-I converting enzyme (ACE) activity 40:91 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 5 2 theme renin-angiotensin 1079:1095 arg1 RAS 1105:1107 RAS 1105:1107 RAS 1105:1107 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 2 theme renin-angiotensin 1079:1095 arg1 system 1097:1102 the human renin-angiotensin system 1069:1102 the human renin-angiotensin system (RAS) 1069:1108 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 3 theme human 1007:1011 arg1 ACE 1021:1023 human somatic ACE 1007:1023 human somatic ACE 1007:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 4 4 theme natural 835:841 arg1 Ang 908:910 Ang II 908:913 Ang II 908:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 4 theme natural 835:841 arg1 b 902:902 bradykinin potentiating peptide b 870:902 bradykinin potentiating peptide b 870:902 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 4 theme natural 835:841 arg1 inhibitors 851:860 the natural peptide inhibitors 831:860 the natural peptide inhibitors of ACE 831:867 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 3 5 theme octapeptide 513:523 arg1 angiotensin 525:535 the vasoactive octapeptide angiotensin II 498:538 the vasoactive octapeptide angiotensin II (Ang II) 498:547 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 5 theme octapeptide 513:523 arg1 Ang 541:543 Ang II 541:546 Ang II 541:546 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 0 6 theme activity 84:91 arg1 regulation 26:35 regulation 26:35 regulation 26:35 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 0 6 theme activity 84:91 arg1 recognition 10:20 Molecular recognition 0:20 Molecular recognition 0:20 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 1 7 from systems 333:339 arg1 result 247:252 a result 245:252 a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems 245:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 5 8 theme regulatory 1050:1059 arg1 role 1061:1064 a regulatory role 1048:1064 a regulatory role in the human renin-angiotensin system (RAS) 1048:1108 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 1 9 from role 270:273 arg1 systems 333:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 4 10 theme first 735:739 arg1 time 741:744 the first time 731:744 the first time 731:744 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 0 11 theme natural 96:102 arg1 peptides 115:122 natural inhibitory peptides 96:122 natural inhibitory peptides 96:122 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 5 12 theme human 1073:1077 arg1 RAS 1105:1107 RAS 1105:1107 RAS 1105:1107 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 12 theme human 1073:1077 arg1 system 1097:1102 the human renin-angiotensin system 1069:1102 the human renin-angiotensin system (RAS) 1069:1108 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 3 13 theme vasoactive 502:511 arg1 angiotensin 525:535 the vasoactive octapeptide angiotensin II 498:538 the vasoactive octapeptide angiotensin II (Ang II) 498:547 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 13 theme vasoactive 502:511 arg1 Ang 541:543 Ang II 541:546 Ang II 541:546 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 0 14 theme inhibitory 104:113 arg1 peptides 115:122 natural inhibitory peptides 96:122 natural inhibitory peptides 96:122 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 5 15 from role 1061:1064 arg1 RAS 1105:1107 RAS 1105:1107 RAS 1105:1107 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 15 from role 1061:1064 arg1 system 1097:1102 the human renin-angiotensin system 1069:1102 the human renin-angiotensin system (RAS) 1069:1108 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 4 16 theme domain 809:814 arg1 selectivity 816:826 the domain selectivity 805:826 the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II 805:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 1 17 theme two-domain 166:175 arg1 dipeptidylcarboxypeptidase 177:202 a two-domain dipeptidylcarboxypeptidase 164:202 a two-domain dipeptidylcarboxypeptidase 164:202 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 17 theme two-domain 166:175 arg1 enzyme 150:155 Angiotensin-I converting enzyme 125:155 Angiotensin-I converting enzyme (ACE) 125:161 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 0 18 theme Molecular 0:8 arg1 recognition 10:20 Molecular recognition 0:20 Molecular recognition 0:20 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 3 19 dep bradykinin 632:641 arg1 the 616:618 the 616:618 the 616:618 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 5 20 theme somatic 1013:1019 arg1 ACE 1021:1023 human somatic ACE 1007:1023 human somatic ACE 1007:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 1 21 theme critical 261:268 arg1 role 270:273 its critical role 257:273 its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems 257:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 22 from result 247:252 arg1 systems 333:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 23 theme role 270:273 arg1 result 247:252 a result 245:252 a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems 245:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 5 24 theme ACE 1021:1023 arg1 domain 997:1002 the carboxy-terminal domain 976:1002 the carboxy-terminal domain of human somatic ACE 976:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 24 theme ACE 1021:1023 arg1 ACE 1021:1023 human somatic ACE 1007:1023 human somatic ACE 1007:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 2 25 theme diseases 414:421 arg1 treatment 386:394 the treatment 382:394 the treatment of cardiovascular diseases 382:421 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 0 26 theme human 40:44 arg1 activity 84:91 human angiotensin-I converting enzyme (ACE) activity 40:91 human angiotensin-I converting enzyme (ACE) activity 40:91 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 4 27 theme peptide 894:900 arg1 b 902:902 bradykinin potentiating peptide b 870:902 bradykinin potentiating peptide b 870:902 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 27 theme peptide 894:900 arg1 inhibitors 851:860 the natural peptide inhibitors 831:860 the natural peptide inhibitors of ACE 831:867 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 3 28 theme other 589:593 arg1 N-acetyl-Ser-Asp-Lys-Pro 647:670 N-acetyl-Ser-Asp-Lys-Pro 647:670 N-acetyl-Ser-Asp-Lys-Pro 647:670 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 28 theme other 589:593 arg1 bradykinin 632:641 vasodilator bradykinin 620:641 vasodilator bradykinin 620:641 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 28 theme other 589:593 arg1 substrates 595:604 other substrates 589:604 other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis 589:724 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 4 29 theme biochemical 768:778 arg1 basis 795:799 a detailed biochemical and structural basis 757:799 a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II 757:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 2 30 theme cardiovascular 399:412 arg1 diseases 414:421 cardiovascular diseases 399:421 cardiovascular diseases 399:421 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 4 31 theme potentiating 881:892 arg1 b 902:902 bradykinin potentiating peptide b 870:902 bradykinin potentiating peptide b 870:902 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 31 theme potentiating 881:892 arg1 inhibitors 851:860 the natural peptide inhibitors 831:860 the natural peptide inhibitors of ACE 831:867 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 32 theme detailed 759:766 arg1 basis 795:799 a detailed biochemical and structural basis 757:799 a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II 757:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 1 33 theme key 210:212 arg1 regulator 214:222 a key regulator 208:222 a key regulator of blood pressure 208:240 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 33 theme key 210:212 arg1 enzyme 150:155 Angiotensin-I converting enzyme 125:155 Angiotensin-I converting enzyme (ACE) 125:161 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 0 34 theme converting 60:69 arg1 activity 84:91 human angiotensin-I converting enzyme (ACE) activity 40:91 human angiotensin-I converting enzyme (ACE) activity 40:91 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 1 35 theme renin-angiotensin-aldosterone 282:310 arg1 systems 333:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 4 36 theme inhibitors 851:860 arg1 selectivity 816:826 the domain selectivity 805:826 the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II 805:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 3 37 theme hematopoiesis 712:724 arg1 bradykinin 632:641 vasodilator bradykinin 620:641 vasodilator bradykinin 620:641 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 37 theme hematopoiesis 712:724 arg1 modulator 699:707 a physiological modulator 683:707 a physiological modulator of hematopoiesis 683:724 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 4 38 theme structural 784:793 arg1 basis 795:799 a detailed biochemical and structural basis 757:799 a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II 757:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 2 39 from target 372:377 arg1 treatment 386:394 the treatment 382:394 the treatment of cardiovascular diseases 382:421 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 0 40 theme angiotensin-I 46:58 arg1 activity 84:91 human angiotensin-I converting enzyme (ACE) activity 40:91 human angiotensin-I converting enzyme (ACE) activity 40:91 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 4 41 theme peptide 843:849 arg1 Ang 908:910 Ang II 908:913 Ang II 908:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 41 theme peptide 843:849 arg1 b 902:902 bradykinin potentiating peptide b 870:902 bradykinin potentiating peptide b 870:902 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 41 theme peptide 843:849 arg1 inhibitors 851:860 the natural peptide inhibitors 831:860 the natural peptide inhibitors of ACE 831:867 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 5 42 theme domain 997:1002 arg1 inhibition 962:971 selective competitive inhibition 940:971 selective competitive inhibition of the carboxy-terminal domain of human somatic ACE 940:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 43 theme carboxy-terminal 980:995 arg1 domain 997:1002 the carboxy-terminal domain 976:1002 the carboxy-terminal domain of human somatic ACE 976:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 5 43 theme carboxy-terminal 980:995 arg1 ACE 1021:1023 human somatic ACE 1007:1023 human somatic ACE 1007:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 1 44 theme kallikrein-kinin 316:331 arg1 systems 333:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 the renin-angiotensin-aldosterone and kallikrein-kinin systems 278:339 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 4 45 theme ACE 865:867 arg1 Ang 908:910 Ang II 908:913 Ang II 908:913 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 45 theme ACE 865:867 arg1 b 902:902 bradykinin potentiating peptide b 870:902 bradykinin potentiating peptide b 870:902 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 45 theme ACE 865:867 arg1 inhibitors 851:860 the natural peptide inhibitors 831:860 the natural peptide inhibitors of ACE 831:867 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 3 46 theme vasodilator 620:630 arg1 bradykinin 632:641 vasodilator bradykinin 620:641 vasodilator bradykinin 620:641 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 46 theme vasodilator 620:630 arg1 modulator 699:707 a physiological modulator 683:707 a physiological modulator of hematopoiesis 683:724 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 46 theme vasodilator 620:630 arg1 Ac-SDKP 673:679 Ac-SDKP 673:679 Ac-SDKP 673:679 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 1 47 theme Angiotensin-I 125:137 arg1 regulator 214:222 a key regulator 208:222 a key regulator of blood pressure 208:240 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 47 theme Angiotensin-I 125:137 arg1 dipeptidylcarboxypeptidase 177:202 a two-domain dipeptidylcarboxypeptidase 164:202 a two-domain dipeptidylcarboxypeptidase 164:202 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 47 theme Angiotensin-I 125:137 arg1 ACE 158:160 ACE 158:160 ACE 158:160 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 47 theme Angiotensin-I 125:137 arg1 enzyme 150:155 Angiotensin-I converting enzyme 125:155 Angiotensin-I converting enzyme (ACE) 125:161 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 5 48 dep showed 933:938 arg1 providing 1025:1033 providing 1025:1033 showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS) 933:1108 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 3 49 theme substrates 595:604 arg1 N-acetyl-Ser-Asp-Lys-Pro 647:670 N-acetyl-Ser-Asp-Lys-Pro 647:670 N-acetyl-Ser-Asp-Lys-Pro 647:670 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 49 theme substrates 595:604 arg1 bradykinin 632:641 vasodilator bradykinin 620:641 vasodilator bradykinin 620:641 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 3 49 theme substrates 595:604 arg1 number 579:584 a number 577:584 a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis 577:724 ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. 23056909 1 50 theme converting 139:148 arg1 regulator 214:222 a key regulator 208:222 a key regulator of blood pressure 208:240 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 50 theme converting 139:148 arg1 dipeptidylcarboxypeptidase 177:202 a two-domain dipeptidylcarboxypeptidase 164:202 a two-domain dipeptidylcarboxypeptidase 164:202 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 50 theme converting 139:148 arg1 ACE 158:160 ACE 158:160 ACE 158:160 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 50 theme converting 139:148 arg1 enzyme 150:155 Angiotensin-I converting enzyme 125:155 Angiotensin-I converting enzyme (ACE) 125:161 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 4 51 theme bradykinin 870:879 arg1 b 902:902 bradykinin potentiating peptide b 870:902 bradykinin potentiating peptide b 870:902 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 4 51 theme bradykinin 870:879 arg1 inhibitors 851:860 the natural peptide inhibitors 831:860 the natural peptide inhibitors of ACE 831:867 For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. 23056909 5 52 theme selective 940:948 arg1 inhibition 962:971 selective competitive inhibition 940:971 selective competitive inhibition of the carboxy-terminal domain of human somatic ACE 940:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 2 53 theme drug 367:370 arg1 target 372:377 an important drug target 354:377 an important drug target in the treatment of cardiovascular diseases 354:421 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 2 53 theme drug 367:370 arg1 it 348:349 it 348:349 it 348:349 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 1 54 theme blood 227:231 arg1 pressure 233:240 blood pressure 227:240 blood pressure 227:240 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 0 55 theme enzyme 71:76 arg1 activity 84:91 human angiotensin-I converting enzyme (ACE) activity 40:91 human angiotensin-I converting enzyme (ACE) activity 40:91 Molecular recognition and regulation of human angiotensin-I converting enzyme (ACE) activity by natural inhibitory peptides. 23056909 5 56 theme competitive 950:960 arg1 inhibition 962:971 selective competitive inhibition 940:971 selective competitive inhibition of the carboxy-terminal domain of human somatic ACE 940:1023 Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS). 23056909 2 57 theme important 357:365 arg1 target 372:377 an important drug target 354:377 an important drug target in the treatment of cardiovascular diseases 354:421 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 2 57 theme important 357:365 arg1 it 348:349 it 348:349 it 348:349 Hence it is an important drug target in the treatment of cardiovascular diseases. 23056909 1 58 theme pressure 233:240 arg1 regulator 214:222 a key regulator 208:222 a key regulator of blood pressure 208:240 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 23056909 1 58 theme pressure 233:240 arg1 enzyme 150:155 Angiotensin-I converting enzyme 125:155 Angiotensin-I converting enzyme (ACE) 125:161 Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. 27551080 0 0 theme NPC1 69:72 arg1 domain 89:94 NPC1 middle lumenal domain 69:94 NPC1 middle lumenal domain 69:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 5 1 theme protruding 661:670 arg1 loops 672:676 two protruding loops 657:676 two protruding loops 657:676 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 5 2 gly glycoprotein 749:760 arg1 glycoprotein 749:760 the primed Ebola virus glycoprotein 726:760 the primed Ebola virus glycoprotein 726:760 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 1 3 theme cholesterol 133:143 arg1 Export 111:116 Export 111:116 Export of LDL-derived cholesterol from lysosomes 111:158 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 6 4 theme cholesterol 898:908 arg1 pockets 918:924 cholesterol binding pockets 898:924 cholesterol binding pockets 898:924 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 6 5 theme full-length 805:815 arg1 structure 822:830 the full-length NPC1 structure 801:830 the full-length NPC1 structure 801:830 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 2 6 from Mutations 288:296 arg1 genes 305:309 the genes 301:309 the genes encoding these proteins 301:333 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 0 7 theme lumenal 81:87 arg1 domain 89:94 NPC1 middle lumenal domain 69:94 NPC1 middle lumenal domain 69:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 0 8 theme middle 74:79 arg1 domain 89:94 NPC1 middle lumenal domain 69:94 NPC1 middle lumenal domain 69:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 6 9 theme complex 788:794 arg1 Docking 763:769 Docking 763:769 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure 763:830 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 4 10 theme human 580:584 arg1 NPC1-MLD 586:593 NPC1-MLD 586:593 NPC1-MLD 586:593 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 5 11 theme virus 743:747 arg1 glycoprotein 749:760 the primed Ebola virus glycoprotein 726:760 the primed Ebola virus glycoprotein 726:760 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 6 12 theme NPC1-NPC2 778:786 arg1 complex 788:794 the NPC1-NPC2 complex 774:794 the NPC1-NPC2 complex 774:794 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 6 13 theme hydrophobic 943:953 arg1 model 986:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 2 14 theme disease 356:362 arg1 C 369:369 Niemann-Pick disease type C 343:369 Niemann-Pick disease type C (NPC) 343:375 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 2 14 theme disease 356:362 arg1 NPC 372:374 NPC 372:374 NPC 372:374 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 0 15 theme domain 89:94 arg1 structure 56:64 the structure 52:64 the structure of NPC1 middle lumenal domain 52:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 6 16 dep NPC2 885:888 arg1 pockets 918:924 cholesterol binding pockets 898:924 cholesterol binding pockets 898:924 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 2 17 theme Niemann-Pick 343:354 arg1 C 369:369 Niemann-Pick disease type C 343:369 Niemann-Pick disease type C (NPC) 343:375 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 2 17 theme Niemann-Pick 343:354 arg1 NPC 372:374 NPC 372:374 NPC 372:374 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 5 18 theme analogous 692:700 arg1 NPC2 686:689 NPC2 686:689 NPC2 686:689 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 6 19 theme cholesterol 849:859 arg1 tunnel 870:875 a direct cholesterol transfer tunnel 840:875 a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets 840:924 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 6 20 theme cholesterol 965:975 arg1 model 986:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 1 21 theme soluble 247:253 arg1 protein 255:261 a soluble protein 245:261 a soluble protein 245:261 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 21 theme soluble 247:253 arg1 C2 277:278 Niemann-Pick C2 264:278 Niemann-Pick C2 (NPC2) 264:285 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 4 22 theme bound 612:616 arg1 cholesterol-3-O-sulfate 618:640 bound cholesterol-3-O-sulfate 612:640 bound cholesterol-3-O-sulfate 612:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 6 23 theme " 963:963 arg1 model 986:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 5 24 theme primed 730:735 arg1 glycoprotein 749:760 the primed Ebola virus glycoprotein 726:760 the primed Ebola virus glycoprotein 726:760 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 1 25 theme protein 255:261 arg1 cooperation 173:183 the cooperation 169:183 the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2) 169:285 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 6 26 theme transfer 977:984 arg1 model 986:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 6 27 theme hand-off 955:962 arg1 model 986:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 the "hydrophobic hand-off" cholesterol transfer model 938:990 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 5 28 with interaction 709:719 arg1 glycoprotein 749:760 the primed Ebola virus glycoprotein 726:760 the primed Ebola virus glycoprotein 726:760 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 6 29 theme transfer 861:868 arg1 tunnel 870:875 a direct cholesterol transfer tunnel 840:875 a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets 840:924 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 3 30 theme second 399:404 arg1 MLD 443:445 MLD 443:445 MLD 443:445 NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). 27551080 3 30 theme second 399:404 arg1 domain 435:440 NPC1's second (middle), lumenally oriented domain 392:440 NPC1's second (middle), lumenally oriented domain (MLD) 392:446 NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). 27551080 0 31 theme cholesterol 26:36 arg1 transfer 38:45 cholesterol transfer 26:45 cholesterol transfer from the structure of NPC1 middle lumenal domain 26:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 1 32 theme Niemann-Pick 264:275 arg1 NPC2 281:284 NPC2 281:284 NPC2 281:284 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 32 theme Niemann-Pick 264:275 arg1 protein 255:261 a soluble protein 245:261 a soluble protein 245:261 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 32 theme Niemann-Pick 264:275 arg1 C2 277:278 Niemann-Pick C2 264:278 Niemann-Pick C2 (NPC2) 264:285 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 4 33 theme 2.4-Å 529:533 arg1 structure 554:562 the 2.4-Å resolution crystal structure 525:562 the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate 525:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 1 34 theme integral 192:199 arg1 protein 210:216 the integral membrane protein 188:216 the integral membrane protein Niemann-Pick C1 (NPC1) 188:239 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 34 theme integral 192:199 arg1 NPC1 235:238 NPC1 235:238 NPC1 235:238 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 4 35 theme crystal 546:552 arg1 structure 554:562 the 2.4-Å resolution crystal structure 525:562 the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate 525:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 6 36 theme direct 842:847 arg1 tunnel 870:875 a direct cholesterol transfer tunnel 840:875 a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets 840:924 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 1 37 link LDL-derived 121:131 arg1 cholesterol 133:143 LDL-derived cholesterol 121:143 LDL-derived cholesterol 121:143 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 38 dep protein 210:216 arg1 C1 231:232 Niemann-Pick C1 218:232 the integral membrane protein Niemann-Pick C1 (NPC1) 188:239 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 6 39 theme binding 910:916 arg1 pockets 918:924 cholesterol binding pockets 898:924 cholesterol binding pockets 898:924 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 5 40 theme Ebola 737:741 arg1 glycoprotein 749:760 the primed Ebola virus glycoprotein 726:760 the primed Ebola virus glycoprotein 726:760 NPC1-MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. 27551080 1 41 theme membrane 201:208 arg1 protein 210:216 the integral membrane protein 188:216 the integral membrane protein Niemann-Pick C1 (NPC1) 188:239 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 41 theme membrane 201:208 arg1 NPC1 235:238 NPC1 235:238 NPC1 235:238 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 3 42 theme N-terminal 484:493 arg1 NTD 503:505 NTD 503:505 NTD 503:505 NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). 27551080 3 42 theme N-terminal 484:493 arg1 domain 495:500 NPC1's N-terminal domain 477:500 NPC1's N-terminal domain (NTD) 477:506 NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). 27551080 0 43 theme transfer 38:45 arg1 mechanism 13:21 the mechanism 9:21 the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain 9:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 6 44 theme NPC1 817:820 arg1 structure 822:830 the full-length NPC1 structure 801:830 the full-length NPC1 structure 801:830 Docking of the NPC1-NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the "hydrophobic hand-off" cholesterol transfer model. 27551080 0 45 from structure 56:64 arg1 mechanism 13:21 the mechanism 9:21 the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain 9:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 0 45 from structure 56:64 arg1 transfer 38:45 cholesterol transfer 26:45 cholesterol transfer from the structure of NPC1 middle lumenal domain 26:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 1 46 theme protein 210:216 arg1 cooperation 173:183 the cooperation 169:183 the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2) 169:285 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 4 47 theme NPC2 599:602 arg1 complex 569:575 a complex 567:575 a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate 567:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 1 48 theme Niemann-Pick 218:229 arg1 C1 231:232 Niemann-Pick C1 218:232 the integral membrane protein Niemann-Pick C1 (NPC1) 188:239 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 2 49 theme type 364:367 arg1 C 369:369 Niemann-Pick disease type C 343:369 Niemann-Pick disease type C (NPC) 343:375 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 2 49 theme type 364:367 arg1 NPC 372:374 NPC 372:374 NPC 372:374 Mutations in the genes encoding these proteins lead to Niemann-Pick disease type C (NPC). 27551080 4 50 theme complex 569:575 arg1 structure 554:562 the 2.4-Å resolution crystal structure 525:562 the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate 525:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 4 51 theme resolution 535:544 arg1 structure 554:562 the 2.4-Å resolution crystal structure 525:562 the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate 525:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 27551080 0 52 from mechanism 13:21 arg1 structure 56:64 the structure 52:64 the structure of NPC1 middle lumenal domain 52:94 Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. 27551080 3 53 dep second 399:404 arg1 oriented 426:433 oriented 426:433 oriented 426:433 NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). 27551080 3 53 dep second 399:404 arg1 middle 407:412 middle 407:412 middle 407:412 NPC2 binds to NPC1's second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1's N-terminal domain (NTD). 27551080 1 54 from lysosomes 150:158 arg1 Export 111:116 Export 111:116 Export of LDL-derived cholesterol from lysosomes 111:158 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 1 55 theme LDL-derived 121:131 arg1 cholesterol 133:143 LDL-derived cholesterol 121:143 LDL-derived cholesterol 121:143 Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann-Pick C1 (NPC1) and a soluble protein, Niemann-Pick C2 (NPC2). 27551080 4 56 theme NPC1-MLD 586:593 arg1 complex 569:575 a complex 567:575 a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate 567:640 Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1-MLD and NPC2 bearing bound cholesterol-3-O-sulfate. 28165004 6 0 theme functional 694:703 arg1 analyses 705:712 Structural and functional analyses 679:712 Structural and functional analyses 679:712 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 4 1 theme PD-1 477:480 arg1 glycosylation 482:494 PD-1 glycosylation 477:494 PD-1 glycosylation 477:494 It has previously been speculated that PD-1 glycosylation is involved in nivolumab recognition. 28165004 2 2 used used 254:257 arg2 Nivolumab 183:191 Nivolumab 183:191 Nivolumab 183:191 Nivolumab, a human monoclonal antibody targeting PD-1, has been widely used clinically since 2014. 28165004 2 2 used used 254:257 arg2 antibody 213:220 a human monoclonal antibody 194:220 a human monoclonal antibody targeting PD-1 194:235 Nivolumab, a human monoclonal antibody targeting PD-1, has been widely used clinically since 2014. 28165004 5 3 theme PD-1 622:625 arg1 N-glycosylation 627:641 PD-1 N-glycosylation 622:641 PD-1 N-glycosylation 622:641 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 5 4 theme N-glycosylation 627:641 arg1 effects 611:617 the effects 607:617 the effects of PD-1 N-glycosylation on the interactions with nivolumab 607:676 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 5 5 theme nivolumab 574:582 arg1 structure 561:569 the complex structure 549:569 the complex structure of nivolumab with PD-1 549:592 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 7 6 theme PD-L1 829:833 arg1 recognition 814:824 recognition 814:824 recognition of PD-L1 814:833 This loop is not involved in recognition of PD-L1 but dominates binding to nivolumab, whereas N-glycosylation is not involved in binding at all. 28165004 6 7 theme PD-1 779:782 arg1 domain 769:774 the IgV domain 761:774 the IgV domain of PD-1 761:782 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 6 7 theme PD-1 779:782 arg1 PD-1 779:782 PD-1 779:782 PD-1 779:782 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 6 8 theme Structural 679:688 arg1 analyses 705:712 Structural and functional analyses 679:712 Structural and functional analyses 679:712 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 1 9 theme hot-spot 156:163 arg1 research 146:153 a research 144:153 a research 'hot-spot' in recent years 144:180 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 1 9 theme hot-spot 156:163 arg1 immunotherapy 77:89 Cancer immunotherapy 70:89 Cancer immunotherapy by targeting of immune checkpoint molecules 70:133 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 3 10 theme nivolumab 316:324 arg1 mechanism 303:311 the binding mechanism 291:311 the binding mechanism of nivolumab to PD-1 291:332 However, the binding mechanism of nivolumab to PD-1 has not yet been shown, despite a recent report describing the complex structure of pembrolizumab/PD-1. 28165004 1 11 theme Cancer 70:75 arg1 research 146:153 a research 144:153 a research 'hot-spot' in recent years 144:180 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 1 11 theme Cancer 70:75 arg1 immunotherapy 77:89 Cancer immunotherapy 70:89 Cancer immunotherapy by targeting of immune checkpoint molecules 70:133 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 9 12 theme future 1047:1052 arg1 molecules 1065:1073 future inhibitory molecules 1047:1073 future inhibitory molecules targeting PD-1 1047:1088 These results provide the basis for the design of future inhibitory molecules targeting PD-1. 28165004 0 13 theme unexpected 3:12 arg1 loop 25:28 An unexpected N-terminal loop 0:28 An unexpected N-terminal loop in PD-1 0:36 An unexpected N-terminal loop in PD-1 dominates binding by nivolumab. 28165004 8 14 theme different 962:970 arg1 area 972:975 a completely different area 949:975 a completely different area than pembrolizumab 949:994 Nivolumab binds to a completely different area than pembrolizumab. 28165004 9 15 theme inhibitory 1054:1063 arg1 molecules 1065:1073 future inhibitory molecules 1047:1073 future inhibitory molecules targeting PD-1 1047:1088 These results provide the basis for the design of future inhibitory molecules targeting PD-1. 28165004 3 16 theme recent 368:373 arg1 report 375:380 a recent report 366:380 a recent report describing the complex structure of pembrolizumab/PD-1 366:435 However, the binding mechanism of nivolumab to PD-1 has not yet been shown, despite a recent report describing the complex structure of pembrolizumab/PD-1. 28165004 6 17 theme IgV 765:767 arg1 domain 769:774 the IgV domain 761:774 the IgV domain of PD-1 761:782 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 6 17 theme IgV 765:767 arg1 PD-1 779:782 PD-1 779:782 PD-1 779:782 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 9 18 theme molecules 1065:1073 arg1 design 1037:1042 the design 1033:1042 the design of future inhibitory molecules targeting PD-1 1033:1088 These results provide the basis for the design of future inhibitory molecules targeting PD-1. 28165004 1 19 theme recent 169:174 arg1 years 176:180 recent years 169:180 recent years 169:180 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 4 20 theme nivolumab 511:519 arg1 recognition 521:531 nivolumab recognition 511:531 nivolumab recognition 511:531 It has previously been speculated that PD-1 glycosylation is involved in nivolumab recognition. 28165004 5 21 theme complex 553:559 arg1 structure 561:569 the complex structure 549:569 the complex structure of nivolumab with PD-1 549:592 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 0 22 theme N-terminal 14:23 arg1 loop 25:28 An unexpected N-terminal loop 0:28 An unexpected N-terminal loop in PD-1 0:36 An unexpected N-terminal loop in PD-1 dominates binding by nivolumab. 28165004 6 23 theme N-terminal 737:746 arg1 loop 748:751 an N-terminal loop 734:751 an N-terminal loop outside the IgV domain of PD-1 734:782 Structural and functional analyses unexpectedly reveal an N-terminal loop outside the IgV domain of PD-1. 28165004 3 24 theme pembrolizumab/PD-1 418:435 arg1 structure 405:413 the complex structure 393:413 the complex structure of pembrolizumab/PD-1 393:435 However, the binding mechanism of nivolumab to PD-1 has not yet been shown, despite a recent report describing the complex structure of pembrolizumab/PD-1. 28165004 3 25 theme complex 397:403 arg1 structure 405:413 the complex structure 393:413 the complex structure of pembrolizumab/PD-1 393:435 However, the binding mechanism of nivolumab to PD-1 has not yet been shown, despite a recent report describing the complex structure of pembrolizumab/PD-1. 28165004 5 26 with interactions 650:661 arg1 nivolumab 668:676 nivolumab 668:676 nivolumab 668:676 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 5 27 with nivolumab 574:582 arg1 PD-1 589:592 PD-1 589:592 PD-1 589:592 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 2 28 theme monoclonal 202:211 arg1 antibody 213:220 a human monoclonal antibody 194:220 a human monoclonal antibody targeting PD-1 194:235 Nivolumab, a human monoclonal antibody targeting PD-1, has been widely used clinically since 2014. 28165004 2 28 theme monoclonal 202:211 arg1 Nivolumab 183:191 Nivolumab 183:191 Nivolumab 183:191 Nivolumab, a human monoclonal antibody targeting PD-1, has been widely used clinically since 2014. 28165004 2 29 theme human 196:200 arg1 antibody 213:220 a human monoclonal antibody 194:220 a human monoclonal antibody targeting PD-1 194:235 Nivolumab, a human monoclonal antibody targeting PD-1, has been widely used clinically since 2014. 28165004 2 29 theme human 196:200 arg1 Nivolumab 183:191 Nivolumab 183:191 Nivolumab 183:191 Nivolumab, a human monoclonal antibody targeting PD-1, has been widely used clinically since 2014. 28165004 1 30 theme immune 107:112 arg1 molecules 125:133 immune checkpoint molecules 107:133 immune checkpoint molecules 107:133 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 5 31 from effects 611:617 arg1 interactions 650:661 the interactions 646:661 the interactions with nivolumab 646:676 Here we report the complex structure of nivolumab with PD-1 and evaluate the effects of PD-1 N-glycosylation on the interactions with nivolumab. 28165004 1 32 from years 176:180 arg1 hot-spot 156:163 hot-spot 156:163 hot-spot 156:163 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 1 33 theme checkpoint 114:123 arg1 molecules 125:133 immune checkpoint molecules 107:133 immune checkpoint molecules 107:133 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 1 34 theme molecules 125:133 arg1 targeting 94:102 targeting 94:102 targeting of immune checkpoint molecules 94:133 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 28165004 7 35 dep involved 802:809 arg1 whereas 871:877 whereas 871:877 whereas 871:877 This loop is not involved in recognition of PD-L1 but dominates binding to nivolumab, whereas N-glycosylation is not involved in binding at all. 28165004 4 36 gly glycosylation 482:494 arg1 recognition 521:531 nivolumab recognition 511:531 nivolumab recognition 511:531 It has previously been speculated that PD-1 glycosylation is involved in nivolumab recognition. 28165004 3 37 theme binding 295:301 arg1 mechanism 303:311 the binding mechanism 291:311 the binding mechanism of nivolumab to PD-1 291:332 However, the binding mechanism of nivolumab to PD-1 has not yet been shown, despite a recent report describing the complex structure of pembrolizumab/PD-1. 28165004 0 38 from loop 25:28 arg1 PD-1 33:36 PD-1 33:36 PD-1 33:36 An unexpected N-terminal loop in PD-1 dominates binding by nivolumab. 28165004 1 39 from hot-spot 156:163 arg1 years 176:180 recent years 169:180 recent years 169:180 Cancer immunotherapy by targeting of immune checkpoint molecules has been a research 'hot-spot' in recent years. 3056714 8 0 theme proteinase 1173:1182 arg1 K 1184:1184 proteinase K 1173:1184 proteinase K 1173:1184 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 1 1 theme higher 226:231 arg1 activities 240:249 70-fold higher enzyme activities 218:249 70-fold higher enzyme activities of acid phosphatase 218:269 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 5 2 theme mannose-6-phosphate 804:822 arg1 receptors 824:832 mannose-6-phosphate receptors 804:832 mannose-6-phosphate receptors 804:832 Lack of mannose-6-phosphate residues suggested that transport does not involve mannose-6-phosphate receptors. 3056714 1 3 theme human 157:161 arg1 phosphatase 178:188 human lysosomal acid phosphatase 157:188 human lysosomal acid phosphatase (LAP) cDNA (CT29) 157:206 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 1 3 theme human 157:161 arg1 LAP 191:193 LAP 191:193 LAP 191:193 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 1 4 theme enzyme 233:238 arg1 activities 240:249 70-fold higher enzyme activities 218:249 70-fold higher enzyme activities of acid phosphatase 218:269 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 0 5 theme transfected 91:101 arg1 cells 123:127 transfected baby hamster kidney cells 91:127 transfected baby hamster kidney cells 91:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 8 6 theme CT29-derived 1063:1074 arg1 cRNA 1076:1079 CT29-derived cRNA 1063:1079 CT29-derived cRNA in the presence of microsomal membranes 1063:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 6 7 theme CT29-LAP 867:874 arg1 Part 835:838 Part 835:838 Part of the membrane-associated CT29-LAP 835:874 Part of the membrane-associated CT29-LAP was processed to a soluble form. 3056714 10 8 theme acids 1593:1597 arg1 tail 1576:1579 the cytosolic tail 1562:1579 the cytosolic tail of 18 amino acids 1562:1597 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 10 8 theme acids 1593:1597 arg1 acids 1593:1597 18 amino acids 1584:1597 18 amino acids 1584:1597 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 10 8 theme acids 1593:1597 arg1 domain 1551:1556 the transmembrane domain 1533:1556 the transmembrane domain 1533:1556 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 10 9 theme amino 1587:1591 arg1 acids 1593:1597 18 amino acids 1584:1597 18 amino acids 1584:1597 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 6 10 theme membrane-associated 847:865 arg1 CT29-LAP 867:874 the membrane-associated CT29-LAP 843:874 the membrane-associated CT29-LAP 843:874 Part of the membrane-associated CT29-LAP was processed to a soluble form. 3056714 3 11 theme partial 525:531 arg1 processing 533:542 partial processing 525:542 partial processing of the oligosaccharides to complex type structures 525:593 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 8 12 dep In 1039:1040 arg1 vitro 1042:1046 vitro 1042:1046 vitro 1042:1046 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 3 13 theme small 477:481 arg1 increase 483:490 a small increase 475:490 a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures 475:593 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 0 14 theme hamster 108:114 arg1 kidney 116:121 baby hamster kidney 103:121 transfected baby hamster kidney cells 91:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 10 15 theme cytosolic 1566:1574 arg1 tail 1576:1579 the cytosolic tail 1562:1579 the cytosolic tail of 18 amino acids 1562:1597 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 2 16 theme glycosylated 366:377 arg1 precursor 379:387 a heterogeneously glycosylated precursor 348:387 a heterogeneously glycosylated precursor that was tightly membrane associated 348:424 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 2 16 theme glycosylated 366:377 arg1 CT29-LAP 307:314 The CT29-LAP 303:314 The CT29-LAP 303:314 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 2 16 theme glycosylated 366:377 arg1 associated 415:424 associated 415:424 associated 415:424 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 1 17 theme lysosomal 163:171 arg1 phosphatase 178:188 human lysosomal acid phosphatase 157:188 human lysosomal acid phosphatase (LAP) cDNA (CT29) 157:206 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 1 17 theme lysosomal 163:171 arg1 LAP 191:193 LAP 191:193 LAP 191:193 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 0 18 theme baby 103:106 arg1 kidney 116:121 baby hamster kidney 103:121 transfected baby hamster kidney cells 91:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 8 19 theme cRNA 1076:1079 arg1 translation 1048:1058 In vitro translation 1039:1058 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes 1039:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 8 20 theme small 1199:1203 arg1 peptide 1205:1211 a small peptide 1197:1211 a small peptide of approximately 2 kd 1197:1233 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 1 21 theme acid 173:176 arg1 phosphatase 178:188 human lysosomal acid phosphatase 157:188 human lysosomal acid phosphatase (LAP) cDNA (CT29) 157:206 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 1 21 theme acid 173:176 arg1 LAP 191:193 LAP 191:193 LAP 191:193 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 0 22 theme lysosomal 6:14 arg1 phosphatase 21:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 0 22 theme lysosomal 6:14 arg1 protein 67:73 a transmembrane protein 51:73 a transmembrane protein to lysosomes in transfected baby hamster kidney cells 51:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 8 23 theme In 1039:1040 arg1 translation 1048:1058 In vitro translation 1039:1058 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes 1039:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 1 24 theme phosphatase 178:188 arg1 CT29 202:205 CT29 202:205 CT29 202:205 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 1 24 theme phosphatase 178:188 arg1 cDNA 196:199 human lysosomal acid phosphatase (LAP) cDNA 157:199 human lysosomal acid phosphatase (LAP) cDNA (CT29) 157:206 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 0 25 theme Human 0:4 arg1 phosphatase 21:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 0 25 theme Human 0:4 arg1 protein 67:73 a transmembrane protein 51:73 a transmembrane protein to lysosomes in transfected baby hamster kidney cells 51:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 1 26 theme acid 254:257 arg1 phosphatase 259:269 acid phosphatase 254:269 acid phosphatase 254:269 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 2 27 gly glycosylated 366:377 arg1 precursor 379:387 a heterogeneously glycosylated precursor 348:387 a heterogeneously glycosylated precursor that was tightly membrane associated 348:424 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 2 27 gly glycosylated 366:377 arg1 CT29-LAP 307:314 The CT29-LAP 303:314 The CT29-LAP 303:314 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 2 27 gly glycosylated 366:377 arg1 associated 415:424 associated 415:424 associated 415:424 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 3 28 from increase 483:490 arg1 kd 517:518 approximately 7 kd 501:518 approximately 7 kd 501:518 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 3 28 from increase 483:490 arg1 size 495:498 size 495:498 size (approximately 7 kd) 495:519 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 3 28 from increase 483:490 arg1 processing 533:542 partial processing 525:542 partial processing of the oligosaccharides to complex type structures 525:593 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 3 29 theme oligosaccharides 551:566 arg1 kd 517:518 approximately 7 kd 501:518 approximately 7 kd 501:518 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 3 29 theme oligosaccharides 551:566 arg1 processing 533:542 partial processing 525:542 partial processing of the oligosaccharides to complex type structures 525:593 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 3 29 theme oligosaccharides 551:566 arg1 size 495:498 size 495:498 size (approximately 7 kd) 495:519 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 4 30 theme immunoelectron 698:711 arg1 microscopy 713:722 immunoelectron microscopy 698:722 immunoelectron microscopy 698:722 CT29-LAP was transferred into lysosomes as shown by subcellular fractionation, immunofluorescence and immunoelectron microscopy. 3056714 1 31 theme phosphatase 259:269 arg1 activities 240:249 70-fold higher enzyme activities 218:249 70-fold higher enzyme activities of acid phosphatase 218:269 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 8 32 theme microsomal 1100:1109 arg1 membranes 1111:1119 microsomal membranes 1100:1119 microsomal membranes 1100:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 10 33 theme proteolytic 1466:1476 arg1 cleavage 1478:1485 proteolytic cleavage 1466:1485 proteolytic cleavage 1466:1485 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 0 34 theme acid 16:19 arg1 phosphatase 21:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 0 34 theme acid 16:19 arg1 protein 67:73 a transmembrane protein 51:73 a transmembrane protein to lysosomes in transfected baby hamster kidney cells 51:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 8 35 from translation 1048:1058 arg1 presence 1088:1095 the presence 1084:1095 the presence of microsomal membranes 1084:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 0 36 theme kidney 116:121 arg1 cells 123:127 transfected baby hamster kidney cells 91:127 transfected baby hamster kidney cells 91:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 2 37 theme BHK 335:337 arg1 cells 339:343 BHK cells 335:343 BHK cells 335:343 The CT29-LAP was synthesized in BHK cells as a heterogeneously glycosylated precursor that was tightly membrane associated. 3056714 9 38 with combination 1239:1249 arg1 data 1269:1272 the sequence data 1256:1272 the sequence data available for LAP 1256:1290 In combination with the sequence data available for LAP, these observations suggest that CT29-LAP is synthesized and transported to lysosomes as a transmembrane protein. 3056714 8 39 theme kd 1232:1233 arg1 peptide 1205:1211 a small peptide 1197:1211 a small peptide of approximately 2 kd 1197:1233 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 1 40 theme non-transfected 276:290 arg1 cells 296:300 non-transfected BHK cells 276:300 non-transfected BHK cells 276:300 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 3 41 theme type 579:582 arg1 structures 584:593 complex type structures 571:593 complex type structures 571:593 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 4 42 theme subcellular 648:658 arg1 fractionation 660:672 subcellular fractionation 648:672 subcellular fractionation 648:672 CT29-LAP was transferred into lysosomes as shown by subcellular fractionation, immunofluorescence and immunoelectron microscopy. 3056714 1 43 theme BHK 292:294 arg1 cells 296:300 non-transfected BHK cells 276:300 non-transfected BHK cells 276:300 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 5 44 theme mannose-6-phosphate 733:751 arg1 residues 753:760 mannose-6-phosphate residues 733:760 mannose-6-phosphate residues 733:760 Lack of mannose-6-phosphate residues suggested that transport does not involve mannose-6-phosphate receptors. 3056714 0 45 from protein 67:73 arg1 cells 123:127 transfected baby hamster kidney cells 91:127 transfected baby hamster kidney cells 91:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 5 46 theme residues 753:760 arg1 Lack 725:728 Lack 725:728 Lack of mannose-6-phosphate residues 725:760 Lack of mannose-6-phosphate residues suggested that transport does not involve mannose-6-phosphate receptors. 3056714 8 47 theme CT29-LAP 1131:1138 arg1 precursor 1140:1148 a CT29-LAP precursor 1129:1148 a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd 1129:1233 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 9 48 theme sequence 1260:1267 arg1 data 1269:1272 the sequence data 1256:1272 the sequence data available for LAP 1256:1290 In combination with the sequence data available for LAP, these observations suggest that CT29-LAP is synthesized and transported to lysosomes as a transmembrane protein. 3056714 8 49 theme membranes 1111:1119 arg1 presence 1088:1095 the presence 1084:1095 the presence of microsomal membranes 1084:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 9 50 theme transmembrane 1383:1395 arg1 CT29-LAP 1325:1332 CT29-LAP 1325:1332 CT29-LAP 1325:1332 In combination with the sequence data available for LAP, these observations suggest that CT29-LAP is synthesized and transported to lysosomes as a transmembrane protein. 3056714 9 50 theme transmembrane 1383:1395 arg1 protein 1397:1403 a transmembrane protein 1381:1403 a transmembrane protein 1381:1403 In combination with the sequence data available for LAP, these observations suggest that CT29-LAP is synthesized and transported to lysosomes as a transmembrane protein. 3056714 10 51 theme transmembrane 1537:1549 arg1 acids 1593:1597 18 amino acids 1584:1597 18 amino acids 1584:1597 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 10 51 theme transmembrane 1537:1549 arg1 domain 1551:1556 the transmembrane domain 1533:1556 the transmembrane domain 1533:1556 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 6 52 theme soluble 895:901 arg1 form 903:906 a soluble form 893:906 a soluble form 893:906 Part of the membrane-associated CT29-LAP was processed to a soluble form. 3056714 3 53 theme complex 571:577 arg1 structures 584:593 complex type structures 571:593 complex type structures 571:593 Transfer to the trans-Golgi was associated with a small increase in size (approximately 7 kd) and partial processing of the oligosaccharides to complex type structures. 3056714 8 54 link CT29-derived 1063:1074 arg1 cRNA 1076:1079 CT29-derived cRNA 1063:1079 CT29-derived cRNA in the presence of microsomal membranes 1063:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 8 55 from cRNA 1076:1079 arg1 presence 1088:1095 the presence 1084:1095 the presence of microsomal membranes 1084:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 8 56 from presence 1088:1095 arg1 translation 1048:1058 In vitro translation 1039:1058 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes 1039:1119 In vitro translation of CT29-derived cRNA in the presence of microsomal membranes yielded a CT29-LAP precursor that is protected from proteinase K except for a small peptide of approximately 2 kd. 3056714 9 57 theme available 1274:1282 arg1 data 1269:1272 the sequence data 1256:1272 the sequence data available for LAP 1256:1290 In combination with the sequence data available for LAP, these observations suggest that CT29-LAP is synthesized and transported to lysosomes as a transmembrane protein. 3056714 7 58 theme polypeptide 1018:1028 arg1 size 1006:1009 the size 1002:1009 the size of the polypeptide 1002:1028 The mechanism that converts CT29-LAP into a soluble form was sensitive to NH4Cl, and reduced the size of the polypeptide by 7 kd. 3056714 1 59 theme BHK 130:132 arg1 cells 134:138 BHK cells 130:138 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) 130:206 BHK cells transfected with human lysosomal acid phosphatase (LAP) cDNA (CT29) expressed 70-fold higher enzyme activities of acid phosphatase than non-transfected BHK cells. 3056714 10 60 attach released 1436:1443 arg1 membrane 1454:1461 the membrane 1450:1461 the membrane 1450:1461 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 10 60 attach released 1436:1443 arg2 CT29-LAP 1424:1431 CT29-LAP 1424:1431 CT29-LAP 1424:1431 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 10 61 theme C-terminal 1504:1513 arg1 peptide 1515:1521 a C-terminal peptide 1502:1521 a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids 1502:1597 In the lysosomes, CT29-LAP is released from the membrane by proteolytic cleavage, which removes a C-terminal peptide including the transmembrane domain and the cytosolic tail of 18 amino acids. 3056714 7 62 theme soluble 953:959 arg1 form 961:964 a soluble form 951:964 a soluble form 951:964 The mechanism that converts CT29-LAP into a soluble form was sensitive to NH4Cl, and reduced the size of the polypeptide by 7 kd. 3056714 0 63 theme transmembrane 53:65 arg1 phosphatase 21:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase 0:31 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 3056714 0 63 theme transmembrane 53:65 arg1 protein 67:73 a transmembrane protein 51:73 a transmembrane protein to lysosomes in transfected baby hamster kidney cells 51:127 Human lysosomal acid phosphatase is transported as a transmembrane protein to lysosomes in transfected baby hamster kidney cells. 25760607 0 0 theme NKR-P1 57:62 arg1 LLT1 33:36 human LLT1 27:36 human LLT1 27:36 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 0 0 theme NKR-P1 57:62 arg1 ligand 41:46 a ligand 39:46 a ligand of human NKR-P1 39:62 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 0 1 from structures 13:22 arg1 states 109:114 oligomerization states 93:114 oligomerization states 93:114 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 0 1 from structures 13:22 arg1 glycosylation 75:87 varied glycosylation 68:87 varied glycosylation 68:87 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 3 2 theme N-acetylglucosamine 451:469 arg1 unit 471:474 the first N-acetylglucosamine unit 441:474 the first N-acetylglucosamine unit 441:474 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 5 3 theme outer 699:703 arg1 part 705:708 an outer part 696:708 an outer part of the long loop region 696:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 4 4 theme dimeric 551:557 arg1 form 559:562 The dimeric form 547:562 The dimeric form 547:562 The dimeric form follows the classical dimerization mode of human CD69. 25760607 0 5 theme human 51:55 arg1 NKR-P1 57:62 human NKR-P1 51:62 human NKR-P1 51:62 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 5 6 theme loop 722:725 arg1 region 727:732 the long loop region 713:732 the long loop region 713:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 5 7 theme monomeric 623:631 arg1 form 633:636 The monomeric form 619:636 The monomeric form 619:636 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 6 8 theme LLT1 763:766 arg1 hexamer 739:745 The hexamer 735:745 The hexamer of glycosylated LLT1 735:766 The hexamer of glycosylated LLT1 consists of three classical dimers. 25760607 1 9 theme C-type 192:197 arg1 NKR-P1 162:167 NKR-P1 162:167 NKR-P1 (CD161, gene KLRB1) 162:187 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 9 theme C-type 192:197 arg1 receptor 211:218 a C-type lectin-like receptor 190:218 a C-type lectin-like receptor of natural killer cells 190:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 5 10 theme position 684:691 arg1 exception 667:675 the exception 663:675 the exception of the position of an outer part of the long loop region 663:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 6 11 theme glycosylated 750:761 arg1 LLT1 763:766 glycosylated LLT1 750:766 glycosylated LLT1 750:766 The hexamer of glycosylated LLT1 consists of three classical dimers. 25760607 1 12 theme lectin-like 199:209 arg1 NKR-P1 162:167 NKR-P1 162:167 NKR-P1 (CD161, gene KLRB1) 162:187 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 12 theme lectin-like 199:209 arg1 receptor 211:218 a C-type lectin-like receptor 190:218 a C-type lectin-like receptor of natural killer cells 190:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 7 13 theme hexameric 808:816 arg1 packing 818:824 The hexameric packing 804:824 The hexameric packing 804:824 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 1 14 theme C-type 133:138 arg1 ligand 152:157 a C-type lectin-like ligand 131:157 a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells 131:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 14 theme C-type 133:138 arg1 LLT1 123:126 Human LLT1 117:126 Human LLT1 117:126 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 0 15 theme varied 68:73 arg1 glycosylation 75:87 varied glycosylation 68:87 varied glycosylation 68:87 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 3 16 theme LLT1 354:357 arg1 structures 340:349 Four structures 335:349 Four structures of LLT1 under various conditions 335:382 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 3 17 from deglycosylated 420:433 arg1 hexameric 493:501 hexameric 493:501 hexameric 493:501 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 3 17 from deglycosylated 420:433 arg1 forms 483:487 two forms 479:487 two forms 479:487 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 1 18 theme lectin-like 140:150 arg1 ligand 152:157 a C-type lectin-like ligand 131:157 a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells 131:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 18 theme lectin-like 140:150 arg1 LLT1 123:126 Human LLT1 117:126 Human LLT1 117:126 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 0 19 theme crystal 5:11 arg1 structures 13:22 Four crystal structures 0:22 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 0:115 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 5 20 theme long 717:720 arg1 region 727:732 the long loop region 713:732 the long loop region 713:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 7 21 theme proteins 892:899 arg1 interaction 858:868 interaction 858:868 interaction of C-type lectin-like proteins 858:899 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 3 22 theme various 365:371 arg1 conditions 373:382 various conditions 365:382 various conditions 365:382 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 2 23 theme X-ray 251:255 arg1 diffraction 257:267 X-ray diffraction 251:267 X-ray diffraction 251:267 Using X-ray diffraction, the first experimental structures of human LLT1 were determined. 25760607 1 24 theme natural 223:229 arg1 cells 238:242 natural killer cells 223:242 natural killer cells 223:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 0 25 theme oligomerization 93:107 arg1 states 109:114 oligomerization states 93:114 oligomerization states 93:114 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 7 26 theme C-type 873:878 arg1 proteins 892:899 C-type lectin-like proteins 873:899 C-type lectin-like proteins 873:899 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 3 27 theme homogeneous 508:518 arg1 glycosylation 532:544 homogeneous GlcNAc2Man5 glycosylation 508:544 homogeneous GlcNAc2Man5 glycosylation 508:544 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 7 28 theme glycosylated 908:919 arg1 form 921:924 the glycosylated form 904:924 the glycosylated form 904:924 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 1 29 theme NKR-P1 162:167 arg1 ligand 152:157 a C-type lectin-like ligand 131:157 a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells 131:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 29 theme NKR-P1 162:167 arg1 LLT1 123:126 Human LLT1 117:126 Human LLT1 117:126 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 7 30 from mode 850:853 arg1 form 921:924 the glycosylated form 904:924 the glycosylated form 904:924 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 1 31 theme killer 231:236 arg1 cells 238:242 natural killer cells 223:242 natural killer cells 223:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 4 32 theme classical 576:584 arg1 mode 599:602 the classical dimerization mode 572:602 the classical dimerization mode of human CD69 572:616 The dimeric form follows the classical dimerization mode of human CD69. 25760607 3 33 theme GlcNAc2Man5 520:530 arg1 glycosylation 532:544 homogeneous GlcNAc2Man5 glycosylation 508:544 homogeneous GlcNAc2Man5 glycosylation 508:544 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 1 34 theme Human 117:121 arg1 ligand 152:157 a C-type lectin-like ligand 131:157 a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells 131:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 34 theme Human 117:121 arg1 LLT1 123:126 Human LLT1 117:126 Human LLT1 117:126 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 0 35 theme LLT1 33:36 arg1 structures 13:22 Four crystal structures 0:22 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 0:115 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 1 36 theme cells 238:242 arg1 NKR-P1 162:167 NKR-P1 162:167 NKR-P1 (CD161, gene KLRB1) 162:187 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 36 theme cells 238:242 arg1 receptor 211:218 a C-type lectin-like receptor 190:218 a C-type lectin-like receptor of natural killer cells 190:242 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 0 37 theme human 27:31 arg1 LLT1 33:36 human LLT1 27:36 human LLT1 27:36 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 0 37 theme human 27:31 arg1 ligand 41:46 a ligand 39:46 a ligand of human NKR-P1 39:62 Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states. 25760607 6 38 gly glycosylated 750:761 arg1 LLT1 763:766 glycosylated LLT1 750:766 glycosylated LLT1 750:766 The hexamer of glycosylated LLT1 consists of three classical dimers. 25760607 5 39 theme region 727:732 arg1 part 705:708 an outer part 696:708 an outer part of the long loop region 696:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 3 40 theme monomeric 401:409 arg1 deglycosylated 420:433 monomeric, dimeric deglycosylated 401:433 monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation 401:544 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 3 41 with hexameric 493:501 arg1 glycosylation 532:544 homogeneous GlcNAc2Man5 glycosylation 508:544 homogeneous GlcNAc2Man5 glycosylation 508:544 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 7 42 gly glycosylated 908:919 arg1 form 921:924 the glycosylated form 904:924 the glycosylated form 904:924 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 2 43 theme experimental 280:291 arg1 structures 293:302 the first experimental structures 270:302 the first experimental structures of human LLT1 270:316 Using X-ray diffraction, the first experimental structures of human LLT1 were determined. 25760607 5 44 theme same 648:651 arg1 fold 653:656 the same fold 644:656 the same fold with the exception of the position of an outer part of the long loop region 644:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 6 45 theme classical 786:794 arg1 dimers 796:801 three classical dimers 780:801 three classical dimers 780:801 The hexamer of glycosylated LLT1 consists of three classical dimers. 25760607 3 46 dep determined 389:398 arg1 deglycosylated 420:433 monomeric, dimeric deglycosylated 401:433 monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation 401:544 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 2 47 theme first 274:278 arg1 structures 293:302 the first experimental structures 270:302 the first experimental structures of human LLT1 270:316 Using X-ray diffraction, the first experimental structures of human LLT1 were determined. 25760607 7 48 theme lectin-like 880:890 arg1 proteins 892:899 C-type lectin-like proteins 873:899 C-type lectin-like proteins 873:899 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 7 49 theme interaction 858:868 arg1 mode 850:853 a possible mode 839:853 a possible mode of interaction of C-type lectin-like proteins in the glycosylated form 839:924 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 1 50 theme gene 177:180 arg1 CD161 170:174 CD161 170:174 CD161 170:174 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 50 theme gene 177:180 arg1 KLRB1 182:186 gene KLRB1 177:186 gene KLRB1 177:186 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 3 51 dep monomeric 401:409 arg1 dimeric 412:418 dimeric 412:418 dimeric 412:418 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 5 52 with fold 653:656 arg1 exception 667:675 the exception 663:675 the exception of the position of an outer part of the long loop region 663:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 7 53 theme possible 841:848 arg1 mode 850:853 a possible mode 839:853 a possible mode of interaction of C-type lectin-like proteins in the glycosylated form 839:924 The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form. 25760607 5 54 theme part 705:708 arg1 position 684:691 the position 680:691 the position of an outer part of the long loop region 680:732 The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. 25760607 4 55 theme CD69 613:616 arg1 mode 599:602 the classical dimerization mode 572:602 the classical dimerization mode of human CD69 572:616 The dimeric form follows the classical dimerization mode of human CD69. 25760607 1 56 dep NKR-P1 162:167 arg1 CD161 170:174 CD161 170:174 CD161 170:174 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 1 56 dep NKR-P1 162:167 arg1 KLRB1 182:186 gene KLRB1 177:186 gene KLRB1 177:186 Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. 25760607 4 57 theme human 607:611 arg1 CD69 613:616 human CD69 607:616 human CD69 607:616 The dimeric form follows the classical dimerization mode of human CD69. 25760607 2 58 theme LLT1 313:316 arg1 structures 293:302 the first experimental structures 270:302 the first experimental structures of human LLT1 270:316 Using X-ray diffraction, the first experimental structures of human LLT1 were determined. 25760607 2 59 theme human 307:311 arg1 LLT1 313:316 human LLT1 307:316 human LLT1 307:316 Using X-ray diffraction, the first experimental structures of human LLT1 were determined. 25760607 3 60 theme first 445:449 arg1 unit 471:474 the first N-acetylglucosamine unit 441:474 the first N-acetylglucosamine unit 441:474 Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. 25760607 4 61 theme dimerization 586:597 arg1 mode 599:602 the classical dimerization mode 572:602 the classical dimerization mode of human CD69 572:616 The dimeric form follows the classical dimerization mode of human CD69. 25765764 5 0 theme LRIG1 739:743 arg1 domains 745:751 the LRIG1 domains 735:751 the LRIG1 domains 735:751 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 1 1 theme insect 311:316 arg1 cells 318:322 insect cells 311:322 insect cells 311:322 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 7 2 theme LRIG1-3Ig 1067:1075 arg1 domains 1077:1083 the LRIG1-3Ig domains 1063:1083 the LRIG1-3Ig domains 1063:1083 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 5 3 dep detect 713:718 arg1 either 811:816 either 811:816 either 811:816 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 5 3 dep detect 713:718 arg1 expressed 876:884 expressed 876:884 was expressed on the cell surface 872:904 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 1 4 theme extracellular 140:152 arg1 LRIG1-ECD 129:137 the human LRIG1-ECD 119:137 the human LRIG1-ECD (extracellular domain) 119:160 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 4 theme extracellular 140:152 arg1 domain 154:159 extracellular domain 140:159 extracellular domain 140:159 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 5 5 theme domains 745:751 arg1 binding 724:730 any binding 720:730 any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR) 720:808 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 1 6 from vectors 300:306 arg1 cells 318:322 insect cells 311:322 insect cells 311:322 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 7 theme immunoglobulin-like 222:240 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 7 theme immunoglobulin-like 222:240 arg1 domain 243:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 7 8 theme cell-surface 1182:1193 arg1 EGFR 1195:1198 cell-surface EGFR 1182:1198 cell-surface EGFR 1182:1198 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 4 9 theme LRIG1-LRR 574:582 arg1 domain 584:589 The LRIG1-LRR domain 570:589 The LRIG1-LRR domain 570:589 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 4 9 theme LRIG1-LRR 574:582 arg1 monomers 626:633 monomers 626:633 monomers in solution 626:645 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 4 9 theme LRIG1-LRR 574:582 arg1 fragment 613:620 the LRIG1-LRR-1Ig fragment 595:620 the LRIG1-LRR-1Ig fragment 595:620 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 7 10 theme soluble 1041:1047 arg1 LRIG1-LRR 1049:1057 the soluble LRIG1-LRR 1037:1057 the soluble LRIG1-LRR 1037:1057 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 1 11 dep fragments 106:114 arg1 domain 199:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 11 dep fragments 106:114 arg1 repeat 191:196 leucine-rich repeat 178:196 leucine-rich repeat 178:196 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 11 dep fragments 106:114 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 11 dep fragments 106:114 arg1 fragment 273:280 the LRIG1-LRR-1Ig fragment 255:280 the LRIG1-LRR-1Ig fragment 255:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 11 dep fragments 106:114 arg1 domain 243:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 7 12 theme ligand-stimulated 1150:1166 arg1 activation 1168:1177 ligand-stimulated activation 1150:1177 ligand-stimulated activation of cell-surface EGFR 1150:1198 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 2 13 theme three-dimensional 403:419 arg1 structures 421:430 the three-dimensional structures 399:430 the three-dimensional structures at 2.3Å resolution 399:449 The two LRIG1 domains crystallised so that we have been able to determine the three-dimensional structures at 2.3Å resolution. 25765764 0 14 theme extracellular 6:18 arg1 domain 20:25 LRIG1 extracellular domain 0:25 LRIG1 extracellular domain 0:25 LRIG1 extracellular domain: structure and function analysis. 25765764 2 15 from resolution 440:449 arg1 structures 421:430 the three-dimensional structures 399:430 the three-dimensional structures at 2.3Å resolution 399:449 The two LRIG1 domains crystallised so that we have been able to determine the three-dimensional structures at 2.3Å resolution. 25765764 7 16 theme EGFR 1195:1198 arg1 activation 1168:1177 ligand-stimulated activation 1150:1177 ligand-stimulated activation of cell-surface EGFR 1150:1198 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 3 17 theme LINGO-1 551:557 arg1 structure 559:567 the LINGO-1 structure 547:567 the LINGO-1 structure 547:567 We developed a three-dimensional structure for the LRIG1-ECD using homology modelling based on the LINGO-1 structure. 25765764 7 18 theme HEK293 1127:1132 arg1 cells 1134:1138 HEK293 cells 1127:1138 HEK293 cells 1127:1138 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 4 19 theme LRIG1-3Ig 660:668 arg1 domain 670:675 the LRIG1-3Ig domain 656:675 the LRIG1-3Ig domain 656:675 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 4 19 theme LRIG1-3Ig 660:668 arg1 dimeric 691:697 dimeric 691:697 dimeric 691:697 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 0 20 theme LRIG1 0:4 arg1 domain 20:25 LRIG1 extracellular domain 0:25 LRIG1 extracellular domain 0:25 LRIG1 extracellular domain: structure and function analysis. 25765764 6 21 theme LRIG1-LRR-1Ig 923:935 arg1 fragment 937:944 The FLAG-tagged LRIG1-LRR-1Ig fragment 907:944 The FLAG-tagged LRIG1-LRR-1Ig fragment 907:944 The FLAG-tagged LRIG1-LRR-1Ig fragment binds weakly to colon cancer cells regardless of the presence of EGFRs. 25765764 1 22 theme LRIG1-LRR 167:175 arg1 domain 199:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 22 theme LRIG1-LRR 167:175 arg1 repeat 191:196 leucine-rich repeat 178:196 leucine-rich repeat 178:196 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 22 theme LRIG1-LRR 167:175 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 5 23 theme EGF 790:792 arg1 EGFR 804:807 EGFR 804:807 EGFR 804:807 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 5 23 theme EGF 790:792 arg1 receptor 794:801 EGF receptor 790:801 the EGF receptor (EGFR) 786:808 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 6 24 theme FLAG-tagged 911:921 arg1 fragment 937:944 The FLAG-tagged LRIG1-LRR-1Ig fragment 907:944 The FLAG-tagged LRIG1-LRR-1Ig fragment 907:944 The FLAG-tagged LRIG1-LRR-1Ig fragment binds weakly to colon cancer cells regardless of the presence of EGFRs. 25765764 5 25 theme cell 893:896 arg1 surface 898:904 the cell surface 889:904 the cell surface 889:904 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 1 26 theme soluble 98:104 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 26 theme soluble 98:104 arg1 domain 199:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 26 theme soluble 98:104 arg1 fragment 273:280 the LRIG1-LRR-1Ig fragment 255:280 the LRIG1-LRR-1Ig fragment 255:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 26 theme soluble 98:104 arg1 domain 243:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 4 27 from monomers 626:633 arg1 solution 638:645 solution 638:645 solution 638:645 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 1 28 theme leucine-rich 178:189 arg1 domain 199:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 28 theme leucine-rich 178:189 arg1 repeat 191:196 leucine-rich repeat 178:196 leucine-rich repeat 178:196 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 7 29 theme full-length 1093:1103 arg1 LRIG1 1105:1109 the full-length LRIG1 1089:1109 the full-length LRIG1 co-expressed in HEK293 cells 1089:1138 Similarly, neither the soluble LRIG1-LRR nor the LRIG1-3Ig domains nor the full-length LRIG1 co-expressed in HEK293 cells inhibited ligand-stimulated activation of cell-surface EGFR. 25765764 1 30 theme LRIG1-LRR-1Ig 259:271 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 30 theme LRIG1-LRR-1Ig 259:271 arg1 fragment 273:280 the LRIG1-LRR-1Ig fragment 255:280 the LRIG1-LRR-1Ig fragment 255:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 6 31 theme EGFRs 1011:1015 arg1 presence 999:1006 the presence 995:1006 the presence of EGFRs 995:1015 The FLAG-tagged LRIG1-LRR-1Ig fragment binds weakly to colon cancer cells regardless of the presence of EGFRs. 25765764 6 32 theme colon 962:966 arg1 cells 975:979 colon cancer cells 962:979 colon cancer cells 962:979 The FLAG-tagged LRIG1-LRR-1Ig fragment binds weakly to colon cancer cells regardless of the presence of EGFRs. 25765764 2 33 theme 2.3Å 435:438 arg1 resolution 440:449 2.3Å resolution 435:449 2.3Å resolution 435:449 The two LRIG1 domains crystallised so that we have been able to determine the three-dimensional structures at 2.3Å resolution. 25765764 5 34 theme biosensor 836:844 arg1 analysis 846:853 biosensor analysis 836:853 biosensor analysis 836:853 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 5 35 dep either 811:816 arg1 solution 821:828 solution 821:828 solution using biosensor analysis 821:853 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 5 36 theme fragment 774:781 arg1 binding 724:730 any binding 720:730 any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR) 720:808 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 1 37 theme baculovirus 288:298 arg1 vectors 300:306 baculovirus vectors 288:306 baculovirus vectors in insect cells 288:322 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 0 38 dep structure 28:36 arg1 domain 20:25 LRIG1 extracellular domain 0:25 LRIG1 extracellular domain 0:25 LRIG1 extracellular domain: structure and function analysis. 25765764 0 38 dep structure 28:36 arg1 analysis 51:58 analysis 51:58 analysis 51:58 LRIG1 extracellular domain: structure and function analysis. 25765764 3 39 theme three-dimensional 467:483 arg1 structure 485:493 a three-dimensional structure 465:493 a three-dimensional structure for the LRIG1-ECD using homology modelling based on the LINGO-1 structure 465:567 We developed a three-dimensional structure for the LRIG1-ECD using homology modelling based on the LINGO-1 structure. 25765764 3 40 theme homology 519:526 arg1 modelling 528:536 homology modelling 519:536 homology modelling based on the LINGO-1 structure 519:567 We developed a three-dimensional structure for the LRIG1-ECD using homology modelling based on the LINGO-1 structure. 25765764 1 41 theme human 123:127 arg1 LRIG1-ECD 129:137 the human LRIG1-ECD 119:137 the human LRIG1-ECD (extracellular domain) 119:160 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 41 theme human 123:127 arg1 domain 154:159 extracellular domain 140:159 extracellular domain 140:159 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 42 theme LRIG1-ECD 129:137 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 42 theme LRIG1-ECD 129:137 arg1 domain 199:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 the LRIG1-LRR (leucine-rich repeat) domain 163:204 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 42 theme LRIG1-ECD 129:137 arg1 fragment 273:280 the LRIG1-LRR-1Ig fragment 255:280 the LRIG1-LRR-1Ig fragment 255:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 42 theme LRIG1-ECD 129:137 arg1 domain 243:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 6 43 theme cancer 968:973 arg1 cells 975:979 colon cancer cells 962:979 colon cancer cells 962:979 The FLAG-tagged LRIG1-LRR-1Ig fragment binds weakly to colon cancer cells regardless of the presence of EGFRs. 25765764 5 44 theme LRIG1-LRR-1Ig 760:772 arg1 fragment 774:781 the LRIG1-LRR-1Ig fragment 756:781 the LRIG1-LRR-1Ig fragment 756:781 We could not detect any binding of the LRIG1 domains or the LRIG1-LRR-1Ig fragment to the EGF receptor (EGFR), either in solution using biosensor analysis or when the EGFR was expressed on the cell surface. 25765764 4 45 theme LRIG1-LRR-1Ig 599:611 arg1 domain 584:589 The LRIG1-LRR domain 570:589 The LRIG1-LRR domain 570:589 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 4 45 theme LRIG1-LRR-1Ig 599:611 arg1 monomers 626:633 monomers 626:633 monomers in solution 626:645 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 4 45 theme LRIG1-LRR-1Ig 599:611 arg1 fragment 613:620 the LRIG1-LRR-1Ig fragment 595:620 the LRIG1-LRR-1Ig fragment 595:620 The LRIG1-LRR domain and the LRIG1-LRR-1Ig fragment are monomers in solution, whereas the LRIG1-3Ig domain appears to be dimeric. 25765764 2 46 theme LRIG1 333:337 arg1 domains 339:345 The two LRIG1 domains 325:345 The two LRIG1 domains 325:345 The two LRIG1 domains crystallised so that we have been able to determine the three-dimensional structures at 2.3Å resolution. 25765764 1 47 theme LRIG1-3Ig 211:219 arg1 fragments 106:114 three soluble fragments 92:114 three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment 92:280 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 25765764 1 47 theme LRIG1-3Ig 211:219 arg1 domain 243:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 the LRIG1-3Ig (immunoglobulin-like) domain 207:248 We have expressed and purified three soluble fragments of the human LRIG1-ECD (extracellular domain): the LRIG1-LRR (leucine-rich repeat) domain, the LRIG1-3Ig (immunoglobulin-like) domain, and the LRIG1-LRR-1Ig fragment using baculovirus vectors in insect cells. 27991905 2 0 theme closed 294:299 arg1 conformation 301:312 a closed conformation 292:312 a closed conformation 292:312 Here we present the structure of human PC2 in a closed conformation, solved by electron cryomicroscopy at 4.2-Å resolution. 27991905 4 1 theme pore 769:772 arg1 filter 774:779 the pore filter 765:779 the pore filter 765:779 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 3 2 theme TOP 458:460 arg1 domain 463:468 polycystins' (TOP) domain 444:468 polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure 444:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 2 3 from structure 266:274 arg1 conformation 301:312 a closed conformation 292:312 a closed conformation 292:312 Here we present the structure of human PC2 in a closed conformation, solved by electron cryomicroscopy at 4.2-Å resolution. 27991905 5 4 theme channel-like 908:919 arg1 region 921:926 the homologous channel-like region 893:926 the homologous channel-like region of PC1 893:933 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 4 5 theme upper 742:746 arg1 vestibule 748:756 an upper vestibule 739:756 an upper vestibule 739:756 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 6 6 theme ample 1084:1088 arg1 scope 1090:1094 ample scope 1084:1094 ample scope for regulation through physical and chemical stimuli 1084:1147 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27991905 5 7 theme variants 994:1001 arg1 cluster 957:963 a cluster 955:963 a cluster of ADPKD-associated missense variants 955:1001 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 4 8 theme voltage-sensor-like 614:632 arg1 VSLD 642:645 VSLD 642:645 VSLD 642:645 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 4 8 theme voltage-sensor-like 614:632 arg1 domain 634:639 the voltage-sensor-like domain 610:639 the voltage-sensor-like domain (VSLD) 610:646 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 5 9 theme TOP-domain 831:840 arg1 fold 842:845 The TOP-domain fold 827:845 The TOP-domain fold 827:845 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 4 10 theme endoplasmic 673:683 arg1 reticulum 685:693 the channel's endoplasmic reticulum lumen or extracellular surface 659:724 reticulum 685:693 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 2 11 theme electron 325:332 arg1 cryomicroscopy 334:347 electron cryomicroscopy 325:347 electron cryomicroscopy 325:347 Here we present the structure of human PC2 in a closed conformation, solved by electron cryomicroscopy at 4.2-Å resolution. 27991905 1 12 theme unknown 226:232 arg1 mechanisms 234:243 unknown mechanisms 226:243 unknown mechanisms 226:243 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 13 dep polycystin-2 125:136 arg1 TRPP1 152:156 TRPP1 152:156 TRPP1 152:156 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 13 dep polycystin-2 125:136 arg1 PKD2 144:147 PKD2 144:147 PKD2 144:147 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 13 dep polycystin-2 125:136 arg1 PC2 139:141 PC2 139:141 PC2 139:141 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 0 14 theme polycystic 17:26 arg1 disease 35:41 the polycystic kidney disease 13:41 the polycystic kidney disease 13:41 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 6 15 theme physical 1119:1126 arg1 stimuli 1141:1147 physical and chemical stimuli 1119:1147 physical and chemical stimuli 1119:1147 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27991905 5 16 theme PC1 931:933 arg1 region 921:926 the homologous channel-like region 893:926 the homologous channel-like region of PC1 893:933 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 3 17 theme classic 500:506 arg1 channel 543:549 a classic transient receptor potential (TRP) channel 498:549 a classic transient receptor potential (TRP) channel structure 498:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 5 18 theme cluster 957:963 arg1 site 947:950 the site 943:950 the site of a cluster of ADPKD-associated missense variants 943:1001 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 5 18 theme cluster 957:963 arg1 cluster 957:963 a cluster 955:963 a cluster of ADPKD-associated missense variants 955:1001 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 0 19 dep Polycystin-2 55:66 arg1 Structure 0:8 Structure 0:8 Structure of the polycystic kidney disease 0:41 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 3 20 theme tetragonal 421:430 arg1 opening 432:438 a novel polycystin-specific 'tetragonal opening 392:438 a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure 392:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 0 21 theme disease 35:41 arg1 Structure 0:8 Structure 0:8 Structure of the polycystic kidney disease 0:41 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 6 22 theme TOP-domain 1033:1042 arg1 pore 1058:1061 the pore 1054:1061 the pore 1054:1061 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27991905 6 22 theme TOP-domain 1033:1042 arg1 VSLD 1071:1074 the VSLD 1067:1074 the VSLD 1067:1074 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27991905 6 22 theme TOP-domain 1033:1042 arg1 subunits 1044:1051 the TOP-domain subunits 1029:1051 the TOP-domain subunits 1029:1051 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27991905 3 23 theme novel 394:398 arg1 opening 432:438 a novel polycystin-specific 'tetragonal opening 392:438 a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure 392:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 3 24 theme potential 527:535 arg1 channel 543:549 a classic transient receptor potential (TRP) channel 498:549 a classic transient receptor potential (TRP) channel structure 498:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 2 25 theme 4.2-Å 352:356 arg1 resolution 358:367 4.2-Å resolution 352:367 4.2-Å resolution 352:367 Here we present the structure of human PC2 in a closed conformation, solved by electron cryomicroscopy at 4.2-Å resolution. 27991905 0 26 theme kidney 28:33 arg1 disease 35:41 the polycystic kidney disease 13:41 the polycystic kidney disease 13:41 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 5 27 theme ADPKD-associated 968:983 arg1 variants 994:1001 ADPKD-associated missense variants 968:1001 ADPKD-associated missense variants 968:1001 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 6 28 theme chemical 1132:1139 arg1 stimuli 1141:1147 physical and chemical stimuli 1119:1147 physical and chemical stimuli 1119:1147 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27991905 3 29 theme polycystin-specific 400:418 arg1 opening 432:438 a novel polycystin-specific 'tetragonal opening 392:438 a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure 392:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 0 30 theme channel 47:53 arg1 PC2 69:71 PC2 69:71 PC2 69:71 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 0 30 theme channel 47:53 arg1 Polycystin-2 55:66 TRP channel Polycystin-2 43:66 TRP channel Polycystin-2 (PC2) 43:72 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 5 31 theme missense 985:992 arg1 variants 994:1001 ADPKD-associated missense variants 968:1001 ADPKD-associated missense variants 968:1001 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 3 32 theme transient 508:516 arg1 channel 543:549 a classic transient receptor potential (TRP) channel 498:549 a classic transient receptor potential (TRP) channel structure 498:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 0 33 theme TRP 43:45 arg1 PC2 69:71 PC2 69:71 PC2 69:71 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 0 33 theme TRP 43:45 arg1 Polycystin-2 55:66 TRP channel Polycystin-2 43:66 TRP channel Polycystin-2 (PC2) 43:72 Structure of the polycystic kidney disease TRP channel Polycystin-2 (PC2). 27991905 3 34 theme receptor 518:525 arg1 channel 543:549 a classic transient receptor potential (TRP) channel 498:549 a classic transient receptor potential (TRP) channel structure 498:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 2 35 theme PC2 285:287 arg1 structure 266:274 the structure 262:274 the structure 262:274 Here we present the structure of human PC2 in a closed conformation, solved by electron cryomicroscopy at 4.2-Å resolution. 27991905 5 36 theme homologous 897:906 arg1 region 921:926 the homologous channel-like region 893:926 the homologous channel-like region of PC1 893:933 The TOP-domain fold is conserved among the polycystins, including the homologous channel-like region of PC1, and is the site of a cluster of ADPKD-associated missense variants. 27991905 3 37 theme channel 543:549 arg1 structure 551:559 a classic transient receptor potential (TRP) channel structure 498:559 a classic transient receptor potential (TRP) channel structure 498:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 2 38 theme human 279:283 arg1 PC2 285:287 human PC2 279:287 human PC2 279:287 Here we present the structure of human PC2 in a closed conformation, solved by electron cryomicroscopy at 4.2-Å resolution. 27991905 1 39 theme autosomal-dominant 165:182 arg1 ADPKD 211:215 ADPKD 211:215 ADPKD 211:215 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 39 theme autosomal-dominant 165:182 arg1 disease 202:208 autosomal-dominant polycystic kidney disease 165:208 autosomal-dominant polycystic kidney disease (ADPKD) 165:216 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 4 40 theme ion-conduction 803:816 arg1 pathway 818:824 the ion-conduction pathway 799:824 the ion-conduction pathway 799:824 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 3 41 theme structure 551:559 arg1 top 491:493 the top 487:493 the top of a classic transient receptor potential (TRP) channel structure 487:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 1 42 theme polycystic 184:193 arg1 ADPKD 211:215 ADPKD 211:215 ADPKD 211:215 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 42 theme polycystic 184:193 arg1 disease 202:208 autosomal-dominant polycystic kidney disease 165:208 autosomal-dominant polycystic kidney disease (ADPKD) 165:216 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 4 43 theme TOP 566:568 arg1 domain 570:575 The TOP domain 562:575 The TOP domain 562:575 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 3 44 theme TRP 538:540 arg1 channel 543:549 a classic transient receptor potential (TRP) channel 498:549 a classic transient receptor potential (TRP) channel structure 498:559 The structure reveals a novel polycystin-specific 'tetragonal opening for polycystins' (TOP) domain tightly bound to the top of a classic transient receptor potential (TRP) channel structure. 27991905 4 45 dep reticulum 685:693 arg1 lumen 695:699 lumen 695:699 lumen 695:699 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 1 46 from Mutations 75:83 arg1 PC1 109:111 PC1 or PKD1 109:119 PC1 109:111 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 46 from Mutations 75:83 arg1 polycystin-1 95:106 polycystin-1 95:106 polycystin-1 (PC1 or PKD1) 95:120 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 46 from Mutations 75:83 arg1 PKD1 116:119 PC1 or PKD1 109:119 PKD1 116:119 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 46 from Mutations 75:83 arg1 polycystin-2 125:136 polycystin-2 125:136 polycystin-2 (PC2, PKD2 or TRPP1) 125:157 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 47 theme kidney 195:200 arg1 ADPKD 211:215 ADPKD 211:215 ADPKD 211:215 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 1 47 theme kidney 195:200 arg1 disease 202:208 autosomal-dominant polycystic kidney disease 165:208 autosomal-dominant polycystic kidney disease (ADPKD) 165:216 Mutations in either polycystin-1 (PC1 or PKD1) or polycystin-2 (PC2, PKD2 or TRPP1) cause autosomal-dominant polycystic kidney disease (ADPKD) through unknown mechanisms. 27991905 4 48 theme extracellular 704:716 arg1 surface 718:724 extracellular surface 704:724 extracellular surface 704:724 The TOP domain is formed from two extensions to the voltage-sensor-like domain (VSLD); it covers the channel's endoplasmic reticulum lumen or extracellular surface and encloses an upper vestibule, above the pore filter, without blocking the ion-conduction pathway. 27991905 6 49 theme Extensive 1004:1012 arg1 contacts 1014:1021 Extensive contacts 1004:1021 Extensive contacts among the TOP-domain subunits, the pore and the VSLD 1004:1074 Extensive contacts among the TOP-domain subunits, the pore and the VSLD provide ample scope for regulation through physical and chemical stimuli. 27768895 2 0 theme lipid 292:296 arg1 bilayers 298:305 lipid bilayers 292:305 lipid bilayers 292:305 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 0 1 theme Lipid 63:67 arg1 Nanodiscs 69:77 Lipid Nanodiscs 63:77 Lipid Nanodiscs 63:77 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 0 2 from Structure 4:12 arg1 Nanodiscs 69:77 Lipid Nanodiscs 63:77 Lipid Nanodiscs 63:77 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 2 3 from structure 271:279 arg1 bilayers 298:305 lipid bilayers 292:305 lipid bilayers 292:305 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 6 4 theme molecular 1036:1044 arg1 basis 1046:1050 the molecular basis 1032:1050 the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD 1032:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 5 5 theme channel 822:828 arg1 assembly 830:837 channel assembly 822:837 channel assembly 822:837 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 3 6 located found 525:529 arg1 those 534:538 those 534:538 those 534:538 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 27768895 3 6 located found 525:529 arg2 charges 508:514 two of four gating charges 489:514 two of four gating charges commonly found in those of voltage-gated ion channels 489:568 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 27768895 1 7 theme common 208:213 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 8 theme human 215:219 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 2 9 theme PKD2 284:287 arg1 structure 271:279 the cryo-EM structure 259:279 the cryo-EM structure of PKD2 in lipid bilayers 259:305 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 4 10 theme ion 715:717 arg1 conduction 719:728 ion conduction 715:728 ion conduction 715:728 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 5 11 theme pathogenic 785:794 arg1 mutations 796:804 ADPKD pathogenic mutations 779:804 ADPKD pathogenic mutations 779:804 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 5 12 theme extracellular 735:747 arg1 PKD2 759:762 PKD2 759:762 PKD2 759:762 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 5 12 theme extracellular 735:747 arg1 domain 749:754 The extracellular domain 731:754 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, 731:805 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 2 13 theme potential 409:417 arg1 mechanisms 419:428 potential mechanisms 409:428 potential mechanisms 409:428 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 2 14 theme cryo-EM 263:269 arg1 structure 271:279 the cryo-EM structure 259:279 the cryo-EM structure of PKD2 in lipid bilayers 259:305 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 1 15 theme autosomal 138:146 arg1 one 192:194 one 192:194 one 192:194 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 15 theme autosomal 138:146 arg1 disease 175:181 autosomal dominant polycystic kidney disease 138:181 autosomal dominant polycystic kidney disease (ADPKD) 138:189 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 15 theme autosomal 138:146 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 15 theme autosomal 138:146 arg1 ADPKD 184:188 ADPKD 184:188 ADPKD 184:188 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 16 theme monogenic 221:229 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 5 17 theme transmembrane 876:888 arg1 core 890:893 the transmembrane core 872:893 the transmembrane core 872:893 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 1 18 theme dominant 148:155 arg1 one 192:194 one 192:194 one 192:194 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 18 theme dominant 148:155 arg1 disease 175:181 autosomal dominant polycystic kidney disease 138:181 autosomal dominant polycystic kidney disease (ADPKD) 138:189 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 18 theme dominant 148:155 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 18 theme dominant 148:155 arg1 ADPKD 184:188 ADPKD 184:188 ADPKD 184:188 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 2 19 attach present 251:257 arg2 we 248:249 we 248:249 we 248:249 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 2 19 attach present 251:257 arg1 resolution 316:325 3.0 Å resolution 310:325 3.0 Å resolution 310:325 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 4 20 theme S6 676:677 arg1 end 669:671 the cytoplasmic end 653:671 the cytoplasmic end of S6 653:677 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 1 21 theme polycystic 157:166 arg1 one 192:194 one 192:194 one 192:194 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 21 theme polycystic 157:166 arg1 disease 175:181 autosomal dominant polycystic kidney disease 138:181 autosomal dominant polycystic kidney disease (ADPKD) 138:189 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 21 theme polycystic 157:166 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 21 theme polycystic 157:166 arg1 ADPKD 184:188 ADPKD 184:188 ADPKD 184:188 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 2 22 theme ion 371:373 arg1 channel 375:381 a homotetrameric ion channel 354:381 a homotetrameric ion channel 354:381 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 2 22 theme ion 371:373 arg1 PKD2 346:349 PKD2 346:349 PKD2 346:349 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 1 23 theme disorders 231:239 arg1 one 192:194 one 192:194 one 192:194 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 23 theme disorders 231:239 arg1 disease 175:181 autosomal dominant polycystic kidney disease 138:181 autosomal dominant polycystic kidney disease (ADPKD) 138:189 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 23 theme disorders 231:239 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 5 24 theme extracellular 939:951 arg1 stimuli 953:959 extracellular stimuli 939:959 extracellular stimuli 939:959 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 1 25 theme Polycystic 84:93 arg1 gene 119:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 26 theme kidney 168:173 arg1 one 192:194 one 192:194 one 192:194 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 26 theme kidney 168:173 arg1 disease 175:181 autosomal dominant polycystic kidney disease 138:181 autosomal dominant polycystic kidney disease (ADPKD) 138:189 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 26 theme kidney 168:173 arg1 disorders 231:239 the most common human monogenic disorders 199:239 the most common human monogenic disorders 199:239 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 1 26 theme kidney 168:173 arg1 ADPKD 184:188 ADPKD 184:188 ADPKD 184:188 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 2 27 theme homotetrameric 356:369 arg1 channel 375:381 a homotetrameric ion channel 354:381 a homotetrameric ion channel 354:381 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 2 27 theme homotetrameric 356:369 arg1 PKD2 346:349 PKD2 346:349 PKD2 346:349 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 1 28 theme Kidney 95:100 arg1 gene 119:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 6 29 from ADPKD 1109:1113 arg1 majority 1060:1067 the majority 1056:1067 the majority of pathogenic mutations in Pkd2-related ADPKD 1056:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 0 30 theme Kidney 32:37 arg1 PKD2 55:58 the Polycystic Kidney Disease Channel PKD2 17:58 the Polycystic Kidney Disease Channel PKD2 17:58 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 2 31 theme 3.0 Å 310:314 arg1 resolution 316:325 3.0 Å resolution 310:325 3.0 Å resolution 310:325 Here, we present the cryo-EM structure of PKD2 in lipid bilayers at 3.0 Å resolution, which establishes PKD2 as a homotetrameric ion channel and provides insight into potential mechanisms for its activation. 27768895 4 32 theme permeation 584:593 arg1 pathway 595:601 The PKD2 ion permeation pathway 571:601 The PKD2 ion permeation pathway 571:601 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 0 33 theme Polycystic 21:30 arg1 PKD2 55:58 the Polycystic Kidney Disease Channel PKD2 17:58 the Polycystic Kidney Disease Channel PKD2 17:58 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 5 34 theme PKD2 759:762 arg1 PKD2 759:762 PKD2 759:762 PKD2 759:762 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 5 34 theme PKD2 759:762 arg1 domain 749:754 The extracellular domain 731:754 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, 731:805 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 4 35 theme ion 580:582 arg1 pathway 595:601 The PKD2 ion permeation pathway 571:601 The PKD2 ion permeation pathway 571:601 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 0 36 theme Channel 47:53 arg1 PKD2 55:58 the Polycystic Kidney Disease Channel PKD2 17:58 the Polycystic Kidney Disease Channel PKD2 17:58 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 6 37 from majority 1060:1067 arg1 ADPKD 1109:1113 Pkd2-related ADPKD 1096:1113 Pkd2-related ADPKD 1096:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 1 38 theme Disease 102:108 arg1 gene 119:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 0 39 theme Disease 39:45 arg1 PKD2 55:58 the Polycystic Kidney Disease Channel PKD2 17:58 the Polycystic Kidney Disease Channel PKD2 17:58 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 6 40 from mutations 1083:1091 arg1 ADPKD 1109:1113 Pkd2-related ADPKD 1096:1113 Pkd2-related ADPKD 1096:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 6 41 theme pathogenic 1072:1081 arg1 mutations 1083:1091 pathogenic mutations 1072:1091 pathogenic mutations in Pkd2-related ADPKD 1072:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 4 42 theme cytoplasmic 657:667 arg1 end 669:671 the cytoplasmic end 653:671 the cytoplasmic end of S6 653:677 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 4 43 theme the selectivity 621:635 arg1 filter 637:642 the selectivity filter 621:642 the selectivity filter 621:642 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 6 44 theme Pkd2-related 1096:1107 arg1 ADPKD 1109:1113 Pkd2-related ADPKD 1096:1113 Pkd2-related ADPKD 1096:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 5 45 theme physical 916:923 arg1 substrate 925:933 a physical substrate 914:933 a physical substrate for extracellular stimuli 914:959 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 1 46 theme Pkd2 113:116 arg1 gene 119:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene 80:122 The Polycystic Kidney Disease 2 (Pkd2) gene is mutated in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic disorders. 27768895 6 47 theme mutations 1083:1091 arg1 majority 1060:1067 the majority 1056:1067 the majority of pathogenic mutations in Pkd2-related ADPKD 1056:1113 Finally, our structure establishes the molecular basis for the majority of pathogenic mutations in Pkd2-related ADPKD. 27768895 3 48 theme gating 501:506 arg1 charges 508:514 two of four gating charges 489:514 two of four gating charges commonly found in those of voltage-gated ion channels 489:568 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 27768895 5 49 theme ADPKD 779:783 arg1 mutations 796:804 ADPKD pathogenic mutations 779:804 ADPKD pathogenic mutations 779:804 The extracellular domain of PKD2, a hotspot for ADPKD pathogenic mutations, contributes to channel assembly and strategically interacts with the transmembrane core, likely serving as a physical substrate for extracellular stimuli to allosterically gate the channel. 27768895 3 50 theme voltage-gated 543:555 arg1 channels 561:568 voltage-gated ion channels 543:568 voltage-gated ion channels 543:568 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 27768895 4 51 theme PKD2 575:578 arg1 pathway 595:601 The PKD2 ion permeation pathway 571:601 The PKD2 ion permeation pathway 571:601 The PKD2 ion permeation pathway is constricted at the selectivity filter and near the cytoplasmic end of S6, suggesting that two gates regulate ion conduction. 27768895 3 52 theme PKD2 454:457 arg1 domain 474:479 The PKD2 voltage-sensor domain 450:479 The PKD2 voltage-sensor domain 450:479 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 27768895 0 53 theme PKD2 55:58 arg1 Structure 4:12 The Structure 0:12 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 0:78 The Structure of the Polycystic Kidney Disease Channel PKD2 in Lipid Nanodiscs. 27768895 3 54 theme ion 557:559 arg1 channels 561:568 voltage-gated ion channels 543:568 voltage-gated ion channels 543:568 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 27768895 3 55 theme voltage-sensor 459:472 arg1 domain 474:479 The PKD2 voltage-sensor domain 450:479 The PKD2 voltage-sensor domain 450:479 The PKD2 voltage-sensor domain retains two of four gating charges commonly found in those of voltage-gated ion channels. 8939990 9 0 theme family 1576:1581 arg1 IGFBP-7 1584:1590 IGFBP-7 1584:1590 IGFBP-7 1584:1590 In conclusion, mac25 specifically binds IGFs and constitutes a new member of the IGFBP family, IGFBP-7. 8939990 9 0 theme family 1576:1581 arg1 member 1556:1561 a new member 1550:1561 a new member of the IGFBP family 1550:1581 In conclusion, mac25 specifically binds IGFs and constitutes a new member of the IGFBP family, IGFBP-7. 8939990 10 1 from distribution 1603:1614 arg1 cells 1672:1676 several cancer cells 1657:1676 several cancer cells 1657:1676 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 10 1 from distribution 1603:1614 arg1 tissue 1626:1631 normal tissue 1619:1631 normal tissue 1619:1631 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 5 2 theme ligand 879:884 arg1 blotting 886:893 Western ligand blotting 871:893 Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis 871:948 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 4 3 theme present 582:588 arg1 study 590:594 the present study 578:594 the present study 578:594 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 6 4 theme rh-mac25 1094:1101 arg1 binding 1103:1109 rh-mac25 binding 1094:1109 rh-mac25 binding 1094:1109 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 10 5 from expression 1643:1652 arg1 cells 1672:1676 several cancer cells 1657:1676 several cancer cells 1657:1676 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 10 5 from expression 1643:1652 arg1 tissue 1626:1631 normal tissue 1619:1631 normal tissue 1619:1631 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 4 6 from expression 754:763 arg1 tissues 792:798 human tissues 786:798 human tissues 786:798 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 1 7 theme mac25 106:110 arg1 protein 112:118 Recombinant human mac25 protein 88:118 Recombinant human mac25 protein 88:118 Recombinant human mac25 protein specifically binds IGF-I and -II. 8939990 6 8 theme IGF-II 1152:1157 arg1 analog 1159:1164 [QAYLL]IGF-II analog 1145:1164 [QAYLL]IGF-II analog 1145:1164 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 8 9 theme cancer 1470:1475 arg1 lines 1482:1486 cancer cell lines 1470:1486 cancer cell lines 1470:1486 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 8 10 from range 1376:1380 arg1 detectable 1355:1364 detectable 1355:1364 detectable 1355:1364 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 5 11 theme gel 930:932 arg1 electrophoresis 934:948 polyacrylamide gel electrophoresis 915:948 polyacrylamide gel electrophoresis 915:948 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 3 12 theme amino 370:374 arg1 sequence 381:388 The deduced amino acid sequence 358:388 The deduced amino acid sequence of the human mac25 propeptide 358:418 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 7 13 contain has 1249:1251 arg2 affinity 1319:1326 20-25-fold lower affinity 1302:1326 20-25-fold lower affinity for IGF-II 1302:1337 In comparison with IGFBP-3, rh-mac25 has at least a 5-6-fold lower affinity for IGF-I and 20-25-fold lower affinity for IGF-II. 8939990 7 13 contain has 1249:1251 arg1 rh-mac25 1240:1247 rh-mac25 1240:1247 rh-mac25 1240:1247 In comparison with IGFBP-3, rh-mac25 has at least a 5-6-fold lower affinity for IGF-I and 20-25-fold lower affinity for IGF-II. 8939990 7 13 contain has 1249:1251 arg2 affinity 1279:1286 a 5-6-fold lower affinity 1262:1286 a 5-6-fold lower affinity for IGF-I 1262:1296 In comparison with IGFBP-3, rh-mac25 has at least a 5-6-fold lower affinity for IGF-I and 20-25-fold lower affinity for IGF-II. 8939990 5 14 theme 125I-IGF-I 812:821 arg1 Binding 801:807 Binding 801:807 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 801:849 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 4 15 theme human 627:631 arg1 rh-mac25 640:647 rh-mac25 640:647 rh-mac25 640:647 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 4 15 theme human 627:631 arg1 mac25 633:637 recombinant human mac25 615:637 recombinant human mac25 (rh-mac25) 615:648 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 8 16 with detectable 1355:1364 arg1 expression 1422:1431 decreased expression 1412:1431 decreased expression in breast, prostate, colon, and lung cancer cell lines 1412:1486 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 3 17 theme human 448:452 arg1 IGFBPs 499:504 IGFBPs 499:504 IGFBPs 499:504 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 3 17 theme human 448:452 arg1 proteins 489:496 human insulin-like growth factor-binding proteins 448:496 human insulin-like growth factor-binding proteins (IGFBPs) 448:505 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 3 18 theme acid 376:379 arg1 sequence 381:388 The deduced amino acid sequence 358:388 The deduced amino acid sequence of the human mac25 propeptide 358:418 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 0 19 theme growth 47:52 arg1 IGFBP 78:82 insulin-like growth factor-binding protein (IGFBP)-7 34:85 insulin-like growth factor-binding protein (IGFBP)-7 34:85 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. 8939990 6 20 theme [QAYLL 1145:1150 arg1 analog 1159:1164 [QAYLL]IGF-II analog 1145:1164 [QAYLL]IGF-II analog 1145:1164 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 4 21 theme expression 754:763 arg1 pattern 743:749 the pattern 739:749 the pattern of expression of the mac25 gene in human tissues 739:798 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 3 22 theme insulin-like 454:465 arg1 IGFBPs 499:504 IGFBPs 499:504 IGFBPs 499:504 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 3 22 theme insulin-like 454:465 arg1 proteins 489:496 human insulin-like growth factor-binding proteins 448:496 human insulin-like growth factor-binding proteins (IGFBPs) 448:505 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 3 23 theme mac25 403:407 arg1 propeptide 409:418 the human mac25 propeptide 393:418 the human mac25 propeptide 393:418 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 8 24 theme cell 1477:1480 arg1 lines 1482:1486 cancer cell lines 1470:1486 cancer cell lines 1470:1486 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 8 25 from expression 1422:1431 arg1 breast 1436:1441 breast 1436:1441 breast 1436:1441 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 8 25 from expression 1422:1431 arg1 colon 1454:1458 colon 1454:1458 colon 1454:1458 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 8 25 from expression 1422:1431 arg1 lung 1465:1468 lung 1465:1468 lung 1465:1468 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 8 25 from expression 1422:1431 arg1 prostate 1444:1451 prostate 1444:1451 prostate 1444:1451 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 2 26 theme epithelial 340:349 arg1 cells 351:355 senescent human mammary epithelial cells 316:355 senescent human mammary epithelial cells 316:355 The mac25 cDNA was originally cloned from leptomeningial cells and subsequently reisolated through differential display as a sequence preferentially expressed in senescent human mammary epithelial cells. 8939990 9 27 theme new 1552:1554 arg1 IGFBP-7 1584:1590 IGFBP-7 1584:1590 IGFBP-7 1584:1590 In conclusion, mac25 specifically binds IGFs and constitutes a new member of the IGFBP family, IGFBP-7. 8939990 9 27 theme new 1552:1554 arg1 member 1556:1561 a new member 1550:1561 a new member of the IGFBP family 1550:1581 In conclusion, mac25 specifically binds IGFs and constitutes a new member of the IGFBP family, IGFBP-7. 8939990 4 28 theme baculovirus 655:665 arg1 system 678:683 a baculovirus expression system 653:683 a baculovirus expression system 653:683 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 3 29 theme propeptide 409:418 arg1 sequence 381:388 The deduced amino acid sequence 358:388 The deduced amino acid sequence of the human mac25 propeptide 358:418 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 3 30 theme family 567:572 arg1 member 547:552 another member 539:552 another member of the IGFBP family 539:572 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 10 31 theme normal 1619:1624 arg1 tissue 1626:1631 normal tissue 1619:1631 normal tissue 1619:1631 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 1 32 theme Recombinant 88:98 arg1 protein 112:118 Recombinant human mac25 protein 88:118 Recombinant human mac25 protein 88:118 Recombinant human mac25 protein specifically binds IGF-I and -II. 8939990 7 33 with comparison 1215:1224 arg1 IGFBP-3 1231:1237 IGFBP-3 1231:1237 IGFBP-3 1231:1237 In comparison with IGFBP-3, rh-mac25 has at least a 5-6-fold lower affinity for IGF-I and 20-25-fold lower affinity for IGF-II. 8939990 5 34 with cross-linking 966:978 arg1 nM 1000:1001 as little as 2 nM rh-mac25 985:1010 as little as 2 nM rh-mac25 985:1010 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 8 35 theme tissues 1398:1404 arg1 range 1376:1380 a wide range 1369:1380 a wide range of normal human tissues 1369:1404 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 8 36 theme decreased 1412:1420 arg1 expression 1422:1431 decreased expression 1412:1431 decreased expression in breast, prostate, colon, and lung cancer cell lines 1412:1486 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 4 37 from pattern 743:749 arg1 tissues 792:798 human tissues 786:798 human tissues 786:798 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 2 38 theme human 326:330 arg1 cells 351:355 senescent human mammary epithelial cells 316:355 senescent human mammary epithelial cells 316:355 The mac25 cDNA was originally cloned from leptomeningial cells and subsequently reisolated through differential display as a sequence preferentially expressed in senescent human mammary epithelial cells. 8939990 2 39 theme mammary 332:338 arg1 cells 351:355 senescent human mammary epithelial cells 316:355 senescent human mammary epithelial cells 316:355 The mac25 cDNA was originally cloned from leptomeningial cells and subsequently reisolated through differential display as a sequence preferentially expressed in senescent human mammary epithelial cells. 8939990 8 40 dep breast 1436:1441 arg1 lines 1482:1486 cancer cell lines 1470:1486 cancer cell lines 1470:1486 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 10 41 theme cancer 1665:1670 arg1 cells 1672:1676 several cancer cells 1657:1676 several cancer cells 1657:1676 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 2 42 theme senescent 316:324 arg1 cells 351:355 senescent human mammary epithelial cells 316:355 senescent human mammary epithelial cells 316:355 The mac25 cDNA was originally cloned from leptomeningial cells and subsequently reisolated through differential display as a sequence preferentially expressed in senescent human mammary epithelial cells. 8939990 3 43 theme human 397:401 arg1 propeptide 409:418 the human mac25 propeptide 393:418 the human mac25 propeptide 393:418 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 3 44 theme 20-25 429:433 arg1 % 434:434 % 434:434 % 434:434 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 7 45 theme lower 1273:1277 arg1 affinity 1279:1286 a 5-6-fold lower affinity 1262:1286 a 5-6-fold lower affinity for IGF-I 1262:1296 In comparison with IGFBP-3, rh-mac25 has at least a 5-6-fold lower affinity for IGF-I and 20-25-fold lower affinity for IGF-II. 8939990 10 46 theme growth-suppressing 1718:1735 arg1 IGFBP-7 1692:1698 IGFBP-7 1692:1698 IGFBP-7 1692:1698 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 10 46 theme growth-suppressing 1718:1735 arg1 factor 1737:1742 a growth-suppressing factor 1716:1742 a growth-suppressing factor 1716:1742 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 1 47 theme human 100:104 arg1 protein 112:118 Recombinant human mac25 protein 88:118 Recombinant human mac25 protein 88:118 Recombinant human mac25 protein specifically binds IGF-I and -II. 8939990 10 48 theme wider 1597:1601 arg1 distribution 1603:1614 Its wider distribution 1593:1614 Its wider distribution in normal tissue 1593:1631 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 0 49 theme insulin-like 34:45 arg1 IGFBP 78:82 insulin-like growth factor-binding protein (IGFBP)-7 34:85 insulin-like growth factor-binding protein (IGFBP)-7 34:85 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. 8939990 3 50 theme % 434:434 arg1 identity 436:443 a 20-25% identity 427:443 a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs) 427:505 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 4 51 theme human 786:790 arg1 tissues 792:798 human tissues 786:798 human tissues 786:798 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 5 52 theme 125I-IGF-II 827:837 arg1 Binding 801:807 Binding 801:807 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 801:849 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 0 53 theme protein 69:75 arg1 IGFBP 78:82 insulin-like growth factor-binding protein (IGFBP)-7 34:85 insulin-like growth factor-binding protein (IGFBP)-7 34:85 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. 8939990 10 54 theme several 1657:1663 arg1 cells 1672:1676 several cancer cells 1657:1676 several cancer cells 1657:1676 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 6 55 with competition 1078:1088 arg1 analog 1159:1164 [QAYLL]IGF-II analog 1145:1164 [QAYLL]IGF-II analog 1145:1164 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 6 55 with competition 1078:1088 arg1 IGFs 1126:1129 unlabeled IGFs 1116:1129 unlabeled IGFs 1116:1129 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 6 56 theme unlabeled 1116:1124 arg1 IGFs 1126:1129 unlabeled IGFs 1116:1129 unlabeled IGFs 1116:1129 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 10 57 theme lower 1637:1641 arg1 expression 1643:1652 lower expression 1637:1652 lower expression in several cancer cells 1637:1676 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 0 58 theme factor-binding 54:67 arg1 IGFBP 78:82 insulin-like growth factor-binding protein (IGFBP)-7 34:85 insulin-like growth factor-binding protein (IGFBP)-7 34:85 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. 8939990 10 59 theme IGF-binding 1759:1769 arg1 protein 1771:1777 an IGF-binding protein 1756:1777 an IGF-binding protein 1756:1777 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 10 59 theme IGF-binding 1759:1769 arg1 IGFBP-7 1692:1698 IGFBP-7 1692:1698 IGFBP-7 1692:1698 Its wider distribution in normal tissue and lower expression in several cancer cells indicate that IGFBP-7 may function as a growth-suppressing factor, as well as an IGF-binding protein. 8939990 3 60 dep insulin-like 454:465 arg1 factor-binding 474:487 factor-binding 474:487 factor-binding 474:487 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 4 61 from tissues 792:798 arg1 pattern 743:749 the pattern 739:749 the pattern of expression of the mac25 gene in human tissues 739:798 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 8 62 theme normal 1385:1390 arg1 tissues 1398:1404 normal human tissues 1385:1404 normal human tissues 1385:1404 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 4 63 theme gene 778:781 arg1 expression 754:763 expression 754:763 expression of the mac25 gene in human tissues 754:798 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 2 64 theme leptomeningial 196:209 arg1 cells 211:215 leptomeningial cells 196:215 leptomeningial cells 196:215 The mac25 cDNA was originally cloned from leptomeningial cells and subsequently reisolated through differential display as a sequence preferentially expressed in senescent human mammary epithelial cells. 8939990 4 65 theme mac25 772:776 arg1 gene 778:781 the mac25 gene 768:781 the mac25 gene 768:781 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 9 66 theme IGFBP 1570:1574 arg1 family 1576:1581 the IGFBP family 1566:1581 the IGFBP family 1566:1581 In conclusion, mac25 specifically binds IGFs and constitutes a new member of the IGFBP family, IGFBP-7. 8939990 5 67 theme polyacrylamide 915:928 arg1 electrophoresis 934:948 polyacrylamide gel electrophoresis 915:948 polyacrylamide gel electrophoresis 915:948 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 3 68 theme IGFBP 561:565 arg1 family 567:572 the IGFBP family 557:572 the IGFBP family 557:572 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 6 69 theme rh-mac25 1028:1035 arg1 binding 1037:1043 rh-mac25 binding 1028:1043 rh-mac25 binding to 125I-IGFs 1028:1056 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 8 70 theme human 1392:1396 arg1 tissues 1398:1404 normal human tissues 1385:1404 normal human tissues 1385:1404 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 5 71 theme Western 871:877 arg1 blotting 886:893 Western ligand blotting 871:893 Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis 871:948 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 8 72 theme mac25 1340:1344 arg1 mRNA 1346:1349 mac25 mRNA 1340:1349 mac25 mRNA 1340:1349 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 0 73 theme IGFBP 78:82 arg1 characterization 14:29 characterization 14:29 characterization 14:29 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. 8939990 0 73 theme IGFBP 78:82 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis 0:8 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. 8939990 6 74 theme binding 1037:1043 arg1 Specificity 1013:1023 Specificity 1013:1023 Specificity of rh-mac25 binding to 125I-IGFs 1013:1056 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 6 75 theme less 1186:1189 arg1 affinity 1191:1198 100-fold less affinity 1177:1198 100-fold less affinity for IGFBPs 1177:1209 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 8 76 from detectable 1355:1364 arg1 range 1376:1380 a wide range 1369:1380 a wide range of normal human tissues 1369:1404 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 5 77 theme affinity 957:964 arg1 cross-linking 966:978 affinity cross-linking 957:978 affinity cross-linking with as little as 2 nM rh-mac25 957:1010 Binding of 125I-IGF-I and 125I-IGF-II to rh-mac25 was demonstrated by Western ligand blotting after nondenaturing polyacrylamide gel electrophoresis and by affinity cross-linking with as little as 2 nM rh-mac25. 8939990 7 78 theme lower 1313:1317 arg1 affinity 1319:1326 20-25-fold lower affinity 1302:1326 20-25-fold lower affinity for IGF-II 1302:1337 In comparison with IGFBP-3, rh-mac25 has at least a 5-6-fold lower affinity for IGF-I and 20-25-fold lower affinity for IGF-II. 8939990 4 79 theme recombinant 615:625 arg1 rh-mac25 640:647 rh-mac25 640:647 rh-mac25 640:647 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 4 79 theme recombinant 615:625 arg1 mac25 633:637 recombinant human mac25 615:637 recombinant human mac25 (rh-mac25) 615:648 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 3 80 theme deduced 362:368 arg1 sequence 381:388 The deduced amino acid sequence 358:388 The deduced amino acid sequence of the human mac25 propeptide 358:418 The deduced amino acid sequence of the human mac25 propeptide shares a 20-25% identity to human insulin-like growth factor-binding proteins (IGFBPs), suggesting that mac25 could be another member of the IGFBP family. 8939990 2 81 theme differential 253:264 arg1 display 266:272 differential display 253:272 differential display as a sequence preferentially expressed in senescent human mammary epithelial cells 253:355 The mac25 cDNA was originally cloned from leptomeningial cells and subsequently reisolated through differential display as a sequence preferentially expressed in senescent human mammary epithelial cells. 8939990 4 82 theme expression 667:676 arg1 system 678:683 a baculovirus expression system 653:683 a baculovirus expression system 653:683 In the present study, we have generated recombinant human mac25 (rh-mac25) in a baculovirus expression system and assessed its affinity for IGFs and have evaluated the pattern of expression of the mac25 gene in human tissues. 8939990 8 83 theme wide 1371:1374 arg1 range 1376:1380 a wide range 1369:1380 a wide range of normal human tissues 1369:1404 mac25 mRNA was detectable in a wide range of normal human tissues, with decreased expression in breast, prostate, colon, and lung cancer cell lines. 8939990 6 84 contain has 1173:1175 arg1 analog 1159:1164 [QAYLL]IGF-II analog 1145:1164 [QAYLL]IGF-II analog 1145:1164 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 8939990 6 84 contain has 1173:1175 arg2 affinity 1191:1198 100-fold less affinity 1177:1198 100-fold less affinity for IGFBPs 1177:1209 Specificity of rh-mac25 binding to 125I-IGFs was demonstrated by competition for rh-mac25 binding with unlabeled IGFs, but not with [QAYLL]IGF-II analog, which has 100-fold less affinity for IGFBPs. 18768470 2 0 theme growth 150:155 arg1 beta 164:167 Transforming growth factor beta 137:167 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 6 1 theme A 1269:1269 arg1 affinity 1271:1278 activin A affinity 1261:1278 activin A affinity 1261:1278 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 8 2 theme N-terminal 1523:1532 arg1 domain 1534:1539 the N-terminal domain 1519:1539 the N-terminal domain 1519:1539 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 4 3 theme x-ray 603:607 arg1 structure 609:617 the x-ray structure 599:617 the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites 599:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 2 4 theme Transforming 137:148 arg1 beta 164:167 Transforming growth factor beta 137:167 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 6 5 theme activin 1261:1267 arg1 A 1269:1269 activin A 1261:1269 activin A affinity 1261:1278 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 5 6 theme ligand 910:915 arg1 site 933:936 the ligand type I receptor site 906:936 the ligand type I receptor site 906:936 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 7 7 with interaction 1355:1365 arg1 A 1414:1414 activin A 1406:1414 activin A 1406:1414 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 7 8 theme significant 1343:1353 arg1 interaction 1355:1365 a significant interaction 1341:1365 a significant interaction of the N-terminal domain of FSTL3 with activin A 1341:1414 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 6 9 theme N-terminal 1190:1199 arg1 domain 1201:1206 the FSTL3 N-terminal domain 1180:1206 the FSTL3 N-terminal domain 1180:1206 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 8 10 theme key 1558:1560 arg1 site 1562:1565 the key site 1554:1565 the key site for determining FS-type antagonist specificity 1554:1612 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 8 10 theme key 1558:1560 arg1 interface 1503:1511 the interface 1499:1511 the interface where the N-terminal domain binds 1499:1545 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 8 11 theme bone 1458:1461 arg1 proteins 1477:1484 bone morphogenetic proteins 1458:1484 bone morphogenetic proteins 1458:1484 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 3 12 theme high 381:384 arg1 affinity 386:393 high affinity 381:393 high affinity for activin A 381:407 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 13 theme Follistatin-type 251:266 arg1 antagonists 268:278 Follistatin-type antagonists 251:278 Follistatin-type antagonists 251:278 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 6 14 theme binding 1140:1146 arg1 studies 1148:1154 binding studies 1140:1154 binding studies 1140:1154 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 5 15 theme type 917:920 arg1 site 933:936 the ligand type I receptor site 906:936 the ligand type I receptor site 906:936 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 3 16 theme splice 295:300 arg1 FS315 337:341 FS315 337:341 FS315 337:341 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 16 theme splice 295:300 arg1 FS288 327:331 FS288 327:331 FS288 327:331 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 16 theme splice 295:300 arg1 variants 302:309 splice variants 295:309 splice variants of follistatin (FS288 and FS315) 295:342 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 1 17 theme N-terminal 66:75 arg1 domains 77:83 N-terminal domains 66:83 N-terminal domains 66:83 Differential binding of N-terminal domains influences follistatin-type antagonist specificity. 18768470 5 18 theme N-terminal 873:882 arg1 interaction 891:901 the FS N-terminal domain interaction 866:901 the FS N-terminal domain interaction 866:901 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 3 19 theme other 442:446 arg1 ligands 448:454 other ligands 442:454 other ligands 442:454 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 2 20 theme family 169:174 arg1 ligands 176:182 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 6 21 theme FS 1231:1232 arg1 domain 1234:1239 the corresponding FS domain 1213:1239 the corresponding FS domain 1213:1239 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 8 22 theme FS-type 1583:1589 arg1 specificity 1602:1612 FS-type antagonist specificity 1583:1612 FS-type antagonist specificity 1583:1612 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 1 23 theme domains 77:83 arg1 binding 55:61 Differential binding 42:61 Differential binding of N-terminal domains 42:83 Differential binding of N-terminal domains influences follistatin-type antagonist specificity. 18768470 5 24 theme FS 870:871 arg1 interaction 891:901 the FS N-terminal domain interaction 866:901 the FS N-terminal domain interaction 866:901 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 3 25 theme activin 399:405 arg1 A 407:407 activin A 399:407 activin A 399:407 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 2 26 theme beta 164:167 arg1 ligands 176:182 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 6 27 theme corresponding 1217:1229 arg1 domain 1234:1239 the corresponding FS domain 1213:1239 the corresponding FS domain 1213:1239 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 5 28 from site 933:936 arg1 significance 850:861 the significance 846:861 the significance of the FS N-terminal domain interaction at the ligand type I receptor site 846:936 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 7 29 theme FSTL3 1395:1399 arg1 FSTL3 1395:1399 FSTL3 1395:1399 FSTL3 1395:1399 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 7 29 theme FSTL3 1395:1399 arg1 domain 1385:1390 the N-terminal domain 1370:1390 the N-terminal domain of FSTL3 1370:1399 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 4 30 theme A. 677:678 arg1 resolution 659:668 a resolution 657:668 a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites 657:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 8 31 theme morphogenetic 1463:1475 arg1 proteins 1477:1484 bone morphogenetic proteins 1458:1484 bone morphogenetic proteins 1458:1484 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 1 32 theme follistatin-type 96:111 arg1 specificity 124:134 follistatin-type antagonist specificity 96:134 follistatin-type antagonist specificity 96:134 Differential binding of N-terminal domains influences follistatin-type antagonist specificity. 18768470 5 33 theme domain 884:889 arg1 interaction 891:901 the FS N-terminal domain interaction 866:901 the FS N-terminal domain interaction 866:901 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 0 34 theme FSTL3.activin 17:29 arg1 complex 33:39 FSTL3.activin A complex 17:39 FSTL3.activin A complex 17:39 The structure of FSTL3.activin A complex. 18768470 8 35 theme structural 1430:1439 arg1 comparisons 1441:1451 structural comparisons 1430:1451 structural comparisons with bone morphogenetic proteins 1430:1484 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 1 36 theme antagonist 113:122 arg1 specificity 124:134 follistatin-type antagonist specificity 96:134 follistatin-type antagonist specificity 96:134 Differential binding of N-terminal domains influences follistatin-type antagonist specificity. 18768470 3 37 theme bone 470:473 arg1 proteins 489:496 bone morphogenetic proteins 470:496 particularly bone morphogenetic proteins 457:496 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 37 theme bone 470:473 arg1 affinity 429:436 their affinity 423:436 their affinity for other ligands 423:454 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 4 38 theme antagonist 771:780 arg1 molecules 782:790 two antagonist molecules 767:790 two antagonist molecules blocking all ligand receptor-binding sites 767:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 5 39 with contact 1042:1048 arg1 A 1063:1063 activin A 1055:1063 activin A 1055:1063 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 3 40 theme morphogenetic 475:487 arg1 proteins 489:496 bone morphogenetic proteins 470:496 particularly bone morphogenetic proteins 457:496 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 40 theme morphogenetic 475:487 arg1 affinity 429:436 their affinity 423:436 their affinity for other ligands 423:454 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 0 41 theme complex 33:39 arg1 structure 4:12 The structure 0:12 The structure of FSTL3.activin A complex 0:39 The structure of FSTL3.activin A complex. 18768470 4 42 with complex 635:641 arg1 FSTL3 648:652 FSTL3 648:652 FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites 648:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 7 43 theme structural 1297:1306 arg1 evidence 1324:1331 both structural and biochemical evidence 1292:1331 both structural and biochemical evidence 1292:1331 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 0 44 theme A 31:31 arg1 complex 33:39 FSTL3.activin A complex 17:39 FSTL3.activin A complex 17:39 The structure of FSTL3.activin A complex. 18768470 3 45 theme follistatin 314:324 arg1 FS315 337:341 FS315 337:341 FS315 337:341 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 45 theme follistatin 314:324 arg1 FS288 327:331 FS288 327:331 FS288 327:331 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 45 theme follistatin 314:324 arg1 variants 302:309 splice variants 295:309 splice variants of follistatin (FS288 and FS315) 295:342 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 5 46 theme activin 1055:1061 arg1 A 1063:1063 activin A 1055:1063 activin A 1055:1063 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 4 47 theme receptor-binding 812:827 arg1 sites 829:833 all ligand receptor-binding sites 801:833 all ligand receptor-binding sites 801:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 7 48 theme activin 1406:1412 arg1 A 1414:1414 activin A 1406:1414 activin A 1406:1414 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 4 49 theme ligand 538:543 arg1 specificity 545:555 ligand specificity 538:555 ligand specificity within FS-type antagonists 538:582 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 7 50 theme N-terminal 1374:1383 arg1 FSTL3 1395:1399 FSTL3 1395:1399 FSTL3 1395:1399 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 7 50 theme N-terminal 1374:1383 arg1 domain 1385:1390 the N-terminal domain 1370:1390 the N-terminal domain of FSTL3 1370:1399 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 5 51 theme N-terminal 1002:1011 arg1 domain 1013:1018 the N-terminal domain 998:1018 the N-terminal domain 998:1018 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 4 52 theme FS.activin 715:724 arg1 structures 728:737 the previously resolved FS.activin A structures 691:737 the previously resolved FS.activin A structures 691:737 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 3 53 dep variants 302:309 arg1 FS315 337:341 FS315 337:341 FS315 337:341 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 53 dep variants 302:309 arg1 FS288 327:331 FS288 327:331 FS288 327:331 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 53 dep variants 302:309 arg1 variants 302:309 splice variants 295:309 splice variants of follistatin (FS288 and FS315) 295:342 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 4 54 theme FS-type 564:570 arg1 antagonists 572:582 FS-type antagonists 564:582 FS-type antagonists 564:582 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 4 55 theme structural 517:526 arg1 basis 528:532 the structural basis 513:532 the structural basis for ligand specificity within FS-type antagonists 513:582 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 2 56 theme antagonists 238:248 arg1 number 205:210 a number 203:210 a number of structurally divergent antagonists 203:248 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 6 57 theme FSTL3 1184:1188 arg1 domain 1201:1206 the FSTL3 N-terminal domain 1180:1206 the FSTL3 N-terminal domain 1180:1206 Furthermore, binding studies revealed that replacing the FSTL3 N-terminal domain with the corresponding FS domain considerably lowers activin A affinity. 18768470 4 58 theme resolved 706:713 arg1 structures 728:737 the previously resolved FS.activin A structures 691:737 the previously resolved FS.activin A structures 691:737 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 2 59 theme divergent 228:236 arg1 antagonists 238:248 structurally divergent antagonists 215:248 structurally divergent antagonists 215:248 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 4 60 dep A. 677:678 arg1 encircled 754:762 encircled 754:762 is encircled by two antagonist molecules blocking all ligand receptor-binding sites 751:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 3 61 contain have 376:379 arg1 antagonists 268:278 Follistatin-type antagonists 251:278 Follistatin-type antagonists 251:278 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 3 61 contain have 376:379 arg2 affinity 386:393 high affinity 381:393 high affinity for activin A 381:407 Follistatin-type antagonists, which include splice variants of follistatin (FS288 and FS315) and follistatin-like 3 (FSTL3), have high affinity for activin A but differ in their affinity for other ligands, particularly bone morphogenetic proteins. 18768470 5 62 theme receptor 924:931 arg1 site 933:936 the ligand type I receptor site 906:936 the ligand type I receptor site 906:936 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 7 63 theme domain 1385:1390 arg1 interaction 1355:1365 a significant interaction 1341:1365 a significant interaction of the N-terminal domain of FSTL3 with activin A 1341:1414 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 4 64 theme A 630:630 arg1 structure 609:617 the x-ray structure 599:617 the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites 599:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 5 65 dep questioned 947:956 arg1 show 977:980 show 977:980 show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS 977:1124 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 4 66 theme A 726:726 arg1 structures 728:737 the previously resolved FS.activin A structures 691:737 the previously resolved FS.activin A structures 691:737 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 4 67 theme activin 622:628 arg1 A 630:630 activin A 622:630 activin A 622:630 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 8 68 with comparisons 1441:1451 arg1 proteins 1477:1484 bone morphogenetic proteins 1458:1484 bone morphogenetic proteins 1458:1484 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 5 69 theme intimate 1033:1040 arg1 contact 1042:1048 a more intimate contact 1026:1048 a more intimate contact with activin A 1026:1063 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 4 70 theme ligand 805:810 arg1 sites 829:833 all ligand receptor-binding sites 801:833 all ligand receptor-binding sites 801:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 18768470 5 71 theme interaction 891:901 arg1 significance 850:861 the significance 846:861 the significance of the FS N-terminal domain interaction at the ligand type I receptor site 846:936 Recently, the significance of the FS N-terminal domain interaction at the ligand type I receptor site has been questioned; however, our data show that for FSTL3, the N-terminal domain forms a more intimate contact with activin A, implying that this interaction is stronger than that for FS. 18768470 7 72 theme biochemical 1312:1322 arg1 evidence 1324:1331 both structural and biochemical evidence 1292:1331 both structural and biochemical evidence 1292:1331 Therefore, both structural and biochemical evidence support a significant interaction of the N-terminal domain of FSTL3 with activin A. 18768470 2 73 theme factor 157:162 arg1 beta 164:167 Transforming growth factor beta 137:167 Transforming growth factor beta family ligands 137:182 Transforming growth factor beta family ligands are neutralized by a number of structurally divergent antagonists. 18768470 1 74 theme Differential 42:53 arg1 binding 55:61 Differential binding 42:61 Differential binding of N-terminal domains 42:83 Differential binding of N-terminal domains influences follistatin-type antagonist specificity. 18768470 8 75 theme antagonist 1591:1600 arg1 specificity 1602:1612 FS-type antagonist specificity 1583:1612 FS-type antagonist specificity 1583:1612 In addition, structural comparisons with bone morphogenetic proteins suggest that the interface where the N-terminal domain binds may be the key site for determining FS-type antagonist specificity. 18768470 4 76 from structure 609:617 arg1 complex 635:641 complex 635:641 complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites 635:833 To understand the structural basis for ligand specificity within FS-type antagonists, we determined the x-ray structure of activin A in complex with FSTL3 to a resolution of 2.5 A. Similar to the previously resolved FS.activin A structures, the ligand is encircled by two antagonist molecules blocking all ligand receptor-binding sites. 7477400 4 0 theme peptides 674:681 arg1 loading 653:659 the loading 649:659 the loading of antigenic peptides by an exchange process catalysed by DM 649:720 The structure is the substrate for the loading of antigenic peptides by an exchange process catalysed by DM. 7477400 0 1 from structure 4:12 arg1 maturation 49:58 class II MHC maturation 36:58 class II MHC maturation 36:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 4 2 theme antigenic 664:672 arg1 peptides 674:681 antigenic peptides 664:681 antigenic peptides 664:681 The structure is the substrate for the loading of antigenic peptides by an exchange process catalysed by DM. 7477400 1 3 theme antigen 205:211 arg1 presentation 213:224 antigen presentation 205:224 antigen presentation 205:224 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 1 4 theme invariant 128:136 arg1 chain 138:142 invariant chain 128:142 invariant chain called CLIP 128:154 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 2 5 theme histocompatibility 342:359 arg1 maturation 361:370 class II histocompatibility maturation 333:370 class II histocompatibility maturation 333:370 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 2 6 theme peptide-DR3 425:435 arg1 complexes 437:445 antigenic peptide-DR3 complexes 415:445 antigenic peptide-DR3 complexes 415:445 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 0 7 from intermediate 20:31 arg1 maturation 49:58 class II MHC maturation 36:58 class II MHC maturation 36:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 0 8 dep CLIP 61:64 arg1 structure 4:12 The structure 0:12 The structure of an intermediate in class II MHC maturation 0:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 2 9 theme antigenic 415:423 arg1 complexes 437:445 antigenic peptide-DR3 complexes 415:445 antigenic peptide-DR3 complexes 415:445 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 2 10 theme Previous 270:277 arg1 data 279:282 Previous data 270:282 Previous data 270:282 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 2 11 theme class 333:337 arg1 histocompatibility 342:359 class II histocompatibility 333:359 class II histocompatibility maturation 333:370 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 1 12 theme X-ray 234:238 arg1 structure 248:256 its X-ray crystal structure 230:256 its X-ray crystal structure 230:256 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 1 13 from presentation 213:224 arg1 defective 192:200 defective 192:200 defective 192:200 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 2 14 from intermediate 317:328 arg1 maturation 361:370 class II histocompatibility maturation 333:370 class II histocompatibility maturation 333:370 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 3 15 theme identical 520:528 arg1 way 509:511 a way 507:511 a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins 507:611 The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. 7477400 2 16 theme invariant 391:399 arg1 chain-DR3 401:409 invariant chain-DR3 391:409 invariant chain-DR3 391:409 Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. 7477400 1 17 theme crystal 240:246 arg1 structure 248:256 its X-ray crystal structure 230:256 its X-ray crystal structure 230:256 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 0 18 theme intermediate 20:31 arg1 structure 4:12 The structure 0:12 The structure of an intermediate in class II MHC maturation 0:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 0 19 from maturation 49:58 arg1 structure 4:12 The structure 0:12 The structure of an intermediate in class II MHC maturation 0:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 0 20 theme class 36:40 arg1 MHC 45:47 class II MHC 36:47 class II MHC maturation 36:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 1 21 from defective 192:200 arg1 presentation 213:224 antigen presentation 205:224 antigen presentation 205:224 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 1 22 attach isolated 160:167 arg1 line 187:190 a human cell line 174:190 a human cell line defective in antigen presentation 174:224 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 1 22 attach isolated 160:167 arg2 fragment 116:123 a fragment 114:123 a fragment of invariant chain called CLIP 114:154 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 1 22 attach isolated 160:167 arg2 complex 86:92 A complex 84:92 A complex between HLA-DR3 84:108 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 3 23 theme antigenic 547:555 arg1 peptides 557:564 antigenic peptides 547:564 antigenic peptides 547:564 The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. 7477400 3 24 theme CLIP 477:480 arg1 fragment 482:489 the CLIP fragment 473:489 the CLIP fragment 473:489 The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. 7477400 3 25 gly glycoproteins 599:611 arg1 glycoproteins 599:611 class II histocompatibility glycoproteins 571:611 class II histocompatibility glycoproteins 571:611 The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. 7477400 3 26 theme histocompatibility 580:597 arg1 glycoproteins 599:611 class II histocompatibility glycoproteins 571:611 class II histocompatibility glycoproteins 571:611 The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. 7477400 1 27 theme human 176:180 arg1 line 187:190 a human cell line 174:190 a human cell line defective in antigen presentation 174:224 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 4 28 theme exchange 689:696 arg1 process 698:704 an exchange process 686:704 an exchange process catalysed by DM 686:720 The structure is the substrate for the loading of antigenic peptides by an exchange process catalysed by DM. 7477400 1 29 theme cell 182:185 arg1 line 187:190 a human cell line 174:190 a human cell line defective in antigen presentation 174:224 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 3 30 theme class 571:575 arg1 glycoproteins 599:611 class II histocompatibility glycoproteins 571:611 class II histocompatibility glycoproteins 571:611 The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. 7477400 1 31 theme chain 138:142 arg1 fragment 116:123 a fragment 114:123 a fragment of invariant chain called CLIP 114:154 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 1 31 theme chain 138:142 arg1 complex 86:92 A complex 84:92 A complex between HLA-DR3 84:108 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 7477400 0 32 theme MHC 45:47 arg1 maturation 49:58 class II MHC maturation 36:58 class II MHC maturation 36:58 The structure of an intermediate in class II MHC maturation: CLIP bound to HLA-DR3. 7477400 1 33 theme defective 192:200 arg1 line 187:190 a human cell line 174:190 a human cell line defective in antigen presentation 174:224 A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. 18691975 0 0 theme alphaL 74:79 arg1 variant 99:105 a high-affinity alphaL integrin I domain variant 58:105 a high-affinity alphaL integrin I domain variant 58:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 6 1 with changes 1032:1038 arg1 penalty 1059:1065 little energy penalty 1045:1065 little energy penalty 1045:1065 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 1 2 theme cell 138:141 arg1 receptors 151:159 cell surface receptors 138:159 cell surface receptors that transduce signals bidirectionally across the plasma membrane 138:225 Integrins are cell surface receptors that transduce signals bidirectionally across the plasma membrane. 18691975 1 2 theme cell 138:141 arg1 Integrins 124:132 Integrins 124:132 Integrins 124:132 Integrins are cell surface receptors that transduce signals bidirectionally across the plasma membrane. 18691975 0 3 theme high-affinity 60:72 arg1 variant 99:105 a high-affinity alphaL integrin I domain variant 58:105 a high-affinity alphaL integrin I domain variant 58:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 1 4 theme surface 143:149 arg1 receptors 151:159 cell surface receptors 138:159 cell surface receptors that transduce signals bidirectionally across the plasma membrane 138:225 Integrins are cell surface receptors that transduce signals bidirectionally across the plasma membrane. 18691975 1 4 theme surface 143:149 arg1 Integrins 124:132 Integrins 124:132 Integrins 124:132 Integrins are cell surface receptors that transduce signals bidirectionally across the plasma membrane. 18691975 0 5 theme I 90:90 arg1 variant 99:105 a high-affinity alphaL integrin I domain variant 58:105 a high-affinity alphaL integrin I domain variant 58:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 5 6 theme unusual 903:909 arg1 mobility 911:918 an intrinsically unusual mobility 886:918 an intrinsically unusual mobility of this helix 886:932 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 7 7 theme bound 1119:1123 arg1 molecule 1134:1141 a weakly bound adhesion molecule 1110:1141 a weakly bound adhesion molecule 1110:1141 It serves as a mechanistic example of how a weakly bound adhesion molecule works in signaling. 18691975 4 8 theme N-terminal 656:665 arg1 ICAM-5 682:687 ICAM-5 682:687 ICAM-5 682:687 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 8 theme N-terminal 656:665 arg1 domains 671:677 the N-terminal two domains 652:677 the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 652:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 9 theme high-affinity 557:569 arg1 domain 573:578 an engineered high-affinity I domain 543:578 an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit 543:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 0 10 theme integrin 81:88 arg1 variant 99:105 a high-affinity alphaL integrin I domain variant 58:105 a high-affinity alphaL integrin I domain variant 58:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 2 11 theme allosteric 271:280 arg1 regulation 282:291 the allosteric regulation 267:291 the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain 267:391 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 2 11 theme allosteric 271:280 arg1 event 236:240 The key event 228:240 The key event of integrin signaling 228:262 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 4 12 theme engineered 546:555 arg1 domain 573:578 an engineered high-affinity I domain 543:578 an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit 543:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 0 13 theme variant 99:105 arg1 helix 49:53 the C-terminal helix 34:53 the C-terminal helix of a high-affinity alphaL integrin I domain variant 34:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 6 14 theme energy 1052:1057 arg1 penalty 1059:1065 little energy penalty 1045:1065 little energy penalty 1045:1065 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 4 15 theme integrin 589:596 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 3 16 theme axial 408:412 arg1 movement 414:421 A significant axial movement 394:421 A significant axial movement of the alpha7 helix 394:441 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 0 17 theme domain 92:97 arg1 variant 99:105 a high-affinity alphaL integrin I domain variant 58:105 a high-affinity alphaL integrin I domain variant 58:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 3 18 theme integrins 495:503 arg1 conformation 479:490 the open, active conformation 462:490 conformation 479:490 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 2 19 theme inserted 373:380 arg1 domain 386:391 integrin's inserted (I) domain 362:391 integrin's inserted (I) domain 362:391 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 0 20 theme unusual 3:9 arg1 mobility 22:29 An unusual allosteric mobility 0:29 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant 0:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 7 21 theme adhesion 1125:1132 arg1 molecule 1134:1141 a weakly bound adhesion molecule 1110:1141 a weakly bound adhesion molecule 1110:1141 It serves as a mechanistic example of how a weakly bound adhesion molecule works in signaling. 18691975 7 22 theme mechanistic 1083:1093 arg1 It 1068:1069 It 1068:1069 It 1068:1069 It serves as a mechanistic example of how a weakly bound adhesion molecule works in signaling. 18691975 7 22 theme mechanistic 1083:1093 arg1 example 1095:1101 a mechanistic example 1081:1101 a mechanistic example of how a weakly bound adhesion molecule works in signaling 1081:1160 It serves as a mechanistic example of how a weakly bound adhesion molecule works in signaling. 18691975 5 23 theme alpha7 768:773 arg1 helix 775:779 the alpha7 helix 764:779 the alpha7 helix 764:779 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 2 24 theme ligand-binding 305:318 arg1 site 320:323 its ligand-binding site 301:323 its ligand-binding site 301:323 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 3 25 theme significant 396:406 arg1 movement 414:421 A significant axial movement 394:421 A significant axial movement of the alpha7 helix 394:441 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 4 26 theme telencephalic 724:736 arg1 neurons 738:744 telencephalic neurons 724:744 telencephalic neurons 724:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 3 27 theme alpha7 430:435 arg1 helix 437:441 the alpha7 helix 426:441 the alpha7 helix 426:441 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 0 28 theme allosteric 11:20 arg1 mobility 22:29 An unusual allosteric mobility 0:29 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant 0:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 5 29 theme upside-down 838:848 arg1 orientation 850:860 an upside-down orientation 835:860 an upside-down orientation in the crystals 835:876 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 4 30 theme LFA-1 615:619 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 3 31 theme helix 437:441 arg1 movement 414:421 A significant axial movement 394:421 A significant axial movement of the alpha7 helix 394:441 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 3 32 theme open 466:469 arg1 conformation 479:490 the open, active conformation 462:490 conformation 479:490 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 4 33 theme crystal 522:528 arg1 structure 530:538 the crystal structure 518:538 the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 518:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 6 34 theme little 1045:1050 arg1 penalty 1059:1065 little energy penalty 1045:1065 little energy penalty 1045:1065 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 2 35 theme key 232:234 arg1 regulation 282:291 the allosteric regulation 267:291 the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain 267:391 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 2 35 theme key 232:234 arg1 event 236:240 The key event 228:240 The key event of integrin signaling 228:262 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 6 36 theme alpha7 970:975 arg1 helix 977:981 the alpha7 helix 966:981 the alpha7 helix 966:981 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 4 37 theme L 604:604 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 38 theme beta 606:609 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 5 39 theme I 823:823 arg1 domain 825:830 a neighboring I domain 809:830 a neighboring I domain 809:830 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 1 40 theme plasma 211:216 arg1 membrane 218:225 the plasma membrane 207:225 the plasma membrane 207:225 Integrins are cell surface receptors that transduce signals bidirectionally across the plasma membrane. 18691975 0 41 theme helix 49:53 arg1 mobility 22:29 An unusual allosteric mobility 0:29 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant 0:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 6 42 theme conformational 1017:1030 arg1 changes 1032:1038 integrin's large-scale conformational changes 994:1038 integrin's large-scale conformational changes with little energy penalty 994:1065 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 5 43 theme neighboring 811:821 arg1 domain 825:830 a neighboring I domain 809:830 a neighboring I domain 809:830 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 0 44 theme C-terminal 38:47 arg1 helix 49:53 the C-terminal helix 34:53 the C-terminal helix of a high-affinity alphaL integrin I domain variant 34:105 An unusual allosteric mobility of the C-terminal helix of a high-affinity alphaL integrin I domain variant bound to ICAM-5. 18691975 2 45 theme C-terminal 333:342 arg1 alpha7 351:356 alpha7 351:356 alpha7 351:356 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 2 45 theme C-terminal 333:342 arg1 helix 344:348 the C-terminal helix 329:348 the C-terminal helix (alpha7) of integrin's inserted (I) domain 329:391 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 5 46 from orientation 850:860 arg1 crystals 869:876 the crystals 865:876 the crystals 865:876 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 4 47 theme adhesion 693:700 arg1 molecule 702:709 an adhesion molecule 690:709 an adhesion molecule expressed in telencephalic neurons 690:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 47 theme adhesion 693:700 arg1 ICAM-5 682:687 ICAM-5 682:687 ICAM-5 682:687 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 2 48 theme signaling 254:262 arg1 regulation 282:291 the allosteric regulation 267:291 the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain 267:391 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 2 48 theme signaling 254:262 arg1 event 236:240 The key event 228:240 The key event of integrin signaling 228:262 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 5 49 theme helix 928:932 arg1 mobility 911:918 an intrinsically unusual mobility 886:918 an intrinsically unusual mobility of this helix 886:932 The finding that the alpha7 helix swings out and inserts into a neighboring I domain in an upside-down orientation in the crystals implies an intrinsically unusual mobility of this helix. 18691975 4 50 theme domain 573:578 arg1 structure 530:538 the crystal structure 518:538 the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 518:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 51 theme ICAM-5 682:687 arg1 ICAM-5 682:687 ICAM-5 682:687 ICAM-5 682:687 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 51 theme ICAM-5 682:687 arg1 domains 671:677 the N-terminal two domains 652:677 the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 652:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 2 52 theme integrin 245:252 arg1 signaling 254:262 integrin signaling 245:262 integrin signaling 245:262 The key event of integrin signaling is the allosteric regulation between its ligand-binding site and the C-terminal helix (alpha7) of integrin's inserted (I) domain. 18691975 6 53 theme large-scale 1005:1015 arg1 changes 1032:1038 integrin's large-scale conformational changes 994:1038 integrin's large-scale conformational changes with little energy penalty 994:1065 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 4 54 from subunit 628:634 arg1 domain 573:578 an engineered high-affinity I domain 543:578 an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit 543:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 54 from subunit 628:634 arg1 structure 530:538 the crystal structure 518:538 the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 518:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 55 theme I 571:571 arg1 domain 573:578 an engineered high-affinity I domain 543:578 an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit 543:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 6 56 theme remarkable 940:949 arg1 feature 951:957 This remarkable feature 935:957 This remarkable feature 935:957 This remarkable feature allows the alpha7 helix to trigger integrin's large-scale conformational changes with little energy penalty. 18691975 4 57 theme alpha 598:602 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 3 58 theme active 472:477 arg1 conformation 479:490 the open, active conformation 462:490 conformation 479:490 A significant axial movement of the alpha7 helix is associated with the open, active conformation of integrins. 18691975 4 59 with complex 639:645 arg1 ICAM-5 682:687 ICAM-5 682:687 ICAM-5 682:687 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 59 with complex 639:645 arg1 domains 671:677 the N-terminal two domains 652:677 the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 652:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 60 from structure 530:538 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 60 from structure 530:538 arg1 complex 639:645 complex 639:645 complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons 639:744 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 18691975 4 61 theme alpha 622:626 arg1 subunit 628:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 the integrin alpha(L)beta(2) (LFA-1) alpha subunit 585:634 We describe the crystal structure of an engineered high-affinity I domain from the integrin alpha(L)beta(2) (LFA-1) alpha subunit in complex with the N-terminal two domains of ICAM-5, an adhesion molecule expressed in telencephalic neurons. 12408961 0 0 theme putative 73:80 arg1 glycoproteins 91:103 putative secreted glycoproteins 73:103 putative secreted glycoproteins 73:103 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 0 0 theme putative 73:80 arg1 members 45:51 novel members 39:51 novel members of the CREG family 39:70 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 6 1 theme N 944:944 arg1 sequences 994:1002 signal sequences 987:1002 signal sequences 987:1002 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 6 1 theme N 944:944 arg1 termini 946:952 the N termini 940:952 the N termini of both proteins 940:969 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 7 2 theme neuronal 1086:1093 arg1 molecules 1109:1117 novel neuronal extracellular molecules 1080:1117 novel neuronal extracellular molecules 1080:1117 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 1 3 theme pluripotent 252:262 arg1 cells 269:273 pluripotent stem cells 252:273 pluripotent stem cells 252:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 7 4 theme putative 1037:1044 arg1 human 1011:1015 human 1011:1015 human 1011:1015 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 7 4 theme putative 1037:1044 arg1 glycoproteins 1055:1067 putative secreted glycoproteins 1037:1067 putative secreted glycoproteins 1037:1067 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 7 4 theme putative 1037:1044 arg1 mouse 1021:1025 mouse 1021:1025 mouse 1021:1025 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 6 5 theme sequences 876:884 arg1 deletion 849:856 deletion 849:856 deletion of amino-terminal sequences 849:884 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 7 6 theme secreted 1046:1053 arg1 human 1011:1015 human 1011:1015 human 1011:1015 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 7 6 theme secreted 1046:1053 arg1 glycoproteins 1055:1067 putative secreted glycoproteins 1037:1067 putative secreted glycoproteins 1037:1067 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 7 6 theme secreted 1046:1053 arg1 mouse 1021:1025 mouse 1021:1025 mouse 1021:1025 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 1 7 theme E1A-stimulated 164:177 arg1 genes 179:183 E1A-stimulated genes 164:183 E1A-stimulated genes 164:183 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 8 theme stem 264:267 arg1 cells 269:273 pluripotent stem cells 252:273 pluripotent stem cells 252:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 4 9 theme CREG2 496:500 arg1 transcription 512:524 specific CREG2 and Creg2 transcription 487:524 specific CREG2 and Creg2 transcription in brain-in the case of CREG2 487:554 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 1 10 theme cells 269:273 arg1 differentiation 233:247 differentiation 233:247 differentiation of pluripotent stem cells 233:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 0 11 theme secreted 82:89 arg1 glycoproteins 91:103 putative secreted glycoproteins 73:103 putative secreted glycoproteins 73:103 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 0 11 theme secreted 82:89 arg1 members 45:51 novel members 39:51 novel members of the CREG family 39:70 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 4 12 theme Northern 452:459 arg1 analyses 466:473 Northern blot analyses 452:473 Northern blot analyses 452:473 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 7 13 theme novel 1080:1084 arg1 molecules 1109:1117 novel neuronal extracellular molecules 1080:1117 novel neuronal extracellular molecules 1080:1117 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 5 14 from EGFP 659:662 arg1 cells 674:678 NIH3T3 cells 667:678 NIH3T3 cells 667:678 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 7 15 dep human 1011:1015 arg1 CREG2 1027:1031 CREG2 1027:1031 CREG2 1027:1031 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 3 16 dep human 378:382 arg1 sequences 402:410 protein sequences 394:410 protein sequences 394:410 The predicted human and mouse protein sequences exhibit 35% identity with CREG protein. 12408961 4 17 theme brain 592:596 arg1 system 578:583 the limbic system 567:583 the limbic system of the brain 567:596 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 1 18 theme genes 179:183 arg1 CREG 186:189 CREG 186:189 CREG 186:189 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 18 theme genes 179:183 arg1 repressor 151:159 The cellular repressor 138:159 The cellular repressor of E1A-stimulated genes 138:183 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 18 theme genes 179:183 arg1 glycoprotein 206:217 a secreted glycoprotein 195:217 a secreted glycoprotein that enhances differentiation of pluripotent stem cells 195:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 5 19 theme reticulum 756:764 arg1 localization 776:787 implicit endoplasmic reticulum and Golgi localization 735:787 implicit endoplasmic reticulum and Golgi localization 735:787 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 5 20 dep Both 599:602 arg1 mouse 604:608 mouse 604:608 mouse 604:608 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 5 21 from terminus 647:654 arg1 cells 674:678 NIH3T3 cells 667:678 NIH3T3 cells 667:678 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 3 22 theme % 422:422 arg1 identity 424:431 35% identity 420:431 35% identity with CREG protein 420:449 The predicted human and mouse protein sequences exhibit 35% identity with CREG protein. 12408961 5 23 dep CREG2 620:624 arg1 Both 599:602 Both 599:602 Both 599:602 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 5 23 dep CREG2 620:624 arg1 human 614:618 human 614:618 human 614:618 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 4 24 theme blot 461:464 arg1 analyses 466:473 Northern blot analyses 452:473 Northern blot analyses 452:473 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 1 25 gly glycoprotein 206:217 arg1 repressor 151:159 The cellular repressor 138:159 The cellular repressor of E1A-stimulated genes 138:183 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 25 gly glycoprotein 206:217 arg1 glycoprotein 206:217 a secreted glycoprotein 195:217 a secreted glycoprotein that enhances differentiation of pluripotent stem cells 195:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 5 26 theme perinuclear 696:706 arg1 region 708:713 the perinuclear region 692:713 the perinuclear region 692:713 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 7 27 gly glycoproteins 1055:1067 arg1 human 1011:1015 human 1011:1015 human 1011:1015 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 7 27 gly glycoproteins 1055:1067 arg1 glycoproteins 1055:1067 putative secreted glycoproteins 1037:1067 putative secreted glycoproteins 1037:1067 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 7 27 gly glycoproteins 1055:1067 arg1 mouse 1021:1025 mouse 1021:1025 mouse 1021:1025 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 5 28 theme Golgi 770:774 arg1 localization 776:787 implicit endoplasmic reticulum and Golgi localization 735:787 implicit endoplasmic reticulum and Golgi localization 735:787 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 0 29 gly glycoproteins 91:103 arg1 glycoproteins 91:103 putative secreted glycoproteins 73:103 putative secreted glycoproteins 73:103 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 0 29 gly glycoproteins 91:103 arg1 members 45:51 novel members 39:51 novel members of the CREG family 39:70 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 7 30 theme extracellular 1095:1107 arg1 molecules 1109:1117 novel neuronal extracellular molecules 1080:1117 novel neuronal extracellular molecules 1080:1117 Thus, human and mouse CREG2 are putative secreted glycoproteins and may be novel neuronal extracellular molecules. 12408961 3 31 with identity 424:431 arg1 protein 443:449 CREG protein 438:449 CREG protein 438:449 The predicted human and mouse protein sequences exhibit 35% identity with CREG protein. 12408961 6 32 from N-glycosylated 816:829 arg1 cells 839:843 HeLa cells 834:843 HeLa cells 834:843 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 5 33 theme endoplasmic 744:754 arg1 reticulum 756:764 implicit endoplasmic reticulum 735:764 implicit endoplasmic reticulum 735:764 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 3 34 theme CREG 438:441 arg1 protein 443:449 CREG protein 438:449 CREG protein 438:449 The predicted human and mouse protein sequences exhibit 35% identity with CREG protein. 12408961 4 35 theme CREG2 550:554 arg1 case 542:545 brain-in the case 529:545 brain-in the case of CREG2 529:554 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 5 36 theme EGFP 659:662 arg1 terminus 647:654 the carboxy terminus 635:654 the carboxy terminus of EGFP in NIH3T3 cells 635:678 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 6 37 theme HeLa 834:837 arg1 cells 839:843 HeLa cells 834:843 HeLa cells 834:843 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 2 38 theme novel 295:299 arg1 members 301:307 two novel members 291:307 two novel members of the CREG family 291:326 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 38 theme novel 295:299 arg1 cDNAs 357:361 mouse Creg2 cDNAs 345:361 mouse Creg2 cDNAs 345:361 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 38 theme novel 295:299 arg1 CREG2 335:339 human CREG2 329:339 human CREG2 329:339 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 1 39 theme cellular 142:149 arg1 CREG 186:189 CREG 186:189 CREG 186:189 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 39 theme cellular 142:149 arg1 repressor 151:159 The cellular repressor 138:159 The cellular repressor of E1A-stimulated genes 138:183 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 39 theme cellular 142:149 arg1 glycoprotein 206:217 a secreted glycoprotein 195:217 a secreted glycoprotein that enhances differentiation of pluripotent stem cells 195:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 0 40 theme members 45:51 arg1 characterization 19:34 characterization 19:34 characterization 19:34 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 0 40 theme members 45:51 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 6 41 gly N-glycosylated 816:829 arg1 cells 839:843 HeLa cells 834:843 HeLa cells 834:843 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 6 41 gly N-glycosylated 816:829 arg1 CREG2 806:810 Human and mouse CREG2 790:810 Human and mouse CREG2 790:810 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 6 41 gly N-glycosylated 816:829 arg2 CREG2 806:810 Human and mouse CREG2 790:810 Human and mouse CREG2 790:810 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 1 42 theme secreted 197:204 arg1 repressor 151:159 The cellular repressor 138:159 The cellular repressor of E1A-stimulated genes 138:183 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 1 42 theme secreted 197:204 arg1 glycoprotein 206:217 a secreted glycoprotein 195:217 a secreted glycoprotein that enhances differentiation of pluripotent stem cells 195:273 The cellular repressor of E1A-stimulated genes, CREG, is a secreted glycoprotein that enhances differentiation of pluripotent stem cells. 12408961 0 43 theme novel 39:43 arg1 members 45:51 novel members 39:51 novel members of the CREG family 39:70 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 0 43 theme novel 39:43 arg1 glycoproteins 91:103 putative secreted glycoproteins 73:103 putative secreted glycoproteins 73:103 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 3 44 theme protein 394:400 arg1 sequences 402:410 protein sequences 394:410 protein sequences 394:410 The predicted human and mouse protein sequences exhibit 35% identity with CREG protein. 12408961 5 45 from cells 674:678 arg1 terminus 647:654 the carboxy terminus 635:654 the carboxy terminus of EGFP in NIH3T3 cells 635:678 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 6 46 theme amino-terminal 861:874 arg1 sequences 876:884 amino-terminal sequences 861:884 amino-terminal sequences 861:884 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 4 47 from transcription 512:524 arg1 case 542:545 brain-in the case 529:545 brain-in the case of CREG2 529:554 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 4 48 theme Creg2 506:510 arg1 transcription 512:524 specific CREG2 and Creg2 transcription 487:524 specific CREG2 and Creg2 transcription in brain-in the case of CREG2 487:554 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 3 49 theme 35 420:421 arg1 % 422:422 % 422:422 % 422:422 The predicted human and mouse protein sequences exhibit 35% identity with CREG protein. 12408961 2 50 theme human 329:333 arg1 members 301:307 two novel members 291:307 two novel members of the CREG family 291:326 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 50 theme human 329:333 arg1 CREG2 335:339 human CREG2 329:339 human CREG2 329:339 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 5 51 theme NIH3T3 667:672 arg1 cells 674:678 NIH3T3 cells 667:678 NIH3T3 cells 667:678 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 4 52 theme brain-in 529:536 arg1 case 542:545 brain-in the case 529:545 brain-in the case of CREG2 529:554 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 5 53 theme carboxy 639:645 arg1 terminus 647:654 the carboxy terminus 635:654 the carboxy terminus of EGFP in NIH3T3 cells 635:678 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 6 54 theme mouse 800:804 arg1 CREG2 806:810 Human and mouse CREG2 790:810 Human and mouse CREG2 790:810 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 2 55 theme family 321:326 arg1 members 301:307 two novel members 291:307 two novel members of the CREG family 291:326 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 55 theme family 321:326 arg1 cDNAs 357:361 mouse Creg2 cDNAs 345:361 mouse Creg2 cDNAs 345:361 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 55 theme family 321:326 arg1 CREG2 335:339 human CREG2 329:339 human CREG2 329:339 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 6 56 theme proteins 962:969 arg1 sequences 994:1002 signal sequences 987:1002 signal sequences 987:1002 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 6 56 theme proteins 962:969 arg1 termini 946:952 the N termini 940:952 the N termini of both proteins 940:969 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 2 57 theme CREG 316:319 arg1 family 321:326 the CREG family 312:326 the CREG family 312:326 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 4 58 theme limbic 571:576 arg1 system 578:583 the limbic system 567:583 the limbic system of the brain 567:596 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 0 59 theme family 65:70 arg1 members 45:51 novel members 39:51 novel members of the CREG family 39:70 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 0 59 theme family 65:70 arg1 glycoproteins 91:103 putative secreted glycoproteins 73:103 putative secreted glycoproteins 73:103 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 6 60 theme Human 790:794 arg1 CREG2 806:810 Human and mouse CREG2 790:810 Human and mouse CREG2 790:810 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 2 61 theme Creg2 351:355 arg1 members 301:307 two novel members 291:307 two novel members of the CREG family 291:326 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 61 theme Creg2 351:355 arg1 cDNAs 357:361 mouse Creg2 cDNAs 345:361 mouse Creg2 cDNAs 345:361 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 6 62 theme signal 987:992 arg1 termini 946:952 the N termini 940:952 the N termini of both proteins 940:969 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 6 62 theme signal 987:992 arg1 sequences 994:1002 signal sequences 987:1002 signal sequences 987:1002 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 12408961 0 63 theme CREG 60:63 arg1 family 65:70 the CREG family 56:70 the CREG family 56:70 Identification and characterization of novel members of the CREG family, putative secreted glycoproteins expressed specifically in brain. 12408961 4 64 theme specific 487:494 arg1 transcription 512:524 specific CREG2 and Creg2 transcription 487:524 specific CREG2 and Creg2 transcription in brain-in the case of CREG2 487:554 Northern blot analyses demonstrate specific CREG2 and Creg2 transcription in brain-in the case of CREG2, mainly in the limbic system of the brain. 12408961 5 65 theme implicit 735:742 arg1 reticulum 756:764 implicit endoplasmic reticulum 735:764 implicit endoplasmic reticulum 735:764 Both mouse and human CREG2 fused to the carboxy terminus of EGFP in NIH3T3 cells localize to the perinuclear region, which demonstrates implicit endoplasmic reticulum and Golgi localization. 12408961 2 66 theme mouse 345:349 arg1 members 301:307 two novel members 291:307 two novel members of the CREG family 291:326 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 2 66 theme mouse 345:349 arg1 cDNAs 357:361 mouse Creg2 cDNAs 345:361 mouse Creg2 cDNAs 345:361 Here we report two novel members of the CREG family, human CREG2 and mouse Creg2 cDNAs. 12408961 6 67 from cells 839:843 arg1 N-glycosylated 816:829 N-glycosylated 816:829 N-glycosylated 816:829 Human and mouse CREG2 are N-glycosylated in HeLa cells and deletion of amino-terminal sequences completely abolishes N-glycosylation, indicating that the N termini of both proteins may function as signal sequences. 15687489 0 0 theme enzyme 84:89 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human pFGE 0:30 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 0 0 theme enzyme 84:89 arg1 paralog 37:43 the paralog 33:43 the paralog of the Calpha-formylglycine-generating enzyme 33:89 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 12 1 theme binding 1434:1440 arg1 site 1442:1445 The putative peptide binding site 1413:1445 The putative peptide binding site 1413:1445 The putative peptide binding site is buried between the monomers, indicating a biological significance of the dimer. 15687489 9 2 theme asymmetric 1154:1163 arg1 partitioning 1165:1176 an asymmetric partitioning 1151:1176 an asymmetric partitioning of secondary structure elements 1151:1208 It is characterized by an asymmetric partitioning of secondary structure elements and is stabilized by two calcium cations. 15687489 7 3 theme human 924:928 arg1 pFGE 930:933 the glycosylated human pFGE 907:933 the glycosylated human pFGE 907:933 We have crystallized the glycosylated human pFGE and determined its crystal structure at a resolution of 1.86 A. 15687489 5 4 link sulfatase-derived 595:611 arg1 peptide 613:619 a sulfatase-derived peptide 593:619 a sulfatase-derived peptide bearing the FGE recognition motif 593:653 pFGE binds a sulfatase-derived peptide bearing the FGE recognition motif, but it lacks formylglycine-generating activity. 15687489 0 5 theme Calpha-formylglycine-generating 52:82 arg1 enzyme 84:89 the Calpha-formylglycine-generating enzyme 48:89 the Calpha-formylglycine-generating enzyme 48:89 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 6 6 theme enzyme 832:837 arg1 domain 839:844 a formylglycine-generating enzyme domain 805:844 a formylglycine-generating enzyme domain of unknown three-dimensional structure 805:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 6 theme enzyme 832:837 arg1 domain 797:802 the DUF323 domain 786:802 the DUF323 domain 786:802 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 6 theme enzyme 832:837 arg1 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 7 theme structure 875:883 arg1 domain 839:844 a formylglycine-generating enzyme domain 805:844 a formylglycine-generating enzyme domain of unknown three-dimensional structure 805:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 7 theme structure 875:883 arg1 domain 797:802 the DUF323 domain 786:802 the DUF323 domain 786:802 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 7 theme structure 875:883 arg1 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 4 8 theme FGE 576:578 arg1 paralog 565:571 paralog 565:571 paralog of FGE 565:578 Using sequence comparisons, a protein homologous to FGE was found and denoted pFGE (paralog of FGE). 15687489 6 9 contain containing 775:784 arg2 domain 839:844 a formylglycine-generating enzyme domain 805:844 a formylglycine-generating enzyme domain of unknown three-dimensional structure 805:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 9 contain containing 775:784 arg2 domain 797:802 the DUF323 domain 786:802 the DUF323 domain 786:802 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 9 contain containing 775:784 arg2 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 9 contain containing 775:784 arg1 proteins 766:773 pro- and eukaryotic proteins 746:773 pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure 746:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 7 10 theme A 996:996 arg1 resolution 977:986 a resolution 975:986 a resolution of 1.86 A 975:996 We have crystallized the glycosylated human pFGE and determined its crystal structure at a resolution of 1.86 A. 15687489 2 11 from disorder 337:344 arg1 humans 349:354 humans 349:354 humans 349:354 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 11 12 contain contains 1391:1398 arg2 homodimer 1402:1410 a homodimer 1400:1410 a homodimer 1400:1410 The asymmetric unit of the pFGE crystal contains a homodimer. 15687489 11 12 contain contains 1391:1398 arg1 unit 1366:1369 The asymmetric unit 1351:1369 The asymmetric unit of the pFGE crystal 1351:1389 The asymmetric unit of the pFGE crystal contains a homodimer. 15687489 13 13 theme pFGE 1571:1574 arg1 capability 1557:1566 the capability 1553:1566 the capability of pFGE to form a heterodimer with FGE 1553:1605 The structure suggests the capability of pFGE to form a heterodimer with FGE. 15687489 12 14 theme biological 1492:1501 arg1 significance 1503:1514 a biological significance 1490:1514 a biological significance of the dimer 1490:1527 The putative peptide binding site is buried between the monomers, indicating a biological significance of the dimer. 15687489 10 15 theme polypeptide 1325:1335 arg1 site 1345:1348 the sulfatase polypeptide binding site 1311:1348 the sulfatase polypeptide binding site 1311:1348 A deep cleft on the surface of pFGE most likely represents the sulfatase polypeptide binding site. 15687489 5 16 theme sulfatase-derived 595:611 arg1 peptide 613:619 a sulfatase-derived peptide 593:619 a sulfatase-derived peptide bearing the FGE recognition motif 593:653 pFGE binds a sulfatase-derived peptide bearing the FGE recognition motif, but it lacks formylglycine-generating activity. 15687489 2 17 theme residue 295:301 arg1 formation 257:265 post-translational formation 238:265 post-translational formation of the Calpha-formylglycine residue 238:301 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 12 18 theme dimer 1523:1527 arg1 significance 1503:1514 a biological significance 1490:1514 a biological significance of the dimer 1490:1527 The putative peptide binding site is buried between the monomers, indicating a biological significance of the dimer. 15687489 11 19 theme crystal 1383:1389 arg1 unit 1366:1369 The asymmetric unit 1351:1369 The asymmetric unit of the pFGE crystal 1351:1389 The asymmetric unit of the pFGE crystal contains a homodimer. 15687489 4 20 theme sequence 487:494 arg1 comparisons 496:506 sequence comparisons 487:506 sequence comparisons 487:506 Using sequence comparisons, a protein homologous to FGE was found and denoted pFGE (paralog of FGE). 15687489 9 21 theme structure 1191:1199 arg1 elements 1201:1208 secondary structure elements 1181:1208 secondary structure elements 1181:1208 It is characterized by an asymmetric partitioning of secondary structure elements and is stabilized by two calcium cations. 15687489 2 22 theme Calpha-formylglycine 274:293 arg1 residue 295:301 the Calpha-formylglycine residue 270:301 the Calpha-formylglycine residue 270:301 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 12 23 theme peptide 1426:1432 arg1 site 1442:1445 The putative peptide binding site 1413:1445 The putative peptide binding site 1413:1445 The putative peptide binding site is buried between the monomers, indicating a biological significance of the dimer. 15687489 8 24 theme FGE 1055:1057 arg1 fold 1059:1062 the FGE fold 1051:1062 the FGE fold 1051:1062 The structure reveals a novel fold, which we denote the FGE fold and which therefore serves as a paradigm for the DUF323 domain. 15687489 8 25 theme novel 1023:1027 arg1 fold 1029:1032 a novel fold 1021:1032 a novel fold 1021:1032 The structure reveals a novel fold, which we denote the FGE fold and which therefore serves as a paradigm for the DUF323 domain. 15687489 2 26 from defect 228:233 arg1 formation 257:265 post-translational formation 238:265 post-translational formation of the Calpha-formylglycine residue 238:301 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 4 27 dep pFGE 559:562 arg1 paralog 565:571 paralog 565:571 paralog of FGE 565:578 Using sequence comparisons, a protein homologous to FGE was found and denoted pFGE (paralog of FGE). 15687489 5 28 theme FGE 633:635 arg1 motif 649:653 the FGE recognition motif 629:653 the FGE recognition motif 629:653 pFGE binds a sulfatase-derived peptide bearing the FGE recognition motif, but it lacks formylglycine-generating activity. 15687489 6 29 theme DUF323 790:795 arg1 domain 839:844 a formylglycine-generating enzyme domain 805:844 a formylglycine-generating enzyme domain of unknown three-dimensional structure 805:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 29 theme DUF323 790:795 arg1 domain 797:802 the DUF323 domain 786:802 the DUF323 domain 786:802 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 29 theme DUF323 790:795 arg1 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 2 30 theme storage 329:335 arg1 disorder 337:344 a severe lysosomal storage disorder 310:344 a severe lysosomal storage disorder in humans 310:354 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 1 31 theme unique 166:171 arg1 residue 194:200 a unique Calpha-formylglycine residue 164:200 a unique Calpha-formylglycine residue in their active site 164:221 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 0 32 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human pFGE 0:30 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 0 32 theme Crystal 0:6 arg1 paralog 37:43 the paralog 33:43 the paralog of the Calpha-formylglycine-generating enzyme 33:89 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 6 33 theme formylglycine-generating 807:830 arg1 domain 839:844 a formylglycine-generating enzyme domain 805:844 a formylglycine-generating enzyme domain of unknown three-dimensional structure 805:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 33 theme formylglycine-generating 807:830 arg1 domain 797:802 the DUF323 domain 786:802 the DUF323 domain 786:802 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 33 theme formylglycine-generating 807:830 arg1 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 5 34 theme recognition 637:647 arg1 motif 649:653 the FGE recognition motif 629:653 the FGE recognition motif 629:653 pFGE binds a sulfatase-derived peptide bearing the FGE recognition motif, but it lacks formylglycine-generating activity. 15687489 7 35 theme crystal 954:960 arg1 structure 962:970 its crystal structure 950:970 its crystal structure 950:970 We have crystallized the glycosylated human pFGE and determined its crystal structure at a resolution of 1.86 A. 15687489 2 36 theme lysosomal 319:327 arg1 disorder 337:344 a severe lysosomal storage disorder 310:344 a severe lysosomal storage disorder in humans 310:354 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 1 37 theme Calpha-formylglycine 173:192 arg1 residue 194:200 a unique Calpha-formylglycine residue 164:200 a unique Calpha-formylglycine residue in their active site 164:221 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 3 38 theme formylglycine-generating 372:395 arg1 FGE 367:369 FGE 367:369 FGE (formylglycine-generating enzyme) 367:403 Recently, FGE (formylglycine-generating enzyme) has been identified as the protein required for this specific modification. 15687489 3 38 theme formylglycine-generating 372:395 arg1 enzyme 397:402 formylglycine-generating enzyme 372:402 formylglycine-generating enzyme 372:402 Recently, FGE (formylglycine-generating enzyme) has been identified as the protein required for this specific modification. 15687489 2 39 theme severe 312:317 arg1 disorder 337:344 a severe lysosomal storage disorder 310:344 a severe lysosomal storage disorder in humans 310:354 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 10 40 theme sulfatase 1315:1323 arg1 site 1345:1348 the sulfatase polypeptide binding site 1311:1348 the sulfatase polypeptide binding site 1311:1348 A deep cleft on the surface of pFGE most likely represents the sulfatase polypeptide binding site. 15687489 13 41 with heterodimer 1586:1596 arg1 FGE 1603:1605 FGE 1603:1605 FGE 1603:1605 The structure suggests the capability of pFGE to form a heterodimer with FGE. 15687489 7 42 gly glycosylated 911:922 arg1 pFGE 930:933 the glycosylated human pFGE 907:933 the glycosylated human pFGE 907:933 We have crystallized the glycosylated human pFGE and determined its crystal structure at a resolution of 1.86 A. 15687489 10 43 theme pFGE 1283:1286 arg1 surface 1272:1278 the surface 1268:1278 the surface of pFGE 1268:1286 A deep cleft on the surface of pFGE most likely represents the sulfatase polypeptide binding site. 15687489 3 44 theme specific 458:465 arg1 modification 467:478 this specific modification 453:478 this specific modification 453:478 Recently, FGE (formylglycine-generating enzyme) has been identified as the protein required for this specific modification. 15687489 0 45 theme pFGE 27:30 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human pFGE 0:30 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 0 45 theme pFGE 27:30 arg1 paralog 37:43 the paralog 33:43 the paralog of the Calpha-formylglycine-generating enzyme 33:89 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 11 46 theme asymmetric 1355:1364 arg1 unit 1366:1369 The asymmetric unit 1351:1369 The asymmetric unit of the pFGE crystal 1351:1389 The asymmetric unit of the pFGE crystal contains a homodimer. 15687489 9 47 theme calcium 1235:1241 arg1 cations 1243:1249 two calcium cations 1231:1249 two calcium cations 1231:1249 It is characterized by an asymmetric partitioning of secondary structure elements and is stabilized by two calcium cations. 15687489 0 48 theme human 21:25 arg1 pFGE 27:30 human pFGE 21:30 human pFGE 21:30 Crystal structure of human pFGE, the paralog of the Calpha-formylglycine-generating enzyme. 15687489 6 49 theme proteins 766:773 arg1 family 736:741 a large family 728:741 a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure 728:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 1 50 theme active 211:216 arg1 site 218:221 their active site 205:221 their active site 205:221 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 9 51 theme secondary 1181:1189 arg1 elements 1201:1208 secondary structure elements 1181:1208 secondary structure elements 1181:1208 It is characterized by an asymmetric partitioning of secondary structure elements and is stabilized by two calcium cations. 15687489 1 52 contain possess 156:162 arg1 sulfatases 138:147 sulfatases 138:147 sulfatases 138:147 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 1 52 contain possess 156:162 arg2 residue 194:200 a unique Calpha-formylglycine residue 164:200 a unique Calpha-formylglycine residue in their active site 164:221 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 6 53 theme eukaryotic 755:764 arg1 proteins 766:773 pro- and eukaryotic proteins 746:773 pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure 746:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 1 54 theme sulfate 107:113 arg1 esters 115:120 sulfate esters 107:120 sulfate esters 107:120 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 5 55 theme formylglycine-generating 669:692 arg1 activity 694:701 formylglycine-generating activity 669:701 formylglycine-generating activity 669:701 pFGE binds a sulfatase-derived peptide bearing the FGE recognition motif, but it lacks formylglycine-generating activity. 15687489 6 56 theme three-dimensional 857:873 arg1 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 1 57 from residue 194:200 arg1 site 218:221 their active site 205:221 their active site 205:221 In eukaryotes, sulfate esters are degraded by sulfatases, which possess a unique Calpha-formylglycine residue in their active site. 15687489 8 58 theme DUF323 1113:1118 arg1 domain 1120:1125 the DUF323 domain 1109:1125 the DUF323 domain 1109:1125 The structure reveals a novel fold, which we denote the FGE fold and which therefore serves as a paradigm for the DUF323 domain. 15687489 6 59 theme unknown 849:855 arg1 structure 875:883 unknown three-dimensional structure 849:883 unknown three-dimensional structure 849:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 6 60 theme pro- 746:749 arg1 proteins 766:773 pro- and eukaryotic proteins 746:773 pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure 746:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 9 61 theme elements 1201:1208 arg1 partitioning 1165:1176 an asymmetric partitioning 1151:1176 an asymmetric partitioning of secondary structure elements 1151:1208 It is characterized by an asymmetric partitioning of secondary structure elements and is stabilized by two calcium cations. 15687489 10 62 from cleft 1259:1263 arg1 surface 1272:1278 the surface 1268:1278 the surface of pFGE 1268:1286 A deep cleft on the surface of pFGE most likely represents the sulfatase polypeptide binding site. 15687489 10 63 theme binding 1337:1343 arg1 site 1345:1348 the sulfatase polypeptide binding site 1311:1348 the sulfatase polypeptide binding site 1311:1348 A deep cleft on the surface of pFGE most likely represents the sulfatase polypeptide binding site. 15687489 10 64 theme deep 1254:1257 arg1 cleft 1259:1263 A deep cleft 1252:1263 A deep cleft on the surface of pFGE 1252:1286 A deep cleft on the surface of pFGE most likely represents the sulfatase polypeptide binding site. 15687489 2 65 theme post-translational 238:255 arg1 formation 257:265 post-translational formation 238:265 post-translational formation of the Calpha-formylglycine residue 238:301 The defect in post-translational formation of the Calpha-formylglycine residue causes a severe lysosomal storage disorder in humans. 15687489 11 66 theme pFGE 1378:1381 arg1 crystal 1383:1389 the pFGE crystal 1374:1389 the pFGE crystal 1374:1389 The asymmetric unit of the pFGE crystal contains a homodimer. 15687489 4 67 theme homologous 519:528 arg1 protein 511:517 a protein 509:517 a protein homologous to FGE 509:535 Using sequence comparisons, a protein homologous to FGE was found and denoted pFGE (paralog of FGE). 15687489 12 68 theme putative 1417:1424 arg1 site 1442:1445 The putative peptide binding site 1413:1445 The putative peptide binding site 1413:1445 The putative peptide binding site is buried between the monomers, indicating a biological significance of the dimer. 15687489 6 69 theme large 730:734 arg1 family 736:741 a large family 728:741 a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure 728:883 Both proteins belong to a large family of pro- and eukaryotic proteins containing the DUF323 domain, a formylglycine-generating enzyme domain of unknown three-dimensional structure. 15687489 7 70 theme glycosylated 911:922 arg1 pFGE 930:933 the glycosylated human pFGE 907:933 the glycosylated human pFGE 907:933 We have crystallized the glycosylated human pFGE and determined its crystal structure at a resolution of 1.86 A. 27049939 6 0 theme allosteric 931:940 arg1 site 942:945 the allosteric site 927:945 the allosteric site 927:945 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 7 1 theme drug 1195:1198 arg1 design 1200:1205 future drug design 1188:1205 future drug design 1188:1205 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 7 2 from mechanism 1117:1125 arg1 SERT 1155:1158 SERT 1155:1158 SERT 1155:1158 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 4 3 theme transmembrane 615:627 arg1 helices 629:635 transmembrane helices 1, 3, 6, 8 and 10 615:653 transmembrane helices 1, 3, 6, 8 and 10 615:653 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 1 4 theme chloride-dependent 159:176 arg1 reuptake 178:185 the sodium- and chloride-dependent reuptake 143:185 the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons 143:230 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 27049939 3 5 theme 3.15 Å 424:429 arg1 resolution 431:440 3.15 Å resolution 424:440 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine 424:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 6 6 theme ligand 966:971 arg1 unbinding 973:981 ligand unbinding 966:981 ligand unbinding 966:981 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 5 7 theme allosteric 732:741 arg1 site 743:746 an allosteric site 729:746 an allosteric site 729:746 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 5 8 theme extracellular 799:811 arg1 vestibule 813:821 the extracellular vestibule 795:821 the extracellular vestibule 795:821 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 3 9 theme X-ray 373:377 arg1 structures 396:405 X-ray crystallographic structures 373:405 X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine 373:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 4 10 theme serotonin 674:682 arg1 binding 684:690 serotonin binding 674:690 serotonin binding 674:690 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 3 11 theme crystallographic 379:394 arg1 structures 396:405 X-ray crystallographic structures 373:405 X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine 373:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 4 12 from SERT 522:525 arg1 conformation 546:557 an outward-open conformation 530:557 an outward-open conformation by lodging 530:568 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 1 13 theme serotonin 71:79 arg1 SERT 94:97 SERT 94:97 SERT 94:97 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 27049939 1 13 theme serotonin 71:79 arg1 transporter 81:91 The serotonin transporter 67:91 The serotonin transporter (SERT) 67:98 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 27049939 5 14 theme site 743:746 arg1 location 717:724 the location 713:724 the location of an allosteric site in the complex 713:761 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 7 15 theme antidepressant 1130:1143 arg1 action 1145:1150 antidepressant action 1130:1150 antidepressant action in SERT 1130:1158 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 7 16 from action 1145:1150 arg1 SERT 1155:1158 SERT 1155:1158 SERT 1155:1158 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 1 17 theme neurotransmitter 190:205 arg1 reuptake 178:185 the sodium- and chloride-dependent reuptake 143:185 the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons 143:230 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 27049939 0 18 theme X-ray 0:4 arg1 structures 6:15 X-ray structures 0:15 X-ray structures 0:15 X-ray structures and mechanism of the human serotonin transporter. 27049939 2 19 theme neurotransmitter 329:344 arg1 signalling 346:355 neurotransmitter signalling 329:355 neurotransmitter signalling 329:355 SERT is a target for antidepressant and psychostimulant drugs, which block reuptake and prolong neurotransmitter signalling. 27049939 7 20 from SERT 1155:1158 arg1 mechanism 1117:1125 the mechanism 1113:1125 the mechanism of antidepressant action in SERT 1113:1158 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 3 21 theme human 410:414 arg1 SERT 416:419 human SERT 410:419 human SERT 410:419 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 4 22 theme binding 585:591 arg1 site 593:596 the central binding site 573:596 the central binding site 573:596 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 1 23 theme presynaptic 212:222 arg1 neurons 224:230 presynaptic neurons 212:230 presynaptic neurons 212:230 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 27049939 3 24 theme SERT 416:419 arg1 structures 396:405 X-ray crystallographic structures 373:405 X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine 373:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 4 25 theme located 599:605 arg1 site 593:596 the central binding site 573:596 the central binding site 573:596 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 6 26 theme S 1050:1050 arg1 -citalopram 1052:1062 (S)-citalopram 1049:1062 (S)-citalopram 1049:1062 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 7 27 theme action 1145:1150 arg1 mechanism 1117:1125 the mechanism 1113:1125 the mechanism of antidepressant action in SERT 1113:1158 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 3 28 dep antidepressants 455:469 arg1 antidepressants 455:469 the antidepressants 451:469 the antidepressants (S)-citalopram or paroxetine 451:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 3 28 dep antidepressants 455:469 arg1 paroxetine 489:498 paroxetine 489:498 paroxetine 489:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 3 28 dep antidepressants 455:469 arg1 -citalopram 474:484 -citalopram 474:484 -citalopram 474:484 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 5 29 theme extracellular 843:855 arg1 loops 857:861 extracellular loops 843:861 extracellular loops 4 and 6 843:869 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 5 30 theme vestibule 813:821 arg1 periphery 782:790 the periphery 778:790 the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11 778:911 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 5 31 from location 717:724 arg1 complex 755:761 the complex 751:761 the complex 751:761 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 0 32 theme serotonin 44:52 arg1 transporter 54:64 the human serotonin transporter 34:64 the human serotonin transporter 34:64 X-ray structures and mechanism of the human serotonin transporter. 27049939 6 33 theme site 942:945 arg1 Occupancy 914:922 Occupancy 914:922 Occupancy of the allosteric site 914:945 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 0 34 theme human 38:42 arg1 transporter 54:64 the human serotonin transporter 34:64 the human serotonin transporter 34:64 X-ray structures and mechanism of the human serotonin transporter. 27049939 4 35 theme central 577:583 arg1 site 593:596 the central binding site 573:596 the central binding site 573:596 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 4 36 theme outward-open 533:544 arg1 conformation 546:557 an outward-open conformation 530:557 an outward-open conformation by lodging 530:568 Antidepressants lock SERT in an outward-open conformation by lodging in the central binding site, located between transmembrane helices 1, 3, 6, 8 and 10, directly blocking serotonin binding. 27049939 1 37 theme serotonergic 111:122 arg1 signalling 124:133 serotonergic signalling 111:133 serotonergic signalling 111:133 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 27049939 6 38 theme allosteric 1070:1079 arg1 explanation 1019:1029 an explanation 1016:1029 an explanation for the action of (S)-citalopram 1016:1062 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 6 38 theme allosteric 1070:1079 arg1 ligand 1081:1086 an allosteric ligand 1067:1086 an allosteric ligand 1067:1086 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 2 39 theme psychostimulant 273:287 arg1 drugs 289:293 antidepressant and psychostimulant drugs 254:293 antidepressant and psychostimulant drugs 254:293 SERT is a target for antidepressant and psychostimulant drugs, which block reuptake and prolong neurotransmitter signalling. 27049939 6 40 theme -citalopram 1052:1062 arg1 action 1039:1044 the action 1035:1044 the action of (S)-citalopram 1035:1062 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 5 41 theme transmembrane 875:887 arg1 helices 889:895 transmembrane helices 875:895 transmembrane helices 1, 6, 10 and 11 875:911 We further identify the location of an allosteric site in the complex as residing at the periphery of the extracellular vestibule, interposed between extracellular loops 4 and 6 and transmembrane helices 1, 6, 10 and 11. 27049939 7 42 theme future 1188:1193 arg1 design 1200:1205 future drug design 1188:1205 future drug design 1188:1205 These structures define the mechanism of antidepressant action in SERT, and provide blueprints for future drug design. 27049939 6 43 theme central 992:998 arg1 site 1000:1003 the central site 988:1003 the central site 988:1003 Occupancy of the allosteric site sterically hinders ligand unbinding from the central site, providing an explanation for the action of (S)-citalopram as an allosteric ligand. 27049939 0 44 theme transporter 54:64 arg1 mechanism 21:29 mechanism 21:29 mechanism of the human serotonin transporter 21:64 X-ray structures and mechanism of the human serotonin transporter. 27049939 0 44 theme transporter 54:64 arg1 structures 6:15 X-ray structures 0:15 X-ray structures 0:15 X-ray structures and mechanism of the human serotonin transporter. 27049939 3 45 from resolution 431:440 arg1 structures 396:405 X-ray crystallographic structures 373:405 X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine 373:498 Here we report X-ray crystallographic structures of human SERT at 3.15 Å resolution bound to the antidepressants (S)-citalopram or paroxetine. 27049939 2 46 theme antidepressant 254:267 arg1 drugs 289:293 antidepressant and psychostimulant drugs 254:293 antidepressant and psychostimulant drugs 254:293 SERT is a target for antidepressant and psychostimulant drugs, which block reuptake and prolong neurotransmitter signalling. 27049939 1 47 theme sodium- 147:153 arg1 reuptake 178:185 the sodium- and chloride-dependent reuptake 143:185 the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons 143:230 The serotonin transporter (SERT) terminates serotonergic signalling through the sodium- and chloride-dependent reuptake of neurotransmitter into presynaptic neurons. 21757827 5 0 from apparatus 804:812 arg1 O-glycosylated 776:789 O-glycosylated 776:789 O-glycosylated 776:789 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 2 1 theme human 294:298 arg1 Yif1p 300:304 human Yif1p 294:304 human Yif1p 294:304 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 4 2 theme predicted 528:536 arg1 weight 548:553 its predicted molecular weight 524:553 its predicted molecular weight 524:553 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 0 3 theme family 69:74 arg1 proteins 76:83 Yip domain family proteins 58:83 Yip domain family proteins 58:83 Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins. 21757827 5 4 theme mobility 832:839 arg1 form 841:844 a lower mobility form 824:844 a lower mobility form (46 kDa) 824:853 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 4 theme mobility 832:839 arg1 kDa 850:852 46 kDa 847:852 46 kDa 847:852 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 1 5 theme Golgi 206:210 arg1 transport 212:220 Golgi transport 206:220 Golgi transport 206:220 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 7 6 theme Golgi 1234:1238 arg1 structure 1240:1248 the Golgi structure 1230:1248 the Golgi structure 1230:1248 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 5 7 theme higher 877:882 arg1 form 893:896 a higher mobility form 875:896 a higher mobility form cleaved at its C-terminal luminal domain (36 kDa) 875:946 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 8 theme mobility 884:891 arg1 form 893:896 a higher mobility form 875:896 a higher mobility form cleaved at its C-terminal luminal domain (36 kDa) 875:946 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 4 9 with consistent 508:517 arg1 weight 548:553 its predicted molecular weight 524:553 its predicted molecular weight 524:553 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 4 10 theme molecular 538:546 arg1 weight 548:553 its predicted molecular weight 524:553 its predicted molecular weight 524:553 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 5 11 gly O-glycosylated 776:789 arg1 apparatus 804:812 the Golgi apparatus 794:812 the Golgi apparatus 794:812 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 11 gly O-glycosylated 776:789 arg1 YIPF3 702:706 YIPF3 702:706 YIPF3 702:706 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 11 gly O-glycosylated 776:789 arg1 form 753:756 a N-glycosylated form 736:756 a N-glycosylated form (40 kDa) 736:765 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 11 gly O-glycosylated 776:789 arg2 form 753:756 a N-glycosylated form 736:756 a N-glycosylated form (40 kDa) 736:765 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 11 gly O-glycosylated 776:789 arg2 YIPF3 702:706 YIPF3 702:706 YIPF3 702:706 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 1 12 theme yeast 143:147 arg1 Yip1p 149:153 yeast Yip1p 143:153 yeast Yip1p 143:153 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 5 13 theme N-glycosylated 738:751 arg1 YIPF3 702:706 YIPF3 702:706 YIPF3 702:706 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 13 theme N-glycosylated 738:751 arg1 form 753:756 a N-glycosylated form 736:756 a N-glycosylated form (40 kDa) 736:765 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 13 theme N-glycosylated 738:751 arg1 kDa 762:764 40 kDa 759:764 40 kDa 759:764 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 1 14 theme Yip1p 149:153 arg1 homologues 129:138 homologues 129:138 homologues 129:138 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 14 theme Yip1p 149:153 arg1 proteins 116:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 7 15 theme YIPF3 1104:1108 arg1 knockdown 1091:1099 The knockdown 1087:1099 The knockdown of YIPF3 or YIPF4 in HeLa cells 1087:1131 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 2 16 theme YIPF3 263:267 arg1 characterization 243:258 the characterization 239:258 the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively 239:328 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 5 17 theme Golgi 798:802 arg1 apparatus 804:812 the Golgi apparatus 794:812 the Golgi apparatus 794:812 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 7 18 theme apparatus 1168:1176 arg1 fragmentation 1141:1153 fragmentation 1141:1153 fragmentation of the Golgi apparatus 1141:1176 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 4 19 theme western 613:619 arg1 blotting 621:628 western blotting 613:628 western blotting 613:628 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 4 20 theme mobility 494:501 arg1 form 503:506 a single mobility form 485:506 a single mobility form consistent with its predicted molecular weight 485:553 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 4 20 theme mobility 494:501 arg1 YIPF4 463:467 YIPF4 463:467 YIPF4 463:467 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 1 21 theme Yip1 90:93 arg1 homologues 129:138 homologues 129:138 homologues 129:138 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 21 theme Yip1 90:93 arg1 proteins 116:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 0 22 theme YIPF3 20:24 arg1 Characterization 0:15 Characterization 0:15 Characterization of YIPF3 and YIPF4, cis-Golgi 0:45 Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins. 21757827 1 23 theme domain 95:100 arg1 homologues 129:138 homologues 129:138 homologues 129:138 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 23 theme domain 95:100 arg1 proteins 116:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 3 24 theme immuno-electron 354:368 arg1 microscopy 370:379 immuno-electron microscopy 354:379 immuno-electron microscopy 354:379 Immunofluorescence and immuno-electron microscopy showed that both YIPF3 and YIPF4 are clearly concentrated in the cis-Golgi. 21757827 4 25 theme consistent 508:517 arg1 form 503:506 a single mobility form 485:506 a single mobility form consistent with its predicted molecular weight 485:553 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 4 25 theme consistent 508:517 arg1 YIPF4 463:467 YIPF4 463:467 YIPF4 463:467 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 2 26 theme Yip1p 310:314 arg1 homologues 280:289 homologues 280:289 homologues of human Yif1p and Yip1p 280:314 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 2 26 theme Yip1p 310:314 arg1 YIPF3 263:267 YIPF3 263:267 YIPF3 263:267 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 7 27 theme Golgi 1162:1166 arg1 apparatus 1168:1176 the Golgi apparatus 1158:1176 the Golgi apparatus 1158:1176 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 0 28 theme YIPF4 30:34 arg1 Characterization 0:15 Characterization 0:15 Characterization of YIPF3 and YIPF4, cis-Golgi 0:45 Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins. 21757827 6 29 theme proper 1053:1058 arg1 localization 1060:1071 their proper localization 1047:1071 their proper localization 1047:1071 YIPF3 and YIPF4 form a complex in the Golgi apparatus, and this was suggested to be important for their proper localization and function. 21757827 4 30 theme YIPF3 590:594 arg1 forms 581:585 three different mobility forms 556:585 three different mobility forms of YIPF3 556:594 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 7 31 theme structure 1240:1248 arg1 maintenance 1215:1225 the maintenance 1211:1225 the maintenance of the Golgi structure 1211:1248 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 1 32 theme Yif1p 159:163 arg1 homologues 129:138 homologues 129:138 homologues 129:138 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 32 theme Yif1p 159:163 arg1 proteins 116:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 33 theme family 102:107 arg1 homologues 129:138 homologues 129:138 homologues 129:138 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 33 theme family 102:107 arg1 proteins 116:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 0 34 theme cis-Golgi 37:45 arg1 Characterization 0:15 Characterization 0:15 Characterization of YIPF3 and YIPF4, cis-Golgi 0:45 Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins. 21757827 4 35 theme single 487:492 arg1 form 503:506 a single mobility form 485:506 a single mobility form consistent with its predicted molecular weight 485:553 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 4 35 theme single 487:492 arg1 YIPF4 463:467 YIPF4 463:467 YIPF4 463:467 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 5 36 theme Biochemical 631:641 arg1 experiments 666:676 Biochemical and immunofluorescence experiments 631:676 experiments 666:676 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 7 37 from knockdown 1091:1099 arg1 cells 1127:1131 HeLa cells 1122:1131 HeLa cells 1122:1131 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 5 38 from O-glycosylated 776:789 arg1 apparatus 804:812 the Golgi apparatus 794:812 the Golgi apparatus 794:812 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 1 39 theme YIPF 110:113 arg1 homologues 129:138 homologues 129:138 homologues 129:138 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 1 39 theme YIPF 110:113 arg1 proteins 116:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins 86:123 The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. 21757827 5 40 theme immunofluorescence 647:664 arg1 experiments 666:676 Biochemical and immunofluorescence experiments 631:676 experiments 666:676 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 41 gly N-glycosylated 738:751 arg1 YIPF3 702:706 YIPF3 702:706 YIPF3 702:706 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 41 gly N-glycosylated 738:751 arg1 form 753:756 a N-glycosylated form 736:756 a N-glycosylated form (40 kDa) 736:765 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 41 gly N-glycosylated 738:751 arg1 kDa 762:764 40 kDa 759:764 40 kDa 759:764 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 42 theme lower 826:830 arg1 form 841:844 a lower mobility form 824:844 a lower mobility form (46 kDa) 824:853 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 42 theme lower 826:830 arg1 kDa 850:852 46 kDa 847:852 46 kDa 847:852 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 2 43 theme YIPF4 273:277 arg1 characterization 243:258 the characterization 239:258 the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively 239:328 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 0 44 theme domain 62:67 arg1 proteins 76:83 Yip domain family proteins 58:83 Yip domain family proteins 58:83 Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins. 21757827 6 45 theme Golgi 987:991 arg1 apparatus 993:1001 the Golgi apparatus 983:1001 the Golgi apparatus 983:1001 YIPF3 and YIPF4 form a complex in the Golgi apparatus, and this was suggested to be important for their proper localization and function. 21757827 5 46 theme C-terminal 913:922 arg1 domain 932:937 its C-terminal luminal domain 909:937 its C-terminal luminal domain (36 kDa) 909:946 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 46 theme C-terminal 913:922 arg1 kDa 943:945 36 kDa 940:945 36 kDa 940:945 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 7 47 from involvement 1196:1206 arg1 maintenance 1215:1225 the maintenance 1211:1225 the maintenance of the Golgi structure 1211:1248 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 7 48 theme HeLa 1122:1125 arg1 cells 1127:1131 HeLa cells 1122:1131 HeLa cells 1122:1131 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 4 49 theme mobility 572:579 arg1 forms 581:585 three different mobility forms 556:585 three different mobility forms of YIPF3 556:594 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 21757827 0 50 theme Yip 58:60 arg1 proteins 76:83 Yip domain family proteins 58:83 Yip domain family proteins 58:83 Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins. 21757827 5 51 theme luminal 924:930 arg1 domain 932:937 its C-terminal luminal domain 909:937 its C-terminal luminal domain (36 kDa) 909:946 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 5 51 theme luminal 924:930 arg1 kDa 943:945 36 kDa 940:945 36 kDa 940:945 Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). 21757827 2 52 theme Yif1p 300:304 arg1 homologues 280:289 homologues 280:289 homologues of human Yif1p and Yip1p 280:314 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 2 52 theme Yif1p 300:304 arg1 YIPF3 263:267 YIPF3 263:267 YIPF3 263:267 Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. 21757827 7 53 theme YIPF4 1113:1117 arg1 knockdown 1091:1099 The knockdown 1087:1099 The knockdown of YIPF3 or YIPF4 in HeLa cells 1087:1131 The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure. 21757827 4 54 theme different 562:570 arg1 forms 581:585 three different mobility forms 556:585 three different mobility forms of YIPF3 556:594 While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. 31125343 6 0 dep affected 807:814 arg1 causing 865:871 causing 865:871 causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del) 865:946 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 6 0 dep affected 807:814 arg1 leading 831:837 leading 831:837 leading to skipping of exon 5 831:859 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 7 1 theme typical 1053:1059 arg1 site 1083:1086 a typical type I copper binding site 1051:1086 a typical type I copper binding site in HEPHL1 1051:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 11 2 theme hair 1603:1606 arg1 phenotype 1608:1616 the hair phenotype 1599:1616 the hair phenotype in our patient 1599:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 8 3 theme ferroxidase 1212:1222 arg1 activity 1224:1231 this ferroxidase activity 1207:1231 this ferroxidase activity 1207:1231 We demonstrated that HEPHL1 has ferroxidase activity and that the patient's two mutations exhibited loss of this ferroxidase activity. 31125343 0 4 theme hair 76:79 arg1 phenotype 81:89 an abnormal hair phenotype 64:89 an abnormal hair phenotype 64:89 Biallelic HEPHL1 variants impair ferroxidase activity and cause an abnormal hair phenotype. 31125343 4 5 theme novel 506:510 arg1 Hephaestin 525:534 Hephaestin like 1 525:541 Hephaestin like 1 (HEPHL1) 525:550 Recently, a novel ferroxidase, Hephaestin like 1 (HEPHL1), also known as zyklopen, was identified. 31125343 4 5 theme novel 506:510 arg1 ferroxidase 512:522 a novel ferroxidase 504:522 a novel ferroxidase 504:522 Recently, a novel ferroxidase, Hephaestin like 1 (HEPHL1), also known as zyklopen, was identified. 31125343 6 6 theme missense 789:796 arg1 mutation 798:805 The maternal missense mutation 776:805 The maternal missense mutation 776:805 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 5 7 with child 610:614 arg1 mutations 643:651 compound heterozygous mutations 621:651 compound heterozygous mutations in HEPHL1 (NM_001098672) 621:676 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 11 8 with consistent 1583:1592 arg1 phenotype 1608:1616 the hair phenotype 1599:1616 the hair phenotype in our patient 1599:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 7 9 dep c.3176T>C 972:980 arg1 p.Met1059Thr 983:994 p.Met1059Thr 983:994 c.3176T>C; p.Met1059Thr 972:994 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 7 10 from site 1083:1086 arg1 HEPHL1 1091:1096 HEPHL1 1091:1096 HEPHL1 1091:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 12 11 theme HEPHL1 1693:1698 arg1 function 1681:1688 the function 1677:1688 the function of HEPHL1 1677:1698 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 9 12 theme copper-dependent 1361:1376 arg1 oxidase 1392:1398 lysyl oxidase 1386:1398 lysyl oxidase 1386:1398 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 9 12 theme copper-dependent 1361:1376 arg1 enzyme 1378:1383 the copper-dependent enzyme 1357:1383 the copper-dependent enzyme 1357:1383 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 6 13 theme exon 854:857 arg1 skipping 842:849 skipping 842:849 skipping of exon 5 842:859 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 5 14 theme compound 621:628 arg1 mutations 643:651 compound heterozygous mutations 621:651 compound heterozygous mutations in HEPHL1 (NM_001098672) 621:676 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 12 15 theme altered 1714:1720 arg1 activity 1734:1741 altered ferroxidase activity 1714:1741 altered ferroxidase activity in hair growth and hair disorders 1714:1775 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 12 16 from activity 1734:1741 arg1 growth 1751:1756 hair growth 1746:1756 hair growth 1746:1756 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 12 16 from activity 1734:1741 arg1 disorders 1767:1775 hair disorders 1762:1775 hair disorders 1762:1775 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 8 17 theme activity 1224:1231 arg1 loss 1199:1202 loss 1199:1202 loss of this ferroxidase activity 1199:1231 We demonstrated that HEPHL1 has ferroxidase activity and that the patient's two mutations exhibited loss of this ferroxidase activity. 31125343 5 18 theme abnormal 697:704 arg1 pili 712:715 pili torti and trichorrhexis nodosa 712:746 pili torti and trichorrhexis nodosa 712:746 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 5 18 theme abnormal 697:704 arg1 hair 706:709 abnormal hair 697:709 abnormal hair (pili torti and trichorrhexis nodosa) 697:747 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 7 19 from HEPHL1 1091:1096 arg1 methionine 1024:1033 a highly conserved methionine 1005:1033 a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1 1005:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 7 19 from HEPHL1 1091:1096 arg1 part 1043:1046 part 1043:1046 part of a typical type I copper binding site in HEPHL1 1043:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 9 20 theme enzyme 1378:1383 arg1 activity 1345:1352 reduced activity 1337:1352 reduced activity of the copper-dependent enzyme, lysyl oxidase 1337:1398 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 12 21 theme ferroxidase 1722:1732 arg1 activity 1734:1741 altered ferroxidase activity 1714:1741 altered ferroxidase activity in hair growth and hair disorders 1714:1775 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 7 22 theme binding 1075:1081 arg1 site 1083:1086 a typical type I copper binding site 1051:1086 a typical type I copper binding site in HEPHL1 1051:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 11 23 theme Hephl1 1515:1520 arg1 model 1537:1541 a Hephl1 knockout mouse model 1513:1541 a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient 1513:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 11 23 theme Hephl1 1515:1520 arg1 viable 1552:1557 viable 1552:1557 viable 1552:1557 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 7 24 dep mutation 962:969 arg1 c.3176T>C 972:980 c.3176T>C 972:980 c.3176T>C; p.Met1059Thr 972:994 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 1 25 theme critical 170:177 arg1 processes 190:198 critical biological processes 170:198 critical biological processes 170:198 Maintenance of the correct redox status of iron is functionally important for critical biological processes. 31125343 0 26 theme HEPHL1 10:15 arg1 variants 17:24 Biallelic HEPHL1 variants 0:24 Biallelic HEPHL1 variants 0:24 Biallelic HEPHL1 variants impair ferroxidase activity and cause an abnormal hair phenotype. 31125343 7 27 from part 1043:1046 arg1 HEPHL1 1091:1096 HEPHL1 1091:1096 HEPHL1 1091:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 11 28 theme knockout 1522:1529 arg1 model 1537:1541 a Hephl1 knockout mouse model 1513:1541 a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient 1513:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 11 28 theme knockout 1522:1529 arg1 viable 1552:1557 viable 1552:1557 viable 1552:1557 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 1 29 theme biological 179:188 arg1 processes 190:198 critical biological processes 170:198 critical biological processes 170:198 Maintenance of the correct redox status of iron is functionally important for critical biological processes. 31125343 0 30 theme Biallelic 0:8 arg1 variants 17:24 Biallelic HEPHL1 variants 0:24 Biallelic HEPHL1 variants 0:24 Biallelic HEPHL1 variants impair ferroxidase activity and cause an abnormal hair phenotype. 31125343 7 31 theme site 1083:1086 arg1 methionine 1024:1033 a highly conserved methionine 1005:1033 a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1 1005:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 7 31 theme site 1083:1086 arg1 part 1043:1046 part 1043:1046 part of a typical type I copper binding site in HEPHL1 1043:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 9 32 theme intracellular 1304:1316 arg1 iron 1318:1321 intracellular iron 1304:1321 intracellular iron 1304:1321 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 2 33 theme ferric 306:311 arg1 iron 313:316 ferric iron 306:316 ferric iron 306:316 Multicopper ferroxidases play an important role in oxidizing ferrous iron, released from the cells, into ferric iron, which is subsequently distributed by transferrin. 31125343 7 34 theme type 1061:1064 arg1 site 1083:1086 a typical type I copper binding site 1051:1086 a typical type I copper binding site in HEPHL1 1051:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 11 35 from phenotype 1608:1616 arg1 patient 1625:1631 our patient 1621:1631 our patient 1621:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 10 36 theme biallelic 1442:1450 arg1 variants 1452:1459 the patient's biallelic variants 1428:1459 the patient's biallelic variants 1428:1459 These results suggest that the patient's biallelic variants are loss-of-function mutations. 31125343 10 36 theme biallelic 1442:1450 arg1 mutations 1482:1490 loss-of-function mutations 1465:1490 loss-of-function mutations 1465:1490 These results suggest that the patient's biallelic variants are loss-of-function mutations. 31125343 12 37 theme function 1681:1688 arg1 understanding 1660:1672 our understanding 1656:1672 our understanding of the function of HEPHL1 1656:1698 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 3 38 theme well-characterized 373:390 arg1 ferroxidases 392:403 Two well-characterized ferroxidases 369:403 Two well-characterized ferroxidases 369:403 Two well-characterized ferroxidases, ceruloplasmin (CP) and hephaestin (HEPH) facilitate this reaction in different tissues. 31125343 3 38 theme well-characterized 373:390 arg1 hephaestin 429:438 hephaestin 429:438 hephaestin (HEPH) 429:445 Two well-characterized ferroxidases, ceruloplasmin (CP) and hephaestin (HEPH) facilitate this reaction in different tissues. 31125343 3 38 theme well-characterized 373:390 arg1 ceruloplasmin 406:418 ceruloplasmin 406:418 ceruloplasmin (CP) 406:423 Two well-characterized ferroxidases, ceruloplasmin (CP) and hephaestin (HEPH) facilitate this reaction in different tissues. 31125343 6 39 theme acids 906:910 arg1 deletion 885:892 an in-frame deletion 873:892 an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del) 873:946 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 7 40 theme conserved 1014:1022 arg1 methionine 1024:1033 a highly conserved methionine 1005:1033 a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1 1005:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 7 40 theme conserved 1014:1022 arg1 part 1043:1046 part 1043:1046 part of a typical type I copper binding site in HEPHL1 1043:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 5 41 theme trichorrhexis 727:739 arg1 pili 712:715 pili torti and trichorrhexis nodosa 712:746 pili torti and trichorrhexis nodosa 712:746 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 5 41 theme trichorrhexis 727:739 arg1 nodosa 741:746 trichorrhexis nodosa 727:746 trichorrhexis nodosa 727:746 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 6 42 theme amino 900:904 arg1 acids 906:910 85 amino acids 897:910 85 amino acids (c.809_1063del; p.Leu271_ala355del) 897:946 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 2 43 attach released 276:283 arg1 cells 294:298 the cells 290:298 the cells 290:298 Multicopper ferroxidases play an important role in oxidizing ferrous iron, released from the cells, into ferric iron, which is subsequently distributed by transferrin. 31125343 2 43 attach released 276:283 arg2 iron 270:273 ferrous iron 262:273 ferrous iron 262:273 Multicopper ferroxidases play an important role in oxidizing ferrous iron, released from the cells, into ferric iron, which is subsequently distributed by transferrin. 31125343 11 44 contain had 1563:1565 arg2 whiskers 1573:1580 curly whiskers 1567:1580 curly whiskers 1567:1580 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 11 44 contain had 1563:1565 arg1 model 1537:1541 a Hephl1 knockout mouse model 1513:1541 a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient 1513:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 11 44 contain had 1563:1565 arg1 viable 1552:1557 viable 1552:1557 viable 1552:1557 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 0 45 theme ferroxidase 33:43 arg1 activity 45:52 ferroxidase activity 33:52 ferroxidase activity 33:52 Biallelic HEPHL1 variants impair ferroxidase activity and cause an abnormal hair phenotype. 31125343 5 46 dep pili 712:715 arg1 pili 712:715 pili torti and trichorrhexis nodosa 712:746 pili torti and trichorrhexis nodosa 712:746 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 5 46 dep pili 712:715 arg1 torti 717:721 torti 717:721 torti 717:721 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 5 46 dep pili 712:715 arg1 nodosa 741:746 trichorrhexis nodosa 727:746 trichorrhexis nodosa 727:746 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 7 47 theme I 1066:1066 arg1 site 1083:1086 a typical type I copper binding site 1051:1086 a typical type I copper binding site in HEPHL1 1051:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 5 48 theme heterozygous 630:641 arg1 mutations 643:651 compound heterozygous mutations 621:651 compound heterozygous mutations in HEPHL1 (NM_001098672) 621:676 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 6 49 theme mRNA 816:819 arg1 splicing 821:828 mRNA splicing 816:828 mRNA splicing 816:828 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 2 50 theme important 234:242 arg1 role 244:247 an important role 231:247 an important role 231:247 Multicopper ferroxidases play an important role in oxidizing ferrous iron, released from the cells, into ferric iron, which is subsequently distributed by transferrin. 31125343 6 51 dep c.809_1063del 913:925 arg1 p.Leu271_ala355del 928:945 p.Leu271_ala355del 928:945 c.809_1063del; p.Leu271_ala355del 913:945 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 1 52 theme correct 111:117 arg1 status 125:130 the correct redox status 107:130 the correct redox status of iron 107:138 Maintenance of the correct redox status of iron is functionally important for critical biological processes. 31125343 7 53 theme copper 1068:1073 arg1 site 1083:1086 a typical type I copper binding site 1051:1086 a typical type I copper binding site in HEPHL1 1051:1096 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 4 54 theme like 536:539 arg1 Hephaestin 525:534 Hephaestin like 1 525:541 Hephaestin like 1 (HEPHL1) 525:550 Recently, a novel ferroxidase, Hephaestin like 1 (HEPHL1), also known as zyklopen, was identified. 31125343 4 54 theme like 536:539 arg1 ferroxidase 512:522 a novel ferroxidase 504:522 a novel ferroxidase 504:522 Recently, a novel ferroxidase, Hephaestin like 1 (HEPHL1), also known as zyklopen, was identified. 31125343 4 54 theme like 536:539 arg1 HEPHL1 544:549 HEPHL1 544:549 HEPHL1 544:549 Recently, a novel ferroxidase, Hephaestin like 1 (HEPHL1), also known as zyklopen, was identified. 31125343 10 55 theme loss-of-function 1465:1480 arg1 variants 1452:1459 the patient's biallelic variants 1428:1459 the patient's biallelic variants 1428:1459 These results suggest that the patient's biallelic variants are loss-of-function mutations. 31125343 10 55 theme loss-of-function 1465:1480 arg1 mutations 1482:1490 loss-of-function mutations 1465:1490 loss-of-function mutations 1465:1490 These results suggest that the patient's biallelic variants are loss-of-function mutations. 31125343 6 56 dep acids 906:910 arg1 c.809_1063del 913:925 c.809_1063del 913:925 c.809_1063del; p.Leu271_ala355del 913:945 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 11 57 theme mouse 1531:1535 arg1 model 1537:1541 a Hephl1 knockout mouse model 1513:1541 a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient 1513:1631 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 11 57 theme mouse 1531:1535 arg1 viable 1552:1557 viable 1552:1557 viable 1552:1557 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 1 58 theme redox 119:123 arg1 status 125:130 the correct redox status 107:130 the correct redox status of iron 107:138 Maintenance of the correct redox status of iron is functionally important for critical biological processes. 31125343 6 59 theme in-frame 876:883 arg1 deletion 885:892 an in-frame deletion 873:892 an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del) 873:946 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 7 60 theme paternal 953:960 arg1 mutation 962:969 The paternal mutation 949:969 The paternal mutation (c.3176T>C; p.Met1059Thr) 949:995 The paternal mutation (c.3176T>C; p.Met1059Thr) changed a highly conserved methionine that is part of a typical type I copper binding site in HEPHL1. 31125343 1 61 theme status 125:130 arg1 Maintenance 92:102 Maintenance 92:102 Maintenance of the correct redox status of iron 92:138 Maintenance of the correct redox status of iron is functionally important for critical biological processes. 31125343 12 62 theme hair 1762:1765 arg1 disorders 1767:1775 hair disorders 1762:1775 hair disorders 1762:1775 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 2 63 theme Multicopper 201:211 arg1 ferroxidases 213:224 Multicopper ferroxidases 201:224 Multicopper ferroxidases 201:224 Multicopper ferroxidases play an important role in oxidizing ferrous iron, released from the cells, into ferric iron, which is subsequently distributed by transferrin. 31125343 8 64 contain has 1127:1129 arg1 HEPHL1 1120:1125 HEPHL1 1120:1125 HEPHL1 1120:1125 We demonstrated that HEPHL1 has ferroxidase activity and that the patient's two mutations exhibited loss of this ferroxidase activity. 31125343 8 64 contain has 1127:1129 arg2 activity 1143:1150 ferroxidase activity 1131:1150 ferroxidase activity 1131:1150 We demonstrated that HEPHL1 has ferroxidase activity and that the patient's two mutations exhibited loss of this ferroxidase activity. 31125343 3 65 theme different 475:483 arg1 tissues 485:491 different tissues 475:491 different tissues 475:491 Two well-characterized ferroxidases, ceruloplasmin (CP) and hephaestin (HEPH) facilitate this reaction in different tissues. 31125343 9 66 with Consistent 1234:1243 arg1 findings 1256:1263 these findings 1250:1263 these findings 1250:1263 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 8 67 theme ferroxidase 1131:1141 arg1 activity 1143:1150 ferroxidase activity 1131:1150 ferroxidase activity 1131:1150 We demonstrated that HEPHL1 has ferroxidase activity and that the patient's two mutations exhibited loss of this ferroxidase activity. 31125343 0 68 theme abnormal 67:74 arg1 phenotype 81:89 an abnormal hair phenotype 64:89 an abnormal hair phenotype 64:89 Biallelic HEPHL1 variants impair ferroxidase activity and cause an abnormal hair phenotype. 31125343 6 69 theme maternal 780:787 arg1 mutation 798:805 The maternal missense mutation 776:805 The maternal missense mutation 776:805 The maternal missense mutation affected mRNA splicing, leading to skipping of exon 5 and causing an in-frame deletion of 85 amino acids (c.809_1063del; p.Leu271_ala355del). 31125343 2 70 theme ferrous 262:268 arg1 iron 270:273 ferrous iron 262:273 ferrous iron 262:273 Multicopper ferroxidases play an important role in oxidizing ferrous iron, released from the cells, into ferric iron, which is subsequently distributed by transferrin. 31125343 11 71 theme curly 1567:1571 arg1 whiskers 1573:1580 curly whiskers 1567:1580 curly whiskers 1567:1580 Hence, we generated a Hephl1 knockout mouse model that was viable and had curly whiskers, consistent with the hair phenotype in our patient. 31125343 1 72 theme iron 135:138 arg1 status 125:130 the correct redox status 107:130 the correct redox status of iron 107:138 Maintenance of the correct redox status of iron is functionally important for critical biological processes. 31125343 9 73 theme lysyl 1386:1390 arg1 oxidase 1392:1398 lysyl oxidase 1386:1398 lysyl oxidase 1386:1398 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 9 73 theme lysyl 1386:1390 arg1 enzyme 1378:1383 the copper-dependent enzyme 1357:1383 the copper-dependent enzyme 1357:1383 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 9 74 theme reduced 1337:1343 arg1 activity 1345:1352 reduced activity 1337:1352 reduced activity of the copper-dependent enzyme, lysyl oxidase 1337:1398 Consistent with these findings, the patient's fibroblasts accumulated intracellular iron and exhibited reduced activity of the copper-dependent enzyme, lysyl oxidase. 31125343 12 75 theme hair 1746:1749 arg1 growth 1751:1756 hair growth 1746:1756 hair growth 1746:1756 These results enhance our understanding of the function of HEPHL1 and implicate altered ferroxidase activity in hair growth and hair disorders. 31125343 5 76 theme cognitive 753:761 arg1 dysfunction 763:773 cognitive dysfunction 753:773 cognitive dysfunction 753:773 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 5 77 from mutations 643:651 arg1 NM_001098672 664:675 NM_001098672 664:675 NM_001098672 664:675 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 31125343 5 77 from mutations 643:651 arg1 HEPHL1 656:661 HEPHL1 656:661 HEPHL1 (NM_001098672) 656:676 Here we report a child with compound heterozygous mutations in HEPHL1 (NM_001098672) who presented with abnormal hair (pili torti and trichorrhexis nodosa) and cognitive dysfunction. 23695682 7 0 theme different 942:950 arg1 polypeptide 961:971 a different receptor polypeptide 940:971 a different receptor polypeptide 940:971 This change favors, and is sufficient to account for, the selection of a different receptor polypeptide to complete the cytokine-receptor complex. 23695682 1 1 theme different 144:152 arg1 IL-17A-F 177:184 IL-17A-F 177:184 IL-17A-F 177:184 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 1 1 theme different 144:152 arg1 cytokines 166:174 six different homodimeric cytokines 140:174 six different homodimeric cytokines (IL-17A-F) 140:185 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 0 2 theme receptor 65:72 arg1 A 74:74 IL-17 receptor A 59:74 IL-17 receptor A 59:74 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 6 3 from side 752:755 arg1 Binding 726:732 Binding 726:732 Binding to IL-17RA at one side of the IL-17A molecule 726:778 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 1 4 theme homodimeric 154:164 arg1 IL-17A-F 177:184 IL-17A-F 177:184 IL-17A-F 177:184 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 1 4 theme homodimeric 154:164 arg1 cytokines 166:174 six different homodimeric cytokines 140:174 six different homodimeric cytokines (IL-17A-F) 140:185 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 8 5 theme structural 1020:1029 arg1 results 1031:1037 The structural results 1016:1037 The structural results 1016:1037 The structural results are supported by biophysical studies with IL-17A variants produced by site-directed mutagenesis. 23695682 6 6 theme conformational 790:803 arg1 change 805:810 a conformational change 788:810 a conformational change 788:810 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 2 7 theme host 283:286 arg1 defenses 288:295 host defenses 283:295 host defenses 283:295 Their interactions with IL-17 receptors A-E (IL-17RA-E) mediate host defenses while also contributing to inflammatory and autoimmune responses. 23695682 4 8 theme family 584:589 arg1 members 569:575 other members 563:575 other members of its family to allow signaling of different IL-17 cytokines 563:637 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 7 9 theme receptor 952:959 arg1 polypeptide 961:971 a different receptor polypeptide 940:971 a different receptor polypeptide 940:971 This change favors, and is sufficient to account for, the selection of a different receptor polypeptide to complete the cytokine-receptor complex. 23695682 3 10 dep engage 401:406 arg1 both 381:384 both 381:384 both 381:384 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 3 11 contain containing 427:436 arg2 molecule 470:477 one molecule 466:477 one molecule of IL-17RC 466:488 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 3 11 contain containing 427:436 arg1 complex 419:425 a receptor complex 408:425 a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC 408:488 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 3 11 contain containing 427:436 arg2 molecule 442:449 one molecule 438:449 one molecule of IL-17RA 438:460 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 4 12 with pairs 552:556 arg1 members 569:575 other members 563:575 other members of its family to allow signaling of different IL-17 cytokines 563:637 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 2 13 dep receptors 249:257 arg1 IL-17RA-E 264:272 IL-17RA-E 264:272 IL-17RA-E 264:272 Their interactions with IL-17 receptors A-E (IL-17RA-E) mediate host defenses while also contributing to inflammatory and autoimmune responses. 23695682 2 13 dep receptors 249:257 arg1 A-E 259:261 A-E 259:261 IL-17 receptors A-E (IL-17RA-E) 243:273 Their interactions with IL-17 receptors A-E (IL-17RA-E) mediate host defenses while also contributing to inflammatory and autoimmune responses. 23695682 6 14 theme molecule 771:778 arg1 side 752:755 one side 748:755 one side of the IL-17A molecule 748:778 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 3 15 theme IL-17RA 454:460 arg1 molecule 470:477 one molecule 466:477 one molecule of IL-17RC 466:488 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 3 15 theme IL-17RA 454:460 arg1 molecule 442:449 one molecule 438:449 one molecule of IL-17RA 438:460 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 6 16 theme IL-17A 861:866 arg1 IL-17A 861:866 IL-17A 861:866 IL-17A 861:866 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 6 16 theme IL-17A 861:866 arg1 site 853:856 the second, symmetry-related receptor site 815:856 the second, symmetry-related receptor site of IL-17A 815:866 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 6 17 theme IL-17A 764:769 arg1 molecule 771:778 the IL-17A molecule 760:778 the IL-17A molecule 760:778 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 6 18 dep second 819:824 arg1 symmetry-related 827:842 symmetry-related 827:842 symmetry-related 827:842 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 5 19 with structures 663:672 arg1 IL-17RA 717:723 IL-17RA 717:723 IL-17RA 717:723 Here we report crystal structures of homodimeric IL-17A and its complex with IL-17RA. 23695682 0 20 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of interleukin 17A 0:36 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 0 21 theme interleukin 22:32 arg1 17A 34:36 interleukin 17A 22:36 interleukin 17A 22:36 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 1 22 theme constituent 81:91 arg1 polypeptides 93:104 The constituent polypeptides 77:104 The constituent polypeptides of the interleukin-17 family 77:133 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 4 23 theme IL-17 623:627 arg1 cytokines 629:637 different IL-17 cytokines 613:637 different IL-17 cytokines 613:637 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 7 24 theme polypeptide 961:971 arg1 selection 927:935 the selection 923:935 the selection of a different receptor polypeptide 923:971 This change favors, and is sufficient to account for, the selection of a different receptor polypeptide to complete the cytokine-receptor complex. 23695682 4 25 theme other 563:567 arg1 members 569:575 other members 563:575 other members of its family to allow signaling of different IL-17 cytokines 563:637 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 8 26 theme biophysical 1056:1066 arg1 studies 1068:1074 biophysical studies 1056:1074 biophysical studies with IL-17A variants produced by site-directed mutagenesis 1056:1133 The structural results are supported by biophysical studies with IL-17A variants produced by site-directed mutagenesis. 23695682 4 27 theme different 613:621 arg1 cytokines 629:637 different IL-17 cytokines 613:637 different IL-17 cytokines 613:637 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 2 28 with interactions 225:236 arg1 receptors 249:257 IL-17 receptors 243:257 IL-17 receptors A-E (IL-17RA-E) 243:273 Their interactions with IL-17 receptors A-E (IL-17RA-E) mediate host defenses while also contributing to inflammatory and autoimmune responses. 23695682 8 29 theme IL-17A 1081:1086 arg1 variants 1088:1095 IL-17A variants 1081:1095 IL-17A variants produced by site-directed mutagenesis 1081:1133 The structural results are supported by biophysical studies with IL-17A variants produced by site-directed mutagenesis. 23695682 1 30 theme heterodimeric 195:207 arg1 IL-17A/F 209:216 the heterodimeric IL-17A/F 191:216 the heterodimeric IL-17A/F 191:216 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 0 31 theme 17A 34:36 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of interleukin 17A 0:36 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 0 31 theme 17A 34:36 arg1 complex 46:52 its complex 42:52 its complex with IL-17 receptor A 42:74 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 6 32 theme receptor 844:851 arg1 IL-17A 861:866 IL-17A 861:866 IL-17A 861:866 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 6 32 theme receptor 844:851 arg1 site 853:856 the second, symmetry-related receptor site 815:856 the second, symmetry-related receptor site of IL-17A 815:866 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 4 33 theme cytokines 629:637 arg1 signaling 600:608 signaling 600:608 signaling of different IL-17 cytokines 600:637 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 7 34 theme cytokine-receptor 989:1005 arg1 complex 1007:1013 the cytokine-receptor complex 985:1013 the cytokine-receptor complex 985:1013 This change favors, and is sufficient to account for, the selection of a different receptor polypeptide to complete the cytokine-receptor complex. 23695682 2 35 theme autoimmune 341:350 arg1 responses 352:360 autoimmune responses 341:360 autoimmune responses 341:360 Their interactions with IL-17 receptors A-E (IL-17RA-E) mediate host defenses while also contributing to inflammatory and autoimmune responses. 23695682 3 36 theme receptor 410:417 arg1 complex 419:425 a receptor complex 408:425 a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC 408:488 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 0 37 with structures 8:17 arg1 A 74:74 IL-17 receptor A 59:74 IL-17 receptor A 59:74 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 4 38 theme shared 531:536 arg1 IL-17RA 507:513 IL-17RA 507:513 IL-17RA 507:513 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 4 38 theme shared 531:536 arg1 receptor 538:545 a shared receptor 529:545 a shared receptor 529:545 More generally, IL-17RA appears to be a shared receptor that pairs with other members of its family to allow signaling of different IL-17 cytokines. 23695682 5 39 theme homodimeric 677:687 arg1 IL-17A 689:694 homodimeric IL-17A 677:694 homodimeric IL-17A 677:694 Here we report crystal structures of homodimeric IL-17A and its complex with IL-17RA. 23695682 6 40 theme second 819:824 arg1 IL-17A 861:866 IL-17A 861:866 IL-17A 861:866 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 6 40 theme second 819:824 arg1 site 853:856 the second, symmetry-related receptor site 815:856 the second, symmetry-related receptor site of IL-17A 815:866 Binding to IL-17RA at one side of the IL-17A molecule induces a conformational change in the second, symmetry-related receptor site of IL-17A. 23695682 8 41 theme site-directed 1109:1121 arg1 mutagenesis 1123:1133 site-directed mutagenesis 1109:1133 site-directed mutagenesis 1109:1133 The structural results are supported by biophysical studies with IL-17A variants produced by site-directed mutagenesis. 23695682 1 42 theme interleukin-17 113:126 arg1 family 128:133 the interleukin-17 family 109:133 the interleukin-17 family 109:133 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 5 43 theme IL-17A 689:694 arg1 structures 663:672 crystal structures 655:672 crystal structures of homodimeric IL-17A 655:694 Here we report crystal structures of homodimeric IL-17A and its complex with IL-17RA. 23695682 5 43 theme IL-17A 689:694 arg1 complex 704:710 its complex 700:710 its complex with IL-17RA 700:723 Here we report crystal structures of homodimeric IL-17A and its complex with IL-17RA. 23695682 0 44 with complex 46:52 arg1 A 74:74 IL-17 receptor A 59:74 IL-17 receptor A 59:74 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 1 45 theme family 128:133 arg1 polypeptides 93:104 The constituent polypeptides 77:104 The constituent polypeptides of the interleukin-17 family 77:133 The constituent polypeptides of the interleukin-17 family form six different homodimeric cytokines (IL-17A-F) and the heterodimeric IL-17A/F. 23695682 8 46 with studies 1068:1074 arg1 variants 1088:1095 IL-17A variants 1081:1095 IL-17A variants produced by site-directed mutagenesis 1081:1133 The structural results are supported by biophysical studies with IL-17A variants produced by site-directed mutagenesis. 23695682 3 47 theme IL-17RC 482:488 arg1 molecule 470:477 one molecule 466:477 one molecule of IL-17RC 466:488 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 3 47 theme IL-17RC 482:488 arg1 molecule 442:449 one molecule 438:449 one molecule of IL-17RA 438:460 IL-17A and IL-17F both preferentially engage a receptor complex containing one molecule of IL-17RA and one molecule of IL-17RC. 23695682 0 48 theme IL-17 59:63 arg1 A 74:74 IL-17 receptor A 59:74 IL-17 receptor A 59:74 Crystal structures of interleukin 17A and its complex with IL-17 receptor A. 23695682 5 49 with complex 704:710 arg1 IL-17RA 717:723 IL-17RA 717:723 IL-17RA 717:723 Here we report crystal structures of homodimeric IL-17A and its complex with IL-17RA. 23695682 5 50 theme crystal 655:661 arg1 structures 663:672 crystal structures 655:672 crystal structures of homodimeric IL-17A 655:694 Here we report crystal structures of homodimeric IL-17A and its complex with IL-17RA. 23695682 2 51 theme IL-17 243:247 arg1 receptors 249:257 IL-17 receptors 243:257 IL-17 receptors A-E (IL-17RA-E) 243:273 Their interactions with IL-17 receptors A-E (IL-17RA-E) mediate host defenses while also contributing to inflammatory and autoimmune responses. 12525165 0 0 theme phosphate 73:81 arg1 ion 83:85 a phosphate ion 71:85 a phosphate ion 71:85 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 0 1 theme inhibitor 189:197 arg1 design 199:204 inhibitor design 189:204 inhibitor design 189:204 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 2 2 from intermediate 396:407 arg1 mechanism 426:434 the catalytic mechanism 412:434 the catalytic mechanism 412:434 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 1 3 theme phosphatase 259:269 arg1 structure 225:233 The X-ray crystal structure 207:233 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion 207:307 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 0 4 from structures 8:17 arg1 complex 58:64 complex 58:64 complex 58:64 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 5 5 with complex 856:862 arg1 acid 927:930 alpha-benzylaminobenzylphosphonic acid 893:930 alpha-benzylaminobenzylphosphonic acid 893:930 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 5 with complex 856:862 arg1 derivative 881:890 a phosphate derivative 869:890 a phosphate derivative 869:890 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 6 from structure 836:844 arg1 complex 856:862 complex 856:862 complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), 856:967 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 4 7 theme crystallization 635:649 arg1 matrix 651:656 the crystallization matrix 631:656 the crystallization matrix 631:656 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 2 8 from mechanism 426:434 arg1 snapshot 374:381 a snapshot 372:381 a snapshot of the final intermediate in the catalytic mechanism 372:434 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 1 9 theme A 336:336 arg1 resolution 338:347 2.4 A resolution 332:347 2.4 A resolution 332:347 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 6 10 theme active 1107:1112 arg1 site 1114:1117 the active site 1103:1117 the active site 1103:1117 This structure gives new insight into the determinants of binding hydrophobic ligands within the active site and allows us to explain PAP's preference for aromatic substrates. 12525165 2 11 theme proton 482:487 arg1 donor 489:493 a proton donor 480:493 a proton donor in catalysis 480:506 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 5 12 theme potent 935:940 arg1 acid 927:930 alpha-benzylaminobenzylphosphonic acid 893:930 alpha-benzylaminobenzylphosphonic acid 893:930 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 12 theme potent 935:940 arg1 nM 964:965 IC(50) = 4 nM 953:965 IC(50) = 4 nM 953:965 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 12 theme potent 935:940 arg1 inhibitor 942:950 a potent inhibitor 933:950 a potent inhibitor (IC(50) = 4 nM) 933:966 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 0 13 from phosphatase 43:53 arg1 complex 58:64 complex 58:64 complex 58:64 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 4 14 theme substrate 795:803 arg1 selectivity 805:815 macromolecular substrate selectivity 780:815 macromolecular substrate selectivity 780:815 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 2 15 theme intermediate 396:407 arg1 snapshot 374:381 a snapshot 372:381 a snapshot of the final intermediate in the catalytic mechanism 372:434 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 4 16 theme Bound 606:610 arg1 molecules 616:624 Bound PEG molecules 606:624 Bound PEG molecules from the crystallization matrix 606:656 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 0 17 theme alpha-benzylaminobenzylphosphonic 91:123 arg1 acid 125:128 alpha-benzylaminobenzylphosphonic acid 91:128 alpha-benzylaminobenzylphosphonic acid 91:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 4 18 theme macromolecular 780:793 arg1 selectivity 805:815 macromolecular substrate selectivity 780:815 macromolecular substrate selectivity 780:815 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 6 19 theme hydrophobic 1076:1086 arg1 ligands 1088:1094 hydrophobic ligands 1076:1094 hydrophobic ligands within the active site 1076:1117 This structure gives new insight into the determinants of binding hydrophobic ligands within the active site and allows us to explain PAP's preference for aromatic substrates. 12525165 0 20 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid 0:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 3 21 theme hydrogen 526:533 arg1 bonds 535:539 eight hydrogen bonds 520:539 eight hydrogen bonds 520:539 A total of eight hydrogen bonds serve to strongly bind the phosphate ion within the active site. 12525165 4 22 theme PEG 612:614 arg1 molecules 616:624 Bound PEG molecules 606:624 Bound PEG molecules from the crystallization matrix 606:656 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 0 23 theme human 22:26 arg1 phosphatase 43:53 human prostatic acid phosphatase 22:53 human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid 22:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 1 24 with complex 280:286 arg1 ion 305:307 a phosphate ion 293:307 a phosphate ion 293:307 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 5 25 theme alpha-benzylaminobenzylphosphonic 893:925 arg1 acid 927:930 alpha-benzylaminobenzylphosphonic acid 893:930 alpha-benzylaminobenzylphosphonic acid 893:930 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 25 theme alpha-benzylaminobenzylphosphonic 893:925 arg1 inhibitor 942:950 a potent inhibitor 933:950 a potent inhibitor (IC(50) = 4 nM) 933:966 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 25 theme alpha-benzylaminobenzylphosphonic 893:925 arg1 derivative 881:890 a phosphate derivative 869:890 a phosphate derivative 869:890 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 3 26 theme active 593:598 arg1 site 600:603 the active site 589:603 the active site 589:603 A total of eight hydrogen bonds serve to strongly bind the phosphate ion within the active site. 12525165 4 27 theme channel 695:701 arg1 identification 675:688 the identification 671:688 the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity 671:815 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 1 28 theme X-ray 211:215 arg1 structure 225:233 The X-ray crystal structure 207:233 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion 207:307 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 0 29 theme acid 38:41 arg1 phosphatase 43:53 human prostatic acid phosphatase 22:53 human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid 22:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 2 30 theme final 390:394 arg1 intermediate 396:407 the final intermediate 386:407 the final intermediate in the catalytic mechanism 386:434 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 5 31 theme = 960:960 arg1 inhibitor 942:950 a potent inhibitor 933:950 a potent inhibitor (IC(50) = 4 nM) 933:966 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 31 theme = 960:960 arg1 nM 964:965 IC(50) = 4 nM 953:965 IC(50) = 4 nM 953:965 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 1 32 theme crystal 217:223 arg1 structure 225:233 The X-ray crystal structure 207:233 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion 207:307 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 0 33 theme prostatic 28:36 arg1 phosphatase 43:53 human prostatic acid phosphatase 22:53 human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid 22:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 1 34 theme phosphate 295:303 arg1 ion 305:307 a phosphate ion 293:307 a phosphate ion 293:307 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 0 35 theme mechanistic 141:151 arg1 picture 153:159 the mechanistic picture 137:159 the mechanistic picture 137:159 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 2 36 theme catalytic 416:424 arg1 mechanism 426:434 the catalytic mechanism 412:434 the catalytic mechanism 412:434 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 0 37 from complex 58:64 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid 0:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 3 38 theme bonds 535:539 arg1 total 511:515 A total 509:515 A total of eight hydrogen bonds 509:539 A total of eight hydrogen bonds serve to strongly bind the phosphate ion within the active site. 12525165 1 39 from structure 225:233 arg1 complex 280:286 complex 280:286 complex with a phosphate ion 280:307 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 0 40 theme phosphatase 43:53 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid 0:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 6 41 theme aromatic 1165:1172 arg1 substrates 1174:1183 aromatic substrates 1165:1183 aromatic substrates 1165:1183 This structure gives new insight into the determinants of binding hydrophobic ligands within the active site and allows us to explain PAP's preference for aromatic substrates. 12525165 4 42 theme molecular 751:759 arg1 recognition 761:771 molecular recognition 751:771 molecular recognition 751:771 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 5 43 theme IC 953:954 arg1 inhibitor 942:950 a potent inhibitor 933:950 a potent inhibitor (IC(50) = 4 nM) 933:966 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 43 theme IC 953:954 arg1 nM 964:965 IC(50) = 4 nM 953:965 IC(50) = 4 nM 953:965 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 3 44 theme phosphate 568:576 arg1 ion 578:580 the phosphate ion 564:580 the phosphate ion 564:580 A total of eight hydrogen bonds serve to strongly bind the phosphate ion within the active site. 12525165 2 45 from snapshot 374:381 arg1 mechanism 426:434 the catalytic mechanism 412:434 the catalytic mechanism 412:434 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 5 46 theme phosphate 871:879 arg1 acid 927:930 alpha-benzylaminobenzylphosphonic acid 893:930 alpha-benzylaminobenzylphosphonic acid 893:930 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 5 46 theme phosphate 871:879 arg1 derivative 881:890 a phosphate derivative 869:890 a phosphate derivative 869:890 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 0 47 theme new 171:173 arg1 insights 175:182 new insights 171:182 new insights into inhibitor design 171:204 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 2 48 from donor 489:493 arg1 catalysis 498:506 catalysis 498:506 catalysis 498:506 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 6 49 theme new 1031:1033 arg1 insight 1035:1041 new insight 1031:1041 new insight into the determinants of binding hydrophobic ligands within the active site 1031:1117 This structure gives new insight into the determinants of binding hydrophobic ligands within the active site and allows us to explain PAP's preference for aromatic substrates. 12525165 1 50 theme human 238:242 arg1 PAP 272:274 PAP 272:274 PAP 272:274 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 1 50 theme human 238:242 arg1 phosphatase 259:269 human prostatic acid phosphatase 238:269 human prostatic acid phosphatase (PAP) 238:275 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 0 51 with phosphatase 43:53 arg1 ion 83:85 a phosphate ion 71:85 a phosphate ion 71:85 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 0 51 with phosphatase 43:53 arg1 acid 125:128 alpha-benzylaminobenzylphosphonic acid 91:128 alpha-benzylaminobenzylphosphonic acid 91:128 Crystal structures of human prostatic acid phosphatase in complex with a phosphate ion and alpha-benzylaminobenzylphosphonic acid update the mechanistic picture and offer new insights into inhibitor design. 12525165 1 52 theme prostatic 244:252 arg1 PAP 272:274 PAP 272:274 PAP 272:274 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 1 52 theme prostatic 244:252 arg1 phosphatase 259:269 human prostatic acid phosphatase 238:269 human prostatic acid phosphatase (PAP) 238:275 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 2 53 theme Asp 469:471 arg1 role 461:464 the role 457:464 the role of Asp 258 as a proton donor in catalysis 457:506 This structure offers a snapshot of the final intermediate in the catalytic mechanism and does not support the role of Asp 258 as a proton donor in catalysis. 12525165 5 54 theme PAP 849:851 arg1 structure 836:844 the structure 832:844 the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), 832:967 Additionally, the structure of PAP in complex with a phosphate derivative, alpha-benzylaminobenzylphosphonic acid, a potent inhibitor (IC(50) = 4 nM), has been determined to 2.9 A resolution. 12525165 4 55 from matrix 651:656 arg1 molecules 616:624 Bound PEG molecules 606:624 Bound PEG molecules from the crystallization matrix 606:656 Bound PEG molecules from the crystallization matrix have allowed the identification of a channel within the molecule that likely plays a role in molecular recognition and in macromolecular substrate selectivity. 12525165 1 56 theme acid 254:257 arg1 PAP 272:274 PAP 272:274 PAP 272:274 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 12525165 1 56 theme acid 254:257 arg1 phosphatase 259:269 human prostatic acid phosphatase 238:269 human prostatic acid phosphatase (PAP) 238:275 The X-ray crystal structure of human prostatic acid phosphatase (PAP) in complex with a phosphate ion has been determined at 2.4 A resolution. 21615908 0 0 theme fibin 84:88 arg1 expression 24:33 the expression 20:33 the expression 20:33 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 0 0 theme fibin 84:88 arg1 activity 45:52 promoter activity 36:52 promoter activity 36:52 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 0 0 theme fibin 84:88 arg1 architecture 68:79 molecular architecture 58:79 molecular architecture 58:79 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 6 1 theme disulfide 970:978 arg1 bond 980:983 intermolecular disulfide bond 955:983 intermolecular disulfide bond 955:983 The two conserved cysteines participate in intermolecular disulfide bond and multimer formation. 21615908 5 2 gly glycosylated 889:900 arg2 Asn30 905:909 Asn30 905:909 Asn30 905:909 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 2 gly glycosylated 889:900 arg1 protein 819:825 an evolutionarily conserved protein 791:825 an evolutionarily conserved protein 791:825 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 2 gly glycosylated 889:900 arg1 Fibin 782:786 Fibin 782:786 Fibin 782:786 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 3 3 theme embryonic 495:503 arg1 development 505:515 embryonic development 495:515 embryonic development 495:515 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 2 4 from relevance 306:314 arg1 humans 328:333 humans 328:333 humans 328:333 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 2 4 from relevance 306:314 arg1 mammals 345:351 other mammals 339:351 other mammals 339:351 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 10 5 theme associated 1434:1443 arg1 co-factor 1445:1453 an unknown covalently-linked or associated co-factor 1402:1453 an unknown covalently-linked or associated co-factor 1402:1453 However, proper secretion presumably requires an unknown covalently-linked or associated co-factor. 21615908 1 6 theme essential 163:171 arg1 Fibin 102:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 1 6 theme essential 163:171 arg1 molecule 154:161 a secreted signal molecule 136:161 a secreted signal molecule essential for pectoral fin bud initiation in zebrafish 136:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 8 7 theme CONCLUSION 1152:1161 arg1 Fibin 1163:1167 CONCLUSION Fibin 1152:1167 CONCLUSION Fibin 1152:1167 CONCLUSION Fibin is functionally relevant during embryogenesis and adult life. 21615908 5 8 dep protein 819:825 arg1 glycosylated 889:900 glycosylated 889:900 is glycosylated at Asn30 886:909 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 8 dep protein 819:825 arg1 carries 828:834 carries 828:834 carries a cleavable signal peptide (amino acids 1-18) 828:880 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 2 9 theme biological 295:304 arg1 relevance 306:314 biological relevance 295:314 biological relevance 295:314 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 10 10 link covalently-linked 1413:1429 arg1 co-factor 1445:1453 an unknown covalently-linked or associated co-factor 1402:1453 an unknown covalently-linked or associated co-factor 1402:1453 However, proper secretion presumably requires an unknown covalently-linked or associated co-factor. 21615908 10 11 theme covalently-linked 1413:1429 arg1 co-factor 1445:1453 an unknown covalently-linked or associated co-factor 1402:1453 an unknown covalently-linked or associated co-factor 1402:1453 However, proper secretion presumably requires an unknown covalently-linked or associated co-factor. 21615908 4 12 theme manganese 766:774 arg1 ions 776:779 manganese ions 766:779 manganese ions 766:779 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 6 13 theme multimer 989:996 arg1 formation 998:1006 multimer formation 989:1006 multimer formation 989:1006 The two conserved cysteines participate in intermolecular disulfide bond and multimer formation. 21615908 5 14 theme signal 848:853 arg1 peptide 855:861 a cleavable signal peptide 836:861 a cleavable signal peptide (amino acids 1-18) 836:880 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 14 theme signal 848:853 arg1 acids 870:874 amino acids 1-18 864:879 amino acids 1-18 864:879 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 3 15 theme other 421:425 arg1 tissues 453:459 many other embryonic and adult mouse tissues 416:459 many other embryonic and adult mouse tissues 416:459 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 5 16 theme amino 864:868 arg1 peptide 855:861 a cleavable signal peptide 836:861 a cleavable signal peptide (amino acids 1-18) 836:880 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 16 theme amino 864:868 arg1 acids 870:874 amino acids 1-18 864:879 amino acids 1-18 864:879 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 17 theme conserved 809:817 arg1 protein 819:825 an evolutionarily conserved protein 791:825 an evolutionarily conserved protein 791:825 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 17 theme conserved 809:817 arg1 Fibin 782:786 Fibin 782:786 Fibin 782:786 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 9 18 theme secretory 1337:1345 arg1 pathway 1347:1353 the secretory pathway 1333:1353 the secretory pathway 1333:1353 Its expression is regulated by a number of cellular signalling pathways and the protein is routed via the secretory pathway. 21615908 7 19 theme endoplasmic 1099:1109 arg1 reticulum 1111:1119 the endoplasmic reticulum 1095:1119 the endoplasmic reticulum 1095:1119 Although fibin displays all features of a secretory protein, it is mostly retained in the endoplasmic reticulum when heterologously expressed. 21615908 2 20 theme molecular 268:276 arg1 architecture 278:289 molecular architecture 268:289 molecular architecture 268:289 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 1 21 theme pectoral 177:184 arg1 initiation 194:203 pectoral fin bud initiation 177:203 pectoral fin bud initiation in zebrafish 177:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 8 22 theme adult 1219:1223 arg1 life 1225:1228 adult life 1219:1228 adult life 1219:1228 CONCLUSION Fibin is functionally relevant during embryogenesis and adult life. 21615908 6 23 theme intermolecular 955:968 arg1 bond 980:983 intermolecular disulfide bond 955:983 intermolecular disulfide bond 955:983 The two conserved cysteines participate in intermolecular disulfide bond and multimer formation. 21615908 4 24 theme fibin 650:654 arg1 expression 656:665 fibin expression 650:665 fibin expression 650:665 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 25 theme signalling 742:751 arg1 pathways 753:760 the protein kinase C signalling pathways 721:760 the protein kinase C signalling pathways 721:760 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 7 26 theme protein 1061:1067 arg1 features 1037:1044 all features 1033:1044 all features of a secretory protein 1033:1067 Although fibin displays all features of a secretory protein, it is mostly retained in the endoplasmic reticulum when heterologously expressed. 21615908 1 27 theme fin 186:188 arg1 initiation 194:203 pectoral fin bud initiation 177:203 pectoral fin bud initiation in zebrafish 177:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 4 28 theme 2.5-kbp 548:554 arg1 fragment 573:580 A 2.5-kbp genomic sequence fragment 546:580 A 2.5-kbp genomic sequence fragment upstream of the coding sequence 546:612 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 28 theme 2.5-kbp 548:554 arg1 sufficient 617:626 sufficient 617:626 sufficient 617:626 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 1 29 theme BACKGROUND 91:100 arg1 molecule 154:161 a secreted signal molecule 136:161 a secreted signal molecule essential for pectoral fin bud initiation in zebrafish 136:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 1 29 theme BACKGROUND 91:100 arg1 Fibin 102:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 1 30 theme bud 190:192 arg1 initiation 194:203 pectoral fin bud initiation 177:203 pectoral fin bud initiation in zebrafish 177:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 0 31 theme expression 24:33 arg1 Characterization 0:15 Characterization 0:15 Characterization of the expression, promoter activity and molecular architecture of fibin. 0:89 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 4 32 theme C 740:740 arg1 pathways 753:760 the protein kinase C signalling pathways 721:760 the protein kinase C signalling pathways 721:760 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 10 33 theme unknown 1405:1411 arg1 co-factor 1445:1453 an unknown covalently-linked or associated co-factor 1402:1453 an unknown covalently-linked or associated co-factor 1402:1453 However, proper secretion presumably requires an unknown covalently-linked or associated co-factor. 21615908 5 34 theme cleavable 838:846 arg1 peptide 855:861 a cleavable signal peptide 836:861 a cleavable signal peptide (amino acids 1-18) 836:880 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 5 34 theme cleavable 838:846 arg1 acids 870:874 amino acids 1-18 864:879 amino acids 1-18 864:879 Fibin is an evolutionarily conserved protein, carries a cleavable signal peptide (amino acids 1-18) and is glycosylated at Asn30. 21615908 2 35 theme fibin 319:323 arg1 architecture 278:289 molecular architecture 268:289 molecular architecture 268:289 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 2 35 theme fibin 319:323 arg1 relevance 306:314 biological relevance 295:314 biological relevance 295:314 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 4 36 theme sequence 564:571 arg1 fragment 573:580 A 2.5-kbp genomic sequence fragment 546:580 A 2.5-kbp genomic sequence fragment upstream of the coding sequence 546:612 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 36 theme sequence 564:571 arg1 sufficient 617:626 sufficient 617:626 sufficient 617:626 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 37 theme pathways 753:760 arg1 ions 776:779 manganese ions 766:779 manganese ions 766:779 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 37 theme pathways 753:760 arg1 activators 707:716 activators 707:716 activators of the protein kinase C signalling pathways 707:760 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 37 theme pathways 753:760 arg1 glucocorticoids 690:704 glucocorticoids 690:704 glucocorticoids 690:704 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 2 38 dep architecture 278:289 arg1 the 264:266 the 264:266 the 264:266 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 0 39 theme promoter 36:43 arg1 activity 45:52 promoter activity 36:52 promoter activity 36:52 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 4 40 theme genomic 556:562 arg1 fragment 573:580 A 2.5-kbp genomic sequence fragment 546:580 A 2.5-kbp genomic sequence fragment upstream of the coding sequence 546:612 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 40 theme genomic 556:562 arg1 sufficient 617:626 sufficient 617:626 sufficient 617:626 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 3 41 theme embryonic 427:435 arg1 tissues 453:459 many other embryonic and adult mouse tissues 416:459 many other embryonic and adult mouse tissues 416:459 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 3 42 theme mouse 447:451 arg1 tissues 453:459 many other embryonic and adult mouse tissues 416:459 many other embryonic and adult mouse tissues 416:459 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 3 43 theme many 416:419 arg1 tissues 453:459 many other embryonic and adult mouse tissues 416:459 many other embryonic and adult mouse tissues 416:459 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 10 44 theme proper 1365:1370 arg1 secretion 1372:1380 proper secretion 1365:1380 proper secretion presumably 1365:1391 However, proper secretion presumably requires an unknown covalently-linked or associated co-factor. 21615908 3 45 theme skeletal 396:403 arg1 muscle 405:410 skeletal muscle 396:410 skeletal muscle 396:410 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 7 46 theme secretory 1051:1059 arg1 protein 1061:1067 a secretory protein 1049:1067 a secretory protein 1049:1067 Although fibin displays all features of a secretory protein, it is mostly retained in the endoplasmic reticulum when heterologously expressed. 21615908 0 47 theme activity 45:52 arg1 Characterization 0:15 Characterization 0:15 Characterization of the expression, promoter activity and molecular architecture of fibin. 0:89 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 3 48 dep RESULTS 354:360 arg1 expressed 371:379 expressed 371:379 is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions 368:543 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 9 49 theme cellular 1274:1281 arg1 pathways 1294:1301 cellular signalling pathways 1274:1301 cellular signalling pathways 1274:1301 Its expression is regulated by a number of cellular signalling pathways and the protein is routed via the secretory pathway. 21615908 1 50 from initiation 194:203 arg1 zebrafish 208:216 zebrafish 208:216 zebrafish 208:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 2 51 theme other 339:343 arg1 mammals 345:351 other mammals 339:351 other mammals 339:351 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 2 52 from architecture 278:289 arg1 humans 328:333 humans 328:333 humans 328:333 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 2 52 from architecture 278:289 arg1 mammals 345:351 other mammals 339:351 other mammals 339:351 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 6 53 theme conserved 920:928 arg1 cysteines 930:938 The two conserved cysteines 912:938 The two conserved cysteines 912:938 The two conserved cysteines participate in intermolecular disulfide bond and multimer formation. 21615908 9 54 theme signalling 1283:1292 arg1 pathways 1294:1301 cellular signalling pathways 1274:1301 cellular signalling pathways 1274:1301 Its expression is regulated by a number of cellular signalling pathways and the protein is routed via the secretory pathway. 21615908 4 55 theme kinase 733:738 arg1 C 740:740 protein kinase C 725:740 the protein kinase C signalling pathways 721:760 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 56 theme sequence 605:612 arg1 upstream 582:589 upstream 582:589 upstream 582:589 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 3 57 theme adult 529:533 arg1 functions 535:543 adult functions 529:543 adult functions 529:543 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 9 58 theme pathways 1294:1301 arg1 number 1264:1269 a number 1262:1269 a number of cellular signalling pathways 1262:1301 Its expression is regulated by a number of cellular signalling pathways and the protein is routed via the secretory pathway. 21615908 4 59 theme protein 725:731 arg1 C 740:740 protein kinase C 725:740 the protein kinase C signalling pathways 721:760 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 4 60 theme coding 598:603 arg1 sequence 605:612 the coding sequence 594:612 the coding sequence 594:612 A 2.5-kbp genomic sequence fragment upstream of the coding sequence is sufficient to drive and regulate fibin expression through stimulation by glucocorticoids, activators of the protein kinase C signalling pathways and manganese ions. 21615908 0 61 theme architecture 68:79 arg1 Characterization 0:15 Characterization 0:15 Characterization of the expression, promoter activity and molecular architecture of fibin. 0:89 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 1 62 theme secreted 138:145 arg1 Fibin 102:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 1 62 theme secreted 138:145 arg1 molecule 154:161 a secreted signal molecule 136:161 a secreted signal molecule essential for pectoral fin bud initiation in zebrafish 136:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 3 63 theme adult 441:445 arg1 tissues 453:459 many other embryonic and adult mouse tissues 416:459 many other embryonic and adult mouse tissues 416:459 RESULTS Fibin is expressed in cerebellum, skeletal muscle and many other embryonic and adult mouse tissues suggesting not only a role during embryonic development but also in adult functions. 21615908 0 64 theme molecular 58:66 arg1 architecture 68:79 molecular architecture 58:79 molecular architecture 58:79 Characterization of the expression, promoter activity and molecular architecture of fibin. 21615908 2 65 theme little 239:244 arg1 information 246:256 little information 239:256 little information about the molecular architecture and biological relevance of fibin in humans and other mammals 239:351 Currently, there is little information about the molecular architecture and biological relevance of fibin in humans and other mammals. 21615908 1 66 theme signal 147:152 arg1 Fibin 102:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin 91:106 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 21615908 1 66 theme signal 147:152 arg1 molecule 154:161 a secreted signal molecule 136:161 a secreted signal molecule essential for pectoral fin bud initiation in zebrafish 136:216 BACKGROUND Fibin was initially discovered as a secreted signal molecule essential for pectoral fin bud initiation in zebrafish. 29343645 8 0 theme Proteolytic 994:1004 arg1 processing 1006:1015 Proteolytic processing 994:1015 Proteolytic processing 994:1015 Proteolytic processing allows movement of an amphipathic helix possibly involved in substrate access at membranes. 29343645 3 1 theme host 313:316 arg1 enzyme 318:323 a host enzyme 311:323 a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert 311:424 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 3 1 theme host 313:316 arg1 hydrolase 291:299 Acyloxyacyl hydrolase 279:299 Acyloxyacyl hydrolase (AOAH) 279:306 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 6 2 theme acyl 780:783 arg1 chain 785:789 a secondary acyl chain 768:789 a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site 768:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 4 3 theme Gram-negative 501:513 arg1 infection 515:523 Gram-negative infection 501:523 Gram-negative infection 501:523 This activity is critical for recovery from immune tolerance that follows Gram-negative infection. 29343645 8 4 theme amphipathic 1039:1049 arg1 helix 1051:1055 an amphipathic helix 1036:1055 an amphipathic helix possibly involved in substrate access at membranes 1036:1106 Proteolytic processing allows movement of an amphipathic helix possibly involved in substrate access at membranes. 29343645 5 5 theme crystal 600:606 arg1 structure 608:616 its crystal structure 596:616 its crystal structure 596:616 To understand the molecular mechanism of AOAH function, we determined its crystal structure and its complex with LPS. 29343645 3 6 theme acyloxyacyl-linked 349:366 arg1 acids 375:379 secondary (acyloxyacyl-linked) fatty acids 338:379 secondary (acyloxyacyl-linked) fatty acids 338:379 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 6 7 theme hydrophobic 700:710 arg1 pocket 712:717 a large hydrophobic pocket 692:717 a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site 692:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 7 8 with contacts 896:903 arg1 LPS 934:936 LPS 934:936 LPS 934:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 7 8 with contacts 896:903 arg1 groups 924:929 the phosphate groups 910:929 the phosphate groups of LPS 910:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 4 9 from tolerance 478:486 arg1 recovery 457:464 recovery 457:464 recovery from immune tolerance that follows Gram-negative infection 457:523 This activity is critical for recovery from immune tolerance that follows Gram-negative infection. 29343645 6 10 theme saposin 733:739 arg1 domains 755:761 the saposin and catalytic domains 729:761 domains 755:761 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 6 11 theme large 694:698 arg1 pocket 712:717 a large hydrophobic pocket 692:717 a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site 692:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 3 12 theme secondary 338:346 arg1 acids 375:379 secondary (acyloxyacyl-linked) fatty acids 338:379 secondary (acyloxyacyl-linked) fatty acids 338:379 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 2 13 theme Proper 142:147 arg1 recognition 149:159 Proper recognition 142:159 Proper recognition of LPS by pattern-recognition receptors 142:199 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 29343645 8 14 theme helix 1051:1055 arg1 movement 1024:1031 movement 1024:1031 movement of an amphipathic helix possibly involved in substrate access at membranes 1024:1106 Proteolytic processing allows movement of an amphipathic helix possibly involved in substrate access at membranes. 29343645 5 15 theme function 572:579 arg1 mechanism 554:562 the molecular mechanism 540:562 the molecular mechanism of AOAH function 540:579 To understand the molecular mechanism of AOAH function, we determined its crystal structure and its complex with LPS. 29343645 1 16 theme potent 75:80 arg1 endotoxin 92:100 a potent bacterial endotoxin 73:100 a potent bacterial endotoxin that triggers the innate immune system 73:139 LPS is a potent bacterial endotoxin that triggers the innate immune system. 29343645 1 16 theme potent 75:80 arg1 LPS 66:68 LPS 66:68 LPS 66:68 LPS is a potent bacterial endotoxin that triggers the innate immune system. 29343645 4 17 theme immune 471:476 arg1 tolerance 478:486 immune tolerance 471:486 immune tolerance that follows Gram-negative infection 471:523 This activity is critical for recovery from immune tolerance that follows Gram-negative infection. 29343645 6 18 theme secondary 770:778 arg1 chain 785:789 a secondary acyl chain 768:789 a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site 768:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 1 19 theme bacterial 82:90 arg1 endotoxin 92:100 a potent bacterial endotoxin 73:100 a potent bacterial endotoxin that triggers the innate immune system 73:139 LPS is a potent bacterial endotoxin that triggers the innate immune system. 29343645 1 19 theme bacterial 82:90 arg1 LPS 66:68 LPS 66:68 LPS 66:68 LPS is a potent bacterial endotoxin that triggers the innate immune system. 29343645 0 20 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the mammalian lipopolysaccharide 0:52 Crystal structure of the mammalian lipopolysaccharide detoxifier. 29343645 7 21 theme phosphate 914:922 arg1 LPS 934:936 LPS 934:936 LPS 934:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 7 21 theme phosphate 914:922 arg1 groups 924:929 the phosphate groups 910:929 the phosphate groups of LPS 910:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 6 22 theme active 848:853 arg1 site 855:858 the active site 844:858 the active site 844:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 6 23 from site 855:858 arg1 tunnel 834:839 a narrow lateral hydrophobic tunnel 805:839 a narrow lateral hydrophobic tunnel at the active site 805:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 3 24 theme Acyloxyacyl 279:289 arg1 enzyme 318:323 a host enzyme 311:323 a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert 311:424 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 3 24 theme Acyloxyacyl 279:289 arg1 AOAH 302:305 AOAH 302:305 AOAH 302:305 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 3 24 theme Acyloxyacyl 279:289 arg1 hydrolase 291:299 Acyloxyacyl hydrolase 279:299 Acyloxyacyl hydrolase (AOAH) 279:306 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 3 25 theme fatty 369:373 arg1 acids 375:379 secondary (acyloxyacyl-linked) fatty acids 338:379 secondary (acyloxyacyl-linked) fatty acids 338:379 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 8 26 theme substrate 1078:1086 arg1 access 1088:1093 substrate access 1078:1093 substrate access at membranes 1078:1106 Proteolytic processing allows movement of an amphipathic helix possibly involved in substrate access at membranes. 29343645 0 27 theme lipopolysaccharide 35:52 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the mammalian lipopolysaccharide 0:52 Crystal structure of the mammalian lipopolysaccharide detoxifier. 29343645 2 28 theme pattern-recognition 171:189 arg1 receptors 191:199 pattern-recognition receptors 171:199 pattern-recognition receptors 171:199 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 29343645 2 29 theme acyl 245:248 arg1 chains 250:255 typically six acyl chains 231:255 typically six acyl chains 231:255 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 29343645 6 30 theme lipid 660:664 arg1 moiety 666:671 The substrate's lipid moiety 644:671 The substrate's lipid moiety 644:671 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 0 31 theme mammalian 25:33 arg1 lipopolysaccharide 35:52 the mammalian lipopolysaccharide 21:52 the mammalian lipopolysaccharide 21:52 Crystal structure of the mammalian lipopolysaccharide detoxifier. 29343645 5 32 with structure 608:616 arg1 LPS 639:641 LPS 639:641 LPS 639:641 To understand the molecular mechanism of AOAH function, we determined its crystal structure and its complex with LPS. 29343645 5 33 theme molecular 544:552 arg1 mechanism 554:562 the molecular mechanism 540:562 the molecular mechanism of AOAH function 540:579 To understand the molecular mechanism of AOAH function, we determined its crystal structure and its complex with LPS. 29343645 2 34 theme LPS 164:166 arg1 recognition 149:159 Proper recognition 142:159 Proper recognition of LPS by pattern-recognition receptors 142:199 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 29343645 6 35 theme catalytic 745:753 arg1 domains 755:761 the saposin and catalytic domains 729:761 domains 755:761 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 6 36 theme hydrophobic 822:832 arg1 tunnel 834:839 a narrow lateral hydrophobic tunnel 805:839 a narrow lateral hydrophobic tunnel at the active site 805:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 1 37 theme innate 120:125 arg1 system 134:139 the innate immune system 116:139 the innate immune system 116:139 LPS is a potent bacterial endotoxin that triggers the innate immune system. 29343645 2 38 theme full 212:215 arg1 complement 217:226 a full complement 210:226 a full complement of typically six acyl chains 210:255 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 29343645 2 39 theme lipid 264:268 arg1 portion 270:276 the lipid portion 260:276 the lipid portion 260:276 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 29343645 5 40 theme AOAH 567:570 arg1 function 572:579 AOAH function 567:579 AOAH function 567:579 To understand the molecular mechanism of AOAH function, we determined its crystal structure and its complex with LPS. 29343645 1 41 theme immune 127:132 arg1 system 134:139 the innate immune system 116:139 the innate immune system 116:139 LPS is a potent bacterial endotoxin that triggers the innate immune system. 29343645 8 42 from membranes 1098:1106 arg1 access 1088:1093 substrate access 1078:1093 substrate access at membranes 1078:1106 Proteolytic processing allows movement of an amphipathic helix possibly involved in substrate access at membranes. 29343645 3 43 link acyloxyacyl-linked 349:366 arg1 acids 375:379 secondary (acyloxyacyl-linked) fatty acids 338:379 secondary (acyloxyacyl-linked) fatty acids 338:379 Acyloxyacyl hydrolase (AOAH) is a host enzyme that removes secondary (acyloxyacyl-linked) fatty acids from LPS, rendering it immunologically inert. 29343645 6 44 theme lateral 814:820 arg1 tunnel 834:839 a narrow lateral hydrophobic tunnel 805:839 a narrow lateral hydrophobic tunnel at the active site 805:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 7 45 theme oligosaccharide 969:983 arg1 portion 985:991 its oligosaccharide portion 965:991 its oligosaccharide portion 965:991 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 5 46 with complex 626:632 arg1 LPS 639:641 LPS 639:641 LPS 639:641 To understand the molecular mechanism of AOAH function, we determined its crystal structure and its complex with LPS. 29343645 7 47 theme dispensable 884:894 arg1 contacts 896:903 dispensable contacts 884:903 dispensable contacts with the phosphate groups of LPS 884:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 6 48 theme narrow 807:812 arg1 tunnel 834:839 a narrow lateral hydrophobic tunnel 805:839 a narrow lateral hydrophobic tunnel at the active site 805:858 The substrate's lipid moiety is accommodated in a large hydrophobic pocket formed by the saposin and catalytic domains with a secondary acyl chain inserted into a narrow lateral hydrophobic tunnel at the active site. 29343645 7 49 theme LPS 934:936 arg1 LPS 934:936 LPS 934:936 LPS 934:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 7 49 theme LPS 934:936 arg1 groups 924:929 the phosphate groups 910:929 the phosphate groups of LPS 910:936 The enzyme establishes dispensable contacts with the phosphate groups of LPS but does not interact with its oligosaccharide portion. 29343645 2 50 theme chains 250:255 arg1 complement 217:226 a full complement 210:226 a full complement of typically six acyl chains 210:255 Proper recognition of LPS by pattern-recognition receptors requires a full complement of typically six acyl chains in the lipid portion. 25385546 5 0 theme heparan 811:817 arg1 sulphate 819:826 heparan sulphate 811:826 heparan sulphate 811:826 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 1 1 theme neuronal 155:162 arg1 extension 164:172 neuronal extension 155:172 neuronal extension 155:172 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 5 2 theme co-culture 959:968 arg1 assays 970:975 neuronal co-culture assays 950:975 neuronal co-culture assays 950:975 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 2 3 theme synaptic 363:370 arg1 organization 372:383 synaptic organization 363:383 synaptic organization 363:383 Unknown mechanisms govern the shift in RPTPσ function, from outgrowth promotion to synaptic organization. 25385546 6 4 theme potentiating 1174:1185 arg1 interactions 1187:1198 RPTPσ flexibility potentiating interactions 1156:1198 RPTPσ flexibility potentiating interactions with additional ligands 1156:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 5 5 theme neuronal 950:957 arg1 assays 970:975 neuronal co-culture assays 950:975 neuronal co-culture assays 950:975 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 3 6 theme same 598:601 arg1 cell 603:606 same cell 598:606 same cell 598:606 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 3 7 theme cis 593:595 arg1 ligands 635:641 both cis (same cell) and trans (opposite cell) ligands 588:641 both cis (same cell) and trans (opposite cell) ligands 588:641 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 3 8 theme RPTPσ 540:544 arg1 region 560:565 the RPTPσ extracellular region 536:565 the RPTPσ extracellular region 536:565 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 6 9 theme additional 1205:1214 arg1 ligands 1216:1222 additional ligands 1205:1222 additional ligands 1205:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 4 10 theme Crystal 644:650 arg1 structures 652:661 Crystal structures 644:661 Crystal structures of RPTPσ 644:670 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 25385546 5 11 theme synaptic 922:929 arg1 differentiation 931:945 TrkC-dependent synaptic differentiation 907:945 TrkC-dependent synaptic differentiation in neuronal co-culture assays 907:975 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 3 12 theme scattering 463:472 arg1 analyses 474:481 crystallographic, electron microscopic and small-angle X-ray scattering analyses 402:481 crystallographic, electron microscopic and small-angle X-ray scattering analyses 402:481 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 3 13 with interaction 571:581 arg1 ligands 635:641 both cis (same cell) and trans (opposite cell) ligands 588:641 both cis (same cell) and trans (opposite cell) ligands 588:641 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 6 14 theme synapse 1250:1256 arg1 formation 1258:1266 excitatory synapse formation 1239:1266 excitatory synapse formation 1239:1266 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 0 15 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for extracellular cis and trans RPTPσ 0:53 Structural basis for extracellular cis and trans RPTPσ signal competition in synaptogenesis. 25385546 6 16 theme flexibility 1162:1172 arg1 interactions 1187:1198 RPTPσ flexibility potentiating interactions 1156:1198 RPTPσ flexibility potentiating interactions with additional ligands 1156:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 3 17 dep trans 613:617 arg1 cell 629:632 opposite cell 620:632 opposite cell 620:632 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 6 18 with interactions 1187:1198 arg1 ligands 1216:1222 additional ligands 1205:1222 additional ligands 1205:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 6 19 theme RPTPσ 1156:1160 arg1 interactions 1187:1198 RPTPσ flexibility potentiating interactions 1156:1198 RPTPσ flexibility potentiating interactions with additional ligands 1156:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 3 20 from flexibility 521:531 arg1 region 560:565 the RPTPσ extracellular region 536:565 the RPTPσ extracellular region 536:565 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 4 21 theme glycosaminoglycan-binding 766:790 arg1 site 792:795 the glycosaminoglycan-binding site 762:795 the glycosaminoglycan-binding site 762:795 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 25385546 4 22 theme postsynaptic 685:696 arg1 detail 710:715 its postsynaptic ligand TrkC detail 681:715 its postsynaptic ligand TrkC detail 681:715 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 25385546 3 23 theme crystallographic 402:417 arg1 analyses 474:481 crystallographic, electron microscopic and small-angle X-ray scattering analyses 402:481 crystallographic, electron microscopic and small-angle X-ray scattering analyses 402:481 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 6 24 theme proteoglycan 1052:1063 arg1 layer 1065:1069 the presynaptic proteoglycan layer 1036:1069 the presynaptic proteoglycan layer 1036:1069 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 1 25 theme Receptor 93:100 arg1 phosphatase 119:129 Receptor protein tyrosine phosphatase 93:129 Receptor protein tyrosine phosphatase sigma (RPTPσ) 93:143 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 6 26 theme consequent 1132:1141 arg1 reduction 1143:1151 the consequent reduction 1128:1151 the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands 1128:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 0 27 theme extracellular 21:33 arg1 RPTPσ 49:53 extracellular cis and trans RPTPσ 21:53 extracellular cis and trans RPTPσ 21:53 Structural basis for extracellular cis and trans RPTPσ signal competition in synaptogenesis. 25385546 6 28 theme presynaptic 1040:1050 arg1 layer 1065:1069 the presynaptic proteoglycan layer 1036:1069 the presynaptic proteoglycan layer 1036:1069 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 2 29 theme RPTPσ 319:323 arg1 function 325:332 RPTPσ function 319:332 RPTPσ function 319:332 Unknown mechanisms govern the shift in RPTPσ function, from outgrowth promotion to synaptic organization. 25385546 6 30 from reduction 1143:1151 arg1 interactions 1187:1198 RPTPσ flexibility potentiating interactions 1156:1198 RPTPσ flexibility potentiating interactions with additional ligands 1156:1222 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 1 31 theme presynaptic 188:198 arg1 nexus 200:204 a presynaptic nexus 186:204 a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis 186:277 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 1 31 theme presynaptic 188:198 arg1 sigma 131:135 Receptor protein tyrosine phosphatase sigma 93:135 Receptor protein tyrosine phosphatase sigma (RPTPσ) 93:143 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 5 32 theme RPTPσ 872:876 arg1 binding 878:884 RPTPσ binding 872:884 RPTPσ binding 872:884 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 1 33 theme tyrosine 110:117 arg1 phosphatase 119:129 Receptor protein tyrosine phosphatase 93:129 Receptor protein tyrosine phosphatase sigma (RPTPσ) 93:143 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 3 34 theme sufficient 497:506 arg1 flexibility 521:531 sufficient inter-domain flexibility 497:531 sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands 497:641 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 3 35 theme microscopic 429:439 arg1 analyses 474:481 crystallographic, electron microscopic and small-angle X-ray scattering analyses 402:481 crystallographic, electron microscopic and small-angle X-ray scattering analyses 402:481 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 4 36 theme RPTPσ 666:670 arg1 structures 652:661 Crystal structures 644:661 Crystal structures of RPTPσ 644:670 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 25385546 0 37 theme cis 35:37 arg1 RPTPσ 49:53 extracellular cis and trans RPTPσ 21:53 extracellular cis and trans RPTPσ 21:53 Structural basis for extracellular cis and trans RPTPσ signal competition in synaptogenesis. 25385546 5 38 theme heparin 832:838 arg1 oligomers 840:848 heparin oligomers 832:848 heparin oligomers 832:848 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 3 39 theme opposite 620:627 arg1 cell 629:632 opposite cell 620:632 opposite cell 620:632 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 1 40 theme multiple 210:217 arg1 interactions 244:255 multiple protein and proteoglycan interactions 210:255 multiple protein and proteoglycan interactions during synaptogenesis 210:277 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 6 41 theme ectodomain 1010:1019 arg1 emergence 1021:1029 transient RPTPσ ectodomain emergence 994:1029 transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer 994:1069 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 1 42 theme phosphatase 119:129 arg1 nexus 200:204 a presynaptic nexus 186:204 a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis 186:277 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 1 42 theme phosphatase 119:129 arg1 RPTPσ 138:142 RPTPσ 138:142 RPTPσ 138:142 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 1 42 theme phosphatase 119:129 arg1 sigma 131:135 Receptor protein tyrosine phosphatase sigma 93:135 Receptor protein tyrosine phosphatase sigma (RPTPσ) 93:143 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 0 43 theme trans 43:47 arg1 RPTPσ 49:53 extracellular cis and trans RPTPσ 21:53 extracellular cis and trans RPTPσ 21:53 Structural basis for extracellular cis and trans RPTPσ signal competition in synaptogenesis. 25385546 6 44 theme RPTPσ 1004:1008 arg1 emergence 1021:1029 transient RPTPσ ectodomain emergence 994:1029 transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer 994:1069 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 3 45 theme trans 613:617 arg1 ligands 635:641 both cis (same cell) and trans (opposite cell) ligands 588:641 both cis (same cell) and trans (opposite cell) ligands 588:641 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 3 46 theme extracellular 546:558 arg1 region 560:565 the RPTPσ extracellular region 536:565 the RPTPσ extracellular region 536:565 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 4 47 theme interaction 720:730 arg1 surface 732:738 an interaction surface 717:738 an interaction surface partially overlapping the glycosaminoglycan-binding site 717:795 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 25385546 6 48 theme transient 994:1002 arg1 emergence 1021:1029 transient RPTPσ ectodomain emergence 994:1029 transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer 994:1069 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 2 49 theme Unknown 280:286 arg1 mechanisms 288:297 Unknown mechanisms 280:297 Unknown mechanisms 280:297 Unknown mechanisms govern the shift in RPTPσ function, from outgrowth promotion to synaptic organization. 25385546 2 50 theme outgrowth 340:348 arg1 promotion 350:358 outgrowth promotion 340:358 outgrowth promotion to synaptic organization 340:383 Unknown mechanisms govern the shift in RPTPσ function, from outgrowth promotion to synaptic organization. 25385546 1 51 theme protein 102:108 arg1 phosphatase 119:129 Receptor protein tyrosine phosphatase 93:129 Receptor protein tyrosine phosphatase sigma (RPTPσ) 93:143 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 1 52 theme protein 219:225 arg1 interactions 244:255 multiple protein and proteoglycan interactions 210:255 multiple protein and proteoglycan interactions during synaptogenesis 210:277 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 6 53 from layer 1065:1069 arg1 emergence 1021:1029 transient RPTPσ ectodomain emergence 994:1029 transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer 994:1069 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 3 54 dep cis 593:595 arg1 cell 603:606 same cell 598:606 same cell 598:606 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 2 55 from shift 310:314 arg1 function 325:332 RPTPσ function 319:332 RPTPσ function 319:332 Unknown mechanisms govern the shift in RPTPσ function, from outgrowth promotion to synaptic organization. 25385546 3 56 theme inter-domain 508:519 arg1 flexibility 521:531 sufficient inter-domain flexibility 497:531 sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands 497:641 Here, we report crystallographic, electron microscopic and small-angle X-ray scattering analyses, which reveal sufficient inter-domain flexibility in the RPTPσ extracellular region for interaction with both cis (same cell) and trans (opposite cell) ligands. 25385546 6 57 theme trans-synaptic 1104:1117 arg1 complex 1119:1125 a trans-synaptic complex 1102:1125 a trans-synaptic complex 1102:1125 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 5 58 from differentiation 931:945 arg1 assays 970:975 neuronal co-culture assays 950:975 neuronal co-culture assays 950:975 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 4 59 theme TrkC 705:708 arg1 detail 710:715 its postsynaptic ligand TrkC detail 681:715 its postsynaptic ligand TrkC detail 681:715 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 25385546 5 60 theme TrkC-dependent 907:920 arg1 differentiation 931:945 TrkC-dependent synaptic differentiation 907:945 TrkC-dependent synaptic differentiation in neuronal co-culture assays 907:975 Accordingly, heparan sulphate and heparin oligomers compete with TrkC for RPTPσ binding in vitro and disrupt TrkC-dependent synaptic differentiation in neuronal co-culture assays. 25385546 1 61 theme proteoglycan 231:242 arg1 interactions 244:255 multiple protein and proteoglycan interactions 210:255 multiple protein and proteoglycan interactions during synaptogenesis 210:277 Receptor protein tyrosine phosphatase sigma (RPTPσ) regulates neuronal extension and acts as a presynaptic nexus for multiple protein and proteoglycan interactions during synaptogenesis. 25385546 6 62 theme excitatory 1239:1248 arg1 formation 1258:1266 excitatory synapse formation 1239:1266 excitatory synapse formation 1239:1266 We propose that transient RPTPσ ectodomain emergence from the presynaptic proteoglycan layer allows capture by TrkC to form a trans-synaptic complex, the consequent reduction in RPTPσ flexibility potentiating interactions with additional ligands to orchestrate excitatory synapse formation. 25385546 4 63 theme ligand 698:703 arg1 detail 710:715 its postsynaptic ligand TrkC detail 681:715 its postsynaptic ligand TrkC detail 681:715 Crystal structures of RPTPσ bound to its postsynaptic ligand TrkC detail an interaction surface partially overlapping the glycosaminoglycan-binding site. 16362042 3 0 theme human 294:298 arg1 complex 309:315 a minimal human Gas6/Axl complex 284:315 a minimal human Gas6/Axl complex 284:315 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 1 theme Gas6/Axl 300:307 arg1 complex 309:315 a minimal human Gas6/Axl complex 284:315 a minimal human Gas6/Axl complex 284:315 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 8 2 theme opposite 1194:1201 arg1 faces 1203:1207 opposite faces 1194:1207 opposite faces of the newly formed beta-sheet 1194:1238 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 3 3 theme Gas6 480:483 arg1 Gas6 480:483 Gas6 480:483 Gas6 480:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 3 theme Gas6 480:483 arg1 domain 470:475 the first laminin G-like domain 445:475 the first laminin G-like domain of Gas6 445:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 6 4 theme Axl 869:871 arg1 dimerisation 873:884 Gas6-mediated Axl dimerisation 855:884 Gas6-mediated Axl dimerisation 855:884 Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. 16362042 5 5 theme receptor 708:715 arg1 experiments 728:738 receptor activation experiments 708:738 receptor activation experiments 708:738 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 5 6 dep sites 795:799 arg1 both 757:760 both 757:760 both 757:760 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 4 7 theme Gas6/Axl 555:562 arg1 contacts 564:571 two distinct Gas6/Axl contacts 542:571 two distinct Gas6/Axl contacts of very different size 542:594 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 5 8 theme activation 717:726 arg1 experiments 728:738 receptor activation experiments 708:738 receptor activation experiments 708:738 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 3 9 attach crosslinked 430:440 arg3 stoichiometry 344:356 stoichiometry 344:356 stoichiometry 344:356 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 10 theme laminin 455:461 arg1 Gas6 480:483 Gas6 480:483 Gas6 480:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 10 theme laminin 455:461 arg1 domain 470:475 the first laminin G-like domain 445:475 the first laminin G-like domain of Gas6 445:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 2 11 theme Axl 141:143 arg1 signalling 145:154 Axl signalling 141:154 Axl signalling 141:154 Axl signalling plays important roles in cancer, spermatogenesis, immunity, and platelet function. 16362042 5 12 theme transmembrane 829:841 arg1 signalling 843:852 productive transmembrane signalling 818:852 productive transmembrane signalling 818:852 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 3 13 theme Axl 411:413 arg1 ectodomain 415:424 the Axl ectodomain 407:424 the Axl ectodomain 407:424 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 4 14 theme distinct 546:553 arg1 contacts 564:571 two distinct Gas6/Axl contacts 542:571 two distinct Gas6/Axl contacts of very different size 542:594 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 3 15 theme crystal 243:249 arg1 structure 251:259 The crystal structure 239:259 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex 239:315 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 8 16 from contact 1108:1114 arg1 Specificity 1083:1093 Specificity 1083:1093 Specificity at the major contact 1083:1114 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 1 17 theme Axl 75:77 arg1 family 79:84 the Axl family 71:84 the Axl family 71:84 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 3 18 theme ectodomain 415:424 arg1 ectodomain 415:424 the Axl ectodomain 407:424 the Axl ectodomain 407:424 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 18 theme ectodomain 415:424 arg1 domains 396:402 the two immunoglobulin-like domains 368:402 the two immunoglobulin-like domains of the Axl ectodomain 368:424 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 8 19 theme charged 1163:1169 arg1 residues 1182:1189 charged and apolar residues 1163:1189 charged and apolar residues 1163:1189 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 5 20 theme major 766:770 arg1 sites 795:799 the major and minor Gas6 binding sites 762:799 both the major and minor Gas6 binding sites 757:799 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 7 21 theme other 1036:1040 arg1 Mer 1078:1080 Mer 1078:1080 Mer 1078:1080 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 7 21 theme other 1036:1040 arg1 Sky/Tyro3 1064:1072 Sky/Tyro3 1064:1072 Sky/Tyro3 1064:1072 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 7 21 theme other 1036:1040 arg1 receptors 1053:1061 the other Axl family receptors 1032:1061 the other Axl family receptors 1032:1061 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 1 22 theme family 79:84 arg1 kinases 60:66 Receptor tyrosine kinases 42:66 Receptor tyrosine kinases of the Axl family 42:84 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 5 23 theme Structure-based 652:666 arg1 mutagenesis 668:678 Structure-based mutagenesis 652:678 Structure-based mutagenesis 652:678 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 0 24 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for Gas6-Axl signalling 0:39 Structural basis for Gas6-Axl signalling. 16362042 8 25 theme beta-sheet 1229:1238 arg1 faces 1203:1207 opposite faces 1194:1207 opposite faces of the newly formed beta-sheet 1194:1238 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 4 26 theme edge 633:636 arg1 beta-strands 638:649 edge beta-strands 633:649 edge beta-strands 633:649 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 0 27 theme Gas6-Axl 21:28 arg1 signalling 30:39 Gas6-Axl signalling 21:39 Gas6-Axl signalling 21:39 Structural basis for Gas6-Axl signalling. 16362042 5 28 theme protein 681:687 arg1 assays 697:702 protein binding assays 681:702 protein binding assays 681:702 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 3 29 theme Gas6/Gas6 512:520 arg1 contacts 522:529 Gas6/Gas6 contacts 512:529 Gas6/Gas6 contacts 512:529 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 5 30 theme productive 818:827 arg1 signalling 843:852 productive transmembrane signalling 818:852 productive transmembrane signalling 818:852 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 5 31 theme binding 689:695 arg1 assays 697:702 protein binding assays 681:702 protein binding assays 681:702 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 4 32 theme size 591:594 arg1 contacts 564:571 two distinct Gas6/Axl contacts 542:571 two distinct Gas6/Axl contacts of very different size 542:594 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 3 33 theme immunoglobulin-like 376:394 arg1 ectodomain 415:424 the Axl ectodomain 407:424 the Axl ectodomain 407:424 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 33 theme immunoglobulin-like 376:394 arg1 domains 396:402 the two immunoglobulin-like domains 368:402 the two immunoglobulin-like domains of the Axl ectodomain 368:424 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 7 34 theme Axl 1042:1044 arg1 Mer 1078:1080 Mer 1078:1080 Mer 1078:1080 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 7 34 theme Axl 1042:1044 arg1 Sky/Tyro3 1064:1072 Sky/Tyro3 1064:1072 Sky/Tyro3 1064:1072 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 7 34 theme Axl 1042:1044 arg1 receptors 1053:1061 the other Axl family receptors 1032:1061 the other Axl family receptors 1032:1061 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 2 35 theme platelet 220:227 arg1 function 229:236 platelet function 220:236 platelet function 220:236 Axl signalling plays important roles in cancer, spermatogenesis, immunity, and platelet function. 16362042 4 36 theme featuring 602:610 arg1 interactions 612:623 featuring interactions 602:623 featuring interactions between edge beta-strands 602:649 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 5 37 theme minor 776:780 arg1 sites 795:799 the major and minor Gas6 binding sites 762:799 both the major and minor Gas6 binding sites 757:799 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 6 38 theme Gas6/Axl 944:951 arg1 complex 953:959 a high-affinity 1:1 Gas6/Axl complex 924:959 a high-affinity 1:1 Gas6/Axl complex forming first 924:973 Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. 16362042 7 39 theme family 1046:1051 arg1 Mer 1078:1080 Mer 1078:1080 Mer 1078:1080 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 7 39 theme family 1046:1051 arg1 Sky/Tyro3 1064:1072 Sky/Tyro3 1064:1072 Sky/Tyro3 1064:1072 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 7 39 theme family 1046:1051 arg1 receptors 1053:1061 the other Axl family receptors 1032:1061 the other Axl family receptors 1032:1061 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 2 40 theme important 162:170 arg1 roles 172:176 important roles 162:176 important roles 162:176 Axl signalling plays important roles in cancer, spermatogenesis, immunity, and platelet function. 16362042 8 41 theme apolar 1175:1180 arg1 residues 1182:1189 charged and apolar residues 1163:1189 charged and apolar residues 1163:1189 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 5 42 theme Gas6 782:785 arg1 sites 795:799 the major and minor Gas6 binding sites 762:799 both the major and minor Gas6 binding sites 757:799 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 6 43 theme 1:1 940:942 arg1 complex 953:959 a high-affinity 1:1 Gas6/Axl complex 924:959 a high-affinity 1:1 Gas6/Axl complex forming first 924:973 Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. 16362042 8 44 theme residues 1182:1189 arg1 segregation 1148:1158 the segregation 1144:1158 the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet 1144:1238 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 6 45 theme Gas6-mediated 855:867 arg1 dimerisation 873:884 Gas6-mediated Axl dimerisation 855:884 Gas6-mediated Axl dimerisation 855:884 Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. 16362042 3 46 theme G-like 463:468 arg1 Gas6 480:483 Gas6 480:483 Gas6 480:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 46 theme G-like 463:468 arg1 domain 470:475 the first laminin G-like domain 445:475 the first laminin G-like domain of Gas6 445:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 5 47 theme binding 787:793 arg1 sites 795:799 the major and minor Gas6 binding sites 762:799 both the major and minor Gas6 binding sites 757:799 Structure-based mutagenesis, protein binding assays and receptor activation experiments demonstrate that both the major and minor Gas6 binding sites are required for productive transmembrane signalling. 16362042 6 48 theme high-affinity 926:938 arg1 complex 953:959 a high-affinity 1:1 Gas6/Axl complex 924:959 a high-affinity 1:1 Gas6/Axl complex forming first 924:973 Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. 16362042 3 49 theme minimal 286:292 arg1 complex 309:315 a minimal human Gas6/Axl complex 284:315 a minimal human Gas6/Axl complex 284:315 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 7 50 theme binding 996:1002 arg1 site 1004:1007 Only the minor Gas6 binding site 976:1007 Only the minor Gas6 binding site 976:1007 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 1 51 theme vitamin 107:113 arg1 Gas6 135:138 the vitamin K-dependent protein Gas6 103:138 the vitamin K-dependent protein Gas6 103:138 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 8 52 theme major 1102:1106 arg1 contact 1108:1114 the major contact 1098:1114 the major contact 1098:1114 Specificity at the major contact is suggested to result from the segregation of charged and apolar residues to opposite faces of the newly formed beta-sheet. 16362042 1 53 theme K-dependent 115:125 arg1 Gas6 135:138 the vitamin K-dependent protein Gas6 103:138 the vitamin K-dependent protein Gas6 103:138 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 3 54 from resolution 270:279 arg1 structure 251:259 The crystal structure 239:259 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex 239:315 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 55 theme direct 494:499 arg1 Axl/Axl 501:507 no direct Axl/Axl 491:507 no direct Axl/Axl 491:507 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 7 56 theme minor 985:989 arg1 site 1004:1007 Only the minor Gas6 binding site 976:1007 Only the minor Gas6 binding site 976:1007 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 1 57 theme protein 127:133 arg1 Gas6 135:138 the vitamin K-dependent protein Gas6 103:138 the vitamin K-dependent protein Gas6 103:138 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 3 58 theme first 449:453 arg1 Gas6 480:483 Gas6 480:483 Gas6 480:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 58 theme first 449:453 arg1 domain 470:475 the first laminin G-like domain 445:475 the first laminin G-like domain of Gas6 445:483 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 3 59 theme complex 309:315 arg1 resolution 270:279 3.3 A resolution 264:279 3.3 A resolution of a minimal human Gas6/Axl complex 264:315 The crystal structure at 3.3 A resolution of a minimal human Gas6/Axl complex reveals an assembly of 2:2 stoichiometry, in which the two immunoglobulin-like domains of the Axl ectodomain are crosslinked by the first laminin G-like domain of Gas6, with no direct Axl/Axl or Gas6/Gas6 contacts. 16362042 4 60 dep contacts 564:571 arg1 both 597:600 both 597:600 both 597:600 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 1 61 theme Receptor 42:49 arg1 kinases 60:66 Receptor tyrosine kinases 42:66 Receptor tyrosine kinases of the Axl family 42:84 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 7 62 theme Gas6 991:994 arg1 site 1004:1007 Only the minor Gas6 binding site 976:1007 Only the minor Gas6 binding site 976:1007 Only the minor Gas6 binding site is highly conserved in the other Axl family receptors, Sky/Tyro3 and Mer. 16362042 4 63 theme different 581:589 arg1 size 591:594 very different size 576:594 very different size 576:594 There are two distinct Gas6/Axl contacts of very different size, both featuring interactions between edge beta-strands. 16362042 1 64 theme tyrosine 51:58 arg1 kinases 60:66 Receptor tyrosine kinases 42:66 Receptor tyrosine kinases of the Axl family 42:84 Receptor tyrosine kinases of the Axl family are activated by the vitamin K-dependent protein Gas6. 16362042 6 65 with likely 889:894 arg1 complex 953:959 a high-affinity 1:1 Gas6/Axl complex 924:959 a high-affinity 1:1 Gas6/Axl complex forming first 924:973 Gas6-mediated Axl dimerisation is likely to occur in two steps, with a high-affinity 1:1 Gas6/Axl complex forming first. 15738388 10 0 dep segments 1262:1269 arg1 shaping 1271:1277 shaping 1271:1277 shaping 1271:1277 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 7 1 theme inhibition 966:975 arg1 constant 977:984 a nanomolar inhibition constant 954:984 a nanomolar inhibition constant 954:984 The complex occludes a large contact surface but makes rather few contacts, despite a nanomolar inhibition constant. 15738388 5 2 theme connecting 759:768 arg1 segment 770:776 a connecting segment 757:776 a connecting segment encompassing a third alpha-helix 757:809 These subdomains are packed against each other through the helices and linked by a connecting segment encompassing a third alpha-helix. 15738388 2 3 with treatment 367:375 arg1 inhibitors 402:411 histone deacetylase inhibitors 382:411 histone deacetylase inhibitors 382:411 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 13 4 theme tissue 1695:1700 arg1 inhibitors 1702:1711 tissue inhibitors 1695:1711 tissue inhibitors of metalloproteases 1695:1731 Because they are ubiquitous, latexins could represent for vertebrate A/B MCPs the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases. 15738388 10 5 from differences 1238:1248 arg1 segments 1262:1269 the loop segments 1253:1269 the loop segments shaping the border of the funnel-like access to the protease active site 1253:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 10 6 theme Major 1232:1236 arg1 differences 1238:1248 Major differences 1232:1248 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site 1232:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 4 7 theme equivalent 564:573 arg1 reminiscent 587:597 reminiscent 587:597 reminiscent of cystatins 587:610 Human latexin consists of two topologically equivalent subdomains, reminiscent of cystatins, consisting of an alpha-helix enveloped by a curved beta-sheet. 15738388 4 7 theme equivalent 564:573 arg1 subdomains 575:584 two topologically equivalent subdomains 546:584 two topologically equivalent subdomains 546:584 Human latexin consists of two topologically equivalent subdomains, reminiscent of cystatins, consisting of an alpha-helix enveloped by a curved beta-sheet. 15738388 8 8 theme latexin 1036:1042 arg1 flexibility 1021:1031 the flexibility 1017:1031 the flexibility of latexin 1017:1042 This low specificity explains the flexibility of latexin in inhibiting all vertebrate A/B MCPs tested, even across species barriers. 15738388 3 9 theme characteristic 478:491 arg1 fold 514:517 the characteristic alpha/beta-hydrolase fold 474:517 the characteristic alpha/beta-hydrolase fold 474:517 hCPA4 is a member of the A/B subfamily of MCPs and displays the characteristic alpha/beta-hydrolase fold. 15738388 2 10 theme human 288:292 arg1 CPA4 294:297 human CPA4 288:297 human CPA4 (hCPA4) 288:305 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 2 10 theme human 288:292 arg1 hCPA4 300:304 hCPA4 300:304 hCPA4 300:304 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 11 11 theme ascribable 1401:1410 arg1 sequences 1391:1399 Several sequences 1383:1399 Several sequences ascribable to diverse tissues and organs 1383:1440 Several sequences ascribable to diverse tissues and organs have been identified in vertebrate genomes as being highly similar to latexin. 15738388 3 12 theme MCPs 456:459 arg1 subfamily 443:451 the A/B subfamily 435:451 the A/B subfamily of MCPs 435:459 hCPA4 is a member of the A/B subfamily of MCPs and displays the characteristic alpha/beta-hydrolase fold. 15738388 13 13 theme A/B 1666:1668 arg1 MCPs 1670:1673 vertebrate A/B MCPs 1655:1673 vertebrate A/B MCPs 1655:1673 Because they are ubiquitous, latexins could represent for vertebrate A/B MCPs the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases. 15738388 3 14 theme A/B 439:441 arg1 subfamily 443:451 the A/B subfamily 435:451 the A/B subfamily of MCPs 435:459 hCPA4 is a member of the A/B subfamily of MCPs and displays the characteristic alpha/beta-hydrolase fold. 15738388 11 15 theme Several 1383:1389 arg1 sequences 1391:1399 Several sequences 1383:1399 Several sequences ascribable to diverse tissues and organs 1383:1440 Several sequences ascribable to diverse tissues and organs have been identified in vertebrate genomes as being highly similar to latexin. 15738388 13 16 theme matrix 1737:1742 arg1 metalloproteinases 1744:1761 matrix metalloproteinases 1737:1761 matrix metalloproteinases 1737:1761 Because they are ubiquitous, latexins could represent for vertebrate A/B MCPs the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases. 15738388 8 17 theme low 992:994 arg1 specificity 996:1006 This low specificity 987:1006 This low specificity 987:1006 This low specificity explains the flexibility of latexin in inhibiting all vertebrate A/B MCPs tested, even across species barriers. 15738388 2 18 theme carboxypeptidase 251:266 arg1 inhibitor 268:276 alias endogenous carboxypeptidase inhibitor 234:276 alias endogenous carboxypeptidase inhibitor 234:276 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 2 18 theme carboxypeptidase 251:266 arg1 Latexin 225:231 Latexin 225:231 Latexin 225:231 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 10 19 theme loop 1257:1260 arg1 segments 1262:1269 the loop segments 1253:1269 the loop segments shaping the border of the funnel-like access to the protease active site 1253:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 10 20 theme protease 1323:1330 arg1 site 1339:1342 the protease active site 1319:1342 the protease active site 1319:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 8 21 theme vertebrate 1062:1071 arg1 MCPs 1077:1080 all vertebrate A/B MCPs 1058:1080 all vertebrate A/B MCPs tested 1058:1087 This low specificity explains the flexibility of latexin in inhibiting all vertebrate A/B MCPs tested, even across species barriers. 15738388 7 22 theme few 932:934 arg1 contacts 936:943 rather few contacts 925:943 rather few contacts 925:943 The complex occludes a large contact surface but makes rather few contacts, despite a nanomolar inhibition constant. 15738388 11 23 theme diverse 1415:1421 arg1 tissues 1423:1429 tissues 1423:1429 tissues 1423:1429 Several sequences ascribable to diverse tissues and organs have been identified in vertebrate genomes as being highly similar to latexin. 15738388 9 24 theme invertebrate 1191:1202 arg1 MCPs 1208:1211 MCPs and invertebrate A/B MCPs 1182:1211 MCPs and invertebrate A/B MCPs 1182:1211 In contrast, modeling studies reveal why the N/E subfamily of MCPs and invertebrate A/B MCPs are not inhibited. 15738388 2 25 theme cancer 348:353 arg1 cells 355:359 prostate cancer cells 339:359 prostate cancer cells 339:359 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 0 26 theme carboxypeptidase 19:34 arg1 A4 36:37 human carboxypeptidase A4 13:37 human carboxypeptidase A4 13:37 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 12 27 theme inhibitors 1585:1594 arg1 family 1565:1570 the latexin family 1553:1570 the latexin family of potential inhibitors 1553:1594 They are proposed to constitute the latexin family of potential inhibitors. 15738388 9 28 theme A/B 1204:1206 arg1 MCPs 1208:1211 MCPs and invertebrate A/B MCPs 1182:1211 MCPs and invertebrate A/B MCPs 1182:1211 In contrast, modeling studies reveal why the N/E subfamily of MCPs and invertebrate A/B MCPs are not inhibited. 15738388 9 29 theme N/E 1165:1167 arg1 subfamily 1169:1177 the N/E subfamily 1161:1177 the N/E subfamily of MCPs and invertebrate A/B MCPs 1161:1211 In contrast, modeling studies reveal why the N/E subfamily of MCPs and invertebrate A/B MCPs are not inhibited. 15738388 1 30 theme 25-kDa 181:186 arg1 protein 188:194 a 25-kDa protein 179:194 a 25-kDa protein discovered in the rat brain 179:222 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 1 30 theme 25-kDa 181:186 arg1 latexin 170:176 latexin 170:176 latexin 170:176 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 0 31 theme human 13:17 arg1 A4 36:37 human carboxypeptidase A4 13:37 human carboxypeptidase A4 13:37 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 8 32 theme A/B 1073:1075 arg1 MCPs 1077:1080 all vertebrate A/B MCPs 1058:1080 all vertebrate A/B MCPs tested 1058:1087 This low specificity explains the flexibility of latexin in inhibiting all vertebrate A/B MCPs tested, even across species barriers. 15738388 6 33 theme subdomains 858:867 arg1 interface 839:847 the interface 835:847 the interface of these subdomains 835:867 The enzyme is bound at the interface of these subdomains. 15738388 1 34 theme endogenous 96:105 arg1 inhibitor 115:123 The only endogenous protein inhibitor 87:123 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) 87:165 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 1 34 theme endogenous 96:105 arg1 latexin 170:176 latexin 170:176 latexin 170:176 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 2 35 theme deacetylase 390:400 arg1 inhibitors 402:411 histone deacetylase inhibitors 382:411 histone deacetylase inhibitors 382:411 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 13 36 theme metalloproteases 1716:1731 arg1 inhibitors 1702:1711 tissue inhibitors 1695:1711 tissue inhibitors of metalloproteases 1695:1731 Because they are ubiquitous, latexins could represent for vertebrate A/B MCPs the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases. 15738388 11 37 theme vertebrate 1466:1475 arg1 genomes 1477:1483 vertebrate genomes 1466:1483 vertebrate genomes 1466:1483 Several sequences ascribable to diverse tissues and organs have been identified in vertebrate genomes as being highly similar to latexin. 15738388 2 38 theme histone 382:388 arg1 inhibitors 402:411 histone deacetylase inhibitors 382:411 histone deacetylase inhibitors 382:411 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 0 39 theme A4 36:37 arg1 Structure 0:8 Structure 0:8 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 0:85 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 13 40 theme vertebrate 1655:1664 arg1 MCPs 1670:1673 vertebrate A/B MCPs 1655:1673 vertebrate A/B MCPs 1655:1673 Because they are ubiquitous, latexins could represent for vertebrate A/B MCPs the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases. 15738388 0 41 theme endogenous 48:57 arg1 latexin 78:84 latexin 78:84 latexin 78:84 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 0 41 theme endogenous 48:57 arg1 inhibitor 67:75 its endogenous protein inhibitor 44:75 its endogenous protein inhibitor 44:75 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 12 42 theme latexin 1557:1563 arg1 family 1565:1570 the latexin family 1553:1570 the latexin family of potential inhibitors 1553:1594 They are proposed to constitute the latexin family of potential inhibitors. 15738388 1 43 theme protein 107:113 arg1 inhibitor 115:123 The only endogenous protein inhibitor 87:123 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) 87:165 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 1 43 theme protein 107:113 arg1 latexin 170:176 latexin 170:176 latexin 170:176 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 10 44 theme funnel-like 1297:1307 arg1 access 1309:1314 the funnel-like access 1293:1314 the funnel-like access to the protease active site 1293:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 12 45 theme potential 1575:1583 arg1 inhibitors 1585:1594 potential inhibitors 1575:1594 potential inhibitors 1575:1594 They are proposed to constitute the latexin family of potential inhibitors. 15738388 10 46 theme access 1309:1314 arg1 border 1283:1288 the border 1279:1288 the border of the funnel-like access to the protease active site 1279:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 13 47 theme inhibitors 1702:1711 arg1 counterparts 1679:1690 the counterparts 1675:1690 the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases 1675:1761 Because they are ubiquitous, latexins could represent for vertebrate A/B MCPs the counterparts of tissue inhibitors of metalloproteases for matrix metalloproteinases. 15738388 7 48 theme contact 899:905 arg1 surface 907:913 a large contact surface 891:913 a large contact surface 891:913 The complex occludes a large contact surface but makes rather few contacts, despite a nanomolar inhibition constant. 15738388 4 49 theme Human 520:524 arg1 latexin 526:532 Human latexin 520:532 Human latexin 520:532 Human latexin consists of two topologically equivalent subdomains, reminiscent of cystatins, consisting of an alpha-helix enveloped by a curved beta-sheet. 15738388 9 50 theme MCPs 1182:1185 arg1 MCPs 1208:1211 MCPs and invertebrate A/B MCPs 1182:1211 MCPs and invertebrate A/B MCPs 1182:1211 In contrast, modeling studies reveal why the N/E subfamily of MCPs and invertebrate A/B MCPs are not inhibited. 15738388 2 51 theme endogenous 240:249 arg1 inhibitor 268:276 alias endogenous carboxypeptidase inhibitor 234:276 alias endogenous carboxypeptidase inhibitor 234:276 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 2 51 theme endogenous 240:249 arg1 Latexin 225:231 Latexin 225:231 Latexin 225:231 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 0 52 with Structure 0:8 arg1 latexin 78:84 latexin 78:84 latexin 78:84 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 0 52 with Structure 0:8 arg1 inhibitor 67:75 its endogenous protein inhibitor 44:75 its endogenous protein inhibitor 44:75 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 3 53 theme subfamily 443:451 arg1 hCPA4 414:418 hCPA4 414:418 hCPA4 414:418 hCPA4 is a member of the A/B subfamily of MCPs and displays the characteristic alpha/beta-hydrolase fold. 15738388 3 53 theme subfamily 443:451 arg1 member 425:430 a member 423:430 a member of the A/B subfamily of MCPs 423:459 hCPA4 is a member of the A/B subfamily of MCPs and displays the characteristic alpha/beta-hydrolase fold. 15738388 10 54 theme active 1332:1337 arg1 site 1339:1342 the protease active site 1319:1342 the protease active site 1319:1342 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 4 55 theme cystatins 602:610 arg1 reminiscent 587:597 reminiscent 587:597 reminiscent of cystatins 587:610 Human latexin consists of two topologically equivalent subdomains, reminiscent of cystatins, consisting of an alpha-helix enveloped by a curved beta-sheet. 15738388 4 55 theme cystatins 602:610 arg1 subdomains 575:584 two topologically equivalent subdomains 546:584 two topologically equivalent subdomains 546:584 Human latexin consists of two topologically equivalent subdomains, reminiscent of cystatins, consisting of an alpha-helix enveloped by a curved beta-sheet. 15738388 8 56 theme species 1102:1108 arg1 barriers 1110:1117 species barriers 1102:1117 species barriers 1102:1117 This low specificity explains the flexibility of latexin in inhibiting all vertebrate A/B MCPs tested, even across species barriers. 15738388 2 57 theme alias 234:238 arg1 inhibitor 268:276 alias endogenous carboxypeptidase inhibitor 234:276 alias endogenous carboxypeptidase inhibitor 234:276 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 2 57 theme alias 234:238 arg1 Latexin 225:231 Latexin 225:231 Latexin 225:231 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 9 58 theme modeling 1133:1140 arg1 studies 1142:1148 modeling studies 1133:1148 modeling studies 1133:1148 In contrast, modeling studies reveal why the N/E subfamily of MCPs and invertebrate A/B MCPs are not inhibited. 15738388 1 59 theme rat 214:216 arg1 brain 218:222 the rat brain 210:222 the rat brain 210:222 The only endogenous protein inhibitor known for metallocarboxypeptidases (MCPs) is latexin, a 25-kDa protein discovered in the rat brain. 15738388 9 60 theme MCPs 1208:1211 arg1 subfamily 1169:1177 the N/E subfamily 1161:1177 the N/E subfamily of MCPs and invertebrate A/B MCPs 1161:1211 In contrast, modeling studies reveal why the N/E subfamily of MCPs and invertebrate A/B MCPs are not inhibited. 15738388 2 61 theme prostate 339:346 arg1 cells 355:359 prostate cancer cells 339:359 prostate cancer cells 339:359 Latexin, alias endogenous carboxypeptidase inhibitor, inhibits human CPA4 (hCPA4), whose expression is induced in prostate cancer cells after treatment with histone deacetylase inhibitors. 15738388 7 62 theme large 893:897 arg1 surface 907:913 a large contact surface 891:913 a large contact surface 891:913 The complex occludes a large contact surface but makes rather few contacts, despite a nanomolar inhibition constant. 15738388 10 63 theme complex 1351:1357 arg1 formation 1359:1367 complex formation 1351:1367 complex formation 1351:1367 Major differences in the loop segments shaping the border of the funnel-like access to the protease active site impede complex formation with latexin. 15738388 0 64 theme protein 59:65 arg1 latexin 78:84 latexin 78:84 latexin 78:84 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 0 64 theme protein 59:65 arg1 inhibitor 67:75 its endogenous protein inhibitor 44:75 its endogenous protein inhibitor 44:75 Structure of human carboxypeptidase A4 with its endogenous protein inhibitor, latexin. 15738388 7 65 theme nanomolar 956:964 arg1 constant 977:984 a nanomolar inhibition constant 954:984 a nanomolar inhibition constant 954:984 The complex occludes a large contact surface but makes rather few contacts, despite a nanomolar inhibition constant. 15738388 3 66 theme alpha/beta-hydrolase 493:512 arg1 fold 514:517 the characteristic alpha/beta-hydrolase fold 474:517 the characteristic alpha/beta-hydrolase fold 474:517 hCPA4 is a member of the A/B subfamily of MCPs and displays the characteristic alpha/beta-hydrolase fold. 16091843 7 0 theme PD 1080:1081 arg1 inhibition 1083:1092 PD inhibition 1080:1092 PD inhibition 1080:1092 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 2 1 theme related 382:388 arg1 zymogens 414:421 related metallocarboxypeptidase zymogens 382:421 related metallocarboxypeptidase zymogens 382:421 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 6 2 theme inhibitory 882:891 arg1 evocative 901:909 evocative 901:909 evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4 901:1001 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 6 2 theme inhibitory 882:891 arg1 mode 893:896 Such an inhibitory mode 874:896 Such an inhibitory mode 874:896 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 1 3 theme advanced 163:170 arg1 stages 172:177 advanced stages 163:177 advanced stages of prostate carcinoma 163:199 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 6 4 theme latexin 969:975 arg1 structure 936:944 the recently reported structure 914:944 the recently reported structure of the human inhibitor latexin 914:975 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 5 5 theme funnel 723:728 arg1 border 708:713 The border 704:713 The border of this funnel 704:728 The border of this funnel is shaped by loops, which are responsible for the interaction with the PD, characterised by a large interface area and relatively few contacts. 16091843 3 6 theme spherical 580:588 arg1 cone 590:593 a spherical cone 578:593 a spherical cone 578:593 The protease moiety recalls a sphere, out of which a spherical cone has been cut. 16091843 6 7 theme inhibitor 959:967 arg1 latexin 969:975 the human inhibitor latexin 949:975 the human inhibitor latexin 949:975 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 0 8 theme zymogen 95:101 arg1 state 103:107 the zymogen state 91:107 the zymogen state 91:107 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 1 9 with Treatment 150:158 arg1 inhibitors 226:235 histone-deacetylase inhibitors 206:235 histone-deacetylase inhibitors 206:235 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 5 10 with interaction 780:790 arg1 PD 801:802 the PD 797:802 the PD 797:802 The border of this funnel is shaped by loops, which are responsible for the interaction with the PD, characterised by a large interface area and relatively few contacts. 16091843 5 11 theme interface 830:838 arg1 area 840:843 a large interface area 822:843 a large interface area 822:843 The border of this funnel is shaped by loops, which are responsible for the interaction with the PD, characterised by a large interface area and relatively few contacts. 16091843 0 12 from mode 53:56 arg1 state 103:107 the zymogen state 91:107 the zymogen state 91:107 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 8 13 theme sheet 1194:1198 arg1 part 1179:1182 the central part 1167:1182 the central part of a beta sheet 1167:1198 In both cases, active-site blocking relies mainly on a loop provided by the central part of a beta sheet. 16091843 2 14 theme three-dimensional 302:318 arg1 structure 320:328 The three-dimensional structure 298:328 The three-dimensional structure of hPCPA4 298:338 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 8 15 theme beta 1189:1192 arg1 sheet 1194:1198 a beta sheet 1187:1198 a beta sheet 1187:1198 In both cases, active-site blocking relies mainly on a loop provided by the central part of a beta sheet. 16091843 1 16 theme stages 172:177 arg1 Treatment 150:158 Treatment 150:158 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors 150:235 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 6 17 theme human 953:957 arg1 latexin 969:975 the human inhibitor latexin 949:975 the human inhibitor latexin 949:975 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 0 18 theme molecular 9:17 arg1 comparison 19:28 Detailed molecular comparison 0:28 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 0:148 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 7 19 theme contacting 1013:1022 arg1 similar 1048:1054 similar 1048:1054 similar 1048:1054 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 7 19 theme contacting 1013:1022 arg1 structure 1024:1032 The main contacting structure 1004:1032 The main contacting structure of latexin 1004:1043 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 1 20 theme prostate 182:189 arg1 carcinoma 191:199 prostate carcinoma 182:199 prostate carcinoma 182:199 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 0 21 theme Detailed 0:7 arg1 comparison 19:28 Detailed molecular comparison 0:28 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 0:148 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 7 22 theme latexin 1037:1043 arg1 similar 1048:1054 similar 1048:1054 similar 1048:1054 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 7 22 theme latexin 1037:1043 arg1 structure 1024:1032 The main contacting structure 1004:1032 The main contacting structure of latexin 1004:1043 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 1 23 theme carcinoma 191:199 arg1 stages 172:177 advanced stages 163:177 advanced stages of prostate carcinoma 163:199 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 0 24 theme endogenous 120:129 arg1 latexin 141:147 the endogenous inhibitor latexin 116:147 the endogenous inhibitor latexin 116:147 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 6 25 theme structure 936:944 arg1 evocative 901:909 evocative 901:909 evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4 901:1001 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 6 25 theme structure 936:944 arg1 mode 893:896 Such an inhibitory mode 874:896 Such an inhibitory mode 874:896 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 2 26 theme hPCPA4 333:338 arg1 structure 320:328 The three-dimensional structure 298:328 The three-dimensional structure of hPCPA4 298:338 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 1 27 theme histone-deacetylase 206:224 arg1 inhibitors 226:235 histone-deacetylase inhibitors 206:235 histone-deacetylase inhibitors 206:235 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 0 28 theme inhibition 42:51 arg1 mode 53:56 the inhibition mode 38:56 the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin 38:147 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 6 29 theme reported 927:934 arg1 structure 936:944 the recently reported structure 914:944 the recently reported structure of the human inhibitor latexin 914:975 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 6 30 with complex 984:990 arg1 hCPA4 997:1001 hCPA4 997:1001 hCPA4 997:1001 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 2 31 theme active-enzyme 463:475 arg1 hCPA4 485:489 hCPA4 485:489 hCPA4 485:489 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 31 theme active-enzyme 463:475 arg1 moiety 477:482 a preformed alpha/beta/-hydrolase active-enzyme moiety 429:482 a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) 429:490 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 32 theme preformed 431:439 arg1 hCPA4 485:489 hCPA4 485:489 hCPA4 485:489 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 32 theme preformed 431:439 arg1 moiety 477:482 a preformed alpha/beta/-hydrolase active-enzyme moiety 429:482 a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) 429:490 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 0 33 theme inhibitor 131:139 arg1 latexin 141:147 the endogenous inhibitor latexin 116:147 the endogenous inhibitor latexin 116:147 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 4 34 theme funnel-like 627:637 arg1 structure 639:647 a funnel-like structure 625:647 a funnel-like structure 625:647 This results in a funnel-like structure, at the bottom of which the active-site cleft resides. 16091843 2 35 theme alpha/beta/-hydrolase 441:461 arg1 hCPA4 485:489 hCPA4 485:489 hCPA4 485:489 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 35 theme alpha/beta/-hydrolase 441:461 arg1 moiety 477:482 a preformed alpha/beta/-hydrolase active-enzyme moiety 429:482 a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) 429:490 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 8 36 theme central 1171:1177 arg1 part 1179:1182 the central part 1167:1182 the central part of a beta sheet 1167:1198 In both cases, active-site blocking relies mainly on a loop provided by the central part of a beta sheet. 16091843 5 37 theme few 860:862 arg1 contacts 864:871 relatively few contacts 849:871 relatively few contacts 849:871 The border of this funnel is shaped by loops, which are responsible for the interaction with the PD, characterised by a large interface area and relatively few contacts. 16091843 7 38 theme main 1008:1011 arg1 similar 1048:1054 similar 1048:1054 similar 1048:1054 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 7 38 theme main 1008:1011 arg1 structure 1024:1032 The main contacting structure 1004:1032 The main contacting structure of latexin 1004:1043 The main contacting structure of latexin is similar to the one employed for PD inhibition. 16091843 2 39 theme inhibiting 499:508 arg1 pro-domain 510:519 an inhibiting pro-domain 496:519 an inhibiting pro-domain (PD) 496:524 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 39 theme inhibiting 499:508 arg1 PD 522:523 PD 522:523 PD 522:523 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 6 40 from evocative 901:909 arg1 complex 984:990 its complex 980:990 its complex with hCPA4 980:1001 Such an inhibitory mode is evocative of the recently reported structure of the human inhibitor latexin in its complex with hCPA4. 16091843 3 41 theme protease 531:538 arg1 moiety 540:545 The protease moiety 527:545 The protease moiety 527:545 The protease moiety recalls a sphere, out of which a spherical cone has been cut. 16091843 8 42 theme active-site 1110:1120 arg1 blocking 1122:1129 active-site blocking 1110:1129 active-site blocking 1110:1129 In both cases, active-site blocking relies mainly on a loop provided by the central part of a beta sheet. 16091843 2 43 with zymogens 414:421 arg1 pro-domain 510:519 an inhibiting pro-domain 496:519 an inhibiting pro-domain (PD) 496:524 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 43 with zymogens 414:421 arg1 hCPA4 485:489 hCPA4 485:489 hCPA4 485:489 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 43 with zymogens 414:421 arg1 moiety 477:482 a preformed alpha/beta/-hydrolase active-enzyme moiety 429:482 a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) 429:490 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 2 43 with zymogens 414:421 arg1 PD 522:523 PD 522:523 PD 522:523 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 4 44 theme active-site 677:687 arg1 cleft 689:693 the active-site cleft 673:693 the active-site cleft 673:693 This results in a funnel-like structure, at the bottom of which the active-site cleft resides. 16091843 0 45 theme carboxypeptidases 70:86 arg1 mode 53:56 the inhibition mode 38:56 the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin 38:147 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 5 46 theme large 824:828 arg1 area 840:843 a large interface area 822:843 a large interface area 822:843 The border of this funnel is shaped by loops, which are responsible for the interaction with the PD, characterised by a large interface area and relatively few contacts. 16091843 2 47 theme zymogens 414:421 arg1 features 370:377 the features 366:377 the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD) 366:524 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 1 48 theme human 259:263 arg1 hPCPA4 289:294 hPCPA4 289:294 hPCPA4 289:294 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 1 48 theme human 259:263 arg1 procarboxypeptidase-A4 265:286 human procarboxypeptidase-A4 259:286 human procarboxypeptidase-A4 (hPCPA4) 259:295 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 16091843 0 49 theme A/B-type 61:68 arg1 carboxypeptidases 70:86 A/B-type carboxypeptidases 61:86 A/B-type carboxypeptidases 61:86 Detailed molecular comparison between the inhibition mode of A/B-type carboxypeptidases in the zymogen state and by the endogenous inhibitor latexin. 16091843 2 50 theme metallocarboxypeptidase 390:412 arg1 zymogens 414:421 related metallocarboxypeptidase zymogens 382:421 related metallocarboxypeptidase zymogens 382:421 The three-dimensional structure of hPCPA4 has been solved and shows the features of related metallocarboxypeptidase zymogens, with a preformed alpha/beta/-hydrolase active-enzyme moiety (hCPA4) and an inhibiting pro-domain (PD). 16091843 1 51 theme procarboxypeptidase-A4 265:286 arg1 expression 245:254 expression 245:254 expression of human procarboxypeptidase-A4 (hPCPA4) 245:295 Treatment of advanced stages of prostate carcinoma with histone-deacetylase inhibitors entails expression of human procarboxypeptidase-A4 (hPCPA4). 32079768 3 0 from network 630:636 arg1 core 654:657 the receptor core 641:657 the receptor core 641:657 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 4 1 theme signaling 796:804 arg1 Divergences 709:719 Divergences 709:719 Divergences 709:719 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 1 theme signaling 796:804 arg1 switch 806:811 a biased signaling switch 787:811 a biased signaling switch 787:811 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 2 theme biased 789:794 arg1 Divergences 709:719 Divergences 709:719 Divergences 709:719 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 2 theme biased 789:794 arg1 switch 806:811 a biased signaling switch 787:811 a biased signaling switch 787:811 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 3 theme family 730:735 arg1 consensus 737:745 the family consensus 726:745 the family consensus 726:745 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 4 theme protein 983:989 arg1 signaling 991:999 heterotrimeric Gq protein signaling 965:999 β-arrestin but not heterotrimeric Gq protein signaling 946:999 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 5 dep GPCRs 856:860 arg1 such 863:866 such 863:866 such 863:866 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 3 6 theme receptor 645:652 arg1 core 654:657 the receptor core 641:657 the receptor core 641:657 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 3 7 theme more-substantial 522:537 arg1 rearrangements 539:552 more-substantial rearrangements 522:552 more-substantial rearrangements 522:552 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 4 8 from Divergences 709:719 arg1 region 755:760 this region 750:760 this region 750:760 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 2 9 theme type 280:283 arg1 AT1R 297:300 AT1R 297:300 AT1R 297:300 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 9 theme type 280:283 arg1 angstroms 315:323 2.7 to 2.9 angstroms 304:323 2.7 to 2.9 angstroms 304:323 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 9 theme type 280:283 arg1 receptor 287:294 angiotensin II type 1 receptor 265:294 angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs 265:475 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 4 10 theme chemokine 871:879 arg1 receptors 881:889 chemokine receptors 871:889 chemokine receptors 871:889 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 3 11 theme sodium-binding 674:687 arg1 site 689:692 a sodium-binding site 672:692 a sodium-binding site in most GPCRs 672:706 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 3 12 from site 689:692 arg1 GPCRs 702:706 most GPCRs 697:706 most GPCRs 697:706 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 3 13 theme other 492:496 arg1 ligands 498:504 other ligands 492:504 other ligands 492:504 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 2 14 theme divergent 354:362 arg1 profiles 369:376 divergent bias profiles 354:376 divergent bias profiles 354:376 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 15 theme angiotensin 265:275 arg1 AT1R 297:300 AT1R 297:300 AT1R 297:300 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 15 theme angiotensin 265:275 arg1 angstroms 315:323 2.7 to 2.9 angstroms 304:323 2.7 to 2.9 angstroms 304:323 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 15 theme angiotensin 265:275 arg1 receptor 287:294 angiotensin II type 1 receptor 265:294 angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs 265:475 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 4 16 theme Gq 980:981 arg1 signaling 991:999 heterotrimeric Gq protein signaling 965:999 β-arrestin but not heterotrimeric Gq protein signaling 946:999 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 2 17 theme agonist 403:409 arg1 angiotensin 411:421 the balanced endogenous agonist angiotensin II 379:424 the balanced endogenous agonist angiotensin II (AngII) 379:432 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 17 theme agonist 403:409 arg1 AngII 427:431 AngII 427:431 AngII 427:431 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 3 18 theme most 697:700 arg1 GPCRs 702:706 most GPCRs 697:706 most GPCRs 697:706 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 1 19 theme Biased 96:101 arg1 agonists 103:110 Biased agonists 96:110 Biased agonists of G protein-coupled receptors (GPCRs) 96:149 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 1 20 theme downstream 187:196 arg1 pathways 208:215 downstream signaling pathways 187:215 downstream signaling pathways 187:215 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 2 21 theme endogenous 392:401 arg1 angiotensin 411:421 the balanced endogenous agonist angiotensin II 379:424 the balanced endogenous agonist angiotensin II (AngII) 379:432 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 21 theme endogenous 392:401 arg1 AngII 427:431 AngII 427:431 AngII 427:431 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 4 22 theme certain 842:848 arg1 GPCRs 856:860 certain other GPCRs 842:860 certain other GPCRs (such as chemokine receptors) 842:890 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 2 23 dep 2.9 311:313 arg1 to 308:309 to 308:309 to 308:309 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 1 24 theme signaling 198:206 arg1 pathways 208:215 downstream signaling pathways 187:215 downstream signaling pathways 187:215 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 2 25 theme balanced 383:390 arg1 angiotensin 411:421 the balanced endogenous agonist angiotensin II 379:424 the balanced endogenous agonist angiotensin II (AngII) 379:432 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 25 theme balanced 383:390 arg1 AngII 427:431 AngII 427:431 AngII 427:431 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 26 theme receptor 287:294 arg1 structures 251:260 crystal structures 243:260 crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs 243:475 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 1 27 theme pathways 208:215 arg1 subset 177:182 a subset 175:182 a subset of downstream signaling pathways 175:215 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 4 28 theme other 850:854 arg1 GPCRs 856:860 certain other GPCRs 842:860 certain other GPCRs (such as chemokine receptors) 842:890 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 2 29 theme bias 364:367 arg1 profiles 369:376 divergent bias profiles 354:376 divergent bias profiles 354:376 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 0 30 theme active 60:65 arg1 conformations 67:79 structurally distinct active conformations 38:79 structurally distinct active conformations 38:79 Angiotensin and biased analogs induce structurally distinct active conformations within a GPCR. 32079768 0 31 theme distinct 51:58 arg1 conformations 67:79 structurally distinct active conformations 38:79 structurally distinct active conformations 38:79 Angiotensin and biased analogs induce structurally distinct active conformations within a GPCR. 32079768 1 32 theme protein-coupled 117:131 arg1 receptors 133:141 G protein-coupled receptors 115:141 G protein-coupled receptors (GPCRs) 115:149 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 1 32 theme protein-coupled 117:131 arg1 GPCRs 144:148 GPCRs 144:148 GPCRs 144:148 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 3 33 theme ligand-binding 584:597 arg1 pocket 599:604 the ligand-binding pocket 580:604 the ligand-binding pocket 580:604 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 2 34 dep ligands 341:347 arg1 angiotensin 411:421 the balanced endogenous agonist angiotensin II 379:424 the balanced endogenous agonist angiotensin II (AngII) 379:432 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 34 dep ligands 341:347 arg1 AngII 427:431 AngII 427:431 AngII 427:431 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 34 dep ligands 341:347 arg1 analogs 469:475 two strongly β-arrestin-biased analogs 438:475 two strongly β-arrestin-biased analogs 438:475 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 1 35 theme receptors 133:141 arg1 agonists 103:110 Biased agonists 96:110 Biased agonists of G protein-coupled receptors (GPCRs) 96:149 Biased agonists of G protein-coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. 32079768 3 36 theme key 620:622 arg1 network 630:636 a key polar network 618:636 a key polar network 618:636 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 3 37 theme pocket 599:604 arg1 bottom 570:575 the bottom 566:575 the bottom of the ligand-binding pocket 566:604 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 2 38 theme β-arrestin-biased 451:467 arg1 analogs 469:475 two strongly β-arrestin-biased analogs 438:475 two strongly β-arrestin-biased analogs 438:475 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 3 39 theme polar 624:628 arg1 network 630:636 a key polar network 618:636 a key polar network 618:636 Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. 32079768 2 40 theme crystal 243:249 arg1 structures 251:260 crystal structures 243:260 crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs 243:475 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 41 attach present 235:241 arg2 we 232:233 we 232:233 we 232:233 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 41 attach present 235:241 arg1 work 226:229 this work 221:229 this work 221:229 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 2 42 with ligands 341:347 arg1 profiles 369:376 divergent bias profiles 354:376 divergent bias profiles 354:376 In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin-biased analogs. 32079768 0 43 theme biased 16:21 arg1 analogs 23:29 biased analogs 16:29 biased analogs 16:29 Angiotensin and biased analogs induce structurally distinct active conformations within a GPCR. 32079768 4 44 from consensus 737:745 arg1 Divergences 709:719 Divergences 709:719 Divergences 709:719 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 44 from consensus 737:745 arg1 switch 806:811 a biased signaling switch 787:811 a biased signaling switch 787:811 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 32079768 4 45 theme heterotrimeric 965:978 arg1 signaling 991:999 heterotrimeric Gq protein signaling 965:999 β-arrestin but not heterotrimeric Gq protein signaling 946:999 Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling. 29748286 0 0 theme B. 85:86 arg1 infection 110:118 B. Clostridium difficile infection 85:118 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 1 1 theme FZD 314:316 arg1 family 319:324 the frizzled (FZD) family 300:324 the frizzled (FZD) family of Wnt receptors 300:341 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 2 2 theme endogenous 543:552 arg1 acid 570:573 an endogenous FZD-bound fatty acid 540:573 an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 540:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 4 3 from lipid 720:724 arg1 place 729:733 place 729:733 place 729:733 TcdB binding locks the lipid in place, preventing Wnt from engaging FZDs and signaling. 29748286 3 4 theme binding 641:647 arg1 site 649:652 the binding site 637:652 the binding site for Wnt-adducted palmitoleic acid in FZDs 637:694 This lipid occupies the binding site for Wnt-adducted palmitoleic acid in FZDs. 29748286 0 5 theme developed 182:190 arg1 countries 192:200 developed countries 182:200 developed countries 182:200 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 2 6 theme TcdB 429:432 arg1 fragment 434:441 a TcdB fragment 427:441 a TcdB fragment 427:441 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 5 7 theme fatty 826:830 arg1 acids 832:836 fatty acids 826:836 fatty acids 826:836 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 0 8 theme difficile 100:108 arg1 infection 110:118 B. Clostridium difficile infection 85:118 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 0 9 theme Clostridium 88:98 arg1 infection 110:118 B. Clostridium difficile infection 85:118 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 3 10 theme Wnt-adducted 658:669 arg1 acid 683:686 Wnt-adducted palmitoleic acid 658:686 Wnt-adducted palmitoleic acid in FZDs 658:694 This lipid occupies the binding site for Wnt-adducted palmitoleic acid in FZDs. 29748286 4 11 theme TcdB 697:700 arg1 binding 702:708 TcdB binding 697:708 TcdB binding 697:708 TcdB binding locks the lipid in place, preventing Wnt from engaging FZDs and signaling. 29748286 2 12 from resolution 514:523 arg1 domain 477:482 the cysteine-rich domain 459:482 the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 459:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 12 from resolution 514:523 arg1 FZD2 493:496 human FZD2 487:496 human FZD2 487:496 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 1 13 theme Wnt 329:331 arg1 receptors 333:341 Wnt receptors 329:341 Wnt receptors 329:341 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 3 14 from acid 683:686 arg1 FZDs 691:694 FZDs 691:694 FZDs 691:694 This lipid occupies the binding site for Wnt-adducted palmitoleic acid in FZDs. 29748286 5 15 from role 818:821 arg1 pathogenesis 859:870 FZD-mediated TcdB pathogenesis 841:870 FZD-mediated TcdB pathogenesis 841:870 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 5 16 theme Wnt 907:909 arg1 signaling 911:919 Wnt signaling 907:919 Wnt signaling 907:919 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 1 17 theme receptors 333:341 arg1 family 319:324 the frizzled (FZD) family 300:324 the frizzled (FZD) family of Wnt receptors 300:341 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 2 18 from structure 414:422 arg1 complex 446:452 complex 446:452 complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 446:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 19 theme fatty 564:568 arg1 acid 570:573 an endogenous FZD-bound fatty acid 540:573 an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 540:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 20 with complex 446:452 arg1 domain 477:482 the cysteine-rich domain 459:482 the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 459:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 20 with complex 446:452 arg1 FZD2 493:496 human FZD2 487:496 human FZD2 487:496 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 3 21 theme palmitoleic 671:681 arg1 acid 683:686 Wnt-adducted palmitoleic acid 658:686 Wnt-adducted palmitoleic acid in FZDs 658:694 This lipid occupies the binding site for Wnt-adducted palmitoleic acid in FZDs. 29748286 0 22 theme Structural 0:9 arg1 cause 139:143 the most common cause 123:143 the most common cause of antibiotic-associated diarrhea in developed countries 123:200 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 0 22 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection 0:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 2 23 theme FZD-bound 554:562 arg1 acid 570:573 an endogenous FZD-bound fatty acid 540:573 an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 540:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 0 24 dep toxin 79:83 arg1 infection 110:118 B. Clostridium difficile infection 85:118 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 1 25 theme colonic 268:274 arg1 epithelia 276:284 colonic epithelia 268:284 colonic epithelia 268:284 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 2 26 theme FZD2 493:496 arg1 domain 477:482 the cysteine-rich domain 459:482 the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 459:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 26 theme FZD2 493:496 arg1 FZD2 493:496 human FZD2 487:496 human FZD2 487:496 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 27 theme TcdB 603:606 arg1 binding 608:614 TcdB binding 603:614 TcdB binding 603:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 1 28 theme major 207:211 arg1 B 250:250 C. difficile toxin B 231:250 C. difficile toxin B (TcdB) 231:257 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 28 theme major 207:211 arg1 factor 223:228 The major virulence factor 203:228 The major virulence factor 203:228 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 2 29 theme human 487:491 arg1 FZD2 493:496 human FZD2 487:496 human FZD2 487:496 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 1 30 theme virulence 213:221 arg1 B 250:250 C. difficile toxin B 231:250 C. difficile toxin B (TcdB) 231:257 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 30 theme virulence 213:221 arg1 factor 223:228 The major virulence factor 203:228 The major virulence factor 203:228 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 0 31 theme frizzled 36:43 arg1 proteins 45:52 frizzled proteins 36:52 frizzled proteins 36:52 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 0 32 theme common 132:137 arg1 cause 139:143 the most common cause 123:143 the most common cause of antibiotic-associated diarrhea in developed countries 123:200 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 0 32 theme common 132:137 arg1 basis 11:15 Structural basis 0:15 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection 0:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 2 33 theme crystal 406:412 arg1 structure 414:422 the crystal structure 402:422 the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 402:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 34 theme cysteine-rich 463:475 arg1 domain 477:482 the cysteine-rich domain 459:482 the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 459:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 2 34 theme cysteine-rich 463:475 arg1 FZD2 493:496 human FZD2 487:496 human FZD2 487:496 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 0 35 theme antibiotic-associated 148:168 arg1 diarrhea 170:177 antibiotic-associated diarrhea 148:177 antibiotic-associated diarrhea 148:177 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 2 36 theme fragment 434:441 arg1 structure 414:422 the crystal structure 402:422 the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding 402:614 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 5 37 theme acids 832:836 arg1 role 818:821 a central role 808:821 a central role of fatty acids in FZD-mediated TcdB pathogenesis 808:870 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 0 38 theme proteins 45:52 arg1 recognition 21:31 recognition 21:31 recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection 21:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 1 39 theme frizzled 304:311 arg1 family 319:324 the frizzled (FZD) family 300:324 the frizzled (FZD) family of Wnt receptors 300:341 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 2 40 theme 2.5-angstrom 501:512 arg1 resolution 514:523 2.5-angstrom resolution 501:523 2.5-angstrom resolution 501:523 Here, we present the crystal structure of a TcdB fragment in complex with the cysteine-rich domain of human FZD2 at 2.5-angstrom resolution, which reveals an endogenous FZD-bound fatty acid acting as a co-receptor for TcdB binding. 29748286 5 41 theme FZD-mediated 841:852 arg1 pathogenesis 859:870 FZD-mediated TcdB pathogenesis 841:870 FZD-mediated TcdB pathogenesis 841:870 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 0 42 from cause 139:143 arg1 countries 192:200 developed countries 182:200 developed countries 182:200 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 0 43 theme diarrhea 170:177 arg1 cause 139:143 the most common cause 123:143 the most common cause of antibiotic-associated diarrhea in developed countries 123:200 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 0 43 theme diarrhea 170:177 arg1 basis 11:15 Structural basis 0:15 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection 0:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 5 44 theme TcdB 854:857 arg1 pathogenesis 859:870 FZD-mediated TcdB pathogenesis 841:870 FZD-mediated TcdB pathogenesis 841:870 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 1 45 theme difficile 234:242 arg1 TcdB 253:256 TcdB 253:256 TcdB 253:256 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 45 theme difficile 234:242 arg1 B 250:250 C. difficile toxin B 231:250 C. difficile toxin B (TcdB) 231:257 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 45 theme difficile 234:242 arg1 factor 223:228 The major virulence factor 203:228 The major virulence factor 203:228 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 0 46 theme difficile 69:77 arg1 toxin 79:83 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 5 47 theme central 810:816 arg1 role 818:821 a central role 808:821 a central role of fatty acids in FZD-mediated TcdB pathogenesis 808:870 Our findings establish a central role of fatty acids in FZD-mediated TcdB pathogenesis and suggest strategies to modulate Wnt signaling. 29748286 1 48 theme toxin 244:248 arg1 TcdB 253:256 TcdB 253:256 TcdB 253:256 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 48 theme toxin 244:248 arg1 B 250:250 C. difficile toxin B 231:250 C. difficile toxin B (TcdB) 231:257 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 48 theme toxin 244:248 arg1 factor 223:228 The major virulence factor 203:228 The major virulence factor 203:228 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 0 49 theme Clostridium 57:67 arg1 toxin 79:83 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Clostridium difficile toxin B. Clostridium difficile infection 57:118 Structural basis for recognition of frizzled proteins by Clostridium difficile toxin B. Clostridium difficile infection is the most common cause of antibiotic-associated diarrhea in developed countries. 29748286 1 50 theme C. 231:232 arg1 TcdB 253:256 TcdB 253:256 TcdB 253:256 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 50 theme C. 231:232 arg1 B 250:250 C. difficile toxin B 231:250 C. difficile toxin B (TcdB) 231:257 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 29748286 1 50 theme C. 231:232 arg1 factor 223:228 The major virulence factor 203:228 The major virulence factor 203:228 The major virulence factor, C. difficile toxin B (TcdB), targets colonic epithelia by binding to the frizzled (FZD) family of Wnt receptors, but how TcdB recognizes FZDs is unclear. 12169660 0 0 theme multistep 81:89 arg1 mechanism 102:110 a multistep allosteric mechanism 79:110 a multistep allosteric mechanism 79:110 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 6 1 theme HCII 1026:1029 arg1 structure 1013:1021 The native structure 1002:1021 The native structure of HCII 1002:1029 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 6 2 dep resembles 1031:1039 arg1 whereas 1133:1139 whereas 1133:1139 whereas 1133:1139 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 6 3 theme allosteric 1110:1119 arg1 activation 1121:1130 allosteric activation 1110:1130 allosteric activation 1110:1130 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 1 4 theme fibrin 167:172 arg1 clot 174:177 a fibrin clot 165:177 a fibrin clot 165:177 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 5 5 theme sequestered 930:940 arg1 tail 966:969 a sequestered hirudin-like N-terminal tail 928:969 a sequestered hirudin-like N-terminal tail for interaction with thrombin 928:999 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 7 6 theme beta-sheet 1366:1375 arg1 expansion 1341:1349 expansion 1341:1349 expansion 1341:1349 Together, these structures reveal a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII. 12169660 7 6 theme beta-sheet 1366:1375 arg1 contraction 1325:1335 sequential contraction 1314:1335 sequential contraction 1314:1335 Together, these structures reveal a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII. 12169660 5 7 theme tail 966:969 arg1 release 917:923 the release 913:923 the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin 913:999 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 1 8 theme unique 248:253 arg1 mechanism 276:284 a unique beta-sheet expansion mechanism 246:284 a unique beta-sheet expansion mechanism to trap 246:292 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 0 9 theme allosteric 91:100 arg1 mechanism 102:110 a multistep allosteric mechanism 79:110 a multistep allosteric mechanism 79:110 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 6 10 theme molecular 1202:1210 arg1 basis 1212:1216 the molecular basis 1198:1216 the molecular basis of allostery 1198:1229 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 7 11 theme multistep 1268:1276 arg1 mechanism 1289:1297 a multistep allosteric mechanism 1266:1297 a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII 1266:1383 Together, these structures reveal a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII. 12169660 4 12 theme cofactor 768:775 arg1 structures 710:719 the structures 706:719 the structures of native and S195A thrombin-complexed heparin cofactor II (HCII) 706:785 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 1 13 theme beta-sheet 255:264 arg1 mechanism 276:284 a unique beta-sheet expansion mechanism 246:284 a unique beta-sheet expansion mechanism to trap 246:292 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 2 14 from role 435:438 arg1 mechanism 474:482 their mechanism 468:482 their mechanism 468:482 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 7 15 theme HCII 1380:1383 arg1 beta-sheet 1366:1375 beta-sheet 1366:1375 beta-sheet 1366:1375 Together, these structures reveal a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII. 12169660 2 16 theme protease 378:385 arg1 inhibitors 387:396 serine protease inhibitors 371:396 serine protease inhibitors 371:396 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 5 17 with interaction 975:985 arg1 thrombin 992:999 thrombin 992:999 thrombin 992:999 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 0 18 theme thrombin-complexed 33:50 arg1 cofactor 60:67 native and thrombin-complexed heparin cofactor II 22:70 native and thrombin-complexed heparin cofactor II 22:70 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 3 19 theme protease 506:513 arg1 inhibition 515:524 protease inhibition 506:524 protease inhibition 506:524 It now is clear that protease inhibition depends entirely on rapid serpin beta-sheet expansion after proteolytic attack. 12169660 2 20 theme serine 371:376 arg1 inhibitors 387:396 serine protease inhibitors 371:396 serine protease inhibitors 371:396 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 6 21 theme complex 1182:1188 arg1 structure 1145:1153 the structure 1141:1153 the structure of the S195A thrombin-HCII complex 1141:1188 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 3 22 theme rapid 546:550 arg1 expansion 570:578 rapid serpin beta-sheet expansion 546:578 rapid serpin beta-sheet expansion after proteolytic attack 546:603 It now is clear that protease inhibition depends entirely on rapid serpin beta-sheet expansion after proteolytic attack. 12169660 4 23 theme serpins 674:680 arg1 mobility 662:669 the conformational mobility 643:669 the conformational mobility of serpins 643:680 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 0 24 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of native and thrombin-complexed heparin cofactor II 0:70 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 1 25 theme expansion 266:274 arg1 mechanism 276:284 a unique beta-sheet expansion mechanism 246:284 a unique beta-sheet expansion mechanism to trap 246:292 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 5 26 theme cascade 850:856 arg1 protease 822:829 the final protease 812:829 the final protease of the coagulation cascade 812:856 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 5 26 theme cascade 850:856 arg1 thrombin 802:809 thrombin 802:809 thrombin 802:809 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 3 27 theme serpin 552:557 arg1 expansion 570:578 rapid serpin beta-sheet expansion 546:578 rapid serpin beta-sheet expansion after proteolytic attack 546:603 It now is clear that protease inhibition depends entirely on rapid serpin beta-sheet expansion after proteolytic attack. 12169660 0 28 theme native 22:27 arg1 cofactor 60:67 native and thrombin-complexed heparin cofactor II 22:70 native and thrombin-complexed heparin cofactor II 22:70 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 6 29 theme alternative 1085:1095 arg1 mechanism 1097:1105 an alternative mechanism 1082:1105 an alternative mechanism of allosteric activation 1082:1130 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 4 30 theme heparin 760:766 arg1 cofactor 768:775 heparin cofactor II 760:778 native and S195A thrombin-complexed heparin cofactor II (HCII) 724:785 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 4 30 theme heparin 760:766 arg1 HCII 781:784 HCII 781:784 HCII 781:784 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 5 31 theme final 816:820 arg1 protease 822:829 the final protease 812:829 the final protease of the coagulation cascade 812:856 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 5 31 theme final 816:820 arg1 thrombin 802:809 thrombin 802:809 thrombin 802:809 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 5 32 theme hirudin-like 942:953 arg1 tail 966:969 a sequestered hirudin-like N-terminal tail 928:969 a sequestered hirudin-like N-terminal tail for interaction with thrombin 928:999 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 4 33 theme thrombin-complexed 741:758 arg1 cofactor 768:775 heparin cofactor II 760:778 native and S195A thrombin-complexed heparin cofactor II (HCII) 724:785 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 4 33 theme thrombin-complexed 741:758 arg1 HCII 781:784 HCII 781:784 HCII 781:784 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 6 34 theme thrombin-HCII 1168:1180 arg1 complex 1182:1188 the S195A thrombin-HCII complex 1158:1188 the S195A thrombin-HCII complex 1158:1188 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 2 35 theme change 458:463 arg1 role 435:438 the role 431:438 the role of conformational change in their mechanism 431:482 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 7 36 theme allosteric 1278:1287 arg1 mechanism 1289:1297 a multistep allosteric mechanism 1266:1297 a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII 1266:1383 Together, these structures reveal a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII. 12169660 6 37 theme native 1049:1054 arg1 antithrombin 1056:1067 native antithrombin 1049:1067 native antithrombin 1049:1067 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 2 38 theme conformational 443:456 arg1 change 458:463 conformational change 443:463 conformational change 443:463 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 6 39 theme native 1006:1011 arg1 structure 1013:1021 The native structure 1002:1021 The native structure of HCII 1002:1029 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 3 40 theme proteolytic 586:596 arg1 attack 598:603 proteolytic attack 586:603 proteolytic attack 586:603 It now is clear that protease inhibition depends entirely on rapid serpin beta-sheet expansion after proteolytic attack. 12169660 2 41 theme inhibitors 387:396 arg1 serpins 346:352 serpins 346:352 serpins 346:352 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 2 41 theme inhibitors 387:396 arg1 family 361:366 a family 359:366 a family of serine protease inhibitors 359:396 Since the discovery that serpins were a family of serine protease inhibitors there has been controversy as to the role of conformational change in their mechanism. 12169660 0 42 theme cofactor 60:67 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of native and thrombin-complexed heparin cofactor II 0:70 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 3 43 theme beta-sheet 559:568 arg1 expansion 570:578 rapid serpin beta-sheet expansion 546:578 rapid serpin beta-sheet expansion after proteolytic attack 546:603 It now is clear that protease inhibition depends entirely on rapid serpin beta-sheet expansion after proteolytic attack. 12169660 1 44 theme serine 117:122 arg1 proteases 124:132 The serine proteases 113:132 The serine proteases sequentially activated to form a fibrin clot 113:177 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 0 45 theme heparin 52:58 arg1 cofactor 60:67 native and thrombin-complexed heparin cofactor II 22:70 native and thrombin-complexed heparin cofactor II 22:70 Crystal structures of native and thrombin-complexed heparin cofactor II reveal a multistep allosteric mechanism. 12169660 6 46 theme S195A 1162:1166 arg1 complex 1182:1188 the S195A thrombin-HCII complex 1158:1188 the S195A thrombin-HCII complex 1158:1188 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 7 47 theme sequential 1314:1323 arg1 contraction 1325:1335 sequential contraction 1314:1335 sequential contraction 1314:1335 Together, these structures reveal a multistep allosteric mechanism that relies on sequential contraction and expansion of the central beta-sheet of HCII. 12169660 4 48 theme native 724:729 arg1 cofactor 768:775 heparin cofactor II 760:778 native and S195A thrombin-complexed heparin cofactor II (HCII) 724:785 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 4 48 theme native 724:729 arg1 HCII 781:784 HCII 781:784 HCII 781:784 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 5 49 theme N-terminal 955:964 arg1 tail 966:969 a sequestered hirudin-like N-terminal tail 928:969 a sequestered hirudin-like N-terminal tail for interaction with thrombin 928:999 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 1 50 theme serpin 221:226 arg1 family 228:233 the serpin family 217:233 the serpin family 217:233 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 1 51 theme family 228:233 arg1 members 206:212 members 206:212 members 206:212 The serine proteases sequentially activated to form a fibrin clot are inhibited primarily by members of the serpin family, which use a unique beta-sheet expansion mechanism to trap and destroy their targets. 12169660 5 52 theme coagulation 838:848 arg1 cascade 850:856 the coagulation cascade 834:856 the coagulation cascade 834:856 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 6 53 theme allostery 1221:1229 arg1 basis 1212:1216 the molecular basis 1198:1216 the molecular basis of allostery 1198:1229 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 12169660 5 54 theme glycosaminoglycan-dependent 864:890 arg1 manner 892:897 a glycosaminoglycan-dependent manner 862:897 a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin 862:999 HCII inhibits thrombin, the final protease of the coagulation cascade, in a glycosaminoglycan-dependent manner that involves the release of a sequestered hirudin-like N-terminal tail for interaction with thrombin. 12169660 4 55 theme conformational 647:660 arg1 mobility 662:669 the conformational mobility 643:669 the conformational mobility of serpins 643:680 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 4 56 theme regulatory 610:619 arg1 advantage 621:629 The regulatory advantage 606:629 The regulatory advantage afforded by the conformational mobility of serpins 606:680 The regulatory advantage afforded by the conformational mobility of serpins is demonstrated here by the structures of native and S195A thrombin-complexed heparin cofactor II (HCII). 12169660 6 57 theme activation 1121:1130 arg1 mechanism 1097:1105 an alternative mechanism 1082:1105 an alternative mechanism of allosteric activation 1082:1130 The native structure of HCII resembles that of native antithrombin and suggests an alternative mechanism of allosteric activation, whereas the structure of the S195A thrombin-HCII complex defines the molecular basis of allostery. 27349982 0 0 theme Domain 73:78 arg1 Role 53:56 the Role 49:56 the Role of the Saposin Domain in Activating Substrate Hydrolysis 49:113 Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis. 27349982 7 1 theme hydrophobic 1448:1458 arg1 interactions 1460:1471 electrostatic and hydrophobic interactions 1430:1471 electrostatic and hydrophobic interactions 1430:1471 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 4 2 theme catalytic 886:894 arg1 center 896:901 the catalytic center 882:901 the catalytic center 882:901 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 2 3 theme ASM 369:371 arg1 polypeptide 373:383 the ASM polypeptide 365:383 the ASM polypeptide 365:383 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 0 4 theme Saposin 65:71 arg1 Domain 73:78 the Saposin Domain 61:78 the Saposin Domain 61:78 Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis. 27349982 3 5 theme catalytic 676:684 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 5 theme catalytic 676:684 arg1 domain 686:691 the catalytic domain 672:691 the catalytic domain 672:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 4 6 theme concave 843:849 arg1 surface 851:857 a hydrophobic concave surface 829:857 a hydrophobic concave surface approximately 30Å from the catalytic center 829:901 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 5 7 theme acidic 1069:1074 arg1 lipids 1076:1081 acidic lipids 1069:1081 acidic lipids 1069:1081 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 7 8 theme membrane-bound 1586:1599 arg1 reaction 1601:1608 a membrane-bound reaction 1584:1608 a membrane-bound reaction 1584:1608 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 5 9 with consistent 1010:1019 arg1 requirement 1037:1047 the strict requirement 1026:1047 the strict requirement for the presence of acidic lipids in target membranes 1026:1101 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 6 10 theme binuclear 1206:1214 arg1 center 1221:1226 the binuclear zinc center 1202:1226 the binuclear zinc center 1202:1226 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 6 11 theme zinc 1216:1219 arg1 center 1221:1226 the binuclear zinc center 1202:1226 the binuclear zinc center 1202:1226 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 5 12 theme lipids 1076:1081 arg1 presence 1057:1064 the presence 1053:1064 the presence of acidic lipids in target membranes 1053:1101 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 6 13 theme intrinsic 1280:1288 arg1 flexibility 1290:1300 the intrinsic flexibility 1276:1300 the intrinsic flexibility of the saposin domain 1276:1322 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 6 13 theme intrinsic 1280:1288 arg1 important 1327:1335 important 1327:1335 important 1327:1335 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 5 14 theme electrostatic 919:931 arg1 electropositive 960:974 electropositive 960:974 electropositive 960:974 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 5 14 theme electrostatic 919:931 arg1 potential 933:941 The calculated electrostatic potential 904:941 The calculated electrostatic potential of the enzyme 904:955 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 6 15 theme membrane 1372:1379 arg1 interactions 1381:1392 membrane interactions 1372:1392 membrane interactions 1372:1392 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 2 16 theme sphingolipid 286:297 arg1 hydrolases 299:308 other lysosomal sphingolipid hydrolases 270:308 other lysosomal sphingolipid hydrolases 270:308 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 5 17 theme calculated 908:917 arg1 electropositive 960:974 electropositive 960:974 electropositive 960:974 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 5 17 theme calculated 908:917 arg1 potential 933:941 The calculated electrostatic potential 904:941 The calculated electrostatic potential of the enzyme 904:955 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 1 18 theme lysosomal 149:157 arg1 sphingomyelinase 121:136 Acid sphingomyelinase 116:136 Acid sphingomyelinase (ASM) 116:142 Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. 27349982 1 18 theme lysosomal 149:157 arg1 phosphodiesterase 159:175 a lysosomal phosphodiesterase 147:175 a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine 147:261 Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. 27349982 0 19 theme Substrate 94:102 arg1 Hydrolysis 104:113 Substrate Hydrolysis 94:113 Substrate Hydrolysis 94:113 Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis. 27349982 5 20 theme target 1086:1091 arg1 membranes 1093:1101 target membranes 1086:1101 target membranes 1086:1101 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 5 21 from presence 1057:1064 arg1 membranes 1093:1101 target membranes 1086:1101 target membranes 1086:1101 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 2 22 theme lysosomal 276:284 arg1 hydrolases 299:308 other lysosomal sphingolipid hydrolases 270:308 other lysosomal sphingolipid hydrolases 270:308 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 7 23 theme local 1482:1486 arg1 disruptions 1488:1498 local disruptions 1482:1498 local disruptions of target bilayers 1482:1517 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 2 24 theme N-terminal 409:418 arg1 domain 428:433 a built-in N-terminal saposin domain 398:433 a built-in N-terminal saposin domain 398:433 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 2 25 theme activator 468:476 arg1 protein 478:484 an external activator protein 456:484 an external activator protein 456:484 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 5 26 from pH 990:991 arg1 electropositive 960:974 electropositive 960:974 electropositive 960:974 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 5 26 from pH 990:991 arg1 potential 933:941 The calculated electrostatic potential 904:941 The calculated electrostatic potential of the enzyme 904:955 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 4 27 theme hydrophobic 831:841 arg1 surface 851:857 a hydrophobic concave surface 829:857 a hydrophobic concave surface approximately 30Å from the catalytic center 829:901 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 6 28 theme saposin 1309:1315 arg1 domain 1317:1322 the saposin domain 1305:1322 the saposin domain 1305:1322 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 5 29 attach presence 1057:1064 arg2 lipids 1076:1081 acidic lipids 1069:1081 acidic lipids 1069:1081 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 5 29 attach presence 1057:1064 arg1 membranes 1093:1101 target membranes 1086:1101 target membranes 1086:1101 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 2 30 theme built-in 400:407 arg1 domain 428:433 a built-in N-terminal saposin domain 398:433 a built-in N-terminal saposin domain 398:433 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 2 31 theme external 459:466 arg1 protein 478:484 an external activator protein 456:484 an external activator protein 456:484 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 7 32 theme interactions 1460:1471 arg1 combination 1415:1425 a combination 1413:1425 a combination of electrostatic and hydrophobic interactions 1413:1471 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 2 33 theme activator 328:336 arg1 protein 338:344 a saposin activator protein 318:344 a saposin activator protein for full activity 318:362 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 7 34 theme catalytic 1564:1572 arg1 center 1574:1579 the catalytic center 1560:1579 the catalytic center in a membrane-bound reaction 1560:1608 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 0 35 theme Acid 19:22 arg1 Sphingomyelinase 24:39 Human Acid Sphingomyelinase 13:39 Human Acid Sphingomyelinase 13:39 Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis. 27349982 7 36 theme electrostatic 1430:1442 arg1 interactions 1460:1471 electrostatic and hydrophobic interactions 1430:1471 electrostatic and hydrophobic interactions 1430:1471 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 7 37 from center 1574:1579 arg1 reaction 1601:1608 a membrane-bound reaction 1584:1608 a membrane-bound reaction 1584:1608 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 3 38 dep regions 586:592 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 38 dep regions 586:592 arg1 domain 632:637 the N-terminal saposin domain 609:637 the N-terminal saposin domain 609:637 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 38 dep regions 586:592 arg1 domain 686:691 the catalytic domain 672:691 the catalytic domain 672:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 38 dep regions 586:592 arg1 connector 657:665 the proline-rich connector 640:665 the proline-rich connector 640:665 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 2 39 theme saposin 320:326 arg1 protein 338:344 a saposin activator protein 318:344 a saposin activator protein for full activity 318:362 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 6 40 theme dynamic 1242:1248 arg1 simulations 1250:1260 molecular dynamic simulations 1232:1260 molecular dynamic simulations 1232:1260 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 0 41 theme Human 13:17 arg1 Sphingomyelinase 24:39 Human Acid Sphingomyelinase 13:39 Human Acid Sphingomyelinase 13:39 Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis. 27349982 6 42 theme ceramide-phosphate 1163:1180 arg1 group 1182:1186 the ceramide-phosphate group 1159:1186 the ceramide-phosphate group positioned at the binuclear zinc center 1159:1226 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 5 43 theme enzyme 950:955 arg1 electropositive 960:974 electropositive 960:974 electropositive 960:974 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 5 43 theme enzyme 950:955 arg1 potential 933:941 The calculated electrostatic potential 904:941 The calculated electrostatic potential of the enzyme 904:955 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 3 44 theme crystal 507:513 arg1 structure 515:523 the crystal structure 503:523 the crystal structure of human ASM 503:536 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 6 45 theme monomer-dimer 1341:1353 arg1 exchange 1355:1362 monomer-dimer exchange 1341:1362 monomer-dimer exchange 1341:1362 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 3 46 theme proline-rich 644:655 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 46 theme proline-rich 644:655 arg1 connector 657:665 the proline-rich connector 640:665 the proline-rich connector 640:665 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 5 47 theme lysosome 1000:1007 arg1 pH 990:991 the acidic pH 979:991 the acidic pH of the lysosome 979:1007 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 7 48 theme lipid 1541:1545 arg1 headgroup 1547:1555 the lipid headgroup 1537:1555 the lipid headgroup 1537:1555 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 3 49 theme main 581:584 arg1 connector 657:665 the proline-rich connector 640:665 the proline-rich connector 640:665 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 49 theme main 581:584 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 49 theme main 581:584 arg1 domain 686:691 the catalytic domain 672:691 the catalytic domain 672:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 49 theme main 581:584 arg1 domain 632:637 the N-terminal saposin domain 609:637 the N-terminal saposin domain 609:637 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 0 50 theme Sphingomyelinase 24:39 arg1 Structure 0:8 Structure 0:8 Structure of Human Acid Sphingomyelinase 0:39 Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis. 27349982 7 51 theme target 1503:1508 arg1 bilayers 1510:1517 target bilayers 1503:1517 target bilayers 1503:1517 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 3 52 theme enzyme 601:606 arg1 connector 657:665 the proline-rich connector 640:665 the proline-rich connector 640:665 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 52 theme enzyme 601:606 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 52 theme enzyme 601:606 arg1 domain 686:691 the catalytic domain 672:691 the catalytic domain 672:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 52 theme enzyme 601:606 arg1 domain 632:637 the N-terminal saposin domain 609:637 the N-terminal saposin domain 609:637 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 1 53 theme sphingomyelin 210:222 arg1 hydrolysis 196:205 the hydrolysis 192:205 the hydrolysis of sphingomyelin 192:222 Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. 27349982 5 54 theme strict 1030:1035 arg1 requirement 1037:1047 the strict requirement 1026:1047 the strict requirement for the presence of acidic lipids in target membranes 1026:1101 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 3 55 theme regions 586:592 arg1 organization 555:566 the organization 551:566 the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 551:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 6 56 theme Docking 1104:1110 arg1 studies 1112:1118 Docking studies 1104:1118 Docking studies 1104:1118 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 7 57 theme bilayers 1510:1517 arg1 disruptions 1488:1498 local disruptions 1482:1498 local disruptions of target bilayers 1482:1517 Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. 27349982 2 58 theme saposin 420:426 arg1 domain 428:433 a built-in N-terminal saposin domain 398:433 a built-in N-terminal saposin domain 398:433 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 4 59 theme open 806:809 arg1 form 811:814 an open form 803:814 an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center 803:901 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 1 60 theme Acid 116:119 arg1 sphingomyelinase 121:136 Acid sphingomyelinase 116:136 Acid sphingomyelinase (ASM) 116:142 Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. 27349982 1 60 theme Acid 116:119 arg1 phosphodiesterase 159:175 a lysosomal phosphodiesterase 147:175 a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine 147:261 Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. 27349982 1 60 theme Acid 116:119 arg1 ASM 139:141 ASM 139:141 ASM 139:141 Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. 27349982 6 61 theme domain 1317:1322 arg1 flexibility 1290:1300 the intrinsic flexibility 1276:1300 the intrinsic flexibility of the saposin domain 1276:1322 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 6 61 theme domain 1317:1322 arg1 important 1327:1335 important 1327:1335 important 1327:1335 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 3 62 theme N-terminal 613:622 arg1 domain 632:637 the N-terminal saposin domain 609:637 the N-terminal saposin domain 609:637 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 62 theme N-terminal 613:622 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 2 63 theme other 270:274 arg1 hydrolases 299:308 other lysosomal sphingolipid hydrolases 270:308 other lysosomal sphingolipid hydrolases 270:308 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 6 64 theme molecular 1232:1240 arg1 simulations 1250:1260 molecular dynamic simulations 1232:1260 molecular dynamic simulations 1232:1260 Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. 27349982 2 65 theme full 350:353 arg1 activity 355:362 full activity 350:362 full activity 350:362 While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. 27349982 4 66 theme large 756:760 arg1 domain 785:790 the large, bowl-shaped catalytic domain 752:790 the large, bowl-shaped catalytic domain 752:790 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 3 67 theme saposin 624:630 arg1 domain 632:637 the N-terminal saposin domain 609:637 the N-terminal saposin domain 609:637 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 3 67 theme saposin 624:630 arg1 regions 586:592 the three main regions 571:592 the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain 571:691 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 4 68 dep large 756:760 arg1 bowl-shaped 763:773 bowl-shaped 763:773 bowl-shaped 763:773 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 4 69 theme catalytic 775:783 arg1 domain 785:790 the large, bowl-shaped catalytic domain 752:790 the large, bowl-shaped catalytic domain 752:790 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 3 70 theme human 528:532 arg1 ASM 534:536 human ASM 528:536 human ASM 528:536 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 5 71 theme acidic 983:988 arg1 pH 990:991 the acidic pH 979:991 the acidic pH of the lysosome 979:1007 The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. 27349982 4 72 theme domain 785:790 arg1 edge 744:747 an edge 741:747 an edge of the large, bowl-shaped catalytic domain 741:790 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 27349982 3 73 theme ASM 534:536 arg1 structure 515:523 the crystal structure 503:523 the crystal structure of human ASM 503:536 Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. 27349982 4 74 theme saposin 698:704 arg1 domain 706:711 The saposin domain 694:711 The saposin domain 694:711 The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. 3886654 5 0 contain containing 656:665 arg1 residues 647:654 353 amino acid residues 632:654 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains 632:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 5 0 contain containing 656:665 arg2 probable 690:697 probable 690:697 probable 690:697 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 5 0 contain containing 656:665 arg2 Asn-linked 671:680 Asn-linked 671:680 Asn-linked 671:680 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 6 1 dep " 872:872 arg1 domains 888:894 domains 888:894 domains 888:894 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 7 2 theme factor 1076:1081 arg1 XII 1083:1085 factor XII 1076:1085 factor XII 1076:1085 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 4 3 dep 258 486:488 arg1 10924-10933 491:501 10924-10933 491:501 10924-10933 491:501 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 0 4 theme XIIa 61:64 arg1 chain 33:37 the heavy chain 23:37 the heavy chain of human alpha-factor XIIa 23:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 1 5 theme intact 304:309 arg1 XII 318:320 intact factor XII and alpha-factor XIIa 304:342 XII 318:320 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 6 theme heavy 125:129 arg1 chain 131:135 the heavy chain 121:135 the heavy chain of human alpha-factor XIIa (activated Hageman factor) 121:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 7 theme automated 209:217 arg1 degradation 225:235 automated Edman degradation 209:235 automated Edman degradation 209:235 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 0 8 theme Hageman 77:83 arg1 factor 85:90 activated Hageman factor 67:90 activated Hageman factor 67:90 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 1 9 theme factor 311:316 arg1 XII 318:320 intact factor XII and alpha-factor XIIa 304:342 XII 318:320 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 2 10 dep K. 440:441 arg1 A 461:461 A 461:461 A 461:461 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 2 10 dep K. 440:441 arg1 Biol 474:477 Biol 474:477 Biol 474:477 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 1 11 theme Edman 219:223 arg1 degradation 225:235 automated Edman degradation 209:235 automated Edman degradation 209:235 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 0 12 theme activated 67:75 arg1 factor 85:90 activated Hageman factor 67:90 activated Hageman factor 67:90 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 1 13 theme XII 318:320 arg1 cleavages 291:299 chemical and enzymatic cleavages 268:299 chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa 268:342 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 7 14 theme XII 946:948 arg1 organization 923:934 The domain organization 912:934 The domain organization of factor XII 912:948 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 14 theme XII 946:948 arg1 analogous 953:961 analogous 953:961 analogous 953:961 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 6 15 contain contain 777:783 arg2 " 872:872 the "type I" 861:872 the "type I" 861:872 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 15 contain contain 777:783 arg2 domains 800:806 four different domains 785:806 four different domains including a "kringle 785:827 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 15 contain contain 777:783 arg1 chain 739:743 The heavy chain 729:743 The heavy chain of alpha-factor XIIa 729:764 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 15 contain contain 777:783 arg2 fibronectin 899:909 fibronectin 899:909 fibronectin 899:909 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 15 contain contain 777:783 arg2 kringle 821:827 a "kringle 818:827 a "kringle 818:827 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 15 contain contain 777:783 arg2 domain 849:854 " a "growth factor" domain 829:854 " a "growth factor" domain 829:854 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 7 16 theme tissue 1016:1021 arg1 activator 1035:1043 tissue plasminogen activator 1016:1043 tissue plasminogen activator 1016:1043 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 6 17 theme different 790:798 arg1 domains 800:806 four different domains 785:806 four different domains including a "kringle 785:827 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 17 theme different 790:798 arg1 fibronectin 899:909 fibronectin 899:909 fibronectin 899:909 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 17 theme different 790:798 arg1 kringle 821:827 a "kringle 818:827 a "kringle 818:827 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 5 18 theme O-linked 699:706 arg1 chains 721:726 O-linked carbohydrate chains 699:726 O-linked carbohydrate chains 699:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 1 19 theme chain 131:135 arg1 sequence 109:116 The amino acid sequence 94:116 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) 94:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 5 20 theme amino 636:640 arg1 residues 647:654 353 amino acid residues 632:654 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains 632:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 1 21 theme alpha-factor 326:337 arg1 XIIa 339:342 intact factor XII and alpha-factor XIIa 304:342 XIIa 339:342 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 6 22 theme alpha-factor 748:759 arg1 XIIa 761:764 alpha-factor XIIa 748:764 alpha-factor XIIa 748:764 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 2 23 theme beta-factor 412:422 arg1 XIIa 424:427 beta-factor XIIa 412:427 beta-factor XIIa 412:427 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 4 24 theme XII 554:556 arg1 sequence 529:536 the complete amino acid sequence 505:536 the complete amino acid sequence of human factor XII 505:556 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 4 24 theme XII 554:556 arg1 258 486:488 258 486:488 258 486:488 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 0 25 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the heavy chain of human alpha-factor XIIa 0:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 1 26 theme XIIa 339:342 arg1 cleavages 291:299 chemical and enzymatic cleavages 268:299 chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa 268:342 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 5 27 dep Asn-linked 671:680 arg1 chains 721:726 O-linked carbohydrate chains 699:726 O-linked carbohydrate chains 699:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 5 28 theme acid 642:645 arg1 residues 647:654 353 amino acid residues 632:654 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains 632:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 7 29 theme same 1102:1105 arg1 subfamily 1116:1124 the same protease subfamily 1098:1124 the same protease subfamily as these two proteins 1098:1146 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 1 30 theme human 140:144 arg1 factor 183:188 activated Hageman factor 165:188 activated Hageman factor 165:188 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 30 theme human 140:144 arg1 XIIa 159:162 human alpha-factor XIIa 140:162 human alpha-factor XIIa (activated Hageman factor) 140:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 0 31 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the heavy chain of human alpha-factor XIIa 0:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 7 32 theme domain 916:921 arg1 organization 923:934 The domain organization 912:934 The domain organization of factor XII 912:948 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 32 theme domain 916:921 arg1 analogous 953:961 analogous 953:961 analogous 953:961 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 1 33 theme alpha-factor 146:157 arg1 factor 183:188 activated Hageman factor 165:188 activated Hageman factor 165:188 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 33 theme alpha-factor 146:157 arg1 XIIa 159:162 human alpha-factor XIIa 140:162 human alpha-factor XIIa (activated Hageman factor) 140:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 7 34 theme fibrinolytic 983:994 arg1 activator 1035:1043 tissue plasminogen activator 1016:1043 tissue plasminogen activator 1016:1043 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 34 theme fibrinolytic 983:994 arg1 proteins 996:1003 several fibrinolytic proteins 975:1003 several fibrinolytic proteins 975:1003 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 34 theme fibrinolytic 983:994 arg1 urokinase 1049:1057 urokinase 1049:1057 urokinase 1049:1057 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 5 35 theme carbohydrate 708:719 arg1 chains 721:726 O-linked carbohydrate chains 699:726 O-linked carbohydrate chains 699:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 6 36 theme type 879:882 arg1 " 886:886 "type II" 878:886 "type II" 878:886 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 1 37 theme XIIa 159:162 arg1 chain 131:135 the heavy chain 121:135 the heavy chain of human alpha-factor XIIa (activated Hageman factor) 121:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 6 38 theme factor 841:846 arg1 fibronectin 899:909 fibronectin 899:909 fibronectin 899:909 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 38 theme factor 841:846 arg1 domain 849:854 " a "growth factor" domain 829:854 " a "growth factor" domain 829:854 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 5 39 theme alpha-factor 599:610 arg1 XIIa 612:615 alpha-factor XIIa 599:615 alpha-factor XIIa 599:615 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 6 40 theme II 884:885 arg1 " 886:886 "type II" 878:886 "type II" 878:886 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 0 41 theme heavy 27:31 arg1 chain 33:37 the heavy chain 23:37 the heavy chain of human alpha-factor XIIa 23:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 6 42 theme growth 834:839 arg1 factor 841:846 " a "growth factor 829:846 " a "growth factor" domain 829:854 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 5 43 theme XIIa 612:615 arg1 chain 590:594 The heavy chain 580:594 The heavy chain of alpha-factor XIIa 580:615 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 4 44 theme factor 547:552 arg1 XII 554:556 human factor XII 541:556 human factor XII 541:556 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 1 45 theme activated 165:173 arg1 factor 183:188 activated Hageman factor 165:188 activated Hageman factor 165:188 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 45 theme activated 165:173 arg1 XIIa 159:162 human alpha-factor XIIa 140:162 human alpha-factor XIIa (activated Hageman factor) 140:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 7 46 theme several 975:981 arg1 activator 1035:1043 tissue plasminogen activator 1016:1043 tissue plasminogen activator 1016:1043 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 46 theme several 975:981 arg1 proteins 996:1003 several fibrinolytic proteins 975:1003 several fibrinolytic proteins 975:1003 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 46 theme several 975:981 arg1 urokinase 1049:1057 urokinase 1049:1057 urokinase 1049:1057 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 6 47 theme XIIa 761:764 arg1 chain 739:743 The heavy chain 729:743 The heavy chain of alpha-factor XIIa 729:764 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 48 theme I 871:871 arg1 " 872:872 the "type I" 861:872 the "type I" 861:872 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 1 49 theme Hageman 175:181 arg1 factor 183:188 activated Hageman factor 165:188 activated Hageman factor 165:188 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 49 theme Hageman 175:181 arg1 XIIa 159:162 human alpha-factor XIIa 140:162 human alpha-factor XIIa (activated Hageman factor) 140:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 1 50 theme chemical 268:275 arg1 cleavages 291:299 chemical and enzymatic cleavages 268:299 chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa 268:342 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 6 51 theme " 847:847 arg1 fibronectin 899:909 fibronectin 899:909 fibronectin 899:909 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 51 theme " 847:847 arg1 domain 849:854 " a "growth factor" domain 829:854 " a "growth factor" domain 829:854 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 7 52 theme protease 1107:1114 arg1 subfamily 1116:1124 the same protease subfamily 1098:1124 the same protease subfamily as these two proteins 1098:1146 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 0 53 theme chain 33:37 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the heavy chain of human alpha-factor XIIa 0:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 7 54 theme plasminogen 1023:1033 arg1 activator 1035:1043 tissue plasminogen activator 1016:1043 tissue plasminogen activator 1016:1043 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 7 55 theme factor 939:944 arg1 XII 946:948 factor XII 939:948 factor XII 939:948 The domain organization of factor XII is analogous to those of several fibrinolytic proteins, including tissue plasminogen activator and urokinase, suggesting that factor XII belongs to the same protease subfamily as these two proteins. 3886654 1 56 theme enzymatic 281:289 arg1 cleavages 291:299 chemical and enzymatic cleavages 268:299 chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa 268:342 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 5 57 theme heavy 584:588 arg1 chain 590:594 The heavy chain 580:594 The heavy chain of alpha-factor XIIa 580:615 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 2 58 dep Combining 345:353 arg1 1983 465:468 1983 465:468 1983 465:468 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 2 58 dep Combining 345:353 arg1 B. 458:459 B. 458:459 B. 458:459 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 2 58 dep Combining 345:353 arg1 K. 440:441 K. 440:441 K. 440:441 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 1 59 theme amino 98:102 arg1 sequence 109:116 The amino acid sequence 94:116 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) 94:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 4 60 theme complete 509:516 arg1 sequence 529:536 the complete amino acid sequence 505:536 the complete amino acid sequence of human factor XII 505:556 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 4 60 theme complete 509:516 arg1 258 486:488 258 486:488 258 486:488 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 6 61 theme heavy 733:737 arg1 chain 739:743 The heavy chain 729:743 The heavy chain of alpha-factor XIIa 729:764 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 0 62 dep sequence 11:18 arg1 factor 85:90 activated Hageman factor 67:90 activated Hageman factor 67:90 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 1 63 theme acid 104:107 arg1 sequence 109:116 The amino acid sequence 94:116 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) 94:189 The amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor) was determined by automated Edman degradation using the peptides produced by chemical and enzymatic cleavages of intact factor XII and alpha-factor XIIa. 3886654 0 64 theme alpha-factor 48:59 arg1 XIIa 61:64 human alpha-factor XIIa 42:64 human alpha-factor XIIa 42:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 4 65 theme human 541:545 arg1 XII 554:556 human factor XII 541:556 human factor XII 541:556 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 5 66 link O-linked 699:706 arg1 chains 721:726 O-linked carbohydrate chains 699:726 O-linked carbohydrate chains 699:726 The heavy chain of alpha-factor XIIa is composed of 353 amino acid residues containing one Asn-linked and six probable O-linked carbohydrate chains. 3886654 2 67 theme determined 389:398 arg1 sequence 400:407 the previously determined sequence 374:407 the previously determined sequence of beta-factor XIIa 374:427 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 6 68 theme fibronectin 899:909 arg1 kringle 821:827 a "kringle 818:827 a "kringle 818:827 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 68 theme fibronectin 899:909 arg1 " 886:886 "type II" 878:886 "type II" 878:886 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 68 theme fibronectin 899:909 arg1 domain 849:854 " a "growth factor" domain 829:854 " a "growth factor" domain 829:854 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 68 theme fibronectin 899:909 arg1 domains 800:806 four different domains 785:806 four different domains including a "kringle 785:827 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 68 theme fibronectin 899:909 arg1 " 872:872 the "type I" 861:872 the "type I" 861:872 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 6 68 theme fibronectin 899:909 arg1 fibronectin 899:909 fibronectin 899:909 fibronectin 899:909 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 4 69 theme acid 524:527 arg1 sequence 529:536 the complete amino acid sequence 505:536 the complete amino acid sequence of human factor XII 505:556 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 4 69 theme acid 524:527 arg1 258 486:488 258 486:488 258 486:488 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 0 70 theme human 42:46 arg1 XIIa 61:64 human alpha-factor XIIa 42:64 human alpha-factor XIIa 42:64 Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). 3886654 6 71 theme type 866:869 arg1 " 872:872 the "type I" 861:872 the "type I" 861:872 The heavy chain of alpha-factor XIIa appears to contain four different domains including a "kringle," a "growth factor" domain, and the "type I" and "type II" domains of fibronectin. 3886654 2 72 theme XIIa 424:427 arg1 sequence 400:407 the previously determined sequence 374:407 the previously determined sequence of beta-factor XIIa 374:427 Combining this sequence with the previously determined sequence of beta-factor XIIa (Fujikawa, K., and McMullen, B. A. (1983) J. Biol. 3886654 4 73 theme amino 518:522 arg1 sequence 529:536 the complete amino acid sequence 505:536 the complete amino acid sequence of human factor XII 505:556 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3886654 4 73 theme amino 518:522 arg1 258 486:488 258 486:488 258 486:488 258, 10924-10933), the complete amino acid sequence of human factor XII has been established. 3202829 11 0 dep 5 1767:1767 arg1 J 1752:1752 J 1752:1752 J. 1752:1753 J. (1988) 249, 5. 3202829 10 1 theme Supplementary 1544:1556 arg1 SUP 1570:1572 Supplementary Publication SUP 50146 1544:1578 Supplementary Publication SUP 50146 (4 pages) 1544:1588 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 1 theme Supplementary 1544:1556 arg1 characterization 1381:1396 Detailed characterization 1372:1396 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1372:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 1 theme Supplementary 1544:1556 arg1 pages 1583:1587 4 pages 1581:1587 4 pages 1581:1587 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 2 2 theme SDS/polyacrylamide-gel 317:338 arg1 electrophoresis 340:354 SDS/polyacrylamide-gel electrophoresis 317:354 SDS/polyacrylamide-gel electrophoresis 317:354 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 7 3 theme seminal 914:920 arg1 plasma 922:927 human seminal plasma 908:927 human seminal plasma 908:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 10 4 theme Detailed 1372:1379 arg1 characterization 1381:1396 Detailed characterization 1372:1396 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1372:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 4 theme Detailed 1372:1379 arg1 SUP 1570:1572 Supplementary Publication SUP 50146 1544:1578 Supplementary Publication SUP 50146 (4 pages) 1544:1588 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 4 5 theme sugar 628:632 arg1 residues 634:641 about 45 sugar residues 619:641 about 45 sugar residues 619:641 It differs from the pancreatic enzyme in its glycosylation pattern as well, and contains about 45 sugar residues. 3202829 10 6 theme results 1418:1424 arg1 characterization 1381:1396 Detailed characterization 1372:1396 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1372:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 6 theme results 1418:1424 arg1 SUP 1570:1572 Supplementary Publication SUP 50146 1544:1578 Supplementary Publication SUP 50146 (4 pages) 1544:1588 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 7 theme British 1597:1603 arg1 Library 1605:1611 the British Library 1593:1611 the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem 1593:1749 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 3 8 from presence 467:474 arg1 C-terminus 518:527 the C-terminus 514:527 the C-terminus 514:527 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 7 9 theme threonine 1041:1049 arg1 presence 1029:1036 the presence 1025:1036 the presence of threonine at the C-terminus 1025:1067 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 3 10 theme additional 482:491 arg1 residue 503:509 an additional threonine residue 479:509 an additional threonine residue at the C-terminus 479:527 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 1 11 theme Edman 179:183 arg1 degradation 185:195 automatic Edman degradation 169:195 automatic Edman degradation 169:195 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 9 12 theme secretory 1208:1216 arg1 plasma 1264:1269 seminal plasma 1256:1269 seminal plasma 1256:1269 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 9 12 theme secretory 1208:1216 arg1 urine 1246:1250 urine 1246:1250 urine 1246:1250 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 9 12 theme secretory 1208:1216 arg1 RNases 1218:1223 the secretory RNases 1204:1223 the secretory RNases from human pancreas 1204:1243 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 7 13 theme urinary 982:988 arg1 enzyme 990:995 the major urinary enzyme 972:995 the major urinary enzyme 972:995 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 1 14 theme human 115:119 arg1 urine 121:125 human urine 115:125 human urine (RNase HUA) 115:137 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 14 theme human 115:119 arg1 HUA 134:136 RNase HUA 128:136 RNase HUA 128:136 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 6 15 gly Glycosylation 772:784 arg2 Asn-88 789:794 Asn-88 789:794 Asn-88 789:794 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 6 15 gly Glycosylation 772:784 arg1 RNases 852:857 mammalian secretory RNases 832:857 mammalian secretory RNases 832:857 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 9 16 theme seminal 1256:1262 arg1 plasma 1264:1269 seminal plasma 1256:1269 seminal plasma 1256:1269 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 9 16 theme seminal 1256:1262 arg1 RNases 1218:1223 the secretory RNases 1204:1223 the secretory RNases from human pancreas 1204:1243 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 0 17 from Differences 0:10 arg1 pattern 29:35 glycosylation pattern 15:35 glycosylation pattern of human secretory ribonucleases 15:68 Differences in glycosylation pattern of human secretory ribonucleases. 3202829 10 18 theme gel 1429:1431 arg1 filtration 1433:1442 gel filtration 1429:1442 gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1429:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 7 19 theme plasma 922:927 arg1 RNase 899:903 the major RNase 889:903 the major RNase of human seminal plasma 889:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 10 20 theme digests 1480:1486 arg1 filtration 1433:1442 gel filtration 1429:1442 gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1429:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 7 21 theme human 908:912 arg1 plasma 922:927 human seminal plasma 908:927 human seminal plasma 908:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 1 22 theme secretory 81:89 arg1 RNase 105:109 RNase 105:109 RNase 105:109 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 22 theme secretory 81:89 arg1 ribonuclease 91:102 The major secretory ribonuclease 71:102 The major secretory ribonuclease (RNase) of human urine (RNase HUA) 71:137 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 7 23 from data 881:884 arg1 RNase 899:903 the major RNase 889:903 the major RNase of human seminal plasma 889:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 9 24 theme post-translational 1306:1323 arg1 modifications 1325:1337 organ-specific post-translational modifications 1291:1337 organ-specific post-translational modifications of the one primary gene product 1291:1369 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 10 25 theme RNase 1515:1519 arg1 digests 1480:1486 tryptic and tryptic/chymotryptic digests 1447:1486 digests 1480:1486 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 5 26 theme oligosaccharide 749:763 arg1 chain 765:769 a complex-type oligosaccharide chain 734:769 a complex-type oligosaccharide chain 734:769 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 8 27 gly glycosylation 1074:1086 arg1 RNase 1113:1117 human seminal RNase 1099:1117 human seminal RNase 1099:1117 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 3 28 theme pancreatic 443:452 arg1 RNase 454:458 human pancreatic RNase 437:458 human pancreatic RNase 437:458 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 0 29 theme glycosylation 15:27 arg1 pattern 29:35 glycosylation pattern 15:35 glycosylation pattern of human secretory ribonucleases 15:68 Differences in glycosylation pattern of human secretory ribonucleases. 3202829 6 30 theme secretory 842:850 arg1 RNases 852:857 mammalian secretory RNases 832:857 mammalian secretory RNases 832:857 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 10 31 theme performic 1491:1499 arg1 RNase 1515:1519 performic acid-oxidized RNase 1491:1519 performic acid-oxidized RNase 1491:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 32 theme Yorkshire 1658:1666 arg1 7BQ 1673:1675 Yorkshire LS23 7BQ 1658:1675 Wetherby, West Yorkshire LS23 7BQ 1643:1675 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 5 33 theme complex-type 736:747 arg1 chain 765:769 a complex-type oligosaccharide chain 734:769 a complex-type oligosaccharide chain 734:769 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 0 34 theme secretory 46:54 arg1 ribonucleases 56:68 human secretory ribonucleases 40:68 human secretory ribonucleases 40:68 Differences in glycosylation pattern of human secretory ribonucleases. 3202829 9 35 theme gene 1358:1361 arg1 product 1363:1369 the one primary gene product 1342:1369 the one primary gene product 1342:1369 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 2 36 theme RNase 297:301 arg1 activities 303:312 other urine RNase activities 285:312 other urine RNase activities 285:312 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 7 37 theme major 893:897 arg1 RNase 899:903 the major RNase 889:903 the major RNase of human seminal plasma 889:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 4 38 theme glycosylation 575:587 arg1 pattern 589:595 its glycosylation pattern 571:595 its glycosylation pattern 571:595 It differs from the pancreatic enzyme in its glycosylation pattern as well, and contains about 45 sugar residues. 3202829 3 39 theme threonine 493:501 arg1 residue 503:509 an additional threonine residue 479:509 an additional threonine residue at the C-terminus 479:527 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 1 40 theme peptides 213:220 arg1 analysis 201:208 analysis 201:208 analysis 201:208 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 40 theme peptides 213:220 arg1 degradation 185:195 automatic Edman degradation 169:195 automatic Edman degradation 169:195 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 8 41 theme human 1099:1103 arg1 RNase 1113:1117 human seminal RNase 1099:1117 human seminal RNase 1099:1117 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 10 42 theme Publication 1558:1568 arg1 SUP 1570:1572 Supplementary Publication SUP 50146 1544:1578 Supplementary Publication SUP 50146 (4 pages) 1544:1588 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 42 theme Publication 1558:1568 arg1 characterization 1381:1396 Detailed characterization 1372:1396 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1372:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 42 theme Publication 1558:1568 arg1 pages 1583:1587 4 pages 1581:1587 4 pages 1581:1587 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 7 43 theme sequence 872:879 arg1 data 881:884 Preliminary sequence data 860:884 Preliminary sequence data on the major RNase of human seminal plasma 860:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 1 44 theme glycopeptides 226:238 arg1 analysis 201:208 analysis 201:208 analysis 201:208 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 44 theme glycopeptides 226:238 arg1 degradation 185:195 automatic Edman degradation 169:195 automatic Edman degradation 169:195 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 8 45 theme RNase 1113:1117 arg1 similar 1127:1133 similar 1127:1133 similar 1127:1133 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 8 45 theme RNase 1113:1117 arg1 pattern 1088:1094 The glycosylation pattern 1070:1094 The glycosylation pattern of human seminal RNase 1070:1117 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 4 46 theme pancreatic 550:559 arg1 enzyme 561:566 the pancreatic enzyme 546:566 the pancreatic enzyme in its glycosylation pattern as well 546:603 It differs from the pancreatic enzyme in its glycosylation pattern as well, and contains about 45 sugar residues. 3202829 2 47 theme other 285:289 arg1 activities 303:312 other urine RNase activities 285:312 other urine RNase activities 285:312 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 9 48 theme structural 1173:1182 arg1 differences 1184:1194 The structural differences 1169:1194 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma 1169:1269 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 5 49 theme Asn-Xaa-Ser/Thr 662:676 arg1 sequences 678:686 the three Asn-Xaa-Ser/Thr sequences 652:686 the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) 652:711 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 10 50 theme peptides 1401:1408 arg1 characterization 1381:1396 Detailed characterization 1372:1396 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1372:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 50 theme peptides 1401:1408 arg1 SUP 1570:1572 Supplementary Publication SUP 50146 1544:1578 Supplementary Publication SUP 50146 (4 pages) 1544:1588 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 1 51 theme automatic 169:177 arg1 degradation 185:195 automatic Edman degradation 169:195 automatic Edman degradation 169:195 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 8 52 theme pancreatic 1150:1159 arg1 enzyme 1161:1166 the pancreatic enzyme 1146:1166 the pancreatic enzyme 1146:1166 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 7 53 attach presence 1029:1036 arg2 threonine 1041:1049 threonine 1041:1049 threonine 1041:1049 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 7 53 attach presence 1029:1036 arg1 C-terminus 1058:1067 the C-terminus 1054:1067 the C-terminus 1054:1067 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 10 54 theme Boston 1631:1636 arg1 Division 1621:1628 Division 1621:1628 Division 1621:1628 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 54 theme Boston 1631:1636 arg1 Spa 1638:1640 Boston Spa 1631:1640 Boston Spa 1631:1640 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 6 55 located observed 809:816 arg1 RNases 852:857 mammalian secretory RNases 832:857 mammalian secretory RNases 832:857 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 6 55 located observed 809:816 arg2 Glycosylation 772:784 Glycosylation 772:784 Glycosylation at Asn-88 772:794 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 9 56 theme human 1230:1234 arg1 pancreas 1236:1243 human pancreas 1230:1243 human pancreas 1230:1243 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 2 57 theme isolated 245:252 arg1 free 277:280 free 277:280 free 277:280 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 2 57 theme isolated 245:252 arg1 enzyme 254:259 The isolated enzyme 241:259 The isolated enzyme 241:259 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 7 58 theme major 976:980 arg1 enzyme 990:995 the major urinary enzyme 972:995 the major urinary enzyme 972:995 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 1 59 theme urine 121:125 arg1 RNase 105:109 RNase 105:109 RNase 105:109 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 59 theme urine 121:125 arg1 ribonuclease 91:102 The major secretory ribonuclease 71:102 The major secretory ribonuclease (RNase) of human urine (RNase HUA) 71:137 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 60 theme RNase 128:132 arg1 urine 121:125 human urine 115:125 human urine (RNase HUA) 115:137 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 60 theme RNase 128:132 arg1 HUA 134:136 RNase HUA 128:136 RNase HUA 128:136 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 8 61 theme seminal 1105:1111 arg1 RNase 1113:1117 human seminal RNase 1099:1117 human seminal RNase 1099:1117 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 5 62 dep sequences 678:686 arg1 Asn-88 705:710 Asn-88 705:710 Asn-88 705:710 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 5 62 dep sequences 678:686 arg1 Asn-76 697:702 Asn-76 697:702 Asn-76 697:702 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 5 62 dep sequences 678:686 arg1 Asn-34 689:694 Asn-34 689:694 Asn-34 689:694 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 10 63 theme filtration 1433:1442 arg1 peptides 1401:1408 peptides 1401:1408 peptides 1401:1408 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 10 63 theme filtration 1433:1442 arg1 results 1418:1424 the results 1414:1424 the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase 1414:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 3 64 from C-terminus 518:527 arg1 presence 467:474 the presence 463:474 the presence of an additional threonine residue at the C-terminus 463:527 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 3 64 from C-terminus 518:527 arg1 residue 503:509 an additional threonine residue 479:509 an additional threonine residue at the C-terminus 479:527 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 3 65 gly glycoprotein 387:398 arg1 glycoprotein 387:398 a glycoprotein 128 amino acids long 385:419 a glycoprotein 128 amino acids long 385:419 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 4 66 from enzyme 561:566 arg1 pattern 589:595 its glycosylation pattern 571:595 its glycosylation pattern 571:595 It differs from the pancreatic enzyme in its glycosylation pattern as well, and contains about 45 sugar residues. 3202829 1 67 theme major 75:79 arg1 RNase 105:109 RNase 105:109 RNase 105:109 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 1 67 theme major 75:79 arg1 ribonuclease 91:102 The major secretory ribonuclease 71:102 The major secretory ribonuclease (RNase) of human urine (RNase HUA) 71:137 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 10 68 theme tryptic/chymotryptic 1459:1478 arg1 digests 1480:1486 tryptic and tryptic/chymotryptic digests 1447:1486 digests 1480:1486 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 5 69 gly glycosylated 716:727 arg1 sequences 678:686 the three Asn-Xaa-Ser/Thr sequences 652:686 the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) 652:711 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 5 69 gly glycosylated 716:727 arg1 Each 644:647 Each 644:647 Each 644:647 Each of the three Asn-Xaa-Ser/Thr sequences (Asn-34, Asn-76, Asn-88) is glycosylated with a complex-type oligosaccharide chain. 3202829 9 70 theme organ-specific 1291:1304 arg1 modifications 1325:1337 organ-specific post-translational modifications 1291:1337 organ-specific post-translational modifications of the one primary gene product 1291:1369 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 1 71 gly glycopeptides 226:238 arg2 glycopeptides 226:238 glycopeptides 226:238 glycopeptides 226:238 The major secretory ribonuclease (RNase) of human urine (RNase HUA) was isolated and sequenced by automatic Edman degradation and analysis of peptides and glycopeptides. 3202829 6 72 from Asn-88 789:794 arg1 Glycosylation 772:784 Glycosylation 772:784 Glycosylation at Asn-88 772:794 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 10 73 theme tryptic 1447:1453 arg1 digests 1480:1486 tryptic and tryptic/chymotryptic digests 1447:1486 digests 1480:1486 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 3 74 theme long 416:419 arg1 It 379:380 It 379:380 It 379:380 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 3 74 theme long 416:419 arg1 acids 410:414 a glycoprotein 128 amino acids long 385:419 a glycoprotein 128 amino acids long 385:419 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 7 75 from C-terminus 1058:1067 arg1 presence 1029:1036 the presence 1025:1036 the presence of threonine at the C-terminus 1025:1067 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3202829 10 76 theme acid-oxidized 1501:1513 arg1 RNase 1515:1519 performic acid-oxidized RNase 1491:1519 performic acid-oxidized RNase 1491:1519 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 4 77 contain contains 610:617 arg2 residues 634:641 about 45 sugar residues 619:641 about 45 sugar residues 619:641 It differs from the pancreatic enzyme in its glycosylation pattern as well, and contains about 45 sugar residues. 3202829 4 77 contain contains 610:617 arg1 It 530:531 It 530:531 It 530:531 It differs from the pancreatic enzyme in its glycosylation pattern as well, and contains about 45 sugar residues. 3202829 2 78 theme activity 360:367 arg1 staining 369:376 activity staining 360:376 activity staining 360:376 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 0 79 theme human 40:44 arg1 ribonucleases 56:68 human secretory ribonucleases 40:68 human secretory ribonucleases 40:68 Differences in glycosylation pattern of human secretory ribonucleases. 3202829 6 80 theme mammalian 832:840 arg1 RNases 852:857 mammalian secretory RNases 832:857 mammalian secretory RNases 832:857 Glycosylation at Asn-88 has not been observed previously in mammalian secretory RNases. 3202829 3 81 theme amino 404:408 arg1 It 379:380 It 379:380 It 379:380 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 3 81 theme amino 404:408 arg1 acids 410:414 a glycoprotein 128 amino acids long 385:419 a glycoprotein 128 amino acids long 385:419 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 10 82 dep West 1653:1656 arg1 7BQ 1673:1675 Yorkshire LS23 7BQ 1658:1675 Wetherby, West Yorkshire LS23 7BQ 1643:1675 Detailed characterization of peptides and the results of gel filtration of tryptic and tryptic/chymotryptic digests of performic acid-oxidized RNase have been deposited as Supplementary Publication SUP 50146 (4 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. 3202829 3 83 theme human 437:441 arg1 RNase 454:458 human pancreatic RNase 437:458 human pancreatic RNase 437:458 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 0 84 theme ribonucleases 56:68 arg1 pattern 29:35 glycosylation pattern 15:35 glycosylation pattern of human secretory ribonucleases 15:68 Differences in glycosylation pattern of human secretory ribonucleases. 3202829 9 85 theme primary 1350:1356 arg1 product 1363:1369 the one primary gene product 1342:1369 the one primary gene product 1342:1369 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 8 86 theme glycosylation 1074:1086 arg1 similar 1127:1133 similar 1127:1133 similar 1127:1133 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 8 86 theme glycosylation 1074:1086 arg1 pattern 1088:1094 The glycosylation pattern 1070:1094 The glycosylation pattern of human seminal RNase 1070:1117 The glycosylation pattern of human seminal RNase is very similar to that of the pancreatic enzyme. 3202829 0 87 gly glycosylation 15:27 arg1 ribonucleases 56:68 human secretory ribonucleases 40:68 human secretory ribonucleases 40:68 Differences in glycosylation pattern of human secretory ribonucleases. 3202829 3 88 theme glycoprotein 387:398 arg1 It 379:380 It 379:380 It 379:380 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 3 88 theme glycoprotein 387:398 arg1 acids 410:414 a glycoprotein 128 amino acids long 385:419 a glycoprotein 128 amino acids long 385:419 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 2 89 theme activities 303:312 arg1 free 277:280 free 277:280 free 277:280 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 2 89 theme activities 303:312 arg1 enzyme 254:259 The isolated enzyme 241:259 The isolated enzyme 241:259 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 9 90 theme product 1363:1369 arg1 modifications 1325:1337 organ-specific post-translational modifications 1291:1337 organ-specific post-translational modifications of the one primary gene product 1291:1369 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 2 91 theme urine 291:295 arg1 activities 303:312 other urine RNase activities 285:312 other urine RNase activities 285:312 The isolated enzyme was shown to be free of other urine RNase activities by SDS/polyacrylamide-gel electrophoresis and activity staining. 3202829 9 92 from pancreas 1236:1243 arg1 plasma 1264:1269 seminal plasma 1256:1269 seminal plasma 1256:1269 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 9 92 from pancreas 1236:1243 arg1 urine 1246:1250 urine 1246:1250 urine 1246:1250 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 9 92 from pancreas 1236:1243 arg1 RNases 1218:1223 the secretory RNases 1204:1223 the secretory RNases from human pancreas 1204:1243 The structural differences between the secretory RNases from human pancreas, urine and seminal plasma must originate from organ-specific post-translational modifications of the one primary gene product. 3202829 3 93 theme residue 503:509 arg1 presence 467:474 the presence 463:474 the presence of an additional threonine residue at the C-terminus 463:527 It is a glycoprotein 128 amino acids long, differing from human pancreatic RNase in the presence of an additional threonine residue at the C-terminus. 3202829 7 94 theme Preliminary 860:870 arg1 data 881:884 Preliminary sequence data 860:884 Preliminary sequence data on the major RNase of human seminal plasma 860:927 Preliminary sequence data on the major RNase of human seminal plasma have revealed no difference between it and the major urinary enzyme; their similarities include the presence of threonine at the C-terminus. 3458201 5 0 theme amino 727:731 arg1 sequence 738:745 a unique amino acid sequence 718:745 a unique amino acid sequence 718:745 alpha 1B has a unique amino acid sequence. 3458201 6 1 theme certain 871:877 arg1 light 894:898 certain immunoglobulin light and heavy chains 871:915 light 894:898 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 7 2 theme sequence 941:948 arg1 similarity 950:959 sequence similarity 941:959 sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA 941:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 2 3 dep protein 259:265 arg1 Mr 268:269 Mr 268:269 Mr approximately equal to 63,000 268:299 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 6 4 theme variable 851:858 arg1 regions 860:866 variable regions 851:866 variable regions of certain immunoglobulin light and heavy chains 851:915 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 1 5 theme function 220:227 arg1 1B-glycoprotein 171:185 alpha 1B-glycoprotein 165:185 alpha 1B-glycoprotein (alpha 1B) 165:196 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 5 theme function 220:227 arg1 protein 201:207 a protein 199:207 a protein of unknown function present in human plasma 199:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 5 6 theme acid 733:736 arg1 sequence 738:745 a unique amino acid sequence 718:745 a unique amino acid sequence 718:745 alpha 1B has a unique amino acid sequence. 3458201 7 7 theme IgM 1079:1081 arg1 transport 1058:1066 transepithelial transport 1042:1066 transepithelial transport of IgA and IgM 1042:1081 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 8 theme transepithelial 1042:1056 arg1 transport 1058:1066 transepithelial transport 1042:1066 transepithelial transport of IgA and IgM 1042:1081 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 6 9 theme alpha 776:780 arg1 1B 782:783 alpha 1B 776:783 alpha 1B 776:783 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 4 10 theme amino 669:673 arg1 acids 675:679 about 95 amino acids 660:679 about 95 amino acids 660:679 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 1 11 theme present 229:235 arg1 1B-glycoprotein 171:185 alpha 1B-glycoprotein 165:185 alpha 1B-glycoprotein (alpha 1B) 165:196 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 11 theme present 229:235 arg1 protein 201:207 a protein 199:207 a protein of unknown function present in human plasma 199:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 0 12 theme supergene 90:98 arg1 family 100:105 the immunoglobulin supergene family 71:105 the immunoglobulin supergene family 71:105 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 6 13 theme 1B 782:783 arg1 domains 765:771 several domains 757:771 several domains of alpha 1B 757:783 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 6 13 theme 1B 782:783 arg1 1B 782:783 alpha 1B 776:783 alpha 1B 776:783 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 6 13 theme 1B 782:783 arg1 third 801:805 third 801:805 third 801:805 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 6 14 theme light 894:898 arg1 regions 860:866 variable regions 851:866 variable regions of certain immunoglobulin light and heavy chains 851:915 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 0 15 theme immunoglobulin 75:88 arg1 family 100:105 the immunoglobulin supergene family 71:105 the immunoglobulin supergene family 71:105 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 5 16 contain has 714:716 arg1 1B 711:712 alpha 1B 705:712 alpha 1B 705:712 alpha 1B has a unique amino acid sequence. 3458201 5 16 contain has 714:716 arg2 sequence 738:745 a unique amino acid sequence 718:745 a unique amino acid sequence 718:745 alpha 1B has a unique amino acid sequence. 3458201 1 17 gly 1B-glycoprotein 171:185 arg1 1B-glycoprotein 171:185 alpha 1B-glycoprotein 165:185 alpha 1B-glycoprotein (alpha 1B) 165:196 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 17 gly 1B-glycoprotein 171:185 arg1 1B 194:195 alpha 1B 188:195 alpha 1B 188:195 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 17 gly 1B-glycoprotein 171:185 arg1 protein 201:207 a protein 199:207 a protein of unknown function present in human plasma 199:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 18 theme human 240:244 arg1 plasma 246:251 human plasma 240:251 human plasma 240:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 6 19 theme several 757:763 arg1 domains 765:771 several domains 757:771 several domains of alpha 1B 757:783 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 6 19 theme several 757:763 arg1 1B 782:783 alpha 1B 776:783 alpha 1B 776:783 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 6 19 theme several 757:763 arg1 third 801:805 third 801:805 third 801:805 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 8 20 theme supergene 1314:1322 arg1 family 1324:1329 the immunoglobulin supergene family 1295:1329 the immunoglobulin supergene family 1295:1329 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 8 21 theme alpha 1220:1224 arg1 1B 1226:1227 alpha 1B 1220:1227 alpha 1B 1220:1227 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 4 22 theme alpha 554:558 arg1 1B 560:561 alpha 1B 554:561 alpha 1B 554:561 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 23 theme internal 572:579 arg1 duplication 581:591 internal duplication 572:591 internal duplication 572:591 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 1 24 theme alpha 165:169 arg1 1B-glycoprotein 171:185 alpha 1B-glycoprotein 165:185 alpha 1B-glycoprotein (alpha 1B) 165:196 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 24 theme alpha 165:169 arg1 1B 194:195 alpha 1B 188:195 alpha 1B 188:195 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 24 theme alpha 165:169 arg1 protein 201:207 a protein 199:207 a protein of unknown function present in human plasma 199:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 0 25 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of human plasma alpha 0:40 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 6 26 theme significant 827:837 arg1 homology 839:846 statistically significant homology 813:846 statistically significant homology to variable regions of certain immunoglobulin light and heavy chains 813:915 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 1 27 from plasma 246:251 arg1 present 229:235 present 229:235 present 229:235 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 8 28 theme sequence 1169:1176 arg1 homology 1178:1185 its sequence homology 1165:1185 its sequence homology to immunoglobulin-like proteins 1165:1217 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 2 29 theme equal 285:289 arg1 Mr 268:269 Mr 268:269 Mr approximately equal to 63,000 268:299 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 2 30 theme N-linked 341:348 arg1 chain 335:339 a single polypeptide chain 314:339 a single polypeptide chain N-linked to four glucosamine oligosaccharides 314:385 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 0 31 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of human plasma alpha 0:40 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 4 32 theme computer 525:532 arg1 programs 534:541 several computer programs 517:541 several computer programs 517:541 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 33 theme disulfide 689:697 arg1 bond 699:702 one disulfide bond 685:702 one disulfide bond 685:702 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 7 34 theme IgA 1120:1122 arg1 receptor 1029:1036 the receptor 1025:1036 the receptor for transepithelial transport of IgA and IgM 1025:1081 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 34 theme IgA 1120:1122 arg1 component 1101:1109 the secretory component 1087:1109 the secretory component of human IgA 1087:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 4 35 theme several 517:523 arg1 programs 534:541 several computer programs 517:541 several computer programs 517:541 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 7 36 theme alpha 918:922 arg1 1B 924:925 alpha 1B 918:925 alpha 1B 918:925 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 5 37 theme alpha 705:709 arg1 1B 711:712 alpha 1B 705:712 alpha 1B 705:712 alpha 1B has a unique amino acid sequence. 3458201 1 38 theme alpha 188:192 arg1 1B-glycoprotein 171:185 alpha 1B-glycoprotein 165:185 alpha 1B-glycoprotein (alpha 1B) 165:196 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 38 theme alpha 188:192 arg1 1B 194:195 alpha 1B 188:195 alpha 1B 188:195 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 2 39 link N-linked 341:348 arg1 chain 335:339 a single polypeptide chain 314:339 a single polypeptide chain N-linked to four glucosamine oligosaccharides 314:385 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 7 40 theme human 1114:1118 arg1 IgA 1120:1122 human IgA 1114:1122 human IgA 1114:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 8 41 theme similar 1276:1282 arg1 gene 1271:1274 an ancestral gene 1258:1274 an ancestral gene similar to that of the immunoglobulin supergene family 1258:1329 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 0 42 theme plasma 29:34 arg1 alpha 36:40 human plasma alpha 23:40 human plasma alpha 23:40 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 4 43 theme structural 624:633 arg1 each 644:647 each 644:647 each 644:647 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 43 theme structural 624:633 arg1 domains 635:641 five repeating structural domains 609:641 five repeating structural domains 609:641 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 8 44 theme internal 1140:1147 arg1 duplication 1149:1159 its internal duplication 1136:1159 its internal duplication 1136:1159 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 8 45 theme ancestral 1261:1269 arg1 gene 1271:1274 an ancestral gene 1258:1274 an ancestral gene similar to that of the immunoglobulin supergene family 1258:1329 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 3 46 theme disulfide 424:432 arg1 bonds 434:438 five intrachain disulfide bonds 408:438 five intrachain disulfide bonds 408:438 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 0 47 theme human 23:27 arg1 alpha 36:40 human plasma alpha 23:40 human plasma alpha 23:40 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 4 48 theme acid 500:503 arg1 sequence 505:512 the amino acid sequence 490:512 the amino acid sequence 490:512 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 1 49 from present 229:235 arg1 plasma 246:251 human plasma 240:251 human plasma 240:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 3 50 contain contains 444:451 arg1 polypeptide 392:402 The polypeptide 388:402 The polypeptide 388:402 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 3 50 contain contains 444:451 arg2 residues 468:475 474 amino acid residues 453:475 474 amino acid residues 453:475 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 2 51 theme glucosamine 358:368 arg1 oligosaccharides 370:385 four glucosamine oligosaccharides 353:385 four glucosamine oligosaccharides 353:385 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 7 52 theme secretory 1091:1099 arg1 component 1101:1109 the secretory component 1087:1109 the secretory component of human IgA 1087:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 4 53 theme amino 494:498 arg1 sequence 505:512 the amino acid sequence 490:512 the amino acid sequence 490:512 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 7 54 theme family 1010:1015 arg1 members 970:976 other members 964:976 other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA 964:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 54 theme family 1010:1015 arg1 receptor 1029:1036 the receptor 1025:1036 the receptor for transepithelial transport of IgA and IgM 1025:1081 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 54 theme family 1010:1015 arg1 component 1101:1109 the secretory component 1087:1109 the secretory component of human IgA 1087:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 55 theme immunoglobulin 985:998 arg1 family 1010:1015 the immunoglobulin supergene family 981:1015 the immunoglobulin supergene family 981:1015 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 3 56 theme acid 463:466 arg1 residues 468:475 474 amino acid residues 453:475 474 amino acid residues 453:475 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 0 57 theme alpha 36:40 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of human plasma alpha 0:40 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 3 58 contain has 404:406 arg1 polypeptide 392:402 The polypeptide 388:402 The polypeptide 388:402 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 3 58 contain has 404:406 arg2 bonds 434:438 five intrachain disulfide bonds 408:438 five intrachain disulfide bonds 408:438 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 7 59 theme IgA 1071:1073 arg1 transport 1058:1066 transepithelial transport 1042:1066 transepithelial transport of IgA and IgM 1042:1081 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 60 theme other 964:968 arg1 members 970:976 other members 964:976 other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA 964:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 60 theme other 964:968 arg1 receptor 1029:1036 the receptor 1025:1036 the receptor for transepithelial transport of IgA and IgM 1025:1081 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 7 60 theme other 964:968 arg1 component 1101:1109 the secretory component 1087:1109 the secretory component of human IgA 1087:1122 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 1 61 theme complete 112:119 arg1 sequence 132:139 The complete amino acid sequence 108:139 The complete amino acid sequence 108:139 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 4 62 theme sequence 505:512 arg1 Analysis 478:485 Analysis 478:485 Analysis of the amino acid sequence by several computer programs 478:541 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 6 63 theme chains 910:915 arg1 regions 860:866 variable regions 851:866 variable regions of certain immunoglobulin light and heavy chains 851:915 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 4 64 contain containing 649:658 arg2 bond 699:702 one disulfide bond 685:702 one disulfide bond 685:702 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 64 contain containing 649:658 arg2 acids 675:679 about 95 amino acids 660:679 about 95 amino acids 660:679 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 64 contain containing 649:658 arg1 each 644:647 each 644:647 each 644:647 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 64 contain containing 649:658 arg1 domains 635:641 five repeating structural domains 609:641 five repeating structural domains 609:641 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 1 65 attach present 229:235 arg1 plasma 246:251 human plasma 240:251 human plasma 240:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 65 attach present 229:235 arg2 protein 201:207 a protein 199:207 a protein of unknown function present in human plasma 199:251 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 65 attach present 229:235 arg2 1B-glycoprotein 171:185 alpha 1B-glycoprotein 165:185 alpha 1B-glycoprotein (alpha 1B) 165:196 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 1 66 theme amino 121:125 arg1 sequence 132:139 The complete amino acid sequence 108:139 The complete amino acid sequence 108:139 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 8 67 theme immunoglobulin 1299:1312 arg1 family 1324:1329 the immunoglobulin supergene family 1295:1329 the immunoglobulin supergene family 1295:1329 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 7 68 theme supergene 1000:1008 arg1 family 1010:1015 the immunoglobulin supergene family 981:1015 the immunoglobulin supergene family 981:1015 alpha 1B also exhibits sequence similarity to other members of the immunoglobulin supergene family such as the receptor for transepithelial transport of IgA and IgM and the secretory component of human IgA. 3458201 8 69 theme immunoglobulin-like 1190:1208 arg1 proteins 1210:1217 immunoglobulin-like proteins 1190:1217 immunoglobulin-like proteins 1190:1217 Because of its internal duplication and its sequence homology to immunoglobulin-like proteins, alpha 1B appears to have evolved from an ancestral gene similar to that of the immunoglobulin supergene family. 3458201 1 70 theme acid 127:130 arg1 sequence 132:139 The complete amino acid sequence 108:139 The complete amino acid sequence 108:139 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 3458201 3 71 theme intrachain 413:422 arg1 bonds 434:438 five intrachain disulfide bonds 408:438 five intrachain disulfide bonds 408:438 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 4 72 theme repeating 614:622 arg1 each 644:647 each 644:647 each 644:647 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 4 72 theme repeating 614:622 arg1 domains 635:641 five repeating structural domains 609:641 five repeating structural domains 609:641 Analysis of the amino acid sequence by several computer programs shows that alpha 1B exhibits internal duplication and consists of five repeating structural domains, each containing about 95 amino acids and one disulfide bond. 3458201 2 73 theme polypeptide 323:333 arg1 chain 335:339 a single polypeptide chain 314:339 a single polypeptide chain N-linked to four glucosamine oligosaccharides 314:385 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 3 74 theme amino 457:461 arg1 residues 468:475 474 amino acid residues 453:475 474 amino acid residues 453:475 The polypeptide has five intrachain disulfide bonds and contains 474 amino acid residues. 3458201 5 75 theme unique 720:725 arg1 sequence 738:745 a unique amino acid sequence 718:745 a unique amino acid sequence 718:745 alpha 1B has a unique amino acid sequence. 3458201 6 76 theme immunoglobulin 879:892 arg1 light 894:898 certain immunoglobulin light and heavy chains 871:915 light 894:898 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 0 77 gly 1B-glycoprotein 42:56 arg1 1B-glycoprotein 42:56 1B-glycoprotein 42:56 1B-glycoprotein: homology to the immunoglobulin supergene family 42:105 Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. 3458201 2 78 theme single 316:321 arg1 chain 335:339 a single polypeptide chain 314:339 a single polypeptide chain N-linked to four glucosamine oligosaccharides 314:385 This protein (Mr approximately equal to 63,000) consists of a single polypeptide chain N-linked to four glucosamine oligosaccharides. 3458201 6 79 theme heavy 904:908 arg1 chains 910:915 certain immunoglobulin light and heavy chains 871:915 chains 910:915 However, several domains of alpha 1B, especially the third, show statistically significant homology to variable regions of certain immunoglobulin light and heavy chains. 3458201 1 80 theme unknown 212:218 arg1 function 220:227 unknown function 212:227 unknown function 212:227 The complete amino acid sequence has been determined for alpha 1B-glycoprotein (alpha 1B), a protein of unknown function present in human plasma. 22576872 4 0 theme electron 836:843 arg1 ECD 867:869 ECD 867:869 ECD 867:869 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 4 0 theme electron 836:843 arg1 dissociation 853:864 electron capture dissociation 836:864 electron capture dissociation (ECD) 836:870 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 4 1 theme variants 1012:1019 arg1 characterization 983:998 a deeper characterization 974:998 a deeper characterization of possible variants and PTMs 974:1028 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 8 2 theme magnorder 1565:1573 arg1 Boreoeutheria 1575:1587 the magnorder Boreoeutheria 1561:1587 the magnorder Boreoeutheria 1561:1587 It is therefore likely that this unusual type of glycosylation is common for (at least) species belonging to the magnorder Boreoeutheria. 22576872 4 3 theme PTMs 1025:1028 arg1 characterization 983:998 a deeper characterization 974:998 a deeper characterization of possible variants and PTMs 974:1028 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 7 4 theme glycosylation 1376:1388 arg1 Aβ 1351:1352 human Aβ 1345:1352 human Aβ (a previously unknown glycosylation type) 1345:1394 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 7 4 theme glycosylation 1376:1388 arg1 type 1390:1393 a previously unknown glycosylation type 1355:1393 a previously unknown glycosylation type 1355:1393 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 1 5 theme precursor 188:196 arg1 protein 198:204 Amyloid precursor protein 180:204 Amyloid precursor protein (APP) 180:210 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 5 theme precursor 188:196 arg1 protein 229:235 the precursor protein 215:235 the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD) 215:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 5 theme precursor 188:196 arg1 APP 207:209 APP 207:209 APP 207:209 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 3 6 theme amino 712:716 arg1 variants 723:730 amino acid variants 712:730 amino acid variants 712:730 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 5 7 theme Abnormal 1031:1038 arg1 processing 1047:1056 Abnormal APP/Aβ processing 1031:1056 Abnormal APP/Aβ processing 1031:1056 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 3 8 theme Aβ 526:527 arg1 peptides 529:536 longer Aβ peptides 519:536 longer Aβ peptides 519:536 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 9 9 theme online 1691:1696 arg1 method 1716:1721 an online top-down MS-based method 1688:1721 an online top-down MS-based method 1688:1721 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 3 10 theme post-translational 735:752 arg1 modifications 754:766 post-translational modifications 735:766 post-translational modifications (PTMs) 735:773 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 10 theme post-translational 735:752 arg1 PTMs 769:772 PTMs 769:772 PTMs 769:772 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 0 11 theme cat 155:157 arg1 fluid 173:177 cat cerebrospinal fluid 155:177 cat cerebrospinal fluid 155:177 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 3 12 theme collision-induced 552:568 arg1 spectrometry 595:606 low-energy collision-induced dissociation tandem mass spectrometry 541:606 low-energy collision-induced dissociation tandem mass spectrometry (MS/MS) 541:614 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 12 theme collision-induced 552:568 arg1 MS/MS 609:613 MS/MS 609:613 MS/MS 609:613 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 7 13 from Tyr10 1315:1319 arg1 O-glycans 1302:1310 O-glycans 1302:1310 O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), 1302:1395 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 9 14 theme detailed 1609:1616 arg1 characterization 1618:1633 a detailed characterization 1607:1633 a detailed characterization of endogenous APP/Aβ peptide species in CSF 1607:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 7 15 theme sialylated 1279:1288 arg1 core 1290:1293 sialylated core 1 1279:1295 sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), 1279:1395 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 9 16 from characterization 1618:1633 arg1 CSF 1675:1677 CSF 1675:1677 CSF 1675:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 4 17 theme APP/Aβ 936:941 arg1 species 951:957 several APP/Aβ peptide species 928:957 several APP/Aβ peptide species 928:957 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 0 18 from β 110:110 arg1 characterization 56:71 comprehensive characterization 42:71 comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein 42:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 18 from β 110:110 arg1 fragments 87:95 endogenous fragments 76:95 endogenous fragments from amyloid β and amyloid precursor protein 76:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 19 theme endogenous 76:85 arg1 fragments 87:95 endogenous fragments 76:95 endogenous fragments from amyloid β and amyloid precursor protein 76:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 2 20 theme greater 379:385 arg1 knowledge 387:395 greater knowledge 379:395 greater knowledge of different APP degradation pathways 379:433 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 9 21 theme APP/Aβ 1649:1654 arg1 species 1664:1670 endogenous APP/Aβ peptide species 1638:1670 endogenous APP/Aβ peptide species in CSF 1638:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 1 22 theme main 260:263 arg1 constituent 265:275 the main constituent 256:275 the main constituent of senile plaques in Alzheimer's disease (AD) 256:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 22 theme main 260:263 arg1 β 248:248 amyloid β 240:248 amyloid β (Aβ) 240:253 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 5 23 theme storage 1099:1105 arg1 disease 1107:1113 lysosomal storage disease 1089:1113 the lysosomal storage disease Niemann-Pick type C 1085:1133 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 4 24 theme sequence 906:913 arg1 coverage 915:922 a more comprehensive sequence coverage 885:922 a more comprehensive sequence coverage for several APP/Aβ peptide species 885:957 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 0 25 from characterization 56:71 arg1 protein 134:140 amyloid precursor protein 116:140 amyloid precursor protein 116:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 25 from characterization 56:71 arg1 β 110:110 amyloid β 102:110 amyloid β 102:110 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 26 theme amyloid 116:122 arg1 protein 134:140 amyloid precursor protein 116:140 amyloid precursor protein 116:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 2 27 theme different 400:408 arg1 pathways 426:433 different APP degradation pathways 400:433 different APP degradation pathways 400:433 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 4 28 theme possible 1003:1010 arg1 variants 1012:1019 possible variants 1003:1019 possible variants 1003:1019 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 0 29 theme nano-LC-ESI-FTICR-MS 10:29 arg1 method 31:36 An online nano-LC-ESI-FTICR-MS method 0:36 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 0:178 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 5 30 theme large 1149:1153 arg1 cat 1196:1198 cat 1196:1198 cat 1196:1198 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 5 30 theme large 1149:1153 arg1 animal 1155:1160 the major large animal 1139:1160 the major large animal used for studying this disease 1139:1191 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 9 31 from CSF 1675:1677 arg1 characterization 1618:1633 a detailed characterization 1607:1633 a detailed characterization of endogenous APP/Aβ peptide species in CSF 1607:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 9 32 theme MS-based 1707:1714 arg1 method 1716:1721 an online top-down MS-based method 1688:1721 an online top-down MS-based method 1688:1721 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 3 33 theme long 624:627 arg1 b-fragments 629:639 mainly long b-fragments 617:639 mainly long b-fragments 617:639 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 5 34 theme Niemann-Pick 1115:1126 arg1 C 1133:1133 the lysosomal storage disease Niemann-Pick type C 1085:1133 the lysosomal storage disease Niemann-Pick type C 1085:1133 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 7 35 theme cerebrospinal 1425:1437 arg1 fluid 1439:1443 cat cerebrospinal fluid 1421:1443 cat cerebrospinal fluid (CSF) 1421:1449 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 7 35 theme cerebrospinal 1425:1437 arg1 CSF 1446:1448 CSF 1446:1448 CSF 1446:1448 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 2 36 theme degradation 414:424 arg1 pathways 426:433 different APP degradation pathways 400:433 different APP degradation pathways 400:433 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 9 37 theme endogenous 1638:1647 arg1 species 1664:1670 endogenous APP/Aβ peptide species 1638:1670 endogenous APP/Aβ peptide species in CSF 1638:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 1 38 theme plaques 287:293 arg1 constituent 265:275 the main constituent 256:275 the main constituent of senile plaques in Alzheimer's disease (AD) 256:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 38 theme plaques 287:293 arg1 β 248:248 amyloid β 240:248 amyloid β (Aβ) 240:253 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 9 39 from species 1664:1670 arg1 CSF 1675:1677 CSF 1675:1677 CSF 1675:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 6 40 from substitution 1217:1228 arg1 Aβ 1251:1252 cat Aβ 1247:1252 cat Aβ 1247:1252 By ECD MS/MS, a substitution of Asp7 → Glu in cat Aβ was identified. 22576872 8 41 theme unusual 1485:1491 arg1 type 1493:1496 this unusual type 1480:1496 this unusual type of glycosylation 1480:1513 It is therefore likely that this unusual type of glycosylation is common for (at least) species belonging to the magnorder Boreoeutheria. 22576872 2 42 theme Endogenous 324:333 arg1 peptides 338:345 Endogenous Aβ peptides 324:345 Endogenous Aβ peptides 324:345 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 1 43 theme senile 280:285 arg1 plaques 287:293 senile plaques 280:293 senile plaques in Alzheimer's disease (AD) 280:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 6 44 theme Asp7 1233:1236 arg1 substitution 1217:1228 a substitution 1215:1228 a substitution of Asp7 → Glu in cat Aβ 1215:1252 By ECD MS/MS, a substitution of Asp7 → Glu in cat Aβ was identified. 22576872 7 45 theme unknown 1368:1374 arg1 Aβ 1351:1352 human Aβ 1345:1352 human Aβ (a previously unknown glycosylation type) 1345:1394 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 7 45 theme unknown 1368:1374 arg1 type 1390:1393 a previously unknown glycosylation type 1355:1393 a previously unknown glycosylation type 1355:1393 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 4 46 theme capture 845:851 arg1 ECD 867:869 ECD 867:869 ECD 867:869 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 4 46 theme capture 845:851 arg1 dissociation 853:864 electron capture dissociation 836:864 electron capture dissociation (ECD) 836:870 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 1 47 theme Amyloid 180:186 arg1 protein 198:204 Amyloid precursor protein 180:204 Amyloid precursor protein (APP) 180:210 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 47 theme Amyloid 180:186 arg1 protein 229:235 the precursor protein 215:235 the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD) 215:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 47 theme Amyloid 180:186 arg1 APP 207:209 APP 207:209 APP 207:209 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 7 48 theme human 1345:1349 arg1 Aβ 1351:1352 human Aβ 1345:1352 human Aβ (a previously unknown glycosylation type) 1345:1394 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 7 48 theme human 1345:1349 arg1 type 1390:1393 a previously unknown glycosylation type 1355:1393 a previously unknown glycosylation type 1355:1393 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 9 49 theme peptide 1656:1662 arg1 species 1664:1670 endogenous APP/Aβ peptide species 1638:1670 endogenous APP/Aβ peptide species in CSF 1638:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 6 50 theme ECD 1204:1206 arg1 MS/MS 1208:1212 ECD MS/MS 1204:1212 ECD MS/MS 1204:1212 By ECD MS/MS, a substitution of Asp7 → Glu in cat Aβ was identified. 22576872 7 51 gly sialylated 1279:1288 arg1 core 1290:1293 sialylated core 1 1279:1295 sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), 1279:1395 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 3 52 theme longer 519:524 arg1 peptides 529:536 longer Aβ peptides 519:536 longer Aβ peptides 519:536 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 53 theme acid 718:721 arg1 variants 723:730 amino acid variants 712:730 amino acid variants 712:730 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 5 54 theme APP/Aβ 1040:1045 arg1 processing 1047:1056 Abnormal APP/Aβ processing 1031:1056 Abnormal APP/Aβ processing 1031:1056 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 3 55 theme dissociation 570:581 arg1 spectrometry 595:606 low-energy collision-induced dissociation tandem mass spectrometry 541:606 low-energy collision-induced dissociation tandem mass spectrometry (MS/MS) 541:614 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 55 theme dissociation 570:581 arg1 MS/MS 609:613 MS/MS 609:613 MS/MS 609:613 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 9 56 theme species 1664:1670 arg1 characterization 1618:1633 a detailed characterization 1607:1633 a detailed characterization of endogenous APP/Aβ peptide species in CSF 1607:1677 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 6 57 theme cat 1247:1249 arg1 Aβ 1251:1252 cat Aβ 1247:1252 cat Aβ 1247:1252 By ECD MS/MS, a substitution of Asp7 → Glu in cat Aβ was identified. 22576872 3 58 theme low-energy 541:550 arg1 spectrometry 595:606 low-energy collision-induced dissociation tandem mass spectrometry 541:606 low-energy collision-induced dissociation tandem mass spectrometry (MS/MS) 541:614 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 58 theme low-energy 541:550 arg1 MS/MS 609:613 MS/MS 609:613 MS/MS 609:613 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 6 59 theme Glu 1240:1242 arg1 substitution 1217:1228 a substitution 1215:1228 a substitution of Asp7 → Glu in cat Aβ 1215:1252 By ECD MS/MS, a substitution of Asp7 → Glu in cat Aβ was identified. 22576872 8 60 dep species 1540:1546 arg1 least 1533:1537 least 1533:1537 least 1533:1537 It is therefore likely that this unusual type of glycosylation is common for (at least) species belonging to the magnorder Boreoeutheria. 22576872 3 61 theme N-terminal 786:795 arg1 half 797:800 the N-terminal half 782:800 the N-terminal half of the peptide 782:815 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 0 62 theme fragments 87:95 arg1 characterization 56:71 comprehensive characterization 42:71 comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein 42:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 3 63 theme mass 590:593 arg1 spectrometry 595:606 low-energy collision-induced dissociation tandem mass spectrometry 541:606 low-energy collision-induced dissociation tandem mass spectrometry (MS/MS) 541:614 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 63 theme mass 590:593 arg1 MS/MS 609:613 MS/MS 609:613 MS/MS 609:613 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 4 64 theme several 928:934 arg1 species 951:957 several APP/Aβ peptide species 928:957 several APP/Aβ peptide species 928:957 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 0 65 from method 31:36 arg1 human 145:149 human 145:149 human 145:149 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 65 from method 31:36 arg1 fluid 173:177 cat cerebrospinal fluid 155:177 cat cerebrospinal fluid 155:177 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 1 66 theme precursor 219:227 arg1 protein 198:204 Amyloid precursor protein 180:204 Amyloid precursor protein (APP) 180:210 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 66 theme precursor 219:227 arg1 protein 229:235 the precursor protein 215:235 the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD) 215:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 4 67 theme comprehensive 892:904 arg1 coverage 915:922 a more comprehensive sequence coverage 885:922 a more comprehensive sequence coverage for several APP/Aβ peptide species 885:957 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 0 68 theme amyloid 102:108 arg1 β 110:110 amyloid β 102:110 amyloid β 102:110 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 5 69 theme lysosomal 1089:1097 arg1 disease 1107:1113 lysosomal storage disease 1089:1113 the lysosomal storage disease Niemann-Pick type C 1085:1133 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 4 70 theme peptide 943:949 arg1 species 951:957 several APP/Aβ peptide species 928:957 several APP/Aβ peptide species 928:957 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 3 71 theme peptide 809:815 arg1 half 797:800 the N-terminal half 782:800 the N-terminal half of the peptide 782:815 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 5 72 theme disease 1107:1113 arg1 C 1133:1133 the lysosomal storage disease Niemann-Pick type C 1085:1133 the lysosomal storage disease Niemann-Pick type C 1085:1133 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 2 73 theme AD 487:488 arg1 pathology 490:498 AD pathology 487:498 AD pathology 487:498 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 0 74 theme precursor 124:132 arg1 protein 134:140 amyloid precursor protein 116:140 amyloid precursor protein 116:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 75 theme online 3:8 arg1 method 31:36 An online nano-LC-ESI-FTICR-MS method 0:36 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 0:178 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 4 76 theme deeper 976:981 arg1 characterization 983:998 a deeper characterization 974:998 a deeper characterization of possible variants and PTMs 974:1028 However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. 22576872 2 77 theme Aβ 335:336 arg1 peptides 338:345 Endogenous Aβ peptides 324:345 Endogenous Aβ peptides 324:345 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 5 78 theme major 1143:1147 arg1 cat 1196:1198 cat 1196:1198 cat 1196:1198 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 5 78 theme major 1143:1147 arg1 animal 1155:1160 the major large animal 1139:1160 the major large animal used for studying this disease 1139:1191 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 9 79 theme top-down 1698:1705 arg1 method 1716:1721 an online top-down MS-based method 1688:1721 an online top-down MS-based method 1688:1721 We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method. 22576872 0 80 theme cerebrospinal 159:171 arg1 fluid 173:177 cat cerebrospinal fluid 155:177 cat cerebrospinal fluid 155:177 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 81 theme comprehensive 42:54 arg1 characterization 56:71 comprehensive characterization 42:71 comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein 42:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 2 82 theme pathways 426:433 arg1 knowledge 387:395 greater knowledge 379:395 greater knowledge of different APP degradation pathways 379:433 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 2 83 theme APP 359:361 arg1 processing 363:372 the APP processing 355:372 the APP processing 355:372 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 5 84 theme type 1128:1131 arg1 C 1133:1133 the lysosomal storage disease Niemann-Pick type C 1085:1133 the lysosomal storage disease Niemann-Pick type C 1085:1133 Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. 22576872 2 85 theme APP 410:412 arg1 pathways 426:433 different APP degradation pathways 400:433 different APP degradation pathways 400:433 Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. 22576872 7 86 theme cat 1421:1423 arg1 fluid 1439:1443 cat cerebrospinal fluid 1421:1443 cat cerebrospinal fluid (CSF) 1421:1449 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 7 86 theme cat 1421:1423 arg1 CSF 1446:1448 CSF 1446:1448 CSF 1446:1448 Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). 22576872 3 87 theme tandem 583:588 arg1 spectrometry 595:606 low-energy collision-induced dissociation tandem mass spectrometry 541:606 low-energy collision-induced dissociation tandem mass spectrometry (MS/MS) 541:614 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 3 87 theme tandem 583:588 arg1 MS/MS 609:613 MS/MS 609:613 MS/MS 609:613 When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. 22576872 8 88 theme glycosylation 1501:1513 arg1 type 1493:1496 this unusual type 1480:1496 this unusual type of glycosylation 1480:1513 It is therefore likely that this unusual type of glycosylation is common for (at least) species belonging to the magnorder Boreoeutheria. 22576872 0 89 from protein 134:140 arg1 characterization 56:71 comprehensive characterization 42:71 comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein 42:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 0 89 from protein 134:140 arg1 fragments 87:95 endogenous fragments 76:95 endogenous fragments from amyloid β and amyloid precursor protein 76:140 An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid. 22576872 1 90 theme amyloid 240:246 arg1 constituent 265:275 the main constituent 256:275 the main constituent of senile plaques in Alzheimer's disease (AD) 256:321 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 90 theme amyloid 240:246 arg1 Aβ 251:252 Aβ 251:252 Aβ 251:252 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 22576872 1 90 theme amyloid 240:246 arg1 β 248:248 amyloid β 240:248 amyloid β (Aβ) 240:253 Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). 1710976 4 0 dep model 561:565 arg1 Biochem 581:587 Biochem 581:587 Biochem 581:587 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 4 0 dep model 561:565 arg1 Karn 568:571 Karn 568:571 Karn 568:571 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 9 1 theme acid 1019:1022 arg1 incorporation 995:1007 the transialidase-catalyzed incorporation 967:1007 the transialidase-catalyzed incorporation of sialic acid into the biantennary chain 967:1049 Band 3 originates from band 1 by the transialidase-catalyzed incorporation of sialic acid into the biantennary chain. 1710976 4 2 dep resolution 484:493 arg1 reported 511:518 reported 511:518 reported 511:518 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 11 3 theme band 1279:1282 arg1 product 1268:1274 the deamidation product 1252:1274 the deamidation product of band 1 1252:1284 Only a minor part of band 3 consists of the deamidation product of band 1. 1710976 12 4 dep residues 1461:1468 arg1 364-368 1470:1476 364-368 1470:1476 364-368 1470:1476 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 8 5 dep counterpart--with 857:873 arg1 oligosaccharide 892:906 only one neutral oligosaccharide 875:906 its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule 840:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 4 6 theme higher 477:482 arg1 resolution 484:493 markedly higher resolution 468:493 markedly higher resolution than previously reported 468:518 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 8 7 theme gene 813:816 arg1 band 791:794 band 2 791:796 band 2 791:796 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 8 7 theme gene 813:816 arg1 product 818:824 the primary gene product 801:824 the primary gene product 801:824 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 13 8 theme amylase 1592:1598 arg1 clearance 1579:1587 the clearance 1575:1587 the clearance of amylase from the systemic circulation 1575:1628 Both glycosylation and deamidation might play a role in the clearance of amylase from the systemic circulation. 1710976 13 9 theme systemic 1609:1616 arg1 circulation 1618:1628 the systemic circulation 1605:1628 the systemic circulation 1605:1628 Both glycosylation and deamidation might play a role in the clearance of amylase from the systemic circulation. 1710976 8 10 theme primary 805:811 arg1 band 791:794 band 2 791:796 band 2 791:796 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 8 10 theme primary 805:811 arg1 product 818:824 the primary gene product 801:824 the primary gene product 801:824 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 7 11 theme alternative 688:698 arg1 model 700:704 an alternative model 685:704 an alternative model that fits in with our new and previously made observations 685:763 We propose an alternative model that fits in with our new and previously made observations. 1710976 12 12 theme Peptide 1287:1293 arg1 Asn-Gly-Ser 1295:1305 Peptide Asn-Gly-Ser 1287:1305 Peptide Asn-Gly-Ser (residues 427-429) 1287:1324 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 12 12 theme Peptide 1287:1293 arg1 candidate 1347:1355 the most probable candidate 1329:1355 the most probable candidate for glycosylation 1329:1373 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 12 12 theme Peptide 1287:1293 arg1 residues 1308:1315 residues 427-429 1308:1323 residues 427-429 1308:1323 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 9 13 theme biantennary 1033:1043 arg1 chain 1045:1049 the biantennary chain 1029:1049 the biantennary chain 1029:1049 Band 3 originates from band 1 by the transialidase-catalyzed incorporation of sialic acid into the biantennary chain. 1710976 12 14 theme stretch 1432:1438 arg1 residues 1461:1468 residues 364-368 1461:1476 residues 364-368 1461:1476 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 12 14 theme stretch 1432:1438 arg1 Glu-Asn-Gly-Lys-Asp 1440:1458 the stretch Glu-Asn-Gly-Lys-Asp 1428:1458 the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) 1428:1477 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 11 15 theme minor 1219:1223 arg1 part 1225:1228 Only a minor part 1212:1228 Only a minor part of band 3 1212:1238 Only a minor part of band 3 consists of the deamidation product of band 1. 1710976 4 16 theme rapid 419:423 arg1 system 455:460 a rapid and sensitive electrophoresis system 417:460 a rapid and sensitive electrophoresis system 417:460 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 8 17 attach present 908:914 arg2 counterpart--with 857:873 its glycosylated counterpart--with 840:873 its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule 840:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 8 17 attach present 908:914 arg1 molecule 924:931 each molecule 919:931 each molecule 919:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 1 18 theme Human 109:113 arg1 alpha-amylase 124:136 Human salivary alpha-amylase 109:136 Human salivary alpha-amylase 109:136 Human salivary alpha-amylase displays multiple bands upon native polyacrylamide gel electrophoresis. 1710976 14 19 from distributions 1716:1728 arg1 disease 1744:1750 disease 1744:1750 disease 1744:1750 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 14 19 from distributions 1716:1728 arg1 health 1733:1738 health 1733:1738 health 1733:1738 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 4 20 dep Karn 568:571 arg1 al. 576:578 Karn et al. 568:578 Karn et al. 568:578 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 1 21 theme salivary 115:122 arg1 alpha-amylase 124:136 Human salivary alpha-amylase 109:136 Human salivary alpha-amylase 109:136 Human salivary alpha-amylase displays multiple bands upon native polyacrylamide gel electrophoresis. 1710976 14 22 theme amylase 1700:1706 arg1 distributions 1716:1728 amylase isozyme distributions 1700:1728 amylase isozyme distributions 1700:1728 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 0 23 theme Electrophoretic 0:14 arg1 characterization 16:31 Electrophoretic characterization 0:31 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 0:107 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 12 24 theme literature 1376:1385 arg1 data 1387:1390 literature data 1376:1390 literature data 1376:1390 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 4 25 theme development 402:412 arg1 result 388:393 a result 386:393 a result of the development of a rapid and sensitive electrophoresis system 386:460 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 6 26 theme bands 667:671 arg1 origin 645:650 the origin 641:650 the origin of the various bands 641:671 1973, 10, 341-350) is inadequate to explain the origin of the various bands. 1710976 14 27 theme produced 1817:1824 arg1 amylase 1826:1832 ectopically produced amylase 1805:1832 ectopically produced amylase 1805:1832 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 0 28 theme posttranslational 36:52 arg1 modifications 54:66 posttranslational modifications 36:66 posttranslational modifications of human parotid salivary alpha-amylase 36:106 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 11 29 theme band 1233:1236 arg1 part 1225:1228 Only a minor part 1212:1228 Only a minor part of band 3 1212:1238 Only a minor part of band 3 consists of the deamidation product of band 1. 1710976 6 30 theme various 659:665 arg1 bands 667:671 the various bands 655:671 the various bands 655:671 1973, 10, 341-350) is inadequate to explain the origin of the various bands. 1710976 3 31 theme anodal 366:371 arg1 mobility 373:380 anodal mobility 366:380 anodal mobility 366:380 The isozymes are designated 1-6, in the order of increasing anodal mobility. 1710976 10 32 with product 1152:1158 arg1 band 1203:1206 band 3 1203:1208 band 3 1203:1208 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 10 32 with product 1152:1158 arg1 oligosaccharide 1186:1200 an acidic oligosaccharide 1176:1200 an acidic oligosaccharide (band 3) 1176:1209 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 1 33 theme multiple 147:154 arg1 bands 156:160 multiple bands 147:160 multiple bands 147:160 Human salivary alpha-amylase displays multiple bands upon native polyacrylamide gel electrophoresis. 1710976 12 34 dep residues 1308:1315 arg1 427-429 1317:1323 427-429 1317:1323 427-429 1317:1323 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 11 35 theme deamidation 1256:1266 arg1 product 1268:1274 the deamidation product 1252:1274 the deamidation product of band 1 1252:1284 Only a minor part of band 3 consists of the deamidation product of band 1. 1710976 14 36 theme isozyme 1708:1714 arg1 distributions 1716:1728 amylase isozyme distributions 1700:1728 amylase isozyme distributions 1700:1728 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 10 37 theme acidic 1179:1184 arg1 band 1203:1206 band 3 1203:1208 band 3 1203:1208 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 10 37 theme acidic 1179:1184 arg1 oligosaccharide 1186:1200 an acidic oligosaccharide 1176:1200 an acidic oligosaccharide (band 3) 1176:1209 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 2 38 theme 5-6 280:282 arg1 isozymes 284:291 5-6 isozymes 280:291 5-6 isozymes 280:291 In fresh saliva, due to posttranslational modifications, a pattern of 5-6 isozymes is observed. 1710976 8 39 theme glycosylated 844:855 arg1 counterpart--with 857:873 its glycosylated counterpart--with 840:873 its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule 840:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 0 40 theme modifications 54:66 arg1 characterization 16:31 Electrophoretic characterization 0:31 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 0:107 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 4 41 theme proposed 552:559 arg1 model 561:565 a previously proposed model 539:565 a previously proposed model (Karn et al., Biochem 539:587 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 14 42 theme electrophoresis 1635:1649 arg1 tool 1682:1685 a powerful tool 1671:1685 a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase 1671:1832 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 14 42 theme electrophoresis 1635:1649 arg1 system 1651:1656 The electrophoresis system 1631:1656 The electrophoresis system described 1631:1666 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 14 43 theme alterations 1785:1795 arg1 screening 1772:1780 the screening 1768:1780 the screening of alterations seen in ectopically produced amylase 1768:1832 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 0 44 theme parotid 77:83 arg1 alpha-amylase 94:106 human parotid salivary alpha-amylase 71:106 human parotid salivary alpha-amylase 71:106 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 7 45 theme made 747:750 arg1 observations 752:763 our new and previously made observations 724:763 our new and previously made observations 724:763 We propose an alternative model that fits in with our new and previously made observations. 1710976 2 46 theme fresh 213:217 arg1 saliva 219:224 fresh saliva 213:224 fresh saliva 213:224 In fresh saliva, due to posttranslational modifications, a pattern of 5-6 isozymes is observed. 1710976 0 47 theme human 71:75 arg1 alpha-amylase 94:106 human parotid salivary alpha-amylase 71:106 human parotid salivary alpha-amylase 71:106 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 12 48 dep residues 1500:1507 arg1 474-477 1509:1515 474-477 1509:1515 474-477 1509:1515 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 4 49 theme electrophoresis 439:453 arg1 system 455:460 a rapid and sensitive electrophoresis system 417:460 a rapid and sensitive electrophoresis system 417:460 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 8 50 gly glycosylated 844:855 arg1 counterpart--with 857:873 its glycosylated counterpart--with 840:873 its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule 840:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 4 51 theme sensitive 429:437 arg1 system 455:460 a rapid and sensitive electrophoresis system 417:460 a rapid and sensitive electrophoresis system 417:460 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 8 52 theme neutral 884:890 arg1 oligosaccharide 892:906 only one neutral oligosaccharide 875:906 its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule 840:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 1 53 theme native 167:172 arg1 electrophoresis 193:207 native polyacrylamide gel electrophoresis 167:207 native polyacrylamide gel electrophoresis 167:207 Human salivary alpha-amylase displays multiple bands upon native polyacrylamide gel electrophoresis. 1710976 9 54 theme transialidase-catalyzed 971:993 arg1 incorporation 995:1007 the transialidase-catalyzed incorporation 967:1007 the transialidase-catalyzed incorporation of sialic acid into the biantennary chain 967:1049 Band 3 originates from band 1 by the transialidase-catalyzed incorporation of sialic acid into the biantennary chain. 1710976 2 55 located observed 296:303 arg1 saliva 219:224 fresh saliva 213:224 fresh saliva 213:224 In fresh saliva, due to posttranslational modifications, a pattern of 5-6 isozymes is observed. 1710976 2 55 located observed 296:303 arg2 pattern 269:275 a pattern 267:275 a pattern of 5-6 isozymes 267:291 In fresh saliva, due to posttranslational modifications, a pattern of 5-6 isozymes is observed. 1710976 6 56 dep 10 603:604 arg1 341-350 607:613 341-350 607:613 341-350 607:613 1973, 10, 341-350) is inadequate to explain the origin of the various bands. 1710976 2 57 theme posttranslational 234:250 arg1 modifications 252:264 posttranslational modifications 234:264 posttranslational modifications 234:264 In fresh saliva, due to posttranslational modifications, a pattern of 5-6 isozymes is observed. 1710976 1 58 theme polyacrylamide 174:187 arg1 electrophoresis 193:207 native polyacrylamide gel electrophoresis 167:207 native polyacrylamide gel electrophoresis 167:207 Human salivary alpha-amylase displays multiple bands upon native polyacrylamide gel electrophoresis. 1710976 10 59 theme deamidation 1140:1150 arg1 product 1152:1158 the deamidation product 1136:1158 the deamidation product of amylase with an acidic oligosaccharide (band 3) 1136:1209 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 10 59 theme deamidation 1140:1150 arg1 band 1126:1129 band 5 1126:1131 band 5 1126:1131 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 7 60 theme new 728:730 arg1 observations 752:763 our new and previously made observations 724:763 our new and previously made observations 724:763 We propose an alternative model that fits in with our new and previously made observations. 1710976 0 61 theme alpha-amylase 94:106 arg1 modifications 54:66 posttranslational modifications 36:66 posttranslational modifications of human parotid salivary alpha-amylase 36:106 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 4 62 theme system 455:460 arg1 development 402:412 the development 398:412 the development of a rapid and sensitive electrophoresis system 398:460 As a result of the development of a rapid and sensitive electrophoresis system, with markedly higher resolution than previously reported, we concluded that a previously proposed model (Karn et al., Biochem. 1710976 8 63 theme present 908:914 arg1 counterpart--with 857:873 its glycosylated counterpart--with 840:873 its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule 840:931 According to this model, band 2 is the primary gene product and band 1 is its glycosylated counterpart--with only one neutral oligosaccharide present on each molecule. 1710976 1 64 theme gel 189:191 arg1 electrophoresis 193:207 native polyacrylamide gel electrophoresis 167:207 native polyacrylamide gel electrophoresis 167:207 Human salivary alpha-amylase displays multiple bands upon native polyacrylamide gel electrophoresis. 1710976 10 65 theme amylase 1163:1169 arg1 product 1152:1158 the deamidation product 1136:1158 the deamidation product of amylase with an acidic oligosaccharide (band 3) 1136:1209 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 10 65 theme amylase 1163:1169 arg1 band 1126:1129 band 5 1126:1131 band 5 1126:1131 Bands 4 and 6 originate from bands 2 and 4, respectively, by deamidation; band 5 is the deamidation product of amylase with an acidic oligosaccharide (band 3). 1710976 0 66 theme salivary 85:92 arg1 alpha-amylase 94:106 human parotid salivary alpha-amylase 71:106 human parotid salivary alpha-amylase 71:106 Electrophoretic characterization of posttranslational modifications of human parotid salivary alpha-amylase. 1710976 12 67 theme probable 1338:1345 arg1 Asn-Gly-Ser 1295:1305 Peptide Asn-Gly-Ser 1287:1305 Peptide Asn-Gly-Ser (residues 427-429) 1287:1324 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 12 67 theme probable 1338:1345 arg1 candidate 1347:1355 the most probable candidate 1329:1355 the most probable candidate for glycosylation 1329:1373 Peptide Asn-Gly-Ser (residues 427-429) is the most probable candidate for glycosylation; literature data suggests that deamidation occurs in the stretch Glu-Asn-Gly-Lys-Asp (residues 364-368) and Asn-Gly-Asn-Cys (residues 474-477). 1710976 2 68 theme isozymes 284:291 arg1 pattern 269:275 a pattern 267:275 a pattern of 5-6 isozymes 267:291 In fresh saliva, due to posttranslational modifications, a pattern of 5-6 isozymes is observed. 1710976 14 69 theme powerful 1673:1680 arg1 tool 1682:1685 a powerful tool 1671:1685 a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase 1671:1832 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 14 69 theme powerful 1673:1680 arg1 system 1651:1656 The electrophoresis system 1631:1656 The electrophoresis system described 1631:1666 The electrophoresis system described is a powerful tool to determine amylase isozyme distributions in health and disease, especially for the screening of alterations seen in ectopically produced amylase. 1710976 9 70 theme sialic 1012:1017 arg1 acid 1019:1022 sialic acid 1012:1022 sialic acid 1012:1022 Band 3 originates from band 1 by the transialidase-catalyzed incorporation of sialic acid into the biantennary chain. 1710976 13 71 from circulation 1618:1628 arg1 clearance 1579:1587 the clearance 1575:1587 the clearance of amylase from the systemic circulation 1575:1628 Both glycosylation and deamidation might play a role in the clearance of amylase from the systemic circulation. 17215869 0 0 theme H-ficolins 79:88 arg1 specificities 55:67 the innate immune recognition specificities 25:67 the innate immune recognition specificities of L- and H-ficolins 25:88 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 17215869 1 1 theme few 147:149 arg1 molecules 171:179 a few pattern recognition molecules 145:179 a few pattern recognition molecules 145:179 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 7 2 theme additional 1012:1021 arg1 S2-S4 1038:1042 S2-S4 1038:1042 S2-S4 1038:1042 In L-ficolin, three additional binding sites (S2-S4) surround the cleft. 17215869 7 2 theme additional 1012:1021 arg1 sites 1031:1035 three additional binding sites 1006:1035 three additional binding sites (S2-S4) 1006:1043 In L-ficolin, three additional binding sites (S2-S4) surround the cleft. 17215869 7 3 theme binding 1023:1029 arg1 S2-S4 1038:1042 S2-S4 1038:1042 S2-S4 1038:1042 In L-ficolin, three additional binding sites (S2-S4) surround the cleft. 17215869 7 3 theme binding 1023:1029 arg1 sites 1031:1035 three additional binding sites 1006:1035 three additional binding sites (S2-S4) 1006:1043 In L-ficolin, three additional binding sites (S2-S4) surround the cleft. 17215869 8 4 theme unpredicted 1090:1100 arg1 surface 1125:1131 an unpredicted continuous recognition surface 1087:1131 an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers 1087:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 2 5 theme defence 335:341 arg1 proteins 343:350 soluble oligomeric defence proteins 316:350 soluble oligomeric defence proteins 316:350 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 5 theme defence 335:341 arg1 H-ficolins 301:310 H-ficolins 301:310 H-ficolins 301:310 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 5 theme defence 335:341 arg1 L- 294:295 Human L- 288:295 Human L- 288:295 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 8 6 theme acetylated 1155:1164 arg1 markers 1191:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 3 7 theme various 546:552 arg1 ligands 554:560 various ligands 546:560 various ligands 546:560 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 6 8 theme binding 848:854 arg1 S1 863:864 S1 863:864 S1 863:864 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 8 theme binding 848:854 arg1 sites 856:860 Outer binding sites 842:860 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A 842:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 3 9 with complex 533:539 arg1 ligands 554:560 various ligands 546:560 various ligands 546:560 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 8 10 theme microbial 1281:1289 arg1 surfaces 1304:1311 microbial or apoptotic surfaces 1281:1311 microbial or apoptotic surfaces 1281:1311 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 5 11 theme Ca 730:731 arg1 ions 737:740 Ca(2+) ions 730:740 Ca(2+) ions 730:740 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 3 12 theme recognition 499:509 arg1 domains 511:517 their trimeric recognition domains 484:517 their trimeric recognition domains 484:517 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 8 13 theme continuous 1102:1111 arg1 surface 1125:1131 an unpredicted continuous recognition surface 1087:1131 an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers 1087:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 8 14 theme polysaccharides 1226:1240 arg1 context 1206:1212 the context 1202:1212 the context of extended polysaccharides such as 1,3-beta-D-glucan 1202:1266 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 6 15 attach present 919:925 arg1 ficolins 934:941 the ficolins 930:941 the ficolins 930:941 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 15 attach present 919:925 arg2 sites 856:860 Outer binding sites 842:860 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A 842:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 15 attach present 919:925 arg2 S1 863:864 S1 863:864 S1 863:864 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 1 16 theme recognition 159:169 arg1 molecules 171:179 a few pattern recognition molecules 145:179 a few pattern recognition molecules 145:179 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 5 17 theme homologous 761:770 arg1 sites 755:759 sites 755:759 sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin 755:839 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 1 18 from level 281:285 arg1 interactions 254:265 these interactions 248:265 these interactions at the atomic level 248:285 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 8 19 theme recognition 1113:1123 arg1 surface 1125:1131 an unpredicted continuous recognition surface 1087:1131 an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers 1087:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 1 20 theme molecules 171:179 arg1 ability 134:140 the ability 130:140 the ability of a few pattern recognition molecules to sense molecular markers on pathogens 130:219 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 6 21 from ficolins 934:941 arg1 present 919:925 present 919:925 present 919:925 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 5 22 located found 746:750 arg1 sites 755:759 sites 755:759 sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin 755:839 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 5 22 located found 746:750 arg2 ions 737:740 Ca(2+) ions 730:740 Ca(2+) ions 730:740 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 6 23 theme Outer 842:846 arg1 S1 863:864 S1 863:864 S1 863:864 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 23 theme Outer 842:846 arg1 sites 856:860 Outer binding sites 842:860 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A 842:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 0 24 theme Structural 0:9 arg1 insights 11:18 Structural insights 0:18 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 0:89 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 17215869 4 25 contain have 644:647 arg2 structures 661:670 three-lobed structures 649:670 three-lobed structures 649:670 Both domains have three-lobed structures with clefts separating the distal parts of the protomers. 17215869 4 25 contain have 644:647 arg1 domains 636:642 Both domains 631:642 Both domains 631:642 Both domains have three-lobed structures with clefts separating the distal parts of the protomers. 17215869 1 26 theme Innate 91:96 arg1 immunity 98:105 Innate immunity 91:105 Innate immunity 91:105 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 1 27 theme sense 184:188 arg1 markers 200:206 sense molecular markers 184:206 sense molecular markers on pathogens 184:219 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 2 28 theme recognition 439:449 arg1 domains 451:457 fibrinogen-like recognition domains 423:457 fibrinogen-like recognition domains 423:457 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 4 29 theme protomers 719:727 arg1 parts 706:710 the distal parts 695:710 the distal parts of the protomers 695:727 Both domains have three-lobed structures with clefts separating the distal parts of the protomers. 17215869 2 30 theme Human 288:292 arg1 proteins 343:350 soluble oligomeric defence proteins 316:350 soluble oligomeric defence proteins 316:350 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 30 theme Human 288:292 arg1 H-ficolins 301:310 H-ficolins 301:310 H-ficolins 301:310 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 30 theme Human 288:292 arg1 L- 294:295 Human L- 288:295 Human L- 288:295 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 1 31 theme molecular 190:198 arg1 markers 200:206 sense molecular markers 184:206 sense molecular markers on pathogens 184:219 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 2 32 theme fibrinogen-like 423:437 arg1 domains 451:457 fibrinogen-like recognition domains 423:457 fibrinogen-like recognition domains 423:457 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 1 33 theme atomic 274:279 arg1 level 281:285 the atomic level 270:285 the atomic level 270:285 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 6 34 located present 919:925 arg1 ficolins 934:941 the ficolins 930:941 the ficolins 930:941 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 34 located present 919:925 arg2 sites 856:860 Outer binding sites 842:860 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A 842:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 34 located present 919:925 arg2 S1 863:864 S1 863:864 S1 863:864 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 1 35 theme pattern 151:157 arg1 molecules 171:179 a few pattern recognition molecules 145:179 a few pattern recognition molecules 145:179 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 0 36 theme immune 36:41 arg1 specificities 55:67 the innate immune recognition specificities 25:67 the innate immune recognition specificities of L- and H-ficolins 25:88 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 17215869 3 37 theme domains 511:517 arg1 structures 470:479 The X-ray structures 460:479 The X-ray structures 460:479 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 4 38 theme three-lobed 649:659 arg1 structures 661:670 three-lobed structures 649:670 three-lobed structures 649:670 Both domains have three-lobed structures with clefts separating the distal parts of the protomers. 17215869 8 39 theme extended 1217:1224 arg1 polysaccharides 1226:1240 extended polysaccharides 1217:1240 extended polysaccharides such as 1,3-beta-D-glucan 1217:1266 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 8 39 theme extended 1217:1224 arg1 1,3-beta-D-glucan 1250:1266 1,3-beta-D-glucan 1250:1266 1,3-beta-D-glucan 1250:1266 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 0 40 theme innate 29:34 arg1 specificities 55:67 the innate immune recognition specificities 25:67 the innate immune recognition specificities of L- and H-ficolins 25:88 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 17215869 6 41 theme GlcNAc-binding 885:898 arg1 pocket 900:905 the GlcNAc-binding pocket 881:905 the GlcNAc-binding pocket of TL5A 881:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 8 42 theme sense 1141:1145 arg1 markers 1191:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 2 43 theme collagen 394:401 arg1 fibers 403:408 collagen fibers 394:408 collagen fibers prolonged by fibrinogen-like recognition domains 394:457 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 8 44 theme various 1147:1153 arg1 markers 1191:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 6 45 theme TL5A 910:913 arg1 pocket 900:905 the GlcNAc-binding pocket 881:905 the GlcNAc-binding pocket of TL5A 881:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 2 46 theme oligomeric 324:333 arg1 proteins 343:350 soluble oligomeric defence proteins 316:350 soluble oligomeric defence proteins 316:350 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 46 theme oligomeric 324:333 arg1 H-ficolins 301:310 H-ficolins 301:310 H-ficolins 301:310 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 46 theme oligomeric 324:333 arg1 L- 294:295 Human L- 288:295 Human L- 288:295 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 0 47 theme recognition 43:53 arg1 specificities 55:67 the innate immune recognition specificities 25:67 the innate immune recognition specificities of L- and H-ficolins 25:88 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 17215869 2 48 with proteins 343:350 arg1 activity 369:376 lectin-like activity 357:376 lectin-like activity 357:376 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 6 49 from present 919:925 arg1 ficolins 934:941 the ficolins 930:941 the ficolins 930:941 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 2 50 theme soluble 316:322 arg1 proteins 343:350 soluble oligomeric defence proteins 316:350 soluble oligomeric defence proteins 316:350 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 50 theme soluble 316:322 arg1 H-ficolins 301:310 H-ficolins 301:310 H-ficolins 301:310 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 2 50 theme soluble 316:322 arg1 L- 294:295 Human L- 288:295 Human L- 288:295 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 6 51 theme homologous 867:876 arg1 S1 863:864 S1 863:864 S1 863:864 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 6 51 theme homologous 867:876 arg1 sites 856:860 Outer binding sites 842:860 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A 842:913 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 1 52 from markers 200:206 arg1 pathogens 211:219 pathogens 211:219 pathogens 211:219 Innate immunity relies critically upon the ability of a few pattern recognition molecules to sense molecular markers on pathogens, but little is known about these interactions at the atomic level. 17215869 5 53 theme invertebrate 821:832 arg1 lectin 834:839 an invertebrate lectin 818:839 an invertebrate lectin 818:839 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 5 53 theme invertebrate 821:832 arg1 5A 807:808 tachylectin 5A 795:808 tachylectin 5A (TL5A) 795:815 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 8 54 theme apoptotic 1294:1302 arg1 surfaces 1304:1311 microbial or apoptotic surfaces 1281:1311 microbial or apoptotic surfaces 1281:1311 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 3 55 dep A 614:614 arg1 up 595:596 up 595:596 up 595:596 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 3 56 theme trimeric 490:497 arg1 domains 511:517 their trimeric recognition domains 484:517 their trimeric recognition domains 484:517 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 6 57 theme different 952:960 arg1 structures 962:971 different structures 952:971 different structures 952:971 Outer binding sites (S1) homologous to the GlcNAc-binding pocket of TL5A are present in the ficolins but show different structures and specificities. 17215869 0 58 theme L- 72:73 arg1 specificities 55:67 the innate immune recognition specificities 25:67 the innate immune recognition specificities of L- and H-ficolins 25:88 Structural insights into the innate immune recognition specificities of L- and H-ficolins. 17215869 3 59 theme X-ray 464:468 arg1 structures 470:479 The X-ray structures 460:479 The X-ray structures 460:479 The X-ray structures of their trimeric recognition domains, alone and in complex with various ligands, have been solved to resolutions up to 1.95 and 1.7 A, respectively. 17215869 5 60 theme tachylectin 795:805 arg1 lectin 834:839 an invertebrate lectin 818:839 an invertebrate lectin 818:839 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 5 60 theme tachylectin 795:805 arg1 TL5A 811:814 TL5A 811:814 TL5A 811:814 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 5 60 theme tachylectin 795:805 arg1 5A 807:808 tachylectin 5A 795:808 tachylectin 5A (TL5A) 795:815 Ca(2+) ions are found at sites homologous to those described for tachylectin 5A (TL5A), an invertebrate lectin. 17215869 2 61 theme lectin-like 357:367 arg1 activity 369:376 lectin-like activity 357:376 lectin-like activity 357:376 Human L- and H-ficolins are soluble oligomeric defence proteins with lectin-like activity, assembled from collagen fibers prolonged by fibrinogen-like recognition domains. 17215869 8 62 theme neutral 1170:1176 arg1 markers 1191:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 8 63 theme able 1133:1136 arg1 surface 1125:1131 an unpredicted continuous recognition surface 1087:1131 an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers 1087:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 8 64 theme carbohydrate 1178:1189 arg1 markers 1191:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 sense various acetylated and neutral carbohydrate markers 1141:1197 Together, they define an unpredicted continuous recognition surface able to sense various acetylated and neutral carbohydrate markers in the context of extended polysaccharides such as 1,3-beta-D-glucan, as found on microbial or apoptotic surfaces. 17215869 4 65 theme distal 699:704 arg1 parts 706:710 the distal parts 695:710 the distal parts of the protomers 695:727 Both domains have three-lobed structures with clefts separating the distal parts of the protomers. 21167756 5 0 theme tryptophan-fluorescence 740:762 arg1 quenching 764:772 tryptophan-fluorescence quenching 740:772 tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen 740:824 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 4 1 theme classical 658:666 arg1 molecules 701:709 classical major histocompatibility complex molecules 658:709 classical major histocompatibility complex molecules 658:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 2 theme pocket 536:541 arg1 architecture 543:554 an open F' pocket architecture 525:554 an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules 525:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 2 theme pocket 536:541 arg1 reminiscent 616:626 reminiscent 616:626 reminiscent of peptide binding grooves of classical major histocompatibility complex molecules 616:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 2 theme pocket 536:541 arg1 striking 512:519 striking 512:519 striking 512:519 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 0 3 theme mycobacterial 46:58 arg1 lipid 60:64 a mycobacterial lipid 44:64 a mycobacterial lipid 44:64 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 5 4 theme peptide 883:889 arg1 moieties 891:898 both the lipid and peptide moieties 864:898 moieties 891:898 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 1 5 theme CD1 138:140 arg1 molecules 142:150 CD1 molecules 138:150 CD1 molecules 138:150 CD1 molecules function to present lipid-based antigens to T cells. 21167756 2 6 attach present 213:219 arg1 complex 275:281 complex 275:281 complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 275:372 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 2 6 attach present 213:219 arg2 we 210:211 we 210:211 we 210:211 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 5 7 theme lipopeptides 956:967 arg1 presentation 940:951 CD1c presentation 935:951 CD1c presentation of lipopeptides 935:967 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 2 8 theme 2.5 Å 254:258 arg1 resolution 260:269 2.5 Å resolution 254:269 2.5 Å resolution 254:269 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 2 9 with complex 275:281 arg1 tuberculosis 317:328 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 3 10 theme unique 463:468 arg1 portal 475:480 a unique exit portal 461:480 a unique exit portal underneath the α1 helix 461:504 CD1c accommodated MPM's methylated alkyl chain exclusively in the A' pocket, aided by a unique exit portal underneath the α1 helix. 21167756 5 11 theme lipopeptide 907:917 arg1 lipid 873:877 both the lipid and peptide moieties 864:898 lipid 873:877 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 5 11 theme lipopeptide 907:917 arg1 moieties 891:898 both the lipid and peptide moieties 864:898 moieties 891:898 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 0 12 theme open 77:80 arg1 groove 82:87 an open groove 74:87 an open groove ideally suited for diverse antigen presentation 74:135 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 3 13 theme alkyl 410:414 arg1 chain 416:420 MPM's methylated alkyl chain 393:420 MPM's methylated alkyl chain 393:420 CD1c accommodated MPM's methylated alkyl chain exclusively in the A' pocket, aided by a unique exit portal underneath the α1 helix. 21167756 2 14 theme CD1c 246:249 arg1 structure 233:241 the crystal structure 221:241 the crystal structure of CD1c at 2.5 Å resolution 221:269 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 5 15 theme dodecameric 794:804 arg1 antigen 818:824 a dodecameric lipopeptide antigen 792:824 a dodecameric lipopeptide antigen 792:824 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 4 16 theme peptide 631:637 arg1 grooves 647:653 peptide binding grooves 631:653 peptide binding grooves of classical major histocompatibility complex molecules 631:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 5 17 theme lipopeptide 806:816 arg1 antigen 818:824 a dodecameric lipopeptide antigen 792:824 a dodecameric lipopeptide antigen 792:824 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 3 18 dep chain 416:420 arg1 pocket 444:449 the A' pocket 437:449 the A' pocket 437:449 CD1c accommodated MPM's methylated alkyl chain exclusively in the A' pocket, aided by a unique exit portal underneath the α1 helix. 21167756 5 19 theme antigen 818:824 arg1 loading 781:787 loading 781:787 loading of a dodecameric lipopeptide antigen 781:824 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 5 20 theme CD1c 935:938 arg1 presentation 940:951 CD1c presentation 935:951 CD1c presentation of lipopeptides 935:967 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 1 21 theme lipid-based 172:182 arg1 antigens 184:191 lipid-based antigens 172:191 lipid-based antigens 172:191 CD1 molecules function to present lipid-based antigens to T cells. 21167756 0 22 theme Å 8:8 arg1 structure 10:18 The 2.5 Å structure 0:18 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid 0:64 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 4 23 theme F 533:533 arg1 architecture 543:554 an open F' pocket architecture 525:554 an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules 525:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 23 theme F 533:533 arg1 reminiscent 616:626 reminiscent 616:626 reminiscent of peptide binding grooves of classical major histocompatibility complex molecules 616:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 23 theme F 533:533 arg1 striking 512:519 striking 512:519 striking 512:519 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 2 24 dep tuberculosis 317:328 arg1 MPM 369:371 MPM 369:371 MPM 369:371 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 2 24 dep tuberculosis 317:328 arg1 mannosyl-β1-phosphomycoketide 338:366 antigen mannosyl-β1-phosphomycoketide 330:366 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 0 25 theme CD1c 23:26 arg1 structure 10:18 The 2.5 Å structure 0:18 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid 0:64 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 4 26 theme open 528:531 arg1 architecture 543:554 an open F' pocket architecture 525:554 an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules 525:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 26 theme open 528:531 arg1 reminiscent 616:626 reminiscent 616:626 reminiscent of peptide binding grooves of classical major histocompatibility complex molecules 616:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 26 theme open 528:531 arg1 striking 512:519 striking 512:519 striking 512:519 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 0 27 theme diverse 108:114 arg1 presentation 124:135 diverse antigen presentation 108:135 diverse antigen presentation 108:135 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 5 28 theme compelling 838:847 arg1 model 849:853 a compelling model 836:853 a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides 836:967 This feature, combined with tryptophan-fluorescence quenching during loading of a dodecameric lipopeptide antigen, provides a compelling model by which both the lipid and peptide moieties of the lipopeptide are involved in CD1c presentation of lipopeptides. 21167756 4 29 theme molecules 605:613 arg1 structure 582:590 the closed cavity structure 564:590 the closed cavity structure of other CD1 molecules 564:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 30 theme major 668:672 arg1 molecules 701:709 classical major histocompatibility complex molecules 658:709 classical major histocompatibility complex molecules 658:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 31 theme CD1 601:603 arg1 molecules 605:613 other CD1 molecules 595:613 other CD1 molecules 595:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 2 32 theme antigen 330:336 arg1 MPM 369:371 MPM 369:371 MPM 369:371 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 2 32 theme antigen 330:336 arg1 mannosyl-β1-phosphomycoketide 338:366 antigen mannosyl-β1-phosphomycoketide 330:366 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 3 33 theme methylated 399:408 arg1 chain 416:420 MPM's methylated alkyl chain 393:420 MPM's methylated alkyl chain 393:420 CD1c accommodated MPM's methylated alkyl chain exclusively in the A' pocket, aided by a unique exit portal underneath the α1 helix. 21167756 0 34 theme antigen 116:122 arg1 presentation 124:135 diverse antigen presentation 108:135 diverse antigen presentation 108:135 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 3 35 theme exit 470:473 arg1 portal 475:480 a unique exit portal 461:480 a unique exit portal underneath the α1 helix 461:504 CD1c accommodated MPM's methylated alkyl chain exclusively in the A' pocket, aided by a unique exit portal underneath the α1 helix. 21167756 4 36 theme histocompatibility 674:691 arg1 molecules 701:709 classical major histocompatibility complex molecules 658:709 classical major histocompatibility complex molecules 658:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 37 theme binding 639:645 arg1 grooves 647:653 peptide binding grooves 631:653 peptide binding grooves of classical major histocompatibility complex molecules 631:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 2 38 theme Mycobacterium 303:315 arg1 tuberculosis 317:328 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 4 39 theme closed 568:573 arg1 structure 582:590 the closed cavity structure 564:590 the closed cavity structure of other CD1 molecules 564:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 40 theme molecules 701:709 arg1 grooves 647:653 peptide binding grooves 631:653 peptide binding grooves of classical major histocompatibility complex molecules 631:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 0 41 with complex 31:37 arg1 lipid 60:64 a mycobacterial lipid 44:64 a mycobacterial lipid 44:64 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 2 42 theme the pathogenic 288:301 arg1 tuberculosis 317:328 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM) 288:372 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 2 43 from resolution 260:269 arg1 structure 233:241 the crystal structure 221:241 the crystal structure of CD1c at 2.5 Å resolution 221:269 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 4 44 theme complex 693:699 arg1 molecules 701:709 classical major histocompatibility complex molecules 658:709 classical major histocompatibility complex molecules 658:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 2 45 theme crystal 225:231 arg1 structure 233:241 the crystal structure 221:241 the crystal structure of CD1c at 2.5 Å resolution 221:269 Here we present the crystal structure of CD1c at 2.5 Å resolution, in complex with the pathogenic Mycobacterium tuberculosis antigen mannosyl-β1-phosphomycoketide (MPM). 21167756 4 46 theme other 595:599 arg1 molecules 605:613 other CD1 molecules 595:613 other CD1 molecules 595:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 47 theme grooves 647:653 arg1 architecture 543:554 an open F' pocket architecture 525:554 an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules 525:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 4 47 theme grooves 647:653 arg1 reminiscent 616:626 reminiscent 616:626 reminiscent of peptide binding grooves of classical major histocompatibility complex molecules 616:709 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 21167756 0 48 from structure 10:18 arg1 complex 31:37 complex 31:37 complex with a mycobacterial lipid 31:64 The 2.5 Å structure of CD1c in complex with a mycobacterial lipid reveals an open groove ideally suited for diverse antigen presentation. 21167756 3 49 theme α1 497:498 arg1 helix 500:504 the α1 helix 493:504 the α1 helix 493:504 CD1c accommodated MPM's methylated alkyl chain exclusively in the A' pocket, aided by a unique exit portal underneath the α1 helix. 21167756 4 50 theme cavity 575:580 arg1 structure 582:590 the closed cavity structure 564:590 the closed cavity structure of other CD1 molecules 564:613 Most striking was an open F' pocket architecture lacking the closed cavity structure of other CD1 molecules, reminiscent of peptide binding grooves of classical major histocompatibility complex molecules. 16815919 7 0 theme force-field 1104:1114 arg1 TCPEp 1116:1120 the flexible and polarizable force-field TCPEp 1075:1120 the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) 1075:1182 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 5 1 theme alkaline 815:822 arg1 isozyme 854:860 a related isozyme 844:860 a related isozyme of TNAP 844:868 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 1 theme alkaline 815:822 arg1 PLAP 837:840 PLAP 837:840 PLAP 837:840 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 1 theme alkaline 815:822 arg1 phosphatase 824:834 human placental alkaline phosphatase 799:834 strontium-substituted human placental alkaline phosphatase (PLAP) 777:841 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 2 theme TNAP 865:868 arg1 isozyme 854:860 a related isozyme 844:860 a related isozyme of TNAP 844:868 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 2 theme TNAP 865:868 arg1 phosphatase 824:834 human placental alkaline phosphatase 799:834 strontium-substituted human placental alkaline phosphatase (PLAP) 777:841 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 10 3 theme doses 1644:1648 arg1 ingestion 1626:1634 ingestion 1626:1634 ingestion of high doses of strontium 1626:1661 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 0 4 from studies 11:17 arg1 complex 52:58 complex 52:58 complex with strontium 52:73 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 7 5 theme TCPEp 1116:1120 arg1 use 1068:1070 The use 1064:1070 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) 1064:1182 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 9 6 theme Energy 1446:1451 arg1 calculations 1453:1464 Energy calculations 1446:1464 Energy calculations 1446:1464 Energy calculations suggest that only at high doses of strontium, comparable to those found for calcium, can strontium substitute for calcium. 16815919 8 7 theme large 1321:1325 arg1 area 1327:1330 a large area 1319:1330 a large area that includes loops 210-228 and 250-297 1319:1370 Since calcium helps stabilize a large area that includes loops 210-228 and 250-297, its substitution by strontium could affect the stability of this region. 16815919 1 8 theme painful 222:228 arg1 metastases 245:254 painful scattered bone metastases 222:254 painful scattered bone metastases 222:254 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 5 9 theme physiological 916:928 arg1 implications 930:941 important physiological implications 906:941 important physiological implications 906:941 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 1 10 theme scattered 230:238 arg1 metastases 245:254 painful scattered bone metastases 222:254 painful scattered bone metastases 222:254 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 0 11 dep implication 76:86 arg1 studies 11:17 Structural studies 0:17 Structural studies of human alkaline phosphatase in complex with strontium 0:73 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 7 12 theme polarizable 1092:1102 arg1 TCPEp 1116:1120 the flexible and polarizable force-field TCPEp 1075:1120 the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) 1075:1182 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 13 theme interaction 1231:1241 arg1 energies 1243:1250 similar interaction energies 1223:1250 similar interaction energies 1223:1250 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 1 14 theme bone 240:243 arg1 metastases 245:254 painful scattered bone metastases 222:254 painful scattered bone metastases 222:254 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 5 15 theme phosphatase 824:834 arg1 structure 764:772 the crystal structure 752:772 the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications 752:941 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 2 16 theme bone 373:376 arg1 mineralization 378:391 bone mineralization 373:391 bone mineralization 373:391 At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. 16815919 1 17 used used 136:139 arg2 Strontium 123:131 Strontium 123:131 Strontium 123:131 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 1 17 used used 136:139 arg2 isotope 259:265 isotope 259:265 isotope 259:265 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 6 18 theme coordination 1050:1061 arg1 modification 1024:1035 concomitant modification 1012:1035 concomitant modification of the metal coordination 1012:1061 The structure shows that strontium substitutes the calcium ion with concomitant modification of the metal coordination. 16815919 3 19 theme osteomalacia 398:409 arg1 symptoms 411:418 The osteomalacia symptoms 394:418 The osteomalacia symptoms 394:418 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 0 20 theme secondary 96:104 arg1 effect 106:111 its secondary effect 92:111 its secondary effect 92:111 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 7 21 theme flexible 1079:1086 arg1 TCPEp 1116:1120 the flexible and polarizable force-field TCPEp 1075:1120 the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) 1075:1182 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 5 22 theme strontium-substituted 777:797 arg1 isozyme 854:860 a related isozyme 844:860 a related isozyme of TNAP 844:868 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 22 theme strontium-substituted 777:797 arg1 PLAP 837:840 PLAP 837:840 PLAP 837:840 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 22 theme strontium-substituted 777:797 arg1 phosphatase 824:834 human placental alkaline phosphatase 799:834 strontium-substituted human placental alkaline phosphatase (PLAP) 777:841 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 10 23 theme high 1639:1642 arg1 doses 1644:1648 high doses 1639:1648 high doses of strontium 1639:1661 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 3 24 theme alkaline 552:559 arg1 TNAP 574:577 TNAP 574:577 TNAP 574:577 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 3 24 theme alkaline 552:559 arg1 phosphatase 561:571 tissue-nonspecific alkaline phosphatase 533:571 tissue-nonspecific alkaline phosphatase (TNAP) 533:578 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 1 25 theme osteoporosis 161:172 arg1 treatment 148:156 the treatment 144:156 the treatment of osteoporosis as a ranelate compound 144:195 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 0 26 theme Structural 0:9 arg1 studies 11:17 Structural studies 0:17 Structural studies of human alkaline phosphatase in complex with strontium 0:73 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 1 27 theme metastases 245:254 arg1 treatment 209:217 the treatment 205:217 the treatment of painful scattered bone metastases 205:254 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 2 28 theme high 276:279 arg1 doses 281:285 very high doses 271:285 very high doses 271:285 At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. 16815919 5 29 theme related 846:852 arg1 isozyme 854:860 a related isozyme 844:860 a related isozyme of TNAP 844:868 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 29 theme related 846:852 arg1 phosphatase 824:834 human placental alkaline phosphatase 799:834 strontium-substituted human placental alkaline phosphatase (PLAP) 777:841 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 0 30 theme human 22:26 arg1 phosphatase 37:47 human alkaline phosphatase 22:47 human alkaline phosphatase 22:47 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 10 31 theme alkaline 1664:1671 arg1 phosphatase 1673:1683 alkaline phosphatase 1664:1683 alkaline phosphatase 1664:1683 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 3 32 from mutations 498:506 arg1 gene 515:518 the gene 511:518 the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP) 511:578 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 5 33 theme important 906:914 arg1 implications 930:941 important physiological implications 906:941 important physiological implications 906:941 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 2 34 theme mineralization 378:391 arg1 impairment 359:368 impairment 359:368 impairment of bone mineralization 359:391 At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. 16815919 5 35 theme placental 805:813 arg1 isozyme 854:860 a related isozyme 844:860 a related isozyme of TNAP 844:868 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 35 theme placental 805:813 arg1 PLAP 837:840 PLAP 837:840 PLAP 837:840 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 35 theme placental 805:813 arg1 phosphatase 824:834 human placental alkaline phosphatase 799:834 strontium-substituted human placental alkaline phosphatase (PLAP) 777:841 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 10 36 theme targets 1712:1718 arg1 one 1701:1703 one 1701:1703 one 1701:1703 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 10 36 theme targets 1712:1718 arg1 targets 1712:1718 the targets 1708:1718 the targets of strontium 1708:1731 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 0 37 from implication 76:86 arg1 bones 116:120 bones 116:120 bones 116:120 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 3 38 theme tissue-nonspecific 533:550 arg1 TNAP 574:577 TNAP 574:577 TNAP 574:577 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 3 38 theme tissue-nonspecific 533:550 arg1 phosphatase 561:571 tissue-nonspecific alkaline phosphatase 533:571 tissue-nonspecific alkaline phosphatase (TNAP) 533:578 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 5 39 contain have 901:904 arg2 implications 930:941 important physiological implications 906:941 important physiological implications 906:941 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 39 contain have 901:904 arg1 replacement 885:895 such replacement 880:895 such replacement 880:895 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 1 40 theme ranelate 179:186 arg1 compound 188:195 a ranelate compound 177:195 a ranelate compound 177:195 Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. 16815919 0 41 theme phosphatase 37:47 arg1 studies 11:17 Structural studies 0:17 Structural studies of human alkaline phosphatase in complex with strontium 0:73 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 3 42 theme inherited 463:471 arg1 disorder 473:480 a rare inherited disorder 456:480 a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP) 456:578 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 3 42 theme inherited 463:471 arg1 hypophosphatasia 438:453 hypophosphatasia 438:453 hypophosphatasia 438:453 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 9 43 theme high 1487:1490 arg1 doses 1492:1496 high doses 1487:1496 high doses of strontium 1487:1509 Energy calculations suggest that only at high doses of strontium, comparable to those found for calcium, can strontium substitute for calcium. 16815919 6 44 theme metal 1044:1048 arg1 coordination 1050:1061 the metal coordination 1040:1061 the metal coordination 1040:1061 The structure shows that strontium substitutes the calcium ion with concomitant modification of the metal coordination. 16815919 10 45 theme strontium 1653:1661 arg1 doses 1644:1648 high doses 1639:1648 high doses of strontium 1639:1661 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 0 46 theme alkaline 28:35 arg1 phosphatase 37:47 human alkaline phosphatase 22:47 human alkaline phosphatase 22:47 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 7 47 theme classical 1139:1147 arg1 effects 1162:1168 classical polarization effects 1139:1168 classical polarization effects 1139:1168 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 5 48 theme human 799:803 arg1 isozyme 854:860 a related isozyme 844:860 a related isozyme of TNAP 844:868 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 48 theme human 799:803 arg1 PLAP 837:840 PLAP 837:840 PLAP 837:840 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 5 48 theme human 799:803 arg1 phosphatase 824:834 human placental alkaline phosphatase 799:834 strontium-substituted human placental alkaline phosphatase (PLAP) 777:841 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 4 49 theme binding 625:631 arg1 sites--two 633:642 four metal binding sites--two 614:642 four metal binding sites--two for zinc 614:651 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 5 50 theme crystal 756:762 arg1 structure 764:772 the crystal structure 752:772 the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications 752:941 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 7 51 dep TCPEp 1116:1120 arg1 effects 1162:1168 classical polarization effects 1139:1168 classical polarization effects 1139:1168 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 51 dep TCPEp 1116:1120 arg1 topological 1123:1133 topological 1123:1133 topological 1123:1133 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 3 52 theme rare 458:461 arg1 disorder 473:480 a rare inherited disorder 456:480 a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP) 456:578 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 3 52 theme rare 458:461 arg1 hypophosphatasia 438:453 hypophosphatasia 438:453 hypophosphatasia 438:453 The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). 16815919 6 53 with ion 1003:1005 arg1 modification 1024:1035 concomitant modification 1012:1035 concomitant modification of the metal coordination 1012:1061 The structure shows that strontium substitutes the calcium ion with concomitant modification of the metal coordination. 16815919 2 54 theme certain 294:300 arg1 conditions 302:311 certain conditions 294:311 certain conditions 294:311 At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. 16815919 7 55 theme polarization 1149:1160 arg1 effects 1162:1168 classical polarization effects 1139:1168 classical polarization effects 1139:1168 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 10 56 theme strontium 1723:1731 arg1 targets 1712:1718 the targets 1708:1718 the targets of strontium 1708:1731 Since osteomalacia is observed after ingestion of high doses of strontium, alkaline phosphatase is likely to be one of the targets of strontium, and thus this enzyme might be involved in this disease. 16815919 5 57 theme such 880:883 arg1 replacement 885:895 such replacement 880:895 such replacement 880:895 Here we present the crystal structure of strontium-substituted human placental alkaline phosphatase (PLAP), a related isozyme of TNAP, in which such replacement can have important physiological implications. 16815919 4 58 theme alkaline 587:594 arg1 phosphatases 596:607 Human alkaline phosphatases 581:607 Human alkaline phosphatases 581:607 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 7 59 contain has 1219:1221 arg1 strontium 1209:1217 strontium 1209:1217 strontium 1209:1217 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 59 contain has 1219:1221 arg1 calcium 1198:1204 calcium 1198:1204 calcium 1198:1204 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 59 contain has 1219:1221 arg2 energies 1243:1250 similar interaction energies 1223:1250 similar interaction energies 1223:1250 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 8 60 theme region 1438:1443 arg1 stability 1420:1428 the stability 1416:1428 the stability of this region 1416:1443 Since calcium helps stabilize a large area that includes loops 210-228 and 250-297, its substitution by strontium could affect the stability of this region. 16815919 4 61 contain have 609:612 arg2 sites--two 633:642 four metal binding sites--two 614:642 four metal binding sites--two for zinc 614:651 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 4 61 contain have 609:612 arg1 phosphatases 596:607 Human alkaline phosphatases 581:607 Human alkaline phosphatases 581:607 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 0 62 with complex 52:58 arg1 strontium 65:73 strontium 65:73 strontium 65:73 Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones. 16815919 4 63 theme calcium 685:691 arg1 ion--that 693:701 calcium ion--that 685:701 calcium ion--that 685:701 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 4 64 theme Human 581:585 arg1 phosphatases 596:607 Human alkaline phosphatases 581:607 Human alkaline phosphatases 581:607 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 6 65 theme concomitant 1012:1022 arg1 modification 1024:1035 concomitant modification 1012:1035 concomitant modification of the metal coordination 1012:1061 The structure shows that strontium substitutes the calcium ion with concomitant modification of the metal coordination. 16815919 4 66 theme metal 619:623 arg1 sites--two 633:642 four metal binding sites--two 614:642 four metal binding sites--two for zinc 614:651 Human alkaline phosphatases have four metal binding sites--two for zinc, one for magnesium, and one for calcium ion--that can be substituted by strontium. 16815919 9 67 theme strontium 1501:1509 arg1 doses 1492:1496 high doses 1487:1496 high doses of strontium 1487:1509 Energy calculations suggest that only at high doses of strontium, comparable to those found for calcium, can strontium substitute for calcium. 16815919 6 68 theme calcium 995:1001 arg1 ion 1003:1005 the calcium ion 991:1005 the calcium ion with concomitant modification of the metal coordination 991:1061 The structure shows that strontium substitutes the calcium ion with concomitant modification of the metal coordination. 16815919 7 69 theme similar 1223:1229 arg1 energies 1243:1250 similar interaction energies 1223:1250 similar interaction energies 1223:1250 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 70 theme PLAP 1283:1286 arg1 site 1275:1278 the calcium-binding site 1255:1278 the calcium-binding site of PLAP 1255:1286 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 70 theme PLAP 1283:1286 arg1 PLAP 1283:1286 PLAP 1283:1286 PLAP 1283:1286 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 71 theme calcium-binding 1259:1273 arg1 site 1275:1278 the calcium-binding site 1255:1278 the calcium-binding site of PLAP 1255:1286 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 16815919 7 71 theme calcium-binding 1259:1273 arg1 PLAP 1283:1286 PLAP 1283:1286 PLAP 1283:1286 The use of the flexible and polarizable force-field TCPEp (topological and classical polarization effects for proteins) predicts that calcium or strontium has similar interaction energies at the calcium-binding site of PLAP. 15946677 7 0 theme human 1136:1140 arg1 phosphatase 1151:1161 human alkaline phosphatase 1136:1161 the human alkaline phosphatase family 1132:1168 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 4 1 from first 740:744 arg1 complex 776:782 complex 776:782 complex with organic inhibitors 776:806 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 6 2 theme located 1036:1042 arg1 site 1031:1034 a remote peripheral site 1011:1034 a remote peripheral site located 28A from the active site 1011:1067 In contrast, all three ligands interact at a remote peripheral site located 28A from the active site. 15946677 0 3 theme functional 75:84 arg1 ligands 86:92 functional ligands 75:92 functional ligands 75:92 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 0 4 from studies 11:17 arg1 complex 62:68 complex 62:68 complex with functional ligands 62:92 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 8 5 theme peripheral 1410:1419 arg1 site 1429:1432 the peripheral binding site 1406:1432 the peripheral binding site 1406:1432 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 6 6 theme active 1057:1062 arg1 site 1064:1067 the active site 1053:1067 the active site 1053:1067 In contrast, all three ligands interact at a remote peripheral site located 28A from the active site. 15946677 2 7 theme reduced 498:504 arg1 rate 538:541 a reduced and better-regulated hydrolytic rate 496:541 a reduced and better-regulated hydrolytic rate 496:541 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 1 8 from effects 279:285 arg1 proliferation 309:321 fibroblast proliferation 298:321 fibroblast proliferation 298:321 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 2 9 attach linked 448:453 arg1 function 473:480 its biological function 458:480 its biological function that requires a reduced and better-regulated hydrolytic rate 458:541 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 2 9 attach linked 448:453 arg2 regulation 430:439 such regulation 425:439 such regulation 425:439 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 8 10 theme crucial 1289:1295 arg1 role 1297:1300 the crucial role 1285:1300 the crucial role played by position 429 1285:1323 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 3 11 theme 1.9A 676:679 arg1 resolution 681:690 1.9A resolution 676:690 1.9A resolution 676:690 To understand how such disparate ligands are able to inhibit the enzyme, we solved the structure of the complexes at 1.6A, 1.9A and 1.9A resolution, respectively. 15946677 5 12 theme structural 911:920 arg1 data 922:925 structural data 911:925 structural data on the uncompetitive inhibition process 911:965 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 4 13 theme crystal 713:719 arg1 structures 721:730 These crystal structures 707:730 These crystal structures 707:730 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 4 13 theme crystal 713:719 arg1 first 740:744 first 740:744 first 740:744 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 5 14 theme active 869:874 arg1 pocket 893:898 the active site hydrophobic pocket 865:898 the active site hydrophobic pocket 865:898 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 7 15 theme other 1115:1119 arg1 members 1121:1127 the other members 1111:1127 the other members of the human alkaline phosphatase family 1111:1168 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 8 16 theme binding 1421:1427 arg1 site 1429:1432 the peripheral binding site 1406:1432 the peripheral binding site 1406:1432 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 0 17 with complex 62:68 arg1 ligands 86:92 functional ligands 75:92 functional ligands 75:92 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 6 18 theme peripheral 1020:1029 arg1 site 1031:1034 a remote peripheral site 1011:1034 a remote peripheral site located 28A from the active site 1011:1067 In contrast, all three ligands interact at a remote peripheral site located 28A from the active site. 15946677 0 19 theme phosphatase 47:57 arg1 studies 11:17 Structural studies 0:17 Structural studies of human placental alkaline phosphatase in complex with functional ligands 0:92 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 4 20 theme organic 789:795 arg1 inhibitors 797:806 organic inhibitors 789:806 organic inhibitors 789:806 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 8 21 theme isozyme 1495:1501 arg1 specialization 1468:1481 the functional specialization 1453:1481 the functional specialization of the PLAP isozyme 1453:1501 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 6 22 theme remote 1013:1018 arg1 site 1031:1034 a remote peripheral site 1011:1034 a remote peripheral site located 28A from the active site 1011:1067 In contrast, all three ligands interact at a remote peripheral site located 28A from the active site. 15946677 2 23 theme such 425:428 arg1 regulation 430:439 such regulation 425:439 such regulation 425:439 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 7 24 theme alkaline 1142:1149 arg1 phosphatase 1151:1161 human alkaline phosphatase 1136:1161 the human alkaline phosphatase family 1132:1168 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 8 25 theme PLAP 1490:1493 arg1 isozyme 1495:1501 the PLAP isozyme 1486:1501 the PLAP isozyme 1486:1501 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 5 26 theme site 876:879 arg1 pocket 893:898 the active site hydrophobic pocket 865:898 the active site hydrophobic pocket 865:898 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 0 27 theme Structural 0:9 arg1 studies 11:17 Structural studies 0:17 Structural studies of human placental alkaline phosphatase in complex with functional ligands 0:92 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 1 28 from antagonist 261:270 arg1 proliferation 309:321 fibroblast proliferation 298:321 fibroblast proliferation 298:321 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 5 29 theme hydrophobic 881:891 arg1 pocket 893:898 the active site hydrophobic pocket 865:898 the active site hydrophobic pocket 865:898 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 0 30 theme human 22:26 arg1 phosphatase 47:57 human placental alkaline phosphatase 22:57 human placental alkaline phosphatase 22:57 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 5 31 theme inhibition 948:957 arg1 process 959:965 the uncompetitive inhibition process 930:965 the uncompetitive inhibition process 930:965 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 8 32 from site 1339:1342 arg1 modulation 1351:1360 the modulation 1347:1360 the modulation of the catalytic process 1347:1385 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 7 33 theme isozymes 1222:1229 arg1 structures 1192:1201 the structures 1188:1201 the structures of the other human isozymes 1188:1229 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 8 34 theme functional 1457:1466 arg1 specialization 1468:1481 the functional specialization 1453:1481 the functional specialization of the PLAP isozyme 1453:1501 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 2 35 theme hydrolytic 527:536 arg1 rate 538:541 a reduced and better-regulated hydrolytic rate 496:541 a reduced and better-regulated hydrolytic rate 496:541 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 1 36 theme effectors 187:195 arg1 l-phenylalanine 205:219 l-phenylalanine 205:219 l-phenylalanine 205:219 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 36 theme effectors 187:195 arg1 number 177:182 a number 175:182 a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue 175:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 0 37 theme alkaline 38:45 arg1 phosphatase 47:57 human placental alkaline phosphatase 22:57 human placental alkaline phosphatase 22:57 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 2 38 theme better-regulated 510:525 arg1 rate 538:541 a reduced and better-regulated hydrolytic rate 496:541 a reduced and better-regulated hydrolytic rate 496:541 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 6 39 dep located 1036:1042 arg1 28A 1044:1046 28A 1044:1046 28A 1044:1046 In contrast, all three ligands interact at a remote peripheral site located 28A from the active site. 15946677 7 40 theme other 1210:1214 arg1 isozymes 1222:1229 the other human isozymes 1206:1229 the other human isozymes 1206:1229 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 0 41 theme placental 28:36 arg1 phosphatase 47:57 human placental alkaline phosphatase 22:57 human placental alkaline phosphatase 22:57 Structural studies of human placental alkaline phosphatase in complex with functional ligands. 15946677 1 42 theme effects 279:285 arg1 antagonist 261:270 an antagonist 258:270 an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue 258:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 42 theme effects 279:285 arg1 inhibitor 239:247 an uncompetitive inhibitor 222:247 an uncompetitive inhibitor 222:247 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 43 theme non-hydrolysable 369:384 arg1 p-nitrophenyl-phosphonate 330:354 p-nitrophenyl-phosphonate 330:354 p-nitrophenyl-phosphonate (PNPPate) 330:364 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 43 theme non-hydrolysable 369:384 arg1 analogue 396:403 a non-hydrolysable substrate analogue 367:403 a non-hydrolysable substrate analogue 367:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 4 44 theme phosphatase 761:771 arg1 structures 721:730 These crystal structures 707:730 These crystal structures 707:730 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 4 44 theme phosphatase 761:771 arg1 first 740:744 first 740:744 first 740:744 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 7 45 theme human 1216:1220 arg1 isozymes 1222:1229 the other human isozymes 1206:1229 the other human isozymes 1206:1229 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 1 46 theme substrate 386:394 arg1 p-nitrophenyl-phosphonate 330:354 p-nitrophenyl-phosphonate 330:354 p-nitrophenyl-phosphonate (PNPPate) 330:364 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 46 theme substrate 386:394 arg1 analogue 396:403 a non-hydrolysable substrate analogue 367:403 a non-hydrolysable substrate analogue 367:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 3 47 theme such 562:565 arg1 ligands 577:583 such disparate ligands 562:583 such disparate ligands 562:583 To understand how such disparate ligands are able to inhibit the enzyme, we solved the structure of the complexes at 1.6A, 1.9A and 1.9A resolution, respectively. 15946677 4 48 from complex 776:782 arg1 structures 721:730 These crystal structures 707:730 These crystal structures 707:730 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 4 48 from complex 776:782 arg1 first 740:744 first 740:744 first 740:744 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 1 49 theme human 111:115 arg1 PLAP 149:152 PLAP 149:152 PLAP 149:152 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 49 theme human 111:115 arg1 phosphatase 136:146 human placental alkaline phosphatase (PLAP) 111:153 human placental alkaline phosphatase (PLAP) 111:153 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 50 from proliferation 309:321 arg1 antagonist 261:270 an antagonist 258:270 an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue 258:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 50 from proliferation 309:321 arg1 inhibitor 239:247 an uncompetitive inhibitor 222:247 an uncompetitive inhibitor 222:247 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 51 theme PLAP 290:293 arg1 effects 279:285 the effects 275:285 the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue 275:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 8 52 theme process 1379:1385 arg1 modulation 1351:1360 the modulation 1347:1360 the modulation of the catalytic process 1347:1385 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 3 53 theme disparate 567:575 arg1 ligands 577:583 such disparate ligands 562:583 such disparate ligands 562:583 To understand how such disparate ligands are able to inhibit the enzyme, we solved the structure of the complexes at 1.6A, 1.9A and 1.9A resolution, respectively. 15946677 1 54 theme placental 117:125 arg1 PLAP 149:152 PLAP 149:152 PLAP 149:152 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 54 theme placental 117:125 arg1 phosphatase 136:146 human placental alkaline phosphatase (PLAP) 111:153 human placental alkaline phosphatase (PLAP) 111:153 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 3 55 theme complexes 648:656 arg1 structure 631:639 the structure 627:639 the structure of the complexes 627:656 To understand how such disparate ligands are able to inhibit the enzyme, we solved the structure of the complexes at 1.6A, 1.9A and 1.9A resolution, respectively. 15946677 8 56 theme catalytic 1369:1377 arg1 process 1379:1385 the catalytic process 1365:1385 the catalytic process 1365:1385 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 8 57 theme active 1332:1337 arg1 site 1339:1342 the active site 1328:1342 the active site in the modulation of the catalytic process 1328:1385 This comparison highlights the crucial role played by position 429 at the active site in the modulation of the catalytic process, and suggests that the peripheral binding site may be involved in the functional specialization of the PLAP isozyme. 15946677 6 58 from site 1064:1067 arg1 located 1036:1042 located 1036:1042 located 1036:1042 In contrast, all three ligands interact at a remote peripheral site located 28A from the active site. 15946677 1 59 theme alkaline 127:134 arg1 PLAP 149:152 PLAP 149:152 PLAP 149:152 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 59 theme alkaline 127:134 arg1 phosphatase 136:146 human placental alkaline phosphatase (PLAP) 111:153 human placental alkaline phosphatase (PLAP) 111:153 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 5 60 theme uncompetitive 934:946 arg1 process 959:965 the uncompetitive inhibition process 930:965 the uncompetitive inhibition process 930:965 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 1 61 theme phosphatase 136:146 arg1 activity 99:106 The activity 95:106 The activity of human placental alkaline phosphatase (PLAP) 95:153 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 4 62 with complex 776:782 arg1 inhibitors 797:806 organic inhibitors 789:806 organic inhibitors 789:806 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 7 63 theme phosphatase 1151:1161 arg1 family 1163:1168 the human alkaline phosphatase family 1132:1168 the human alkaline phosphatase family 1132:1168 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 4 64 theme alkaline 752:759 arg1 phosphatase 761:771 an alkaline phosphatase 749:771 an alkaline phosphatase 749:771 These crystal structures are the first of an alkaline phosphatase in complex with organic inhibitors. 15946677 7 65 theme family 1163:1168 arg1 members 1121:1127 the other members 1111:1127 the other members of the human alkaline phosphatase family 1111:1168 In order to extend these observations to the other members of the human alkaline phosphatase family, we have modelled the structures of the other human isozymes and compared them to PLAP. 15946677 2 66 theme biological 462:471 arg1 function 473:480 its biological function 458:480 its biological function that requires a reduced and better-regulated hydrolytic rate 458:541 For the first two, such regulation may be linked to its biological function that requires a reduced and better-regulated hydrolytic rate. 15946677 1 67 theme fibroblast 298:307 arg1 proliferation 309:321 fibroblast proliferation 298:321 fibroblast proliferation 298:321 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 5 68 from data 922:925 arg1 process 959:965 the uncompetitive inhibition process 930:965 the uncompetitive inhibition process 930:965 Of the three inhibitors, only l-Phe and PNPPate bind at the active site hydrophobic pocket, providing structural data on the uncompetitive inhibition process. 15946677 1 69 theme uncompetitive 225:237 arg1 l-phenylalanine 205:219 l-phenylalanine 205:219 l-phenylalanine 205:219 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 69 theme uncompetitive 225:237 arg1 5'-AMP 250:255 5'-AMP 250:255 5'-AMP 250:255 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 69 theme uncompetitive 225:237 arg1 antagonist 261:270 an antagonist 258:270 an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue 258:403 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 15946677 1 69 theme uncompetitive 225:237 arg1 inhibitor 239:247 an uncompetitive inhibitor 222:247 an uncompetitive inhibitor 222:247 The activity of human placental alkaline phosphatase (PLAP) is downregulated by a number of effectors such as l-phenylalanine, an uncompetitive inhibitor, 5'-AMP, an antagonist of the effects of PLAP on fibroblast proliferation and by p-nitrophenyl-phosphonate (PNPPate), a non-hydrolysable substrate analogue. 20693656 0 0 theme interactions 82:93 arg1 pattern 71:77 a consistent pattern 58:77 a consistent pattern of interactions at the peripheral site 58:116 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 5 1 theme omitted 586:592 arg1 chains 599:604 omitted side chains 586:604 omitted side chains 586:604 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 5 2 theme multiple 476:483 arg1 corrections 485:495 The multiple corrections 472:495 The multiple corrections to the model 472:508 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 7 3 contain contained 787:795 arg2 p-nitrophenol 802:814 only p-nitrophenol 797:814 only p-nitrophenol in three distinct sites 797:838 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 3 contain contained 787:795 arg1 structure 767:775 The structure 763:775 The structure with pNPP 763:785 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 8 4 from site 1043:1046 arg1 interactions 1012:1023 interactions 1012:1023 interactions at the peripheral site 1012:1046 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 8 4 from site 1043:1046 arg1 pattern 1001:1007 a consistent pattern 988:1007 a consistent pattern of interactions at the peripheral site 988:1046 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 4 5 theme previous 455:462 arg1 report 464:469 the previous report 451:469 the previous report 451:469 Significant variations in ligand positioning and identity were found compared with the previous report. 20693656 5 6 theme side 594:597 arg1 chains 599:604 omitted side chains 586:604 omitted side chains 586:604 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 7 7 theme p-nitrophenyl 887:899 arg1 group 901:905 the p-nitrophenyl group 883:905 the p-nitrophenyl group in two of the sites instead of 5'-AMP 883:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 8 from group 901:905 arg1 two 910:912 two 910:912 two 910:912 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 8 from group 901:905 arg1 sites 921:925 the sites 917:925 the sites instead of 5'-AMP 917:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 8 from group 901:905 arg1 5'-AMP 938:943 5'-AMP 938:943 5'-AMP 938:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 9 theme only 797:800 arg1 p-nitrophenol 802:814 only p-nitrophenol 797:814 only p-nitrophenol in three distinct sites 797:838 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 1 10 theme enzyme 241:246 arg1 structures 223:232 the structures 219:232 the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 219:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 10 theme enzyme 241:246 arg1 phosphatase 206:216 human placental alkaline phosphatase 181:216 human placental alkaline phosphatase 181:216 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 7 11 contain contained 873:881 arg2 group 901:905 the p-nitrophenyl group 883:905 the p-nitrophenyl group in two of the sites instead of 5'-AMP 883:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 11 contain contained 873:881 arg1 structure 851:859 the structure 847:859 the structure with 5'-AMP 847:871 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 1 12 dep regulators 266:275 arg1 L-Phe 277:281 L-Phe 277:281 L-Phe 277:281 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 12 dep regulators 266:275 arg1 regulators 266:275 the putative regulators 253:275 the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 253:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 12 dep regulators 266:275 arg1 2005 316:319 2005 316:319 2005 316:319 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 12 dep regulators 266:275 arg1 pNPP 284:287 pNPP 284:287 pNPP 284:287 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 12 dep regulators 266:275 arg1 [Llinas 300:306 5'-AMP [Llinas 293:306 5'-AMP [Llinas et al. (2005) 293:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 5 13 theme chains 599:604 arg1 modeling 574:581 the modeling 570:581 the modeling of omitted side chains and multiple disordered residues 570:637 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 5 14 theme electron-density 538:553 arg1 maps 555:558 the electron-density maps 534:558 the electron-density maps 534:558 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 7 15 theme 5'-AMP 938:943 arg1 sites 921:925 the sites 917:925 the sites instead of 5'-AMP 917:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 15 theme 5'-AMP 938:943 arg1 5'-AMP 938:943 5'-AMP 938:943 5'-AMP 938:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 16 from p-nitrophenol 802:814 arg1 sites 834:838 three distinct sites 819:838 three distinct sites 819:838 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 1 17 dep [Llinas 300:306 arg1 al 311:312 et al 308:312 5'-AMP [Llinas et al. (2005) 293:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 3 18 dep 350 337:339 arg1 441-451 342:348 441-451 342:348 441-451 342:348 350, 441-451] were re-refined. 20693656 0 19 theme peripheral 102:111 arg1 site 113:116 the peripheral site 98:116 the peripheral site 98:116 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 8 20 theme re-refined 964:973 arg1 models 975:980 the re-refined models 960:980 the re-refined models 960:980 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 1 21 theme functional 161:170 arg1 sites 172:176 the functional sites 157:176 the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol 157:328 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 21 theme functional 161:170 arg1 phosphatase 206:216 human placental alkaline phosphatase 181:216 human placental alkaline phosphatase 181:216 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 6 22 theme binding 718:724 arg1 site 726:729 the peripheral binding site 703:729 the peripheral binding site 703:729 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 0 23 theme Refined 0:6 arg1 structures 8:17 Refined structures 0:17 Refined structures of placental alkaline phosphatase 0:51 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 1 24 theme putative 257:264 arg1 L-Phe 277:281 L-Phe 277:281 L-Phe 277:281 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 24 theme putative 257:264 arg1 regulators 266:275 the putative regulators 253:275 the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 253:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 24 theme putative 257:264 arg1 pNPP 284:287 pNPP 284:287 pNPP 284:287 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 24 theme putative 257:264 arg1 [Llinas 300:306 5'-AMP [Llinas 293:306 5'-AMP [Llinas et al. (2005) 293:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 6 25 theme peripheral 707:716 arg1 site 726:729 the peripheral binding site 703:729 the peripheral binding site 703:729 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 7 26 with structure 767:775 arg1 pNPP 782:785 pNPP 782:785 pNPP 782:785 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 0 27 theme placental 22:30 arg1 phosphatase 41:51 placental alkaline phosphatase 22:51 placental alkaline phosphatase 22:51 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 7 28 with structure 851:859 arg1 5'-AMP 866:871 5'-AMP 866:871 5'-AMP 866:871 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 8 29 theme models 975:980 arg1 Comparison 946:955 Comparison 946:955 Comparison of the re-refined models 946:980 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 1 30 theme human 181:185 arg1 structures 223:232 the structures 219:232 the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 219:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 30 theme human 181:185 arg1 Mol 326:328 Mol 326:328 Mol 326:328 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 30 theme human 181:185 arg1 phosphatase 206:216 human placental alkaline phosphatase 181:216 human placental alkaline phosphatase 181:216 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 8 31 theme consistent 990:999 arg1 pattern 1001:1007 a consistent pattern 988:1007 a consistent pattern of interactions at the peripheral site 988:1046 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 1 32 theme placental 187:195 arg1 structures 223:232 the structures 219:232 the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 219:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 32 theme placental 187:195 arg1 Mol 326:328 Mol 326:328 Mol 326:328 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 32 theme placental 187:195 arg1 phosphatase 206:216 human placental alkaline phosphatase 181:216 human placental alkaline phosphatase 181:216 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 0 33 theme phosphatase 41:51 arg1 structures 8:17 Refined structures 0:17 Refined structures of placental alkaline phosphatase 0:51 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 8 34 theme interactions 1012:1023 arg1 pattern 1001:1007 a consistent pattern 988:1007 a consistent pattern of interactions at the peripheral site 988:1046 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 6 35 from site 726:729 arg1 change 668:673 a change 666:673 a change 666:673 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 6 35 from site 726:729 arg1 position 682:689 the position 678:689 the position of L-Phe at the peripheral binding site 678:729 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 1 36 theme alkaline 197:204 arg1 structures 223:232 the structures 219:232 the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 219:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 36 theme alkaline 197:204 arg1 Mol 326:328 Mol 326:328 Mol 326:328 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 36 theme alkaline 197:204 arg1 phosphatase 206:216 human placental alkaline phosphatase 181:216 human placental alkaline phosphatase 181:216 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 0 37 theme alkaline 32:39 arg1 phosphatase 41:51 placental alkaline phosphatase 22:51 placental alkaline phosphatase 22:51 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 4 38 from variations 380:389 arg1 identity 417:424 identity 417:424 identity 417:424 Significant variations in ligand positioning and identity were found compared with the previous report. 20693656 4 38 from variations 380:389 arg1 positioning 401:411 ligand positioning 394:411 ligand positioning 394:411 Significant variations in ligand positioning and identity were found compared with the previous report. 20693656 0 39 from pattern 71:77 arg1 site 113:116 the peripheral site 98:116 the peripheral site 98:116 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 1 40 theme phosphatase 206:216 arg1 sites 172:176 the functional sites 157:176 the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol 157:328 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 40 theme phosphatase 206:216 arg1 phosphatase 206:216 human placental alkaline phosphatase 181:216 human placental alkaline phosphatase 181:216 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 5 41 theme multiple 610:617 arg1 residues 630:637 multiple disordered residues 610:637 multiple disordered residues 610:637 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 4 42 theme ligand 394:399 arg1 positioning 401:411 ligand positioning 394:411 ligand positioning 394:411 Significant variations in ligand positioning and identity were found compared with the previous report. 20693656 6 43 theme L-Phe 694:698 arg1 position 682:689 the position 678:689 the position of L-Phe at the peripheral binding site 678:729 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 1 44 theme 5'-AMP 293:298 arg1 regulators 266:275 the putative regulators 253:275 the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 253:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 44 theme 5'-AMP 293:298 arg1 2005 316:319 2005 316:319 2005 316:319 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 44 theme 5'-AMP 293:298 arg1 [Llinas 300:306 5'-AMP [Llinas 293:306 5'-AMP [Llinas et al. (2005) 293:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 5 45 theme disordered 619:628 arg1 residues 630:637 multiple disordered residues 610:637 multiple disordered residues 610:637 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 0 46 from site 113:116 arg1 interactions 82:93 interactions 82:93 interactions at the peripheral site 82:116 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 0 46 from site 113:116 arg1 pattern 71:77 a consistent pattern 58:77 a consistent pattern of interactions at the peripheral site 58:116 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 7 47 theme sites 921:925 arg1 two 910:912 two 910:912 two 910:912 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 47 theme sites 921:925 arg1 sites 921:925 the sites 917:925 the sites instead of 5'-AMP 917:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 7 47 theme sites 921:925 arg1 5'-AMP 938:943 5'-AMP 938:943 5'-AMP 938:943 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 5 48 theme residues 630:637 arg1 modeling 574:581 the modeling 570:581 the modeling of omitted side chains and multiple disordered residues 570:637 The multiple corrections to the model improved the phases and the electron-density maps, allowing the modeling of omitted side chains and multiple disordered residues. 20693656 8 49 theme peripheral 1032:1041 arg1 site 1043:1046 the peripheral site 1028:1046 the peripheral site 1028:1046 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 4 50 theme Significant 368:378 arg1 variations 380:389 Significant variations 368:389 Significant variations in ligand positioning and identity 368:424 Significant variations in ligand positioning and identity were found compared with the previous report. 20693656 1 51 with structures 223:232 arg1 L-Phe 277:281 L-Phe 277:281 L-Phe 277:281 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 51 with structures 223:232 arg1 regulators 266:275 the putative regulators 253:275 the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005) 253:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 51 with structures 223:232 arg1 pNPP 284:287 pNPP 284:287 pNPP 284:287 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 1 51 with structures 223:232 arg1 [Llinas 300:306 5'-AMP [Llinas 293:306 5'-AMP [Llinas et al. (2005) 293:320 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 20693656 7 52 theme distinct 825:832 arg1 sites 834:838 three distinct sites 819:838 three distinct sites 819:838 The structure with pNPP contained only p-nitrophenol in three distinct sites, while the structure with 5'-AMP contained the p-nitrophenyl group in two of the sites instead of 5'-AMP. 20693656 6 53 from change 668:673 arg1 site 726:729 the peripheral binding site 703:729 the peripheral binding site 703:729 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 6 53 from change 668:673 arg1 position 682:689 the position 678:689 the position of L-Phe at the peripheral binding site 678:729 These improvements led to a change in the position of L-Phe at the peripheral binding site, which appeared to be reversed. 20693656 8 54 from pattern 1001:1007 arg1 site 1043:1046 the peripheral site 1028:1046 the peripheral site 1028:1046 Comparison of the re-refined models shows a consistent pattern of interactions at the peripheral site. 20693656 0 55 theme consistent 60:69 arg1 pattern 71:77 a consistent pattern 58:77 a consistent pattern of interactions at the peripheral site 58:116 Refined structures of placental alkaline phosphatase show a consistent pattern of interactions at the peripheral site. 20693656 1 56 theme deeper 136:141 arg1 insights 143:150 deeper insights 136:150 deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol 136:328 In order to gain deeper insights into the functional sites of human placental alkaline phosphatase, the structures of the enzyme with the putative regulators L-Phe, pNPP and 5'-AMP [Llinas et al. (2005), J. Mol. 17581592 5 0 with interaction 997:1007 arg1 antigen 1024:1030 the lipid antigen 1014:1030 the lipid antigen 1014:1030 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 7 1 theme lipid-antigen-presenting 1397:1420 arg1 molecule 1422:1429 a lipid-antigen-presenting molecule 1395:1429 a lipid-antigen-presenting molecule of the immune system 1395:1450 These findings provide direct insight into how a T-cell receptor recognizes a lipid-antigen-presenting molecule of the immune system. 17581592 2 2 theme killer 342:347 arg1 cells 357:361 natural killer T (NKT) cells 334:361 natural killer T (NKT) cells 334:361 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 2 theme killer 342:347 arg1 group 279:283 The most studied group 262:283 The most studied group of T cells that interact with lipid antigens 262:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 2 theme killer 342:347 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 4 3 theme potent 726:731 arg1 glycolipid 807:816 the archetypal CD1d-restricted glycolipid 776:816 the archetypal CD1d-restricted glycolipid 776:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 4 3 theme potent 726:731 arg1 alpha-galactosylceramide 750:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 1 4 theme T 253:253 arg1 cells 255:259 T cells 253:259 T cells 253:259 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 6 5 theme complexes 1308:1316 arg1 recognition 1274:1284 recognition 1274:1284 recognition of other CD1d-antigen complexes 1274:1316 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 6 6 theme beta-chain 1249:1258 arg1 variability 1222:1232 variability 1222:1232 variability of the NKT TCR beta-chain 1222:1258 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 1 7 theme MHC 162:164 arg1 molecules 180:188 non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules 111:188 molecules 180:188 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 4 8 from structure 663:671 arg1 complex 695:701 complex 695:701 complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid 695:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 6 9 theme innate 1182:1187 arg1 immunity 1189:1196 innate immunity 1182:1196 innate immunity 1182:1196 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 2 10 theme natural 334:340 arg1 cells 357:361 natural killer T (NKT) cells 334:361 natural killer T (NKT) cells 334:361 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 10 theme natural 334:340 arg1 group 279:283 The most studied group 262:283 The most studied group of T cells that interact with lipid antigens 262:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 10 theme natural 334:340 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 6 11 theme CD1d-antigen 1141:1152 arg1 complex 1154:1160 the CD1d-antigen complex 1137:1160 the CD1d-antigen complex that is typified in innate immunity 1137:1196 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 3 12 theme CD1d-antigen 535:546 arg1 complex 548:554 the CD1d-antigen complex 531:554 the CD1d-antigen complex 531:554 NKT-cell-mediated recognition of the CD1d-antigen complex has been implicated in microbial immunity, tumour immunity, autoimmunity and allergy. 17581592 6 13 theme NKT 1241:1243 arg1 beta-chain 1249:1258 the NKT TCR beta-chain 1237:1258 the NKT TCR beta-chain 1237:1258 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 5 14 theme CD1d-binding 943:954 arg1 cleft 956:960 the CD1d-binding cleft 939:960 the CD1d-binding cleft 939:960 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 4 15 theme TCR 688:690 arg1 structure 663:671 the structure 659:671 the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid 659:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 6 16 theme NKT 1113:1115 arg1 alpha-chain 1121:1131 the highly conserved NKT TCR alpha-chain 1092:1131 the highly conserved NKT TCR alpha-chain 1092:1131 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 1 17 theme CD1 78:80 arg1 family 82:87 The CD1 family 74:87 The CD1 family 74:87 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 1 17 theme CD1 78:80 arg1 cluster 100:106 a large cluster 92:106 a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells 92:259 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 5 18 theme receptor-peptide-antigen-MHC 841:868 arg1 complexes 870:878 T-cell receptor-peptide-antigen-MHC complexes 834:878 T-cell receptor-peptide-antigen-MHC complexes 834:878 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 2 19 theme NKT 431:433 arg1 receptor 421:428 a semi-invariant T-cell receptor 397:428 a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d 397:495 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 19 theme NKT 431:433 arg1 TCR 435:437 NKT TCR 431:437 NKT TCR 431:437 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 7 20 theme immune 1438:1443 arg1 system 1445:1450 the immune system 1434:1450 the immune system 1434:1450 These findings provide direct insight into how a T-cell receptor recognizes a lipid-antigen-presenting molecule of the immune system. 17581592 3 21 theme NKT-cell-mediated 498:514 arg1 recognition 516:526 NKT-cell-mediated recognition 498:526 NKT-cell-mediated recognition of the CD1d-antigen complex 498:554 NKT-cell-mediated recognition of the CD1d-antigen complex has been implicated in microbial immunity, tumour immunity, autoimmunity and allergy. 17581592 1 22 theme class-I-like 167:178 arg1 molecules 180:188 non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules 111:188 molecules 180:188 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 7 23 theme system 1445:1450 arg1 molecule 1422:1429 a lipid-antigen-presenting molecule 1395:1429 a lipid-antigen-presenting molecule of the immune system 1395:1450 These findings provide direct insight into how a T-cell receptor recognizes a lipid-antigen-presenting molecule of the immune system. 17581592 3 24 theme complex 548:554 arg1 recognition 516:526 NKT-cell-mediated recognition 498:526 NKT-cell-mediated recognition of the CD1d-antigen complex 498:554 NKT-cell-mediated recognition of the CD1d-antigen complex has been implicated in microbial immunity, tumour immunity, autoimmunity and allergy. 17581592 1 25 theme molecules 180:188 arg1 family 82:87 The CD1 family 74:87 The CD1 family 74:87 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 1 25 theme molecules 180:188 arg1 cluster 100:106 a large cluster 92:106 a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells 92:259 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 0 26 theme CD1d-lipid-antigen 0:17 arg1 recognition 19:29 CD1d-lipid-antigen recognition 0:29 CD1d-lipid-antigen recognition by the semi-invariant NKT T-cell receptor 0:71 CD1d-lipid-antigen recognition by the semi-invariant NKT T-cell receptor. 17581592 5 27 theme extreme 924:930 arg1 end 932:934 the extreme end 920:934 the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen 920:1030 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 7 28 theme direct 1342:1347 arg1 insight 1349:1355 direct insight 1342:1355 direct insight into how a T-cell receptor recognizes a lipid-antigen-presenting molecule of the immune system 1342:1450 These findings provide direct insight into how a T-cell receptor recognizes a lipid-antigen-presenting molecule of the immune system. 17581592 2 29 theme NKT 352:354 arg1 cells 357:361 natural killer T (NKT) cells 334:361 natural killer T (NKT) cells 334:361 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 29 theme NKT 352:354 arg1 group 279:283 The most studied group 262:283 The most studied group of T cells that interact with lipid antigens 262:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 29 theme NKT 352:354 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 30 theme T-cell 414:419 arg1 receptor 421:428 a semi-invariant T-cell receptor 397:428 a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d 397:495 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 30 theme T-cell 414:419 arg1 TCR 435:437 NKT TCR 431:437 NKT TCR 431:437 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 5 31 dep complexes 870:878 arg1 contrast 822:829 contrast 822:829 contrast 822:829 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 1 32 theme large 94:98 arg1 family 82:87 The CD1 family 74:87 The CD1 family 74:87 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 1 32 theme large 94:98 arg1 cluster 100:106 a large cluster 92:106 a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells 92:259 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 5 33 theme type 992:995 arg1 interaction 997:1007 a lock-and-key type interaction 977:1007 a lock-and-key type interaction with the lipid antigen 977:1030 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 2 34 theme semi-invariant 399:412 arg1 receptor 421:428 a semi-invariant T-cell receptor 397:428 a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d 397:495 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 34 theme semi-invariant 399:412 arg1 TCR 435:437 NKT TCR 431:437 NKT TCR 431:437 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 35 theme studied 271:277 arg1 group 279:283 The most studied group 262:283 The most studied group of T cells that interact with lipid antigens 262:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 35 theme studied 271:277 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 35 theme studied 271:277 arg1 cells 357:361 natural killer T (NKT) cells 334:361 natural killer T (NKT) cells 334:361 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 36 theme T 288:288 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 1 37 theme distinct 200:207 arg1 antigens 221:228 distinct lipid-based antigens 200:228 distinct lipid-based antigens that are recognized by T cells 200:259 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 0 38 theme NKT 53:55 arg1 receptor 64:71 the semi-invariant NKT T-cell receptor 34:71 the semi-invariant NKT T-cell receptor 34:71 CD1d-lipid-antigen recognition by the semi-invariant NKT T-cell receptor. 17581592 4 39 theme human 678:682 arg1 TCR 688:690 a human NKT TCR 676:690 a human NKT TCR 676:690 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 6 40 theme CD1d-antigen 1295:1306 arg1 complexes 1308:1316 other CD1d-antigen complexes 1289:1316 other CD1d-antigen complexes 1289:1316 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 6 41 theme conserved 1103:1111 arg1 alpha-chain 1121:1131 the highly conserved NKT TCR alpha-chain 1092:1131 the highly conserved NKT TCR alpha-chain 1092:1131 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 4 42 theme CD1d-restricted 791:805 arg1 glycolipid 807:816 the archetypal CD1d-restricted glycolipid 776:816 the archetypal CD1d-restricted glycolipid 776:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 4 42 theme CD1d-restricted 791:805 arg1 alpha-galactosylceramide 750:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 7 43 theme T-cell 1368:1373 arg1 receptor 1375:1382 a T-cell receptor 1366:1382 a T-cell receptor 1366:1382 These findings provide direct insight into how a T-cell receptor recognizes a lipid-antigen-presenting molecule of the immune system. 17581592 1 44 theme lipid-based 209:219 arg1 antigens 221:228 distinct lipid-based antigens 200:228 distinct lipid-based antigens that are recognized by T cells 200:259 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 0 45 theme semi-invariant 38:51 arg1 receptor 64:71 the semi-invariant NKT T-cell receptor 34:71 the semi-invariant NKT T-cell receptor 34:71 CD1d-lipid-antigen recognition by the semi-invariant NKT T-cell receptor. 17581592 6 46 theme other 1289:1293 arg1 complexes 1308:1316 other CD1d-antigen complexes 1289:1316 other CD1d-antigen complexes 1289:1316 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 3 47 theme microbial 579:587 arg1 immunity 589:596 microbial immunity 579:596 microbial immunity 579:596 NKT-cell-mediated recognition of the CD1d-antigen complex has been implicated in microbial immunity, tumour immunity, autoimmunity and allergy. 17581592 2 48 theme family 477:482 arg1 member 484:489 the CD1 family member 469:489 the CD1 family member 469:489 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 48 theme family 477:482 arg1 CD1d 492:495 CD1d 492:495 CD1d 492:495 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 5 49 theme NKT 885:887 arg1 TCR 889:891 the NKT TCR 881:891 the NKT TCR 881:891 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 5 50 theme cleft 956:960 arg1 end 932:934 the extreme end 920:934 the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen 920:1030 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 4 51 theme NKT 684:686 arg1 TCR 688:690 a human NKT TCR 676:690 a human NKT TCR 676:690 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 0 52 theme T-cell 57:62 arg1 receptor 64:71 the semi-invariant NKT T-cell receptor 34:71 the semi-invariant NKT T-cell receptor 34:71 CD1d-lipid-antigen recognition by the semi-invariant NKT T-cell receptor. 17581592 5 53 theme lipid 1018:1022 arg1 antigen 1024:1030 the lipid antigen 1014:1030 the lipid antigen 1014:1030 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 1 54 theme complex 153:159 arg1 molecules 180:188 non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules 111:188 molecules 180:188 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 1 55 theme non-polymorphic 111:125 arg1 molecules 180:188 non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules 111:188 molecules 180:188 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 4 56 with complex 695:701 arg1 CD1d 708:711 CD1d 708:711 CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid 708:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 3 57 theme tumour 599:604 arg1 immunity 606:613 tumour immunity 599:613 tumour immunity 599:613 NKT-cell-mediated recognition of the CD1d-antigen complex has been implicated in microbial immunity, tumour immunity, autoimmunity and allergy. 17581592 4 58 theme archetypal 780:789 arg1 glycolipid 807:816 the archetypal CD1d-restricted glycolipid 776:816 the archetypal CD1d-restricted glycolipid 776:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 4 58 theme archetypal 780:789 arg1 alpha-galactosylceramide 750:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 2 59 theme lipid 315:319 arg1 antigens 321:328 lipid antigens 315:328 lipid antigens 315:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 60 theme cells 290:294 arg1 group 279:283 The most studied group 262:283 The most studied group of T cells that interact with lipid antigens 262:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 60 theme cells 290:294 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 60 theme cells 290:294 arg1 cells 357:361 natural killer T (NKT) cells 334:361 natural killer T (NKT) cells 334:361 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 4 61 theme NKT-cell 733:740 arg1 glycolipid 807:816 the archetypal CD1d-restricted glycolipid 776:816 the archetypal CD1d-restricted glycolipid 776:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 4 61 theme NKT-cell 733:740 arg1 alpha-galactosylceramide 750:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 4 62 theme agonist 742:748 arg1 glycolipid 807:816 the archetypal CD1d-restricted glycolipid 776:816 the archetypal CD1d-restricted glycolipid 776:816 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 4 62 theme agonist 742:748 arg1 alpha-galactosylceramide 750:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 the potent NKT-cell agonist alpha-galactosylceramide 722:773 Here we describe the structure of a human NKT TCR in complex with CD1d bound to the potent NKT-cell agonist alpha-galactosylceramide, the archetypal CD1d-restricted glycolipid. 17581592 1 63 theme major 128:132 arg1 molecules 180:188 non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules 111:188 molecules 180:188 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 2 64 theme CD1 473:475 arg1 member 484:489 the CD1 family member 469:489 the CD1 family member 469:489 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 64 theme CD1 473:475 arg1 CD1d 492:495 CD1d 492:495 CD1d 492:495 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 5 65 theme T-cell 834:839 arg1 complexes 870:878 T-cell receptor-peptide-antigen-MHC complexes 834:878 T-cell receptor-peptide-antigen-MHC complexes 834:878 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 1 66 theme histocompatibility 134:151 arg1 molecules 180:188 non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules 111:188 molecules 180:188 The CD1 family is a large cluster of non-polymorphic, major histocompatibility complex (MHC) class-I-like molecules that bind distinct lipid-based antigens that are recognized by T cells. 17581592 6 67 theme TCR 1117:1119 arg1 alpha-chain 1121:1131 the highly conserved NKT TCR alpha-chain 1092:1131 the highly conserved NKT TCR alpha-chain 1092:1131 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 5 68 theme lock-and-key 979:990 arg1 interaction 997:1007 a lock-and-key type interaction 977:1007 a lock-and-key type interaction with the lipid antigen 977:1030 In contrast to T-cell receptor-peptide-antigen-MHC complexes, the NKT TCR docked parallel to, and at the extreme end of the CD1d-binding cleft, which enables a lock-and-key type interaction with the lipid antigen. 17581592 2 69 theme T 349:349 arg1 cells 357:361 natural killer T (NKT) cells 334:361 natural killer T (NKT) cells 334:361 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 69 theme T 349:349 arg1 group 279:283 The most studied group 262:283 The most studied group of T cells that interact with lipid antigens 262:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 2 69 theme T 349:349 arg1 cells 290:294 T cells 288:294 T cells that interact with lipid antigens 288:328 The most studied group of T cells that interact with lipid antigens are natural killer T (NKT) cells, which characteristically express a semi-invariant T-cell receptor (NKT TCR) that specifically recognizes the CD1 family member, CD1d. 17581592 6 70 theme TCR 1245:1247 arg1 beta-chain 1249:1258 the NKT TCR beta-chain 1237:1258 the NKT TCR beta-chain 1237:1258 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 17581592 6 71 from impact 1264:1269 arg1 recognition 1274:1284 recognition 1274:1284 recognition of other CD1d-antigen complexes 1274:1316 The structure provides a basis for the interaction between the highly conserved NKT TCR alpha-chain and the CD1d-antigen complex that is typified in innate immunity, and also indicates how variability of the NKT TCR beta-chain can impact on recognition of other CD1d-antigen complexes. 21244856 4 0 theme serine 684:689 arg1 acylation 671:679 O-linked acylation 662:679 O-linked acylation of serine 662:689 We revealed that O-linked acylation of serine is required for the subsequent S-palmitoylation of cysteine. 21244856 1 1 theme cell-fate 222:230 arg1 decisions 232:240 cell-fate decisions 222:240 cell-fate decisions in developing and adult tissues 222:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 7 2 from prominent 1095:1103 arg1 case 1026:1029 the case 1022:1029 the case of Wnt3a 1022:1038 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 2 3 theme family 316:321 arg1 members 323:329 all 19 mammalian Wnt family members 295:329 all 19 mammalian Wnt family members 295:329 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 6 4 theme signalling 998:1007 arg1 activity 1009:1016 the signalling activity 994:1016 the signalling activity 994:1016 Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. 21244856 7 5 theme various 1185:1191 arg1 contexts 1202:1209 various cellular contexts 1185:1209 various cellular contexts 1185:1209 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 4 6 link O-linked 662:669 arg1 acylation 671:679 O-linked acylation 662:679 O-linked acylation of serine 662:689 We revealed that O-linked acylation of serine is required for the subsequent S-palmitoylation of cysteine. 21244856 6 7 theme S-palmitoyl-deficient 958:978 arg1 form 980:983 the S-palmitoyl-deficient form 954:983 the S-palmitoyl-deficient form 954:983 Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. 21244856 5 8 theme crucial 791:797 arg1 residue 806:812 the crucial serine residue 787:812 the crucial serine residue 787:812 As such, mutant proteins that lack the crucial serine residue are not lipidated. 21244856 8 9 theme Wnt1 1268:1271 arg1 competency 1249:1258 the signalling competency 1234:1258 the signalling competency of both Wnt1 and Wnt3a 1234:1281 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 8 9 theme Wnt1 1268:1271 arg1 related 1286:1292 related 1286:1292 related 1286:1292 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 8 10 theme Wnt3a 1277:1281 arg1 competency 1249:1258 the signalling competency 1234:1258 the signalling competency of both Wnt1 and Wnt3a 1234:1281 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 8 10 theme Wnt3a 1277:1281 arg1 related 1286:1292 related 1286:1292 related 1286:1292 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 2 11 theme asparagines 487:497 arg1 N-glycosylation 459:473 the N-glycosylation 455:473 the N-glycosylation of multiple asparagines 455:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 11 theme asparagines 487:497 arg1 types 343:347 two types 339:347 two types of post-translational modification 339:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 11 theme asparagines 487:497 arg1 attachment 398:407 the covalent attachment 385:407 the covalent attachment of fatty acids at two distinct positions 385:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 8 12 theme signalling 1238:1247 arg1 competency 1249:1258 the signalling competency 1234:1258 the signalling competency of both Wnt1 and Wnt3a 1234:1281 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 8 12 theme signalling 1238:1247 arg1 related 1286:1292 related 1286:1292 related 1286:1292 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 2 13 theme modification 371:382 arg1 N-glycosylation 459:473 the N-glycosylation 455:473 the N-glycosylation of multiple asparagines 455:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 13 theme modification 371:382 arg1 types 343:347 two types 339:347 two types of post-translational modification 339:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 13 theme modification 371:382 arg1 attachment 398:407 the covalent attachment 385:407 the covalent attachment of fatty acids at two distinct positions 385:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 14 theme multiple 478:485 arg1 asparagines 487:497 multiple asparagines 478:497 multiple asparagines 478:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 15 contain contain 331:337 arg2 attachment 398:407 the covalent attachment 385:407 the covalent attachment of fatty acids at two distinct positions 385:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 15 contain contain 331:337 arg1 members 323:329 all 19 mammalian Wnt family members 295:329 all 19 mammalian Wnt family members 295:329 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 15 contain contain 331:337 arg2 N-glycosylation 459:473 the N-glycosylation 455:473 the N-glycosylation of multiple asparagines 455:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 15 contain contain 331:337 arg2 types 343:347 two types 339:347 two types of post-translational modification 339:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 7 16 theme S-linked 1131:1138 arg1 palmitate 1140:1148 S-linked palmitate 1131:1148 S-linked palmitate 1131:1148 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 2 17 theme acids 418:422 arg1 N-glycosylation 459:473 the N-glycosylation 455:473 the N-glycosylation of multiple asparagines 455:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 17 theme acids 418:422 arg1 types 343:347 two types 339:347 two types of post-translational modification 339:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 17 theme acids 418:422 arg1 attachment 398:407 the covalent attachment 385:407 the covalent attachment of fatty acids at two distinct positions 385:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 18 theme post-translational 352:369 arg1 modification 371:382 post-translational modification 352:382 post-translational modification 352:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 0 19 theme acid 6:9 arg1 modification 11:22 Fatty acid modification 0:22 Fatty acid modification of Wnt1 and Wnt3a at serine 0:50 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 2 20 from positions 440:448 arg1 N-glycosylation 459:473 the N-glycosylation 455:473 the N-glycosylation of multiple asparagines 455:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 20 from positions 440:448 arg1 acids 418:422 fatty acids 412:422 fatty acids at two distinct positions 412:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 20 from positions 440:448 arg1 types 343:347 two types 339:347 two types of post-translational modification 339:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 20 from positions 440:448 arg1 attachment 398:407 the covalent attachment 385:407 the covalent attachment of fatty acids at two distinct positions 385:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 7 21 theme attached 1071:1078 arg1 acyls 1080:1084 the attached acyls 1067:1084 the attached acyls 1067:1084 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 2 22 gly N-glycosylation 459:473 arg1 acids 418:422 fatty acids 412:422 fatty acids at two distinct positions 412:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 22 gly N-glycosylation 459:473 arg1 asparagines 487:497 multiple asparagines 478:497 multiple asparagines 478:497 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 22 gly N-glycosylation 459:473 arg1 modification 371:382 post-translational modification 352:382 post-translational modification 352:382 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 1 23 theme proteins 150:157 arg1 group 164:168 a group 162:168 a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 162:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 23 theme proteins 150:157 arg1 family 140:145 The Wnt family 132:145 The Wnt family of proteins 132:157 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 23 theme proteins 150:157 arg1 molecules 198:206 extracellular signalling molecules 173:206 extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 173:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 0 24 theme Fatty 0:4 arg1 modification 11:22 Fatty acid modification 0:22 Fatty acid modification of Wnt1 and Wnt3a at serine 0:50 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 7 25 theme acyls 1080:1084 arg1 duality 1056:1062 the functional duality 1041:1062 the functional duality of the attached acyls 1041:1084 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 7 25 theme acyls 1080:1084 arg1 prominent 1095:1103 prominent 1095:1103 prominent 1095:1103 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 4 26 theme subsequent 711:720 arg1 S-palmitoylation 722:737 the subsequent S-palmitoylation 707:737 the subsequent S-palmitoylation of cysteine 707:749 We revealed that O-linked acylation of serine is required for the subsequent S-palmitoylation of cysteine. 21244856 2 27 theme fatty 412:416 arg1 acids 418:422 fatty acids 412:422 fatty acids at two distinct positions 412:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 1 28 theme adult 260:264 arg1 tissues 266:272 adult tissues 260:272 adult tissues 260:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 2 29 from N-glycosylation 459:473 arg1 positions 440:448 two distinct positions 427:448 two distinct positions 427:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 4 30 theme cysteine 742:749 arg1 S-palmitoylation 722:737 the subsequent S-palmitoylation 707:737 the subsequent S-palmitoylation of cysteine 707:749 We revealed that O-linked acylation of serine is required for the subsequent S-palmitoylation of cysteine. 21244856 8 31 theme extracellular 1333:1345 arg1 matrix 1347:1352 the extracellular matrix 1329:1352 the extracellular matrix 1329:1352 Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix. 21244856 7 32 theme functional 1045:1054 arg1 duality 1056:1062 the functional duality 1041:1062 the functional duality of the attached acyls 1041:1084 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 7 32 theme functional 1045:1054 arg1 prominent 1095:1103 prominent 1095:1103 prominent 1095:1103 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 3 33 theme Wnt3a 630:634 arg1 movement 579:586 extracellular movement 565:586 extracellular movement 565:586 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 3 33 theme Wnt3a 630:634 arg1 activity 603:610 signalling activity 592:610 signalling activity 592:610 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 3 33 theme Wnt3a 630:634 arg1 secretion 554:562 secretion 554:562 secretion 554:562 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 1 34 theme Wnt 136:138 arg1 group 164:168 a group 162:168 a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 162:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 34 theme Wnt 136:138 arg1 family 140:145 The Wnt family 132:145 The Wnt family of proteins 132:157 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 34 theme Wnt 136:138 arg1 molecules 198:206 extracellular signalling molecules 173:206 extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 173:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 3 35 dep secretion 554:562 arg1 the 550:552 the 550:552 the 550:552 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 2 36 theme covalent 389:396 arg1 attachment 398:407 the covalent attachment 385:407 the covalent attachment of fatty acids at two distinct positions 385:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 5 37 theme mutant 761:766 arg1 proteins 768:775 mutant proteins 761:775 mutant proteins that lack the crucial serine residue 761:812 As such, mutant proteins that lack the crucial serine residue are not lipidated. 21244856 6 38 theme Wnt1 877:880 arg1 double-acylation 857:872 double-acylation 857:872 double-acylation of Wnt1 857:880 Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. 21244856 0 39 theme Wnt 116:118 arg1 signalling 120:129 Wnt signalling 116:129 Wnt signalling 116:129 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 7 40 from signalling 1171:1180 arg1 contexts 1202:1209 various cellular contexts 1185:1209 various cellular contexts 1185:1209 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 0 41 theme Wnt1 27:30 arg1 modification 11:22 Fatty acid modification 0:22 Fatty acid modification of Wnt1 and Wnt3a at serine 0:50 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 0 42 mod modification 11:22 arg3 acid 6:9 Fatty acid modification 0:22 Fatty acid modification of Wnt1 and Wnt3a at serine 0:50 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 0 42 mod modification 11:22 arg1 Wnt3a 36:40 Wnt3a 36:40 Wnt3a 36:40 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 0 42 mod modification 11:22 arg1 Wnt1 27:30 Wnt1 27:30 Wnt1 27:30 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 1 43 from decisions 232:240 arg1 tissues 266:272 adult tissues 260:272 adult tissues 260:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 43 from decisions 232:240 arg1 developing 245:254 developing 245:254 developing 245:254 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 3 44 theme Wnt1 621:624 arg1 movement 579:586 extracellular movement 565:586 extracellular movement 565:586 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 3 44 theme Wnt1 621:624 arg1 activity 603:610 signalling activity 592:610 signalling activity 592:610 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 3 44 theme Wnt1 621:624 arg1 secretion 554:562 secretion 554:562 secretion 554:562 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 5 45 theme serine 799:804 arg1 residue 806:812 the crucial serine residue 787:812 the crucial serine residue 787:812 As such, mutant proteins that lack the crucial serine residue are not lipidated. 21244856 1 46 theme extracellular 173:185 arg1 molecules 198:206 extracellular signalling molecules 173:206 extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 173:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 2 47 theme distinct 431:438 arg1 positions 440:448 two distinct positions 427:448 two distinct positions 427:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 6 48 theme Xenopus 938:944 arg1 embryos 946:952 Xenopus embryos 938:952 Xenopus embryos 938:952 Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. 21244856 0 49 from serine 45:50 arg1 modification 11:22 Fatty acid modification 0:22 Fatty acid modification of Wnt1 and Wnt3a at serine 0:50 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 7 50 link S-linked 1131:1138 arg1 palmitate 1140:1148 S-linked palmitate 1131:1148 S-linked palmitate 1131:1148 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 2 51 from types 343:347 arg1 positions 440:448 two distinct positions 427:448 two distinct positions 427:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 2 52 theme Wnt 312:314 arg1 members 323:329 all 19 mammalian Wnt family members 295:329 all 19 mammalian Wnt family members 295:329 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 0 53 theme Wnt3a 36:40 arg1 modification 11:22 Fatty acid modification 0:22 Fatty acid modification of Wnt1 and Wnt3a at serine 0:50 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 3 54 theme extracellular 565:577 arg1 movement 579:586 extracellular movement 565:586 extracellular movement 565:586 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 2 55 theme mammalian 302:310 arg1 members 323:329 all 19 mammalian Wnt family members 295:329 all 19 mammalian Wnt family members 295:329 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 0 56 from cysteine 86:93 arg1 prerequisite 55:66 prerequisite 55:66 prerequisite for lipidation 55:81 Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling. 21244856 1 57 theme signalling 187:196 arg1 molecules 198:206 extracellular signalling molecules 173:206 extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 173:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 7 58 theme Wnt3a 1034:1038 arg1 case 1026:1029 the case 1022:1029 the case of Wnt3a 1022:1038 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 3 59 theme mouse 615:619 arg1 Wnt1 621:624 mouse Wnt1 615:624 mouse Wnt1 615:624 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 4 60 theme O-linked 662:669 arg1 acylation 671:679 O-linked acylation 662:679 O-linked acylation of serine 662:689 We revealed that O-linked acylation of serine is required for the subsequent S-palmitoylation of cysteine. 21244856 1 61 theme molecules 198:206 arg1 group 164:168 a group 162:168 a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 162:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 61 theme molecules 198:206 arg1 family 140:145 The Wnt family 132:145 The Wnt family of proteins 132:157 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 1 61 theme molecules 198:206 arg1 molecules 198:206 extracellular signalling molecules 173:206 extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues 173:272 The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. 21244856 6 62 from signalling 904:913 arg1 cells 928:932 mammalian cells 918:932 mammalian cells 918:932 Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. 21244856 7 63 from case 1026:1029 arg1 duality 1056:1062 the functional duality 1041:1062 the functional duality of the attached acyls 1041:1084 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 7 63 from case 1026:1029 arg1 prominent 1095:1103 prominent 1095:1103 prominent 1095:1103 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 3 64 theme signalling 592:601 arg1 activity 603:610 signalling activity 592:610 signalling activity 592:610 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 7 65 theme signalling 1171:1180 arg1 ligand 1116:1121 the ligand 1112:1121 the ligand lacking S-linked palmitate 1112:1148 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 7 65 theme signalling 1171:1180 arg1 capable 1160:1166 capable 1160:1166 capable 1160:1166 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 21244856 6 66 theme mammalian 918:926 arg1 cells 928:932 mammalian cells 918:932 mammalian cells 918:932 Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. 21244856 2 67 from attachment 398:407 arg1 positions 440:448 two distinct positions 427:448 two distinct positions 427:448 It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. 21244856 3 68 dep Wnt1 621:624 arg1 ligands 636:642 ligands 636:642 ligands 636:642 We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. 21244856 7 69 theme cellular 1193:1200 arg1 contexts 1202:1209 various cellular contexts 1185:1209 various cellular contexts 1185:1209 In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. 23756652 5 0 theme FU-CRD2 716:722 arg1 RSPO1 735:739 RSPO1 735:739 RSPO1 735:739 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 5 0 theme FU-CRD2 716:722 arg1 domains 724:730 Both the FU-CRD1 and FU-CRD2 domains 695:730 domains 724:730 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 5 1 theme binding 777:783 arg1 assays 798:803 binding and cellular assays 777:803 assays 798:803 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 1 2 theme pleiotropic 133:143 arg1 functions 145:153 pleiotropic functions 133:153 pleiotropic functions 133:153 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 5 3 theme biological 848:857 arg1 activities 859:868 its biological activities 844:868 its biological activities 844:868 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 3 4 theme N-terminal 466:475 arg1 RSPO1-2F 487:494 RSPO1-2F 487:494 RSPO1-2F 487:494 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 4 theme N-terminal 466:475 arg1 fragment 477:484 the RSPO1 N-terminal fragment 456:484 the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs) 456:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 6 5 theme signaling 1008:1016 arg1 difference 1018:1027 the signaling difference 1004:1027 the signaling difference between the LGR4/5/6 receptors and other members in the LGR family 1004:1094 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 3 6 theme cysteine-rich 532:544 arg1 FU-CRDs 555:561 FU-CRDs 555:561 FU-CRDs 555:561 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 6 theme cysteine-rich 532:544 arg1 domains 546:552 two adjacent furin-like cysteine-rich domains 508:552 two adjacent furin-like cysteine-rich domains (FU-CRDs) 508:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 2 7 theme receptor 296:303 arg1 LGR4 308:311 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 7 theme receptor 296:303 arg1 receptors 354:362 receptors 354:362 receptors for RSPOs 354:372 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 7 theme receptor 296:303 arg1 LGR6 325:328 LGR6 325:328 LGR6 325:328 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 7 theme receptor 296:303 arg1 LGR5 315:318 LGR5 315:318 LGR5 315:318 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 6 8 from receptors 1050:1058 arg1 family 1089:1094 the LGR family 1081:1094 the LGR family 1081:1094 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 1 9 theme R-spondin 70:78 arg1 family 87:92 The R-spondin (RSPO) family 66:92 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) 66:127 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 4 10 theme concave 638:644 arg1 surface 646:652 its concave surface 634:652 its concave surface close to the N termini to bind RSPO1-2F 634:692 The LGR4-ECD adopts the anticipated TLR horseshoe structure and uses its concave surface close to the N termini to bind RSPO1-2F. 23756652 1 11 contain has 129:131 arg2 functions 145:153 pleiotropic functions 133:153 pleiotropic functions 133:153 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 1 11 contain has 129:131 arg1 family 87:92 The R-spondin (RSPO) family 66:92 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) 66:127 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 2 12 theme repeat-containing 260:276 arg1 LGR4 308:311 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 12 theme repeat-containing 260:276 arg1 receptors 354:362 receptors 354:362 receptors for RSPOs 354:372 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 12 theme repeat-containing 260:276 arg1 LGR6 325:328 LGR6 325:328 LGR6 325:328 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 12 theme repeat-containing 260:276 arg1 LGR5 315:318 LGR5 315:318 LGR5 315:318 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 6 13 theme other 1064:1068 arg1 members 1070:1076 other members 1064:1076 other members in the LGR family 1064:1094 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 5 14 theme critical 816:823 arg1 residues 831:838 critical RSPO1 residues 816:838 critical RSPO1 residues for its biological activities 816:868 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 5 15 theme LGR4 755:758 arg1 interaction 760:770 LGR4 interaction 755:770 LGR4 interaction 755:770 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 1 16 theme RSPO 81:84 arg1 family 87:92 The R-spondin (RSPO) family 66:92 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) 66:127 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 6 17 theme LGR4/5/6 1041:1048 arg1 receptors 1050:1058 the LGR4/5/6 receptors 1037:1058 the LGR4/5/6 receptors 1037:1058 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 3 18 theme complex 394:400 arg1 structure 402:410 the complex structure 390:410 the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs) 390:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 0 19 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for R-spondin recognition by LGR4/5/6 receptors 0:63 Structural basis for R-spondin recognition by LGR4/5/6 receptors. 23756652 1 20 theme stem 174:177 arg1 growth 184:189 stem cell growth 174:189 stem cell growth 174:189 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 0 21 theme R-spondin 21:29 arg1 recognition 31:41 R-spondin recognition 21:41 R-spondin recognition by LGR4/5/6 receptors 21:63 Structural basis for R-spondin recognition by LGR4/5/6 receptors. 23756652 3 22 theme adjacent 512:519 arg1 FU-CRDs 555:561 FU-CRDs 555:561 FU-CRDs 555:561 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 22 theme adjacent 512:519 arg1 domains 546:552 two adjacent furin-like cysteine-rich domains 508:552 two adjacent furin-like cysteine-rich domains (FU-CRDs) 508:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 23 theme domain 438:443 arg1 structure 402:410 the complex structure 390:410 the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs) 390:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 2 24 theme G-protein-coupled 278:294 arg1 LGR4 308:311 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 24 theme G-protein-coupled 278:294 arg1 receptors 354:362 receptors 354:362 receptors for RSPOs 354:372 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 24 theme G-protein-coupled 278:294 arg1 LGR6 325:328 LGR6 325:328 LGR6 325:328 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 24 theme G-protein-coupled 278:294 arg1 LGR5 315:318 LGR5 315:318 LGR5 315:318 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 3 25 theme furin-like 521:530 arg1 FU-CRDs 555:561 FU-CRDs 555:561 FU-CRDs 555:561 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 25 theme furin-like 521:530 arg1 domains 546:552 two adjacent furin-like cysteine-rich domains 508:552 two adjacent furin-like cysteine-rich domains (FU-CRDs) 508:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 4 26 theme anticipated 589:599 arg1 structure 615:623 the anticipated TLR horseshoe structure 585:623 the anticipated TLR horseshoe structure 585:623 The LGR4-ECD adopts the anticipated TLR horseshoe structure and uses its concave surface close to the N termini to bind RSPO1-2F. 23756652 3 27 theme RSPO1 460:464 arg1 RSPO1-2F 487:494 RSPO1-2F 487:494 RSPO1-2F 487:494 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 27 theme RSPO1 460:464 arg1 fragment 477:484 the RSPO1 N-terminal fragment 456:484 the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs) 456:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 4 28 theme N 667:667 arg1 termini 669:675 the N termini 663:675 the N termini to bind RSPO1-2F 663:692 The LGR4-ECD adopts the anticipated TLR horseshoe structure and uses its concave surface close to the N termini to bind RSPO1-2F. 23756652 5 29 theme cellular 789:796 arg1 assays 798:803 binding and cellular assays 777:803 assays 798:803 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 3 30 theme LGR4 419:422 arg1 ECD 446:448 ECD 446:448 ECD 446:448 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 30 theme LGR4 419:422 arg1 domain 438:443 LGR4 extracellular domain 419:443 the LGR4 extracellular domain (ECD) 415:449 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 5 31 theme RSPO1 735:739 arg1 RSPO1 735:739 RSPO1 735:739 RSPO1 735:739 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 5 31 theme RSPO1 735:739 arg1 domains 724:730 Both the FU-CRD1 and FU-CRD2 domains 695:730 domains 724:730 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 5 32 theme RSPO1 825:829 arg1 residues 831:838 critical RSPO1 residues 816:838 critical RSPO1 residues for its biological activities 816:868 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 6 33 theme structural 979:988 arg1 insights 990:997 structural insights 979:997 structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family 979:1094 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 3 34 theme extracellular 424:436 arg1 ECD 446:448 ECD 446:448 ECD 446:448 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 34 theme extracellular 424:436 arg1 domain 438:443 LGR4 extracellular domain 419:443 the LGR4 extracellular domain (ECD) 415:449 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 1 35 theme secreted 97:104 arg1 RSPO1-RSPO4 116:126 RSPO1-RSPO4 116:126 RSPO1-RSPO4 116:126 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 1 35 theme secreted 97:104 arg1 proteins 106:113 secreted proteins 97:113 secreted proteins (RSPO1-RSPO4) 97:127 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 0 36 theme LGR4/5/6 46:53 arg1 receptors 55:63 LGR4/5/6 receptors 46:63 LGR4/5/6 receptors 46:63 Structural basis for R-spondin recognition by LGR4/5/6 receptors. 23756652 6 37 theme molecular 894:902 arg1 mechanism 904:912 the molecular mechanism 890:912 the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs 890:960 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 3 38 with structure 402:410 arg1 RSPO1-2F 487:494 RSPO1-2F 487:494 RSPO1-2F 487:494 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 38 with structure 402:410 arg1 fragment 477:484 the RSPO1 N-terminal fragment 456:484 the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs) 456:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 1 39 theme proteins 106:113 arg1 family 87:92 The R-spondin (RSPO) family 66:92 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) 66:127 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 3 40 contain containing 497:506 arg2 domains 546:552 two adjacent furin-like cysteine-rich domains 508:552 two adjacent furin-like cysteine-rich domains (FU-CRDs) 508:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 40 contain containing 497:506 arg1 RSPO1-2F 487:494 RSPO1-2F 487:494 RSPO1-2F 487:494 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 40 contain containing 497:506 arg1 fragment 477:484 the RSPO1 N-terminal fragment 456:484 the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs) 456:562 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 3 40 contain containing 497:506 arg2 FU-CRDs 555:561 FU-CRDs 555:561 FU-CRDs 555:561 Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). 23756652 2 41 theme leucine-rich 247:258 arg1 LGR4 308:311 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) 247:312 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 41 theme leucine-rich 247:258 arg1 receptors 354:362 receptors 354:362 receptors for RSPOs 354:372 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 41 theme leucine-rich 247:258 arg1 LGR6 325:328 LGR6 325:328 LGR6 325:328 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 2 41 theme leucine-rich 247:258 arg1 LGR5 315:318 LGR5 315:318 LGR5 315:318 Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. 23756652 6 42 from members 1070:1076 arg1 family 1089:1094 the LGR family 1081:1094 the LGR family 1081:1094 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 6 43 theme LGR 1085:1087 arg1 family 1089:1094 the LGR family 1081:1094 the LGR family 1081:1094 Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family. 23756652 1 44 theme Wnt 213:215 arg1 activation 225:234 Wnt pathway activation 213:234 Wnt pathway activation 213:234 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 5 45 theme FU-CRD1 704:710 arg1 RSPO1 735:739 RSPO1 735:739 RSPO1 735:739 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 5 45 theme FU-CRD1 704:710 arg1 domains 724:730 Both the FU-CRD1 and FU-CRD2 domains 695:730 domains 724:730 Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. 23756652 4 46 theme horseshoe 605:613 arg1 structure 615:623 the anticipated TLR horseshoe structure 585:623 the anticipated TLR horseshoe structure 585:623 The LGR4-ECD adopts the anticipated TLR horseshoe structure and uses its concave surface close to the N termini to bind RSPO1-2F. 23756652 1 47 theme cell 179:182 arg1 growth 184:189 stem cell growth 174:189 stem cell growth 174:189 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 1 48 theme pathway 217:223 arg1 activation 225:234 Wnt pathway activation 213:234 Wnt pathway activation 213:234 The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. 23756652 4 49 theme TLR 601:603 arg1 structure 615:623 the anticipated TLR horseshoe structure 585:623 the anticipated TLR horseshoe structure 585:623 The LGR4-ECD adopts the anticipated TLR horseshoe structure and uses its concave surface close to the N termini to bind RSPO1-2F. 28753425 5 0 theme vasopressin-2 944:956 arg1 tail 967:970 the phosphorylated vasopressin-2 receptor tail 925:970 the phosphorylated vasopressin-2 receptor tail 925:970 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 1 1 theme G 273:273 arg1 pathways 295:302 G protein-independent pathways 273:302 G protein-independent pathways 273:302 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 5 2 theme electrostatic 812:824 arg1 interactions 826:837 electrostatic interactions 812:837 electrostatic interactions with three positively charged pockets in arrestin 812:887 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 2 3 theme C-terminal 395:404 arg1 tail 406:409 the receptor's C-terminal tail 380:409 the receptor's C-terminal tail 380:409 High-affinity arrestin binding requires receptor phosphorylation, often at the receptor's C-terminal tail. 28753425 1 4 theme protein-independent 275:293 arg1 pathways 295:302 G protein-independent pathways 273:302 G protein-independent pathways 273:302 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 3 5 theme intermolecular 587:600 arg1 β sheet 602:608 an extended intermolecular β sheet 575:608 an extended intermolecular β sheet with the N-terminal β strands of arrestin 575:650 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 5 6 from pockets 869:875 arg1 arrestin 880:887 arrestin 880:887 arrestin 880:887 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 1 7 from signaling 149:157 arg1 part 162:165 part 162:165 part 162:165 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 6 8 dep phosphorylation 1052:1066 arg1 codes 1068:1072 codes 1068:1072 codes 1068:1072 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 6 9 theme common 1090:1095 arg1 set 1045:1047 a set 1043:1047 a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs 1043:1169 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 6 9 theme common 1090:1095 arg1 mechanism 1097:1105 a common mechanism 1088:1105 a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs 1088:1169 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 6 9 theme common 1090:1095 arg1 phosphorylation 1052:1066 phosphorylation codes 1052:1072 phosphorylation codes 1052:1072 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 2 10 theme arrestin 319:326 arg1 binding 328:334 High-affinity arrestin binding 305:334 High-affinity arrestin binding 305:334 High-affinity arrestin binding requires receptor phosphorylation, often at the receptor's C-terminal tail. 28753425 4 11 theme tail 706:709 arg1 residues 711:718 rhodopsin C-terminal tail residues T336 and S338 685:732 rhodopsin C-terminal tail residues T336 and S338 685:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 11 theme tail 706:709 arg1 S338 729:732 S338 729:732 S338 729:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 11 theme tail 706:709 arg1 T336 720:723 T336 720:723 T336 720:723 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 5 12 theme receptor 958:965 arg1 tail 967:970 the phosphorylated vasopressin-2 receptor tail 925:970 the phosphorylated vasopressin-2 receptor tail 925:970 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 3 13 theme electron 442:449 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 2 14 theme High-affinity 305:317 arg1 binding 328:334 High-affinity arrestin binding 305:334 High-affinity arrestin binding 305:334 High-affinity arrestin binding requires receptor phosphorylation, often at the receptor's C-terminal tail. 28753425 6 15 theme arrestins 1152:1160 arg1 recruitment 1137:1147 phosphorylation-dependent recruitment 1111:1147 phosphorylation-dependent recruitment of arrestins by GPCRs 1111:1169 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 3 16 theme crystal 464:470 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 6 17 theme phosphorylation-dependent 1111:1135 arg1 recruitment 1137:1147 phosphorylation-dependent recruitment 1111:1147 phosphorylation-dependent recruitment of arrestins by GPCRs 1111:1169 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 5 18 theme charged 861:867 arg1 pockets 869:875 three positively charged pockets 844:875 three positively charged pockets in arrestin 844:887 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 1 19 theme protein-coupled 99:113 arg1 GPCRs 126:130 GPCRs 126:130 GPCRs 126:130 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 1 19 theme protein-coupled 99:113 arg1 receptors 115:123 G protein-coupled receptors 97:123 G protein-coupled receptors (GPCRs) 97:131 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 3 20 theme XFEL 458:461 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 3 21 with β sheet 602:608 arg1 strands 632:638 the N-terminal β strands 615:638 the N-terminal β strands of arrestin 615:650 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 3 22 theme extended 578:585 arg1 β sheet 602:608 an extended intermolecular β sheet 575:608 an extended intermolecular β sheet with the N-terminal β strands of arrestin 575:650 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 0 23 theme Phosphorylation 18:32 arg1 Identification 0:13 Identification 0:13 Identification of Phosphorylation 0:32 Identification of Phosphorylation Codes for Arrestin Recruitment by G Protein-Coupled Receptors. 28753425 2 24 theme receptor 345:352 arg1 phosphorylation 354:368 receptor phosphorylation 345:368 receptor phosphorylation 345:368 High-affinity arrestin binding requires receptor phosphorylation, often at the receptor's C-terminal tail. 28753425 3 25 theme rhodopsin-arrestin 489:506 arg1 complex 508:514 the rhodopsin-arrestin complex 485:514 the rhodopsin-arrestin complex 485:514 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 0 26 theme Arrestin 44:51 arg1 Recruitment 53:63 Arrestin Recruitment 44:63 Arrestin Recruitment by G Protein-Coupled Receptors 44:94 Identification of Phosphorylation Codes for Arrestin Recruitment by G Protein-Coupled Receptors. 28753425 3 27 theme β 630:630 arg1 strands 632:638 the N-terminal β strands 615:638 the N-terminal β strands of arrestin 615:650 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 4 28 theme rhodopsin 685:693 arg1 residues 711:718 rhodopsin C-terminal tail residues T336 and S338 685:732 rhodopsin C-terminal tail residues T336 and S338 685:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 28 theme rhodopsin 685:693 arg1 S338 729:732 S338 729:732 S338 729:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 28 theme rhodopsin 685:693 arg1 T336 720:723 T336 720:723 T336 720:723 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 3 29 theme laser 451:455 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 5 30 theme interactions 826:837 arg1 network 801:807 an extensive network 788:807 an extensive network of electrostatic interactions with three positively charged pockets in arrestin 788:887 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 5 31 with interactions 826:837 arg1 pockets 869:875 three positively charged pockets 844:875 three positively charged pockets in arrestin 844:887 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 1 32 theme receptor 226:233 arg1 internalization 235:249 receptor internalization 226:249 receptor internalization 226:249 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 3 33 theme X-ray 431:435 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 5 34 theme tail 967:970 arg1 binding 914:920 binding 914:920 binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1 914:986 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 3 35 theme free 437:440 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 3 36 theme N-terminal 619:628 arg1 strands 632:638 the N-terminal β strands 615:638 the N-terminal β strands of arrestin 615:650 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 3 37 theme phosphorylated 530:543 arg1 terminus 547:554 the phosphorylated C terminus 526:554 the phosphorylated C terminus of rhodopsin 526:567 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 1 38 with interaction 175:185 arg1 arrestins 192:200 arrestins 192:200 arrestins 192:200 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 4 39 located detected 673:680 arg2 Phosphorylation 653:667 Phosphorylation 653:667 Phosphorylation 653:667 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 39 located detected 673:680 arg1 residues 711:718 rhodopsin C-terminal tail residues T336 and S338 685:732 rhodopsin C-terminal tail residues T336 and S338 685:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 39 located detected 673:680 arg1 S338 729:732 S338 729:732 S338 729:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 39 located detected 673:680 arg1 T336 720:723 T336 720:723 T336 720:723 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 3 40 theme complex 508:514 arg1 structure 472:480 an X-ray free electron laser (XFEL) crystal structure 428:480 an X-ray free electron laser (XFEL) crystal structure 428:480 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 3 41 theme rhodopsin 559:567 arg1 terminus 547:554 the phosphorylated C terminus 526:554 the phosphorylated C terminus of rhodopsin 526:567 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 4 42 dep residues 711:718 arg1 residues 711:718 rhodopsin C-terminal tail residues T336 and S338 685:732 rhodopsin C-terminal tail residues T336 and S338 685:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 42 dep residues 711:718 arg1 S338 729:732 S338 729:732 S338 729:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 42 dep residues 711:718 arg1 T336 720:723 T336 720:723 T336 720:723 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 1 43 theme diverse 141:147 arg1 signaling 149:157 diverse signaling 141:157 diverse signaling in part 141:165 G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. 28753425 0 44 theme Protein-Coupled 70:84 arg1 Receptors 86:94 G Protein-Coupled Receptors 68:94 G Protein-Coupled Receptors 68:94 Identification of Phosphorylation Codes for Arrestin Recruitment by G Protein-Coupled Receptors. 28753425 3 45 theme C 545:545 arg1 terminus 547:554 the phosphorylated C terminus 526:554 the phosphorylated C terminus of rhodopsin 526:567 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 4 46 theme C-terminal 695:704 arg1 residues 711:718 rhodopsin C-terminal tail residues T336 and S338 685:732 rhodopsin C-terminal tail residues T336 and S338 685:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 46 theme C-terminal 695:704 arg1 S338 729:732 S338 729:732 S338 729:732 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 4 46 theme C-terminal 695:704 arg1 T336 720:723 T336 720:723 T336 720:723 Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. 28753425 5 47 theme extensive 791:799 arg1 network 801:807 an extensive network 788:807 an extensive network of electrostatic interactions with three positively charged pockets in arrestin 788:887 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 3 48 theme arrestin 643:650 arg1 strands 632:638 the N-terminal β strands 615:638 the N-terminal β strands of arrestin 615:650 Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. 28753425 5 49 theme phosphorylated 929:942 arg1 tail 967:970 the phosphorylated vasopressin-2 receptor tail 925:970 the phosphorylated vasopressin-2 receptor tail 925:970 These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. 28753425 6 50 theme phosphorylation 1052:1066 arg1 set 1045:1047 a set 1043:1047 a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs 1043:1169 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 6 50 theme phosphorylation 1052:1066 arg1 mechanism 1097:1105 a common mechanism 1088:1105 a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs 1088:1169 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 28753425 6 50 theme phosphorylation 1052:1066 arg1 phosphorylation 1052:1066 phosphorylation codes 1052:1072 phosphorylation codes 1052:1072 Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs. 21422170 5 0 theme NKp30-B7-H6-activating 686:707 arg1 complex 709:715 the NKp30-B7-H6-activating complex 682:715 the NKp30-B7-H6-activating complex 682:715 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 3 1 theme family 417:422 arg1 B7-H6 391:395 B7-H6 391:395 B7-H6 391:395 NKp30 recognizes B7-H6, a member of the B7 family which is expressed on tumor, but not healthy, cells. 21422170 3 1 theme family 417:422 arg1 member 400:405 a member 398:405 a member of the B7 family which is expressed on tumor, but not healthy, cells 398:474 NKp30 recognizes B7-H6, a member of the B7 family which is expressed on tumor, but not healthy, cells. 21422170 1 2 theme tumor 217:221 arg1 cells 223:227 tumor cells 217:227 tumor cells 217:227 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 5 3 theme inhibitory 787:796 arg1 complexes 798:806 the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes 752:806 complexes 798:806 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 8 4 theme NKp30-mediated 1314:1327 arg1 activity 1339:1346 NKp30-mediated cytolytic activity 1314:1346 NKp30-mediated cytolytic activity for tumor immunotherapy 1314:1370 This first structure of an NCR bound to ligand provides a template for designing molecules to stimulate NKp30-mediated cytolytic activity for tumor immunotherapy. 21422170 7 5 from contacts 1041:1048 arg1 interaction 1161:1171 an antibody-like interaction 1144:1171 an antibody-like interaction that is not observed for B7 or PD-L 1144:1207 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 5 6 theme T 780:780 arg1 complexes 798:806 the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes 752:806 complexes 798:806 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 1 7 theme cells 223:227 arg1 elimination 202:212 the elimination 198:212 the elimination of tumor cells 198:227 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 0 8 theme cell 89:92 arg1 B7-H6 101:105 its tumor cell ligand B7-H6 79:105 its tumor cell ligand B7-H6 79:105 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 2 9 theme activating 245:254 arg1 NCRs 288:291 NCRs 288:291 NCRs 288:291 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 9 theme activating 245:254 arg1 receptors 277:285 the activating natural cytotoxicity receptors 241:285 the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 241:316 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 3 10 theme healthy 461:467 arg1 tumor 446:450 tumor 446:450 tumor 446:450 NKp30 recognizes B7-H6, a member of the B7 family which is expressed on tumor, but not healthy, cells. 21422170 3 10 theme healthy 461:467 arg1 cells 470:474 but not healthy, cells 453:474 but not healthy, cells 453:474 NKp30 recognizes B7-H6, a member of the B7 family which is expressed on tumor, but not healthy, cells. 21422170 0 11 theme tumor 83:87 arg1 B7-H6 101:105 its tumor cell ligand B7-H6 79:105 its tumor cell ligand B7-H6 79:105 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 5 12 theme PD-1-PD-L 770:778 arg1 complexes 798:806 the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes 752:806 complexes 798:806 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 8 13 theme cytolytic 1329:1337 arg1 activity 1339:1346 NKp30-mediated cytolytic activity 1314:1346 NKp30-mediated cytolytic activity for tumor immunotherapy 1314:1370 This first structure of an NCR bound to ligand provides a template for designing molecules to stimulate NKp30-mediated cytolytic activity for tumor immunotherapy. 21422170 5 14 theme complex 709:715 arg1 organization 666:677 The overall organization 654:677 The overall organization of the NKp30-B7-H6-activating complex 654:715 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 1 15 theme system 171:176 arg1 lymphocytes 138:148 lymphocytes 138:148 lymphocytes of the innate immune system that participate in the elimination of tumor cells 138:227 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 1 15 theme system 171:176 arg1 cells 128:132 Natural killer (NK) cells 108:132 Natural killer (NK) cells 108:132 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 0 16 theme ligand 94:99 arg1 B7-H6 101:105 its tumor cell ligand B7-H6 79:105 its tumor cell ligand B7-H6 79:105 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 8 17 theme tumor 1352:1356 arg1 immunotherapy 1358:1370 tumor immunotherapy 1352:1370 tumor immunotherapy 1352:1370 This first structure of an NCR bound to ligand provides a template for designing molecules to stimulate NKp30-mediated cytolytic activity for tumor immunotherapy. 21422170 6 18 theme B7-H6 969:973 arg1 engagement 955:964 engagement 955:964 engagement of B7-H6 955:973 Whereas CTLA-4 and PD-1 use only the front β-sheet of their Ig-like domain to bind ligands, NKp30 uses both front and back β-sheets, resulting in engagement of B7-H6 via the side, as well as face, of the β-sandwich. 21422170 7 19 theme V-like 1127:1132 arg1 domain 1134:1139 its V-like domain 1123:1139 its V-like domain 1123:1139 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 2 20 theme major 325:329 arg1 role 331:334 a major role 323:334 a major role 323:334 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 5 21 theme overall 658:664 arg1 organization 666:677 The overall organization 654:677 The overall organization of the NKp30-B7-H6-activating complex 654:715 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 6 22 theme front 846:850 arg1 β-sheet 852:858 the front β-sheet 842:858 the front β-sheet of their Ig-like domain to bind ligands 842:898 Whereas CTLA-4 and PD-1 use only the front β-sheet of their Ig-like domain to bind ligands, NKp30 uses both front and back β-sheets, resulting in engagement of B7-H6 via the side, as well as face, of the β-sandwich. 21422170 7 23 theme domain 1134:1139 arg1 loops 1114:1118 the complementarity-determining region (CDR)-like loops 1064:1118 the complementarity-determining region (CDR)-like loops of its V-like domain 1064:1139 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 4 24 with complex 634:640 arg1 B7-H6 647:651 B7-H6 647:651 B7-H6 647:651 To understand the basis for tumor surveillance by NCRs, we determined the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1, in complex with B7-H6. 21422170 6 25 theme Ig-like 869:875 arg1 domain 877:882 their Ig-like domain to bind ligands 863:898 their Ig-like domain to bind ligands 863:898 Whereas CTLA-4 and PD-1 use only the front β-sheet of their Ig-like domain to bind ligands, NKp30 uses both front and back β-sheets, resulting in engagement of B7-H6 via the side, as well as face, of the β-sandwich. 21422170 1 26 theme innate 157:162 arg1 system 171:176 the innate immune system 153:176 the innate immune system 153:176 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 0 27 theme human 17:21 arg1 receptor 55:62 the human activating natural cytotoxicity receptor NKp30 13:68 the human activating natural cytotoxicity receptor NKp30 13:68 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 8 28 theme NCR 1237:1239 arg1 structure 1221:1229 This first structure 1210:1229 This first structure of an NCR bound to ligand 1210:1255 This first structure of an NCR bound to ligand provides a template for designing molecules to stimulate NKp30-mediated cytolytic activity for tumor immunotherapy. 21422170 2 29 theme cytotoxicity 264:275 arg1 NCRs 288:291 NCRs 288:291 NCRs 288:291 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 29 theme cytotoxicity 264:275 arg1 receptors 277:285 the activating natural cytotoxicity receptors 241:285 the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 241:316 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 30 theme tumor 356:360 arg1 lysis 367:371 NK cell-mediated tumor cell lysis 339:371 NK cell-mediated tumor cell lysis 339:371 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 31 theme cell 362:365 arg1 lysis 367:371 NK cell-mediated tumor cell lysis 339:371 NK cell-mediated tumor cell lysis 339:371 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 32 theme natural 256:262 arg1 NCRs 288:291 NCRs 288:291 NCRs 288:291 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 32 theme natural 256:262 arg1 receptors 277:285 the activating natural cytotoxicity receptors 241:285 the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 241:316 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 2 33 theme cell-mediated 342:354 arg1 lysis 367:371 NK cell-mediated tumor cell lysis 339:371 NK cell-mediated tumor cell lysis 339:371 In humans, the activating natural cytotoxicity receptors (NCRs) NKp30, NKp44, and NKp46 play a major role in NK cell-mediated tumor cell lysis. 21422170 0 34 theme natural 34:40 arg1 receptor 55:62 the human activating natural cytotoxicity receptor NKp30 13:68 the human activating natural cytotoxicity receptor NKp30 13:68 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 3 35 theme B7 414:415 arg1 family 417:422 the B7 family 410:422 the B7 family 410:422 NKp30 recognizes B7-H6, a member of the B7 family which is expressed on tumor, but not healthy, cells. 21422170 0 36 theme activating 23:32 arg1 receptor 55:62 the human activating natural cytotoxicity receptor NKp30 13:68 the human activating natural cytotoxicity receptor NKp30 13:68 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 0 37 theme receptor 55:62 arg1 Structure 0:8 Structure 0:8 Structure of the human activating natural cytotoxicity receptor NKp30 0:68 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 8 38 theme first 1215:1219 arg1 structure 1221:1229 This first structure 1210:1229 This first structure of an NCR bound to ligand 1210:1255 This first structure of an NCR bound to ligand provides a template for designing molecules to stimulate NKp30-mediated cytolytic activity for tumor immunotherapy. 21422170 5 39 theme cell 782:785 arg1 complexes 798:806 the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes 752:806 complexes 798:806 The overall organization of the NKp30-B7-H6-activating complex differs considerably from those of the CTLA-4-B7 and PD-1-PD-L T cell inhibitory complexes. 21422170 0 40 theme cytotoxicity 42:53 arg1 receptor 55:62 the human activating natural cytotoxicity receptor NKp30 13:68 the human activating natural cytotoxicity receptor NKp30 13:68 Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6. 21422170 1 41 theme Natural 108:114 arg1 killer 116:121 Natural killer 108:121 Natural killer (NK) cells 108:132 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 1 41 theme Natural 108:114 arg1 NK 124:125 NK 124:125 NK 124:125 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 1 42 theme killer 116:121 arg1 lymphocytes 138:148 lymphocytes 138:148 lymphocytes of the innate immune system that participate in the elimination of tumor cells 138:227 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 1 42 theme killer 116:121 arg1 cells 128:132 Natural killer (NK) cells 108:132 Natural killer (NK) cells 108:132 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 7 43 theme -like 1108:1112 arg1 loops 1114:1118 the complementarity-determining region (CDR)-like loops 1064:1118 the complementarity-determining region (CDR)-like loops of its V-like domain 1064:1139 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 4 44 theme CD28 587:590 arg1 family 592:597 the CD28 family 583:597 the CD28 family 583:597 To understand the basis for tumor surveillance by NCRs, we determined the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1, in complex with B7-H6. 21422170 4 45 theme family 592:597 arg1 NKp30 564:568 NKp30 564:568 NKp30 564:568 To understand the basis for tumor surveillance by NCRs, we determined the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1, in complex with B7-H6. 21422170 4 45 theme family 592:597 arg1 member 573:578 a member 571:578 a member of the CD28 family which includes CTLA-4 and PD-1 571:628 To understand the basis for tumor surveillance by NCRs, we determined the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1, in complex with B7-H6. 21422170 6 46 theme β-sandwich 1013:1022 arg1 face 1000:1003 face 1000:1003 face 1000:1003 Whereas CTLA-4 and PD-1 use only the front β-sheet of their Ig-like domain to bind ligands, NKp30 uses both front and back β-sheets, resulting in engagement of B7-H6 via the side, as well as face, of the β-sandwich. 21422170 6 46 theme β-sandwich 1013:1022 arg1 side 983:986 side 983:986 side 983:986 Whereas CTLA-4 and PD-1 use only the front β-sheet of their Ig-like domain to bind ligands, NKp30 uses both front and back β-sheets, resulting in engagement of B7-H6 via the side, as well as face, of the β-sandwich. 21422170 1 47 theme immune 164:169 arg1 system 171:176 the innate immune system 153:176 the innate immune system 153:176 Natural killer (NK) cells are lymphocytes of the innate immune system that participate in the elimination of tumor cells. 21422170 7 48 theme complementarity-determining 1068:1094 arg1 CDR 1104:1106 CDR 1104:1106 CDR 1104:1106 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 7 48 theme complementarity-determining 1068:1094 arg1 region 1096:1101 the complementarity-determining region 1064:1101 the complementarity-determining region (CDR) 1064:1107 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 7 49 theme antibody-like 1147:1159 arg1 interaction 1161:1171 an antibody-like interaction 1144:1171 an antibody-like interaction that is not observed for B7 or PD-L 1144:1207 Moreover, B7-H6 contacts NKp30 through the complementarity-determining region (CDR)-like loops of its V-like domain in an antibody-like interaction that is not observed for B7 or PD-L. 21422170 6 50 theme domain 877:882 arg1 β-sheet 852:858 the front β-sheet 842:858 the front β-sheet of their Ig-like domain to bind ligands 842:898 Whereas CTLA-4 and PD-1 use only the front β-sheet of their Ig-like domain to bind ligands, NKp30 uses both front and back β-sheets, resulting in engagement of B7-H6 via the side, as well as face, of the β-sandwich. 21422170 4 51 theme NKp30 564:568 arg1 structure 551:559 the structure 547:559 the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1 547:628 To understand the basis for tumor surveillance by NCRs, we determined the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1, in complex with B7-H6. 21422170 4 52 theme tumor 505:509 arg1 surveillance 511:522 tumor surveillance 505:522 tumor surveillance by NCRs 505:530 To understand the basis for tumor surveillance by NCRs, we determined the structure of NKp30, a member of the CD28 family which includes CTLA-4 and PD-1, in complex with B7-H6. 35835865 3 0 theme Rhesus 478:483 arg1 ankyrin-1 463:471 ankyrin-1 463:471 ankyrin-1 463:471 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 0 theme Rhesus 478:483 arg1 proteins 485:492 the Rhesus proteins RhAG and RhCE 474:506 the Rhesus proteins RhAG and RhCE 474:506 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 0 theme Rhesus 478:483 arg1 RhCE 503:506 RhCE 503:506 RhCE 503:506 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 0 theme Rhesus 478:483 arg1 RhAG 494:497 RhAG 494:497 RhAG 494:497 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 4 1 theme water 701:705 arg1 aquaporin-1 715:725 the water channel aquaporin-1 697:725 the water channel aquaporin-1 697:725 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 1 2 gly glycoproteins 300:312 arg1 glycoproteins 300:312 the Rhesus glycoproteins 289:312 the Rhesus glycoproteins 289:312 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 1 3 theme association 241:251 arg1 nature 227:232 the nature 223:232 the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins 223:312 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 5 4 theme membrane 885:892 arg1 clustering 902:911 ankyrin-mediated membrane protein clustering 868:911 ankyrin-mediated membrane protein clustering 868:911 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 3 5 theme core 447:450 arg1 complex 452:458 a core complex 445:458 a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B 445:580 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 2 6 theme complexes 371:379 arg1 structures 347:356 structures 347:356 structures of ankyrin-1 complexes purified from human erythrocytes 347:412 Here we present structures of ankyrin-1 complexes purified from human erythrocytes. 35835865 5 7 theme ankyrin-mediated 868:883 arg1 clustering 902:911 ankyrin-mediated membrane protein clustering 868:911 ankyrin-mediated membrane protein clustering 868:911 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 1 8 dep provided 112:119 arg1 tethers 158:164 tethers 158:164 tethers the spectrin-actin cytoskeleton to the lipid bilayer 158:217 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 1 8 dep provided 112:119 arg1 remains 314:320 remains 314:320 remains unknown 314:328 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 2 9 theme ankyrin-1 361:369 arg1 complexes 371:379 ankyrin-1 complexes 361:379 ankyrin-1 complexes purified from human erythrocytes 361:412 Here we present structures of ankyrin-1 complexes purified from human erythrocytes. 35835865 5 10 theme molecular 762:770 arg1 details 772:778 the molecular details 758:778 the molecular details of ankyrin-1 association with the erythrocyte membrane 758:833 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 4 11 theme T-shaped 596:603 arg1 conformation 605:616 The distinct T-shaped conformation 583:616 The distinct T-shaped conformation of membrane-bound ankyrin-1 583:644 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 3 12 theme anion 520:524 arg1 ankyrin-1 463:471 ankyrin-1 463:471 ankyrin-1 463:471 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 12 theme anion 520:524 arg1 exchanger 526:534 the band 3 anion exchanger 509:534 the band 3 anion exchanger 509:534 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 4 13 theme membrane-bound 621:634 arg1 ankyrin-1 636:644 membrane-bound ankyrin-1 621:644 membrane-bound ankyrin-1 621:644 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 0 14 theme human 20:24 arg1 complex 48:54 the human erythrocyte ankyrin-1 complex 16:54 the human erythrocyte ankyrin-1 complex 16:54 Architecture of the human erythrocyte ankyrin-1 complex. 35835865 1 15 theme erythrocyte 88:98 arg1 membrane 100:107 the erythrocyte membrane 84:107 the erythrocyte membrane 84:107 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 1 16 theme spectrin-actin 170:183 arg1 cytoskeleton 185:196 the spectrin-actin cytoskeleton 166:196 the spectrin-actin cytoskeleton 166:196 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 5 17 theme ankyrin-1 783:791 arg1 association 793:803 ankyrin-1 association 783:803 ankyrin-1 association with the erythrocyte membrane 783:833 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 1 18 theme band 262:265 arg1 exchanger 275:283 the band 3 anion exchanger 258:283 the band 3 anion exchanger 258:283 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 1 19 theme membrane 100:107 arg1 shape 75:79 shape 75:79 shape 75:79 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 1 19 theme membrane 100:107 arg1 stability 61:69 stability 61:69 stability 61:69 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 2 20 theme human 395:399 arg1 erythrocytes 401:412 human erythrocytes 395:412 human erythrocytes 395:412 Here we present structures of ankyrin-1 complexes purified from human erythrocytes. 35835865 0 21 theme ankyrin-1 38:46 arg1 complex 48:54 the human erythrocyte ankyrin-1 complex 16:54 the human erythrocyte ankyrin-1 complex 16:54 Architecture of the human erythrocyte ankyrin-1 complex. 35835865 5 22 theme protein 894:900 arg1 clustering 902:911 ankyrin-mediated membrane protein clustering 868:911 ankyrin-mediated membrane protein clustering 868:911 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 5 23 with association 793:803 arg1 membrane 826:833 the erythrocyte membrane 810:833 the erythrocyte membrane 810:833 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 4 24 theme distinct 587:594 arg1 conformation 605:616 The distinct T-shaped conformation 583:616 The distinct T-shaped conformation of membrane-bound ankyrin-1 583:644 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 0 25 theme erythrocyte 26:36 arg1 complex 48:54 the human erythrocyte ankyrin-1 complex 16:54 the human erythrocyte ankyrin-1 complex 16:54 Architecture of the human erythrocyte ankyrin-1 complex. 35835865 1 26 theme anion 269:273 arg1 exchanger 275:283 the band 3 anion exchanger 258:283 the band 3 anion exchanger 258:283 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 4 27 theme channel 707:713 arg1 aquaporin-1 715:725 the water channel aquaporin-1 697:725 the water channel aquaporin-1 697:725 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 1 28 with association 241:251 arg1 exchanger 275:283 the band 3 anion exchanger 258:283 the band 3 anion exchanger 258:283 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 1 28 with association 241:251 arg1 glycoproteins 300:312 the Rhesus glycoproteins 289:312 the Rhesus glycoproteins 289:312 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 3 29 theme ankyrin-1 463:471 arg1 complex 452:458 a core complex 445:458 a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B 445:580 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 0 30 theme complex 48:54 arg1 Architecture 0:11 Architecture 0:11 Architecture of the human erythrocyte ankyrin-1 complex 0:54 Architecture of the human erythrocyte ankyrin-1 complex. 35835865 4 31 theme RhCE 673:676 arg1 recognition 658:668 recognition 658:668 recognition of RhCE 658:676 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 4 31 theme RhCE 673:676 arg1 aquaporin-1 715:725 the water channel aquaporin-1 697:725 the water channel aquaporin-1 697:725 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 35835865 3 32 theme complex 452:458 arg1 architecture 429:440 the architecture 425:440 the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B 425:580 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 33 theme band 513:516 arg1 ankyrin-1 463:471 ankyrin-1 463:471 ankyrin-1 463:471 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 33 theme band 513:516 arg1 exchanger 526:534 the band 3 anion exchanger 509:534 the band 3 anion exchanger 509:534 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 1 34 theme lipid 205:209 arg1 bilayer 211:217 the lipid bilayer 201:217 the lipid bilayer 201:217 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 3 35 theme glycophorin 568:578 arg1 B 580:580 glycophorin B 568:580 glycophorin B 568:580 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 1 36 dep stability 61:69 arg1 The 57:59 The 57:59 The 57:59 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 5 37 theme clustering 902:911 arg1 mechanism 855:863 the mechanism 851:863 the mechanism of ankyrin-mediated membrane protein clustering 851:911 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 5 38 theme erythrocyte 814:824 arg1 membrane 826:833 the erythrocyte membrane 810:833 the erythrocyte membrane 810:833 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 3 39 theme glycophorin 550:560 arg1 A 562:562 glycophorin A 550:562 glycophorin A 550:562 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 5 40 theme association 793:803 arg1 details 772:778 the molecular details 758:778 the molecular details of ankyrin-1 association with the erythrocyte membrane 758:833 Together, our results uncover the molecular details of ankyrin-1 association with the erythrocyte membrane, and illustrate the mechanism of ankyrin-mediated membrane protein clustering. 35835865 1 41 theme ankyrin-1 128:136 arg1 complex 138:144 the ankyrin-1 complex 124:144 the ankyrin-1 complex 124:144 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 3 42 dep protein 537:543 arg1 A 562:562 glycophorin A 550:562 glycophorin A 550:562 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 42 dep protein 537:543 arg1 B 580:580 glycophorin B 568:580 glycophorin B 568:580 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 1 43 theme Rhesus 293:298 arg1 glycoproteins 300:312 the Rhesus glycoproteins 289:312 the Rhesus glycoproteins 289:312 The stability and shape of the erythrocyte membrane is provided by the ankyrin-1 complex, but how it tethers the spectrin-actin cytoskeleton to the lipid bilayer and the nature of its association with the band 3 anion exchanger and the Rhesus glycoproteins remains unknown. 35835865 3 44 dep proteins 485:492 arg1 proteins 485:492 the Rhesus proteins RhAG and RhCE 474:506 the Rhesus proteins RhAG and RhCE 474:506 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 44 dep proteins 485:492 arg1 RhCE 503:506 RhCE 503:506 RhCE 503:506 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 3 44 dep proteins 485:492 arg1 RhAG 494:497 RhAG 494:497 RhAG 494:497 We reveal the architecture of a core complex of ankyrin-1, the Rhesus proteins RhAG and RhCE, the band 3 anion exchanger, protein 4.2, glycophorin A and glycophorin B. 35835865 4 45 theme ankyrin-1 636:644 arg1 conformation 605:616 The distinct T-shaped conformation 583:616 The distinct T-shaped conformation of membrane-bound ankyrin-1 583:644 The distinct T-shaped conformation of membrane-bound ankyrin-1 facilitates recognition of RhCE and, unexpectedly, the water channel aquaporin-1. 24121512 3 0 theme cytosolic 599:607 arg1 NBCe1 619:623 NBCe1 619:623 NBCe1 (SLC4A4) 619:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 3 0 theme cytosolic 599:607 arg1 domain 609:614 the cytosolic domain 595:614 the cytosolic domain of NBCe1 (SLC4A4) 595:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 0 1 theme transporters 131:142 arg1 family 109:114 the solute carrier 4 (SLC4) family 81:114 the solute carrier 4 (SLC4) family of bicarbonate transporters 81:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 4 2 from arginine 860:867 arg1 mutations 832:840 single and double mutations 814:840 single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) 814:920 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 2 from arginine 860:867 arg1 Introduction 798:809 Introduction 798:809 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) 798:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 1 3 theme founding 192:199 arg1 member 201:206 the founding member 188:206 the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs) 188:356 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 1 3 theme founding 192:199 arg1 exchanger 151:159 Anion exchanger 1 145:161 Anion exchanger 1 (AE1; Band 3; SLC4A1) 145:183 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 9 4 theme cytosolic 1688:1696 arg1 domain 1698:1703 the cytosolic domain 1684:1703 the cytosolic domain 1684:1703 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 6 5 theme extensive 1346:1354 arg1 network 1372:1378 an extensive hydrogen-bonded network 1343:1378 an extensive hydrogen-bonded network involving Arg(283) and Glu(85) 1343:1409 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 3 6 theme proximal 647:654 arg1 acidosis 670:677 proximal renal tubular acidosis 647:677 proximal renal tubular acidosis 647:677 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 2 7 theme carboxyl-terminal 468:484 arg1 domain 495:500 a carboxyl-terminal membrane domain 466:500 a carboxyl-terminal membrane domain that carries out the transport function 466:540 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 0 8 theme carrier 92:98 arg1 family 109:114 the solute carrier 4 (SLC4) family 81:114 the solute carrier 4 (SLC4) family of bicarbonate transporters 81:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 4 9 theme AE1 955:957 arg1 AE1 955:957 human AE1 949:957 human AE1 (cdAE1) 949:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 9 theme AE1 955:957 arg1 domain 939:944 the cytosolic domain 925:944 the cytosolic domain of human AE1 (cdAE1) 925:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 1 10 theme Na 317:318 arg1 NBCs 352:355 NBCs 352:355 NBCs 352:355 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 1 10 theme Na 317:318 arg1 co-transporters 335:349 Na(+)-bicarbonate co-transporters 317:349 Na(+)-bicarbonate co-transporters (NBCs) 317:356 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 4 11 theme surface 993:999 arg1 expression 1001:1010 the cell surface expression 984:1010 the cell surface expression 984:1010 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 3 12 theme tubular 662:668 arg1 acidosis 670:677 proximal renal tubular acidosis 647:677 proximal renal tubular acidosis 647:677 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 8 13 theme AE1 1588:1590 arg1 expression 1574:1583 functional expression 1563:1583 functional expression of AE1 1563:1590 As these structural alterations did not impair functional expression of AE1, the cytosolic and membrane domains operate independently. 24121512 4 14 theme single 814:819 arg1 mutations 832:840 single and double mutations 814:840 single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) 814:920 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 9 15 theme SLC4 1776:1779 arg1 family 1781:1786 the SLC4 family 1772:1786 the SLC4 family of bicarbonate transporters 1772:1814 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 5 16 theme membrane 1090:1097 arg1 AE1 1109:1111 AE1 1109:1111 AE1 (mdAE1) 1109:1119 In addition, the membrane domain of AE1 (mdAE1) efficiently mediated anion transport. 24121512 5 16 theme membrane 1090:1097 arg1 domain 1099:1104 the membrane domain 1086:1104 the membrane domain of AE1 (mdAE1) 1086:1119 In addition, the membrane domain of AE1 (mdAE1) efficiently mediated anion transport. 24121512 3 17 theme transport 703:711 arg1 function 713:720 impaired transport function 694:720 impaired transport function 694:720 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 1 18 theme -bicarbonate 322:333 arg1 NBCs 352:355 NBCs 352:355 NBCs 352:355 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 1 18 theme -bicarbonate 322:333 arg1 co-transporters 335:349 Na(+)-bicarbonate co-transporters 317:349 Na(+)-bicarbonate co-transporters (NBCs) 317:356 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 3 19 theme cytosolic 743:751 arg1 domain 753:758 the cytosolic domain 739:758 the cytosolic domain 739:758 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 2 20 theme cytosolic 412:420 arg1 domain 422:427 an amino-terminal cytosolic domain 394:427 an amino-terminal cytosolic domain involved in protein interactions 394:460 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 6 21 theme cdAE1 1228:1232 arg1 55-356 1218:1223 55-356 1218:1223 55-356 1218:1223 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 6 21 theme cdAE1 1228:1232 arg1 cdAE1 1228:1232 cdAE1 1228:1232 cdAE1 1228:1232 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 3 22 theme substrate 776:784 arg1 permeation 786:795 substrate permeation 776:795 substrate permeation 776:795 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 9 23 theme transporters 1803:1814 arg1 family 1781:1786 the SLC4 family 1772:1786 the SLC4 family of bicarbonate transporters 1772:1814 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 6 24 theme crystal 1178:1184 arg1 structure 1186:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 4 25 from Introduction 798:809 arg1 Arg 870:872 Arg(283) 870:877 Arg(283) 870:877 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 25 from Introduction 798:809 arg1 AE1 955:957 human AE1 949:957 human AE1 (cdAE1) 949:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 25 from Introduction 798:809 arg1 glutamate 902:910 an interacting glutamate 887:910 an interacting glutamate (Glu(85)) 887:920 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 25 from Introduction 798:809 arg1 Glu 913:915 Glu(85) 913:919 Glu(85) 913:919 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 25 from Introduction 798:809 arg1 arginine 860:867 the equivalent arginine 845:867 the equivalent arginine (Arg(283)) 845:878 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 25 from Introduction 798:809 arg1 domain 939:944 the cytosolic domain 925:944 the cytosolic domain of human AE1 (cdAE1) 925:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 2 26 theme protein 441:447 arg1 interactions 449:460 protein interactions 441:460 protein interactions 441:460 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 6 27 theme 2.1-Å 1161:1165 arg1 structure 1186:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 1 28 theme transporters 261:272 arg1 family 239:244 the solute carrier 4 (SLC4) family 211:244 the solute carrier 4 (SLC4) family of bicarbonate transporters 211:272 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 9 29 from present 1712:1718 arg1 AE1 1723:1725 AE1 1723:1725 AE1 1723:1725 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 4 30 theme HEK-293 1058:1064 arg1 cells 1066:1070 HEK-293 cells 1058:1070 HEK-293 cells 1058:1070 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 3 31 from Mutation 543:550 arg1 NBCe1 619:623 NBCe1 619:623 NBCe1 (SLC4A4) 619:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 3 31 from Mutation 543:550 arg1 domain 609:614 the cytosolic domain 595:614 the cytosolic domain of NBCe1 (SLC4A4) 595:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 4 32 theme transport 1019:1027 arg1 activity 1029:1036 the transport activity 1015:1036 the transport activity of AE1 expressed in HEK-293 cells 1015:1070 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 7 33 theme conformational 1466:1479 arg1 changes 1481:1487 the pH-dependent conformational changes 1449:1487 the pH-dependent conformational changes 1449:1487 Mutations at these residues affected the pH-dependent conformational changes and stability of cdΔ54AE1. 24121512 0 34 theme access 12:17 arg1 tunnel 19:24 A substrate access tunnel 0:24 A substrate access tunnel in the cytosolic domain 0:48 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 0 34 theme access 12:17 arg1 feature 70:76 an essential feature 57:76 an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters 57:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 4 35 from glutamate 902:910 arg1 mutations 832:840 single and double mutations 814:840 single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) 814:920 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 35 from glutamate 902:910 arg1 Introduction 798:809 Introduction 798:809 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) 798:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 1 36 theme carrier 222:228 arg1 family 239:244 the solute carrier 4 (SLC4) family 211:244 the solute carrier 4 (SLC4) family of bicarbonate transporters 211:272 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 2 37 theme membrane 365:372 arg1 proteins 374:381 These membrane proteins 359:381 These membrane proteins 359:381 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 3 38 theme arginine 567:574 arg1 residue 576:582 a conserved arginine residue 555:582 a conserved arginine residue (R298S) 555:590 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 3 38 theme arginine 567:574 arg1 R298S 585:589 R298S 585:589 R298S 585:589 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 0 39 theme cytosolic 33:41 arg1 domain 43:48 the cytosolic domain 29:48 the cytosolic domain 29:48 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 1 40 dep AE1 164:166 arg1 Band 169:172 Band 3 169:174 AE1; Band 3; SLC4A1 164:182 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 1 40 dep AE1 164:166 arg1 SLC4A1 177:182 SLC4A1 177:182 AE1; Band 3; SLC4A1 164:182 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 9 41 theme bicarbonate 1791:1801 arg1 transporters 1803:1814 bicarbonate transporters 1791:1814 bicarbonate transporters 1791:1814 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 3 42 attach linked 637:642 arg1 acidosis 670:677 proximal renal tubular acidosis 647:677 proximal renal tubular acidosis 647:677 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 3 42 attach linked 637:642 arg2 Mutation 543:550 Mutation 543:550 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) 543:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 8 43 theme structural 1525:1534 arg1 alterations 1536:1546 these structural alterations 1519:1546 these structural alterations 1519:1546 As these structural alterations did not impair functional expression of AE1, the cytosolic and membrane domains operate independently. 24121512 9 44 theme substrate 1653:1661 arg1 tunnel 1670:1675 A substrate access tunnel 1651:1675 A substrate access tunnel within the cytosolic domain 1651:1703 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 44 theme substrate 1653:1661 arg1 present 1712:1718 present 1712:1718 present 1712:1718 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 3 45 theme NBCe1 619:623 arg1 NBCe1 619:623 NBCe1 619:623 NBCe1 (SLC4A4) 619:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 3 45 theme NBCe1 619:623 arg1 domain 609:614 the cytosolic domain 595:614 the cytosolic domain of NBCe1 (SLC4A4) 595:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 2 46 theme transport 523:531 arg1 function 533:540 the transport function 519:540 the transport function 519:540 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 9 47 located present 1712:1718 arg2 present 1712:1718 present 1712:1718 present 1712:1718 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 47 located present 1712:1718 arg2 tunnel 1670:1675 A substrate access tunnel 1651:1675 A substrate access tunnel within the cytosolic domain 1651:1703 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 47 located present 1712:1718 arg1 AE1 1723:1725 AE1 1723:1725 AE1 1723:1725 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 6 48 theme hydrogen-bonded 1356:1370 arg1 network 1372:1378 an extensive hydrogen-bonded network 1343:1378 an extensive hydrogen-bonded network involving Arg(283) and Glu(85) 1343:1409 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 1 49 theme Anion 145:149 arg1 member 201:206 the founding member 188:206 the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs) 188:356 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 1 49 theme Anion 145:149 arg1 exchanger 151:159 Anion exchanger 1 145:161 Anion exchanger 1 (AE1; Band 3; SLC4A1) 145:183 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 4 50 theme human 949:953 arg1 AE1 955:957 human AE1 949:957 human AE1 (cdAE1) 949:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 50 theme human 949:953 arg1 cdAE1 960:964 cdAE1 960:964 cdAE1 960:964 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 3 51 theme renal 656:660 arg1 acidosis 670:677 proximal renal tubular acidosis 647:677 proximal renal tubular acidosis 647:677 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 0 52 theme SLC4 103:106 arg1 family 109:114 the solute carrier 4 (SLC4) family 81:114 the solute carrier 4 (SLC4) family of bicarbonate transporters 81:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 6 53 theme physiological 1316:1328 arg1 pH 1330:1331 physiological pH 1316:1331 physiological pH 1316:1331 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 4 54 theme mutations 832:840 arg1 Introduction 798:809 Introduction 798:809 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) 798:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 55 theme equivalent 849:858 arg1 arginine 860:867 the equivalent arginine 845:867 the equivalent arginine (Arg(283)) 845:878 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 55 theme equivalent 849:858 arg1 Arg 870:872 Arg(283) 870:877 Arg(283) 870:877 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 0 56 theme family 109:114 arg1 tunnel 19:24 A substrate access tunnel 0:24 A substrate access tunnel in the cytosolic domain 0:48 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 0 56 theme family 109:114 arg1 feature 70:76 an essential feature 57:76 an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters 57:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 8 57 theme functional 1563:1572 arg1 expression 1574:1583 functional expression 1563:1583 functional expression of AE1 1563:1590 As these structural alterations did not impair functional expression of AE1, the cytosolic and membrane domains operate independently. 24121512 4 58 theme double 825:830 arg1 mutations 832:840 single and double mutations 814:840 single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) 814:920 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 0 59 theme bicarbonate 119:129 arg1 transporters 131:142 bicarbonate transporters 119:142 bicarbonate transporters 119:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 2 60 theme membrane 486:493 arg1 domain 495:500 a carboxyl-terminal membrane domain 466:500 a carboxyl-terminal membrane domain that carries out the transport function 466:540 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 9 61 theme family 1781:1786 arg1 feature 1761:1767 an essential feature 1748:1767 an essential feature of the SLC4 family of bicarbonate transporters 1748:1814 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 6 62 theme cdΔ54AE1 1199:1206 arg1 structure 1186:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 2 63 theme amino-terminal 397:410 arg1 domain 422:427 an amino-terminal cytosolic domain 394:427 an amino-terminal cytosolic domain involved in protein interactions 394:460 These membrane proteins consist of an amino-terminal cytosolic domain involved in protein interactions and a carboxyl-terminal membrane domain that carries out the transport function. 24121512 0 64 from tunnel 19:24 arg1 domain 43:48 the cytosolic domain 29:48 the cytosolic domain 29:48 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 9 65 from AE1 1723:1725 arg1 tunnel 1670:1675 A substrate access tunnel 1651:1675 A substrate access tunnel within the cytosolic domain 1651:1703 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 65 from AE1 1723:1725 arg1 present 1712:1718 present 1712:1718 present 1712:1718 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 4 66 theme cell 988:991 arg1 expression 1001:1010 the cell surface expression 984:1010 the cell surface expression 984:1010 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 3 67 theme impaired 694:701 arg1 function 713:720 impaired transport function 694:720 impaired transport function 694:720 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 0 68 theme essential 60:68 arg1 tunnel 19:24 A substrate access tunnel 0:24 A substrate access tunnel in the cytosolic domain 0:48 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 0 68 theme essential 60:68 arg1 feature 70:76 an essential feature 57:76 an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters 57:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 1 69 theme family 239:244 arg1 member 201:206 the founding member 188:206 the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs) 188:356 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 1 69 theme family 239:244 arg1 exchanger 151:159 Anion exchanger 1 145:161 Anion exchanger 1 (AE1; Band 3; SLC4A1) 145:183 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 8 70 theme cytosolic 1597:1605 arg1 domains 1620:1626 the cytosolic and membrane domains 1593:1626 domains 1620:1626 As these structural alterations did not impair functional expression of AE1, the cytosolic and membrane domains operate independently. 24121512 6 71 theme resolution 1167:1176 arg1 structure 1186:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure 1159:1194 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 1 72 theme bicarbonate 249:259 arg1 transporters 261:272 bicarbonate transporters 249:272 bicarbonate transporters 249:272 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 8 73 theme membrane 1611:1618 arg1 domains 1620:1626 the cytosolic and membrane domains 1593:1626 domains 1620:1626 As these structural alterations did not impair functional expression of AE1, the cytosolic and membrane domains operate independently. 24121512 0 74 theme solute 85:90 arg1 family 109:114 the solute carrier 4 (SLC4) family 81:114 the solute carrier 4 (SLC4) family of bicarbonate transporters 81:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 7 75 theme cdΔ54AE1 1506:1513 arg1 stability 1493:1501 stability 1493:1501 stability of cdΔ54AE1 1493:1513 Mutations at these residues affected the pH-dependent conformational changes and stability of cdΔ54AE1. 24121512 7 75 theme cdΔ54AE1 1506:1513 arg1 changes 1481:1487 the pH-dependent conformational changes 1449:1487 the pH-dependent conformational changes 1449:1487 Mutations at these residues affected the pH-dependent conformational changes and stability of cdΔ54AE1. 24121512 5 76 theme anion 1142:1146 arg1 transport 1148:1156 anion transport 1142:1156 anion transport 1142:1156 In addition, the membrane domain of AE1 (mdAE1) efficiently mediated anion transport. 24121512 0 77 theme substrate 2:10 arg1 tunnel 19:24 A substrate access tunnel 0:24 A substrate access tunnel in the cytosolic domain 0:48 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 0 77 theme substrate 2:10 arg1 feature 70:76 an essential feature 57:76 an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters 57:142 A substrate access tunnel in the cytosolic domain is not an essential feature of the solute carrier 4 (SLC4) family of bicarbonate transporters. 24121512 1 78 theme chloride/bicarbonate 288:307 arg1 AEs 309:311 chloride/bicarbonate AEs 288:311 chloride/bicarbonate AEs 288:311 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 7 79 from residues 1431:1438 arg1 Mutations 1412:1420 Mutations 1412:1420 Mutations at these residues 1412:1438 Mutations at these residues affected the pH-dependent conformational changes and stability of cdΔ54AE1. 24121512 4 80 theme interacting 890:900 arg1 glutamate 902:910 an interacting glutamate 887:910 an interacting glutamate (Glu(85)) 887:920 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 80 theme interacting 890:900 arg1 Glu 913:915 Glu(85) 913:919 Glu(85) 913:919 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 81 theme AE1 1041:1043 arg1 expression 1001:1010 the cell surface expression 984:1010 the cell surface expression 984:1010 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 81 theme AE1 1041:1043 arg1 activity 1029:1036 the transport activity 1015:1036 the transport activity of AE1 expressed in HEK-293 cells 1015:1070 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 5 82 theme AE1 1109:1111 arg1 AE1 1109:1111 AE1 1109:1111 AE1 (mdAE1) 1109:1119 In addition, the membrane domain of AE1 (mdAE1) efficiently mediated anion transport. 24121512 5 82 theme AE1 1109:1111 arg1 domain 1099:1104 the membrane domain 1086:1104 the membrane domain of AE1 (mdAE1) 1086:1119 In addition, the membrane domain of AE1 (mdAE1) efficiently mediated anion transport. 24121512 6 83 dep residues 1209:1216 arg1 55-356 1218:1223 55-356 1218:1223 55-356 1218:1223 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 6 83 dep residues 1209:1216 arg1 cdAE1 1228:1232 cdAE1 1228:1232 cdAE1 1228:1232 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 3 84 theme residue 576:582 arg1 Mutation 543:550 Mutation 543:550 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) 543:632 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 9 85 theme essential 1751:1759 arg1 feature 1761:1767 an essential feature 1748:1767 an essential feature of the SLC4 family of bicarbonate transporters 1748:1814 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 1 86 theme solute 215:220 arg1 family 239:244 the solute carrier 4 (SLC4) family 211:244 the solute carrier 4 (SLC4) family of bicarbonate transporters 211:272 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 7 87 theme pH-dependent 1453:1464 arg1 changes 1481:1487 the pH-dependent conformational changes 1449:1487 the pH-dependent conformational changes 1449:1487 Mutations at these residues affected the pH-dependent conformational changes and stability of cdΔ54AE1. 24121512 3 88 theme conserved 557:565 arg1 residue 576:582 a conserved arginine residue 555:582 a conserved arginine residue (R298S) 555:590 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 3 88 theme conserved 557:565 arg1 R298S 585:589 R298S 585:589 R298S 585:589 Mutation of a conserved arginine residue (R298S) in the cytosolic domain of NBCe1 (SLC4A4) is linked to proximal renal tubular acidosis and results in impaired transport function, suggesting that the cytosolic domain plays a role in substrate permeation. 24121512 1 89 dep exchanger 151:159 arg1 AE1 164:166 AE1 164:166 AE1; Band 3; SLC4A1 164:182 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 4 90 contain had 967:969 arg2 effect 974:979 no effect 971:979 no effect 971:979 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 90 contain had 967:969 arg1 Introduction 798:809 Introduction 798:809 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) 798:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 6 91 dep amino-terminal 1247:1260 arg1 regions 1295:1301 disordered regions 1284:1301 disordered regions 1284:1301 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 24121512 9 92 theme access 1663:1668 arg1 tunnel 1670:1675 A substrate access tunnel 1651:1675 A substrate access tunnel within the cytosolic domain 1651:1703 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 92 theme access 1663:1668 arg1 present 1712:1718 present 1712:1718 present 1712:1718 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 1 93 theme SLC4 233:236 arg1 family 239:244 the solute carrier 4 (SLC4) family 211:244 the solute carrier 4 (SLC4) family of bicarbonate transporters 211:272 Anion exchanger 1 (AE1; Band 3; SLC4A1) is the founding member of the solute carrier 4 (SLC4) family of bicarbonate transporters that includes chloride/bicarbonate AEs and Na(+)-bicarbonate co-transporters (NBCs). 24121512 9 94 attach present 1712:1718 arg2 present 1712:1718 present 1712:1718 present 1712:1718 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 94 attach present 1712:1718 arg2 tunnel 1670:1675 A substrate access tunnel 1651:1675 A substrate access tunnel within the cytosolic domain 1651:1703 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 9 94 attach present 1712:1718 arg1 AE1 1723:1725 AE1 1723:1725 AE1 1723:1725 A substrate access tunnel within the cytosolic domain is not present in AE1 and therefore is not an essential feature of the SLC4 family of bicarbonate transporters. 24121512 4 95 theme cytosolic 929:937 arg1 AE1 955:957 human AE1 949:957 human AE1 (cdAE1) 949:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 4 95 theme cytosolic 929:937 arg1 domain 939:944 the cytosolic domain 925:944 the cytosolic domain of human AE1 (cdAE1) 925:965 Introduction of single and double mutations at the equivalent arginine (Arg(283)) and at an interacting glutamate (Glu(85)) in the cytosolic domain of human AE1 (cdAE1) had no effect on the cell surface expression or the transport activity of AE1 expressed in HEK-293 cells. 24121512 6 96 theme disordered 1284:1293 arg1 regions 1295:1301 disordered regions 1284:1301 disordered regions 1284:1301 A 2.1-Å resolution crystal structure of cdΔ54AE1 (residues 55-356 of cdAE1) lacking the amino-terminal and carboxyl-terminal disordered regions, produced at physiological pH, revealed an extensive hydrogen-bonded network involving Arg(283) and Glu(85). 9857012 0 0 theme metabolism 89:98 arg1 glycogenin-2 26:37 human glycogenin-2 20:37 human glycogenin-2 20:37 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 0 0 theme metabolism 89:98 arg1 initiator 61:69 a self-glucosylating initiator 40:69 a self-glucosylating initiator of liver glycogen metabolism 40:98 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 11 1 from pellet 1255:1260 arg1 present 1230:1236 present 1230:1236 present 1230:1236 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 11 2 attach present 1230:1236 arg2 level 1212:1216 the level 1208:1216 the level of glycogen present in the low speed pellet 1208:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 11 2 attach present 1230:1236 arg1 pellet 1255:1260 the low speed pellet 1241:1260 the low speed pellet 1241:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 6 3 theme predominant 565:575 arg1 isoform 577:583 the predominant isoform 561:583 the predominant isoform in liver 561:592 Based on analysis by polymerase chain reaction, the predominant isoform in liver was glycogenin-2beta. 9857012 6 3 theme predominant 565:575 arg1 glycogenin-2beta 598:613 glycogenin-2beta 598:613 glycogenin-2beta 598:613 Based on analysis by polymerase chain reaction, the predominant isoform in liver was glycogenin-2beta. 9857012 0 4 theme glycogen 80:87 arg1 metabolism 89:98 liver glycogen metabolism 74:98 liver glycogen metabolism 74:98 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 12 5 theme glycogen 1399:1406 arg1 accumulation 1408:1419 glycogen accumulation 1399:1419 glycogen accumulation 1399:1419 This result is important since it indicates that the level of glycogenin-2 can determine glycogen accumulation and hence has the potential to control glycogen synthesis. 9857012 11 6 theme glycogen 1221:1228 arg1 level 1212:1216 the level 1208:1216 the level of glycogen present in the low speed pellet 1208:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 11 7 theme glycogenin-2alpha 1131:1147 arg1 overexpression 1113:1126 Stable overexpression 1106:1126 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells 1106:1173 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 11 8 theme fibroblast 1158:1167 arg1 cells 1169:1173 Rat-1 fibroblast cells 1152:1173 Rat-1 fibroblast cells 1152:1173 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 7 9 theme RD-ES 658:662 arg1 cells 664:668 Ewing's sarcoma RD-ES cells 642:668 Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate 642:732 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 9 10 theme in 877:878 arg1 interactions 886:897 in vitro interactions 877:897 in vitro interactions 877:897 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 11 11 theme present 1230:1236 arg1 level 1212:1216 the level 1208:1216 the level of glycogen present in the low speed pellet 1208:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 5 12 theme purifed 428:434 arg1 glycogen 444:451 purifed high Mr glycogen 428:451 purifed high Mr glycogen 428:451 Similarly, purifed high Mr glycogen was only detected after release by alpha-amylase treatment. 9857012 11 13 from increase 1196:1203 arg1 level 1212:1216 the level 1208:1216 the level of glycogen present in the low speed pellet 1208:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 9 14 from liver 931:935 arg1 co-immunoprecipitation 903:924 co-immunoprecipitation 903:924 co-immunoprecipitation from liver and cell extracts 903:953 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 9 14 from liver 931:935 arg1 interactions 886:897 in vitro interactions 877:897 in vitro interactions 877:897 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 8 15 theme RD-ES 762:766 arg1 extracts 773:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 5 16 theme high 436:439 arg1 glycogen 444:451 purifed high Mr glycogen 428:451 purifed high Mr glycogen 428:451 Similarly, purifed high Mr glycogen was only detected after release by alpha-amylase treatment. 9857012 7 17 located found 633:637 arg1 cells 664:668 Ewing's sarcoma RD-ES cells 642:668 Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate 642:732 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 7 17 located found 633:637 arg2 Glycogenin-2 616:627 Glycogenin-2 616:627 Glycogenin-2 616:627 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 11 18 theme speed 1291:1295 arg1 supernatant 1297:1307 the low speed supernatant 1283:1307 the low speed supernatant 1283:1307 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 8 19 contain contained 787:795 arg1 extracts 773:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 8 19 contain contained 787:795 arg2 glycogenin-1 797:808 glycogenin-1 797:808 glycogenin-1 797:808 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 4 20 from extracts 326:333 arg1 detectable 370:379 detectable 370:379 detectable 370:379 In human liver extracts, most of the glycogenin-2 was only detectable after treatment with alpha-amylase. 9857012 5 21 theme Mr 441:442 arg1 glycogen 444:451 purifed high Mr glycogen 428:451 purifed high Mr glycogen 428:451 Similarly, purifed high Mr glycogen was only detected after release by alpha-amylase treatment. 9857012 4 22 with treatment 387:395 arg1 alpha-amylase 402:414 alpha-amylase 402:414 alpha-amylase 402:414 In human liver extracts, most of the glycogenin-2 was only detectable after treatment with alpha-amylase. 9857012 11 23 theme low 1245:1247 arg1 pellet 1255:1260 the low speed pellet 1241:1260 the low speed pellet 1241:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 12 24 contain has 1431:1433 arg1 level 1363:1367 the level 1359:1367 the level of glycogenin-2 1359:1383 This result is important since it indicates that the level of glycogenin-2 can determine glycogen accumulation and hence has the potential to control glycogen synthesis. 9857012 12 24 contain has 1431:1433 arg2 potential 1439:1447 the potential to control glycogen synthesis 1435:1477 the potential to control glycogen synthesis 1435:1477 This result is important since it indicates that the level of glycogenin-2 can determine glycogen accumulation and hence has the potential to control glycogen synthesis. 9857012 10 25 theme Tyr-196 968:974 arg1 Mutation 956:963 Mutation 956:963 Mutation of Tyr-196 in glycogenin-2 to a Phe residue 956:1007 Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. 9857012 11 26 from change 1273:1278 arg1 supernatant 1297:1307 the low speed supernatant 1283:1307 the low speed supernatant 1283:1307 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 1 27 theme glycogen 207:214 arg1 biosynthesis 216:227 glycogen biosynthesis 207:227 glycogen biosynthesis 207:227 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 6 28 theme chain 545:549 arg1 reaction 551:558 polymerase chain reaction 534:558 polymerase chain reaction 534:558 Based on analysis by polymerase chain reaction, the predominant isoform in liver was glycogenin-2beta. 9857012 11 29 theme speed 1249:1253 arg1 pellet 1255:1260 the low speed pellet 1241:1260 the low speed pellet 1241:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 4 30 theme liver 320:324 arg1 extracts 326:333 human liver extracts 314:333 human liver extracts 314:333 In human liver extracts, most of the glycogenin-2 was only detectable after treatment with alpha-amylase. 9857012 1 31 theme biosynthesis 216:227 arg1 initiation 193:202 the initiation 189:202 the initiation of glycogen biosynthesis 189:227 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 0 32 theme glycogenin-2 26:37 arg1 Characterization 0:15 Characterization 0:15 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 0:99 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 11 33 theme 5-fold 1189:1194 arg1 increase 1196:1203 a 5-fold increase 1187:1203 a 5-fold increase in the level of glycogen present in the low speed pellet 1187:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 4 34 theme human 314:318 arg1 extracts 326:333 human liver extracts 314:333 human liver extracts 314:333 In human liver extracts, most of the glycogenin-2 was only detectable after treatment with alpha-amylase. 9857012 11 35 theme Rat-1 1152:1156 arg1 cells 1169:1173 Rat-1 fibroblast cells 1152:1173 Rat-1 fibroblast cells 1152:1173 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 0 36 theme human 20:24 arg1 glycogenin-2 26:37 human glycogenin-2 20:37 human glycogenin-2 20:37 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 0 36 theme human 20:24 arg1 initiator 61:69 a self-glucosylating initiator 40:69 a self-glucosylating initiator of liver glycogen metabolism 40:98 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 7 37 theme Mr 718:719 arg1 carbohydrate 721:732 high Mr carbohydrate 713:732 high Mr carbohydrate 713:732 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 1 38 dep involved 177:184 arg1 Mu 230:231 Mu 230:231 Mu 230:231 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 38 dep involved 177:184 arg1 protein 157:163 a recently described self-glucosylating protein 117:163 a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol 117:283 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 11 39 theme little 1266:1271 arg1 change 1273:1278 little change 1266:1278 little change in the low speed supernatant 1266:1307 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 9 40 dep one 855:857 arg1 another 859:865 another 859:865 another 859:865 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 7 41 theme sarcoma 650:656 arg1 cells 664:668 Ewing's sarcoma RD-ES cells 642:668 Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate 642:732 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 9 42 theme cell 941:944 arg1 extracts 946:953 cell extracts 941:953 cell extracts 941:953 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 4 43 from detectable 370:379 arg1 extracts 326:333 human liver extracts 314:333 human liver extracts 314:333 In human liver extracts, most of the glycogenin-2 was only detectable after treatment with alpha-amylase. 9857012 12 44 theme glycogen 1460:1467 arg1 synthesis 1469:1477 glycogen synthesis 1460:1477 glycogen synthesis 1460:1477 This result is important since it indicates that the level of glycogenin-2 can determine glycogen accumulation and hence has the potential to control glycogen synthesis. 9857012 6 45 theme polymerase 534:543 arg1 reaction 551:558 polymerase chain reaction 534:558 polymerase chain reaction 534:558 Based on analysis by polymerase chain reaction, the predominant isoform in liver was glycogenin-2beta. 9857012 12 46 theme glycogenin-2 1372:1383 arg1 level 1363:1367 the level 1359:1367 the level of glycogenin-2 1359:1383 This result is important since it indicates that the level of glycogenin-2 can determine glycogen accumulation and hence has the potential to control glycogen synthesis. 9857012 11 47 theme low 1287:1289 arg1 supernatant 1297:1307 the low speed supernatant 1283:1307 the low speed supernatant 1283:1307 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 7 48 theme high 713:716 arg1 carbohydrate 721:732 high Mr carbohydrate 713:732 high Mr carbohydrate 713:732 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 3 49 dep 272 292:294 arg1 27589-27597 297:307 27589-27597 297:307 27589-27597 297:307 272, 27589-27597). 9857012 1 50 dep Skurat 238:243 arg1 Biol 280:283 Biol 280:283 Biol 280:283 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 10 51 theme glycogenin-2 1034:1045 arg1 ability 1023:1029 the ability 1019:1029 the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1 1019:1103 Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. 9857012 0 52 theme self-glucosylating 42:59 arg1 glycogenin-2 26:37 human glycogenin-2 20:37 human glycogenin-2 20:37 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 0 52 theme self-glucosylating 42:59 arg1 initiator 61:69 a self-glucosylating initiator 40:69 a self-glucosylating initiator of liver glycogen metabolism 40:98 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 10 53 dep ability 1023:1029 arg1 interact 1078:1085 interact 1078:1085 not to interact with glycogenin-1 1071:1103 Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. 9857012 10 53 dep ability 1023:1029 arg1 self-glucosylate 1050:1065 self-glucosylate 1050:1065 to self-glucosylate 1047:1065 Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. 9857012 10 54 theme Phe 997:999 arg1 residue 1001:1007 a Phe residue 995:1007 a Phe residue 995:1007 Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. 9857012 5 55 theme alpha-amylase 488:500 arg1 treatment 502:510 alpha-amylase treatment 488:510 alpha-amylase treatment 488:510 Similarly, purifed high Mr glycogen was only detected after release by alpha-amylase treatment. 9857012 1 56 theme described 128:136 arg1 Mu 230:231 Mu 230:231 Mu 230:231 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 56 theme described 128:136 arg1 protein 157:163 a recently described self-glucosylating protein 117:163 a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol 117:283 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 56 theme described 128:136 arg1 Glycogenin-2 101:112 Glycogenin-2 101:112 Glycogenin-2 101:112 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 8 57 theme liver 746:750 arg1 extracts 773:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 7 58 dep cells 664:668 arg1 associated 697:706 associated 697:706 was not associated with high Mr carbohydrate 689:732 Glycogenin-2 was found in Ewing's sarcoma RD-ES cells where, however, it was not associated with high Mr carbohydrate. 9857012 1 59 theme self-glucosylating 138:155 arg1 Mu 230:231 Mu 230:231 Mu 230:231 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 59 theme self-glucosylating 138:155 arg1 protein 157:163 a recently described self-glucosylating protein 117:163 a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol 117:283 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 59 theme self-glucosylating 138:155 arg1 Glycogenin-2 101:112 Glycogenin-2 101:112 Glycogenin-2 101:112 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 8 60 theme human 740:744 arg1 liver 746:750 human liver 740:750 human liver 740:750 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 9 61 dep in 877:878 arg1 vitro 880:884 vitro 880:884 vitro 880:884 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 11 62 theme Stable 1106:1111 arg1 overexpression 1113:1126 Stable overexpression 1106:1126 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells 1106:1173 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 8 63 theme human 756:760 arg1 RD-ES 762:766 human RD-ES 756:766 human RD-ES 756:766 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 0 64 theme liver 74:78 arg1 metabolism 89:98 liver glycogen metabolism 74:98 liver glycogen metabolism 74:98 Characterization of human glycogenin-2, a self-glucosylating initiator of liver glycogen metabolism. 9857012 6 65 from isoform 577:583 arg1 liver 588:592 liver 588:592 liver 588:592 Based on analysis by polymerase chain reaction, the predominant isoform in liver was glycogenin-2beta. 9857012 11 66 from overexpression 1113:1126 arg1 cells 1169:1173 Rat-1 fibroblast cells 1152:1173 Rat-1 fibroblast cells 1152:1173 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 9857012 10 67 from Mutation 956:963 arg1 glycogenin-2 979:990 glycogenin-2 979:990 glycogenin-2 to a Phe residue 979:1007 Mutation of Tyr-196 in glycogenin-2 to a Phe residue abolished the ability of glycogenin-2 to self-glucosylate but not to interact with glycogenin-1. 9857012 9 68 from extracts 946:953 arg1 co-immunoprecipitation 903:924 co-immunoprecipitation 903:924 co-immunoprecipitation from liver and cell extracts 903:953 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 9 68 from extracts 946:953 arg1 interactions 886:897 in vitro interactions 877:897 in vitro interactions 877:897 Glycogenin-1 and glycogenin-2 interact with one another, based on in vitro interactions and co-immunoprecipitation from liver and cell extracts. 9857012 1 69 dep Mu 230:231 arg1 J 267:267 J 267:267 J 267:267 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 69 dep Mu 230:231 arg1 1997 271:274 1997 271:274 1997 271:274 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 69 dep Mu 230:231 arg1 Skurat 238:243 Skurat 238:243 Skurat 238:243 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 69 dep Mu 230:231 arg1 V. 249:250 V. 249:250 V. 249:250 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 1 69 dep Mu 230:231 arg1 P. 264:265 P. 264:265 P. 264:265 Glycogenin-2 is a recently described self-glucosylating protein potentially involved in the initiation of glycogen biosynthesis (Mu, J., Skurat, A. V., and Roach, P. J. (1997) J. Biol. 9857012 8 70 theme cell 768:771 arg1 extracts 773:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts 735:780 Both human liver and human RD-ES cell extracts also contained glycogenin-1. 9857012 11 71 from present 1230:1236 arg1 pellet 1255:1260 the low speed pellet 1241:1260 the low speed pellet 1241:1260 Stable overexpression of glycogenin-2alpha in Rat-1 fibroblast cells resulted in a 5-fold increase in the level of glycogen present in the low speed pellet but little change in the low speed supernatant. 29345911 7 0 theme HepG2 945:949 arg1 proliferation 956:968 HuH7 and HepG2 cell proliferation 936:968 HuH7 and HepG2 cell proliferation 936:968 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 2 1 theme furin-like 341:350 arg1 convertase 352:361 a furin-like convertase 339:361 a furin-like convertase 339:361 It is also N-glycosylated and processed by a furin-like convertase. 29345911 1 2 theme membrane 239:246 arg1 surface 217:223 the outer surface 207:223 the outer surface of the plasma membrane 207:246 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 6 3 theme glycosaminoglycan 860:876 arg1 chains 878:883 glycosaminoglycan chains 860:883 glycosaminoglycan chains 860:883 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 5 4 theme soluble 633:639 arg1 form 641:644 a soluble form 631:644 a soluble form of GPC3 after lentiviral transduction 631:682 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 9 5 theme cell 1141:1144 arg1 area 1154:1157 HuH7 cell nuclear area 1136:1157 HuH7 cell nuclear area 1136:1157 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 10 6 theme growth 1337:1342 arg1 inhibition 1344:1353 growth inhibition 1337:1353 growth inhibition 1337:1353 Unexpectedly, for both cell types, the levels of apoptosis, cell division, and β-catenin were not altered by sGPC3m, although growth inhibition was very efficient. 29345911 0 7 theme Hepatocellular 99:112 arg1 Cells 124:128 Hepatocellular Carcinoma Cells 99:128 Hepatocellular Carcinoma Cells 99:128 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 4 8 theme hepatocellular 503:516 arg1 HCC 529:531 HCC 529:531 HCC 529:531 Although GPC3 is undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). 29345911 4 8 theme hepatocellular 503:516 arg1 carcinoma 518:526 hepatocellular carcinoma 503:526 hepatocellular carcinoma (HCC) 503:532 Although GPC3 is undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). 29345911 6 9 theme mutant 785:790 arg1 sGPC3m 798:803 sGPC3m 798:803 sGPC3m 798:803 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 6 9 theme mutant 785:790 arg1 GPC3 792:795 a mutant GPC3 783:795 a mutant GPC3 783:795 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 10 10 theme division 1276:1283 arg1 levels 1250:1255 the levels 1246:1255 the levels of apoptosis, cell division, and β-catenin 1246:1298 Unexpectedly, for both cell types, the levels of apoptosis, cell division, and β-catenin were not altered by sGPC3m, although growth inhibition was very efficient. 29345911 10 11 theme β-catenin 1290:1298 arg1 levels 1250:1255 the levels 1246:1255 the levels of apoptosis, cell division, and β-catenin 1246:1298 Unexpectedly, for both cell types, the levels of apoptosis, cell division, and β-catenin were not altered by sGPC3m, although growth inhibition was very efficient. 29345911 6 12 theme more 695:698 arg1 insight 700:706 more insight 695:706 more insight into the role of some of its post-translational modifications 695:768 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 1 13 theme glycosylphosphatidylinositol 253:280 arg1 anchor 288:293 a glycosylphosphatidylinositol (GPI) anchor 251:293 a glycosylphosphatidylinositol (GPI) anchor 251:293 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 0 14 theme Cells 124:128 arg1 Proliferation 82:94 Proliferation 82:94 Proliferation of Hepatocellular Carcinoma Cells 82:128 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 10 15 theme cell 1234:1237 arg1 types 1239:1243 both cell types 1229:1243 both cell types 1229:1243 Unexpectedly, for both cell types, the levels of apoptosis, cell division, and β-catenin were not altered by sGPC3m, although growth inhibition was very efficient. 29345911 9 16 theme HuH7 1136:1139 arg1 area 1154:1157 HuH7 cell nuclear area 1136:1157 HuH7 cell nuclear area 1136:1157 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 1 17 theme complex 154:160 arg1 proteoglycan 178:189 a complex heparan sulfate proteoglycan 152:189 a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor 152:293 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 1 17 theme complex 154:160 arg1 Glypican 131:138 Glypican 3 131:140 Glypican 3 (GPC3) 131:147 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 0 18 theme Carcinoma 114:122 arg1 Cells 124:128 Hepatocellular Carcinoma Cells 99:128 Hepatocellular Carcinoma Cells 99:128 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 6 19 theme cleavage 841:848 arg1 site 850:853 convertase cleavage site 830:853 convertase cleavage site 830:853 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 6 19 theme cleavage 841:848 arg1 anchor 822:827 its GPI anchor 814:827 its GPI anchor 814:827 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 9 20 theme nuclear 1146:1152 arg1 area 1154:1157 HuH7 cell nuclear area 1136:1157 HuH7 cell nuclear area 1136:1157 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 3 21 contain has 369:371 arg1 GPC3 364:367 GPC3 364:367 GPC3 364:367 GPC3 has numerous biological functions. 29345911 3 21 contain has 369:371 arg2 functions 393:401 numerous biological functions 373:401 numerous biological functions 373:401 GPC3 has numerous biological functions. 29345911 1 22 theme heparan 162:168 arg1 proteoglycan 178:189 a complex heparan sulfate proteoglycan 152:189 a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor 152:293 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 1 22 theme heparan 162:168 arg1 Glypican 131:138 Glypican 3 131:140 Glypican 3 (GPC3) 131:147 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 3 23 theme numerous 373:380 arg1 functions 393:401 numerous biological functions 373:401 numerous biological functions 373:401 GPC3 has numerous biological functions. 29345911 6 24 theme convertase 830:839 arg1 site 850:853 convertase cleavage site 830:853 convertase cleavage site 830:853 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 6 24 theme convertase 830:839 arg1 anchor 822:827 its GPI anchor 814:827 its GPI anchor 814:827 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 5 25 theme cells 571:575 arg1 proliferation 550:562 proliferation 550:562 proliferation of HCC cells such as HepG2 and HuH7 550:598 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 1 26 theme sulfate 170:176 arg1 proteoglycan 178:189 a complex heparan sulfate proteoglycan 152:189 a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor 152:293 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 1 26 theme sulfate 170:176 arg1 Glypican 131:138 Glypican 3 131:140 Glypican 3 (GPC3) 131:147 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 3 27 theme biological 382:391 arg1 functions 393:401 numerous biological functions 373:401 numerous biological functions 373:401 GPC3 has numerous biological functions. 29345911 9 28 theme area 1154:1157 arg1 enlargement 1121:1131 the enlargement 1117:1131 the enlargement of HuH7 cell nuclear area 1117:1157 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 9 28 theme area 1154:1157 arg1 restructuration 1167:1181 the restructuration 1163:1181 the restructuration of adherent cell junctions 1163:1208 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 11 29 theme cell 1484:1487 arg1 proliferation 1489:1501 HCC cell proliferation 1480:1501 HCC cell proliferation 1480:1501 Overall, our data show that glycanation and convertase maturation are not required for sGPC3m to inhibit HCC cell proliferation. 29345911 1 30 theme GPI 283:285 arg1 anchor 288:293 a glycosylphosphatidylinositol (GPI) anchor 251:293 a glycosylphosphatidylinositol (GPI) anchor 251:293 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 7 31 theme doubling 1028:1035 arg1 time 1037:1040 their cell doubling time 1017:1040 their cell doubling time 1017:1040 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 6 32 theme post-translational 737:754 arg1 modifications 756:768 its post-translational modifications 733:768 its post-translational modifications 733:768 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 0 33 theme Glycanation 8:18 arg1 Role 0:3 Role 0:3 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 0:129 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 11 34 theme HCC 1480:1482 arg1 proliferation 1489:1501 HCC cell proliferation 1480:1501 HCC cell proliferation 1480:1501 Overall, our data show that glycanation and convertase maturation are not required for sGPC3m to inhibit HCC cell proliferation. 29345911 10 35 theme apoptosis 1260:1268 arg1 levels 1250:1255 the levels 1246:1255 the levels of apoptosis, cell division, and β-catenin 1246:1298 Unexpectedly, for both cell types, the levels of apoptosis, cell division, and β-catenin were not altered by sGPC3m, although growth inhibition was very efficient. 29345911 0 36 theme Maturation 35:44 arg1 Role 0:3 Role 0:3 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 0:129 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 4 37 from tissue 450:455 arg1 undetectable 421:432 undetectable 421:432 undetectable 421:432 Although GPC3 is undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). 29345911 4 38 theme liver 444:448 arg1 tissue 450:455 normal liver tissue 437:455 normal liver tissue 437:455 Although GPC3 is undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). 29345911 0 39 theme Convertase 24:33 arg1 Maturation 35:44 Convertase Maturation 24:44 Convertase Maturation 24:44 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 7 40 theme cell 1023:1026 arg1 time 1037:1040 their cell doubling time 1017:1040 their cell doubling time 1017:1040 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 6 41 theme GPI 818:820 arg1 site 850:853 convertase cleavage site 830:853 convertase cleavage site 830:853 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 6 41 theme GPI 818:820 arg1 anchor 822:827 its GPI anchor 814:827 its GPI anchor 814:827 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 10 42 theme cell 1271:1274 arg1 division 1276:1283 cell division 1271:1283 cell division 1271:1283 Unexpectedly, for both cell types, the levels of apoptosis, cell division, and β-catenin were not altered by sGPC3m, although growth inhibition was very efficient. 29345911 1 43 theme outer 211:215 arg1 surface 217:223 the outer surface 207:223 the outer surface of the plasma membrane 207:246 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 29345911 0 44 theme Soluble 49:55 arg1 Glypican-3 57:66 Soluble Glypican-3 49:66 Soluble Glypican-3 49:66 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 5 45 theme GPC3 649:652 arg1 form 641:644 a soluble form 631:644 a soluble form of GPC3 after lentiviral transduction 631:682 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 9 46 theme adherent 1186:1193 arg1 junctions 1200:1208 adherent cell junctions 1186:1208 adherent cell junctions 1186:1208 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 5 47 theme lentiviral 660:669 arg1 transduction 671:682 lentiviral transduction 660:682 lentiviral transduction 660:682 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 9 48 theme cell 1195:1198 arg1 junctions 1200:1208 adherent cell junctions 1186:1208 adherent cell junctions 1186:1208 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 7 49 theme pure 897:900 arg1 protein 909:915 The highly pure sGPC3m protein 886:915 The highly pure sGPC3m protein 886:915 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 6 50 theme some 725:728 arg1 role 717:720 the role 713:720 the role of some of its post-translational modifications 713:768 To obtain more insight into the role of some of its post-translational modifications, we designed a mutant GPC3, sGPC3m, without its GPI anchor, convertase cleavage site, and glycosaminoglycan chains. 29345911 4 51 from undetectable 421:432 arg1 tissue 450:455 normal liver tissue 437:455 normal liver tissue 437:455 Although GPC3 is undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). 29345911 5 52 theme HCC 567:569 arg1 HuH7 595:598 HuH7 595:598 HuH7 595:598 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 5 52 theme HCC 567:569 arg1 cells 571:575 HCC cells 567:575 HCC cells such as HepG2 and HuH7 567:598 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 5 52 theme HCC 567:569 arg1 HepG2 585:589 HepG2 585:589 HepG2 585:589 Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction. 29345911 8 53 theme cells 1077:1081 arg1 morphology 1058:1067 the morphology 1054:1067 the morphology of HuH7 cells but not that of HepG2 1054:1103 It changed the morphology of HuH7 cells but not that of HepG2. 29345911 7 54 theme HuH7 936:939 arg1 proliferation 956:968 HuH7 and HepG2 cell proliferation 936:968 HuH7 and HepG2 cell proliferation 936:968 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 7 55 theme significant 993:1003 arg1 increase 1005:1012 a significant increase 991:1012 a significant increase in their cell doubling time 991:1040 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 4 56 theme normal 437:442 arg1 tissue 450:455 normal liver tissue 437:455 normal liver tissue 437:455 Although GPC3 is undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). 29345911 7 57 theme cell 951:954 arg1 proliferation 956:968 HuH7 and HepG2 cell proliferation 936:968 HuH7 and HepG2 cell proliferation 936:968 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 7 58 theme sGPC3m 902:907 arg1 protein 909:915 The highly pure sGPC3m protein 886:915 The highly pure sGPC3m protein 886:915 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 11 59 theme convertase 1419:1428 arg1 maturation 1430:1439 convertase maturation 1419:1439 convertase maturation 1419:1439 Overall, our data show that glycanation and convertase maturation are not required for sGPC3m to inhibit HCC cell proliferation. 29345911 9 60 theme junctions 1200:1208 arg1 enlargement 1121:1131 the enlargement 1117:1131 the enlargement of HuH7 cell nuclear area 1117:1157 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 9 60 theme junctions 1200:1208 arg1 restructuration 1167:1181 the restructuration 1163:1181 the restructuration of adherent cell junctions 1163:1208 It induced the enlargement of HuH7 cell nuclear area and the restructuration of adherent cell junctions. 29345911 7 61 from increase 1005:1012 arg1 time 1037:1040 their cell doubling time 1017:1040 their cell doubling time 1017:1040 The highly pure sGPC3m protein strongly inhibited HuH7 and HepG2 cell proliferation in vitro and induced a significant increase in their cell doubling time. 29345911 0 62 theme Glypican-3 57:66 arg1 Glycanation 8:18 Glycanation 8:18 Glycanation 8:18 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 0 62 theme Glypican-3 57:66 arg1 Maturation 35:44 Convertase Maturation 24:44 Convertase Maturation 24:44 Role of Glycanation and Convertase Maturation of Soluble Glypican-3 in Inhibiting Proliferation of Hepatocellular Carcinoma Cells. 29345911 8 63 theme HuH7 1072:1075 arg1 cells 1077:1081 HuH7 cells 1072:1081 HuH7 cells 1072:1081 It changed the morphology of HuH7 cells but not that of HepG2. 29345911 1 64 theme plasma 232:237 arg1 membrane 239:246 the plasma membrane 228:246 the plasma membrane 228:246 Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. 19467646 9 0 theme glycosylated 1169:1180 arg1 pro-form 1182:1189 the glycosylated pro-form 1165:1189 the glycosylated pro-form of BDNF 1165:1197 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 5 1 theme cleavage 627:634 arg1 products 636:643 Proteolytic cleavage products 615:643 Proteolytic cleavage products 615:643 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 5 2 theme N-glycanase 716:726 arg1 F 728:728 N-glycanase F 716:728 N-glycanase F 716:728 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 11 3 theme BDNF 1524:1527 arg1 studies 1513:1519 future studies 1506:1519 future studies of BDNF and the Val66Met polymorphism 1506:1557 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 9 4 theme molecular 1130:1138 arg1 bands 1147:1151 the higher molecular weight bands 1119:1151 the higher molecular weight bands 1119:1151 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 0 5 from Identification 0:13 arg1 saliva 77:82 human saliva 71:82 human saliva 71:82 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 11 6 theme salivary 1383:1390 arg1 BDNF 1392:1395 salivary BDNF 1383:1395 salivary BDNF 1383:1395 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 2 7 theme Numerous 271:278 arg1 factors 287:293 Numerous growth factors 271:293 Numerous growth factors 271:293 Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. 19467646 1 8 theme central 241:247 arg1 cells 264:268 central and peripheral cells 241:268 central and peripheral cells 241:268 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 8 9 contain contained 1031:1039 arg1 sample 1024:1029 Not every sample 1014:1029 Not every sample 1014:1029 Not every sample contained all forms of BDNF. 19467646 8 9 contain contained 1031:1039 arg2 forms 1045:1049 all forms 1041:1049 all forms of BDNF 1041:1057 Not every sample contained all forms of BDNF. 19467646 9 10 theme BDNF 1194:1197 arg1 pro-form 1182:1189 the glycosylated pro-form 1165:1189 the glycosylated pro-form of BDNF 1165:1197 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 5 11 theme pooled 697:702 arg1 saliva 704:709 pooled saliva 697:709 pooled saliva 697:709 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 4 12 theme = 531:531 arg1 n 529:529 n = 30 529:534 n = 30 529:534 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 4 12 theme = 531:531 arg1 samples 520:526 unstimulated saliva samples 500:526 unstimulated saliva samples (n = 30) 500:535 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 10 13 theme Val66Met 1265:1272 arg1 SNP 1274:1276 the Val66Met SNP 1261:1276 the Val66Met SNP 1261:1276 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 7 14 from kDa 921:923 arg1 presence 871:878 the presence 867:878 the presence of immunoreactive bands at 14, 32 and 34 kDa 867:923 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 8 15 theme BDNF 1054:1057 arg1 forms 1045:1049 all forms 1041:1049 all forms of BDNF 1041:1057 Not every sample contained all forms of BDNF. 19467646 10 16 with subjects 1247:1254 arg1 SNP 1274:1276 the Val66Met SNP 1261:1276 the Val66Met SNP 1261:1276 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 7 17 theme immunoreactive 883:896 arg1 bands 898:902 immunoreactive bands 883:902 immunoreactive bands 883:902 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 9 18 theme higher 1123:1128 arg1 bands 1147:1151 the higher molecular weight bands 1119:1151 the higher molecular weight bands 1119:1151 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 9 19 with Treatment 1060:1068 arg1 N-glycanase 1075:1085 N-glycanase 1075:1085 N-glycanase 1075:1085 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 9 19 with Treatment 1060:1068 arg1 plasmin 1091:1097 plasmin 1091:1097 plasmin 1091:1097 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 11 20 theme Val66Met 1537:1544 arg1 polymorphism 1546:1557 the Val66Met polymorphism 1533:1557 the Val66Met polymorphism 1533:1557 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 11 21 from blood 1497:1501 arg1 studies 1513:1519 future studies 1506:1519 future studies of BDNF and the Val66Met polymorphism 1506:1557 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 4 22 theme unstimulated 500:511 arg1 n 529:529 n = 30 529:534 n = 30 529:534 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 4 22 theme unstimulated 500:511 arg1 samples 520:526 unstimulated saliva samples 500:526 unstimulated saliva samples (n = 30) 500:535 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 1 23 theme OBJECTIVE 85:93 arg1 factor 148:153 brain-derived neurotrophic factor 121:153 brain-derived neurotrophic factor (BDNF) 121:160 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 23 theme OBJECTIVE 85:93 arg1 factors 102:108 OBJECTIVE Growth factors 85:108 OBJECTIVE Growth factors 85:108 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 23 theme OBJECTIVE 85:93 arg1 polypeptides 167:178 polypeptides 167:178 polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells 167:268 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 4 24 theme BDNF 600:603 arg1 antibody 605:612 an anti-human BDNF antibody 586:612 an anti-human BDNF antibody 586:612 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 1 25 theme peripheral 253:262 arg1 cells 264:268 central and peripheral cells 241:268 central and peripheral cells 241:268 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 7 26 theme truncated 984:992 arg1 pro-form 994:1001 a truncated pro-form 982:1001 a truncated pro-form at 24 kDa 982:1011 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 0 27 link brain-derived 34:46 arg1 factor 61:66 pro- and mature brain-derived neurotrophic factor 18:66 pro- and mature brain-derived neurotrophic factor 18:66 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 1 28 theme Growth 95:100 arg1 factor 148:153 brain-derived neurotrophic factor 121:153 brain-derived neurotrophic factor (BDNF) 121:160 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 28 theme Growth 95:100 arg1 factors 102:108 OBJECTIVE Growth factors 85:108 OBJECTIVE Growth factors 85:108 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 28 theme Growth 95:100 arg1 polypeptides 167:178 polypeptides 167:178 polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells 167:268 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 2 29 theme wound 353:357 arg1 healing 359:365 wound healing 353:365 wound healing 353:365 Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. 19467646 1 30 theme cells 264:268 arg1 survival 218:225 survival 218:225 survival 218:225 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 30 theme cells 264:268 arg1 death 232:236 death 232:236 death 232:236 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 30 theme cells 264:268 arg1 maintenance 205:215 maintenance 205:215 maintenance 205:215 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 7 31 dep RESULTS 832:838 arg1 revealed 858:865 revealed 858:865 revealed 858:865 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 10 32 dep detected 1210:1217 arg1 compared 1279:1286 compared 1279:1286 compared to those without the polymorphism (chi(2) = 4.05; P < 0.05) 1279:1346 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 2 33 theme epithelium 395:404 arg1 maintenance 371:381 maintenance 371:381 maintenance 371:381 Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. 19467646 2 33 theme epithelium 395:404 arg1 healing 359:365 wound healing 353:365 wound healing 353:365 Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. 19467646 0 34 theme pro- 18:21 arg1 factor 61:66 pro- and mature brain-derived neurotrophic factor 18:66 pro- and mature brain-derived neurotrophic factor 18:66 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 2 35 theme growth 280:285 arg1 factors 287:293 Numerous growth factors 271:293 Numerous growth factors 271:293 Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. 19467646 2 36 theme oral 390:393 arg1 epithelium 395:404 the oral epithelium 386:404 the oral epithelium 386:404 Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. 19467646 0 37 theme brain-derived 34:46 arg1 factor 61:66 pro- and mature brain-derived neurotrophic factor 18:66 pro- and mature brain-derived neurotrophic factor 18:66 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 4 38 theme saliva 513:518 arg1 n 529:529 n = 30 529:534 n = 30 529:534 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 4 38 theme saliva 513:518 arg1 samples 520:526 unstimulated saliva samples 500:526 unstimulated saliva samples (n = 30) 500:535 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 10 39 theme P 1338:1338 arg1 < 1340:1340 P < 0.05 1338:1345 P < 0.05 1338:1345 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 5 40 with incubation 683:692 arg1 plasmin 734:740 plasmin 734:740 plasmin 734:740 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 5 40 with incubation 683:692 arg1 F 728:728 N-glycanase F 716:728 N-glycanase F 716:728 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 4 41 theme anti-human 589:598 arg1 antibody 605:612 an anti-human BDNF antibody 586:612 an anti-human BDNF antibody 586:612 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 11 42 dep CONCLUSIONS 1349:1359 arg1 requires 1403:1410 requires 1403:1410 requires elucidation 1403:1422 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 0 43 theme mature 27:32 arg1 factor 61:66 pro- and mature brain-derived neurotrophic factor 18:66 pro- and mature brain-derived neurotrophic factor 18:66 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 11 44 theme BDNF 1392:1395 arg1 function 1371:1378 the function 1367:1378 the function of salivary BDNF 1367:1395 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 6 45 theme nucleotide 801:810 arg1 SNP 826:828 SNP 826:828 SNP 826:828 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 6 45 theme nucleotide 801:810 arg1 polymorphism 812:823 the BDNF Val66Met single nucleotide polymorphism 776:823 the BDNF Val66Met single nucleotide polymorphism (SNP) 776:829 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 7 46 theme mature 943:948 arg1 mBDNF 951:955 mature (mBDNF) 943:956 mature (mBDNF) 943:956 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 3 47 theme study 423:427 arg1 aim 411:413 The aim 407:413 The aim of this study 407:427 The aim of this study was to determine if BDNF is also found in human saliva. 19467646 0 48 theme factor 61:66 arg1 Identification 0:13 Identification 0:13 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 0:83 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 7 49 from kDa 1009:1011 arg1 pro-form 994:1001 a truncated pro-form 982:1001 a truncated pro-form at 24 kDa 982:1011 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 7 49 from kDa 1009:1011 arg1 mBDNF 951:955 mature (mBDNF) 943:956 mature (mBDNF) 943:956 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 7 49 from kDa 1009:1011 arg1 proBDNF 962:968 proBDNF 962:968 proBDNF 962:968 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 6 50 theme single 794:799 arg1 SNP 826:828 SNP 826:828 SNP 826:828 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 6 50 theme single 794:799 arg1 polymorphism 812:823 the BDNF Val66Met single nucleotide polymorphism 776:823 the BDNF Val66Met single nucleotide polymorphism (SNP) 776:829 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 9 51 gly glycosylated 1169:1180 arg1 pro-form 1182:1189 the glycosylated pro-form 1165:1189 the glycosylated pro-form of BDNF 1165:1197 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 9 51 gly glycosylated 1169:1180 arg1 BDNF 1194:1197 BDNF 1194:1197 BDNF 1194:1197 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 0 52 theme neurotrophic 48:59 arg1 factor 61:66 pro- and mature brain-derived neurotrophic factor 18:66 pro- and mature brain-derived neurotrophic factor 18:66 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 9 53 theme weight 1140:1145 arg1 bands 1147:1151 the higher molecular weight bands 1119:1151 the higher molecular weight bands 1119:1151 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 6 54 theme Val66Met 785:792 arg1 SNP 826:828 SNP 826:828 SNP 826:828 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 6 54 theme Val66Met 785:792 arg1 polymorphism 812:823 the BDNF Val66Met single nucleotide polymorphism 776:823 the BDNF Val66Met single nucleotide polymorphism (SNP) 776:829 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 6 55 theme BDNF 780:783 arg1 SNP 826:828 SNP 826:828 SNP 826:828 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 6 55 theme BDNF 780:783 arg1 polymorphism 812:823 the BDNF Val66Met single nucleotide polymorphism 776:823 the BDNF Val66Met single nucleotide polymorphism (SNP) 776:829 Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP). 19467646 3 56 located found 462:466 arg1 saliva 477:482 human saliva 471:482 human saliva 471:482 The aim of this study was to determine if BDNF is also found in human saliva. 19467646 3 56 located found 462:466 arg2 BDNF 449:452 BDNF 449:452 BDNF 449:452 The aim of this study was to determine if BDNF is also found in human saliva. 19467646 1 57 theme brain-derived 121:133 arg1 BDNF 156:159 BDNF 156:159 BDNF 156:159 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 57 theme brain-derived 121:133 arg1 factor 148:153 brain-derived neurotrophic factor 121:153 brain-derived neurotrophic factor (BDNF) 121:160 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 5 58 theme Proteolytic 615:625 arg1 products 636:643 Proteolytic cleavage products 615:643 Proteolytic cleavage products 615:643 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 4 59 dep analyzed 542:549 arg1 Whole 493:497 Whole 493:497 Whole 493:497 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 1 60 theme neurotrophic 135:146 arg1 BDNF 156:159 BDNF 156:159 BDNF 156:159 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 60 theme neurotrophic 135:146 arg1 factor 148:153 brain-derived neurotrophic factor 121:153 brain-derived neurotrophic factor (BDNF) 121:160 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 61 link brain-derived 121:133 arg1 BDNF 156:159 BDNF 156:159 BDNF 156:159 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 1 61 link brain-derived 121:133 arg1 factor 148:153 brain-derived neurotrophic factor 121:153 brain-derived neurotrophic factor (BDNF) 121:160 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 10 62 located detected 1210:1217 arg1 subjects 1247:1254 subjects 1247:1254 subjects with the Val66Met SNP 1247:1276 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 10 62 located detected 1210:1217 arg2 mBDNF 1200:1204 mBDNF 1200:1204 mBDNF 1200:1204 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 1 63 dep maintenance 205:215 arg1 the 201:203 the 201:203 the 201:203 OBJECTIVE Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. 19467646 10 64 dep chi 1323:1325 arg1 < 1340:1340 P < 0.05 1338:1345 P < 0.05 1338:1345 mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05). 19467646 7 65 theme bands 898:902 arg1 presence 871:878 the presence 867:878 the presence of immunoreactive bands at 14, 32 and 34 kDa 867:923 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 0 66 theme human 71:75 arg1 saliva 77:82 human saliva 71:82 human saliva 71:82 Identification of pro- and mature brain-derived neurotrophic factor in human saliva. 19467646 4 67 theme Western 567:573 arg1 blot 575:578 Western blot 567:578 Western blot 567:578 METHODS Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. 19467646 9 68 theme bands 1147:1151 arg1 size 1111:1114 the size 1107:1114 the size of the higher molecular weight bands 1107:1151 Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. 19467646 11 69 theme polymorphism 1546:1557 arg1 studies 1513:1519 future studies 1506:1519 future studies of BDNF and the Val66Met polymorphism 1506:1557 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 19467646 5 70 theme saliva 704:709 arg1 incubation 683:692 the incubation 679:692 the incubation of pooled saliva with N-glycanase F and plasmin 679:740 Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. 19467646 7 71 attach presence 871:878 arg1 kDa 921:923 14, 32 and 34 kDa 907:923 14, 32 and 34 kDa 907:923 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 7 71 attach presence 871:878 arg2 bands 898:902 immunoreactive bands 883:902 immunoreactive bands 883:902 RESULTS These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. 19467646 3 72 theme human 471:475 arg1 saliva 477:482 human saliva 471:482 human saliva 471:482 The aim of this study was to determine if BDNF is also found in human saliva. 19467646 11 73 theme future 1506:1511 arg1 studies 1513:1519 future studies 1506:1519 future studies of BDNF and the Val66Met polymorphism 1506:1557 CONCLUSIONS While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism. 22351761 4 0 theme HS 955:956 arg1 assembly 943:950 assembly 943:950 assembly of HS 943:956 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 6 1 theme attachment 1186:1195 arg1 domain 1197:1202 the HS attachment domain 1179:1202 the HS attachment domain 1179:1202 We expressed and crystallized N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain. 22351761 3 2 theme coil 753:756 arg1 conformation 758:769 a random coil conformation 744:769 a random coil conformation 744:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 1 3 theme fibroblast 255:264 arg1 factor 273:278 fibroblast growth factor 255:278 fibroblast growth factor 255:278 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 9 4 theme structural 1750:1759 arg1 knowledge 1761:1769 the structural knowledge 1746:1769 the structural knowledge of glypicans for one α-helix and two long loops 1746:1817 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 5 theme glypicans 1774:1782 arg1 knowledge 1761:1769 the structural knowledge 1746:1769 the structural knowledge of glypicans for one α-helix and two long loops 1746:1817 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 7 6 theme crystal 1209:1215 arg1 structure 1217:1225 The crystal structure 1205:1225 The crystal structure of glypican-1 1205:1239 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 2 7 theme Loss-of-function 291:306 arg1 mutations 308:316 Loss-of-function mutations 291:316 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes 291:388 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 1 8 theme factor 273:278 arg1 signaling 280:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 8 9 theme electron 1356:1363 arg1 density 1365:1371 No additional electron density 1342:1371 No additional electron density 1342:1371 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 7 10 theme glypican-1 1295:1304 arg1 protein 1311:1317 selenomethionine-labeled glypican-1 core protein 1270:1317 selenomethionine-labeled glypican-1 core protein lacking the HS domain 1270:1339 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 1 11 theme proteoglycans 180:192 arg1 Glypicans 141:149 Glypicans 141:149 Glypicans 141:149 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 1 11 theme proteoglycans 180:192 arg1 family 157:162 a family 155:162 a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 155:288 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 10 12 theme vertebrate 1875:1884 arg1 glypican-1 1886:1895 vertebrate glypican-1 1875:1895 vertebrate glypican-1 1875:1895 Importantly, the loops are evolutionarily conserved in vertebrate glypican-1, and one of them is involved in glycosaminoglycan class determination. 22351761 2 13 theme core 330:333 arg1 proteins 335:342 glypican core proteins 321:342 glypican core proteins 321:342 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 9 14 theme protein 1575:1581 arg1 structure 1525:1533 The crystal structure 1513:1533 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, 1513:1645 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 15 theme human 1553:1557 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 15 theme human 1553:1557 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 0 16 theme vertebrate 118:127 arg1 glypican-1 129:138 vertebrate glypican-1 118:138 vertebrate glypican-1 118:138 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 4 17 from factors 903:909 arg1 domain 928:933 the α-helical domain 914:933 the α-helical domain 914:933 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 9 18 theme first 1597:1601 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 18 theme first 1597:1601 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 8 19 theme HS 1428:1429 arg1 domain 1442:1447 the HS attachment domain 1424:1447 the HS attachment domain 1424:1447 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 2 20 from mutations 308:316 arg1 enzymes 382:388 glycosaminoglycan-synthesizing enzymes 351:388 glycosaminoglycan-synthesizing enzymes 351:388 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 2 20 from mutations 308:316 arg1 proteins 335:342 glypican core proteins 321:342 glypican core proteins 321:342 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 6 21 theme N-glycosylated 1098:1111 arg1 glypican-1 1119:1128 N-glycosylated human glypican-1 1098:1128 N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain 1098:1202 We expressed and crystallized N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain. 22351761 10 22 theme them 1909:1912 arg1 one 1902:1904 one 1902:1904 one 1902:1904 Importantly, the loops are evolutionarily conserved in vertebrate glypican-1, and one of them is involved in glycosaminoglycan class determination. 22351761 10 22 theme them 1909:1912 arg1 them 1909:1912 them 1909:1912 them 1909:1912 Importantly, the loops are evolutionarily conserved in vertebrate glypican-1, and one of them is involved in glycosaminoglycan class determination. 22351761 5 23 theme brain 985:989 arg1 development 991:1001 brain development 985:1001 brain development 985:1001 Glypican-1 is involved in brain development and is one of six members of the vertebrate family of glypicans. 22351761 3 24 contain containing 572:581 arg1 domain 565:570 a stable α-helical domain 546:570 a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 546:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 3 24 contain containing 572:581 arg2 residues 600:607 14 conserved Cys residues 583:607 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 583:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 0 25 theme loops 84:88 arg1 structure 67:75 structure 67:75 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 0:139 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 5 26 theme vertebrate 1036:1045 arg1 family 1047:1052 the vertebrate family 1032:1052 the vertebrate family of glypicans 1032:1065 Glypican-1 is involved in brain development and is one of six members of the vertebrate family of glypicans. 22351761 4 27 theme glycosaminoglycan 848:864 arg1 sulfate 878:884 the glycosaminoglycan chondroitin sulfate 844:884 the glycosaminoglycan chondroitin sulfate 844:884 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 7 28 theme protein 1311:1317 arg1 crystals 1258:1265 crystals 1258:1265 crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain 1258:1339 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 9 29 theme complete 1659:1666 arg1 arrangement 1683:1693 the complete disulfide bond arrangement 1655:1693 the complete disulfide bond arrangement of the conserved Cys residues 1655:1723 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 8 30 theme protein 1476:1482 arg1 species 1484:1490 the two protein species 1468:1490 the two protein species 1468:1490 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 0 31 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 0:139 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 9 32 theme bond 1678:1681 arg1 arrangement 1683:1693 the complete disulfide bond arrangement 1655:1693 the complete disulfide bond arrangement of the conserved Cys residues 1655:1723 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 2 33 theme shaping 484:490 arg1 development 499:509 shaping animal development 484:509 shaping animal development 484:509 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 4 34 theme exclusive 822:830 arg1 addition 832:839 almost exclusive addition 815:839 almost exclusive addition of the glycosaminoglycan chondroitin sulfate 815:884 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 3 35 theme core 521:524 arg1 proteins 526:533 Glypican core proteins 512:533 Glypican core proteins 512:533 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 9 36 theme N-glycosylated 1538:1551 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 36 theme N-glycosylated 1538:1551 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 0 37 theme human 36:40 arg1 protein 58:64 N-glycosylated human glypican-1 core protein 21:64 N-glycosylated human glypican-1 core protein 21:64 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 9 38 from structure 1525:1533 arg1 Å 1590:1590 2.5 Å 1586:1590 2.5 Å 1586:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 39 theme conserved 1702:1710 arg1 residues 1716:1723 the conserved Cys residues 1698:1723 the conserved Cys residues 1698:1723 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 40 from Å 1590:1590 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 40 from Å 1590:1590 arg1 structure 1525:1533 The crystal structure 1513:1533 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, 1513:1645 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 40 from Å 1590:1590 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 10 41 theme glycosaminoglycan 1929:1945 arg1 determination 1953:1965 glycosaminoglycan class determination 1929:1965 glycosaminoglycan class determination 1929:1965 Importantly, the loops are evolutionarily conserved in vertebrate glypican-1, and one of them is involved in glycosaminoglycan class determination. 22351761 0 42 theme core 53:56 arg1 protein 58:64 N-glycosylated human glypican-1 core protein 21:64 N-glycosylated human glypican-1 core protein 21:64 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 0 43 gly N-glycosylated 21:34 arg1 protein 58:64 N-glycosylated human glypican-1 core protein 21:64 N-glycosylated human glypican-1 core protein 21:64 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 3 44 theme random 746:751 arg1 conformation 758:769 a random coil conformation 744:769 a random coil conformation 744:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 4 45 theme sulfate 878:884 arg1 addition 832:839 almost exclusive addition 815:839 almost exclusive addition of the glycosaminoglycan chondroitin sulfate 815:884 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 1 46 theme bone 223:226 arg1 protein 242:248 bone morphogenetic protein 223:248 bone morphogenetic protein 223:248 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 3 47 theme Cys 596:598 arg1 residues 600:607 14 conserved Cys residues 583:607 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 583:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 3 48 theme Glypican 512:519 arg1 proteins 526:533 Glypican core proteins 512:533 Glypican core proteins 512:533 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 9 49 gly N-glycosylated 1538:1551 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 49 gly N-glycosylated 1538:1551 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 2 50 theme glypican 409:416 arg1 proteins 423:430 glypican core proteins 409:430 glypican core proteins 409:430 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 9 51 theme glypican-1 1559:1568 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 51 theme glypican-1 1559:1568 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 1 52 theme protein 242:248 arg1 signaling 280:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 0 53 theme protein 58:64 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 0:139 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 2 54 from important 471:479 arg1 development 499:509 shaping animal development 484:509 shaping animal development 484:509 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 3 55 theme α-helical 555:563 arg1 domain 565:570 a stable α-helical domain 546:570 a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 546:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 9 56 theme crystal 1517:1523 arg1 structure 1525:1533 The crystal structure 1513:1533 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, 1513:1645 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 6 57 theme HS 1183:1184 arg1 domain 1197:1202 the HS attachment domain 1179:1202 the HS attachment domain 1179:1202 We expressed and crystallized N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain. 22351761 1 58 theme growth 266:271 arg1 factor 273:278 fibroblast growth factor 255:278 fibroblast growth factor 255:278 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 3 59 theme conserved 586:594 arg1 residues 600:607 14 conserved Cys residues 583:607 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 583:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 1 60 theme cell-surface 167:178 arg1 proteoglycans 180:192 cell-surface proteoglycans 167:192 cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 167:288 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 4 61 theme α-helical 918:926 arg1 domain 928:933 the α-helical domain 914:933 the α-helical domain 914:933 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 8 62 theme additional 1345:1354 arg1 density 1365:1371 No additional electron density 1342:1371 No additional electron density 1342:1371 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 3 63 theme glycosaminoglycan 623:639 arg1 domain 652:657 a glycosaminoglycan attachment domain 621:657 a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 621:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 9 64 theme core 1570:1573 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 64 theme core 1570:1573 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 65 theme residues 1716:1723 arg1 arrangement 1683:1693 the complete disulfide bond arrangement 1655:1693 the complete disulfide bond arrangement of the conserved Cys residues 1655:1723 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 2 66 theme glypican 321:328 arg1 proteins 335:342 glypican core proteins 321:342 glypican core proteins 321:342 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 1 67 theme Wnt 208:210 arg1 signaling 280:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 3 68 theme attachment 641:650 arg1 domain 652:657 a glycosaminoglycan attachment domain 621:657 a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 621:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 9 69 theme glypican 1637:1644 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 69 theme glypican 1637:1644 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 2 70 theme glycosaminoglycan-synthesizing 351:380 arg1 enzymes 382:388 glycosaminoglycan-synthesizing enzymes 351:388 glycosaminoglycan-synthesizing enzymes 351:388 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 1 71 theme hedgehog 213:220 arg1 signaling 280:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling 208:288 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 9 72 theme long 1808:1811 arg1 loops 1813:1817 two long loops 1804:1817 two long loops 1804:1817 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 8 73 theme glypican-1 1402:1411 arg1 crystals 1390:1397 crystals 1390:1397 crystals of glypican-1 containing the HS attachment domain 1390:1447 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 3 74 theme heparan 701:707 arg1 HS 718:719 HS 718:719 HS 718:719 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 3 74 theme heparan 701:707 arg1 sulfate 709:715 heparan sulfate 701:715 heparan sulfate (HS) 701:720 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 9 75 theme crystal 1603:1609 arg1 protein 1575:1581 N-glycosylated human glypican-1 core protein 1538:1581 N-glycosylated human glypican-1 core protein at 2.5 Å 1538:1590 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 9 75 theme crystal 1603:1609 arg1 structure 1611:1619 the first crystal structure 1593:1619 the first crystal structure of a vertebrate glypican 1593:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 6 76 theme human 1113:1117 arg1 glypican-1 1119:1128 N-glycosylated human glypican-1 1098:1128 N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain 1098:1202 We expressed and crystallized N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain. 22351761 8 77 contain containing 1413:1422 arg1 glypican-1 1402:1411 glypican-1 1402:1411 glypican-1 containing the HS attachment domain 1402:1447 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 8 77 contain containing 1413:1422 arg2 domain 1442:1447 the HS attachment domain 1424:1447 the HS attachment domain 1424:1447 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 4 78 theme domain 797:802 arg1 Removal 772:778 Removal 772:778 Removal of the α-helical domain 772:802 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 4 79 theme α-helical 787:795 arg1 domain 797:802 the α-helical domain 783:802 the α-helical domain 783:802 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 8 80 theme attachment 1431:1440 arg1 domain 1442:1447 the HS attachment domain 1424:1447 the HS attachment domain 1424:1447 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 7 81 theme HS 1331:1332 arg1 domain 1334:1339 the HS domain 1327:1339 the HS domain 1327:1339 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 9 82 theme vertebrate 1626:1635 arg1 glypican 1637:1644 a vertebrate glypican 1624:1644 a vertebrate glypican 1624:1644 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 2 83 from development 499:509 arg1 important 471:479 important 471:479 important 471:479 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 8 84 theme CD 1454:1455 arg1 spectra 1457:1463 CD spectra 1454:1463 CD spectra of the two protein species 1454:1490 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 7 85 theme core 1306:1309 arg1 protein 1311:1317 selenomethionine-labeled glypican-1 core protein 1270:1317 selenomethionine-labeled glypican-1 core protein lacking the HS domain 1270:1339 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 5 86 theme family 1047:1052 arg1 members 1021:1027 six members 1017:1027 six members of the vertebrate family of glypicans 1017:1065 Glypican-1 is involved in brain development and is one of six members of the vertebrate family of glypicans. 22351761 6 87 dep HS 1138:1139 arg1 glypican-1 1160:1169 glypican-1 1160:1169 glypican-1 1160:1169 We expressed and crystallized N-glycosylated human glypican-1 lacking HS and N-glycosylated glypican-1 lacking the HS attachment domain. 22351761 7 88 theme selenomethionine-labeled 1270:1293 arg1 protein 1311:1317 selenomethionine-labeled glypican-1 core protein 1270:1317 selenomethionine-labeled glypican-1 core protein lacking the HS domain 1270:1339 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 9 89 theme disulfide 1668:1676 arg1 arrangement 1683:1693 the complete disulfide bond arrangement 1655:1693 the complete disulfide bond arrangement of the conserved Cys residues 1655:1723 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 2 90 theme animal 492:497 arg1 development 499:509 shaping animal development 484:509 shaping animal development 484:509 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 2 91 theme core 418:421 arg1 proteins 423:430 glypican core proteins 409:430 glypican core proteins 409:430 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 0 92 theme N-glycosylated 21:34 arg1 protein 58:64 N-glycosylated human glypican-1 core protein 21:64 N-glycosylated human glypican-1 core protein 21:64 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 5 93 theme members 1021:1027 arg1 members 1021:1027 six members 1017:1027 six members of the vertebrate family of glypicans 1017:1065 Glypican-1 is involved in brain development and is one of six members of the vertebrate family of glypicans. 22351761 5 93 theme members 1021:1027 arg1 one 1010:1012 one 1010:1012 one 1010:1012 Glypican-1 is involved in brain development and is one of six members of the vertebrate family of glypicans. 22351761 8 94 theme species 1484:1490 arg1 spectra 1457:1463 CD spectra 1454:1463 CD spectra of the two protein species 1454:1490 No additional electron density was observed for crystals of glypican-1 containing the HS attachment domain, and CD spectra of the two protein species were highly similar. 22351761 0 95 theme glypican-1 42:51 arg1 protein 58:64 N-glycosylated human glypican-1 core protein 21:64 N-glycosylated human glypican-1 core protein 21:64 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 22351761 10 96 theme class 1947:1951 arg1 determination 1953:1965 glycosaminoglycan class determination 1929:1965 glycosaminoglycan class determination 1929:1965 Importantly, the loops are evolutionarily conserved in vertebrate glypican-1, and one of them is involved in glycosaminoglycan class determination. 22351761 2 97 theme glycosaminoglycan 442:458 arg1 chains 460:465 their glycosaminoglycan chains 436:465 their glycosaminoglycan chains 436:465 Loss-of-function mutations in glypican core proteins and in glycosaminoglycan-synthesizing enzymes have revealed that glypican core proteins and their glycosaminoglycan chains are important in shaping animal development. 22351761 7 98 theme glypican-1 1230:1239 arg1 structure 1217:1225 The crystal structure 1205:1225 The crystal structure of glypican-1 1205:1239 The crystal structure of glypican-1 was solved using crystals of selenomethionine-labeled glypican-1 core protein lacking the HS domain. 22351761 9 99 theme Cys 1712:1714 arg1 residues 1716:1723 the conserved Cys residues 1698:1723 the conserved Cys residues 1698:1723 The crystal structure of N-glycosylated human glypican-1 core protein at 2.5 Å, the first crystal structure of a vertebrate glypican, reveals the complete disulfide bond arrangement of the conserved Cys residues, and it also extends the structural knowledge of glypicans for one α-helix and two long loops. 22351761 3 100 theme stable 548:553 arg1 domain 565:570 a stable α-helical domain 546:570 a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation 546:769 Glypican core proteins consist of a stable α-helical domain containing 14 conserved Cys residues followed by a glycosaminoglycan attachment domain that becomes exclusively substituted with heparan sulfate (HS) and presumably adopts a random coil conformation. 22351761 5 101 theme glypicans 1057:1065 arg1 family 1047:1052 the vertebrate family 1032:1052 the vertebrate family of glypicans 1032:1065 Glypican-1 is involved in brain development and is one of six members of the vertebrate family of glypicans. 22351761 4 102 theme chondroitin 866:876 arg1 sulfate 878:884 the glycosaminoglycan chondroitin sulfate 844:884 the glycosaminoglycan chondroitin sulfate 844:884 Removal of the α-helical domain results in almost exclusive addition of the glycosaminoglycan chondroitin sulfate, suggesting that factors in the α-helical domain promote assembly of HS. 22351761 1 103 theme morphogenetic 228:240 arg1 protein 242:248 bone morphogenetic protein 223:248 bone morphogenetic protein 223:248 Glypicans are a family of cell-surface proteoglycans that regulate Wnt, hedgehog, bone morphogenetic protein, and fibroblast growth factor signaling. 22351761 0 104 dep structure 8:16 arg1 structure 67:75 structure 67:75 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 0:139 Crystal structure of N-glycosylated human glypican-1 core protein: structure of two loops evolutionarily conserved in vertebrate glypican-1. 16912035 7 0 theme enhanced 1178:1185 arg1 susceptibility 1187:1200 an enhanced susceptibility 1175:1200 an enhanced susceptibility to furin cleavage (A443T) 1175:1226 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 1 theme endoplasmic 1269:1279 arg1 C679X 1292:1296 C679X 1292:1296 C679X 1292:1296 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 1 theme endoplasmic 1269:1279 arg1 reticulum 1281:1289 the endoplasmic reticulum 1265:1289 the endoplasmic reticulum (C679X) 1265:1297 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 6 2 theme partial 955:961 arg1 loss 963:966 total or partial loss 946:966 total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow 946:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 10 3 theme proteinase 1571:1580 arg1 lifetime 1554:1561 the lifetime 1550:1561 the lifetime of this proteinase 1550:1580 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 10 3 theme proteinase 1571:1580 arg1 ability 1590:1596 its ability to degrade the cell-surface LDL receptor 1586:1637 its ability to degrade the cell-surface LDL receptor 1586:1637 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 6 4 from arrow 1025:1029 arg1 loss 963:966 total or partial loss 946:966 total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow 946:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 2 5 theme form 325:328 arg1 development 299:309 the development 295:309 the development of a dominant form of familial hyper- or hypocholesterolemia 295:370 Some of its natural mutations have been genetically associated with the development of a dominant form of familial hyper- or hypocholesterolemia. 16912035 6 6 theme gain-of-function 848:863 arg1 mutations 865:873 the natural gain-of-function mutations 836:873 the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia 836:934 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 6 theme gain-of-function 848:863 arg1 F216L 882:886 F216L 882:886 F216L 882:886 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 6 theme gain-of-function 848:863 arg1 D374Y 893:897 D374Y 893:897 D374Y 893:897 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 6 theme gain-of-function 848:863 arg1 R218S 875:879 R218S 875:879 R218S 875:879 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 3 7 theme density 499:505 arg1 LDL 520:522 LDL 520:522 LDL 520:522 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 3 7 theme density 499:505 arg1 lipoprotein 507:517 low density lipoprotein 495:517 the low density lipoprotein (LDL) receptor 491:532 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 8 8 theme cleaved 1366:1372 arg1 forms 1374:1378 both native and furin-like cleaved forms 1339:1378 both native and furin-like cleaved forms of PCSK9 1339:1387 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 6 9 theme total 946:950 arg1 loss 963:966 total or partial loss 946:966 total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow 946:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 7 10 theme localization 1158:1169 arg1 lack 1111:1114 the lack 1107:1114 the lack of trans-Golgi network/recycling endosome localization 1107:1169 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 6 11 theme natural 840:846 arg1 mutations 865:873 the natural gain-of-function mutations 836:873 the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia 836:934 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 11 theme natural 840:846 arg1 F216L 882:886 F216L 882:886 F216L 882:886 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 11 theme natural 840:846 arg1 D374Y 893:897 D374Y 893:897 D374Y 893:897 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 11 theme natural 840:846 arg1 R218S 875:879 R218S 875:879 R218S 875:879 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 3 12 theme lipoprotein 507:517 arg1 receptor 525:532 the low density lipoprotein (LDL) receptor 491:532 the low density lipoprotein (LDL) receptor 491:532 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 5 13 theme family 809:814 arg1 members 791:797 the other members 781:797 the other members of the PC family 781:814 We also examined the susceptibility of PCSK9 to proteolytic cleavage by the other members of the PC family. 16912035 7 14 theme endosome 1149:1156 arg1 localization 1158:1169 trans-Golgi network/recycling endosome localization 1119:1169 trans-Golgi network/recycling endosome localization 1119:1169 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 10 15 theme LDL 1626:1628 arg1 receptor 1630:1637 the cell-surface LDL receptor 1609:1637 the cell-surface LDL receptor 1609:1637 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 7 16 dep mutations 1066:1074 arg1 A443T 1076:1080 A443T 1076:1080 A443T 1076:1080 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 16 dep mutations 1066:1074 arg1 mutations 1066:1074 the loss-of-function mutations 1045:1074 the loss-of-function mutations A443T and C679X 1045:1090 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 16 dep mutations 1066:1074 arg1 C679X 1086:1090 C679X 1086:1090 C679X 1086:1090 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 1 17 theme ninth 172:176 arg1 member 178:183 the ninth member 168:183 the ninth member of the proprotein convertase (PC) family 168:224 PCSK9 is the ninth member of the proprotein convertase (PC) family. 16912035 1 17 theme ninth 172:176 arg1 PCSK9 159:163 PCSK9 159:163 PCSK9 159:163 PCSK9 is the ninth member of the proprotein convertase (PC) family. 16912035 5 18 theme PCSK9 748:752 arg1 susceptibility 730:743 the susceptibility 726:743 the susceptibility of PCSK9 to proteolytic cleavage by the other members of the PC family 726:814 We also examined the susceptibility of PCSK9 to proteolytic cleavage by the other members of the PC family. 16912035 0 19 theme functional 76:85 arg1 consequences 87:98 functional consequences 76:98 functional consequences of natural mutations and post-translational modifications 76:156 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 6 20 dep mutations 865:873 arg1 mutations 865:873 the natural gain-of-function mutations 836:873 the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia 836:934 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 20 dep mutations 865:873 arg1 F216L 882:886 F216L 882:886 F216L 882:886 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 20 dep mutations 865:873 arg1 D374Y 893:897 D374Y 893:897 D374Y 893:897 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 20 dep mutations 865:873 arg1 R218S 875:879 R218S 875:879 R218S 875:879 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 2 21 theme natural 239:245 arg1 mutations 247:255 its natural mutations 235:255 its natural mutations 235:255 Some of its natural mutations have been genetically associated with the development of a dominant form of familial hyper- or hypocholesterolemia. 16912035 0 22 theme proprotein 4:13 arg1 convertase 15:24 proprotein convertase 4:24 The proprotein convertase (PC) PCSK9 0:35 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 0 22 theme proprotein 4:13 arg1 PC 27:28 PC 27:28 PC 27:28 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 9 23 theme basic 1481:1485 arg1 convertases 1498:1508 the basic amino acid convertases 1477:1508 the basic amino acid convertases furin and PC5/6A 1477:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 23 theme basic 1481:1485 arg1 PC5/6A 1520:1525 PC5/6A 1520:1525 PC5/6A 1520:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 23 theme basic 1481:1485 arg1 furin 1510:1514 furin 1510:1514 furin 1510:1514 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 24 theme amino 1487:1491 arg1 convertases 1498:1508 the basic amino acid convertases 1477:1508 the basic amino acid convertases furin and PC5/6A 1477:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 24 theme amino 1487:1491 arg1 PC5/6A 1520:1525 PC5/6A 1520:1525 PC5/6A 1520:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 24 theme amino 1487:1491 arg1 furin 1510:1514 furin 1510:1514 furin 1510:1514 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 8 25 from presence 1327:1334 arg1 plasma 1410:1415 circulating human plasma 1392:1415 circulating human plasma 1392:1415 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 2 26 theme hypocholesterolemia 352:370 arg1 form 325:328 a dominant form 314:328 a dominant form of familial hyper- or hypocholesterolemia 314:370 Some of its natural mutations have been genetically associated with the development of a dominant form of familial hyper- or hypocholesterolemia. 16912035 7 27 theme network/recycling 1131:1147 arg1 localization 1158:1169 trans-Golgi network/recycling endosome localization 1119:1169 trans-Golgi network/recycling endosome localization 1119:1169 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 4 28 from Tyr38 702:706 arg1 prosegment 688:697 its prosegment 684:697 its prosegment at Tyr38 684:706 We analyzed the post-translational modifications of PCSK9 and show that it is sulfated within its prosegment at Tyr38. 16912035 3 29 theme receptor 525:532 arg1 degradation 476:486 the intracellular degradation 458:486 the intracellular degradation of the low density lipoprotein (LDL) receptor 458:532 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 3 30 theme PCSK9 406:410 arg1 action 396:401 action 396:401 action of PCSK9 396:410 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 10 31 theme circulating 1673:1683 arg1 cholesterol 1689:1699 circulating LDL cholesterol 1673:1699 circulating LDL cholesterol 1673:1699 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 0 32 theme natural 103:109 arg1 mutations 111:119 natural mutations 103:119 natural mutations 103:119 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 10 33 theme cholesterol 1689:1699 arg1 levels 1663:1668 the levels 1659:1668 the levels of circulating LDL cholesterol 1659:1699 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 7 34 theme loss-of-function 1049:1064 arg1 A443T 1076:1080 A443T 1076:1080 A443T 1076:1080 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 34 theme loss-of-function 1049:1064 arg1 mutations 1066:1074 the loss-of-function mutations 1045:1074 the loss-of-function mutations A443T and C679X 1045:1090 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 34 theme loss-of-function 1049:1064 arg1 C679X 1086:1090 C679X 1086:1090 C679X 1086:1090 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 6 35 theme RFHR218 1008:1014 arg1 arrow 1025:1029 the motif RFHR218 downward arrow 998:1029 the motif RFHR218 downward arrow 998:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 0 36 theme convertase 15:24 arg1 PCSK9 31:35 The proprotein convertase (PC) PCSK9 0:35 The proprotein convertase (PC) PCSK9 0:35 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 8 37 theme PCSK9 1383:1387 arg1 forms 1374:1378 both native and furin-like cleaved forms 1339:1378 both native and furin-like cleaved forms of PCSK9 1339:1387 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 3 38 theme intracellular 462:474 arg1 degradation 476:486 the intracellular degradation 458:486 the intracellular degradation of the low density lipoprotein (LDL) receptor 458:532 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 6 39 theme motif 1002:1006 arg1 arrow 1025:1029 the motif RFHR218 downward arrow 998:1029 the motif RFHR218 downward arrow 998:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 7 40 theme PCSK9 1251:1255 arg1 inability 1238:1246 the inability 1234:1246 the inability of PCSK9 to exit the endoplasmic reticulum (C679X) 1234:1297 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 10 41 theme cell-surface 1613:1624 arg1 receptor 1630:1637 the cell-surface LDL receptor 1609:1637 the cell-surface LDL receptor 1609:1637 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 4 42 theme post-translational 606:623 arg1 modifications 625:637 the post-translational modifications 602:637 the post-translational modifications of PCSK9 602:646 We analyzed the post-translational modifications of PCSK9 and show that it is sulfated within its prosegment at Tyr38. 16912035 4 43 theme PCSK9 642:646 arg1 modifications 625:637 the post-translational modifications 602:637 the post-translational modifications of PCSK9 602:646 We analyzed the post-translational modifications of PCSK9 and show that it is sulfated within its prosegment at Tyr38. 16912035 1 44 theme proprotein 192:201 arg1 family 219:224 the proprotein convertase (PC) family 188:224 the proprotein convertase (PC) family 188:224 PCSK9 is the ninth member of the proprotein convertase (PC) family. 16912035 3 45 theme action 396:401 arg1 mechanism 383:391 The exact mechanism 373:391 The exact mechanism of action of PCSK9 373:410 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 3 45 theme action 396:401 arg1 clear 419:423 clear 419:423 clear 419:423 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 0 46 theme mutations 111:119 arg1 consequences 87:98 functional consequences 76:98 functional consequences of natural mutations and post-translational modifications 76:156 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 8 47 theme circulating 1392:1402 arg1 plasma 1410:1415 circulating human plasma 1392:1415 circulating human plasma 1392:1415 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 1 48 theme convertase 203:212 arg1 family 219:224 the proprotein convertase (PC) family 188:224 the proprotein convertase (PC) family 188:224 PCSK9 is the ninth member of the proprotein convertase (PC) family. 16912035 3 49 theme exact 377:381 arg1 mechanism 383:391 The exact mechanism 373:391 The exact mechanism of action of PCSK9 373:410 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 3 49 theme exact 377:381 arg1 clear 419:423 clear 419:423 clear 419:423 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 0 50 theme modifications 144:156 arg1 consequences 87:98 functional consequences 76:98 functional consequences of natural mutations and post-translational modifications 76:156 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 8 51 theme native 1344:1349 arg1 forms 1374:1378 both native and furin-like cleaved forms 1339:1378 both native and furin-like cleaved forms of PCSK9 1339:1387 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 2 52 theme dominant 316:323 arg1 form 325:328 a dominant form 314:328 a dominant form of familial hyper- or hypocholesterolemia 314:370 Some of its natural mutations have been genetically associated with the development of a dominant form of familial hyper- or hypocholesterolemia. 16912035 5 53 theme proteolytic 757:767 arg1 cleavage 769:776 proteolytic cleavage 757:776 proteolytic cleavage by the other members of the PC family 757:814 We also examined the susceptibility of PCSK9 to proteolytic cleavage by the other members of the PC family. 16912035 0 54 theme post-translational 125:142 arg1 modifications 144:156 post-translational modifications 125:156 post-translational modifications 125:156 The proprotein convertase (PC) PCSK9 is inactivated by furin and/or PC5/6A: functional consequences of natural mutations and post-translational modifications. 16912035 3 55 theme acidic 537:542 arg1 compartments 544:555 acidic compartments 537:555 acidic compartments 537:555 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 9 56 theme acid 1493:1496 arg1 convertases 1498:1508 the basic amino acid convertases 1477:1508 the basic amino acid convertases furin and PC5/6A 1477:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 56 theme acid 1493:1496 arg1 PC5/6A 1520:1525 PC5/6A 1520:1525 PC5/6A 1520:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 56 theme acid 1493:1496 arg1 furin 1510:1514 furin 1510:1514 furin 1510:1514 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 10 57 theme LDL 1685:1687 arg1 cholesterol 1689:1699 circulating LDL cholesterol 1673:1699 circulating LDL cholesterol 1673:1699 The latter may reduce the lifetime of this proteinase and its ability to degrade the cell-surface LDL receptor, thereby regulating the levels of circulating LDL cholesterol. 16912035 8 58 attach presence 1327:1334 arg2 forms 1374:1378 both native and furin-like cleaved forms 1339:1378 both native and furin-like cleaved forms of PCSK9 1339:1387 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 8 58 attach presence 1327:1334 arg1 plasma 1410:1415 circulating human plasma 1392:1415 circulating human plasma 1392:1415 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 8 59 theme human 1404:1408 arg1 plasma 1410:1415 circulating human plasma 1392:1415 circulating human plasma 1392:1415 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 8 60 theme furin-like 1355:1364 arg1 forms 1374:1378 both native and furin-like cleaved forms 1339:1378 both native and furin-like cleaved forms of PCSK9 1339:1387 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 6 61 theme furin/PC5/6A 971:982 arg1 processing 984:993 furin/PC5/6A processing 971:993 furin/PC5/6A processing 971:993 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 6 62 theme processing 984:993 arg1 loss 963:966 total or partial loss 946:966 total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow 946:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 3 63 theme low 495:497 arg1 LDL 520:522 LDL 520:522 LDL 520:522 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 3 63 theme low 495:497 arg1 lipoprotein 507:517 low density lipoprotein 495:517 the low density lipoprotein (LDL) receptor 491:532 The exact mechanism of action of PCSK9 is not clear, although it is known to enhance the intracellular degradation of the low density lipoprotein (LDL) receptor in acidic compartments, likely the endosomes/lysosomes. 16912035 6 64 theme downward 1016:1023 arg1 arrow 1025:1029 the motif RFHR218 downward arrow 998:1029 the motif RFHR218 downward arrow 998:1029 The data show that the natural gain-of-function mutations R218S, F216L, and D374Y associated with hypercholesterolemia result in total or partial loss of furin/PC5/6A processing at the motif RFHR218 downward arrow. 16912035 1 65 theme PC 215:216 arg1 family 219:224 the proprotein convertase (PC) family 188:224 the proprotein convertase (PC) family 188:224 PCSK9 is the ninth member of the proprotein convertase (PC) family. 16912035 9 66 dep convertases 1498:1508 arg1 convertases 1498:1508 the basic amino acid convertases 1477:1508 the basic amino acid convertases furin and PC5/6A 1477:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 66 dep convertases 1498:1508 arg1 PC5/6A 1520:1525 PC5/6A 1520:1525 PC5/6A 1520:1525 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 9 66 dep convertases 1498:1508 arg1 furin 1510:1514 furin 1510:1514 furin 1510:1514 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 5 67 theme PC 806:807 arg1 family 809:814 the PC family 802:814 the PC family 802:814 We also examined the susceptibility of PCSK9 to proteolytic cleavage by the other members of the PC family. 16912035 8 68 theme forms 1374:1378 arg1 presence 1327:1334 the presence 1323:1334 the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma 1323:1415 Furthermore, we report the presence of both native and furin-like cleaved forms of PCSK9 in circulating human plasma. 16912035 9 69 theme PCSK9 1440:1444 arg1 levels 1446:1451 PCSK9 levels 1440:1451 PCSK9 levels 1440:1451 Thus, we propose that PCSK9 levels are finely regulated by the basic amino acid convertases furin and PC5/6A. 16912035 2 70 theme hyper- 342:347 arg1 form 325:328 a dominant form 314:328 a dominant form of familial hyper- or hypocholesterolemia 314:370 Some of its natural mutations have been genetically associated with the development of a dominant form of familial hyper- or hypocholesterolemia. 16912035 7 71 theme furin 1205:1209 arg1 A443T 1221:1225 A443T 1221:1225 A443T 1221:1225 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 7 71 theme furin 1205:1209 arg1 cleavage 1211:1218 furin cleavage 1205:1218 furin cleavage (A443T) 1205:1226 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 5 72 theme other 785:789 arg1 members 791:797 the other members 781:797 the other members of the PC family 781:814 We also examined the susceptibility of PCSK9 to proteolytic cleavage by the other members of the PC family. 16912035 7 73 theme trans-Golgi 1119:1129 arg1 localization 1158:1169 trans-Golgi network/recycling endosome localization 1119:1169 trans-Golgi network/recycling endosome localization 1119:1169 In contrast, the loss-of-function mutations A443T and C679X lead either to the lack of trans-Golgi network/recycling endosome localization and an enhanced susceptibility to furin cleavage (A443T) or to the inability of PCSK9 to exit the endoplasmic reticulum (C679X). 16912035 1 74 theme family 219:224 arg1 member 178:183 the ninth member 168:183 the ninth member of the proprotein convertase (PC) family 168:224 PCSK9 is the ninth member of the proprotein convertase (PC) family. 16912035 1 74 theme family 219:224 arg1 PCSK9 159:163 PCSK9 159:163 PCSK9 159:163 PCSK9 is the ninth member of the proprotein convertase (PC) family. 3402609 2 0 theme long 240:243 arg1 protein 245:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 2 1 theme acid 235:238 arg1 protein 245:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 0 2 theme amino 9:13 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of a human platelet proteoglycan 0:60 Complete amino acid sequence of a human platelet proteoglycan. 3402609 0 3 theme Complete 0:7 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of a human platelet proteoglycan 0:60 Complete amino acid sequence of a human platelet proteoglycan. 3402609 3 4 theme promyelocytic 342:354 arg1 line 370:373 promyelocytic leukemia cell line 342:373 promyelocytic leukemia cell line 342:373 The significance of homologies observed between P.PG and promyelocytic leukemia cell line proteoglycans is discussed. 3402609 2 5 theme Ser-Gly 268:274 arg1 repeats 276:282 eight Ser-Gly repeats 262:282 eight Ser-Gly repeats 262:282 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 1 6 theme amino 166:170 arg1 analysis 186:193 amino acid sequence analysis 166:193 amino acid sequence analysis 166:193 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 0 7 theme acid 15:18 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of a human platelet proteoglycan 0:60 Complete amino acid sequence of a human platelet proteoglycan. 3402609 1 8 theme human 90:94 arg1 core 125:128 a human platelet proteoglycan (P.PG) core 88:128 a human platelet proteoglycan (P.PG) core 88:128 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 1 9 theme acid 172:175 arg1 analysis 186:193 amino acid sequence analysis 166:193 amino acid sequence analysis 166:193 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 2 10 contain contains 253:260 arg1 protein 245:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 2 10 contain contains 253:260 arg2 repeats 276:282 eight Ser-Gly repeats 262:282 eight Ser-Gly repeats 262:282 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 1 11 theme sequence 177:184 arg1 analysis 186:193 amino acid sequence analysis 166:193 amino acid sequence analysis 166:193 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 1 12 theme analysis 186:193 arg1 combination 151:161 a combination 149:161 a combination of amino acid sequence analysis and cDNA cloning 149:210 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 0 13 theme platelet 40:47 arg1 proteoglycan 49:60 a human platelet proteoglycan 32:60 a human platelet proteoglycan 32:60 Complete amino acid sequence of a human platelet proteoglycan. 3402609 3 14 theme homologies 305:314 arg1 significance 289:300 The significance 285:300 The significance of homologies observed between P.PG and promyelocytic leukemia cell line proteoglycans 285:387 The significance of homologies observed between P.PG and promyelocytic leukemia cell line proteoglycans is discussed. 3402609 1 15 theme platelet 96:103 arg1 core 125:128 a human platelet proteoglycan (P.PG) core 88:128 a human platelet proteoglycan (P.PG) core 88:128 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 0 16 theme human 34:38 arg1 proteoglycan 49:60 a human platelet proteoglycan 32:60 a human platelet proteoglycan 32:60 Complete amino acid sequence of a human platelet proteoglycan. 3402609 1 17 theme proteoglycan 105:116 arg1 core 125:128 a human platelet proteoglycan (P.PG) core 88:128 a human platelet proteoglycan (P.PG) core 88:128 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 3 18 dep P.PG 333:336 arg1 proteoglycans 375:387 proteoglycans 375:387 proteoglycans 375:387 The significance of homologies observed between P.PG and promyelocytic leukemia cell line proteoglycans is discussed. 3402609 2 19 theme amino 229:233 arg1 protein 245:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 1 20 theme primary 67:73 arg1 structure 75:83 The primary structure 63:83 The primary structure of a human platelet proteoglycan (P.PG) core 63:128 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 2 21 theme 131 225:227 arg1 protein 245:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 1 22 theme P.PG 119:122 arg1 core 125:128 a human platelet proteoglycan (P.PG) core 88:128 a human platelet proteoglycan (P.PG) core 88:128 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 1 23 theme cDNA 199:202 arg1 cloning 204:210 cDNA cloning 199:210 cDNA cloning 199:210 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 2 24 theme deduced 217:223 arg1 protein 245:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein 213:251 The deduced 131 amino acid long protein contains eight Ser-Gly repeats. 3402609 3 25 theme cell 365:368 arg1 line 370:373 promyelocytic leukemia cell line 342:373 promyelocytic leukemia cell line 342:373 The significance of homologies observed between P.PG and promyelocytic leukemia cell line proteoglycans is discussed. 3402609 1 26 theme cloning 204:210 arg1 combination 151:161 a combination 149:161 a combination of amino acid sequence analysis and cDNA cloning 149:210 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 3402609 0 27 theme proteoglycan 49:60 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of a human platelet proteoglycan 0:60 Complete amino acid sequence of a human platelet proteoglycan. 3402609 3 28 theme leukemia 356:363 arg1 line 370:373 promyelocytic leukemia cell line 342:373 promyelocytic leukemia cell line 342:373 The significance of homologies observed between P.PG and promyelocytic leukemia cell line proteoglycans is discussed. 3402609 1 29 theme core 125:128 arg1 structure 75:83 The primary structure 63:83 The primary structure of a human platelet proteoglycan (P.PG) core 63:128 The primary structure of a human platelet proteoglycan (P.PG) core was established by a combination of amino acid sequence analysis and cDNA cloning. 17542669 8 0 theme autoantibody 1205:1216 arg1 arrangements 1185:1196 The receptor-binding arrangements 1164:1196 The receptor-binding arrangements of the autoantibody 1164:1216 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 8 0 theme autoantibody 1205:1216 arg1 similar 1227:1233 similar 1227:1233 similar 1227:1233 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 10 1 with structure 1602:1610 arg1 M22 1645:1647 M22 1645:1647 M22 1645:1647 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 6 2 theme lock 923:926 arg1 " 935:935 "lock and key" 922:935 "lock and key" binding 922:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 1 3 theme molecular 249:257 arg1 level 259:263 the molecular level 245:263 the molecular level 245:263 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 0 4 theme thyroid-stimulating 56:74 arg1 autoantibody 76:87 a thyroid-stimulating autoantibody 54:87 a thyroid-stimulating autoantibody 54:87 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 17542669 0 5 from structure 8:16 arg1 complex 41:47 complex 41:47 complex with a thyroid-stimulating autoantibody 41:87 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 17542669 7 6 theme M22 1121:1123 arg1 interactions 1105:1116 interactions 1105:1116 interactions of M22 with intact, functionally active TSHR 1105:1161 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 4 7 theme curved 543:548 arg1 tube 558:561 a curved helical tube 541:561 a curved helical tube 541:561 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 7 8 theme residues 958:965 arg1 Mutation 946:953 Mutation 946:953 Mutation of residues showing strong interactions in the structure 946:1010 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 7 9 theme strong 975:980 arg1 interactions 982:993 strong interactions 975:993 strong interactions 975:993 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 10 10 theme new 1684:1686 arg1 strategies 1688:1697 new strategies 1684:1697 new strategies 1684:1697 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 2 11 theme part 286:289 arg1 complex 275:281 A complex 273:281 A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab 273:368 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 8 12 theme follicle-stimulating 1257:1276 arg1 FSH 1287:1289 FSH 1287:1289 FSH 1287:1289 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 8 12 theme follicle-stimulating 1257:1276 arg1 hormone 1278:1284 follicle-stimulating hormone 1257:1284 follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself 1257:1378 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 6 13 theme binding 864:870 arg1 interface 872:880 the binding interface 860:880 the binding interface compared to unbound M22 consistent with "lock and key" binding 860:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 9 14 theme identical 1465:1473 arg1 features 1492:1499 almost identical receptor-binding features 1458:1499 almost identical receptor-binding features 1458:1499 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 7 15 theme M22 1023:1025 arg1 activity 1027:1034 M22 activity 1023:1034 M22 activity 1023:1034 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 2 16 theme domain 317:322 arg1 part 286:289 part 286:289 part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab 286:368 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 7 17 theme binding 1057:1063 arg1 detail 1065:1070 the binding detail 1053:1070 the binding detail observed in the complex 1053:1094 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 5 18 theme polar 734:738 arg1 bonding 757:763 ionic, polar, and hydrophobic bonding 727:763 ionic, polar, and hydrophobic bonding 727:763 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 4 19 theme concave 586:592 arg1 surface 594:600 its concave surface 582:600 its concave surface 582:600 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 4 20 theme tube 623:626 arg1 axis 635:638 the tube length axis 619:638 the tube length axis 619:638 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 3 21 theme suitable 406:413 arg1 Crystals 397:404 Crystals 397:404 Crystals suitable for X-ray diffraction analysis 397:444 Crystals suitable for X-ray diffraction analysis were obtained and the structure solved at 2.55 A resolution. 17542669 4 22 theme helical 550:556 arg1 tube 558:561 a curved helical tube 541:561 a curved helical tube 541:561 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 9 23 theme thyroid-stimulating 1419:1437 arg1 autoantibody 1439:1450 the thyroid-stimulating autoantibody 1415:1450 the thyroid-stimulating autoantibody 1415:1450 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 2 24 theme M22 362:364 arg1 Fab 366:368 M22 Fab 362:368 M22 Fab 362:368 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 5 25 theme extensive 706:714 arg1 network 716:722 an extensive network 703:722 an extensive network of ionic, polar, and hydrophobic bonding 703:763 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 7 26 theme intact 1130:1135 arg1 TSHR 1158:1161 intact, functionally active TSHR 1130:1161 intact, functionally active TSHR 1130:1161 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 2 27 theme extracellular 303:315 arg1 domain 317:322 the TSHR extracellular domain 294:322 the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab 294:368 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 2 27 theme extracellular 303:315 arg1 acids 331:335 amino acids 1-260; TSHR260 325:350 amino acids 1-260; TSHR260 325:350 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 6 28 theme " 935:935 arg1 binding 937:943 "lock and key" binding 922:943 "lock and key" binding 922:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 2 29 dep DESIGN 266:271 arg1 prepared 374:381 prepared 374:381 prepared 374:381 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 2 29 dep DESIGN 266:271 arg1 purified 387:394 purified 387:394 purified 387:394 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 0 30 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 0:88 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 17542669 7 31 from Mutation 946:953 arg1 structure 1002:1010 the structure 998:1010 the structure 998:1010 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 5 32 theme hydrophobic 745:755 arg1 bonding 757:763 ionic, polar, and hydrophobic bonding 727:763 ionic, polar, and hydrophobic bonding 727:763 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 2 33 theme TSHR 298:301 arg1 domain 317:322 the TSHR extracellular domain 294:322 the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab 294:368 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 2 33 theme TSHR 298:301 arg1 acids 331:335 amino acids 1-260; TSHR260 325:350 amino acids 1-260; TSHR260 325:350 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 1 34 theme thyroid-stimulating 186:204 arg1 M22 237:239 M22 237:239 M22 237:239 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 1 34 theme thyroid-stimulating 186:204 arg1 autoantibody 223:234 a thyroid-stimulating human monoclonal autoantibody 184:234 a thyroid-stimulating human monoclonal autoantibody (M22) 184:240 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 6 35 theme residues 848:855 arg1 atoms 835:839 the atoms 831:839 the atoms of M22 residues 831:855 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 8 36 theme receptor-binding 1168:1183 arg1 arrangements 1185:1196 The receptor-binding arrangements 1164:1196 The receptor-binding arrangements of the autoantibody 1164:1216 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 8 36 theme receptor-binding 1168:1183 arg1 similar 1227:1233 similar 1227:1233 similar 1227:1233 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 2 37 dep acids 331:335 arg1 TSHR260 344:350 TSHR260 344:350 TSHR260 344:350 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 1 38 theme human 206:210 arg1 M22 237:239 M22 237:239 M22 237:239 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 1 38 theme human 206:210 arg1 autoantibody 223:234 a thyroid-stimulating human monoclonal autoantibody 184:234 a thyroid-stimulating human monoclonal autoantibody (M22) 184:240 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 5 39 theme ionic 727:731 arg1 bonding 757:763 ionic, polar, and hydrophobic bonding 727:763 ionic, polar, and hydrophobic bonding 727:763 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 6 40 theme M22 844:846 arg1 residues 848:855 M22 residues 844:855 M22 residues 844:855 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 10 41 theme TSHR 1619:1622 arg1 complex 1632:1638 its complex 1628:1638 its complex with M22 1628:1647 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 10 41 theme TSHR 1619:1622 arg1 structure 1602:1610 our structure 1598:1610 our structure of the TSHR 1598:1622 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 8 42 theme amino 1321:1325 arg1 acids 1327:1331 amino acids 1-268 1321:1337 amino acids 1-268 1321:1337 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 8 42 theme amino 1321:1325 arg1 receptor 1311:1318 the FSH receptor 1303:1318 the FSH receptor (amino acids 1-268) 1303:1338 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 7 43 dep intact 1130:1135 arg1 active 1151:1156 active 1151:1156 active 1151:1156 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 1 44 theme monoclonal 212:221 arg1 M22 237:239 M22 237:239 M22 237:239 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 1 44 theme monoclonal 212:221 arg1 autoantibody 223:234 a thyroid-stimulating human monoclonal autoantibody 184:234 a thyroid-stimulating human monoclonal autoantibody (M22) 184:240 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 0 45 theme receptor 29:36 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 0:88 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 17542669 10 46 gly glycoprotein 1730:1741 arg1 glycoprotein 1730:1741 glycoprotein hormone receptor activation 1730:1769 glycoprotein hormone receptor activation 1730:1769 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 9 47 theme receptor-binding 1475:1490 arg1 features 1492:1499 almost identical receptor-binding features 1458:1499 almost identical receptor-binding features 1458:1499 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 6 48 theme key 932:934 arg1 " 935:935 "lock and key" 922:935 "lock and key" binding 922:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 3 49 theme X-ray 419:423 arg1 analysis 437:444 X-ray diffraction analysis 419:444 X-ray diffraction analysis 419:444 Crystals suitable for X-ray diffraction analysis were obtained and the structure solved at 2.55 A resolution. 17542669 0 50 theme TSH 25:27 arg1 receptor 29:36 the TSH receptor 21:36 the TSH receptor 21:36 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 17542669 10 51 theme hormone 1743:1749 arg1 activation 1760:1769 glycoprotein hormone receptor activation 1730:1769 glycoprotein hormone receptor activation 1730:1769 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 3 52 theme diffraction 425:435 arg1 analysis 437:444 X-ray diffraction analysis 419:444 X-ray diffraction analysis 419:444 Crystals suitable for X-ray diffraction analysis were obtained and the structure solved at 2.55 A resolution. 17542669 5 53 dep large 680:684 arg1 large 680:684 large 680:684 large 680:684 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 5 53 dep large 680:684 arg1 A 693:693 2,500 A 687:693 2,500 A(2) 687:696 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 5 53 dep large 680:684 arg1 interface 645:653 The interface 641:653 The interface buried in the complex 641:675 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 5 53 dep large 680:684 arg1 2 695:695 2 695:695 2 695:695 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 10 54 theme glycoprotein 1730:1741 arg1 activation 1760:1769 glycoprotein hormone receptor activation 1730:1769 glycoprotein hormone receptor activation 1730:1769 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 6 55 with consistent 906:915 arg1 binding 937:943 "lock and key" binding 922:943 "lock and key" binding 922:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 7 56 located observed 1072:1079 arg1 complex 1088:1094 the complex 1084:1094 the complex 1084:1094 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 7 56 located observed 1072:1079 arg2 detail 1065:1070 the binding detail 1053:1070 the binding detail observed in the complex 1053:1094 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 10 57 theme receptor 1751:1758 arg1 activation 1760:1769 glycoprotein hormone receptor activation 1730:1769 glycoprotein hormone receptor activation 1730:1769 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 6 58 from movement 819:826 arg1 interface 872:880 the binding interface 860:880 the binding interface compared to unbound M22 consistent with "lock and key" binding 860:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 6 58 from movement 819:826 arg1 atoms 835:839 the atoms 831:839 the atoms of M22 residues 831:855 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 8 59 theme FSH 1307:1309 arg1 acids 1327:1331 amino acids 1-268 1321:1337 amino acids 1-268 1321:1337 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 8 59 theme FSH 1307:1309 arg1 receptor 1311:1318 the FSH receptor 1303:1318 the FSH receptor (amino acids 1-268) 1303:1338 The receptor-binding arrangements of the autoantibody are very similar to those reported for follicle-stimulating hormone (FSH) binding to the FSH receptor (amino acids 1-268) and consequently to those of TSH itself. 17542669 2 60 theme amino 325:329 arg1 domain 317:322 the TSHR extracellular domain 294:322 the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab 294:368 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 2 60 theme amino 325:329 arg1 acids 331:335 amino acids 1-260; TSHR260 325:350 amino acids 1-260; TSHR260 325:350 DESIGN A complex of part of the TSHR extracellular domain (amino acids 1-260; TSHR260) bound to M22 Fab was prepared and purified. 17542669 4 61 theme MAIN 507:510 arg1 OUTCOME 512:518 MAIN OUTCOME TSHR260 507:526 MAIN OUTCOME TSHR260 507:526 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 10 62 theme autoimmune 1779:1788 arg1 response 1790:1797 the autoimmune response 1775:1797 the autoimmune response to the TSHR 1775:1809 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 17542669 4 63 theme length 628:633 arg1 axis 635:638 the tube length axis 619:638 the tube length axis 619:638 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 6 64 theme consistent 906:915 arg1 M22 902:904 unbound M22 894:904 unbound M22 consistent with "lock and key" binding 894:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 0 65 with complex 41:47 arg1 autoantibody 76:87 a thyroid-stimulating autoantibody 54:87 a thyroid-stimulating autoantibody 54:87 Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. 17542669 1 66 theme thyroid-stimulating 136:154 arg1 receptor 164:171 the thyroid-stimulating hormone receptor 132:171 the thyroid-stimulating hormone receptor (TSHR) 132:178 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 1 66 theme thyroid-stimulating 136:154 arg1 TSHR 174:177 TSHR 174:177 TSHR 174:177 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 4 67 theme M22 567:569 arg1 Fab 571:573 M22 Fab 567:573 M22 Fab 567:573 MAIN OUTCOME TSHR260 comprises of a curved helical tube and M22 Fab clasps its concave surface at 90 degrees to the tube length axis. 17542669 5 68 theme 2,500 687:691 arg1 large 680:684 large 680:684 large 680:684 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 5 68 theme 2,500 687:691 arg1 A 693:693 2,500 A 687:693 2,500 A(2) 687:696 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 5 68 theme 2,500 687:691 arg1 interface 645:653 The interface 641:653 The interface buried in the complex 641:675 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 5 68 theme 2,500 687:691 arg1 2 695:695 2 695:695 2 695:695 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 9 69 dep structures 1521:1530 arg1 the 1517:1519 the 1517:1519 the 1517:1519 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 1 70 theme hormone 156:162 arg1 receptor 164:171 the thyroid-stimulating hormone receptor 132:171 the thyroid-stimulating hormone receptor (TSHR) 132:178 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 1 70 theme hormone 156:162 arg1 TSHR 174:177 TSHR 174:177 TSHR 174:177 OBJECTIVE To analyze interactions between the thyroid-stimulating hormone receptor (TSHR) and a thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. 17542669 5 71 theme bonding 757:763 arg1 network 716:722 an extensive network 703:722 an extensive network of ionic, polar, and hydrophobic bonding 703:763 The interface buried in the complex is large (2,500 A(2)) and an extensive network of ionic, polar, and hydrophobic bonding is involved in the interaction. 17542669 6 72 theme unbound 894:900 arg1 M22 902:904 unbound M22 894:904 unbound M22 consistent with "lock and key" binding 894:943 There is virtually no movement in the atoms of M22 residues on the binding interface compared to unbound M22 consistent with "lock and key" binding. 17542669 9 73 theme ligands 1557:1563 arg1 origins 1536:1542 origins 1536:1542 origins 1536:1542 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 9 73 theme ligands 1557:1563 arg1 structures 1521:1530 structures 1521:1530 structures 1521:1530 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 7 74 with interactions 1105:1116 arg1 TSHR 1158:1161 intact, functionally active TSHR 1130:1161 intact, functionally active TSHR 1130:1161 Mutation of residues showing strong interactions in the structure influenced M22 activity, indicating that the binding detail observed in the complex reflects interactions of M22 with intact, functionally active TSHR. 17542669 3 75 theme A 493:493 arg1 resolution 495:504 2.55 A resolution 488:504 2.55 A resolution 488:504 Crystals suitable for X-ray diffraction analysis were obtained and the structure solved at 2.55 A resolution. 17542669 9 76 dep CONCLUSIONS 1381:1391 arg1 remarkable 1399:1408 remarkable 1399:1408 remarkable 1399:1408 CONCLUSIONS It is remarkable that the thyroid-stimulating autoantibody shows almost identical receptor-binding features to TSH although the structures and origins of these two ligands are very different. 17542669 10 77 with complex 1632:1638 arg1 M22 1645:1647 M22 1645:1647 M22 1645:1647 Furthermore, our structure of the TSHR and its complex with M22 provide foundations for developing new strategies to understand and control both glycoprotein hormone receptor activation and the autoimmune response to the TSHR. 7682553 0 0 theme heavy 128:132 arg1 2/bikunin 140:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 7 1 theme mass 959:962 arg1 analysis 978:985 mass spectrometric analysis 959:985 mass spectrometric analysis 959:985 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 2 2 theme polypeptide 436:446 arg1 chains 448:453 these two polypeptide chains 426:453 these two polypeptide chains 426:453 This observation suggests that a nondisulfide cross-link is responsible for the association of these two polypeptide chains. 7682553 4 3 theme chondroitin 645:655 arg1 enzymes 675:681 chondroitin sulfate-degrading enzymes 645:681 chondroitin sulfate-degrading enzymes 645:681 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 0 4 theme inhibitor 118:126 arg1 2/bikunin 140:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 7 5 theme typical 1114:1120 arg1 link 1135:1138 a typical O-glycosidic link 1112:1138 a typical O-glycosidic link to Ser10 of bikunin 1112:1158 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 0 6 from Presence 0:7 arg1 2/bikunin 140:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 2 7 theme nondisulfide 364:375 arg1 responsible 391:401 responsible 391:401 responsible 391:401 This observation suggests that a nondisulfide cross-link is responsible for the association of these two polypeptide chains. 7682553 2 7 theme nondisulfide 364:375 arg1 cross-link 377:386 a nondisulfide cross-link 362:386 a nondisulfide cross-link 362:386 This observation suggests that a nondisulfide cross-link is responsible for the association of these two polypeptide chains. 7682553 0 8 theme chain 134:138 arg1 2/bikunin 140:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 4 9 located found 772:776 arg2 B. 869:870 B. 869:870 B. 869:870 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 cross-link 761:770 the protein-glycosaminoglycan-protein cross-link 723:770 the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol 723:919 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 J. 823:824 J. 823:824 J. 823:824 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 Salvesen 830:837 Salvesen 830:837 Salvesen 830:837 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 S. 851:852 S. 851:852 S. 851:852 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 S. 900:901 S. 900:901 S. 900:901 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 G. 840:841 G. 840:841 G. 840:841 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 Thøgersen 855:863 Thøgersen 855:863 Thøgersen 855:863 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 Hefta 844:848 Hefta 844:848 Hefta 844:848 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 I. 866:867 I. 866:867 I. 866:867 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg1 pre-alpha-inhibitor 793:811 the related pre-alpha-inhibitor 781:811 the related pre-alpha-inhibitor 781:811 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 9 located found 772:776 arg2 S. 885:886 S. 885:886 S. 885:886 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 10 theme related 785:791 arg1 pre-alpha-inhibitor 793:811 the related pre-alpha-inhibitor 781:811 the related pre-alpha-inhibitor 781:811 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 11 theme sulfate-degrading 657:673 arg1 enzymes 675:681 chondroitin sulfate-degrading enzymes 645:681 chondroitin sulfate-degrading enzymes 645:681 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 7 12 contain containing 1003:1012 arg1 peptides 994:1001 the peptides 990:1001 the peptides containing the cross-link 990:1027 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 7 12 contain containing 1003:1012 arg2 cross-link 1018:1027 the cross-link 1014:1027 the cross-link 1014:1027 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 8 13 theme N-acetylgalactosamine 1268:1288 arg1 C-6 1249:1251 C-6 1249:1251 C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain 1249:1323 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 3 14 theme techniques 501:510 arg1 techniques 501:510 techniques 501:510 techniques 501:510 In this study, we have utilized a variety of techniques to investigate the structural basis for this observation. 7682553 3 14 theme techniques 501:510 arg1 variety 490:496 a variety 488:496 a variety of techniques 488:510 In this study, we have utilized a variety of techniques to investigate the structural basis for this observation. 7682553 8 15 theme chain 1203:1207 arg1 residue 1186:1192 The COOH-terminal Asp648 residue 1161:1192 The COOH-terminal Asp648 residue of heavy chain 2 1161:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 8 15 theme chain 1203:1207 arg1 chain 1203:1207 heavy chain 2 1197:1209 heavy chain 2 1197:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 4 16 dep found 772:776 arg1 Thøgersen 855:863 Thøgersen 855:863 Thøgersen 855:863 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 S. 885:886 S. 885:886 S. 885:886 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 G. 840:841 G. 840:841 G. 840:841 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 Hefta 844:848 Hefta 844:848 Hefta 844:848 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 cross-link 761:770 the protein-glycosaminoglycan-protein cross-link 723:770 the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol 723:919 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 I. 866:867 I. 866:867 I. 866:867 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 J. 823:824 J. 823:824 J. 823:824 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 1991 907:910 1991 907:910 1991 907:910 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 B. 869:870 B. 869:870 B. 869:870 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 Salvesen 830:837 Salvesen 830:837 Salvesen 830:837 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 S. 851:852 S. 851:852 S. 851:852 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 16 dep found 772:776 arg1 S. 900:901 S. 900:901 S. 900:901 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 1 17 theme polypeptide 218:228 arg1 chains 230:235 two polypeptide chains 214:235 two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis 214:328 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 8 18 theme Asp648 1179:1184 arg1 residue 1186:1192 The COOH-terminal Asp648 residue 1161:1192 The COOH-terminal Asp648 residue of heavy chain 2 1161:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 8 18 theme Asp648 1179:1184 arg1 chain 1203:1207 heavy chain 2 1197:1209 heavy chain 2 1197:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 8 19 theme COOH-terminal 1165:1177 arg1 residue 1186:1192 The COOH-terminal Asp648 residue 1161:1192 The COOH-terminal Asp648 residue of heavy chain 2 1161:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 8 19 theme COOH-terminal 1165:1177 arg1 chain 1203:1207 heavy chain 2 1197:1209 heavy chain 2 1197:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 7 20 theme chondroitin-4-sulfate 1063:1083 arg1 chain 1085:1089 a chondroitin-4-sulfate chain 1061:1089 a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin 1061:1158 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 Thøgersen 855:863 Thøgersen 855:863 Thøgersen 855:863 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 S. 885:886 S. 885:886 S. 885:886 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 G. 840:841 G. 840:841 G. 840:841 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 S. 900:901 S. 900:901 S. 900:901 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 Hefta 844:848 Hefta 844:848 Hefta 844:848 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 I. 866:867 I. 866:867 I. 866:867 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 J. 823:824 J. 823:824 J. 823:824 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 B. 869:870 B. 869:870 B. 869:870 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 Salvesen 830:837 Salvesen 830:837 Salvesen 830:837 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 S. 851:852 S. 851:852 S. 851:852 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 21 theme protein-glycosaminoglycan-protein 727:759 arg1 cross-link 761:770 the protein-glycosaminoglycan-protein cross-link 723:770 the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol 723:919 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 0 22 theme protein-glycosaminoglycan-protein 16:48 arg1 cross-link 59:68 the protein-glycosaminoglycan-protein covalent cross-link 12:68 the protein-glycosaminoglycan-protein covalent cross-link 12:68 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 4 23 theme protein 614:620 arg1 chains 622:627 the two protein chains 606:627 the two protein chains 606:627 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 3 24 theme structural 531:540 arg1 basis 542:546 the structural basis 527:546 the structural basis for this observation 527:567 In this study, we have utilized a variety of techniques to investigate the structural basis for this observation. 7682553 7 25 theme bikunin 1152:1158 arg1 Ser10 1143:1147 Ser10 1143:1147 Ser10 of bikunin 1143:1158 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 4 26 dep B. 869:870 arg1 Biol 916:919 Biol 916:919 Biol 916:919 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 0 27 theme cross-link 59:68 arg1 Presence 0:7 Presence 0:7 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 0:149 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 9 28 theme protein-glycosaminoglycan-protein 1349:1381 arg1 identical 1446:1454 identical 1446:1454 identical 1446:1454 This suggests that the protein-glycosaminoglycan-protein cross-link that assembles the chains of pre-alpha-inhibitor is identical to that which assembles HC2/bikunin, and is probably a characteristic of the bikunin proteins. 7682553 9 28 theme protein-glycosaminoglycan-protein 1349:1381 arg1 cross-link 1383:1392 the protein-glycosaminoglycan-protein cross-link 1345:1392 the protein-glycosaminoglycan-protein cross-link that assembles the chains of pre-alpha-inhibitor 1345:1441 This suggests that the protein-glycosaminoglycan-protein cross-link that assembles the chains of pre-alpha-inhibitor is identical to that which assembles HC2/bikunin, and is probably a characteristic of the bikunin proteins. 7682553 9 29 theme pre-alpha-inhibitor 1423:1441 arg1 chains 1413:1418 the chains 1409:1418 the chains of pre-alpha-inhibitor 1409:1441 This suggests that the protein-glycosaminoglycan-protein cross-link that assembles the chains of pre-alpha-inhibitor is identical to that which assembles HC2/bikunin, and is probably a characteristic of the bikunin proteins. 7682553 0 30 theme covalent 50:57 arg1 cross-link 59:68 the protein-glycosaminoglycan-protein covalent cross-link 12:68 the protein-glycosaminoglycan-protein covalent cross-link 12:68 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 8 31 theme heavy 1197:1201 arg1 chain 1203:1207 heavy chain 2 1197:1209 heavy chain 2 1197:1209 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 4 32 theme mM 693:694 arg1 properties 702:711 properties 702:711 properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol 702:919 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 4 32 theme mM 693:694 arg1 NaOH 696:699 50 mM NaOH 690:699 50 mM NaOH 690:699 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 7 33 theme peptides 994:1001 arg1 Biochemical 943:953 Biochemical 943:953 Biochemical 943:953 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 1 34 theme reducing 268:275 arg1 conditions 277:286 reducing conditions 268:286 reducing conditions 268:286 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 7 35 theme O-glycosidic 1122:1133 arg1 link 1135:1138 a typical O-glycosidic link 1112:1138 a typical O-glycosidic link to Ser10 of bikunin 1112:1158 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 6 36 dep 266 928:930 arg1 747-751 933:939 747-751 933:939 747-751 933:939 266, 747-751). 7682553 8 37 theme chondroitin-4-sulfate 1297:1317 arg1 chain 1319:1323 the chondroitin-4-sulfate chain 1293:1323 the chondroitin-4-sulfate chain 1293:1323 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 9 38 theme proteins 1541:1548 arg1 characteristic 1511:1524 a characteristic 1509:1524 a characteristic of the bikunin proteins 1509:1548 This suggests that the protein-glycosaminoglycan-protein cross-link that assembles the chains of pre-alpha-inhibitor is identical to that which assembles HC2/bikunin, and is probably a characteristic of the bikunin proteins. 7682553 8 39 theme internal 1259:1266 arg1 N-acetylgalactosamine 1268:1288 an internal N-acetylgalactosamine 1256:1288 an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain 1256:1323 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 1 40 theme human 168:172 arg1 inhibitor 192:200 a human plasma proteinase inhibitor 166:200 a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis 166:328 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 1 40 theme human 168:172 arg1 HC2/bikunin 151:161 HC2/bikunin 151:161 HC2/bikunin 151:161 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 3 41 used utilized 479:486 arg2 we 471:472 we 471:472 we 471:472 In this study, we have utilized a variety of techniques to investigate the structural basis for this observation. 7682553 8 42 theme chain 1319:1323 arg1 N-acetylgalactosamine 1268:1288 an internal N-acetylgalactosamine 1256:1288 an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain 1256:1323 The COOH-terminal Asp648 residue of heavy chain 2 is esterified via the alpha-carbon to C-6 of an internal N-acetylgalactosamine of the chondroitin-4-sulfate chain. 7682553 7 43 theme spectrometric 964:976 arg1 analysis 978:985 mass spectrometric analysis 959:985 mass spectrometric analysis 959:985 Biochemical and mass spectrometric analysis of the peptides containing the cross-link indicate that it is mediated by a chondroitin-4-sulfate chain that originates from a typical O-glycosidic link to Ser10 of bikunin. 7682553 1 44 theme plasma 174:179 arg1 inhibitor 192:200 a human plasma proteinase inhibitor 166:200 a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis 166:328 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 1 44 theme plasma 174:179 arg1 HC2/bikunin 151:161 HC2/bikunin 151:161 HC2/bikunin 151:161 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 9 45 theme bikunin 1533:1539 arg1 proteins 1541:1548 the bikunin proteins 1529:1548 the bikunin proteins 1529:1548 This suggests that the protein-glycosaminoglycan-protein cross-link that assembles the chains of pre-alpha-inhibitor is identical to that which assembles HC2/bikunin, and is probably a characteristic of the bikunin proteins. 7682553 0 46 theme proteinase 107:116 arg1 2/bikunin 140:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 1 47 theme proteinase 181:190 arg1 inhibitor 192:200 a human plasma proteinase inhibitor 166:200 a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis 166:328 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 1 47 theme proteinase 181:190 arg1 HC2/bikunin 151:161 HC2/bikunin 151:161 HC2/bikunin 151:161 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 1 48 theme SDS-polyacrylamide 291:308 arg1 electrophoresis 314:328 SDS-polyacrylamide gel electrophoresis 291:328 SDS-polyacrylamide gel electrophoresis 291:328 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 7682553 0 49 theme inter-alpha-inhibitor-related 77:105 arg1 2/bikunin 140:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin 73:148 Presence of the protein-glycosaminoglycan-protein covalent cross-link in the inter-alpha-inhibitor-related proteinase inhibitor heavy chain 2/bikunin. 7682553 4 50 theme 50 690:691 arg1 mM 693:694 mM 693:694 mM 693:694 We show that the cross-link between the two protein chains is sensitive to chondroitin sulfate-degrading enzymes and to 50 mM NaOH, properties shared by the protein-glycosaminoglycan-protein cross-link found in the related pre-alpha-inhibitor (Enghild, J. J., Salvesen, G., Hefta, S., Thøgersen, I. B., Rutherfurd, S., and Pizzo, S. V. (1991) J. Biol. 7682553 2 51 theme chains 448:453 arg1 association 411:421 the association 407:421 the association of these two polypeptide chains 407:453 This observation suggests that a nondisulfide cross-link is responsible for the association of these two polypeptide chains. 7682553 1 52 theme gel 310:312 arg1 electrophoresis 314:328 SDS-polyacrylamide gel electrophoresis 291:328 SDS-polyacrylamide gel electrophoresis 291:328 HC2/bikunin is a human plasma proteinase inhibitor composed of two polypeptide chains that resist dissociation under reducing conditions in SDS-polyacrylamide gel electrophoresis. 20727575 6 0 theme Plexin-binding 878:891 arg1 determinants 893:904 key Plexin-binding determinants 874:904 key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1 874:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 1 1 theme cardiovascular 236:249 arg1 systems 251:257 cardiovascular systems 236:257 cardiovascular systems 236:257 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 5 2 theme 4c-4d 753:757 arg1 loop 759:762 the Semaphorin's 4c-4d loop 736:762 the Semaphorin's 4c-4d loop 736:762 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 6 3 theme key 874:876 arg1 determinants 893:904 key Plexin-binding determinants 874:904 key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1 874:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 3 4 theme PlexinC1 479:486 arg1 module 469:474 the Semaphorin-binding module 446:474 the Semaphorin-binding module of PlexinC1 446:486 We report the structures of Sema7A and A39R complexed with the Semaphorin-binding module of PlexinC1. 20727575 1 5 theme systems 251:257 arg1 development 180:190 development 180:190 development 180:190 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 1 5 theme systems 251:257 arg1 homeostasis 196:206 homeostasis 196:206 homeostasis 196:206 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 3 6 theme Semaphorin-binding 450:467 arg1 module 469:474 the Semaphorin-binding module 446:474 the Semaphorin-binding module of PlexinC1 446:486 We report the structures of Sema7A and A39R complexed with the Semaphorin-binding module of PlexinC1. 20727575 7 7 theme recognition 1087:1097 arg1 mode 1099:1102 a conserved Semaphorin-Plexin recognition mode 1057:1102 a conserved Semaphorin-Plexin recognition mode 1057:1102 The complex structures support a conserved Semaphorin-Plexin recognition mode and suggest that Plexins are activated by dimerization. 20727575 5 8 theme deep 771:774 arg1 groove 776:781 a deep groove 769:781 a deep groove in blade 3 of the PlexinC1 propeller 769:818 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 5 9 from groove 776:781 arg1 blade 786:790 blade 3 786:792 blade 3 of the PlexinC1 propeller 786:818 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 6 10 theme higher 972:977 arg1 affinity 979:986 higher affinity 972:986 higher affinity binding to the host-derived PlexinC1 972:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 5 11 dep interfaces 691:700 arg1 Both 678:681 Both 678:681 Both 678:681 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 7 12 theme conserved 1059:1067 arg1 mode 1099:1102 a conserved Semaphorin-Plexin recognition mode 1057:1102 a conserved Semaphorin-Plexin recognition mode 1057:1102 The complex structures support a conserved Semaphorin-Plexin recognition mode and suggest that Plexins are activated by dimerization. 20727575 2 13 theme neural 296:301 arg1 Semaphorin 303:312 a neural Semaphorin 294:312 a neural Semaphorin 294:312 Sema7A acts as both an immune and a neural Semaphorin through PlexinC1, and A39R is a Sema7A mimic secreted by smallpox virus. 20727575 2 13 theme neural 296:301 arg1 Sema7A 260:265 Sema7A 260:265 Sema7A 260:265 Sema7A acts as both an immune and a neural Semaphorin through PlexinC1, and A39R is a Sema7A mimic secreted by smallpox virus. 20727575 6 14 link host-derived 1003:1014 arg1 PlexinC1 1016:1023 the host-derived PlexinC1 999:1023 the host-derived PlexinC1 999:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 0 15 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of semaphorin-plexin recognition 0:48 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 7 16 theme Semaphorin-Plexin 1069:1085 arg1 mode 1099:1102 a conserved Semaphorin-Plexin recognition mode 1057:1102 a conserved Semaphorin-Plexin recognition mode 1057:1102 The complex structures support a conserved Semaphorin-Plexin recognition mode and suggest that Plexins are activated by dimerization. 20727575 4 17 theme edge-on 645:651 arg1 orientation 665:675 an edge-on, orthogonal orientation 642:675 orientation 665:675 Both structures show two PlexinC1 molecules symmetrically bridged by Semaphorin dimers, in which the Semaphorin and PlexinC1 beta propellers interact in an edge-on, orthogonal orientation. 20727575 0 18 theme semaphorin-plexin 20:36 arg1 recognition 38:48 semaphorin-plexin recognition 20:48 semaphorin-plexin recognition 20:48 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 5 19 theme binding 683:689 arg1 interfaces 691:700 Both binding interfaces 678:700 interfaces 691:700 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 0 20 with complexes 89:97 arg1 PlexinC1 104:111 PlexinC1 104:111 PlexinC1 104:111 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 3 21 theme A39R 426:429 arg1 structures 401:410 the structures 397:410 the structures of Sema7A and A39R complexed with the Semaphorin-binding module of PlexinC1 397:486 We report the structures of Sema7A and A39R complexed with the Semaphorin-binding module of PlexinC1. 20727575 0 22 from A39R 84:87 arg1 basis 11:15 Structural basis 0:15 Structural basis of semaphorin-plexin recognition 0:48 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 0 22 from A39R 84:87 arg1 mimicry 60:66 viral mimicry 54:66 viral mimicry from Sema7A and A39R 54:87 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 4 23 theme PlexinC1 605:612 arg1 propellers 619:628 the Semaphorin and PlexinC1 beta propellers 586:628 propellers 619:628 Both structures show two PlexinC1 molecules symmetrically bridged by Semaphorin dimers, in which the Semaphorin and PlexinC1 beta propellers interact in an edge-on, orthogonal orientation. 20727575 5 24 theme loop 759:762 arg1 insertion 723:731 the insertion 719:731 the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller 719:818 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 6 25 theme Sema7A 928:933 arg1 complex 935:941 the mammalian Sema7A complex 914:941 the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1 914:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 7 26 theme complex 1030:1036 arg1 structures 1038:1047 The complex structures 1026:1047 The complex structures 1026:1047 The complex structures support a conserved Semaphorin-Plexin recognition mode and suggest that Plexins are activated by dimerization. 20727575 5 27 theme PlexinC1 801:808 arg1 propeller 810:818 the PlexinC1 propeller 797:818 the PlexinC1 propeller 797:818 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 1 28 dep development 180:190 arg1 the 176:178 the 176:178 the 176:178 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 3 29 theme Sema7A 415:420 arg1 structures 401:410 the structures 397:410 the structures of Sema7A and A39R complexed with the Semaphorin-binding module of PlexinC1 397:486 We report the structures of Sema7A and A39R complexed with the Semaphorin-binding module of PlexinC1. 20727575 0 30 theme recognition 38:48 arg1 basis 11:15 Structural basis 0:15 Structural basis of semaphorin-plexin recognition 0:48 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 0 30 theme recognition 38:48 arg1 mimicry 60:66 viral mimicry 54:66 viral mimicry from Sema7A and A39R 54:87 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 6 31 theme mammalian 918:926 arg1 complex 935:941 the mammalian Sema7A complex 914:941 the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1 914:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 4 32 theme Semaphorin 558:567 arg1 dimers 569:574 Semaphorin dimers 558:574 Semaphorin dimers 558:574 Both structures show two PlexinC1 molecules symmetrically bridged by Semaphorin dimers, in which the Semaphorin and PlexinC1 beta propellers interact in an edge-on, orthogonal orientation. 20727575 0 33 from Sema7A 73:78 arg1 basis 11:15 Structural basis 0:15 Structural basis of semaphorin-plexin recognition 0:48 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 0 33 from Sema7A 73:78 arg1 mimicry 60:66 viral mimicry 54:66 viral mimicry from Sema7A and A39R 54:87 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 5 34 theme propeller 810:818 arg1 blade 786:790 blade 3 786:792 blade 3 of the PlexinC1 propeller 786:818 Both binding interfaces are dominated by the insertion of the Semaphorin's 4c-4d loop into a deep groove in blade 3 of the PlexinC1 propeller. 20727575 1 35 theme nervous 215:221 arg1 development 180:190 development 180:190 development 180:190 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 1 35 theme nervous 215:221 arg1 homeostasis 196:206 homeostasis 196:206 homeostasis 196:206 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 6 36 theme host-derived 1003:1014 arg1 PlexinC1 1016:1023 the host-derived PlexinC1 999:1023 the host-derived PlexinC1 999:1023 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 6 37 theme Sema7A 845:850 arg1 mimicry 852:858 Sema7A mimicry 845:858 Sema7A mimicry 845:858 A39R appears to achieve Sema7A mimicry by preserving key Plexin-binding determinants seen in the mammalian Sema7A complex that have evolved to achieve higher affinity binding to the host-derived PlexinC1. 20727575 1 38 theme Repulsive 114:122 arg1 signaling 124:132 Repulsive signaling 114:132 Repulsive signaling by Semaphorins and Plexins 114:159 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 0 39 theme viral 54:58 arg1 mimicry 60:66 viral mimicry 54:66 viral mimicry from Sema7A and A39R 54:87 Structural basis of semaphorin-plexin recognition and viral mimicry from Sema7A and A39R complexes with PlexinC1. 20727575 4 40 theme PlexinC1 514:521 arg1 molecules 523:531 two PlexinC1 molecules 510:531 two PlexinC1 molecules 510:531 Both structures show two PlexinC1 molecules symmetrically bridged by Semaphorin dimers, in which the Semaphorin and PlexinC1 beta propellers interact in an edge-on, orthogonal orientation. 20727575 2 41 dep immune 283:288 arg1 an 280:281 an 280:281 an 280:281 Sema7A acts as both an immune and a neural Semaphorin through PlexinC1, and A39R is a Sema7A mimic secreted by smallpox virus. 20727575 2 42 dep Sema7A 346:351 arg1 mimic 353:357 mimic 353:357 mimic 353:357 Sema7A acts as both an immune and a neural Semaphorin through PlexinC1, and A39R is a Sema7A mimic secreted by smallpox virus. 20727575 1 43 theme immune 224:229 arg1 development 180:190 development 180:190 development 180:190 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 1 43 theme immune 224:229 arg1 homeostasis 196:206 homeostasis 196:206 homeostasis 196:206 Repulsive signaling by Semaphorins and Plexins is crucial for the development and homeostasis of the nervous, immune, and cardiovascular systems. 20727575 2 44 theme smallpox 371:378 arg1 virus 380:384 smallpox virus 371:384 smallpox virus 371:384 Sema7A acts as both an immune and a neural Semaphorin through PlexinC1, and A39R is a Sema7A mimic secreted by smallpox virus. 20727575 4 45 theme beta 614:617 arg1 propellers 619:628 the Semaphorin and PlexinC1 beta propellers 586:628 propellers 619:628 Both structures show two PlexinC1 molecules symmetrically bridged by Semaphorin dimers, in which the Semaphorin and PlexinC1 beta propellers interact in an edge-on, orthogonal orientation. 20727575 4 46 theme orthogonal 654:663 arg1 orientation 665:675 an edge-on, orthogonal orientation 642:675 orientation 665:675 Both structures show two PlexinC1 molecules symmetrically bridged by Semaphorin dimers, in which the Semaphorin and PlexinC1 beta propellers interact in an edge-on, orthogonal orientation. 21886772 1 0 theme located 316:322 arg1 gene 304:307 the FuKutin-Related Protein (FKRP) gene 269:307 the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3) 269:349 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 0 theme located 316:322 arg1 FKRP 310:313 FKRP 310:313 FKRP 310:313 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 7 1 theme FKRP 1240:1243 arg1 homodimer 1245:1253 The FKRP homodimer 1236:1253 The FKRP homodimer 1236:1253 The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. 21886772 0 2 theme fibres 74:79 arg1 cisternae 45:53 the Golgi cisternae 35:53 the Golgi cisternae of skeletal muscle fibres 35:79 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 1 3 theme type 181:184 arg1 LGMD2I 190:195 LGMD2I 190:195 LGMD2I 190:195 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 3 theme type 181:184 arg1 2I 186:187 type 2I 181:187 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 5 4 from myofibrils 946:955 arg1 fibres 988:993 human rectus femoris muscle fibres 960:993 human rectus femoris muscle fibres 960:993 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 0 5 theme disulfide-linked 91:106 arg1 homodimers 108:117 disulfide-linked homodimers 91:117 disulfide-linked homodimers 91:117 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 5 6 theme muscle 981:986 arg1 fibres 988:993 human rectus femoris muscle fibres 960:993 human rectus femoris muscle fibres 960:993 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 6 7 theme protein 1186:1192 arg1 complexes 1194:1202 large multimeric protein complexes 1169:1202 large multimeric protein complexes when expressed in cell culture 1169:1233 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 8 8 theme high 1368:1371 arg1 mannose 1373:1379 high mannose 1368:1379 high mannose 1368:1379 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 2 9 theme Eye 475:477 arg1 MEB 494:496 MEB 494:496 MEB 494:496 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 2 9 theme Eye 475:477 arg1 disease 485:491 Muscle Eye Brain disease 468:491 Muscle Eye Brain disease (MEB) 468:497 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 9 10 theme Golgi 1552:1556 arg1 protein 1589:1595 a Golgi resident type II transmembrane protein 1550:1595 a Golgi resident type II transmembrane protein 1550:1595 We propose a model for FKRP which is consistent with that of a Golgi resident type II transmembrane protein. 21886772 6 11 theme multimeric 1175:1184 arg1 complexes 1194:1202 large multimeric protein complexes 1169:1202 large multimeric protein complexes when expressed in cell culture 1169:1233 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 5 12 theme marker 920:925 arg1 MG160 927:931 the middle-to-trans-Golgi marker MG160 894:931 the middle-to-trans-Golgi marker MG160 894:931 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 2 13 theme Muscle 468:473 arg1 MEB 494:496 MEB 494:496 MEB 494:496 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 2 13 theme Muscle 468:473 arg1 disease 485:491 Muscle Eye Brain disease 468:491 Muscle Eye Brain disease (MEB) 468:497 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 4 14 theme biological 696:705 arg1 function 707:714 biological function 696:714 biological function 696:714 However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. 21886772 5 15 theme human 960:964 arg1 femoris 973:979 human rectus femoris 960:979 human rectus femoris muscle fibres 960:993 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 5 16 theme sections 842:849 arg1 microscopy 806:815 immunogold electron microscopy 786:815 immunogold electron microscopy of human skeletal muscle sections 786:849 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 5 17 theme skeletal 826:833 arg1 sections 842:849 human skeletal muscle sections 820:849 human skeletal muscle sections 820:849 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 8 18 gly N-glycosylation 1415:1429 arg1 FKRP 1451:1454 FKRP homodimer 1451:1464 FKRP homodimer 1451:1464 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 8 18 gly N-glycosylation 1415:1429 arg1 multimer 1469:1476 multimer formation 1469:1486 multimer formation 1469:1486 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 9 19 with consistent 1526:1535 arg1 that 1542:1545 that 1542:1545 that 1542:1545 We propose a model for FKRP which is consistent with that of a Golgi resident type II transmembrane protein. 21886772 2 20 theme Walker-Warburg 434:447 arg1 WWS 459:461 WWS 459:461 WWS 459:461 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 2 20 theme Walker-Warburg 434:447 arg1 Syndrome 449:456 Walker-Warburg Syndrome 434:456 Walker-Warburg Syndrome (WWS) 434:462 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 5 21 theme rectus 966:971 arg1 femoris 973:979 human rectus femoris 960:979 human rectus femoris muscle fibres 960:993 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 8 22 contain contains 1347:1354 arg1 FKRP 1342:1345 FKRP 1342:1345 FKRP 1342:1345 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 8 22 contain contains 1347:1354 arg2 N-glycan 1356:1363 N-glycan 1356:1363 N-glycan of high mannose and/or hybrid type 1356:1398 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 5 23 theme muscle 835:840 arg1 sections 842:849 human skeletal muscle sections 820:849 human skeletal muscle sections 820:849 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 5 24 theme human 820:824 arg1 sections 842:849 human skeletal muscle sections 820:849 human skeletal muscle sections 820:849 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 1 25 dep Dystrophy 171:179 arg1 LGMD2I 190:195 LGMD2I 190:195 LGMD2I 190:195 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 25 dep Dystrophy 171:179 arg1 2I 186:187 type 2I 181:187 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 9 26 theme transmembrane 1575:1587 arg1 protein 1589:1595 a Golgi resident type II transmembrane protein 1550:1595 a Golgi resident type II transmembrane protein 1550:1595 We propose a model for FKRP which is consistent with that of a Golgi resident type II transmembrane protein. 21886772 3 27 gly O-glycosylation 560:574 arg1 common 530:535 common 530:535 common 530:535 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 3 27 gly O-glycosylation 560:574 arg1 α-dystroglycan 627:640 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 0 28 theme Fukutin-related 0:14 arg1 protein 16:22 Fukutin-related protein 0:22 Fukutin-related protein 0:22 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 1 29 theme FuKutin-Related 273:287 arg1 gene 304:307 the FuKutin-Related Protein (FKRP) gene 269:307 the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3) 269:349 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 29 theme FuKutin-Related 273:287 arg1 FKRP 310:313 FKRP 310:313 FKRP 310:313 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 7 30 theme N-terminal 1315:1324 arg1 Cys6 1336:1339 Cys6 1336:1339 Cys6 1336:1339 The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. 21886772 7 30 theme N-terminal 1315:1324 arg1 cysteine 1326:1333 the most N-terminal cysteine 1306:1333 the most N-terminal cysteine 1306:1333 The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. 21886772 2 31 theme Muscular 406:413 arg1 Dystrophy 415:423 Congenital Muscular Dystrophy 395:423 Congenital Muscular Dystrophy (MDC1C) 395:431 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 2 31 theme Muscular 406:413 arg1 MDC1C 426:430 MDC1C 426:430 MDC1C 426:430 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 1 32 theme Protein 289:295 arg1 gene 304:307 the FuKutin-Related Protein (FKRP) gene 269:307 the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3) 269:349 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 32 theme Protein 289:295 arg1 FKRP 310:313 FKRP 310:313 FKRP 310:313 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 0 33 theme N-terminal 126:135 arg1 interaction 137:147 an N-terminal interaction 123:147 an N-terminal interaction 123:147 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 8 34 theme type 1395:1398 arg1 N-glycan 1356:1363 N-glycan 1356:1363 N-glycan of high mannose and/or hybrid type 1356:1398 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 8 35 theme mannose 1373:1379 arg1 N-glycan 1356:1363 N-glycan 1356:1363 N-glycan of high mannose and/or hybrid type 1356:1398 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 9 36 theme resident 1558:1565 arg1 protein 1589:1595 a Golgi resident type II transmembrane protein 1550:1595 a Golgi resident type II transmembrane protein 1550:1595 We propose a model for FKRP which is consistent with that of a Golgi resident type II transmembrane protein. 21886772 5 37 theme electron 797:804 arg1 microscopy 806:815 immunogold electron microscopy 786:815 immunogold electron microscopy of human skeletal muscle sections 786:849 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 2 38 theme Congenital 395:404 arg1 Dystrophy 415:423 Congenital Muscular Dystrophy 395:423 Congenital Muscular Dystrophy (MDC1C) 395:431 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 2 38 theme Congenital 395:404 arg1 MDC1C 426:430 MDC1C 426:430 MDC1C 426:430 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 6 39 theme large 1169:1173 arg1 complexes 1194:1202 large multimeric protein complexes 1169:1202 large multimeric protein complexes when expressed in cell culture 1169:1233 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 4 40 theme further 652:658 arg1 knowledge 660:668 further knowledge 652:668 further knowledge on the FKRP structure and biological function 652:714 However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. 21886772 6 41 theme yeast 1052:1056 arg1 2-hybrid 1058:1065 pairwise yeast 2-hybrid 1043:1065 pairwise yeast 2-hybrid experiments 1043:1077 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 0 42 theme Golgi 39:43 arg1 cisternae 45:53 the Golgi cisternae 35:53 the Golgi cisternae of skeletal muscle fibres 35:79 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 1 43 theme FKRP 298:301 arg1 gene 304:307 the FuKutin-Related Protein (FKRP) gene 269:307 the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3) 269:349 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 43 theme FKRP 298:301 arg1 FKRP 310:313 FKRP 310:313 FKRP 310:313 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 6 44 theme pairwise 1043:1050 arg1 2-hybrid 1058:1065 pairwise yeast 2-hybrid 1043:1065 pairwise yeast 2-hybrid experiments 1043:1077 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 8 45 theme hybrid 1388:1393 arg1 type 1395:1398 hybrid type 1388:1398 hybrid type 1388:1398 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 1 46 theme inheritable 204:214 arg1 disorder 237:244 an inheritable autosomal, recessive disorder 201:244 disorder 237:244 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 46 theme inheritable 204:214 arg1 Dystrophy 171:179 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 6 47 theme co-immune 1084:1092 arg1 precipitation 1094:1106 co-immune precipitation 1084:1106 co-immune precipitation 1084:1106 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 1 48 from mutations 256:264 arg1 gene 304:307 the FuKutin-Related Protein (FKRP) gene 269:307 the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3) 269:349 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 48 from mutations 256:264 arg1 FKRP 310:313 FKRP 310:313 FKRP 310:313 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 3 49 theme membrane/extracellular 583:604 arg1 ECM 614:616 ECM 614:616 ECM 614:616 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 3 49 theme membrane/extracellular 583:604 arg1 matrix 606:611 membrane/extracellular matrix 583:611 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 9 50 theme type 1567:1570 arg1 protein 1589:1595 a Golgi resident type II transmembrane protein 1550:1595 a Golgi resident type II transmembrane protein 1550:1595 We propose a model for FKRP which is consistent with that of a Golgi resident type II transmembrane protein. 21886772 2 51 from Mutations 352:360 arg1 FKRP 365:368 FKRP 365:368 FKRP 365:368 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 1 52 theme autosomal 216:224 arg1 disorder 237:244 an inheritable autosomal, recessive disorder 201:244 disorder 237:244 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 52 theme autosomal 216:224 arg1 Dystrophy 171:179 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 2 53 theme Brain 479:483 arg1 MEB 494:496 MEB 494:496 MEB 494:496 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 2 53 theme Brain 479:483 arg1 disease 485:491 Muscle Eye Brain disease 468:491 Muscle Eye Brain disease (MEB) 468:497 Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). 21886772 3 54 theme matrix 606:611 arg1 α-dystroglycan 627:640 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 3 55 theme α-dystroglycan 627:640 arg1 O-glycosylation 560:574 an incomplete/aberrant O-glycosylation 537:574 an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan 537:640 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 6 56 theme cell 1222:1225 arg1 culture 1227:1233 cell culture 1222:1233 cell culture 1222:1233 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 7 57 theme disulfide 1277:1285 arg1 bridge 1287:1292 a disulfide bridge 1275:1292 a disulfide bridge provided by the most N-terminal cysteine, Cys6 1275:1339 The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. 21886772 8 58 theme FKRP 1410:1413 arg1 N-glycosylation 1415:1429 FKRP N-glycosylation 1410:1429 FKRP N-glycosylation 1410:1429 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 8 59 theme FKRP 1451:1454 arg1 homodimer 1456:1464 FKRP homodimer 1451:1464 FKRP homodimer 1451:1464 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 5 60 theme femoris 973:979 arg1 fibres 988:993 human rectus femoris muscle fibres 960:993 human rectus femoris muscle fibres 960:993 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 5 61 theme middle-to-trans-Golgi 898:918 arg1 MG160 927:931 the middle-to-trans-Golgi marker MG160 894:931 the middle-to-trans-Golgi marker MG160 894:931 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 8 62 theme multimer 1469:1476 arg1 formation 1478:1486 multimer formation 1469:1486 multimer formation 1469:1486 FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. 21886772 4 63 theme intracellular 736:748 arg1 location 750:757 its intracellular location 732:757 its intracellular location 732:757 However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. 21886772 4 64 theme FKRP 677:680 arg1 structure 682:690 the FKRP structure 673:690 the FKRP structure 673:690 However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. 21886772 7 65 theme most 1310:1313 arg1 Cys6 1336:1339 Cys6 1336:1339 Cys6 1336:1339 The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. 21886772 7 65 theme most 1310:1313 arg1 cysteine 1326:1333 the most N-terminal cysteine 1306:1333 the most N-terminal cysteine 1306:1333 The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. 21886772 1 66 theme recessive 227:235 arg1 disorder 237:244 an inheritable autosomal, recessive disorder 201:244 disorder 237:244 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 66 theme recessive 227:235 arg1 Dystrophy 171:179 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 0 67 theme muscle 67:72 arg1 fibres 74:79 skeletal muscle fibres 58:79 skeletal muscle fibres 58:79 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 3 68 theme protein 619:625 arg1 α-dystroglycan 627:640 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 the membrane/extracellular matrix (ECM) protein α-dystroglycan 579:640 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 3 69 theme incomplete/aberrant 540:558 arg1 O-glycosylation 560:574 an incomplete/aberrant O-glycosylation 537:574 an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan 537:640 These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. 21886772 6 70 theme 2-hybrid 1058:1065 arg1 experiments 1067:1077 pairwise yeast 2-hybrid experiments 1043:1077 pairwise yeast 2-hybrid experiments 1043:1077 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 1 71 theme Limb-Girdle 150:160 arg1 disorder 237:244 an inheritable autosomal, recessive disorder 201:244 disorder 237:244 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 71 theme Limb-Girdle 150:160 arg1 Dystrophy 171:179 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 6 72 theme cross-linking 1005:1017 arg1 experiments 1019:1029 Chemical cross-linking experiments 996:1029 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments 996:1077 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 21886772 0 73 theme skeletal 58:65 arg1 fibres 74:79 skeletal muscle fibres 58:79 skeletal muscle fibres 58:79 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 5 74 theme immunogold 786:795 arg1 microscopy 806:815 immunogold electron microscopy 786:815 immunogold electron microscopy of human skeletal muscle sections 786:849 Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. 21886772 0 75 link disulfide-linked 91:106 arg1 homodimers 108:117 disulfide-linked homodimers 91:117 disulfide-linked homodimers 91:117 Fukutin-related protein resides in the Golgi cisternae of skeletal muscle fibres and forms disulfide-linked homodimers via an N-terminal interaction. 21886772 1 76 theme Muscular 162:169 arg1 disorder 237:244 an inheritable autosomal, recessive disorder 201:244 disorder 237:244 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 1 76 theme Muscular 162:169 arg1 Dystrophy 171:179 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) 150:196 Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). 21886772 4 77 from knowledge 660:668 arg1 function 707:714 biological function 696:714 biological function 696:714 However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. 21886772 4 77 from knowledge 660:668 arg1 structure 682:690 the FKRP structure 673:690 the FKRP structure 673:690 However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. 21886772 6 78 theme Chemical 996:1003 arg1 experiments 1019:1029 Chemical cross-linking experiments 996:1029 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments 996:1077 Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. 3877053 8 0 theme other 1053:1057 arg1 proteins 1059:1066 other proteins 1053:1066 other proteins 1053:1066 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 10 1 with homology 1521:1528 arg1 proteases 1548:1556 other serine proteases 1535:1556 other serine proteases including trypsin and many clotting factors 1535:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 10 1 with homology 1521:1528 arg1 factors 1594:1600 many clotting factors 1580:1600 many clotting factors 1580:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 10 1 with homology 1521:1528 arg1 trypsin 1568:1574 trypsin 1568:1574 trypsin 1568:1574 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 5 2 theme plasma 648:653 arg1 codon 678:682 a TGA stop codon 667:682 a TGA stop codon 667:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 2 theme plasma 648:653 arg1 XII 662:664 plasma factor XII 648:664 plasma factor XII 648:664 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 6 3 theme acid 823:826 arg1 residues 828:835 596 amino acid residues 813:835 596 amino acid residues 813:835 The cDNA sequence predicts that plasma factor XII consists of 596 amino acid residues. 3877053 4 4 theme positive 398:405 arg1 clones 407:412 Four positive clones 393:412 Four positive clones 393:412 Four positive clones were isolated that contained DNA coding for most of factor XII mRNA. 3877053 5 5 theme XII 662:664 arg1 A 741:741 A 741:741 A 741:741 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 5 theme XII 662:664 arg1 sequence 636:643 the complete amino acid sequence 612:643 the complete amino acid sequence of plasma factor XII, a TGA stop codon 612:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 5 theme XII 662:664 arg1 DNA 560:562 DNA 560:562 DNA coding for part of an amino-terminal extension 560:609 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 5 theme XII 662:664 arg1 tail 745:748 tail 745:748 tail 745:748 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 5 theme XII 662:664 arg1 region 703:708 a 3' untranslated region 685:708 a 3' untranslated region of 150 nucleotides 685:727 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 5 theme XII 662:664 arg1 poly 736:739 a poly 734:739 a poly(A) 734:742 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 6 theme sequence 487:494 arg1 analysis 496:503 DNA sequence analysis 483:503 DNA sequence analysis of these overlapping clones 483:531 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 8 7 theme sequence 1087:1094 arg1 identity 1096:1103 extensive sequence identity 1077:1103 extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies) 1077:1266 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 8 8 theme epidermal 1160:1168 arg1 region 1189:1194 the epidermal growth factor-like region 1156:1194 the epidermal growth factor-like region 1156:1194 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 10 9 theme carboxyl-terminal 1442:1458 arg1 region 1460:1465 The carboxyl-terminal region 1438:1465 The carboxyl-terminal region of factor XII 1438:1479 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 5 10 theme amino 625:629 arg1 sequence 636:643 the complete amino acid sequence 612:643 the complete amino acid sequence of plasma factor XII, a TGA stop codon 612:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 2 11 theme oligodeoxyribonucleotides 254:278 arg1 mixtures 232:239 two mixtures 228:239 two mixtures of synthetic oligodeoxyribonucleotides as probes 228:288 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 8 12 theme factor-like 1177:1187 arg1 region 1189:1194 the epidermal growth factor-like region 1156:1194 the epidermal growth factor-like region 1156:1194 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 10 13 theme considerable 1488:1499 arg1 homology 1521:1528 considerable amino acid sequence homology 1488:1528 considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors 1488:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 11 14 theme diffraction 1707:1717 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 7 15 theme XIIa 1000:1003 arg1 formation 975:983 the formation 971:983 the formation of beta-factor XIIa 971:1003 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 10 16 theme XII 1477:1479 arg1 region 1460:1465 The carboxyl-terminal region 1438:1465 The carboxyl-terminal region of factor XII 1438:1479 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 11 17 theme resolution 1690:1699 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 9 18 theme factor 1365:1370 arg1 XII 1372:1374 factor XII 1365:1374 factor XII 1365:1374 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 8 19 theme plasminogen 1133:1143 arg1 activator 1145:1153 tissue-type plasminogen activator 1121:1153 tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) 1121:1218 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 2 20 with hybridization 209:221 arg1 mixtures 232:239 two mixtures 228:239 two mixtures of synthetic oligodeoxyribonucleotides as probes 228:288 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 10 21 theme other 1535:1539 arg1 proteases 1548:1556 other serine proteases 1535:1556 other serine proteases including trypsin and many clotting factors 1535:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 10 21 theme other 1535:1539 arg1 factors 1594:1600 many clotting factors 1580:1600 many clotting factors 1580:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 10 21 theme other 1535:1539 arg1 trypsin 1568:1574 trypsin 1568:1574 trypsin 1568:1574 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 8 22 theme kringle 1204:1210 arg1 region 1212:1217 the kringle region 1200:1217 the kringle region 1200:1217 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 8 23 dep type 1248:1251 arg1 homologies 1256:1265 homologies 1256:1265 type II homologies 1248:1265 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 10 24 theme acid 1507:1510 arg1 homology 1521:1528 considerable amino acid sequence homology 1488:1528 considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors 1488:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 8 25 with identity 1096:1103 arg1 regions 1110:1116 regions 1110:1116 regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies) 1110:1266 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 5 26 contain contained 550:558 arg1 they 545:548 they 545:548 they 545:548 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 26 contain contained 550:558 arg2 poly 736:739 a poly 734:739 a poly(A) 734:742 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 26 contain contained 550:558 arg2 region 703:708 a 3' untranslated region 685:708 a 3' untranslated region of 150 nucleotides 685:727 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 26 contain contained 550:558 arg2 A 741:741 A 741:741 A 741:741 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 26 contain contained 550:558 arg2 DNA 560:562 DNA 560:562 DNA coding for part of an amino-terminal extension 560:609 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 26 contain contained 550:558 arg2 sequence 636:643 the complete amino acid sequence 612:643 the complete amino acid sequence of plasma factor XII, a TGA stop codon 612:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 27 theme stop 673:676 arg1 codon 678:682 a TGA stop codon 667:682 a TGA stop codon 667:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 27 theme stop 673:676 arg1 XII 662:664 plasma factor XII 648:664 plasma factor XII 648:664 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 28 theme overlapping 514:524 arg1 clones 526:531 these overlapping clones 508:531 these overlapping clones 508:531 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 1 29 theme tertiary 119:126 arg1 structure 128:136 the tertiary structure 115:136 the tertiary structure of beta-factor XIIa 115:156 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 8 30 theme fibronectin 1224:1234 arg1 regions 1110:1116 regions 1110:1116 regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies) 1110:1266 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 3 31 theme XIIa 345:348 arg1 oligonucleotides 297:312 These oligonucleotides 291:312 These oligonucleotides 291:312 These oligonucleotides encoded regions of beta-factor XIIa as predicted from the amino acid sequence. 3877053 3 31 theme XIIa 345:348 arg1 regions 322:328 regions 322:328 regions of beta-factor XIIa 322:348 These oligonucleotides encoded regions of beta-factor XIIa as predicted from the amino acid sequence. 3877053 11 32 theme XIIa 1649:1652 arg1 model 1628:1632 A preliminary structural model 1603:1632 A preliminary structural model of beta-factor XIIa 1603:1652 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 5 33 theme nucleotides 717:727 arg1 A 741:741 A 741:741 A 741:741 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 33 theme nucleotides 717:727 arg1 sequence 636:643 the complete amino acid sequence 612:643 the complete amino acid sequence of plasma factor XII, a TGA stop codon 612:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 33 theme nucleotides 717:727 arg1 DNA 560:562 DNA 560:562 DNA coding for part of an amino-terminal extension 560:609 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 33 theme nucleotides 717:727 arg1 tail 745:748 tail 745:748 tail 745:748 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 33 theme nucleotides 717:727 arg1 region 703:708 a 3' untranslated region 685:708 a 3' untranslated region of 150 nucleotides 685:727 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 33 theme nucleotides 717:727 arg1 poly 736:739 a poly 734:739 a poly(A) 734:742 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 11 34 theme known 1679:1683 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 5 35 theme amino-terminal 586:599 arg1 extension 601:609 an amino-terminal extension 583:609 an amino-terminal extension 583:609 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 1 36 theme XIIa 153:156 arg1 Prediction 61:70 Prediction 61:70 Prediction of the primary structure of factor XII 61:109 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 1 36 theme XIIa 153:156 arg1 structure 128:136 the tertiary structure 115:136 the tertiary structure of beta-factor XIIa 115:156 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 5 37 theme untranslated 690:701 arg1 region 703:708 a 3' untranslated region 685:708 a 3' untranslated region of 150 nucleotides 685:727 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 4 38 theme XII 473:475 arg1 mRNA 477:480 factor XII mRNA 466:480 factor XII mRNA 466:480 Four positive clones were isolated that contained DNA coding for most of factor XII mRNA. 3877053 7 39 theme amino 859:863 arg1 sequence 870:877 the predicted amino acid sequence 845:877 the predicted amino acid sequence of factor XII 845:891 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 10 40 theme serine 1541:1546 arg1 proteases 1548:1556 other serine proteases 1535:1556 other serine proteases including trypsin and many clotting factors 1535:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 10 40 theme serine 1541:1546 arg1 factors 1594:1600 many clotting factors 1580:1600 many clotting factors 1580:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 10 40 theme serine 1541:1546 arg1 trypsin 1568:1574 trypsin 1568:1574 trypsin 1568:1574 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 1 41 theme structure 87:95 arg1 Prediction 61:70 Prediction 61:70 Prediction of the primary structure of factor XII 61:109 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 1 41 theme structure 87:95 arg1 structure 128:136 the tertiary structure 115:136 the tertiary structure of beta-factor XIIa 115:156 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 11 42 theme preliminary 1605:1615 arg1 model 1628:1632 A preliminary structural model 1603:1632 A preliminary structural model of beta-factor XIIa 1603:1652 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 3 43 theme acid 378:381 arg1 sequence 383:390 the amino acid sequence 368:390 the amino acid sequence 368:390 These oligonucleotides encoded regions of beta-factor XIIa as predicted from the amino acid sequence. 3877053 10 44 theme clotting 1585:1592 arg1 factors 1594:1600 many clotting factors 1580:1600 many clotting factors 1580:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 0 45 theme human 20:24 arg1 cDNA 55:58 human blood coagulation factor XII cDNA 20:58 human blood coagulation factor XII cDNA 20:58 Characterization of human blood coagulation factor XII cDNA. 3877053 1 46 theme factor 100:105 arg1 XII 107:109 factor XII 100:109 factor XII 100:109 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 2 47 theme human 161:165 arg1 library 178:184 A human liver cDNA library 159:184 A human liver cDNA library 159:184 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 0 48 theme coagulation 32:42 arg1 cDNA 55:58 human blood coagulation factor XII cDNA 20:58 human blood coagulation factor XII cDNA 20:58 Characterization of human blood coagulation factor XII cDNA. 3877053 6 49 theme plasma 783:788 arg1 XII 797:799 plasma factor XII 783:799 plasma factor XII 783:799 The cDNA sequence predicts that plasma factor XII consists of 596 amino acid residues. 3877053 11 50 theme elastase 1760:1767 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 0 51 theme XII 51:53 arg1 cDNA 55:58 human blood coagulation factor XII cDNA 20:58 human blood coagulation factor XII cDNA 20:58 Characterization of human blood coagulation factor XII cDNA. 3877053 8 52 with Comparison 1006:1015 arg1 proteins 1059:1066 other proteins 1053:1066 other proteins 1053:1066 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 11 53 theme beta-factor 1637:1647 arg1 XIIa 1649:1652 beta-factor XIIa 1637:1652 beta-factor XIIa 1637:1652 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 9 54 theme II 1281:1282 arg1 region 1284:1289 the type II region 1272:1289 the type II region of fibronectin 1272:1304 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 8 55 theme structure 1024:1032 arg1 Comparison 1006:1015 Comparison 1006:1015 Comparison of the structure of factor XII with other proteins 1006:1066 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 6 56 theme cDNA 755:758 arg1 sequence 760:767 The cDNA sequence 751:767 The cDNA sequence 751:767 The cDNA sequence predicts that plasma factor XII consists of 596 amino acid residues. 3877053 2 57 theme cDNA 173:176 arg1 library 178:184 A human liver cDNA library 159:184 A human liver cDNA library 159:184 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 8 58 theme factor 1037:1042 arg1 XII 1044:1046 factor XII 1037:1046 factor XII 1037:1046 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 5 59 theme extension 601:609 arg1 part 575:578 part 575:578 part of an amino-terminal extension 575:609 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 7 60 theme beta-factor 988:998 arg1 XIIa 1000:1003 beta-factor XIIa 988:1003 beta-factor XIIa 988:1003 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 10 61 theme factor 1470:1475 arg1 XII 1477:1479 factor XII 1470:1479 factor XII 1470:1479 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 5 62 theme factor 655:660 arg1 codon 678:682 a TGA stop codon 667:682 a TGA stop codon 667:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 62 theme factor 655:660 arg1 XII 662:664 plasma factor XII 648:664 plasma factor XII 648:664 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 9 63 theme collagen-binding 1317:1332 arg1 site 1334:1337 a collagen-binding site 1315:1337 a collagen-binding site 1315:1337 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 6 64 theme amino 817:821 arg1 residues 828:835 596 amino acid residues 813:835 596 amino acid residues 813:835 The cDNA sequence predicts that plasma factor XII consists of 596 amino acid residues. 3877053 9 65 theme factor 1414:1419 arg1 XII 1421:1423 factor XII 1414:1423 factor XII 1414:1423 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 5 66 theme complete 616:623 arg1 sequence 636:643 the complete amino acid sequence 612:643 the complete amino acid sequence of plasma factor XII, a TGA stop codon 612:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 8 67 theme extensive 1077:1085 arg1 identity 1096:1103 extensive sequence identity 1077:1103 extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies) 1077:1266 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 10 68 theme amino 1501:1505 arg1 homology 1521:1528 considerable amino acid sequence homology 1488:1528 considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors 1488:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 5 69 theme acid 631:634 arg1 sequence 636:643 the complete amino acid sequence 612:643 the complete amino acid sequence of plasma factor XII, a TGA stop codon 612:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 9 70 theme homologous 1344:1353 arg1 region 1355:1360 the homologous region 1340:1360 the homologous region in factor XII 1340:1374 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 9 70 theme homologous 1344:1353 arg1 responsible 1383:1393 responsible 1383:1393 responsible 1383:1393 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 8 71 theme growth 1170:1175 arg1 region 1189:1194 the epidermal growth factor-like region 1156:1194 the epidermal growth factor-like region 1156:1194 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 9 72 contain contains 1306:1313 arg2 site 1334:1337 a collagen-binding site 1315:1337 a collagen-binding site 1315:1337 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 9 72 contain contains 1306:1313 arg1 region 1284:1289 the type II region 1272:1289 the type II region of fibronectin 1272:1304 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 7 73 theme factor 882:887 arg1 XII 889:891 factor XII 882:891 factor XII 882:891 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 9 74 theme XII 1421:1423 arg1 binding 1403:1409 the binding 1399:1409 the binding of factor XII to collagen 1399:1435 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 11 75 theme x-ray 1701:1705 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 5 76 theme DNA 483:485 arg1 analysis 496:503 DNA sequence analysis 483:503 DNA sequence analysis of these overlapping clones 483:531 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 8 77 theme tissue-type 1121:1131 arg1 activator 1145:1153 tissue-type plasminogen activator 1121:1153 tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) 1121:1218 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 4 78 theme factor 466:471 arg1 mRNA 477:480 factor XII mRNA 466:480 factor XII mRNA 466:480 Four positive clones were isolated that contained DNA coding for most of factor XII mRNA. 3877053 10 79 theme sequence 1512:1519 arg1 homology 1521:1528 considerable amino acid sequence homology 1488:1528 considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors 1488:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 11 80 theme chymotrypsin 1742:1753 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 2 81 theme colony 202:207 arg1 hybridization 209:221 colony hybridization 202:221 colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes 202:288 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 1 82 theme XII 107:109 arg1 structure 87:95 the primary structure 75:95 the primary structure of factor XII 75:109 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 8 83 theme activator 1145:1153 arg1 regions 1110:1116 regions 1110:1116 regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies) 1110:1266 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 11 84 theme trypsin 1733:1739 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 5 85 theme TGA 669:671 arg1 codon 678:682 a TGA stop codon 667:682 a TGA stop codon 667:682 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 5 85 theme TGA 669:671 arg1 XII 662:664 plasma factor XII 648:664 plasma factor XII 648:664 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 2 86 theme synthetic 244:252 arg1 oligodeoxyribonucleotides 254:278 synthetic oligodeoxyribonucleotides 244:278 synthetic oligodeoxyribonucleotides as probes 244:288 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 7 87 theme peptide 919:925 arg1 bonds 927:931 three peptide bonds 913:931 three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa 913:1003 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 5 88 theme clones 526:531 arg1 analysis 496:503 DNA sequence analysis 483:503 DNA sequence analysis of these overlapping clones 483:531 DNA sequence analysis of these overlapping clones showed that they contained DNA coding for part of an amino-terminal extension, the complete amino acid sequence of plasma factor XII, a TGA stop codon, a 3' untranslated region of 150 nucleotides, and a poly(A)+ tail. 3877053 1 89 theme beta-factor 141:151 arg1 XIIa 153:156 beta-factor XIIa 141:156 beta-factor XIIa 141:156 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 3 90 theme beta-factor 333:343 arg1 XIIa 345:348 beta-factor XIIa 333:348 beta-factor XIIa 333:348 These oligonucleotides encoded regions of beta-factor XIIa as predicted from the amino acid sequence. 3877053 11 91 theme high 1685:1688 arg1 structures 1719:1728 the known high resolution x-ray diffraction structures 1675:1728 the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase 1675:1767 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 7 92 theme XII 889:891 arg1 sequence 870:877 the predicted amino acid sequence 845:877 the predicted amino acid sequence of factor XII 845:891 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 3 93 theme amino 372:376 arg1 sequence 383:390 the amino acid sequence 368:390 the amino acid sequence 368:390 These oligonucleotides encoded regions of beta-factor XIIa as predicted from the amino acid sequence. 3877053 7 94 theme acid 865:868 arg1 sequence 870:877 the predicted amino acid sequence 845:877 the predicted amino acid sequence of factor XII 845:891 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 1 95 theme primary 79:85 arg1 structure 87:95 the primary structure 75:95 the primary structure of factor XII 75:109 Prediction of the primary structure of factor XII and the tertiary structure of beta-factor XIIa. 3877053 4 96 contain contained 433:441 arg1 that 428:431 that 428:431 that 428:431 Four positive clones were isolated that contained DNA coding for most of factor XII mRNA. 3877053 4 96 contain contained 433:441 arg2 DNA 443:445 DNA 443:445 DNA coding for most of factor XII mRNA 443:480 Four positive clones were isolated that contained DNA coding for most of factor XII mRNA. 3877053 0 97 theme blood 26:30 arg1 cDNA 55:58 human blood coagulation factor XII cDNA 20:58 human blood coagulation factor XII cDNA 20:58 Characterization of human blood coagulation factor XII cDNA. 3877053 7 98 theme predicted 849:857 arg1 sequence 870:877 the predicted amino acid sequence 845:877 the predicted amino acid sequence of factor XII 845:891 Within the predicted amino acid sequence of factor XII, we have identified three peptide bonds that are cleaved by kallikrein during the formation of beta-factor XIIa. 3877053 8 99 dep activator 1145:1153 arg1 region 1212:1217 the kringle region 1200:1217 the kringle region 1200:1217 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 8 99 dep activator 1145:1153 arg1 region 1189:1194 the epidermal growth factor-like region 1156:1194 the epidermal growth factor-like region 1156:1194 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 2 100 theme liver 167:171 arg1 library 178:184 A human liver cDNA library 159:184 A human liver cDNA library 159:184 A human liver cDNA library was screened by colony hybridization with two mixtures of synthetic oligodeoxyribonucleotides as probes. 3877053 10 101 theme many 1580:1583 arg1 factors 1594:1600 many clotting factors 1580:1600 many clotting factors 1580:1600 The carboxyl-terminal region of factor XII shares considerable amino acid sequence homology with other serine proteases including trypsin and many clotting factors. 3877053 0 102 theme factor 44:49 arg1 cDNA 55:58 human blood coagulation factor XII cDNA 20:58 human blood coagulation factor XII cDNA 20:58 Characterization of human blood coagulation factor XII cDNA. 3877053 6 103 theme factor 790:795 arg1 XII 797:799 plasma factor XII 783:799 plasma factor XII 783:799 The cDNA sequence predicts that plasma factor XII consists of 596 amino acid residues. 3877053 0 104 theme cDNA 55:58 arg1 Characterization 0:15 Characterization 0:15 Characterization of human blood coagulation factor XII cDNA. 0:59 Characterization of human blood coagulation factor XII cDNA. 3877053 9 105 theme type 1276:1279 arg1 region 1284:1289 the type II region 1272:1289 the type II region of fibronectin 1272:1304 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 8 106 dep fibronectin 1224:1234 arg1 type 1237:1240 type I 1237:1242 type I 1237:1242 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 8 106 dep fibronectin 1224:1234 arg1 type 1248:1251 type II homologies 1248:1265 type II homologies 1248:1265 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 11 107 theme structural 1617:1626 arg1 model 1628:1632 A preliminary structural model 1603:1632 A preliminary structural model of beta-factor XIIa 1603:1652 A preliminary structural model of beta-factor XIIa is proposed based on the known high resolution x-ray diffraction structures of trypsin, chymotrypsin, and elastase. 3877053 8 108 theme XII 1044:1046 arg1 structure 1024:1032 the structure 1020:1032 the structure of factor XII 1020:1046 Comparison of the structure of factor XII with other proteins revealed extensive sequence identity with regions of tissue-type plasminogen activator (the epidermal growth factor-like region and the kringle region) and fibronectin (type I and type II homologies). 3877053 9 109 from region 1355:1360 arg1 XII 1372:1374 factor XII 1365:1374 factor XII 1365:1374 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 3877053 9 110 theme fibronectin 1294:1304 arg1 region 1284:1289 the type II region 1272:1289 the type II region of fibronectin 1272:1304 As the type II region of fibronectin contains a collagen-binding site, the homologous region in factor XII may be responsible for the binding of factor XII to collagen. 15308636 0 0 theme brefeldin 90:98 arg1 cells 116:120 brefeldin A-treated HepG2 cells 90:120 brefeldin A-treated HepG2 cells 90:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 1 1 theme selective 341:349 arg1 transport 351:359 membrane traffic and selective transport 320:359 membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi 320:460 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 0 2 theme human 185:189 arg1 Erv46 191:195 human Erv46 185:195 human Erv46 185:195 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 5 3 theme COPII 1097:1101 arg1 vesicles 1103:1110 COPII vesicles 1097:1110 COPII vesicles of yeast 1097:1119 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 7 4 theme mistargeting 1353:1364 arg1 experiments 1366:1376 mistargeting experiments 1353:1376 mistargeting experiments 1353:1376 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 5 5 theme uncharacterized 932:946 arg1 proteins 948:955 the uncharacterized proteins 928:955 the uncharacterized proteins 928:955 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 7 6 theme covalent 1326:1333 arg1 cross-linking 1335:1347 covalent cross-linking 1326:1347 covalent cross-linking 1326:1347 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 5 7 theme yeast 1115:1119 arg1 vesicles 1103:1110 COPII vesicles 1097:1110 COPII vesicles of yeast 1097:1119 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 4 8 theme cargo 840:844 arg1 receptors 846:854 established and putative cargo receptors 815:854 established and putative cargo receptors 815:854 Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. 15308636 2 9 theme HepG2 578:582 arg1 cells 584:588 HepG2 cells 578:588 HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 578:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 0 10 theme HepG2 110:114 arg1 cells 116:120 brefeldin A-treated HepG2 cells 90:120 brefeldin A-treated HepG2 cells 90:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 11 from Proteomics 0:9 arg1 cells 116:120 brefeldin A-treated HepG2 cells 90:120 brefeldin A-treated HepG2 cells 90:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 2 12 theme membranes 563:571 arg1 purification 541:552 the purification 537:552 the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 537:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 7 13 theme small 1408:1412 arg1 RNAs 1426:1429 small interfering RNAs 1408:1429 small interfering RNAs 1408:1429 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 6 14 theme Erv46p 1218:1223 arg1 homologs 1195:1202 the human homologs 1185:1202 the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi 1185:1263 ERGIC-32 localizes to the ERGIC and partially colocalizes with the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi. 15308636 0 15 theme A-treated 100:108 arg1 cells 116:120 brefeldin A-treated HepG2 cells 90:120 brefeldin A-treated HepG2 cells 90:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 1 16 theme cargo 364:368 arg1 transport 351:359 membrane traffic and selective transport 320:359 membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi 320:460 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 2 17 theme ERGIC 557:561 arg1 membranes 563:571 ERGIC membranes 557:571 ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 557:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 5 18 theme 32-kDa 960:965 arg1 protein 1019:1025 a novel cycling membrane protein 994:1025 a novel cycling membrane protein with sequence homology to Erv41p and Erv46p 994:1069 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 18 theme 32-kDa 960:965 arg1 protein 967:973 a 32-kDa protein 958:973 a 32-kDa protein termed ERGIC-32 958:989 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 2 19 from purification 541:552 arg1 cells 584:588 HepG2 cells 578:588 HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 578:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 2 20 theme brefeldin 603:611 arg1 A 613:613 brefeldin A 603:613 brefeldin A 603:613 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 2 20 theme brefeldin 603:611 arg1 drug 618:621 a drug 616:621 a drug known to accumulate cycling proteins in the ERGIC 616:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 7 21 theme ERGIC-32 1396:1403 arg1 silencing 1383:1391 silencing 1383:1391 silencing of ERGIC-32 by small interfering RNAs 1383:1429 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 4 22 dep membrane 904:911 arg1 traffic 913:919 traffic 913:919 traffic 913:919 Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. 15308636 2 23 theme cycling 643:649 arg1 proteins 651:658 cycling proteins 643:658 cycling proteins 643:658 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 9 24 dep proteins 1689:1696 arg1 cycling 1706:1712 cycling 1706:1712 cycling 1706:1712 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 9 25 from isolation 1594:1602 arg1 cells 1634:1638 BFA-treated cells 1622:1638 BFA-treated cells 1622:1638 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 1 26 theme endoplasmic 382:392 arg1 ER 405:406 ER 405:406 ER 405:406 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 1 26 theme endoplasmic 382:392 arg1 reticulum 394:402 the endoplasmic reticulum 378:402 the endoplasmic reticulum (ER) 378:407 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 4 27 theme putative 831:838 arg1 receptors 846:854 established and putative cargo receptors 815:854 established and putative cargo receptors 815:854 Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. 15308636 2 28 theme cycling 479:485 arg1 proteins 487:494 new cycling proteins 475:494 new cycling proteins 475:494 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 9 29 theme novel 1571:1575 arg1 approach 1577:1584 Our novel approach 1567:1584 Our novel approach for the isolation of the ERGIC from BFA-treated cells 1567:1638 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 1 30 theme early 276:280 arg1 pathway 292:298 the early secretory pathway 272:298 the early secretory pathway 272:298 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 4 31 theme established 815:825 arg1 receptors 846:854 established and putative cargo receptors 815:854 established and putative cargo receptors 815:854 Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. 15308636 9 32 theme BFA-treated 1622:1632 arg1 cells 1634:1638 BFA-treated cells 1622:1638 BFA-treated cells 1622:1638 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 0 33 theme reticulum-Golgi 26:40 arg1 compartment 55:65 endoplasmic reticulum-Golgi intermediate compartment 14:65 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 14:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 33 theme reticulum-Golgi 26:40 arg1 ERGIC 68:72 ERGIC 68:72 ERGIC 68:72 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 1 34 theme secretory 282:290 arg1 pathway 292:298 the early secretory pathway 272:298 the early secretory pathway 272:298 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 5 35 theme cycling 1002:1008 arg1 protein 967:973 a 32-kDa protein 958:973 a 32-kDa protein termed ERGIC-32 958:989 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 35 theme cycling 1002:1008 arg1 proteins 1076:1083 two proteins 1072:1083 two proteins enriched in COPII vesicles of yeast 1072:1119 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 35 theme cycling 1002:1008 arg1 protein 1019:1025 a novel cycling membrane protein 994:1025 a novel cycling membrane protein with sequence homology to Erv41p and Erv46p 994:1069 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 1 36 theme Cycling 198:204 arg1 proteins 206:213 Cycling proteins 198:213 Cycling proteins 198:213 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 0 37 theme endoplasmic 14:24 arg1 compartment 55:65 endoplasmic reticulum-Golgi intermediate compartment 14:65 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 14:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 37 theme endoplasmic 14:24 arg1 ERGIC 68:72 ERGIC 68:72 ERGIC 68:72 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 1 38 theme pathway 292:298 arg1 function 260:267 function 260:267 function 260:267 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 1 38 theme pathway 292:298 arg1 organization 243:254 organization 243:254 organization 243:254 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 2 39 theme novel 517:521 arg1 procedure 523:531 a novel procedure 515:531 a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 515:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 5 40 with protein 1019:1025 arg1 homology 1041:1048 sequence homology 1032:1048 sequence homology to Erv41p and Erv46p 1032:1069 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 7 41 theme human 1290:1294 arg1 hErv46 1303:1308 hErv46 1303:1308 hErv46 1303:1308 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 7 41 theme human 1290:1294 arg1 Erv46 1296:1300 human Erv46 1290:1300 human Erv46 (hErv46) 1290:1309 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 0 42 theme compartment 55:65 arg1 membranes 75:83 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes 14:83 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 14:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 8 43 theme complex 1536:1542 arg1 ERGIC-32 1481:1488 ERGIC-32 1481:1488 ERGIC-32 1481:1488 We propose that ERGIC-32 functions as a modulator of the hErv41-hErv46 complex by stabilizing hErv46. 15308636 8 43 theme complex 1536:1542 arg1 modulator 1505:1513 a modulator 1503:1513 a modulator of the hErv41-hErv46 complex 1503:1542 We propose that ERGIC-32 functions as a modulator of the hErv41-hErv46 complex by stabilizing hErv46. 15308636 0 44 theme new 145:147 arg1 protein 157:163 a new cycling protein 143:163 a new cycling protein that interacts with human Erv46 143:195 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 44 theme new 145:147 arg1 ERGIC-32 133:140 ERGIC-32 133:140 ERGIC-32 133:140 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 1 45 dep organization 243:254 arg1 the 239:241 the 239:241 the 239:241 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 4 46 theme Major 784:788 arg1 proteins 790:797 Major proteins 784:797 Major proteins 784:797 Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. 15308636 7 47 theme interfering 1414:1424 arg1 RNAs 1426:1429 small interfering RNAs 1408:1429 small interfering RNAs 1408:1429 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 0 48 theme intermediate 42:53 arg1 compartment 55:65 endoplasmic reticulum-Golgi intermediate compartment 14:65 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 14:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 48 theme intermediate 42:53 arg1 ERGIC 68:72 ERGIC 68:72 ERGIC 68:72 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 9 49 from cells 1634:1638 arg1 isolation 1594:1602 the isolation 1590:1602 the isolation of the ERGIC from BFA-treated cells 1590:1638 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 9 49 from cells 1634:1638 arg1 ERGIC 1611:1615 the ERGIC 1607:1615 the ERGIC from BFA-treated cells 1607:1638 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 9 50 theme proteins 1689:1696 arg1 identification 1667:1680 the identification 1663:1680 the identification of all proteins rapidly cycling early in the secretory pathway 1663:1743 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 5 51 theme membrane 1010:1017 arg1 protein 967:973 a 32-kDa protein 958:973 a 32-kDa protein termed ERGIC-32 958:989 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 51 theme membrane 1010:1017 arg1 proteins 1076:1083 two proteins 1072:1083 two proteins enriched in COPII vesicles of yeast 1072:1119 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 51 theme membrane 1010:1017 arg1 protein 1019:1025 a novel cycling membrane protein 994:1025 a novel cycling membrane protein with sequence homology to Erv41p and Erv46p 994:1069 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 1 52 theme important 220:228 arg1 roles 230:234 important roles 220:234 important roles 220:234 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 2 53 from cells 584:588 arg1 purification 541:552 the purification 537:552 the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 537:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 2 53 from cells 584:588 arg1 membranes 563:571 ERGIC membranes 557:571 ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC 557:671 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 4 54 theme protein 881:887 arg1 maturation 889:898 protein maturation 881:898 protein maturation 881:898 Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. 15308636 9 55 theme secretory 1727:1735 arg1 pathway 1737:1743 the secretory pathway 1723:1743 the secretory pathway 1723:1743 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 6 56 theme Erv41p 1207:1212 arg1 homologs 1195:1202 the human homologs 1185:1202 the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi 1185:1263 ERGIC-32 localizes to the ERGIC and partially colocalizes with the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi. 15308636 7 57 theme hErv46 1457:1462 arg1 turnover 1445:1452 the turnover 1441:1452 the turnover of hErv46 1441:1462 ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. 15308636 0 58 theme cycling 149:155 arg1 protein 157:163 a new cycling protein 143:163 a new cycling protein that interacts with human Erv46 143:195 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 58 theme cycling 149:155 arg1 ERGIC-32 133:140 ERGIC-32 133:140 ERGIC-32 133:140 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 5 59 theme sequence 1032:1039 arg1 homology 1041:1048 sequence homology 1032:1048 sequence homology to Erv41p and Erv46p 1032:1069 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 9 60 dep cycling 1706:1712 arg1 early 1714:1718 early 1714:1718 early 1714:1718 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 1 61 theme traffic 329:335 arg1 transport 351:359 membrane traffic and selective transport 320:359 membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi 320:460 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 0 62 from cells 116:120 arg1 Proteomics 0:9 Proteomics 0:9 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 0:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 0 62 from cells 116:120 arg1 membranes 75:83 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes 14:83 endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 14:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 9 63 theme ERGIC 1611:1615 arg1 isolation 1594:1602 the isolation 1590:1602 the isolation of the ERGIC from BFA-treated cells 1590:1638 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 15308636 3 64 theme mass 765:768 arg1 spectrometry 770:781 mass spectrometry 765:781 mass spectrometry 765:781 Membranes enriched 110-fold over the homogenate for ERGIC-53 were obtained and analyzed by mass spectrometry. 15308636 1 65 theme intermediate 414:425 arg1 ERGIC 440:444 ERGIC 440:444 ERGIC 440:444 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 1 65 theme intermediate 414:425 arg1 compartment 427:437 the intermediate compartment 410:437 the intermediate compartment (ERGIC) 410:445 Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. 15308636 6 66 theme human 1189:1193 arg1 homologs 1195:1202 the human homologs 1185:1202 the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi 1185:1263 ERGIC-32 localizes to the ERGIC and partially colocalizes with the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi. 15308636 0 67 theme membranes 75:83 arg1 Proteomics 0:9 Proteomics 0:9 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells 0:120 Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. 15308636 2 68 theme new 475:477 arg1 proteins 487:494 new cycling proteins 475:494 new cycling proteins 475:494 To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. 15308636 5 69 theme novel 996:1000 arg1 protein 967:973 a 32-kDa protein 958:973 a 32-kDa protein termed ERGIC-32 958:989 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 69 theme novel 996:1000 arg1 proteins 1076:1083 two proteins 1072:1083 two proteins enriched in COPII vesicles of yeast 1072:1119 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 5 69 theme novel 996:1000 arg1 protein 1019:1025 a novel cycling membrane protein 994:1025 a novel cycling membrane protein with sequence homology to Erv41p and Erv46p 994:1069 Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. 15308636 9 70 from cycling 1706:1712 arg1 pathway 1737:1743 the secretory pathway 1723:1743 the secretory pathway 1723:1743 Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway. 17509700 0 0 theme amino 145:149 arg1 acids 151:155 24 amino acids 142:155 24 amino acids 142:155 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 1 theme OCTN2 615:619 arg1 loop 607:610 the first extracellular loop 583:610 the first extracellular loop of OCTN2 583:619 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 1 2 theme novel 201:205 arg1 indispensable 243:255 indispensable 243:255 indispensable 243:255 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 2 theme novel 201:205 arg1 OCTN2 234:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 5 3 from retention 967:975 arg1 ER 984:985 the ER 980:985 the ER 980:985 In addition, the retention in the ER caused no carnitine uptake into the cells. 17509700 1 4 theme subsequent 308:317 arg1 metabolism 330:339 subsequent fatty acid metabolism 308:339 subsequent fatty acid metabolism 308:339 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 6 5 theme first 1189:1193 arg1 loop 1209:1212 the first extracellular loop 1185:1212 the first extracellular loop 1185:1212 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 0 6 theme OCTN2 192:196 arg1 loop 184:187 the first extracellular loop 160:187 the first extracellular loop of OCTN2 160:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 0 7 theme acids 151:155 arg1 insertion 129:137 insertion 129:137 insertion of 24 amino acids in the first extracellular loop of OCTN2 129:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 6 8 from OCTN2 1137:1141 arg1 characteristics 1092:1106 the biochemical and functional characteristics 1061:1106 the biochemical and functional characteristics of OCTN2VT 1061:1117 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 6 8 from OCTN2 1137:1141 arg1 distinct 1123:1130 distinct 1123:1130 distinct 1123:1130 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 0 9 from insertion 129:137 arg1 loop 184:187 the first extracellular loop 160:187 the first extracellular loop of OCTN2 160:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 1 10 theme fatty 319:323 arg1 metabolism 330:339 subsequent fatty acid metabolism 308:339 subsequent fatty acid metabolism 308:339 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 6 11 theme OCTN2VT 1111:1117 arg1 characteristics 1092:1106 the biochemical and functional characteristics 1061:1106 the biochemical and functional characteristics of OCTN2VT 1061:1117 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 6 11 theme OCTN2VT 1111:1117 arg1 distinct 1123:1130 distinct 1123:1130 distinct 1123:1130 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 1 12 theme cation 215:220 arg1 indispensable 243:255 indispensable 243:255 indispensable 243:255 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 12 theme cation 215:220 arg1 OCTN2 234:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 13 theme acid 325:328 arg1 metabolism 330:339 subsequent fatty acid metabolism 308:339 subsequent fatty acid metabolism 308:339 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 14 from transport 271:279 arg1 mitochondria 348:359 the mitochondria 344:359 the mitochondria 344:359 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 15 theme transporter 222:232 arg1 indispensable 243:255 indispensable 243:255 indispensable 243:255 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 15 theme transporter 222:232 arg1 OCTN2 234:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 2 16 theme located 454:460 arg1 sequence 445:452 a 72-base-pair sequence 430:452 a 72-base-pair sequence located in the first intron of OCTN2 gene 430:494 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 4 17 gly N-glycosylation 843:857 arg1 membrane 822:829 the plasma membrane 811:829 the plasma membrane with robust N-glycosylation 811:857 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 0 18 theme first 164:168 arg1 loop 184:187 the first extracellular loop 160:187 the first extracellular loop of OCTN2 160:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 3 19 theme biochemical 730:740 arg1 characteristics 742:756 their biochemical characteristics 724:756 their biochemical characteristics 724:756 Despite the similarity between OCTN2 and OCTN2VT regarding primary structure and tissue distribution, their biochemical characteristics were significantly different. 17509700 6 20 theme amino 1170:1174 arg1 acids 1176:1180 24 amino acids 1167:1180 24 amino acids 1167:1180 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 6 21 theme functional 1081:1090 arg1 characteristics 1092:1106 the biochemical and functional characteristics 1061:1106 the biochemical and functional characteristics of OCTN2VT 1061:1117 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 6 21 theme functional 1081:1090 arg1 distinct 1123:1130 distinct 1123:1130 distinct 1123:1130 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 2 22 theme OCTN2 404:408 arg1 variant 393:399 a novel splice variant 378:399 a novel splice variant 378:399 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 23 theme OCTN2 533:537 arg1 exons 516:520 exons 1 and 2 516:528 exons 1 and 2 of OCTN2 516:537 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 6 24 theme acids 1176:1180 arg1 insertion 1154:1162 the insertion 1150:1162 the insertion of 24 amino acids in the first extracellular loop 1150:1212 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 0 25 theme extracellular 170:182 arg1 loop 184:187 the first extracellular loop 160:187 the first extracellular loop of OCTN2 160:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 26 theme acids 574:578 arg1 insertion 552:560 the insertion 548:560 the insertion of 24 amino acids in the first extracellular loop of OCTN2 548:619 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 27 from located 454:460 arg1 intron 475:480 the first intron 465:480 the first intron of OCTN2 gene 465:494 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 0 28 theme splice 11:16 arg1 OCTN2VT 0:6 OCTN2VT 0:6 OCTN2VT 0:6 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 0 28 theme splice 11:16 arg1 variant 18:24 a splice variant 9:24 a splice variant of OCTN2 9:33 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 29 theme OCTN2 485:489 arg1 gene 491:494 OCTN2 gene 485:494 OCTN2 gene 485:494 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 30 theme amino 568:572 arg1 acids 574:578 24 amino acids 565:578 24 amino acids 565:578 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 1 31 theme organic 207:213 arg1 indispensable 243:255 indispensable 243:255 indispensable 243:255 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 1 31 theme organic 207:213 arg1 OCTN2 234:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 199:238 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 2 32 theme novel 380:384 arg1 variant 393:399 a novel splice variant 378:399 a novel splice variant 378:399 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 33 theme splice 386:391 arg1 variant 393:399 a novel splice variant 378:399 a novel splice variant 378:399 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 4 34 theme poor 928:931 arg1 N-glycosylation 933:947 poor N-glycosylation 928:947 poor N-glycosylation 928:947 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 4 35 with reticulum 908:916 arg1 N-glycosylation 933:947 poor N-glycosylation 928:947 poor N-glycosylation 928:947 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 4 36 with membrane 822:829 arg1 N-glycosylation 843:857 robust N-glycosylation 836:857 robust N-glycosylation 836:857 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 6 37 theme biochemical 1065:1075 arg1 characteristics 1092:1106 the biochemical and functional characteristics 1061:1106 the biochemical and functional characteristics of OCTN2VT 1061:1117 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 6 37 theme biochemical 1065:1075 arg1 distinct 1123:1130 distinct 1123:1130 distinct 1123:1130 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 6 38 theme extracellular 1195:1207 arg1 loop 1209:1212 the first extracellular loop 1185:1212 the first extracellular loop 1185:1212 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 2 39 dep exons 516:520 arg1 2 528:528 2 528:528 2 528:528 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 39 dep exons 516:520 arg1 1 522:522 1 522:522 1 522:522 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 1 40 theme carnitine 261:269 arg1 transport 271:279 carnitine transport 261:279 carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria 261:359 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 0 41 theme endoplasmic 97:107 arg1 reticulum 109:117 the endoplasmic reticulum 93:117 the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2 93:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 42 theme first 469:473 arg1 intron 475:480 the first intron 465:480 the first intron of OCTN2 gene 465:494 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 4 43 dep expressed 798:806 arg1 whereas 860:866 whereas 860:866 whereas 860:866 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 3 44 theme primary 681:687 arg1 structure 689:697 primary structure 681:697 primary structure 681:697 Despite the similarity between OCTN2 and OCTN2VT regarding primary structure and tissue distribution, their biochemical characteristics were significantly different. 17509700 5 45 theme carnitine 997:1005 arg1 uptake 1007:1012 no carnitine uptake 994:1012 no carnitine uptake 994:1012 In addition, the retention in the ER caused no carnitine uptake into the cells. 17509700 2 46 from insertion 552:560 arg1 loop 607:610 the first extracellular loop 583:610 the first extracellular loop of OCTN2 583:619 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 0 47 theme OCTN2 29:33 arg1 OCTN2VT 0:6 OCTN2VT 0:6 OCTN2VT 0:6 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 0 47 theme OCTN2 29:33 arg1 variant 18:24 a splice variant 9:24 a splice variant of OCTN2 9:33 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 4 48 theme plasma 815:820 arg1 membrane 822:829 the plasma membrane 811:829 the plasma membrane with robust N-glycosylation 811:857 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 0 49 from retention 80:88 arg1 reticulum 109:117 the endoplasmic reticulum 93:117 the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2 93:196 OCTN2VT, a splice variant of OCTN2, does not transport carnitine because of the retention in the endoplasmic reticulum caused by insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 6 50 theme due 1143:1145 arg1 OCTN2 1137:1141 OCTN2 1137:1141 OCTN2 due to the insertion of 24 amino acids in the first extracellular loop 1137:1212 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 2 51 theme extracellular 593:605 arg1 loop 607:610 the first extracellular loop 583:610 the first extracellular loop of OCTN2 583:619 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 1 52 theme plasma 288:293 arg1 membrane 295:302 plasma membrane 288:302 plasma membrane 288:302 A novel organic cation transporter OCTN2 is indispensable for carnitine transport across plasma membrane and subsequent fatty acid metabolism in the mitochondria. 17509700 4 53 theme robust 836:841 arg1 N-glycosylation 843:857 robust N-glycosylation 836:857 robust N-glycosylation 836:857 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 2 54 theme first 587:591 arg1 loop 607:610 the first extracellular loop 583:610 the first extracellular loop of OCTN2 583:619 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 55 from intron 475:480 arg1 located 454:460 located 454:460 located 454:460 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 2 56 theme gene 491:494 arg1 intron 475:480 the first intron 465:480 the first intron of OCTN2 gene 465:494 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 6 57 from insertion 1154:1162 arg1 loop 1209:1212 the first extracellular loop 1185:1212 the first extracellular loop 1185:1212 These results demonstrate that the biochemical and functional characteristics of OCTN2VT are distinct from OCTN2 due to the insertion of 24 amino acids in the first extracellular loop. 17509700 2 58 theme 72-base-pair 432:443 arg1 sequence 445:452 a 72-base-pair sequence 430:452 a 72-base-pair sequence located in the first intron of OCTN2 gene 430:494 Here, we report a novel splice variant of OCTN2 (OCTN2VT), in which a 72-base-pair sequence located in the first intron of OCTN2 gene was spliced between exons 1 and 2 of OCTN2, causing the insertion of 24 amino acids in the first extracellular loop of OCTN2. 17509700 4 59 theme endoplasmic 896:906 arg1 reticulum 908:916 the endoplasmic reticulum 892:916 the endoplasmic reticulum (ER) with poor N-glycosylation 892:947 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 4 59 theme endoplasmic 896:906 arg1 ER 919:920 ER 919:920 ER 919:920 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 4 60 gly N-glycosylation 933:947 arg1 reticulum 908:916 the endoplasmic reticulum 892:916 the endoplasmic reticulum (ER) with poor N-glycosylation 892:947 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 4 60 gly N-glycosylation 933:947 arg1 ER 919:920 ER 919:920 ER 919:920 OCTN2 was expressed on the plasma membrane with robust N-glycosylation, whereas OCTN2VT was retained in the endoplasmic reticulum (ER) with poor N-glycosylation. 17509700 3 61 theme tissue 703:708 arg1 distribution 710:721 tissue distribution 703:721 tissue distribution 703:721 Despite the similarity between OCTN2 and OCTN2VT regarding primary structure and tissue distribution, their biochemical characteristics were significantly different. 27391701 9 0 from enriched 1537:1544 arg1 regions 1571:1577 short protein binding regions 1549:1577 short protein binding regions called Molecular Recognition Features (MoRFs) 1549:1623 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 4 1 contain have 671:674 arg2 IDR 689:691 at least one IDR 676:691 at least one IDR of 30 amino acids 676:709 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 4 1 contain have 671:674 arg1 proteins 662:669 transmembrane proteins 648:669 transmembrane proteins 648:669 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 4 1 contain have 671:674 arg2 more 714:717 more 714:717 more 714:717 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 4 1 contain have 671:674 arg1 % 643:643 50% 641:643 50% of transmembrane proteins 641:669 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 7 2 from cell 1158:1161 arg1 enriched 1146:1153 enriched 1146:1153 enriched 1146:1153 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 3 3 from prevalence 509:518 arg1 proteins 571:578 transmembrane proteins 557:578 transmembrane proteins 557:578 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 9 4 theme binding 1563:1569 arg1 regions 1571:1577 short protein binding regions 1549:1577 short protein binding regions called Molecular Recognition Features (MoRFs) 1549:1623 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 8 5 theme protein-protein 1471:1485 arg1 events 1499:1504 more protein-protein interaction events 1466:1504 more protein-protein interaction events 1466:1504 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 0 6 from Disorder 10:17 arg1 Proteins 36:43 Transmembrane Proteins 22:43 Transmembrane Proteins 22:43 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 1 7 theme amino 156:160 arg1 acids 162:166 amino acids 156:166 amino acids 156:166 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 8 8 theme translational 1351:1363 arg1 sites 1329:1333 phosphorylation sites 1313:1333 phosphorylation sites 1313:1333 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 8 8 theme translational 1351:1363 arg1 modification 1365:1376 a crucial post translational modification 1336:1376 a crucial post translational modification in signal transduction 1336:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 2 9 attach linked 325:330 arg2 IDP 252:254 IDP 252:254 IDP 252:254 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 9 attach linked 325:330 arg1 cancer 405:410 cancer 405:410 cancer 405:410 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 9 attach linked 325:330 arg1 signaling 378:386 cell signaling 373:386 cell signaling 373:386 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 9 attach linked 325:330 arg1 binding 393:399 DNA binding 389:399 DNA binding 389:399 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 9 attach linked 325:330 arg2 Proteins 242:249 Intrinsic Disorder containing Proteins 212:249 Intrinsic Disorder containing Proteins (IDP) 212:255 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 9 attach linked 325:330 arg1 processes 353:361 multiple cellular processes 335:361 multiple cellular processes including cell signaling, DNA binding and cancer 335:410 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 3 10 theme transmembrane 557:569 arg1 proteins 571:578 transmembrane proteins 557:578 transmembrane proteins 557:578 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 3 11 theme IDRs 536:539 arg1 function 524:531 function 524:531 function 524:531 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 3 11 theme IDRs 536:539 arg1 prevalence 509:518 prevalence 509:518 prevalence 509:518 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 6 12 theme successful 973:982 arg1 prediction 984:993 the successful prediction 969:993 the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117 969:1073 This was supported by the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117, as confirmed experimentally. 27391701 5 13 theme topology 915:922 arg1 algorithms 935:944 topology prediction algorithms 915:944 topology prediction algorithms 915:944 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 4 14 theme transmembrane 648:660 arg1 proteins 662:669 transmembrane proteins 648:669 transmembrane proteins 648:669 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 8 15 theme phosphorylation 1313:1327 arg1 sites 1329:1333 phosphorylation sites 1313:1333 phosphorylation sites 1313:1333 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 8 15 theme phosphorylation 1313:1327 arg1 modification 1365:1376 a crucial post translational modification 1336:1376 a crucial post translational modification in signal transduction 1336:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 5 16 theme transmembrane 822:834 arg1 proteins 836:843 multi-pass transmembrane proteins 811:843 multi-pass transmembrane proteins 811:843 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 4 17 theme proteins 662:669 arg1 proteins 662:669 transmembrane proteins 648:669 transmembrane proteins 648:669 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 4 17 theme proteins 662:669 arg1 % 643:643 50% 641:643 50% of transmembrane proteins 641:669 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 0 18 from Roles 46:50 arg1 Topology 69:76 Topology 69:76 Topology 69:76 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 0 18 from Roles 46:50 arg1 Signaling 55:63 Signaling 55:63 Signaling 55:63 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 7 19 theme Pathway 1105:1111 arg1 analysis 1113:1120 Pathway analysis 1105:1120 Pathway analysis 1105:1120 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 6 20 theme transmembrane 1045:1057 arg1 protein 1059:1065 the uncharacterized multi-pass transmembrane protein TMEM117 1014:1073 the uncharacterized multi-pass transmembrane protein TMEM117 1014:1073 This was supported by the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117, as confirmed experimentally. 27391701 1 21 theme acids 162:166 arg1 stretches 143:151 peculiar stretches 134:151 peculiar stretches of amino acids that lack stable conformations in solution 134:209 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 1 21 theme acids 162:166 arg1 regions 115:121 Intrinsically disordered regions 90:121 Intrinsically disordered regions (IDRs) 90:128 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 9 22 theme short 1549:1553 arg1 regions 1571:1577 short protein binding regions 1549:1577 short protein binding regions called Molecular Recognition Features (MoRFs) 1549:1623 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 5 23 theme proteins 836:843 arg1 side 792:795 the cytoplasmic side 776:795 the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms 776:944 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 7 24 dep cell 1158:1161 arg1 projection 1163:1172 projection 1163:1172 projection 1163:1172 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 8 25 from enriched 1301:1308 arg1 sites 1329:1333 phosphorylation sites 1313:1333 phosphorylation sites 1313:1333 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 8 25 from enriched 1301:1308 arg1 modification 1365:1376 a crucial post translational modification 1336:1376 a crucial post translational modification in signal transduction 1336:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 3 26 used used 421:424 arg2 we 418:419 we 418:419 we 418:419 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 7 27 from axons 1178:1182 arg1 enriched 1146:1153 enriched 1146:1153 enriched 1146:1153 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 0 28 theme Intrinsic 0:8 arg1 Disorder 10:17 Intrinsic Disorder 0:17 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 0:88 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 10 29 theme transmembrane 1687:1699 arg1 IDPs 1701:1704 the transmembrane IDPs 1683:1704 the transmembrane IDPs 1683:1704 Altogether our analyses strongly support the notion that the transmembrane IDPs act as hubs in cellular signal events. 27391701 10 29 theme transmembrane 1687:1699 arg1 hubs 1713:1716 hubs 1713:1716 hubs in cellular signal events 1713:1742 Altogether our analyses strongly support the notion that the transmembrane IDPs act as hubs in cellular signal events. 27391701 5 30 theme prediction 924:933 arg1 algorithms 935:944 topology prediction algorithms 915:944 topology prediction algorithms 915:944 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 2 31 theme cellular 344:351 arg1 signaling 378:386 cell signaling 373:386 cell signaling 373:386 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 31 theme cellular 344:351 arg1 processes 353:361 multiple cellular processes 335:361 multiple cellular processes including cell signaling, DNA binding and cancer 335:410 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 31 theme cellular 344:351 arg1 cancer 405:410 cancer 405:410 cancer 405:410 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 31 theme cellular 344:351 arg1 binding 393:399 DNA binding 389:399 DNA binding 389:399 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 0 32 theme Transmembrane 22:34 arg1 Proteins 36:43 Transmembrane Proteins 22:43 Transmembrane Proteins 22:43 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 5 33 dep proteins 836:843 arg1 suggesting 846:855 suggesting 846:855 suggesting that disorder prediction could increase the confidence of topology prediction algorithms 846:944 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 1 34 theme disordered 104:113 arg1 stretches 143:151 peculiar stretches 134:151 peculiar stretches of amino acids that lack stable conformations in solution 134:209 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 1 34 theme disordered 104:113 arg1 IDRs 124:127 IDRs 124:127 IDRs 124:127 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 1 34 theme disordered 104:113 arg1 regions 115:121 Intrinsically disordered regions 90:121 Intrinsically disordered regions (IDRs) 90:128 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 9 35 theme Molecular 1586:1594 arg1 Features 1608:1615 Molecular Recognition Features 1586:1615 Molecular Recognition Features (MoRFs) 1586:1623 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 9 35 theme Molecular 1586:1594 arg1 MoRFs 1618:1622 MoRFs 1618:1622 MoRFs 1618:1622 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 3 36 theme computational 426:438 arg1 analyses 440:447 computational analyses 426:447 computational analyses 426:447 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 5 37 theme cytoplasmic 780:790 arg1 side 792:795 the cytoplasmic side 776:795 the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms 776:944 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 10 38 theme signal 1730:1735 arg1 events 1737:1742 cellular signal events 1721:1742 cellular signal events 1721:1742 Altogether our analyses strongly support the notion that the transmembrane IDPs act as hubs in cellular signal events. 27391701 0 39 dep Signaling 55:63 arg1 Prediction 78:87 Prediction 78:87 Prediction 78:87 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 8 40 theme post 1346:1349 arg1 sites 1329:1333 phosphorylation sites 1313:1333 phosphorylation sites 1313:1333 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 8 40 theme post 1346:1349 arg1 modification 1365:1376 a crucial post translational modification 1336:1376 a crucial post translational modification in signal transduction 1336:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 2 41 theme DNA 389:391 arg1 binding 393:399 DNA binding 389:399 DNA binding 389:399 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 9 42 theme protein 1555:1561 arg1 regions 1571:1577 short protein binding regions 1549:1577 short protein binding regions called Molecular Recognition Features (MoRFs) 1549:1623 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 8 43 theme ordered 1425:1431 arg1 proteins 1433:1440 fully ordered proteins 1419:1440 fully ordered proteins 1419:1440 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 8 44 theme signal 1381:1386 arg1 transduction 1388:1399 signal transduction 1381:1399 signal transduction 1381:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 1 45 theme stable 178:183 arg1 conformations 185:197 stable conformations 178:197 stable conformations 178:197 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 6 46 theme multi-pass 1034:1043 arg1 protein 1059:1065 the uncharacterized multi-pass transmembrane protein TMEM117 1014:1073 the uncharacterized multi-pass transmembrane protein TMEM117 1014:1073 This was supported by the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117, as confirmed experimentally. 27391701 2 47 theme cell 373:376 arg1 signaling 378:386 cell signaling 373:386 cell signaling 373:386 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 3 48 theme most 613:616 arg1 studies 618:624 most studies 613:624 most studies 613:624 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 2 49 theme containing 231:240 arg1 IDP 252:254 IDP 252:254 IDP 252:254 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 49 theme containing 231:240 arg1 Proteins 242:249 Intrinsic Disorder containing Proteins 212:249 Intrinsic Disorder containing Proteins (IDP) 212:255 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 10 50 from hubs 1713:1716 arg1 events 1737:1742 cellular signal events 1721:1742 cellular signal events 1721:1742 Altogether our analyses strongly support the notion that the transmembrane IDPs act as hubs in cellular signal events. 27391701 5 51 theme multi-pass 811:820 arg1 proteins 836:843 multi-pass transmembrane proteins 811:843 multi-pass transmembrane proteins 811:843 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 10 52 theme cellular 1721:1728 arg1 events 1737:1742 cellular signal events 1721:1742 cellular signal events 1721:1742 Altogether our analyses strongly support the notion that the transmembrane IDPs act as hubs in cellular signal events. 27391701 6 53 theme uncharacterized 1018:1032 arg1 protein 1059:1065 the uncharacterized multi-pass transmembrane protein TMEM117 1014:1073 the uncharacterized multi-pass transmembrane protein TMEM117 1014:1073 This was supported by the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117, as confirmed experimentally. 27391701 2 54 theme Disorder 222:229 arg1 IDP 252:254 IDP 252:254 IDP 252:254 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 54 theme Disorder 222:229 arg1 Proteins 242:249 Intrinsic Disorder containing Proteins 212:249 Intrinsic Disorder containing Proteins (IDP) 212:255 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 8 55 theme crucial 1338:1344 arg1 sites 1329:1333 phosphorylation sites 1313:1333 phosphorylation sites 1313:1333 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 8 55 theme crucial 1338:1344 arg1 modification 1365:1376 a crucial post translational modification 1336:1376 a crucial post translational modification in signal transduction 1336:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 3 56 theme available 462:470 arg1 databases 472:480 publicly available databases 453:480 publicly available databases 453:480 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 5 57 theme algorithms 935:944 arg1 confidence 901:910 the confidence 897:910 the confidence of topology prediction algorithms 897:944 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 8 58 theme interaction 1487:1497 arg1 events 1499:1504 more protein-protein interaction events 1466:1504 more protein-protein interaction events 1466:1504 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 2 59 theme Intrinsic 212:220 arg1 IDP 252:254 IDP 252:254 IDP 252:254 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 59 theme Intrinsic 212:220 arg1 Proteins 242:249 Intrinsic Disorder containing Proteins 212:249 Intrinsic Disorder containing Proteins (IDP) 212:255 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 3 60 dep prevalence 509:518 arg1 the 505:507 the 505:507 the 505:507 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 9 61 from regions 1571:1577 arg1 enriched 1537:1544 enriched 1537:1544 enriched 1537:1544 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 7 62 theme ion 1253:1255 arg1 binding 1257:1263 ion binding 1253:1263 ion binding 1253:1263 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 1 63 theme peculiar 134:141 arg1 stretches 143:151 peculiar stretches 134:151 peculiar stretches of amino acids that lack stable conformations in solution 134:209 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 1 63 theme peculiar 134:141 arg1 regions 115:121 Intrinsically disordered regions 90:121 Intrinsically disordered regions (IDRs) 90:128 Intrinsically disordered regions (IDRs) are peculiar stretches of amino acids that lack stable conformations in solution. 27391701 9 64 theme Recognition 1596:1606 arg1 Features 1608:1615 Molecular Recognition Features 1586:1615 Molecular Recognition Features (MoRFs) 1586:1623 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 9 64 theme Recognition 1596:1606 arg1 MoRFs 1618:1622 MoRFs 1618:1622 MoRFs 1618:1622 Accordingly, IDPs were highly enriched in short protein binding regions called Molecular Recognition Features (MoRFs). 27391701 2 65 theme IDR 307:309 arg1 presence 276:283 the presence 272:283 the presence of at least one large IDR 272:309 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 4 66 theme acids 705:709 arg1 IDR 689:691 at least one IDR 676:691 at least one IDR of 30 amino acids 676:709 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 4 67 theme amino 699:703 arg1 acids 705:709 30 amino acids 696:709 30 amino acids 696:709 We found that 50% of transmembrane proteins have at least one IDR of 30 amino acids or more. 27391701 2 68 theme large 301:305 arg1 IDR 307:309 at least one large IDR 288:309 at least one large IDR 288:309 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 8 69 from sites 1329:1333 arg1 enriched 1301:1308 enriched 1301:1308 enriched 1301:1308 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 7 70 theme cell 1224:1227 arg1 adhesion 1229:1236 cell adhesion 1224:1236 cell adhesion 1224:1236 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 3 71 from function 524:531 arg1 proteins 571:578 transmembrane proteins 557:578 transmembrane proteins 557:578 Here we used computational analyses and publicly available databases to deepen insight into the prevalence and function of IDRs specifically in transmembrane proteins, which are somewhat neglected in most studies. 27391701 0 72 dep Disorder 10:17 arg1 Roles 46:50 Roles 46:50 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 0:88 Intrinsic Disorder in Transmembrane Proteins: Roles in Signaling and Topology Prediction. 27391701 6 73 theme protein 1059:1065 arg1 topology 1002:1009 the topology 998:1009 the topology of the uncharacterized multi-pass transmembrane protein TMEM117 998:1073 This was supported by the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117, as confirmed experimentally. 27391701 2 74 theme multiple 335:342 arg1 signaling 378:386 cell signaling 373:386 cell signaling 373:386 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 74 theme multiple 335:342 arg1 processes 353:361 multiple cellular processes 335:361 multiple cellular processes including cell signaling, DNA binding and cancer 335:410 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 74 theme multiple 335:342 arg1 cancer 405:410 cancer 405:410 cancer 405:410 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 2 74 theme multiple 335:342 arg1 binding 393:399 DNA binding 389:399 DNA binding 389:399 Intrinsic Disorder containing Proteins (IDP) are defined by the presence of at least one large IDR and have been linked to multiple cellular processes including cell signaling, DNA binding and cancer. 27391701 6 75 theme topology 1002:1009 arg1 prediction 984:993 the successful prediction 969:993 the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117 969:1073 This was supported by the successful prediction of the topology of the uncharacterized multi-pass transmembrane protein TMEM117, as confirmed experimentally. 27391701 7 76 from enriched 1146:1153 arg1 cell 1158:1161 cell projection and axons 1158:1182 cell 1158:1161 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 7 76 from enriched 1146:1153 arg1 axons 1178:1182 axons 1178:1182 axons 1178:1182 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 27391701 5 77 theme disorder 862:869 arg1 prediction 871:880 disorder prediction 862:880 disorder prediction 862:880 Interestingly, these domains preferentially localize to the cytoplasmic side especially of multi-pass transmembrane proteins, suggesting that disorder prediction could increase the confidence of topology prediction algorithms. 27391701 8 78 from modification 1365:1376 arg1 transduction 1388:1399 signal transduction 1381:1399 signal transduction 1381:1399 In addition, we found that IDP are enriched in phosphorylation sites, a crucial post translational modification in signal transduction, when compared to fully ordered proteins and to be implicated in more protein-protein interaction events. 27391701 7 79 theme important 1206:1214 arg1 role 1216:1219 an important role 1203:1219 an important role 1203:1219 Pathway analysis indicated that IDPs are enriched in cell projection and axons and appear to play an important role in cell adhesion, signaling and ion binding. 9748270 10 0 theme keratinocytes 1539:1551 arg1 treatment 1519:1527 treatment 1519:1527 treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, 1519:1646 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 5 1 theme 120-kDa 670:676 arg1 polypeptides 678:689 three 120-kDa polypeptides 664:689 three 120-kDa polypeptides 664:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 8 2 theme 6-kb 1172:1175 arg1 mRNA 1177:1180 an identical 6-kb mRNA 1159:1180 an identical 6-kb mRNA 1159:1180 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 5 3 theme intracellular 574:586 arg1 domain 588:593 The globular intracellular domain 561:593 The globular intracellular domain 561:593 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 3 theme intracellular 574:586 arg1 disulfide-linked 599:614 disulfide-linked 599:614 disulfide-linked 599:614 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 10 4 theme furin 1583:1587 arg1 convertases 1600:1610 furin proprotein convertases 1583:1610 furin proprotein convertases 1583:1610 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 2 5 theme XVII 143:146 arg1 sequence 116:123 The cDNA sequence 107:123 The cDNA sequence of human collagen XVII 107:146 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 2 6 theme human 128:132 arg1 XVII 143:146 human collagen XVII 128:146 human collagen XVII 128:146 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 5 7 theme extracellular 640:652 arg1 domain 654:659 the N-glycosylated extracellular domain 621:659 the N-glycosylated extracellular domain of three 120-kDa polypeptides 621:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 7 theme extracellular 640:652 arg1 triple-helical 695:708 triple-helical 695:708 triple-helical 695:708 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 7 theme extracellular 640:652 arg1 polypeptides 678:689 three 120-kDa polypeptides 664:689 three 120-kDa polypeptides 664:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 9 8 theme alpha1 1462:1467 arg1 chain 1475:1479 the 180-kDa alpha1(XVII) chain 1450:1479 the 180-kDa alpha1(XVII) chain 1450:1479 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 12 9 theme triple-helical 2023:2036 arg1 ectodomain 2038:2047 a specifically processed, soluble triple-helical ectodomain 1989:2047 ectodomain 2038:2047 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 8 10 theme Northern 1039:1046 arg1 analysis 1053:1060 Northern blot analysis 1039:1060 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA 1039:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 10 11 theme decanoyl-RVKR-chloromethyl 1613:1638 arg1 ketone 1640:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 10 11 theme decanoyl-RVKR-chloromethyl 1613:1638 arg1 inhibitor 1570:1578 a synthetic inhibitor 1558:1578 a synthetic inhibitor of furin proprotein convertases 1558:1610 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 11 12 theme proteolytic 1862:1872 arg1 processing 1874:1883 furin-mediated proteolytic processing 1847:1883 furin-mediated proteolytic processing 1847:1883 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 10 13 theme convertases 1600:1610 arg1 ketone 1640:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 10 13 theme convertases 1600:1610 arg1 inhibitor 1570:1578 a synthetic inhibitor 1558:1578 a synthetic inhibitor of furin proprotein convertases 1558:1610 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 2 14 theme biochemical 205:215 arg1 characterization 217:232 a biochemical characterization 203:232 a biochemical characterization of this structure 203:250 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 3 15 theme domain-specific 289:303 arg1 antibodies 305:314 domain-specific antibodies 289:314 domain-specific antibodies against recombinant collagen XVII fragments 289:358 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 4 16 theme collagen 459:466 arg1 XVII 468:471 Full-length collagen XVII 447:471 Full-length collagen XVII 447:471 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 2 17 theme structure 242:250 arg1 characterization 217:232 a biochemical characterization 203:232 a biochemical characterization of this structure 203:250 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 7 18 theme polypeptides 1025:1036 arg1 molecule 999:1006 a triple-helical, N-glycosylated molecule 966:1006 molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides 912:1036 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 3 19 theme collagen 336:343 arg1 fragments 350:358 recombinant collagen XVII fragments 324:358 recombinant collagen XVII fragments 324:358 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 10 20 with treatment 1519:1527 arg1 ketone 1640:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 10 20 with treatment 1519:1527 arg1 inhibitor 1570:1578 a synthetic inhibitor 1558:1578 a synthetic inhibitor of furin proprotein convertases 1558:1610 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 4 21 theme transmembrane 500:512 arg1 molecule 514:521 a homotrimeric transmembrane molecule 485:521 a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains 485:558 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 7 22 dep triple-helical 968:981 arg1 N-glycosylated 984:997 N-glycosylated 984:997 N-glycosylated 984:997 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 11 23 theme collagen 1814:1821 arg1 ectodomain 1800:1809 a specifically cleaved ectodomain 1777:1809 a specifically cleaved ectodomain 1777:1809 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 11 23 theme collagen 1814:1821 arg1 collagen 1814:1821 collagen XVII 1814:1826 collagen XVII 1814:1826 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 1 24 theme full-length 47:57 arg1 protein 73:79 A full-length transmembrane protein 45:79 A full-length transmembrane protein 45:79 A full-length transmembrane protein and a soluble ectodomain. 9748270 0 25 from keratinocytes 30:42 arg1 forms 4:8 Two forms 0:8 Two forms of collagen XVII in keratinocytes 0:42 Two forms of collagen XVII in keratinocytes. 9748270 6 26 theme second 743:748 arg1 form 759:762 A second, soluble form 741:762 A second, soluble form of collagen XVII in keratinocyte culture media 741:809 A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. 9748270 3 27 theme epithelial 429:438 arg1 cells 440:444 epithelial cells 429:444 epithelial cells 429:444 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 12 28 theme transmembrane 1937:1949 arg1 XVII 1901:1904 collagen XVII 1892:1904 collagen XVII 1892:1904 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 12 28 theme transmembrane 1937:1949 arg1 collagen 1951:1958 an unusual type II transmembrane collagen 1918:1958 an unusual type II transmembrane collagen 1918:1958 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 5 29 theme globular 565:572 arg1 domain 588:593 The globular intracellular domain 561:593 The globular intracellular domain 561:593 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 29 theme globular 565:572 arg1 disulfide-linked 599:614 disulfide-linked 599:614 disulfide-linked 599:614 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 0 30 from XVII 22:25 arg1 keratinocytes 30:42 keratinocytes 30:42 keratinocytes 30:42 Two forms of collagen XVII in keratinocytes. 9748270 7 31 theme ectodomain 954:963 arg1 properties 922:931 molecular properties 912:931 molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides 912:1036 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 11 32 theme soluble 1738:1744 arg1 polypeptide 1754:1764 the soluble 120-kDa polypeptide 1734:1764 the soluble 120-kDa polypeptide 1734:1764 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 8 33 dep polypeptides 1225:1236 arg1 both 1199:1202 both 1199:1202 both 1199:1202 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 8 33 dep polypeptides 1225:1236 arg1 120-kDa 1217:1223 120-kDa 1217:1223 120-kDa 1217:1223 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 6 34 dep second 743:748 arg1 soluble 751:757 soluble 751:757 soluble 751:757 A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. 9748270 2 35 theme unusual 160:166 arg1 protein 190:196 an unusual type II transmembrane protein 157:196 an unusual type II transmembrane protein 157:196 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 0 36 theme XVII 22:25 arg1 forms 4:8 Two forms 0:8 Two forms of collagen XVII in keratinocytes 0:42 Two forms of collagen XVII in keratinocytes. 9748270 8 37 theme 3 1117:1117 arg1 end 1120:1122 either the distal 5'or the distal 3' end 1083:1122 either the distal 5'or the distal 3' end of the collagen XVII cDNA 1083:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 4 38 theme 180-kDa 532:538 arg1 chains 553:558 three 180-kDa alpha1(XVII) chains 526:558 three 180-kDa alpha1(XVII) chains 526:558 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 10 39 theme normal 1532:1537 arg1 keratinocytes 1539:1551 normal keratinocytes 1532:1551 normal keratinocytes 1532:1551 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 6 40 theme keratinocyte 784:795 arg1 media 805:809 keratinocyte culture media 784:809 keratinocyte culture media 784:809 A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. 9748270 12 41 theme type 1929:1932 arg1 XVII 1901:1904 collagen XVII 1892:1904 collagen XVII 1892:1904 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 12 41 theme type 1929:1932 arg1 collagen 1951:1958 an unusual type II transmembrane collagen 1918:1958 an unusual type II transmembrane collagen 1918:1958 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 9 42 theme homozygous 1381:1390 arg1 mutation 1401:1408 a homozygous nonsense mutation 1379:1408 a homozygous nonsense mutation in the COL17A1 gene 1379:1428 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 12 43 theme collagen 1892:1899 arg1 XVII 1901:1904 collagen XVII 1892:1904 collagen XVII 1892:1904 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 12 43 theme collagen 1892:1899 arg1 collagen 1951:1958 an unusual type II transmembrane collagen 1918:1958 an unusual type II transmembrane collagen 1918:1958 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 7 44 theme XVII 949:952 arg1 ectodomain 954:963 the collagen XVII ectodomain 936:963 the collagen XVII ectodomain 936:963 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 10 45 theme synthetic 1560:1568 arg1 ketone 1640:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 decanoyl-RVKR-chloromethyl ketone 1613:1645 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 10 45 theme synthetic 1560:1568 arg1 inhibitor 1570:1578 a synthetic inhibitor 1558:1578 a synthetic inhibitor of furin proprotein convertases 1558:1610 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 10 46 theme polypeptide 1688:1698 arg1 generation 1662:1671 the generation 1658:1671 the generation of the 120-kDa polypeptide 1658:1698 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 8 47 theme XVII 1140:1143 arg1 cDNA 1145:1148 the collagen XVII cDNA 1127:1148 the collagen XVII cDNA 1127:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 2 48 theme cDNA 111:114 arg1 sequence 116:123 The cDNA sequence 107:123 The cDNA sequence of human collagen XVII 107:146 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 10 49 theme proprotein 1589:1598 arg1 convertases 1600:1610 furin proprotein convertases 1583:1610 furin proprotein convertases 1583:1610 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 5 50 theme polypeptides 678:689 arg1 domain 654:659 the N-glycosylated extracellular domain 621:659 the N-glycosylated extracellular domain of three 120-kDa polypeptides 621:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 50 theme polypeptides 678:689 arg1 triple-helical 695:708 triple-helical 695:708 triple-helical 695:708 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 50 theme polypeptides 678:689 arg1 polypeptides 678:689 three 120-kDa polypeptides 664:689 three 120-kDa polypeptides 664:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 8 51 theme identical 1162:1170 arg1 mRNA 1177:1180 an identical 6-kb mRNA 1159:1180 an identical 6-kb mRNA 1159:1180 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 2 52 theme collagen 134:141 arg1 XVII 143:146 human collagen XVII 128:146 human collagen XVII 128:146 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 5 53 theme N-glycosylated 625:638 arg1 domain 654:659 the N-glycosylated extracellular domain 621:659 the N-glycosylated extracellular domain of three 120-kDa polypeptides 621:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 53 theme N-glycosylated 625:638 arg1 triple-helical 695:708 triple-helical 695:708 triple-helical 695:708 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 53 theme N-glycosylated 625:638 arg1 polypeptides 678:689 three 120-kDa polypeptides 664:689 three 120-kDa polypeptides 664:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 7 54 theme soluble 889:895 arg1 form 897:900 The soluble form 885:900 The soluble form 885:900 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 9 55 theme 180-kDa 1454:1460 arg1 XVII 1469:1472 XVII 1469:1472 XVII 1469:1472 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 9 55 theme 180-kDa 1454:1460 arg1 alpha1 1462:1467 180-kDa alpha1 1454:1467 the 180-kDa alpha1(XVII) chain 1450:1479 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 8 56 theme blot 1048:1051 arg1 analysis 1053:1060 Northern blot analysis 1039:1060 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA 1039:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 12 57 theme soluble 2015:2021 arg1 ectodomain 2038:2047 a specifically processed, soluble triple-helical ectodomain 1989:2047 ectodomain 2038:2047 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 7 58 dep properties 922:931 arg1 molecule 999:1006 a triple-helical, N-glycosylated molecule 966:1006 molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides 912:1036 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 4 59 theme Full-length 447:457 arg1 XVII 468:471 Full-length collagen XVII 447:471 Full-length collagen XVII 447:471 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 8 60 theme same 1263:1266 arg1 mRNA 1268:1271 the same mRNA 1259:1271 the same mRNA 1259:1271 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 11 61 theme furin-mediated 1847:1860 arg1 processing 1874:1883 furin-mediated proteolytic processing 1847:1883 furin-mediated proteolytic processing 1847:1883 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 9 62 from mutation 1401:1408 arg1 gene 1425:1428 the COL17A1 gene 1413:1428 the COL17A1 gene 1413:1428 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 9 63 theme 120-kDa 1489:1495 arg1 polypeptide 1497:1507 the 120-kDa polypeptide 1485:1507 the 120-kDa polypeptide 1485:1507 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 12 64 theme processed 2004:2012 arg1 ectodomain 2038:2047 a specifically processed, soluble triple-helical ectodomain 1989:2047 ectodomain 2038:2047 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 5 65 theme physiological 713:725 arg1 temperatures 727:738 physiological temperatures 713:738 physiological temperatures 713:738 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 8 66 theme 120-kDa 1287:1293 arg1 polypeptide 1295:1305 the 120-kDa polypeptide 1283:1305 the 120-kDa polypeptide 1283:1305 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 3 67 theme XVII 345:348 arg1 fragments 350:358 recombinant collagen XVII fragments 324:358 recombinant collagen XVII fragments 324:358 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 11 68 theme cleaved 1792:1798 arg1 ectodomain 1800:1809 a specifically cleaved ectodomain 1777:1809 a specifically cleaved ectodomain 1777:1809 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 11 68 theme cleaved 1792:1798 arg1 collagen 1814:1821 collagen XVII 1814:1826 collagen XVII 1814:1826 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 12 69 theme first 1969:1973 arg1 collagen 1975:1982 the first collagen 1965:1982 the first collagen with a specifically processed, soluble triple-helical ectodomain 1965:2047 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 7 70 theme 120-kDa 1017:1023 arg1 polypeptides 1025:1036 three 120-kDa polypeptides 1011:1036 three 120-kDa polypeptides 1011:1036 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 3 71 theme recombinant 324:334 arg1 fragments 350:358 recombinant collagen XVII fragments 324:358 recombinant collagen XVII fragments 324:358 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 0 72 from forms 4:8 arg1 keratinocytes 30:42 keratinocytes 30:42 keratinocytes 30:42 Two forms of collagen XVII in keratinocytes. 9748270 11 73 theme 120-kDa 1746:1752 arg1 polypeptide 1754:1764 the soluble 120-kDa polypeptide 1734:1764 the soluble 120-kDa polypeptide 1734:1764 These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. 9748270 8 74 with analysis 1053:1060 arg1 probes 1067:1072 probes 1067:1072 probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA 1067:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 8 75 theme 5 1101:1101 arg1 end 1120:1122 either the distal 5'or the distal 3' end 1083:1122 either the distal 5'or the distal 3' end of the collagen XVII cDNA 1083:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 5 76 gly N-glycosylated 625:638 arg1 domain 654:659 the N-glycosylated extracellular domain 621:659 the N-glycosylated extracellular domain of three 120-kDa polypeptides 621:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 76 gly N-glycosylated 625:638 arg1 triple-helical 695:708 triple-helical 695:708 triple-helical 695:708 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 76 gly N-glycosylated 625:638 arg1 polypeptides 678:689 three 120-kDa polypeptides 664:689 three 120-kDa polypeptides 664:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 1 77 theme transmembrane 59:71 arg1 protein 73:79 A full-length transmembrane protein 45:79 A full-length transmembrane protein 45:79 A full-length transmembrane protein and a soluble ectodomain. 9748270 4 78 theme homotrimeric 487:498 arg1 molecule 514:521 a homotrimeric transmembrane molecule 485:521 a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains 485:558 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 10 79 theme 120-kDa 1680:1686 arg1 polypeptide 1688:1698 the 120-kDa polypeptide 1676:1698 the 120-kDa polypeptide 1676:1698 Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. 9748270 7 80 theme triple-helical 968:981 arg1 molecule 999:1006 a triple-helical, N-glycosylated molecule 966:1006 molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides 912:1036 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 4 81 theme chains 553:558 arg1 molecule 514:521 a homotrimeric transmembrane molecule 485:521 a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains 485:558 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 0 82 theme collagen 13:20 arg1 XVII 22:25 collagen XVII 13:25 collagen XVII in keratinocytes 13:42 Two forms of collagen XVII in keratinocytes. 9748270 12 83 with collagen 1975:1982 arg1 ectodomain 2038:2047 a specifically processed, soluble triple-helical ectodomain 1989:2047 ectodomain 2038:2047 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 12 84 theme unusual 1921:1927 arg1 XVII 1901:1904 collagen XVII 1892:1904 collagen XVII 1892:1904 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 12 84 theme unusual 1921:1927 arg1 collagen 1951:1958 an unusual type II transmembrane collagen 1918:1958 an unusual type II transmembrane collagen 1918:1958 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 1 85 theme soluble 87:93 arg1 ectodomain 95:104 a soluble ectodomain 85:104 a soluble ectodomain 85:104 A full-length transmembrane protein and a soluble ectodomain. 9748270 3 86 theme molecular 379:387 arg1 forms 389:393 two molecular forms 375:393 two molecular forms of the collagen 375:409 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 2 87 theme type 168:171 arg1 protein 190:196 an unusual type II transmembrane protein 157:196 an unusual type II transmembrane protein 157:196 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 12 88 theme II 1934:1935 arg1 XVII 1901:1904 collagen XVII 1892:1904 collagen XVII 1892:1904 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 12 88 theme II 1934:1935 arg1 collagen 1951:1958 an unusual type II transmembrane collagen 1918:1958 an unusual type II transmembrane collagen 1918:1958 Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. 9748270 3 89 theme human 414:418 arg1 skin 420:423 human skin 414:423 human skin 414:423 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 4 90 theme alpha1 540:545 arg1 chains 553:558 three 180-kDa alpha1(XVII) chains 526:558 three 180-kDa alpha1(XVII) chains 526:558 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 6 91 theme culture 797:803 arg1 media 805:809 keratinocyte culture media 784:809 keratinocyte culture media 784:809 A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. 9748270 3 92 theme collagen 402:409 arg1 forms 389:393 two molecular forms 375:393 two molecular forms of the collagen 375:409 Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. 9748270 4 93 theme XVII 547:550 arg1 chains 553:558 three 180-kDa alpha1(XVII) chains 526:558 three 180-kDa alpha1(XVII) chains 526:558 Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. 9748270 8 94 dep both 1199:1202 arg1 180- 1208:1211 the 180- 1204:1211 the 180- 1204:1211 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 7 95 theme collagen 940:947 arg1 ectodomain 954:963 the collagen XVII ectodomain 936:963 the collagen XVII ectodomain 936:963 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 9 96 theme nonsense 1392:1399 arg1 mutation 1401:1408 a homozygous nonsense mutation 1379:1408 a homozygous nonsense mutation in the COL17A1 gene 1379:1428 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 9748270 8 97 theme collagen 1131:1138 arg1 cDNA 1145:1148 the collagen XVII cDNA 1127:1148 the collagen XVII cDNA 1127:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 2 98 theme transmembrane 176:188 arg1 protein 190:196 an unusual type II transmembrane protein 157:196 an unusual type II transmembrane protein 157:196 The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. 9748270 6 99 theme collagen 767:774 arg1 form 759:762 A second, soluble form 741:762 A second, soluble form of collagen XVII in keratinocyte culture media 741:809 A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. 9748270 5 100 from temperatures 727:738 arg1 domain 654:659 the N-glycosylated extracellular domain 621:659 the N-glycosylated extracellular domain of three 120-kDa polypeptides 621:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 100 from temperatures 727:738 arg1 triple-helical 695:708 triple-helical 695:708 triple-helical 695:708 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 5 100 from temperatures 727:738 arg1 polypeptides 678:689 three 120-kDa polypeptides 664:689 three 120-kDa polypeptides 664:689 The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. 9748270 8 101 theme cDNA 1145:1148 arg1 end 1120:1122 either the distal 5'or the distal 3' end 1083:1122 either the distal 5'or the distal 3' end of the collagen XVII cDNA 1083:1148 Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. 9748270 6 102 from form 759:762 arg1 media 805:809 keratinocyte culture media 784:809 keratinocyte culture media 784:809 A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. 9748270 7 103 theme molecular 912:920 arg1 properties 922:931 molecular properties 912:931 molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides 912:1036 The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120-kDa polypeptides. 9748270 9 104 theme COL17A1 1417:1423 arg1 gene 1425:1428 the COL17A1 gene 1413:1428 the COL17A1 gene 1413:1428 In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. 11953431 3 0 theme gamma-glutamyl 359:372 arg1 hydrolase 374:382 human gamma-glutamyl hydrolase 353:382 human gamma-glutamyl hydrolase 353:382 The crystal structure of human gamma-glutamyl hydrolase, determined at 1.6-A resolution, reveals that the protein is a homodimer. 11953431 4 1 from alpha-helices 650:662 arg1 side 672:675 each side 667:675 each side 667:675 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 5 2 theme class 722:726 arg1 amidotransferase 740:755 the class I glutamine amidotransferase 718:755 the class I glutamine amidotransferase domains 718:763 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 10 3 theme intermediate 1507:1518 arg1 complex 1472:1478 the corresponding complex 1454:1478 the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase 1454:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 10 4 theme carbamoyl-phosphate 1540:1558 arg1 synthetase 1560:1569 Escherichia coli carbamoyl-phosphate synthetase 1523:1569 Escherichia coli carbamoyl-phosphate synthetase 1523:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 3 5 theme hydrolase 374:382 arg1 structure 340:348 The crystal structure 328:348 The crystal structure 328:348 The crystal structure of human gamma-glutamyl hydrolase, determined at 1.6-A resolution, reveals that the protein is a homodimer. 11953431 5 6 from extensions 812:821 arg1 loops 831:835 four loops 826:835 four loops 826:835 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 9 7 theme Several 1276:1282 arg1 Lys-223 1345:1351 Lys-223 1345:1351 Lys-223 1345:1351 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 7 theme Several 1276:1282 arg1 His-171 1323:1329 His-171 1323:1329 His-171 1323:1329 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 7 theme Several 1276:1282 arg1 residues 1294:1301 Several conserved residues 1276:1301 Several conserved residues 1276:1301 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 7 theme Several 1276:1282 arg1 Glu-114 1314:1320 Glu-114 1314:1320 Glu-114 1314:1320 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 7 theme Several 1276:1282 arg1 Gln-218 1332:1338 Gln-218 1332:1338 Gln-218 1332:1338 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 10 8 theme intermediate 1431:1442 arg1 Modeling 1394:1401 Modeling 1394:1401 Modeling 1394:1401 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 10 9 theme glutamate 1487:1495 arg1 intermediate 1507:1518 the glutamate thioester intermediate 1483:1518 the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase 1483:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 4 10 theme gamma-glutamyl 489:502 arg1 hydrolase 504:512 human gamma-glutamyl hydrolase 483:512 human gamma-glutamyl hydrolase 483:512 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 7 11 theme human 1008:1012 arg1 hydrolase 1029:1037 human gamma-glutamyl hydrolase 1008:1037 human gamma-glutamyl hydrolase 1008:1037 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 8 12 from end 1249:1251 arg1 cleft 1221:1225 a large l-shaped cleft 1204:1225 a large l-shaped cleft that is closed at one end and open at the other 1204:1273 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 8 12 from end 1249:1251 arg1 closed 1235:1240 closed 1235:1240 closed 1235:1240 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 7 13 theme I 1053:1053 arg1 amidotransferase 1065:1080 the class I glutamine amidotransferase 1043:1080 the class I glutamine amidotransferase family 1043:1087 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 1 14 theme class 66:70 arg1 amidotransferase 84:99 A class I glatamine amidotransferase 64:99 A class I glatamine amidotransferase 64:99 A class I glatamine amidotransferase adapted for a complex substate. 11953431 4 15 theme three 635:639 arg1 alpha-helices 650:662 three and five alpha-helices 635:662 alpha-helices 650:662 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 8 16 theme large 1206:1210 arg1 cleft 1221:1225 a large l-shaped cleft 1204:1225 a large l-shaped cleft that is closed at one end and open at the other 1204:1273 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 8 16 theme large 1206:1210 arg1 closed 1235:1240 closed 1235:1240 closed 1235:1240 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 6 17 theme binding 918:924 arg1 cleft 926:930 the substrate binding cleft 904:930 the substrate binding cleft 904:930 These insertions are important for defining the substrate binding cleft and/or the dimer interface. 11953431 2 18 dep folyl 275:279 arg1 metabolism 316:325 poly-gamma-glutamate metabolism 295:325 poly-gamma-glutamate metabolism 295:325 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 3 19 theme 1.6-A 399:403 arg1 resolution 405:414 1.6-A resolution 399:414 1.6-A resolution 399:414 The crystal structure of human gamma-glutamyl hydrolase, determined at 1.6-A resolution, reveals that the protein is a homodimer. 11953431 6 20 theme substrate 908:916 arg1 cleft 926:930 the substrate binding cleft 904:930 the substrate binding cleft 904:930 These insertions are important for defining the substrate binding cleft and/or the dimer interface. 11953431 7 21 theme sequence 964:971 arg1 motifs 973:978 Two sequence motifs 960:978 Two sequence motifs 960:978 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 1 22 theme glatamine 74:82 arg1 amidotransferase 84:99 A class I glatamine amidotransferase 64:99 A class I glatamine amidotransferase 64:99 A class I glatamine amidotransferase adapted for a complex substate. 11953431 2 23 from enzyme 265:270 arg1 folyl 275:279 folyl 275:279 folyl 275:279 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 2 23 from enzyme 265:270 arg1 antifolyl 285:293 antifolyl 285:293 antifolyl 285:293 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 8 24 theme cleft 1221:1225 arg1 center 1194:1199 the center 1190:1199 the center of a large l-shaped cleft that is closed at one end and open at the other 1190:1273 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 10 25 theme thioester 1421:1429 arg1 intermediate 1431:1442 a methotrexate thioester intermediate 1406:1442 a methotrexate thioester intermediate 1406:1442 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 10 26 theme corresponding 1458:1470 arg1 complex 1472:1478 the corresponding complex 1454:1478 the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase 1454:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 10 27 theme methotrexate 1408:1419 arg1 intermediate 1431:1442 a methotrexate thioester intermediate 1406:1442 a methotrexate thioester intermediate 1406:1442 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 0 28 theme Three-dimensional 0:16 arg1 structure 18:26 Three-dimensional structure 0:26 Three-dimensional structure of human gamma -glutamyl hydrolase 0:61 Three-dimensional structure of human gamma -glutamyl hydrolase. 11953431 10 29 theme pteroyl 1634:1640 arg1 group 1642:1646 the pteroyl group 1630:1646 the pteroyl group 1630:1646 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 9 30 theme conserved 1284:1292 arg1 Lys-223 1345:1351 Lys-223 1345:1351 Lys-223 1345:1351 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 30 theme conserved 1284:1292 arg1 His-171 1323:1329 His-171 1323:1329 His-171 1323:1329 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 30 theme conserved 1284:1292 arg1 residues 1294:1301 Several conserved residues 1276:1301 Several conserved residues 1276:1301 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 30 theme conserved 1284:1292 arg1 Glu-114 1314:1320 Glu-114 1314:1320 Glu-114 1314:1320 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 9 30 theme conserved 1284:1292 arg1 Gln-218 1332:1338 Gln-218 1332:1338 Gln-218 1332:1338 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 4 31 theme human 483:487 arg1 hydrolase 504:512 human gamma-glutamyl hydrolase 483:512 human gamma-glutamyl hydrolase 483:512 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 0 32 theme human 31:35 arg1 hydrolase 53:61 human gamma -glutamyl hydrolase 31:61 human gamma -glutamyl hydrolase 31:61 Three-dimensional structure of human gamma -glutamyl hydrolase. 11953431 5 33 theme I 728:728 arg1 amidotransferase 740:755 the class I glutamine amidotransferase 718:755 the class I glutamine amidotransferase domains 718:763 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 10 34 with orientation 1613:1623 arg1 group 1642:1646 the pteroyl group 1630:1646 the pteroyl group 1630:1646 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 10 35 dep Escherichia 1523:1533 arg1 coli 1535:1538 coli 1535:1538 coli 1535:1538 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 10 36 theme cleft 1675:1679 arg1 end 1664:1666 the open end 1655:1666 the open end of the cleft 1655:1679 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 5 37 theme C 848:848 arg1 terminus 850:857 the C terminus 844:857 the C terminus 844:857 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 5 38 theme glutamine 730:738 arg1 amidotransferase 740:755 the class I glutamine amidotransferase 718:755 the class I glutamine amidotransferase domains 718:763 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 2 39 theme chain 203:207 arg1 cleavage 172:179 the cleavage 168:179 the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates 168:246 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 7 40 theme amidotransferase 1065:1080 arg1 family 1082:1087 the class I glutamine amidotransferase family 1043:1087 the class I glutamine amidotransferase family 1043:1087 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 6 41 theme dimer 943:947 arg1 interface 949:957 the dimer interface 939:957 the dimer interface 939:957 These insertions are important for defining the substrate binding cleft and/or the dimer interface. 11953431 4 42 contain contains 514:521 arg1 structure 470:478 The overall structure 458:478 The overall structure of human gamma-glutamyl hydrolase 458:512 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 4 42 contain contains 514:521 arg2 beta-strands 547:558 14 beta-strands 544:558 14 beta-strands 544:558 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 4 42 contain contains 514:521 arg2 alpha-helices 526:538 11 alpha-helices 523:538 11 alpha-helices 523:538 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 0 43 theme -glutamyl 43:51 arg1 hydrolase 53:61 human gamma -glutamyl hydrolase 31:61 human gamma -glutamyl hydrolase 31:61 Three-dimensional structure of human gamma -glutamyl hydrolase. 11953431 5 44 theme amidotransferase 740:755 arg1 domains 757:763 the class I glutamine amidotransferase domains 718:763 the class I glutamine amidotransferase domains 718:763 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 2 45 theme gamma-glutamyl 188:201 arg1 chain 203:207 the gamma-glutamyl chain 184:207 the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates 184:246 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 10 46 theme open 1659:1662 arg1 end 1664:1666 the open end 1655:1666 the open end of the cleft 1655:1679 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 0 47 theme gamma 37:41 arg1 hydrolase 53:61 human gamma -glutamyl hydrolase 31:61 human gamma -glutamyl hydrolase 31:61 Three-dimensional structure of human gamma -glutamyl hydrolase. 11953431 8 48 from located 1179:1185 arg1 center 1194:1199 the center 1190:1199 the center of a large l-shaped cleft that is closed at one end and open at the other 1190:1273 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 2 49 theme central 257:263 arg1 enzyme 265:270 a central enzyme 255:270 a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism 255:325 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 7 50 theme class 1047:1051 arg1 amidotransferase 1065:1080 the class I glutamine amidotransferase 1043:1080 the class I glutamine amidotransferase family 1043:1087 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 4 51 from sandwiched 621:630 arg1 fold 568:571 a fold 566:571 a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side 566:675 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 7 52 located found 984:988 arg2 motifs 973:978 Two sequence motifs 960:978 Two sequence motifs 960:978 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 7 52 located found 984:988 arg1 common 993:998 common 993:998 common 993:998 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 4 53 from fold 568:571 arg1 sandwiched 621:630 sandwiched 621:630 sandwiched 621:630 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 4 53 from fold 568:571 arg1 beta-sheet 607:616 beta-sheet 607:616 beta-sheet 607:616 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 4 54 theme hydrolase 504:512 arg1 structure 470:478 The overall structure 458:478 The overall structure of human gamma-glutamyl hydrolase 458:512 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 10 55 theme synthetase 1560:1569 arg1 intermediate 1507:1518 the glutamate thioester intermediate 1483:1518 the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase 1483:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 7 56 theme glutamine 1055:1063 arg1 amidotransferase 1065:1080 the class I glutamine amidotransferase 1043:1080 the class I glutamine amidotransferase family 1043:1087 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 1 57 theme complex 115:121 arg1 substate 123:130 a complex substate 113:130 a complex substate 113:130 A class I glatamine amidotransferase adapted for a complex substate. 11953431 0 58 theme hydrolase 53:61 arg1 structure 18:26 Three-dimensional structure 0:26 Three-dimensional structure of human gamma -glutamyl hydrolase 0:61 Three-dimensional structure of human gamma -glutamyl hydrolase. 11953431 3 59 theme human 353:357 arg1 hydrolase 374:382 human gamma-glutamyl hydrolase 353:382 human gamma-glutamyl hydrolase 353:382 The crystal structure of human gamma-glutamyl hydrolase, determined at 1.6-A resolution, reveals that the protein is a homodimer. 11953431 4 60 theme overall 462:468 arg1 structure 470:478 The overall structure 458:478 The overall structure of human gamma-glutamyl hydrolase 458:512 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 3 61 theme crystal 332:338 arg1 structure 340:348 The crystal structure 328:348 The crystal structure 328:348 The crystal structure of human gamma-glutamyl hydrolase, determined at 1.6-A resolution, reveals that the protein is a homodimer. 11953431 2 62 theme substrates 237:246 arg1 chain 203:207 the gamma-glutamyl chain 184:207 the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates 184:246 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 8 63 from other 1269:1273 arg1 open 1257:1260 open 1257:1260 open 1257:1260 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 9 64 theme substrate 1375:1383 arg1 binding 1385:1391 substrate binding 1375:1391 substrate binding 1375:1391 Several conserved residues, including Glu-114, His-171, Gln-218, and Lys-223, may be important for substrate binding. 11953431 2 65 theme poly-gamma-glutamate 295:314 arg1 metabolism 316:325 poly-gamma-glutamate metabolism 295:325 poly-gamma-glutamate metabolism 295:325 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 2 66 theme folylpoly-gamma-glutamyl 212:235 arg1 substrates 237:246 folylpoly-gamma-glutamyl substrates 212:246 folylpoly-gamma-glutamyl substrates 212:246 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 11953431 8 67 theme l-shaped 1212:1219 arg1 cleft 1221:1225 a large l-shaped cleft 1204:1225 a large l-shaped cleft that is closed at one end and open at the other 1204:1273 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 8 67 theme l-shaped 1212:1219 arg1 closed 1235:1240 closed 1235:1240 closed 1235:1240 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 7 68 theme gamma-glutamyl 1014:1027 arg1 hydrolase 1029:1037 human gamma-glutamyl hydrolase 1008:1037 human gamma-glutamyl hydrolase 1008:1037 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 8 69 located located 1179:1185 arg2 residues 1166:1173 These residues 1160:1173 These residues 1160:1173 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 8 69 located located 1179:1185 arg1 center 1194:1199 the center 1190:1199 the center of a large l-shaped cleft that is closed at one end and open at the other 1190:1273 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 4 70 theme five 645:648 arg1 alpha-helices 650:662 three and five alpha-helices 635:662 alpha-helices 650:662 The overall structure of human gamma-glutamyl hydrolase contains 11 alpha-helices and 14 beta-strands, with a fold in which a central eight-stranded beta-sheet is sandwiched by three and five alpha-helices on each side. 11953431 5 71 theme major 780:784 arg1 differences 786:796 the only major differences 771:796 the only major differences consisting of extensions in four loops and at the C terminus 771:857 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 8 72 from center 1194:1199 arg1 located 1179:1185 located 1179:1185 located 1179:1185 These residues are located in the center of a large l-shaped cleft that is closed at one end and open at the other. 11953431 10 73 theme Escherichia 1523:1533 arg1 synthetase 1560:1569 Escherichia coli carbamoyl-phosphate synthetase 1523:1569 Escherichia coli carbamoyl-phosphate synthetase 1523:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 5 74 with similar 699:705 arg1 differences 786:796 the only major differences 771:796 the only major differences consisting of extensions in four loops and at the C terminus 771:857 The topology is very similar to that of the class I glutamine amidotransferase domains, with the only major differences consisting of extensions in four loops and at the C terminus. 11953431 10 75 theme thioester 1497:1505 arg1 intermediate 1507:1518 the glutamate thioester intermediate 1483:1518 the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase 1483:1569 Modeling of a methotrexate thioester intermediate, based on the corresponding complex of the glutamate thioester intermediate of Escherichia coli carbamoyl-phosphate synthetase, indicates that the substrate binds in an orientation with the pteroyl group toward the open end of the cleft. 11953431 7 76 theme essential 1119:1127 arg1 Cys-110 1139:1145 Cys-110 1139:1145 Cys-110 1139:1145 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 7 76 theme essential 1119:1127 arg1 residues 1129:1136 the catalytically essential residues 1101:1136 the catalytically essential residues 1101:1136 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 7 76 theme essential 1119:1127 arg1 His-220 1151:1157 His-220 1151:1157 His-220 1151:1157 Two sequence motifs are found in common between human gamma-glutamyl hydrolase and the class I glutamine amidotransferase family and include the catalytically essential residues, Cys-110 and His-220. 11953431 2 77 theme gamma-Glutamyl 133:146 arg1 hydrolase 148:156 gamma-Glutamyl hydrolase 133:156 gamma-Glutamyl hydrolase 133:156 gamma-Glutamyl hydrolase catalyzes the cleavage of the gamma-glutamyl chain of folylpoly-gamma-glutamyl substrates and is a central enzyme in folyl and antifolyl poly-gamma-glutamate metabolism. 15014436 9 0 from Mutations 1060:1068 arg1 repeats 1080:1086 the T3 repeats 1073:1086 the T3 repeats of TSP-5/COMP 1073:1100 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 4 1 theme novel 619:623 arg1 arrangement 625:635 a novel arrangement 617:635 a novel arrangement 617:635 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 9 2 theme T3 1077:1078 arg1 repeats 1080:1086 the T3 repeats 1073:1086 the T3 repeats of TSP-5/COMP 1073:1100 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 1 3 theme extracellular 129:141 arg1 Thrombospondins 102:116 Thrombospondins 102:116 Thrombospondins (TSPs) 102:123 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 1 3 theme extracellular 129:141 arg1 regulators 143:152 extracellular regulators 129:152 extracellular regulators of cell-matrix interactions and cell phenotype 129:199 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 8 4 theme region 1052:1057 arg1 functions 1020:1028 the functions 1016:1028 the functions of the TSP C-terminal region 1016:1057 The central architectural role of calcium explains how it is critical for the functions of the TSP C-terminal region. 15014436 5 5 contain contains 684:691 arg2 sites 733:737 four strictly conserved calcium-binding sites 693:737 four strictly conserved calcium-binding sites 693:737 The CTD forms a lectin-like beta-sandwich and contains four strictly conserved calcium-binding sites. 15014436 5 5 contain contains 684:691 arg1 CTD 642:644 The CTD 638:644 The CTD 638:644 The CTD forms a lectin-like beta-sandwich and contains four strictly conserved calcium-binding sites. 15014436 6 6 theme hairpin 758:764 arg1 structure 766:774 the hairpin structure 754:774 the hairpin structure of T3 repeats 6 and 7 754:796 Disruption of the hairpin structure of T3 repeats 6 and 7 decreases protein secretion and stability. 15014436 8 7 theme architectural 954:966 arg1 role 968:971 The central architectural role 942:971 The central architectural role of calcium 942:982 The central architectural role of calcium explains how it is critical for the functions of the TSP C-terminal region. 15014436 8 8 theme central 946:952 arg1 role 968:971 The central architectural role 942:971 The central architectural role of calcium 942:982 The central architectural role of calcium explains how it is critical for the functions of the TSP C-terminal region. 15014436 0 9 theme type 86:89 arg1 repeats 93:99 the type 3 repeats 82:99 the type 3 repeats 82:99 Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. 15014436 7 10 theme T3 897:898 arg1 repeat 900:905 T3 repeat 7 897:907 T3 repeat 7 897:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 9 11 theme T3-CTD 1200:1205 arg1 assembly 1207:1214 the T3-CTD assembly 1196:1214 the T3-CTD assembly 1196:1214 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 2 12 theme type 271:274 arg1 region 228:233 The most highly conserved region 202:233 The most highly conserved region of all TSPs 202:245 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 2 12 theme type 271:274 arg1 repeats 283:289 the calcium-binding type 3 (T3) repeats 251:289 the calcium-binding type 3 (T3) repeats 251:289 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 7 13 theme RGD 884:886 arg1 motif 888:892 an RGD motif 881:892 an RGD motif in T3 repeat 7 881:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 3 14 theme compact 438:444 arg1 assembly 446:453 a compact assembly 436:453 a compact assembly 436:453 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, reveals a compact assembly. 15014436 5 15 theme lectin-like 654:664 arg1 beta-sandwich 666:678 a lectin-like beta-sandwich 652:678 a lectin-like beta-sandwich 652:678 The CTD forms a lectin-like beta-sandwich and contains four strictly conserved calcium-binding sites. 15014436 9 16 theme assembly 1207:1214 arg1 structure 1183:1191 the tertiary structure 1170:1191 the tertiary structure of the T3-CTD assembly 1170:1214 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 8 17 theme calcium 976:982 arg1 role 968:971 The central architectural role 942:971 The central architectural role of calcium 942:982 The central architectural role of calcium explains how it is critical for the functions of the TSP C-terminal region. 15014436 4 18 theme DxDxDGxxDxxD 549:560 arg1 motifs 562:567 two DxDxDGxxDxxD motifs 545:567 two DxDxDGxxDxxD motifs per repeat each 545:583 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 2 19 theme calcium-binding 255:269 arg1 region 228:233 The most highly conserved region 202:233 The most highly conserved region of all TSPs 202:245 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 2 19 theme calcium-binding 255:269 arg1 repeats 283:289 the calcium-binding type 3 (T3) repeats 251:289 the calcium-binding type 3 (T3) repeats 251:289 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 1 20 theme interactions 169:180 arg1 Thrombospondins 102:116 Thrombospondins 102:116 Thrombospondins (TSPs) 102:123 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 1 20 theme interactions 169:180 arg1 regulators 143:152 extracellular regulators 129:152 extracellular regulators of cell-matrix interactions and cell phenotype 129:199 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 5 21 theme calcium-binding 717:731 arg1 sites 733:737 four strictly conserved calcium-binding sites 693:737 four strictly conserved calcium-binding sites 693:737 The CTD forms a lectin-like beta-sandwich and contains four strictly conserved calcium-binding sites. 15014436 3 22 theme cell-binding 360:371 arg1 fragment 379:386 a cell-binding TSP-1 fragment 358:386 a cell-binding TSP-1 fragment 358:386 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, reveals a compact assembly. 15014436 1 23 theme cell-matrix 157:167 arg1 interactions 169:180 cell-matrix interactions 157:180 cell-matrix interactions 157:180 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 3 24 theme crystal 337:343 arg1 structure 345:353 The crystal structure 333:353 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, 333:426 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, reveals a compact assembly. 15014436 6 25 theme protein 808:814 arg1 secretion 816:824 protein secretion 808:824 protein secretion 808:824 Disruption of the hairpin structure of T3 repeats 6 and 7 decreases protein secretion and stability. 15014436 8 26 theme C-terminal 1041:1050 arg1 region 1052:1057 the TSP C-terminal region 1033:1057 the TSP C-terminal region 1033:1057 The central architectural role of calcium explains how it is critical for the functions of the TSP C-terminal region. 15014436 7 27 from attachment 867:876 arg1 repeat 900:905 T3 repeat 7 897:907 T3 repeat 7 897:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 3 28 theme fragment 379:386 arg1 structure 345:353 The crystal structure 333:353 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, 333:426 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, reveals a compact assembly. 15014436 7 29 theme cell 862:865 arg1 attachment 867:876 cell attachment 862:876 cell attachment of an RGD motif in T3 repeat 7 862:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 0 30 theme thrombospondin 15:28 arg1 fragment 41:48 a thrombospondin C-terminal fragment 13:48 a thrombospondin C-terminal fragment 13:48 Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. 15014436 7 31 theme motif 888:892 arg1 attachment 867:876 cell attachment 862:876 cell attachment of an RGD motif in T3 repeat 7 862:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 1 32 theme cell 186:189 arg1 phenotype 191:199 cell phenotype 186:199 cell phenotype 186:199 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 9 33 theme skeletal 1125:1132 arg1 disorders 1134:1142 two human skeletal disorders 1115:1142 two human skeletal disorders 1115:1142 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 7 34 from repeat 900:905 arg1 attachment 867:876 cell attachment 862:876 cell attachment of an RGD motif in T3 repeat 7 862:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 1 35 theme phenotype 191:199 arg1 Thrombospondins 102:116 Thrombospondins 102:116 Thrombospondins (TSPs) 102:123 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 1 35 theme phenotype 191:199 arg1 regulators 143:152 extracellular regulators 129:152 extracellular regulators of cell-matrix interactions and cell phenotype 129:199 Thrombospondins (TSPs) are extracellular regulators of cell-matrix interactions and cell phenotype. 15014436 0 36 theme fragment 41:48 arg1 Structure 0:8 Structure 0:8 Structure of a thrombospondin C-terminal fragment 0:48 Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. 15014436 4 37 theme calcium 601:607 arg1 ions 609:612 two calcium ions 597:612 two calcium ions 597:612 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 3 38 theme T3 404:405 arg1 repeats 407:413 three T3 repeats 398:413 three T3 repeats 398:413 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, reveals a compact assembly. 15014436 0 39 theme C-terminal 30:39 arg1 fragment 41:48 a thrombospondin C-terminal fragment 13:48 a thrombospondin C-terminal fragment 13:48 Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. 15014436 3 40 theme TSP-1 373:377 arg1 fragment 379:386 a cell-binding TSP-1 fragment 358:386 a cell-binding TSP-1 fragment 358:386 The crystal structure of a cell-binding TSP-1 fragment, spanning three T3 repeats and the CTD, reveals a compact assembly. 15014436 2 41 theme conserved 218:226 arg1 region 228:233 The most highly conserved region 202:233 The most highly conserved region of all TSPs 202:245 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 2 41 theme conserved 218:226 arg1 repeats 283:289 the calcium-binding type 3 (T3) repeats 251:289 the calcium-binding type 3 (T3) repeats 251:289 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 4 42 theme T3 460:461 arg1 repeats 463:469 The T3 repeats 456:469 The T3 repeats 456:469 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 9 43 theme TSP-5/COMP 1091:1100 arg1 repeats 1080:1086 the T3 repeats 1073:1086 the T3 repeats of TSP-5/COMP 1073:1100 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 8 44 theme TSP 1037:1039 arg1 region 1052:1057 the TSP C-terminal region 1033:1057 the TSP C-terminal region 1033:1057 The central architectural role of calcium explains how it is critical for the functions of the TSP C-terminal region. 15014436 4 45 dep motifs 562:567 arg1 each 580:583 each 580:583 each 580:583 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 2 46 theme globular 310:317 arg1 CTD 327:329 CTD 327:329 CTD 327:329 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 2 46 theme globular 310:317 arg1 domain 319:324 the C-terminal globular domain 295:324 the C-terminal globular domain (CTD) 295:330 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 4 47 theme secondary 476:484 arg1 structure 486:494 secondary structure 476:494 secondary structure 476:494 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 6 48 theme repeats 782:788 arg1 structure 766:774 the hairpin structure 754:774 the hairpin structure of T3 repeats 6 and 7 754:796 Disruption of the hairpin structure of T3 repeats 6 and 7 decreases protein secretion and stability. 15014436 2 49 theme C-terminal 299:308 arg1 CTD 327:329 CTD 327:329 CTD 327:329 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 2 49 theme C-terminal 299:308 arg1 domain 319:324 the C-terminal globular domain 295:324 the C-terminal globular domain (CTD) 295:330 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 9 50 theme human 1119:1123 arg1 disorders 1134:1142 two human skeletal disorders 1115:1142 two human skeletal disorders 1115:1142 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 6 51 theme T3 779:780 arg1 repeats 782:788 T3 repeats 6 and 7 779:796 T3 repeats 6 and 7 779:796 Disruption of the hairpin structure of T3 repeats 6 and 7 decreases protein secretion and stability. 15014436 7 52 theme calcium 925:931 arg1 loading 933:939 calcium loading 925:939 calcium loading 925:939 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 15014436 4 53 theme ions 539:542 arg1 core 523:526 a core 521:526 a core of calcium ions 521:542 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 5 54 theme conserved 707:715 arg1 sites 733:737 four strictly conserved calcium-binding sites 693:737 four strictly conserved calcium-binding sites 693:737 The CTD forms a lectin-like beta-sandwich and contains four strictly conserved calcium-binding sites. 15014436 2 55 theme TSPs 242:245 arg1 region 228:233 The most highly conserved region 202:233 The most highly conserved region of all TSPs 202:245 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 2 55 theme TSPs 242:245 arg1 repeats 283:289 the calcium-binding type 3 (T3) repeats 251:289 the calcium-binding type 3 (T3) repeats 251:289 The most highly conserved region of all TSPs are the calcium-binding type 3 (T3) repeats and the C-terminal globular domain (CTD). 15014436 4 56 theme calcium 531:537 arg1 ions 539:542 calcium ions 531:542 calcium ions 531:542 The T3 repeats lack secondary structure and are organised around a core of calcium ions; two DxDxDGxxDxxD motifs per repeat each encapsulate two calcium ions in a novel arrangement. 15014436 0 57 theme calcium 66:72 arg1 core 74:77 a novel calcium core 58:77 a novel calcium core 58:77 Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. 15014436 6 58 theme structure 766:774 arg1 Disruption 740:749 Disruption 740:749 Disruption of the hairpin structure of T3 repeats 6 and 7 740:796 Disruption of the hairpin structure of T3 repeats 6 and 7 decreases protein secretion and stability. 15014436 0 59 theme novel 60:64 arg1 core 74:77 a novel calcium core 58:77 a novel calcium core 58:77 Structure of a thrombospondin C-terminal fragment reveals a novel calcium core in the type 3 repeats. 15014436 9 60 theme tertiary 1174:1181 arg1 structure 1183:1191 the tertiary structure 1170:1191 the tertiary structure of the T3-CTD assembly 1170:1214 Mutations in the T3 repeats of TSP-5/COMP, which cause two human skeletal disorders, are predicted to disrupt the tertiary structure of the T3-CTD assembly. 15014436 7 61 from motif 888:892 arg1 repeat 900:905 T3 repeat 7 897:907 T3 repeat 7 897:907 The availability for cell attachment of an RGD motif in T3 repeat 7 is modulated by calcium loading. 19252480 9 0 theme LPS 1368:1370 arg1 LPS 1368:1370 LPS 1368:1370 LPS 1368:1370 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 9 0 theme LPS 1368:1370 arg1 groups 1358:1363 phosphate groups 1348:1363 phosphate groups of LPS 1348:1370 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 1 1 theme well-known 141:150 arg1 lipopolysaccharide 85:102 The lipopolysaccharide 81:102 The lipopolysaccharide (LPS) of Gram negative bacteria 81:134 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 1 1 theme well-known 141:150 arg1 inducer 152:158 a well-known inducer 139:158 a well-known inducer of the innate immune response 139:188 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 7 2 theme localized 984:992 arg1 change 1005:1010 localized structural change 984:1010 localized structural change 984:1010 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 10 3 theme TLR4-MD-2-LPS 1512:1524 arg1 structure 1526:1534 The TLR4-MD-2-LPS structure 1508:1534 The TLR4-MD-2-LPS structure 1508:1534 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 4 4 theme LPS 492:494 arg1 binding 496:502 LPS binding 492:502 LPS binding 492:502 LPS binding induced the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically. 19252480 7 5 theme hydrophilic 1067:1077 arg1 interactions 1079:1090 hydrophilic interactions 1067:1090 hydrophilic interactions with TLR4 1067:1100 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 4 6 theme multimer 550:557 arg1 formation 516:524 the formation 512:524 the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically 512:632 LPS binding induced the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically. 19252480 7 7 with interactions 1079:1090 arg1 TLR4 1097:1100 TLR4 1097:1100 TLR4 1097:1100 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 9 8 from TLR4 1493:1496 arg1 cluster 1451:1457 a cluster 1449:1457 a cluster of positively charged residues in TLR4 and MD-2 1449:1505 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 6 9 theme LPS 778:780 arg1 chains 768:773 the six lipid chains 754:773 the six lipid chains of LPS 754:780 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 8 10 with Comparison 1103:1112 arg1 structures 1123:1132 the structures 1119:1132 the structures of tetra-acylated antagonists bound to MD-2 1119:1176 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 10 11 theme mechanisms 1601:1610 arg1 versatility 1563:1573 the remarkable versatility 1548:1573 the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection 1548:1705 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 6 12 with interaction 897:907 arg1 phenylalanines 928:941 the conserved phenylalanines 914:941 the conserved phenylalanines of TLR4 914:949 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 3 13 theme crystal 473:479 arg1 structure 481:489 its crystal structure 469:489 its crystal structure 469:489 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 10 14 theme microbial 1687:1695 arg1 infection 1697:1705 diverse microbial infection 1679:1705 diverse microbial infection 1679:1705 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 8 15 theme tetra-acylated 1137:1150 arg1 antagonists 1152:1162 tetra-acylated antagonists 1137:1162 tetra-acylated antagonists bound to MD-2 1137:1176 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 4 16 theme complex 603:609 arg1 copies 575:580 two copies 571:580 two copies of the TLR4-MD-2-LPS complex arranged symmetrically 571:632 LPS binding induced the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically. 19252480 3 17 dep determined 458:467 arg1 understand 358:367 understand 358:367 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex 355:453 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 2 18 theme LPS 340:342 arg1 molecules 344:352 structurally diverse LPS molecules 319:352 structurally diverse LPS molecules 319:352 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 3 19 theme receptor 396:403 arg1 mechanism 416:424 the ligand specificity and receptor activation mechanism 369:424 mechanism 416:424 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 10 20 theme remarkable 1552:1561 arg1 versatility 1563:1573 the remarkable versatility 1548:1573 the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection 1548:1705 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 6 21 theme lipid 762:766 arg1 chains 768:773 the six lipid chains 754:773 the six lipid chains of LPS 754:780 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 9 22 theme residues 1481:1488 arg1 cluster 1451:1457 a cluster 1449:1457 a cluster of positively charged residues in TLR4 and MD-2 1449:1505 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 3 23 theme activation 405:414 arg1 mechanism 416:424 the ligand specificity and receptor activation mechanism 369:424 mechanism 416:424 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 0 24 theme structural 4:13 arg1 basis 15:19 The structural basis 0:19 The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex 0:78 The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. 19252480 4 25 theme m-shaped 532:539 arg1 multimer 550:557 an m-shaped receptor multimer 529:557 an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically 529:632 LPS binding induced the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically. 19252480 7 26 theme core 1030:1033 arg1 interface 1047:1055 this core hydrophobic interface 1025:1055 this core hydrophobic interface 1025:1055 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 2 27 from pattern 307:313 arg1 molecules 344:352 structurally diverse LPS molecules 319:352 structurally diverse LPS molecules 319:352 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 7 28 theme structural 994:1003 arg1 change 1005:1010 localized structural change 984:1010 localized structural change 984:1010 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 1 29 theme innate 167:172 arg1 response 181:188 the innate immune response 163:188 the innate immune response 163:188 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 8 30 theme antagonists 1152:1162 arg1 structures 1123:1132 the structures 1119:1132 the structures of tetra-acylated antagonists bound to MD-2 1119:1176 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 6 31 theme conserved 918:926 arg1 phenylalanines 928:941 the conserved phenylalanines 914:941 the conserved phenylalanines of TLR4 914:949 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 7 32 theme hydrophobic 1035:1045 arg1 interface 1047:1055 this core hydrophobic interface 1025:1055 this core hydrophobic interface 1025:1055 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 2 33 theme diverse 332:338 arg1 molecules 344:352 structurally diverse LPS molecules 319:352 structurally diverse LPS molecules 319:352 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 10 34 theme recognition 1589:1599 arg1 mechanisms 1601:1610 the ligand recognition mechanisms 1578:1610 the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection 1578:1705 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 6 35 theme chains 768:773 arg1 deep 793:796 buried deep 786:796 buried deep inside the pocket 786:814 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 6 35 theme chains 768:773 arg1 chains 768:773 the six lipid chains 754:773 the six lipid chains of LPS 754:780 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 6 35 theme chains 768:773 arg1 Five 746:749 Five 746:749 Five 746:749 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 1 36 theme response 181:188 arg1 lipopolysaccharide 85:102 The lipopolysaccharide 81:102 The lipopolysaccharide (LPS) of Gram negative bacteria 81:134 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 1 36 theme response 181:188 arg1 inducer 152:158 a well-known inducer 139:158 a well-known inducer of the innate immune response 139:188 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 10 37 theme ligand 1582:1587 arg1 mechanisms 1601:1610 the ligand recognition mechanisms 1578:1610 the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection 1578:1705 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 4 38 theme receptor 541:548 arg1 multimer 550:557 an m-shaped receptor multimer 529:557 an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically 529:632 LPS binding induced the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically. 19252480 6 39 theme remaining 824:832 arg1 chain 834:838 the remaining chain 820:838 the remaining chain 820:838 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 3 40 theme TLR4-MD-2-LPS 433:445 arg1 complex 447:453 the TLR4-MD-2-LPS complex 429:453 the TLR4-MD-2-LPS complex 429:453 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 0 41 theme recognition 43:53 arg1 basis 15:19 The structural basis 0:19 The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex 0:78 The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. 19252480 2 42 theme common 299:304 arg1 pattern 307:313 a common 'pattern' 297:314 a common 'pattern' in structurally diverse LPS molecules 297:352 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 9 43 from MD-2 1502:1505 arg1 cluster 1451:1457 a cluster 1449:1457 a cluster of positively charged residues in TLR4 and MD-2 1449:1505 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 1 44 theme immune 174:179 arg1 response 181:188 the innate immune response 163:188 the innate immune response 163:188 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 8 45 theme glucosamine 1251:1261 arg1 backbone 1263:1270 the phosphorylated glucosamine backbone 1232:1270 the phosphorylated glucosamine backbone by approximately 5 A 1232:1291 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 0 46 theme lipopolysaccharide 24:41 arg1 recognition 43:53 lipopolysaccharide recognition 24:53 lipopolysaccharide recognition 24:53 The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. 19252480 6 47 theme hydrophobic 885:895 arg1 interaction 897:907 a hydrophobic interaction 883:907 a hydrophobic interaction with the conserved phenylalanines of TLR4 883:949 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 3 48 theme ligand 373:378 arg1 specificity 380:390 the ligand specificity and receptor activation mechanism 369:424 specificity 380:390 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 6 49 theme TLR4 946:949 arg1 phenylalanines 928:941 the conserved phenylalanines 914:941 the conserved phenylalanines of TLR4 914:949 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 8 50 theme phosphorylated 1236:1249 arg1 backbone 1263:1270 the phosphorylated glucosamine backbone 1232:1270 the phosphorylated glucosamine backbone by approximately 5 A 1232:1291 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 8 51 theme other 1197:1201 arg1 chains 1209:1214 two other lipid chains 1193:1214 two other lipid chains in LPS 1193:1221 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 9 52 theme ionic 1425:1429 arg1 interactions 1431:1442 ionic interactions 1425:1442 ionic interactions 1425:1442 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 5 53 theme large 656:660 arg1 pocket 674:679 a large hydrophobic pocket 654:679 a large hydrophobic pocket in MD-2 654:687 LPS interacts with a large hydrophobic pocket in MD-2 and directly bridges the two components of the multimer. 19252480 5 54 theme multimer 736:743 arg1 components 718:727 the two components 710:727 the two components of the multimer 710:743 LPS interacts with a large hydrophobic pocket in MD-2 and directly bridges the two components of the multimer. 19252480 9 55 theme charged 1473:1479 arg1 residues 1481:1488 positively charged residues 1462:1488 positively charged residues in TLR4 and MD-2 1462:1505 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 9 56 theme phosphate 1348:1356 arg1 LPS 1368:1370 LPS 1368:1370 LPS 1368:1370 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 9 56 theme phosphate 1348:1356 arg1 groups 1358:1363 phosphate groups 1348:1363 phosphate groups of LPS 1348:1370 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 5 57 theme hydrophobic 662:672 arg1 pocket 674:679 a large hydrophobic pocket 654:679 a large hydrophobic pocket in MD-2 654:687 LPS interacts with a large hydrophobic pocket in MD-2 and directly bridges the two components of the multimer. 19252480 2 58 theme receptor 201:208 arg1 TLR 211:213 Toll-like receptor (TLR) 4 191:216 Toll-like receptor (TLR) 4 191:216 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 8 59 from chains 1209:1214 arg1 LPS 1219:1221 LPS 1219:1221 LPS 1219:1221 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 1 60 theme Gram 113:116 arg1 bacteria 127:134 Gram negative bacteria 113:134 Gram negative bacteria 113:134 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 9 61 from residues 1481:1488 arg1 MD-2 1502:1505 MD-2 1502:1505 MD-2 1502:1505 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 9 61 from residues 1481:1488 arg1 TLR4 1493:1496 TLR4 1493:1496 TLR4 1493:1496 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 7 62 theme MD-2 969:972 arg1 loop 961:964 The F126 loop 952:964 The F126 loop of MD-2 952:972 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 2 63 theme Toll-like 191:199 arg1 TLR 211:213 Toll-like receptor (TLR) 4 191:216 Toll-like receptor (TLR) 4 191:216 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 9 64 theme structural 1324:1333 arg1 shift 1335:1339 This structural shift 1319:1339 This structural shift 1319:1339 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 7 65 theme F126 956:959 arg1 loop 961:964 The F126 loop 952:964 The F126 loop of MD-2 952:972 The F126 loop of MD-2 undergoes localized structural change and supports this core hydrophobic interface by making hydrophilic interactions with TLR4. 19252480 1 66 theme negative 118:125 arg1 bacteria 127:134 Gram negative bacteria 113:134 Gram negative bacteria 113:134 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 5 67 from pocket 674:679 arg1 MD-2 684:687 MD-2 684:687 MD-2 684:687 LPS interacts with a large hydrophobic pocket in MD-2 and directly bridges the two components of the multimer. 19252480 9 68 from cluster 1451:1457 arg1 MD-2 1502:1505 MD-2 1502:1505 MD-2 1502:1505 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 9 68 from cluster 1451:1457 arg1 TLR4 1493:1496 TLR4 1493:1496 TLR4 1493:1496 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 9 69 theme receptor 1389:1396 arg1 multimerization 1398:1412 receptor multimerization 1389:1412 receptor multimerization 1389:1412 This structural shift allows phosphate groups of LPS to contribute to receptor multimerization by forming ionic interactions with a cluster of positively charged residues in TLR4 and MD-2. 19252480 2 70 theme differentiation 230:244 arg1 MD-2 256:259 MD-2 256:259 MD-2 256:259 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 2 70 theme differentiation 230:244 arg1 factor 246:251 myeloid differentiation factor 2 222:253 myeloid differentiation factor 2 (MD-2) 222:260 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 1 71 theme bacteria 127:134 arg1 LPS 105:107 LPS 105:107 LPS 105:107 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 1 71 theme bacteria 127:134 arg1 lipopolysaccharide 85:102 The lipopolysaccharide 81:102 The lipopolysaccharide (LPS) of Gram negative bacteria 81:134 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 1 71 theme bacteria 127:134 arg1 inducer 152:158 a well-known inducer 139:158 a well-known inducer of the innate immune response 139:188 The lipopolysaccharide (LPS) of Gram negative bacteria is a well-known inducer of the innate immune response. 19252480 8 72 theme solvent 1305:1311 arg1 area 1313:1316 the solvent area 1301:1316 the solvent area 1301:1316 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 3 73 theme complex 447:453 arg1 specificity 380:390 the ligand specificity and receptor activation mechanism 369:424 specificity 380:390 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 3 73 theme complex 447:453 arg1 mechanism 416:424 the ligand specificity and receptor activation mechanism 369:424 mechanism 416:424 To understand the ligand specificity and receptor activation mechanism of the TLR4-MD-2-LPS complex we determined its crystal structure. 19252480 6 74 theme buried 786:791 arg1 deep 793:796 buried deep 786:796 buried deep inside the pocket 786:814 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 6 74 theme buried 786:791 arg1 chains 768:773 the six lipid chains 754:773 the six lipid chains of LPS 754:780 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 6 74 theme buried 786:791 arg1 Five 746:749 Five 746:749 Five 746:749 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 2 75 theme myeloid 222:228 arg1 MD-2 256:259 MD-2 256:259 MD-2 256:259 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 2 75 theme myeloid 222:228 arg1 factor 246:251 myeloid differentiation factor 2 222:253 myeloid differentiation factor 2 (MD-2) 222:260 Toll-like receptor (TLR) 4 and myeloid differentiation factor 2 (MD-2) form a heterodimer that recognizes a common 'pattern' in structurally diverse LPS molecules. 19252480 10 76 theme diverse 1679:1685 arg1 infection 1697:1705 diverse microbial infection 1679:1705 diverse microbial infection 1679:1705 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 8 77 theme lipid 1203:1207 arg1 chains 1209:1214 two other lipid chains 1193:1214 two other lipid chains in LPS 1193:1221 Comparison with the structures of tetra-acylated antagonists bound to MD-2 indicates that two other lipid chains in LPS displace the phosphorylated glucosamine backbone by approximately 5 A towards the solvent area. 19252480 4 78 theme TLR4-MD-2-LPS 589:601 arg1 complex 603:609 the TLR4-MD-2-LPS complex 585:609 the TLR4-MD-2-LPS complex arranged symmetrically 585:632 LPS binding induced the formation of an m-shaped receptor multimer composed of two copies of the TLR4-MD-2-LPS complex arranged symmetrically. 19252480 0 79 theme TLR4-MD-2 62:70 arg1 complex 72:78 the TLR4-MD-2 complex 58:78 the TLR4-MD-2 complex 58:78 The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. 19252480 6 80 theme MD-2 869:872 arg1 surface 858:864 the surface 854:864 the surface of MD-2 854:872 Five of the six lipid chains of LPS are buried deep inside the pocket and the remaining chain is exposed to the surface of MD-2, forming a hydrophobic interaction with the conserved phenylalanines of TLR4. 19252480 10 81 theme TLR 1628:1630 arg1 essential 1649:1657 essential 1649:1657 essential 1649:1657 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 19252480 10 81 theme TLR 1628:1630 arg1 family 1632:1637 the TLR family 1624:1637 the TLR family 1624:1637 The TLR4-MD-2-LPS structure illustrates the remarkable versatility of the ligand recognition mechanisms employed by the TLR family, which is essential for defence against diverse microbial infection. 17803912 3 0 dep TLR4 355:358 arg1 the 345:347 the 345:347 the 345:347 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 6 1 theme central 650:656 arg1 domain 658:663 the central domain 646:663 the central domain 646:663 The beta sheet of the central domain shows unusually small radii and large twist angles. 17803912 9 2 from experiments 919:929 arg1 TLR4 943:946 TLR4 943:946 TLR4 943:946 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 9 2 from experiments 919:929 arg1 MD-2 934:937 MD-2 934:937 MD-2 934:937 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 5 3 theme central 595:601 arg1 domains 619:625 N-terminal, central, and C-terminal domains 583:625 N-terminal, central, and C-terminal domains 583:625 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 8 4 with interaction 794:804 arg1 Eritoran 811:818 Eritoran 811:818 Eritoran 811:818 The interaction with Eritoran is mediated by a hydrophobic internal pocket in MD-2. 17803912 5 5 theme LRR 553:555 arg1 family 557:562 the LRR family 549:562 the LRR family 549:562 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 9 6 theme TLR4-MD-2 971:979 arg1 dimerization 981:992 TLR4-MD-2 dimerization 971:992 TLR4-MD-2 dimerization induced by LPS 971:1007 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 3 7 theme mouse 349:353 arg1 TLR4 355:358 mouse TLR4 349:358 mouse TLR4 349:358 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 4 8 theme VLR 441:443 arg1 hybrids 407:413 hybrids 407:413 hybrids of human TLR4 and hagfish VLR 407:443 We also produced a series of hybrids of human TLR4 and hagfish VLR and determined their structures with and without bound MD-2 and Eritoran. 17803912 5 9 theme family 557:562 arg1 member 539:544 an atypical member 527:544 an atypical member of the LRR family 527:562 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 5 9 theme family 557:562 arg1 TLR4 519:522 TLR4 519:522 TLR4 519:522 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 2 10 theme TLR4-MD-2 265:273 arg1 complex 275:281 the TLR4-MD-2 complex 261:281 the TLR4-MD-2 complex 261:281 Eritoran is an analog of LPS that antagonizes its activity by binding to the TLR4-MD-2 complex. 17803912 7 11 theme domains 781:787 arg1 surface 743:749 the concave surface 731:749 the concave surface of the N-terminal and central domains 731:787 MD-2 binds to the concave surface of the N-terminal and central domains. 17803912 7 12 theme central 773:779 arg1 domains 781:787 the N-terminal and central domains 754:787 the N-terminal and central domains 754:787 MD-2 binds to the concave surface of the N-terminal and central domains. 17803912 3 13 theme ectodomain 331:340 arg1 structure 302:310 the structure 298:310 the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex 298:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 4 14 theme hagfish 433:439 arg1 VLR 441:443 hagfish VLR 433:443 hagfish VLR 433:443 We also produced a series of hybrids of human TLR4 and hagfish VLR and determined their structures with and without bound MD-2 and Eritoran. 17803912 0 15 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 0:83 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 1 16 theme Gram-negative 164:176 arg1 bacteria 178:185 Gram-negative bacteria 164:185 Gram-negative bacteria 164:185 TLR4 and MD-2 form a heterodimer that recognizes LPS (lipopolysaccharide) from Gram-negative bacteria. 17803912 3 17 theme MD-2 364:367 arg1 complex 369:375 MD-2 complex 364:375 MD-2 complex 364:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 9 18 from analysis 894:901 arg1 TLR4 943:946 TLR4 943:946 TLR4 943:946 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 9 18 from analysis 894:901 arg1 MD-2 934:937 MD-2 934:937 MD-2 934:937 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 5 19 theme atypical 530:537 arg1 member 539:544 an atypical member 527:544 an atypical member of the LRR family 527:562 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 5 19 theme atypical 530:537 arg1 TLR4 519:522 TLR4 519:522 TLR4 519:522 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 6 20 theme beta 632:635 arg1 sheet 637:641 The beta sheet 628:641 The beta sheet of the central domain 628:663 The beta sheet of the central domain shows unusually small radii and large twist angles. 17803912 3 21 theme complex 369:375 arg1 TLR4 355:358 mouse TLR4 349:358 mouse TLR4 349:358 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 3 21 theme complex 369:375 arg1 complex 369:375 MD-2 complex 364:375 MD-2 complex 364:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 3 21 theme complex 369:375 arg1 ectodomain 331:340 the full-length ectodomain 315:340 the full-length ectodomain of the mouse TLR4 and MD-2 complex 315:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 5 22 theme N-terminal 583:592 arg1 domains 619:625 N-terminal, central, and C-terminal domains 583:625 N-terminal, central, and C-terminal domains 583:625 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 3 23 theme TLR4 355:358 arg1 TLR4 355:358 mouse TLR4 349:358 mouse TLR4 349:358 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 3 23 theme TLR4 355:358 arg1 complex 369:375 MD-2 complex 364:375 MD-2 complex 364:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 3 23 theme TLR4 355:358 arg1 ectodomain 331:340 the full-length ectodomain 315:340 the full-length ectodomain of the mouse TLR4 and MD-2 complex 315:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 7 24 theme concave 735:741 arg1 surface 743:749 the concave surface 731:749 the concave surface of the N-terminal and central domains 731:787 MD-2 binds to the concave surface of the N-terminal and central domains. 17803912 6 25 theme twist 703:707 arg1 angles 709:714 large twist angles 697:714 large twist angles 697:714 The beta sheet of the central domain shows unusually small radii and large twist angles. 17803912 7 26 theme N-terminal 758:767 arg1 domains 781:787 the N-terminal and central domains 754:787 the N-terminal and central domains 754:787 MD-2 binds to the concave surface of the N-terminal and central domains. 17803912 3 27 theme full-length 319:329 arg1 TLR4 355:358 mouse TLR4 349:358 mouse TLR4 349:358 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 3 27 theme full-length 319:329 arg1 complex 369:375 MD-2 complex 364:375 MD-2 complex 364:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 3 27 theme full-length 319:329 arg1 ectodomain 331:340 the full-length ectodomain 315:340 the full-length ectodomain of the mouse TLR4 and MD-2 complex 315:375 We determined the structure of the full-length ectodomain of the mouse TLR4 and MD-2 complex. 17803912 4 28 theme human 418:422 arg1 TLR4 424:427 human TLR4 418:427 human TLR4 418:427 We also produced a series of hybrids of human TLR4 and hagfish VLR and determined their structures with and without bound MD-2 and Eritoran. 17803912 0 29 theme complex 35:41 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 0:83 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 0 30 theme TLR4-MD-2 25:33 arg1 complex 35:41 the TLR4-MD-2 complex 21:41 the TLR4-MD-2 complex 21:41 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 4 31 theme bound 494:498 arg1 MD-2 500:503 MD-2 500:503 MD-2 500:503 We also produced a series of hybrids of human TLR4 and hagfish VLR and determined their structures with and without bound MD-2 and Eritoran. 17803912 6 32 theme large 697:701 arg1 angles 709:714 large twist angles 697:714 large twist angles 697:714 The beta sheet of the central domain shows unusually small radii and large twist angles. 17803912 0 33 theme bound 48:52 arg1 Eritoran 75:82 bound endotoxin antagonist Eritoran 48:82 bound endotoxin antagonist Eritoran 48:82 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 5 34 theme C-terminal 608:617 arg1 domains 619:625 N-terminal, central, and C-terminal domains 583:625 N-terminal, central, and C-terminal domains 583:625 TLR4 is an atypical member of the LRR family and is composed of N-terminal, central, and C-terminal domains. 17803912 4 35 theme TLR4 424:427 arg1 hybrids 407:413 hybrids 407:413 hybrids of human TLR4 and hagfish VLR 407:443 We also produced a series of hybrids of human TLR4 and hagfish VLR and determined their structures with and without bound MD-2 and Eritoran. 17803912 2 36 theme LPS 213:215 arg1 Eritoran 188:195 Eritoran 188:195 Eritoran 188:195 Eritoran is an analog of LPS that antagonizes its activity by binding to the TLR4-MD-2 complex. 17803912 2 36 theme LPS 213:215 arg1 analog 203:208 an analog 200:208 an analog of LPS that antagonizes its activity by binding to the TLR4-MD-2 complex 200:281 Eritoran is an analog of LPS that antagonizes its activity by binding to the TLR4-MD-2 complex. 17803912 8 37 theme hydrophobic 837:847 arg1 pocket 858:863 a hydrophobic internal pocket 835:863 a hydrophobic internal pocket in MD-2 835:871 The interaction with Eritoran is mediated by a hydrophobic internal pocket in MD-2. 17803912 0 38 with structure 8:16 arg1 Eritoran 75:82 bound endotoxin antagonist Eritoran 48:82 bound endotoxin antagonist Eritoran 48:82 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 6 39 theme small 681:685 arg1 radii 687:691 unusually small radii 671:691 unusually small radii 671:691 The beta sheet of the central domain shows unusually small radii and large twist angles. 17803912 4 40 theme hybrids 407:413 arg1 series 397:402 a series 395:402 a series of hybrids of human TLR4 and hagfish VLR 395:443 We also produced a series of hybrids of human TLR4 and hagfish VLR and determined their structures with and without bound MD-2 and Eritoran. 17803912 8 41 theme internal 849:856 arg1 pocket 858:863 a hydrophobic internal pocket 835:863 a hydrophobic internal pocket in MD-2 835:871 The interaction with Eritoran is mediated by a hydrophobic internal pocket in MD-2. 17803912 1 42 from bacteria 178:185 arg1 lipopolysaccharide 139:156 lipopolysaccharide 139:156 lipopolysaccharide 139:156 TLR4 and MD-2 form a heterodimer that recognizes LPS (lipopolysaccharide) from Gram-negative bacteria. 17803912 1 42 from bacteria 178:185 arg1 LPS 134:136 LPS 134:136 LPS (lipopolysaccharide) from Gram-negative bacteria 134:185 TLR4 and MD-2 form a heterodimer that recognizes LPS (lipopolysaccharide) from Gram-negative bacteria. 17803912 0 43 theme antagonist 64:73 arg1 Eritoran 75:82 bound endotoxin antagonist Eritoran 48:82 bound endotoxin antagonist Eritoran 48:82 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 8 44 from pocket 858:863 arg1 MD-2 868:871 MD-2 868:871 MD-2 868:871 The interaction with Eritoran is mediated by a hydrophobic internal pocket in MD-2. 17803912 0 45 theme endotoxin 54:62 arg1 Eritoran 75:82 bound endotoxin antagonist Eritoran 48:82 bound endotoxin antagonist Eritoran 48:82 Crystal structure of the TLR4-MD-2 complex with bound endotoxin antagonist Eritoran. 17803912 9 46 theme mutagenesis 907:917 arg1 experiments 919:929 mutagenesis experiments 907:929 mutagenesis experiments on MD-2 and TLR4 907:946 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 6 47 theme domain 658:663 arg1 sheet 637:641 The beta sheet 628:641 The beta sheet of the central domain 628:663 The beta sheet of the central domain shows unusually small radii and large twist angles. 17803912 9 48 theme structural 883:892 arg1 analysis 894:901 structural analysis 883:901 structural analysis 883:901 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 17803912 9 49 theme dimerization 981:992 arg1 model 962:966 a model 960:966 a model of TLR4-MD-2 dimerization induced by LPS 960:1007 Based on structural analysis and mutagenesis experiments on MD-2 and TLR4, we propose a model of TLR4-MD-2 dimerization induced by LPS. 22363519 0 0 theme decoy 65:69 arg1 receptor 71:78 a Toll-like receptor 4 (TLR4) decoy receptor 35:78 a Toll-like receptor 4 (TLR4) decoy receptor 35:78 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 7 1 theme single 1266:1271 arg1 mutations 1282:1290 single positive mutations 1266:1290 single positive mutations 1266:1290 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 1 2 theme Repeat 129:134 arg1 proteins 136:143 Repeat proteins 129:143 Repeat proteins 129:143 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 8 3 theme binding 1627:1633 arg1 affinities 1635:1644 the binding affinities 1623:1644 the binding affinities 1623:1644 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 8 4 theme Molecular 1435:1443 arg1 simulations 1454:1464 Molecular dynamics simulations 1435:1464 Molecular dynamics simulations 1435:1464 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 5 5 theme wild-type 1059:1067 arg1 receptor 1075:1082 the wild-type decoy receptor 1055:1082 the wild-type decoy receptor 1055:1082 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 5 6 theme receptor 866:873 arg1 structure 843:851 the complex crystal structure 823:851 the complex crystal structure of the decoy receptor with MD2 823:882 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 2 7 theme affinity 368:375 arg1 maturation 377:386 affinity maturation 368:386 affinity maturation of repeat proteins 368:405 Nonetheless, engineering interaction interface and understanding molecular basis for affinity maturation of repeat proteins still remain a challenge. 22363519 5 8 from substitutions 921:933 arg1 receptor 948:955 the decoy receptor 938:955 the decoy receptor 938:955 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 8 9 theme major 1575:1579 arg1 effect 1515:1520 an additive effect 1503:1520 an additive effect by two mutations occurring at nearby modules 1503:1565 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 8 9 theme major 1575:1579 arg1 contributor 1581:1591 the major contributor 1571:1591 the major contributor to the remarkable increase in the binding affinities 1571:1644 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 5 10 theme decoy 1069:1073 arg1 receptor 1075:1082 the wild-type decoy receptor 1055:1082 the wild-type decoy receptor 1055:1082 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 7 11 theme 565-fold 1365:1372 arg1 affinities 1389:1398 higher binding affinities 1374:1398 3000- and 565-fold higher binding affinities than the wild-type decoy receptor 1355:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 1 12 theme unique 255:260 arg1 features 273:280 their unique structural features 249:280 their unique structural features 249:280 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 0 13 theme receptor 71:78 arg1 design 25:30 Structure-based rational design 0:30 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 0:127 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 7 14 contain have 1344:1347 arg1 mutants 1322:1328 two double mutants 1311:1328 two double mutants 1311:1328 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 7 14 contain have 1344:1347 arg2 affinities 1389:1398 higher binding affinities 1374:1398 3000- and 565-fold higher binding affinities than the wild-type decoy receptor 1355:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 6 15 theme variants 1214:1221 arg1 structures 1189:1198 the crystal structures 1177:1198 the crystal structures of the single variants 1177:1221 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 5 16 dep affinity 1004:1011 arg1 D 1016:1016 D 1016:1016 D 1016:1016 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 5 16 dep affinity 1004:1011 arg1 K 1014:1014 K 1014:1014 K(D) 1014:1017 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 3 17 theme binding 514:520 arg1 affinity 522:529 high binding affinity 509:529 high binding affinity for a target protein 509:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 1 18 theme structural 262:271 arg1 features 273:280 their unique structural features 249:280 their unique structural features 249:280 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 0 19 theme binding 90:96 arg1 affinity 98:105 high binding affinity 85:105 high binding affinity for a target protein 85:126 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 8 20 theme energetic 1470:1478 arg1 analysis 1480:1487 energetic analysis 1470:1487 energetic analysis 1470:1487 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 6 21 theme single 1207:1212 arg1 variants 1214:1221 the single variants 1203:1221 the single variants 1203:1221 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 4 22 theme Toll-like 582:590 arg1 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 22 theme Toll-like 582:590 arg1 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 3 23 theme target 537:542 arg1 protein 544:550 a target protein 535:550 a target protein 535:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 5 24 theme complex 827:833 arg1 structure 843:851 the complex crystal structure 823:851 the complex crystal structure of the decoy receptor with MD2 823:882 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 0 25 theme high 85:88 arg1 affinity 98:105 high binding affinity 85:105 high binding affinity for a target protein 85:126 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 5 26 theme magnitude 1033:1041 arg1 one-order 1020:1028 one-order 1020:1028 one-order of magnitude 1020:1041 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 3 27 with protein 496:502 arg1 affinity 522:529 high binding affinity 509:529 high binding affinity for a target protein 509:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 4 28 used used 674:677 arg2 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 28 used used 674:677 arg2 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 29 theme binding 752:758 arg1 affinity 760:767 the binding affinity 748:767 the binding affinity for myeloid differentiation protein 2 (MD2) 748:811 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 3 30 theme structure-based 452:466 arg1 design 477:482 a structure-based rational design 450:482 a structure-based rational design of a repeat protein with high binding affinity for a target protein 450:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 0 31 theme rational 16:23 arg1 design 25:30 Structure-based rational design 0:30 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 0:127 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 5 32 theme higher 1043:1048 arg1 affinity 1004:1011 a binding affinity 994:1011 a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor 994:1082 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 8 33 theme additive 1506:1513 arg1 effect 1515:1520 an additive effect 1503:1520 an additive effect by two mutations occurring at nearby modules 1503:1565 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 8 33 theme additive 1506:1513 arg1 contributor 1581:1591 the major contributor 1571:1591 the major contributor to the remarkable increase in the binding affinities 1571:1644 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 6 34 theme interacting 1089:1099 arg1 modes 1101:1105 modes 1101:1105 modes 1101:1105 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 4 35 theme leucine-rich 636:647 arg1 LRR 657:659 LRR 657:659 LRR 657:659 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 35 theme leucine-rich 636:647 arg1 repeat 649:654 leucine-rich repeat 636:654 leucine-rich repeat (LRR) modules 636:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 0 36 theme Structure-based 0:14 arg1 design 25:30 Structure-based rational design 0:30 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 0:127 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 4 37 theme model 558:562 arg1 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 37 theme model 558:562 arg1 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 38 theme interaction 688:698 arg1 interface 700:708 its interaction interface 684:708 its interaction interface 684:708 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 5 39 theme crystal 835:841 arg1 structure 843:851 the complex crystal structure 823:851 the complex crystal structure of the decoy receptor with MD2 823:882 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 2 40 theme proteins 398:405 arg1 maturation 377:386 affinity maturation 368:386 affinity maturation of repeat proteins 368:405 Nonetheless, engineering interaction interface and understanding molecular basis for affinity maturation of repeat proteins still remain a challenge. 22363519 7 41 theme binding 1381:1387 arg1 affinities 1389:1398 higher binding affinities 1374:1398 3000- and 565-fold higher binding affinities than the wild-type decoy receptor 1355:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 2 42 theme interaction 308:318 arg1 interface 320:328 interaction interface 308:328 interaction interface 308:328 Nonetheless, engineering interaction interface and understanding molecular basis for affinity maturation of repeat proteins still remain a challenge. 22363519 7 43 theme binding 1248:1254 arg1 affinity 1256:1263 the binding affinity 1244:1263 the binding affinity 1244:1263 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 5 44 theme single 903:908 arg1 substitutions 921:933 single amino acid substitutions 903:933 single amino acid substitutions in the decoy receptor 903:955 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 7 45 theme 3000- 1355:1359 arg1 affinities 1389:1398 higher binding affinities 1374:1398 3000- and 565-fold higher binding affinities than the wild-type decoy receptor 1355:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 5 46 theme acid 916:919 arg1 substitutions 921:933 single amino acid substitutions 903:933 single amino acid substitutions in the decoy receptor 903:955 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 0 47 theme Toll-like 37:45 arg1 receptor 47:54 Toll-like receptor 4 37:56 a Toll-like receptor 4 (TLR4) decoy receptor 35:78 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 0 47 theme Toll-like 37:45 arg1 TLR4 59:62 TLR4 59:62 TLR4 59:62 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 4 48 theme repeat 564:569 arg1 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 48 theme repeat 564:569 arg1 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 0 49 theme target 113:118 arg1 protein 120:126 a target protein 111:126 a target protein 111:126 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 3 50 theme rational 468:475 arg1 design 477:482 a structure-based rational design 450:482 a structure-based rational design of a repeat protein with high binding affinity for a target protein 450:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 6 51 theme crystal 1181:1187 arg1 structures 1189:1198 the crystal structures 1177:1198 the crystal structures of the single variants 1177:1221 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 1 52 theme much 173:176 arg1 attention 178:186 much attention 173:186 much attention 173:186 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 1 52 theme much 173:176 arg1 scaffolds 203:211 alternative scaffolds 191:211 alternative scaffolds to immunoglobulin antibodies due to their unique structural features 191:280 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 1 53 theme alternative 191:201 arg1 attention 178:186 much attention 173:186 much attention 173:186 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 1 53 theme alternative 191:201 arg1 scaffolds 203:211 alternative scaffolds 191:211 alternative scaffolds to immunoglobulin antibodies due to their unique structural features 191:280 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 7 54 theme positive 1273:1280 arg1 mutations 1282:1290 single positive mutations 1266:1290 single positive mutations 1266:1290 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 7 55 theme wild-type 1409:1417 arg1 receptor 1425:1432 the wild-type decoy receptor 1405:1432 the wild-type decoy receptor 1405:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 5 56 with structure 843:851 arg1 MD2 880:882 MD2 880:882 MD2 880:882 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 0 57 with design 25:30 arg1 affinity 98:105 high binding affinity 85:105 high binding affinity for a target protein 85:126 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 8 58 theme nearby 1552:1557 arg1 modules 1559:1565 nearby modules 1552:1565 nearby modules 1552:1565 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 0 59 theme receptor 47:54 arg1 receptor 71:78 a Toll-like receptor 4 (TLR4) decoy receptor 35:78 a Toll-like receptor 4 (TLR4) decoy receptor 35:78 Structure-based rational design of a Toll-like receptor 4 (TLR4) decoy receptor with high binding affinity for a target protein. 22363519 3 60 theme repeat 489:494 arg1 protein 496:502 a repeat protein 487:502 a repeat protein with high binding affinity for a target protein 487:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 5 61 theme binding 996:1002 arg1 affinity 1004:1011 a binding affinity 994:1011 a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor 994:1082 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 7 62 theme double 1315:1320 arg1 mutants 1322:1328 two double mutants 1311:1328 two double mutants 1311:1328 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 3 63 theme high 509:512 arg1 affinity 522:529 high binding affinity 509:529 high binding affinity for a target protein 509:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 7 64 theme decoy 1419:1423 arg1 receptor 1425:1432 the wild-type decoy receptor 1405:1432 the wild-type decoy receptor 1405:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 6 65 theme individual 1128:1137 arg1 residues 1139:1146 individual residues 1128:1146 individual residues 1128:1146 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 8 66 theme remarkable 1600:1609 arg1 increase 1611:1618 the remarkable increase 1596:1618 the remarkable increase in the binding affinities 1596:1644 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 8 67 from increase 1611:1618 arg1 affinities 1635:1644 the binding affinities 1623:1644 the binding affinities 1623:1644 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 4 68 theme myeloid 773:779 arg1 protein 797:803 myeloid differentiation protein 2 773:805 myeloid differentiation protein 2 (MD2) 773:811 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 68 theme myeloid 773:779 arg1 MD2 808:810 MD2 808:810 MD2 808:810 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 2 69 theme molecular 348:356 arg1 basis 358:362 molecular basis 348:362 molecular basis for affinity maturation of repeat proteins 348:405 Nonetheless, engineering interaction interface and understanding molecular basis for affinity maturation of repeat proteins still remain a challenge. 22363519 5 70 theme amino 910:914 arg1 substitutions 921:933 single amino acid substitutions 903:933 single amino acid substitutions in the decoy receptor 903:955 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 4 71 theme TLR4 603:606 arg1 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 71 theme TLR4 603:606 arg1 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 5 72 theme decoy 942:946 arg1 receptor 948:955 the decoy receptor 938:955 the decoy receptor 938:955 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 22363519 3 73 theme protein 496:502 arg1 design 477:482 a structure-based rational design 450:482 a structure-based rational design of a repeat protein with high binding affinity for a target protein 450:550 Here, we present a structure-based rational design of a repeat protein with high binding affinity for a target protein. 22363519 1 74 theme immunoglobulin 216:229 arg1 antibodies 231:240 immunoglobulin antibodies 216:240 immunoglobulin antibodies due to their unique structural features 216:280 Repeat proteins are increasingly attracting much attention as alternative scaffolds to immunoglobulin antibodies due to their unique structural features. 22363519 7 75 theme higher 1374:1379 arg1 affinities 1389:1398 higher binding affinities 1374:1398 3000- and 565-fold higher binding affinities than the wild-type decoy receptor 1355:1432 To further increase the binding affinity, single positive mutations were combined, and two double mutants were shown to have about 3000- and 565-fold higher binding affinities than the wild-type decoy receptor. 22363519 6 76 theme residues 1139:1146 arg1 contributions 1111:1123 contributions 1111:1123 contributions 1111:1123 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 6 76 theme residues 1139:1146 arg1 modes 1101:1105 modes 1101:1105 modes 1101:1105 The interacting modes and contributions of individual residues were elucidated by analyzing the crystal structures of the single variants. 22363519 2 77 theme repeat 391:396 arg1 proteins 398:405 repeat proteins 391:405 repeat proteins 391:405 Nonetheless, engineering interaction interface and understanding molecular basis for affinity maturation of repeat proteins still remain a challenge. 22363519 4 78 theme repeat 649:654 arg1 modules 662:668 leucine-rich repeat (LRR) modules 636:668 leucine-rich repeat (LRR) modules 636:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 79 theme decoy 609:613 arg1 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 79 theme decoy 609:613 arg1 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 80 theme receptor4 592:600 arg1 receptor 615:622 a Toll-like receptor4 (TLR4) decoy receptor 580:622 a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules 580:668 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 80 theme receptor4 592:600 arg1 protein 571:577 a model repeat protein 556:577 a model repeat protein 556:577 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 8 81 theme dynamics 1445:1452 arg1 simulations 1454:1464 Molecular dynamics simulations 1435:1464 Molecular dynamics simulations 1435:1464 Molecular dynamics simulations and energetic analysis indicate that an additive effect by two mutations occurring at nearby modules was the major contributor to the remarkable increase in the binding affinities. 22363519 4 82 theme differentiation 781:795 arg1 protein 797:803 myeloid differentiation protein 2 773:805 myeloid differentiation protein 2 (MD2) 773:811 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 4 82 theme differentiation 781:795 arg1 MD2 808:810 MD2 808:810 MD2 808:810 As a model repeat protein, a Toll-like receptor4 (TLR4) decoy receptor composed of leucine-rich repeat (LRR) modules was used, and its interaction interface was rationally engineered to increase the binding affinity for myeloid differentiation protein 2 (MD2). 22363519 5 83 theme decoy 860:864 arg1 receptor 866:873 the decoy receptor 856:873 the decoy receptor 856:873 Based on the complex crystal structure of the decoy receptor with MD2, we first designed single amino acid substitutions in the decoy receptor, and obtained three variants showing a binding affinity (K(D)) one-order of magnitude higher than the wild-type decoy receptor. 15003450 0 0 theme alpha-galactosidase 66:84 arg1 structure 47:55 structure 47:55 structure of human alpha-galactosidase 47:84 The molecular defect leading to Fabry disease: structure of human alpha-galactosidase. 15003450 7 1 dep nucleophile 1102:1112 arg1 a 1100:1100 a 1100:1100 a 1100:1100 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 1 2 with males 165:169 arg1 impairment 221:230 cardiac impairment 213:230 cardiac impairment 213:230 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 2 with males 165:169 arg1 symptoms 243:250 other symptoms 237:250 other symptoms 237:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 2 with males 165:169 arg1 pain 184:187 chronic pain 176:187 chronic pain 176:187 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 2 with males 165:169 arg1 degeneration 199:210 vascular degeneration 190:210 vascular degeneration 190:210 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 5 3 from sites 687:691 arg1 dimer 713:717 the glycoprotein dimer 696:717 the glycoprotein dimer 696:717 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 0 4 theme human 60:64 arg1 alpha-galactosidase 66:84 human alpha-galactosidase 60:84 human alpha-galactosidase 60:84 The molecular defect leading to Fabry disease: structure of human alpha-galactosidase. 15003450 3 5 theme X-ray 492:496 arg1 crystallography 498:512 X-ray crystallography 492:512 X-ray crystallography 492:512 Here, we present the structure of the human alpha-GAL glycoprotein determined by X-ray crystallography. 15003450 8 6 theme three-dimensional 1297:1313 arg1 structure 1315:1323 the three-dimensional structure 1293:1323 the three-dimensional structure 1293:1323 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 1 7 theme other 237:241 arg1 symptoms 243:250 other symptoms 237:250 other symptoms 237:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 6 8 from glycolipids 872:882 arg1 galactose 839:847 galactose 839:847 galactose from glycoproteins and glycolipids 839:882 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 2 9 theme lysosomal 271:279 arg1 alpha-galactosidase 288:306 the lysosomal enzyme alpha-galactosidase 267:306 the lysosomal enzyme alpha-galactosidase (alpha-GAL) 267:318 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 2 9 theme lysosomal 271:279 arg1 alpha-GAL 309:317 alpha-GAL 309:317 alpha-GAL 309:317 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 2 10 theme enzyme 281:286 arg1 alpha-galactosidase 288:306 the lysosomal enzyme alpha-galactosidase 267:306 the lysosomal enzyme alpha-galactosidase (alpha-GAL) 267:318 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 2 10 theme enzyme 281:286 arg1 alpha-GAL 309:317 alpha-GAL 309:317 alpha-GAL 309:317 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 4 11 theme beta/alpha 576:585 arg1 domain 589:594 a (beta/alpha)8 domain 573:594 a (beta/alpha)8 domain with the active site 573:615 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 5 12 theme glycoprotein 700:711 arg1 dimer 713:717 the glycoprotein dimer 696:717 the glycoprotein dimer 696:717 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 8 13 theme nonsense 1271:1278 arg1 mutations 1280:1288 245 missense and nonsense mutations 1254:1288 mutations 1280:1288 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 8 14 theme missense 1258:1265 arg1 locations 1241:1249 the locations 1237:1249 the locations of 245 missense and nonsense mutations in the three-dimensional structure 1237:1323 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 3 15 theme alpha-GAL 455:463 arg1 glycoprotein 465:476 the human alpha-GAL glycoprotein 445:476 the human alpha-GAL glycoprotein determined by X-ray crystallography 445:512 Here, we present the structure of the human alpha-GAL glycoprotein determined by X-ray crystallography. 15003450 8 16 from mutation 1157:1164 arg1 alpha-GAL 1169:1177 alpha-GAL 1169:1177 alpha-GAL 1169:1177 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 5 17 theme mannose-6-phosphate 772:790 arg1 receptor 792:799 the mannose-6-phosphate receptor 768:799 the mannose-6-phosphate receptor 768:799 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 8 18 theme mutations 1280:1288 arg1 locations 1241:1249 the locations 1237:1249 the locations of 245 missense and nonsense mutations in the three-dimensional structure 1237:1323 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 4 19 theme antiparallel 624:635 arg1 domain 642:647 an antiparallel beta domain 621:647 an antiparallel beta domain 621:647 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 9 20 theme Fabry 1366:1370 arg1 disease 1372:1378 Fabry disease 1366:1378 Fabry disease 1366:1378 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 5 21 theme lysosomal 744:752 arg1 transport 754:762 lysosomal transport 744:762 lysosomal transport via the mannose-6-phosphate receptor 744:799 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 4 22 with homodimer 534:542 arg1 monomer 554:560 each monomer 549:560 each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain 549:647 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 0 23 theme molecular 4:12 arg1 defect 14:19 The molecular defect 0:19 The molecular defect leading to Fabry disease 0:44 The molecular defect leading to Fabry disease: structure of human alpha-galactosidase. 15003450 7 24 theme acid 1056:1059 arg1 D231 1080:1083 D231 1080:1083 D231 1080:1083 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 24 theme acid 1056:1059 arg1 D170 1071:1074 D170 1071:1074 D170 1071:1074 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 24 theme acid 1056:1059 arg1 residues 1061:1068 two aspartic acid residues 1043:1068 two aspartic acid residues (D170 and D231) 1043:1084 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 24 theme acid 1056:1059 arg1 acid/base 1121:1129 an acid/base 1118:1129 an acid/base 1118:1129 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 5 25 gly glycoprotein 700:711 arg1 glycoprotein 700:711 the glycoprotein dimer 696:717 the glycoprotein dimer 696:717 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 5 26 theme N-linked 650:657 arg1 carbohydrate 659:670 N-linked carbohydrate 650:670 N-linked carbohydrate 650:670 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 9 27 theme molecular 1398:1406 arg1 diseases 1408:1415 molecular diseases 1398:1415 molecular diseases 1398:1415 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 6 28 theme catalytic 955:963 arg1 product 965:971 its catalytic product 951:971 its catalytic product 951:971 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 7 29 theme residues 1061:1068 arg1 location 1031:1038 the location 1027:1038 the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively 1027:1143 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 8 30 from locations 1241:1249 arg1 structure 1315:1323 the three-dimensional structure 1293:1323 the three-dimensional structure 1293:1323 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 3 31 gly glycoprotein 465:476 arg1 glycoprotein 465:476 the human alpha-GAL glycoprotein 445:476 the human alpha-GAL glycoprotein determined by X-ray crystallography 445:512 Here, we present the structure of the human alpha-GAL glycoprotein determined by X-ray crystallography. 15003450 1 32 theme chronic 176:182 arg1 pain 184:187 chronic pain 176:187 chronic pain 176:187 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 6 33 from glycoproteins 854:866 arg1 galactose 839:847 galactose 839:847 galactose from glycoproteins and glycolipids 839:882 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 8 34 theme point 1151:1155 arg1 mutation 1157:1164 a point mutation 1149:1164 a point mutation in alpha-GAL 1149:1177 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 6 35 gly glycoproteins 854:866 arg1 glycoproteins 854:866 glycoproteins 854:866 glycoproteins 854:866 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 4 36 theme active 605:610 arg1 site 612:615 the active site 601:615 the active site 601:615 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 1 37 theme Fabry 87:91 arg1 disease 93:99 Fabry disease 87:99 Fabry disease 87:99 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 37 theme Fabry 87:91 arg1 disease 134:140 an X-linked lysosomal storage disease 104:140 an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms 104:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 9 38 theme diseases 1408:1415 arg1 realm 1389:1393 the realm 1385:1393 the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients 1385:1535 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 7 39 theme aspartic 1047:1054 arg1 D231 1080:1083 D231 1080:1083 D231 1080:1083 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 39 theme aspartic 1047:1054 arg1 D170 1071:1074 D170 1071:1074 D170 1071:1074 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 39 theme aspartic 1047:1054 arg1 residues 1061:1068 two aspartic acid residues 1043:1068 two aspartic acid residues (D170 and D231) 1043:1084 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 39 theme aspartic 1047:1054 arg1 acid/base 1121:1129 an acid/base 1118:1129 an acid/base 1118:1129 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 4 40 with domain 642:647 arg1 site 612:615 the active site 601:615 the active site 601:615 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 0 41 dep structure 47:55 arg1 defect 14:19 The molecular defect 0:19 The molecular defect leading to Fabry disease 0:44 The molecular defect leading to Fabry disease: structure of human alpha-galactosidase. 15003450 8 42 theme Fabry 1191:1195 arg1 disease 1197:1203 Fabry disease 1191:1203 Fabry disease 1191:1203 As a point mutation in alpha-GAL can lead to Fabry disease, we have catalogued and plotted the locations of 245 missense and nonsense mutations in the three-dimensional structure. 15003450 0 43 theme Fabry 32:36 arg1 disease 38:44 Fabry disease 32:44 Fabry disease 32:44 The molecular defect leading to Fabry disease: structure of human alpha-galactosidase. 15003450 6 44 theme alpha-GAL 936:944 arg1 complex 925:931 the complex 921:931 the complex of alpha-GAL with its catalytic product 921:971 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 1 45 theme vascular 190:197 arg1 degeneration 199:210 vascular degeneration 190:210 vascular degeneration 190:210 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 4 46 theme beta 637:640 arg1 domain 642:647 an antiparallel beta domain 621:647 an antiparallel beta domain 621:647 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 2 47 theme disease 393:399 arg1 symptoms 401:408 Fabry disease symptoms 387:408 Fabry disease symptoms 387:408 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 7 48 dep residues 1061:1068 arg1 D231 1080:1083 D231 1080:1083 D231 1080:1083 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 48 dep residues 1061:1068 arg1 D170 1071:1074 D170 1071:1074 D170 1071:1074 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 7 48 dep residues 1061:1068 arg1 residues 1061:1068 two aspartic acid residues 1043:1068 two aspartic acid residues (D170 and D231) 1043:1084 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 9 49 theme alpha-GAL 1349:1357 arg1 structure 1330:1338 The structure 1326:1338 The structure of human alpha-GAL 1326:1357 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 4 50 with domain 589:594 arg1 site 612:615 the active site 601:615 the active site 601:615 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 9 51 theme human 1343:1347 arg1 alpha-GAL 1349:1357 human alpha-GAL 1343:1357 human alpha-GAL 1343:1357 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 2 52 theme Fabry 387:391 arg1 disease 393:399 Fabry disease 387:399 Fabry disease symptoms 387:408 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 6 53 theme complex 925:931 arg1 structure 908:916 the structure 904:916 the structure of the complex of alpha-GAL with its catalytic product 904:971 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 6 54 with complex 925:931 arg1 product 965:971 its catalytic product 951:971 its catalytic product 951:971 To understand how the enzyme cleaves galactose from glycoproteins and glycolipids, we also determined the structure of the complex of alpha-GAL with its catalytic product. 15003450 5 55 link N-linked 650:657 arg1 carbohydrate 659:670 N-linked carbohydrate 650:670 N-linked carbohydrate 650:670 N-linked carbohydrate appears at six sites in the glycoprotein dimer, revealing the basis for lysosomal transport via the mannose-6-phosphate receptor. 15003450 4 56 contain containing 562:571 arg2 domain 642:647 an antiparallel beta domain 621:647 an antiparallel beta domain 621:647 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 4 56 contain containing 562:571 arg2 domain 589:594 a (beta/alpha)8 domain 573:594 a (beta/alpha)8 domain with the active site 573:615 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 4 56 contain containing 562:571 arg1 monomer 554:560 each monomer 549:560 each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain 549:647 The structure is a homodimer with each monomer containing a (beta/alpha)8 domain with the active site and an antiparallel beta domain. 15003450 7 57 theme enzyme 1005:1010 arg1 mechanism 988:996 The catalytic mechanism 974:996 The catalytic mechanism of the enzyme 974:1010 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 3 58 theme glycoprotein 465:476 arg1 structure 432:440 the structure 428:440 the structure of the human alpha-GAL glycoprotein determined by X-ray crystallography 428:512 Here, we present the structure of the human alpha-GAL glycoprotein determined by X-ray crystallography. 15003450 3 59 theme human 449:453 arg1 glycoprotein 465:476 the human alpha-GAL glycoprotein 445:476 the human alpha-GAL glycoprotein determined by X-ray crystallography 445:512 Here, we present the structure of the human alpha-GAL glycoprotein determined by X-ray crystallography. 15003450 1 60 theme X-linked 107:114 arg1 disease 93:99 Fabry disease 87:99 Fabry disease 87:99 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 60 theme X-linked 107:114 arg1 disease 134:140 an X-linked lysosomal storage disease 104:140 an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms 104:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 9 61 theme patients 1528:1535 arg1 treatment 1515:1523 the clinical treatment 1502:1523 the clinical treatment of patients 1502:1535 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 1 62 theme lysosomal 116:124 arg1 disease 93:99 Fabry disease 87:99 Fabry disease 87:99 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 62 theme lysosomal 116:124 arg1 disease 134:140 an X-linked lysosomal storage disease 104:140 an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms 104:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 2 63 from Deficiency 253:262 arg1 alpha-galactosidase 288:306 the lysosomal enzyme alpha-galactosidase 267:306 the lysosomal enzyme alpha-galactosidase (alpha-GAL) 267:318 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 2 63 from Deficiency 253:262 arg1 alpha-GAL 309:317 alpha-GAL 309:317 alpha-GAL 309:317 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 15003450 1 64 link X-linked 107:114 arg1 disease 93:99 Fabry disease 87:99 Fabry disease 87:99 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 64 link X-linked 107:114 arg1 disease 134:140 an X-linked lysosomal storage disease 104:140 an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms 104:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 9 65 theme disease 1466:1472 arg1 phenotypes 1474:1483 the disease phenotypes 1462:1483 the disease phenotypes 1462:1483 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 1 66 theme storage 126:132 arg1 disease 93:99 Fabry disease 87:99 Fabry disease 87:99 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 66 theme storage 126:132 arg1 disease 134:140 an X-linked lysosomal storage disease 104:140 an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms 104:250 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 1 67 theme cardiac 213:219 arg1 impairment 221:230 cardiac impairment 213:230 cardiac impairment 213:230 Fabry disease is an X-linked lysosomal storage disease afflicting 1 in 40,000 males with chronic pain, vascular degeneration, cardiac impairment, and other symptoms. 15003450 7 68 theme catalytic 978:986 arg1 mechanism 988:996 The catalytic mechanism 974:996 The catalytic mechanism of the enzyme 974:1010 The catalytic mechanism of the enzyme is revealed by the location of two aspartic acid residues (D170 and D231), which act as a nucleophile and an acid/base, respectively. 15003450 9 69 theme structural 1442:1451 arg1 basis 1453:1457 the structural basis 1438:1457 the structural basis of the disease phenotypes 1438:1483 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 9 70 theme phenotypes 1474:1483 arg1 basis 1453:1457 the structural basis 1438:1457 the structural basis of the disease phenotypes 1438:1483 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 9 71 theme clinical 1506:1513 arg1 treatment 1515:1523 the clinical treatment 1502:1523 the clinical treatment of patients 1502:1535 The structure of human alpha-GAL brings Fabry disease into the realm of molecular diseases, where insights into the structural basis of the disease phenotypes might help guide the clinical treatment of patients. 15003450 2 72 theme substrate 350:358 arg1 accumulation 330:341 an accumulation 327:341 an accumulation 327:341 Deficiency in the lysosomal enzyme alpha-galactosidase (alpha-GAL) causes an accumulation of its substrate, which ultimately leads to Fabry disease symptoms. 25601083 0 0 theme -mediated 56:64 arg1 adenylylation/AMPylation 66:89 Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation 21:89 Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation 21:89 A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. 25601083 4 1 theme Huntingtin 669:678 arg1 HYPE 663:666 HYPE 663:666 HYPE (Huntingtin yeast interacting protein E) 663:707 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 1 theme Huntingtin 669:678 arg1 E 706:706 Huntingtin yeast interacting protein E 669:706 Huntingtin yeast interacting protein E 669:706 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 1 2 theme endoplasmic 145:155 arg1 ER 168:169 ER 168:169 ER 168:169 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 2 theme endoplasmic 145:155 arg1 reticulum 157:165 endoplasmic reticulum 145:165 endoplasmic reticulum (ER) homeostasis 145:182 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 10 3 theme UPR 1382:1384 arg1 regulator 1386:1394 a new UPR regulator 1376:1394 a new UPR regulator 1376:1394 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 10 3 theme UPR 1382:1384 arg1 HYPE 1368:1371 HYPE 1368:1371 HYPE 1368:1371 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 5 4 theme reticulum 854:862 arg1 lumen 829:833 the lumen 825:833 the lumen of the endoplasmic reticulum 825:862 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 1 5 theme reticulum 157:165 arg1 homeostasis 172:182 endoplasmic reticulum (ER) homeostasis 145:182 endoplasmic reticulum (ER) homeostasis 145:182 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 7 6 theme misfolded 1142:1150 arg1 proteins 1152:1159 misfolded proteins 1142:1159 misfolded proteins 1142:1159 We found that adenylylation enhances BiP's ATPase activity, which is required for refolding misfolded proteins while coping with ER stress. 25601083 7 7 with coping 1167:1172 arg1 stress 1182:1187 ER stress 1179:1187 ER stress 1179:1187 We found that adenylylation enhances BiP's ATPase activity, which is required for refolding misfolded proteins while coping with ER stress. 25601083 10 8 theme new 1378:1380 arg1 regulator 1386:1394 a new UPR regulator 1376:1394 a new UPR regulator 1376:1394 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 10 8 theme new 1378:1380 arg1 HYPE 1368:1371 HYPE 1368:1371 HYPE 1368:1371 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 5 9 theme endoplasmic 842:852 arg1 reticulum 854:862 the endoplasmic reticulum 838:862 the endoplasmic reticulum 838:862 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 5 10 theme molecular 919:927 arg1 BiP 940:942 the ER molecular chaperone, BiP, 912:943 BiP 940:942 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 5 11 theme chaperone 929:937 arg1 BiP 940:942 the ER molecular chaperone, BiP, 912:943 BiP 940:942 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 2 12 theme unknown 331:337 arg1 role 339:342 a previously unknown role 318:342 a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction 318:473 We have discovered a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction. 25601083 4 13 theme protein 698:704 arg1 HYPE 663:666 HYPE 663:666 HYPE (Huntingtin yeast interacting protein E) 663:707 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 13 theme protein 698:704 arg1 E 706:706 Huntingtin yeast interacting protein E 669:706 Huntingtin yeast interacting protein E 669:706 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 0 14 theme unfolded 99:106 arg1 response 116:123 the unfolded protein response 95:123 the unfolded protein response 95:123 A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. 25601083 9 15 theme siRNA-mediated 1261:1274 arg1 knockdown 1276:1284 siRNA-mediated knockdown 1261:1284 siRNA-mediated knockdown of HYPE 1261:1292 Furthermore, siRNA-mediated knockdown of HYPE prevents the induction of an unfolded protein response. 25601083 10 16 theme first 1412:1416 arg1 data 1429:1432 the first functional data 1408:1432 the first functional data for Fic-mediated adenylylation in mammalian signaling 1408:1486 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 8 17 theme HYPE 1203:1206 arg1 levels 1219:1224 HYPE expression levels 1203:1224 HYPE expression levels 1203:1224 Accordingly, HYPE expression levels increase upon stress. 25601083 4 18 theme physiological 756:768 arg1 s 777:777 unknown physiological target(s) 748:778 unknown physiological target(s) 748:778 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 3 19 dep domain 562:567 arg1 filamentation 531:543 filamentation 531:543 filamentation induced by cAMP 531:559 A family of enzymes, defined by the presence of a Fic (filamentation induced by cAMP) domain, catalyzes this adenylylation reaction. 25601083 3 20 theme Fic 526:528 arg1 domain 562:567 a Fic (filamentation induced by cAMP) domain 524:567 a Fic (filamentation induced by cAMP) domain 524:567 A family of enzymes, defined by the presence of a Fic (filamentation induced by cAMP) domain, catalyzes this adenylylation reaction. 25601083 6 21 theme ER 1034:1035 arg1 sentinel 988:995 a sentinel 986:995 a sentinel for protein misfolding 986:1018 BiP functions as a sentinel for protein misfolding and maintains ER homeostasis. 25601083 6 21 theme ER 1034:1035 arg1 BiP 969:971 BiP 969:971 BiP 969:971 BiP functions as a sentinel for protein misfolding and maintains ER homeostasis. 25601083 6 21 theme ER 1034:1035 arg1 homeostasis 1037:1047 ER homeostasis 1034:1047 ER homeostasis 1034:1047 BiP functions as a sentinel for protein misfolding and maintains ER homeostasis. 25601083 2 22 theme modification 368:379 arg1 role 339:342 a previously unknown role 318:342 a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction 318:473 We have discovered a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction. 25601083 0 23 theme novel 2:6 arg1 link 8:11 A novel link 0:11 A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. 0:124 A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. 25601083 9 24 theme HYPE 1289:1292 arg1 knockdown 1276:1284 siRNA-mediated knockdown 1261:1284 siRNA-mediated knockdown of HYPE 1261:1292 Furthermore, siRNA-mediated knockdown of HYPE prevents the induction of an unfolded protein response. 25601083 0 25 dep -mediated 56:64 arg1 filamentation 26:38 filamentation 26:38 filamentation induced by cAMP 26:54 A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. 25601083 2 26 theme post-translational 349:366 arg1 modification 368:379 a post-translational modification 347:379 a post-translational modification termed adenylylation/AMPylation 347:411 We have discovered a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction. 25601083 1 27 theme functioning 254:264 arg1 UPR 293:295 UPR 293:295 UPR 293:295 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 27 theme functioning 254:264 arg1 response 283:290 a properly functioning unfolded protein response 243:290 a properly functioning unfolded protein response (UPR) 243:296 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 28 theme homeostasis 172:182 arg1 maintenance 130:140 The maintenance 126:140 The maintenance of endoplasmic reticulum (ER) homeostasis 126:182 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 28 theme homeostasis 172:182 arg1 aspect 198:203 a critical aspect 187:203 a critical aspect of determining cell fate 187:228 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 29 theme unfolded 266:273 arg1 UPR 293:295 UPR 293:295 UPR 293:295 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 29 theme unfolded 266:273 arg1 response 283:290 a properly functioning unfolded protein response 243:290 a properly functioning unfolded protein response (UPR) 243:296 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 10 30 theme Fic-mediated 1438:1449 arg1 adenylylation 1451:1463 Fic-mediated adenylylation 1438:1463 Fic-mediated adenylylation 1438:1463 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 4 31 theme adenylyltransferase 715:733 arg1 activity 735:742 adenylyltransferase activity 715:742 adenylyltransferase activity 715:742 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 2 32 theme signal 427:432 arg1 events 447:452 signal transduction events 427:452 signal transduction events 427:452 We have discovered a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction. 25601083 2 33 theme transduction 434:445 arg1 events 447:452 signal transduction events 427:452 signal transduction events 427:452 We have discovered a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction. 25601083 1 34 theme protein 275:281 arg1 UPR 293:295 UPR 293:295 UPR 293:295 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 34 theme protein 275:281 arg1 response 283:290 a properly functioning unfolded protein response 243:290 a properly functioning unfolded protein response (UPR) 243:296 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 0 35 theme protein 108:114 arg1 response 116:123 the unfolded protein response 95:123 the unfolded protein response 95:123 A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. 25601083 5 36 theme ER 916:917 arg1 BiP 940:942 the ER molecular chaperone, BiP, 912:943 BiP 940:942 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 4 37 theme human 613:617 arg1 protein 647:653 a single Fic protein 634:653 a single Fic protein 634:653 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 37 theme human 613:617 arg1 genome 619:624 The human genome 609:624 The human genome 609:624 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 7 38 theme ATPase 1093:1098 arg1 activity 1100:1107 BiP's ATPase activity 1087:1107 BiP's ATPase activity 1087:1107 We found that adenylylation enhances BiP's ATPase activity, which is required for refolding misfolded proteins while coping with ER stress. 25601083 5 39 theme hydrophobic 872:882 arg1 terminus 886:893 its hydrophobic N terminus 868:893 its hydrophobic N terminus 868:893 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 4 40 with protein 647:653 arg1 s 777:777 unknown physiological target(s) 748:778 unknown physiological target(s) 748:778 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 40 with protein 647:653 arg1 activity 735:742 adenylyltransferase activity 715:742 adenylyltransferase activity 715:742 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 6 41 theme protein 1001:1007 arg1 misfolding 1009:1018 protein misfolding 1001:1018 protein misfolding 1001:1018 BiP functions as a sentinel for protein misfolding and maintains ER homeostasis. 25601083 1 42 theme critical 189:196 arg1 maintenance 130:140 The maintenance 126:140 The maintenance of endoplasmic reticulum (ER) homeostasis 126:182 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 1 42 theme critical 189:196 arg1 aspect 198:203 a critical aspect 187:203 a critical aspect of determining cell fate 187:228 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 4 43 theme Fic 643:645 arg1 protein 647:653 a single Fic protein 634:653 a single Fic protein 634:653 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 43 theme Fic 643:645 arg1 genome 619:624 The human genome 609:624 The human genome 609:624 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 10 44 theme functional 1418:1427 arg1 data 1429:1432 the first functional data 1408:1432 the first functional data for Fic-mediated adenylylation in mammalian signaling 1408:1486 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 3 45 theme domain 562:567 arg1 presence 512:519 the presence 508:519 the presence of a Fic (filamentation induced by cAMP) domain 508:567 A family of enzymes, defined by the presence of a Fic (filamentation induced by cAMP) domain, catalyzes this adenylylation reaction. 25601083 4 46 theme interacting 686:696 arg1 HYPE 663:666 HYPE 663:666 HYPE (Huntingtin yeast interacting protein E) 663:707 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 46 theme interacting 686:696 arg1 E 706:706 Huntingtin yeast interacting protein E 669:706 Huntingtin yeast interacting protein E 669:706 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 9 47 theme unfolded 1323:1330 arg1 response 1340:1347 an unfolded protein response 1320:1347 an unfolded protein response 1320:1347 Furthermore, siRNA-mediated knockdown of HYPE prevents the induction of an unfolded protein response. 25601083 10 48 theme mammalian 1468:1476 arg1 signaling 1478:1486 mammalian signaling 1468:1486 mammalian signaling 1468:1486 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 4 49 theme yeast 680:684 arg1 HYPE 663:666 HYPE 663:666 HYPE (Huntingtin yeast interacting protein E) 663:707 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 49 theme yeast 680:684 arg1 E 706:706 Huntingtin yeast interacting protein E 669:706 Huntingtin yeast interacting protein E 669:706 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 9 50 theme protein 1332:1338 arg1 response 1340:1347 an unfolded protein response 1320:1347 an unfolded protein response 1320:1347 Furthermore, siRNA-mediated knockdown of HYPE prevents the induction of an unfolded protein response. 25601083 10 51 from data 1429:1432 arg1 signaling 1478:1486 mammalian signaling 1468:1486 mammalian signaling 1468:1486 Thus, we identify HYPE as a new UPR regulator and provide the first functional data for Fic-mediated adenylylation in mammalian signaling. 25601083 5 52 theme N 884:884 arg1 terminus 886:893 its hydrophobic N terminus 868:893 its hydrophobic N terminus 868:893 Here, we demonstrate that HYPE localizes to the lumen of the endoplasmic reticulum via its hydrophobic N terminus and adenylylates the ER molecular chaperone, BiP, at Ser-365 and Thr-366. 25601083 9 53 theme response 1340:1347 arg1 induction 1307:1315 the induction 1303:1315 the induction of an unfolded protein response 1303:1347 Furthermore, siRNA-mediated knockdown of HYPE prevents the induction of an unfolded protein response. 25601083 4 54 theme unknown 748:754 arg1 s 777:777 unknown physiological target(s) 748:778 unknown physiological target(s) 748:778 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 55 theme target 770:775 arg1 s 777:777 unknown physiological target(s) 748:778 unknown physiological target(s) 748:778 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 3 56 theme enzymes 488:494 arg1 family 478:483 A family 476:483 A family of enzymes 476:494 A family of enzymes, defined by the presence of a Fic (filamentation induced by cAMP) domain, catalyzes this adenylylation reaction. 25601083 3 57 theme adenylylation 585:597 arg1 reaction 599:606 this adenylylation reaction 580:606 this adenylylation reaction 580:606 A family of enzymes, defined by the presence of a Fic (filamentation induced by cAMP) domain, catalyzes this adenylylation reaction. 25601083 8 58 theme expression 1208:1217 arg1 levels 1219:1224 HYPE expression levels 1203:1224 HYPE expression levels 1203:1224 Accordingly, HYPE expression levels increase upon stress. 25601083 4 59 theme single 636:641 arg1 protein 647:653 a single Fic protein 634:653 a single Fic protein 634:653 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 4 59 theme single 636:641 arg1 genome 619:624 The human genome 609:624 The human genome 609:624 The human genome encodes a single Fic protein, called HYPE (Huntingtin yeast interacting protein E), with adenylyltransferase activity but unknown physiological target(s). 25601083 1 60 theme cell 220:223 arg1 fate 225:228 cell fate 220:228 cell fate 220:228 The maintenance of endoplasmic reticulum (ER) homeostasis is a critical aspect of determining cell fate and requires a properly functioning unfolded protein response (UPR). 25601083 7 61 theme ER 1179:1180 arg1 stress 1182:1187 ER stress 1179:1187 ER stress 1179:1187 We found that adenylylation enhances BiP's ATPase activity, which is required for refolding misfolded proteins while coping with ER stress. 25601083 2 62 theme UPR 461:463 arg1 induction 465:473 UPR induction 461:473 UPR induction 461:473 We have discovered a previously unknown role of a post-translational modification termed adenylylation/AMPylation in regulating signal transduction events during UPR induction. 11406581 7 0 theme novel 1065:1069 arg1 mechanism 1071:1079 a novel mechanism 1063:1079 a novel mechanism which may contribute to S.pyogenes pathogenesis 1063:1127 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 2 1 contain has 326:328 arg1 S.pyogenes 315:324 S.pyogenes 315:324 S.pyogenes 315:324 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 2 1 contain has 326:328 arg2 ability 334:340 the ability to hydrolyze the chitobiose core of the asparagine-linked glycan 330:405 the ability to hydrolyze the chitobiose core of the asparagine-linked glycan 330:405 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 7 2 theme human 1040:1044 arg1 IgG 1046:1048 human IgG 1040:1048 human IgG 1040:1048 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 6 3 theme papain-like 911:921 arg1 manner 923:928 a papain-like manner 909:928 a papain-like manner 909:928 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 0 4 theme endoglycosidase 65:79 arg1 activity 81:88 endoglycosidase activity 65:88 endoglycosidase activity on human IgG 65:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 0 5 theme human 93:97 arg1 IgG 99:101 human IgG 93:101 human IgG 93:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 4 6 theme soluble 627:633 arg1 IgG 635:637 purified soluble IgG 618:637 purified soluble IgG as well as IgG bound to the bacterial surface 618:683 EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. 11406581 1 7 theme complement 267:276 arg1 factors 278:284 complement factors 267:284 complement factors 267:284 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 2 8 theme plasma 483:488 arg1 presence 465:472 the presence 461:472 the presence of human plasma 461:488 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 2 9 theme immuno 410:415 arg1 IgG 429:431 IgG 429:431 IgG 429:431 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 2 9 theme immuno 410:415 arg1 G 426:426 immuno globulin G 410:426 immuno globulin G (IgG) 410:432 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 2 10 theme asparagine-linked 382:398 arg1 glycan 400:405 the asparagine-linked glycan 378:405 the asparagine-linked glycan 378:405 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 2 11 theme human 477:481 arg1 plasma 483:488 human plasma 477:488 human plasma 477:488 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 4 12 theme purified 618:625 arg1 IgG 635:637 purified soluble IgG 618:637 purified soluble IgG as well as IgG bound to the bacterial surface 618:683 EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. 11406581 5 13 theme isogenic 735:742 arg1 mutant 750:755 an isogenic EndoS mutant 732:755 an isogenic EndoS mutant 732:755 EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG. 11406581 5 14 theme EndoS 744:748 arg1 mutant 750:755 an isogenic EndoS mutant 732:755 an isogenic EndoS mutant 732:755 EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG. 11406581 6 15 theme hinge 893:897 arg1 region 899:904 the hinge region 889:904 the hinge region 889:904 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 7 16 theme bacterial 993:1001 arg1 pathogen 1003:1010 a bacterial pathogen 991:1010 a bacterial pathogen that selectively hydrolyzes human IgG 991:1048 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 0 17 theme novel 9:13 arg1 protein 24:30 a novel secreted protein 7:30 a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG 7:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 0 17 theme novel 9:13 arg1 EndoS 0:4 EndoS 0:4 EndoS 0:4 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 2 18 theme globulin 417:424 arg1 IgG 429:431 IgG 429:431 IgG 429:431 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 2 18 theme globulin 417:424 arg1 G 426:426 immuno globulin G 410:426 immuno globulin G (IgG) 410:432 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 6 19 from IgG 882:884 arg1 region 899:904 the hinge region 889:904 the hinge region 889:904 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 0 20 from activity 81:88 arg1 IgG 99:101 human IgG 93:101 human IgG 93:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 6 21 theme secreted 826:833 arg1 SpeB 869:872 the secreted streptococcal cysteine proteinase SpeB 822:872 the secreted streptococcal cysteine proteinase SpeB 822:872 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 1 22 theme important 133:141 arg1 pathogen 149:156 an important human pathogen 130:156 an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors 130:284 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 1 22 theme important 133:141 arg1 pyogenes 118:125 Streptococcus pyogenes 104:125 Streptococcus pyogenes 104:125 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 3 23 theme kDa 553:555 arg1 protein 557:563 a novel 108 kDa protein 541:563 a novel 108 kDa protein denoted EndoS 541:577 This activity is associated with the secretion of a novel 108 kDa protein denoted EndoS. 11406581 0 24 theme secreted 15:22 arg1 protein 24:30 a novel secreted protein 7:30 a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG 7:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 0 24 theme secreted 15:22 arg1 EndoS 0:4 EndoS 0:4 EndoS 0:4 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 7 25 theme endoglycosidase 963:977 arg1 example 949:955 the first example 939:955 the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG 939:1048 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 7 25 theme endoglycosidase 963:977 arg1 This 931:934 This 931:934 This 931:934 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 2 26 link asparagine-linked 382:398 arg1 glycan 400:405 the asparagine-linked glycan 378:405 the asparagine-linked glycan 378:405 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 3 27 theme protein 557:563 arg1 secretion 528:536 the secretion 524:536 the secretion of a novel 108 kDa protein denoted EndoS 524:577 This activity is associated with the secretion of a novel 108 kDa protein denoted EndoS. 11406581 2 28 theme glycan 400:405 arg1 core 370:373 the chitobiose core 355:373 the chitobiose core of the asparagine-linked glycan 355:405 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 0 29 theme Streptococcus 37:49 arg1 pyogenes 51:58 Streptococcus pyogenes 37:58 Streptococcus pyogenes 37:58 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 7 30 theme S.pyogenes 1105:1114 arg1 pathogenesis 1116:1127 S.pyogenes pathogenesis 1105:1127 S.pyogenes pathogenesis 1105:1127 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 0 31 from pyogenes 51:58 arg1 protein 24:30 a novel secreted protein 7:30 a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG 7:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 0 31 from pyogenes 51:58 arg1 EndoS 0:4 EndoS 0:4 EndoS 0:4 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 3 32 theme novel 543:547 arg1 protein 557:563 a novel 108 kDa protein 541:563 a novel 108 kDa protein denoted EndoS 541:577 This activity is associated with the secretion of a novel 108 kDa protein denoted EndoS. 11406581 5 33 from glycan 781:786 arg1 IgG 791:793 IgG 791:793 IgG 791:793 EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG. 11406581 1 34 theme Streptococcus 104:116 arg1 pathogen 149:156 an important human pathogen 130:156 an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors 130:284 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 1 34 theme Streptococcus 104:116 arg1 pyogenes 118:125 Streptococcus pyogenes 104:125 Streptococcus pyogenes 104:125 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 6 35 theme proteinase 858:867 arg1 SpeB 869:872 the secreted streptococcal cysteine proteinase SpeB 822:872 the secreted streptococcal cysteine proteinase SpeB 822:872 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 5 36 from activity 712:719 arg1 IgG 724:726 IgG 724:726 IgG 724:726 EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG. 11406581 4 37 contain has 586:588 arg2 activity 606:613 endoglycosidase activity 590:613 endoglycosidase activity 590:613 EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. 11406581 4 37 contain has 586:588 arg1 EndoS 580:584 EndoS 580:584 EndoS 580:584 EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. 11406581 6 38 theme cysteine 849:856 arg1 proteinase 858:867 streptococcal cysteine proteinase 835:867 the secreted streptococcal cysteine proteinase SpeB 822:872 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 1 39 theme defense 223:229 arg1 system 231:236 the humoral defense system 211:236 the humoral defense system 211:236 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 7 40 theme first 943:947 arg1 example 949:955 the first example 939:955 the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG 939:1048 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 7 40 theme first 943:947 arg1 This 931:934 This 931:934 This 931:934 This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. 11406581 4 41 theme bacterial 667:675 arg1 surface 677:683 the bacterial surface 663:683 the bacterial surface 663:683 EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. 11406581 1 42 theme humoral 215:221 arg1 system 231:236 the humoral defense system 211:236 the humoral defense system 211:236 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 4 43 theme endoglycosidase 590:604 arg1 activity 606:613 endoglycosidase activity 590:613 endoglycosidase activity 590:613 EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. 11406581 2 44 theme chitobiose 359:368 arg1 core 370:373 the chitobiose core 355:373 the chitobiose core of the asparagine-linked glycan 355:405 In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. 11406581 6 45 theme streptococcal 835:847 arg1 proteinase 858:867 streptococcal cysteine proteinase 835:867 the secreted streptococcal cysteine proteinase SpeB 822:872 In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. 11406581 0 46 with protein 24:30 arg1 activity 81:88 endoglycosidase activity 65:88 endoglycosidase activity on human IgG 65:101 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. 11406581 1 47 theme human 143:147 arg1 pathogen 149:156 an important human pathogen 130:156 an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors 130:284 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 11406581 1 47 theme human 143:147 arg1 pyogenes 118:125 Streptococcus pyogenes 104:125 Streptococcus pyogenes 104:125 Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. 20357243 7 0 theme G2a 1255:1257 arg1 autoantibody 1270:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 7 1 from Development 1182:1192 arg1 mice 1207:1210 mice 1207:1210 mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver 1207:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 5 2 theme anti-D-sensitized 923:939 arg1 RBCs 941:944 anti-D-sensitized RBCs 923:944 anti-D-sensitized RBCs 923:944 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 7 3 theme monoclonal 1259:1268 arg1 autoantibody 1270:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 4 4 with pretreatment 734:745 arg1 EndoS 762:766 EndoS 762:766 EndoS 762:766 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 6 5 from Agglutination 1045:1057 arg1 blood 1125:1129 whole human blood 1113:1129 whole human blood 1113:1129 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 8 6 dep potential 1420:1428 arg1 therapeutic 1430:1440 therapeutic 1430:1440 therapeutic 1430:1440 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 8 7 theme potential 1420:1428 arg1 agent 1442:1446 a potential therapeutic agent 1418:1446 a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs 1418:1564 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 8 7 theme potential 1420:1428 arg1 EndoS 1409:1413 EndoS 1409:1413 EndoS 1409:1413 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 8 7 theme potential 1420:1428 arg1 alternative 1478:1488 an alternative 1475:1488 an alternative to current treatment regimens against antibody-mediated destruction of RBCs 1475:1564 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 4 8 contain containing 848:857 arg1 blood 842:846 blood 842:846 blood containing sensitized RBCs 842:873 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 4 8 contain containing 848:857 arg2 RBCs 870:873 sensitized RBCs 859:873 sensitized RBCs 859:873 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 7 9 theme anti-RBC 1231:1238 arg1 autoantibody 1270:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 7 10 from anemia 1197:1202 arg1 mice 1207:1210 mice 1207:1210 mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver 1207:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 7 11 from activation 1298:1307 arg1 liver 1357:1361 the liver 1353:1361 the liver 1353:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 2 12 theme blood 349:353 arg1 RBC 361:363 RBC 361:363 RBC 361:363 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 2 12 theme blood 349:353 arg1 cell 355:358 red blood cell 345:358 antibody-mediated red blood cell (RBC) destruction 327:376 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 5 13 theme oxygen 990:995 arg1 production 1008:1017 oxygen metabolite production 990:1017 oxygen metabolite production 990:1017 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 5 14 theme interleukin-8 962:974 arg1 secretion 976:984 interleukin-8 secretion 962:984 interleukin-8 secretion 962:984 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 3 15 theme phagocytosis 586:597 arg1 destruction 562:572 destruction 562:572 destruction of RBC, ie, phagocytosis, complement activation, and hemolysis 562:635 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 2 16 theme red 345:347 arg1 RBC 361:363 RBC 361:363 RBC 361:363 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 2 16 theme red 345:347 arg1 cell 355:358 red blood cell 345:358 antibody-mediated red blood cell (RBC) destruction 327:376 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 7 17 from mice 1207:1210 arg1 Development 1182:1192 Development 1182:1192 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver 1182:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 7 18 from erythrophagocytosis 1313:1331 arg1 liver 1357:1361 the liver 1353:1361 the liver 1353:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 2 19 theme antibody-mediated 327:343 arg1 destruction 366:376 antibody-mediated red blood cell (RBC) destruction 327:376 antibody-mediated red blood cell (RBC) destruction 327:376 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 6 20 theme whole 1113:1117 arg1 blood 1125:1129 whole human blood 1113:1129 whole human blood 1113:1129 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 2 21 theme adverse 452:458 arg1 effects 460:466 frequent adverse effects 443:466 frequent adverse effects 443:466 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 7 22 theme murine 1224:1229 arg1 autoantibody 1270:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 4 23 theme sensitized 859:868 arg1 RBCs 870:873 sensitized RBCs 859:873 sensitized RBCs 859:873 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 6 24 theme RBCs 1062:1065 arg1 Agglutination 1045:1057 Agglutination 1045:1057 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs 1045:1156 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 2 25 theme frequent 443:450 arg1 effects 460:466 frequent adverse effects 443:466 frequent adverse effects 443:466 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 0 26 theme IgG-specific 4:15 arg1 EndoS 33:37 The IgG-specific endoglycosidase EndoS 0:37 The IgG-specific endoglycosidase EndoS 0:37 The IgG-specific endoglycosidase EndoS inhibits both cellular and complement-mediated autoimmune hemolysis. 20357243 3 27 theme complement 600:609 arg1 activation 611:620 complement activation 600:620 complement activation 600:620 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 1 28 theme immunoglobulin 220:233 arg1 G 235:235 human immunoglobulin G 214:235 human immunoglobulin G 214:235 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 3 29 theme activation 611:620 arg1 destruction 562:572 destruction 562:572 destruction of RBC, ie, phagocytosis, complement activation, and hemolysis 562:635 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 4 30 theme RBCs 792:795 arg1 sensitization 775:787 sensitization 775:787 sensitization of RBCs 775:795 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 5 31 theme monocytes 897:905 arg1 effects 886:892 The toxic effects 876:892 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, 876:1018 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 6 32 theme human 1119:1123 arg1 blood 1125:1129 whole human blood 1113:1129 whole human blood 1113:1129 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 1 33 theme Streptococcus 119:131 arg1 pyogenes 133:140 Streptococcus pyogenes 119:140 Streptococcus pyogenes 119:140 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 3 34 theme hemolysis 627:635 arg1 destruction 562:572 destruction 562:572 destruction of RBC, ie, phagocytosis, complement activation, and hemolysis 562:635 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 5 35 theme metabolite 997:1006 arg1 production 1008:1017 oxygen metabolite production 990:1017 oxygen metabolite production 990:1017 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 0 36 theme endoglycosidase 17:31 arg1 EndoS 33:37 The IgG-specific endoglycosidase EndoS 0:37 The IgG-specific endoglycosidase EndoS 0:37 The IgG-specific endoglycosidase EndoS inhibits both cellular and complement-mediated autoimmune hemolysis. 20357243 3 37 theme RBC 577:579 arg1 destruction 562:572 destruction 562:572 destruction of RBC, ie, phagocytosis, complement activation, and hemolysis 562:635 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 8 38 theme current 1493:1499 arg1 regimens 1511:1518 current treatment regimens 1493:1518 current treatment regimens against antibody-mediated destruction of RBCs 1493:1564 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 2 39 theme cell 355:358 arg1 destruction 366:376 antibody-mediated red blood cell (RBC) destruction 327:376 antibody-mediated red blood cell (RBC) destruction 327:376 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 3 40 theme rabbit 509:514 arg1 anti-RBC 543:550 anti-RBC 543:550 anti-RBC 543:550 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 3 40 theme rabbit 509:514 arg1 antibodies 531:540 rabbit anti-human-RBC antibodies 509:540 rabbit anti-human-RBC antibodies (anti-RBC) 509:551 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 2 41 theme Autoimmune 286:295 arg1 anemia 307:312 Autoimmune hemolytic anemia 286:312 Autoimmune hemolytic anemia 286:312 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 6 42 theme hemolysis 1091:1099 arg1 Agglutination 1045:1057 Agglutination 1045:1057 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs 1045:1156 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 5 43 theme toxic 880:884 arg1 effects 886:892 The toxic effects 876:892 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, 876:1018 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 7 44 theme anemia 1197:1202 arg1 Development 1182:1192 Development 1182:1192 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver 1182:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 4 45 theme EndoS 833:837 arg1 addition 821:828 direct addition 814:828 direct addition of EndoS to blood containing sensitized RBCs 814:873 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 1 46 theme antibody 257:264 arg1 functions 275:283 antibody effector functions 257:283 antibody effector functions 257:283 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 8 47 theme treatment 1501:1509 arg1 regimens 1511:1518 current treatment regimens 1493:1518 current treatment regimens against antibody-mediated destruction of RBCs 1493:1564 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 3 48 dep anti-RhD 487:494 arg1 anti-D 497:502 anti-D 497:502 anti-D 497:502 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 5 49 dep RBCs 941:944 arg1 measured 950:957 measured 950:957 measured by interleukin-8 secretion and oxygen metabolite production 950:1017 The toxic effects of monocytes stimulated with anti-D-sensitized RBCs, as measured by interleukin-8 secretion and oxygen metabolite production, was restrained by EndoS. 20357243 0 50 theme cellular 53:60 arg1 hemolysis 97:105 both cellular and complement-mediated autoimmune hemolysis 48:105 both cellular and complement-mediated autoimmune hemolysis 48:105 The IgG-specific endoglycosidase EndoS inhibits both cellular and complement-mediated autoimmune hemolysis. 20357243 8 51 theme antibody-mediated 1528:1544 arg1 destruction 1546:1556 antibody-mediated destruction 1528:1556 antibody-mediated destruction of RBCs 1528:1564 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 4 52 theme anti-D 750:755 arg1 pretreatment 734:745 pretreatment 734:745 pretreatment of anti-D with EndoS before sensitization of RBCs 734:795 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 2 53 with treatment 396:404 arg1 corticosteroids 411:425 corticosteroids 411:425 corticosteroids that also cause frequent adverse effects 411:466 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 4 54 theme direct 814:819 arg1 addition 821:828 direct addition 814:828 direct addition of EndoS to blood containing sensitized RBCs 814:873 Phagocytosis by monocytes in vitro was inhibited by pretreatment of anti-D with EndoS before sensitization of RBCs and abrogated by direct addition of EndoS to blood containing sensitized RBCs. 20357243 1 55 theme immunomodulating 148:163 arg1 enzyme 165:170 an immunomodulating enzyme 145:170 an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions 145:283 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 1 55 theme immunomodulating 148:163 arg1 EndoS 108:112 EndoS 108:112 EndoS from Streptococcus pyogenes 108:140 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 1 56 from pyogenes 133:140 arg1 enzyme 165:170 an immunomodulating enzyme 145:170 an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions 145:283 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 1 56 from pyogenes 133:140 arg1 EndoS 108:112 EndoS 108:112 EndoS from Streptococcus pyogenes 108:140 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 1 57 theme effector 266:273 arg1 functions 275:283 antibody effector functions 257:283 antibody effector functions 257:283 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 7 58 from autoantibody 1270:1281 arg1 liver 1357:1361 the liver 1353:1361 the liver 1353:1361 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 3 59 theme ie 582:583 arg1 destruction 562:572 destruction 562:572 destruction of RBC, ie, phagocytosis, complement activation, and hemolysis 562:635 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 8 60 theme RBCs 1561:1564 arg1 destruction 1546:1556 antibody-mediated destruction 1528:1556 antibody-mediated destruction of RBCs 1528:1564 Our data indicate that EndoS is a potential therapeutic agent that might be evaluated as an alternative to current treatment regimens against antibody-mediated destruction of RBCs. 20357243 7 61 theme Kupffer 1336:1342 arg1 cells 1344:1348 Kupffer cells 1336:1348 Kupffer cells 1336:1348 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 3 62 theme anti-human-RBC 516:529 arg1 anti-RBC 543:550 anti-RBC 543:550 anti-RBC 543:550 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 3 62 theme anti-human-RBC 516:529 arg1 antibodies 531:540 rabbit anti-human-RBC antibodies 509:540 rabbit anti-human-RBC antibodies (anti-RBC) 509:551 We show here that anti-RhD (anti-D) and rabbit anti-human-RBC antibodies (anti-RBC) mediated destruction of RBC, ie, phagocytosis, complement activation, and hemolysis in vitro and in vivo was inhibited by EndoS. 20357243 0 63 theme autoimmune 86:95 arg1 hemolysis 97:105 both cellular and complement-mediated autoimmune hemolysis 48:105 both cellular and complement-mediated autoimmune hemolysis 48:105 The IgG-specific endoglycosidase EndoS inhibits both cellular and complement-mediated autoimmune hemolysis. 20357243 6 64 theme complement-mediated 1071:1089 arg1 hemolysis 1091:1099 complement-mediated hemolysis 1071:1099 complement-mediated hemolysis 1071:1099 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 1 65 from G 235:235 arg1 glycans 201:207 glycans 201:207 glycans from human immunoglobulin G 201:235 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 1 66 theme human 214:218 arg1 G 235:235 human immunoglobulin G 214:235 human immunoglobulin G 214:235 EndoS from Streptococcus pyogenes is an immunomodulating enzyme that specifically hydrolyzes glycans from human immunoglobulin G and thereby affects antibody effector functions. 20357243 0 67 theme complement-mediated 66:84 arg1 hemolysis 97:105 both cellular and complement-mediated autoimmune hemolysis 48:105 both cellular and complement-mediated autoimmune hemolysis 48:105 The IgG-specific endoglycosidase EndoS inhibits both cellular and complement-mediated autoimmune hemolysis. 20357243 2 68 theme hemolytic 297:305 arg1 anemia 307:312 Autoimmune hemolytic anemia 286:312 Autoimmune hemolytic anemia 286:312 Autoimmune hemolytic anemia is caused by antibody-mediated red blood cell (RBC) destruction and often resists treatment with corticosteroids that also cause frequent adverse effects. 20357243 6 69 theme rabbit 1141:1146 arg1 anti-RBCs 1148:1156 rabbit anti-RBCs 1141:1156 rabbit anti-RBCs 1141:1156 Agglutination of RBCs and complement-mediated hemolysis in vitro in whole human blood caused by rabbit anti-RBCs was inhibited by EndoS. 20357243 7 70 theme complement 1287:1296 arg1 activation 1298:1307 complement activation 1287:1307 complement activation 1287:1307 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 20357243 7 71 theme immunoglobulin 1240:1253 arg1 autoantibody 1270:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 a murine anti-RBC immunoglobulin G2a monoclonal autoantibody 1222:1281 Development of anemia in mice caused by a murine anti-RBC immunoglobulin G2a monoclonal autoantibody and complement activation and erythrophagocytosis by Kupffer cells in the liver were reduced by EndoS. 15965977 9 0 theme family 1657:1662 arg1 understanding 1627:1639 our understanding 1623:1639 our understanding of the rhomboid family of regulatory proteins 1623:1685 The characterization of this protein extends our understanding of the rhomboid family of regulatory proteins. 15965977 7 1 theme core 1406:1409 arg1 ectodomain 1411:1420 the EGF core ectodomain 1398:1420 the EGF core ectodomain 1398:1420 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 4 2 theme serine 834:839 arg1 rhomboid 716:723 an atypically long human rhomboid 691:723 an atypically long human rhomboid 691:723 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 4 2 theme serine 834:839 arg1 protease 841:848 a serine protease 832:848 a serine protease 832:848 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 1 3 theme Spitz 152:156 arg1 activity 114:121 The activity 110:121 The activity of the TGF-alpha-like ligand Spitz in Drosophila 110:170 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 8 4 theme functional 1557:1566 arg1 activity 1568:1575 functional activity 1557:1575 functional activity 1557:1575 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 6 5 with protein 1076:1082 arg1 extension 1119:1127 a long N-terminal cytoplasmic extension 1089:1127 a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface 1089:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 3 6 from number 492:497 arg1 vertebrates 528:538 vertebrates 528:538 vertebrates 528:538 The larger number of TGF-alpha-like ligands in vertebrates raises the possibility that they too might be subject to regulation by rhomboid-like proteins. 15965977 5 7 theme tissue 867:872 arg1 distribution 874:885 its tissue distribution 863:885 its tissue distribution 863:885 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 0 8 theme family 94:99 arg1 ligands 101:107 TGF-alpha family ligands 84:107 TGF-alpha family ligands 84:107 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 1 9 theme soluble 327:333 arg1 ligand 335:340 the soluble ligand 323:340 the soluble ligand 323:340 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 5 10 theme TGF-alpha-related 925:941 arg1 HB-EGF 951:956 the TGF-alpha-related protein HB-EGF 921:956 the TGF-alpha-related protein HB-EGF 921:956 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 8 11 from function 1449:1456 arg1 Drosophila 1482:1491 Drosophila 1482:1491 Drosophila 1482:1491 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 0 12 theme TGF-alpha 84:92 arg1 ligands 101:107 TGF-alpha family ligands 84:107 TGF-alpha family ligands 84:107 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 7 13 theme luminal 1373:1379 arg1 interaction 1381:1391 a luminal interaction 1371:1391 a luminal interaction with the EGF core ectodomain 1371:1420 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 5 14 theme protein 943:949 arg1 HB-EGF 951:956 the TGF-alpha-related protein HB-EGF 921:956 the TGF-alpha-related protein HB-EGF 921:956 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 4 15 theme rhomboid 716:723 arg1 cloning 655:661 cDNA cloning 650:661 cDNA cloning 650:661 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 4 15 theme rhomboid 716:723 arg1 sequence 679:686 polypeptide sequence 667:686 polypeptide sequence 667:686 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 6 16 theme endoplasmic 1198:1208 arg1 reticulum 1210:1218 the endoplasmic reticulum 1194:1218 the endoplasmic reticulum 1194:1218 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 4 17 theme critical 757:764 arg1 residues 766:773 critical residues 757:773 critical residues for serine protease activity 757:802 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 9 18 theme protein 1607:1613 arg1 characterization 1582:1597 The characterization 1578:1597 The characterization of this protein 1578:1613 The characterization of this protein extends our understanding of the rhomboid family of regulatory proteins. 15965977 0 19 theme homolog 37:43 arg1 Characterization 0:15 Characterization 0:15 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 0:108 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 6 20 theme cytoplasmic 1107:1117 arg1 extension 1119:1127 a long N-terminal cytoplasmic extension 1089:1127 a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface 1089:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 6 21 located found 1185:1189 arg2 extension 1119:1127 a long N-terminal cytoplasmic extension 1089:1127 a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface 1089:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 6 21 located found 1185:1189 arg1 reticulum 1210:1218 the endoplasmic reticulum 1194:1218 the endoplasmic reticulum 1194:1218 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 4 22 theme cDNA 650:653 arg1 cloning 655:661 cDNA cloning 650:661 cDNA cloning 650:661 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 9 23 theme regulatory 1667:1676 arg1 proteins 1678:1685 regulatory proteins 1667:1685 regulatory proteins 1667:1685 The characterization of this protein extends our understanding of the rhomboid family of regulatory proteins. 15965977 8 24 theme p100 1461:1464 arg1 /RHBDF1 1471:1477 p100(hRho)/RHBDF1 1461:1477 p100(hRho)/RHBDF1 1461:1477 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 6 25 theme N-terminal 1096:1105 arg1 extension 1119:1127 a long N-terminal cytoplasmic extension 1089:1127 a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface 1089:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 4 26 theme polypeptide 667:677 arg1 sequence 679:686 polypeptide sequence 667:686 polypeptide sequence 667:686 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 2 27 theme ligands 432:438 arg1 activation 469:478 consequent EGF receptor activation 445:478 consequent EGF receptor activation 445:478 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 2 27 theme ligands 432:438 arg1 processing 403:412 the processing 399:412 the processing of TGF-alpha-like ligands 399:438 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 7 28 theme TGF-alpha 1345:1353 arg1 ligands 1355:1361 TGF-alpha ligands 1345:1361 TGF-alpha ligands 1345:1361 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 2 29 from rhomboids 351:359 arg1 Drosophila 364:373 Drosophila 364:373 Drosophila 364:373 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 6 30 theme long 1091:1094 arg1 extension 1119:1127 a long N-terminal cytoplasmic extension 1089:1127 a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface 1089:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 0 31 theme human 22:26 arg1 homolog 37:43 a human rhomboid homolog 20:43 a human rhomboid homolog 20:43 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 0 31 theme human 22:26 arg1 p100hRho/RHBDF1 46:60 p100hRho/RHBDF1 46:60 p100hRho/RHBDF1 46:60 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 5 32 theme EGF/TGF-alpha 967:979 arg1 receptor 981:988 the EGF/TGF-alpha receptor 963:988 the EGF/TGF-alpha receptor 963:988 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 1 33 from activity 114:121 arg1 Drosophila 161:170 Drosophila 161:170 Drosophila 161:170 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 1 34 theme serine 272:277 arg1 protein 224:230 a seven-transmembrane spanning protein 193:230 a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand 193:340 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 1 34 theme serine 272:277 arg1 protease 279:286 a serine protease 270:286 a serine protease 270:286 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 8 35 theme full-length 1536:1546 arg1 form 1548:1551 the short, but not the full-length form 1513:1551 form 1548:1551 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 8 36 theme hRho 1466:1469 arg1 /RHBDF1 1471:1477 p100(hRho)/RHBDF1 1461:1477 p100(hRho)/RHBDF1 1461:1477 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 2 37 theme receptor 460:467 arg1 activation 469:478 consequent EGF receptor activation 445:478 consequent EGF receptor activation 445:478 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 4 38 theme long 705:708 arg1 protease 841:848 a serine protease 832:848 a serine protease 832:848 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 4 38 theme long 705:708 arg1 rhomboid 716:723 an atypically long human rhomboid 691:723 an atypically long human rhomboid 691:723 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 6 39 theme seven-transmembrane 1056:1074 arg1 rhomboid 1013:1020 This rhomboid 1008:1020 This rhomboid 1008:1020 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 6 39 theme seven-transmembrane 1056:1074 arg1 protein 1076:1082 a seven-transmembrane protein 1054:1082 a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface 1054:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 7 40 theme different 1294:1302 arg1 lengths 1304:1310 different lengths 1294:1310 different lengths 1294:1310 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 1 41 theme seven-transmembrane 195:213 arg1 protein 224:230 a seven-transmembrane spanning protein 193:230 a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand 193:340 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 1 41 theme seven-transmembrane 195:213 arg1 protease 279:286 a serine protease 270:286 a serine protease 270:286 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 1 41 theme seven-transmembrane 195:213 arg1 Rhomboid 183:190 Rhomboid 183:190 Rhomboid 183:190 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 2 42 theme EGF 456:458 arg1 activation 469:478 consequent EGF receptor activation 445:478 consequent EGF receptor activation 445:478 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 8 43 theme /RHBDF1 1471:1477 arg1 function 1449:1456 the function 1445:1456 the function of p100(hRho)/RHBDF1 in Drosophila 1445:1491 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 0 44 theme rhomboid 28:35 arg1 homolog 37:43 a human rhomboid homolog 20:43 a human rhomboid homolog 20:43 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 0 44 theme rhomboid 28:35 arg1 p100hRho/RHBDF1 46:60 p100hRho/RHBDF1 46:60 p100hRho/RHBDF1 46:60 Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands. 15965977 2 45 theme TGF-alpha-like 417:430 arg1 ligands 432:438 TGF-alpha-like ligands 417:438 TGF-alpha-like ligands 417:438 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 1 46 theme spanning 215:222 arg1 protein 224:230 a seven-transmembrane spanning protein 193:230 a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand 193:340 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 1 46 theme spanning 215:222 arg1 protease 279:286 a serine protease 270:286 a serine protease 270:286 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 1 46 theme spanning 215:222 arg1 Rhomboid 183:190 Rhomboid 183:190 Rhomboid 183:190 The activity of the TGF-alpha-like ligand Spitz in Drosophila depends on Rhomboid, a seven-transmembrane spanning protein that resides in the Golgi and acts as a serine protease to cleave Spitz, thereby releasing the soluble ligand. 15965977 2 47 theme consequent 445:454 arg1 activation 469:478 consequent EGF receptor activation 445:478 consequent EGF receptor activation 445:478 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 7 48 with forms 1283:1287 arg1 lengths 1304:1310 different lengths 1294:1310 different lengths 1294:1310 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 5 49 theme mouse 994:998 arg1 embryo 1000:1005 mouse embryo 994:1005 mouse embryo 994:1005 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 4 50 theme human 710:714 arg1 protease 841:848 a serine protease 832:848 a serine protease 832:848 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 4 50 theme human 710:714 arg1 rhomboid 716:723 an atypically long human rhomboid 691:723 an atypically long human rhomboid 691:723 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 6 51 theme cell 1246:1249 arg1 surface 1251:1257 the cell surface 1242:1257 the cell surface 1242:1257 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 5 52 with comparison 891:900 arg1 receptor 981:988 the EGF/TGF-alpha receptor 963:988 the EGF/TGF-alpha receptor 963:988 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 5 52 with comparison 891:900 arg1 HB-EGF 951:956 the TGF-alpha-related protein HB-EGF 921:956 the TGF-alpha-related protein HB-EGF 921:956 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 5 52 with comparison 891:900 arg1 TGF-alpha 907:915 TGF-alpha 907:915 TGF-alpha 907:915 We examined its tissue distribution, in comparison with TGF-alpha and the TGF-alpha-related protein HB-EGF, and the EGF/TGF-alpha receptor, in mouse embryo. 15965977 3 53 theme larger 485:490 arg1 number 492:497 The larger number 481:497 The larger number of TGF-alpha-like ligands in vertebrates 481:538 The larger number of TGF-alpha-like ligands in vertebrates raises the possibility that they too might be subject to regulation by rhomboid-like proteins. 15965977 6 54 theme sequence 1168:1175 arg1 half 1144:1147 half 1144:1147 half of the polypeptide sequence 1144:1175 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 6 55 theme polypeptide 1156:1166 arg1 sequence 1168:1175 the polypeptide sequence 1152:1175 the polypeptide sequence 1152:1175 This rhomboid, named p100(hRho) or RHBDF1, is a seven-transmembrane protein with a long N-terminal cytoplasmic extension that comprises half of the polypeptide sequence, and is found in the endoplasmic reticulum and Golgi, but not on the cell surface. 15965977 3 56 theme rhomboid-like 611:623 arg1 proteins 625:632 rhomboid-like proteins 611:632 rhomboid-like proteins 611:632 The larger number of TGF-alpha-like ligands in vertebrates raises the possibility that they too might be subject to regulation by rhomboid-like proteins. 15965977 8 57 contain has 1553:1555 arg1 form 1548:1551 the short, but not the full-length form 1513:1551 form 1548:1551 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 8 57 contain has 1553:1555 arg2 activity 1568:1575 functional activity 1557:1575 functional activity 1557:1575 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 8 57 contain has 1553:1555 arg1 short 1517:1521 short 1517:1521 short 1517:1521 Finally, we evaluated the function of p100(hRho)/RHBDF1 in Drosophila, demonstrating that the short, but not the full-length form has functional activity. 15965977 4 58 theme residues 766:773 arg1 absence 746:752 the absence 742:752 the absence of critical residues for serine protease activity 742:802 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 2 59 theme Several 343:349 arg1 rhomboids 351:359 Several rhomboids 343:359 Several rhomboids in Drosophila 343:373 Several rhomboids in Drosophila have been implicated in the processing of TGF-alpha-like ligands, and consequent EGF receptor activation. 15965977 4 60 theme protease 786:793 arg1 activity 795:802 serine protease activity 779:802 serine protease activity 779:802 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 3 61 theme TGF-alpha-like 502:515 arg1 ligands 517:523 TGF-alpha-like ligands 502:523 TGF-alpha-like ligands 502:523 The larger number of TGF-alpha-like ligands in vertebrates raises the possibility that they too might be subject to regulation by rhomboid-like proteins. 15965977 7 62 with interaction 1381:1391 arg1 ectodomain 1411:1420 the EGF core ectodomain 1398:1420 the EGF core ectodomain 1398:1420 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 9 63 theme rhomboid 1648:1655 arg1 family 1657:1662 the rhomboid family 1644:1662 the rhomboid family of regulatory proteins 1644:1685 The characterization of this protein extends our understanding of the rhomboid family of regulatory proteins. 15965977 7 64 theme EGF 1402:1404 arg1 ectodomain 1411:1420 the EGF core ectodomain 1398:1420 the EGF core ectodomain 1398:1420 It is expressed as two forms with different lengths, forms dimers and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. 15965977 4 65 theme serine 779:784 arg1 activity 795:802 serine protease activity 779:802 serine protease activity 779:802 We present the cDNA cloning and polypeptide sequence of an atypically long human rhomboid, which, based on the absence of critical residues for serine protease activity, is not predicted to act as a serine protease. 15965977 3 66 theme ligands 517:523 arg1 number 492:497 The larger number 481:497 The larger number of TGF-alpha-like ligands in vertebrates 481:538 The larger number of TGF-alpha-like ligands in vertebrates raises the possibility that they too might be subject to regulation by rhomboid-like proteins. 15965977 9 67 theme proteins 1678:1685 arg1 family 1657:1662 the rhomboid family 1644:1662 the rhomboid family of regulatory proteins 1644:1685 The characterization of this protein extends our understanding of the rhomboid family of regulatory proteins. 9689040 3 0 theme parent 487:492 arg1 body 494:497 the 201-residue remaining parent body 461:497 the 201-residue remaining parent body 461:497 Felicitously, during crystallization an amino-terminal segment was removed, apparently by a contaminating protease, allowing the 201-residue remaining parent body to crystallize. 9689040 7 1 theme beta- 873:877 arg1 structures 855:864 structures 855:864 structures of the beta- and gamma-chain carboxyl domains of human fibrinogen 855:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 5 2 theme P1 758:759 arg1 symmetry 748:755 noncrystallographic symmetry 728:755 noncrystallographic symmetry (P1 space group) 728:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 5 2 theme P1 758:759 arg1 group 767:771 P1 space group 758:771 P1 space group 758:771 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 0 3 from structure 8:16 arg1 fibrinogen-420 61:74 human fibrinogen-420 55:74 human fibrinogen-420 55:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 7 4 with Comparison 839:848 arg1 structures 855:864 structures 855:864 structures of the beta- and gamma-chain carboxyl domains of human fibrinogen 855:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 5 theme binding 950:956 arg1 neutral 979:985 neutral 979:985 neutral 979:985 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 5 theme binding 950:956 arg1 cleft 958:962 the binding cleft 946:962 the binding cleft 946:962 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 2 6 theme Pichia 313:318 arg1 pastoris 320:327 Pichia pastoris 313:327 Pichia pastoris cells 313:333 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 5 7 theme asymmetric 701:710 arg1 unit 712:715 the asymmetric unit 697:715 the asymmetric unit related by noncrystallographic symmetry (P1 space group) 697:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 2 8 theme alphaE 264:269 arg1 chain 271:275 the alphaE chain 260:275 the alphaE chain 260:275 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 2 9 theme pastoris 320:327 arg1 cells 329:333 Pichia pastoris cells 313:333 Pichia pastoris cells 313:333 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 5 10 theme related 717:723 arg1 unit 712:715 the asymmetric unit 697:715 the asymmetric unit related by noncrystallographic symmetry (P1 space group) 697:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 3 11 theme 201-residue 465:475 arg1 body 494:497 the 201-residue remaining parent body 461:497 the 201-residue remaining parent body 461:497 Felicitously, during crystallization an amino-terminal segment was removed, apparently by a contaminating protease, allowing the 201-residue remaining parent body to crystallize. 9689040 1 12 from fibrinogen-420 142:155 arg1 domain 124:129 a recombinant alphaEC domain 102:129 a recombinant alphaEC domain from human fibrinogen-420 102:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 1 12 from fibrinogen-420 142:155 arg1 structure 89:97 The crystal structure 77:97 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 77:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 7 13 dep beta- 873:877 arg1 the 869:871 the 869:871 the 869:871 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 3 14 theme amino-terminal 376:389 arg1 segment 391:397 an amino-terminal segment 373:397 an amino-terminal segment 373:397 Felicitously, during crystallization an amino-terminal segment was removed, apparently by a contaminating protease, allowing the 201-residue remaining parent body to crystallize. 9689040 5 15 theme space 761:765 arg1 symmetry 748:755 noncrystallographic symmetry 728:755 noncrystallographic symmetry (P1 space group) 728:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 5 15 theme space 761:765 arg1 group 767:771 P1 space group 758:771 P1 space group 758:771 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 7 16 theme fibrinogen 921:930 arg1 fibrinogen 921:930 human fibrinogen 915:930 human fibrinogen 915:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 16 theme fibrinogen 921:930 arg1 beta- 873:877 beta- 873:877 beta- 873:877 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 16 theme fibrinogen 921:930 arg1 domains 904:910 gamma-chain carboxyl domains 883:910 gamma-chain carboxyl domains 883:910 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 5 17 dep averaging 639:647 arg1 made 649:652 made 649:652 averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group) 639:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 7 18 theme human 915:919 arg1 fibrinogen 921:930 human fibrinogen 915:930 human fibrinogen 915:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 1 19 theme crystal 81:87 arg1 structure 89:97 The crystal structure 77:97 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 77:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 0 20 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420 0:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 7 21 theme other 1070:1074 arg1 domains 1076:1082 those other domains 1064:1082 those other domains 1064:1082 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 5 22 theme electron 579:586 arg1 result 629:634 a result 627:634 a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group) 627:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 5 22 theme electron 579:586 arg1 density 588:594 The electron density 575:594 The electron density 575:594 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 8 23 theme sialic 1186:1191 arg1 acid 1193:1196 sialic acid 1186:1196 sialic acid 1186:1196 Nonetheless, the cleft is clearly evident, and the possibility of binding a carbohydrate ligand like sialic acid has been considered. 9689040 8 24 theme carbohydrate 1161:1172 arg1 ligand 1174:1179 a carbohydrate ligand 1159:1179 a carbohydrate ligand like sialic acid 1159:1196 Nonetheless, the cleft is clearly evident, and the possibility of binding a carbohydrate ligand like sialic acid has been considered. 9689040 4 25 theme molecular 552:560 arg1 replacement 562:572 molecular replacement 552:572 molecular replacement 552:572 An x-ray structure was determined by molecular replacement. 9689040 2 26 theme chain 271:275 arg1 domain 250:255 the carboxyl domain 237:255 the carboxyl domain of the alphaE chain 237:275 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 2 26 theme chain 271:275 arg1 chain 271:275 the alphaE chain 260:275 the alphaE chain 260:275 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 0 27 theme alphaEC 35:41 arg1 domain 43:48 a recombinant alphaEC domain 21:48 a recombinant alphaEC domain from human fibrinogen-420 21:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 3 28 theme remaining 477:485 arg1 body 494:497 the 201-residue remaining parent body 461:497 the 201-residue remaining parent body 461:497 Felicitously, during crystallization an amino-terminal segment was removed, apparently by a contaminating protease, allowing the 201-residue remaining parent body to crystallize. 9689040 7 29 theme gamma-chain 883:893 arg1 fibrinogen 921:930 human fibrinogen 915:930 human fibrinogen 915:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 29 theme gamma-chain 883:893 arg1 domains 904:910 gamma-chain carboxyl domains 883:910 gamma-chain carboxyl domains 883:910 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 1 30 theme A 200:200 arg1 resolution 182:191 a resolution 180:191 a resolution of 2.1 A 180:200 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 0 31 theme recombinant 23:33 arg1 domain 43:48 a recombinant alphaEC domain 21:48 a recombinant alphaEC domain from human fibrinogen-420 21:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 7 32 theme sort 1050:1053 arg1 peptides 1034:1041 Gly-Pro-Arg or Gly-His-Arg peptides 1007:1041 Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains 1007:1082 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 0 33 from fibrinogen-420 61:74 arg1 domain 43:48 a recombinant alphaEC domain 21:48 a recombinant alphaEC domain from human fibrinogen-420 21:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 0 33 from fibrinogen-420 61:74 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420 0:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 3 34 theme contaminating 428:440 arg1 protease 442:449 a contaminating protease 426:449 a contaminating protease 426:449 Felicitously, during crystallization an amino-terminal segment was removed, apparently by a contaminating protease, allowing the 201-residue remaining parent body to crystallize. 9689040 7 35 theme Gly-Pro-Arg 1007:1017 arg1 peptides 1034:1041 Gly-Pro-Arg or Gly-His-Arg peptides 1007:1041 Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains 1007:1082 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 1 36 theme recombinant 104:114 arg1 domain 124:129 a recombinant alphaEC domain 102:129 a recombinant alphaEC domain from human fibrinogen-420 102:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 0 37 theme domain 43:48 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420 0:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 1 38 theme alphaEC 116:122 arg1 domain 124:129 a recombinant alphaEC domain 102:129 a recombinant alphaEC domain from human fibrinogen-420 102:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 7 39 theme carboxyl 895:902 arg1 fibrinogen 921:930 human fibrinogen 915:930 human fibrinogen 915:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 39 theme carboxyl 895:902 arg1 domains 904:910 gamma-chain carboxyl domains 883:910 gamma-chain carboxyl domains 883:910 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 4 40 theme x-ray 518:522 arg1 structure 524:532 An x-ray structure 515:532 An x-ray structure 515:532 An x-ray structure was determined by molecular replacement. 9689040 2 41 theme carboxyl 241:248 arg1 domain 250:255 the carboxyl domain 237:255 the carboxyl domain of the alphaE chain 237:275 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 2 41 theme carboxyl 241:248 arg1 chain 271:275 the alphaE chain 260:275 the alphaE chain 260:275 The protein, which corresponds to the carboxyl domain of the alphaE chain, was expressed in and purified from Pichia pastoris cells. 9689040 1 42 theme domain 124:129 arg1 structure 89:97 The crystal structure 77:97 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 77:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 7 43 theme domains 904:910 arg1 structures 855:864 structures 855:864 structures of the beta- and gamma-chain carboxyl domains of human fibrinogen 855:930 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 7 44 theme Gly-His-Arg 1022:1032 arg1 peptides 1034:1041 Gly-Pro-Arg or Gly-His-Arg peptides 1007:1041 Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains 1007:1082 Comparison with structures of the beta- and gamma-chain carboxyl domains of human fibrinogen revealed that the binding cleft is essentially neutral and should not bind Gly-Pro-Arg or Gly-His-Arg peptides of the sort bound by those other domains. 9689040 6 45 theme sugar 813:817 arg1 cluster 819:825 an asparagine-linked sugar cluster 792:825 an asparagine-linked sugar cluster 792:825 Virtually all of an asparagine-linked sugar cluster is present. 9689040 5 46 theme noncrystallographic 728:746 arg1 symmetry 748:755 noncrystallographic symmetry 728:755 noncrystallographic symmetry (P1 space group) 728:772 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 5 46 theme noncrystallographic 728:746 arg1 group 767:771 P1 space group 758:771 P1 space group 758:771 The electron density was clearly defined, partly as a result of averaging made possible by there being eight molecules in the asymmetric unit related by noncrystallographic symmetry (P1 space group). 9689040 6 47 theme asparagine-linked 795:811 arg1 cluster 819:825 an asparagine-linked sugar cluster 792:825 an asparagine-linked sugar cluster 792:825 Virtually all of an asparagine-linked sugar cluster is present. 9689040 1 48 theme human 136:140 arg1 fibrinogen-420 142:155 human fibrinogen-420 136:155 human fibrinogen-420 136:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 1 49 from structure 89:97 arg1 fibrinogen-420 142:155 human fibrinogen-420 136:155 human fibrinogen-420 136:155 The crystal structure of a recombinant alphaEC domain from human fibrinogen-420 has been determined at a resolution of 2.1 A. 9689040 0 50 theme human 55:59 arg1 fibrinogen-420 61:74 human fibrinogen-420 55:74 human fibrinogen-420 55:74 Crystal structure of a recombinant alphaEC domain from human fibrinogen-420. 9689040 6 51 link asparagine-linked 795:811 arg1 cluster 819:825 an asparagine-linked sugar cluster 792:825 an asparagine-linked sugar cluster 792:825 Virtually all of an asparagine-linked sugar cluster is present. 27238017 6 0 theme biochemical 998:1008 arg1 characterizations 1010:1026 Our structural and biochemical characterizations 979:1026 Our structural and biochemical characterizations 979:1026 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 3 1 theme NPC1 466:469 arg1 structure 391:399 a 4.4 Å structure 383:399 a 4.4 Å structure of full-length human NPC1 383:425 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 1 theme NPC1 466:469 arg1 reconstruction 448:461 a low-resolution reconstruction 431:461 a low-resolution reconstruction of NPC1 431:469 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 1 2 theme key 208:210 arg1 products 195:202 gene products 190:202 gene products 190:202 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 2 theme key 208:210 arg1 players 212:218 key players 208:218 key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol 208:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 0 3 theme Cholesterol 61:71 arg1 Transfer 73:80 the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer 25:80 the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer 25:80 Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection. 27238017 4 4 theme distinct 644:651 arg1 A 669:669 A 669:669 A (also designated NTD) 669:691 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 4 theme distinct 644:651 arg1 C 694:694 C 694:694 C 694:694 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 4 theme distinct 644:651 arg1 domains 661:667 three distinct lumenal domains 638:667 three distinct lumenal domains A (also designated NTD), C, and I. 638:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 4 theme distinct 644:651 arg1 I. 701:702 I. 701:702 I. 701:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 0 5 theme Ebola 86:90 arg1 Infection 92:100 Ebola Infection 86:100 Ebola Infection 86:100 Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection. 27238017 3 6 with complex 474:480 arg1 glycoprotein 499:510 the cleaved glycoprotein 487:510 the cleaved glycoprotein (GPcl) of EBOV 487:525 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 6 with complex 474:480 arg1 GPcl 513:516 GPcl 513:516 GPcl 513:516 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 5 7 theme trimeric 912:919 arg1 EBOV-GPcl 921:929 A trimeric EBOV-GPcl 910:929 A trimeric EBOV-GPcl 910:929 A trimeric EBOV-GPcl binds to one NPC1 monomer through the domain C. 27238017 4 8 theme lumenal 653:659 arg1 A 669:669 A 669:669 A (also designated NTD) 669:691 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 8 theme lumenal 653:659 arg1 C 694:694 C 694:694 C 694:694 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 8 theme lumenal 653:659 arg1 domains 661:667 three distinct lumenal domains 638:667 three distinct lumenal domains A (also designated NTD), C, and I. 638:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 8 theme lumenal 653:659 arg1 I. 701:702 I. 701:702 I. 701:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 3 9 theme low-resolution 433:446 arg1 reconstruction 448:461 a low-resolution reconstruction 431:461 a low-resolution reconstruction of NPC1 431:469 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 1 10 theme Niemann-Pick 103:114 arg1 NPC 132:134 NPC 132:134 NPC 132:134 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 10 theme Niemann-Pick 103:114 arg1 C 129:129 Niemann-Pick disease type C 103:129 Niemann-Pick disease type C (NPC) 103:135 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 6 11 theme mechanistic 1063:1073 arg1 understanding 1075:1087 mechanistic understanding 1063:1087 mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection 1063:1168 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 6 12 theme cholesterol 1120:1130 arg1 trafficking 1132:1142 NPC1-mediated intracellular cholesterol trafficking 1092:1142 NPC1-mediated intracellular cholesterol trafficking 1092:1142 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 6 13 theme intracellular 1106:1118 arg1 trafficking 1132:1142 NPC1-mediated intracellular cholesterol trafficking 1092:1142 NPC1-mediated intracellular cholesterol trafficking 1092:1142 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 3 14 theme cleaved 491:497 arg1 glycoprotein 499:510 the cleaved glycoprotein 487:510 the cleaved glycoprotein (GPcl) of EBOV 487:525 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 14 theme cleaved 491:497 arg1 GPcl 513:516 GPcl 513:516 GPcl 513:516 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 6 15 theme important 1039:1047 arg1 framework 1049:1057 an important framework 1036:1057 an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection 1036:1168 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 6 16 theme infection 1160:1168 arg1 understanding 1075:1087 mechanistic understanding 1063:1087 mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection 1063:1168 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 0 17 theme Structural 0:9 arg1 Insights 11:18 Structural Insights 0:18 Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection. 0:101 Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection. 27238017 1 18 theme endosomal/lysosomal 227:245 arg1 egress 247:252 the endosomal/lysosomal egress 223:252 the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol 223:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 3 19 dep present 375:381 arg1 both 528:531 both 528:531 both 528:531 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 20 theme EBOV 522:525 arg1 glycoprotein 499:510 the cleaved glycoprotein 487:510 the cleaved glycoprotein (GPcl) of EBOV 487:525 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 20 theme EBOV 522:525 arg1 GPcl 513:516 GPcl 513:516 GPcl 513:516 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 21 theme 4.4 Å 385:389 arg1 structure 391:399 a 4.4 Å structure 383:399 a 4.4 Å structure of full-length human NPC1 383:425 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 6 22 theme Ebola 1148:1152 arg1 infection 1160:1168 Ebola virus infection 1148:1168 Ebola virus infection 1148:1168 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 3 23 theme NPC1 422:425 arg1 structure 391:399 a 4.4 Å structure 383:399 a 4.4 Å structure of full-length human NPC1 383:425 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 23 theme NPC1 422:425 arg1 reconstruction 448:461 a low-resolution reconstruction 431:461 a low-resolution reconstruction of NPC1 431:469 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 4 24 theme designated 677:686 arg1 A 669:669 A 669:669 A (also designated NTD) 669:691 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 24 theme designated 677:686 arg1 NTD 688:690 also designated NTD 672:690 also designated NTD 672:690 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 3 25 attach present 375:381 arg2 we 372:373 we 372:373 we 372:373 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 25 attach present 375:381 arg1 complex 474:480 complex 474:480 complex with the cleaved glycoprotein (GPcl) of EBOV 474:525 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 4 26 dep domains 661:667 arg1 domains 661:667 three distinct lumenal domains 638:667 three distinct lumenal domains A (also designated NTD), C, and I. 638:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 26 dep domains 661:667 arg1 A 669:669 A 669:669 A (also designated NTD) 669:691 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 26 dep domains 661:667 arg1 NTD 688:690 also designated NTD 672:690 also designated NTD 672:690 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 26 dep domains 661:667 arg1 C 694:694 C 694:694 C 694:694 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 26 dep domains 661:667 arg1 I. 701:702 I. 701:702 I. 701:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 6 27 theme virus 1154:1158 arg1 infection 1160:1168 Ebola virus infection 1148:1168 Ebola virus infection 1148:1168 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 1 28 theme low-density 257:267 arg1 cholesterol 289:299 low-density lipoprotein-derived cholesterol 257:299 low-density lipoprotein-derived cholesterol 257:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 4 29 theme transmembrane 605:617 arg1 TMs 629:631 TMs 629:631 TMs 629:631 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 29 theme transmembrane 605:617 arg1 segments 619:626 13 transmembrane segments 602:626 13 transmembrane segments (TMs) 602:632 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 30 theme resistance-nodulation-cell 731:756 arg1 fold 767:770 a typical resistance-nodulation-cell division fold 721:770 a typical resistance-nodulation-cell division fold 721:770 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 31 dep TMs 704:706 arg1 2-13 708:711 2-13 708:711 2-13 708:711 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 3 32 theme full-length 404:414 arg1 NPC1 422:425 full-length human NPC1 404:425 full-length human NPC1 404:425 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 1 33 theme lipoprotein-derived 269:287 arg1 cholesterol 289:299 low-density lipoprotein-derived cholesterol 257:299 low-density lipoprotein-derived cholesterol 257:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 34 from players 212:218 arg1 egress 247:252 the endosomal/lysosomal egress 223:252 the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol 223:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 4 35 theme typical 723:729 arg1 fold 767:770 a typical resistance-nodulation-cell division fold 721:770 a typical resistance-nodulation-cell division fold 721:770 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 3 36 theme human 416:420 arg1 NPC1 422:425 full-length human NPC1 404:425 full-length human NPC1 404:425 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 1 37 theme cholesterol 289:299 arg1 egress 247:252 the endosomal/lysosomal egress 223:252 the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol 223:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 4 38 theme sterol-sensing 808:821 arg1 domain 823:828 the sterol-sensing domain 804:828 the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling 804:907 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg1 NPC1 588:591 NPC1 588:591 NPC1 588:591 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg2 segments 619:626 13 transmembrane segments 602:626 13 transmembrane segments (TMs) 602:632 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg2 A 669:669 A 669:669 A (also designated NTD) 669:691 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg2 TMs 629:631 TMs 629:631 TMs 629:631 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg2 domains 661:667 three distinct lumenal domains 638:667 three distinct lumenal domains A (also designated NTD), C, and I. 638:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg2 C 694:694 C 694:694 C 694:694 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 4 39 contain contains 593:600 arg2 I. 701:702 I. 701:702 I. 701:702 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 2 40 theme intracellular 319:331 arg1 receptor 333:340 the intracellular receptor 315:340 the intracellular receptor for Ebola virus (EBOV) 315:363 NPC1 is also the intracellular receptor for Ebola virus (EBOV). 27238017 2 40 theme intracellular 319:331 arg1 NPC1 302:305 NPC1 302:305 NPC1 302:305 NPC1 is also the intracellular receptor for Ebola virus (EBOV). 27238017 4 41 theme division 758:765 arg1 fold 767:770 a typical resistance-nodulation-cell division fold 721:770 a typical resistance-nodulation-cell division fold 721:770 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 1 42 link lipoprotein-derived 269:287 arg1 cholesterol 289:299 low-density lipoprotein-derived cholesterol 257:299 low-density lipoprotein-derived cholesterol 257:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 43 theme disease 116:122 arg1 NPC 132:134 NPC 132:134 NPC 132:134 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 43 theme disease 116:122 arg1 C 129:129 Niemann-Pick disease type C 103:129 Niemann-Pick disease type C (NPC) 103:135 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 4 44 theme several 843:849 arg1 proteins 851:858 several proteins 843:858 several proteins involved in cholesterol metabolism and signaling 843:907 NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. 27238017 1 45 theme type 124:127 arg1 NPC 132:134 NPC 132:134 NPC 132:134 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 45 theme type 124:127 arg1 C 129:129 Niemann-Pick disease type C 103:129 Niemann-Pick disease type C (NPC) 103:135 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 6 46 theme NPC1-mediated 1092:1104 arg1 trafficking 1132:1142 NPC1-mediated intracellular cholesterol trafficking 1092:1142 NPC1-mediated intracellular cholesterol trafficking 1092:1142 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 5 47 theme NPC1 944:947 arg1 monomer 949:955 one NPC1 monomer 940:955 one NPC1 monomer 940:955 A trimeric EBOV-GPcl binds to one NPC1 monomer through the domain C. 27238017 1 48 from mutations 156:164 arg1 NPC2 178:181 NPC2 178:181 NPC2 178:181 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 48 from mutations 156:164 arg1 NPC1 169:172 NPC1 169:172 NPC1 169:172 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 2 49 theme Ebola 346:350 arg1 EBOV 359:362 EBOV 359:362 EBOV 359:362 NPC1 is also the intracellular receptor for Ebola virus (EBOV). 27238017 2 49 theme Ebola 346:350 arg1 virus 352:356 Ebola virus 346:356 Ebola virus (EBOV) 346:363 NPC1 is also the intracellular receptor for Ebola virus (EBOV). 27238017 6 50 theme trafficking 1132:1142 arg1 understanding 1075:1087 mechanistic understanding 1063:1087 mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection 1063:1168 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 1 51 theme gene 190:193 arg1 products 195:202 gene products 190:202 gene products 190:202 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 1 51 theme gene 190:193 arg1 players 212:218 key players 208:218 key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol 208:299 Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. 27238017 0 52 theme -Mediated 51:59 arg1 Transfer 73:80 the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer 25:80 the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer 25:80 Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection. 27238017 6 53 theme structural 983:992 arg1 characterizations 1010:1026 Our structural and biochemical characterizations 979:1026 Our structural and biochemical characterizations 979:1026 Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection. 27238017 3 54 theme single-particle 547:561 arg1 cryomicroscopy 572:585 single-particle electron cryomicroscopy 547:585 single-particle electron cryomicroscopy 547:585 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 55 theme electron 563:570 arg1 cryomicroscopy 572:585 single-particle electron cryomicroscopy 547:585 single-particle electron cryomicroscopy 547:585 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 56 gly glycoprotein 499:510 arg1 glycoprotein 499:510 the cleaved glycoprotein 487:510 the cleaved glycoprotein (GPcl) of EBOV 487:525 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 27238017 3 56 gly glycoprotein 499:510 arg1 GPcl 513:516 GPcl 513:516 GPcl 513:516 Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. 22128166 6 0 from defects 870:876 arg1 diseases 900:907 the above diseases 890:907 the above diseases 890:907 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 1 1 theme Morquio 132:138 arg1 diseases 184:191 autosomal recessive lysosomal storage diseases 146:191 autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism 146:248 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 1 theme Morquio 132:138 arg1 gangliosidosis 113:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 1 theme Morquio 132:138 arg1 B 140:140 Morquio B 132:140 Morquio B 132:140 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 0 2 theme gangliosidosis 68:81 arg1 basis 55:59 structural basis 44:59 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 0:105 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 2 3 theme enzyme 351:356 arg1 β-d-galactosidase 358:374 the lysosomal enzyme β-d-galactosidase 337:374 the lysosomal enzyme β-d-galactosidase (β-Gal) 337:382 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 2 3 theme enzyme 351:356 arg1 β-Gal 377:381 β-Gal 377:381 β-Gal 377:381 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 0 4 theme B 95:95 arg1 diseases 97:104 morquio B diseases 87:104 morquio B diseases 87:104 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 6 5 theme β-Gal 881:885 arg1 defects 870:876 the molecular defects 856:876 the molecular defects of β-Gal in the above diseases 856:907 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 2 6 from deficiencies 321:332 arg1 β-Gal 377:381 β-Gal 377:381 β-Gal 377:381 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 2 6 from deficiencies 321:332 arg1 β-d-galactosidase 358:374 the lysosomal enzyme β-d-galactosidase 337:374 the lysosomal enzyme β-d-galactosidase (β-Gal) 337:382 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 2 6 from deficiencies 321:332 arg1 sulfate 469:475 keratan sulfate 461:475 keratan sulfate 461:475 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 2 6 from deficiencies 321:332 arg1 ganglioside 444:454 G(M1) ganglioside 438:454 G(M1) ganglioside 438:454 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 1 7 theme skeletal 254:261 arg1 abnormalities 263:275 skeletal abnormalities 254:275 skeletal abnormalities 254:275 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 0 8 theme morquio 87:93 arg1 diseases 97:104 morquio B diseases 87:104 morquio B diseases 87:104 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 5 9 theme catalytic 757:765 arg1 domain 778:783 a catalytic TIM barrel domain 755:783 a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2 755:821 Human β-Gal is composed of a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2. 22128166 2 10 theme lysosomal 341:349 arg1 β-d-galactosidase 358:374 the lysosomal enzyme β-d-galactosidase 337:374 the lysosomal enzyme β-d-galactosidase (β-Gal) 337:382 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 2 10 theme lysosomal 341:349 arg1 β-Gal 377:381 β-Gal 377:381 β-Gal 377:381 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 0 11 theme diseases 97:104 arg1 basis 55:59 structural basis 44:59 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 0:105 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 6 12 theme molecular 860:868 arg1 defects 870:876 the molecular defects 856:876 the molecular defects of β-Gal in the above diseases 856:907 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 4 13 from complex 634:640 arg1 structures 605:614 the crystal structures 593:614 the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin 593:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 2 14 theme G 438:438 arg1 ganglioside 444:454 G(M1) ganglioside 438:454 G(M1) ganglioside 438:454 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 3 15 link β-linked 547:554 arg1 residues 566:573 terminal β-linked galactose residues 538:573 terminal β-linked galactose residues 538:573 β-Gal is an exoglycosidase that catalyzes the hydrolysis of terminal β-linked galactose residues. 22128166 3 16 theme terminal 538:545 arg1 residues 566:573 terminal β-linked galactose residues 538:573 terminal β-linked galactose residues 538:573 β-Gal is an exoglycosidase that catalyzes the hydrolysis of terminal β-linked galactose residues. 22128166 1 17 theme recessive 156:164 arg1 diseases 184:191 autosomal recessive lysosomal storage diseases 146:191 autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism 146:248 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 17 theme recessive 156:164 arg1 gangliosidosis 113:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 17 theme recessive 156:164 arg1 B 140:140 Morquio B 132:140 Morquio B 132:140 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 0 18 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 0:105 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 3 19 theme β-linked 547:554 arg1 residues 566:573 terminal β-linked galactose residues 538:573 terminal β-linked galactose residues 538:573 β-Gal is an exoglycosidase that catalyzes the hydrolysis of terminal β-linked galactose residues. 22128166 4 20 from β-Gal 625:629 arg1 complex 634:640 complex 634:640 complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin 634:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 1 21 theme lysosomal 166:174 arg1 diseases 184:191 autosomal recessive lysosomal storage diseases 146:191 autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism 146:248 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 21 theme lysosomal 166:174 arg1 gangliosidosis 113:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 21 theme lysosomal 166:174 arg1 B 140:140 Morquio B 132:140 Morquio B 132:140 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 2 22 theme substrates 426:435 arg1 accumulations 399:411 accumulations 399:411 accumulations of the β-Gal substrates 399:435 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 1 23 theme storage 176:182 arg1 diseases 184:191 autosomal recessive lysosomal storage diseases 146:191 autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism 146:248 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 23 theme storage 176:182 arg1 gangliosidosis 113:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 23 theme storage 176:182 arg1 B 140:140 Morquio B 132:140 Morquio B 132:140 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 2 24 theme β-Gal 420:424 arg1 substrates 426:435 the β-Gal substrates 416:435 the β-Gal substrates 416:435 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 5 25 theme TIM 767:769 arg1 domain 778:783 a catalytic TIM barrel domain 755:783 a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2 755:821 Human β-Gal is composed of a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2. 22128166 4 26 from structures 605:614 arg1 complex 634:640 complex 634:640 complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin 634:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 4 27 theme β-Gal 625:629 arg1 structures 605:614 the crystal structures 593:614 the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin 593:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 0 28 theme β-galactosidase 27:41 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 0:105 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 6 29 theme disease-causing 914:928 arg1 mutations 930:938 the disease-causing mutations 910:938 the disease-causing mutations 910:938 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 5 30 theme barrel 771:776 arg1 domain 778:783 a catalytic TIM barrel domain 755:783 a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2 755:821 Human β-Gal is composed of a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2. 22128166 4 31 with complex 634:640 arg1 galactose 669:677 its catalytic product galactose 647:677 its catalytic product galactose 647:677 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 4 31 with complex 634:640 arg1 1-deoxygalactonojirimycin 701:725 its inhibitor 1-deoxygalactonojirimycin 687:725 its inhibitor 1-deoxygalactonojirimycin 687:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 0 32 theme human 21:25 arg1 β-galactosidase 27:41 human β-galactosidase 21:41 human β-galactosidase 21:41 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 4 33 theme human 619:623 arg1 β-Gal 625:629 human β-Gal 619:629 human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin 619:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 2 34 theme M1 440:441 arg1 ganglioside 444:454 G(M1) ganglioside 438:454 G(M1) ganglioside 438:454 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 3 35 theme galactose 556:564 arg1 residues 566:573 terminal β-linked galactose residues 538:573 terminal β-linked galactose residues 538:573 β-Gal is an exoglycosidase that catalyzes the hydrolysis of terminal β-linked galactose residues. 22128166 7 36 theme possible 1003:1010 arg1 causes 1012:1017 the possible causes 999:1017 the possible causes of the diseases 999:1033 Finally, the possible causes of the diseases are discussed. 22128166 1 37 theme G 107:107 arg1 diseases 184:191 autosomal recessive lysosomal storage diseases 146:191 autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism 146:248 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 37 theme G 107:107 arg1 gangliosidosis 113:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis 107:126 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 1 37 theme G 107:107 arg1 B 140:140 Morquio B 132:140 Morquio B 132:140 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 0 38 theme structural 44:53 arg1 basis 55:59 structural basis 44:59 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 0:105 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 7 39 theme diseases 1026:1033 arg1 causes 1012:1017 the possible causes 999:1017 the possible causes of the diseases 999:1033 Finally, the possible causes of the diseases are discussed. 22128166 3 40 theme residues 566:573 arg1 hydrolysis 524:533 the hydrolysis 520:533 the hydrolysis of terminal β-linked galactose residues 520:573 β-Gal is an exoglycosidase that catalyzes the hydrolysis of terminal β-linked galactose residues. 22128166 6 41 theme structural 832:841 arg1 insight 843:849 structural insight 832:849 structural insight into the molecular defects of β-Gal in the above diseases 832:907 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 6 42 theme three-dimensional 961:977 arg1 structure 979:987 the three-dimensional structure 957:987 the three-dimensional structure 957:987 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 4 43 theme inhibitor 691:699 arg1 1-deoxygalactonojirimycin 701:725 its inhibitor 1-deoxygalactonojirimycin 687:725 its inhibitor 1-deoxygalactonojirimycin 687:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 1 44 theme neurodegenerative 211:227 arg1 disorder 229:236 a neurodegenerative disorder 209:236 a neurodegenerative disorder 209:236 G(M1) gangliosidosis and Morquio B are autosomal recessive lysosomal storage diseases associated with a neurodegenerative disorder or dwarfism and skeletal abnormalities, respectively. 22128166 0 45 theme Gm1 64:66 arg1 gangliosidosis 68:81 Gm1 gangliosidosis 64:81 Gm1 gangliosidosis 64:81 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 0 46 dep structure 8:16 arg1 basis 55:59 structural basis 44:59 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 0:105 Crystal structure of human β-galactosidase: structural basis of Gm1 gangliosidosis and morquio B diseases. 22128166 4 47 theme product 661:667 arg1 galactose 669:677 its catalytic product galactose 647:677 its catalytic product galactose 647:677 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 4 48 theme crystal 597:603 arg1 structures 605:614 the crystal structures 593:614 the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin 593:725 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 2 49 theme keratan 461:467 arg1 sulfate 469:475 keratan sulfate 461:475 keratan sulfate 461:475 These diseases are caused by deficiencies in the lysosomal enzyme β-d-galactosidase (β-Gal), which lead to accumulations of the β-Gal substrates, G(M1) ganglioside, and keratan sulfate. 22128166 6 50 theme above 894:898 arg1 diseases 900:907 the above diseases 890:907 the above diseases 890:907 To gain structural insight into the molecular defects of β-Gal in the above diseases, the disease-causing mutations were mapped onto the three-dimensional structure. 22128166 4 51 theme catalytic 651:659 arg1 galactose 669:677 its catalytic product galactose 647:677 its catalytic product galactose 647:677 This study shows the crystal structures of human β-Gal in complex with its catalytic product galactose or with its inhibitor 1-deoxygalactonojirimycin. 22128166 5 52 theme Human 728:732 arg1 β-Gal 734:738 Human β-Gal 728:738 Human β-Gal 728:738 Human β-Gal is composed of a catalytic TIM barrel domain followed by β-domain 1 and β-domain 2. 24737316 2 0 theme endoplasmic 507:517 arg1 degradation 540:550 endoplasmic reticulum-associated degradation 507:550 endoplasmic reticulum-associated degradation 507:550 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 3 1 theme Japanese 745:752 arg1 patients 754:761 GM1 gangliosidosis Japanese patients 726:761 GM1 gangliosidosis Japanese patients 726:761 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 1 2 theme subsequent 249:258 arg1 degradation 293:303 subsequent endoplasmic reticulum-associated degradation 249:303 subsequent endoplasmic reticulum-associated degradation 249:303 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 3 3 from mutations 713:721 arg1 patients 754:761 GM1 gangliosidosis Japanese patients 726:761 GM1 gangliosidosis Japanese patients 726:761 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 4 4 theme β-Gal 864:868 arg1 inhibitors 850:859 two competitive inhibitors 834:859 two competitive inhibitors of β-Gal 834:868 We have also evaluated the PC effect of two competitive inhibitors of β-Gal. 24737316 1 5 from defect 163:168 arg1 β-galactosidase 187:201 the lysosomal β-galactosidase 173:201 the lysosomal β-galactosidase (β-Gal) 173:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 5 from defect 163:168 arg1 β-Gal 204:208 β-Gal 204:208 β-Gal 204:208 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 2 6 theme therapeutic 376:386 arg1 approach 388:395 a newly developed molecular therapeutic approach 348:395 a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 348:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 6 theme therapeutic 376:386 arg1 therapy 337:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 8 7 theme developed 1470:1478 arg1 PC 1480:1481 newly developed PC 1464:1481 newly developed PC 1464:1481 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 1 8 theme endoplasmic 260:270 arg1 degradation 293:303 subsequent endoplasmic reticulum-associated degradation 249:303 subsequent endoplasmic reticulum-associated degradation 249:303 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 3 9 theme GM1 726:728 arg1 patients 754:761 GM1 gangliosidosis Japanese patients 726:761 GM1 gangliosidosis Japanese patients 726:761 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 8 10 theme chaperoning 1449:1459 arg1 compounds 1483:1491 β-Gal selective chaperoning by newly developed PC compounds 1433:1491 β-Gal selective chaperoning by newly developed PC compounds 1433:1491 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 5 11 theme compounds 953:961 arg1 mechanism 934:942 the recognition mechanism 918:942 the recognition mechanism of these compounds 918:961 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 7 12 theme substituent 1354:1364 arg1 nature 1318:1323 nature 1318:1323 nature 1318:1323 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 12 theme substituent 1354:1364 arg1 length 1330:1335 length 1330:1335 length 1330:1335 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 12 theme substituent 1354:1364 arg1 structure 1268:1276 the mono- or bicyclic structure 1246:1276 the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent 1246:1364 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 12 theme substituent 1354:1364 arg1 orientation 1305:1315 orientation 1305:1315 orientation 1305:1315 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 1 13 theme reticulum-associated 272:291 arg1 degradation 293:303 subsequent endoplasmic reticulum-associated degradation 249:303 subsequent endoplasmic reticulum-associated degradation 249:303 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 5 14 theme recognition 922:932 arg1 mechanism 934:942 the recognition mechanism 918:942 the recognition mechanism of these compounds 918:961 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 1 15 theme GM1 75:77 arg1 gangliosidosis 79:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 15 theme GM1 75:77 arg1 diseases 140:147 autosomal recessive diseases 120:147 autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal) 120:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 15 theme GM1 75:77 arg1 disease 108:114 Morquio B disease 98:114 Morquio B disease 98:114 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 2 16 theme chaperone 322:330 arg1 approach 388:395 a newly developed molecular therapeutic approach 348:395 a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 348:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 16 theme chaperone 322:330 arg1 therapy 337:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 17 theme molecular 366:374 arg1 approach 388:395 a newly developed molecular therapeutic approach 348:395 a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 348:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 17 theme molecular 366:374 arg1 therapy 337:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 4 18 theme competitive 838:848 arg1 inhibitors 850:859 two competitive inhibitors 834:859 two competitive inhibitors of β-Gal 834:868 We have also evaluated the PC effect of two competitive inhibitors of β-Gal. 24737316 6 19 theme active 1044:1049 arg1 β-Gal 1059:1063 β-Gal 1059:1063 β-Gal 1059:1063 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 6 19 theme active 1044:1049 arg1 site 1051:1054 the active site 1040:1054 the active site of β-Gal 1040:1063 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 2 20 theme Pharmacological 306:320 arg1 approach 388:395 a newly developed molecular therapeutic approach 348:395 a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 348:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 20 theme Pharmacological 306:320 arg1 therapy 337:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 6 21 theme β-Gal 1059:1063 arg1 β-Gal 1059:1063 β-Gal 1059:1063 β-Gal 1059:1063 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 6 21 theme β-Gal 1059:1063 arg1 site 1051:1054 the active site 1040:1054 the active site of β-Gal 1040:1063 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 7 22 theme enzyme 1180:1185 arg1 selectivity 1187:1197 the enzyme selectivity 1176:1197 the enzyme selectivity 1176:1197 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 5 23 theme detailed 894:901 arg1 view 910:913 a detailed atomic view 892:913 a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues 892:1015 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 7 24 theme core 1285:1288 arg1 nature 1318:1323 nature 1318:1323 nature 1318:1323 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 24 theme core 1285:1288 arg1 length 1330:1335 length 1330:1335 length 1330:1335 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 24 theme core 1285:1288 arg1 structure 1268:1276 the mono- or bicyclic structure 1246:1276 the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent 1246:1364 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 24 theme core 1285:1288 arg1 orientation 1305:1315 orientation 1305:1315 orientation 1305:1315 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 6 25 theme sugar-mimicking 1074:1088 arg1 moiety 1090:1095 the sugar-mimicking moiety 1070:1095 the sugar-mimicking moiety making hydrogen bonds to active site residues 1070:1141 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 1 26 theme lysosomal 177:185 arg1 β-galactosidase 187:201 the lysosomal β-galactosidase 173:201 the lysosomal β-galactosidase (β-Gal) 173:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 26 theme lysosomal 177:185 arg1 β-Gal 204:208 β-Gal 204:208 β-Gal 204:208 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 3 27 from patients 754:761 arg1 properties 639:648 the enzymological properties 621:648 the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T) 621:791 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 8 28 from understanding 1389:1401 arg1 mechanism 1410:1418 the mechanism 1406:1418 the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds 1406:1491 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 1 29 theme Morquio 98:104 arg1 diseases 140:147 autosomal recessive diseases 120:147 autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal) 120:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 29 theme Morquio 98:104 arg1 gangliosidosis 79:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 29 theme Morquio 98:104 arg1 disease 108:114 Morquio B disease 98:114 Morquio B disease 98:114 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 8 30 theme compounds 1483:1491 arg1 action 1423:1428 action 1423:1428 action of β-Gal selective chaperoning by newly developed PC compounds 1423:1491 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 2 31 theme molecule 412:419 arg1 ligands 421:427 small molecule ligands 406:427 small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 406:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 0 32 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of pharmacological chaperoning for human β-galactosidase. 0:73 Structural basis of pharmacological chaperoning for human β-galactosidase. 24737316 4 33 theme inhibitors 850:859 arg1 effect 824:829 the PC effect 817:829 the PC effect of two competitive inhibitors of β-Gal 817:868 We have also evaluated the PC effect of two competitive inhibitors of β-Gal. 24737316 1 34 theme B 106:106 arg1 diseases 140:147 autosomal recessive diseases 120:147 autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal) 120:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 34 theme B 106:106 arg1 gangliosidosis 79:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 34 theme B 106:106 arg1 disease 108:114 Morquio B disease 98:114 Morquio B disease 98:114 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 2 35 theme small 406:410 arg1 ligands 421:427 small molecule ligands 406:427 small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 406:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 0 36 theme pharmacological 20:34 arg1 basis 11:15 Structural basis 0:15 Structural basis of pharmacological chaperoning for human β-galactosidase. 0:73 Structural basis of pharmacological chaperoning for human β-galactosidase. 24737316 5 37 from view 910:913 arg1 comparison 966:975 comparison 966:975 comparison with two structurally related analogues 966:1015 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 7 38 theme exocyclic 1344:1352 arg1 substituent 1354:1364 the exocyclic substituent 1340:1364 the exocyclic substituent 1340:1364 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 4 39 theme PC 821:822 arg1 effect 824:829 the PC effect 817:829 the PC effect of two competitive inhibitors of β-Gal 817:868 We have also evaluated the PC effect of two competitive inhibitors of β-Gal. 24737316 7 40 dep orientation 1305:1315 arg1 the 1301:1303 the 1301:1303 the 1301:1303 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 5 41 theme mechanism 934:942 arg1 view 910:913 a detailed atomic view 892:913 a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues 892:1015 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 8 42 theme action 1423:1428 arg1 mechanism 1410:1418 the mechanism 1406:1418 the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds 1406:1491 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 2 43 theme PC 333:334 arg1 approach 388:395 a newly developed molecular therapeutic approach 348:395 a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 348:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 43 theme PC 333:334 arg1 therapy 337:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 3 44 theme β-Gal 680:684 arg1 properties 639:648 the enzymological properties 621:648 the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T) 621:791 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 2 45 theme enzyme 443:448 arg1 ligands 421:427 small molecule ligands 406:427 small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 406:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 5 46 with comparison 966:975 arg1 analogues 1007:1015 two structurally related analogues 982:1015 two structurally related analogues 982:1015 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 2 47 theme developed 356:364 arg1 approach 388:395 a newly developed molecular therapeutic approach 348:395 a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome 348:590 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 2 47 theme developed 356:364 arg1 therapy 337:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy 306:343 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 6 48 theme active 1122:1127 arg1 residues 1134:1141 active site residues 1122:1141 active site residues 1122:1141 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 3 49 theme enzymological 625:637 arg1 properties 639:648 the enzymological properties 621:648 the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T) 621:791 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 3 50 from properties 639:648 arg1 patients 754:761 GM1 gangliosidosis Japanese patients 726:761 GM1 gangliosidosis Japanese patients 726:761 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 2 51 theme mutant 436:441 arg1 enzyme 443:448 the mutant enzyme 432:448 the mutant enzyme 432:448 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 1 52 dep related 223:229 arg1 diseases 140:147 autosomal recessive diseases 120:147 autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal) 120:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 52 dep related 223:229 arg1 gangliosidosis 79:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 52 dep related 223:229 arg1 disease 108:114 Morquio B disease 98:114 Morquio B disease 98:114 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 2 53 theme correct 479:485 arg1 folding 487:493 the correct folding 475:493 the correct folding 475:493 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 8 54 theme β-Gal 1433:1437 arg1 compounds 1483:1491 β-Gal selective chaperoning by newly developed PC compounds 1433:1491 β-Gal selective chaperoning by newly developed PC compounds 1433:1491 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 1 55 theme autosomal 120:128 arg1 diseases 140:147 autosomal recessive diseases 120:147 autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal) 120:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 55 theme autosomal 120:128 arg1 gangliosidosis 79:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 55 theme autosomal 120:128 arg1 disease 108:114 Morquio B disease 98:114 Morquio B disease 98:114 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 0 56 theme human 52:56 arg1 β-galactosidase 58:72 human β-galactosidase 52:72 human β-galactosidase 52:72 Structural basis of pharmacological chaperoning for human β-galactosidase. 24737316 3 57 theme representative 698:711 arg1 β-Gal 781:785 β-Gal 781:785 β-Gal 781:785 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 3 57 theme representative 698:711 arg1 β-Gal 764:768 β-Gal 764:768 β-Gal 764:768 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 3 57 theme representative 698:711 arg1 mutations 713:721 two representative mutations 694:721 two representative mutations in GM1 gangliosidosis Japanese patients 694:761 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 1 58 theme recessive 130:138 arg1 diseases 140:147 autosomal recessive diseases 120:147 autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal) 120:209 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 58 theme recessive 130:138 arg1 gangliosidosis 79:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis 75:92 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 1 58 theme recessive 130:138 arg1 disease 108:114 Morquio B disease 98:114 Morquio B disease 98:114 GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. 24737316 3 59 theme mutations 713:721 arg1 properties 639:648 the enzymological properties 621:648 the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T) 621:791 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 3 60 dep β-Gal 680:684 arg1 WT 686:687 WT 686:687 WT 686:687 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 5 61 theme atomic 903:908 arg1 view 910:913 a detailed atomic view 892:913 a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues 892:1015 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 7 62 theme mono- 1250:1254 arg1 structure 1268:1276 the mono- or bicyclic structure 1246:1276 the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent 1246:1364 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 6 63 theme site 1129:1132 arg1 residues 1134:1141 active site residues 1122:1141 active site residues 1122:1141 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 7 64 theme PC 1208:1209 arg1 potential 1211:1219 the PC potential 1204:1219 the PC potential 1204:1219 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 8 65 theme selective 1439:1447 arg1 compounds 1483:1491 β-Gal selective chaperoning by newly developed PC compounds 1433:1491 β-Gal selective chaperoning by newly developed PC compounds 1433:1491 These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds. 24737316 5 66 theme related 999:1005 arg1 analogues 1007:1015 two structurally related analogues 982:1015 two structurally related analogues 982:1015 Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. 24737316 6 67 theme hydrogen 1104:1111 arg1 bonds 1113:1117 hydrogen bonds 1104:1117 hydrogen bonds 1104:1117 All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. 24737316 2 68 theme reticulum-associated 519:538 arg1 degradation 540:550 endoplasmic reticulum-associated degradation 507:550 endoplasmic reticulum-associated degradation 507:550 Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. 24737316 3 69 theme gangliosidosis 730:743 arg1 patients 754:761 GM1 gangliosidosis Japanese patients 726:761 GM1 gangliosidosis Japanese patients 726:761 In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). 24737316 7 70 theme binding 1158:1164 arg1 affinity 1166:1173 the binding affinity 1154:1173 the binding affinity 1154:1173 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 24737316 7 71 theme bicyclic 1259:1266 arg1 structure 1268:1276 the mono- or bicyclic structure 1246:1276 the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent 1246:1364 Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. 34074792 4 0 theme NMR 976:978 arg1 methods 980:986 NMR methods 976:986 NMR methods for studies of condensed IDR states 976:1022 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 7 1 theme detailed 1514:1521 arg1 information 1535:1545 detailed interaction information 1514:1545 detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates 1514:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 2 2 theme large-scale 651:661 arg1 states 674:679 these highly dynamic large-scale associated states 630:679 these highly dynamic large-scale associated states 630:679 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 4 3 theme messenger 899:907 arg1 RNA 909:911 messenger RNA 899:911 messenger RNA transport granules 899:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 2 4 theme significant 378:388 arg1 insights 390:397 significant insights 378:397 significant insights into IDR phase behavior 378:421 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 4 5 located found 859:863 arg2 protein 851:857 an RNA-binding protein 836:857 an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules 836:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 5 located found 859:863 arg1 granules 923:930 messenger RNA transport granules 899:930 messenger RNA transport granules 899:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 5 located found 859:863 arg2 CAPRIN1 827:833 CAPRIN1 827:833 CAPRIN1 827:833 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 5 located found 859:863 arg1 granules 875:882 stress granules 868:882 stress granules 868:882 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 5 located found 859:863 arg1 bodies 887:892 P bodies 885:892 P bodies 885:892 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 6 theme transport 913:921 arg1 granules 923:930 messenger RNA transport granules 899:930 messenger RNA transport granules 899:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 3 7 theme dynamic 720:726 arg1 nature 740:745 the dynamic and viscous nature 716:745 the dynamic and viscous nature of condensed IDRs 716:763 NMR can be a valuable method, but the dynamic and viscous nature of condensed IDRs presents challenges. 34074792 2 8 theme dynamic 643:649 arg1 states 674:679 these highly dynamic large-scale associated states 630:679 these highly dynamic large-scale associated states 630:679 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 4 9 dep interactions 1049:1060 arg1 modulating 1074:1083 modulating 1074:1083 modulating phase separation 1074:1100 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 9 dep interactions 1049:1060 arg1 driving 1062:1068 driving 1062:1068 driving 1062:1068 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 1 10 theme biological 148:157 arg1 function 159:166 biological function 148:166 biological function 148:166 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 0 11 theme condensed 80:88 arg1 phase 90:94 a CAPRIN1 condensed phase 70:94 a CAPRIN1 condensed phase 70:94 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 6 12 theme critical 1290:1297 arg1 sequences 1299:1307 critical sequences 1290:1307 critical sequences for phase separation 1290:1328 We also quantify specific side-chain and backbone interactions within condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress. 34074792 6 13 theme condensed 1260:1268 arg1 CAPRIN1 1270:1276 condensed CAPRIN1 1260:1276 condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress 1260:1412 We also quantify specific side-chain and backbone interactions within condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress. 34074792 5 14 with interactions 1119:1130 arg1 CAPRIN1 1137:1143 CAPRIN1 1137:1143 CAPRIN1 1137:1143 We identify ATP interactions with CAPRIN1 that can enhance or reduce phase separation. 34074792 0 15 theme CAPRIN1 72:78 arg1 phase 90:94 a CAPRIN1 condensed phase 70:94 a CAPRIN1 condensed phase 70:94 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 3 16 theme IDRs 760:763 arg1 nature 740:745 the dynamic and viscous nature 716:745 the dynamic and viscous nature of condensed IDRs 716:763 NMR can be a valuable method, but the dynamic and viscous nature of condensed IDRs presents challenges. 34074792 4 17 theme P 885:885 arg1 bodies 887:892 P bodies 885:892 P bodies 885:892 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 18 theme C-terminal 796:805 arg1 IDR 807:809 the C-terminal IDR 792:809 the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules 792:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 18 theme C-terminal 796:805 arg1 709 819:821 709 819:821 709 819:821 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 19 theme methods 980:986 arg1 variety 965:971 a variety 963:971 a variety of NMR methods for studies of condensed IDR states 963:1022 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 19 theme methods 980:986 arg1 methods 980:986 NMR methods 976:986 NMR methods for studies of condensed IDR states 976:1022 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 7 20 theme expanded 1420:1427 arg1 toolkit 1433:1439 This expanded NMR toolkit 1415:1439 This expanded NMR toolkit that has been developed for characterizing IDR condensates 1415:1498 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 7 21 theme IDR 1484:1486 arg1 condensates 1488:1498 IDR condensates 1484:1498 IDR condensates 1484:1498 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 7 22 theme phase 1622:1626 arg1 separation 1628:1637 phase separation 1622:1637 phase separation 1622:1637 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 2 23 theme critical 441:448 arg1 interactions 450:461 the critical interactions 437:461 the critical interactions that govern condensation with atomic resolution through experiment 437:528 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 4 24 dep 709 819:821 arg1 to 816:817 to 816:817 to 816:817 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 0 25 theme phase 90:94 arg1 studies 59:65 NMR studies 55:65 NMR studies of a CAPRIN1 condensed phase 55:94 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 4 26 theme condensed 1003:1011 arg1 states 1017:1022 condensed IDR states 1003:1022 condensed IDR states 1003:1022 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 7 27 theme significant 1645:1655 arg1 applications 1674:1685 significant potential future applications 1645:1685 significant potential future applications 1645:1685 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 0 28 theme hot 12:14 arg1 spots 16:20 Interaction hot spots 0:20 Interaction hot spots for phase separation 0:41 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 3 29 theme condensed 750:758 arg1 IDRs 760:763 condensed IDRs 750:763 condensed IDRs 750:763 NMR can be a valuable method, but the dynamic and viscous nature of condensed IDRs presents challenges. 34074792 4 30 theme stress 868:873 arg1 granules 875:882 stress granules 868:882 stress granules 868:882 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 2 31 theme computational 323:335 arg1 approaches 353:362 computational and theoretical approaches 323:362 computational and theoretical approaches 323:362 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 2 32 theme phase 408:412 arg1 behavior 414:421 IDR phase behavior 404:421 IDR phase behavior 404:421 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 0 33 theme Interaction 0:10 arg1 spots 16:20 Interaction hot spots 0:20 Interaction hot spots for phase separation 0:41 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 7 34 theme dynamic 1701:1707 arg1 relationships 1728:1740 dynamic structure-function relationships 1701:1740 dynamic structure-function relationships in other biological condensates 1701:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 2 35 theme associated 663:672 arg1 states 674:679 these highly dynamic large-scale associated states 630:679 these highly dynamic large-scale associated states 630:679 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 7 36 theme NMR 1429:1431 arg1 toolkit 1433:1439 This expanded NMR toolkit 1415:1439 This expanded NMR toolkit that has been developed for characterizing IDR condensates 1415:1498 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 7 37 theme other 1745:1749 arg1 condensates 1762:1772 other biological condensates 1745:1772 other biological condensates 1745:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 2 38 theme IDR 404:406 arg1 behavior 414:421 IDR phase behavior 404:421 IDR phase behavior 404:421 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 5 39 theme phase 1172:1176 arg1 separation 1178:1187 phase separation 1172:1187 phase separation 1172:1187 We identify ATP interactions with CAPRIN1 that can enhance or reduce phase separation. 34074792 2 40 theme structural 593:602 arg1 tools 615:619 standard structural biological tools 584:619 standard structural biological tools 584:619 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 1 41 from role 101:104 arg1 assembly 278:285 their assembly 272:285 their assembly 272:285 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 6 42 theme cell 1392:1395 arg1 cycle 1397:1401 cell cycle 1392:1401 cell cycle 1392:1401 We also quantify specific side-chain and backbone interactions within condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress. 34074792 7 43 theme biological 1751:1760 arg1 condensates 1762:1772 other biological condensates 1745:1772 other biological condensates 1745:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 4 44 theme states 1017:1022 arg1 studies 992:998 studies 992:998 studies of condensed IDR states 992:1022 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 2 45 theme standard 584:591 arg1 tools 615:619 standard structural biological tools 584:619 standard structural biological tools 584:619 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 1 46 theme dynamic 190:196 arg1 interactions 198:209 dynamic interactions 190:209 dynamic interactions involving intrinsically disordered protein regions (IDRs) 190:267 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 7 47 theme structure-function 1709:1726 arg1 relationships 1728:1740 dynamic structure-function relationships 1701:1740 dynamic structure-function relationships in other biological condensates 1701:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 3 48 theme valuable 695:702 arg1 method 704:709 a valuable method 693:709 a valuable method 693:709 NMR can be a valuable method, but the dynamic and viscous nature of condensed IDRs presents challenges. 34074792 3 48 theme valuable 695:702 arg1 NMR 682:684 NMR 682:684 NMR 682:684 NMR can be a valuable method, but the dynamic and viscous nature of condensed IDRs presents challenges. 34074792 7 49 from relationships 1728:1740 arg1 condensates 1762:1772 other biological condensates 1745:1772 other biological condensates 1745:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 1 50 theme interactions 198:209 arg1 importance 176:185 the importance 172:185 the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly 172:285 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 1 50 theme interactions 198:209 arg1 role 101:104 The role 97:104 The role of biomolecular condensates in regulating biological function 97:166 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 0 51 theme phase 26:30 arg1 separation 32:41 phase separation 26:41 phase separation 26:41 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 7 52 theme future 1667:1672 arg1 applications 1674:1685 significant potential future applications 1645:1685 significant potential future applications 1645:1685 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 2 53 theme applicability 567:579 arg1 lack 559:562 the lack 555:562 the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states 555:679 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 4 54 theme CAPRIN1 827:833 arg1 IDR 807:809 the C-terminal IDR 792:809 the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules 792:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 54 theme CAPRIN1 827:833 arg1 709 819:821 709 819:821 709 819:821 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 6 55 theme phase 1313:1317 arg1 separation 1319:1328 phase separation 1313:1328 phase separation 1313:1328 We also quantify specific side-chain and backbone interactions within condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress. 34074792 7 56 theme CAPRIN1 1574:1580 arg1 biology 1582:1588 CAPRIN1 biology 1574:1588 CAPRIN1 biology 1574:1588 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 4 57 theme RNA-binding 839:849 arg1 protein 851:857 an RNA-binding protein 836:857 an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules 836:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 57 theme RNA-binding 839:849 arg1 CAPRIN1 827:833 CAPRIN1 827:833 CAPRIN1 827:833 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 4 58 theme RNA 909:911 arg1 granules 923:930 messenger RNA transport granules 899:930 messenger RNA transport granules 899:930 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 2 59 theme atomic 493:498 arg1 resolution 500:509 atomic resolution 493:509 atomic resolution 493:509 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 6 60 theme backbone 1231:1238 arg1 interactions 1240:1251 specific side-chain and backbone interactions 1207:1251 interactions 1240:1251 We also quantify specific side-chain and backbone interactions within condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress. 34074792 5 61 theme ATP 1115:1117 arg1 interactions 1119:1130 ATP interactions 1115:1130 ATP interactions with CAPRIN1 that can enhance or reduce phase separation 1115:1187 We identify ATP interactions with CAPRIN1 that can enhance or reduce phase separation. 34074792 7 62 theme relevant 1547:1554 arg1 information 1535:1545 detailed interaction information 1514:1545 detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates 1514:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 7 63 theme potential 1657:1665 arg1 applications 1674:1685 significant potential future applications 1645:1685 significant potential future applications 1645:1685 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 2 64 theme tools 615:619 arg1 applicability 567:579 applicability 567:579 applicability of standard structural biological tools 567:619 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 6 65 theme specific 1207:1214 arg1 side-chain 1216:1225 specific side-chain and backbone interactions 1207:1251 side-chain 1216:1225 We also quantify specific side-chain and backbone interactions within condensed CAPRIN1 that define critical sequences for phase separation and that are reduced by O-GlcNAcylation known to occur during cell cycle and stress. 34074792 2 66 with condensation 475:486 arg1 resolution 500:509 atomic resolution 493:509 atomic resolution 493:509 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 2 67 theme theoretical 341:351 arg1 approaches 353:362 computational and theoretical approaches 323:362 computational and theoretical approaches 323:362 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 2 68 theme biological 604:613 arg1 tools 615:619 standard structural biological tools 584:619 standard structural biological tools 584:619 While computational and theoretical approaches have provided significant insights into IDR phase behavior, establishing the critical interactions that govern condensation with atomic resolution through experiment is more difficult, given the lack of applicability of standard structural biological tools to study these highly dynamic large-scale associated states. 34074792 7 69 theme interaction 1523:1533 arg1 information 1535:1545 detailed interaction information 1514:1545 detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates 1514:1772 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 1 70 theme biomolecular 109:120 arg1 condensates 122:132 biomolecular condensates 109:132 biomolecular condensates 109:132 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 4 71 theme phase 1085:1089 arg1 separation 1091:1100 phase separation 1085:1100 phase separation 1085:1100 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 1 72 theme disordered 235:244 arg1 IDRs 263:266 IDRs 263:266 IDRs 263:266 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 1 72 theme disordered 235:244 arg1 regions 254:260 intrinsically disordered protein regions 221:260 intrinsically disordered protein regions (IDRs) 221:267 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 7 73 theme general 1604:1610 arg1 models 1612:1617 general models 1604:1617 general models 1604:1617 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 34074792 1 74 from importance 176:185 arg1 assembly 278:285 their assembly 272:285 their assembly 272:285 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 1 75 theme condensates 122:132 arg1 importance 176:185 the importance 172:185 the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly 172:285 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 1 75 theme condensates 122:132 arg1 role 101:104 The role 97:104 The role of biomolecular condensates in regulating biological function 97:166 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 1 76 theme protein 246:252 arg1 IDRs 263:266 IDRs 263:266 IDRs 263:266 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 1 76 theme protein 246:252 arg1 regions 254:260 intrinsically disordered protein regions 221:260 intrinsically disordered protein regions (IDRs) 221:267 The role of biomolecular condensates in regulating biological function and the importance of dynamic interactions involving intrinsically disordered protein regions (IDRs) in their assembly are increasingly appreciated. 34074792 0 77 theme NMR 55:57 arg1 studies 59:65 NMR studies 55:65 NMR studies of a CAPRIN1 condensed phase 55:94 Interaction hot spots for phase separation revealed by NMR studies of a CAPRIN1 condensed phase. 34074792 3 78 theme viscous 732:738 arg1 nature 740:745 the dynamic and viscous nature 716:745 the dynamic and viscous nature of condensed IDRs 716:763 NMR can be a valuable method, but the dynamic and viscous nature of condensed IDRs presents challenges. 34074792 4 79 theme IDR 1013:1015 arg1 states 1017:1022 condensed IDR states 1003:1022 condensed IDR states 1003:1022 Using the C-terminal IDR (607 to 709) of CAPRIN1, an RNA-binding protein found in stress granules, P bodies, and messenger RNA transport granules, we have developed and applied a variety of NMR methods for studies of condensed IDR states to provide insights into interactions driving and modulating phase separation. 34074792 7 80 theme separation 1628:1637 arg1 models 1612:1617 general models 1604:1617 general models 1604:1617 This expanded NMR toolkit that has been developed for characterizing IDR condensates has generated detailed interaction information relevant for understanding CAPRIN1 biology and informing general models of phase separation, with significant potential future applications to illuminate dynamic structure-function relationships in other biological condensates. 2129367 4 0 theme S-aminoethylated 452:467 arg1 VII 476:478 the S-aminoethylated factor VII 448:478 the S-aminoethylated factor VII 448:478 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 3 1 theme growth 378:383 arg1 VII 414:416 factor VII 407:416 factor VII 407:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 3 1 theme growth 378:383 arg1 domain 397:402 the first epidermal growth factor-like domain 358:402 the first epidermal growth factor-like domain of factor VII 358:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 9 2 theme Xyl2 1236:1239 arg1 structure 1250:1258 a (Xyl2)-Glc-Ser structure 1233:1258 a (Xyl2)-Glc-Ser structure in glycoproteins 1233:1275 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 10 3 attach presence 1299:1306 arg1 IX 1362:1363 IX 1362:1363 IX 1362:1363 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 10 3 attach presence 1299:1306 arg1 Z 1377:1377 protein Z 1369:1377 protein Z 1369:1377 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 10 3 attach presence 1299:1306 arg2 structure 1336:1344 the unique trisaccharide structure 1311:1344 the unique trisaccharide structure 1311:1344 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 10 3 attach presence 1299:1306 arg1 VII 1357:1359 factors VII 1349:1359 factors VII 1349:1359 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 9 4 theme -Glc-Ser 1241:1248 arg1 structure 1250:1258 a (Xyl2)-Glc-Ser structure 1233:1258 a (Xyl2)-Glc-Ser structure in glycoproteins 1233:1275 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 10 5 from role 1417:1420 arg1 pathway 1443:1449 the tissue factor pathway 1425:1449 the tissue factor pathway 1425:1449 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 10 6 theme protein 1369:1375 arg1 Z 1377:1377 protein Z 1369:1377 protein Z 1369:1377 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 3 7 theme factor 407:412 arg1 VII 414:416 factor VII 407:416 factor VII 407:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 5 8 theme glucose 727:733 arg1 mol 741:743 2 mol 739:743 2 mol of xylose 739:753 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 5 8 theme glucose 727:733 arg1 mol 720:722 1 mol 718:722 1 mol of glucose 718:733 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 0 9 attach linked 32:37 arg2 chain 26:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 9 attach linked 32:37 arg1 residue 51:57 a serine residue 42:57 a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 42:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 10 theme amino 171:175 arg1 sequence 182:189 the complete amino acid sequence 158:189 the complete amino acid sequence of bovine factor VII 158:210 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 8 11 theme Xyl2 1077:1080 arg1 structure 1090:1098 a (Xyl2)Glc-Ser structure 1074:1098 a (Xyl2)Glc-Ser structure 1074:1098 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 6 12 theme bombardment 825:835 arg1 analysis 856:863 high-resolution fast atom bombardment mass spectrometric analysis 799:863 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 0 13 theme clotting 91:98 arg1 VII 108:110 VII 108:110 VII 108:110 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 13 theme clotting 91:98 arg1 protein 123:129 protein 123:129 protein 123:129 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 13 theme clotting 91:98 arg1 factors 100:106 clotting factors 91:106 clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 91:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 13 theme clotting 91:98 arg1 IX 116:117 IX 116:117 IX 116:117 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 8 14 theme Glc-Ser 1082:1088 arg1 structure 1090:1098 a (Xyl2)Glc-Ser structure 1074:1098 a (Xyl2)Glc-Ser structure 1074:1098 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 6 15 theme fast 815:818 arg1 bombardment 825:835 high-resolution fast atom bombardment 799:835 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 8 16 theme Similar 1134:1140 arg1 results 1142:1148 Similar results 1134:1148 Similar results 1134:1148 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 9 17 gly glycoproteins 1263:1275 arg1 glycoproteins 1263:1275 glycoproteins 1263:1275 glycoproteins 1263:1275 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 0 18 dep IX 116:117 arg1 1988 232:235 1988 232:235 1988 232:235 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 18 dep IX 116:117 arg1 Z. 131:132 Z. 131:132 Z. 131:132 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 10 19 from presence 1299:1306 arg1 IX 1362:1363 IX 1362:1363 IX 1362:1363 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 10 19 from presence 1299:1306 arg1 Z 1377:1377 protein Z 1369:1377 protein Z 1369:1377 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 10 19 from presence 1299:1306 arg1 VII 1357:1359 factors VII 1349:1359 factors VII 1349:1359 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 5 20 theme factor 632:637 arg1 IX 639:640 factor IX 632:640 factor IX 632:640 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 6 21 theme pentapeptide 872:883 arg1 analysis 856:863 high-resolution fast atom bombardment mass spectrometric analysis 799:863 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 0 22 theme first 66:70 arg1 domain 81:86 the first EGF-like domain 62:86 the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 62:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 22 theme first 66:70 arg1 factors 100:106 clotting factors 91:106 clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 91:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 23 theme factor 201:206 arg1 VII 208:210 bovine factor VII 194:210 bovine factor VII 194:210 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 8 24 dep Z. 1131:1132 arg1 obtained 1155:1162 obtained 1155:1162 were obtained for human factors VII, IX and protein Z 1150:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 6 25 theme spectrometric 842:854 arg1 analysis 856:863 high-resolution fast atom bombardment mass spectrometric analysis 799:863 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 5 26 contain containing 611:620 arg2 Ser-53 622:627 Ser-53 622:627 Ser-53 of factor IX 622:640 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 5 26 contain containing 611:620 arg1 pentapeptide 598:609 a pentapeptide 596:609 a pentapeptide containing Ser-53 of factor IX 596:640 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 10 27 theme unique 1315:1320 arg1 structure 1336:1344 the unique trisaccharide structure 1311:1344 the unique trisaccharide structure 1311:1344 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 0 28 theme trisaccharide 6:18 arg1 chain 26:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 3 29 theme studies 293:299 arg1 course 279:284 the course 275:284 the course of the studies 275:299 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 3 30 theme serine 322:327 arg1 derivative 329:338 an unknown serine derivative 311:338 an unknown serine derivative 311:338 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 5 31 theme Z. 654:655 arg1 analysis 673:680 protein Z. Component sugar analysis 646:680 protein Z. Component sugar analysis 646:680 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 10 32 theme factors 1349:1355 arg1 VII 1357:1359 factors VII 1349:1359 factors VII 1349:1359 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 0 33 theme serine 44:49 arg1 residue 51:57 a serine residue 42:57 a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 42:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 5 34 theme sugar 667:671 arg1 analysis 673:680 protein Z. Component sugar analysis 646:680 protein Z. Component sugar analysis 646:680 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 8 35 from existence 1061:1069 arg1 protein 1123:1129 protein 1123:1129 protein 1123:1129 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 35 from existence 1061:1069 arg1 IX 1116:1117 IX 1116:1117 IX 1116:1117 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 35 from existence 1061:1069 arg1 VII 1111:1113 factors VII 1103:1113 factors VII 1103:1113 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 10 36 theme structure 1336:1344 arg1 presence 1299:1306 The presence 1295:1306 The presence of the unique trisaccharide structure in factors VII, IX and protein Z 1295:1377 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 9 37 theme first 1217:1221 arg1 This 1205:1208 This 1205:1208 This 1205:1208 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 9 37 theme first 1217:1221 arg1 report 1223:1228 the first report 1213:1228 the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge 1213:1292 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 9 38 from report 1223:1228 arg1 glycoproteins 1263:1275 glycoproteins 1263:1275 glycoproteins 1263:1275 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 9 39 from glycoproteins 1263:1275 arg1 This 1205:1208 This 1205:1208 This 1205:1208 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 9 39 from glycoproteins 1263:1275 arg1 report 1223:1228 the first report 1213:1228 the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge 1213:1292 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 3 40 theme factor-like 385:395 arg1 VII 414:416 factor VII 407:416 factor VII 407:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 3 40 theme factor-like 385:395 arg1 domain 397:402 the first epidermal growth factor-like domain 358:402 the first epidermal growth factor-like domain of factor VII 358:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 9 41 from structure 1250:1258 arg1 glycoproteins 1263:1275 glycoproteins 1263:1275 glycoproteins 1263:1275 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 3 42 theme epidermal 368:376 arg1 VII 414:416 factor VII 407:416 factor VII 407:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 3 42 theme epidermal 368:376 arg1 domain 397:402 the first epidermal growth factor-like domain 358:402 the first epidermal growth factor-like domain of factor VII 358:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 6 43 theme sugar 761:765 arg1 component 767:775 This sugar component 756:775 This sugar component 756:775 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 0 44 theme acid 177:180 arg1 sequence 182:189 the complete amino acid sequence 158:189 the complete amino acid sequence of bovine factor VII 158:210 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 7 45 theme reducing 981:988 arg1 end 990:992 its reducing end 977:992 its reducing end 977:992 The trisaccharide was released from the peptides by means of beta-elimination reaction and its reducing end was identified as pyridylamino-glucose. 2129367 7 45 theme reducing 981:988 arg1 pyridylamino-glucose 1012:1031 pyridylamino-glucose 1012:1031 pyridylamino-glucose 1012:1031 The trisaccharide was released from the peptides by means of beta-elimination reaction and its reducing end was identified as pyridylamino-glucose. 2129367 3 46 theme VII 414:416 arg1 VII 414:416 factor VII 407:416 factor VII 407:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 3 46 theme VII 414:416 arg1 domain 397:402 the first epidermal growth factor-like domain 358:402 the first epidermal growth factor-like domain of factor VII 358:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 5 47 theme xylose 748:753 arg1 mol 741:743 2 mol 739:743 2 mol of xylose 739:753 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 5 47 theme xylose 748:753 arg1 mol 720:722 1 mol 718:722 1 mol of glucose 718:733 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 4 48 theme factor 469:474 arg1 VII 476:478 the S-aminoethylated factor VII 448:478 the S-aminoethylated factor VII 448:478 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 0 49 theme complete 162:169 arg1 sequence 182:189 the complete amino acid sequence 158:189 the complete amino acid sequence of bovine factor VII 158:210 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 4 50 attach isolated 434:441 arg2 pentapeptide 421:432 A pentapeptide 419:432 A pentapeptide isolated from the S-aminoethylated factor VII 419:478 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 4 50 attach isolated 434:441 arg1 VII 476:478 the S-aminoethylated factor VII 448:478 the S-aminoethylated factor VII 448:478 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 9 51 theme structure 1250:1258 arg1 This 1205:1208 This 1205:1208 This 1205:1208 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 9 51 theme structure 1250:1258 arg1 report 1223:1228 the first report 1213:1228 the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge 1213:1292 This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. 2129367 7 52 attach released 908:915 arg2 trisaccharide 890:902 The trisaccharide 886:902 The trisaccharide 886:902 The trisaccharide was released from the peptides by means of beta-elimination reaction and its reducing end was identified as pyridylamino-glucose. 2129367 7 52 attach released 908:915 arg1 peptides 926:933 the peptides 922:933 the peptides 922:933 The trisaccharide was released from the peptides by means of beta-elimination reaction and its reducing end was identified as pyridylamino-glucose. 2129367 8 53 theme factors 1103:1109 arg1 VII 1111:1113 factors VII 1103:1113 factors VII 1103:1113 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 10 54 theme biological 1406:1415 arg1 role 1417:1420 its biological role 1402:1420 its biological role in the tissue factor pathway 1402:1449 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 7 55 theme beta-elimination 947:962 arg1 reaction 964:971 beta-elimination reaction 947:971 beta-elimination reaction 947:971 The trisaccharide was released from the peptides by means of beta-elimination reaction and its reducing end was identified as pyridylamino-glucose. 2129367 6 56 theme mass 837:840 arg1 analysis 856:863 high-resolution fast atom bombardment mass spectrometric analysis 799:863 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 4 57 theme gas-phase 535:543 arg1 sequencer 545:553 a gas-phase sequencer 533:553 a gas-phase sequencer 533:553 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 0 58 theme factors 100:106 arg1 domain 81:86 the first EGF-like domain 62:86 the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 62:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 58 theme factors 100:106 arg1 factors 100:106 clotting factors 91:106 clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 91:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 59 dep determined 147:156 arg1 H. 221:222 H. 221:222 H. 221:222 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 6 60 theme atom 820:823 arg1 bombardment 825:835 high-resolution fast atom bombardment 799:835 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 10 61 theme tissue 1429:1434 arg1 pathway 1443:1449 the tissue factor pathway 1425:1449 the tissue factor pathway 1425:1449 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 6 62 theme high-resolution 799:813 arg1 bombardment 825:835 high-resolution fast atom bombardment 799:835 high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide 799:883 This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. 2129367 0 63 from residue 51:57 arg1 domain 81:86 the first EGF-like domain 62:86 the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 62:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 63 from residue 51:57 arg1 factors 100:106 clotting factors 91:106 clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 91:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 64 dep Z. 131:132 arg1 Biol 241:244 Biol 241:244 Biol 241:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 64 dep Z. 131:132 arg1 determined 147:156 determined 147:156 determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al 147:228 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 8 65 theme structure 1090:1098 arg1 existence 1061:1069 the existence 1057:1069 the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z 1057:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 5 66 theme same 560:563 arg1 results 565:571 The same results 556:571 The same results 556:571 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 4 67 contain contained 480:488 arg1 pentapeptide 421:432 A pentapeptide 419:432 A pentapeptide isolated from the S-aminoethylated factor VII 419:478 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 4 67 contain contained 480:488 arg2 Ser-52 490:495 Ser-52 490:495 Ser-52 490:495 A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. 2129367 8 68 theme human 1168:1172 arg1 factors 1174:1180 human factors 1168:1180 human factors VII, IX and protein Z 1168:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 68 theme human 1168:1172 arg1 IX 1187:1188 IX 1187:1188 IX 1187:1188 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 68 theme human 1168:1172 arg1 Z 1202:1202 protein Z 1194:1202 protein Z 1194:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 68 theme human 1168:1172 arg1 VII 1182:1184 VII 1182:1184 VII 1182:1184 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 5 69 theme IX 639:640 arg1 Ser-53 622:627 Ser-53 622:627 Ser-53 of factor IX 622:640 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 0 70 dep factors 100:106 arg1 VII 108:110 VII 108:110 VII 108:110 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 70 dep factors 100:106 arg1 factors 100:106 clotting factors 91:106 clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 91:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 70 dep factors 100:106 arg1 IX 116:117 IX 116:117 IX 116:117 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 71 theme EGF-like 72:79 arg1 domain 81:86 the first EGF-like domain 62:86 the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 62:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 71 theme EGF-like 72:79 arg1 factors 100:106 clotting factors 91:106 clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol 91:244 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 2 72 dep 263 253:255 arg1 14868-14877 258:268 14868-14877 258:268 14868-14877 258:268 263, 14868-14877). 2129367 0 73 theme bovine 194:199 arg1 VII 208:210 bovine factor VII 194:210 bovine factor VII 194:210 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 8 74 theme protein 1194:1200 arg1 factors 1174:1180 human factors 1168:1180 human factors VII, IX and protein Z 1168:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 74 theme protein 1194:1200 arg1 Z 1202:1202 protein Z 1194:1202 protein Z 1194:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 3 75 theme unknown 314:320 arg1 derivative 329:338 an unknown serine derivative 311:338 an unknown serine derivative 311:338 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 0 76 theme new 2:4 arg1 chain 26:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 0 77 theme VII 208:210 arg1 sequence 182:189 the complete amino acid sequence 158:189 the complete amino acid sequence of bovine factor VII 158:210 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 5 78 contain contained 708:716 arg2 mol 741:743 2 mol 739:743 2 mol of xylose 739:753 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 5 78 contain contained 708:716 arg1 peptide 700:706 the peptide 696:706 the peptide 696:706 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 5 78 contain contained 708:716 arg2 mol 720:722 1 mol 718:722 1 mol of glucose 718:733 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 10 79 theme factor 1436:1441 arg1 pathway 1443:1449 the tissue factor pathway 1425:1449 the tissue factor pathway 1425:1449 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 2129367 0 80 theme sugar 20:24 arg1 chain 26:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain 0:30 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 8 81 dep VII 1111:1113 arg1 Z. 1131:1132 Z. 1131:1132 Z. 1131:1132 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 5 82 theme protein 646:652 arg1 analysis 673:680 protein Z. Component sugar analysis 646:680 protein Z. Component sugar analysis 646:680 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 5 83 theme Component 657:665 arg1 analysis 673:680 protein Z. Component sugar analysis 646:680 protein Z. Component sugar analysis 646:680 The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. 2129367 8 84 dep factors 1174:1180 arg1 factors 1174:1180 human factors 1168:1180 human factors VII, IX and protein Z 1168:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 84 dep factors 1174:1180 arg1 IX 1187:1188 IX 1187:1188 IX 1187:1188 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 84 dep factors 1174:1180 arg1 Z 1202:1202 protein Z 1194:1202 protein Z 1194:1202 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 8 84 dep factors 1174:1180 arg1 VII 1182:1184 VII 1182:1184 VII 1182:1184 These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. 2129367 3 85 theme first 362:366 arg1 VII 414:416 factor VII 407:416 factor VII 407:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 3 85 theme first 362:366 arg1 domain 397:402 the first epidermal growth factor-like domain 358:402 the first epidermal growth factor-like domain of factor VII 358:416 In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. 2129367 0 86 dep H. 221:222 arg1 al 227:228 et al 224:228 Takeya, H. et al 213:228 A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z. Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. 2129367 10 87 theme trisaccharide 1322:1334 arg1 structure 1336:1344 the unique trisaccharide structure 1311:1344 the unique trisaccharide structure 1311:1344 The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway. 21949356 7 0 from glucose 1043:1049 arg1 sequence 1091:1098 the O-glucose consensus sequence 1067:1098 the O-glucose consensus sequence ( ) of factor VII EGF repeat 1067:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 3 1 dep exist 382:386 arg1 one 389:391 one 389:391 one 389:391 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 3 1 dep exist 382:386 arg1 others 458:463 two others 454:463 two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2) 454:543 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 11 2 used used 1721:1724 arg2 substrate 1683:1691 donor substrate 1677:1691 donor substrate (UDP-glucose or UDP-Xyl) 1677:1716 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 1 3 theme signaling 157:165 arg1 loss 143:146 a temperature-sensitive loss 119:146 a temperature-sensitive loss of Notch signaling in Drosophila 119:179 Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. 21949356 9 4 from glycans 1370:1376 arg1 cells 1381:1385 cells 1381:1385 cells 1381:1385 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 9 5 theme mouse 1250:1254 arg1 Notch2 1256:1261 mouse Notch2 1250:1261 mouse Notch2 1250:1261 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 7 6 theme factor 1107:1112 arg1 repeat 1122:1127 factor VII EGF repeat 1107:1127 factor VII EGF repeat 1107:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 3 7 theme lower 470:474 arg1 degrees 476:482 lower degrees 470:482 lower degrees of identity 470:494 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 9 8 mod modified 1330:1337 arg1 EGF16 1241:1245 EGF16 1241:1245 EGF16 1241:1245 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 9 8 mod modified 1330:1337 arg3 O-Xyl 1351:1355 O-Xyl 1351:1355 O-Xyl 1351:1355 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 9 8 mod modified 1330:1337 arg3 glycans 1370:1376 O-glucose glycans 1360:1376 O-glucose glycans in cells 1360:1385 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 9 9 theme Notch2 1256:1261 arg1 EGF16 1241:1245 EGF16 1241:1245 EGF16 1241:1245 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 3 10 theme Rumi 366:369 arg1 homologues 371:380 three potential Rumi homologues 350:380 three potential Rumi homologues 350:380 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 4 11 dep mouse 569:573 arg1 Rumi 585:588 Rumi 585:588 Rumi 585:588 Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDP-glucose to an EGF repeat from human factor VII. 21949356 2 12 theme protein 291:297 arg1 domain 264:269 a CAP10 domain 256:269 a CAP10 domain that functions as a protein O-glucosyltransferase 256:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 12 theme protein 291:297 arg1 O-glucosyltransferase 299:319 a protein O-glucosyltransferase 289:319 a protein O-glucosyltransferase 289:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 9 13 theme O-glucose 1360:1368 arg1 glycans 1370:1376 O-glucose glycans 1360:1376 O-glucose glycans in cells 1360:1385 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 10 14 theme second 1404:1409 arg1 S590A 1419:1423 S590A 1419:1423 S590A 1419:1423 Mutation of the second serine (S590A) causes a loss of O-Xyl but not O-glucose at this site. 21949356 10 14 theme second 1404:1409 arg1 serine 1411:1416 the second serine 1400:1416 the second serine (S590A) 1400:1424 Mutation of the second serine (S590A) causes a loss of O-Xyl but not O-glucose at this site. 21949356 11 15 dep substrate 1683:1691 arg1 UDP-Xyl 1709:1715 UDP-Xyl 1709:1715 UDP-Xyl 1709:1715 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 11 15 dep substrate 1683:1691 arg1 UDP-glucose 1694:1704 UDP-glucose 1694:1704 UDP-glucose 1694:1704 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 9 16 contain has 1270:1272 arg1 EGF16 1241:1245 EGF16 1241:1245 EGF16 1241:1245 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 9 16 contain has 1270:1272 arg2 motif 1285:1289 a diserine motif 1274:1289 a diserine motif in the consensus sequence 1274:1315 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 4 17 theme EGF 669:671 arg1 repeat 673:678 an EGF repeat 666:678 an EGF repeat 666:678 Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDP-glucose to an EGF repeat from human factor VII. 21949356 9 18 from O-Xyl 1351:1355 arg1 cells 1381:1385 cells 1381:1385 cells 1381:1385 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 11 19 theme EGF 1636:1638 arg1 repeat 1640:1645 the recipient EGF repeat 1622:1645 the recipient EGF repeat 1622:1645 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 7 20 theme consensus 1081:1089 arg1 sequence 1091:1098 the O-glucose consensus sequence 1067:1098 the O-glucose consensus sequence ( ) of factor VII EGF repeat 1067:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 11 21 theme repeat 1640:1645 arg1 sequences 1609:1617 amino acid sequences 1598:1617 amino acid sequences of the recipient EGF repeat 1598:1645 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 6 22 theme protein 982:988 arg1 activity 1011:1018 significant protein O-xylosyltransferase activity 970:1018 significant protein O-xylosyltransferase activity 970:1018 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 5 23 from phenotypes 770:779 arg1 clones 800:805 Drosophila rumi clones 784:805 Drosophila rumi clones 784:805 Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. 21949356 3 24 theme identity 487:494 arg1 degrees 476:482 lower degrees 470:482 lower degrees of identity 470:494 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 9 25 theme diserine 1276:1283 arg1 motif 1285:1289 a diserine motif 1274:1289 a diserine motif in the consensus sequence 1274:1315 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 3 26 theme Drosophila 427:436 arg1 Rumi 438:441 Drosophila Rumi 427:441 Drosophila Rumi (52%) 427:447 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 3 26 theme Drosophila 427:436 arg1 % 446:446 52% 444:446 52% 444:446 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 3 27 theme potential 356:364 arg1 homologues 371:380 three potential Rumi homologues 350:380 three potential Rumi homologues 350:380 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 6 28 theme O-glucosyltransferase 891:911 arg1 activity 913:920 protein O-glucosyltransferase activity 883:920 protein O-glucosyltransferase activity 883:920 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 6 29 theme significant 970:980 arg1 activity 1011:1018 significant protein O-xylosyltransferase activity 970:1018 significant protein O-xylosyltransferase activity 970:1018 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 10 30 theme serine 1411:1416 arg1 Mutation 1388:1395 Mutation 1388:1395 Mutation of the second serine (S590A) 1388:1424 Mutation of the second serine (S590A) causes a loss of O-Xyl but not O-glucose at this site. 21949356 6 31 theme enzymes 844:850 arg1 characterization 815:830 characterization 815:830 characterization of the Rumi enzymes 815:850 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 11 32 theme recipient 1626:1634 arg1 repeat 1640:1645 the recipient EGF repeat 1622:1645 the recipient EGF repeat 1622:1645 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 1 33 from loss 143:146 arg1 Drosophila 170:179 Drosophila 170:179 Drosophila 170:179 Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. 21949356 8 34 theme EGF 1221:1223 arg1 repeat 1225:1230 the EGF repeat 1217:1230 the EGF repeat 1217:1230 Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. 21949356 3 35 theme identity 415:422 arg1 degree 405:410 a high degree 398:410 a high degree of identity to Drosophila Rumi (52%) 398:447 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 5 36 theme human 714:718 arg1 Rumi 720:723 human Rumi 714:723 human Rumi 714:723 Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. 21949356 7 37 theme repeat 1122:1127 arg1 sequence 1091:1098 the O-glucose consensus sequence 1067:1098 the O-glucose consensus sequence ( ) of factor VII EGF repeat 1067:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 3 38 with others 458:463 arg1 degrees 476:482 lower degrees 470:482 lower degrees of identity 470:494 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 4 39 from UDP-glucose 651:661 arg1 transfer 626:633 transfer 626:633 transfer of glucose from UDP-glucose to an EGF repeat 626:678 Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDP-glucose to an EGF repeat from human factor VII. 21949356 11 40 theme amino 1598:1602 arg1 sequences 1609:1617 amino acid sequences 1598:1617 amino acid sequences of the recipient EGF repeat 1598:1645 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 3 41 theme CAP10 512:516 arg1 domain 518:523 a CAP10 domain 510:523 a CAP10 domain (KDELC1 and KDELC2) 510:543 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 8 42 theme second 1148:1153 arg1 S53 1163:1165 S53 1163:1165 S53 1163:1165 Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. 21949356 8 42 theme second 1148:1153 arg1 serine 1155:1160 the second serine 1144:1160 the second serine (S53) 1144:1166 Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. 21949356 7 43 dep Xyl 1036:1038 arg1 to 1051:1052 to 1051:1052 to 1051:1052 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 7 43 dep Xyl 1036:1038 arg1 serine 1054:1059 serine 52 1054:1062 Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat 1036:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 4 44 theme human 685:689 arg1 VII 698:700 human factor VII 685:700 human factor VII 685:700 Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDP-glucose to an EGF repeat from human factor VII. 21949356 5 45 theme Drosophila 784:793 arg1 clones 800:805 Drosophila rumi clones 784:805 Drosophila rumi clones 784:805 Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. 21949356 8 46 theme Xyl 1192:1194 arg1 transfer 1180:1187 transfer 1180:1187 transfer of Xyl, but not glucose, to the EGF repeat 1180:1230 Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. 21949356 0 47 theme protein 25:31 arg1 Rumi 0:3 Rumi 0:3 Rumi 0:3 Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. 21949356 0 47 theme protein 25:31 arg1 O-glucosyltransferase 33:53 a protein O-glucosyltransferase 23:53 a protein O-glucosyltransferase 23:53 Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. 21949356 2 48 with protein 243:249 arg1 domain 264:269 a CAP10 domain 256:269 a CAP10 domain that functions as a protein O-glucosyltransferase 256:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 48 with protein 243:249 arg1 O-glucosyltransferase 299:319 a protein O-glucosyltransferase 289:319 a protein O-glucosyltransferase 289:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 49 theme Drosophila 182:191 arg1 protein 243:249 a soluble, endoplasmic reticulum-retained protein 201:249 a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase 201:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 49 theme Drosophila 182:191 arg1 Rumi 193:196 Drosophila Rumi 182:196 Drosophila Rumi 182:196 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 9 50 theme consensus 1298:1306 arg1 sequence 1308:1315 the consensus sequence 1294:1315 the consensus sequence 1294:1315 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 10 51 theme O-Xyl 1443:1447 arg1 loss 1435:1438 a loss 1433:1438 a loss of O-Xyl but not O-glucose 1433:1465 Mutation of the second serine (S590A) causes a loss of O-Xyl but not O-glucose at this site. 21949356 6 52 theme Rumi 839:842 arg1 enzymes 844:850 the Rumi enzymes 835:850 the Rumi enzymes 835:850 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 11 53 theme dual 1512:1515 arg1 specificity 1527:1537 dual substrate specificity 1512:1537 dual substrate specificity for the glycosyltransferase Rumi 1512:1570 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 3 54 dep human 325:329 arg1 genomes 341:347 genomes 341:347 genomes 341:347 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 8 55 theme glucose 1205:1211 arg1 transfer 1180:1187 transfer 1180:1187 transfer of Xyl, but not glucose, to the EGF repeat 1180:1230 Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. 21949356 11 56 theme donor 1677:1681 arg1 substrate 1683:1691 donor substrate 1677:1691 donor substrate (UDP-glucose or UDP-Xyl) 1677:1716 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 2 57 theme reticulum-retained 224:241 arg1 protein 243:249 a soluble, endoplasmic reticulum-retained protein 201:249 a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase 201:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 57 theme reticulum-retained 224:241 arg1 Rumi 193:196 Drosophila Rumi 182:196 Drosophila Rumi 182:196 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 5 58 theme rumi 795:798 arg1 clones 800:805 Drosophila rumi clones 784:805 Drosophila rumi clones 784:805 Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. 21949356 3 59 theme high 400:403 arg1 degree 405:410 a high degree 398:410 a high degree of identity to Drosophila Rumi (52%) 398:447 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 11 60 theme acid 1604:1607 arg1 sequences 1609:1617 amino acid sequences 1598:1617 amino acid sequences of the recipient EGF repeat 1598:1645 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 7 61 theme VII 1114:1116 arg1 repeat 1122:1127 factor VII EGF repeat 1107:1127 factor VII EGF repeat 1107:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 7 62 from Xyl 1036:1038 arg1 sequence 1091:1098 the O-glucose consensus sequence 1067:1098 the O-glucose consensus sequence ( ) of factor VII EGF repeat 1067:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 2 63 theme soluble 203:209 arg1 protein 243:249 a soluble, endoplasmic reticulum-retained protein 201:249 a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase 201:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 63 theme soluble 203:209 arg1 Rumi 193:196 Drosophila Rumi 182:196 Drosophila Rumi 182:196 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 1 64 theme temperature-sensitive 121:141 arg1 loss 143:146 a temperature-sensitive loss 119:146 a temperature-sensitive loss of Notch signaling in Drosophila 119:179 Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. 21949356 11 65 theme glycosyltransferase 1547:1565 arg1 Rumi 1567:1570 the glycosyltransferase Rumi 1543:1570 the glycosyltransferase Rumi 1543:1570 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 6 66 dep mammalian 928:936 arg1 Rumi 953:956 Rumi 953:956 Rumi 953:956 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 4 67 theme factor 691:696 arg1 VII 698:700 human factor VII 685:700 human factor VII 685:700 Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDP-glucose to an EGF repeat from human factor VII. 21949356 3 68 dep domain 518:523 arg1 KDELC2 537:542 KDELC2 537:542 KDELC2 537:542 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 3 68 dep domain 518:523 arg1 KDELC1 526:531 KDELC1 526:531 KDELC1 526:531 In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). 21949356 1 69 from Mutations 91:99 arg1 rumi 104:107 rumi 104:107 rumi 104:107 Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. 21949356 4 70 theme glucose 638:644 arg1 transfer 626:633 transfer 626:633 transfer of glucose from UDP-glucose to an EGF repeat 626:678 Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDP-glucose to an EGF repeat from human factor VII. 21949356 0 71 theme protein 61:67 arg1 O-xylosyltransferase 69:88 a protein O-xylosyltransferase 59:88 a protein O-xylosyltransferase 59:88 Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. 21949356 0 71 theme protein 61:67 arg1 Rumi 0:3 Rumi 0:3 Rumi 0:3 Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. 21949356 6 72 theme O-xylosyltransferase 990:1009 arg1 activity 1011:1018 significant protein O-xylosyltransferase activity 970:1018 significant protein O-xylosyltransferase activity 970:1018 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 2 73 theme CAP10 258:262 arg1 domain 264:269 a CAP10 domain 256:269 a CAP10 domain that functions as a protein O-glucosyltransferase 256:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 2 73 theme CAP10 258:262 arg1 O-glucosyltransferase 299:319 a protein O-glucosyltransferase 289:319 a protein O-glucosyltransferase 289:319 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 21949356 9 74 from motif 1285:1289 arg1 sequence 1308:1315 the consensus sequence 1294:1315 the consensus sequence 1294:1315 EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( ), is also modified with either O-Xyl or O-glucose glycans in cells. 21949356 5 75 theme Notch 764:768 arg1 phenotypes 770:779 the Notch phenotypes 760:779 the Notch phenotypes in Drosophila rumi clones 760:805 Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. 21949356 6 76 theme protein 883:889 arg1 activity 913:920 protein O-glucosyltransferase activity 883:920 protein O-glucosyltransferase activity 883:920 During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. 21949356 7 77 theme EGF 1118:1120 arg1 repeat 1122:1127 factor VII EGF repeat 1107:1127 factor VII EGF repeat 1107:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 11 78 theme substrate 1517:1525 arg1 specificity 1527:1537 dual substrate specificity 1512:1537 dual substrate specificity for the glycosyltransferase Rumi 1512:1570 Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used. 21949356 1 79 theme Notch 151:155 arg1 signaling 157:165 Notch signaling 151:165 Notch signaling 151:165 Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. 21949356 7 80 theme O-glucose 1071:1079 arg1 sequence 1091:1098 the O-glucose consensus sequence 1067:1098 the O-glucose consensus sequence ( ) of factor VII EGF repeat 1067:1127 Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( ) of factor VII EGF repeat. 21949356 2 81 dep soluble 203:209 arg1 endoplasmic 212:222 endoplasmic 212:222 endoplasmic 212:222 Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. 23297416 4 0 theme unusual 561:567 arg1 features 580:587 unusual structural features 561:587 unusual structural features that might correlate with this functional diversity 561:639 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 5 1 theme resolution 649:658 arg1 structures 675:684 resolution 1.7- and 1.4-Å structures 649:684 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice 642:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 5 2 from MAdCAM 709:714 arg1 lattice 750:756 a previously described crystal lattice 719:756 a previously described crystal lattice 719:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 7 3 theme I1-set 1067:1072 arg1 fold 1074:1077 The I1-set fold 1063:1077 The I1-set fold 1063:1077 The I1-set fold and CD loop appear biologically relevant. 23297416 5 4 from domains 698:704 arg1 lattice 750:756 a previously described crystal lattice 719:756 a previously described crystal lattice 719:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 8 5 theme different 1125:1133 arg1 conformations 1135:1147 The different conformations 1121:1147 The different conformations seen in crystal structures 1121:1174 The different conformations seen in crystal structures suggest that the integrin-binding loop of MAdCAM is inherently flexible. 23297416 6 6 theme crystal 878:884 arg1 forms 886:890 New crystal forms 874:890 New crystal forms in the presence of two different Fabs to MAdCAM 874:938 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 8 7 theme MAdCAM 1218:1223 arg1 flexible 1239:1246 flexible 1239:1246 flexible 1239:1246 The different conformations seen in crystal structures suggest that the integrin-binding loop of MAdCAM is inherently flexible. 23297416 8 7 theme MAdCAM 1218:1223 arg1 loop 1210:1213 the integrin-binding loop 1189:1213 the integrin-binding loop of MAdCAM 1189:1223 The different conformations seen in crystal structures suggest that the integrin-binding loop of MAdCAM is inherently flexible. 23297416 1 8 theme integrin 166:173 arg1 α4β7 175:178 integrin α4β7 166:178 integrin α4β7 166:178 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 8 9 theme crystal 1157:1163 arg1 structures 1165:1174 crystal structures 1157:1174 crystal structures 1157:1174 The different conformations seen in crystal structures suggest that the integrin-binding loop of MAdCAM is inherently flexible. 23297416 4 10 theme functional 620:629 arg1 diversity 631:639 this functional diversity 615:639 this functional diversity 615:639 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 5 11 theme MAdCAM 709:714 arg1 domains 698:704 the IgSF domains 689:704 the IgSF domains of MAdCAM in a previously described crystal lattice 689:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 5 11 theme MAdCAM 709:714 arg1 MAdCAM 709:714 MAdCAM 709:714 MAdCAM in a previously described crystal lattice 709:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 5 12 theme large 850:854 arg1 contacts 864:871 large lattice contacts 850:871 large lattice contacts 850:871 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 3 13 theme superfamily 311:321 arg1 domains 330:336 the two immunoglobulin superfamily (IgSF) domains 288:336 the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 288:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 13 theme superfamily 311:321 arg1 MAdCAM 341:346 MAdCAM 341:346 MAdCAM 341:346 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 13 theme superfamily 311:321 arg1 α4β7 361:364 integrin α4β7 352:364 integrin α4β7 352:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 6 14 theme Fabs 925:928 arg1 presence 899:906 the presence 895:906 the presence of two different Fabs to MAdCAM 895:938 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 1 15 theme addressin 118:126 arg1 MAdCAM 152:157 MAdCAM 152:157 MAdCAM 152:157 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 1 15 theme addressin 118:126 arg1 molecule 142:149 Mucosal addressin cell adhesion molecule 110:149 Mucosal addressin cell adhesion molecule (MAdCAM) 110:158 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 1 16 theme Mucosal 110:116 arg1 MAdCAM 152:157 MAdCAM 152:157 MAdCAM 152:157 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 1 16 theme Mucosal 110:116 arg1 molecule 142:149 Mucosal addressin cell adhesion molecule 110:149 Mucosal addressin cell adhesion molecule (MAdCAM) 110:158 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 3 17 from ability 384:390 arg1 unusual 369:375 unusual 369:375 unusual 369:375 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 17 from ability 384:390 arg1 interaction 268:278 The interaction 264:278 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 264:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 2 18 theme mucosa-associated 228:244 arg1 tissues 255:261 mucosa-associated lymphoid tissues 228:261 mucosa-associated lymphoid tissues 228:261 Their interaction directs lymphocyte homing to mucosa-associated lymphoid tissues. 23297416 1 19 theme adhesion 133:140 arg1 MAdCAM 152:157 MAdCAM 152:157 MAdCAM 152:157 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 1 19 theme adhesion 133:140 arg1 molecule 142:149 Mucosal addressin cell adhesion molecule 110:149 Mucosal addressin cell adhesion molecule (MAdCAM) 110:158 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 3 20 theme IgSF 324:327 arg1 domains 330:336 the two immunoglobulin superfamily (IgSF) domains 288:336 the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 288:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 20 theme IgSF 324:327 arg1 MAdCAM 341:346 MAdCAM 341:346 MAdCAM 341:346 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 20 theme IgSF 324:327 arg1 α4β7 361:364 integrin α4β7 352:364 integrin α4β7 352:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 0 21 theme I1-set 60:65 arg1 fold 67:70 an I1-set fold 57:70 an I1-set fold 57:70 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 3 22 theme integrin 352:359 arg1 α4β7 361:364 integrin α4β7 352:364 integrin α4β7 352:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 4 23 theme crystal 500:506 arg1 structures 508:517 four crystal structures 495:517 four crystal structures of the IgSF domains of MAdCAM 495:547 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 6 24 theme domain 968:973 arg1 topology 975:982 IgSF domain topology 963:982 IgSF domain topology 963:982 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 3 25 theme MAdCAM 341:346 arg1 domains 330:336 the two immunoglobulin superfamily (IgSF) domains 288:336 the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 288:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 25 theme MAdCAM 341:346 arg1 MAdCAM 341:346 MAdCAM 341:346 MAdCAM 341:346 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 25 theme MAdCAM 341:346 arg1 α4β7 361:364 integrin α4β7 352:364 integrin α4β7 352:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 0 26 theme flexible 78:85 arg1 loop 104:107 a flexible integrin-binding loop 76:107 a flexible integrin-binding loop 76:107 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 9 27 theme corresponding 1285:1297 arg1 loops 1299:1303 the corresponding loops 1281:1303 the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 1281:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 0 28 theme cell 30:33 arg1 molecule 44:51 mucosal addressin cell adhesion molecule 12:51 mucosal addressin cell adhesion molecule 12:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 3 29 theme rolling 410:416 arg1 adhesion 418:425 rolling adhesion 410:425 rolling adhesion 410:425 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 5 30 theme loop 821:824 arg1 conformations 783:795 two alternative conformations 767:795 two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts 767:871 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 9 31 theme different 1509:1517 arg1 conformations 1533:1545 different α4β7 integrin conformations 1509:1545 different α4β7 integrin conformations 1509:1545 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 6 32 theme integrin-binding 1025:1040 arg1 loop 1042:1045 integrin-binding loop 1025:1045 integrin-binding loop 1025:1045 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 5 33 dep Higher 642:647 arg1 structures 675:684 resolution 1.7- and 1.4-Å structures 649:684 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice 642:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 6 34 from shift 954:958 arg1 topology 975:982 IgSF domain topology 963:982 IgSF domain topology 963:982 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 3 35 theme lymphocytes 447:457 arg1 adhesion 418:425 rolling adhesion 410:425 rolling adhesion 410:425 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 35 theme lymphocytes 447:457 arg1 adhesion 435:442 firm adhesion 430:442 firm adhesion 430:442 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 9 36 theme cell 1317:1320 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 36 theme cell 1317:1320 arg1 ICAM-3 1391:1396 ICAM-3 1391:1396 ICAM-3 1391:1396 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 36 theme cell 1317:1320 arg1 ICAM-5 1403:1408 ICAM-5 1403:1408 ICAM-5 1403:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 36 theme cell 1317:1320 arg1 ICAM-2 1383:1388 ICAM-2 1383:1388 ICAM-2 1383:1388 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 36 theme cell 1317:1320 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 6 37 theme New 874:876 arg1 forms 886:890 New crystal forms 874:890 New crystal forms in the presence of two different Fabs to MAdCAM 874:938 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 9 38 theme integrin 1524:1531 arg1 conformations 1533:1545 different α4β7 integrin conformations 1509:1545 different α4β7 integrin conformations 1509:1545 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 0 39 theme addressin 20:28 arg1 molecule 44:51 mucosal addressin cell adhesion molecule 12:51 mucosal addressin cell adhesion molecule 12:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 0 40 theme adhesion 35:42 arg1 molecule 44:51 mucosal addressin cell adhesion molecule 12:51 mucosal addressin cell adhesion molecule 12:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 9 41 theme intercellular 1341:1353 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 41 theme intercellular 1341:1353 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 4 42 theme IgSF 526:529 arg1 MAdCAM 542:547 MAdCAM 542:547 MAdCAM 542:547 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 4 42 theme IgSF 526:529 arg1 domains 531:537 the IgSF domains 522:537 the IgSF domains of MAdCAM 522:547 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 3 43 theme vascular 462:469 arg1 surfaces 471:478 vascular surfaces 462:478 vascular surfaces 462:478 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 9 44 theme molecule 1364:1371 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 44 theme molecule 1364:1371 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 3 45 from adhesion 418:425 arg1 surfaces 471:478 vascular surfaces 462:478 vascular surfaces 462:478 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 7 46 theme CD 1083:1084 arg1 loop 1086:1089 CD loop 1083:1089 CD loop 1083:1089 The I1-set fold and CD loop appear biologically relevant. 23297416 6 47 theme different 915:923 arg1 Fabs 925:928 two different Fabs 911:928 two different Fabs to MAdCAM 911:938 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 6 48 from forms 886:890 arg1 presence 899:906 the presence 895:906 the presence of two different Fabs to MAdCAM 895:938 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 5 49 theme 1.7- 660:663 arg1 structures 675:684 resolution 1.7- and 1.4-Å structures 649:684 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice 642:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 4 50 theme structural 569:578 arg1 features 580:587 unusual structural features 561:587 unusual structural features that might correlate with this functional diversity 561:639 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 5 51 theme lattice 856:862 arg1 contacts 864:871 large lattice contacts 850:871 large lattice contacts 850:871 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 8 52 theme integrin-binding 1193:1208 arg1 flexible 1239:1246 flexible 1239:1246 flexible 1239:1246 The different conformations seen in crystal structures suggest that the integrin-binding loop of MAdCAM is inherently flexible. 23297416 8 52 theme integrin-binding 1193:1208 arg1 loop 1210:1213 the integrin-binding loop 1189:1213 the integrin-binding loop of MAdCAM 1189:1223 The different conformations seen in crystal structures suggest that the integrin-binding loop of MAdCAM is inherently flexible. 23297416 9 53 from loops 1299:1303 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 53 from loops 1299:1303 arg1 ICAM-3 1391:1396 ICAM-3 1391:1396 ICAM-3 1391:1396 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 53 from loops 1299:1303 arg1 ICAM-5 1403:1408 ICAM-5 1403:1408 ICAM-5 1403:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 53 from loops 1299:1303 arg1 ICAM-2 1383:1388 ICAM-2 1383:1388 ICAM-2 1383:1388 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 53 from loops 1299:1303 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 6 54 theme IgSF 963:966 arg1 topology 975:982 IgSF domain topology 963:982 IgSF domain topology 963:982 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 5 55 theme IgSF 693:696 arg1 domains 698:704 the IgSF domains 689:704 the IgSF domains of MAdCAM in a previously described crystal lattice 689:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 5 55 theme IgSF 693:696 arg1 MAdCAM 709:714 MAdCAM 709:714 MAdCAM in a previously described crystal lattice 709:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 9 56 from rigidity 1269:1276 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 56 from rigidity 1269:1276 arg1 ICAM-3 1391:1396 ICAM-3 1391:1396 ICAM-3 1391:1396 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 56 from rigidity 1269:1276 arg1 ICAM-5 1403:1408 ICAM-5 1403:1408 ICAM-5 1403:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 56 from rigidity 1269:1276 arg1 ICAM-2 1383:1388 ICAM-2 1383:1388 ICAM-2 1383:1388 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 56 from rigidity 1269:1276 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 5 57 theme 1.4-Å 669:673 arg1 structures 675:684 resolution 1.7- and 1.4-Å structures 649:684 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice 642:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 2 58 theme lymphocyte 207:216 arg1 homing 218:223 lymphocyte homing 207:223 lymphocyte homing 207:223 Their interaction directs lymphocyte homing to mucosa-associated lymphoid tissues. 23297416 9 59 theme MAdCAM 1446:1451 arg1 specialization 1428:1441 a specialization 1426:1441 a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations 1426:1545 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 3 60 theme immunoglobulin 296:309 arg1 domains 330:336 the two immunoglobulin superfamily (IgSF) domains 288:336 the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 288:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 60 theme immunoglobulin 296:309 arg1 MAdCAM 341:346 MAdCAM 341:346 MAdCAM 341:346 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 60 theme immunoglobulin 296:309 arg1 α4β7 361:364 integrin α4β7 352:364 integrin α4β7 352:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 5 61 theme crystal 742:748 arg1 lattice 750:756 a previously described crystal lattice 719:756 a previously described crystal lattice 719:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 5 62 from lattice 750:756 arg1 domains 698:704 the IgSF domains 689:704 the IgSF domains of MAdCAM in a previously described crystal lattice 689:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 5 62 from lattice 750:756 arg1 MAdCAM 709:714 MAdCAM 709:714 MAdCAM in a previously described crystal lattice 709:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 2 63 theme lymphoid 246:253 arg1 tissues 255:261 mucosa-associated lymphoid tissues 228:261 mucosa-associated lymphoid tissues 228:261 Their interaction directs lymphocyte homing to mucosa-associated lymphoid tissues. 23297416 1 64 theme cell 128:131 arg1 MAdCAM 152:157 MAdCAM 152:157 MAdCAM 152:157 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 1 64 theme cell 128:131 arg1 molecule 142:149 Mucosal addressin cell adhesion molecule 110:149 Mucosal addressin cell adhesion molecule (MAdCAM) 110:158 Mucosal addressin cell adhesion molecule (MAdCAM) binds integrin α4β7. 23297416 6 65 from I2- 993:995 arg1 shift 954:958 a shift 952:958 a shift in IgSF domain topology from the I2- to I1-set 952:1005 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 9 66 from molecule 1331:1338 arg1 rigidity 1269:1276 rigidity 1269:1276 rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 1269:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 67 theme firm 1481:1484 arg1 adhesion 1486:1493 firm adhesion 1481:1493 firm adhesion 1481:1493 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 3 68 theme α4β7 361:364 arg1 domains 330:336 the two immunoglobulin superfamily (IgSF) domains 288:336 the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 288:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 68 theme α4β7 361:364 arg1 MAdCAM 341:346 MAdCAM 341:346 MAdCAM 341:346 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 68 theme α4β7 361:364 arg1 α4β7 361:364 integrin α4β7 352:364 integrin α4β7 352:364 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 0 69 theme integrin-binding 87:102 arg1 loop 104:107 a flexible integrin-binding loop 76:107 a flexible integrin-binding loop 76:107 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 3 70 from unusual 369:375 arg1 ability 384:390 its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces 380:478 its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces 380:478 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 3 71 theme firm 430:433 arg1 adhesion 435:442 firm adhesion 430:442 firm adhesion 430:442 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 5 72 theme integrin-binding 804:819 arg1 loop 821:824 the integrin-binding loop 800:824 the integrin-binding loop 800:824 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 9 73 theme loops 1299:1303 arg1 rigidity 1269:1276 rigidity 1269:1276 rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 1269:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 6 74 theme loop 1042:1045 arg1 switch 1015:1020 a switch 1013:1020 a switch of integrin-binding loop from CC' to CD 1013:1060 New crystal forms in the presence of two different Fabs to MAdCAM demonstrate a shift in IgSF domain topology from the I2- to I1-set, with a switch of integrin-binding loop from CC' to CD. 23297416 4 75 theme MAdCAM 542:547 arg1 MAdCAM 542:547 MAdCAM 542:547 MAdCAM 542:547 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 4 75 theme MAdCAM 542:547 arg1 domains 531:537 the IgSF domains 522:537 the IgSF domains of MAdCAM 522:547 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 0 76 theme mucosal 12:18 arg1 molecule 44:51 mucosal addressin cell adhesion molecule 12:51 mucosal addressin cell adhesion molecule 12:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 9 77 theme vascular 1308:1315 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 77 theme vascular 1308:1315 arg1 ICAM-3 1391:1396 ICAM-3 1391:1396 ICAM-3 1391:1396 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 77 theme vascular 1308:1315 arg1 ICAM-5 1403:1408 ICAM-5 1403:1408 ICAM-5 1403:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 77 theme vascular 1308:1315 arg1 ICAM-2 1383:1388 ICAM-2 1383:1388 ICAM-2 1383:1388 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 77 theme vascular 1308:1315 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 78 theme α4β7 1519:1522 arg1 conformations 1533:1545 different α4β7 integrin conformations 1509:1545 different α4β7 integrin conformations 1509:1545 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 5 79 theme alternative 771:781 arg1 conformations 783:795 two alternative conformations 767:795 two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts 767:871 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 9 80 theme adhesion 1322:1329 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 80 theme adhesion 1322:1329 arg1 ICAM-3 1391:1396 ICAM-3 1391:1396 ICAM-3 1391:1396 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 80 theme adhesion 1322:1329 arg1 ICAM-5 1403:1408 ICAM-5 1403:1408 ICAM-5 1403:1408 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 80 theme adhesion 1322:1329 arg1 ICAM-2 1383:1388 ICAM-2 1383:1388 ICAM-2 1383:1388 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 80 theme adhesion 1322:1329 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 3 81 from adhesion 435:442 arg1 surfaces 471:478 vascular surfaces 462:478 vascular surfaces 462:478 The interaction between the two immunoglobulin superfamily (IgSF) domains of MAdCAM and integrin α4β7 is unusual in its ability to mediate either rolling adhesion or firm adhesion of lymphocytes on vascular surfaces. 23297416 5 82 theme described 732:740 arg1 lattice 750:756 a previously described crystal lattice 719:756 a previously described crystal lattice 719:756 Higher resolution 1.7- and 1.4-Å structures of the IgSF domains of MAdCAM in a previously described crystal lattice revealed two alternative conformations of the integrin-binding loop, which were deformed by large lattice contacts. 23297416 0 83 theme molecule 44:51 arg1 Domain 0:5 Domain 1 0:7 Domain 1 of mucosal addressin cell adhesion molecule 0:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 0 83 theme molecule 44:51 arg1 molecule 44:51 mucosal addressin cell adhesion molecule 12:51 mucosal addressin cell adhesion molecule 12:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 4 84 theme domains 531:537 arg1 structures 508:517 four crystal structures 495:517 four crystal structures of the IgSF domains of MAdCAM 495:547 We determined four crystal structures of the IgSF domains of MAdCAM to test for unusual structural features that might correlate with this functional diversity. 23297416 9 85 theme adhesion 1355:1362 arg1 ICAM 1374:1377 intercellular adhesion molecule (ICAM)-1 1341:1380 intercellular adhesion molecule (ICAM)-1 1341:1380 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 9 85 theme adhesion 1355:1362 arg1 molecule 1331:1338 vascular cell adhesion molecule 1308:1338 vascular cell adhesion molecule 1308:1338 This contrasts with rigidity of the corresponding loops in vascular cell adhesion molecule, intercellular adhesion molecule (ICAM)-1, ICAM-2, ICAM-3, and ICAM-5 and may reflect a specialization of MAdCAM to mediate both rolling and firm adhesion by binding to different α4β7 integrin conformations. 23297416 0 86 contain has 53:55 arg2 loop 104:107 a flexible integrin-binding loop 76:107 a flexible integrin-binding loop 76:107 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 0 86 contain has 53:55 arg1 Domain 0:5 Domain 1 0:7 Domain 1 of mucosal addressin cell adhesion molecule 0:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 0 86 contain has 53:55 arg2 fold 67:70 an I1-set fold 57:70 an I1-set fold 57:70 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 23297416 0 86 contain has 53:55 arg1 molecule 44:51 mucosal addressin cell adhesion molecule 12:51 mucosal addressin cell adhesion molecule 12:51 Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. 17395589 4 0 mod modified 549:556 arg1 TSRs 540:543 the TSRs 536:543 the TSRs 536:543 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 4 0 mod modified 549:556 arg1 six 529:531 six 529:531 six 529:531 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 4 0 mod modified 549:556 arg3 disaccharide 575:586 an O-fucose disaccharide 563:586 an O-fucose disaccharide 563:586 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 3 1 contain contain 326:332 arg1 repeats 296:302 eight thrombospondin type 1 repeats 268:302 eight thrombospondin type 1 repeats (TSR) 268:308 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 1 contain contain 326:332 arg2 sequence 346:353 a consensus sequence 334:353 a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine 334:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 1 contain contain 326:332 arg1 TSR 305:307 TSR 305:307 TSR 305:307 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 1 contain contain 326:332 arg1 seven 311:315 seven 311:315 seven 311:315 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 9 2 theme fucose 1223:1228 arg1 addition 1230:1237 fucose addition 1223:1237 fucose addition 1223:1237 A similar outcome was observed when ADAMTS13 was expressed in a cell line unable to synthesize the donor for fucose addition, GDP-fucose. 17395589 9 2 theme fucose 1223:1228 arg1 GDP-fucose 1240:1249 GDP-fucose 1240:1249 GDP-fucose 1240:1249 A similar outcome was observed when ADAMTS13 was expressed in a cell line unable to synthesize the donor for fucose addition, GDP-fucose. 17395589 10 3 theme TSR1,2 1384:1389 arg1 mutant 1398:1403 the ADAMTS13 TSR1,2 double mutant 1371:1403 the ADAMTS13 TSR1,2 double mutant 1371:1403 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 10 4 theme wild-type 1318:1326 arg1 ADAMTS13 1328:1335 wild-type ADAMTS13 1318:1335 wild-type ADAMTS13 1318:1335 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 4 5 attach derived 478:484 arg1 ADAMTS13 497:504 human ADAMTS13 491:504 human ADAMTS13 491:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 4 5 attach derived 478:484 arg2 peptides 469:476 tryptic peptides 461:476 tryptic peptides derived from human ADAMTS13 461:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 4 6 theme Mass 435:438 arg1 analysis 449:456 Mass spectral analysis 435:456 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 435:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 5 7 theme fucose 607:612 arg1 Analysis 589:596 Analysis 589:596 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 589:653 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 demonstrated that the disaccharide has the structure glucose-beta1,3-fucose. 17395589 1 8 dep smaller 128:134 arg1 thrombogenic 142:153 thrombogenic 142:153 thrombogenic 142:153 ADAMTS13 is a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms. 17395589 10 9 theme POFUT2 1279:1284 arg1 overexpression 1261:1274 overexpression 1261:1274 overexpression of POFUT2 1261:1284 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 4 10 theme O-fucose 566:573 arg1 disaccharide 575:586 an O-fucose disaccharide 563:586 an O-fucose disaccharide 563:586 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 4 11 theme tryptic 461:467 arg1 peptides 469:476 tryptic peptides 461:476 tryptic peptides derived from human ADAMTS13 461:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 9 12 theme similar 1116:1122 arg1 outcome 1124:1130 A similar outcome 1114:1130 A similar outcome 1114:1130 A similar outcome was observed when ADAMTS13 was expressed in a cell line unable to synthesize the donor for fucose addition, GDP-fucose. 17395589 11 13 theme ADAMTS13 1543:1550 arg1 secretion 1530:1538 secretion 1530:1538 secretion of ADAMTS13 1530:1550 Together these findings indicate that O-fucosylation is functionally significant for secretion of ADAMTS13. 17395589 3 14 theme consensus 336:344 arg1 sequence 346:353 a consensus sequence 334:353 a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine 334:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 10 15 theme double 1391:1396 arg1 mutant 1398:1403 the ADAMTS13 TSR1,2 double mutant 1371:1403 the ADAMTS13 TSR1,2 double mutant 1371:1403 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 1 16 theme plasma 65:70 arg1 metalloproteinase 72:88 a plasma metalloproteinase 63:88 a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms 63:159 ADAMTS13 is a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms. 17395589 1 16 theme plasma 65:70 arg1 ADAMTS13 51:58 ADAMTS13 51:58 ADAMTS13 51:58 ADAMTS13 is a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms. 17395589 8 17 theme ADAMTS13 1094:1101 arg1 secretion 1081:1089 the secretion 1077:1089 the secretion of ADAMTS13 1077:1101 When the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, was reduced using siRNA, the secretion of ADAMTS13 decreased. 17395589 3 18 theme thrombospondin 274:287 arg1 type 289:292 thrombospondin type 1 274:294 eight thrombospondin type 1 repeats (TSR) 268:308 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 8 19 theme protein 958:964 arg1 enzyme 1003:1008 the enzyme 999:1008 the enzyme that transfers fucose to serines in TSRs 999:1049 When the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, was reduced using siRNA, the secretion of ADAMTS13 decreased. 17395589 8 19 theme protein 958:964 arg1 O-fucosyltransferase 966:985 protein O-fucosyltransferase 2 958:987 protein O-fucosyltransferase 2 (POFUT2) 958:996 When the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, was reduced using siRNA, the secretion of ADAMTS13 decreased. 17395589 8 19 theme protein 958:964 arg1 POFUT2 990:995 POFUT2 990:995 POFUT2 990:995 When the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, was reduced using siRNA, the secretion of ADAMTS13 decreased. 17395589 3 20 theme threonine 424:432 arg1 threonine 424:432 threonine 424:432 threonine 424:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 20 theme threonine 424:432 arg1 serine 414:419 serine 414:419 serine 414:419 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 20 theme threonine 424:432 arg1 group 405:409 the hydroxyl group 392:409 the hydroxyl group of serine or threonine 392:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 5 21 dep fucose 607:612 arg1 incorporated 628:639 incorporated 628:639 metabolically incorporated into ADAMTS13 614:653 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 demonstrated that the disaccharide has the structure glucose-beta1,3-fucose. 17395589 4 22 theme peptides 469:476 arg1 analysis 449:456 Mass spectral analysis 435:456 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 435:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 10 23 theme ADAMTS13 1375:1382 arg1 mutant 1398:1403 the ADAMTS13 TSR1,2 double mutant 1371:1403 the ADAMTS13 TSR1,2 double mutant 1371:1403 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 3 24 theme fucose 382:387 arg1 addition 370:377 the direct addition 359:377 the direct addition of fucose to the hydroxyl group of serine or threonine 359:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 5 25 contain has 690:692 arg1 disaccharide 677:688 the disaccharide 673:688 the disaccharide 673:688 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 demonstrated that the disaccharide has the structure glucose-beta1,3-fucose. 17395589 5 25 contain has 690:692 arg2 structure 698:706 the structure 694:706 the structure glucose-beta1,3-fucose 694:729 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 demonstrated that the disaccharide has the structure glucose-beta1,3-fucose. 17395589 3 26 theme serine 414:419 arg1 threonine 424:432 threonine 424:432 threonine 424:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 26 theme serine 414:419 arg1 serine 414:419 serine 414:419 serine 414:419 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 26 theme serine 414:419 arg1 group 405:409 the hydroxyl group 392:409 the hydroxyl group of serine or threonine 392:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 27 theme direct 363:368 arg1 addition 370:377 the direct addition 359:377 the direct addition of fucose to the hydroxyl group of serine or threonine 359:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 28 theme repeats 296:302 arg1 repeats 296:302 eight thrombospondin type 1 repeats 268:302 eight thrombospondin type 1 repeats (TSR) 268:308 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 28 theme repeats 296:302 arg1 TSR 305:307 TSR 305:307 TSR 305:307 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 28 theme repeats 296:302 arg1 seven 311:315 seven 311:315 seven 311:315 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 6 29 theme serine 757:762 arg1 Mutation 732:739 Mutation 732:739 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 732:803 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 1 30 theme von 103:105 arg1 factor 118:123 von Willebrand factor 103:123 von Willebrand factor to smaller, less thrombogenic forms 103:159 ADAMTS13 is a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms. 17395589 10 31 theme ADAMTS13 1328:1335 arg1 secretion 1305:1313 the secretion 1301:1313 the secretion of wild-type ADAMTS13 1301:1335 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 2 32 theme activity 185:192 arg1 Deficiency 162:171 Deficiency 162:171 Deficiency of ADAMTS13 activity in plasma 162:202 Deficiency of ADAMTS13 activity in plasma leads to thrombotic thrombocytopenic purpura. 17395589 1 33 theme Willebrand 107:116 arg1 factor 118:123 von Willebrand factor 103:123 von Willebrand factor to smaller, less thrombogenic forms 103:159 ADAMTS13 is a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms. 17395589 3 34 theme hydroxyl 396:403 arg1 threonine 424:432 threonine 424:432 threonine 424:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 34 theme hydroxyl 396:403 arg1 serine 414:419 serine 414:419 serine 414:419 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 34 theme hydroxyl 396:403 arg1 group 405:409 the hydroxyl group 392:409 the hydroxyl group of serine or threonine 392:432 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 4 35 theme TSRs 540:543 arg1 six 529:531 six 529:531 six 529:531 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 4 35 theme TSRs 540:543 arg1 TSRs 540:543 the TSRs 536:543 the TSRs 536:543 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 3 36 theme type 289:292 arg1 repeats 296:302 eight thrombospondin type 1 repeats 268:302 eight thrombospondin type 1 repeats (TSR) 268:308 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 36 theme type 289:292 arg1 TSR 305:307 TSR 305:307 TSR 305:307 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 2 37 theme ADAMTS13 176:183 arg1 activity 185:192 ADAMTS13 activity 176:192 ADAMTS13 activity 176:192 Deficiency of ADAMTS13 activity in plasma leads to thrombotic thrombocytopenic purpura. 17395589 0 38 gly O-fucosylation 0:13 arg1 ADAMTS13 31:38 ADAMTS13 secretion 31:48 ADAMTS13 secretion 31:48 O-fucosylation is required for ADAMTS13 secretion. 17395589 6 39 from Mutation 732:739 arg1 TSR7 790:793 TSR7 790:793 TSR7 790:793 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 6 39 from Mutation 732:739 arg1 TSR8 800:803 TSR8 800:803 TSR8 800:803 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 6 39 from Mutation 732:739 arg1 TSR5 784:787 TSR5 784:787 TSR5 784:787 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 6 39 from Mutation 732:739 arg1 TSR2 778:781 TSR2 778:781 TSR2 778:781 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 0 40 theme ADAMTS13 31:38 arg1 secretion 40:48 ADAMTS13 secretion 31:48 ADAMTS13 secretion 31:48 O-fucosylation is required for ADAMTS13 secretion. 17395589 10 41 theme mutant 1398:1403 arg1 secretion 1358:1366 the secretion 1354:1366 the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant 1354:1442 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 5 42 dep H 605:605 arg1 3 603:603 3 603:603 3 603:603 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 demonstrated that the disaccharide has the structure glucose-beta1,3-fucose. 17395589 6 43 theme modified 748:755 arg1 serine 757:762 the modified serine 744:762 the modified serine 744:762 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 4 44 theme spectral 440:447 arg1 analysis 449:456 Mass spectral analysis 435:456 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 435:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 17395589 2 45 theme thrombocytopenic 224:239 arg1 purpura 241:247 thrombotic thrombocytopenic purpura 213:247 thrombotic thrombocytopenic purpura 213:247 Deficiency of ADAMTS13 activity in plasma leads to thrombotic thrombocytopenic purpura. 17395589 10 46 theme TSR1-8 1430:1435 arg1 mutant 1437:1442 ADAMTS13 TSR1-8 mutant 1421:1442 ADAMTS13 TSR1-8 mutant 1421:1442 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 9 47 theme cell 1178:1181 arg1 line 1183:1186 a cell line 1176:1186 a cell line unable to synthesize the donor for fucose addition, GDP-fucose 1176:1249 A similar outcome was observed when ADAMTS13 was expressed in a cell line unable to synthesize the donor for fucose addition, GDP-fucose. 17395589 2 48 theme thrombotic 213:222 arg1 purpura 241:247 thrombotic thrombocytopenic purpura 213:247 thrombotic thrombocytopenic purpura 213:247 Deficiency of ADAMTS13 activity in plasma leads to thrombotic thrombocytopenic purpura. 17395589 6 49 theme ADAMTS13 830:837 arg1 secretion 817:825 the secretion 813:825 the secretion of ADAMTS13 813:837 Mutation of the modified serine to alanine in TSR2, TSR5, TSR7, and TSR8 reduced the secretion of ADAMTS13. 17395589 3 50 contain contains 259:266 arg2 TSR 305:307 TSR 305:307 TSR 305:307 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 50 contain contains 259:266 arg1 ADAMTS13 250:257 ADAMTS13 250:257 ADAMTS13 250:257 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 3 50 contain contains 259:266 arg2 repeats 296:302 eight thrombospondin type 1 repeats 268:302 eight thrombospondin type 1 repeats (TSR) 268:308 ADAMTS13 contains eight thrombospondin type 1 repeats (TSR), seven of which contain a consensus sequence for the direct addition of fucose to the hydroxyl group of serine or threonine. 17395589 8 51 theme O-fucosyltransferase 966:985 arg1 expression 944:953 the expression 940:953 the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, 940:1050 When the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, was reduced using siRNA, the secretion of ADAMTS13 decreased. 17395589 10 52 theme ADAMTS13 1421:1428 arg1 mutant 1437:1442 ADAMTS13 TSR1-8 mutant 1421:1442 ADAMTS13 TSR1-8 mutant 1421:1442 Although overexpression of POFUT2 did not affect the secretion of wild-type ADAMTS13, it did increase the secretion of the ADAMTS13 TSR1,2 double mutant but not that of ADAMTS13 TSR1-8 mutant. 17395589 1 53 theme smaller 128:134 arg1 forms 155:159 smaller, less thrombogenic forms 128:159 smaller, less thrombogenic forms 128:159 ADAMTS13 is a plasma metalloproteinase that cleaves von Willebrand factor to smaller, less thrombogenic forms. 17395589 7 54 theme site 866:869 arg1 Mutation 840:847 Mutation 840:847 Mutation of more than one site 840:869 Mutation of more than one site dramatically reduced secretion regardless of the sites mutated. 17395589 5 55 theme glucose-beta1,3-fucose 708:729 arg1 structure 698:706 the structure 694:706 the structure glucose-beta1,3-fucose 694:729 Analysis of [(3)H]fucose metabolically incorporated into ADAMTS13 demonstrated that the disaccharide has the structure glucose-beta1,3-fucose. 17395589 9 56 theme unable 1188:1193 arg1 line 1183:1186 a cell line 1176:1186 a cell line unable to synthesize the donor for fucose addition, GDP-fucose 1176:1249 A similar outcome was observed when ADAMTS13 was expressed in a cell line unable to synthesize the donor for fucose addition, GDP-fucose. 17395589 8 57 from serines 1035:1041 arg1 TSRs 1046:1049 TSRs 1046:1049 TSRs 1046:1049 When the expression of protein O-fucosyltransferase 2 (POFUT2), the enzyme that transfers fucose to serines in TSRs, was reduced using siRNA, the secretion of ADAMTS13 decreased. 17395589 2 58 from Deficiency 162:171 arg1 plasma 197:202 plasma 197:202 plasma 197:202 Deficiency of ADAMTS13 activity in plasma leads to thrombotic thrombocytopenic purpura. 17395589 4 59 theme human 491:495 arg1 ADAMTS13 497:504 human ADAMTS13 491:504 human ADAMTS13 491:504 Mass spectral analysis of tryptic peptides derived from human ADAMTS13 indicate that at least six of the TSRs are modified with an O-fucose disaccharide. 10899108 4 0 from p40 570:572 arg1 residues 548:555 Several charged residues 532:555 Several charged residues from p35 and p40 532:572 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 1 1 theme early 145:149 arg1 cytokine 168:175 an early pro-inflammatory cytokine 142:175 an early pro-inflammatory cytokine 142:175 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 1 theme early 145:149 arg1 interleukin-12 111:124 Human interleukin-12 105:124 Human interleukin-12 (IL-12, p70) 105:137 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 0 2 theme cytokine 80:87 arg1 interleukin-12 89:102 the heterodimeric cytokine interleukin-12 62:102 the heterodimeric cytokine interleukin-12 62:102 Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. 10899108 5 3 theme IL-12 819:823 arg1 formation 825:833 IL-12 formation 819:833 IL-12 formation 819:833 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 1 4 theme pro-inflammatory 151:166 arg1 cytokine 168:175 an early pro-inflammatory cytokine 142:175 an early pro-inflammatory cytokine 142:175 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 4 theme pro-inflammatory 151:166 arg1 interleukin-12 111:124 Human interleukin-12 105:124 Human interleukin-12 (IL-12, p70) 105:137 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 4 5 theme charged 540:546 arg1 residues 548:555 Several charged residues 532:555 Several charged residues from p35 and p40 532:572 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 4 6 from p35 562:564 arg1 residues 548:555 Several charged residues 532:555 Several charged residues from p35 and p40 532:572 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 1 7 link disulfide-linked 193:208 arg1 subunits 210:217 two disulfide-linked subunits 189:217 two disulfide-linked subunits 189:217 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 7 link disulfide-linked 193:208 arg1 p40 228:230 p40 228:230 p40 228:230 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 7 link disulfide-linked 193:208 arg1 p35 220:222 p35 220:222 p35 220:222 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 2 8 theme p40 285:287 arg1 complex 316:322 the human p70 complex 302:322 the human p70 complex 302:322 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 2 8 theme p40 285:287 arg1 structures 255:264 the crystal structures 243:264 the crystal structures of monomeric human p40 at 2.5 A 243:296 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 2 9 from A 296:296 arg1 complex 316:322 the human p70 complex 302:322 the human p70 complex 302:322 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 2 9 from A 296:296 arg1 structures 255:264 the crystal structures 243:264 the crystal structures of monomeric human p40 at 2.5 A 243:296 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 5 10 theme central 685:691 arg1 residue 702:708 A central arginine residue 683:708 A central arginine residue from p35 683:717 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 2 11 theme human 279:283 arg1 p40 285:287 monomeric human p40 269:287 monomeric human p40 269:287 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 5 12 theme arginine 693:700 arg1 residue 702:708 A central arginine residue 683:708 A central arginine residue from p35 683:717 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 0 13 from topography 48:57 arg1 interleukin-12 89:102 the heterodimeric cytokine interleukin-12 62:102 the heterodimeric cytokine interleukin-12 62:102 Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. 10899108 2 14 theme monomeric 269:277 arg1 p40 285:287 monomeric human p40 269:287 monomeric human p40 269:287 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 5 15 theme formation 825:833 arg1 antagonist 805:814 a small molecule antagonist 788:814 a small molecule antagonist of IL-12 formation 788:833 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 0 16 theme Charged 0:6 arg1 residues 8:15 Charged residues 0:15 Charged residues 0:15 Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. 10899108 5 17 theme small 790:794 arg1 antagonist 805:814 a small molecule antagonist 788:814 a small molecule antagonist of IL-12 formation 788:833 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 2 18 theme cytokine-receptor 392:408 arg1 complexes 410:418 class 1 cytokine-receptor complexes 384:418 class 1 cytokine-receptor complexes 384:418 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 1 19 theme Human 105:109 arg1 cytokine 168:175 an early pro-inflammatory cytokine 142:175 an early pro-inflammatory cytokine 142:175 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 19 theme Human 105:109 arg1 interleukin-12 111:124 Human interleukin-12 105:124 Human interleukin-12 (IL-12, p70) 105:137 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 5 20 theme molecule 796:803 arg1 antagonist 805:814 a small molecule antagonist 788:814 a small molecule antagonist of IL-12 formation 788:833 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 3 21 theme p40 519:521 arg1 subunit 523:529 the p40 subunit 515:529 the p40 subunit 515:529 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 10899108 1 22 theme disulfide-linked 193:208 arg1 subunits 210:217 two disulfide-linked subunits 189:217 two disulfide-linked subunits 189:217 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 22 theme disulfide-linked 193:208 arg1 p40 228:230 p40 228:230 p40 228:230 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 22 theme disulfide-linked 193:208 arg1 p35 220:222 p35 220:222 p35 220:222 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 2 23 theme class 384:388 arg1 complexes 410:418 class 1 cytokine-receptor complexes 384:418 class 1 cytokine-receptor complexes 384:418 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 4 24 theme Several 532:538 arg1 residues 548:555 Several charged residues 532:555 Several charged residues from p35 and p40 532:572 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 3 25 theme N-terminal 467:476 arg1 domain 498:503 an N-terminal immunoglobulin-like domain 464:503 an N-terminal immunoglobulin-like domain 464:503 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 10899108 3 26 located found 506:510 arg2 domain 498:503 an N-terminal immunoglobulin-like domain 464:503 an N-terminal immunoglobulin-like domain 464:503 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 10899108 3 26 located found 506:510 arg1 subunit 523:529 the p40 subunit 515:529 the p40 subunit 515:529 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 10899108 3 27 theme immunoglobulin-like 478:496 arg1 domain 498:503 an N-terminal immunoglobulin-like domain 464:503 an N-terminal immunoglobulin-like domain 464:503 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 10899108 0 28 theme interlocking 35:46 arg1 topography 48:57 a unique interlocking topography 26:57 a unique interlocking topography in the heterodimeric cytokine interleukin-12 26:102 Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. 10899108 5 29 theme ideal 771:775 arg1 pocket 740:745 a deep pocket 733:745 a deep pocket 733:745 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 5 29 theme ideal 771:775 arg1 target 777:782 an ideal target 768:782 an ideal target for a small molecule antagonist of IL-12 formation 768:833 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 4 30 theme interlocking 603:614 arg1 topography 616:625 a unique interlocking topography 594:625 a unique interlocking topography 594:625 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 0 31 theme unique 28:33 arg1 topography 48:57 a unique interlocking topography 26:57 a unique interlocking topography in the heterodimeric cytokine interleukin-12 26:102 Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. 10899108 5 32 theme deep 735:738 arg1 pocket 740:745 a deep pocket 733:745 a deep pocket 733:745 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 5 32 theme deep 735:738 arg1 target 777:782 an ideal target 768:782 an ideal target for a small molecule antagonist of IL-12 formation 768:833 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 4 33 theme unique 596:601 arg1 topography 616:625 a unique interlocking topography 594:625 a unique interlocking topography 594:625 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 4 34 theme p70 668:670 arg1 formation 672:680 p70 formation 668:680 p70 formation 668:680 Several charged residues from p35 and p40 intercalate to form a unique interlocking topography, shown by mutagenesis to be critical for p70 formation. 10899108 2 35 theme p70 312:314 arg1 complex 316:322 the human p70 complex 302:322 the human p70 complex 302:322 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 2 36 theme human 306:310 arg1 complex 316:322 the human p70 complex 302:322 the human p70 complex 302:322 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 2 37 theme crystal 247:253 arg1 structures 255:264 the crystal structures 243:264 the crystal structures of monomeric human p40 at 2.5 A 243:296 We solved the crystal structures of monomeric human p40 at 2.5 A and the human p70 complex at 2.8 A resolution, which reveals that IL-12 is similar to class 1 cytokine-receptor complexes. 10899108 3 38 theme first 443:447 arg1 description 449:459 the first description 439:459 the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit 439:529 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 10899108 0 39 theme heterodimeric 66:78 arg1 interleukin-12 89:102 the heterodimeric cytokine interleukin-12 62:102 the heterodimeric cytokine interleukin-12 62:102 Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. 10899108 5 40 from p35 715:717 arg1 residue 702:708 A central arginine residue 683:708 A central arginine residue from p35 683:717 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 1 41 dep interleukin-12 111:124 arg1 p70 134:136 p70 134:136 p70 134:136 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 1 41 dep interleukin-12 111:124 arg1 IL-12 127:131 IL-12 127:131 IL-12 127:131 Human interleukin-12 (IL-12, p70) is an early pro-inflammatory cytokine, comprising two disulfide-linked subunits, p35 and p40. 10899108 5 42 from pocket 740:745 arg1 p40 750:752 p40 750:752 p40 750:752 A central arginine residue from p35 projects into a deep pocket on p40, which may be an ideal target for a small molecule antagonist of IL-12 formation. 10899108 3 43 theme domain 498:503 arg1 description 449:459 the first description 439:459 the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit 439:529 They also include the first description of an N-terminal immunoglobulin-like domain, found on the p40 subunit. 30918116 5 0 theme PANX1 652:656 arg1 activity 666:673 aberrant PANX1 channel activity 643:673 aberrant PANX1 channel activity 643:673 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 5 1 from release 680:686 arg1 oocytes 691:697 oocytes 691:697 oocytes 691:697 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 5 2 theme PANX1 533:537 arg1 pattern 553:559 the PANX1 glycosylation pattern 529:559 the PANX1 glycosylation pattern 529:559 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 6 3 theme mutation 759:766 arg1 Overexpression 723:736 Overexpression 723:736 Overexpression of a patient-derived mutation in mice 723:774 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 1 4 theme cellular 134:141 arg1 communication 143:155 cellular communication 134:155 cellular communication 134:155 Connexins and pannexins are two protein families that play an important role in cellular communication. 30918116 4 5 dep describe 314:321 arg1 " 466:466 " 466:466 " 466:466 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 5 6 theme channel 658:664 arg1 activity 666:673 aberrant PANX1 channel activity 643:673 aberrant PANX1 channel activity 643:673 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 5 7 from activity 666:673 arg1 oocytes 691:697 oocytes 691:697 oocytes 691:697 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 5 8 theme glycosylation 539:551 arg1 pattern 553:559 the PANX1 glycosylation pattern 529:559 the PANX1 glycosylation pattern 529:559 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 7 9 theme critical 874:881 arg1 role 883:886 the critical role 870:886 the critical role of PANX1 in human oocyte development 870:923 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 4 10 theme familial 383:390 arg1 infertility 411:421 familial or sporadic female infertility 383:421 familial or sporadic female infertility 383:421 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 7 11 theme infertility 973:983 arg1 subtype 962:968 a subtype 960:968 a subtype of infertility 960:983 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 5 12 theme oocyte 509:514 arg1 death 516:520 oocyte death 509:520 oocyte death 509:520 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 2 13 theme channel 222:228 arg1 protein 230:236 a channel protein 220:236 a channel protein 220:236 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 30918116 2 13 theme channel 222:228 arg1 Pannexin 158:165 Pannexin 1 158:167 Pannexin 1 (PANX1) 158:175 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 30918116 4 14 theme female 404:409 arg1 infertility 411:421 familial or sporadic female infertility 383:421 familial or sporadic female infertility 383:421 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 7 15 theme therapeutic 1021:1031 arg1 intervention 1033:1044 therapeutic intervention 1021:1044 therapeutic intervention for this disease 1021:1061 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 0 16 theme pannexin 2:9 arg1 channelopathy 13:25 A pannexin 1 channelopathy 0:25 A pannexin 1 channelopathy 0:25 A pannexin 1 channelopathy causes human oocyte death. 30918116 4 17 theme sporadic 395:402 arg1 infertility 411:421 familial or sporadic female infertility 383:421 familial or sporadic female infertility 383:421 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 7 18 from role 883:886 arg1 development 913:923 human oocyte development 900:923 human oocyte development 900:923 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 6 19 from Overexpression 723:736 arg1 mice 771:774 mice 771:774 mice 771:774 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 7 20 theme potential 1000:1008 arg1 target 1010:1015 a potential target 998:1015 a potential target for therapeutic intervention for this disease 998:1061 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 4 21 theme oocyte 453:458 arg1 death 460:464 "oocyte death 452:464 "oocyte death 452:464 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 1 22 theme protein 86:92 arg1 pannexins 68:76 pannexins 68:76 pannexins 68:76 Connexins and pannexins are two protein families that play an important role in cellular communication. 30918116 1 22 theme protein 86:92 arg1 Connexins 54:62 Connexins 54:62 Connexins 54:62 Connexins and pannexins are two protein families that play an important role in cellular communication. 30918116 1 22 theme protein 86:92 arg1 families 94:101 two protein families 82:101 two protein families that play an important role in cellular communication 82:155 Connexins and pannexins are two protein families that play an important role in cellular communication. 30918116 5 23 theme mutant 704:709 arg1 GLY1 717:720 mutant PANX1 GLY1 704:720 mutant PANX1 GLY1 704:720 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 5 24 theme PANX1 604:608 arg1 localization 588:599 the subcellular localization 572:599 the subcellular localization of PANX1 in cultured cells 572:626 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 6 25 theme patient-derived 743:757 arg1 mutation 759:766 a patient-derived mutation 741:766 a patient-derived mutation 741:766 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 5 26 theme PANX1 711:715 arg1 GLY1 717:720 mutant PANX1 GLY1 704:720 mutant PANX1 GLY1 704:720 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 0 27 theme human 34:38 arg1 death 47:51 human oocyte death 34:51 human oocyte death 34:51 A pannexin 1 channelopathy causes human oocyte death. 30918116 7 28 theme genetic 936:942 arg1 explanation 944:954 a genetic explanation 934:954 a genetic explanation for a subtype of infertility 934:983 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 5 29 theme cultured 613:620 arg1 cells 622:626 cultured cells 613:626 cultured cells 613:626 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 4 30 from mutations 358:366 arg1 PANX1 371:375 PANX1 371:375 PANX1 371:375 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 2 31 theme members 189:195 arg1 members 189:195 the members 185:195 the members of pannexin family 185:214 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 30918116 2 31 theme members 189:195 arg1 Pannexin 158:165 Pannexin 1 158:167 Pannexin 1 (PANX1) 158:175 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 30918116 2 31 theme members 189:195 arg1 one 178:180 one 178:180 one 178:180 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 30918116 5 32 from GLY1 717:720 arg1 oocytes 691:697 oocytes 691:697 oocytes 691:697 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 5 33 theme ATP 676:678 arg1 release 680:686 ATP release 676:686 ATP release in oocytes 676:697 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 6 34 theme death 828:832 arg1 phenotype 834:842 the human oocyte death phenotype 811:842 the human oocyte death phenotype 811:842 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 5 35 theme aberrant 643:650 arg1 activity 666:673 aberrant PANX1 channel activity 643:673 aberrant PANX1 channel activity 643:673 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 7 36 theme oocyte 906:911 arg1 development 913:923 human oocyte development 900:923 human oocyte development 900:923 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 0 37 theme oocyte 40:45 arg1 death 47:51 human oocyte death 34:51 human oocyte death 34:51 A pannexin 1 channelopathy causes human oocyte death. 30918116 6 38 theme oocyte 821:826 arg1 phenotype 834:842 the human oocyte death phenotype 811:842 the human oocyte death phenotype 811:842 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 7 39 theme PANX1 891:895 arg1 role 883:886 the critical role 870:886 the critical role of PANX1 in human oocyte development 870:923 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 6 40 theme human 815:819 arg1 phenotype 834:842 the human oocyte death phenotype 811:842 the human oocyte death phenotype 811:842 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 5 41 theme subcellular 576:586 arg1 localization 588:599 the subcellular localization 572:599 the subcellular localization of PANX1 in cultured cells 572:626 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 4 42 theme independent 328:338 arg1 families 340:347 four independent families 323:347 four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death 323:464 Here, we describe four independent families in which mutations in PANX1 cause familial or sporadic female infertility via a phenotype that we term "oocyte death." 30918116 6 43 link patient-derived 743:757 arg1 mutation 759:766 a patient-derived mutation 741:766 a patient-derived mutation 741:766 Overexpression of a patient-derived mutation in mice causes infertility, recapitulating the human oocyte death phenotype. 30918116 7 44 theme human 900:904 arg1 development 913:923 human oocyte development 900:923 human oocyte development 900:923 Our findings demonstrate the critical role of PANX1 in human oocyte development, provide a genetic explanation for a subtype of infertility, and suggest a potential target for therapeutic intervention for this disease. 30918116 1 45 theme important 116:124 arg1 role 126:129 an important role 113:129 an important role 113:129 Connexins and pannexins are two protein families that play an important role in cellular communication. 30918116 5 46 from localization 588:599 arg1 cells 622:626 cultured cells 613:626 cultured cells 613:626 The mutations, which are associated with oocyte death, alter the PANX1 glycosylation pattern, influence the subcellular localization of PANX1 in cultured cells, and result in aberrant PANX1 channel activity, ATP release in oocytes, and mutant PANX1 GLY1. 30918116 2 47 theme family 209:214 arg1 members 189:195 the members 185:195 the members of pannexin family 185:214 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 30918116 3 48 gly glycosylated 245:256 arg1 It 239:240 It 239:240 It 239:240 It is glycosylated and forms three species, GLY0, GLY1, and GLY2. 30918116 2 49 theme pannexin 200:207 arg1 family 209:214 pannexin family 200:214 pannexin family 200:214 Pannexin 1 (PANX1), one of the members of pannexin family, is a channel protein. 15044018 2 0 theme diabetes 468:475 arg1 mellitus 477:484 diabetes mellitus 468:484 diabetes mellitus 468:484 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 1 1 from mice 402:405 arg1 population 340:349 a prevalent population 328:349 a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice 328:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 2 2 theme subcellular 502:512 arg1 localization 514:525 subcellular localization 502:525 subcellular localization 502:525 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 1 3 theme pathogenic 354:363 arg1 CD8 365:367 pathogenic CD8 354:367 pathogenic CD8 354:367 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 0 4 theme embedded 100:107 arg1 glycoprotein 87:98 a glycoprotein 85:98 a glycoprotein embedded in the endoplasmic reticulum membrane 85:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 0 4 theme embedded 100:107 arg1 protein 49:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 1 5 theme CD8 365:367 arg1 cells 374:378 pathogenic CD8(+) T cells 354:378 pathogenic CD8(+) T cells in non-obese diabetic mice 354:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 0 6 theme endoplasmic 116:126 arg1 reticulum 128:136 the endoplasmic reticulum 112:136 the endoplasmic reticulum membrane 112:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 2 7 from roles 443:447 arg1 mellitus 477:484 diabetes mellitus 468:484 diabetes mellitus 468:484 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 2 8 theme membrane 531:538 arg1 topography 540:549 membrane topography 531:549 membrane topography 531:549 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 1 9 from cells 374:378 arg1 mice 402:405 non-obese diabetic mice 383:405 non-obese diabetic mice 383:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 0 10 gly glycoprotein 87:98 arg1 glycoprotein 87:98 a glycoprotein 85:98 a glycoprotein embedded in the endoplasmic reticulum membrane 85:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 0 10 gly glycoprotein 87:98 arg1 protein 49:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 3 11 gly glycoprotein 589:600 arg1 IGRP 579:582 IGRP 579:582 IGRP 579:582 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 3 11 gly glycoprotein 589:600 arg1 glycoprotein 589:600 a glycoprotein 587:600 a glycoprotein 587:600 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 0 12 theme islet-specific 4:17 arg1 protein 49:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 0 12 theme islet-specific 4:17 arg1 glycoprotein 87:98 a glycoprotein 85:98 a glycoprotein embedded in the endoplasmic reticulum membrane 85:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 3 13 theme proteasome 721:730 arg1 pathway 732:738 the proteasome pathway 717:738 the proteasome pathway that generates the major histocompatibility complex class I-presented peptides 717:817 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 1 14 theme cells 374:378 arg1 population 340:349 a prevalent population 328:349 a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice 328:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 3 15 theme endoplasmic 615:625 arg1 reticulum 627:635 the endoplasmic reticulum 611:635 the endoplasmic reticulum 611:635 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 1 16 theme known 219:223 arg1 activity 235:242 no known catalytic activity 216:242 no known catalytic activity 216:242 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 0 17 theme glucose-6-phosphatase-related 19:47 arg1 protein 49:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein 0:55 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 0 17 theme glucose-6-phosphatase-related 19:47 arg1 glycoprotein 87:98 a glycoprotein 85:98 a glycoprotein embedded in the endoplasmic reticulum membrane 85:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 0 18 theme reticulum 128:136 arg1 membrane 138:145 the endoplasmic reticulum membrane 112:145 the endoplasmic reticulum membrane 112:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 1 19 theme T 372:372 arg1 cells 374:378 pathogenic CD8(+) T cells 354:378 pathogenic CD8(+) T cells in non-obese diabetic mice 354:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 20 theme catalytic 225:233 arg1 activity 235:242 no known catalytic activity 216:242 no known catalytic activity 216:242 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 2 21 theme protein 457:463 arg1 roles 443:447 the potential roles 429:447 the potential roles of this protein in diabetes mellitus 429:484 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 1 22 theme peptide 296:302 arg1 autoantigen 304:314 the peptide autoantigen 292:314 the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice 292:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 23 contain has 212:214 arg1 IGRP 206:209 IGRP 206:209 IGRP 206:209 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 23 contain has 212:214 arg2 activity 235:242 no known catalytic activity 216:242 no known catalytic activity 216:242 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 23 contain has 212:214 arg1 protein 197:203 The islet-specific glucose-6-phosphatase-related protein 148:203 The islet-specific glucose-6-phosphatase-related protein (IGRP) 148:210 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 24 theme autoantigen 304:314 arg1 it 272:273 it 272:273 it 272:273 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 24 theme autoantigen 304:314 arg1 source 282:287 the source 278:287 the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice 278:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 3 25 theme histocompatibility 765:782 arg1 class 792:796 major histocompatibility complex class 759:796 the major histocompatibility complex class I-presented peptides 755:817 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 3 26 theme major 759:763 arg1 class 792:796 major histocompatibility complex class 759:796 the major histocompatibility complex class I-presented peptides 755:817 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 1 27 theme non-obese 383:391 arg1 mice 402:405 non-obese diabetic mice 383:405 non-obese diabetic mice 383:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 28 from population 340:349 arg1 mice 402:405 non-obese diabetic mice 383:405 non-obese diabetic mice 383:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 2 29 theme IGRP 560:563 arg1 localization 514:525 subcellular localization 502:525 subcellular localization 502:525 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 2 29 theme IGRP 560:563 arg1 topography 540:549 membrane topography 531:549 membrane topography 531:549 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 1 30 theme diabetic 393:400 arg1 mice 402:405 non-obese diabetic mice 383:405 non-obese diabetic mice 383:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 2 31 theme human 554:558 arg1 IGRP 560:563 human IGRP 554:563 human IGRP 554:563 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 3 32 theme transmembrane 645:657 arg1 domains 659:665 nine transmembrane domains 640:665 nine transmembrane domains 640:665 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 3 33 theme class 792:796 arg1 peptides 810:817 the major histocompatibility complex class I-presented peptides 755:817 the major histocompatibility complex class I-presented peptides 755:817 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 3 34 theme I-presented 798:808 arg1 peptides 810:817 the major histocompatibility complex class I-presented peptides 755:817 the major histocompatibility complex class I-presented peptides 755:817 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 0 35 from embedded 100:107 arg1 membrane 138:145 the endoplasmic reticulum membrane 112:145 the endoplasmic reticulum membrane 112:145 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 1 36 theme islet-specific 152:165 arg1 IGRP 206:209 IGRP 206:209 IGRP 206:209 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 36 theme islet-specific 152:165 arg1 protein 197:203 The islet-specific glucose-6-phosphatase-related protein 148:203 The islet-specific glucose-6-phosphatase-related protein (IGRP) 148:210 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 0 37 from membrane 138:145 arg1 embedded 100:107 embedded 100:107 embedded 100:107 The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. 15044018 2 38 theme potential 433:441 arg1 roles 443:447 the potential roles 429:447 the potential roles of this protein in diabetes mellitus 429:484 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15044018 1 39 theme prevalent 330:338 arg1 population 340:349 a prevalent population 328:349 a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice 328:405 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 3 40 theme complex 784:790 arg1 class 792:796 major histocompatibility complex class 759:796 the major histocompatibility complex class I-presented peptides 755:817 We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. 15044018 1 41 theme glucose-6-phosphatase-related 167:195 arg1 IGRP 206:209 IGRP 206:209 IGRP 206:209 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 1 41 theme glucose-6-phosphatase-related 167:195 arg1 protein 197:203 The islet-specific glucose-6-phosphatase-related protein 148:203 The islet-specific glucose-6-phosphatase-related protein (IGRP) 148:210 The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. 15044018 2 42 dep localization 514:525 arg1 the 498:500 the 498:500 the 498:500 To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. 15728350 1 0 theme Ig 134:135 arg1 IgSF 150:153 IgSF 150:153 IgSF 150:153 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 1 0 theme Ig 134:135 arg1 superfamily 137:147 the Ig superfamily 130:147 the Ig superfamily (IgSF) 130:154 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 2 1 with complex 350:356 arg1 alphaLbeta2 413:423 alphaLbeta2 413:423 alphaLbeta2 413:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 1 with complex 350:356 arg1 domain 403:408 the ligand-binding domain 384:408 the ligand-binding domain of alphaLbeta2 384:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 1 with complex 350:356 arg1 domain 376:381 the inserted domain 363:381 the inserted domain 363:381 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 2 from structure 300:308 arg1 complex 350:356 complex 350:356 complex with the inserted domain, the ligand-binding domain of alphaLbeta2 350:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 3 theme ICAM-3 317:322 arg1 D1 343:344 D1 343:344 D1 343:344 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 3 theme ICAM-3 317:322 arg1 domain 335:340 the ICAM-3 N-terminal domain 313:340 the ICAM-3 N-terminal domain (D1) 313:345 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 0 4 theme binding 71:77 arg1 alphaLbeta2 110:120 integrin alphaLbeta2 101:120 integrin alphaLbeta2 101:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 0 4 theme binding 71:77 arg1 domains 79:85 the binding domains 67:85 the binding domains of ICAM-3 and integrin alphaLbeta2 67:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 0 4 theme binding 71:77 arg1 ICAM-3 90:95 ICAM-3 90:95 ICAM-3 90:95 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 2 5 theme N-terminal 324:333 arg1 D1 343:344 D1 343:344 D1 343:344 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 5 theme N-terminal 324:333 arg1 domain 335:340 the ICAM-3 N-terminal domain 313:340 the ICAM-3 N-terminal domain (D1) 313:345 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 0 6 theme ICAM-3 90:95 arg1 alphaLbeta2 110:120 integrin alphaLbeta2 101:120 integrin alphaLbeta2 101:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 0 6 theme ICAM-3 90:95 arg1 domains 79:85 the binding domains 67:85 the binding domains of ICAM-3 and integrin alphaLbeta2 67:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 0 6 theme ICAM-3 90:95 arg1 ICAM-3 90:95 ICAM-3 90:95 ICAM-3 90:95 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 4 7 theme alphaLbeta2-binding 788:806 arg1 interfaces 808:817 the alphaLbeta2-binding interfaces 784:817 the alphaLbeta2-binding interfaces 784:817 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 2 8 theme inserted 367:374 arg1 alphaLbeta2 413:423 alphaLbeta2 413:423 alphaLbeta2 413:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 8 theme inserted 367:374 arg1 domain 403:408 the ligand-binding domain 384:408 the ligand-binding domain of alphaLbeta2 384:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 8 theme inserted 367:374 arg1 domain 376:381 the inserted domain 363:381 the inserted domain 363:381 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 5 9 theme interfaces 880:889 arg1 periphery 863:871 the periphery 859:871 the periphery of the interfaces 859:889 Variation in composition of glycans on the periphery of the interfaces influences on-rate. 15728350 0 10 theme atomic 3:8 arg1 view 21:24 An atomic resolution view 0:24 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 0:121 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 2 11 theme 1.65-A-resolution 274:290 arg1 structure 300:308 a 1.65-A-resolution crystal structure 272:308 a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2 272:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 4 12 theme different 641:649 arg1 off-rates 651:659 The markedly different off-rates 628:659 The markedly different off-rates of ICAM-1, -2, and -3 628:681 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 2 13 theme ligand-binding 388:401 arg1 alphaLbeta2 413:423 alphaLbeta2 413:423 alphaLbeta2 413:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 13 theme ligand-binding 388:401 arg1 domain 403:408 the ligand-binding domain 384:408 the ligand-binding domain of alphaLbeta2 384:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 13 theme ligand-binding 388:401 arg1 domain 376:381 the inserted domain 363:381 the inserted domain 363:381 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 1 14 theme leukocyte 230:238 arg1 alphaLbeta2 249:259 the leukocyte integrin alphaLbeta2 226:259 the leukocyte integrin alphaLbeta2 226:259 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 4 15 theme ICAM-1 664:669 arg1 off-rates 651:659 The markedly different off-rates 628:659 The markedly different off-rates of ICAM-1, -2, and -3 628:681 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 1 16 theme integrin 240:247 arg1 alphaLbeta2 249:259 the leukocyte integrin alphaLbeta2 226:259 the leukocyte integrin alphaLbeta2 226:259 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 0 17 theme integrin 101:108 arg1 alphaLbeta2 110:120 integrin alphaLbeta2 101:120 integrin alphaLbeta2 101:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 1 18 theme intercellular 157:169 arg1 ICAMs 191:195 ICAMs 191:195 ICAMs 191:195 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 1 18 theme intercellular 157:169 arg1 molecules 180:188 intercellular adhesion molecules 157:188 intercellular adhesion molecules (ICAMs) 157:196 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 0 19 theme resolution 10:19 arg1 view 21:24 An atomic resolution view 0:24 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 0:121 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 3 20 theme subfamily 609:617 arg1 members 619:625 ICAM subfamily members 604:625 ICAM subfamily members 604:625 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 3 21 theme ICAM-3 532:537 arg1 D1 539:540 ICAM-3 D1 532:540 ICAM-3 D1 532:540 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 1 22 theme adhesion 171:178 arg1 ICAMs 191:195 ICAMs 191:195 ICAMs 191:195 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 1 22 theme adhesion 171:178 arg1 molecules 180:188 intercellular adhesion molecules 157:188 intercellular adhesion molecules (ICAMs) 157:196 Within the Ig superfamily (IgSF), intercellular adhesion molecules (ICAMs) form a subfamily that binds the leukocyte integrin alphaLbeta2. 15728350 0 23 theme ICAM 29:32 arg1 recognition 34:44 ICAM recognition 29:44 ICAM recognition 29:44 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 4 24 theme coordination 763:774 arg1 bond 776:779 a metal coordination bond 755:779 a metal coordination bond in the alphaLbeta2-binding interfaces 755:817 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 0 25 theme alphaLbeta2 110:120 arg1 alphaLbeta2 110:120 integrin alphaLbeta2 101:120 integrin alphaLbeta2 101:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 0 25 theme alphaLbeta2 110:120 arg1 domains 79:85 the binding domains 67:85 the binding domains of ICAM-3 and integrin alphaLbeta2 67:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 0 25 theme alphaLbeta2 110:120 arg1 ICAM-3 90:95 ICAM-3 90:95 ICAM-3 90:95 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 4 26 theme metal 757:761 arg1 bond 776:779 a metal coordination bond 755:779 a metal coordination bond in the alphaLbeta2-binding interfaces 755:817 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 2 27 theme alphaLbeta2 413:423 arg1 alphaLbeta2 413:423 alphaLbeta2 413:423 alphaLbeta2 413:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 27 theme alphaLbeta2 413:423 arg1 domain 403:408 the ligand-binding domain 384:408 the ligand-binding domain of alphaLbeta2 384:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 2 27 theme alphaLbeta2 413:423 arg1 domain 376:381 the inserted domain 363:381 the inserted domain 363:381 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 5 28 theme glycans 848:854 arg1 composition 833:843 composition 833:843 composition of glycans 833:854 Variation in composition of glycans on the periphery of the interfaces influences on-rate. 15728350 3 29 theme ICAM 604:607 arg1 members 619:625 ICAM subfamily members 604:625 ICAM subfamily members 604:625 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 4 30 from bond 776:779 arg1 interfaces 808:817 the alphaLbeta2-binding interfaces 784:817 the alphaLbeta2-binding interfaces 784:817 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 0 31 theme recognition 34:44 arg1 view 21:24 An atomic resolution view 0:24 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 0:121 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 3 32 theme inserted 551:558 arg1 domain 560:565 the inserted domain 547:565 the inserted domain 547:565 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 3 33 theme ICAM 479:482 arg1 members 494:500 ICAM subfamily members 479:500 ICAM subfamily members 479:500 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 4 34 theme -2 672:673 arg1 off-rates 651:659 The markedly different off-rates 628:659 The markedly different off-rates of ICAM-1, -2, and -3 628:681 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 3 35 theme D1 539:540 arg1 binding 521:527 the binding 517:527 the binding of ICAM-3 D1 onto the inserted domain 517:565 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 3 36 theme docking 587:593 arg1 mode 595:598 a common docking mode 578:598 a common docking mode for ICAM subfamily members 578:625 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 3 37 theme high-resolution 431:445 arg1 structure 447:455 This high-resolution structure 426:455 This high-resolution structure 426:455 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 4 38 theme residues 732:739 arg1 hydrophobicity 714:727 the hydrophobicity 710:727 the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces 710:817 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 2 39 theme crystal 292:298 arg1 structure 300:308 a 1.65-A-resolution crystal structure 272:308 a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2 272:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 15728350 4 40 theme -3 680:681 arg1 off-rates 651:659 The markedly different off-rates 628:659 The markedly different off-rates of ICAM-1, -2, and -3 628:681 The markedly different off-rates of ICAM-1, -2, and -3 appear to be determined by the hydrophobicity of residues that surround a metal coordination bond in the alphaLbeta2-binding interfaces. 15728350 3 41 theme subfamily 484:492 arg1 members 494:500 ICAM subfamily members 479:500 ICAM subfamily members 479:500 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 0 42 from view 21:24 arg1 complex 51:57 a complex 49:57 a complex between the binding domains of ICAM-3 and integrin alphaLbeta2 49:120 An atomic resolution view of ICAM recognition in a complex between the binding domains of ICAM-3 and integrin alphaLbeta2. 15728350 3 43 theme common 580:585 arg1 mode 595:598 a common docking mode 578:598 a common docking mode for ICAM subfamily members 578:625 This high-resolution structure and comparisons among ICAM subfamily members establish that the binding of ICAM-3 D1 onto the inserted domain represents a common docking mode for ICAM subfamily members. 15728350 5 44 from Variation 820:828 arg1 periphery 863:871 the periphery 859:871 the periphery of the interfaces 859:889 Variation in composition of glycans on the periphery of the interfaces influences on-rate. 15728350 5 44 from Variation 820:828 arg1 composition 833:843 composition 833:843 composition of glycans 833:854 Variation in composition of glycans on the periphery of the interfaces influences on-rate. 15728350 2 45 theme domain 335:340 arg1 structure 300:308 a 1.65-A-resolution crystal structure 272:308 a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2 272:423 We report a 1.65-A-resolution crystal structure of the ICAM-3 N-terminal domain (D1) in complex with the inserted domain, the ligand-binding domain of alphaLbeta2. 2277032 4 0 dep K. 849:850 arg1 al 855:856 et al 852:856 several volunteers [Ito, K. et al. (1984) J. Biochem 824:875 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 4 0 dep K. 849:850 arg1 Biochem 869:875 Biochem 869:875 Biochem 869:875 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 1 1 theme sulfate-polyacrylamide 318:339 arg1 electrophoresis 345:359 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 303:359 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 303:359 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 4 2 theme several 824:830 arg1 volunteers 832:841 several volunteers 824:841 several volunteers [Ito, K. et al. (1984) J. Biochem 824:875 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 10 3 from variations 1215:1224 arg1 differences 1258:1268 differences 1258:1268 differences in the content of sialic acid 1258:1298 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 10 3 from variations 1215:1224 arg1 structure 1241:1249 the primary structure 1229:1249 the primary structure 1229:1249 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 1 4 dep state 284:288 arg1 judged 293:298 judged 293:298 judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis 293:359 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 1 5 theme gel 341:343 arg1 electrophoresis 345:359 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 303:359 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 303:359 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 7 6 theme genetic 1031:1037 arg1 polymorphism 1039:1050 genetic polymorphism 1031:1050 genetic polymorphism of deoxyribonuclease I in human urine previously reported 1031:1108 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 0 7 theme deoxyribonuclease 107:123 arg1 multiplicity 85:96 multiplicity 85:96 multiplicity 85:96 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 2277032 0 7 theme deoxyribonuclease 107:123 arg1 purification 49:60 purification 49:60 purification 49:60 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 2277032 0 7 theme deoxyribonuclease 107:123 arg1 characterization 63:78 characterization 63:78 characterization 63:78 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 2277032 3 8 theme bovine 638:643 arg1 species 655:661 bovine and other species 638:661 bovine and other species 638:661 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 2 9 gly glycoprotein 391:402 arg1 enzyme 366:371 The enzyme 362:371 The enzyme 362:371 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 9 gly glycoprotein 391:402 arg1 glycoprotein 391:402 a glycoprotein 389:402 a glycoprotein 389:402 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 1 10 theme column 243:248 arg1 chromatographies 250:265 column chromatographies 243:265 column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis 243:359 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 2 11 theme acid 479:482 arg1 residues 484:491 2 sialic acid residues 470:491 2 sialic acid residues per molecule 470:504 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 10 12 theme sialic 1288:1293 arg1 acid 1295:1298 sialic acid 1288:1298 sialic acid 1288:1298 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 1 13 theme chromatographies 250:265 arg1 series 233:238 a series 231:238 a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis 231:359 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 2 14 theme sialic 472:477 arg1 residues 484:491 2 sialic acid residues 470:491 2 sialic acid residues per molecule 470:504 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 4 15 theme human 797:801 arg1 urine 803:807 human urine 797:807 human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem 797:875 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 10 16 theme primary 1233:1239 arg1 structure 1241:1249 the primary structure 1229:1249 the primary structure 1229:1249 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 4 17 theme single 718:723 arg1 individual 725:734 a single individual 716:734 a single individual 716:734 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 7 18 from urine 1084:1088 arg1 polymorphism 1039:1050 genetic polymorphism 1031:1050 genetic polymorphism of deoxyribonuclease I in human urine previously reported 1031:1108 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 7 19 dep existence 1018:1026 arg1 Hum 1135:1137 Hum 1135:1137 Hum 1135:1137 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 7 19 dep existence 1018:1026 arg1 1989 1129:1132 1989 1129:1132 1989 1129:1132 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 7 19 dep existence 1018:1026 arg1 K. 1118:1119 K. 1118:1119 K. 1118:1119 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 7 20 theme reported 1101:1108 arg1 polymorphism 1039:1050 genetic polymorphism 1031:1050 genetic polymorphism of deoxyribonuclease I in human urine previously reported 1031:1108 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 3 21 from species 655:661 arg1 I 631:631 pancreatic deoxyribonuclease I 602:631 pancreatic deoxyribonuclease I from bovine and other species 602:661 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 22 theme acid 528:531 arg1 similar 583:589 similar 583:589 similar 583:589 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 22 theme acid 528:531 arg1 sequence 533:540 The N-terminal amino acid sequence 507:540 The N-terminal amino acid sequence up to the 27th residue of the enzyme 507:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 0 23 theme Human 0:4 arg1 deoxyribonuclease 30:46 Human genetically polymorphic deoxyribonuclease 0:46 Human genetically polymorphic deoxyribonuclease 0:46 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 2277032 4 24 attach derived 703:709 arg1 individual 725:734 a single individual 716:734 a single individual 716:734 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 4 24 attach derived 703:709 arg2 enzyme 696:701 the enzyme 692:701 the enzyme derived from a single individual 692:734 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 10 25 from differences 1258:1268 arg1 content 1277:1283 the content 1273:1283 the content of sialic acid 1273:1298 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 10 26 theme urine 1186:1190 arg1 enzyme 1192:1197 the urine enzyme 1182:1197 the urine enzyme 1182:1197 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 1 27 theme male 195:198 arg1 urine 172:176 the urine 168:176 the urine of a 46-year-old male (a single individual) 168:220 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 10 28 theme acid 1295:1298 arg1 content 1277:1283 the content 1273:1283 the content of sialic acid 1273:1298 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 3 29 theme N-terminal 511:520 arg1 similar 583:589 similar 583:589 similar 583:589 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 29 theme N-terminal 511:520 arg1 sequence 533:540 The N-terminal amino acid sequence 507:540 The N-terminal amino acid sequence up to the 27th residue of the enzyme 507:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 30 dep residue 557:563 arg1 up 542:543 up 542:543 up 542:543 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 0 31 theme polymorphic 18:28 arg1 deoxyribonuclease 30:46 Human genetically polymorphic deoxyribonuclease 0:46 Human genetically polymorphic deoxyribonuclease 0:46 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 2277032 1 32 theme homogeneous 272:282 arg1 state 284:288 a homogeneous state 270:288 a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis 270:359 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 3 33 theme amino 522:526 arg1 similar 583:589 similar 583:589 similar 583:589 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 33 theme amino 522:526 arg1 sequence 533:540 The N-terminal amino acid sequence 507:540 The N-terminal amino acid sequence up to the 27th residue of the enzyme 507:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 7 34 theme deoxyribonuclease 1055:1071 arg1 I 1073:1073 deoxyribonuclease I 1055:1073 deoxyribonuclease I in human urine 1055:1088 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 4 35 theme deoxyribonuclease 763:779 arg1 I 781:781 deoxyribonuclease I 763:781 deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem 763:875 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 6 36 theme pI 968:969 arg1 values 971:976 different pI values 958:976 different pI values 958:976 The purified enzyme was found to consist of multiple forms with different pI values. 2277032 1 37 theme single 203:208 arg1 male 195:198 male 195:198 male 195:198 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 1 37 theme single 203:208 arg1 individual 210:219 a single individual 201:219 a single individual 201:219 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 4 38 dep volunteers 832:841 arg1 1984 860:863 1984 860:863 1984 860:863 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 4 38 dep volunteers 832:841 arg1 K. 849:850 K. 849:850 K. 849:850 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 7 39 theme I 1073:1073 arg1 polymorphism 1039:1050 genetic polymorphism 1031:1050 genetic polymorphism of deoxyribonuclease I in human urine previously reported 1031:1108 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 3 40 theme 27th 552:555 arg1 residue 557:563 the 27th residue 548:563 the 27th residue of the enzyme 548:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 40 theme 27th 552:555 arg1 enzyme 572:577 the enzyme 568:577 the enzyme 568:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 6 41 theme multiple 938:945 arg1 forms 947:951 multiple forms 938:951 multiple forms 938:951 The purified enzyme was found to consist of multiple forms with different pI values. 2277032 7 42 theme polymorphism 1039:1050 arg1 existence 1018:1026 the existence 1014:1026 the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum 1014:1137 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 0 43 dep purification 49:60 arg1 deoxyribonuclease 30:46 Human genetically polymorphic deoxyribonuclease 0:46 Human genetically polymorphic deoxyribonuclease 0:46 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 2277032 7 44 from I 1073:1073 arg1 urine 1084:1088 human urine 1078:1088 human urine 1078:1088 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 3 45 theme deoxyribonuclease 613:629 arg1 I 631:631 pancreatic deoxyribonuclease I 602:631 pancreatic deoxyribonuclease I from bovine and other species 602:661 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 1 46 theme deoxyribonuclease 130:146 arg1 I 148:148 A deoxyribonuclease I 128:148 A deoxyribonuclease I 128:148 A deoxyribonuclease I was purified from the urine of a 46-year-old male (a single individual) by using a series of column chromatographies to a homogeneous state as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2277032 10 47 theme enzyme 1192:1197 arg1 multiplicity 1166:1177 This multiplicity 1161:1177 This multiplicity of the urine enzyme 1161:1197 This multiplicity of the urine enzyme might be due to variations in the primary structure and/or differences in the content of sialic acid. 2277032 4 48 theme catalytic 668:676 arg1 properties 678:687 The catalytic properties 664:687 The catalytic properties of the enzyme derived from a single individual 664:734 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 4 49 theme enzyme 696:701 arg1 properties 678:687 The catalytic properties 664:687 The catalytic properties of the enzyme derived from a single individual 664:734 The catalytic properties of the enzyme derived from a single individual closely resembled those of deoxyribonuclease I purified from human urine collected from several volunteers [Ito, K. et al. (1984) J. Biochem. 2277032 7 50 dep K. 1118:1119 arg1 al 1124:1125 et al 1121:1125 the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum 1014:1137 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 2 51 contain containing 405:414 arg2 mannose 442:448 10 mannose 439:448 10 mannose 439:448 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 51 contain containing 405:414 arg1 enzyme 366:371 The enzyme 362:371 The enzyme 362:371 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 51 contain containing 405:414 arg1 glycoprotein 391:402 a glycoprotein 389:402 a glycoprotein 389:402 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 51 contain containing 405:414 arg2 residues 484:491 2 sialic acid residues 470:491 2 sialic acid residues per molecule 470:504 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 51 contain containing 405:414 arg2 fucose 418:423 1 fucose 416:423 1 fucose 416:423 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 51 contain containing 405:414 arg2 galactose 428:436 7 galactose 426:436 7 galactose 426:436 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 2 51 contain containing 405:414 arg2 glucosamine 453:463 6 glucosamine 451:463 6 glucosamine 451:463 The enzyme was found to be a glycoprotein, containing 1 fucose, 7 galactose, 10 mannose, 6 glucosamine, and 2 sialic acid residues per molecule. 2277032 3 52 theme enzyme 572:577 arg1 residue 557:563 the 27th residue 548:563 the 27th residue of the enzyme 548:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 3 52 theme enzyme 572:577 arg1 enzyme 572:577 the enzyme 568:577 the enzyme 568:577 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 7 53 with compatible 998:1007 arg1 existence 1018:1026 the existence 1014:1026 the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum 1014:1137 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 3 54 theme pancreatic 602:611 arg1 I 631:631 pancreatic deoxyribonuclease I 602:631 pancreatic deoxyribonuclease I from bovine and other species 602:661 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 6 55 theme different 958:966 arg1 values 971:976 different pI values 958:976 different pI values 958:976 The purified enzyme was found to consist of multiple forms with different pI values. 2277032 6 56 theme purified 898:905 arg1 enzyme 907:912 The purified enzyme 894:912 The purified enzyme 894:912 The purified enzyme was found to consist of multiple forms with different pI values. 2277032 7 57 from polymorphism 1039:1050 arg1 urine 1084:1088 human urine 1078:1088 human urine 1078:1088 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 7 58 theme human 1078:1082 arg1 urine 1084:1088 human urine 1078:1088 human urine 1078:1088 These findings are compatible with the existence of genetic polymorphism of deoxyribonuclease I in human urine previously reported [Kishi, K. et al. (1989) Hum. 2277032 3 59 theme other 649:653 arg1 species 655:661 bovine and other species 638:661 bovine and other species 638:661 The N-terminal amino acid sequence up to the 27th residue of the enzyme was similar to that of pancreatic deoxyribonuclease I from bovine and other species. 2277032 0 60 theme urine 101:105 arg1 deoxyribonuclease 107:123 urine deoxyribonuclease I 101:125 urine deoxyribonuclease I 101:125 Human genetically polymorphic deoxyribonuclease: purification, characterization, and multiplicity of urine deoxyribonuclease I. 14718370 2 0 theme histidine 311:319 arg1 residues 321:328 six carboxyl terminal histidine residues 289:328 six carboxyl terminal histidine residues 289:328 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 9 1 theme fluorescent 1378:1388 arg1 protein 1390:1396 green fluorescent protein 1372:1396 green fluorescent protein fused to the Cys subdomains 1372:1424 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 3 2 theme secreted 475:482 arg1 subdomains 488:497 secreted Cys subdomains 475:497 secreted Cys subdomains 475:497 Under nonreducing conditions, secreted Cys subdomains were monomers, indicating the absence of interchain disulfide bonds. 14718370 1 3 theme domain 160:165 arg1 role 122:125 the role 118:125 the role of this highly conserved protein domain in mucin biosynthesis 118:187 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 6 4 theme Lectin 817:822 arg1 binding 824:830 Lectin binding 817:830 Lectin binding 817:830 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 5 5 theme mannose-specific 791:806 arg1 lectins 808:814 mannose-specific lectins 791:814 mannose-specific lectins 791:814 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 1 6 theme mucin 170:174 arg1 biosynthesis 176:187 mucin biosynthesis 170:187 mucin biosynthesis 170:187 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 6 7 theme C-mannosylation 906:920 arg1 WXXW 937:940 the putative C-mannosylation acceptor motif WXXW 893:940 the putative C-mannosylation acceptor motif WXXW 893:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 9 8 from presence 1453:1460 arg1 ER 1487:1488 the ER 1483:1488 the ER of CHO-Lec35.1 cells 1483:1509 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 8 9 theme unmutated 1206:1214 arg1 line 1278:1281 a C-mannosylation-defective cell line 1245:1281 a C-mannosylation-defective cell line 1245:1281 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 9 theme unmutated 1206:1214 arg1 domain 1216:1221 the unmutated domain 1202:1221 the unmutated domain in CHO-Lec35.1 cells 1202:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 7 10 theme pulse-chase 1023:1033 arg1 experiments 1035:1045 pulse-chase experiments 1023:1045 pulse-chase experiments 1023:1045 As judged by pulse-chase experiments, C-mannosylation occurred very early during the domain biosynthesis, likely in the endoplasmic reticulum (ER). 14718370 11 11 theme export 1816:1821 arg1 aspect 1803:1808 some aspect 1798:1808 some aspect of ER export during mucin biosynthesis 1798:1847 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 11 12 theme subdomains 1776:1785 arg1 folding 1757:1763 proper folding 1750:1763 proper folding of the Cys subdomains 1750:1785 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 3 13 theme interchain 540:549 arg1 bonds 561:565 interchain disulfide bonds 540:565 interchain disulfide bonds 540:565 Under nonreducing conditions, secreted Cys subdomains were monomers, indicating the absence of interchain disulfide bonds. 14718370 8 14 theme WXXW 1174:1177 arg1 motif 1179:1183 the WXXW motif 1170:1183 the WXXW motif 1170:1183 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 6 15 theme residue 882:888 arg1 mutation 849:856 mutation 849:856 mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW 849:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 6 16 theme first 865:869 arg1 residue 882:888 the first tryptophan residue 861:888 the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW 861:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 2 17 theme Cys3 228:231 arg1 subdomains 233:242 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains 190:242 subdomains 233:242 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 9 18 theme Cys 1465:1467 arg1 subdomains 1469:1478 Cys subdomains 1465:1478 Cys subdomains 1465:1478 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 8 19 theme cell 1273:1276 arg1 domain 1216:1221 the unmutated domain 1202:1221 the unmutated domain in CHO-Lec35.1 cells 1202:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 19 theme cell 1273:1276 arg1 line 1278:1281 a C-mannosylation-defective cell line 1245:1281 a C-mannosylation-defective cell line 1245:1281 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 3 20 theme bonds 561:565 arg1 absence 529:535 the absence 525:535 the absence of interchain disulfide bonds 525:565 Under nonreducing conditions, secreted Cys subdomains were monomers, indicating the absence of interchain disulfide bonds. 14718370 4 21 theme noncovalent 651:661 arg1 interactions 663:674 very weak noncovalent interactions 641:674 very weak noncovalent interactions 641:674 Cross-linking studies suggested the domains are able to interact through very weak noncovalent interactions. 14718370 5 22 theme consistent 714:723 arg1 r 711:711 r 711:711 r 711:711 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 5 22 theme consistent 714:723 arg1 M 709:709 apparent M 700:709 apparent M(r) consistent with the absence of N- and O-glycans 700:760 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 1 23 theme conserved 142:150 arg1 domain 160:165 this highly conserved protein domain 130:165 this highly conserved protein domain 130:165 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 8 24 from cells 1238:1242 arg1 expression 1188:1197 expression 1188:1197 expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, 1188:1282 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 24 from cells 1238:1242 arg1 Mutation 1158:1165 Mutation 1158:1165 Mutation of the WXXW motif 1158:1183 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 9 25 theme CHO-Lec35.1 1493:1503 arg1 cells 1505:1509 CHO-Lec35.1 cells 1493:1509 CHO-Lec35.1 cells 1493:1509 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 6 26 theme motif 931:935 arg1 WXXW 937:940 the putative C-mannosylation acceptor motif WXXW 893:940 the putative C-mannosylation acceptor motif WXXW 893:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 6 27 from WXXW 937:940 arg1 mutation 849:856 mutation 849:856 mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW 849:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 1 28 theme Cys 77:79 arg1 subdomains 81:90 recombinant Cys subdomains 65:90 recombinant Cys subdomains 65:90 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 9 29 attach presence 1453:1460 arg2 subdomains 1469:1478 Cys subdomains 1465:1478 Cys subdomains 1465:1478 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 9 29 attach presence 1453:1460 arg1 ER 1487:1488 the ER 1483:1488 the ER of CHO-Lec35.1 cells 1483:1509 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 8 30 theme Cys 1335:1337 arg1 subdomains 1339:1348 the corresponding Cys subdomains 1317:1348 the corresponding Cys subdomains 1317:1348 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 0 31 theme MUC5AC 19:24 arg1 C-Mannosylation 0:14 C-Mannosylation 0:14 C-Mannosylation of MUC5AC and MUC5B Cys subdomains. 0:50 C-Mannosylation of MUC5AC and MUC5B Cys subdomains. 14718370 9 32 theme same 1532:1535 arg1 domains 1537:1543 the same domains 1528:1543 the same domains expressed in CHO-K1 cells 1528:1569 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 2 33 from subdomains 233:242 arg1 MUC5B 258:262 MUC5B 258:262 MUC5B 258:262 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 33 from subdomains 233:242 arg1 MUC5AC 247:252 MUC5AC 247:252 MUC5AC 247:252 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 9 34 theme protein 1390:1396 arg1 imaging 1361:1367 Live cell imaging 1351:1367 Live cell imaging of green fluorescent protein fused to the Cys subdomains 1351:1424 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 0 35 theme MUC5B 30:34 arg1 C-Mannosylation 0:14 C-Mannosylation 0:14 C-Mannosylation of MUC5AC and MUC5B Cys subdomains. 0:50 C-Mannosylation of MUC5AC and MUC5B Cys subdomains. 14718370 2 36 theme terminal 302:309 arg1 residues 321:328 six carboxyl terminal histidine residues 289:328 six carboxyl terminal histidine residues 289:328 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 9 37 theme CHO-K1 1558:1563 arg1 cells 1565:1569 CHO-K1 cells 1558:1569 CHO-K1 cells 1558:1569 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 10 38 theme MUC5AC 1642:1647 arg1 C-mannosylated 1663:1676 C-mannosylated 1663:1676 C-mannosylated 1663:1676 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 38 theme MUC5AC 1642:1647 arg1 MUC5B 1653:1657 MUC5B 1653:1657 MUC5B 1653:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 38 theme MUC5AC 1642:1647 arg1 MUC5AC 1642:1647 MUC5AC 1642:1647 MUC5AC 1642:1647 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 38 theme MUC5AC 1642:1647 arg1 subdomains 1628:1637 the Cys subdomains 1620:1637 the Cys subdomains of MUC5AC and MUC5B 1620:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 39 theme respective 1687:1696 arg1 motifs 1703:1708 their respective WXXW motifs 1681:1708 their respective WXXW motifs 1681:1708 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 0 40 dep MUC5AC 19:24 arg1 subdomains 40:49 Cys subdomains 36:49 Cys subdomains 36:49 C-Mannosylation of MUC5AC and MUC5B Cys subdomains. 14718370 6 41 from mutation 849:856 arg1 WXXW 937:940 the putative C-mannosylation acceptor motif WXXW 893:940 the putative C-mannosylation acceptor motif WXXW 893:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 3 42 theme nonreducing 451:461 arg1 conditions 463:472 nonreducing conditions 451:472 nonreducing conditions 451:472 Under nonreducing conditions, secreted Cys subdomains were monomers, indicating the absence of interchain disulfide bonds. 14718370 8 43 theme motif 1179:1183 arg1 expression 1188:1197 expression 1188:1197 expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, 1188:1282 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 43 theme motif 1179:1183 arg1 Mutation 1158:1165 Mutation 1158:1165 Mutation of the WXXW motif 1158:1183 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 9 44 theme Live 1351:1354 arg1 imaging 1361:1367 Live cell imaging 1351:1367 Live cell imaging of green fluorescent protein fused to the Cys subdomains 1351:1424 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 8 45 from Mutation 1158:1165 arg1 cells 1238:1242 CHO-Lec35.1 cells 1226:1242 CHO-Lec35.1 cells 1226:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 11 46 theme ER 1813:1814 arg1 export 1816:1821 ER export 1813:1821 ER export 1813:1821 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 3 47 theme Cys 484:486 arg1 subdomains 488:497 secreted Cys subdomains 475:497 secreted Cys subdomains 475:497 Under nonreducing conditions, secreted Cys subdomains were monomers, indicating the absence of interchain disulfide bonds. 14718370 9 48 theme green 1372:1376 arg1 protein 1390:1396 green fluorescent protein 1372:1396 green fluorescent protein fused to the Cys subdomains 1372:1424 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 10 49 theme MUC5B 1653:1657 arg1 C-mannosylated 1663:1676 C-mannosylated 1663:1676 C-mannosylated 1663:1676 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 49 theme MUC5B 1653:1657 arg1 MUC5B 1653:1657 MUC5B 1653:1657 MUC5B 1653:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 49 theme MUC5B 1653:1657 arg1 MUC5AC 1642:1647 MUC5AC 1642:1647 MUC5AC 1642:1647 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 49 theme MUC5B 1653:1657 arg1 subdomains 1628:1637 the Cys subdomains 1620:1637 the Cys subdomains of MUC5AC and MUC5B 1620:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 50 theme WXXW 1698:1701 arg1 motifs 1703:1708 their respective WXXW motifs 1681:1708 their respective WXXW motifs 1681:1708 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 1 51 theme protein 152:158 arg1 domain 160:165 this highly conserved protein domain 130:165 this highly conserved protein domain 130:165 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 10 52 from C-mannosylated 1663:1676 arg1 motifs 1703:1708 their respective WXXW motifs 1681:1708 their respective WXXW motifs 1681:1708 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 7 53 gly C-mannosylation 1048:1062 arg2 reticulum 1142:1150 the endoplasmic reticulum 1126:1150 the endoplasmic reticulum (ER) 1126:1155 As judged by pulse-chase experiments, C-mannosylation occurred very early during the domain biosynthesis, likely in the endoplasmic reticulum (ER). 14718370 7 53 gly C-mannosylation 1048:1062 arg2 ER 1153:1154 ER 1153:1154 ER 1153:1154 As judged by pulse-chase experiments, C-mannosylation occurred very early during the domain biosynthesis, likely in the endoplasmic reticulum (ER). 14718370 6 54 theme acceptor 922:929 arg1 WXXW 937:940 the putative C-mannosylation acceptor motif WXXW 893:940 the putative C-mannosylation acceptor motif WXXW 893:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 9 55 theme Cys 1411:1413 arg1 subdomains 1415:1424 the Cys subdomains 1407:1424 the Cys subdomains 1407:1424 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 6 56 theme putative 897:904 arg1 WXXW 937:940 the putative C-mannosylation acceptor motif WXXW 893:940 the putative C-mannosylation acceptor motif WXXW 893:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 11 57 theme proper 1750:1755 arg1 folding 1757:1763 proper folding 1750:1763 proper folding of the Cys subdomains 1750:1785 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 8 58 theme domain 1216:1221 arg1 expression 1188:1197 expression 1188:1197 expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, 1188:1282 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 58 theme domain 1216:1221 arg1 Mutation 1158:1165 Mutation 1158:1165 Mutation of the WXXW motif 1158:1183 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 59 from expression 1188:1197 arg1 cells 1238:1242 CHO-Lec35.1 cells 1226:1242 CHO-Lec35.1 cells 1226:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 9 60 theme increased 1443:1451 arg1 presence 1453:1460 increased presence 1443:1460 increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells 1443:1509 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 6 61 theme tryptophan 871:880 arg1 residue 882:888 the first tryptophan residue 861:888 the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW 861:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 8 62 theme CHO-Lec35.1 1226:1236 arg1 cells 1238:1242 CHO-Lec35.1 cells 1226:1242 CHO-Lec35.1 cells 1226:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 11 63 theme Cys 1772:1774 arg1 subdomains 1776:1785 the Cys subdomains 1768:1785 the Cys subdomains 1768:1785 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 5 64 with consistent 714:723 arg1 absence 734:740 the absence 730:740 the absence of N- and O-glycans 730:760 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 3 65 theme disulfide 551:559 arg1 bonds 561:565 interchain disulfide bonds 540:565 interchain disulfide bonds 540:565 Under nonreducing conditions, secreted Cys subdomains were monomers, indicating the absence of interchain disulfide bonds. 14718370 8 66 from domain 1216:1221 arg1 cells 1238:1242 CHO-Lec35.1 cells 1226:1242 CHO-Lec35.1 cells 1226:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 9 67 theme subdomains 1469:1478 arg1 presence 1453:1460 increased presence 1443:1460 increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells 1443:1509 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 10 68 from motifs 1703:1708 arg1 C-mannosylated 1663:1676 C-mannosylated 1663:1676 C-mannosylated 1663:1676 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 68 from motifs 1703:1708 arg1 MUC5B 1653:1657 MUC5B 1653:1657 MUC5B 1653:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 68 from motifs 1703:1708 arg1 MUC5AC 1642:1647 MUC5AC 1642:1647 MUC5AC 1642:1647 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 68 from motifs 1703:1708 arg1 subdomains 1628:1637 the Cys subdomains 1620:1637 the Cys subdomains of MUC5AC and MUC5B 1620:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 2 69 theme culture 429:435 arg1 medium 437:442 the culture medium 425:442 the culture medium 425:442 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 8 70 theme C-mannosylation-defective 1247:1271 arg1 domain 1216:1221 the unmutated domain 1202:1221 the unmutated domain in CHO-Lec35.1 cells 1202:1242 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 8 70 theme C-mannosylation-defective 1247:1271 arg1 line 1278:1281 a C-mannosylation-defective cell line 1245:1281 a C-mannosylation-defective cell line 1245:1281 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 4 71 theme weak 646:649 arg1 interactions 663:674 very weak noncovalent interactions 641:674 very weak noncovalent interactions 641:674 Cross-linking studies suggested the domains are able to interact through very weak noncovalent interactions. 14718370 10 72 gly C-mannosylated 1663:1676 arg1 C-mannosylated 1663:1676 C-mannosylated 1663:1676 C-mannosylated 1663:1676 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 72 gly C-mannosylated 1663:1676 arg1 MUC5B 1653:1657 MUC5B 1653:1657 MUC5B 1653:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 72 gly C-mannosylated 1663:1676 arg1 MUC5AC 1642:1647 MUC5AC 1642:1647 MUC5AC 1642:1647 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 72 gly C-mannosylated 1663:1676 arg1 subdomains 1628:1637 the Cys subdomains 1620:1637 the Cys subdomains of MUC5AC and MUC5B 1620:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 5 73 theme apparent 700:707 arg1 r 711:711 r 711:711 r 711:711 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 5 73 theme apparent 700:707 arg1 M 709:709 apparent M 700:709 apparent M(r) consistent with the absence of N- and O-glycans 700:760 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 1 74 theme recombinant 65:75 arg1 subdomains 81:90 recombinant Cys subdomains 65:90 recombinant Cys subdomains 65:90 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 2 75 from Cys1 201:204 arg1 MUC5B 258:262 MUC5B 258:262 MUC5B 258:262 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 75 from Cys1 201:204 arg1 MUC5AC 247:252 MUC5AC 247:252 MUC5AC 247:252 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 5 76 contain had 696:698 arg1 domains 688:694 the domains 684:694 the domains 684:694 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 5 76 contain had 696:698 arg2 M 709:709 apparent M 700:709 apparent M(r) consistent with the absence of N- and O-glycans 700:760 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 5 76 contain had 696:698 arg2 r 711:711 r 711:711 r 711:711 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 8 77 theme reduced 1296:1302 arg1 secretion 1304:1312 reduced secretion 1296:1312 reduced secretion of the corresponding Cys subdomains 1296:1348 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 5 78 theme N- 745:746 arg1 absence 734:740 the absence 730:740 the absence of N- and O-glycans 730:760 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 1 79 from role 122:125 arg1 biosynthesis 176:187 mucin biosynthesis 170:187 mucin biosynthesis 170:187 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 2 80 from Cys5 210:213 arg1 MUC5B 258:262 MUC5B 258:262 MUC5B 258:262 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 80 from Cys5 210:213 arg1 MUC5AC 247:252 MUC5AC 247:252 MUC5AC 247:252 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 11 81 gly C-mannosylation 1711:1725 arg1 export 1816:1821 ER export 1813:1821 ER export 1813:1821 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 11 81 gly C-mannosylation 1711:1725 arg1 subdomains 1776:1785 the Cys subdomains 1768:1785 the Cys subdomains 1768:1785 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 5 82 theme O-glycans 752:760 arg1 absence 734:740 the absence 730:740 the absence of N- and O-glycans 730:760 Though the domains had apparent M(r) consistent with the absence of N- and O-glycans, they could be purified with mannose-specific lectins. 14718370 1 83 theme COS-7 95:99 arg1 cells 101:105 COS-7 cells 95:105 COS-7 cells 95:105 We expressed recombinant Cys subdomains in COS-7 cells to examine the role of this highly conserved protein domain in mucin biosynthesis. 14718370 2 84 from Cys1 219:222 arg1 MUC5B 258:262 MUC5B 258:262 MUC5B 258:262 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 84 from Cys1 219:222 arg1 MUC5AC 247:252 MUC5AC 247:252 MUC5AC 247:252 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 9 85 theme cells 1505:1509 arg1 ER 1487:1488 the ER 1483:1488 the ER of CHO-Lec35.1 cells 1483:1509 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 14718370 8 86 theme corresponding 1321:1333 arg1 subdomains 1339:1348 the corresponding Cys subdomains 1317:1348 the corresponding Cys subdomains 1317:1348 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 10 87 theme Cys 1624:1626 arg1 C-mannosylated 1663:1676 C-mannosylated 1663:1676 C-mannosylated 1663:1676 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 87 theme Cys 1624:1626 arg1 MUC5B 1653:1657 MUC5B 1653:1657 MUC5B 1653:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 87 theme Cys 1624:1626 arg1 MUC5AC 1642:1647 MUC5AC 1642:1647 MUC5AC 1642:1647 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 10 87 theme Cys 1624:1626 arg1 subdomains 1628:1637 the Cys subdomains 1620:1637 the Cys subdomains of MUC5AC and MUC5B 1620:1657 Considered together, these studies suggest that the Cys subdomains of MUC5AC and MUC5B are C-mannosylated in their respective WXXW motifs. 14718370 0 88 theme Cys 36:38 arg1 subdomains 40:49 Cys subdomains 36:49 Cys subdomains 36:49 C-Mannosylation of MUC5AC and MUC5B Cys subdomains. 14718370 6 89 theme lectin 994:999 arg1 binding 1001:1007 lectin binding 994:1007 lectin binding 994:1007 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 8 90 theme subdomains 1339:1348 arg1 secretion 1304:1312 reduced secretion 1296:1312 reduced secretion of the corresponding Cys subdomains 1296:1348 Mutation of the WXXW motif or expression of the unmutated domain in CHO-Lec35.1 cells, a C-mannosylation-defective cell line, resulted in reduced secretion of the corresponding Cys subdomains. 14718370 7 91 theme domain 1095:1100 arg1 biosynthesis 1102:1113 the domain biosynthesis 1091:1113 the domain biosynthesis 1091:1113 As judged by pulse-chase experiments, C-mannosylation occurred very early during the domain biosynthesis, likely in the endoplasmic reticulum (ER). 14718370 2 92 theme labeled 391:397 arg1 proteins 399:406 the labeled proteins 387:406 the labeled proteins 387:406 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 93 theme entire 194:199 arg1 Cys1 201:204 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains 190:242 Cys1 201:204 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 93 theme entire 194:199 arg1 each 279:282 each 279:282 each 279:282 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 2 94 theme carboxyl 293:300 arg1 residues 321:328 six carboxyl terminal histidine residues 289:328 six carboxyl terminal histidine residues 289:328 The entire Cys1 and Cys5 and Cys1 and Cys3 subdomains in MUC5AC and MUC5B, respectively, each with six carboxyl terminal histidine residues, were pulse-labeled with [(35)S]cysteine/methionine, and the labeled proteins were examined in the culture medium. 14718370 6 95 from residue 882:888 arg1 WXXW 937:940 the putative C-mannosylation acceptor motif WXXW 893:940 the putative C-mannosylation acceptor motif WXXW 893:940 Lectin binding was prevented by mutation of the first tryptophan residue in the putative C-mannosylation acceptor motif WXXW, indicating that C-mannosylation is responsible for lectin binding. 14718370 11 96 theme mucin 1830:1834 arg1 biosynthesis 1836:1847 mucin biosynthesis 1830:1847 mucin biosynthesis 1830:1847 C-mannosylation is likely required for proper folding of the Cys subdomains and/or for some aspect of ER export during mucin biosynthesis. 14718370 4 97 theme Cross-linking 568:580 arg1 studies 582:588 Cross-linking studies 568:588 Cross-linking studies 568:588 Cross-linking studies suggested the domains are able to interact through very weak noncovalent interactions. 14718370 7 98 theme endoplasmic 1130:1140 arg1 ER 1153:1154 ER 1153:1154 ER 1153:1154 As judged by pulse-chase experiments, C-mannosylation occurred very early during the domain biosynthesis, likely in the endoplasmic reticulum (ER). 14718370 7 98 theme endoplasmic 1130:1140 arg1 reticulum 1142:1150 the endoplasmic reticulum 1126:1150 the endoplasmic reticulum (ER) 1126:1155 As judged by pulse-chase experiments, C-mannosylation occurred very early during the domain biosynthesis, likely in the endoplasmic reticulum (ER). 14718370 9 99 theme cell 1356:1359 arg1 imaging 1361:1367 Live cell imaging 1351:1367 Live cell imaging of green fluorescent protein fused to the Cys subdomains 1351:1424 Live cell imaging of green fluorescent protein fused to the Cys subdomains clearly revealed increased presence of Cys subdomains in the ER of CHO-Lec35.1 cells when compared to the same domains expressed in CHO-K1 cells. 15809306 0 0 theme activation 81:90 arg1 alpha 100:104 human fibroblast activation protein alpha 64:104 human fibroblast activation protein alpha 64:104 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 6 1 theme Glu 927:929 arg1 motif 931:935 the Glu motif 923:935 the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme 923:1034 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 6 2 theme active 1010:1015 arg1 site 1017:1020 the active site 1006:1020 the active site of the enzyme 1006:1034 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 6 2 theme active 1010:1015 arg1 enzyme 1029:1034 the enzyme 1025:1034 the enzyme 1025:1034 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 10 3 dep specificity 1686:1696 arg1 the 1672:1674 the 1672:1674 the 1672:1674 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 0 4 theme fibroblast 70:79 arg1 alpha 100:104 human fibroblast activation protein alpha 64:104 human fibroblast activation protein alpha 64:104 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 8 5 theme catalytic 1354:1362 arg1 efficiency 1364:1373 the catalytic efficiency 1350:1373 the catalytic efficiency 1350:1373 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 10 6 theme clear 1606:1610 arg1 understanding 1612:1624 a clear understanding 1604:1624 a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha 1604:1735 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 6 7 theme crystal 833:839 arg1 structures 841:850 the crystal structures 829:850 the crystal structures of FAPalpha and DPPIV 829:872 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 3 8 theme alpha/beta-hydrolase 486:505 arg1 domain 507:512 an alpha/beta-hydrolase domain 483:512 an alpha/beta-hydrolase domain 483:512 FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. 15809306 1 9 theme tumor 256:260 arg1 growth 262:267 tumor growth 256:267 tumor growth 256:267 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 0 10 theme alpha 100:104 arg1 specificity 49:59 the substrate specificity 35:59 the substrate specificity of human fibroblast activation protein alpha 35:104 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 8 11 theme k 1391:1391 arg1 /K 1397:1398 k(cat)/K 1391:1398 k(cat)/K(m) 1391:1401 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 8 11 theme k 1391:1391 arg1 m 1400:1400 m 1400:1400 m 1400:1400 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 3 12 theme beta-propeller 534:547 arg1 domain 549:554 an eight-bladed beta-propeller domain 518:554 an eight-bladed beta-propeller domain 518:554 FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. 15809306 8 13 theme mutant 1267:1272 arg1 FAPalpha/A657D 1274:1287 the mutant FAPalpha/A657D 1263:1287 the mutant FAPalpha/A657D 1263:1287 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 6 14 dep motif 931:935 arg1 -Glu 946:949 Glu(203)-Glu(204) 938:954 Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV 938:996 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 0 15 theme protein 92:98 arg1 alpha 100:104 human fibroblast activation protein alpha 64:104 human fibroblast activation protein alpha 64:104 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 7 16 from acidity 1105:1111 arg1 pocket 1121:1126 this pocket 1116:1126 this pocket 1116:1126 Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. 15809306 10 17 theme responsible 1656:1666 arg1 determinants 1643:1654 the molecular determinants 1629:1654 the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha 1629:1735 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 8 18 theme dipeptide 1424:1432 arg1 substrates 1434:1443 dipeptide substrates 1424:1443 dipeptide substrates 1424:1443 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 9 19 theme catalytic 1463:1471 arg1 efficiency 1473:1482 the catalytic efficiency 1459:1482 the catalytic efficiency of the mutant 1459:1496 Furthermore, the catalytic efficiency of the mutant is reduced by approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin. 15809306 2 20 theme small 382:386 arg1 substrates 398:407 small dipeptide substrates 382:407 small dipeptide substrates 382:407 We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. 15809306 10 21 theme FAPalpha 1728:1735 arg1 specificity 1686:1696 substrate specificity 1676:1696 substrate specificity 1676:1696 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 10 21 theme FAPalpha 1728:1735 arg1 activity 1716:1723 endopeptidase activity 1702:1723 endopeptidase activity 1702:1723 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 6 22 theme enzyme 1029:1034 arg1 site 1017:1020 the active site 1006:1020 the active site of the enzyme 1006:1034 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 6 22 theme enzyme 1029:1034 arg1 enzyme 1029:1034 the enzyme 1025:1034 the enzyme 1025:1034 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 3 23 theme eight-bladed 521:532 arg1 domain 549:554 an eight-bladed beta-propeller domain 518:554 an eight-bladed beta-propeller domain 518:554 FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. 15809306 5 24 contain possesses 724:732 arg1 FAPalpha 699:706 FAPalpha 699:706 FAPalpha 699:706 Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. 15809306 5 24 contain possesses 724:732 arg1 DPPIV 717:721 DPPIV 717:721 DPPIV 717:721 Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. 15809306 5 24 contain possesses 724:732 arg2 activity 748:755 endopeptidase activity 734:755 endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides 734:812 Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. 15809306 5 25 theme N-terminal 764:773 arg1 peptides 805:812 N-terminal benzyloxycarbonyl (Z)-blocked peptides 764:812 N-terminal benzyloxycarbonyl (Z)-blocked peptides 764:812 Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. 15809306 2 26 theme high 307:310 arg1 structure 323:331 the first high resolution structure 297:331 the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates 297:407 We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. 15809306 1 27 theme epithelial 177:186 arg1 cancers 188:194 epithelial cancers 177:194 epithelial cancers 177:194 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 0 28 theme Structural 0:9 arg1 analysis 23:30 Structural and kinetic analysis 0:30 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 0:105 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 4 29 theme Known 557:561 arg1 dipeptides 569:578 Known DPPIV dipeptides 557:578 Known DPPIV dipeptides 557:578 Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. 15809306 6 30 theme structures 841:850 arg1 Comparison 815:824 Comparison 815:824 Comparison of the crystal structures of FAPalpha and DPPIV 815:872 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 9 31 theme Z-Gly-Pro-7-amino-4-methylcoumarin 1551:1584 arg1 cleavage 1539:1546 cleavage 1539:1546 cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin 1539:1584 Furthermore, the catalytic efficiency of the mutant is reduced by approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin. 15809306 2 32 theme first 301:305 arg1 structure 323:331 the first high resolution structure 297:331 the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates 297:407 We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. 15809306 2 33 theme kinetic 362:368 arg1 data 370:373 kinetic data 362:373 the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates 297:407 We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. 15809306 8 34 theme FAPalpha/A657D 1274:1287 arg1 analysis 1251:1258 kinetic analysis 1243:1258 kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), 1243:1402 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 7 35 dep as 1079:1080 arg1 in 1082:1083 in 1082:1083 in 1082:1083 Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. 15809306 0 36 theme kinetic 15:21 arg1 analysis 23:30 Structural and kinetic analysis 0:30 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 0:105 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 8 37 theme cat 1393:1395 arg1 /K 1397:1398 k(cat)/K 1391:1398 k(cat)/K(m) 1391:1401 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 8 37 theme cat 1393:1395 arg1 m 1400:1400 m 1400:1400 m 1400:1400 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 10 38 theme determinants 1643:1654 arg1 understanding 1612:1624 a clear understanding 1604:1624 a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha 1604:1735 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 6 39 theme major 887:891 arg1 difference 893:902 one major difference 883:902 one major difference 883:902 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 1 40 theme Fibroblast 107:116 arg1 alpha 137:141 Fibroblast activation protein alpha 107:141 Fibroblast activation protein alpha (FAPalpha) 107:152 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 1 40 theme Fibroblast 107:116 arg1 FAPalpha 144:151 FAPalpha 144:151 FAPalpha 144:151 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 4 41 theme DPPIV 563:567 arg1 dipeptides 569:578 Known DPPIV dipeptides 557:578 Known DPPIV dipeptides 557:578 Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. 15809306 10 42 theme molecular 1633:1641 arg1 determinants 1643:1654 the molecular determinants 1629:1654 the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha 1629:1735 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 4 43 from decrease 635:642 arg1 efficiency 657:666 catalytic efficiency 647:666 catalytic efficiency 647:666 Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. 15809306 10 44 theme endopeptidase 1702:1714 arg1 activity 1716:1723 endopeptidase activity 1702:1723 endopeptidase activity 1702:1723 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 3 45 theme dipeptidyl 430:439 arg1 peptidase 441:449 dipeptidyl peptidase IV 430:452 dipeptidyl peptidase IV (DPPIV) 430:460 FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. 15809306 3 45 theme dipeptidyl 430:439 arg1 DPPIV 455:459 DPPIV 455:459 DPPIV 455:459 FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. 15809306 5 46 theme -blocked 796:803 arg1 peptides 805:812 N-terminal benzyloxycarbonyl (Z)-blocked peptides 764:812 N-terminal benzyloxycarbonyl (Z)-blocked peptides 764:812 Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. 15809306 4 47 theme 100-fold 626:633 arg1 decrease 635:642 an approximately 100-fold decrease 609:642 an approximately 100-fold decrease in catalytic efficiency compared with DPPIV 609:686 Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. 15809306 2 48 theme dipeptide 388:396 arg1 substrates 398:407 small dipeptide substrates 382:407 small dipeptide substrates 382:407 We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. 15809306 0 49 theme specificity 49:59 arg1 analysis 23:30 Structural and kinetic analysis 0:30 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 0:105 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 6 50 theme DPPIV 868:872 arg1 structures 841:850 the crystal structures 829:850 the crystal structures of FAPalpha and DPPIV 829:872 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 9 51 theme mutant 1491:1496 arg1 efficiency 1473:1482 the catalytic efficiency 1459:1482 the catalytic efficiency of the mutant 1459:1496 Furthermore, the catalytic efficiency of the mutant is reduced by approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin. 15809306 1 52 theme activation 118:127 arg1 alpha 137:141 Fibroblast activation protein alpha 107:141 Fibroblast activation protein alpha (FAPalpha) 107:152 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 1 52 theme activation 118:127 arg1 FAPalpha 144:151 FAPalpha 144:151 FAPalpha 144:151 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 0 53 theme substrate 39:47 arg1 specificity 49:59 the substrate specificity 35:59 the substrate specificity of human fibroblast activation protein alpha 35:104 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 6 54 theme Glu 938:940 arg1 -Glu 946:949 Glu(203)-Glu(204) 938:954 Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV 938:996 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 7 55 theme lower 1163:1167 arg1 affinity 1169:1176 the lower affinity 1159:1176 the lower affinity for N-terminal amines by FAPalpha 1159:1210 Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. 15809306 6 56 dep -Glu 946:949 arg1 -Glu 978:981 Glu(205)-Glu(206) 970:986 Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV 938:996 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 8 57 theme kinetic 1243:1249 arg1 analysis 1251:1258 kinetic analysis 1243:1258 kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), 1243:1402 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 1 58 theme protein 129:135 arg1 alpha 137:141 Fibroblast activation protein alpha 107:141 Fibroblast activation protein alpha (FAPalpha) 107:152 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 1 58 theme protein 129:135 arg1 FAPalpha 144:151 FAPalpha 144:151 FAPalpha 144:151 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 7 59 from Ala 1037:1039 arg1 FAPalpha 1049:1056 FAPalpha 1049:1056 FAPalpha 1049:1056 Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. 15809306 6 60 theme FAPalpha 855:862 arg1 structures 841:850 the crystal structures 829:850 the crystal structures of FAPalpha and DPPIV 829:872 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 3 61 theme -like 461:465 arg1 fold 467:470 a dipeptidyl peptidase IV (DPPIV)-like fold 428:470 a dipeptidyl peptidase IV (DPPIV)-like fold 428:470 FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. 15809306 8 62 from increase 1338:1345 arg1 efficiency 1364:1373 the catalytic efficiency 1350:1373 the catalytic efficiency 1350:1373 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 8 63 theme 60-fold 1330:1336 arg1 increase 1338:1345 an approximately 60-fold increase 1313:1345 an approximately 60-fold increase in the catalytic efficiency 1313:1373 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 2 64 theme resolution 312:321 arg1 structure 323:331 the first high resolution structure 297:331 the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates 297:407 We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. 15809306 1 65 theme extracellular 223:235 arg1 remodeling 244:253 extracellular matrix remodeling 223:253 extracellular matrix remodeling 223:253 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 0 66 theme human 64:68 arg1 alpha 100:104 human fibroblast activation protein alpha 64:104 human fibroblast activation protein alpha 64:104 Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha. 15809306 4 67 theme catalytic 647:655 arg1 efficiency 657:666 catalytic efficiency 647:666 catalytic efficiency 647:666 Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. 15809306 7 68 theme N-terminal 1182:1191 arg1 amines 1193:1198 N-terminal amines 1182:1198 N-terminal amines 1182:1198 Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. 15809306 1 69 theme matrix 237:242 arg1 remodeling 244:253 extracellular matrix remodeling 223:253 extracellular matrix remodeling 223:253 Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. 15809306 8 70 theme substrates 1434:1443 arg1 cleavage 1412:1419 the cleavage 1408:1419 the cleavage of dipeptide substrates 1408:1443 This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. 15809306 6 71 theme motif 931:935 arg1 vicinity 911:918 the vicinity 907:918 the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme 907:1034 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 6 72 theme Glu 970:972 arg1 -Glu 978:981 Glu(205)-Glu(206) 970:986 Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV 938:996 Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. 15809306 10 73 theme substrate 1676:1684 arg1 specificity 1686:1696 substrate specificity 1676:1696 substrate specificity 1676:1696 Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha. 15809306 5 74 theme endopeptidase 734:746 arg1 activity 748:755 endopeptidase activity 734:755 endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides 734:812 Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. 17989695 3 0 theme factor 665:670 arg1 binding 679:685 vascular endothelial growth factor (VEGF) binding 637:685 vascular endothelial growth factor (VEGF) binding 637:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 1 1 theme system 252:257 arg1 development 225:235 the development 221:235 the development of the nervous system and the vasculature 221:277 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 17989695 3 2 with complex 569:575 arg1 antibodies 582:591 antibodies 582:591 antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding 582:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 3 3 theme vascular 637:644 arg1 VEGF 673:676 VEGF 673:676 VEGF 673:676 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 3 3 theme vascular 637:644 arg1 factor 665:670 vascular endothelial growth factor 637:670 vascular endothelial growth factor (VEGF) binding 637:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 4 4 theme a1 854:855 arg1 domain 857:862 the a1 domain 850:862 the a1 domain 850:862 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 3 5 theme endothelial 646:656 arg1 VEGF 673:676 VEGF 673:676 VEGF 673:676 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 3 5 theme endothelial 646:656 arg1 factor 665:670 vascular endothelial growth factor 637:670 vascular endothelial growth factor (VEGF) binding 637:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 3 6 theme crystal 510:516 arg1 structures 518:527 several crystal structures 502:527 several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding 502:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 6 7 theme possible 1072:1079 arg1 models 1081:1086 possible models 1072:1086 possible models for ligand binding to neuropilins 1072:1120 Based upon our structural and functional data, we propose possible models for ligand binding to neuropilins. 17989695 2 8 theme extracellular 284:296 arg1 portion 298:304 The extracellular portion 280:304 The extracellular portion of Nrp 280:311 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 1 9 theme vasculature 267:277 arg1 development 225:235 the development 221:235 the development of the nervous system and the vasculature 221:277 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 17989695 5 10 theme Nrp 1001:1003 arg1 binding 1005:1011 Nrp binding 1001:1011 Nrp binding 1001:1011 The locations of the antibody epitopes together with in vitro experiments indicate that VEGF and semaphorin do not directly compete for Nrp binding. 17989695 5 11 dep in 918:919 arg1 vitro 921:925 vitro 921:925 vitro 921:925 The locations of the antibody epitopes together with in vitro experiments indicate that VEGF and semaphorin do not directly compete for Nrp binding. 17989695 1 12 theme vascular 167:174 arg1 factors 195:201 vascular endothelial growth factors 167:201 vascular endothelial growth factors 167:201 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 17989695 2 13 theme receptor 465:472 arg1 dimerization 474:485 receptor dimerization 465:485 receptor dimerization (c) 465:489 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 2 13 theme receptor 465:472 arg1 c 488:488 c 488:488 c 488:488 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 4 14 theme b1 770:771 arg1 domains 781:787 the a2, b1, and b2 domains 762:787 the a2, b1, and b2 domains 762:787 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 6 15 theme functional 1044:1053 arg1 data 1055:1058 our structural and functional data 1025:1058 our structural and functional data 1025:1058 Based upon our structural and functional data, we propose possible models for ligand binding to neuropilins. 17989695 1 16 theme endothelial 176:186 arg1 factors 195:201 vascular endothelial growth factors 167:201 vascular endothelial growth factors 167:201 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 17989695 5 17 theme epitopes 895:902 arg1 locations 869:877 The locations 865:877 The locations of the antibody epitopes together with in vitro experiments 865:937 The locations of the antibody epitopes together with in vitro experiments indicate that VEGF and semaphorin do not directly compete for Nrp binding. 17989695 4 18 theme b2 778:779 arg1 domains 781:787 the a2, b1, and b2 domains 762:787 the a2, b1, and b2 domains 762:787 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 1 19 theme growth 188:193 arg1 factors 195:201 vascular endothelial growth factors 167:201 vascular endothelial growth factors 167:201 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 17989695 0 20 theme Structural 0:9 arg1 studies 11:17 Structural studies 0:17 Structural studies of neuropilin/antibody complexes 0:50 Structural studies of neuropilin/antibody complexes provide insights into semaphorin and VEGF binding. 17989695 0 21 theme neuropilin/antibody 22:40 arg1 complexes 42:50 neuropilin/antibody complexes 22:50 neuropilin/antibody complexes 22:50 Structural studies of neuropilin/antibody complexes provide insights into semaphorin and VEGF binding. 17989695 2 22 theme Nrp 309:311 arg1 portion 298:304 The extracellular portion 280:304 The extracellular portion of Nrp 280:311 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 5 23 theme in 918:919 arg1 experiments 927:937 in vitro experiments 918:937 in vitro experiments 918:937 The locations of the antibody epitopes together with in vitro experiments indicate that VEGF and semaphorin do not directly compete for Nrp binding. 17989695 2 24 theme necessary 402:410 arg1 domains 394:400 two domains 390:400 two domains necessary for VEGF binding (b1b2) 390:434 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 0 25 dep semaphorin 74:83 arg1 binding 94:100 binding 94:100 binding 94:100 Structural studies of neuropilin/antibody complexes provide insights into semaphorin and VEGF binding. 17989695 1 26 theme class 143:147 arg1 semaphorins 151:161 class 3 semaphorins 143:161 class 3 semaphorins 143:161 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 17989695 3 27 theme Nrp2 541:544 arg1 fragments 546:554 Nrp2 fragments 541:554 Nrp2 fragments 541:554 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 0 28 theme complexes 42:50 arg1 studies 11:17 Structural studies 0:17 Structural studies of neuropilin/antibody complexes 0:50 Structural studies of neuropilin/antibody complexes provide insights into semaphorin and VEGF binding. 17989695 2 29 theme VEGF 416:419 arg1 binding 421:427 VEGF binding 416:427 VEGF binding (b1b2) 416:434 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 2 29 theme VEGF 416:419 arg1 b1b2 430:433 b1b2 430:433 b1b2 430:433 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 3 30 theme Nrp1 532:535 arg1 structures 518:527 several crystal structures 502:527 several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding 502:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 6 31 theme structural 1029:1038 arg1 data 1055:1058 our structural and functional data 1025:1058 our structural and functional data 1025:1058 Based upon our structural and functional data, we propose possible models for ligand binding to neuropilins. 17989695 4 32 theme packed 804:809 arg1 core 811:814 a tightly packed core 794:814 a tightly packed core that is only loosely connected to the a1 domain 794:862 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 4 33 theme a2 766:767 arg1 domains 781:787 the a2, b1, and b2 domains 762:787 the a2, b1, and b2 domains 762:787 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 5 34 theme antibody 886:893 arg1 epitopes 895:902 the antibody epitopes 882:902 the antibody epitopes 882:902 The locations of the antibody epitopes together with in vitro experiments indicate that VEGF and semaphorin do not directly compete for Nrp binding. 17989695 4 35 theme domain 734:739 arg1 arrangement 741:751 an unexpected domain arrangement 720:751 an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain 720:862 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 3 36 theme fragments 546:554 arg1 structures 518:527 several crystal structures 502:527 several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding 502:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 3 37 theme several 502:508 arg1 structures 518:527 several crystal structures 502:527 several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding 502:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 4 38 theme unexpected 723:732 arg1 arrangement 741:751 an unexpected domain arrangement 720:751 an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain 720:862 In these structures, Nrps adopt an unexpected domain arrangement in which the a2, b1, and b2 domains form a tightly packed core that is only loosely connected to the a1 domain. 17989695 6 39 theme ligand 1092:1097 arg1 binding 1099:1105 ligand binding 1092:1105 ligand binding to neuropilins 1092:1120 Based upon our structural and functional data, we propose possible models for ligand binding to neuropilins. 17989695 2 40 theme semaphorin 363:372 arg1 a1a2 383:386 a1a2 383:386 a1a2 383:386 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 2 40 theme semaphorin 363:372 arg1 binding 374:380 semaphorin binding 363:380 semaphorin binding (a1a2) 363:387 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 2 41 theme critical 452:459 arg1 domain 445:450 one domain 441:450 one domain critical for receptor dimerization (c) 441:489 The extracellular portion of Nrp is composed of two domains that are essential for semaphorin binding (a1a2), two domains necessary for VEGF binding (b1b2), and one domain critical for receptor dimerization (c). 17989695 3 42 theme growth 658:663 arg1 VEGF 673:676 VEGF 673:676 VEGF 673:676 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 3 42 theme growth 658:663 arg1 factor 665:670 vascular endothelial growth factor 637:670 vascular endothelial growth factor (VEGF) binding 637:685 We report several crystal structures of Nrp1 and Nrp2 fragments alone and in complex with antibodies that selectively block either semaphorin or vascular endothelial growth factor (VEGF) binding. 17989695 1 43 theme nervous 244:250 arg1 system 252:257 the nervous system 240:257 the nervous system 240:257 Neuropilins (Nrps) are co-receptors for class 3 semaphorins and vascular endothelial growth factors and important for the development of the nervous system and the vasculature. 30312582 0 0 theme Cell 56:59 arg1 Shape 61:65 Human Cell Shape 50:65 Human Cell Shape 50:65 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 5 1 theme stress 723:728 arg1 fibers 730:735 actin stress fibers 717:735 actin stress fibers 717:735 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 0 2 theme Human 50:54 arg1 Shape 61:65 Human Cell Shape 50:65 Human Cell Shape 50:65 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 9 3 theme decarboxylase 1269:1281 arg1 antizyme-1 1283:1292 ornithine decarboxylase antizyme-1 1259:1292 ornithine decarboxylase antizyme-1 1259:1292 Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. 30312582 6 4 theme endogenous 788:797 arg1 APCN 799:802 endogenous APCN 788:802 endogenous APCN 788:802 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 11 5 theme actin 1468:1472 arg1 dynamics 1452:1459 the dynamics 1448:1459 the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells 1448:1559 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 4 6 theme placental 476:484 arg1 trophoblasts 486:497 placental trophoblasts 476:497 placental trophoblasts 476:497 APCN is physiologically expressed in placental trophoblasts, skeletal and hearth muscle, and kidney and pancreas. 30312582 5 7 theme APCN 571:574 arg1 Overexpression 553:566 Overexpression 553:566 Overexpression of APCN (cDNA) in various cell lines 553:603 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 11 8 theme cytoskeletal 1474:1485 arg1 actin 1468:1472 the actin cytoskeletal 1464:1485 the actin cytoskeletal 1464:1485 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 0 9 theme Shape 61:65 arg1 Regulator 37:45 A Regulator 35:45 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 0:86 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 10 10 theme APCN 1406:1409 arg1 description 1391:1401 the first functional description 1370:1401 the first functional description of APCN 1370:1409 This is the first functional description of APCN. 30312582 10 10 theme APCN 1406:1409 arg1 This 1362:1365 This 1362:1365 This 1362:1365 This is the first functional description of APCN. 30312582 6 11 theme cancers 776:782 arg1 staining 758:765 Immunohistochemical staining 738:765 Immunohistochemical staining of human cancers for endogenous APCN 738:802 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 11 12 theme invasive 1520:1527 arg1 potential 1536:1544 invasive growth potential 1520:1544 invasive growth potential 1520:1544 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 0 13 theme Growth 80:85 arg1 Regulator 37:45 A Regulator 35:45 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 0:86 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 6 14 theme intratumoral 869:880 arg1 cells 882:886 intratumoral cells 869:886 intratumoral cells 869:886 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 6 15 theme human 770:774 arg1 cancers 776:782 human cancers 770:782 human cancers 770:782 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 0 16 theme Invasive 71:78 arg1 Growth 80:85 Invasive Growth 71:85 Invasive Growth 71:85 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 3 17 dep intracytoplasmic 350:365 arg1 whereas 368:374 whereas 368:374 whereas 368:374 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 11 18 theme tumor 1549:1553 arg1 cells 1555:1559 tumor cells 1549:1559 tumor cells 1549:1559 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 8 19 theme carboxyterminal 1061:1075 arg1 essential 1119:1127 essential 1119:1127 essential 1119:1127 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 8 19 theme carboxyterminal 1061:1075 arg1 stretch 1077:1083 A conserved carboxyterminal stretch 1049:1083 A conserved carboxyterminal stretch of 21 amino acids 1049:1101 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 7 20 from ability 962:968 arg1 medium 1002:1007 semisolid medium 992:1007 semisolid medium 992:1007 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 3 21 theme conserved 329:337 arg1 protein 339:345 the evolutionarily conserved protein 310:345 the evolutionarily conserved protein 310:345 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 5 22 theme fibers 730:735 arg1 disappearance 700:712 disappearance 700:712 disappearance of actin stress fibers 700:735 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 5 23 theme various 586:592 arg1 lines 599:603 various cell lines 586:603 various cell lines 586:603 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 11 24 theme growth 1529:1534 arg1 potential 1536:1544 invasive growth potential 1520:1544 invasive growth potential 1520:1544 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 2 25 theme protein 267:273 arg1 APCN 289:292 APCN 289:292 APCN 289:292 Because of its abundant expression in astrocytes, it was called the protein astroprincin (APCN). 30312582 2 25 theme protein 267:273 arg1 astroprincin 275:286 the protein astroprincin 263:286 the protein astroprincin (APCN) 263:293 Because of its abundant expression in astrocytes, it was called the protein astroprincin (APCN). 30312582 1 26 theme 98-kDa 137:142 arg1 glycoprotein 165:176 a 98-kDa type 1 transmembrane glycoprotein 135:176 a 98-kDa type 1 transmembrane glycoprotein of unknown function 135:196 Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. 30312582 3 27 theme protein 339:345 arg1 thirds 300:305 Two thirds 296:305 Two thirds of the evolutionarily conserved protein 296:345 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 10 28 theme first 1374:1378 arg1 description 1391:1401 the first functional description 1370:1401 the first functional description of APCN 1370:1409 This is the first functional description of APCN. 30312582 10 28 theme first 1374:1378 arg1 This 1362:1365 This 1362:1365 This 1362:1365 This is the first functional description of APCN. 30312582 1 29 gly glycoprotein 165:176 arg1 glycoprotein 165:176 a 98-kDa type 1 transmembrane glycoprotein 135:176 a 98-kDa type 1 transmembrane glycoprotein of unknown function 135:196 Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. 30312582 6 30 theme Immunohistochemical 738:756 arg1 staining 758:765 Immunohistochemical staining 738:765 Immunohistochemical staining of human cancers for endogenous APCN 738:802 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 11 31 theme shape 1510:1514 arg1 dynamics 1452:1459 the dynamics 1448:1459 the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells 1448:1559 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 3 32 theme extracellular 380:392 arg1 domain 394:399 the extracellular domain 376:399 the extracellular domain 376:399 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 10 33 theme functional 1380:1389 arg1 description 1391:1401 the first functional description 1370:1401 the first functional description of APCN 1370:1409 This is the first functional description of APCN. 30312582 10 33 theme functional 1380:1389 arg1 This 1362:1365 This 1362:1365 This 1362:1365 This is the first functional description of APCN. 30312582 7 34 theme control 1034:1040 arg1 cells 1042:1046 control cells 1034:1046 control cells 1034:1046 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 7 35 theme APCN 939:942 arg1 cDNA 944:947 APCN cDNA 939:947 APCN cDNA 939:947 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 7 36 dep acquired 949:956 arg1 seen 1024:1027 seen 1024:1027 acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells 949:1046 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 3 37 theme side 426:429 arg1 chains 431:436 two N-glycosidic side chains 409:436 two N-glycosidic side chains 409:436 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 9 38 theme two-hybrid 1190:1199 arg1 screening 1201:1209 Yeast two-hybrid screening 1184:1209 Yeast two-hybrid screening 1184:1209 Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. 30312582 2 39 from expression 223:232 arg1 astrocytes 237:246 astrocytes 237:246 astrocytes 237:246 Because of its abundant expression in astrocytes, it was called the protein astroprincin (APCN). 30312582 1 40 theme transmembrane 151:163 arg1 glycoprotein 165:176 a 98-kDa type 1 transmembrane glycoprotein 135:176 a 98-kDa type 1 transmembrane glycoprotein of unknown function 135:196 Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. 30312582 5 41 from Overexpression 553:566 arg1 lines 599:603 various cell lines 586:603 various cell lines 586:603 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 5 42 theme APCN 668:671 arg1 expression 673:682 APCN expression 668:682 APCN expression 668:682 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 6 43 theme tumor 843:847 arg1 cells 849:853 invasive tumor cells 834:853 invasive tumor cells 834:853 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 9 44 theme ornithine 1259:1267 arg1 decarboxylase 1269:1281 ornithine decarboxylase 1259:1281 ornithine decarboxylase antizyme-1 1259:1292 Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. 30312582 1 45 theme type 144:147 arg1 glycoprotein 165:176 a 98-kDa type 1 transmembrane glycoprotein 135:176 a 98-kDa type 1 transmembrane glycoprotein of unknown function 135:196 Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. 30312582 8 46 theme invasive 1167:1174 arg1 growth 1176:1181 invasive growth 1167:1181 invasive growth 1167:1181 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 5 47 theme expression 673:682 arg1 knockdown 655:663 knockdown 655:663 knockdown of APCN expression by siRNA 655:691 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 6 48 theme invasive 834:841 arg1 cells 849:853 invasive tumor cells 834:853 invasive tumor cells 834:853 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 7 49 theme semisolid 992:1000 arg1 medium 1002:1007 semisolid medium 992:1007 semisolid medium 992:1007 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 3 50 theme N-glycosidic 413:424 arg1 chains 431:436 two N-glycosidic side chains 409:436 two N-glycosidic side chains 409:436 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 6 51 from expression 820:829 arg1 cells 849:853 invasive tumor cells 834:853 invasive tumor cells 834:853 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 5 52 theme slender 626:632 arg1 projections 634:644 slender projections 626:644 slender projections 626:644 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 9 53 theme several 1220:1226 arg1 partners 1240:1247 several interactive partners 1220:1247 several interactive partners 1220:1247 Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. 30312582 8 54 dep cell 1148:1151 arg1 sprouting 1153:1161 sprouting 1153:1161 sprouting 1153:1161 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 8 55 theme conserved 1051:1059 arg1 essential 1119:1127 essential 1119:1127 essential 1119:1127 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 8 55 theme conserved 1051:1059 arg1 stretch 1077:1083 A conserved carboxyterminal stretch 1049:1083 A conserved carboxyterminal stretch of 21 amino acids 1049:1101 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 4 56 theme hearth 513:518 arg1 muscle 520:525 hearth muscle 513:525 hearth muscle 513:525 APCN is physiologically expressed in placental trophoblasts, skeletal and hearth muscle, and kidney and pancreas. 30312582 5 57 theme projections 634:644 arg1 sprouting 613:621 sprouting 613:621 sprouting of slender projections 613:644 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 9 58 theme interactive 1228:1238 arg1 partners 1240:1247 several interactive partners 1220:1247 several interactive partners 1220:1247 Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. 30312582 7 59 theme growth 982:987 arg1 ability 962:968 the ability 958:968 the ability of invasive growth in semisolid medium 958:1007 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 5 60 theme cell 594:597 arg1 lines 599:603 various cell lines 586:603 various cell lines 586:603 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 7 61 theme melanoma 895:902 arg1 SK-MEL-28 911:919 SK-MEL-28 911:919 SK-MEL-28 911:919 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 7 61 theme melanoma 895:902 arg1 cells 904:908 Human melanoma cells 889:908 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA 889:947 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 9 62 theme Yeast 1184:1188 arg1 screening 1201:1209 Yeast two-hybrid screening 1184:1209 Yeast two-hybrid screening 1184:1209 Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. 30312582 8 63 theme acids 1097:1101 arg1 essential 1119:1127 essential 1119:1127 essential 1119:1127 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 8 63 theme acids 1097:1101 arg1 stretch 1077:1083 A conserved carboxyterminal stretch 1049:1083 A conserved carboxyterminal stretch of 21 amino acids 1049:1101 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 6 64 theme elevated 811:818 arg1 expression 820:829 elevated expression 811:829 elevated expression in invasive tumor cells 811:853 Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. 30312582 11 65 theme cell 1505:1508 arg1 shape 1510:1514 cell shape 1505:1514 cell shape 1505:1514 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 1 66 theme unknown 181:187 arg1 function 189:196 unknown function 181:196 unknown function 181:196 Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. 30312582 8 67 theme 21 amino 1088:1095 arg1 acids 1097:1101 21 amino acids 1088:1101 21 amino acids 1088:1101 A conserved carboxyterminal stretch of 21 amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. 30312582 3 68 contain carries 401:407 arg1 domain 394:399 the extracellular domain 376:399 the extracellular domain 376:399 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 3 68 contain carries 401:407 arg2 chains 431:436 two N-glycosidic side chains 409:436 two N-glycosidic side chains 409:436 Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. 30312582 2 69 theme abundant 214:221 arg1 expression 223:232 its abundant expression 210:232 its abundant expression in astrocytes 210:246 Because of its abundant expression in astrocytes, it was called the protein astroprincin (APCN). 30312582 1 70 theme function 189:196 arg1 glycoprotein 165:176 a 98-kDa type 1 transmembrane glycoprotein 135:176 a 98-kDa type 1 transmembrane glycoprotein of unknown function 135:196 Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. 30312582 7 71 theme invasive 973:980 arg1 growth 982:987 invasive growth 973:987 invasive growth 973:987 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 7 72 theme Human 889:893 arg1 SK-MEL-28 911:919 SK-MEL-28 911:919 SK-MEL-28 911:919 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 7 72 theme Human 889:893 arg1 cells 904:908 Human melanoma cells 889:908 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA 889:947 Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. 30312582 5 73 theme actin 717:721 arg1 fibers 730:735 actin stress fibers 717:735 actin stress fibers 717:735 Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. 30312582 11 74 theme cells 1555:1559 arg1 potential 1536:1544 invasive growth potential 1520:1544 invasive growth potential 1520:1544 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 11 74 theme cells 1555:1559 arg1 shape 1510:1514 cell shape 1505:1514 cell shape 1505:1514 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 11 74 theme cells 1555:1559 arg1 actin 1468:1472 the actin cytoskeletal 1464:1485 the actin cytoskeletal 1464:1485 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 30312582 0 75 dep Astroprincin 0:11 arg1 FAM171A1 14:21 FAM171A1 14:21 FAM171A1 14:21 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 0 75 dep Astroprincin 0:11 arg1 Regulator 37:45 A Regulator 35:45 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 0:86 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 0 75 dep Astroprincin 0:11 arg1 C10orf38 24:31 C10orf38 24:31 C10orf38 24:31 Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth. 30312582 11 76 dep shape 1510:1514 arg1 the 1501:1503 the 1501:1503 the 1501:1503 These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells. 24226769 6 0 with patient 780:786 arg1 heterotaxy 793:802 heterotaxy 793:802 heterotaxy 793:802 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 8 1 theme Xenopus 1182:1188 arg1 cilia 1211:1215 Xenopus left-right organizer cilia 1182:1215 Xenopus left-right organizer cilia 1182:1215 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 5 2 theme enzyme 665:670 arg1 GALNT11 672:678 the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 618:678 the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 618:678 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 5 2 theme enzyme 665:670 arg1 crucial 683:689 crucial 683:689 crucial 683:689 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 8 3 theme quantitative 1142:1153 arg1 technique 1168:1176 a quantitative live imaging technique 1140:1176 a quantitative live imaging technique for Xenopus left-right organizer cilia 1140:1215 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 7 4 theme human 913:917 arg1 peptides 926:933 human NOTCH1 peptides 913:933 human NOTCH1 peptides 913:933 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 9 5 theme cilia 1460:1464 arg1 expense 1440:1446 the expense 1436:1446 the expense of immotile cilia 1436:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 7 6 theme repeats 1110:1116 arg1 modification 1081:1092 modification 1081:1092 modification of specific EGF repeats 1081:1116 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 7 7 theme Notch 979:983 arg1 activation 985:994 Notch activation 979:994 Notch activation 979:994 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 1 8 theme laterality 148:157 arg1 Heterotaxy 89:98 Heterotaxy 89:98 Heterotaxy 89:98 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 1 8 theme laterality 148:157 arg1 disorder 105:112 a disorder 103:112 a disorder 103:112 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 11 9 from organizer 1801:1809 arg1 cilia 1777:1781 motile and immotile cilia 1757:1781 motile and immotile cilia at the left-right organizer to determine laterality 1757:1833 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 4 10 theme cilia 571:575 arg1 types 577:581 these two cilia types 561:581 these two cilia types 561:581 The mechanism that specifies these two cilia types remains unknown. 24226769 8 11 theme organizer 1201:1209 arg1 cilia 1211:1215 Xenopus left-right organizer cilia 1182:1215 Xenopus left-right organizer cilia 1182:1215 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 7 12 theme activation 985:994 arg1 mechanism 966:974 a mechanism 964:974 a mechanism of Notch activation 964:994 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 2 13 theme heterotaxy 276:285 arg1 cases 261:265 most cases 256:265 most cases of human heterotaxy 256:285 The aetiology and mechanisms underlying most cases of human heterotaxy are poorly understood. 24226769 8 14 theme left-right 1190:1199 arg1 cilia 1211:1215 Xenopus left-right organizer cilia 1182:1215 Xenopus left-right organizer cilia 1182:1215 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 11 15 theme essential 1731:1739 arg1 balance 1741:1747 an essential balance 1728:1747 an essential balance between motile and immotile cilia at the left-right organizer to determine laterality 1728:1833 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 2 16 theme human 270:274 arg1 heterotaxy 276:285 human heterotaxy 270:285 human heterotaxy 270:285 The aetiology and mechanisms underlying most cases of human heterotaxy are poorly understood. 24226769 8 17 theme left-right 1347:1356 arg1 organizer 1358:1366 the left-right organizer 1343:1366 the left-right organizer 1343:1366 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 3 18 theme sensory 460:466 arg1 cilia 468:472 immotile sensory cilia 451:472 immotile sensory cilia 451:472 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 8 19 theme Galnt11-mediated 1231:1246 arg1 signalling 1255:1264 Galnt11-mediated Notch1 signalling 1231:1264 Galnt11-mediated Notch1 signalling 1231:1264 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 9 20 theme laterality 1481:1490 arg1 defect 1492:1497 a laterality defect 1479:1497 a laterality defect reminiscent of loss of the ciliary sensor Pkd2 1479:1544 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 9 21 theme loss 1514:1517 arg1 reminiscent 1499:1509 reminiscent 1499:1509 reminiscent 1499:1509 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 9 22 from ratio 1411:1415 arg1 expense 1440:1446 the expense 1436:1446 the expense of immotile cilia 1436:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 7 23 theme ADAM17-mediated 1017:1031 arg1 shedding 1044:1051 ADAM17-mediated ectodomain shedding 1017:1051 ADAM17-mediated ectodomain shedding of the Notch receptor 1017:1073 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 0 24 theme heterotaxy 4:13 arg1 GALNT11 20:26 The heterotaxy gene GALNT11 0:26 The heterotaxy gene GALNT11 0:26 The heterotaxy gene GALNT11 glycosylates Notch to orchestrate cilia type and laterality. 24226769 9 25 theme notch1 1380:1385 arg1 depletion 1387:1395 notch1 depletion 1380:1395 notch1 depletion 1380:1395 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 5 26 theme N-acetylgalactosamine-type 622:647 arg1 GALNT11 672:678 the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 618:678 the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 618:678 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 5 26 theme N-acetylgalactosamine-type 622:647 arg1 crucial 683:689 crucial 683:689 crucial 683:689 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 10 27 theme Notch 1560:1564 arg1 overexpression 1566:1579 Notch overexpression 1560:1579 Notch overexpression 1560:1579 By contrast, Notch overexpression decreases this ratio, mimicking the ciliopathy primary ciliary dyskinesia. 24226769 3 28 theme leftward 417:424 arg1 flow 426:429 leftward flow 417:429 leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals 417:529 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 5 29 theme O-glycosylation 649:663 arg1 GALNT11 672:678 the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 618:678 the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 618:678 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 5 29 theme O-glycosylation 649:663 arg1 crucial 683:689 crucial 683:689 crucial 683:689 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 8 30 theme motile 1314:1319 arg1 cilia 1334:1338 motile and immotile cilia 1314:1338 motile and immotile cilia 1314:1338 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 7 31 gly O-glycosylates 898:911 arg1 peptides 926:933 human NOTCH1 peptides 913:933 human NOTCH1 peptides 913:933 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 3 32 theme embryonic 357:365 arg1 organizer 378:386 the embryonic left-right organizer 353:386 the embryonic left-right organizer 353:386 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 11 33 theme human 1869:1873 arg1 heterotaxy 1875:1884 human heterotaxy 1869:1884 human heterotaxy 1869:1884 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 7 34 theme specific 1097:1104 arg1 repeats 1110:1116 specific EGF repeats 1097:1116 specific EGF repeats 1097:1116 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 8 35 from organizer 1358:1366 arg1 ratio 1305:1309 ratio 1305:1309 ratio 1305:1309 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 8 35 from organizer 1358:1366 arg1 distribution 1288:1299 spatial distribution 1280:1299 spatial distribution 1280:1299 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 3 36 theme immotile 451:458 arg1 cilia 468:472 immotile sensory cilia 451:472 immotile sensory cilia 451:472 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 3 37 theme left-right 367:376 arg1 organizer 378:386 the embryonic left-right organizer 353:386 the embryonic left-right organizer 353:386 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 11 38 theme novel 1849:1853 arg1 mechanism 1855:1863 a novel mechanism 1847:1863 a novel mechanism for human heterotaxy 1847:1884 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 7 39 theme EGF 1106:1108 arg1 repeats 1110:1116 specific EGF repeats 1097:1116 specific EGF repeats 1097:1116 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 8 40 dep distribution 1288:1299 arg1 the 1276:1278 the 1276:1278 the 1276:1278 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 1 41 theme major 184:188 arg1 disease 207:213 major congenital heart disease 184:213 major congenital heart disease 184:213 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 1 42 theme patterning 133:142 arg1 Heterotaxy 89:98 Heterotaxy 89:98 Heterotaxy 89:98 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 1 42 theme patterning 133:142 arg1 disorder 105:112 a disorder 103:112 a disorder 103:112 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 9 43 from expense 1440:1446 arg1 ratio 1411:1415 the ratio 1407:1415 the ratio of motile cilia at the expense of immotile cilia 1407:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 9 43 from expense 1440:1446 arg1 cilia 1427:1431 motile cilia 1420:1431 motile cilia at the expense of immotile cilia 1420:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 6 44 theme Notch 872:876 arg1 signalling 878:887 Notch signalling 872:887 Notch signalling 872:887 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 11 45 theme immotile 1768:1775 arg1 cilia 1777:1781 motile and immotile cilia 1757:1781 motile and immotile cilia at the left-right organizer to determine laterality 1757:1833 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 1 46 theme congenital 190:199 arg1 disease 207:213 major congenital heart disease 184:213 major congenital heart disease 184:213 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 10 47 theme ciliary 1636:1642 arg1 dyskinesia 1644:1653 the ciliopathy primary ciliary dyskinesia 1613:1653 the ciliopathy primary ciliary dyskinesia 1613:1653 By contrast, Notch overexpression decreases this ratio, mimicking the ciliopathy primary ciliary dyskinesia. 24226769 10 48 theme ciliopathy 1617:1626 arg1 dyskinesia 1644:1653 the ciliopathy primary ciliary dyskinesia 1613:1653 the ciliopathy primary ciliary dyskinesia 1613:1653 By contrast, Notch overexpression decreases this ratio, mimicking the ciliopathy primary ciliary dyskinesia. 24226769 8 49 theme cilia 1334:1338 arg1 ratio 1305:1309 ratio 1305:1309 ratio 1305:1309 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 8 49 theme cilia 1334:1338 arg1 distribution 1288:1299 spatial distribution 1280:1299 spatial distribution 1280:1299 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 8 50 theme Notch1 1248:1253 arg1 signalling 1255:1264 Galnt11-mediated Notch1 signalling 1231:1264 Galnt11-mediated Notch1 signalling 1231:1264 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 1 51 theme heart 201:205 arg1 disease 207:213 major congenital heart disease 184:213 major congenital heart disease 184:213 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 8 52 theme spatial 1280:1286 arg1 distribution 1288:1299 spatial distribution 1280:1299 spatial distribution 1280:1299 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 9 53 theme motile 1420:1425 arg1 cilia 1427:1431 motile cilia 1420:1431 motile cilia at the expense of immotile cilia 1420:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 10 54 theme primary 1628:1634 arg1 dyskinesia 1644:1653 the ciliopathy primary ciliary dyskinesia 1613:1653 the ciliopathy primary ciliary dyskinesia 1613:1653 By contrast, Notch overexpression decreases this ratio, mimicking the ciliopathy primary ciliary dyskinesia. 24226769 8 55 theme imaging 1160:1166 arg1 technique 1168:1176 a quantitative live imaging technique 1140:1176 a quantitative live imaging technique for Xenopus left-right organizer cilia 1140:1215 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 8 56 theme live 1155:1158 arg1 technique 1168:1176 a quantitative live imaging technique 1140:1176 a quantitative live imaging technique for Xenopus left-right organizer cilia 1140:1215 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 9 57 theme sensor 1534:1539 arg1 Pkd2 1541:1544 the ciliary sensor Pkd2 1522:1544 the ciliary sensor Pkd2 1522:1544 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 3 58 theme motile 395:400 arg1 cilia 402:406 motile cilia 395:406 motile cilia 395:406 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 9 59 theme cilia 1427:1431 arg1 ratio 1411:1415 the ratio 1407:1415 the ratio of motile cilia at the expense of immotile cilia 1407:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 5 60 theme such 694:697 arg1 determination 699:711 such determination 694:711 such determination 694:711 Here we show that the N-acetylgalactosamine-type O-glycosylation enzyme GALNT11 is crucial to such determination. 24226769 11 61 theme left-right 1790:1799 arg1 organizer 1801:1809 the left-right organizer 1786:1809 the left-right organizer to determine laterality 1786:1833 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 7 62 theme ectodomain 1033:1042 arg1 shedding 1044:1051 ADAM17-mediated ectodomain shedding 1017:1051 ADAM17-mediated ectodomain shedding of the Notch receptor 1017:1073 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 9 63 theme Pkd2 1541:1544 arg1 loss 1514:1517 loss 1514:1517 loss of the ciliary sensor Pkd2 1514:1544 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 7 64 theme Notch 1060:1064 arg1 receptor 1066:1073 the Notch receptor 1056:1073 the Notch receptor 1056:1073 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 0 65 gly glycosylates 28:39 arg1 Notch 41:45 Notch 41:45 Notch 41:45 The heterotaxy gene GALNT11 glycosylates Notch to orchestrate cilia type and laterality. 24226769 2 66 theme most 256:259 arg1 cases 261:265 most cases 256:265 most cases of human heterotaxy 256:285 The aetiology and mechanisms underlying most cases of human heterotaxy are poorly understood. 24226769 1 67 theme left-right 117:126 arg1 patterning 133:142 left-right body patterning 117:142 left-right body patterning 117:142 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 8 68 theme immotile 1325:1332 arg1 cilia 1334:1338 motile and immotile cilia 1314:1338 motile and immotile cilia 1314:1338 We further developed a quantitative live imaging technique for Xenopus left-right organizer cilia and show that Galnt11-mediated Notch1 signalling modulates the spatial distribution and ratio of motile and immotile cilia at the left-right organizer. 24226769 7 69 theme receptor 1066:1073 arg1 shedding 1044:1051 ADAM17-mediated ectodomain shedding 1017:1051 ADAM17-mediated ectodomain shedding of the Notch receptor 1017:1073 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 6 70 theme disease 762:768 arg1 gene 770:773 a candidate disease gene 750:773 a candidate disease gene 750:773 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 6 70 theme disease 762:768 arg1 GALNT11 739:745 GALNT11 739:745 GALNT11 739:745 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 1 71 theme body 128:131 arg1 patterning 133:142 left-right body patterning 117:142 left-right body patterning 117:142 Heterotaxy is a disorder of left-right body patterning, or laterality, that is associated with major congenital heart disease. 24226769 7 72 theme NOTCH1 919:924 arg1 peptides 926:933 human NOTCH1 peptides 913:933 human NOTCH1 peptides 913:933 GALNT11 O-glycosylates human NOTCH1 peptides in vitro, thereby supporting a mechanism of Notch activation either by increasing ADAM17-mediated ectodomain shedding of the Notch receptor or by modification of specific EGF repeats. 24226769 9 73 theme ciliary 1526:1532 arg1 Pkd2 1541:1544 the ciliary sensor Pkd2 1522:1544 the ciliary sensor Pkd2 1522:1544 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 6 74 theme Xenopus 829:835 arg1 tropicalis 837:846 Xenopus tropicalis 829:846 Xenopus tropicalis 829:846 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 6 75 theme candidate 752:760 arg1 gene 770:773 a candidate disease gene 750:773 a candidate disease gene 750:773 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 6 75 theme candidate 752:760 arg1 GALNT11 739:745 GALNT11 739:745 GALNT11 739:745 We previously identified GALNT11 as a candidate disease gene in a patient with heterotaxy, and now demonstrate, in Xenopus tropicalis, that galnt11 activates Notch signalling. 24226769 3 76 theme downstream 501:510 arg1 signals 523:529 downstream asymmetric signals 501:529 downstream asymmetric signals 501:529 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 0 77 theme cilia 62:66 arg1 type 68:71 cilia type 62:71 cilia type 62:71 The heterotaxy gene GALNT11 glycosylates Notch to orchestrate cilia type and laterality. 24226769 9 78 theme reminiscent 1499:1509 arg1 defect 1492:1497 a laterality defect 1479:1497 a laterality defect reminiscent of loss of the ciliary sensor Pkd2 1479:1544 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24226769 0 79 theme gene 15:18 arg1 GALNT11 20:26 The heterotaxy gene GALNT11 0:26 The heterotaxy gene GALNT11 0:26 The heterotaxy gene GALNT11 glycosylates Notch to orchestrate cilia type and laterality. 24226769 3 80 theme asymmetric 512:521 arg1 signals 523:529 downstream asymmetric signals 501:529 downstream asymmetric signals 501:529 In vertebrates, laterality is initiated at the embryonic left-right organizer, where motile cilia generate leftward flow that is detected by immotile sensory cilia, which transduce flow into downstream asymmetric signals. 24226769 11 81 theme motile 1757:1762 arg1 cilia 1777:1781 motile and immotile cilia 1757:1781 motile and immotile cilia at the left-right organizer to determine laterality 1757:1833 Together our data demonstrate that Galnt11 modifies Notch, establishing an essential balance between motile and immotile cilia at the left-right organizer to determine laterality, and reveal a novel mechanism for human heterotaxy. 24226769 9 82 theme immotile 1451:1458 arg1 cilia 1460:1464 immotile cilia 1451:1464 immotile cilia 1451:1464 galnt11 or notch1 depletion increases the ratio of motile cilia at the expense of immotile cilia and produces a laterality defect reminiscent of loss of the ciliary sensor Pkd2. 24927598 0 0 theme metal 102:106 arg1 transporter 108:118 the plasma membrane metal transporter ZIP14 82:124 the plasma membrane metal transporter ZIP14 82:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 8 1 theme proteins 1231:1238 arg1 degradation 1188:1198 the degradation 1184:1198 the degradation of endocytosed plasma membrane proteins 1184:1238 These findings highlight the importance of proteasomes in the degradation of endocytosed plasma membrane proteins. 24927598 0 2 theme membrane 93:100 arg1 transporter 108:118 the plasma membrane metal transporter ZIP14 82:124 the plasma membrane metal transporter ZIP14 82:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 6 3 theme ZIP14 917:921 arg1 levels 907:912 higher steady-state levels 887:912 higher steady-state levels of ZIP14 887:921 Iron inhibited membrane extraction of internalized ZIP14, resulting in higher steady-state levels of ZIP14. 24927598 7 4 theme N-linked 943:950 arg1 glycosylation 953:965 Asparagine-linked (N-linked) glycosylation 924:965 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, 924:1015 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 5 5 theme endoplasmic 754:764 arg1 pathway 807:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 7 6 theme efficient 1034:1042 arg1 extraction 1053:1062 efficient membrane extraction 1034:1062 efficient membrane extraction of ZIP14 1034:1071 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 5 7 theme reticulum-associated 766:785 arg1 pathway 807:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 5 8 theme endoplasmic 690:700 arg1 reticulum 702:710 the endoplasmic reticulum 686:710 the endoplasmic reticulum 686:710 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 7 9 theme Asparagine-linked 924:940 arg1 glycosylation 953:965 Asparagine-linked (N-linked) glycosylation 924:965 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, 924:1015 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 3 10 theme identified 369:378 arg1 ZIP14 352:356 ZIP14 352:356 ZIP14 352:356 ZIP14 is a newly identified iron transporter with multitransmembrane domains. 24927598 3 10 theme identified 369:378 arg1 transporter 385:395 a newly identified iron transporter 361:395 a newly identified iron transporter with multitransmembrane domains 361:427 ZIP14 is a newly identified iron transporter with multitransmembrane domains. 24927598 6 11 theme ZIP14 867:871 arg1 extraction 840:849 membrane extraction 831:849 membrane extraction of internalized ZIP14 831:871 Iron inhibited membrane extraction of internalized ZIP14, resulting in higher steady-state levels of ZIP14. 24927598 5 12 theme degradation 795:805 arg1 pathway 807:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 7 13 link Asparagine-linked 924:940 arg1 glycosylation 953:965 Asparagine-linked (N-linked) glycosylation 924:965 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, 924:1015 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 6 14 theme internalized 854:865 arg1 ZIP14 867:871 internalized ZIP14 854:871 internalized ZIP14 854:871 Iron inhibited membrane extraction of internalized ZIP14, resulting in higher steady-state levels of ZIP14. 24927598 5 15 theme protein 787:793 arg1 pathway 807:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 0 16 theme iron-regulated 3:16 arg1 pathway 70:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway 0:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14 0:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 8 17 theme membrane 1222:1229 arg1 proteins 1231:1238 endocytosed plasma membrane proteins 1203:1238 endocytosed plasma membrane proteins 1203:1238 These findings highlight the importance of proteasomes in the degradation of endocytosed plasma membrane proteins. 24927598 8 18 theme plasma 1215:1220 arg1 proteins 1231:1238 endocytosed plasma membrane proteins 1203:1238 endocytosed plasma membrane proteins 1203:1238 These findings highlight the importance of proteasomes in the degradation of endocytosed plasma membrane proteins. 24927598 5 19 theme retrograde 660:669 arg1 trafficking 671:681 the retrograde trafficking 656:681 the retrograde trafficking to the endoplasmic reticulum 656:710 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 3 20 theme multitransmembrane 402:419 arg1 domains 421:427 multitransmembrane domains 402:427 multitransmembrane domains 402:427 ZIP14 is a newly identified iron transporter with multitransmembrane domains. 24927598 6 21 theme membrane 831:838 arg1 extraction 840:849 membrane extraction 831:849 membrane extraction of internalized ZIP14 831:871 Iron inhibited membrane extraction of internalized ZIP14, resulting in higher steady-state levels of ZIP14. 24927598 7 22 link N-linked 943:950 arg1 glycosylation 953:965 Asparagine-linked (N-linked) glycosylation 924:965 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, 924:1015 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 1 23 theme Protein 127:133 arg1 degradation 135:145 Protein degradation 127:145 Protein degradation 127:145 Protein degradation is instrumental in regulating cellular function. 24927598 0 24 theme degradation 58:68 arg1 pathway 70:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway 0:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14 0:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 8 25 theme proteasomes 1169:1179 arg1 importance 1155:1164 the importance 1151:1164 the importance of proteasomes in the degradation of endocytosed plasma membrane proteins 1151:1238 These findings highlight the importance of proteasomes in the degradation of endocytosed plasma membrane proteins. 24927598 7 26 gly glycosylation 953:965 arg1 membrane 1044:1051 efficient membrane extraction 1034:1062 efficient membrane extraction of ZIP14 1034:1071 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 7 26 gly glycosylation 953:965 arg1 ZIP14 1067:1071 ZIP14 1067:1071 ZIP14 1067:1071 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 7 26 gly glycosylation 953:965 arg1 glycosylation 994:1006 particularly the glycosylation 977:1006 particularly the glycosylation at N102 977:1014 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 7 26 gly glycosylation 953:965 arg1 ZIP14 970:974 ZIP14 970:974 ZIP14 970:974 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 2 27 theme multivesicular 277:290 arg1 bodies 292:297 multivesicular bodies 277:297 multivesicular bodies 277:297 Plasma membrane proteins targeted for degradation are internalized and sorted to multivesicular bodies, which fuse with lysosomes, where they are degraded. 24927598 0 28 theme proteasomal 46:56 arg1 pathway 70:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway 0:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14 0:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 5 29 theme well-defined 741:752 arg1 pathway 807:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 the well-defined endoplasmic reticulum-associated protein degradation pathway 737:813 This pathway did not depend on the retrograde trafficking to the endoplasmic reticulum and thus did not involve the well-defined endoplasmic reticulum-associated protein degradation pathway. 24927598 4 30 theme ZIP14 504:508 arg1 levels 510:515 ZIP14 levels 504:515 ZIP14 levels 504:515 In an attempt to dissect the molecular mechanisms by which iron regulates ZIP14 levels, we found that ZIP14 is endocytosed, extracted from membranes, deglycosylated, and degraded by proteasomes. 24927598 0 31 theme glycosylation-dependent 22:44 arg1 pathway 70:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway 0:76 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14 0:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 3 32 theme iron 380:383 arg1 ZIP14 352:356 ZIP14 352:356 ZIP14 352:356 ZIP14 is a newly identified iron transporter with multitransmembrane domains. 24927598 3 32 theme iron 380:383 arg1 transporter 385:395 a newly identified iron transporter 361:395 a newly identified iron transporter with multitransmembrane domains 361:427 ZIP14 is a newly identified iron transporter with multitransmembrane domains. 24927598 7 33 theme iron 1108:1111 arg1 sensitivity 1113:1123 its iron sensitivity 1104:1123 its iron sensitivity 1104:1123 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 2 34 theme Plasma 196:201 arg1 proteins 212:219 Plasma membrane proteins 196:219 Plasma membrane proteins targeted for degradation 196:244 Plasma membrane proteins targeted for degradation are internalized and sorted to multivesicular bodies, which fuse with lysosomes, where they are degraded. 24927598 3 35 with transporter 385:395 arg1 domains 421:427 multitransmembrane domains 402:427 multitransmembrane domains 402:427 ZIP14 is a newly identified iron transporter with multitransmembrane domains. 24927598 4 36 gly deglycosylated 580:593 arg1 ZIP14 532:536 ZIP14 532:536 ZIP14 532:536 In an attempt to dissect the molecular mechanisms by which iron regulates ZIP14 levels, we found that ZIP14 is endocytosed, extracted from membranes, deglycosylated, and degraded by proteasomes. 24927598 4 37 theme molecular 459:467 arg1 mechanisms 469:478 the molecular mechanisms 455:478 the molecular mechanisms by which iron regulates ZIP14 levels 455:515 In an attempt to dissect the molecular mechanisms by which iron regulates ZIP14 levels, we found that ZIP14 is endocytosed, extracted from membranes, deglycosylated, and degraded by proteasomes. 24927598 2 38 theme membrane 203:210 arg1 proteins 212:219 Plasma membrane proteins 196:219 Plasma membrane proteins targeted for degradation 196:244 Plasma membrane proteins targeted for degradation are internalized and sorted to multivesicular bodies, which fuse with lysosomes, where they are degraded. 24927598 7 39 from N102 1011:1014 arg1 glycosylation 994:1006 particularly the glycosylation 977:1006 particularly the glycosylation at N102 977:1014 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 7 39 from N102 1011:1014 arg1 ZIP14 970:974 ZIP14 970:974 ZIP14 970:974 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 6 40 theme steady-state 894:905 arg1 levels 907:912 higher steady-state levels 887:912 higher steady-state levels of ZIP14 887:921 Iron inhibited membrane extraction of internalized ZIP14, resulting in higher steady-state levels of ZIP14. 24927598 7 41 theme ZIP14 970:974 arg1 glycosylation 953:965 Asparagine-linked (N-linked) glycosylation 924:965 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, 924:1015 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 0 42 theme plasma 86:91 arg1 transporter 108:118 the plasma membrane metal transporter ZIP14 82:124 the plasma membrane metal transporter ZIP14 82:124 An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. 24927598 6 43 theme higher 887:892 arg1 levels 907:912 higher steady-state levels 887:912 higher steady-state levels of ZIP14 887:921 Iron inhibited membrane extraction of internalized ZIP14, resulting in higher steady-state levels of ZIP14. 24927598 7 44 theme ZIP14 1067:1071 arg1 extraction 1053:1062 efficient membrane extraction 1034:1062 efficient membrane extraction of ZIP14 1034:1071 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 8 45 from importance 1155:1164 arg1 degradation 1188:1198 the degradation 1184:1198 the degradation of endocytosed plasma membrane proteins 1184:1238 These findings highlight the importance of proteasomes in the degradation of endocytosed plasma membrane proteins. 24927598 8 46 theme endocytosed 1203:1213 arg1 proteins 1231:1238 endocytosed plasma membrane proteins 1203:1238 endocytosed plasma membrane proteins 1203:1238 These findings highlight the importance of proteasomes in the degradation of endocytosed plasma membrane proteins. 24927598 7 47 theme membrane 1044:1051 arg1 extraction 1053:1062 efficient membrane extraction 1034:1062 efficient membrane extraction of ZIP14 1034:1071 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 1 48 theme cellular 177:184 arg1 function 186:193 cellular function 177:193 cellular function 177:193 Protein degradation is instrumental in regulating cellular function. 24927598 7 49 gly glycosylation 994:1006 arg1 N102 1011:1014 N102 1011:1014 N102 1011:1014 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24927598 7 49 gly glycosylation 994:1006 arg2 N102 1011:1014 N102 1011:1014 N102 1011:1014 Asparagine-linked (N-linked) glycosylation of ZIP14, particularly the glycosylation at N102, was required for efficient membrane extraction of ZIP14 and therefore is necessary for its iron sensitivity. 24256719 6 0 theme missing 980:986 arg1 link 988:991 a key missing link 974:991 a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy 974:1155 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 3 1 theme AGO61-knockout 514:527 arg1 mice 529:532 AGO61-knockout mice 514:532 AGO61-knockout mice 514:532 Here we show the phenotypes of AGO61-knockout mice and demonstrate that AGO61 is indispensable for the formation of laminin-binding glycans of α-DG. 24256719 1 2 theme laminin-binding 250:264 arg1 activity 266:273 laminin-binding activity 250:273 laminin-binding activity 250:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 6 3 theme key 976:978 arg1 link 988:991 a key missing link 974:991 a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy 974:1155 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 5 4 link threonine-linked 907:922 arg1 moieties 933:940 specific threonine-linked mannosyl moieties 898:940 specific threonine-linked mannosyl moieties of α-DG 898:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 3 5 theme mice 529:532 arg1 phenotypes 500:509 the phenotypes 496:509 the phenotypes of AGO61-knockout mice 496:532 Here we show the phenotypes of AGO61-knockout mice and demonstrate that AGO61 is indispensable for the formation of laminin-binding glycans of α-DG. 24256719 2 6 theme identified 300:309 arg1 gene 321:324 a recently identified causative gene 289:324 a recently identified causative gene for dystroglycanopathy 289:347 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 4 7 theme due 733:735 arg1 defect 726:731 a neuronal migration defect 705:731 a neuronal migration defect due to a lack of laminin-binding glycans 705:772 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 6 8 theme pathological 1111:1122 arg1 mechanisms 1124:1133 the pathological mechanisms 1107:1133 the pathological mechanisms of dystroglycanopathy 1107:1155 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 4 9 theme neuronal 707:714 arg1 defect 726:731 a neuronal migration defect 705:731 a neuronal migration defect due to a lack of laminin-binding glycans 705:772 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 5 10 theme mannosyl 924:931 arg1 moieties 933:940 specific threonine-linked mannosyl moieties 898:940 specific threonine-linked mannosyl moieties of α-DG 898:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 0 11 from formation 47:55 arg1 α-dystroglycan 82:95 α-dystroglycan 82:95 α-dystroglycan 82:95 AGO61-dependent GlcNAc modification primes the formation of functional glycans on α-dystroglycan. 24256719 6 12 theme new 1091:1093 arg1 insights 1095:1102 new insights 1091:1102 new insights 1091:1102 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 2 13 theme O-mannose 385:393 arg1 4-N-acetylglucosaminyltransferase 400:432 4-N-acetylglucosaminyltransferase 400:432 4-N-acetylglucosaminyltransferase 400:432 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 2 13 theme O-mannose 385:393 arg1 product 278:284 A product 276:284 A product of a recently identified causative gene for dystroglycanopathy 276:347 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 2 13 theme O-mannose 385:393 arg1 β-1 395:397 a protein O-mannose β-1 375:397 a protein O-mannose β-1 375:397 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 0 14 theme GlcNAc 16:21 arg1 modification 23:34 AGO61-dependent GlcNAc modification 0:34 AGO61-dependent GlcNAc modification 0:34 AGO61-dependent GlcNAc modification primes the formation of functional glycans on α-dystroglycan. 24256719 6 15 theme dystroglycanopathy 1138:1155 arg1 mechanisms 1124:1133 the pathological mechanisms 1107:1133 the pathological mechanisms of dystroglycanopathy 1107:1155 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 2 16 theme protein 377:383 arg1 4-N-acetylglucosaminyltransferase 400:432 4-N-acetylglucosaminyltransferase 400:432 4-N-acetylglucosaminyltransferase 400:432 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 2 16 theme protein 377:383 arg1 product 278:284 A product 276:284 A product of a recently identified causative gene for dystroglycanopathy 276:347 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 2 16 theme protein 377:383 arg1 β-1 395:397 a protein O-mannose β-1 375:397 a protein O-mannose β-1 375:397 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 0 17 theme AGO61-dependent 0:14 arg1 modification 23:34 AGO61-dependent GlcNAc modification 0:34 AGO61-dependent GlcNAc modification 0:34 AGO61-dependent GlcNAc modification primes the formation of functional glycans on α-dystroglycan. 24256719 2 18 theme gene 321:324 arg1 AGO61 350:354 AGO61 350:354 AGO61 350:354 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 2 18 theme gene 321:324 arg1 product 278:284 A product 276:284 A product of a recently identified causative gene for dystroglycanopathy 276:347 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 2 18 theme gene 321:324 arg1 β-1 395:397 a protein O-mannose β-1 375:397 a protein O-mannose β-1 375:397 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 5 19 theme α-DG 945:948 arg1 moieties 933:940 specific threonine-linked mannosyl moieties 898:940 specific threonine-linked mannosyl moieties of α-DG 898:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 5 20 theme AGO61-dependent 858:872 arg1 modifications 881:893 AGO61-dependent GlcNAc modifications 858:893 AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG 858:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 6 21 theme glycan 1044:1049 arg1 motif 1051:1055 the physiologically critical glycan motif 1015:1055 the physiologically critical glycan motif 1015:1055 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 2 22 theme causative 311:319 arg1 gene 321:324 a recently identified causative gene 289:324 a recently identified causative gene for dystroglycanopathy 289:347 A product of a recently identified causative gene for dystroglycanopathy, AGO61, acted in vitro as a protein O-mannose β-1, 4-N-acetylglucosaminyltransferase, although it was not functionally characterized. 24256719 1 23 theme functional 201:210 arg1 glycans 212:218 functional glycans 201:218 functional glycans on α-dystroglycan (α-DG) with laminin-binding activity 201:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 4 24 theme glycans 766:772 arg1 lack 742:745 a lack 740:745 a lack of laminin-binding glycans 740:772 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 6 25 theme critical 1035:1042 arg1 motif 1051:1055 the physiologically critical glycan motif 1015:1055 the physiologically critical glycan motif 1015:1055 These findings provide a key missing link for understanding how the physiologically critical glycan motif is displayed on α-DG and provides new insights on the pathological mechanisms of dystroglycanopathy. 24256719 1 26 with α-dystroglycan 223:236 arg1 activity 266:273 laminin-binding activity 250:273 laminin-binding activity 250:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 27 theme major 122:126 arg1 class 128:132 a major class 120:132 a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity 120:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 27 theme major 122:126 arg1 Dystroglycanopathy 98:115 Dystroglycanopathy 98:115 Dystroglycanopathy 98:115 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 28 theme glycans 212:218 arg1 deficiency 187:196 a deficiency 185:196 a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity 185:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 5 29 theme functional 814:823 arg1 glycosylation 830:842 functional α-DG glycosylation 814:842 functional α-DG glycosylation 814:842 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 5 30 theme specific 898:905 arg1 moieties 933:940 specific threonine-linked mannosyl moieties 898:940 specific threonine-linked mannosyl moieties of α-DG 898:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 4 31 theme lamina 684:689 arg1 formation 691:699 abnormal basal lamina formation 669:699 abnormal basal lamina formation 669:699 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 5 32 theme α-DG 825:828 arg1 glycosylation 830:842 functional α-DG glycosylation 814:842 functional α-DG glycosylation 814:842 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 3 33 theme α-DG 626:629 arg1 glycans 615:621 laminin-binding glycans 599:621 laminin-binding glycans of α-DG 599:629 Here we show the phenotypes of AGO61-knockout mice and demonstrate that AGO61 is indispensable for the formation of laminin-binding glycans of α-DG. 24256719 5 34 theme threonine-linked 907:922 arg1 moieties 933:940 specific threonine-linked mannosyl moieties 898:940 specific threonine-linked mannosyl moieties of α-DG 898:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 4 35 theme basal 678:682 arg1 formation 691:699 abnormal basal lamina formation 669:699 abnormal basal lamina formation 669:699 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 0 36 theme functional 60:69 arg1 glycans 71:77 functional glycans 60:77 functional glycans 60:77 AGO61-dependent GlcNAc modification primes the formation of functional glycans on α-dystroglycan. 24256719 3 37 theme laminin-binding 599:613 arg1 glycans 615:621 laminin-binding glycans 599:621 laminin-binding glycans of α-DG 599:629 Here we show the phenotypes of AGO61-knockout mice and demonstrate that AGO61 is indispensable for the formation of laminin-binding glycans of α-DG. 24256719 1 38 from α-dystroglycan 223:236 arg1 deficiency 187:196 a deficiency 185:196 a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity 185:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 3 39 theme glycans 615:621 arg1 formation 586:594 the formation 582:594 the formation of laminin-binding glycans of α-DG 582:629 Here we show the phenotypes of AGO61-knockout mice and demonstrate that AGO61 is indispensable for the formation of laminin-binding glycans of α-DG. 24256719 1 40 from glycans 212:218 arg1 α-DG 239:242 α-DG 239:242 α-DG 239:242 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 40 from glycans 212:218 arg1 α-dystroglycan 223:236 α-dystroglycan 223:236 α-dystroglycan (α-DG) with laminin-binding activity 223:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 41 theme congenital 137:146 arg1 dystrophy 157:165 congenital muscular dystrophy 137:165 congenital muscular dystrophy 137:165 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 4 42 theme AGO61-knockout 632:645 arg1 brain 653:657 AGO61-knockout mouse brain 632:657 AGO61-knockout mouse brain 632:657 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 1 43 from deficiency 187:196 arg1 α-DG 239:242 α-DG 239:242 α-DG 239:242 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 43 from deficiency 187:196 arg1 α-dystroglycan 223:236 α-dystroglycan 223:236 α-dystroglycan (α-DG) with laminin-binding activity 223:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 44 theme muscular 148:155 arg1 dystrophy 157:165 congenital muscular dystrophy 137:165 congenital muscular dystrophy 137:165 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 4 45 theme laminin-binding 750:764 arg1 glycans 766:772 laminin-binding glycans 750:772 laminin-binding glycans 750:772 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 5 46 mod modifications 881:893 arg1 moieties 933:940 specific threonine-linked mannosyl moieties 898:940 specific threonine-linked mannosyl moieties of α-DG 898:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 5 46 mod modifications 881:893 arg3 GlcNAc 874:879 AGO61-dependent GlcNAc modifications 858:893 AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG 858:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 4 47 theme abnormal 669:676 arg1 formation 691:699 abnormal basal lamina formation 669:699 abnormal basal lamina formation 669:699 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 1 48 theme dystrophy 157:165 arg1 class 128:132 a major class 120:132 a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity 120:273 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 1 48 theme dystrophy 157:165 arg1 Dystroglycanopathy 98:115 Dystroglycanopathy 98:115 Dystroglycanopathy 98:115 Dystroglycanopathy is a major class of congenital muscular dystrophy that is caused by a deficiency of functional glycans on α-dystroglycan (α-DG) with laminin-binding activity. 24256719 5 49 theme moieties 933:940 arg1 modifications 881:893 AGO61-dependent GlcNAc modifications 858:893 AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG 858:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 0 50 theme glycans 71:77 arg1 formation 47:55 the formation 43:55 the formation of functional glycans on α-dystroglycan 43:95 AGO61-dependent GlcNAc modification primes the formation of functional glycans on α-dystroglycan. 24256719 5 51 theme GlcNAc 874:879 arg1 modifications 881:893 AGO61-dependent GlcNAc modifications 858:893 AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG 858:948 Furthermore, our results indicate that functional α-DG glycosylation was primed by AGO61-dependent GlcNAc modifications of specific threonine-linked mannosyl moieties of α-DG. 24256719 4 52 theme migration 716:724 arg1 defect 726:731 a neuronal migration defect 705:731 a neuronal migration defect due to a lack of laminin-binding glycans 705:772 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 24256719 4 53 theme mouse 647:651 arg1 brain 653:657 AGO61-knockout mouse brain 632:657 AGO61-knockout mouse brain 632:657 AGO61-knockout mouse brain exhibited abnormal basal lamina formation and a neuronal migration defect due to a lack of laminin-binding glycans. 16201406 4 0 theme natural 828:834 arg1 hLF 836:838 natural hLF 828:838 natural hLF 828:838 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 1 1 theme Human 160:164 arg1 hLF 179:181 hLF 179:181 hLF 179:181 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 1 1 theme Human 160:164 arg1 lactoferrin 166:176 Human lactoferrin 160:176 Human lactoferrin (hLF) 160:182 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 1 1 theme Human 160:164 arg1 glycoprotein 203:214 an iron-binding glycoprotein 187:214 an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation 187:288 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 4 2 located found 819:823 arg1 hLF 836:838 natural hLF 828:838 natural hLF 828:838 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 2 located found 819:823 arg2 glycans 783:789 complex-type glycans 770:789 complex-type glycans 770:789 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 2 located found 819:823 arg2 glycans 811:817 the only glycans 802:817 the only glycans found on natural hLF 802:838 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 0 3 theme cows 90:93 arg1 milk 71:74 the milk 67:74 the milk of transgenic cows 67:93 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 5 4 theme Asp561 1072:1077 arg1 side-chain 1058:1067 only the side-chain 1049:1067 only the side-chain of Asp561 1049:1077 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 4 5 dep error 894:898 arg1 deviation 908:916 r.m.s. deviation 901:916 r.m.s. deviation of only 0.28 A for the main-chain atoms 901:956 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 6 theme hybrid-type 733:743 arg1 glycans 754:760 oligomannose- and hybrid-type N-linked glycans 715:760 oligomannose- and hybrid-type N-linked glycans 715:760 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 3 7 theme cows 544:547 arg1 milk 525:528 the milk 521:528 the milk of transgenic cows 521:547 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 0 8 theme transgenic 79:88 arg1 cows 90:93 transgenic cows 79:93 transgenic cows 79:93 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 3 9 theme recombinant 485:495 arg1 rhLF 502:505 rhLF 502:505 rhLF 502:505 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 3 9 theme recombinant 485:495 arg1 hLF 497:499 recombinant hLF 485:499 recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock 485:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 3 10 theme crystal 464:470 arg1 structure 472:480 the crystal structure 460:480 the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock 460:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 7 11 theme recombinant 1323:1333 arg1 proteins 1341:1348 recombinant human proteins 1323:1348 recombinant human proteins 1323:1348 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 2 12 theme production 380:389 arg1 means 371:375 alternative means 359:375 alternative means of production of this biopharmaceutical 359:415 As the availability of (human milk-derived) natural hLF is limited, alternative means of production of this biopharmaceutical are extensively researched. 16201406 3 13 theme transgenic 667:676 arg1 livestock 678:686 transgenic livestock 667:686 transgenic livestock 667:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 4 14 theme only 806:809 arg1 glycans 783:789 complex-type glycans 770:789 complex-type glycans 770:789 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 14 theme only 806:809 arg1 glycans 811:817 the only glycans 802:817 the only glycans found on natural hLF 802:838 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 15 theme N-linked 745:752 arg1 glycans 754:760 oligomannose- and hybrid-type N-linked glycans 715:760 oligomannose- and hybrid-type N-linked glycans 715:760 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 0 16 theme protein 4:10 arg1 structure 12:20 The protein structure 0:20 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows 0:93 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 3 17 theme livestock 678:686 arg1 milk 659:662 milk 659:662 milk of transgenic livestock 659:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 1 18 gly glycoprotein 203:214 arg1 lactoferrin 166:176 Human lactoferrin 160:176 Human lactoferrin (hLF) 160:182 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 1 18 gly glycoprotein 203:214 arg1 glycoprotein 203:214 an iron-binding glycoprotein 187:214 an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation 187:288 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 4 19 contain contains 706:713 arg2 glycans 754:760 oligomannose- and hybrid-type N-linked glycans 715:760 oligomannose- and hybrid-type N-linked glycans 715:760 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 19 contain contains 706:713 arg1 rhLF 701:704 rhLF 701:704 rhLF 701:704 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 1 20 theme excessive 267:275 arg1 inflammation 277:288 excessive inflammation 267:288 excessive inflammation 267:288 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 5 21 theme polymorphic 982:992 arg1 acids 1000:1004 polymorphic amino acids 982:1004 polymorphic amino acids between the natural and rhLF variant used 982:1046 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 5 22 from differences 967:977 arg1 acids 1000:1004 polymorphic amino acids 982:1004 polymorphic amino acids between the natural and rhLF variant used 982:1046 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 2 23 theme alternative 359:369 arg1 means 371:375 alternative means 359:375 alternative means of production of this biopharmaceutical 359:415 As the availability of (human milk-derived) natural hLF is limited, alternative means of production of this biopharmaceutical are extensively researched. 16201406 5 24 theme rhLF 1105:1108 arg1 map 1127:1129 the rhLF electron density map 1101:1129 the rhLF electron density map 1101:1129 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 4 25 theme experimental 881:892 arg1 error 894:898 the experimental error 877:898 the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms) 877:957 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 7 26 theme mammary 1285:1291 arg1 gland 1293:1297 the transgenic cow mammary gland 1266:1297 the transgenic cow mammary gland 1266:1297 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 2 27 dep natural 335:341 arg1 milk-derived 321:332 milk-derived 321:332 milk-derived 321:332 As the availability of (human milk-derived) natural hLF is limited, alternative means of production of this biopharmaceutical are extensively researched. 16201406 5 28 theme electron 1110:1117 arg1 map 1127:1129 the rhLF electron density map 1101:1129 the rhLF electron density map 1101:1129 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 3 29 theme reported 603:610 arg1 resolution 554:563 a resolution 552:563 a resolution of 2.4 A. To our knowledge 552:590 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 3 29 theme reported 603:610 arg1 structure 612:620 the first reported structure 593:620 the first reported structure of a recombinant protein produced in milk of transgenic livestock 593:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 2 30 theme hLF 343:345 arg1 availability 298:309 the availability 294:309 the availability of (human milk-derived) natural hLF 294:345 As the availability of (human milk-derived) natural hLF is limited, alternative means of production of this biopharmaceutical are extensively researched. 16201406 2 31 theme biopharmaceutical 399:415 arg1 production 380:389 production 380:389 production of this biopharmaceutical 380:415 As the availability of (human milk-derived) natural hLF is limited, alternative means of production of this biopharmaceutical are extensively researched. 16201406 0 32 theme human 37:41 arg1 lactoferrin 43:53 recombinant human lactoferrin 25:53 recombinant human lactoferrin produced in the milk of transgenic cows 25:93 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 4 33 link N-linked 745:752 arg1 glycans 754:760 oligomannose- and hybrid-type N-linked glycans 715:760 oligomannose- and hybrid-type N-linked glycans 715:760 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 0 34 theme human 128:132 arg1 lactoferrin 147:157 human milk-derived lactoferrin 128:157 human milk-derived lactoferrin 128:157 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 5 35 theme natural 1018:1024 arg1 variant 1035:1041 the natural and rhLF variant 1014:1041 the natural and rhLF variant used 1014:1046 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 7 36 theme gland 1293:1297 arg1 validity 1254:1261 the validity 1250:1261 the validity of the transgenic cow mammary gland 1250:1297 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 7 36 theme gland 1293:1297 arg1 vehicle 1304:1310 a vehicle 1302:1310 a vehicle to produce recombinant human proteins 1302:1348 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 0 37 link milk-derived 134:145 arg1 lactoferrin 147:157 human milk-derived lactoferrin 128:157 human milk-derived lactoferrin 128:157 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 1 38 theme iron-binding 190:201 arg1 lactoferrin 166:176 Human lactoferrin 160:176 Human lactoferrin (hLF) 160:182 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 1 38 theme iron-binding 190:201 arg1 glycoprotein 203:214 an iron-binding glycoprotein 187:214 an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation 187:288 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 16201406 0 39 theme recombinant 25:35 arg1 lactoferrin 43:53 recombinant human lactoferrin 25:53 recombinant human lactoferrin produced in the milk of transgenic cows 25:93 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 4 40 theme main-chain 941:950 arg1 atoms 952:956 the main-chain atoms 937:956 the main-chain atoms 937:956 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 3 41 theme A. 572:573 arg1 resolution 554:563 a resolution 552:563 a resolution of 2.4 A. To our knowledge 552:590 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 3 41 theme A. 572:573 arg1 structure 612:620 the first reported structure 593:620 the first reported structure of a recombinant protein produced in milk of transgenic livestock 593:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 5 42 theme density 1119:1125 arg1 map 1127:1129 the rhLF electron density map 1101:1129 the rhLF electron density map 1101:1129 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 4 43 theme oligomannose- 715:727 arg1 glycans 754:760 oligomannose- and hybrid-type N-linked glycans 715:760 oligomannose- and hybrid-type N-linked glycans 715:760 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 0 44 theme lactoferrin 147:157 arg1 structure 115:123 the structure 111:123 the structure of human milk-derived lactoferrin 111:157 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 5 45 theme rhLF 1030:1033 arg1 variant 1035:1041 the natural and rhLF variant 1014:1041 the natural and rhLF variant used 1014:1046 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 7 46 theme cow 1281:1283 arg1 gland 1293:1297 the transgenic cow mammary gland 1266:1297 the transgenic cow mammary gland 1266:1297 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 2 47 theme natural 335:341 arg1 hLF 343:345 (human milk-derived) natural hLF 314:345 (human milk-derived) natural hLF 314:345 As the availability of (human milk-derived) natural hLF is limited, alternative means of production of this biopharmaceutical are extensively researched. 16201406 0 48 theme lactoferrin 43:53 arg1 structure 12:20 The protein structure 0:20 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows 0:93 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 0 49 theme milk-derived 134:145 arg1 lactoferrin 147:157 human milk-derived lactoferrin 128:157 human milk-derived lactoferrin 128:157 The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin. 16201406 3 50 theme recombinant 627:637 arg1 protein 639:645 a recombinant protein 625:645 a recombinant protein produced in milk of transgenic livestock 625:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 5 51 theme amino 994:998 arg1 acids 1000:1004 polymorphic amino acids 982:1004 polymorphic amino acids between the natural and rhLF variant used 982:1046 Of the differences in polymorphic amino acids between the natural and rhLF variant used, only the side-chain of Asp561 could be modeled into the rhLF electron density map. 16201406 3 52 theme hLF 497:499 arg1 structure 472:480 the crystal structure 460:480 the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock 460:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 6 53 theme variant 1205:1211 arg1 integrity 1183:1191 the structural integrity 1168:1191 the structural integrity of the rhLF variant used in this study 1168:1230 Taken together, the results confirm the structural integrity of the rhLF variant used in this study. 16201406 3 54 theme protein 639:645 arg1 resolution 554:563 a resolution 552:563 a resolution of 2.4 A. To our knowledge 552:590 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 3 54 theme protein 639:645 arg1 structure 612:620 the first reported structure 593:620 the first reported structure of a recombinant protein produced in milk of transgenic livestock 593:686 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 6 55 theme structural 1172:1181 arg1 integrity 1183:1191 the structural integrity 1168:1191 the structural integrity of the rhLF variant used in this study 1168:1230 Taken together, the results confirm the structural integrity of the rhLF variant used in this study. 16201406 3 56 theme transgenic 533:542 arg1 cows 544:547 transgenic cows 533:547 transgenic cows 533:547 Here we report the crystal structure of recombinant hLF (rhLF) expressed in the milk of transgenic cows at a resolution of 2.4 A. To our knowledge, the first reported structure of a recombinant protein produced in milk of transgenic livestock. 16201406 7 57 theme human 1335:1339 arg1 proteins 1341:1348 recombinant human proteins 1323:1348 recombinant human proteins 1323:1348 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 7 58 theme transgenic 1270:1279 arg1 gland 1293:1297 the transgenic cow mammary gland 1266:1297 the transgenic cow mammary gland 1266:1297 It also confirms the validity of the transgenic cow mammary gland as a vehicle to produce recombinant human proteins. 16201406 4 59 theme r.m.s. 901:906 arg1 deviation 908:916 r.m.s. deviation 901:916 r.m.s. deviation of only 0.28 A for the main-chain atoms 901:956 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 60 theme A 931:931 arg1 deviation 908:916 r.m.s. deviation 901:916 r.m.s. deviation of only 0.28 A for the main-chain atoms 901:956 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 61 theme complex-type 770:781 arg1 glycans 783:789 complex-type glycans 770:789 complex-type glycans 770:789 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 4 61 theme complex-type 770:781 arg1 glycans 811:817 the only glycans 802:817 the only glycans found on natural hLF 802:838 Even though rhLF contains oligomannose- and hybrid-type N-linked glycans next to complex-type glycans, which are the only glycans found on natural hLF, the structures are identical within the experimental error (r.m.s. deviation of only 0.28 A for the main-chain atoms). 16201406 6 62 theme rhLF 1200:1203 arg1 variant 1205:1211 the rhLF variant 1196:1211 the rhLF variant used in this study 1196:1230 Taken together, the results confirm the structural integrity of the rhLF variant used in this study. 16201406 1 63 theme host 232:235 arg1 defence 237:243 the host defence 228:243 the host defence against infection and excessive inflammation 228:288 Human lactoferrin (hLF) is an iron-binding glycoprotein involved in the host defence against infection and excessive inflammation. 1581307 9 0 theme protein 1829:1835 arg1 structure 1837:1845 the protein structure 1825:1845 the protein structure 1825:1845 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 5 1 from sites 1060:1064 arg1 different 1031:1039 different 1031:1039 different 1031:1039 The copper coordination is different in the two binding sites. 1581307 5 1 from sites 1060:1064 arg1 coordination 1015:1026 The copper coordination 1004:1026 The copper coordination 1004:1026 The copper coordination is different in the two binding sites. 1581307 9 2 theme small 1736:1740 arg1 movements 1742:1750 small movements 1736:1750 small movements of the metal ion and anion within each binding site, which do not affect the protein structure 1736:1845 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 8 3 theme diferric 1435:1442 arg1 Fe2Lf 1453:1457 Fe2Lf 1453:1457 Fe2Lf 1453:1457 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 8 3 theme diferric 1435:1442 arg1 protein 1444:1450 the diferric protein 1431:1450 the diferric protein 1431:1450 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 1 4 theme copper-lactoferrin 244:261 arg1 analysis 226:233 crystallographic analysis 209:233 crystallographic analysis of human copper-lactoferrin, Cu2Lf 209:268 The structural consequences of binding a metal other than iron to a transferrin have been examined by crystallographic analysis of human copper-lactoferrin, Cu2Lf. 1581307 0 5 theme copper-lactoferrin 67:84 arg1 structure 48:56 the crystal structure 36:56 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 0:105 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 0 6 from substitution 6:17 arg1 transferrins 22:33 transferrins 22:33 transferrins 22:33 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 3 7 theme restrained 503:512 arg1 methods 528:534 restrained least-squares methods 503:534 restrained least-squares methods 503:534 The structure was refined crystallographically, by restrained least-squares methods, starting with a model based on the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted. 1581307 9 8 dep ion 1765:1767 arg1 the 1755:1757 the 1755:1757 the 1755:1757 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 6 9 theme apical 1213:1218 arg1 bond 1220:1223 a longer apical bond 1204:1223 a longer apical bond to Tyr 92 1204:1233 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 4 10 theme lengths 859:865 arg1 deviation 841:849 rms deviation 837:849 rms deviation of bond lengths from standard values of 0.018 A 837:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 11 theme bi 766:767 arg1 carbonate 769:777 2 (bi)carbonate 763:777 2 (bi)carbonate ions 763:782 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 0 12 theme 2.1-A 89:93 arg1 resolution 95:104 2.1-A resolution 89:104 2.1-A resolution 89:104 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 4 13 theme solvent 793:799 arg1 molecules 801:809 308 solvent molecules 789:809 308 solvent molecules 789:809 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 6 14 theme N-terminal 1074:1083 arg1 site 1085:1088 the N-terminal site 1070:1088 the N-terminal site 1070:1088 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 4 15 theme final 691:695 arg1 model 697:701 The final model 687:701 The final model 687:701 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 16 theme carbonate 769:777 arg1 ions 779:782 2 (bi)carbonate ions 763:782 2 (bi)carbonate ions 763:782 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 9 17 theme metal 1759:1763 arg1 ion 1765:1767 metal ion 1759:1767 metal ion 1759:1767 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 2 18 theme diffraction 277:287 arg1 data 289:292 X-ray diffraction data 271:292 X-ray diffraction data 271:292 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 8 19 theme protein 1498:1504 arg1 domains 1506:1512 the protein domains 1494:1512 the protein domains over the metal 1494:1527 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 2 20 theme oscillation 382:392 arg1 photography 394:404 oscillation photography 382:404 oscillation photography 382:404 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 4 21 theme standard 872:879 arg1 values 881:886 standard values 872:886 standard values of 0.018 A 872:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 5 22 theme copper 1008:1013 arg1 different 1031:1039 different 1031:1039 different 1031:1039 The copper coordination is different in the two binding sites. 1581307 5 22 theme copper 1008:1013 arg1 coordination 1015:1026 The copper coordination 1004:1026 The copper coordination 1004:1026 The copper coordination is different in the two binding sites. 1581307 4 23 from values 881:886 arg1 deviation 841:849 rms deviation 837:849 rms deviation of bond lengths from standard values of 0.018 A 837:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 9 24 theme ion 1765:1767 arg1 movements 1742:1750 small movements 1736:1750 small movements of the metal ion and anion within each binding site, which do not affect the protein structure 1736:1845 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 2 25 theme X-ray 271:275 arg1 data 289:292 X-ray diffraction data 271:292 X-ray diffraction data 271:292 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 4 26 contain has 811:813 arg1 model 697:701 The final model 687:701 The final model 687:701 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 26 contain has 811:813 arg2 stereochemistry 820:834 good stereochemistry 815:834 good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) 815:898 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 2 27 theme 2.1-A 434:438 arg1 resolution 440:449 2.1-A resolution 434:449 2.1-A resolution 434:449 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 4 28 theme A 897:897 arg1 values 881:886 standard values 872:886 standard values of 0.018 A 872:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 29 theme 0.018 891:895 arg1 A 897:897 0.018 A 891:897 0.018 A 891:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 8 30 theme protein 1394:1400 arg1 structure 1402:1410 The protein structure 1390:1410 The protein structure 1390:1410 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 8 30 theme protein 1394:1400 arg1 same 1419:1422 same 1419:1422 same 1419:1422 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 0 31 theme Metal 0:4 arg1 substitution 6:17 Metal substitution 0:17 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 0:105 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 6 32 theme equatorial 1130:1139 arg1 bonds 1141:1145 equatorial bonds 1130:1145 equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion 1130:1198 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 9 33 theme anion 1773:1777 arg1 movements 1742:1750 small movements 1736:1750 small movements of the metal ion and anion within each binding site, which do not affect the protein structure 1736:1845 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 4 34 theme R 929:929 arg1 value 931:935 a crystallographic R value 910:935 a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution 910:1001 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 6 35 theme monodentate 1182:1192 arg1 anion 1194:1198 a monodentate anion 1180:1198 a monodentate anion 1180:1198 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 4 36 theme 0.196 940:944 arg1 value 931:935 a crystallographic R value 910:935 a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution 910:1001 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 6 37 theme square 1107:1112 arg1 geometry 1095:1102 the geometry 1091:1102 the geometry 1091:1102 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 6 37 theme square 1107:1112 arg1 pyramidal 1114:1122 square pyramidal 1107:1122 square pyramidal 1107:1122 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 7 38 theme bidentate 1373:1381 arg1 anion 1383:1387 an asymmetrically bidentate anion 1355:1387 an asymmetrically bidentate anion 1355:1387 In the C-terminal site, the geometry is distorted octahedral, with bonds to Asp 395, Tyr 435, Tyr 528, and His 597 and an asymmetrically bidentate anion. 1581307 2 39 theme synchrotron 411:421 arg1 source 423:428 a synchrotron source 409:428 a synchrotron source 409:428 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 7 40 theme C-terminal 1243:1252 arg1 site 1254:1257 the C-terminal site 1239:1257 the C-terminal site 1239:1257 In the C-terminal site, the geometry is distorted octahedral, with bonds to Asp 395, Tyr 435, Tyr 528, and His 597 and an asymmetrically bidentate anion. 1581307 4 41 theme rms 837:839 arg1 deviation 841:849 rms deviation 837:849 rms deviation of bond lengths from standard values of 0.018 A 837:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 42 theme protein 720:726 arg1 atoms 728:732 5321 protein atoms 715:732 5321 protein atoms (691 residues) 715:747 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 42 theme protein 720:726 arg1 residues 739:746 691 residues 735:746 691 residues 735:746 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 2 43 theme 2.6-A 360:364 arg1 resolution 366:375 2.6-A resolution 360:375 2.6-A resolution 360:375 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 4 44 theme bond 854:857 arg1 lengths 859:865 bond lengths 854:865 bond lengths 854:865 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 9 45 theme binding 1791:1797 arg1 site 1799:1802 each binding site 1786:1802 each binding site 1786:1802 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 8 46 theme domains 1506:1512 arg1 closure 1483:1489 the closure 1479:1489 the closure of the protein domains over the metal 1479:1527 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 8 46 theme domains 1506:1512 arg1 same 1536:1539 same 1536:1539 same 1536:1539 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 9 47 from differences 1686:1696 arg1 coordination 1707:1718 metal coordination 1701:1718 metal coordination 1701:1718 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 6 48 from pyramidal 1114:1122 arg1 site 1085:1088 the N-terminal site 1070:1088 the N-terminal site 1070:1088 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 3 49 theme metal 627:631 arg1 ions 633:636 metal ions 627:636 metal ions 627:636 The structure was refined crystallographically, by restrained least-squares methods, starting with a model based on the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted. 1581307 8 50 from same 1536:1539 arg1 case 1549:1552 each case 1544:1552 each case irrespective of whether Fe3+ or Cu2+ is bound 1544:1598 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 4 51 theme crystallographic 912:927 arg1 value 931:935 a crystallographic R value 910:935 a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution 910:1001 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 5 52 theme binding 1052:1058 arg1 sites 1060:1064 the two binding sites 1044:1064 the two binding sites 1044:1064 The copper coordination is different in the two binding sites. 1581307 4 53 theme good 815:818 arg1 stereochemistry 820:834 good stereochemistry 815:834 good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) 815:898 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 3 54 theme isomorphous 572:582 arg1 structure 593:601 the isomorphous diferric structure 568:601 the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted 568:684 The structure was refined crystallographically, by restrained least-squares methods, starting with a model based on the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted. 1581307 5 55 from different 1031:1039 arg1 sites 1060:1064 the two binding sites 1044:1064 the two binding sites 1044:1064 The copper coordination is different in the two binding sites. 1581307 1 56 theme structural 111:120 arg1 consequences 122:133 The structural consequences 107:133 The structural consequences of binding a metal other than iron to a transferrin 107:185 The structural consequences of binding a metal other than iron to a transferrin have been examined by crystallographic analysis of human copper-lactoferrin, Cu2Lf. 1581307 0 57 theme crystal 40:46 arg1 structure 48:56 the crystal structure 36:56 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 0:105 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 3 58 theme solvent 651:657 arg1 molecules 659:667 solvent molecules 651:667 solvent molecules 651:667 The structure was refined crystallographically, by restrained least-squares methods, starting with a model based on the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted. 1581307 3 59 theme diferric 584:591 arg1 structure 593:601 the isomorphous diferric structure 568:601 the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted 568:684 The structure was refined crystallographically, by restrained least-squares methods, starting with a model based on the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted. 1581307 9 60 theme metal 1701:1705 arg1 coordination 1707:1718 metal coordination 1701:1718 metal coordination 1701:1718 The differences in metal coordination are achieved by small movements of the metal ion and anion within each binding site, which do not affect the protein structure. 1581307 4 61 theme range 976:980 arg1 resolution 992:1001 the range 7.5-2.1-A resolution 972:1001 the range 7.5-2.1-A resolution 972:1001 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 8 62 theme irrespective 1554:1565 arg1 case 1549:1552 each case 1544:1552 each case irrespective of whether Fe3+ or Cu2+ is bound 1544:1598 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 1 63 theme crystallographic 209:224 arg1 analysis 226:233 crystallographic analysis 209:233 crystallographic analysis of human copper-lactoferrin, Cu2Lf 209:268 The structural consequences of binding a metal other than iron to a transferrin have been examined by crystallographic analysis of human copper-lactoferrin, Cu2Lf. 1581307 0 64 from resolution 95:104 arg1 structure 48:56 the crystal structure 36:56 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 0:105 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 6 65 with pyramidal 1114:1122 arg1 bonds 1141:1145 equatorial bonds 1130:1145 equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion 1130:1198 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 6 65 with pyramidal 1114:1122 arg1 bond 1220:1223 a longer apical bond 1204:1223 a longer apical bond to Tyr 92 1204:1233 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 2 66 theme Cu2Lf 326:330 arg1 crystals 314:321 crystals 314:321 crystals of Cu2Lf 314:330 X-ray diffraction data were collected from crystals of Cu2Lf, using a diffractometer, to 2.6-A resolution, and oscillation photography on a synchrotron source, to 2.1-A resolution. 1581307 6 67 theme longer 1206:1211 arg1 bond 1220:1223 a longer apical bond 1204:1223 a longer apical bond to Tyr 92 1204:1233 In the N-terminal site, the geometry is square pyramidal, with equatorial bonds to Asp 60, Tyr 192, His 253, and a monodentate anion and a longer apical bond to Tyr 92. 1581307 0 68 theme human 61:65 arg1 copper-lactoferrin 67:84 human copper-lactoferrin 61:84 human copper-lactoferrin 61:84 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 0 69 dep substitution 6:17 arg1 structure 48:56 the crystal structure 36:56 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 0:105 Metal substitution in transferrins: the crystal structure of human copper-lactoferrin at 2.1-A resolution. 1581307 8 70 from case 1549:1552 arg1 closure 1483:1489 the closure 1479:1489 the closure of the protein domains over the metal 1479:1527 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 8 70 from case 1549:1552 arg1 same 1536:1539 same 1536:1539 same 1536:1539 The protein structure is the same as for the diferric protein, Fe2Lf, demonstrating that the closure of the protein domains over the metal is the same in each case irrespective of whether Fe3+ or Cu2+ is bound and that copper could be transported and delivered to cells equally well as iron. 1581307 3 71 theme least-squares 514:526 arg1 methods 528:534 restrained least-squares methods 503:534 restrained least-squares methods 503:534 The structure was refined crystallographically, by restrained least-squares methods, starting with a model based on the isomorphous diferric structure from which the ligands, metal ions, anions, and solvent molecules had been deleted. 1581307 4 72 theme 7.5-2.1-A 982:990 arg1 resolution 992:1001 the range 7.5-2.1-A resolution 972:1001 the range 7.5-2.1-A resolution 972:1001 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 73 dep stereochemistry 820:834 arg1 deviation 841:849 rms deviation 837:849 rms deviation of bond lengths from standard values of 0.018 A 837:897 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 74 theme Cu2+ 752:755 arg1 ions 757:760 2 Cu2+ ions 750:760 2 Cu2+ ions 750:760 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 4 75 from reflections 957:967 arg1 resolution 992:1001 the range 7.5-2.1-A resolution 972:1001 the range 7.5-2.1-A resolution 972:1001 The final model, comprising 5321 protein atoms (691 residues), 2 Cu2+ ions, 2 (bi)carbonate ions, and 308 solvent molecules has good stereochemistry (rms deviation of bond lengths from standard values of 0.018 A) and gives a crystallographic R value of 0.196 for 43,525 reflections in the range 7.5-2.1-A resolution. 1581307 1 76 theme human 238:242 arg1 Cu2Lf 264:268 Cu2Lf 264:268 Cu2Lf 264:268 The structural consequences of binding a metal other than iron to a transferrin have been examined by crystallographic analysis of human copper-lactoferrin, Cu2Lf. 1581307 1 76 theme human 238:242 arg1 copper-lactoferrin 244:261 human copper-lactoferrin 238:261 human copper-lactoferrin 238:261 The structural consequences of binding a metal other than iron to a transferrin have been examined by crystallographic analysis of human copper-lactoferrin, Cu2Lf. 15299793 4 0 theme structural 737:746 arg1 similarity 748:757 A close structural similarity 729:757 A close structural similarity 729:757 A close structural similarity is seen between the two lobes of the molecule, with differences mainly in loops and turns. 15299793 4 1 from differences 811:821 arg1 loops 833:837 loops 833:837 loops 833:837 A close structural similarity is seen between the two lobes of the molecule, with differences mainly in loops and turns. 15299793 4 1 from differences 811:821 arg1 turns 843:847 turns 843:847 turns 843:847 A close structural similarity is seen between the two lobes of the molecule, with differences mainly in loops and turns. 15299793 8 2 from atom 1669:1672 arg1 lobe 1682:1685 each lobe 1677:1685 each lobe 1677:1685 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 8 2 from atom 1669:1672 arg1 present 1643:1649 present 1643:1649 present 1643:1649 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 9 3 theme glycan 1828:1833 arg1 six 1810:1812 six 1810:1812 six 1810:1812 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 9 3 theme glycan 1828:1833 arg1 glycan 1828:1833 the C-lobe glycan 1817:1833 the C-lobe glycan 1817:1833 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 8 4 theme water 1625:1629 arg1 molecule 1631:1638 one tightly bound water molecule 1607:1638 one tightly bound water molecule 1607:1638 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 5 5 from binding 968:974 arg1 site 1084:1087 the N-lobe site 1073:1087 the N-lobe site relative to the C-lobe site 1073:1115 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 3 6 theme final 626:630 arg1 0.179 661:665 0.179 661:665 0.179 661:665 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 3 6 theme final 626:630 arg1 R-factor 649:656 the final crystallographic R-factor 622:656 the final crystallographic R-factor 622:656 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 2 7 theme restrained 280:289 arg1 refinement 305:314 Following restrained least-squares refinement 270:314 Following restrained least-squares refinement 270:314 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 6 8 theme sites 1293:1297 arg1 stability 1266:1274 the stability 1262:1274 the stability of the two metal sites 1262:1297 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 7 9 theme known 1415:1419 arg1 flexibility 1421:1431 the known flexibility 1411:1431 the known flexibility of the molecule 1411:1447 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 2 10 theme carbohydrate 456:467 arg1 residues 488:495 eight sugar residues 476:495 eight sugar residues 476:495 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 2 10 theme carbohydrate 456:467 arg1 atoms 469:473 98 carbohydrate atoms 453:473 98 carbohydrate atoms (eight sugar residues) 453:496 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 4 11 theme molecule 796:803 arg1 lobes 783:787 the two lobes 775:787 the two lobes of the molecule 775:803 A close structural similarity is seen between the two lobes of the molecule, with differences mainly in loops and turns. 15299793 4 11 theme molecule 796:803 arg1 molecule 796:803 the molecule 792:803 the molecule 792:803 A close structural similarity is seen between the two lobes of the molecule, with differences mainly in loops and turns. 15299793 5 12 from bond 1038:1041 arg1 site 1084:1087 the N-lobe site 1073:1087 the N-lobe site relative to the C-lobe site 1073:1115 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 3 13 theme Root-mean-square 499:514 arg1 A 560:560 0.015 A 554:560 0.015 A for bond lengths 554:577 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 3 13 theme Root-mean-square 499:514 arg1 deviations 516:525 Root-mean-square deviations 499:525 Root-mean-square deviations from standard geometry 499:548 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 1 14 theme data 199:202 arg1 set 265:267 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 2 15 theme sugar 482:486 arg1 residues 488:495 eight sugar residues 476:495 eight sugar residues 476:495 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 2 15 theme sugar 482:486 arg1 atoms 469:473 98 carbohydrate atoms 453:473 98 carbohydrate atoms (eight sugar residues) 453:496 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 8 16 theme Fe 1666:1667 arg1 atom 1669:1672 the Fe atom 1662:1672 the Fe atom in each lobe 1662:1685 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 1 17 theme form 117:120 arg1 structure 91:99 The three-dimensional structure 69:99 The three-dimensional structure of the diferric form of human lactoferrin 69:141 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 6 18 theme metal 1287:1291 arg1 sites 1293:1297 the two metal sites 1279:1297 the two metal sites 1279:1297 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 10 19 theme direct 1845:1850 arg1 contact 1873:1879 Only one direct protein-carbohydrate contact 1836:1879 Only one direct protein-carbohydrate contact 1836:1879 Only one direct protein-carbohydrate contact can be found. 15299793 5 20 theme relative 1089:1096 arg1 site 1084:1087 the N-lobe site 1073:1087 the N-lobe site relative to the C-lobe site 1073:1115 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 1 21 theme human 125:129 arg1 lactoferrin 131:141 human lactoferrin 125:141 human lactoferrin 125:141 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 8 22 dep present 1643:1649 arg1 A 1655:1655 3.8 A 1651:1655 3.8 A 1651:1655 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 0 23 theme A 55:55 arg1 resolution 57:66 2.2 A resolution 51:66 2.2 A resolution 51:66 Structure of human diferric lactoferrin refined at 2.2 A resolution. 15299793 8 24 theme large 1556:1560 arg1 clusters 1562:1569 two large clusters 1552:1569 two large clusters (in the two interdomain clefts) 1552:1601 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 3 25 theme resolution 701:710 arg1 range 712:716 the resolution range 8.0-2.2 A 697:726 the resolution range 8.0-2.2 A 697:726 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 5 26 theme carbonate 983:991 arg1 ion 993:995 the carbonate ion 979:995 the carbonate ion 979:995 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 7 27 from number 1379:1384 arg1 few 1372:1374 few 1372:1374 few 1372:1374 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 2 28 theme solvent 431:437 arg1 molecules 439:447 469 solvent molecules 427:447 469 solvent molecules 427:447 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 5 29 theme N-lobe 1077:1082 arg1 site 1084:1087 the N-lobe site 1073:1087 the N-lobe site relative to the C-lobe site 1073:1115 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 30 theme more 942:945 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 30 theme more 942:945 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 30 theme more 942:945 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 30 theme more 942:945 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 30 theme more 942:945 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 7 31 theme interlobe 1328:1336 arg1 interactions 1338:1349 interdomain and interlobe interactions 1312:1349 interactions 1338:1349 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 5 32 theme bidentate 958:966 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 32 theme bidentate 958:966 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 32 theme bidentate 958:966 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 32 theme bidentate 958:966 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 32 theme bidentate 958:966 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 8 33 theme interdomain 1583:1593 arg1 clefts 1595:1600 the two interdomain clefts 1575:1600 the two interdomain clefts 1575:1600 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 9 34 theme limited 1750:1756 arg1 extent 1758:1763 a limited extent 1748:1763 a limited extent 1748:1763 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 3 35 from geometry 541:548 arg1 A 560:560 0.015 A 554:560 0.015 A for bond lengths 554:577 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 3 35 from geometry 541:548 arg1 deviations 516:525 Root-mean-square deviations 499:525 Root-mean-square deviations from standard geometry 499:548 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 2 36 theme protein 368:374 arg1 atoms 376:380 5330 protein atoms 363:380 5330 protein atoms (691 residues) 363:395 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 2 36 theme protein 368:374 arg1 residues 387:394 691 residues 383:394 691 residues 383:394 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 6 37 theme iron 1227:1230 arg1 site 1232:1235 the iron site 1223:1235 the iron site 1223:1235 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 3 38 dep range 712:716 arg1 A 726:726 A 726:726 the resolution range 8.0-2.2 A 697:726 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 0 39 theme human 13:17 arg1 lactoferrin 28:38 human diferric lactoferrin 13:38 human diferric lactoferrin 13:38 Structure of human diferric lactoferrin refined at 2.2 A resolution. 15299793 1 40 theme data 260:263 arg1 set 265:267 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 9 41 theme sugar 1770:1774 arg1 residues 1776:1783 two sugar residues 1766:1783 two sugar residues of the N-lobe glycan 1766:1804 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 9 41 theme sugar 1770:1774 arg1 glycan 1799:1804 the N-lobe glycan 1788:1804 the N-lobe glycan 1788:1804 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 0 42 theme lactoferrin 28:38 arg1 Structure 0:8 Structure 0:8 Structure of human diferric lactoferrin 0:38 Structure of human diferric lactoferrin refined at 2.2 A resolution. 15299793 2 43 theme 2CO 410:412 arg1 ions 421:424 2Fe(3+) and 2CO(3)(2-) ions 398:424 2Fe(3+) and 2CO(3)(2-) ions 398:424 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 9 44 theme N-lobe 1792:1797 arg1 glycan 1799:1804 the N-lobe glycan 1788:1804 the N-lobe glycan 1788:1804 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 7 45 from few 1372:1374 arg1 number 1379:1384 number 1379:1384 number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements 1379:1489 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 5 46 theme Tyr 1057:1059 arg1 ligands 1061:1067 the Tyr ligands 1053:1067 the Tyr ligands 1053:1067 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 47 theme binding 858:864 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 47 theme binding 858:864 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 47 theme binding 858:864 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 47 theme binding 858:864 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 47 theme binding 858:864 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 2 48 theme final 341:345 arg1 model 347:351 the final model 337:351 the final model 337:351 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 5 49 theme Fe-O 1033:1036 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 49 theme Fe-O 1033:1036 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 49 theme Fe-O 1033:1036 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 49 theme Fe-O 1033:1036 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 4 50 theme close 731:735 arg1 similarity 748:757 A close structural similarity 729:757 A close structural similarity 729:757 A close structural similarity is seen between the two lobes of the molecule, with differences mainly in loops and turns. 15299793 9 51 theme C-lobe 1821:1826 arg1 glycan 1828:1833 the C-lobe glycan 1817:1833 the C-lobe glycan 1817:1833 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 8 52 theme bound 1619:1623 arg1 molecule 1631:1638 one tightly bound water molecule 1607:1638 one tightly bound water molecule 1607:1638 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 5 53 dep similar 886:892 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 53 dep similar 886:892 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 53 dep similar 886:892 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 53 dep similar 886:892 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 53 dep similar 886:892 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 2 54 theme least-squares 291:303 arg1 refinement 305:314 Following restrained least-squares refinement 270:314 Following restrained least-squares refinement 270:314 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 1 55 theme A 167:167 arg1 resolution 169:178 2.2 A resolution 163:178 2.2 A resolution 163:178 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 8 56 located found 1521:1525 arg1 sites 1539:1543 discrete sites 1530:1543 discrete sites 1530:1543 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 8 56 located found 1521:1525 arg2 molecules 1507:1515 Internal water molecules 1492:1515 Internal water molecules 1492:1515 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 3 57 theme crystallographic 632:647 arg1 0.179 661:665 0.179 661:665 0.179 661:665 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 3 57 theme crystallographic 632:647 arg1 R-factor 649:656 the final crystallographic R-factor 622:656 the final crystallographic R-factor 622:656 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 2 58 theme Following 270:278 arg1 refinement 305:314 Following restrained least-squares refinement 270:314 Following restrained least-squares refinement 270:314 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 8 59 theme Internal 1492:1499 arg1 molecules 1507:1515 Internal water molecules 1492:1515 Internal water molecules 1492:1515 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 8 60 theme water 1501:1505 arg1 molecules 1507:1515 Internal water molecules 1492:1515 Internal water molecules 1492:1515 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 1 61 theme synchrotron 187:197 arg1 data 199:202 synchrotron data 187:202 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 6 62 theme Distinct 1118:1125 arg1 differences 1127:1137 Distinct differences 1118:1137 Distinct differences 1118:1137 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 1 63 theme diferric 108:115 arg1 form 117:120 the diferric form 104:120 the diferric form of human lactoferrin 104:141 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 10 64 theme protein-carbohydrate 1852:1871 arg1 contact 1873:1879 Only one direct protein-carbohydrate contact 1836:1879 Only one direct protein-carbohydrate contact 1836:1879 Only one direct protein-carbohydrate contact can be found. 15299793 3 65 theme standard 532:539 arg1 geometry 541:548 standard geometry 532:548 standard geometry 532:548 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 8 66 theme discrete 1530:1537 arg1 sites 1539:1543 discrete sites 1530:1543 discrete sites 1530:1543 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 8 67 from clusters 1562:1569 arg1 clefts 1595:1600 the two interdomain clefts 1575:1600 the two interdomain clefts 1575:1600 Internal water molecules are found in discrete sites and in two large clusters (in the two interdomain clefts) and one tightly bound water molecule is present 3.8 A from the Fe atom in each lobe. 15299793 5 68 theme apparent 904:911 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 68 theme apparent 904:911 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 68 theme apparent 904:911 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 1 69 theme lactoferrin 131:141 arg1 form 117:120 the diferric form 104:120 the diferric form of human lactoferrin 104:141 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 6 70 theme cationic 1180:1187 arg1 Lys546 1208:1213 Lys546 1208:1213 Lys546 1208:1213 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 6 70 theme cationic 1180:1187 arg1 Arg210 1197:1202 Arg210 1197:1202 Arg210 1197:1202 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 6 70 theme cationic 1180:1187 arg1 groups 1189:1194 two cationic groups 1176:1194 two cationic groups 1176:1194 Distinct differences are seen in the interactions made by two cationic groups, Arg210 and Lys546, behind the iron site, and these may influence the stability of the two metal sites. 15299793 7 71 theme interactions 1338:1349 arg1 Analysis 1300:1307 Analysis 1300:1307 Analysis of interdomain and interlobe interactions 1300:1349 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 9 72 theme glycan 1799:1804 arg1 residues 1776:1783 two sugar residues 1766:1783 two sugar residues of the N-lobe glycan 1766:1804 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 9 72 theme glycan 1799:1804 arg1 glycan 1799:1804 the N-lobe glycan 1788:1804 the N-lobe glycan 1788:1804 The carbohydrate is weakly defined and has been modelled to a limited extent; two sugar residues of the N-lobe glycan and six of the C-lobe glycan. 15299793 5 73 theme ion 993:995 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 73 theme ion 993:995 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 73 theme ion 993:995 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 73 theme ion 993:995 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 73 theme ion 993:995 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 7 74 theme lobe 1476:1479 arg1 movements 1481:1489 lobe movements 1476:1489 lobe movements 1476:1489 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 2 75 theme 469 427:429 arg1 molecules 439:447 469 solvent molecules 427:447 469 solvent molecules 427:447 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 5 76 theme asymmetric 947:956 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 76 theme asymmetric 947:956 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 76 theme asymmetric 947:956 arg1 differences 913:923 the only apparent differences 895:923 the only apparent differences 895:923 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 76 theme asymmetric 947:956 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 76 theme asymmetric 947:956 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 1 77 theme three-dimensional 73:89 arg1 structure 91:99 The three-dimensional structure 69:99 The three-dimensional structure of the diferric form of human lactoferrin 69:141 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 5 78 theme C-lobe 1105:1110 arg1 site 1112:1115 the C-lobe site 1101:1115 the C-lobe site 1101:1115 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 0 79 theme diferric 19:26 arg1 lactoferrin 28:38 human diferric lactoferrin 13:38 human diferric lactoferrin 13:38 Structure of human diferric lactoferrin refined at 2.2 A resolution. 15299793 1 80 theme diffractometer 245:258 arg1 set 265:267 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 synchrotron data combined with a lower resolution (3.2 A) diffractometer data set 187:267 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 7 81 theme molecule 1440:1447 arg1 flexibility 1421:1431 the known flexibility 1411:1431 the known flexibility of the molecule 1411:1447 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 2 82 theme 2Fe 398:400 arg1 ions 421:424 2Fe(3+) and 2CO(3)(2-) ions 398:424 2Fe(3+) and 2CO(3)(2-) ions 398:424 Following restrained least-squares refinement and model rebuilding the final model comprises 5330 protein atoms (691 residues), 2Fe(3+) and 2CO(3)(2-) ions, 469 solvent molecules and 98 carbohydrate atoms (eight sugar residues). 15299793 5 83 theme longer 1026:1031 arg1 binding 968:974 a slightly more asymmetric bidentate binding 931:974 a slightly more asymmetric bidentate binding of the carbonate ion to the metal 931:1008 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 83 theme longer 1026:1031 arg1 bond 1038:1041 a slightly longer Fe-O bond 1015:1041 a slightly longer Fe-O bond 1015:1041 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 83 theme longer 1026:1031 arg1 sites 866:870 The two binding sites 850:870 The two binding sites 850:870 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 83 theme longer 1026:1031 arg1 similar 886:892 similar 886:892 similar 886:892 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 7 84 theme interdomain 1312:1322 arg1 interactions 1338:1349 interdomain and interlobe interactions 1312:1349 interactions 1338:1349 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 7 85 with consistent 1395:1404 arg1 flexibility 1421:1431 the known flexibility 1411:1431 the known flexibility of the molecule 1411:1447 Analysis of interdomain and interlobe interactions shows that these are few in number which is consistent with the known flexibility of the molecule with respect to domain and lobe movements. 15299793 3 86 from reflections 682:692 arg1 range 712:716 the resolution range 8.0-2.2 A 697:726 the resolution range 8.0-2.2 A 697:726 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 3 87 theme bond 566:569 arg1 lengths 571:577 bond lengths 566:577 bond lengths 566:577 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 3 88 theme angle 595:599 arg1 distances 607:615 angle (1-3) distances 595:615 angle (1-3) distances 595:615 Root-mean-square deviations from standard geometry are 0.015 A for bond lengths and 0.038 A for angle (1-3) distances, and the final crystallographic R-factor is 0.179 for all 39 113 reflections in the resolution range 8.0-2.2 A. 15299793 1 89 theme lower 220:224 arg1 resolution 226:235 a lower resolution 218:235 a lower resolution (3.2 A) 218:243 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 1 89 theme lower 220:224 arg1 A 242:242 3.2 A 238:242 3.2 A 238:242 The three-dimensional structure of the diferric form of human lactoferrin has been refined at 2.2 A resolution, using synchrotron data combined with a lower resolution (3.2 A) diffractometer data set. 15299793 5 90 theme ligands 1061:1067 arg1 one 1046:1048 one 1046:1048 one 1046:1048 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299793 5 90 theme ligands 1061:1067 arg1 ligands 1061:1067 the Tyr ligands 1053:1067 the Tyr ligands 1053:1067 The two binding sites are extremely similar, the only apparent differences being a slightly more asymmetric bidentate binding of the carbonate ion to the metal, and a slightly longer Fe-O bond to one of the Tyr ligands, in the N-lobe site relative to the C-lobe site. 15299444 4 0 theme 5314 848:851 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 0 theme 5314 848:851 arg1 ion 933:935 one oxalate ion 921:935 one oxalate ion 921:935 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 0 theme 5314 848:851 arg1 residue 974:980 one sugar residue 964:980 one sugar residue 964:980 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 0 theme 5314 848:851 arg1 ion 916:918 one bicarbonate ion 900:918 one bicarbonate ion 900:918 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 0 theme 5314 848:851 arg1 residues 872:879 691 residues 868:879 691 residues 868:879 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 0 theme 5314 848:851 arg1 molecules 950:958 325 solvent molecules 938:958 325 solvent molecules 938:958 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 0 theme 5314 848:851 arg1 ions 894:897 two Cu(2+) ions 883:897 two Cu(2+) ions 883:897 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 6 1 theme 1,2-bidentate 1132:1144 arg1 fashion 1146:1152 a 1,2-bidentate fashion 1130:1152 a 1,2-bidentate fashion 1130:1152 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 5 2 theme 134 1032:1034 arg1 reflections 1036:1046 46 134 reflections 1029:1046 46 134 reflections in the range 8.0 1029:1063 The crystallographic R factor of 0.193 is for 46 134 reflections in the range 8.0 to 2.0 A resolution. 15299444 1 3 theme X-ray 217:221 arg1 data 235:238 X-ray diffraction data 217:238 X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution 217:359 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 3 4 theme restrained 728:737 arg1 methods 753:759 restrained least-squares methods 728:759 restrained least-squares methods 728:759 The structure was then refined crystallographically by restrained least-squares methods. 15299444 1 5 theme diffraction 223:233 arg1 data 235:238 X-ray diffraction data 217:238 X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution 217:359 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 5 6 from reflections 1036:1046 arg1 range 1055:1059 the range 8.0 1051:1063 the range 8.0 1051:1063 The crystallographic R factor of 0.193 is for 46 134 reflections in the range 8.0 to 2.0 A resolution. 15299444 3 7 theme least-squares 739:751 arg1 methods 753:759 restrained least-squares methods 728:759 restrained least-squares methods 728:759 The structure was then refined crystallographically by restrained least-squares methods. 15299444 5 8 theme R 1004:1004 arg1 factor 1006:1011 The crystallographic R factor 983:1011 The crystallographic R factor of 0.193 983:1020 The crystallographic R factor of 0.193 is for 46 134 reflections in the range 8.0 to 2.0 A resolution. 15299444 4 9 theme solvent 942:948 arg1 molecules 950:958 325 solvent molecules 938:958 325 solvent molecules 938:958 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 9 theme solvent 942:948 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 1 10 theme synchrotron 322:332 arg1 source 334:339 a synchrotron source 320:339 a synchrotron source to 2.0 A resolution 320:359 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 4 11 contain contains 839:846 arg2 residues 872:879 691 residues 868:879 691 residues 868:879 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg2 ion 916:918 one bicarbonate ion 900:918 one bicarbonate ion 900:918 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg2 ions 894:897 two Cu(2+) ions 883:897 two Cu(2+) ions 883:897 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg2 molecules 950:958 325 solvent molecules 938:958 325 solvent molecules 938:958 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg1 model 772:776 The final model 762:776 The final model 762:776 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg2 ion 933:935 one oxalate ion 921:935 one oxalate ion 921:935 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg2 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 11 contain contains 839:846 arg2 residue 974:980 one sugar residue 964:980 one sugar residue 964:980 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 12 theme oxalate 925:931 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 12 theme oxalate 925:931 arg1 ion 933:935 one oxalate ion 921:935 one oxalate ion 921:935 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 13 from deviation 799:807 arg1 distances 817:825 bond distances 812:825 bond distances 812:825 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 2 14 from carbonate 538:546 arg1 site 574:577 the C-terminal binding site 551:577 the C-terminal binding site 551:577 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 2 15 theme Difference 362:371 arg1 maps 390:393 Difference electron-density maps 362:393 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, 362:497 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 4 16 theme bond 812:815 arg1 distances 817:825 bond distances 812:825 bond distances 812:825 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 2 17 theme dicupric 433:440 arg1 lactoferrin 442:452 both dicupric lactoferrin 428:452 both dicupric lactoferrin 428:452 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 5 18 theme A 1072:1072 arg1 resolution 1074:1083 2.0 A resolution 1068:1083 2.0 A resolution 1068:1083 The crystallographic R factor of 0.193 is for 46 134 reflections in the range 8.0 to 2.0 A resolution. 15299444 6 19 theme anion 1181:1185 arg1 bulk 1169:1172 the added bulk 1159:1172 the added bulk of the anion 1159:1185 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 1 20 theme oxLf 164:167 arg1 structure 106:114 The three-dimensional structure 84:114 The three-dimensional structure 84:114 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 6 21 theme residues 1267:1274 arg1 chains 1228:1233 the side chains 1219:1233 the side chains of nearby arginine and tyrosine residues 1219:1274 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 4 22 theme r.m.s. 792:797 arg1 0.017 830:834 0.017 830:834 0.017 830:834 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 22 theme r.m.s. 792:797 arg1 deviation 799:807 the r.m.s. deviation 788:807 the r.m.s. deviation in bond distances 788:825 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 2 23 theme binding 566:572 arg1 site 574:577 the C-terminal binding site 551:577 the C-terminal binding site 551:577 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 1 24 theme three-dimensional 88:104 arg1 structure 106:114 The three-dimensional structure 84:114 The three-dimensional structure 84:114 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 4 25 theme bicarbonate 904:914 arg1 ion 916:918 one bicarbonate ion 900:918 one bicarbonate ion 900:918 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 25 theme bicarbonate 904:914 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 7 26 theme other 1280:1284 arg1 alterations 1292:1302 No other major alterations 1277:1302 No other major alterations in the molecule 1277:1318 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 2 27 theme C-terminal 555:564 arg1 site 574:577 the C-terminal binding site 551:577 the C-terminal binding site 551:577 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 2 28 theme significant 625:635 arg1 differences 637:647 no significant differences 622:647 no significant differences in the N-terminal site 622:670 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 0 29 theme copper- 13:19 arg1 lactoferrin 51:61 copper- and oxalate-substituted human lactoferrin 13:61 copper- and oxalate-substituted human lactoferrin 13:61 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 15299444 7 30 from alterations 1292:1302 arg1 molecule 1311:1318 the molecule 1307:1318 the molecule 1307:1318 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 6 31 theme added 1163:1167 arg1 bulk 1169:1172 the added bulk 1159:1172 the added bulk of the anion 1159:1185 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 6 32 theme arginine 1245:1252 arg1 residues 1267:1274 nearby arginine and tyrosine residues 1238:1274 nearby arginine and tyrosine residues 1238:1274 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 6 33 theme tyrosine 1258:1265 arg1 residues 1267:1274 nearby arginine and tyrosine residues 1238:1274 nearby arginine and tyrosine residues 1238:1274 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 0 34 theme human 45:49 arg1 lactoferrin 51:61 copper- and oxalate-substituted human lactoferrin 13:61 copper- and oxalate-substituted human lactoferrin 13:61 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 15299444 2 35 theme diferric 468:475 arg1 lactoferrin 477:487 diferric lactoferrin 468:487 diferric lactoferrin 468:487 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 0 36 from resolution 72:81 arg1 Structure 0:8 Structure 0:8 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 0:82 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 15299444 6 37 theme nearby 1238:1243 arg1 residues 1267:1274 nearby arginine and tyrosine residues 1238:1274 nearby arginine and tyrosine residues 1238:1274 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 0 38 theme oxalate-substituted 25:43 arg1 lactoferrin 51:61 copper- and oxalate-substituted human lactoferrin 13:61 copper- and oxalate-substituted human lactoferrin 13:61 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 15299444 4 39 theme protein 853:859 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 39 theme protein 853:859 arg1 ion 933:935 one oxalate ion 921:935 one oxalate ion 921:935 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 39 theme protein 853:859 arg1 residue 974:980 one sugar residue 964:980 one sugar residue 964:980 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 39 theme protein 853:859 arg1 ion 916:918 one bicarbonate ion 900:918 one bicarbonate ion 900:918 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 39 theme protein 853:859 arg1 residues 872:879 691 residues 868:879 691 residues 868:879 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 39 theme protein 853:859 arg1 molecules 950:958 325 solvent molecules 938:958 325 solvent molecules 938:958 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 39 theme protein 853:859 arg1 ions 894:897 two Cu(2+) ions 883:897 two Cu(2+) ions 883:897 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 5 40 theme crystallographic 987:1002 arg1 factor 1006:1011 The crystallographic R factor 983:1011 The crystallographic R factor of 0.193 983:1020 The crystallographic R factor of 0.193 is for 46 134 reflections in the range 8.0 to 2.0 A resolution. 15299444 6 41 theme oxalate 1090:1096 arg1 ion 1098:1100 The oxalate ion 1086:1100 The oxalate ion 1086:1100 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 4 42 theme sugar 968:972 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 42 theme sugar 968:972 arg1 residue 974:980 one sugar residue 964:980 one sugar residue 964:980 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 2 43 theme electron-density 373:388 arg1 maps 390:393 Difference electron-density maps 362:393 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, 362:497 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 7 44 theme overall 1341:1347 arg1 structure 1357:1365 the overall protein structure 1337:1365 the overall protein structure 1337:1365 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 7 44 theme overall 1341:1347 arg1 same 1377:1380 same 1377:1380 same 1377:1380 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 7 45 theme protein 1349:1355 arg1 structure 1357:1365 the overall protein structure 1337:1365 the overall protein structure 1337:1365 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 7 45 theme protein 1349:1355 arg1 same 1377:1380 same 1377:1380 same 1377:1380 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 4 46 theme final 766:770 arg1 model 772:776 The final model 762:776 The final model 762:776 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 7 47 dep observed 1327:1334 arg1 structure 1357:1365 the overall protein structure 1337:1365 the overall protein structure 1337:1365 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 7 47 dep observed 1327:1334 arg1 same 1377:1380 same 1377:1380 same 1377:1380 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 1 48 theme human 119:123 arg1 lactoferrin 146:156 human dicupric monooxalate lactoferrin 119:156 human dicupric monooxalate lactoferrin 119:156 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 0 49 theme lactoferrin 51:61 arg1 Structure 0:8 Structure 0:8 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 0:82 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 15299444 2 50 dep Cu 414:415 arg1 2 417:417 2 417:417 2 417:417 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 2 50 dep Cu 414:415 arg1 oxLf 419:422 oxLf 419:422 oxLf 419:422 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 1 51 theme dicupric 125:132 arg1 lactoferrin 146:156 human dicupric monooxalate lactoferrin 119:156 human dicupric monooxalate lactoferrin 119:156 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 4 52 dep 0.017 830:834 arg1 A 836:836 A 836:836 A 836:836 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 1 53 theme monooxalate 134:144 arg1 lactoferrin 146:156 human dicupric monooxalate lactoferrin 119:156 human dicupric monooxalate lactoferrin 119:156 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 2 54 theme N-terminal 656:665 arg1 site 667:670 the N-terminal site 652:670 the N-terminal site 652:670 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 2 55 dep Lf 607:608 arg1 2 605:605 2 605:605 2 605:605 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 1 56 theme lactoferrin 146:156 arg1 structure 106:114 The three-dimensional structure 84:114 The three-dimensional structure 84:114 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 6 57 theme side 1223:1226 arg1 chains 1228:1233 the side chains 1219:1233 the side chains of nearby arginine and tyrosine residues 1219:1274 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 5 58 theme 0.193 1016:1020 arg1 factor 1006:1011 The crystallographic R factor 983:1011 The crystallographic R factor of 0.193 983:1020 The crystallographic R factor of 0.193 is for 46 134 reflections in the range 8.0 to 2.0 A resolution. 15299444 0 59 theme A 70:70 arg1 resolution 72:81 2.0 A resolution 66:81 2.0 A resolution 66:81 Structure of copper- and oxalate-substituted human lactoferrin at 2.0 A resolution. 15299444 4 60 theme Cu 887:888 arg1 atoms 861:865 5314 protein atoms 848:865 5314 protein atoms (691 residues) 848:880 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 4 60 theme Cu 887:888 arg1 ions 894:897 two Cu(2+) ions 883:897 two Cu(2+) ions 883:897 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 2 61 from differences 637:647 arg1 site 667:670 the N-terminal site 652:670 the N-terminal site 652:670 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 15299444 6 62 theme chains 1228:1233 arg1 rearrangement 1202:1214 the rearrangement 1198:1214 the rearrangement of the side chains of nearby arginine and tyrosine residues 1198:1274 The oxalate ion is coordinated to copper in a 1,2-bidentate fashion, and the added bulk of the anion results in the rearrangement of the side chains of nearby arginine and tyrosine residues. 15299444 4 63 from 0.017 830:834 arg1 model 772:776 The final model 762:776 The final model 762:776 The final model, in which the r.m.s. deviation in bond distances is 0.017 A, contains 5314 protein atoms (691 residues), two Cu(2+) ions, one bicarbonate ion, one oxalate ion, 325 solvent molecules and one sugar residue. 15299444 7 64 theme major 1286:1290 arg1 alterations 1292:1302 No other major alterations 1277:1302 No other major alterations in the molecule 1277:1318 No other major alterations in the molecule can be observed, the overall protein structure being the same as that for Cu(2)Lf and Fe(2)Lf. 15299444 1 65 theme oscillation 293:303 arg1 photography 305:315 oscillation photography 293:315 oscillation photography 293:315 The three-dimensional structure of human dicupric monooxalate lactoferrin, Cu(2)oxLf, has been determined to 2.0 A resolution, using X-ray diffraction data collected by diffractometry to 2.5 A resolution, and oscillation photography on a synchrotron source to 2.0 A resolution. 15299444 2 66 theme Cu 602:603 arg1 Lf 607:608 Cu(2)Lf 602:608 Cu(2)Lf 602:608 Difference electron-density maps calculated between Cu(2)oxLf and both dicupric lactoferrin, Cu(2)Lf, and diferric lactoferrin, Fe(2)Lf, showed that the oxalate had replaced a carbonate in the C-terminal binding site, and that, relative to Cu(2)Lf, there were no significant differences in the N-terminal site. 19805286 9 0 theme membrane-associated 1500:1518 arg1 alpha-CAs 1520:1528 the other membrane-associated alpha-CAs 1490:1528 the other membrane-associated alpha-CAs 1490:1528 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 2 1 theme human 307:311 arg1 cancers 313:319 several human cancers 299:319 several human cancers 299:319 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 6 2 theme catalytic 846:854 arg1 domains 856:862 two catalytic domains 842:862 two catalytic domains of CA IX 842:871 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 6 2 theme catalytic 846:854 arg1 IX 870:871 CA IX 867:871 CA IX 867:871 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 4 3 theme X-ray 549:553 arg1 structure 555:563 the X-ray structure 545:563 the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide 545:678 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 9 4 theme structural 1440:1449 arg1 differences 1451:1461 the structural differences 1436:1461 the structural differences observed between CA IX and the other membrane-associated alpha-CAs 1436:1528 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 3 5 theme cell 507:510 arg1 control 521:527 cell adhesion control 507:527 cell adhesion control 507:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 3 5 theme cell 507:510 arg1 mechanisms 477:486 at least two mechanisms 464:486 at least two mechanisms: pH regulation and cell adhesion control 464:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 9 6 dep differences 1451:1461 arg1 the 1423:1425 the 1423:1425 the 1423:1425 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 9 6 dep differences 1451:1461 arg1 basis 1427:1431 basis 1427:1431 basis 1427:1431 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 7 7 theme PG 1005:1006 arg1 domains 1008:1014 the PG domains 1001:1014 the PG domains 1001:1014 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 7 theme PG 1005:1006 arg1 located 1020:1026 located 1020:1026 located 1020:1026 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 1 8 theme alpha-CA 168:175 arg1 family 184:189 the alpha-CA enzyme family 164:189 the alpha-CA enzyme family 164:189 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 9 9 theme drug 1566:1569 arg1 design 1571:1576 the rational drug design 1553:1576 the rational drug design of isozyme-specific CA inhibitors 1553:1610 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 2 10 theme tumor 258:262 arg1 hypoxia 264:270 tumor hypoxia 258:270 tumor hypoxia 258:270 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 1 11 theme family 184:189 arg1 member 154:159 a plasma membrane-associated member 125:159 a plasma membrane-associated member 125:159 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 1 11 theme family 184:189 arg1 IX 119:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 4 12 theme IX 595:596 arg1 domain 582:587 the catalytic domain 568:587 the catalytic domain of CA IX 568:596 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 4 12 theme IX 595:596 arg1 IX 595:596 CA IX 592:596 CA IX 592:596 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 7 13 theme site 985:988 arg1 clefts 990:995 The active site clefts 974:995 The active site clefts 974:995 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 13 theme site 985:988 arg1 located 1020:1026 located 1020:1026 located 1020:1026 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 5 14 theme protein 792:798 arg1 structure 811:819 the protein quaternary structure 788:819 the protein quaternary structure 788:819 The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. 19805286 9 15 theme CA 1598:1599 arg1 inhibitors 1601:1610 isozyme-specific CA inhibitors 1581:1610 isozyme-specific CA inhibitors 1581:1610 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 8 16 theme activity 1335:1342 arg1 profile 1310:1316 the pH profile 1303:1316 the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons 1303:1417 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 6 17 theme IX 870:871 arg1 domains 856:862 two catalytic domains 842:862 two catalytic domains of CA IX 842:871 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 6 17 theme IX 870:871 arg1 IX 870:871 CA IX 867:871 CA IX 867:871 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 3 18 dep mechanisms 477:486 arg1 control 521:527 cell adhesion control 507:527 cell adhesion control 507:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 3 18 dep mechanisms 477:486 arg1 mechanisms 477:486 at least two mechanisms 464:486 at least two mechanisms: pH regulation and cell adhesion control 464:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 3 18 dep mechanisms 477:486 arg1 regulation 492:501 pH regulation 489:501 pH regulation 489:501 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 7 19 theme protein 1121:1127 arg1 anchoring 1129:1137 protein anchoring 1121:1137 protein anchoring to the cell membrane 1121:1158 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 0 20 theme carbonic 72:79 arg1 anhydrase 81:89 the tumor-associated human carbonic anhydrase IX 45:92 the tumor-associated human carbonic anhydrase IX 45:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 1 21 theme membrane-associated 134:152 arg1 member 154:159 a plasma membrane-associated member 125:159 a plasma membrane-associated member 125:159 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 1 21 theme membrane-associated 134:152 arg1 IX 119:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 8 22 theme 2 1379:1379 arg1 reaction 1366:1373 the physiological reaction 1348:1373 the physiological reaction 1348:1373 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 8 22 theme 2 1379:1379 arg1 hydration 1382:1390 CO(2) hydration 1376:1390 CO(2) hydration to bicarbonate and protons 1376:1417 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 7 23 located located 1020:1026 arg2 clefts 990:995 The active site clefts 974:995 The active site clefts 974:995 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 23 located located 1020:1026 arg2 domains 1008:1014 the PG domains 1001:1014 the PG domains 1001:1014 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 23 located located 1020:1026 arg2 located 1020:1026 located 1020:1026 located 1020:1026 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 23 located located 1020:1026 arg1 face 1035:1038 one face 1031:1038 one face of the dimer 1031:1051 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 24 theme opposite 1093:1100 arg1 face 1102:1105 the opposite face 1089:1105 the opposite face to facilitate protein anchoring to the cell membrane 1089:1158 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 9 25 theme enzyme 1656:1661 arg1 inhibition 1637:1646 inhibition 1637:1646 inhibition of this enzyme 1637:1661 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 0 26 theme domain 35:40 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 0:93 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 4 27 dep classical 616:624 arg1 used 638:641 used 638:641 used 638:641 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 9 28 theme antitumor 1669:1677 arg1 activity 1679:1686 antitumor activity 1669:1686 antitumor activity 1669:1686 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 0 29 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 0:93 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 3 30 theme aberrant 325:332 arg1 increase 334:341 An aberrant increase 322:341 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas 322:425 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 3 31 theme carcinomas 416:425 arg1 development 393:403 development 393:403 development of various carcinomas 393:425 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 5 32 theme CA 771:772 arg1 isozymes 774:781 the other CA isozymes 761:781 the other CA isozymes 761:781 The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. 19805286 1 33 theme anhydrase 104:112 arg1 member 154:159 a plasma membrane-associated member 125:159 a plasma membrane-associated member 125:159 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 1 33 theme anhydrase 104:112 arg1 IX 119:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 0 34 theme tumor-associated 49:64 arg1 anhydrase 81:89 the tumor-associated human carbonic anhydrase IX 45:92 the tumor-associated human carbonic anhydrase IX 45:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 0 35 theme catalytic 25:33 arg1 domain 35:40 the catalytic domain 21:40 the catalytic domain of the tumor-associated human carbonic anhydrase IX 21:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 0 35 theme catalytic 25:33 arg1 anhydrase 81:89 the tumor-associated human carbonic anhydrase IX 45:92 the tumor-associated human carbonic anhydrase IX 45:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 6 36 theme disulfide 958:966 arg1 bond 968:971 an intermolecular disulfide bond 940:971 an intermolecular disulfide bond 940:971 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 3 37 theme IX 349:350 arg1 expression 352:361 CA IX expression 346:361 CA IX expression in chronic hypoxia 346:380 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 5 38 theme typical 705:711 arg1 fold 722:725 a typical alpha-CA fold 703:725 a typical alpha-CA fold 703:725 The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. 19805286 2 39 theme several 299:305 arg1 cancers 313:319 several human cancers 299:319 several human cancers 299:319 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 9 40 theme CA 1480:1481 arg1 IX 1483:1484 CA IX 1480:1484 CA IX 1480:1484 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 8 41 theme pH 1307:1308 arg1 profile 1310:1316 the pH profile 1303:1316 the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons 1303:1417 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 7 42 located located 1078:1084 arg2 located 1078:1084 located 1078:1084 located 1078:1084 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 42 located located 1078:1084 arg2 C-termini 1064:1072 the C-termini 1060:1072 the C-termini 1060:1072 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 42 located located 1078:1084 arg1 face 1102:1105 the opposite face 1089:1105 the opposite face to facilitate protein anchoring to the cell membrane 1089:1158 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 6 43 theme CA 867:868 arg1 IX 870:871 CA IX 867:871 CA IX 867:871 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 9 44 theme other 1494:1498 arg1 alpha-CAs 1520:1528 the other membrane-associated alpha-CAs 1490:1528 the other membrane-associated alpha-CAs 1490:1528 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 7 45 theme cell 1146:1149 arg1 membrane 1151:1158 the cell membrane 1142:1158 the cell membrane 1142:1158 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 5 46 theme alpha-CA 713:720 arg1 fold 722:725 a typical alpha-CA fold 703:725 a typical alpha-CA fold 703:725 The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. 19805286 1 47 theme tumor 219:223 arg1 acidification 225:237 solid tumor acidification 213:237 solid tumor acidification 213:237 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 2 48 from factor 289:294 arg1 cancers 313:319 several human cancers 299:319 several human cancers 299:319 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 9 49 theme further 1531:1537 arg1 prospects 1539:1547 further prospects 1531:1547 further prospects for the rational drug design of isozyme-specific CA inhibitors 1531:1610 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 3 50 theme pH 489:490 arg1 mechanisms 477:486 at least two mechanisms 464:486 at least two mechanisms: pH regulation and cell adhesion control 464:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 3 50 theme pH 489:490 arg1 regulation 492:501 pH regulation 489:501 pH regulation 489:501 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 5 51 theme quaternary 800:809 arg1 structure 811:819 the protein quaternary structure 788:819 the protein quaternary structure 788:819 The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. 19805286 4 52 from structure 555:563 arg1 complex 601:607 complex 601:607 complex with a classical, clinically used sulfonamide inhibitor, acetazolamide 601:678 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 9 53 theme rational 1557:1564 arg1 design 1571:1576 the rational drug design 1553:1576 the rational drug design of isozyme-specific CA inhibitors 1553:1610 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 3 54 theme adhesion 512:519 arg1 control 521:527 cell adhesion control 507:527 cell adhesion control 507:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 3 54 theme adhesion 512:519 arg1 mechanisms 477:486 at least two mechanisms 464:486 at least two mechanisms: pH regulation and cell adhesion control 464:527 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 1 55 theme enzyme 177:182 arg1 family 184:189 the alpha-CA enzyme family 164:189 the alpha-CA enzyme family 164:189 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 8 56 theme catalytic 1325:1333 arg1 activity 1335:1342 the catalytic activity 1321:1342 the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons 1321:1417 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 4 57 theme domain 582:587 arg1 structure 555:563 the X-ray structure 545:563 the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide 545:678 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 7 58 theme active 978:983 arg1 clefts 990:995 The active site clefts 974:995 The active site clefts 974:995 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 7 58 theme active 978:983 arg1 located 1020:1026 located 1020:1026 located 1020:1026 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 9 59 theme isozyme-specific 1581:1596 arg1 inhibitors 1601:1610 isozyme-specific CA inhibitors 1581:1610 isozyme-specific CA inhibitors 1581:1610 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 4 60 theme CA 592:593 arg1 IX 595:596 CA IX 592:596 CA IX 592:596 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 9 61 theme inhibitors 1601:1610 arg1 design 1571:1576 the rational drug design 1553:1576 the rational drug design of isozyme-specific CA inhibitors 1553:1610 On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. 19805286 8 62 theme physiological 1352:1364 arg1 reaction 1366:1373 the physiological reaction 1348:1373 the physiological reaction 1348:1373 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 8 62 theme physiological 1352:1364 arg1 hydration 1382:1390 CO(2) hydration 1376:1390 CO(2) hydration to bicarbonate and protons 1376:1417 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 4 63 theme sulfonamide 643:653 arg1 inhibitor 655:663 a classical, clinically used sulfonamide inhibitor 614:663 a classical, clinically used sulfonamide inhibitor 614:663 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 4 63 theme sulfonamide 643:653 arg1 acetazolamide 666:678 acetazolamide 666:678 acetazolamide 666:678 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 2 64 theme hypoxia 264:270 arg1 marker 248:253 a marker 246:253 a marker of tumor hypoxia 246:270 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 2 64 theme hypoxia 264:270 arg1 factor 289:294 a prognostic factor 276:294 a prognostic factor in several human cancers 276:319 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 2 64 theme hypoxia 264:270 arg1 It 240:241 It 240:241 It 240:241 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 6 65 theme intermolecular 943:956 arg1 bond 968:971 an intermolecular disulfide bond 940:971 an intermolecular disulfide bond 940:971 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 8 66 theme three-dimensional 1187:1203 arg1 structure 1205:1213 the three-dimensional structure 1183:1213 the three-dimensional structure 1183:1213 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 0 67 theme human 66:70 arg1 anhydrase 81:89 the tumor-associated human carbonic anhydrase IX 45:92 the tumor-associated human carbonic anhydrase IX 45:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 2 68 from marker 248:253 arg1 cancers 313:319 several human cancers 299:319 several human cancers 299:319 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 4 69 theme classical 616:624 arg1 inhibitor 655:663 a classical, clinically used sulfonamide inhibitor 614:663 a classical, clinically used sulfonamide inhibitor 614:663 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 4 69 theme classical 616:624 arg1 acetazolamide 666:678 acetazolamide 666:678 acetazolamide 666:678 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 0 70 theme anhydrase 81:89 arg1 domain 35:40 the catalytic domain 21:40 the catalytic domain of the tumor-associated human carbonic anhydrase IX 21:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 0 70 theme anhydrase 81:89 arg1 anhydrase 81:89 the tumor-associated human carbonic anhydrase IX 45:92 the tumor-associated human carbonic anhydrase IX 45:92 Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. 19805286 3 71 from expression 352:361 arg1 hypoxia 374:380 chronic hypoxia 366:380 chronic hypoxia 366:380 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 1 72 theme plasma 127:132 arg1 member 154:159 a plasma membrane-associated member 125:159 a plasma membrane-associated member 125:159 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 1 72 theme plasma 127:132 arg1 IX 119:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 8 73 theme physiological 1223:1235 arg1 role 1237:1240 the physiological role 1219:1240 the physiological role of the enzyme 1219:1254 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 3 74 from increase 334:341 arg1 expression 352:361 CA IX expression 346:361 CA IX expression in chronic hypoxia 346:380 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 8 75 theme enzyme 1249:1254 arg1 structure 1205:1213 the three-dimensional structure 1183:1213 the three-dimensional structure 1183:1213 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 8 75 theme enzyme 1249:1254 arg1 role 1237:1240 the physiological role 1219:1240 the physiological role of the enzyme 1219:1254 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 3 76 theme CA 346:347 arg1 expression 352:361 CA IX expression 346:361 CA IX expression in chronic hypoxia 346:380 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 5 77 theme other 765:769 arg1 isozymes 774:781 the other CA isozymes 761:781 the other CA isozymes 761:781 The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. 19805286 1 78 theme Carbonic 95:102 arg1 CA 115:116 CA 115:116 CA 115:116 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 1 78 theme Carbonic 95:102 arg1 anhydrase 104:112 Carbonic anhydrase 95:112 Carbonic anhydrase (CA) IX 95:120 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 19805286 7 79 theme dimer 1047:1051 arg1 face 1035:1038 one face 1031:1038 one face of the dimer 1031:1051 The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. 19805286 6 80 theme bond 968:971 arg1 formation 927:935 the formation 923:935 the formation of an intermolecular disulfide bond 923:971 Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. 19805286 3 81 theme chronic 366:372 arg1 hypoxia 374:380 chronic hypoxia 366:380 chronic hypoxia 366:380 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 4 82 with complex 601:607 arg1 inhibitor 655:663 a classical, clinically used sulfonamide inhibitor 614:663 a classical, clinically used sulfonamide inhibitor 614:663 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 4 82 with complex 601:607 arg1 acetazolamide 666:678 acetazolamide 666:678 acetazolamide 666:678 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 3 83 theme various 408:414 arg1 carcinomas 416:425 various carcinomas 408:425 various carcinomas 408:425 An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. 19805286 2 84 theme prognostic 278:287 arg1 factor 289:294 a prognostic factor 276:294 a prognostic factor in several human cancers 276:319 It is a marker of tumor hypoxia and a prognostic factor in several human cancers. 19805286 4 85 theme catalytic 572:580 arg1 domain 582:587 the catalytic domain 568:587 the catalytic domain of CA IX 568:596 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 4 85 theme catalytic 572:580 arg1 IX 595:596 CA IX 592:596 CA IX 592:596 Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. 19805286 8 86 theme profile 1310:1316 arg1 measurement 1288:1298 the measurement 1284:1298 the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons 1284:1417 A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. 19805286 1 87 theme solid 213:217 arg1 acidification 225:237 solid tumor acidification 213:237 solid tumor acidification 213:237 Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. 25752612 4 0 theme similar 878:884 arg1 mechanism 868:876 a catalytic mechanism 856:876 a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 856:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 5 1 contain possesses 1026:1034 arg2 activity 1075:1082 previously unrecognized aminopeptidase activity 1036:1082 previously unrecognized aminopeptidase activity 1036:1082 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 5 1 contain possesses 1026:1034 arg1 product 1018:1024 the NAALADL1 gene product 1000:1024 the NAALADL1 gene product 1000:1024 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 5 1 contain possesses 1026:1034 arg2 activity 1117:1124 no carboxy- or endopeptidase activity 1088:1124 no carboxy- or endopeptidase activity 1088:1124 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 1 2 link α-linked 105:112 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 2 link α-linked 105:112 arg1 L 157:157 NAALADase L 147:157 NAALADase L 147:157 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 2 link α-linked 105:112 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 6 3 theme enzyme 1249:1254 arg1 inhibitor 1232:1240 a potent inhibitor 1223:1240 a potent inhibitor of the enzyme 1223:1254 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 6 3 theme enzyme 1249:1254 arg1 findings 1133:1140 These findings 1127:1140 These findings 1127:1140 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 8 4 from stages 1605:1610 arg1 system 1679:1684 the human digestive system 1659:1684 the human digestive system 1659:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 3 5 theme protein 534:540 arg1 product 542:548 the protein product 530:548 the protein product of the human NAALADL1 gene 530:575 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 9 6 theme new 1724:1726 arg1 name 1728:1731 a new name 1722:1731 a new name for this enzyme: human ileal aminopeptidase (HILAP) 1722:1783 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 1 7 theme NAALADL1 176:183 arg1 gene 185:188 the NAALADL1 gene 172:188 the NAALADL1 gene 172:188 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 4 8 theme catalytic 858:866 arg1 mechanism 868:876 a catalytic mechanism 856:876 a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 856:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 6 9 theme potent 1225:1230 arg1 inhibitor 1232:1240 a potent inhibitor 1223:1240 a potent inhibitor of the enzyme 1223:1254 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 6 9 theme potent 1225:1230 arg1 findings 1133:1140 These findings 1127:1140 These findings 1127:1140 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 3 10 theme human 557:561 arg1 gene 572:575 the human NAALADL1 gene 553:575 the human NAALADL1 gene 553:575 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 4 11 theme distinct 931:938 arg1 specificity 950:960 distinct substrate specificity 931:960 distinct substrate specificity 931:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 7 12 theme NAALADL1 1455:1462 arg1 transcript 1469:1478 the NAALADL1 gene transcript 1451:1478 the NAALADL1 gene transcript 1451:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 8 13 from system 1679:1684 arg1 stages 1605:1610 the final stages 1595:1610 the final stages of protein/peptide digestion and absorption in the human digestive system 1595:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 9 14 theme human 1750:1754 arg1 HILAP 1778:1782 HILAP 1778:1782 HILAP 1778:1782 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 9 14 theme human 1750:1754 arg1 aminopeptidase 1762:1775 human ileal aminopeptidase 1750:1775 a new name for this enzyme: human ileal aminopeptidase (HILAP) 1722:1783 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 7 15 from Analysis 1257:1264 arg1 levels 1323:1328 both the mRNA and protein levels 1297:1328 levels 1323:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 4 16 theme glutamate 897:905 arg1 carboxypeptidase 907:922 glutamate carboxypeptidase II 897:925 glutamate carboxypeptidase II 897:925 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 7 17 theme alternative 1427:1437 arg1 splicing 1439:1446 extensive alternative splicing 1417:1446 extensive alternative splicing of the NAALADL1 gene transcript 1417:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 3 18 theme gene 572:575 arg1 product 542:548 the protein product 530:548 the protein product of the human NAALADL1 gene 530:575 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 8 19 theme NAALADL1 1521:1528 arg1 product 1535:1541 the NAALADL1 gene product 1517:1541 the NAALADL1 gene product's primary physiological function 1517:1574 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 5 20 theme proteome-based 965:978 arg1 assay 980:984 A proteome-based assay 963:984 A proteome-based assay 963:984 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 8 21 theme primary 1545:1551 arg1 function 1567:1574 the NAALADL1 gene product's primary physiological function 1517:1574 the NAALADL1 gene product's primary physiological function 1517:1574 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 7 22 theme expression 1283:1292 arg1 Analysis 1257:1264 Analysis 1257:1264 Analysis of NAALADL1 gene expression at both the mRNA and protein levels 1257:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 23 theme NAALADL1 1269:1276 arg1 expression 1283:1292 NAALADL1 gene expression 1269:1292 NAALADL1 gene expression at both the mRNA and protein levels 1269:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 1 24 theme solid 336:340 arg1 malignancies 342:353 solid malignancies 336:353 solid malignancies 336:353 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 5 25 theme NAALADL1 1004:1011 arg1 product 1018:1024 the NAALADL1 gene product 1000:1024 the NAALADL1 gene product 1000:1024 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 7 26 theme expression 1388:1397 arg1 points 1403:1408 points 1403:1408 points toward extensive alternative splicing of the NAALADL1 gene transcript 1403:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 26 theme expression 1388:1397 arg1 site 1372:1375 the major site 1362:1375 the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript 1362:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 26 theme expression 1388:1397 arg1 expression 1388:1397 protein expression 1380:1397 protein expression 1380:1397 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 2 27 theme NAALADase 449:457 arg1 L 459:459 NAALADase L 449:459 NAALADase L 449:459 However, neither the physiological functions nor structural features of NAALADase L are known at present. 25752612 4 28 theme L 744:744 arg1 structure 752:760 the NAALADase L x-ray structure 730:760 the NAALADase L x-ray structure 730:760 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 0 29 theme Structural 0:9 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 0:90 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 1 30 theme α-linked 105:112 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 30 theme α-linked 105:112 arg1 L 157:157 NAALADase L 147:157 NAALADase L 147:157 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 30 theme α-linked 105:112 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 0 31 theme biochemical 15:25 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 0:90 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 1 32 theme dipeptidase-like 121:136 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 32 theme dipeptidase-like 121:136 arg1 L 157:157 NAALADase L 147:157 NAALADase L 147:157 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 32 theme dipeptidase-like 121:136 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 7 33 theme major 1366:1370 arg1 points 1403:1408 points 1403:1408 points toward extensive alternative splicing of the NAALADL1 gene transcript 1403:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 33 theme major 1366:1370 arg1 site 1372:1375 the major site 1362:1375 the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript 1362:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 33 theme major 1366:1370 arg1 expression 1388:1397 protein expression 1380:1397 protein expression 1380:1397 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 8 34 theme protein/peptide 1615:1629 arg1 digestion 1631:1639 protein/peptide digestion 1615:1639 protein/peptide digestion 1615:1639 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 3 35 theme biochemical 649:659 arg1 characterization 661:676 structural and biochemical characterization 634:676 structural and biochemical characterization 634:676 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 2 36 theme physiological 398:410 arg1 functions 412:420 the physiological functions 394:420 the physiological functions 394:420 However, neither the physiological functions nor structural features of NAALADase L are known at present. 25752612 5 37 theme aminopeptidase 1060:1073 arg1 activity 1075:1082 previously unrecognized aminopeptidase activity 1036:1082 previously unrecognized aminopeptidase activity 1036:1082 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 3 38 theme structural 634:643 arg1 characterization 661:676 structural and biochemical characterization 634:676 structural and biochemical characterization 634:676 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 4 39 theme zinc-dependent 819:832 arg1 metallopeptidase 834:849 a zinc-dependent metallopeptidase 817:849 a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 817:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 4 39 theme zinc-dependent 819:832 arg1 it 811:812 it 811:812 it 811:812 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 0 40 theme novel 49:53 arg1 aminopeptidase 55:68 a novel aminopeptidase 47:68 a novel aminopeptidase from human intestine 47:89 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 0 41 from characterization 27:42 arg1 intestine 81:89 human intestine 75:89 human intestine 75:89 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 7 42 theme small 1343:1347 arg1 intestine 1349:1357 the small intestine 1339:1357 the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript 1339:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 4 43 theme experimental 784:795 arg1 evidence 797:804 the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 774:960 the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 774:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 8 44 theme human 1663:1667 arg1 system 1679:1684 the human digestive system 1659:1684 the human digestive system 1659:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 1 45 theme carboxypeptidase 223:238 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 45 theme carboxypeptidase 223:238 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 7 46 theme protein 1315:1321 arg1 levels 1323:1328 both the mRNA and protein levels 1297:1328 levels 1323:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 3 47 theme profile 710:716 arg1 overexpression 601:614 heterologous overexpression 588:614 heterologous overexpression 588:614 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 3 47 theme profile 710:716 arg1 analysis 683:690 analysis 683:690 analysis of its expression profile 683:716 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 3 47 theme profile 710:716 arg1 characterization 661:676 structural and biochemical characterization 634:676 structural and biochemical characterization 634:676 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 3 47 theme profile 710:716 arg1 purification 620:631 purification 620:631 purification 620:631 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 8 48 from digestion 1631:1639 arg1 system 1679:1684 the human digestive system 1659:1684 the human digestive system 1659:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 7 49 theme mRNA 1306:1309 arg1 levels 1323:1328 both the mRNA and protein levels 1297:1328 levels 1323:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 50 theme transcript 1469:1478 arg1 splicing 1439:1446 extensive alternative splicing 1417:1446 extensive alternative splicing of the NAALADL1 gene transcript 1417:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 9 51 dep name 1728:1731 arg1 HILAP 1778:1782 HILAP 1778:1782 HILAP 1778:1782 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 9 51 dep name 1728:1731 arg1 aminopeptidase 1762:1775 human ileal aminopeptidase 1750:1775 a new name for this enzyme: human ileal aminopeptidase (HILAP) 1722:1783 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 7 52 from levels 1323:1328 arg1 expression 1283:1292 NAALADL1 gene expression 1269:1292 NAALADL1 gene expression at both the mRNA and protein levels 1269:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 52 from levels 1323:1328 arg1 Analysis 1257:1264 Analysis 1257:1264 Analysis of NAALADL1 gene expression at both the mRNA and protein levels 1257:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 8 53 from absorption 1645:1654 arg1 system 1679:1684 the human digestive system 1659:1684 the human digestive system 1659:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 3 54 theme product 542:548 arg1 characterization 510:525 a thorough characterization 499:525 a thorough characterization 499:525 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 6 55 theme bestatin 1211:1218 arg1 identification 1193:1206 identification 1193:1206 identification of bestatin 1193:1218 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 6 55 theme bestatin 1211:1218 arg1 mutagenesis 1177:1187 site-directed mutagenesis 1163:1187 site-directed mutagenesis 1163:1187 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 1 56 theme close 196:200 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 56 theme close 196:200 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 4 57 theme substrate 940:948 arg1 specificity 950:960 distinct substrate specificity 931:960 distinct substrate specificity 931:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 3 58 theme NAALADL1 563:570 arg1 gene 572:575 the human NAALADL1 gene 553:575 the human NAALADL1 gene 553:575 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 8 59 theme gene 1530:1533 arg1 product 1535:1541 the NAALADL1 gene product 1517:1541 the NAALADL1 gene product's primary physiological function 1517:1574 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 5 60 theme carboxy- 1091:1098 arg1 activity 1117:1124 no carboxy- or endopeptidase activity 1088:1124 no carboxy- or endopeptidase activity 1088:1124 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 1 61 theme NAALADase 147:155 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 61 theme NAALADase 147:155 arg1 L 157:157 NAALADase L 147:157 NAALADase L 147:157 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 7 62 theme extensive 1417:1425 arg1 splicing 1439:1446 extensive alternative splicing 1417:1446 extensive alternative splicing of the NAALADL1 gene transcript 1417:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 0 63 theme human 75:79 arg1 intestine 81:89 human intestine 75:89 human intestine 75:89 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 5 64 theme endopeptidase 1103:1115 arg1 activity 1117:1124 no carboxy- or endopeptidase activity 1088:1124 no carboxy- or endopeptidase activity 1088:1124 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 3 65 theme heterologous 588:599 arg1 overexpression 601:614 heterologous overexpression 588:614 heterologous overexpression 588:614 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 8 66 theme physiological 1553:1565 arg1 function 1567:1574 the NAALADL1 gene product's primary physiological function 1517:1574 the NAALADL1 gene product's primary physiological function 1517:1574 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 7 67 theme points 1403:1408 arg1 points 1403:1408 points 1403:1408 points toward extensive alternative splicing of the NAALADL1 gene transcript 1403:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 67 theme points 1403:1408 arg1 site 1372:1375 the major site 1362:1375 the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript 1362:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 67 theme points 1403:1408 arg1 expression 1388:1397 protein expression 1380:1397 protein expression 1380:1397 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 68 theme gene 1278:1281 arg1 expression 1283:1292 NAALADL1 gene expression 1269:1292 NAALADL1 gene expression at both the mRNA and protein levels 1269:1328 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 7 69 theme gene 1464:1467 arg1 transcript 1469:1478 the NAALADL1 gene transcript 1451:1478 the NAALADL1 gene transcript 1451:1478 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 1 70 theme malignancies 342:353 arg1 therapy 325:331 therapy 325:331 therapy 325:331 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 70 theme malignancies 342:353 arg1 imaging 313:319 imaging 313:319 imaging 313:319 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 7 71 theme protein 1380:1386 arg1 expression 1388:1397 protein expression 1380:1397 protein expression 1380:1397 Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed the small intestine as the major site of protein expression and points toward extensive alternative splicing of the NAALADL1 gene transcript. 25752612 8 72 theme final 1599:1603 arg1 stages 1605:1610 the final stages 1595:1610 the final stages of protein/peptide digestion and absorption in the human digestive system 1595:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 1 73 theme neuropathologies 359:374 arg1 therapy 325:331 therapy 325:331 therapy 325:331 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 73 theme neuropathologies 359:374 arg1 imaging 313:319 imaging 313:319 imaging 313:319 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 4 74 with metallopeptidase 834:849 arg1 mechanism 868:876 a catalytic mechanism 856:876 a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 856:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 1 75 theme N-acetylated 92:103 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 75 theme N-acetylated 92:103 arg1 L 157:157 NAALADase L 147:157 NAALADase L 147:157 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 75 theme N-acetylated 92:103 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 9 76 theme ileal 1756:1760 arg1 HILAP 1778:1782 HILAP 1778:1782 HILAP 1778:1782 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 9 76 theme ileal 1756:1760 arg1 aminopeptidase 1762:1775 human ileal aminopeptidase 1750:1775 a new name for this enzyme: human ileal aminopeptidase (HILAP) 1722:1783 Based on these results, we suggest a new name for this enzyme: human ileal aminopeptidase (HILAP). 25752612 4 77 theme NAALADase 734:742 arg1 structure 752:760 the NAALADase L x-ray structure 730:760 the NAALADase L x-ray structure 730:760 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 0 78 from intestine 81:89 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 0:90 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 0 78 from intestine 81:89 arg1 aminopeptidase 55:68 a novel aminopeptidase 47:68 a novel aminopeptidase from human intestine 47:89 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 1 79 theme acidic 114:119 arg1 protein 138:144 N-acetylated α-linked acidic dipeptidase-like protein 92:144 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L) 92:158 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 79 theme acidic 114:119 arg1 L 157:157 NAALADase L 147:157 NAALADase L 147:157 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 79 theme acidic 114:119 arg1 homolog 202:208 a close homolog 194:208 a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 194:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 2 80 theme structural 426:435 arg1 features 437:444 structural features 426:444 structural features of NAALADase L 426:459 However, neither the physiological functions nor structural features of NAALADase L are known at present. 25752612 8 81 theme digestion 1631:1639 arg1 stages 1605:1610 the final stages 1595:1610 the final stages of protein/peptide digestion and absorption in the human digestive system 1595:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 4 82 theme x-ray 746:750 arg1 structure 752:760 the NAALADase L x-ray structure 730:760 the NAALADase L x-ray structure 730:760 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 5 83 theme unrecognized 1047:1058 arg1 activity 1075:1082 previously unrecognized aminopeptidase activity 1036:1082 previously unrecognized aminopeptidase activity 1036:1082 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 8 84 theme absorption 1645:1654 arg1 stages 1605:1610 the final stages 1595:1610 the final stages of protein/peptide digestion and absorption in the human digestive system 1595:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 0 85 theme aminopeptidase 55:68 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 0:90 Structural and biochemical characterization of a novel aminopeptidase from human intestine. 25752612 2 86 theme L 459:459 arg1 features 437:444 structural features 426:444 structural features of NAALADase L 426:459 However, neither the physiological functions nor structural features of NAALADase L are known at present. 25752612 5 87 theme gene 1013:1016 arg1 product 1018:1024 the NAALADL1 gene product 1000:1024 the NAALADL1 gene product 1000:1024 A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. 25752612 8 88 theme digestive 1669:1677 arg1 system 1679:1684 the human digestive system 1659:1684 the human digestive system 1659:1684 Taken together, our data imply that the NAALADL1 gene product's primary physiological function is associated with the final stages of protein/peptide digestion and absorption in the human digestive system. 25752612 4 89 theme first 778:782 arg1 evidence 797:804 the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 774:960 the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity 774:960 By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. 25752612 1 90 theme glutamate 213:221 arg1 carboxypeptidase 223:238 glutamate carboxypeptidase II 213:241 glutamate carboxypeptidase II 213:241 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 1 90 theme glutamate 213:221 arg1 metallopeptidase 246:261 a metallopeptidase 244:261 a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies 244:374 N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. 25752612 6 91 theme site-directed 1163:1175 arg1 mutagenesis 1177:1187 site-directed mutagenesis 1163:1187 site-directed mutagenesis 1163:1187 These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. 25752612 3 92 theme expression 699:708 arg1 profile 710:716 its expression profile 695:716 its expression profile 695:716 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 25752612 3 93 theme thorough 501:508 arg1 characterization 510:525 a thorough characterization 499:525 a thorough characterization 499:525 Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. 16732286 5 0 dep similar 817:823 arg1 including 866:874 including 866:874 including the location of the ligand-binding site within the Ig fold 866:933 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 16732286 3 1 theme factor 472:477 arg1 domains 479:485 the three epidermal growth factor domains 445:485 the three epidermal growth factor domains 445:485 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 5 2 theme site 911:914 arg1 location 880:887 the location 876:887 the location of the ligand-binding site within the Ig fold 876:933 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 16732286 4 3 from change 765:770 arg1 molecule 782:789 either molecule 775:789 either molecule 775:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 2 4 theme region 293:298 arg1 structures 255:264 the crystal structures 243:264 the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2 243:329 Here we present the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2. 16732286 2 5 with complex 313:319 arg1 Ang2 326:329 Ang2 326:329 Ang2 326:329 Here we present the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2. 16732286 4 6 theme little 743:748 arg1 change 765:770 little conformational change 743:770 little conformational change in either molecule 743:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 2 7 theme ligand-binding 278:291 arg1 region 293:298 the Tie2 ligand-binding region 269:298 the Tie2 ligand-binding region alone and in complex with Ang2 269:329 Here we present the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2. 16732286 3 8 dep compact 494:500 arg1 arrowhead-shaped 503:518 arrowhead-shaped 503:518 arrowhead-shaped 503:518 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 5 9 theme antigen 845:851 arg1 recognition 853:863 antibody-protein antigen recognition 828:863 antibody-protein antigen recognition 828:863 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 16732286 2 10 theme Tie2 273:276 arg1 region 293:298 the Tie2 ligand-binding region 269:298 the Tie2 ligand-binding region alone and in complex with Ang2 269:329 Here we present the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2. 16732286 5 11 theme Ang2-Tie2 792:800 arg1 recognition 802:812 Ang2-Tie2 recognition 792:812 Ang2-Tie2 recognition 792:812 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 16732286 3 12 theme immunoglobulin 391:404 arg1 domains 411:417 not two but three immunoglobulin (Ig) domains 373:417 domains 411:417 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 3 13 contain contains 364:371 arg2 Tie2 359:362 Tie2 359:362 Tie2 359:362 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 3 13 contain contains 364:371 arg1 Tie2 359:362 Tie2 359:362 Tie2 359:362 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 3 13 contain contains 364:371 arg2 three 385:389 three 385:389 three 385:389 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 3 13 contain contains 364:371 arg1 contrast 335:342 contrast 335:342 contrast to prediction 335:356 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 3 13 contain contains 364:371 arg2 domains 411:417 not two but three immunoglobulin (Ig) domains 373:417 domains 411:417 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 6 14 theme activation 1042:1051 arg1 mechanism 1020:1028 the mechanism 1016:1028 the mechanism of receptor activation 1016:1051 Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner. 16732286 1 15 theme central 164:170 arg1 roles 172:176 central roles 164:176 central roles 164:176 The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. 16732286 4 16 from rearrangements 724:737 arg1 molecule 782:789 either molecule 775:789 either molecule 775:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 4 17 theme domain 717:722 arg1 rearrangements 724:737 no domain rearrangements 714:737 no domain rearrangements 714:737 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 0 18 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of the Tie2 receptor ectodomain 0:49 Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex. 16732286 1 19 theme Tie 92:94 arg1 receptor 96:103 The Tie receptor 88:103 The Tie receptor tyrosine kinases 88:120 The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. 16732286 4 20 theme conformational 750:763 arg1 change 765:770 little conformational change 743:770 little conformational change in either molecule 743:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 4 21 theme receptor 635:642 arg1 kinase-where 644:655 a receptor kinase-where 633:655 a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule 633:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 5 22 theme antibody-protein 828:843 arg1 recognition 853:863 antibody-protein antigen recognition 828:863 antibody-protein antigen recognition 828:863 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 16732286 6 23 theme structure-based 967:981 arg1 mutagenesis 983:993 structure-based mutagenesis 967:993 structure-based mutagenesis 967:993 Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner. 16732286 3 24 theme compact 494:500 arg1 structure 520:528 a compact, arrowhead-shaped structure 492:528 a compact, arrowhead-shaped structure 492:528 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 1 25 theme tyrosine 105:112 arg1 kinases 114:120 tyrosine kinases 105:120 The Tie receptor tyrosine kinases 88:120 The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. 16732286 4 26 theme complementary 661:673 arg1 surfaces 675:682 two complementary surfaces 657:682 two complementary surfaces 657:682 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 0 27 theme receptor 31:38 arg1 ectodomain 40:49 the Tie2 receptor ectodomain 22:49 the Tie2 receptor ectodomain 22:49 Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex. 16732286 5 28 theme Ig 927:928 arg1 fold 930:933 the Ig fold 923:933 the Ig fold 923:933 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 16732286 4 29 theme recognition-unique 610:627 arg1 mode 595:598 a lock-and-key mode 580:598 a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule 580:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 1 30 theme tumor-induced 199:211 arg1 angiogenesis 213:224 tumor-induced angiogenesis 199:224 tumor-induced angiogenesis 199:224 The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. 16732286 0 31 theme Tie2 26:29 arg1 ectodomain 40:49 the Tie2 receptor ectodomain 22:49 the Tie2 receptor ectodomain 22:49 Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex. 16732286 6 32 theme structures 952:961 arg1 Analysis 936:943 Analysis 936:943 Analysis of the structures 936:961 Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner. 16732286 6 32 theme structures 952:961 arg1 mutagenesis 983:993 structure-based mutagenesis 967:993 structure-based mutagenesis 967:993 Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner. 16732286 6 33 theme receptor 1033:1040 arg1 activation 1042:1051 receptor activation 1033:1051 receptor activation 1033:1051 Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner. 16732286 4 34 theme ligand 603:608 arg1 recognition-unique 610:627 ligand recognition-unique 603:627 ligand recognition-unique 603:627 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 3 35 theme epidermal 455:463 arg1 factor 472:477 epidermal growth factor 455:477 the three epidermal growth factor domains 445:485 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 0 36 theme ectodomain 40:49 arg1 complex 79:85 the angiopoietin-2-Tie2 complex 55:85 the angiopoietin-2-Tie2 complex 55:85 Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex. 16732286 0 36 theme ectodomain 40:49 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of the Tie2 receptor ectodomain 0:49 Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex. 16732286 3 37 theme growth 465:470 arg1 factor 472:477 epidermal growth factor 455:477 the three epidermal growth factor domains 445:485 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 4 38 dep kinase-where 644:655 arg1 interact 684:691 interact 684:691 interact with each other with no domain rearrangements and little conformational change in either molecule 684:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 3 39 dep fold 426:429 arg1 domains 411:417 not two but three immunoglobulin (Ig) domains 373:417 domains 411:417 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 3 39 dep fold 426:429 arg1 three 385:389 three 385:389 three 385:389 In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. 16732286 6 40 theme similar 1140:1146 arg1 manner 1148:1153 a structurally similar manner 1125:1153 a structurally similar manner 1125:1153 Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner. 16732286 4 41 theme arrowhead 560:568 arg1 tip 549:551 the tip 545:551 the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule 545:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 0 42 theme angiopoietin-2-Tie2 59:77 arg1 complex 79:85 the angiopoietin-2-Tie2 complex 55:85 the angiopoietin-2-Tie2 complex 55:85 Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex. 16732286 2 43 theme crystal 247:253 arg1 structures 255:264 the crystal structures 243:264 the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2 243:329 Here we present the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2. 16732286 1 44 dep receptor 96:103 arg1 kinases 114:120 tyrosine kinases 105:120 The Tie receptor tyrosine kinases 88:120 The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. 16732286 1 44 dep receptor 96:103 arg1 ligands 151:157 ligands 151:157 ligands 151:157 The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. 16732286 4 45 theme lock-and-key 582:593 arg1 mode 595:598 a lock-and-key mode 580:598 a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule 580:789 Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition-unique for a receptor kinase-where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. 16732286 5 46 theme ligand-binding 896:909 arg1 site 911:914 the ligand-binding site 892:914 the ligand-binding site 892:914 Ang2-Tie2 recognition is similar to antibody-protein antigen recognition, including the location of the ligand-binding site within the Ig fold. 18585350 7 0 theme essential 1111:1119 arg1 component 1121:1129 an essential component 1108:1129 an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD 1108:1248 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 7 0 theme essential 1111:1119 arg1 CALHM1 1093:1098 CALHM1 1093:1098 CALHM1 1093:1098 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 0 1 from polymorphism 2:13 arg1 CALHM1 18:23 CALHM1 18:23 CALHM1 18:23 A polymorphism in CALHM1 influences Ca2+ homeostasis, Abeta levels, and Alzheimer's disease risk. 18585350 5 2 from AD 847:848 arg1 studies 878:884 independent case-control studies 853:884 independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)) 853:950 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 3 3 theme multipass 460:468 arg1 CALHM1 443:448 CALHM1 443:448 CALHM1 443:448 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 3 3 theme multipass 460:468 arg1 glycoprotein 484:495 a multipass transmembrane glycoprotein 458:495 a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels 458:558 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 3 4 theme cytosolic 511:519 arg1 2+ 524:525 2+ 524:525 2+ 524:525 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 3 4 theme cytosolic 511:519 arg1 Ca 521:522 cytosolic Ca 511:522 cytosolic Ca(2+) concentrations 511:541 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 6 5 theme Ca 1056:1057 arg1 permeability 1063:1074 CALHM1-mediated Ca(2+) permeability 1040:1074 CALHM1-mediated Ca(2+) permeability 1040:1074 We further found that the P86L polymorphism increases Abeta levels by interfering with CALHM1-mediated Ca(2+) permeability. 18585350 7 6 theme uncharacterized 1147:1161 arg1 channel 1179:1185 a previously uncharacterized cerebral Ca(2+) channel 1134:1185 a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD 1134:1248 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 3 7 theme Ca 521:522 arg1 concentrations 528:541 cytosolic Ca(2+) concentrations 511:541 cytosolic Ca(2+) concentrations 511:541 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 1 8 theme peptide 240:246 arg1 deposition 248:257 cerebral amyloid-beta (Abeta) peptide deposition 210:257 cerebral amyloid-beta (Abeta) peptide deposition 210:257 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 18585350 6 9 theme CALHM1-mediated 1040:1054 arg1 2+ 1059:1060 2+ 1059:1060 2+ 1059:1060 We further found that the P86L polymorphism increases Abeta levels by interfering with CALHM1-mediated Ca(2+) permeability. 18585350 6 9 theme CALHM1-mediated 1040:1054 arg1 Ca 1056:1057 CALHM1-mediated Ca 1040:1057 CALHM1-mediated Ca(2+) permeability 1040:1074 We further found that the P86L polymorphism increases Abeta levels by interfering with CALHM1-mediated Ca(2+) permeability. 18585350 5 10 theme P86L 784:787 arg1 polymorphism 789:800 the CALHM1 P86L polymorphism 773:800 the CALHM1 P86L polymorphism (rs2986017) 773:812 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 5 10 theme P86L 784:787 arg1 rs2986017 803:811 rs2986017 803:811 rs2986017 803:811 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 4 11 theme plasma 724:729 arg1 membrane 731:738 the plasma membrane 720:738 the plasma membrane 720:738 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 1 12 theme heterogeneous 140:152 arg1 disorder 154:161 a genetically heterogeneous disorder 126:161 a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition 126:257 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 18585350 1 12 theme heterogeneous 140:152 arg1 disease 110:116 Alzheimer's disease 98:116 Alzheimer's disease (AD) 98:121 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 18585350 4 13 theme selectivity 631:641 arg1 filter 643:648 the selectivity filter 627:648 the selectivity filter of the NMDA receptor 627:669 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 0 14 theme disease 84:90 arg1 risk 92:95 Alzheimer's disease risk 72:95 Alzheimer's disease risk 72:95 A polymorphism in CALHM1 influences Ca2+ homeostasis, Abeta levels, and Alzheimer's disease risk. 18585350 7 15 theme late-onset 1236:1245 arg1 AD 1247:1248 late-onset AD 1236:1248 late-onset AD 1236:1248 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 6 16 theme P86L 979:982 arg1 polymorphism 984:995 the P86L polymorphism 975:995 the P86L polymorphism 975:995 We further found that the P86L polymorphism increases Abeta levels by interfering with CALHM1-mediated Ca(2+) permeability. 18585350 2 17 theme linkage 359:365 arg1 regions 367:373 AD linkage regions 356:373 AD linkage regions 356:373 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 4 18 theme Ca 694:695 arg1 conductance 701:711 a large Ca(2+) conductance 686:711 a large Ca(2+) conductance across the plasma membrane 686:738 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 2 19 theme AD 356:357 arg1 regions 367:373 AD linkage regions 356:373 AD linkage regions 356:373 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 2 20 from located 345:351 arg1 regions 367:373 AD linkage regions 356:373 AD linkage regions 356:373 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 5 21 dep participants 894:905 arg1 = 937:937 = 937:937 = 937:937 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 4 22 theme sequence 600:607 arg1 similarities 609:620 strong sequence similarities 593:620 strong sequence similarities with the selectivity filter of the NMDA receptor 593:669 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 5 23 theme participants 894:905 arg1 studies 878:884 independent case-control studies 853:884 independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)) 853:950 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 4 24 theme receptor 662:669 arg1 filter 643:648 the selectivity filter 627:648 the selectivity filter of the NMDA receptor 627:669 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 4 25 theme strong 593:598 arg1 similarities 609:620 strong sequence similarities 593:620 strong sequence similarities with the selectivity filter of the NMDA receptor 593:669 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 0 26 theme Ca2+ 36:39 arg1 homeostasis 41:51 Ca2+ homeostasis 36:51 Ca2+ homeostasis 36:51 A polymorphism in CALHM1 influences Ca2+ homeostasis, Abeta levels, and Alzheimer's disease risk. 18585350 7 27 theme channel 1179:1185 arg1 component 1121:1129 an essential component 1108:1129 an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD 1108:1248 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 7 27 theme channel 1179:1185 arg1 CALHM1 1093:1098 CALHM1 1093:1098 CALHM1 1093:1098 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 7 28 theme cerebral 1163:1170 arg1 2+ 1175:1176 2+ 1175:1176 2+ 1175:1176 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 7 28 theme cerebral 1163:1170 arg1 Ca 1172:1173 cerebral Ca 1163:1173 a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD 1134:1248 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 1 29 theme early 180:184 arg1 atrophy 198:204 early hippocampal atrophy 180:204 early hippocampal atrophy 180:204 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 18585350 4 30 theme NMDA 657:660 arg1 receptor 662:669 the NMDA receptor 653:669 the NMDA receptor 653:669 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 1 31 theme hippocampal 186:196 arg1 atrophy 198:204 early hippocampal atrophy 180:204 early hippocampal atrophy 180:204 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 18585350 5 32 theme independent 853:863 arg1 studies 878:884 independent case-control studies 853:884 independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)) 853:950 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 3 33 theme Abeta 547:551 arg1 levels 553:558 Abeta levels 547:558 Abeta levels 547:558 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 5 34 theme case-control 865:876 arg1 studies 878:884 independent case-control studies 853:884 independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)) 853:950 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 4 35 with similarities 609:620 arg1 filter 643:648 the selectivity filter 627:648 the selectivity filter of the NMDA receptor 627:669 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 2 36 from regions 367:373 arg1 located 345:351 located 345:351 located 345:351 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 2 37 dep genes 291:295 arg1 located 345:351 located 345:351 located 345:351 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 2 37 dep genes 291:295 arg1 expressed 312:320 expressed 312:320 preferentially expressed in the hippocampus 297:339 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 3 38 gly glycoprotein 484:495 arg1 CALHM1 443:448 CALHM1 443:448 CALHM1 443:448 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 3 38 gly glycoprotein 484:495 arg1 glycoprotein 484:495 a multipass transmembrane glycoprotein 458:495 a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels 458:558 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 7 39 theme Ca 1172:1173 arg1 channel 1179:1185 a previously uncharacterized cerebral Ca(2+) channel 1134:1185 a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD 1134:1248 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 4 40 theme large 688:692 arg1 2+ 697:698 2+ 697:698 2+ 697:698 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 4 40 theme large 688:692 arg1 Ca 694:695 large Ca 688:695 a large Ca(2+) conductance across the plasma membrane 686:738 CALHM1 homomultimerizes, shares strong sequence similarities with the selectivity filter of the NMDA receptor, and generates a large Ca(2+) conductance across the plasma membrane. 18585350 5 41 dep = 937:937 arg1 1.44 929:932 1.44 929:932 1.44 929:932 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 5 42 theme CALHM1 777:782 arg1 polymorphism 789:800 the CALHM1 P86L polymorphism 773:800 the CALHM1 P86L polymorphism (rs2986017) 773:812 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 5 42 theme CALHM1 777:782 arg1 rs2986017 803:811 rs2986017 803:811 rs2986017 803:811 Importantly, we determined that the CALHM1 P86L polymorphism (rs2986017) is significantly associated with AD in independent case-control studies of 3404 participants (allele-specific OR = 1.44, p = 2 x 10(-10)). 18585350 3 43 theme transmembrane 470:482 arg1 CALHM1 443:448 CALHM1 443:448 CALHM1 443:448 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 3 43 theme transmembrane 470:482 arg1 glycoprotein 484:495 a multipass transmembrane glycoprotein 458:495 a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels 458:558 We show that CALHM1 encodes a multipass transmembrane glycoprotein that controls cytosolic Ca(2+) concentrations and Abeta levels. 18585350 7 44 theme Abeta 1201:1205 arg1 levels 1207:1212 Abeta levels 1201:1212 Abeta levels 1201:1212 We propose that CALHM1 encodes an essential component of a previously uncharacterized cerebral Ca(2+) channel that controls Abeta levels and susceptibility to late-onset AD. 18585350 1 45 theme cerebral 210:217 arg1 deposition 248:257 cerebral amyloid-beta (Abeta) peptide deposition 210:257 cerebral amyloid-beta (Abeta) peptide deposition 210:257 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 18585350 0 46 theme Abeta 54:58 arg1 levels 60:65 Abeta levels 54:65 Abeta levels 54:65 A polymorphism in CALHM1 influences Ca2+ homeostasis, Abeta levels, and Alzheimer's disease risk. 18585350 2 47 from gene 392:395 arg1 10q24.33 400:407 10q24.33 400:407 10q24.33 400:407 Using TissueInfo to screen for genes preferentially expressed in the hippocampus and located in AD linkage regions, we identified a gene on 10q24.33 that we call CALHM1. 18585350 6 48 theme Abeta 1007:1011 arg1 levels 1013:1018 Abeta levels 1007:1018 Abeta levels 1007:1018 We further found that the P86L polymorphism increases Abeta levels by interfering with CALHM1-mediated Ca(2+) permeability. 18585350 1 49 theme amyloid-beta 219:230 arg1 deposition 248:257 cerebral amyloid-beta (Abeta) peptide deposition 210:257 cerebral amyloid-beta (Abeta) peptide deposition 210:257 Alzheimer's disease (AD) is a genetically heterogeneous disorder characterized by early hippocampal atrophy and cerebral amyloid-beta (Abeta) peptide deposition. 9501084 5 0 theme collagen-binding 747:762 arg1 site 764:767 the collagen-binding site 743:767 the collagen-binding site 743:767 This constitutively activated mutant was used to map the collagen-binding site following alanine mutagenesis at 13 positions. 9501084 7 1 theme flat 1012:1015 arg1 ring 1017:1020 a flat ring 1010:1020 a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain 1010:1100 These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. 9501084 9 2 from consistent 1320:1329 arg1 form 1391:1394 a more cryptic form 1376:1394 a more cryptic form in intact BM-40 1376:1410 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 3 3 theme similar 495:501 arg1 affinity 486:493 collagen affinity 477:493 collagen affinity similar to that seen after proteolytic cleavage of this helix 477:555 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 1 4 theme moderate 260:267 arg1 affinity 269:276 moderate affinity 260:276 moderate affinity for several collagen types 260:303 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 0 5 theme epitope 83:89 arg1 mutagenesis 47:57 site-directed mutagenesis 33:57 site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin 33:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 5 6 used used 731:734 arg2 mutant 720:725 This constitutively activated mutant 690:725 This constitutively activated mutant 690:725 This constitutively activated mutant was used to map the collagen-binding site following alanine mutagenesis at 13 positions. 9501084 4 7 theme A 676:676 arg1 resolution 678:687 2.8 A resolution 672:687 2.8 A resolution 672:687 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 9501084 9 8 theme intact 1399:1404 arg1 BM-40 1406:1410 intact BM-40 1399:1410 intact BM-40 1399:1410 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 1 9 theme affinity 269:276 arg1 epitope 249:255 a binding epitope 239:255 a binding epitope of moderate affinity for several collagen types 239:303 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 0 10 theme activated 97:105 arg1 form 107:110 an activated form 94:110 an activated form of BM-40/SPARC/osteonectin 94:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 7 11 theme diameter 1030:1037 arg1 ring 1017:1020 a flat ring 1010:1020 a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain 1010:1100 These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. 9501084 9 12 theme same 1340:1343 arg1 location 1345:1352 the same location 1336:1352 the same location of the epitope 1336:1367 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 6 13 from N156 865:868 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 9 14 theme mutant 1275:1280 arg1 DeltaI 1282:1287 the non-activated mutant DeltaI 1257:1287 the non-activated mutant DeltaI 1257:1287 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 1 15 theme several 282:288 arg1 types 299:303 several collagen types 282:303 several collagen types 282:303 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 6 16 theme helix 873:877 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 3 17 theme 10-fold 457:463 arg1 increase 465:472 a 10-fold increase 455:472 a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix 455:555 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 4 18 theme mutant 662:667 arg1 structure 645:653 the crystal structure 633:653 the crystal structure of the mutant at 2.8 A resolution 633:687 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 9501084 9 19 theme epitope 1361:1367 arg1 location 1345:1352 the same location 1336:1352 the same location of the epitope 1336:1367 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 0 20 theme form 107:110 arg1 epitope 83:89 the collagen-binding epitope 62:89 the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin 62:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 7 21 theme A 1028:1028 arg1 diameter 1030:1037 15 A diameter 1025:1037 15 A diameter 1025:1037 These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. 9501084 3 22 from increase 465:472 arg1 affinity 486:493 collagen affinity 477:493 collagen affinity similar to that seen after proteolytic cleavage of this helix 477:555 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 3 23 theme proteolytic 522:532 arg1 cleavage 534:541 proteolytic cleavage 522:541 proteolytic cleavage of this helix 522:555 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 2 24 theme helix 389:393 arg1 alphaC 395:400 helix alphaC 389:400 helix alphaC 389:400 This epitope was predicted to reside in helix alphaA and to be partially masked by helix alphaC. 9501084 9 25 from mutations 1244:1252 arg1 DeltaI 1282:1287 the non-activated mutant DeltaI 1257:1287 the non-activated mutant DeltaI 1257:1287 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 6 26 from L242 891:894 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 6 27 from crucial 835:841 arg1 region 921:926 a loop region 914:926 a loop region connecting the two EF hands of BM-40 914:963 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 9 28 theme Selected 1235:1242 arg1 mutations 1244:1252 Selected mutations 1235:1252 Selected mutations in the non-activated mutant DeltaI 1235:1287 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 4 29 theme crystal 637:643 arg1 structure 645:653 the crystal structure 633:653 the crystal structure of the mutant at 2.8 A resolution 633:687 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 9501084 0 30 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure 0:16 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 1 31 theme collagen 290:297 arg1 types 299:303 several collagen types 282:303 several collagen types 282:303 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 32 dep positions 182:190 arg1 138-286 192:198 138-286 192:198 138-286 192:198 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 7 33 theme collagen 1086:1093 arg1 domain 1095:1100 a triple-helical collagen domain 1069:1100 a triple-helical collagen domain 1069:1100 These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. 9501084 9 34 theme non-activated 1261:1273 arg1 DeltaI 1282:1287 the non-activated mutant DeltaI 1257:1287 the non-activated mutant DeltaI 1257:1287 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 8 35 from sites 1210:1214 arg1 collagens 1224:1232 both collagens 1219:1232 both collagens 1219:1232 The mutations showed similar effects on binding to collagens I and IV, indicating nearly identical binding sites on both collagens. 9501084 1 36 theme matrix 208:213 arg1 BM-40 223:227 the matrix protein BM-40 204:227 the matrix protein BM-40 204:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 6 37 from R149 856:859 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 6 38 theme loop 916:919 arg1 region 921:926 a loop region 914:926 a loop region connecting the two EF hands of BM-40 914:963 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 0 39 theme BM-40/SPARC/osteonectin 115:137 arg1 form 107:110 an activated form 94:110 an activated form of BM-40/SPARC/osteonectin 94:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 3 40 theme helix 551:555 arg1 cleavage 534:541 proteolytic cleavage 522:541 proteolytic cleavage of this helix 522:555 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 7 41 theme triple-helical 1071:1084 arg1 domain 1095:1100 a triple-helical collagen domain 1069:1100 a triple-helical collagen domain 1069:1100 These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. 9501084 1 42 theme protein 215:221 arg1 BM-40 223:227 the matrix protein BM-40 204:227 the matrix protein BM-40 204:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 0 43 theme site-directed 33:45 arg1 mutagenesis 47:57 site-directed mutagenesis 33:57 site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin 33:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 3 44 theme alphaC 439:444 arg1 deletion 421:428 deletion 421:428 deletion of helix alphaC 421:444 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 5 45 theme activated 710:718 arg1 mutant 720:725 This constitutively activated mutant 690:725 This constitutively activated mutant 690:725 This constitutively activated mutant was used to map the collagen-binding site following alanine mutagenesis at 13 positions. 9501084 8 46 theme similar 1124:1130 arg1 effects 1132:1138 similar effects 1124:1138 similar effects 1124:1138 The mutations showed similar effects on binding to collagens I and IV, indicating nearly identical binding sites on both collagens. 9501084 1 47 theme BM-40 223:227 arg1 domain 174:179 The extracellular calcium-binding domain 140:179 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 140:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 47 theme BM-40 223:227 arg1 BM-40 223:227 the matrix protein BM-40 204:227 the matrix protein BM-40 204:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 47 theme BM-40 223:227 arg1 positions 182:190 positions 138-286 182:198 positions 138-286 182:198 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 5 48 theme alanine 779:785 arg1 mutagenesis 787:797 alanine mutagenesis 779:797 alanine mutagenesis 779:797 This constitutively activated mutant was used to map the collagen-binding site following alanine mutagenesis at 13 positions. 9501084 4 49 theme predicted 562:570 arg1 removal 572:578 The predicted removal 558:578 The predicted removal of the steric constraint 558:603 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 9501084 6 50 theme EF 947:948 arg1 hands 950:954 the two EF hands 939:954 the two EF hands of BM-40 939:963 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 6 51 from region 921:926 arg1 crucial 835:841 crucial 835:841 crucial 835:841 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 8 52 theme identical 1192:1200 arg1 sites 1210:1214 nearly identical binding sites 1185:1214 nearly identical binding sites on both collagens 1185:1232 The mutations showed similar effects on binding to collagens I and IV, indicating nearly identical binding sites on both collagens. 9501084 3 53 theme collagen 477:484 arg1 affinity 486:493 collagen affinity 477:493 collagen affinity similar to that seen after proteolytic cleavage of this helix 477:555 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 4 54 theme steric 587:592 arg1 constraint 594:603 the steric constraint 583:603 the steric constraint 583:603 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 9501084 9 55 theme collagen 1302:1309 arg1 binding 1311:1317 collagen binding 1302:1317 collagen binding 1302:1317 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 6 56 from M245 897:900 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 1 57 contain possesses 229:237 arg1 domain 174:179 The extracellular calcium-binding domain 140:179 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 140:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 57 contain possesses 229:237 arg1 positions 182:190 positions 138-286 182:198 positions 138-286 182:198 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 57 contain possesses 229:237 arg1 BM-40 223:227 the matrix protein BM-40 204:227 the matrix protein BM-40 204:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 57 contain possesses 229:237 arg2 epitope 249:255 a binding epitope 239:255 a binding epitope of moderate affinity for several collagen types 239:303 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 9 58 theme cryptic 1383:1389 arg1 form 1391:1394 a more cryptic form 1376:1394 a more cryptic form in intact BM-40 1376:1410 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 2 59 theme helix 346:350 arg1 alphaA 352:357 helix alphaA 346:357 helix alphaA 346:357 This epitope was predicted to reside in helix alphaA and to be partially masked by helix alphaC. 9501084 8 60 theme binding 1202:1208 arg1 sites 1210:1214 nearly identical binding sites 1185:1214 nearly identical binding sites on both collagens 1185:1232 The mutations showed similar effects on binding to collagens I and IV, indicating nearly identical binding sites on both collagens. 9501084 6 61 from binding 847:853 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 9 62 with consistent 1320:1329 arg1 location 1345:1352 the same location 1336:1352 the same location of the epitope 1336:1367 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 4 63 theme constraint 594:603 arg1 removal 572:578 The predicted removal 558:578 The predicted removal of the steric constraint 558:603 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 9501084 6 64 from E246 906:909 arg1 alphaA 879:884 helix alphaA 873:884 helix alphaA 873:884 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 9 65 from form 1391:1394 arg1 BM-40 1406:1410 intact BM-40 1399:1410 intact BM-40 1399:1410 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 9 65 from form 1391:1394 arg1 consistent 1320:1329 consistent 1320:1329 consistent 1320:1329 Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40. 9501084 1 66 theme extracellular 144:156 arg1 domain 174:179 The extracellular calcium-binding domain 140:179 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 140:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 66 theme extracellular 144:156 arg1 BM-40 223:227 the matrix protein BM-40 204:227 the matrix protein BM-40 204:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 66 theme extracellular 144:156 arg1 positions 182:190 positions 138-286 182:198 positions 138-286 182:198 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 67 theme binding 241:247 arg1 epitope 249:255 a binding epitope 239:255 a binding epitope of moderate affinity for several collagen types 239:303 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 0 68 theme collagen-binding 66:81 arg1 epitope 83:89 the collagen-binding epitope 62:89 the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin 62:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 0 69 dep structure 8:16 arg1 mutagenesis 47:57 site-directed mutagenesis 33:57 site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin 33:137 Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin. 9501084 1 70 theme calcium-binding 158:172 arg1 domain 174:179 The extracellular calcium-binding domain 140:179 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 140:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 70 theme calcium-binding 158:172 arg1 BM-40 223:227 the matrix protein BM-40 204:227 the matrix protein BM-40 204:227 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 1 70 theme calcium-binding 158:172 arg1 positions 182:190 positions 138-286 182:198 positions 138-286 182:198 The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. 9501084 3 71 theme helix 433:437 arg1 alphaC 439:444 helix alphaC 433:444 helix alphaC 433:444 Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. 9501084 6 72 theme BM-40 959:963 arg1 hands 950:954 the two EF hands 939:954 the two EF hands of BM-40 939:963 Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. 9501084 7 73 theme domain 1095:1100 arg1 diameter 1057:1064 the diameter 1053:1064 the diameter of a triple-helical collagen domain 1053:1100 These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. 9501084 4 74 from resolution 678:687 arg1 structure 645:653 the crystal structure 633:653 the crystal structure of the mutant at 2.8 A resolution 633:687 The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. 31471319 5 0 theme zinc 727:730 arg1 function 739:746 its zinc efflux function 723:746 its zinc efflux function 723:746 We observed that under zinc-sufficient conditions, ZNT1 accumulates on the plasma membrane, consistent with its zinc efflux function. 31471319 6 1 theme proteasomal 880:890 arg1 pathways 906:913 both the proteasomal and lysosomal pathways 871:913 pathways 906:913 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 10 2 theme homeostasis 1623:1633 arg1 sophisticated 1685:1697 sophisticated 1685:1697 sophisticated 1685:1697 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 10 2 theme homeostasis 1623:1633 arg1 control 1585:1591 the control 1581:1591 the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression 1581:1675 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 9 3 theme high 1414:1417 arg1 levels 1424:1429 high zinc levels 1414:1429 high zinc levels 1414:1429 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 31471319 5 4 theme efflux 732:737 arg1 function 739:746 its zinc efflux function 723:746 its zinc efflux function 723:746 We observed that under zinc-sufficient conditions, ZNT1 accumulates on the plasma membrane, consistent with its zinc efflux function. 31471319 4 5 from regulation 465:474 arg1 cells 519:523 human and vertebrate cells 498:523 human and vertebrate cells 498:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 5 6 theme plasma 690:695 arg1 membrane 697:704 the plasma membrane 686:704 the plasma membrane 686:704 We observed that under zinc-sufficient conditions, ZNT1 accumulates on the plasma membrane, consistent with its zinc efflux function. 31471319 6 7 theme zinc-deficient 768:781 arg1 conditions 783:792 zinc-deficient conditions 768:792 zinc-deficient conditions 768:792 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 4 8 theme ZNT1 541:544 arg1 expression 546:555 ZNT1 expression 541:555 ZNT1 expression 541:555 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 8 9 theme domains 1172:1178 arg1 V 1180:1180 transmembrane domains V and VI 1158:1187 V 1180:1180 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 9 10 theme posttranslational 1313:1329 arg1 modification 1331:1342 this posttranslational modification 1308:1342 this posttranslational modification 1308:1342 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 31471319 7 11 theme ZNT1 932:935 arg1 expression 937:946 Zinc-responsive ZNT1 expression 916:946 Zinc-responsive ZNT1 expression 916:946 Zinc-responsive ZNT1 expression corresponded with that of metallothionein, supporting the idea that ZNT1 and metallothionein cooperatively regulate cellular zinc homeostasis. 31471319 9 12 theme zinc 1419:1422 arg1 levels 1424:1429 high zinc levels 1414:1429 high zinc levels 1414:1429 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 31471319 0 13 theme cellular 86:93 arg1 levels 100:105 cellular zinc levels 86:105 cellular zinc levels 86:105 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 0 14 from expression 26:35 arg1 surface 49:55 the cell surface 40:55 the cell surface 40:55 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 8 15 theme stability 1248:1256 arg1 regulation 1229:1238 the regulation 1225:1238 the regulation of ZNT1 stability 1225:1256 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 8 16 theme ZNT1 1243:1246 arg1 stability 1248:1256 ZNT1 stability 1243:1256 ZNT1 stability 1243:1256 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 2 17 theme pathological 347:358 arg1 functions 360:368 the physiological and pathological functions 325:368 the physiological and pathological functions of ZNT1 325:376 Numerous studies have focused on the physiological and pathological functions of ZNT1. 31471319 10 18 theme zinc 1618:1621 arg1 homeostasis 1623:1633 cellular and systemic zinc homeostasis 1596:1633 homeostasis 1623:1633 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 4 19 from cells 519:523 arg1 regulation 465:474 the regulation 461:474 the regulation of ZNT1 expression in human and vertebrate cells 461:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 3 20 theme biochemical 392:402 arg1 features 404:411 its biochemical features 388:411 its biochemical features 388:411 However, its biochemical features remain poorly understood. 31471319 0 21 theme transporter 5:15 arg1 expression 26:35 Zinc transporter 1 (ZNT1) expression 0:35 Zinc transporter 1 (ZNT1) expression on the cell surface 0:55 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 10 22 theme zinc 1545:1548 arg1 homeostasis 1550:1560 cellular zinc homeostasis 1536:1560 cellular zinc homeostasis 1536:1560 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 7 23 theme Zinc-responsive 916:930 arg1 expression 937:946 Zinc-responsive ZNT1 expression 916:946 Zinc-responsive ZNT1 expression 916:946 Zinc-responsive ZNT1 expression corresponded with that of metallothionein, supporting the idea that ZNT1 and metallothionein cooperatively regulate cellular zinc homeostasis. 31471319 8 24 gly N-glycosylated 1099:1112 arg1 Asn299 1117:1122 Asn299 1117:1122 Asn299 in the extracellular loop between transmembrane domains V and VI 1117:1187 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 8 24 gly N-glycosylated 1099:1112 arg2 Asn299 1117:1122 Asn299 1117:1122 Asn299 in the extracellular loop between transmembrane domains V and VI 1117:1187 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 8 24 gly N-glycosylated 1099:1112 arg1 ZNT1 1091:1094 ZNT1 1091:1094 ZNT1 1091:1094 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 8 24 gly N-glycosylated 1099:1112 arg2 ZNT1 1091:1094 ZNT1 1091:1094 ZNT1 1091:1094 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 2 25 theme physiological 329:341 arg1 functions 360:368 the physiological and pathological functions 325:368 the physiological and pathological functions of ZNT1 325:376 Numerous studies have focused on the physiological and pathological functions of ZNT1. 31471319 0 26 theme Zinc 0:3 arg1 transporter 5:15 Zinc transporter 1 0:17 Zinc transporter 1 (ZNT1) expression on the cell surface 0:55 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 0 26 theme Zinc 0:3 arg1 ZNT1 20:23 ZNT1 20:23 ZNT1 20:23 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 4 27 theme vertebrate 508:517 arg1 cells 519:523 human and vertebrate cells 498:523 human and vertebrate cells 498:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 8 28 gly nonglycosylated 1262:1276 arg1 ZNT1 1278:1281 nonglycosylated ZNT1 1262:1281 nonglycosylated ZNT1 1262:1281 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 8 29 theme transmembrane 1158:1170 arg1 domains 1172:1178 transmembrane domains 1158:1178 transmembrane domains 1158:1178 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 1 30 theme only 141:144 arg1 transporter 151:161 the only zinc transporter 137:161 the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space 137:289 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 1 30 theme only 141:144 arg1 transporter 113:123 Zinc transporter 1 108:125 Zinc transporter 1 (ZNT1) 108:132 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 10 31 theme cellular 1596:1603 arg1 homeostasis 1623:1633 cellular and systemic zinc homeostasis 1596:1633 homeostasis 1623:1633 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 1 32 theme pivotal 226:232 arg1 role 234:237 a pivotal role 224:237 a pivotal role 224:237 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 7 33 theme zinc 1073:1076 arg1 homeostasis 1078:1088 cellular zinc homeostasis 1064:1088 cellular zinc homeostasis 1064:1088 Zinc-responsive ZNT1 expression corresponded with that of metallothionein, supporting the idea that ZNT1 and metallothionein cooperatively regulate cellular zinc homeostasis. 31471319 1 34 theme zinc 146:149 arg1 transporter 151:161 the only zinc transporter 137:161 the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space 137:289 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 1 34 theme zinc 146:149 arg1 transporter 113:123 Zinc transporter 1 108:125 Zinc transporter 1 (ZNT1) 108:132 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 0 35 theme zinc 95:98 arg1 levels 100:105 cellular zinc levels 86:105 cellular zinc levels 86:105 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 6 36 dep pathways 906:913 arg1 both 871:874 both 871:874 both 871:874 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 2 37 theme ZNT1 373:376 arg1 functions 360:368 the physiological and pathological functions 325:368 the physiological and pathological functions of ZNT1 325:376 Numerous studies have focused on the physiological and pathological functions of ZNT1. 31471319 10 38 theme dynamic 1639:1645 arg1 regulation 1647:1656 dynamic regulation 1639:1656 dynamic regulation of ZNT1 expression 1639:1675 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 10 39 theme molecular 1484:1492 arg1 insights 1494:1501 molecular insights 1484:1501 molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis 1484:1560 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 9 40 theme subcellular 1438:1448 arg1 localization 1450:1461 its subcellular localization 1434:1461 its subcellular localization 1434:1461 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 31471319 8 41 theme nonglycosylated 1262:1276 arg1 ZNT1 1278:1281 nonglycosylated ZNT1 1262:1281 nonglycosylated ZNT1 1262:1281 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 10 42 theme expression 1666:1675 arg1 regulation 1647:1656 dynamic regulation 1639:1656 dynamic regulation of ZNT1 expression 1639:1675 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 1 43 theme cytosolic 249:257 arg1 zinc 259:262 cytosolic zinc 249:262 cytosolic zinc 249:262 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 7 44 theme cellular 1064:1071 arg1 homeostasis 1078:1088 cellular zinc homeostasis 1064:1088 cellular zinc homeostasis 1064:1088 Zinc-responsive ZNT1 expression corresponded with that of metallothionein, supporting the idea that ZNT1 and metallothionein cooperatively regulate cellular zinc homeostasis. 31471319 6 45 theme plasma 817:822 arg1 membrane 824:831 the plasma membrane 813:831 the plasma membrane 813:831 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 5 46 with consistent 707:716 arg1 function 739:746 its zinc efflux function 723:746 its zinc efflux function 723:746 We observed that under zinc-sufficient conditions, ZNT1 accumulates on the plasma membrane, consistent with its zinc efflux function. 31471319 4 47 theme zinc 602:605 arg1 status 607:612 cellular zinc status 593:612 cellular zinc status 593:612 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 4 48 from expression 484:493 arg1 cells 519:523 human and vertebrate cells 498:523 human and vertebrate cells 498:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 4 49 theme ZNT1 479:482 arg1 expression 484:493 ZNT1 expression 479:493 ZNT1 expression in human and vertebrate cells 479:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 10 50 theme cellular 1536:1543 arg1 homeostasis 1550:1560 cellular zinc homeostasis 1536:1560 cellular zinc homeostasis 1536:1560 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 4 51 theme expression 484:493 arg1 regulation 465:474 the regulation 461:474 the regulation of ZNT1 expression in human and vertebrate cells 461:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 9 52 contain had 1344:1346 arg1 modification 1331:1342 this posttranslational modification 1308:1342 this posttranslational modification 1308:1342 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 31471319 9 52 contain had 1344:1346 arg2 effect 1351:1356 no effect 1348:1356 no effect 1348:1356 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 31471319 2 53 theme Numerous 292:299 arg1 studies 301:307 Numerous studies 292:307 Numerous studies 292:307 Numerous studies have focused on the physiological and pathological functions of ZNT1. 31471319 6 54 from molecules 800:808 arg1 membrane 824:831 the plasma membrane 813:831 the plasma membrane 813:831 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 4 55 theme human 498:502 arg1 cells 519:523 human and vertebrate cells 498:523 human and vertebrate cells 498:523 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 5 56 theme zinc-sufficient 638:652 arg1 conditions 654:663 zinc-sufficient conditions 638:663 zinc-sufficient conditions 638:663 We observed that under zinc-sufficient conditions, ZNT1 accumulates on the plasma membrane, consistent with its zinc efflux function. 31471319 10 57 theme homeostasis 1550:1560 arg1 regulation 1522:1531 ZNT1-mediated regulation 1508:1531 ZNT1-mediated regulation of cellular zinc homeostasis 1508:1560 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 6 58 theme lysosomal 896:904 arg1 pathways 906:913 both the proteasomal and lysosomal pathways 871:913 pathways 906:913 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 8 59 from Asn299 1117:1122 arg1 loop 1145:1148 the extracellular loop 1127:1148 the extracellular loop between transmembrane domains V and VI 1127:1187 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 10 60 theme ZNT1-mediated 1508:1520 arg1 regulation 1522:1531 ZNT1-mediated regulation 1508:1531 ZNT1-mediated regulation of cellular zinc homeostasis 1508:1560 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 6 61 theme ZNT1 795:798 arg1 molecules 800:808 ZNT1 molecules 795:808 ZNT1 molecules on the plasma membrane 795:831 In contrast, under zinc-deficient conditions, ZNT1 molecules on the plasma membrane were endocytosed and degraded through both the proteasomal and lysosomal pathways. 31471319 1 62 theme Zinc 108:111 arg1 transporter 151:161 the only zinc transporter 137:161 the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space 137:289 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 1 62 theme Zinc 108:111 arg1 transporter 113:123 Zinc transporter 1 108:125 Zinc transporter 1 (ZNT1) 108:132 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 1 62 theme Zinc 108:111 arg1 ZNT1 128:131 ZNT1 128:131 ZNT1 128:131 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 0 63 theme cell 44:47 arg1 surface 49:55 the cell surface 40:55 the cell surface 40:55 Zinc transporter 1 (ZNT1) expression on the cell surface is elaborately controlled by cellular zinc levels. 31471319 1 64 theme located 177:183 arg1 transporter 151:161 the only zinc transporter 137:161 the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space 137:289 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 1 64 theme located 177:183 arg1 transporter 113:123 Zinc transporter 1 108:125 Zinc transporter 1 (ZNT1) 108:132 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 4 65 theme cellular 593:600 arg1 status 607:612 cellular zinc status 593:612 cellular zinc status 593:612 Here, we investigated the regulation of ZNT1 expression in human and vertebrate cells, and found that ZNT1 expression is posttranslationally regulated by cellular zinc status. 31471319 10 66 theme ZNT1 1661:1664 arg1 expression 1666:1675 ZNT1 expression 1661:1675 ZNT1 expression 1661:1675 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 1 67 theme plasma 192:197 arg1 membrane 199:206 the plasma membrane 188:206 the plasma membrane 188:206 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 8 68 theme extracellular 1131:1143 arg1 loop 1145:1148 the extracellular loop 1127:1148 the extracellular loop between transmembrane domains V and VI 1127:1187 ZNT1 is N-glycosylated on Asn299 in the extracellular loop between transmembrane domains V and VI, and this appears to be involved in the regulation of ZNT1 stability, as nonglycosylated ZNT1 is more stable. 31471319 1 69 theme extracellular 271:283 arg1 space 285:289 the extracellular space 267:289 the extracellular space 267:289 Zinc transporter 1 (ZNT1) is the only zinc transporter predominantly located on the plasma membrane, where it plays a pivotal role exporting cytosolic zinc to the extracellular space. 31471319 10 70 theme systemic 1609:1616 arg1 homeostasis 1623:1633 cellular and systemic zinc homeostasis 1596:1633 homeostasis 1623:1633 Our results provide molecular insights into ZNT1-mediated regulation of cellular zinc homeostasis, and indicate that the control of cellular and systemic zinc homeostasis via dynamic regulation of ZNT1 expression is more sophisticated than previously thought. 31471319 9 71 theme cellular 1386:1393 arg1 resistance 1395:1404 cellular resistance 1386:1404 cellular resistance against high zinc levels or its subcellular localization 1386:1461 However, this posttranslational modification had no effect on ZNT1's ability to confer cellular resistance against high zinc levels or its subcellular localization. 9690478 0 0 theme growth 72:77 arg1 receptor 86:93 the type-1 insulin-like growth factor receptor 48:93 the type-1 insulin-like growth factor receptor 48:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 4 1 theme receptor 491:498 arg1 family 507:512 The epidermal growth-factor receptor (EGFR) family 463:512 The epidermal growth-factor receptor (EGFR) family 463:512 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 4 1 theme receptor 491:498 arg1 related 525:531 related 525:531 related 525:531 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 1 2 theme receptor 229:236 arg1 superfamily 238:248 the tyrosine-kinase receptor superfamily 209:248 the tyrosine-kinase receptor superfamily 209:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 0 3 theme insulin-like 59:70 arg1 receptor 86:93 the type-1 insulin-like growth factor receptor 48:93 the type-1 insulin-like growth factor receptor 48:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 2 4 theme normal 327:332 arg1 growth 334:339 normal growth 327:339 normal growth 327:339 IR is essential for glucose homeostasis, whereas IGF-1R is involved in both normal growth and development and malignant transformation. 9690478 1 5 theme superfamily 238:248 arg1 members 198:204 closely related members 182:204 closely related members of the tyrosine-kinase receptor superfamily 182:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 5 theme superfamily 238:248 arg1 receptor 134:141 The type-1 insulin-like growth-factor receptor 96:141 The type-1 insulin-like growth-factor receptor (IGF-1R) 96:150 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 5 theme superfamily 238:248 arg1 receptor 164:171 insulin receptor 156:171 insulin receptor (IR) 156:176 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 0 6 theme receptor 86:93 arg1 receptor 86:93 the type-1 insulin-like growth factor receptor 48:93 the type-1 insulin-like growth factor receptor 48:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 0 6 theme receptor 86:93 arg1 domains 37:43 the first three domains 21:43 the first three domains of the type-1 insulin-like growth factor receptor 21:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 8 7 theme central 1006:1012 arg1 space 1014:1018 a central space 1004:1018 a central space of sufficient size to accommodate a ligand molecule 1004:1070 The three domains surround a central space of sufficient size to accommodate a ligand molecule. 9690478 2 8 theme glucose 271:277 arg1 homeostasis 279:289 glucose homeostasis 271:289 glucose homeostasis 271:289 IR is essential for glucose homeostasis, whereas IGF-1R is involved in both normal growth and development and malignant transformation. 9690478 4 9 theme sequence 570:577 arg1 identity 579:586 significant sequence identity 558:586 significant sequence identity 558:586 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 0 10 theme factor 79:84 arg1 receptor 86:93 the type-1 insulin-like growth factor receptor 48:93 the type-1 insulin-like growth factor receptor 48:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 7 11 theme rod-shaped 913:922 arg1 domain 924:929 a rod-shaped domain 911:929 a rod-shaped domain 911:929 The Cys-rich region is composed of eight disulphide-bonded modules, seven of which form a rod-shaped domain with modules associated in an unusual manner. 9690478 7 12 theme Cys-rich 827:834 arg1 region 836:841 The Cys-rich region 823:841 The Cys-rich region 823:841 The Cys-rich region is composed of eight disulphide-bonded modules, seven of which form a rod-shaped domain with modules associated in an unusual manner. 9690478 4 13 theme significant 558:568 arg1 identity 579:586 significant sequence identity 558:586 significant sequence identity 558:586 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 5 14 theme A 736:736 arg1 resolution 738:747 2.6 A resolution 732:747 2.6 A resolution 732:747 We now present the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution. 9690478 9 15 dep residues 1096:1103 arg1 1-462 1105:1109 1-462 1105:1109 1-462 1105:1109 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 1 16 theme insulin 156:162 arg1 members 198:204 closely related members 182:204 closely related members of the tyrosine-kinase receptor superfamily 182:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 16 theme insulin 156:162 arg1 IR 174:175 IR 174:175 IR 174:175 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 16 theme insulin 156:162 arg1 receptor 164:171 insulin receptor 156:171 insulin receptor (IR) 156:176 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 16 theme insulin 156:162 arg1 receptor 134:141 The type-1 insulin-like growth-factor receptor 96:141 The type-1 insulin-like growth-factor receptor (IGF-1R) 96:150 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 4 17 theme growth-factor 477:489 arg1 EGFR 501:504 EGFR 501:504 EGFR 501:504 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 4 17 theme growth-factor 477:489 arg1 receptor 491:498 epidermal growth-factor receptor 467:498 The epidermal growth-factor receptor (EGFR) family 463:512 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 6 18 theme L 754:754 arg1 domains 756:762 The L domains 750:762 The L domains each 750:767 The L domains each consist of a single-stranded right-handed beta-helix. 9690478 4 19 theme epidermal 467:475 arg1 EGFR 501:504 EGFR 501:504 EGFR 501:504 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 4 19 theme epidermal 467:475 arg1 receptor 491:498 epidermal growth-factor receptor 467:498 The epidermal growth-factor receptor (EGFR) family 463:512 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 8 20 theme size 1034:1037 arg1 space 1014:1018 a central space 1004:1018 a central space of sufficient size to accommodate a ligand molecule 1004:1070 The three domains surround a central space of sufficient size to accommodate a ligand molecule. 9690478 9 21 theme determinants 1146:1157 arg1 many 1134:1137 many 1134:1137 many 1134:1137 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 9 21 theme determinants 1146:1157 arg1 determinants 1146:1157 the determinants 1142:1157 the determinants responsible for hormone binding and ligand specificity 1142:1212 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 2 22 theme malignant 361:369 arg1 transformation 371:384 malignant transformation 361:384 malignant transformation 361:384 IR is essential for glucose homeostasis, whereas IGF-1R is involved in both normal growth and development and malignant transformation. 9690478 0 23 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 0:94 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 8 24 theme sufficient 1023:1032 arg1 size 1034:1037 sufficient size 1023:1037 sufficient size 1023:1037 The three domains surround a central space of sufficient size to accommodate a ligand molecule. 9690478 3 25 located found 421:425 arg1 animals 430:436 animals 430:436 animals as simple as cnidarians 430:460 Homologues of these receptors are found in animals as simple as cnidarians. 9690478 3 25 located found 421:425 arg2 Homologues 387:396 Homologues 387:396 Homologues of these receptors 387:415 Homologues of these receptors are found in animals as simple as cnidarians. 9690478 9 26 theme responsible 1159:1169 arg1 determinants 1146:1157 the determinants 1142:1157 the determinants responsible for hormone binding and ligand specificity 1142:1212 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 5 27 theme first 671:675 arg1 IGF-IR 694:699 IGF-IR 694:699 IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution 694:747 We now present the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution. 9690478 5 27 theme first 671:675 arg1 domains 683:689 the first three domains 667:689 the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution 667:747 We now present the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution. 9690478 4 28 dep portion 609:615 arg1 describe 620:627 describe 620:627 describe here 620:632 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 6 29 dep domains 756:762 arg1 each 764:767 each 764:767 each 764:767 The L domains each consist of a single-stranded right-handed beta-helix. 9690478 3 30 theme as 448:449 arg1 cnidarians 451:460 simple as cnidarians 441:460 simple as cnidarians 441:460 Homologues of these receptors are found in animals as simple as cnidarians. 9690478 9 31 theme hormone 1175:1181 arg1 binding 1183:1189 hormone binding 1175:1189 hormone binding 1175:1189 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 4 32 theme IR 540:541 arg1 family 543:548 the IR family 536:548 the IR family 536:548 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 5 33 theme domains 683:689 arg1 structure 654:662 the structure 650:662 the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution 650:747 We now present the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution. 9690478 3 34 theme receptors 407:415 arg1 Homologues 387:396 Homologues 387:396 Homologues of these receptors 387:415 Homologues of these receptors are found in animals as simple as cnidarians. 9690478 0 35 theme first 25:29 arg1 receptor 86:93 the type-1 insulin-like growth factor receptor 48:93 the type-1 insulin-like growth factor receptor 48:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 0 35 theme first 25:29 arg1 domains 37:43 the first three domains 21:43 the first three domains of the type-1 insulin-like growth factor receptor 21:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 1 36 theme related 190:196 arg1 members 198:204 closely related members 182:204 closely related members of the tyrosine-kinase receptor superfamily 182:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 36 theme related 190:196 arg1 receptor 134:141 The type-1 insulin-like growth-factor receptor 96:141 The type-1 insulin-like growth-factor receptor (IGF-1R) 96:150 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 36 theme related 190:196 arg1 receptor 164:171 insulin receptor 156:171 insulin receptor (IR) 156:176 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 5 37 theme IGF-IR 694:699 arg1 IGF-IR 694:699 IGF-IR 694:699 IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution 694:747 We now present the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution. 9690478 5 37 theme IGF-IR 694:699 arg1 domains 683:689 the first three domains 667:689 the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution 667:747 We now present the structure of the first three domains of IGF-IR (L1-Cys-rich-L2) determined to 2.6 A resolution. 9690478 9 38 theme ligand 1195:1200 arg1 specificity 1202:1212 ligand specificity 1195:1212 ligand specificity 1195:1212 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 1 39 theme type-1 100:105 arg1 IGF-1R 144:149 IGF-1R 144:149 IGF-1R 144:149 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 39 theme type-1 100:105 arg1 members 198:204 closely related members 182:204 closely related members of the tyrosine-kinase receptor superfamily 182:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 39 theme type-1 100:105 arg1 receptor 134:141 The type-1 insulin-like growth-factor receptor 96:141 The type-1 insulin-like growth-factor receptor (IGF-1R) 96:150 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 39 theme type-1 100:105 arg1 receptor 164:171 insulin receptor 156:171 insulin receptor (IR) 156:176 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 0 40 theme domains 37:43 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 0:94 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 4 41 contain has 554:556 arg1 family 507:512 The epidermal growth-factor receptor (EGFR) family 463:512 The epidermal growth-factor receptor (EGFR) family 463:512 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 4 41 contain has 554:556 arg1 related 525:531 related 525:531 related 525:531 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 4 41 contain has 554:556 arg2 identity 579:586 significant sequence identity 558:586 significant sequence identity 558:586 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 10 42 theme IR 1279:1280 arg1 subfamily 1282:1290 the IR subfamily 1275:1290 the IR subfamily 1275:1290 This structure therefore shows how the IR subfamily might interact with their ligands. 9690478 1 43 theme insulin-like 107:118 arg1 IGF-1R 144:149 IGF-1R 144:149 IGF-1R 144:149 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 43 theme insulin-like 107:118 arg1 members 198:204 closely related members 182:204 closely related members of the tyrosine-kinase receptor superfamily 182:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 43 theme insulin-like 107:118 arg1 receptor 134:141 The type-1 insulin-like growth-factor receptor 96:141 The type-1 insulin-like growth-factor receptor (IGF-1R) 96:150 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 43 theme insulin-like 107:118 arg1 receptor 164:171 insulin receptor 156:171 insulin receptor (IR) 156:176 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 8 44 theme ligand 1056:1061 arg1 molecule 1063:1070 a ligand molecule 1054:1070 a ligand molecule 1054:1070 The three domains surround a central space of sufficient size to accommodate a ligand molecule. 9690478 6 45 theme right-handed 798:809 arg1 beta-helix 811:820 a single-stranded right-handed beta-helix 780:820 a single-stranded right-handed beta-helix 780:820 The L domains each consist of a single-stranded right-handed beta-helix. 9690478 1 46 theme growth-factor 120:132 arg1 IGF-1R 144:149 IGF-1R 144:149 IGF-1R 144:149 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 46 theme growth-factor 120:132 arg1 members 198:204 closely related members 182:204 closely related members of the tyrosine-kinase receptor superfamily 182:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 46 theme growth-factor 120:132 arg1 receptor 134:141 The type-1 insulin-like growth-factor receptor 96:141 The type-1 insulin-like growth-factor receptor (IGF-1R) 96:150 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 1 46 theme growth-factor 120:132 arg1 receptor 164:171 insulin receptor 156:171 insulin receptor (IR) 156:176 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 9690478 6 47 theme single-stranded 782:796 arg1 beta-helix 811:820 a single-stranded right-handed beta-helix 780:820 a single-stranded right-handed beta-helix 780:820 The L domains each consist of a single-stranded right-handed beta-helix. 9690478 7 48 theme disulphide-bonded 864:880 arg1 modules 882:888 eight disulphide-bonded modules 858:888 eight disulphide-bonded modules 858:888 The Cys-rich region is composed of eight disulphide-bonded modules, seven of which form a rod-shaped domain with modules associated in an unusual manner. 9690478 0 49 theme type-1 52:57 arg1 receptor 86:93 the type-1 insulin-like growth factor receptor 48:93 the type-1 insulin-like growth factor receptor 48:93 Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor. 9690478 3 50 theme simple 441:446 arg1 cnidarians 451:460 simple as cnidarians 441:460 simple as cnidarians 441:460 Homologues of these receptors are found in animals as simple as cnidarians. 9690478 7 51 theme unusual 961:967 arg1 manner 969:974 an unusual manner 958:974 an unusual manner 958:974 The Cys-rich region is composed of eight disulphide-bonded modules, seven of which form a rod-shaped domain with modules associated in an unusual manner. 9690478 2 52 dep essential 257:265 arg1 whereas 292:298 whereas 292:298 whereas 292:298 IR is essential for glucose homeostasis, whereas IGF-1R is involved in both normal growth and development and malignant transformation. 9690478 7 53 theme modules 882:888 arg1 modules 882:888 eight disulphide-bonded modules 858:888 eight disulphide-bonded modules 858:888 The Cys-rich region is composed of eight disulphide-bonded modules, seven of which form a rod-shaped domain with modules associated in an unusual manner. 9690478 7 53 theme modules 882:888 arg1 seven 891:895 seven 891:895 seven 891:895 The Cys-rich region is composed of eight disulphide-bonded modules, seven of which form a rod-shaped domain with modules associated in an unusual manner. 9690478 4 54 theme extracellular 595:607 arg1 portion 609:615 the extracellular portion 591:615 the extracellular portion we describe here 591:632 The epidermal growth-factor receptor (EGFR) family is closely related to the IR family and has significant sequence identity to the extracellular portion we describe here. 9690478 9 55 theme central 1226:1232 arg1 site 1234:1237 this central site 1221:1237 this central site 1221:1237 Although the fragment (residues 1-462) does not bind ligand, many of the determinants responsible for hormone binding and ligand specificity map to this central site. 9690478 1 56 theme tyrosine-kinase 213:227 arg1 superfamily 238:248 the tyrosine-kinase receptor superfamily 209:248 the tyrosine-kinase receptor superfamily 209:248 The type-1 insulin-like growth-factor receptor (IGF-1R) and insulin receptor (IR) are closely related members of the tyrosine-kinase receptor superfamily. 20696930 10 0 theme affinity 1520:1527 arg1 maturation 1529:1538 affinity maturation 1520:1538 affinity maturation 1520:1538 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 10 1 theme free 1421:1424 arg1 structures 1442:1451 free and bound Zher2 structures 1421:1451 free and bound Zher2 structures 1421:1451 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 7 2 theme conformational 1023:1036 arg1 interconversion 1038:1052 conformational interconversion 1023:1052 conformational interconversion on a submillisecond time scale 1023:1083 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 7 3 from interconversion 1038:1052 arg1 scale 1079:1083 a submillisecond time scale 1057:1083 a submillisecond time scale 1057:1083 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 1 4 theme breast 227:232 arg1 cancer 234:239 breast cancer 227:239 breast cancer 227:239 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 5 5 dep antibodies 660:669 arg1 antibodies 660:669 the therapeutic antibodies 644:669 the therapeutic antibodies trastuzumab and pertuzumab 644:696 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 5 5 dep antibodies 660:669 arg1 pertuzumab 687:696 pertuzumab 687:696 pertuzumab 687:696 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 5 5 dep antibodies 660:669 arg1 trastuzumab 671:681 trastuzumab 671:681 trastuzumab 671:681 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 10 6 theme bound 1430:1434 arg1 structures 1442:1451 free and bound Zher2 structures 1421:1451 free and bound Zher2 structures 1421:1451 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 6 7 theme diagnostic 777:786 arg1 use 788:790 diagnostic use 777:790 diagnostic use 777:790 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 1 8 theme cancer 234:239 arg1 form 219:222 an aggressive form 205:222 an aggressive form of breast cancer 205:239 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 3 9 theme acid 321:324 arg1 binder 403:408 a high-affinity binder 387:408 a high-affinity binder of HER2 387:416 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 3 9 theme acid 321:324 arg1 residue 326:332 The 58 amino acid residue 308:332 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 10 10 theme original 1391:1398 arg1 scaffold 1409:1416 the original Z domain scaffold 1387:1416 the original Z domain scaffold 1387:1416 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 10 11 theme scaffold 1409:1416 arg1 comparison 1373:1382 A comparison 1371:1382 A comparison of the original Z domain scaffold to free and bound Zher2 structures 1371:1451 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 10 12 theme domain 1402:1407 arg1 scaffold 1409:1416 the original Z domain scaffold 1387:1416 the original Z domain scaffold 1387:1416 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 9 13 theme perfect 1266:1272 arg1 homogeneity 1289:1299 perfect conformational homogeneity 1266:1299 perfect conformational homogeneity 1266:1299 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 5 14 theme conformational 572:585 arg1 epitope 587:593 a conformational epitope 570:593 a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab 570:696 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 4 15 with complex 511:517 arg1 domain 547:552 the HER2 extracellular domain 524:552 the HER2 extracellular domain 524:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 7 16 from stable 981:986 arg1 state 1002:1006 the folded state 991:1006 the folded state 991:1006 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 3 17 theme amino 315:319 arg1 binder 403:408 a high-affinity binder 387:408 a high-affinity binder of HER2 387:416 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 3 17 theme amino 315:319 arg1 residue 326:332 The 58 amino acid residue 308:332 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 4 18 theme Zher2 455:459 arg1 structure 442:450 the structure 438:450 the structure of Zher2 in solution 438:471 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 4 18 theme Zher2 455:459 arg1 structure 489:497 the crystal structure 477:497 the crystal structure of Zher2 in complex with the HER2 extracellular domain 477:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 4 19 theme extracellular 533:545 arg1 domain 547:552 the HER2 extracellular domain 524:552 the HER2 extracellular domain 524:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 10 20 theme Z 1400:1400 arg1 scaffold 1409:1416 the original Z domain scaffold 1387:1416 the original Z domain scaffold 1387:1416 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 10 21 theme surface 1586:1592 arg1 optimization 1594:1605 binding surface optimization 1578:1605 binding surface optimization 1578:1605 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 3 22 theme HER2 413:416 arg1 binder 403:408 a high-affinity binder 387:408 a high-affinity binder of HER2 387:416 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 3 22 theme HER2 413:416 arg1 residue 326:332 The 58 amino acid residue 308:332 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 4 23 from structure 442:450 arg1 complex 511:517 complex 511:517 complex with the HER2 extracellular domain 511:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 4 23 from structure 442:450 arg1 solution 464:471 solution 464:471 solution 464:471 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 7 24 theme Biophysical 914:924 arg1 characterization 926:941 Biophysical characterization 914:941 Biophysical characterization 914:941 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 10 25 theme binding 1578:1584 arg1 optimization 1594:1605 binding surface optimization 1578:1605 binding surface optimization 1578:1605 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 3 26 theme high-affinity 389:401 arg1 binder 403:408 a high-affinity binder 387:408 a high-affinity binder of HER2 387:416 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 3 26 theme high-affinity 389:401 arg1 residue 326:332 The 58 amino acid residue 308:332 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 0 27 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for high-affinity HER2 receptor binding by an engineered protein. 0:81 Structural basis for high-affinity HER2 receptor binding by an engineered protein. 20696930 9 28 theme engineered 1342:1351 arg1 proteins 1361:1368 engineered binding proteins 1342:1368 engineered binding proteins 1342:1368 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 1 29 theme human 87:91 arg1 receptor 117:124 The human epidermal growth factor receptor 2 83:126 The human epidermal growth factor receptor 2 (HER2) 83:133 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 29 theme human 87:91 arg1 HER2 129:132 HER2 129:132 HER2 129:132 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 3 30 theme Zher2 334:338 arg1 molecule 349:356 Zher2 affibody molecule 334:356 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 9 31 with strong 1212:1217 arg1 constant 1239:1246 a dissociation constant 1224:1246 a dissociation constant K(D) = 22 pM 1224:1259 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 5 32 theme therapeutic 648:658 arg1 antibodies 660:669 the therapeutic antibodies 644:669 the therapeutic antibodies trastuzumab and pertuzumab 644:696 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 5 32 theme therapeutic 648:658 arg1 pertuzumab 687:696 pertuzumab 687:696 pertuzumab 687:696 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 5 32 theme therapeutic 648:658 arg1 trastuzumab 671:681 trastuzumab 671:681 trastuzumab 671:681 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 6 33 theme interference 726:737 arg1 lack 718:721 lack 718:721 lack of interference 718:737 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 6 33 theme interference 726:737 arg1 size 709:712 Its small size 699:712 Its small size 699:712 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 1 34 theme epidermal 93:101 arg1 receptor 117:124 The human epidermal growth factor receptor 2 83:126 The human epidermal growth factor receptor 2 (HER2) 83:133 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 34 theme epidermal 93:101 arg1 HER2 129:132 HER2 129:132 HER2 129:132 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 35 theme several 178:184 arg1 form 219:222 an aggressive form 205:222 an aggressive form of breast cancer 205:239 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 35 theme several 178:184 arg1 cancers 186:192 several cancers 178:192 several cancers 178:192 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 2 36 theme cancer 276:281 arg1 diagnostics 283:293 cancer diagnostics 276:293 cancer diagnostics 276:293 It is therefore a target for both cancer diagnostics and therapy. 20696930 9 37 dep = 1253:1253 arg1 pM 1258:1259 22 pM 1255:1259 22 pM 1255:1259 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 10 38 theme high-affinity 1465:1477 arg1 binding 1479:1485 high-affinity binding 1465:1485 high-affinity binding 1465:1485 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 6 39 theme agents 828:833 arg1 delivery 804:811 delivery 804:811 delivery of therapeutic agents to HER2-expressing tumors 804:859 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 1 40 theme growth 103:108 arg1 receptor 117:124 The human epidermal growth factor receptor 2 83:126 The human epidermal growth factor receptor 2 (HER2) 83:133 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 40 theme growth 103:108 arg1 HER2 129:132 HER2 129:132 HER2 129:132 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 41 theme cancers 186:192 arg1 tumors 168:173 tumors 168:173 tumors of several cancers, including an aggressive form of breast cancer 168:239 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 0 42 theme receptor 40:47 arg1 binding 49:55 high-affinity HER2 receptor binding 21:55 high-affinity HER2 receptor binding 21:55 Structural basis for high-affinity HER2 receptor binding by an engineered protein. 20696930 5 43 from epitope 587:593 arg1 HER2 598:601 HER2 598:601 HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab 598:696 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 10 44 dep compromise 1559:1568 arg1 a 1557:1557 a 1557:1557 a 1557:1557 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 6 45 theme therapeutic 816:826 arg1 agents 828:833 therapeutic agents 816:833 therapeutic agents 816:833 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 7 46 from state 1002:1006 arg1 stable 981:986 stable 981:986 stable 981:986 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 0 47 theme HER2 35:38 arg1 binding 49:55 high-affinity HER2 receptor binding 21:55 high-affinity HER2 receptor binding 21:55 Structural basis for high-affinity HER2 receptor binding by an engineered protein. 20696930 4 48 theme HER2 528:531 arg1 domain 547:552 the HER2 extracellular domain 524:552 the HER2 extracellular domain 524:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 9 49 theme dissociation 1226:1237 arg1 constant 1239:1246 a dissociation constant 1224:1246 a dissociation constant K(D) = 22 pM 1224:1259 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 0 50 theme high-affinity 21:33 arg1 binding 49:55 high-affinity HER2 receptor binding 21:55 high-affinity HER2 receptor binding 21:55 Structural basis for high-affinity HER2 receptor binding by an engineered protein. 20696930 4 51 from structure 489:497 arg1 complex 511:517 complex 511:517 complex with the HER2 extracellular domain 511:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 4 51 from structure 489:497 arg1 solution 464:471 solution 464:471 solution 464:471 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 4 52 theme crystal 481:487 arg1 structure 489:497 the crystal structure 477:497 the crystal structure of Zher2 in complex with the HER2 extracellular domain 477:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 8 53 theme HER2-binding 1118:1129 arg1 it 1108:1109 it 1108:1109 it 1108:1109 The data suggest that it is the HER2-binding conformation that is formed transiently prior to binding. 20696930 8 53 theme HER2-binding 1118:1129 arg1 conformation 1131:1142 the HER2-binding conformation 1114:1142 the HER2-binding conformation that is formed transiently prior to binding 1114:1186 The data suggest that it is the HER2-binding conformation that is formed transiently prior to binding. 20696930 6 54 theme HER2-expressing 838:852 arg1 tumors 854:859 HER2-expressing tumors 838:859 HER2-expressing tumors 838:859 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 1 55 theme factor 110:115 arg1 receptor 117:124 The human epidermal growth factor receptor 2 83:126 The human epidermal growth factor receptor 2 (HER2) 83:133 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 1 55 theme factor 110:115 arg1 HER2 129:132 HER2 129:132 HER2 129:132 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 7 56 theme submillisecond 1059:1072 arg1 scale 1079:1083 a submillisecond time scale 1057:1083 a submillisecond time scale 1057:1083 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 4 57 theme Zher2 502:506 arg1 structure 442:450 the structure 438:450 the structure of Zher2 in solution 438:471 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 4 57 theme Zher2 502:506 arg1 structure 489:497 the crystal structure 477:497 the crystal structure of Zher2 in complex with the HER2 extracellular domain 477:552 Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. 20696930 7 58 theme time 1074:1077 arg1 scale 1079:1083 a submillisecond time scale 1057:1083 a submillisecond time scale 1057:1083 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 10 59 theme Zher2 1436:1440 arg1 structures 1442:1451 free and bound Zher2 structures 1421:1451 free and bound Zher2 structures 1421:1451 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 9 60 theme binding 1353:1359 arg1 proteins 1361:1368 engineered binding proteins 1342:1368 engineered binding proteins 1342:1368 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 7 61 theme folded 995:1000 arg1 state 1002:1006 the folded state 991:1006 the folded state 991:1006 Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. 20696930 3 62 dep residue 326:332 arg1 molecule 349:356 Zher2 affibody molecule 334:356 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 6 63 theme small 703:707 arg1 size 709:712 Its small size 699:712 Its small size 699:712 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 1 64 theme aggressive 208:217 arg1 form 219:222 an aggressive form 205:222 an aggressive form of breast cancer 205:239 The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. 20696930 0 65 theme engineered 63:72 arg1 protein 74:80 an engineered protein 60:80 an engineered protein 60:80 Structural basis for high-affinity HER2 receptor binding by an engineered protein. 20696930 10 66 theme unbound 1641:1647 arg1 state 1649:1653 the unbound state 1637:1653 the unbound state 1637:1653 A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached. 20696930 5 67 from those 624:628 arg1 distant 611:617 distant 611:617 distant 611:617 Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. 20696930 6 68 with Zher2 751:755 arg1 advantages 762:771 advantages 762:771 advantages 762:771 Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. 20696930 9 69 theme = 1253:1253 arg1 constant 1239:1246 a dissociation constant 1224:1246 a dissociation constant K(D) = 22 pM 1224:1259 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 20696930 3 70 theme affibody 340:347 arg1 molecule 349:356 Zher2 affibody molecule 334:356 The 58 amino acid residue Zher2 affibody molecule 308:356 The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. 20696930 9 71 theme conformational 1274:1287 arg1 homogeneity 1289:1299 perfect conformational homogeneity 1266:1299 perfect conformational homogeneity 1266:1299 Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. 12646248 2 0 theme selective 302:310 arg1 inhibitors 312:321 DPPIV selective inhibitors 296:321 DPPIV selective inhibitors 296:321 Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. 12646248 4 1 theme beta-propeller 516:529 arg1 domain 531:536 a unique eight-bladed beta-propeller domain 494:536 a unique eight-bladed beta-propeller domain in the N-terminal region 494:561 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 6 2 theme sequence 795:802 arg1 Comparison 758:767 Comparison 758:767 Comparison of the overall amino acid sequence between human DPPIV and POP 758:830 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 3 3 theme DPPIV 443:447 arg1 structure 424:432 the crystal structure 412:432 the crystal structure of human DPPIV at 2.6A resolution 412:466 We determined the crystal structure of human DPPIV at 2.6A resolution. 12646248 1 4 theme family 213:218 arg1 protease 159:166 a serine protease 150:166 a serine protease 150:166 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 1 4 theme family 213:218 arg1 member 171:176 a member 169:176 a member of the prolyl oligopeptidase (POP) family 169:218 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 0 5 theme eight-bladed 81:92 arg1 fold 109:112 a unique eight-bladed beta-propeller fold 72:112 a unique eight-bladed beta-propeller fold 72:112 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 6 6 theme acid 790:793 arg1 sequence 795:802 the overall amino acid sequence 772:802 the overall amino acid sequence 772:802 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 7 7 theme eight-bladed 947:958 arg1 domain 975:980 a unique eight-bladed beta-propeller domain 938:980 a unique eight-bladed beta-propeller domain 938:980 In this paper, we report the structure of human DPPIV, especially focusing on a unique eight-bladed beta-propeller domain. 12646248 0 8 theme unique 74:79 arg1 fold 109:112 a unique eight-bladed beta-propeller fold 72:112 a unique eight-bladed beta-propeller fold 72:112 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 6 9 theme amino 784:788 arg1 sequence 795:802 the overall amino acid sequence 772:802 the overall amino acid sequence 772:802 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 7 10 theme beta-propeller 960:973 arg1 domain 975:980 a unique eight-bladed beta-propeller domain 938:980 a unique eight-bladed beta-propeller domain 938:980 In this paper, we report the structure of human DPPIV, especially focusing on a unique eight-bladed beta-propeller domain. 12646248 3 11 theme human 437:441 arg1 DPPIV 443:447 human DPPIV 437:447 human DPPIV 437:447 We determined the crystal structure of human DPPIV at 2.6A resolution. 12646248 6 12 theme overall 776:782 arg1 sequence 795:802 the overall amino acid sequence 772:802 the overall amino acid sequence 772:802 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 5 13 located found 712:716 arg2 structure 641:649 the large "cave" structure 624:649 the large "cave" structure 624:649 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 5 13 located found 712:716 arg1 side 725:728 the side 721:728 the side of the beta-propeller fold 721:755 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 5 14 theme beta-propeller 737:750 arg1 fold 752:755 the beta-propeller fold 733:755 the beta-propeller fold 733:755 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 0 15 theme beta-propeller 94:107 arg1 fold 109:112 a unique eight-bladed beta-propeller fold 72:112 a unique eight-bladed beta-propeller fold 72:112 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 7 16 theme unique 940:945 arg1 domain 975:980 a unique eight-bladed beta-propeller domain 938:980 a unique eight-bladed beta-propeller domain 938:980 In this paper, we report the structure of human DPPIV, especially focusing on a unique eight-bladed beta-propeller domain. 12646248 1 17 theme serine 152:157 arg1 protease 159:166 a serine protease 150:166 a serine protease 150:166 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 1 17 theme serine 152:157 arg1 member 171:176 a member 169:176 a member of the prolyl oligopeptidase (POP) family 169:218 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 1 17 theme serine 152:157 arg1 peptidase 126:134 Dipeptidyl peptidase IV 115:137 Dipeptidyl peptidase IV (DPPIV) 115:145 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 7 18 theme human 902:906 arg1 DPPIV 908:912 human DPPIV 902:912 human DPPIV 902:912 In this paper, we report the structure of human DPPIV, especially focusing on a unique eight-bladed beta-propeller domain. 12646248 4 19 from domain 585:590 arg1 region 556:561 the N-terminal region 541:561 the N-terminal region 541:561 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 4 19 from domain 585:590 arg1 region 610:615 the C-terminal region 595:615 the C-terminal region 595:615 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 4 20 theme C-terminal 599:608 arg1 region 610:615 the C-terminal region 595:615 the C-terminal region 595:615 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 2 21 theme DPPIV 377:381 arg1 function 365:372 the biological function 350:372 the biological function of DPPIV 350:381 Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. 12646248 2 22 theme inhibitors 312:321 arg1 development 281:291 the development 277:291 the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, 277:382 Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. 12646248 2 22 theme inhibitors 312:321 arg1 important 387:395 important 387:395 important 387:395 Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. 12646248 5 23 theme large 628:632 arg1 structure 641:649 the large "cave" structure 624:649 the large "cave" structure 624:649 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 4 24 theme serine 569:574 arg1 domain 585:590 a serine protease domain 567:590 a serine protease domain in the C-terminal region 567:615 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 8 25 theme substrate 1029:1037 arg1 access 1015:1020 the access 1011:1020 the access of the substrate to this domain 1011:1052 We also discuss the way for the access of the substrate to this domain. 12646248 4 26 theme unique 496:501 arg1 domain 531:536 a unique eight-bladed beta-propeller domain 494:536 a unique eight-bladed beta-propeller domain in the N-terminal region 494:561 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 4 27 theme N-terminal 545:554 arg1 region 556:561 the N-terminal region 541:561 the N-terminal region 541:561 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 2 28 theme DPPIV 296:300 arg1 inhibitors 312:321 DPPIV selective inhibitors 296:321 DPPIV selective inhibitors 296:321 Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. 12646248 2 29 theme biological 354:363 arg1 function 365:372 the biological function 350:372 the biological function of DPPIV 350:381 Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. 12646248 0 30 theme human 30:34 arg1 peptidase 47:55 human dipeptidyl peptidase IV 30:58 human dipeptidyl peptidase IV 30:58 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 6 31 theme low 838:840 arg1 homology 842:849 low homology 838:849 low homology (12.9%) 838:857 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 6 31 theme low 838:840 arg1 % 856:856 12.9% 852:856 12.9% 852:856 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 5 32 theme cave 635:638 arg1 structure 641:649 the large "cave" structure 624:649 the large "cave" structure 624:649 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 5 33 theme " 639:639 arg1 structure 641:649 the large "cave" structure 624:649 the large "cave" structure 624:649 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 1 34 theme several 248:254 arg1 diseases 256:263 several diseases 248:263 several diseases 248:263 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 4 35 theme eight-bladed 503:514 arg1 domain 531:536 a unique eight-bladed beta-propeller domain 494:536 a unique eight-bladed beta-propeller domain in the N-terminal region 494:561 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 4 36 from domain 531:536 arg1 region 556:561 the N-terminal region 541:561 the N-terminal region 541:561 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 4 36 from domain 531:536 arg1 region 610:615 the C-terminal region 595:615 the C-terminal region 595:615 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 4 37 theme protease 576:583 arg1 domain 585:590 a serine protease domain 567:590 a serine protease domain in the C-terminal region 567:615 The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. 12646248 0 38 theme peptidase 47:55 arg1 function 18:25 function 18:25 function 18:25 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 0 38 theme peptidase 47:55 arg1 structure 4:12 structure 4:12 structure 4:12 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 5 39 theme substrate 698:706 arg1 access 684:689 the access 680:689 the access of the substrate 680:706 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 7 40 theme DPPIV 908:912 arg1 structure 889:897 the structure 885:897 the structure of human DPPIV 885:912 In this paper, we report the structure of human DPPIV, especially focusing on a unique eight-bladed beta-propeller domain. 12646248 0 41 theme dipeptidyl 36:45 arg1 peptidase 47:55 human dipeptidyl peptidase IV 30:58 human dipeptidyl peptidase IV 30:58 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 1 42 theme Dipeptidyl 115:124 arg1 peptidase 126:134 Dipeptidyl peptidase IV 115:137 Dipeptidyl peptidase IV (DPPIV) 115:145 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 1 42 theme Dipeptidyl 115:124 arg1 DPPIV 140:144 DPPIV 140:144 DPPIV 140:144 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 1 42 theme Dipeptidyl 115:124 arg1 protease 159:166 a serine protease 150:166 a serine protease 150:166 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 1 43 theme prolyl 185:190 arg1 family 213:218 the prolyl oligopeptidase (POP) family 181:218 the prolyl oligopeptidase (POP) family 181:218 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 3 44 theme crystal 416:422 arg1 structure 424:432 the crystal structure 412:432 the crystal structure of human DPPIV at 2.6A resolution 412:466 We determined the crystal structure of human DPPIV at 2.6A resolution. 12646248 1 45 theme oligopeptidase 192:205 arg1 family 213:218 the prolyl oligopeptidase (POP) family 181:218 the prolyl oligopeptidase (POP) family 181:218 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 12646248 0 46 dep structure 4:12 arg1 The 0:2 The 0:2 The 0:2 The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. 12646248 5 47 theme fold 752:755 arg1 side 725:728 the side 721:728 the side of the beta-propeller fold 721:755 Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta-propeller fold. 12646248 6 48 theme human 812:816 arg1 DPPIV 818:822 human DPPIV 812:822 human DPPIV 812:822 Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). 12646248 3 49 theme 2.6A 452:455 arg1 resolution 457:466 2.6A resolution 452:466 2.6A resolution 452:466 We determined the crystal structure of human DPPIV at 2.6A resolution. 12646248 3 50 from resolution 457:466 arg1 structure 424:432 the crystal structure 412:432 the crystal structure of human DPPIV at 2.6A resolution 412:466 We determined the crystal structure of human DPPIV at 2.6A resolution. 12646248 1 51 theme POP 208:210 arg1 family 213:218 the prolyl oligopeptidase (POP) family 181:218 the prolyl oligopeptidase (POP) family 181:218 Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. 11772019 0 0 theme human 57:61 arg1 alpha-amylase 74:86 human pancreatic alpha-amylase 57:86 human pancreatic alpha-amylase 57:86 Probing the role of the chloride ion in the mechanism of human pancreatic alpha-amylase. 11772019 0 1 from role 12:15 arg1 mechanism 44:52 the mechanism 40:52 the mechanism of human pancreatic alpha-amylase 40:86 Probing the role of the chloride ion in the mechanism of human pancreatic alpha-amylase. 11772019 5 2 theme X-ray 640:644 arg1 analysis 663:670 X-ray crystallographic analysis 640:670 X-ray crystallographic analysis 640:670 X-ray crystallographic analysis revealed that these mutations did not result in significant structural changes. 11772019 8 3 theme lower 1266:1270 arg1 affinity 1280:1287 much lower binding affinity 1261:1287 much lower binding affinity for chloride but still required chloride for maximal activity 1261:1349 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 5 4 theme crystallographic 646:661 arg1 analysis 663:670 X-ray crystallographic analysis 640:670 X-ray crystallographic analysis 640:670 X-ray crystallographic analysis revealed that these mutations did not result in significant structural changes. 11772019 0 5 theme alpha-amylase 74:86 arg1 mechanism 44:52 the mechanism 40:52 the mechanism of human pancreatic alpha-amylase 40:86 Probing the role of the chloride ion in the mechanism of human pancreatic alpha-amylase. 11772019 8 6 theme maximal 1334:1340 arg1 activity 1342:1349 maximal activity 1334:1349 maximal activity 1334:1349 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 3 7 theme ion 443:445 arg1 site 455:458 the chloride ion binding site 430:458 the chloride ion binding site 430:458 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 4 8 theme R337 619:622 arg1 enzymes 631:637 the R195 and R337 mutant enzymes 606:637 enzymes 631:637 Mutations in this binding site were found to severely affect the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes. 11772019 4 9 theme binding 493:499 arg1 site 501:504 this binding site 488:504 this binding site 488:504 Mutations in this binding site were found to severely affect the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes. 11772019 0 10 theme pancreatic 63:72 arg1 alpha-amylase 74:86 human pancreatic alpha-amylase 57:86 human pancreatic alpha-amylase 57:86 Probing the role of the chloride ion in the mechanism of human pancreatic alpha-amylase. 11772019 6 11 theme mutations 787:795 arg1 introduction 765:776 the introduction 761:776 the introduction of these mutations 761:795 However, the introduction of these mutations did alter the kinetic properties of the enzyme. 11772019 3 12 from residues 418:425 arg1 site 455:458 the chloride ion binding site 430:458 the chloride ion binding site 430:458 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 3 13 theme binding 447:453 arg1 site 455:458 the chloride ion binding site 430:458 the chloride ion binding site 430:458 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 7 14 theme amino 1048:1052 arg1 acid 1054:1057 a nonbasic amino acid 1037:1057 a nonbasic amino acid 1037:1057 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 7 15 from decrease 897:904 arg1 activity 913:920 the activity 909:920 the activity of the enzyme toward starch 909:948 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 7 16 with replacement 1012:1022 arg1 acid 1054:1057 a nonbasic amino acid 1037:1057 a nonbasic amino acid 1037:1057 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 4 17 theme HPA 551:553 arg1 ability 540:546 the ability 536:546 the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes 536:637 Mutations in this binding site were found to severely affect the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes. 11772019 0 18 theme ion 33:35 arg1 role 12:15 the role 8:15 the role of the chloride ion in the mechanism of human pancreatic alpha-amylase 8:86 Probing the role of the chloride ion in the mechanism of human pancreatic alpha-amylase. 11772019 3 19 dep mutants 351:357 arg1 R337A 381:385 R337A 381:385 R337A 381:385 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 3 19 dep mutants 351:357 arg1 R337Q 392:396 R337Q 392:396 R337Q 392:396 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 3 19 dep mutants 351:357 arg1 N298S 374:378 N298S 374:378 N298S 374:378 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 3 19 dep mutants 351:357 arg1 R195Q 367:371 R195Q 367:371 R195Q 367:371 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 3 19 dep mutants 351:357 arg1 mutants 351:357 mutants 351:357 mutants (R195A, R195Q, N298S, R337A, and R337Q) 351:397 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 3 20 theme chloride 434:441 arg1 site 455:458 the chloride ion binding site 430:458 the chloride ion binding site 430:458 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 8 21 contain had 1257:1259 arg1 enzyme 1245:1250 an enzyme 1242:1250 an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity 1242:1349 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 8 21 contain had 1257:1259 arg2 affinity 1280:1287 much lower binding affinity 1261:1287 much lower binding affinity for chloride but still required chloride for maximal activity 1261:1349 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 7 22 contain has 1139:1141 arg1 alpha-amylase 1074:1086 an alpha-amylase 1071:1086 an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA 1071:1187 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 7 22 contain has 1139:1141 arg2 profile 1148:1154 a pH profile 1143:1154 a pH profile similar to that of wild-type HPA 1143:1187 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 2 23 theme maximal 270:276 arg1 activity 278:285 maximal activity 270:285 maximal activity 270:285 Some members of this family, including HPA, require chloride for maximal activity. 11772019 1 24 theme alpha-amylase 145:157 arg1 family 159:164 the alpha-amylase family 141:164 the alpha-amylase family involved in the degradation of starch 141:202 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 7 25 theme nonbasic 1039:1046 arg1 acid 1054:1057 a nonbasic amino acid 1037:1057 a nonbasic amino acid 1037:1057 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 7 26 theme pH 966:967 arg1 optimum 969:975 the pH optimum 962:975 the pH optimum 962:975 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 1 27 theme family 159:164 arg1 alpha-amylase 106:118 Human pancreatic alpha-amylase 89:118 Human pancreatic alpha-amylase (HPA) 89:124 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 1 27 theme family 159:164 arg1 member 131:136 a member 129:136 a member of the alpha-amylase family involved in the degradation of starch 129:202 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 3 28 theme activation 327:336 arg1 mechanism 305:313 the mechanism 301:313 the mechanism of chloride activation 301:336 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 4 29 theme mutant 624:629 arg1 enzymes 631:637 the R195 and R337 mutant enzymes 606:637 enzymes 631:637 Mutations in this binding site were found to severely affect the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes. 11772019 7 30 theme similar 1156:1162 arg1 profile 1148:1154 a pH profile 1143:1154 a pH profile similar to that of wild-type HPA 1143:1187 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 8 31 theme binding 1272:1278 arg1 affinity 1280:1287 much lower binding affinity 1261:1287 much lower binding affinity for chloride but still required chloride for maximal activity 1261:1349 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 1 32 theme Human 89:93 arg1 HPA 121:123 HPA 121:123 HPA 121:123 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 1 32 theme Human 89:93 arg1 alpha-amylase 106:118 Human pancreatic alpha-amylase 89:118 Human pancreatic alpha-amylase (HPA) 89:124 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 1 32 theme Human 89:93 arg1 member 131:136 a member 129:136 a member of the alpha-amylase family involved in the degradation of starch 129:202 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 5 33 theme significant 720:730 arg1 changes 743:749 significant structural changes 720:749 significant structural changes 720:749 X-ray crystallographic analysis revealed that these mutations did not result in significant structural changes. 11772019 0 34 theme chloride 24:31 arg1 ion 33:35 the chloride ion 20:35 the chloride ion 20:35 Probing the role of the chloride ion in the mechanism of human pancreatic alpha-amylase. 11772019 7 35 theme pH 1145:1146 arg1 profile 1148:1154 a pH profile 1143:1154 a pH profile similar to that of wild-type HPA 1143:1187 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 5 36 theme structural 732:741 arg1 changes 743:749 significant structural changes 720:749 significant structural changes 720:749 X-ray crystallographic analysis revealed that these mutations did not result in significant structural changes. 11772019 7 37 theme wild-type 1175:1183 arg1 HPA 1185:1187 wild-type HPA 1175:1187 wild-type HPA 1175:1187 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 6 38 theme enzyme 837:842 arg1 properties 819:828 the kinetic properties 807:828 the kinetic properties of the enzyme 807:842 However, the introduction of these mutations did alter the kinetic properties of the enzyme. 11772019 7 39 theme R337 1027:1030 arg1 replacement 1012:1022 replacement 1012:1022 replacement of R337 with a nonbasic amino acid 1012:1057 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 4 40 from Mutations 475:483 arg1 site 501:504 this binding site 488:504 this binding site 488:504 Mutations in this binding site were found to severely affect the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes. 11772019 9 41 theme R337 1502:1505 arg1 presence 1490:1497 the presence 1486:1497 the presence of R337, a positively charged residue 1486:1535 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 1 42 theme pancreatic 95:104 arg1 HPA 121:123 HPA 121:123 HPA 121:123 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 1 42 theme pancreatic 95:104 arg1 alpha-amylase 106:118 Human pancreatic alpha-amylase 89:118 Human pancreatic alpha-amylase (HPA) 89:124 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 1 42 theme pancreatic 95:104 arg1 member 131:136 a member 129:136 a member of the alpha-amylase family involved in the degradation of starch 129:202 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 3 43 theme mutants 351:357 arg1 series 341:346 a series 339:346 a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) 339:397 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 11772019 7 44 theme enzyme 929:934 arg1 activity 913:920 the activity 909:920 the activity of the enzyme toward starch 909:948 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 7 45 theme 20-450-fold 885:895 arg1 decrease 897:904 a 20-450-fold decrease 883:904 a 20-450-fold decrease in the activity of the enzyme toward starch 883:948 Mutations to residue R195 resulted in a 20-450-fold decrease in the activity of the enzyme toward starch and shifted the pH optimum to a more basic pH. Interestingly, replacement of R337 with a nonbasic amino acid resulted in an alpha-amylase that no longer required chloride for catalysis and has a pH profile similar to that of wild-type HPA. 11772019 2 46 theme family 226:231 arg1 HPA 244:246 HPA 244:246 HPA 244:246 Some members of this family, including HPA, require chloride for maximal activity. 11772019 2 46 theme family 226:231 arg1 members 210:216 Some members 205:216 Some members 205:216 Some members of this family, including HPA, require chloride for maximal activity. 11772019 8 47 theme required 1312:1319 arg1 chloride 1321:1328 still required chloride 1306:1328 still required chloride 1306:1328 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 9 48 theme acid/base 1422:1430 arg1 E233 1442:1445 E233 1442:1445 E233 1442:1445 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 9 48 theme acid/base 1422:1430 arg1 catalyst 1432:1439 the acid/base catalyst 1418:1439 the acid/base catalyst 1418:1439 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 9 48 theme acid/base 1422:1430 arg1 due 1479:1481 due 1479:1481 due 1479:1481 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 1 49 theme starch 197:202 arg1 degradation 182:192 the degradation 178:192 the degradation of starch 178:202 Human pancreatic alpha-amylase (HPA) is a member of the alpha-amylase family involved in the degradation of starch. 11772019 9 50 theme charged 1521:1527 arg1 residue 1529:1535 a positively charged residue 1508:1535 a positively charged residue 1508:1535 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 9 50 theme charged 1521:1527 arg1 R337 1502:1505 R337 1502:1505 R337 1502:1505 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 9 51 theme catalyst 1432:1439 arg1 a 1412:1412 a 1412:1412 a 1412:1412 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 9 51 theme catalyst 1432:1439 arg1 pK 1409:1410 the pK 1405:1410 the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue 1405:1535 We propose that the chloride is required to increase the pK(a) of the acid/base catalyst, E233, which would otherwise be lower due to the presence of R337, a positively charged residue. 11772019 6 52 theme kinetic 811:817 arg1 properties 819:828 the kinetic properties 807:828 the kinetic properties of the enzyme 807:842 However, the introduction of these mutations did alter the kinetic properties of the enzyme. 11772019 4 53 theme chloride 563:570 arg1 ions 572:575 chloride ions 563:575 chloride ions 563:575 Mutations in this binding site were found to severely affect the ability of HPA to bind chloride ions with no binding detected for the R195 and R337 mutant enzymes. 11772019 8 54 from residue 1217:1223 arg1 mutation 1205:1212 a mutation 1203:1212 a mutation at residue N298 1203:1228 In contrast, a mutation at residue N298 resulted in an enzyme that had much lower binding affinity for chloride but still required chloride for maximal activity. 11772019 3 55 theme chloride 318:325 arg1 activation 327:336 chloride activation 318:336 chloride activation 318:336 To determine the mechanism of chloride activation, a series of mutants (R195A, R195Q, N298S, R337A, and R337Q) were made in which residues in the chloride ion binding site were replaced. 8978667 4 0 theme domain 435:440 arg1 part 410:413 part 410:413 part of this interacting domain 410:440 Cysteine 69 which is part of this interacting domain is implicated in the formation of the disulfide bond. 8978667 5 1 theme K+ 576:577 arg1 expression 587:596 functional K+ channel expression 565:596 functional K+ channel expression 565:596 Replacing this cysteine with a serine residue results in the loss of functional K+ channel expression. 8978667 6 2 theme association 639:649 arg1 This 599:602 This 599:602 This 599:602 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 6 2 theme association 639:649 arg1 example 617:623 the first example 607:623 the first example of a covalent association of functional subunits in voltage-sensitive channels 607:702 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 5 3 theme channel 579:585 arg1 expression 587:596 functional K+ channel expression 565:596 functional K+ channel expression 565:596 Replacing this cysteine with a serine residue results in the loss of functional K+ channel expression. 8978667 5 4 theme serine 527:532 arg1 residue 534:540 a serine residue 525:540 a serine residue 525:540 Replacing this cysteine with a serine residue results in the loss of functional K+ channel expression. 8978667 3 5 theme 34 311:312 arg1 domain 325:330 a 34 amino acid domain 309:330 a 34 amino acid domain that is localized to the extracellular M1P1 linker loop 309:386 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 3 5 theme 34 311:312 arg1 localized 340:348 localized 340:348 localized 340:348 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 5 6 theme expression 587:596 arg1 loss 557:560 the loss 553:560 the loss of functional K+ channel expression 553:596 Replacing this cysteine with a serine residue results in the loss of functional K+ channel expression. 8978667 3 7 theme amino 314:318 arg1 domain 325:330 a 34 amino acid domain 309:330 a 34 amino acid domain that is localized to the extracellular M1P1 linker loop 309:386 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 3 7 theme amino 314:318 arg1 localized 340:348 localized 340:348 localized 340:348 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 6 8 from example 617:623 arg1 channels 695:702 voltage-sensitive channels 677:702 voltage-sensitive channels 677:702 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 6 9 theme covalent 630:637 arg1 association 639:649 a covalent association 628:649 a covalent association of functional subunits 628:672 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 4 10 theme interacting 423:433 arg1 domain 435:440 this interacting domain 418:440 this interacting domain 418:440 Cysteine 69 which is part of this interacting domain is implicated in the formation of the disulfide bond. 8978667 2 11 theme disulfide 268:276 arg1 bridge 278:283 an interchain disulfide bridge 254:283 an interchain disulfide bridge 254:283 Here we show that TWIK-1 subunits can self-associate to give dimers containing an interchain disulfide bridge. 8978667 3 12 theme M1P1 371:374 arg1 loop 383:386 the extracellular M1P1 linker loop 353:386 the extracellular M1P1 linker loop 353:386 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 1 13 with type 83:86 arg1 domains 113:119 two P domains 107:119 two P domains 107:119 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 4 14 theme bond 490:493 arg1 formation 463:471 the formation 459:471 the formation of the disulfide bond 459:493 Cysteine 69 which is part of this interacting domain is implicated in the formation of the disulfide bond. 8978667 5 15 theme functional 565:574 arg1 expression 587:596 functional K+ channel expression 565:596 functional K+ channel expression 565:596 Replacing this cysteine with a serine residue results in the loss of functional K+ channel expression. 8978667 6 16 theme disulfide 710:718 arg1 bridge 720:725 a disulfide bridge 708:725 a disulfide bridge 708:725 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 2 17 theme interchain 257:266 arg1 bridge 278:283 an interchain disulfide bridge 254:283 an interchain disulfide bridge 254:283 Here we show that TWIK-1 subunits can self-associate to give dimers containing an interchain disulfide bridge. 8978667 6 18 theme first 611:615 arg1 This 599:602 This 599:602 This 599:602 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 6 18 theme first 611:615 arg1 example 617:623 the first example 607:623 the first example of a covalent association of functional subunits in voltage-sensitive channels 607:702 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 3 19 theme linker 376:381 arg1 loop 383:386 the extracellular M1P1 linker loop 353:386 the extracellular M1P1 linker loop 353:386 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 3 20 theme acid 320:323 arg1 domain 325:330 a 34 amino acid domain 309:330 a 34 amino acid domain that is localized to the extracellular M1P1 linker loop 309:386 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 3 20 theme acid 320:323 arg1 localized 340:348 localized 340:348 localized 340:348 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 4 21 theme disulfide 480:488 arg1 bond 490:493 the disulfide bond 476:493 the disulfide bond 476:493 Cysteine 69 which is part of this interacting domain is implicated in the formation of the disulfide bond. 8978667 1 22 theme new 79:81 arg1 TWIK-1 67:72 TWIK-1 67:72 TWIK-1 67:72 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 1 22 theme new 79:81 arg1 type 83:86 a new type 77:86 a new type of K+ channel with two P domains 77:119 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 1 23 theme human 152:156 arg1 heart 158:162 heart 158:162 heart 158:162 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 0 24 theme TWIK-1 16:21 arg1 subunits 34:41 TWIK-1 K+ channel subunits 16:41 TWIK-1 K+ channel subunits 16:41 Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge. 8978667 0 25 theme subunits 34:41 arg1 Dimerization 0:11 Dimerization 0:11 Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge. 0:65 Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge. 8978667 0 26 theme channel 26:32 arg1 subunits 34:41 TWIK-1 K+ channel subunits 16:41 TWIK-1 K+ channel subunits 16:41 Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge. 8978667 2 27 contain containing 243:252 arg2 bridge 278:283 an interchain disulfide bridge 254:283 an interchain disulfide bridge 254:283 Here we show that TWIK-1 subunits can self-associate to give dimers containing an interchain disulfide bridge. 8978667 2 27 contain containing 243:252 arg1 dimers 236:241 dimers 236:241 dimers containing an interchain disulfide bridge 236:283 Here we show that TWIK-1 subunits can self-associate to give dimers containing an interchain disulfide bridge. 8978667 3 28 theme extracellular 357:369 arg1 loop 383:386 the extracellular M1P1 linker loop 353:386 the extracellular M1P1 linker loop 353:386 This assembly involves a 34 amino acid domain that is localized to the extracellular M1P1 linker loop. 8978667 0 29 theme K+ 23:24 arg1 subunits 34:41 TWIK-1 K+ channel subunits 16:41 TWIK-1 K+ channel subunits 16:41 Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge. 8978667 6 30 theme voltage-sensitive 677:693 arg1 channels 695:702 voltage-sensitive channels 677:702 voltage-sensitive channels 677:702 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 1 31 theme K+ 91:92 arg1 channel 94:100 K+ channel 91:100 K+ channel 91:100 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 1 32 theme channel 94:100 arg1 TWIK-1 67:72 TWIK-1 67:72 TWIK-1 67:72 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 1 32 theme channel 94:100 arg1 type 83:86 a new type 77:86 a new type of K+ channel with two P domains 77:119 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 8978667 6 33 theme subunits 665:672 arg1 association 639:649 a covalent association 628:649 a covalent association of functional subunits 628:672 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 2 34 theme TWIK-1 193:198 arg1 subunits 200:207 TWIK-1 subunits 193:207 TWIK-1 subunits 193:207 Here we show that TWIK-1 subunits can self-associate to give dimers containing an interchain disulfide bridge. 8978667 6 35 theme functional 654:663 arg1 subunits 665:672 functional subunits 654:672 functional subunits 654:672 This is the first example of a covalent association of functional subunits in voltage-sensitive channels via a disulfide bridge. 8978667 0 36 theme disulfide 49:57 arg1 bridge 59:64 a disulfide bridge 47:64 a disulfide bridge 47:64 Dimerization of TWIK-1 K+ channel subunits via a disulfide bridge. 8978667 1 37 theme P 111:111 arg1 domains 113:119 two P domains 107:119 two P domains 107:119 TWIK-1 is a new type of K+ channel with two P domains and is abundantly expressed in human heart and brain. 27418511 5 0 theme -serine-dependent 785:801 arg1 GluD2 803:807 -serine-dependent GluD2 785:807 D: -serine-dependent GluD2 signaling 782:817 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 1 1 theme supramolecular 161:174 arg1 complexes 176:184 supramolecular complexes 161:184 supramolecular complexes that modulate their location and function at excitatory synapses 161:249 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 6 2 theme small-molecule 1016:1029 arg1 ligands 1031:1037 small-molecule ligands 1016:1037 small-molecule ligands 1016:1037 These results lead to a model where protein and small-molecule ligands synergistically control synaptic iGluR function. 27418511 2 3 theme isolated 301:308 arg1 receptors 310:318 isolated receptors 301:318 isolated receptors 301:318 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 27418511 5 4 theme coordination 935:946 arg1 depression 854:863 long-term depression 844:863 long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice 844:965 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 2 5 theme information 282:292 arg1 lack 263:266 a lack 261:266 a lack of structural information beyond isolated receptors or fragments thereof 261:339 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 27418511 4 6 theme domain 685:690 arg1 dimers 692:697 amino-terminal domain dimers 670:697 amino-terminal domain dimers 670:697 We show how Cbln1 hexamers "anchor" GluD2 amino-terminal domain dimers to monomeric β-NRX1. 27418511 5 7 theme fiber-Purkinje 888:901 arg1 synapses 916:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 5 8 theme motor 929:933 arg1 coordination 935:946 motor coordination 929:946 motor coordination 929:946 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 0 9 theme GluD 36:39 arg1 receptors 41:49 GluD receptors 36:49 GluD receptors 36:49 Structural basis for integration of GluD receptors within synaptic organizer complexes. 27418511 2 10 theme structural 271:280 arg1 information 282:292 structural information 271:292 structural information beyond isolated receptors or fragments thereof 271:339 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 27418511 4 11 theme " 662:662 arg1 hexamers 646:653 Cbln1 hexamers 640:653 Cbln1 hexamers "anchor" 640:662 We show how Cbln1 hexamers "anchor" GluD2 amino-terminal domain dimers to monomeric β-NRX1. 27418511 3 12 theme presynaptic 552:562 arg1 β-neurexin 564:573 presynaptic β-neurexin 1 552:575 presynaptic β-neurexin 1 (β-NRX1) 552:584 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 3 12 theme presynaptic 552:562 arg1 β-NRX1 578:583 β-NRX1 578:583 β-NRX1 578:583 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 1 13 theme Ionotropic 88:97 arg1 iGluR 119:123 iGluR 119:123 iGluR 119:123 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 1 13 theme Ionotropic 88:97 arg1 receptor 109:116 Ionotropic glutamate receptor 88:116 Ionotropic glutamate receptor (iGluR) family members 88:139 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 3 14 theme functional 453:462 arg1 analyses 464:471 structural and functional analyses 438:471 structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer 438:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 0 15 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for integration of GluD receptors within synaptic organizer complexes 0:85 Structural basis for integration of GluD receptors within synaptic organizer complexes. 27418511 1 16 theme glutamate 99:107 arg1 iGluR 119:123 iGluR 119:123 iGluR 119:123 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 1 16 theme glutamate 99:107 arg1 receptor 109:116 Ionotropic glutamate receptor 88:116 Ionotropic glutamate receptor (iGluR) family members 88:139 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 2 17 theme signaling 411:419 arg1 understanding 374:386 the mechanistic understanding 358:386 the mechanistic understanding of physiological iGluR signaling 358:419 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 27418511 1 18 theme receptor 109:116 arg1 members 133:139 Ionotropic glutamate receptor (iGluR) family members 88:139 Ionotropic glutamate receptor (iGluR) family members 88:139 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 3 19 theme bridge 503:508 arg1 analyses 464:471 structural and functional analyses 438:471 structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer 438:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 5 20 theme long-term 844:852 arg1 depression 854:863 long-term depression 844:863 long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice 844:965 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 4 21 theme monomeric 702:710 arg1 β-NRX1 712:717 monomeric β-NRX1 702:717 monomeric β-NRX1 702:717 We show how Cbln1 hexamers "anchor" GluD2 amino-terminal domain dimers to monomeric β-NRX1. 27418511 4 22 theme Cbln1 640:644 arg1 hexamers 646:653 Cbln1 hexamers 640:653 Cbln1 hexamers "anchor" 640:662 We show how Cbln1 hexamers "anchor" GluD2 amino-terminal domain dimers to monomeric β-NRX1. 27418511 3 23 theme structural 438:447 arg1 analyses 464:471 structural and functional analyses 438:471 structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer 438:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 5 24 theme cerebellar 868:877 arg1 synapses 916:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 3 25 theme synaptic 608:615 arg1 organizer 617:625 a C1q-like synaptic organizer 597:625 a C1q-like synaptic organizer 597:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 3 25 theme synaptic 608:615 arg1 Cbln1 590:594 Cbln1 590:594 Cbln1 590:594 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 4 26 theme amino-terminal 670:683 arg1 dimers 692:697 amino-terminal domain dimers 670:697 amino-terminal domain dimers 670:697 We show how Cbln1 hexamers "anchor" GluD2 amino-terminal domain dimers to monomeric β-NRX1. 27418511 3 27 theme prototypical 480:491 arg1 bridge 503:508 the prototypical molecular bridge 476:508 the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer 476:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 6 28 theme iGluR 1072:1076 arg1 function 1078:1085 synaptic iGluR function 1063:1085 synaptic iGluR function 1063:1085 These results lead to a model where protein and small-molecule ligands synergistically control synaptic iGluR function. 27418511 5 29 dep D 782:782 arg1 GluD2 803:807 -serine-dependent GluD2 785:807 D: -serine-dependent GluD2 signaling 782:817 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 5 30 theme parallel 879:886 arg1 synapses 916:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 3 31 theme molecular 493:501 arg1 bridge 503:508 the prototypical molecular bridge 476:508 the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer 476:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 5 32 theme D 782:782 arg1 signaling 809:817 D: -serine-dependent GluD2 signaling 782:817 D: -serine-dependent GluD2 signaling 782:817 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 2 33 theme mechanistic 362:372 arg1 understanding 374:386 the mechanistic understanding 358:386 the mechanistic understanding of physiological iGluR signaling 358:419 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 27418511 0 34 theme receptors 41:49 arg1 integration 21:31 integration 21:31 integration of GluD receptors within synaptic organizer complexes 21:85 Structural basis for integration of GluD receptors within synaptic organizer complexes. 27418511 3 35 theme C1q-like 599:606 arg1 organizer 617:625 a C1q-like synaptic organizer 597:625 a C1q-like synaptic organizer 597:625 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 3 35 theme C1q-like 599:606 arg1 Cbln1 590:594 Cbln1 590:594 Cbln1 590:594 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 5 36 from depression 854:863 arg1 mice 962:965 developing mice 951:965 developing mice 951:965 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 5 37 theme developing 951:960 arg1 mice 962:965 developing mice 951:965 developing mice 951:965 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 6 38 theme synaptic 1063:1070 arg1 function 1078:1085 synaptic iGluR function 1063:1085 synaptic iGluR function 1063:1085 These results lead to a model where protein and small-molecule ligands synergistically control synaptic iGluR function. 27418511 1 39 theme family 126:131 arg1 members 133:139 Ionotropic glutamate receptor (iGluR) family members 88:139 Ionotropic glutamate receptor (iGluR) family members 88:139 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 5 40 theme PF-PC 909:913 arg1 synapses 916:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 1 41 theme excitatory 231:240 arg1 synapses 242:249 excitatory synapses 231:249 excitatory synapses 231:249 Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. 27418511 3 42 theme iGluR 531:535 arg1 δ2 537:538 postsynaptic iGluR δ2 518:538 postsynaptic iGluR δ2 (GluD2) 518:546 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 3 42 theme iGluR 531:535 arg1 GluD2 541:545 GluD2 541:545 GluD2 541:545 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 3 43 theme postsynaptic 518:529 arg1 δ2 537:538 postsynaptic iGluR δ2 518:538 postsynaptic iGluR δ2 (GluD2) 518:546 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 3 43 theme postsynaptic 518:529 arg1 GluD2 541:545 GluD2 541:545 GluD2 541:545 Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. 27418511 0 44 theme organizer 67:75 arg1 complexes 77:85 synaptic organizer complexes 58:85 synaptic organizer complexes 58:85 Structural basis for integration of GluD receptors within synaptic organizer complexes. 27418511 5 45 theme synapses 916:923 arg1 depression 854:863 long-term depression 844:863 long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice 844:965 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 2 46 theme iGluR 405:409 arg1 signaling 411:419 physiological iGluR signaling 391:419 physiological iGluR signaling 391:419 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 27418511 5 47 theme cell 903:906 arg1 synapses 916:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 cerebellar parallel fiber-Purkinje cell (PF-PC) synapses 868:923 This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. 27418511 0 48 theme synaptic 58:65 arg1 complexes 77:85 synaptic organizer complexes 58:85 synaptic organizer complexes 58:85 Structural basis for integration of GluD receptors within synaptic organizer complexes. 27418511 2 49 theme physiological 391:403 arg1 signaling 411:419 physiological iGluR signaling 391:419 physiological iGluR signaling 391:419 However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. 29692406 8 0 theme protein 1046:1052 arg1 fold 1054:1057 the protein fold 1042:1057 the protein fold 1042:1057 Structural mapping of disease mutations reveals that most would destabilize the protein fold. 29692406 6 1 theme distinct 697:704 arg1 mechanisms 716:725 distinct catalytic mechanisms 697:725 distinct catalytic mechanisms for substrate hydrolysis versus autocleavage 697:770 Substrate modeling suggests distinct catalytic mechanisms for substrate hydrolysis versus autocleavage. 29692406 5 2 theme hydrophobic 621:631 arg1 channel 633:639 a hydrophobic channel 619:639 a hydrophobic channel leading to the active site 619:666 Autocleavage triggers a conformational change exposing a hydrophobic channel leading to the active site. 29692406 8 3 theme disease 988:994 arg1 mutations 996:1004 disease mutations 988:1004 disease mutations 988:1004 Structural mapping of disease mutations reveals that most would destabilize the protein fold. 29692406 7 4 theme attachment 870:879 arg1 site 853:856 a site 851:856 a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D 851:963 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 7 4 theme attachment 870:879 arg1 attachment 870:879 membrane attachment 861:879 membrane attachment 861:879 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 7 4 theme attachment 870:879 arg1 surface 787:793 A hydrophobic surface 773:793 A hydrophobic surface surrounding the substrate binding channel 773:835 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 2 5 theme myoclonic 311:319 arg1 epilepsy 321:328 progressive myoclonic epilepsy 299:328 progressive myoclonic epilepsy 299:328 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 8 6 theme Structural 966:975 arg1 mapping 977:983 Structural mapping 966:983 Structural mapping of disease mutations 966:1004 Structural mapping of disease mutations reveals that most would destabilize the protein fold. 29692406 1 7 theme Acid 56:59 arg1 ceramidase 61:70 Acid ceramidase 56:70 Acid ceramidase (aCDase, ASAH1) 56:86 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 2 8 with atrophy 286:292 arg1 epilepsy 321:328 progressive myoclonic epilepsy 299:328 progressive myoclonic epilepsy 299:328 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 8 9 theme mutations 996:1004 arg1 mapping 977:983 Structural mapping 966:983 Structural mapping of disease mutations 966:1004 Structural mapping of disease mutations reveals that most would destabilize the protein fold. 29692406 2 10 theme Farber 254:259 arg1 disease 261:267 Farber disease 254:267 Farber disease 254:267 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 2 10 theme Farber 254:259 arg1 atrophy 286:292 spinal muscular atrophy 270:292 spinal muscular atrophy with progressive myoclonic epilepsy 270:328 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 7 11 theme membrane 861:868 arg1 attachment 870:879 membrane attachment 861:879 membrane attachment 861:879 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 5 12 theme conformational 588:601 arg1 change 603:608 a conformational change 586:608 a conformational change exposing a hydrophobic channel leading to the active site 586:666 Autocleavage triggers a conformational change exposing a hydrophobic channel leading to the active site. 29692406 6 13 theme Substrate 669:677 arg1 modeling 679:686 Substrate modeling 669:686 Substrate modeling 669:686 Substrate modeling suggests distinct catalytic mechanisms for substrate hydrolysis versus autocleavage. 29692406 7 14 theme substrate 811:819 arg1 channel 829:835 the substrate binding channel 807:835 the substrate binding channel 807:835 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 0 15 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for the activation of acid ceramidase 0:53 Structural basis for the activation of acid ceramidase. 29692406 4 16 theme catalytic 509:517 arg1 center 519:524 the catalytic center 505:524 the catalytic center 505:524 In the proenzyme, the catalytic center is buried and protected from solvent. 29692406 9 17 theme aCDase 1109:1114 arg1 inhibitors 1116:1125 aCDase inhibitors 1109:1125 aCDase inhibitors 1109:1125 These results will inform the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics. 29692406 1 18 theme sphingolipids 165:177 arg1 sphingosine 132:142 sphingosine 132:142 sphingosine 132:142 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 1 18 theme sphingolipids 165:177 arg1 backbone 149:156 the backbone 145:156 the backbone of all sphingolipids 145:177 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 7 19 theme binding 821:827 arg1 channel 829:835 the substrate binding channel 807:835 the substrate binding channel 807:835 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 7 20 theme accessory 936:944 arg1 protein 946:952 the accessory protein 932:952 the accessory protein 932:952 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 7 20 theme accessory 936:944 arg1 saposin-D 955:963 saposin-D 955:963 saposin-D 955:963 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 2 21 theme muscular 277:284 arg1 disease 261:267 Farber disease 254:267 Farber disease 254:267 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 2 21 theme muscular 277:284 arg1 atrophy 286:292 spinal muscular atrophy 270:292 spinal muscular atrophy with progressive myoclonic epilepsy 270:328 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 9 22 theme inhibitors 1116:1125 arg1 design 1099:1104 the rational design 1086:1104 the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics 1086:1173 These results will inform the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics. 29692406 3 23 dep both 449:452 arg1 proenzyme 454:462 proenzyme 454:462 proenzyme 454:462 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 2 24 theme spinal 270:275 arg1 disease 261:267 Farber disease 254:267 Farber disease 254:267 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 2 24 theme spinal 270:275 arg1 atrophy 286:292 spinal muscular atrophy 270:292 spinal muscular atrophy with progressive myoclonic epilepsy 270:328 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 3 25 theme mammalian 428:436 arg1 aCDases 438:444 mammalian aCDases 428:444 mammalian aCDases 428:444 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 9 26 theme rational 1090:1097 arg1 design 1099:1104 the rational design 1086:1104 the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics 1086:1173 These results will inform the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics. 29692406 2 27 theme Abnormal 217:224 arg1 function 226:233 Abnormal function 217:233 Abnormal function of aCDase 217:243 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 9 28 theme recombinant 1131:1141 arg1 aCDase 1143:1148 recombinant aCDase 1131:1148 recombinant aCDase 1131:1148 These results will inform the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics. 29692406 3 29 theme crystal 406:412 arg1 structures 414:423 crystal structures 406:423 crystal structures of mammalian aCDases in both proenzyme and autocleaved forms 406:484 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 7 30 theme hydrophobic 775:785 arg1 site 853:856 a site 851:856 a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D 851:963 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 7 30 theme hydrophobic 775:785 arg1 attachment 870:879 membrane attachment 861:879 membrane attachment 861:879 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 7 30 theme hydrophobic 775:785 arg1 surface 787:793 A hydrophobic surface 773:793 A hydrophobic surface surrounding the substrate binding channel 773:835 A hydrophobic surface surrounding the substrate binding channel appears to be a site of membrane attachment where the enzyme accepts substrates facilitated by the accessory protein, saposin-D. 29692406 9 31 theme aCDase 1143:1148 arg1 design 1099:1104 the rational design 1086:1104 the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics 1086:1173 These results will inform the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics. 29692406 1 32 theme many 192:195 arg1 processes 206:214 many cellular processes 192:214 many cellular processes 192:214 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 0 33 theme ceramidase 44:53 arg1 activation 25:34 the activation 21:34 the activation of acid ceramidase 21:53 Structural basis for the activation of acid ceramidase. 29692406 3 34 from structures 414:423 arg1 forms 480:484 both proenzyme and autocleaved forms 449:484 both proenzyme and autocleaved forms 449:484 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 1 35 dep ceramidase 61:70 arg1 ASAH1 81:85 ASAH1 81:85 ASAH1 81:85 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 1 35 dep ceramidase 61:70 arg1 aCDase 73:78 aCDase 73:78 aCDase 73:78 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 0 36 theme acid 39:42 arg1 ceramidase 44:53 acid ceramidase 39:53 acid ceramidase 39:53 Structural basis for the activation of acid ceramidase. 29692406 9 37 theme disease 1154:1160 arg1 therapeutics 1162:1173 disease therapeutics 1154:1173 disease therapeutics 1154:1173 These results will inform the rational design of aCDase inhibitors and recombinant aCDase for disease therapeutics. 29692406 1 38 theme lysosomal 99:107 arg1 ceramide 118:125 lysosomal membrane ceramide 99:125 lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, 99:178 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 5 39 theme active 656:661 arg1 site 663:666 the active site 652:666 the active site 652:666 Autocleavage triggers a conformational change exposing a hydrophobic channel leading to the active site. 29692406 6 40 theme substrate 731:739 arg1 hydrolysis 741:750 substrate hydrolysis 731:750 substrate hydrolysis 731:750 Substrate modeling suggests distinct catalytic mechanisms for substrate hydrolysis versus autocleavage. 29692406 1 41 theme membrane 109:116 arg1 ceramide 118:125 lysosomal membrane ceramide 99:125 lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, 99:178 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 3 42 dep forms 480:484 arg1 both 449:452 both 449:452 both 449:452 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 3 42 dep forms 480:484 arg1 autocleaved 468:478 autocleaved 468:478 autocleaved 468:478 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 1 43 theme cellular 197:204 arg1 processes 206:214 many cellular processes 192:214 many cellular processes 192:214 Acid ceramidase (aCDase, ASAH1) hydrolyzes lysosomal membrane ceramide into sphingosine, the backbone of all sphingolipids, to regulate many cellular processes. 29692406 2 44 theme aCDase 238:243 arg1 function 226:233 Abnormal function 217:233 Abnormal function of aCDase 217:243 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 2 45 theme progressive 299:309 arg1 epilepsy 321:328 progressive myoclonic epilepsy 299:328 progressive myoclonic epilepsy 299:328 Abnormal function of aCDase leads to Farber disease, spinal muscular atrophy with progressive myoclonic epilepsy, and is associated with Alzheimer's, diabetes, and cancer. 29692406 3 46 theme aCDases 438:444 arg1 structures 414:423 crystal structures 406:423 crystal structures of mammalian aCDases in both proenzyme and autocleaved forms 406:484 Here, we present crystal structures of mammalian aCDases in both proenzyme and autocleaved forms. 29692406 6 47 theme catalytic 706:714 arg1 mechanisms 716:725 distinct catalytic mechanisms 697:725 distinct catalytic mechanisms for substrate hydrolysis versus autocleavage 697:770 Substrate modeling suggests distinct catalytic mechanisms for substrate hydrolysis versus autocleavage. 25922362 6 0 theme spectrometry 1302:1313 arg1 analysis 1315:1322 mass spectrometry analysis 1297:1322 mass spectrometry analysis 1297:1322 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 2 1 theme recent 571:576 arg1 therapies 604:612 most recent monoclonal antibody-based therapies 566:612 most recent monoclonal antibody-based therapies against CRC 566:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 2 theme membrane-associated 342:360 arg1 NEU3 372:375 The plasma membrane-associated sialidase NEU3 331:375 The plasma membrane-associated sialidase NEU3 331:375 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 4 3 theme CCD841 980:985 arg1 line 1010:1013 the CCD841 normal intestinal cell line 976:1013 the CCD841 normal intestinal cell line 976:1013 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 3 4 theme cell 788:791 arg1 lines 793:797 commercial colon cell lines 771:797 commercial colon cell lines 771:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 6 5 theme mass 1297:1300 arg1 analysis 1315:1322 mass spectrometry analysis 1297:1322 mass spectrometry analysis 1297:1322 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 3 6 theme sialidase 659:667 arg1 role 646:649 the role 642:649 the role of NEU3 sialidase on EGFR deregulation in CRC 642:695 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 2 7 theme most 566:569 arg1 therapies 604:612 most recent monoclonal antibody-based therapies 566:612 most recent monoclonal antibody-based therapies against CRC 566:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 8 theme plasma 335:340 arg1 NEU3 372:375 The plasma membrane-associated sialidase NEU3 331:375 The plasma membrane-associated sialidase NEU3 331:375 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 4 9 theme cell 1005:1008 arg1 line 1010:1013 the CCD841 normal intestinal cell line 976:1013 the CCD841 normal intestinal cell line 976:1013 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 3 10 theme lines 793:797 arg1 library 760:766 a library 758:766 a library of commercial colon cell lines 758:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 0 11 theme sialylation 89:99 arg1 levels 101:106 its sialylation levels 85:106 its sialylation levels 85:106 NEU3 activity enhances EGFR activation without affecting EGFR expression and acts on its sialylation levels. 25922362 4 12 theme intestinal 994:1003 arg1 line 1010:1013 the CCD841 normal intestinal cell line 976:1013 the CCD841 normal intestinal cell line 976:1013 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 3 13 theme commercial 771:780 arg1 lines 793:797 commercial colon cell lines 771:797 commercial colon cell lines 771:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 2 14 theme epidermal 501:509 arg1 target 556:561 the molecular target 542:561 the molecular target of most recent monoclonal antibody-based therapies against CRC 542:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 14 theme epidermal 501:509 arg1 EGFR 535:538 EGFR 535:538 EGFR 535:538 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 14 theme epidermal 501:509 arg1 receptor 525:532 epidermal growth factor receptor 501:532 epidermal growth factor receptor (EGFR) 501:539 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 5 15 theme inactive 1098:1105 arg1 form 1114:1117 the inactive mutant form 1094:1117 the inactive mutant form of NEU3 1094:1125 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 3 16 theme colon 782:786 arg1 lines 793:797 commercial colon cell lines 771:797 commercial colon cell lines 771:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 6 17 theme western 1279:1285 arg1 blots 1287:1291 western blots 1279:1291 western blots 1279:1291 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 1 18 gly sialylation 184:194 arg1 cancer 218:223 cancer 218:223 cancer 218:223 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 2 19 theme therapies 604:612 arg1 target 556:561 the molecular target 542:561 the molecular target of most recent monoclonal antibody-based therapies against CRC 542:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 19 theme therapies 604:612 arg1 receptor 525:532 epidermal growth factor receptor 501:532 epidermal growth factor receptor (EGFR) 501:539 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 4 20 theme SW620 960:964 arg1 cells 966:970 SW620 cells 960:970 SW620 cells 960:970 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 3 21 theme NEU3 725:728 arg1 levels 746:751 NEU3 gene expression levels 725:751 NEU3 gene expression levels from a library of commercial colon cell lines 725:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 4 22 theme cells 966:970 arg1 exception 947:955 the exception 943:955 the exception of SW620 cells and the CCD841 normal intestinal cell line 943:1013 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 2 23 theme antibody-based 589:602 arg1 therapies 604:612 most recent monoclonal antibody-based therapies 566:612 most recent monoclonal antibody-based therapies against CRC 566:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 4 24 theme cell 926:929 arg1 lines 931:935 all cell lines 922:935 all cell lines 922:935 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 3 25 theme cell 862:865 arg1 lines 867:871 these cell lines 856:871 these cell lines 856:871 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 6 26 theme inactive 1463:1470 arg1 NEU3 1472:1475 inactive NEU3 1463:1475 inactive NEU3 1463:1475 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 0 27 theme NEU3 0:3 arg1 activity 5:12 NEU3 activity 0:12 NEU3 activity 0:12 NEU3 activity enhances EGFR activation without affecting EGFR expression and acts on its sialylation levels. 25922362 5 28 theme mutant 1107:1112 arg1 form 1114:1117 the inactive mutant form 1094:1117 the inactive mutant form of NEU3 1094:1125 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 2 29 theme monoclonal 578:587 arg1 therapies 604:612 most recent monoclonal antibody-based therapies 566:612 most recent monoclonal antibody-based therapies against CRC 566:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 6 30 from cells 1432:1436 arg1 receptor 1413:1420 the receptor 1409:1420 the receptor from mock cells and cells overexpressing inactive NEU3 1409:1475 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 5 31 theme EGFR 1228:1231 arg1 mRNA 1233:1236 EGFR mRNA 1228:1236 EGFR mRNA 1228:1236 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 0 32 theme EGFR 23:26 arg1 activation 28:37 EGFR activation 23:37 EGFR activation 23:37 NEU3 activity enhances EGFR activation without affecting EGFR expression and acts on its sialylation levels. 25922362 5 33 theme form 1114:1117 arg1 overexpression 1052:1065 overexpression 1052:1065 overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR 1052:1133 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 7 34 theme EGFR 1644:1647 arg1 activation 1649:1658 EGFR activation 1644:1658 EGFR activation through its desialylation 1644:1684 On the whole, our data demonstrate that, besides the already reported indirect EGFR activation through GM3, sialidase NEU3 could also play a role on EGFR activation through its desialylation. 25922362 6 35 from cells 1442:1446 arg1 receptor 1413:1420 the receptor 1409:1420 the receptor from mock cells and cells overexpressing inactive NEU3 1409:1475 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 7 36 theme EGFR 1574:1577 arg1 activation 1579:1588 the already reported indirect EGFR activation 1544:1588 the already reported indirect EGFR activation through GM3 1544:1600 On the whole, our data demonstrate that, besides the already reported indirect EGFR activation through GM3, sialidase NEU3 could also play a role on EGFR activation through its desialylation. 25922362 1 37 theme sialylation 184:194 arg1 role 176:179 the role 172:179 the role of sialylation in the progression of cancer and, in particular 172:242 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 5 38 theme NEU3 1122:1125 arg1 form 1114:1117 the inactive mutant form 1094:1117 the inactive mutant form of NEU3 1094:1125 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 5 39 from overexpression 1052:1065 arg1 EGFR 1130:1133 EGFR 1130:1133 EGFR 1130:1133 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 5 40 theme NEU3 1176:1179 arg1 form 1168:1171 the active form 1157:1171 the active form of NEU3 1157:1179 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 5 41 theme protein 1241:1247 arg1 expression 1249:1258 protein expression 1241:1258 protein expression 1241:1258 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 6 42 theme mock 1427:1430 arg1 cells 1432:1436 mock cells 1427:1436 mock cells 1427:1436 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 3 43 theme expression 735:744 arg1 levels 746:751 NEU3 gene expression levels 725:751 NEU3 gene expression levels from a library of commercial colon cell lines 725:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 2 44 theme factor 518:523 arg1 target 556:561 the molecular target 542:561 the molecular target of most recent monoclonal antibody-based therapies against CRC 542:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 44 theme factor 518:523 arg1 EGFR 535:538 EGFR 535:538 EGFR 535:538 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 44 theme factor 518:523 arg1 receptor 525:532 epidermal growth factor receptor 501:532 epidermal growth factor receptor (EGFR) 501:539 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 6 45 gly desialylated 1481:1492 arg1 EGFR 1339:1342 EGFR 1339:1342 EGFR immunoprecipitated from cells overexpressing active NEU3 1339:1399 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 2 46 theme colorectal 401:410 arg1 cancer 412:417 colorectal cancer 401:417 colorectal cancer (CRC) 401:423 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 46 theme colorectal 401:410 arg1 CRC 420:422 CRC 420:422 CRC 420:422 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 0 47 theme EGFR 57:60 arg1 expression 62:71 EGFR expression 57:71 EGFR expression 57:71 NEU3 activity enhances EGFR activation without affecting EGFR expression and acts on its sialylation levels. 25922362 2 48 theme growth 511:516 arg1 target 556:561 the molecular target 542:561 the molecular target of most recent monoclonal antibody-based therapies against CRC 542:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 48 theme growth 511:516 arg1 EGFR 535:538 EGFR 535:538 EGFR 535:538 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 48 theme growth 511:516 arg1 receptor 525:532 epidermal growth factor receptor 501:532 epidermal growth factor receptor (EGFR) 501:539 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 49 theme molecular 546:554 arg1 target 556:561 the molecular target 542:561 the molecular target of most recent monoclonal antibody-based therapies against CRC 542:624 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 49 theme molecular 546:554 arg1 receptor 525:532 epidermal growth factor receptor 501:532 epidermal growth factor receptor (EGFR) 501:539 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 7 50 theme indirect 1565:1572 arg1 activation 1579:1588 the already reported indirect EGFR activation 1544:1588 the already reported indirect EGFR activation through GM3 1544:1600 On the whole, our data demonstrate that, besides the already reported indirect EGFR activation through GM3, sialidase NEU3 could also play a role on EGFR activation through its desialylation. 25922362 1 51 theme Several 109:115 arg1 studies 117:123 Several studies 109:123 Several studies performed over the last decade 109:154 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 3 52 from deregulation 677:688 arg1 CRC 693:695 CRC 693:695 CRC 693:695 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 1 53 theme sialidases 288:297 arg1 transformation 315:328 tumorigenic transformation 303:328 tumorigenic transformation 303:328 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 1 53 theme sialidases 288:297 arg1 deregulation 272:283 deregulation 272:283 deregulation of sialidases 272:297 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 3 54 theme EGFR 672:675 arg1 deregulation 677:688 EGFR deregulation 672:688 EGFR deregulation in CRC 672:695 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 7 55 theme sialidase 1603:1611 arg1 NEU3 1613:1616 sialidase NEU3 1603:1616 sialidase NEU3 1603:1616 On the whole, our data demonstrate that, besides the already reported indirect EGFR activation through GM3, sialidase NEU3 could also play a role on EGFR activation through its desialylation. 25922362 5 56 theme wild-type 1081:1089 arg1 overexpression 1052:1065 overexpression 1052:1065 overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR 1052:1133 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 6 57 theme active 1389:1394 arg1 NEU3 1396:1399 active NEU3 1389:1399 active NEU3 1389:1399 Moreover, through western blots and mass spectrometry analysis, we found that EGFR immunoprecipitated from cells overexpressing active NEU3, unlike the receptor from mock cells and cells overexpressing inactive NEU3, is desialylated. 25922362 5 58 theme receptor 1190:1197 arg1 activation 1199:1208 receptor activation 1190:1208 receptor activation 1190:1208 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 3 59 theme gene 730:733 arg1 levels 746:751 NEU3 gene expression levels 725:751 NEU3 gene expression levels from a library of commercial colon cell lines 725:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 3 60 from library 760:766 arg1 levels 746:751 NEU3 gene expression levels 725:751 NEU3 gene expression levels from a library of commercial colon cell lines 725:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 3 61 theme NEU3 654:657 arg1 sialidase 659:667 NEU3 sialidase 654:667 NEU3 sialidase 654:667 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 3 62 theme NEU3 819:822 arg1 transcription 824:836 NEU3 transcription 819:836 NEU3 transcription 819:836 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 5 63 from EGFR 1130:1133 arg1 overexpression 1052:1065 overexpression 1052:1065 overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR 1052:1133 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 25922362 7 64 theme reported 1556:1563 arg1 activation 1579:1588 the already reported indirect EGFR activation 1544:1588 the already reported indirect EGFR activation through GM3 1544:1600 On the whole, our data demonstrate that, besides the already reported indirect EGFR activation through GM3, sialidase NEU3 could also play a role on EGFR activation through its desialylation. 25922362 2 65 theme sialidase 362:370 arg1 NEU3 372:375 The plasma membrane-associated sialidase NEU3 331:375 The plasma membrane-associated sialidase NEU3 331:375 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 2 66 theme HeLa 485:488 arg1 cells 490:494 HeLa cells 485:494 HeLa cells 485:494 The plasma membrane-associated sialidase NEU3 is often deregulated in colorectal cancer (CRC), and it was shown that this enzyme co-immunoprecipitates in HeLa cells with epidermal growth factor receptor (EGFR), the molecular target of most recent monoclonal antibody-based therapies against CRC. 25922362 3 67 from data 717:720 arg1 levels 746:751 NEU3 gene expression levels 725:751 NEU3 gene expression levels from a library of commercial colon cell lines 725:797 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 1 68 from role 176:179 arg1 progression 203:213 the progression 199:213 the progression of cancer 199:223 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 1 68 from role 176:179 arg1 particular 233:242 particular 233:242 particular 233:242 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 4 69 theme line 1010:1013 arg1 exception 947:955 the exception 943:955 the exception of SW620 cells and the CCD841 normal intestinal cell line 943:1013 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 1 70 theme cancer 218:223 arg1 progression 203:213 the progression 199:213 the progression of cancer 199:223 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 3 71 from role 646:649 arg1 deregulation 677:688 EGFR deregulation 672:688 EGFR deregulation in CRC 672:695 To investigate the role of NEU3 sialidase on EGFR deregulation in CRC, we first collected data on NEU3 gene expression levels from a library of commercial colon cell lines, demonstrating that NEU3 transcription is upregulated in these cell lines. 25922362 1 72 theme tumorigenic 303:313 arg1 transformation 315:328 tumorigenic transformation 303:328 tumorigenic transformation 303:328 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 1 73 theme last 144:147 arg1 decade 149:154 the last decade 140:154 the last decade 140:154 Several studies performed over the last decade have focused on the role of sialylation in the progression of cancer and, in particular, on the association between deregulation of sialidases and tumorigenic transformation. 25922362 4 74 theme normal 987:992 arg1 line 1010:1013 the CCD841 normal intestinal cell line 976:1013 the CCD841 normal intestinal cell line 976:1013 We also found EGFR to be hyperphosphorylated in all cell lines, with the exception of SW620 cells and the CCD841 normal intestinal cell line. 25922362 5 75 theme active 1161:1166 arg1 form 1168:1171 the active form 1157:1171 the active form of NEU3 1157:1179 By comparing the effects induced by overexpression of either the wild-type or the inactive mutant form of NEU3 on EGFR, we demonstrated that the active form of NEU3 enhanced receptor activation without affecting EGFR mRNA or protein expression. 33837739 7 0 theme variants 1347:1354 arg1 mistrafficking 1311:1324 intracellular mistrafficking 1297:1324 intracellular mistrafficking of the AE-associated variants 1297:1354 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 8 1 theme pathogenic 1437:1446 arg1 mechanism 1448:1456 a molecular pathogenic mechanism 1425:1456 a molecular pathogenic mechanism for AE 1425:1463 This work provides a molecular pathogenic mechanism for AE. 33837739 1 2 theme ZIP 171:173 arg1 family 188:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 3 3 attach present 570:576 arg2 domain 533:538 the extracellular domain 515:538 the extracellular domain (ECD) 515:544 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 3 attach present 570:576 arg2 present 570:576 present 570:576 present 570:576 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 3 attach present 570:576 arg2 ECD 541:543 ECD 541:543 ECD 541:543 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 3 attach present 570:576 arg1 fraction 583:590 a fraction 581:590 a fraction of mammalian ZIPs 581:608 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 3 attach present 570:576 arg2 hZIP4 549:553 hZIP4 549:553 hZIP4 549:553 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 7 4 theme Pteropus 1156:1163 arg1 Alecto 1165:1170 Pteropus Alecto 1156:1170 Pteropus Alecto 1156:1170 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 4 theme Pteropus 1156:1163 arg1 homolog 1181:1187 a close homolog 1173:1187 a close homolog of hZIP4 1173:1196 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 3 5 theme ZIPs 605:608 arg1 fraction 583:590 a fraction 581:590 a fraction of mammalian ZIPs 581:608 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 6 theme extracellular 519:531 arg1 hZIP4 549:553 hZIP4 549:553 hZIP4 549:553 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 6 theme extracellular 519:531 arg1 ECD 541:543 ECD 541:543 ECD 541:543 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 6 theme extracellular 519:531 arg1 present 570:576 present 570:576 present 570:576 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 6 theme extracellular 519:531 arg1 domain 533:538 the extracellular domain 515:538 the extracellular domain (ECD) 515:544 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 7 7 theme zinc 1384:1387 arg1 activity 1396:1403 zinc uptake activity 1384:1403 zinc uptake activity 1384:1403 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 6 8 theme cell 1005:1008 arg1 surface 1010:1016 the cell surface 1001:1016 the cell surface 1001:1016 Although the variants were able to be expressed in HEK293T cells, they failed to traffic to the cell surface and were largely retained in the ER with immature glycosylation. 33837739 7 9 theme activity 1396:1403 arg1 loss 1376:1379 such a total loss 1363:1379 such a total loss of zinc uptake activity 1363:1403 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 10 theme hZIP4 1192:1196 arg1 Alecto 1165:1170 Pteropus Alecto 1156:1170 Pteropus Alecto 1156:1170 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 10 theme hZIP4 1192:1196 arg1 homolog 1181:1187 a close homolog 1173:1187 a close homolog of hZIP4 1173:1196 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 11 theme uptake 1389:1394 arg1 activity 1396:1403 zinc uptake activity 1384:1403 zinc uptake activity 1384:1403 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 3 12 from fraction 583:590 arg1 present 570:576 present 570:576 present 570:576 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 12 from fraction 583:590 arg1 hZIP4 549:553 hZIP4 549:553 hZIP4 549:553 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 12 from fraction 583:590 arg1 domain 533:538 the extracellular domain 515:538 the extracellular domain (ECD) 515:544 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 4 13 theme AE-causing 621:630 arg1 mutations 636:644 these AE-causing ECD mutations 615:644 these AE-causing ECD mutations 615:644 How these AE-causing ECD mutations lead to ZIP4 malfunction has not be fully clarified. 33837739 7 14 theme ZIP4 1146:1149 arg1 ECD 1139:1141 the ECD 1135:1141 the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4 1135:1196 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 1 15 theme representative 119:132 arg1 ZIP4 109:112 ZIP4 109:112 ZIP4 109:112 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 1 15 theme representative 119:132 arg1 member 134:139 a representative member 117:139 a representative member of the Zrt-/Irt-like protein (ZIP) transporter family 117:193 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 0 16 attach linked 27:32 arg1 enteropathica 52:64 acrodermatitis enteropathica 37:64 acrodermatitis enteropathica 37:64 Zinc transporter mutations linked to acrodermatitis enteropathica disrupt function and cause mistrafficking. 33837739 0 16 attach linked 27:32 arg2 mutations 17:25 Zinc transporter mutations 0:25 Zinc transporter mutations linked to acrodermatitis enteropathica 0:64 Zinc transporter mutations linked to acrodermatitis enteropathica disrupt function and cause mistrafficking. 33837739 5 17 theme zinc 852:855 arg1 activity 867:874 completely abolished zinc transport activity 831:874 completely abolished zinc transport activity 831:874 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 6 18 theme HEK293T 960:966 arg1 cells 968:972 HEK293T cells 960:972 HEK293T cells 960:972 Although the variants were able to be expressed in HEK293T cells, they failed to traffic to the cell surface and were largely retained in the ER with immature glycosylation. 33837739 5 19 theme cell-based 881:890 arg1 assay 902:906 a cell-based transport assay 879:906 a cell-based transport assay 879:906 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 5 20 theme confirmed 740:748 arg1 mutations 770:778 all seven confirmed AE-causing missense mutations 730:778 all seven confirmed AE-causing missense mutations 730:778 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 7 21 dep exhibited 1212:1220 arg1 accounts 1284:1291 accounts 1284:1291 accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity 1284:1403 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 1 22 theme transporter 176:186 arg1 family 188:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 0 23 theme transporter 5:15 arg1 mutations 17:25 Zinc transporter mutations 0:25 Zinc transporter mutations linked to acrodermatitis enteropathica 0:64 Zinc transporter mutations linked to acrodermatitis enteropathica disrupt function and cause mistrafficking. 33837739 1 24 theme family 188:193 arg1 ZIP4 109:112 ZIP4 109:112 ZIP4 109:112 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 1 24 theme family 188:193 arg1 member 134:139 a representative member 117:139 a representative member of the Zrt-/Irt-like protein (ZIP) transporter family 117:193 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 0 25 theme Zinc 0:3 arg1 mutations 17:25 Zinc transporter mutations 0:25 Zinc transporter mutations linked to acrodermatitis enteropathica 0:64 Zinc transporter mutations linked to acrodermatitis enteropathica disrupt function and cause mistrafficking. 33837739 8 26 theme molecular 1427:1435 arg1 mechanism 1448:1456 a molecular pathogenic mechanism 1425:1456 a molecular pathogenic mechanism for AE 1425:1463 This work provides a molecular pathogenic mechanism for AE. 33837739 5 27 theme abolished 842:850 arg1 activity 867:874 completely abolished zinc transport activity 831:874 completely abolished zinc transport activity 831:874 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 5 28 theme transport 857:865 arg1 activity 867:874 completely abolished zinc transport activity 831:874 completely abolished zinc transport activity 831:874 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 3 29 theme transport 482:490 arg1 machinery 492:500 the conserved transmembrane zinc transport machinery 449:500 the conserved transmembrane zinc transport machinery 449:500 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 2 30 theme zinc 306:309 arg1 absorption 311:320 zinc absorption 306:320 zinc absorption 306:320 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 3 31 from present 570:576 arg1 fraction 583:590 a fraction 581:590 a fraction of mammalian ZIPs 581:608 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 32 theme zinc 477:480 arg1 machinery 492:500 the conserved transmembrane zinc transport machinery 449:500 the conserved transmembrane zinc transport machinery 449:500 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 7 33 theme total 1370:1374 arg1 loss 1376:1379 such a total loss 1363:1379 such a total loss of zinc uptake activity 1363:1403 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 2 34 theme recessive 360:368 arg1 disorder 370:377 a life-threatening autosomal recessive disorder 331:377 a life-threatening autosomal recessive disorder 331:377 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 2 34 theme recessive 360:368 arg1 enteropathica 395:407 acrodermatitis enteropathica 380:407 acrodermatitis enteropathica (AE) 380:412 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 1 35 theme Zrt-/Irt-like 148:160 arg1 family 188:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 0 36 theme acrodermatitis 37:50 arg1 enteropathica 52:64 acrodermatitis enteropathica 37:64 acrodermatitis enteropathica 37:64 Zinc transporter mutations linked to acrodermatitis enteropathica disrupt function and cause mistrafficking. 33837739 3 37 located present 570:576 arg2 domain 533:538 the extracellular domain 515:538 the extracellular domain (ECD) 515:544 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 37 located present 570:576 arg2 present 570:576 present 570:576 present 570:576 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 37 located present 570:576 arg2 ECD 541:543 ECD 541:543 ECD 541:543 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 37 located present 570:576 arg1 fraction 583:590 a fraction 581:590 a fraction of mammalian ZIPs 581:608 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 37 located present 570:576 arg2 hZIP4 549:553 hZIP4 549:553 hZIP4 549:553 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 2 38 theme Loss-of-function 238:253 arg1 mutations 255:263 Loss-of-function mutations 238:263 Loss-of-function mutations of human ZIP4 (hZIP4) 238:285 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 1 39 theme zinc 215:218 arg1 uptake 220:225 zinc uptake 215:225 zinc uptake from diet 215:235 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 5 40 theme AE-causing 750:759 arg1 mutations 770:778 all seven confirmed AE-causing missense mutations 730:778 all seven confirmed AE-causing missense mutations 730:778 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 3 41 theme transmembrane 463:475 arg1 machinery 492:500 the conserved transmembrane zinc transport machinery 449:500 the conserved transmembrane zinc transport machinery 449:500 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 5 42 theme missense 761:768 arg1 mutations 770:778 all seven confirmed AE-causing missense mutations 730:778 all seven confirmed AE-causing missense mutations 730:778 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 7 43 from Alecto 1165:1170 arg1 ECD 1139:1141 the ECD 1135:1141 the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4 1135:1196 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 6 44 theme immature 1059:1066 arg1 glycosylation 1068:1080 immature glycosylation 1059:1080 immature glycosylation 1059:1080 Although the variants were able to be expressed in HEK293T cells, they failed to traffic to the cell surface and were largely retained in the ER with immature glycosylation. 33837739 2 45 theme autosomal 350:358 arg1 disorder 370:377 a life-threatening autosomal recessive disorder 331:377 a life-threatening autosomal recessive disorder 331:377 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 2 45 theme autosomal 350:358 arg1 enteropathica 395:407 acrodermatitis enteropathica 380:407 acrodermatitis enteropathica (AE) 380:412 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 7 46 theme AE-associated 1333:1345 arg1 variants 1347:1354 the AE-associated variants 1329:1354 the AE-associated variants 1329:1354 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 47 theme structural 1222:1231 arg1 defects 1233:1239 structural defects 1222:1239 structural defects 1222:1239 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 2 48 theme life-threatening 333:348 arg1 disorder 370:377 a life-threatening autosomal recessive disorder 331:377 a life-threatening autosomal recessive disorder 331:377 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 2 48 theme life-threatening 333:348 arg1 enteropathica 395:407 acrodermatitis enteropathica 380:407 acrodermatitis enteropathica (AE) 380:412 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 3 49 theme mammalian 595:603 arg1 ZIPs 605:608 mammalian ZIPs 595:608 mammalian ZIPs 595:608 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 2 50 theme ZIP4 274:277 arg1 mutations 255:263 Loss-of-function mutations 238:263 Loss-of-function mutations of human ZIP4 (hZIP4) 238:285 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 3 51 theme hZIP4 549:553 arg1 hZIP4 549:553 hZIP4 549:553 hZIP4 549:553 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 51 theme hZIP4 549:553 arg1 ECD 541:543 ECD 541:543 ECD 541:543 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 51 theme hZIP4 549:553 arg1 present 570:576 present 570:576 present 570:576 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 3 51 theme hZIP4 549:553 arg1 domain 533:538 the extracellular domain 515:538 the extracellular domain (ECD) 515:544 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 1 52 from diet 232:235 arg1 uptake 220:225 zinc uptake 215:225 zinc uptake from diet 215:235 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 33837739 5 53 theme transport 892:900 arg1 assay 902:906 a cell-based transport assay 879:906 a cell-based transport assay 879:906 In this work, we characterized all seven confirmed AE-causing missense mutations in hZIP4-ECD and found that the variants exhibited completely abolished zinc transport activity in a cell-based transport assay. 33837739 2 54 theme human 268:272 arg1 hZIP4 280:284 hZIP4 280:284 hZIP4 280:284 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 2 54 theme human 268:272 arg1 ZIP4 274:277 human ZIP4 268:277 human ZIP4 (hZIP4) 268:285 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 4 55 theme ZIP4 654:657 arg1 malfunction 659:669 ZIP4 malfunction 654:669 ZIP4 malfunction 654:669 How these AE-causing ECD mutations lead to ZIP4 malfunction has not be fully clarified. 33837739 7 56 theme close 1175:1179 arg1 Alecto 1165:1170 Pteropus Alecto 1156:1170 Pteropus Alecto 1156:1170 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 56 theme close 1175:1179 arg1 homolog 1181:1187 a close homolog 1173:1187 a close homolog of hZIP4 1173:1196 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 57 theme reduced 1244:1250 arg1 stability 1260:1268 reduced thermal stability 1244:1268 reduced thermal stability 1244:1268 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 3 58 theme conserved 453:461 arg1 machinery 492:500 the conserved transmembrane zinc transport machinery 449:500 the conserved transmembrane zinc transport machinery 449:500 These mutations occur not only in the conserved transmembrane zinc transport machinery, but also in the extracellular domain (ECD) of hZIP4, which is only present in a fraction of mammalian ZIPs. 33837739 7 59 theme thermal 1252:1258 arg1 stability 1260:1268 reduced thermal stability 1244:1268 reduced thermal stability 1244:1268 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 2 60 theme acrodermatitis 380:393 arg1 AE 410:411 AE 410:411 AE 410:411 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 2 60 theme acrodermatitis 380:393 arg1 disorder 370:377 a life-threatening autosomal recessive disorder 331:377 a life-threatening autosomal recessive disorder 331:377 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 2 60 theme acrodermatitis 380:393 arg1 enteropathica 395:407 acrodermatitis enteropathica 380:407 acrodermatitis enteropathica (AE) 380:412 Loss-of-function mutations of human ZIP4 (hZIP4) drastically reduce zinc absorption, causing a life-threatening autosomal recessive disorder, acrodermatitis enteropathica (AE). 33837739 4 61 theme ECD 632:634 arg1 mutations 636:644 these AE-causing ECD mutations 615:644 these AE-causing ECD mutations 615:644 How these AE-causing ECD mutations lead to ZIP4 malfunction has not be fully clarified. 33837739 7 62 theme corresponding 1092:1104 arg1 mutations 1106:1114 the corresponding mutations 1088:1114 the corresponding mutations 1088:1114 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 7 63 theme intracellular 1297:1309 arg1 mistrafficking 1311:1324 intracellular mistrafficking 1297:1324 intracellular mistrafficking of the AE-associated variants 1297:1354 When the corresponding mutations were introduced in the ECD of ZIP4 from Pteropus Alecto, a close homolog of hZIP4, the variants exhibited structural defects or reduced thermal stability, which likely accounts for intracellular mistrafficking of the AE-associated variants and as such a total loss of zinc uptake activity. 33837739 1 64 theme protein 162:168 arg1 family 188:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 the Zrt-/Irt-like protein (ZIP) transporter family 144:193 ZIP4 is a representative member of the Zrt-/Irt-like protein (ZIP) transporter family and responsible for zinc uptake from diet. 18657508 4 0 theme domains 692:698 arg1 structures 663:672 structures 663:672 structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity 663:836 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 3 1 theme CD47 513:516 arg1 face 505:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 5 2 from detail 935:940 arg1 family 898:903 the SIRP family 889:903 the SIRP family of paired receptors in atomic detail 889:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 6 3 theme SIRPalpha 955:963 arg1 polymorphisms 965:977 SIRPalpha polymorphisms 955:977 SIRPalpha polymorphisms 955:977 Analysis of SIRPalpha polymorphisms suggests that these, as well as the activating SIRPs, may have evolved to counteract pathogen binding to the inhibitory SIRPalpha receptor. 18657508 4 4 theme reduced 821:827 arg1 affinity 829:836 negligible or reduced affinity 807:836 negligible or reduced affinity 807:836 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 5 theme N-terminal 681:690 arg1 SIRPbeta 713:720 SIRPbeta(2) 713:723 SIRPbeta(2) 713:723 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 5 theme N-terminal 681:690 arg1 SIRPgamma 730:738 SIRPgamma 730:738 SIRPgamma 730:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 5 theme N-terminal 681:690 arg1 domains 692:698 the N-terminal domains 677:698 the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma 677:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 5 theme N-terminal 681:690 arg1 SIRPbeta 703:710 SIRPbeta 703:710 SIRPbeta 703:710 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 5 6 from receptors 915:923 arg1 detail 935:940 atomic detail 928:940 atomic detail 928:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 4 7 theme SIRPgamma 730:738 arg1 SIRPbeta 713:720 SIRPbeta(2) 713:723 SIRPbeta(2) 713:723 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 7 theme SIRPgamma 730:738 arg1 SIRPgamma 730:738 SIRPgamma 730:738 SIRPgamma 730:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 7 theme SIRPgamma 730:738 arg1 domains 692:698 the N-terminal domains 677:698 the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma 677:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 7 theme SIRPgamma 730:738 arg1 SIRPbeta 703:710 SIRPbeta 703:710 SIRPbeta 703:710 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 8 with CD47 797:800 arg1 affinity 829:836 negligible or reduced affinity 807:836 negligible or reduced affinity 807:836 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 5 9 theme SIRP 893:896 arg1 family 898:903 the SIRP family 889:903 the SIRP family of paired receptors in atomic detail 889:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 6 10 theme SIRPalpha 1099:1107 arg1 receptor 1109:1116 the inhibitory SIRPalpha receptor 1084:1116 the inhibitory SIRPalpha receptor 1084:1116 Analysis of SIRPalpha polymorphisms suggests that these, as well as the activating SIRPs, may have evolved to counteract pathogen binding to the inhibitory SIRPalpha receptor. 18657508 2 11 theme crystallographic 278:293 arg1 structures 295:304 the high-resolution X-ray crystallographic structures 252:304 the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 252:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 3 12 theme interacting 493:503 arg1 face 505:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 2 13 theme X-ray 272:276 arg1 structures 295:304 the high-resolution X-ray crystallographic structures 252:304 the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 252:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 6 14 theme activating 1015:1024 arg1 SIRPs 1026:1030 the activating SIRPs 1011:1030 the activating SIRPs 1011:1030 Analysis of SIRPalpha polymorphisms suggests that these, as well as the activating SIRPs, may have evolved to counteract pathogen binding to the inhibitory SIRPalpha receptor. 18657508 3 15 theme unusual 470:476 arg1 face 505:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 6 16 theme pathogen 1064:1071 arg1 binding 1073:1079 pathogen binding 1064:1079 pathogen binding to the inhibitory SIRPalpha receptor 1064:1116 Analysis of SIRPalpha polymorphisms suggests that these, as well as the activating SIRPs, may have evolved to counteract pathogen binding to the inhibitory SIRPalpha receptor. 18657508 2 17 theme high-resolution 256:270 arg1 structures 295:304 the high-resolution X-ray crystallographic structures 252:304 the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 252:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 18 theme ligand-binding 396:409 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 18 theme ligand-binding 396:409 arg1 domain 411:416 the N-terminal ligand-binding domain 381:416 the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 381:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 0 19 theme receptor 7:14 arg1 specificity 16:26 Paired receptor specificity 0:26 Paired receptor specificity 0:26 Paired receptor specificity explained by structures of signal regulatory proteins alone and complexed with CD47. 18657508 2 20 theme superfamily 328:338 arg1 CD47 350:353 CD47 350:353 CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 350:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 20 theme superfamily 328:338 arg1 domain 340:345 the immunoglobulin superfamily domain 309:345 the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 309:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 3 21 from regions 617:623 arg1 SIRPalpha 628:636 SIRPalpha 628:636 SIRPalpha 628:636 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 2 22 theme N-terminal 385:394 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 22 theme N-terminal 385:394 arg1 domain 411:416 the N-terminal ligand-binding domain 381:416 the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 381:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 0 23 theme Paired 0:5 arg1 specificity 16:26 Paired receptor specificity 0:26 Paired receptor specificity 0:26 Paired receptor specificity explained by structures of signal regulatory proteins alone and complexed with CD47. 18657508 4 24 theme negligible 807:816 arg1 affinity 829:836 negligible or reduced affinity 807:836 negligible or reduced affinity 807:836 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 2 25 theme immunoglobulin 313:326 arg1 CD47 350:353 CD47 350:353 CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 350:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 25 theme immunoglobulin 313:326 arg1 domain 340:345 the immunoglobulin superfamily domain 309:345 the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 309:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 6 26 theme inhibitory 1088:1097 arg1 receptor 1109:1116 the inhibitory SIRPalpha receptor 1084:1116 the inhibitory SIRPalpha receptor 1084:1116 Analysis of SIRPalpha polymorphisms suggests that these, as well as the activating SIRPs, may have evolved to counteract pathogen binding to the inhibitory SIRPalpha receptor. 18657508 2 27 theme protein 439:445 arg1 SIRPalpha 454:462 SIRPalpha 454:462 SIRPalpha 454:462 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 27 theme protein 439:445 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 28 theme alpha 447:451 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 28 theme alpha 447:451 arg1 domain 411:416 the N-terminal ligand-binding domain 381:416 the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 381:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 3 29 theme N 534:534 arg1 terminus 536:543 the N terminus 530:543 the N terminus 530:543 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 5 30 theme paired 908:913 arg1 receptors 915:923 paired receptors 908:923 paired receptors in atomic detail 908:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 2 31 theme regulatory 428:437 arg1 SIRPalpha 454:462 SIRPalpha 454:462 SIRPalpha 454:462 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 31 theme regulatory 428:437 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 5 32 theme CD47 880:883 arg1 specificity 865:875 the specificity 861:875 the specificity of CD47 for the SIRP family of paired receptors in atomic detail 861:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 2 33 theme signal 421:426 arg1 SIRPalpha 454:462 SIRPalpha 454:462 SIRPalpha 454:462 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 33 theme signal 421:426 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 0 34 theme signal 55:60 arg1 proteins 73:80 signal regulatory proteins 55:80 signal regulatory proteins alone 55:86 Paired receptor specificity explained by structures of signal regulatory proteins alone and complexed with CD47. 18657508 5 35 theme atomic 928:933 arg1 detail 935:940 atomic detail 928:940 atomic detail 928:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 4 36 theme SIRPbeta 703:710 arg1 SIRPbeta 713:720 SIRPbeta(2) 713:723 SIRPbeta(2) 713:723 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 36 theme SIRPbeta 703:710 arg1 SIRPgamma 730:738 SIRPgamma 730:738 SIRPgamma 730:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 36 theme SIRPbeta 703:710 arg1 domains 692:698 the N-terminal domains 677:698 the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma 677:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 36 theme SIRPbeta 703:710 arg1 SIRPbeta 703:710 SIRPbeta 703:710 SIRPbeta 703:710 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 3 37 theme domain 573:578 arg1 end 562:564 the end 558:564 the end of the domain 558:578 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 2 38 theme CD47 350:353 arg1 CD47 350:353 CD47 350:353 CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 350:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 38 theme CD47 350:353 arg1 domain 340:345 the immunoglobulin superfamily domain 309:345 the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 309:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 4 39 dep structures 663:672 arg1 proteins 741:748 proteins 741:748 structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity 663:836 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 3 40 theme corresponding 603:615 arg1 regions 617:623 the corresponding regions 599:623 the corresponding regions in SIRPalpha 599:636 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 3 41 theme convoluted 482:491 arg1 face 505:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face 466:508 The unusual and convoluted interacting face of CD47, comprising the N terminus and loops at the end of the domain, intercalates with the corresponding regions in SIRPalpha. 18657508 1 42 theme myeloid 214:220 arg1 cells 233:237 myeloid and neural cells 214:237 myeloid and neural cells 214:237 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 5 43 theme receptors 915:923 arg1 family 898:903 the SIRP family 889:903 the SIRP family of paired receptors in atomic detail 889:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 2 44 theme domain 340:345 arg1 structures 295:304 the high-resolution X-ray crystallographic structures 252:304 the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 252:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 4 45 theme SIRPbeta 713:720 arg1 SIRPbeta 713:720 SIRPbeta(2) 713:723 SIRPbeta(2) 713:723 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 45 theme SIRPbeta 713:720 arg1 SIRPgamma 730:738 SIRPgamma 730:738 SIRPgamma 730:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 45 theme SIRPbeta 713:720 arg1 domains 692:698 the N-terminal domains 677:698 the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma 677:738 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 4 45 theme SIRPbeta 713:720 arg1 SIRPbeta 703:710 SIRPbeta 703:710 SIRPbeta 703:710 We have also determined structures of the N-terminal domains of SIRPbeta, SIRPbeta(2), and SIRPgamma; proteins that are closely related to SIRPalpha but bind CD47 with negligible or reduced affinity. 18657508 0 46 theme proteins 73:80 arg1 structures 41:50 structures 41:50 structures of signal regulatory proteins alone 41:86 Paired receptor specificity explained by structures of signal regulatory proteins alone and complexed with CD47. 18657508 2 47 with complex 368:374 arg1 alpha 447:451 signal regulatory protein alpha 421:451 signal regulatory protein alpha (SIRPalpha) 421:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 2 47 with complex 368:374 arg1 domain 411:416 the N-terminal ligand-binding domain 381:416 the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha) 381:463 We describe the high-resolution X-ray crystallographic structures of the immunoglobulin superfamily domain of CD47 alone and in complex with the N-terminal ligand-binding domain of signal regulatory protein alpha (SIRPalpha). 18657508 1 48 theme self 185:188 arg1 marker 175:180 a marker 173:180 a marker of self 173:188 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 1 49 theme distributed 130:140 arg1 CD47 113:116 CD47 113:116 CD47 113:116 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 1 49 theme distributed 130:140 arg1 protein 155:161 a widely distributed cell-surface protein 121:161 a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells 121:237 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 5 50 from family 898:903 arg1 detail 935:940 atomic detail 928:940 atomic detail 928:940 These results explain the specificity of CD47 for the SIRP family of paired receptors in atomic detail. 18657508 1 51 theme neural 226:231 arg1 cells 233:237 myeloid and neural cells 214:237 myeloid and neural cells 214:237 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 0 52 theme regulatory 62:71 arg1 proteins 73:80 signal regulatory proteins 55:80 signal regulatory proteins alone 55:86 Paired receptor specificity explained by structures of signal regulatory proteins alone and complexed with CD47. 18657508 6 53 theme polymorphisms 965:977 arg1 Analysis 943:950 Analysis 943:950 Analysis of SIRPalpha polymorphisms 943:977 Analysis of SIRPalpha polymorphisms suggests that these, as well as the activating SIRPs, may have evolved to counteract pathogen binding to the inhibitory SIRPalpha receptor. 18657508 1 54 theme cell-surface 142:153 arg1 CD47 113:116 CD47 113:116 CD47 113:116 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 1 54 theme cell-surface 142:153 arg1 protein 155:161 a widely distributed cell-surface protein 121:161 a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells 121:237 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 18657508 1 55 theme cells 233:237 arg1 interactions 198:209 interactions 198:209 interactions of myeloid and neural cells 198:237 CD47 is a widely distributed cell-surface protein that acts a marker of self through interactions of myeloid and neural cells. 28775322 7 0 theme catalytic 1582:1590 arg1 mechanism 1592:1600 catalytic mechanism 1582:1600 catalytic mechanism 1582:1600 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 6 1 theme signaling 1381:1389 arg1 receptors 1391:1399 other signaling receptors 1375:1399 other signaling receptors 1375:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 2 theme receptors 1391:1399 arg1 domains 1354:1360 the EGF-like domains 1341:1360 the EGF-like domains of Notch and other signaling receptors 1341:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 2 theme receptors 1391:1399 arg1 receptors 1391:1399 other signaling receptors 1375:1399 other signaling receptors 1375:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 2 theme receptors 1391:1399 arg1 Notch 1365:1369 Notch 1365:1369 Notch 1365:1369 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 1 3 theme growth 196:201 arg1 domains 226:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 7 4 theme human 1443:1447 arg1 POGLUT1 1449:1455 human POGLUT1 1443:1455 human POGLUT1 1443:1455 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 7 5 theme donor 1508:1512 arg1 substrates 1514:1523 donor substrates 1508:1523 donor substrates 1508:1523 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 7 6 theme substrate 1556:1564 arg1 specificity 1566:1576 the enzyme's substrate specificity 1543:1576 the enzyme's substrate specificity 1543:1576 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 6 7 theme other 1375:1379 arg1 receptors 1391:1399 other signaling receptors 1375:1399 other signaling receptors 1375:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 1 8 theme factor-like 203:213 arg1 domains 226:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 6 9 theme Protein 1165:1171 arg1 O-glucosyltransferase 1173:1193 Protein O-glucosyltransferase 1 1165:1195 Protein O-glucosyltransferase 1 1165:1195 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 5 10 theme broader 1070:1076 arg1 range 1078:1082 a much broader range 1063:1082 a much broader range of EGF-like domain substrates 1063:1112 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 5 11 theme EGF-like 1087:1094 arg1 substrates 1103:1112 EGF-like domain substrates 1087:1112 EGF-like domain substrates 1087:1112 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 2 12 theme EGF-like 339:346 arg1 domains 348:354 folded EGF-like domains 332:354 folded EGF-like domains 332:354 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 7 13 from structure 1430:1438 arg1 complexes 1460:1468 complexes 1460:1468 complexes with 3 different EGF-like domains and donor substrates 1460:1523 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 6 14 located found 1246:1250 arg2 type 1241:1244 the type 1237:1244 the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1237:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 14 located found 1246:1250 arg1 Notch.POGLUT1 1255:1267 Notch.POGLUT1 1255:1267 Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1255:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 15 theme type 1241:1244 arg1 domains 1226:1232 EGF-like domains 1217:1232 EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1217:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 15 theme type 1241:1244 arg1 type 1241:1244 the type 1237:1244 the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1237:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 3 16 theme Protein 586:592 arg1 complexes 636:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 2 17 from residues 388:395 arg1 motif 413:417 the C1XSXPC2 motif 400:417 the C1XSXPC2 motif 400:417 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 2 18 theme folded 332:337 arg1 domains 348:354 folded EGF-like domains 332:354 folded EGF-like domains 332:354 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 3 19 theme substrate 524:532 arg1 analogs 534:540 donor substrate analogs 518:540 donor substrate analogs 518:540 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 6 20 theme EGF-like 1217:1224 arg1 domains 1226:1232 EGF-like domains 1217:1232 EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1217:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 20 theme EGF-like 1217:1224 arg1 type 1241:1244 the type 1237:1244 the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1237:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 7 21 theme different 1477:1485 arg1 domains 1496:1502 3 different EGF-like domains 1475:1502 3 different EGF-like domains 1475:1502 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 1 22 theme EGF-like 216:223 arg1 domains 226:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 5 23 theme domain 1096:1101 arg1 substrates 1103:1112 EGF-like domain substrates 1087:1112 EGF-like domain substrates 1087:1112 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 6 24 theme EGF-like 1345:1352 arg1 domains 1354:1360 the EGF-like domains 1341:1360 the EGF-like domains of Notch and other signaling receptors 1341:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 24 theme EGF-like 1345:1352 arg1 receptors 1391:1399 other signaling receptors 1375:1399 other signaling receptors 1375:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 24 theme EGF-like 1345:1352 arg1 Notch 1365:1369 Notch 1365:1369 Notch 1365:1369 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 3 25 theme substrate/product 618:634 arg1 complexes 636:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 0 26 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 0:115 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 28775322 4 27 theme folded 788:793 arg1 domains 804:810 folded EGF-like domains 788:810 folded EGF-like domains 788:810 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 3 28 theme complexes 636:644 arg1 structures 566:575 the structures 562:575 the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties 562:713 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 4 29 theme serine 830:835 arg1 specificity 837:847 its serine specificity 826:847 its serine specificity 826:847 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 1 30 theme domains 226:232 arg1 glucosylation 163:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation 117:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains 117:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 0 31 theme Notch 20:24 arg1 O-glucosylation 26:40 Notch O-glucosylation 20:40 Notch O-glucosylation 20:40 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 28775322 4 32 theme local 871:875 arg1 states 892:897 two distinct local conformational states 858:897 two distinct local conformational states 858:897 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 5 33 theme Protein 993:999 arg1 O-glucosyltransferase 1001:1021 Protein O-glucosyltransferase 1 993:1023 Protein O-glucosyltransferase 1 993:1023 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 3 34 theme O-glucosyltransferase 594:614 arg1 complexes 636:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 5 35 contain possesses 1025:1033 arg1 O-glucosyltransferase 1001:1021 Protein O-glucosyltransferase 1 993:1023 Protein O-glucosyltransferase 1 993:1023 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 5 35 contain possesses 1025:1033 arg2 potential 1039:1047 the potential to xylosylate a much broader range of EGF-like domain substrates 1035:1112 the potential to xylosylate a much broader range of EGF-like domain substrates 1035:1112 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 2 36 theme donor 454:458 arg1 specificity 470:480 an uncommon dual donor substrate specificity 437:480 an uncommon dual donor substrate specificity 437:480 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 2 37 theme substrate 460:468 arg1 specificity 470:480 an uncommon dual donor substrate specificity 437:480 an uncommon dual donor substrate specificity 437:480 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 1 38 theme Protein 117:123 arg1 glucosylation 163:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation 117:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains 117:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 2 39 theme dual 449:452 arg1 specificity 470:480 an uncommon dual donor substrate specificity 437:480 an uncommon dual donor substrate specificity 437:480 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 2 40 theme serine 381:386 arg1 residues 388:395 serine residues 381:395 serine residues in the C1XSXPC2 motif 381:417 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 2 41 theme uncommon 440:447 arg1 specificity 470:480 an uncommon dual donor substrate specificity 437:480 an uncommon dual donor substrate specificity 437:480 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 3 42 theme mechanistic 659:669 arg1 insight 671:677 mechanistic insight 659:677 mechanistic insight into the basis for these properties 659:713 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 1 43 theme important 243:251 arg1 role 253:256 an important role 240:256 an important role 240:256 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 0 44 theme O-glucosylation 26:40 arg1 basis 11:15 Structural basis 0:15 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 0:115 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 28775322 4 45 theme conformational 877:890 arg1 states 892:897 two distinct local conformational states 858:897 two distinct local conformational states 858:897 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 3 46 theme EGF-like 497:504 arg1 domains 506:512 several EGF-like domains 489:512 several EGF-like domains 489:512 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 7 47 with complexes 1460:1468 arg1 domains 1496:1502 3 different EGF-like domains 1475:1502 3 different EGF-like domains 1475:1502 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 7 47 with complexes 1460:1468 arg1 substrates 1514:1523 donor substrates 1508:1523 donor substrates 1508:1523 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 2 48 contain possesses 427:435 arg1 it 424:425 it 424:425 it 424:425 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 2 48 contain possesses 427:435 arg2 specificity 470:480 an uncommon dual donor substrate specificity 437:480 an uncommon dual donor substrate specificity 437:480 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 5 49 theme substrates 1103:1112 arg1 range 1078:1082 a much broader range 1063:1082 a much broader range of EGF-like domain substrates 1063:1112 We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. 28775322 1 50 theme O-glucosyltransferase 125:145 arg1 glucosylation 163:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation 117:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains 117:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 0 51 theme O-xylosylation 46:59 arg1 basis 11:15 Structural basis 0:15 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 0:115 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 28775322 6 52 theme Notch 1365:1369 arg1 domains 1354:1360 the EGF-like domains 1341:1360 the EGF-like domains of Notch and other signaling receptors 1341:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 52 theme Notch 1365:1369 arg1 receptors 1391:1399 other signaling receptors 1375:1399 other signaling receptors 1375:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 6 52 theme Notch 1365:1369 arg1 Notch 1365:1369 Notch 1365:1369 Notch 1365:1369 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 1 53 theme 1/Rumi-mediated 147:161 arg1 glucosylation 163:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation 117:175 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains 117:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 4 54 theme EGF-like 795:802 arg1 domains 804:810 folded EGF-like domains 788:810 folded EGF-like domains 788:810 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 4 55 theme distinct 862:869 arg1 states 892:897 two distinct local conformational states 858:897 two distinct local conformational states 858:897 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 7 56 theme POGLUT1 1449:1455 arg1 structure 1430:1438 the structure 1426:1438 the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates 1426:1523 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 4 57 theme Protein 738:744 arg1 O-glucosyltransferase 746:766 Protein O-glucosyltransferase 1 738:768 Protein O-glucosyltransferase 1's requirement for folded EGF-like domains 738:810 Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. 28775322 3 58 theme donor 518:522 arg1 analogs 534:540 donor substrate analogs 518:540 donor substrate analogs 518:540 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 7 59 theme EGF-like 1487:1494 arg1 domains 1496:1502 3 different EGF-like domains 1475:1502 3 different EGF-like domains 1475:1502 Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism. 28775322 2 60 gly glycosylate 369:379 arg1 residues 388:395 serine residues 381:395 serine residues in the C1XSXPC2 motif 381:417 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 3 61 theme human 580:584 arg1 complexes 636:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 human Protein O-glucosyltransferase 1 substrate/product complexes 580:644 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 1 62 theme epidermal 186:194 arg1 domains 226:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 1 63 theme Notch 261:265 arg1 signaling 267:275 Notch signaling 261:275 Notch signaling 261:275 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 6 64 dep Notch.POGLUT1 1255:1267 arg1 protein-O-glucosyltransferase 1274:1302 a protein-O-glucosyltransferase 1272:1302 a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors 1272:1399 Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. 28775322 3 65 theme several 489:495 arg1 domains 506:512 several EGF-like domains 489:512 several EGF-like domains 489:512 Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. 28775322 1 66 theme Notch 180:184 arg1 domains 226:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Notch epidermal growth factor-like (EGF-like) domains 180:232 Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. 28775322 0 67 theme mammalian 64:72 arg1 protein-O-glucosyltransferase 74:102 mammalian protein-O-glucosyltransferase 1 64:104 mammalian protein-O-glucosyltransferase 1 (POGLUT1) 64:114 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 28775322 0 67 theme mammalian 64:72 arg1 POGLUT1 107:113 POGLUT1 107:113 POGLUT1 107:113 Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1). 28775322 2 68 theme Protein 278:284 arg1 O-glucosyltransferase 286:306 Protein O-glucosyltransferase 1 278:308 Protein O-glucosyltransferase 1 278:308 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 28775322 2 69 theme C1XSXPC2 404:411 arg1 motif 413:417 the C1XSXPC2 motif 400:417 the C1XSXPC2 motif 400:417 Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. 9342320 4 0 contain contains 676:683 arg1 IFN-beta 667:674 human IFN-beta 661:674 human IFN-beta 661:674 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 4 0 contain contains 676:683 arg2 site 700:703 a zinc-binding site 685:703 a zinc-binding site 685:703 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 4 0 contain contains 676:683 arg1 interface 712:720 the interface 708:720 the interface of the two molecules in the asymmetric unit 708:764 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 4 0 contain contains 676:683 arg2 IFN-beta 667:674 human IFN-beta 661:674 human IFN-beta 661:674 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 1 1 theme diverse 126:132 arg1 activities 145:154 diverse biological activities 126:154 diverse biological activities 126:154 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 2 theme receptor 216:223 arg1 system 225:230 the same receptor system 207:230 the same receptor system 207:230 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 2 3 theme molecular 439:447 arg1 replacement 449:459 molecular replacement 439:459 molecular replacement 439:459 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 5 4 theme IFN-alpha2b 859:869 arg1 dimer 871:875 the human IFN-alpha2b dimer 849:875 the human IFN-alpha2b dimer 849:875 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 1 5 theme biological 134:143 arg1 activities 145:154 diverse biological activities 126:154 diverse biological activities 126:154 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 7 6 from role 1143:1146 arg1 assembly 1200:1207 the conformational assembly 1181:1207 the conformational assembly of IFN receptor components 1181:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 7 7 theme ligand-ligand 1151:1163 arg1 interactions 1165:1176 ligand-ligand interactions 1151:1176 ligand-ligand interactions 1151:1176 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 5 8 theme contact 957:963 arg1 surfaces 965:972 contact surfaces 957:972 contact surfaces from opposite sides of the molecule 957:1008 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 4 9 theme human 661:665 arg1 IFN-beta 667:674 human IFN-beta 661:674 human IFN-beta 661:674 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 4 10 theme molecules 733:741 arg1 interface 712:720 the interface 708:720 the interface of the two molecules in the asymmetric unit 708:764 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 6 11 theme specific 1091:1098 arg1 residues 1108:1115 specific exposed residues 1091:1115 specific exposed residues 1091:1115 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 2 12 theme basis 285:289 arg1 understanding 253:265 a better understanding 244:265 a better understanding of the structural basis for the different activities of alpha and beta IFNs 244:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 2 13 theme better 246:251 arg1 understanding 253:265 a better understanding 244:265 a better understanding of the structural basis for the different activities of alpha and beta IFNs 244:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 7 14 theme IFN 1212:1214 arg1 components 1225:1234 IFN receptor components 1212:1234 IFN receptor components 1212:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 2 15 theme alpha 323:327 arg1 IFNs 338:341 alpha and beta IFNs 323:341 alpha and beta IFNs 323:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 5 16 theme homologous 887:896 arg1 surfaces 898:905 homologous surfaces 887:905 homologous surfaces 887:905 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 3 17 theme similar 489:495 arg1 fold 484:487 a fold 482:487 a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b 482:584 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 1 18 contain have 121:124 arg2 activities 145:154 diverse biological activities 126:154 diverse biological activities 126:154 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 18 contain have 121:124 arg1 cytokines 106:114 helical cytokines 98:114 helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system 98:230 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 18 contain have 121:124 arg1 interferons 75:85 Type I interferons 68:85 Type I interferons (IFNs) 68:92 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 4 19 theme relevance 802:810 arg1 question 779:786 the question 775:786 the question of functional relevance for IFN-beta dimers 775:830 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 1 20 theme Type 68:71 arg1 interferons 75:85 Type I interferons 68:85 Type I interferons (IFNs) 68:92 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 20 theme Type 68:71 arg1 cytokines 106:114 helical cytokines 98:114 helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system 98:230 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 20 theme Type 68:71 arg1 IFNs 88:91 IFNs 88:91 IFNs 88:91 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 6 21 theme structure 1032:1040 arg1 relevance 1015:1023 The relevance 1011:1023 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues 1011:1115 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 0 22 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human interferon beta at 2.2-A resolution 0:65 The crystal structure of human interferon beta at 2.2-A resolution. 9342320 7 23 theme components 1225:1234 arg1 assembly 1200:1207 the conformational assembly 1181:1207 the conformational assembly of IFN receptor components 1181:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 1 24 theme I 73:73 arg1 interferons 75:85 Type I interferons 68:85 Type I interferons (IFNs) 68:92 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 24 theme I 73:73 arg1 cytokines 106:114 helical cytokines 98:114 helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system 98:230 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 24 theme I 73:73 arg1 IFNs 88:91 IFNs 88:91 IFNs 88:91 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 2 25 theme human 401:405 arg1 IFN-beta 407:414 glycosylated human IFN-beta 388:414 glycosylated human IFN-beta 388:414 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 4 26 theme IFN-beta 816:823 arg1 dimers 825:830 IFN-beta dimers 816:830 IFN-beta dimers 816:830 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 4 27 from interface 712:720 arg1 unit 761:764 the asymmetric unit 746:764 the asymmetric unit 746:764 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 2 28 theme structural 274:283 arg1 basis 285:289 the structural basis 270:289 the structural basis for the different activities of alpha and beta IFNs 270:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 2 29 theme IFNs 338:341 arg1 activities 309:318 the different activities 295:318 the different activities of alpha and beta IFNs 295:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 2 30 theme glycosylated 388:399 arg1 IFN-beta 407:414 glycosylated human IFN-beta 388:414 glycosylated human IFN-beta 388:414 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 3 31 theme determined 523:532 arg1 structures 534:543 the previously determined structures 508:543 the previously determined structures of murine IFN-beta and human IFN-alpha2b 508:584 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 6 32 theme exposed 1100:1106 arg1 residues 1108:1115 specific exposed residues 1091:1115 specific exposed residues 1091:1115 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 5 33 theme opposite 979:986 arg1 sides 988:992 opposite sides 979:992 opposite sides of the molecule 979:1008 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 6 34 theme mutations 1066:1074 arg1 effects 1049:1055 the effects 1045:1055 the effects of point mutations in IFN-beta at specific exposed residues 1045:1115 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 3 35 theme structural 616:625 arg1 features 627:634 several distinct structural features 599:634 several distinct structural features 599:634 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 5 36 from sides 988:992 arg1 surfaces 965:972 contact surfaces 957:972 contact surfaces from opposite sides of the molecule 957:1008 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 3 37 theme several 599:605 arg1 features 627:634 several distinct structural features 599:634 several distinct structural features 599:634 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 2 38 theme crystal 367:373 arg1 structure 375:383 the crystal structure 363:383 the crystal structure of glycosylated human IFN-beta 363:414 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 0 39 theme interferon 31:40 arg1 beta 42:45 human interferon beta 25:45 human interferon beta 25:45 The crystal structure of human interferon beta at 2.2-A resolution. 9342320 7 40 theme receptor 1216:1223 arg1 components 1225:1234 IFN receptor components 1212:1234 IFN receptor components 1212:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 3 41 theme IFN-beta 555:562 arg1 structures 534:543 the previously determined structures 508:543 the previously determined structures of murine IFN-beta and human IFN-alpha2b 508:584 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 4 42 theme human 642:646 arg1 IFN-alpha2b 648:658 human IFN-alpha2b 642:658 human IFN-alpha2b 642:658 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 0 43 theme human 25:29 arg1 beta 42:45 human interferon beta 25:45 human interferon beta 25:45 The crystal structure of human interferon beta at 2.2-A resolution. 9342320 0 44 from resolution 56:65 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human interferon beta at 2.2-A resolution 0:65 The crystal structure of human interferon beta at 2.2-A resolution. 9342320 2 45 theme 2.2-A 419:423 arg1 resolution 425:434 2.2-A resolution 419:434 2.2-A resolution 419:434 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 3 46 theme distinct 607:614 arg1 features 627:634 several distinct structural features 599:634 several distinct structural features 599:634 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 4 47 theme functional 791:800 arg1 relevance 802:810 functional relevance 791:810 functional relevance for IFN-beta dimers 791:830 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 6 48 theme point 1060:1064 arg1 mutations 1066:1074 point mutations 1060:1074 point mutations in IFN-beta 1060:1086 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 4 49 theme zinc-binding 687:698 arg1 site 700:703 a zinc-binding site 685:703 a zinc-binding site 685:703 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 0 50 theme beta 42:45 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human interferon beta at 2.2-A resolution 0:65 The crystal structure of human interferon beta at 2.2-A resolution. 9342320 5 51 theme molecule 1001:1008 arg1 sides 988:992 opposite sides 979:992 opposite sides of the molecule 979:1008 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 3 52 theme murine 548:553 arg1 IFN-beta 555:562 murine IFN-beta 548:562 murine IFN-beta 548:562 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 7 53 theme interactions 1165:1176 arg1 role 1143:1146 A potential role 1131:1146 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components 1131:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 2 54 theme different 299:307 arg1 activities 309:318 the different activities 295:318 the different activities of alpha and beta IFNs 295:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 3 55 theme IFN-alpha2b 574:584 arg1 structures 534:543 the previously determined structures 508:543 the previously determined structures of murine IFN-beta and human IFN-alpha2b 508:584 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 6 56 from mutations 1066:1074 arg1 IFN-beta 1079:1086 IFN-beta 1079:1086 IFN-beta 1079:1086 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 6 57 from IFN-beta 1079:1086 arg1 effects 1049:1055 the effects 1045:1055 the effects of point mutations in IFN-beta at specific exposed residues 1045:1115 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 1 58 theme helical 98:104 arg1 interferons 75:85 Type I interferons 68:85 Type I interferons (IFNs) 68:92 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 1 58 theme helical 98:104 arg1 cytokines 106:114 helical cytokines 98:114 helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system 98:230 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 9342320 6 59 from residues 1108:1115 arg1 effects 1049:1055 the effects 1045:1055 the effects of point mutations in IFN-beta at specific exposed residues 1045:1115 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 4 60 theme asymmetric 750:759 arg1 unit 761:764 the asymmetric unit 746:764 the asymmetric unit 746:764 Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. 9342320 6 61 from effects 1049:1055 arg1 IFN-beta 1079:1086 IFN-beta 1079:1086 IFN-beta 1079:1086 The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. 9342320 5 62 theme human 853:857 arg1 dimer 871:875 the human IFN-alpha2b dimer 849:875 the human IFN-alpha2b dimer 849:875 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 7 63 theme conformational 1185:1198 arg1 assembly 1200:1207 the conformational assembly 1181:1207 the conformational assembly of IFN receptor components 1181:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 3 64 theme human 568:572 arg1 IFN-alpha2b 574:584 human IFN-alpha2b 568:584 human IFN-alpha2b 568:584 The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. 9342320 2 65 theme beta 333:336 arg1 IFNs 338:341 alpha and beta IFNs 323:341 alpha and beta IFNs 323:341 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 0 66 theme 2.2-A 50:54 arg1 resolution 56:65 2.2-A resolution 50:65 2.2-A resolution 50:65 The crystal structure of human interferon beta at 2.2-A resolution. 9342320 5 67 theme human 927:931 arg1 IFN-beta 933:940 human IFN-beta 927:940 human IFN-beta 927:940 However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. 9342320 2 68 gly glycosylated 388:399 arg1 IFN-beta 407:414 glycosylated human IFN-beta 388:414 glycosylated human IFN-beta 388:414 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 2 69 theme IFN-beta 407:414 arg1 structure 375:383 the crystal structure 363:383 the crystal structure of glycosylated human IFN-beta 363:414 To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. 9342320 7 70 theme potential 1133:1141 arg1 role 1143:1146 A potential role 1131:1146 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components 1131:1234 A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. 9342320 1 71 theme same 211:214 arg1 system 225:230 the same receptor system 207:230 the same receptor system 207:230 Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. 6785754 0 0 theme delta 57:61 arg1 chain 63:67 a human delta chain 49:67 a human delta chain 49:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 6 1 theme immunoglobulins 754:768 arg1 structure 741:749 the common framework structure 720:749 the common framework structure of immunoglobulins 720:768 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 1 2 with proteolysis 174:184 arg1 trypsin 191:197 trypsin 191:197 trypsin of an intact myeloma IgD protein (NIG-65) 191:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 1 3 theme fragment 117:124 arg1 sequence 94:101 The complete amino acid sequence 70:101 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 70:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 0 4 theme human 51:55 arg1 chain 63:67 a human delta chain 49:67 a human delta chain 49:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 6 5 theme heavy 902:906 arg1 chains 908:913 other heavy chains 896:913 other heavy chains 896:913 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 9 6 theme moderate 1289:1296 arg1 degree 1298:1303 a moderate degree 1287:1303 a moderate degree of internal homology 1287:1324 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 9 7 theme homology 1317:1324 arg1 degree 1298:1303 a moderate degree 1287:1303 a moderate degree of internal homology 1287:1324 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 9 8 theme immunoglobulin 1271:1284 arg1 class 1262:1266 each class 1257:1266 each class of immunoglobulin 1257:1284 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 8 9 theme prominent 1174:1182 arg1 gaps 1184:1187 three prominent gaps 1168:1187 three prominent gaps by which each domain can be divided into two homologous halves 1168:1250 The comparison has also shown that there are three prominent gaps by which each domain can be divided into two homologous halves. 6785754 2 10 contain contains 275:282 arg1 fragment 266:273 The fragment 262:273 The fragment 262:273 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 2 10 contain contains 275:282 arg2 residues 299:306 226 amino acid residues 284:306 226 amino acid residues 284:306 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 4 11 theme other 557:561 arg1 classes 563:569 the other classes 553:569 the other classes 553:569 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 1 12 theme intact 205:210 arg1 NIG-65 233:238 NIG-65 233:238 NIG-65 233:238 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 1 12 theme intact 205:210 arg1 protein 224:230 an intact myeloma IgD protein 202:230 an intact myeloma IgD protein (NIG-65) 202:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 0 13 theme chain 63:67 arg1 region 39:44 the Fc region 32:44 the Fc region of a human delta chain 32:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 6 14 theme framework 731:739 arg1 structure 741:749 the common framework structure 720:749 the common framework structure of immunoglobulins 720:768 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 7 15 theme Fc 955:956 arg1 chain 979:983 the delta chain 969:983 the delta chain 969:983 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 7 15 theme Fc 955:956 arg1 domains 958:964 the two Fc domains 947:964 the two Fc domains of the delta chain 947:983 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 7 16 from branch 1020:1025 arg1 topology 1030:1037 topology 1030:1037 topology 1030:1037 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 2 17 theme amino 288:292 arg1 residues 299:306 226 amino acid residues 284:306 226 amino acid residues 284:306 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 1 18 theme myeloma 212:218 arg1 NIG-65 233:238 NIG-65 233:238 NIG-65 233:238 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 1 18 theme myeloma 212:218 arg1 protein 224:230 an intact myeloma IgD protein 202:230 an intact myeloma IgD protein (NIG-65) 202:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 7 19 theme independent 1008:1018 arg1 branch 1020:1025 an independent branch 1005:1025 an independent branch in topology 1005:1037 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 6 20 theme common 724:729 arg1 structure 741:749 the common framework structure 720:749 the common framework structure of immunoglobulins 720:768 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 21 theme other 896:900 arg1 chains 908:913 other heavy chains 896:913 other heavy chains 896:913 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 22 theme counterpart 874:884 arg1 domain 886:891 the counterpart domain 870:891 the counterpart domain of other heavy chains 870:913 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 22 theme counterpart 874:884 arg1 chains 908:913 other heavy chains 896:913 other heavy chains 896:913 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 1 23 theme Fc 137:138 arg1 t 147:147 t 147:147 t 147:147 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 1 23 theme Fc 137:138 arg1 delta 140:144 Fc delta 137:144 Fc delta (t) 137:148 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 4 24 theme delta 499:503 arg1 chain 505:509 the delta chain 495:509 the delta chain 495:509 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 10 25 theme genes 1636:1640 arg1 evolution 1619:1627 evolution 1619:1627 evolution 1619:1627 Based on this observation together with sequence comparisons, a possible genetic mechanism is proposed for the origin and evolution of the genes for immunoglobulin domains. 6785754 10 25 theme genes 1636:1640 arg1 origin 1608:1613 origin 1608:1613 origin 1608:1613 Based on this observation together with sequence comparisons, a possible genetic mechanism is proposed for the origin and evolution of the genes for immunoglobulin domains. 6785754 9 26 theme first 1345:1349 arg1 half 1351:1354 the first half 1341:1354 the first half 1341:1354 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 9 27 theme second 1364:1369 arg1 half 1371:1374 the second half 1360:1374 the second half of each domain of the Fc 1360:1399 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 0 28 theme amino 9:13 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Fc region of a human delta chain 0:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 1 29 dep fragment 117:124 arg1 obtained 154:161 obtained 154:161 obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 154:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 1 29 dep fragment 117:124 arg1 designated 126:135 designated 126:135 designated Fc delta (t) 126:148 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 0 30 theme Complete 0:7 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Fc region of a human delta chain 0:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 1 31 theme IgD 220:222 arg1 NIG-65 233:238 NIG-65 233:238 NIG-65 233:238 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 1 31 theme IgD 220:222 arg1 protein 224:230 an intact myeloma IgD protein 202:230 an intact myeloma IgD protein (NIG-65) 202:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 7 32 with paired 1086:1091 arg1 counterparts 1109:1120 their counterparts 1103:1120 their counterparts 1103:1120 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 3 33 contain has 368:370 arg2 oligosaccharides 390:405 three glucosamine oligosaccharides 372:405 three glucosamine oligosaccharides 372:405 It has three glucosamine oligosaccharides at asparagine residues 68, 159, and 210. 6785754 3 33 contain has 368:370 arg1 It 365:366 It 365:366 It 365:366 It has three glucosamine oligosaccharides at asparagine residues 68, 159, and 210. 6785754 1 34 theme protein 224:230 arg1 trypsin 191:197 trypsin 191:197 trypsin of an intact myeloma IgD protein (NIG-65) 191:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 7 35 theme Such 916:919 arg1 comparison 921:930 Such comparison 916:930 Such comparison 916:930 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 3 36 theme glucosamine 378:388 arg1 oligosaccharides 390:405 three glucosamine oligosaccharides 372:405 three glucosamine oligosaccharides 372:405 It has three glucosamine oligosaccharides at asparagine residues 68, 159, and 210. 6785754 0 37 theme acid 15:18 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Fc region of a human delta chain 0:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 8 38 theme homologous 1234:1243 arg1 halves 1245:1250 two homologous halves 1230:1250 two homologous halves 1230:1250 The comparison has also shown that there are three prominent gaps by which each domain can be divided into two homologous halves. 6785754 1 39 theme complete 74:81 arg1 sequence 94:101 The complete amino acid sequence 70:101 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 70:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 2 40 theme 32,000 338:343 arg1 weight 328:333 a molecular weight 316:333 a molecular weight of 32,000 per monomeric unit 316:362 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 7 41 theme chain 979:983 arg1 chain 979:983 the delta chain 969:983 the delta chain 969:983 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 7 41 theme chain 979:983 arg1 domains 958:964 the two Fc domains 947:964 the two Fc domains of the delta chain 947:983 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 6 42 theme individual 793:802 arg1 characteristics 804:818 many individual characteristics 788:818 many individual characteristics 788:818 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 2 43 contain has 312:314 arg1 fragment 266:273 The fragment 262:273 The fragment 262:273 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 2 43 contain has 312:314 arg2 weight 328:333 a molecular weight 316:333 a molecular weight of 32,000 per monomeric unit 316:362 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 4 44 from position 534:541 arg1 any 546:548 any 546:548 any 546:548 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 9 45 theme internal 1308:1315 arg1 homology 1317:1324 internal homology 1308:1324 internal homology 1308:1324 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 2 46 theme molecular 318:326 arg1 weight 328:333 a molecular weight 316:333 a molecular weight of 32,000 per monomeric unit 316:362 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 6 47 theme many 788:791 arg1 characteristics 804:818 many individual characteristics 788:818 many individual characteristics 788:818 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 9 48 theme Fc 1398:1399 arg1 domain 1384:1389 each domain 1379:1389 each domain of the Fc 1379:1399 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 9 48 theme Fc 1398:1399 arg1 Fc 1398:1399 the Fc 1394:1399 the Fc 1394:1399 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 0 49 theme region 39:44 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of the Fc region of a human delta chain 0:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 9 50 theme domain 1489:1494 arg1 size 1474:1477 the size 1470:1477 the size of a half domain 1470:1494 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 10 51 theme sequence 1537:1544 arg1 comparisons 1546:1556 sequence comparisons 1537:1556 sequence comparisons 1537:1556 Based on this observation together with sequence comparisons, a possible genetic mechanism is proposed for the origin and evolution of the genes for immunoglobulin domains. 6785754 1 52 theme amino 83:87 arg1 sequence 94:101 The complete amino acid sequence 70:101 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 70:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 0 53 theme Fc 36:37 arg1 region 39:44 the Fc region 32:44 the Fc region of a human delta chain 32:67 Complete amino acid sequence of the Fc region of a human delta chain. 6785754 4 54 theme these 451:455 arg1 characteristic 477:490 characteristic 477:490 characteristic of the delta chain 477:509 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 6 55 theme Fc 703:704 arg1 t 713:713 t 713:713 t 713:713 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 55 theme Fc 703:704 arg1 delta 706:710 the Fc delta 699:710 the Fc delta (t) 699:714 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 9 56 theme domain 1384:1389 arg1 half 1371:1374 the second half 1360:1374 the second half of each domain of the Fc 1360:1399 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 9 56 theme domain 1384:1389 arg1 half 1351:1354 the first half 1341:1354 the first half 1341:1354 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 9 57 theme primordial 1422:1431 arg1 gene 1433:1436 the primordial gene 1418:1436 the primordial gene 1418:1436 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 1 58 theme acid 89:92 arg1 sequence 94:101 The complete amino acid sequence 70:101 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 70:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 4 59 contain has 515:517 arg1 glucosamine-159 458:472 glucosamine-159 458:472 glucosamine-159 458:472 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 4 59 contain has 515:517 arg2 position 534:541 no counterpart position 519:541 no counterpart position in any of the other classes 519:569 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 5 60 theme other 579:583 arg1 hand 585:588 the other hand 575:588 the other hand 575:588 On the other hand, glucosamine-68 is shared by gamma, mu, and epsilon, and glucosamine-210 is shared by alpha and mu. 6785754 7 61 theme delta 973:977 arg1 chain 979:983 the delta chain 969:983 the delta chain 969:983 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 7 62 theme Fc 1071:1072 arg1 domains 1074:1080 the Fc domains 1067:1080 the Fc domains 1067:1080 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 1 63 theme limited 166:172 arg1 proteolysis 174:184 limited proteolysis 166:184 limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 166:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 6 64 theme chains 908:913 arg1 domain 886:891 the counterpart domain 870:891 the counterpart domain of other heavy chains 870:913 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 64 theme chains 908:913 arg1 chains 908:913 other heavy chains 896:913 other heavy chains 896:913 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 2 65 theme acid 294:297 arg1 residues 299:306 226 amino acid residues 284:306 226 amino acid residues 284:306 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 10 66 theme possible 1561:1568 arg1 mechanism 1578:1586 a possible genetic mechanism 1559:1586 a possible genetic mechanism 1559:1586 Based on this observation together with sequence comparisons, a possible genetic mechanism is proposed for the origin and evolution of the genes for immunoglobulin domains. 6785754 9 67 theme half 1484:1487 arg1 domain 1489:1494 a half domain 1482:1494 a half domain 1482:1494 For each class of immunoglobulin, a moderate degree of internal homology exists between the first half and the second half of each domain of the Fc, suggesting that the primordial gene may have coded for a unit about the size of a half domain. 6785754 6 68 contain has 784:786 arg2 characteristics 804:818 many individual characteristics 788:818 many individual characteristics 788:818 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 68 contain has 784:786 arg1 sequence 775:782 its sequence 771:782 its sequence 771:782 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 69 contain has 716:718 arg1 t 713:713 t 713:713 t 713:713 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 69 contain has 716:718 arg2 structure 741:749 the common framework structure 720:749 the common framework structure of immunoglobulins 720:768 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 6 69 contain has 716:718 arg1 delta 706:710 the Fc delta 699:710 the Fc delta (t) 699:714 Although the Fc delta (t) has the common framework structure of immunoglobulins, its sequence has many individual characteristics when its two domains are compared separately with the counterpart domain of other heavy chains. 6785754 3 70 theme asparagine 410:419 arg1 residues 421:428 asparagine residues 68, 159, and 210 410:445 asparagine residues 68, 159, and 210 410:445 It has three glucosamine oligosaccharides at asparagine residues 68, 159, and 210. 6785754 4 71 theme counterpart 522:532 arg1 position 534:541 no counterpart position 519:541 no counterpart position in any of the other classes 519:569 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 4 72 theme chain 505:509 arg1 characteristic 477:490 characteristic 477:490 characteristic of the delta chain 477:509 Of these, glucosamine-159 is characteristic of the delta chain and has no counterpart position in any of the other classes. 6785754 10 73 theme immunoglobulin 1646:1659 arg1 domains 1661:1667 immunoglobulin domains 1646:1667 immunoglobulin domains 1646:1667 Based on this observation together with sequence comparisons, a possible genetic mechanism is proposed for the origin and evolution of the genes for immunoglobulin domains. 6785754 2 74 theme monomeric 349:357 arg1 unit 359:362 monomeric unit 349:362 monomeric unit 349:362 The fragment contains 226 amino acid residues and has a molecular weight of 32,000 per monomeric unit. 6785754 1 75 theme Fc-like 109:115 arg1 fragment 117:124 an Fc-like fragment 106:124 an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) 106:239 The complete amino acid sequence of an Fc-like fragment designated Fc delta (t) and obtained by limited proteolysis with trypsin of an intact myeloma IgD protein (NIG-65) has been determined. 6785754 7 76 theme other 1052:1056 arg1 classes 1058:1064 all the other classes 1044:1064 all the other classes 1044:1064 Such comparison has shown that the two Fc domains of the delta chain should be placed in an independent branch in topology; for all the other classes, the Fc domains are paired well with their counterparts. 6785754 10 77 theme genetic 1570:1576 arg1 mechanism 1578:1586 a possible genetic mechanism 1559:1586 a possible genetic mechanism 1559:1586 Based on this observation together with sequence comparisons, a possible genetic mechanism is proposed for the origin and evolution of the genes for immunoglobulin domains. 32698188 3 0 theme neuronal 303:310 arg1 excitability 312:323 neuronal excitability 303:323 neuronal excitability linked to respiration, locomotion and circadian rhythm4-10 303:382 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 9 1 with domains 1084:1090 arg1 linkages 1108:1115 asymmetric linkages 1097:1115 asymmetric linkages to the pore 1097:1127 Unusual voltage-sensor domains with asymmetric linkages to the pore suggest mechanisms by which NALCN activity is modulated. 32698188 8 2 theme lateral 1033:1039 arg1 fenestrations 1046:1058 occluded lateral pore fenestrations 1024:1058 occluded lateral pore fenestrations 1024:1058 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 8 3 theme NALCN 960:964 arg1 pharmacology 944:955 The pharmacology 940:955 The pharmacology of NALCN 940:964 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 10 4 theme missense 1253:1260 arg1 mutations 1270:1278 missense patient mutations 1253:1278 missense patient mutations 1253:1278 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 9 5 theme NALCN 1157:1161 arg1 activity 1163:1170 NALCN activity 1157:1170 NALCN activity 1157:1170 Unusual voltage-sensor domains with asymmetric linkages to the pore suggest mechanisms by which NALCN activity is modulated. 32698188 8 6 theme occluded 1024:1031 arg1 fenestrations 1046:1058 occluded lateral pore fenestrations 1024:1058 occluded lateral pore fenestrations 1024:1058 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 6 7 theme member 820:825 arg1 FAM155A 830:836 FAM155A 830:836 FAM155A 830:836 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 7 theme member 820:825 arg1 A 827:827 155 member A 816:827 sequence similarity 155 member A (FAM155A) 796:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 4 8 theme severe 460:465 arg1 disorders 480:488 severe neurological disorders 460:488 severe neurological disorders 460:488 NALCN activity is under tight regulation11-14 and mutations in NALCN cause severe neurological disorders and early death15,16. 32698188 7 9 theme extracellular 857:869 arg1 dome 871:874 an extracellular dome 854:874 an extracellular dome that shields the ion-selectivity filter from neurotoxin attack 854:937 FAM155A forms an extracellular dome that shields the ion-selectivity filter from neurotoxin attack. 32698188 8 10 theme walled-off 993:1002 arg1 cavity 1012:1017 a walled-off central cavity 991:1017 a walled-off central cavity with occluded lateral pore fenestrations 991:1058 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 5 11 theme ion 603:605 arg1 aspects 566:572 fundamental aspects 554:572 fundamental aspects of channel assembly 554:592 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 11 theme ion 603:605 arg1 selectivity 607:617 ion selectivity 603:617 ion selectivity 603:617 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 7 12 theme neurotoxin 921:930 arg1 attack 932:937 neurotoxin attack 921:937 neurotoxin attack 921:937 FAM155A forms an extracellular dome that shields the ion-selectivity filter from neurotoxin attack. 32698188 8 13 with cavity 1012:1017 arg1 fenestrations 1046:1058 occluded lateral pore fenestrations 1024:1058 occluded lateral pore fenestrations 1024:1058 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 8 14 theme pore 1041:1044 arg1 fenestrations 1046:1058 occluded lateral pore fenestrations 1024:1058 occluded lateral pore fenestrations 1024:1058 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 4 15 theme tight 409:413 arg1 regulation11-14 415:429 tight regulation11-14 409:429 tight regulation11-14 409:429 NALCN activity is under tight regulation11-14 and mutations in NALCN cause severe neurological disorders and early death15,16. 32698188 11 16 theme foundation 1452:1461 arg1 physiology 1426:1435 the physiology 1422:1435 the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders 1422:1546 Our findings provide a framework to further study the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders. 32698188 10 17 theme pore 1212:1215 arg1 gate 1217:1220 a tightly closed pore gate 1195:1220 a tightly closed pore gate 1195:1220 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 6 18 with complex 741:747 arg1 family 784:789 family 784:789 family 784:789 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 18 with complex 741:747 arg1 similarity 805:814 sequence similarity 155 member A (FAM155A) 796:837 sequence similarity 155 member A (FAM155A) 796:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 18 with complex 741:747 arg1 subunit 775:781 a distinct auxiliary subunit 754:781 a distinct auxiliary subunit 754:781 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 3 19 attach linked 325:330 arg1 locomotion 348:357 locomotion 348:357 locomotion 348:357 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 3 19 attach linked 325:330 arg1 respiration 335:345 respiration 335:345 respiration 335:345 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 3 19 attach linked 325:330 arg2 excitability 312:323 neuronal excitability 303:323 neuronal excitability linked to respiration, locomotion and circadian rhythm4-10 303:382 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 3 19 attach linked 325:330 arg1 rhythm4-10 373:382 circadian rhythm4-10 363:382 circadian rhythm4-10 363:382 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 6 20 theme sequence 796:803 arg1 similarity 805:814 sequence similarity 155 member A (FAM155A) 796:837 sequence similarity 155 member A (FAM155A) 796:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 1 21 theme depolarizing 63:74 arg1 currents 94:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents enhance electrical excitability1,2. 32698188 9 22 theme asymmetric 1097:1106 arg1 linkages 1108:1115 asymmetric linkages 1097:1115 asymmetric linkages to the pore 1097:1127 Unusual voltage-sensor domains with asymmetric linkages to the pore suggest mechanisms by which NALCN activity is modulated. 32698188 2 23 from conductance 196:206 arg1 neurons 211:217 neurons 211:217 neurons 211:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 10 24 theme distinctive 1350:1360 arg1 π-bulges 1362:1369 distinctive π-bulges 1350:1369 distinctive π-bulges 1350:1369 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 8 25 theme central 1004:1010 arg1 cavity 1012:1017 a walled-off central cavity 991:1017 a walled-off central cavity with occluded lateral pore fenestrations 991:1058 The pharmacology of NALCN is further delineated by a walled-off central cavity with occluded lateral pore fenestrations. 32698188 1 26 theme sodium 76:81 arg1 currents 94:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents enhance electrical excitability1,2. 32698188 6 27 from structure 719:727 arg1 complex 741:747 complex 741:747 complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A) 741:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 4 28 theme early 494:498 arg1 death15,16 500:509 early death15,16 494:509 early death15,16 494:509 NALCN activity is under tight regulation11-14 and mutations in NALCN cause severe neurological disorders and early death15,16. 32698188 5 29 theme channel 577:583 arg1 assembly 585:592 channel assembly 577:592 channel assembly 577:592 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 2 30 theme ion 143:145 arg1 channel 235:241 the Na+ leak channel 222:241 the Na+ leak channel 222:241 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 30 theme ion 143:145 arg1 channel 147:153 The ion channel 139:153 The ion channel responsible for the major background Na+ conductance in neurons 139:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 9 31 theme Unusual 1061:1067 arg1 domains 1084:1090 Unusual voltage-sensor domains 1061:1090 Unusual voltage-sensor domains with asymmetric linkages to the pore 1061:1127 Unusual voltage-sensor domains with asymmetric linkages to the pore suggest mechanisms by which NALCN activity is modulated. 32698188 4 32 theme NALCN 385:389 arg1 activity 391:398 NALCN activity 385:398 NALCN activity 385:398 NALCN activity is under tight regulation11-14 and mutations in NALCN cause severe neurological disorders and early death15,16. 32698188 1 33 theme Na+ 84:86 arg1 currents 94:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents enhance electrical excitability1,2. 32698188 6 34 dep similarity 805:814 arg1 FAM155A 830:836 FAM155A 830:836 FAM155A 830:836 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 34 dep similarity 805:814 arg1 A 827:827 155 member A 816:827 sequence similarity 155 member A (FAM155A) 796:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 5 35 from channel 531:537 arg1 humans 542:547 humans 542:547 humans 542:547 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 3 36 theme circadian 363:371 arg1 rhythm4-10 373:382 circadian rhythm4-10 363:382 circadian rhythm4-10 363:382 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 5 37 theme assembly 585:592 arg1 selectivity 607:617 ion selectivity 603:617 ion selectivity 603:617 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 37 theme assembly 585:592 arg1 pharmacology 623:634 pharmacology 623:634 pharmacology 623:634 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 37 theme assembly 585:592 arg1 gating 595:600 gating 595:600 gating 595:600 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 37 theme assembly 585:592 arg1 aspects 566:572 fundamental aspects 554:572 fundamental aspects of channel assembly 554:592 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 6 38 theme leak 687:690 arg1 channel 692:698 this essential leak channel 672:698 this essential leak channel 672:698 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 2 39 theme NALCN 259:263 arg1 channel 235:241 the Na+ leak channel 222:241 the Na+ leak channel 222:241 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 39 theme NALCN 259:263 arg1 3,4 265:267 non-selective (NALCN)3,4 244:267 non-selective (NALCN)3,4 244:267 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 40 theme major 175:179 arg1 conductance 196:206 the major background Na+ conductance 171:206 the major background Na+ conductance in neurons 171:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 1 41 theme leak 89:92 arg1 currents 94:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents 50:101 Persistently depolarizing sodium (Na+) leak currents enhance electrical excitability1,2. 32698188 6 42 theme essential 677:685 arg1 channel 692:698 this essential leak channel 672:698 this essential leak channel 672:698 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 11 43 theme treatments 1480:1489 arg1 discovery 1467:1475 discovery 1467:1475 discovery of treatments for NALCN channelopathies and other electrical disorders 1467:1546 Our findings provide a framework to further study the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders. 32698188 0 44 theme leak 30:33 arg1 channel 35:41 the human sodium leak channel 13:41 the human sodium leak channel 13:41 Structure of the human sodium leak channel NALCN. 32698188 9 45 theme voltage-sensor 1069:1082 arg1 domains 1084:1090 Unusual voltage-sensor domains 1061:1090 Unusual voltage-sensor domains with asymmetric linkages to the pore 1061:1127 Unusual voltage-sensor domains with asymmetric linkages to the pore suggest mechanisms by which NALCN activity is modulated. 32698188 2 46 theme Na+ 226:228 arg1 channel 235:241 the Na+ leak channel 222:241 the Na+ leak channel 222:241 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 46 theme Na+ 226:228 arg1 3,4 265:267 non-selective (NALCN)3,4 244:267 non-selective (NALCN)3,4 244:267 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 46 theme Na+ 226:228 arg1 channel 147:153 The ion channel 139:153 The ion channel responsible for the major background Na+ conductance in neurons 139:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 0 47 theme sodium 23:28 arg1 channel 35:41 the human sodium leak channel 13:41 the human sodium leak channel 13:41 Structure of the human sodium leak channel NALCN. 32698188 4 48 theme neurological 467:478 arg1 disorders 480:488 severe neurological disorders 460:488 severe neurological disorders 460:488 NALCN activity is under tight regulation11-14 and mutations in NALCN cause severe neurological disorders and early death15,16. 32698188 0 49 theme human 17:21 arg1 channel 35:41 the human sodium leak channel 13:41 the human sodium leak channel 13:41 Structure of the human sodium leak channel NALCN. 32698188 6 50 theme auxiliary 765:773 arg1 family 784:789 family 784:789 family 784:789 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 50 theme auxiliary 765:773 arg1 subunit 775:781 a distinct auxiliary subunit 754:781 a distinct auxiliary subunit 754:781 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 2 51 theme responsible 155:165 arg1 channel 235:241 the Na+ leak channel 222:241 the Na+ leak channel 222:241 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 51 theme responsible 155:165 arg1 channel 147:153 The ion channel 139:153 The ion channel responsible for the major background Na+ conductance in neurons 139:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 6 52 theme distinct 756:763 arg1 family 784:789 family 784:789 family 784:789 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 52 theme distinct 756:763 arg1 subunit 775:781 a distinct auxiliary subunit 754:781 a distinct auxiliary subunit 754:781 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 11 53 theme electrical 1527:1536 arg1 disorders 1538:1546 other electrical disorders 1521:1546 other electrical disorders 1521:1546 Our findings provide a framework to further study the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders. 32698188 0 54 theme channel 35:41 arg1 Structure 0:8 Structure 0:8 Structure of the human sodium leak channel 0:41 Structure of the human sodium leak channel NALCN. 32698188 4 55 from mutations 435:443 arg1 NALCN 448:452 NALCN 448:452 NALCN 448:452 NALCN activity is under tight regulation11-14 and mutations in NALCN cause severe neurological disorders and early death15,16. 32698188 5 56 theme fundamental 554:564 arg1 selectivity 607:617 ion selectivity 603:617 ion selectivity 603:617 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 56 theme fundamental 554:564 arg1 pharmacology 623:634 pharmacology 623:634 pharmacology 623:634 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 56 theme fundamental 554:564 arg1 gating 595:600 gating 595:600 gating 595:600 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 56 theme fundamental 554:564 arg1 aspects 566:572 fundamental aspects 554:572 fundamental aspects of channel assembly 554:592 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 2 57 theme non-selective 244:256 arg1 channel 235:241 the Na+ leak channel 222:241 the Na+ leak channel 222:241 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 57 theme non-selective 244:256 arg1 3,4 265:267 non-selective (NALCN)3,4 244:267 non-selective (NALCN)3,4 244:267 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 6 58 theme 155 816:818 arg1 FAM155A 830:836 FAM155A 830:836 FAM155A 830:836 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 6 58 theme 155 816:818 arg1 A 827:827 155 member A 816:827 sequence similarity 155 member A (FAM155A) 796:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 1 59 theme electrical 111:120 arg1 excitability1,2 122:136 electrical excitability1,2 111:136 electrical excitability1,2 111:136 Persistently depolarizing sodium (Na+) leak currents enhance electrical excitability1,2. 32698188 11 60 theme other 1521:1525 arg1 disorders 1538:1546 other electrical disorders 1521:1546 other electrical disorders 1521:1546 Our findings provide a framework to further study the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders. 32698188 10 61 theme patient 1262:1268 arg1 mutations 1270:1278 missense patient mutations 1253:1278 missense patient mutations 1253:1278 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 3 62 theme NALCN-mediated 270:283 arg1 currents 285:292 NALCN-mediated currents 270:292 NALCN-mediated currents 270:292 NALCN-mediated currents regulate neuronal excitability linked to respiration, locomotion and circadian rhythm4-10. 32698188 2 63 theme Na+ 192:194 arg1 conductance 196:206 the major background Na+ conductance 171:206 the major background Na+ conductance in neurons 171:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 10 64 theme gain-of-function 1286:1301 arg1 phenotypes 1303:1312 gain-of-function phenotypes 1286:1312 gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges 1286:1369 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 5 65 theme orphan 524:529 arg1 NALCN 512:516 NALCN 512:516 NALCN 512:516 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 5 65 theme orphan 524:529 arg1 channel 531:537 an orphan channel 521:537 an orphan channel in humans 521:547 NALCN is an orphan channel in humans, and fundamental aspects of channel assembly, gating, ion selectivity and pharmacology remain obscure. 32698188 10 66 theme S6 1338:1339 arg1 gate 1341:1344 the S6 gate 1334:1344 the S6 gate 1334:1344 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 11 67 theme NALCN 1495:1499 arg1 channelopathies 1501:1515 NALCN channelopathies 1495:1515 NALCN channelopathies 1495:1515 Our findings provide a framework to further study the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders. 32698188 2 68 theme background 181:190 arg1 conductance 196:206 the major background Na+ conductance 171:206 the major background Na+ conductance in neurons 171:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 69 theme leak 230:233 arg1 channel 235:241 the Na+ leak channel 222:241 the Na+ leak channel 222:241 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 69 theme leak 230:233 arg1 3,4 265:267 non-selective (NALCN)3,4 244:267 non-selective (NALCN)3,4 244:267 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 2 69 theme leak 230:233 arg1 channel 147:153 The ion channel 139:153 The ion channel responsible for the major background Na+ conductance in neurons 139:217 The ion channel responsible for the major background Na+ conductance in neurons is the Na+ leak channel, non-selective (NALCN)3,4. 32698188 6 70 theme NALCN 732:736 arg1 structure 719:727 the structure 715:727 the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A) 715:837 Here we investigate this essential leak channel and determined the structure of NALCN in complex with a distinct auxiliary subunit, family with sequence similarity 155 member A (FAM155A). 32698188 10 71 theme closed 1205:1210 arg1 gate 1217:1220 a tightly closed pore gate 1195:1220 a tightly closed pore gate 1195:1220 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 10 72 theme mutations 1270:1278 arg1 majority 1241:1248 the majority 1237:1248 the majority of missense patient mutations 1237:1278 We found a tightly closed pore gate in NALCN where the majority of missense patient mutations cause gain-of-function phenotypes that cluster around the S6 gate and distinctive π-bulges. 32698188 11 73 theme NALCN 1440:1444 arg1 physiology 1426:1435 the physiology 1422:1435 the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders 1422:1546 Our findings provide a framework to further study the physiology of NALCN and a foundation for discovery of treatments for NALCN channelopathies and other electrical disorders. 9233787 5 0 theme opposite 683:690 arg1 BM-40 676:680 BM-40 676:680 BM-40 676:680 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 6 1 theme Kazal 842:846 arg1 family 848:853 the Kazal family 838:853 the Kazal family 838:853 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 3 2 from resolution 391:400 arg1 structure 372:380 the crystal structure 360:380 the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40 360:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 7 3 theme striking 973:980 arg1 similarities 982:993 striking similarities 973:993 striking similarities to epidermal growth factor 973:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 4 4 theme EF-hand 521:527 arg1 pair 529:532 the EF-hand pair 517:532 the EF-hand pair of the EC domain 517:549 The two distinct domains interact through a small interface that involves the EF-hand pair of the EC domain. 9233787 0 5 theme domains 76:82 arg1 pair 23:26 a pair 21:26 a pair of follistatin-like and EF-hand calcium-binding domains in BM-40 21:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 4 6 theme distinct 451:458 arg1 domains 460:466 The two distinct domains 443:466 The two distinct domains 443:466 The two distinct domains interact through a small interface that involves the EF-hand pair of the EC domain. 9233787 2 7 theme calcium-binding 316:330 arg1 domain 337:342 an EF-hand calcium-binding (EC) domain 305:342 an EF-hand calcium-binding (EC) domain 305:342 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 8 8 theme rigid 1058:1062 arg1 spacer 1064:1069 a rigid spacer 1056:1069 a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin 1056:1152 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 8 theme rigid 1058:1062 arg1 hairpin 1028:1034 This hairpin 1023:1034 This hairpin 1023:1034 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 8 theme rigid 1058:1062 arg1 site 1185:1188 the heparin-binding site 1165:1188 the heparin-binding site in follistatin 1165:1203 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 1 9 gly glycoprotein 166:177 arg1 glycoprotein 166:177 an anti-adhesive secreted glycoprotein 140:177 an anti-adhesive secreted glycoprotein involved in tissue remodelling 140:208 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 1 9 gly glycoprotein 166:177 arg1 BM-40 94:98 BM-40 94:98 BM-40 (also known as SPARC or osteonectin) 94:135 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 8 10 theme repeated 1103:1110 arg1 domains 1115:1121 tandemly repeated FS domains 1094:1121 tandemly repeated FS domains 1094:1121 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 3 11 theme human 430:434 arg1 BM-40 436:440 human BM-40 430:440 human BM-40 430:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 8 12 theme heparin-binding 1169:1183 arg1 spacer 1064:1069 a rigid spacer 1056:1069 a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin 1056:1152 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 12 theme heparin-binding 1169:1183 arg1 site 1185:1188 the heparin-binding site 1165:1188 the heparin-binding site in follistatin 1165:1203 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 12 theme heparin-binding 1169:1183 arg1 hairpin 1028:1034 This hairpin 1023:1034 This hairpin 1023:1034 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 7 13 with beta-hairpin 955:966 arg1 similarities 982:993 striking similarities 973:993 striking similarities to epidermal growth factor 973:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 6 14 theme serine 808:813 arg1 inhibitors 824:833 serine protease inhibitors 808:833 serine protease inhibitors of the Kazal family 808:853 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 5 15 theme binding 696:702 arg1 epitope 704:710 the binding epitope 692:710 the binding epitope for collagens and the N-linked carbohydrate 692:754 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 1 16 dep known 106:110 arg1 also 101:104 also 101:104 also 101:104 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 1 17 theme anti-adhesive 143:155 arg1 glycoprotein 166:177 an anti-adhesive secreted glycoprotein 140:177 an anti-adhesive secreted glycoprotein involved in tissue remodelling 140:208 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 1 17 theme anti-adhesive 143:155 arg1 BM-40 94:98 BM-40 94:98 BM-40 (also known as SPARC or osteonectin) 94:135 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 0 18 from pair 23:26 arg1 BM-40 87:91 BM-40 87:91 BM-40 87:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 5 19 theme BM-40 676:680 arg1 face 668:671 one face 664:671 one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate 664:754 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 6 20 theme protease 815:822 arg1 inhibitors 824:833 serine protease inhibitors 808:833 serine protease inhibitors of the Kazal family 808:853 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 1 21 theme secreted 157:164 arg1 glycoprotein 166:177 an anti-adhesive secreted glycoprotein 140:177 an anti-adhesive secreted glycoprotein involved in tissue remodelling 140:208 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 1 21 theme secreted 157:164 arg1 BM-40 94:98 BM-40 94:98 BM-40 (also known as SPARC or osteonectin) 94:135 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 0 22 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 0:92 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 5 23 theme cell 575:578 arg1 binding 580:586 cell binding 575:586 cell binding 575:586 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 8 24 from spacer 1064:1069 arg1 follistatin 1132:1142 follistatin 1132:1142 follistatin 1132:1142 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 24 from spacer 1064:1069 arg1 proteins 1074:1081 proteins 1074:1081 proteins 1074:1081 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 24 from spacer 1064:1069 arg1 agrin 1148:1152 agrin 1148:1152 agrin 1148:1152 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 5 25 theme N-linked 734:741 arg1 carbohydrate 743:754 the N-linked carbohydrate 730:754 the N-linked carbohydrate 730:754 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 4 26 theme small 487:491 arg1 interface 493:501 a small interface 485:501 a small interface that involves the EF-hand pair of the EC domain 485:549 The two distinct domains interact through a small interface that involves the EF-hand pair of the EC domain. 9233787 5 27 dep opposite 683:690 arg1 epitope 704:710 the binding epitope 692:710 the binding epitope for collagens and the N-linked carbohydrate 692:754 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 7 28 theme Notable 856:862 arg1 differences 864:874 Notable differences 856:874 Notable differences 856:874 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 7 28 theme Notable 856:862 arg1 insertion 883:891 an insertion 880:891 an insertion into the inhibitory loop in BM-40 880:925 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 3 29 theme FS-EC 409:413 arg1 pair 422:425 the FS-EC domain pair 405:425 the FS-EC domain pair of human BM-40 405:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 8 30 dep likely 1039:1044 arg1 forms 1159:1163 forms 1159:1163 forms the heparin-binding site in follistatin 1159:1203 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 30 dep likely 1039:1044 arg1 act 1049:1051 act 1049:1051 to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin 1046:1152 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 5 31 theme focal 637:641 arg1 adhesions 643:651 focal adhesions 637:651 focal adhesions 637:651 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 2 32 theme follistatin-like 272:287 arg1 domain 294:299 a follistatin-like (FS) domain 270:299 a follistatin-like (FS) domain 270:299 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 2 32 theme follistatin-like 272:287 arg1 FS 290:291 FS 290:291 FS 290:291 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 3 33 theme domain 415:420 arg1 pair 422:425 the FS-EC domain pair 405:425 the FS-EC domain pair of human BM-40 405:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 3 34 theme crystal 364:370 arg1 structure 372:380 the crystal structure 360:380 the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40 360:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 5 35 theme adhesions 643:651 arg1 disassembly 622:632 disassembly 622:632 disassembly of focal adhesions 622:651 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 5 35 theme adhesions 643:651 arg1 inhibition 589:598 inhibition 589:598 inhibition of cell spreading 589:616 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 5 35 theme adhesions 643:651 arg1 binding 580:586 cell binding 575:586 cell binding 575:586 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 0 36 from domains 76:82 arg1 BM-40 87:91 BM-40 87:91 BM-40 87:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 1 37 theme tissue 191:196 arg1 remodelling 198:208 tissue remodelling 191:208 tissue remodelling 191:208 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 0 38 theme pair 23:26 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 0:92 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 7 39 theme epidermal 998:1006 arg1 factor 1015:1020 epidermal growth factor 998:1020 epidermal growth factor 998:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 7 40 theme protruding 933:942 arg1 beta-hairpin 955:966 a protruding N-terminal beta-hairpin 931:966 a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor 931:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 7 41 theme growth 1008:1013 arg1 factor 1015:1020 epidermal growth factor 998:1020 epidermal growth factor 998:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 5 42 theme cell 603:606 arg1 spreading 608:616 cell spreading 603:616 cell spreading 603:616 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 7 43 theme N-terminal 944:953 arg1 beta-hairpin 955:966 a protruding N-terminal beta-hairpin 931:966 a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor 931:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 2 44 theme EF-hand 308:314 arg1 domain 337:342 an EF-hand calcium-binding (EC) domain 305:342 an EF-hand calcium-binding (EC) domain 305:342 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 0 45 theme follistatin-like 31:46 arg1 domains 76:82 follistatin-like and EF-hand calcium-binding domains 31:82 follistatin-like and EF-hand calcium-binding domains in BM-40 31:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 5 46 theme spreading 608:616 arg1 disassembly 622:632 disassembly 622:632 disassembly of focal adhesions 622:651 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 5 46 theme spreading 608:616 arg1 inhibition 589:598 inhibition 589:598 inhibition of cell spreading 589:616 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 5 46 theme spreading 608:616 arg1 binding 580:586 cell binding 575:586 cell binding 575:586 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 6 47 theme FS 771:772 arg1 domain 774:779 The elongated FS domain 757:779 The elongated FS domain 757:779 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 6 47 theme FS 771:772 arg1 related 797:803 related 797:803 related 797:803 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 3 48 theme BM-40 436:440 arg1 pair 422:425 the FS-EC domain pair 405:425 the FS-EC domain pair of human BM-40 405:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 6 49 theme elongated 761:769 arg1 domain 774:779 The elongated FS domain 757:779 The elongated FS domain 757:779 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 6 49 theme elongated 761:769 arg1 related 797:803 related 797:803 related 797:803 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 5 50 link N-linked 734:741 arg1 carbohydrate 743:754 the N-linked carbohydrate 730:754 the N-linked carbohydrate 730:754 Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM-40, opposite the binding epitope for collagens and the N-linked carbohydrate. 9233787 3 51 theme pair 422:425 arg1 resolution 391:400 3.1 A resolution 385:400 3.1 A resolution of the FS-EC domain pair of human BM-40 385:440 Here we report the crystal structure at 3.1 A resolution of the FS-EC domain pair of human BM-40. 9233787 8 52 from site 1185:1188 arg1 follistatin 1193:1203 follistatin 1193:1203 follistatin 1193:1203 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 4 53 theme EC 541:542 arg1 domain 544:549 the EC domain 537:549 the EC domain 537:549 The two distinct domains interact through a small interface that involves the EF-hand pair of the EC domain. 9233787 8 54 contain containing 1083:1092 arg1 follistatin 1132:1142 follistatin 1132:1142 follistatin 1132:1142 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 54 contain containing 1083:1092 arg2 domains 1115:1121 tandemly repeated FS domains 1094:1121 tandemly repeated FS domains 1094:1121 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 54 contain containing 1083:1092 arg1 proteins 1074:1081 proteins 1074:1081 proteins 1074:1081 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 54 contain containing 1083:1092 arg1 agrin 1148:1152 agrin 1148:1152 agrin 1148:1152 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 8 55 theme FS 1112:1113 arg1 domains 1115:1121 tandemly repeated FS domains 1094:1121 tandemly repeated FS domains 1094:1121 This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin-binding site in follistatin. 9233787 0 56 theme calcium-binding 60:74 arg1 domains 76:82 follistatin-like and EF-hand calcium-binding domains 31:82 follistatin-like and EF-hand calcium-binding domains in BM-40 31:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 0 57 from BM-40 87:91 arg1 pair 23:26 a pair 21:26 a pair of follistatin-like and EF-hand calcium-binding domains in BM-40 21:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 2 58 theme N-terminal 232:241 arg1 segment 243:249 an acidic N-terminal segment 222:249 an acidic N-terminal segment 222:249 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 1 59 dep BM-40 94:98 arg1 known 106:110 known 106:110 known as SPARC or osteonectin 106:134 BM-40 (also known as SPARC or osteonectin) is an anti-adhesive secreted glycoprotein involved in tissue remodelling. 9233787 7 60 from loop 913:916 arg1 BM-40 921:925 BM-40 921:925 BM-40 921:925 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 0 61 theme EF-hand 52:58 arg1 domains 76:82 follistatin-like and EF-hand calcium-binding domains 31:82 follistatin-like and EF-hand calcium-binding domains in BM-40 31:91 Crystal structure of a pair of follistatin-like and EF-hand calcium-binding domains in BM-40. 9233787 6 62 theme family 848:853 arg1 inhibitors 824:833 serine protease inhibitors 808:833 serine protease inhibitors of the Kazal family 808:853 The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. 9233787 2 63 theme acidic 225:230 arg1 segment 243:249 an acidic N-terminal segment 222:249 an acidic N-terminal segment 222:249 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 7 64 theme inhibitory 902:911 arg1 loop 913:916 the inhibitory loop 898:916 the inhibitory loop in BM-40 898:925 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 2 65 theme EC 333:334 arg1 domain 337:342 an EF-hand calcium-binding (EC) domain 305:342 an EF-hand calcium-binding (EC) domain 305:342 Apart from an acidic N-terminal segment, BM-40 consists of a follistatin-like (FS) domain and an EF-hand calcium-binding (EC) domain. 9233787 7 66 with insertion 883:891 arg1 similarities 982:993 striking similarities 973:993 striking similarities to epidermal growth factor 973:1020 Notable differences are an insertion into the inhibitory loop in BM-40 and a protruding N-terminal beta-hairpin with striking similarities to epidermal growth factor. 9233787 4 67 theme domain 544:549 arg1 pair 529:532 the EF-hand pair 517:532 the EF-hand pair of the EC domain 517:549 The two distinct domains interact through a small interface that involves the EF-hand pair of the EC domain. 4054110 0 0 theme kininogen 72:80 arg1 chain 37:41 the light chain 27:41 the light chain of human high-molecular-mass kininogen 27:80 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 6 1 theme amino 599:603 arg1 acids 605:609 22 amino acids 596:609 22 amino acids within the histidine-rich part of the sequence 596:656 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 0 2 theme high-molecular-mass 52:70 arg1 kininogen 72:80 human high-molecular-mass kininogen 46:80 human high-molecular-mass kininogen 46:80 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 1 3 theme human 138:142 arg1 kininogen 164:172 human high-molecular-mass kininogen 138:172 human high-molecular-mass kininogen 138:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 5 4 theme carbohydrate 401:412 arg1 chains 419:424 All carbohydrate side chains 397:424 All carbohydrate side chains 397:424 All carbohydrate side chains are O-glycosidically linked. 4054110 7 5 theme gene 704:707 arg1 multiplication 709:722 gene multiplication 704:722 gene multiplication 704:722 The histidine-rich region may have arisen by gene multiplication during evolution. 4054110 6 6 theme chain 521:525 arg1 sequence 527:534 the bovine kininogen light chain sequence 494:534 the bovine kininogen light chain sequence 494:534 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 4 7 theme attachment 368:377 arg1 sites 379:383 Nine carbohydrate attachment sites 350:383 Nine carbohydrate attachment sites 350:383 Nine carbohydrate attachment sites were found. 4054110 5 8 theme side 414:417 arg1 chains 419:424 All carbohydrate side chains 397:424 All carbohydrate side chains 397:424 All carbohydrate side chains are O-glycosidically linked. 4054110 6 9 theme light 515:519 arg1 chain 521:525 the bovine kininogen light chain 494:525 the bovine kininogen light chain sequence 494:534 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 6 10 theme kininogen 505:513 arg1 chain 521:525 the bovine kininogen light chain 494:525 the bovine kininogen light chain sequence 494:534 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 1 11 theme high-molecular-mass 144:162 arg1 kininogen 164:172 human high-molecular-mass kininogen 138:172 human high-molecular-mass kininogen 138:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 6 12 theme bovine 498:503 arg1 chain 521:525 the bovine kininogen light chain 494:525 the bovine kininogen light chain sequence 494:534 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 1 13 theme kininogen 164:172 arg1 chain 129:133 the light chain 119:133 the light chain of human high-molecular-mass kininogen 119:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 0 14 theme amino 4:8 arg1 sequence 15:22 The amino acid sequence 0:22 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 0:81 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 3 15 theme heavy 305:309 arg1 chain 311:315 the heavy chain 301:315 the heavy chain 301:315 The half-cystine, which forms the disulfide bridge to the heavy chain, was identified in position 225. 4054110 6 16 theme high 544:547 arg1 degree 549:554 a high degree 542:554 a high degree of homology 542:566 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 4 17 theme carbohydrate 355:366 arg1 sites 379:383 Nine carbohydrate attachment sites 350:383 Nine carbohydrate attachment sites 350:383 Nine carbohydrate attachment sites were found. 4054110 6 18 with Alignment 455:463 arg1 sequence 527:534 the bovine kininogen light chain sequence 494:534 the bovine kininogen light chain sequence 494:534 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 1 19 theme complete 87:94 arg1 sequence 107:114 The complete amino acid sequence 83:114 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen 83:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 6 20 theme sequence 649:656 arg1 part 637:640 the histidine-rich part 618:640 the histidine-rich part of the sequence 618:656 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 0 21 theme acid 10:13 arg1 sequence 15:22 The amino acid sequence 0:22 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 0:81 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 1 22 theme amino 96:100 arg1 sequence 107:114 The complete amino acid sequence 83:114 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen 83:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 6 23 theme sequence 480:487 arg1 Alignment 455:463 Alignment 455:463 Alignment of the present sequence with the bovine kininogen light chain sequence 455:534 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 2 24 theme amino 226:230 arg1 residues 237:244 255 amino acid residues 222:244 255 amino acid residues 222:244 The peptide chain contains 255 amino acid residues. 4054110 6 25 theme homology 559:566 arg1 degree 549:554 a high degree 542:554 a high degree of homology 542:566 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 6 26 theme present 472:478 arg1 sequence 480:487 the present sequence 468:487 the present sequence 468:487 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 0 27 theme chain 37:41 arg1 sequence 15:22 The amino acid sequence 0:22 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 0:81 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 7 28 theme histidine-rich 663:676 arg1 region 678:683 The histidine-rich region 659:683 The histidine-rich region 659:683 The histidine-rich region may have arisen by gene multiplication during evolution. 4054110 1 29 theme acid 102:105 arg1 sequence 107:114 The complete amino acid sequence 83:114 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen 83:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 0 30 theme light 31:35 arg1 chain 37:41 the light chain 27:41 the light chain of human high-molecular-mass kininogen 27:80 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 2 31 theme peptide 199:205 arg1 chain 207:211 The peptide chain 195:211 The peptide chain 195:211 The peptide chain contains 255 amino acid residues. 4054110 6 32 theme histidine-rich 622:635 arg1 part 637:640 the histidine-rich part 618:640 the histidine-rich part of the sequence 618:656 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 4054110 3 33 theme disulfide 281:289 arg1 bridge 291:296 the disulfide bridge 277:296 the disulfide bridge to the heavy chain 277:315 The half-cystine, which forms the disulfide bridge to the heavy chain, was identified in position 225. 4054110 2 34 contain contains 213:220 arg1 chain 207:211 The peptide chain 195:211 The peptide chain 195:211 The peptide chain contains 255 amino acid residues. 4054110 2 34 contain contains 213:220 arg2 residues 237:244 255 amino acid residues 222:244 255 amino acid residues 222:244 The peptide chain contains 255 amino acid residues. 4054110 0 35 theme human 46:50 arg1 kininogen 72:80 human high-molecular-mass kininogen 46:80 human high-molecular-mass kininogen 46:80 The amino acid sequence of the light chain of human high-molecular-mass kininogen. 4054110 1 36 theme light 123:127 arg1 chain 129:133 the light chain 119:133 the light chain of human high-molecular-mass kininogen 119:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 2 37 theme acid 232:235 arg1 residues 237:244 255 amino acid residues 222:244 255 amino acid residues 222:244 The peptide chain contains 255 amino acid residues. 4054110 1 38 theme chain 129:133 arg1 sequence 107:114 The complete amino acid sequence 83:114 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen 83:172 The complete amino acid sequence of the light chain of human high-molecular-mass kininogen has been determined. 4054110 6 39 theme acids 605:609 arg1 extension 583:591 an extension 580:591 an extension of 22 amino acids within the histidine-rich part of the sequence 580:656 Alignment of the present sequence with the bovine kininogen light chain sequence shows a high degree of homology, except for an extension of 22 amino acids within the histidine-rich part of the sequence. 2991887 5 0 theme preproleader 525:536 arg1 sequence 538:545 a preproleader sequence 523:545 a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids 523:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 1 1 theme protein 115:121 arg1 C 123:123 protein C 115:123 protein C 115:123 A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. 2991887 9 2 theme factor 1194:1199 arg1 IX 1201:1202 human factor IX 1188:1202 human factor IX 1188:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 5 3 theme Lys-Arg 626:632 arg1 chain 574:578 a light chain 566:578 a light chain of 155 amino acids 566:597 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 3 theme Lys-Arg 626:632 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 3 theme Lys-Arg 626:632 arg1 chain 647:651 a heavy chain 639:651 a heavy chain of 262 amino acids 639:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 3 theme Lys-Arg 626:632 arg1 dipeptide 613:621 a connecting dipeptide 600:621 a connecting dipeptide of Lys-Arg 600:632 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 9 4 theme first 1215:1219 arg1 located 1252:1258 located 1252:1258 located 1252:1258 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 4 theme first 1215:1219 arg1 introns 1227:1233 the first three introns 1211:1233 the first three introns in protein C 1211:1246 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 5 from positions 1272:1280 arg1 located 1252:1258 located 1252:1258 located 1252:1258 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 5 from positions 1272:1280 arg1 introns 1227:1233 the first three introns 1211:1233 the first three introns in protein C 1211:1246 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 6 theme human 1188:1192 arg1 IX 1201:1202 human factor IX 1188:1202 human factor IX 1188:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 5 7 theme amino 587:591 arg1 acids 593:597 155 amino acids 583:597 155 amino acids 583:597 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 7 8 theme serine 923:928 arg1 protease 930:937 the serine protease 919:937 the serine protease 919:937 The heavy chain also contains the catalytic region for the serine protease. 2991887 6 9 theme light 817:821 arg1 chain 823:827 a light chain 815:827 a light chain 815:827 The preproleader sequence and the connecting dipeptide are removed during processing, resulting in the mature protein composed of a heavy and a light chain held together by a disulfide bond. 2991887 3 10 theme complete 295:302 arg1 sequence 304:311 The complete sequence 291:311 The complete sequence of the gene 291:323 The complete sequence of the gene was determined by the dideoxy method and shown to span about 11 kilobases of DNA. 2991887 9 11 located located 1082:1088 arg2 introns 1044:1050 The seven introns 1034:1050 The seven introns in the gene for protein C 1034:1076 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 11 located located 1082:1088 arg2 located 1082:1088 located 1082:1088 located 1082:1088 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 11 located located 1082:1088 arg1 positions 1114:1122 essentially the same positions 1093:1122 essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX 1093:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 12 from located 1082:1088 arg1 positions 1114:1122 essentially the same positions 1093:1122 essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX 1093:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 2 13 theme protein 280:286 arg1 C 288:288 protein C 280:288 protein C 280:288 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 9 14 theme same 1109:1112 arg1 positions 1114:1122 essentially the same positions 1093:1122 essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX 1093:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 8 15 theme intron 1024:1029 arg1 E 1031:1031 intron E 1024:1031 intron E 1024:1031 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 9 16 from located 1252:1258 arg1 positions 1272:1280 the same positions 1263:1280 the same positions as the first three in the gene for human prothrombin 1263:1333 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 2 17 theme 4A 219:220 arg1 phage 222:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 2 18 dep different 183:191 arg1 overlapping 193:203 overlapping 193:203 overlapping 193:203 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 0 19 theme nucleotide 4:13 arg1 sequence 15:22 The nucleotide sequence 0:22 The nucleotide sequence of the gene for human protein C 0:54 The nucleotide sequence of the gene for human protein C. 2991887 7 20 theme catalytic 898:906 arg1 region 908:913 the catalytic region 894:913 the catalytic region for the serine protease 894:937 The heavy chain also contains the catalytic region for the serine protease. 2991887 1 21 theme human 59:63 arg1 library 77:83 A human genomic DNA library 57:83 A human genomic DNA library 57:83 A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. 2991887 7 22 theme heavy 868:872 arg1 chain 874:878 The heavy chain 864:878 The heavy chain 864:878 The heavy chain also contains the catalytic region for the serine protease. 2991887 3 23 theme dideoxy 347:353 arg1 method 355:360 the dideoxy method 343:360 the dideoxy method 343:360 The complete sequence of the gene was determined by the dideoxy method and shown to span about 11 kilobases of DNA. 2991887 3 24 theme gene 320:323 arg1 sequence 304:311 The complete sequence 291:311 The complete sequence of the gene 291:323 The complete sequence of the gene was determined by the dideoxy method and shown to span about 11 kilobases of DNA. 2991887 4 25 theme gene 450:453 arg1 portion 435:441 The coding and 3' noncoding portion 407:441 The coding and 3' noncoding portion of the gene 407:453 The coding and 3' noncoding portion of the gene consists of eight exons and seven introns. 2991887 9 26 theme protein 1068:1074 arg1 C 1076:1076 protein C 1068:1076 protein C 1068:1076 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 1 27 theme genomic 65:71 arg1 library 77:83 A human genomic DNA library 57:83 A human genomic DNA library 57:83 A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. 2991887 1 28 theme cDNA 136:139 arg1 probe 141:145 a cDNA probe 134:145 a cDNA probe coding for the human protein 134:174 A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. 2991887 5 29 theme light 568:572 arg1 chain 574:578 a light chain 566:578 a light chain of 155 amino acids 566:597 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 29 theme light 568:572 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 1 30 theme DNA 73:75 arg1 library 77:83 A human genomic DNA library 57:83 A human genomic DNA library 57:83 A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. 2991887 5 31 theme acids 666:670 arg1 chain 574:578 a light chain 566:578 a light chain of 155 amino acids 566:597 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 31 theme acids 666:670 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 31 theme acids 666:670 arg1 chain 647:651 a heavy chain 639:651 a heavy chain of 262 amino acids 639:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 31 theme acids 666:670 arg1 dipeptide 613:621 a connecting dipeptide 600:621 a connecting dipeptide of Lys-Arg 600:632 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 32 theme amino 660:664 arg1 acids 666:670 262 amino acids 656:670 262 amino acids 656:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 9 33 theme amino 1131:1135 arg1 sequence 1142:1149 the amino acid sequence 1127:1149 the amino acid sequence as the seven introns in the gene for human factor IX 1127:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 6 34 theme connecting 707:716 arg1 dipeptide 718:726 the connecting dipeptide 703:726 the connecting dipeptide 703:726 The preproleader sequence and the connecting dipeptide are removed during processing, resulting in the mature protein composed of a heavy and a light chain held together by a disulfide bond. 2991887 9 35 theme human 1317:1321 arg1 prothrombin 1323:1333 human prothrombin 1317:1333 human prothrombin 1317:1333 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 0 36 theme gene 31:34 arg1 sequence 15:22 The nucleotide sequence 0:22 The nucleotide sequence of the gene for human protein C 0:54 The nucleotide sequence of the gene for human protein C. 2991887 9 37 theme acid 1137:1140 arg1 sequence 1142:1149 the amino acid sequence 1127:1149 the amino acid sequence as the seven introns in the gene for human factor IX 1127:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 38 from introns 1044:1050 arg1 gene 1059:1062 the gene 1055:1062 the gene for protein C 1055:1076 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 7 39 contain contains 885:892 arg2 region 908:913 the catalytic region 894:913 the catalytic region for the serine protease 894:937 The heavy chain also contains the catalytic region for the serine protease. 2991887 7 39 contain contains 885:892 arg1 chain 874:878 The heavy chain 864:878 The heavy chain 864:878 The heavy chain also contains the catalytic region for the serine protease. 2991887 9 40 from positions 1114:1122 arg1 located 1082:1088 located 1082:1088 located 1082:1088 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 40 from positions 1114:1122 arg1 introns 1044:1050 The seven introns 1034:1050 The seven introns in the gene for protein C 1034:1076 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 40 from positions 1114:1122 arg1 sequence 1142:1149 the amino acid sequence 1127:1149 the amino acid sequence as the seven introns in the gene for human factor IX 1127:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 4 41 theme noncoding 425:433 arg1 portion 435:441 The coding and 3' noncoding portion 407:441 The coding and 3' noncoding portion of the gene 407:453 The coding and 3' noncoding portion of the gene consists of eight exons and seven introns. 2991887 9 42 located located 1252:1258 arg2 introns 1227:1233 the first three introns 1211:1233 the first three introns in protein C 1211:1246 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 42 located located 1252:1258 arg1 positions 1272:1280 the same positions 1263:1280 the same positions as the first three in the gene for human prothrombin 1263:1333 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 9 42 located located 1252:1258 arg2 located 1252:1258 located 1252:1258 located 1252:1258 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 8 43 theme nucleotides 998:1008 arg1 repeats 977:983 two homologous repeats 962:983 two homologous repeats 962:983 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 8 43 theme nucleotides 998:1008 arg1 sequences 948:956 Two Alu sequences 940:956 Two Alu sequences 940:956 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 0 44 theme human 40:44 arg1 C 54:54 human protein C 40:54 human protein C 40:54 The nucleotide sequence of the gene for human protein C. 2991887 2 45 contain contain 247:253 arg2 inserts 255:261 inserts 255:261 inserts for the gene for protein C 255:288 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 2 45 contain contain 247:253 arg1 that 242:245 that 242:245 that 242:245 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 5 46 theme amino 553:557 arg1 chain 574:578 a light chain 566:578 a light chain of 155 amino acids 566:597 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 46 theme amino 553:557 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 46 theme amino 553:557 arg1 chain 647:651 a heavy chain 639:651 a heavy chain of 262 amino acids 639:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 46 theme amino 553:557 arg1 dipeptide 613:621 a connecting dipeptide 600:621 a connecting dipeptide of Lys-Arg 600:632 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 8 47 theme Alu 944:946 arg1 sequences 948:956 Two Alu sequences 940:956 Two Alu sequences 940:956 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 3 48 theme DNA 402:404 arg1 kilobases 389:397 about 11 kilobases 380:397 about 11 kilobases of DNA 380:404 The complete sequence of the gene was determined by the dideoxy method and shown to span about 11 kilobases of DNA. 2991887 5 49 theme heavy 641:645 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 49 theme heavy 641:645 arg1 chain 647:651 a heavy chain 639:651 a heavy chain of 262 amino acids 639:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 6 50 theme mature 776:781 arg1 protein 783:789 the mature protein 772:789 the mature protein composed of a heavy and a light chain held together by a disulfide bond 772:861 The preproleader sequence and the connecting dipeptide are removed during processing, resulting in the mature protein composed of a heavy and a light chain held together by a disulfide bond. 2991887 5 51 theme acids 559:563 arg1 sequence 538:545 a preproleader sequence 523:545 a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids 523:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 6 52 theme preproleader 677:688 arg1 sequence 690:697 The preproleader sequence 673:697 The preproleader sequence 673:697 The preproleader sequence and the connecting dipeptide are removed during processing, resulting in the mature protein composed of a heavy and a light chain held together by a disulfide bond. 2991887 8 53 located found 1015:1019 arg2 repeats 977:983 two homologous repeats 962:983 two homologous repeats 962:983 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 8 53 located found 1015:1019 arg1 E 1031:1031 intron E 1024:1031 intron E 1024:1031 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 8 53 located found 1015:1019 arg2 sequences 948:956 Two Alu sequences 940:956 Two Alu sequences 940:956 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 9 54 from introns 1164:1170 arg1 gene 1179:1182 the gene 1175:1182 the gene for human factor IX 1175:1202 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 8 55 theme homologous 966:975 arg1 repeats 977:983 two homologous repeats 962:983 two homologous repeats 962:983 Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. 2991887 2 56 theme Charon 212:217 arg1 phage 222:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 9 57 from introns 1227:1233 arg1 C 1246:1246 protein C 1238:1246 protein C 1238:1246 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 1 58 theme human 162:166 arg1 protein 168:174 the human protein 158:174 the human protein 158:174 A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. 2991887 2 59 theme lambda 205:210 arg1 phage 222:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 9 60 theme same 1267:1270 arg1 positions 1272:1280 the same positions 1263:1280 the same positions as the first three in the gene for human prothrombin 1263:1333 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 5 61 theme connecting 602:611 arg1 dipeptide 613:621 a connecting dipeptide 600:621 a connecting dipeptide of Lys-Arg 600:632 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 61 theme connecting 602:611 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 9 62 theme protein 1238:1244 arg1 C 1246:1246 protein C 1238:1246 protein C 1238:1246 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 0 63 theme protein 46:52 arg1 C 54:54 human protein C 40:54 human protein C 40:54 The nucleotide sequence of the gene for human protein C. 2991887 4 64 theme coding 411:416 arg1 portion 435:441 The coding and 3' noncoding portion 407:441 The coding and 3' noncoding portion of the gene 407:453 The coding and 3' noncoding portion of the gene consists of eight exons and seven introns. 2991887 2 65 theme different 183:191 arg1 phage 222:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage 177:226 Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. 2991887 6 66 theme disulfide 848:856 arg1 bond 858:861 a disulfide bond 846:861 a disulfide bond 846:861 The preproleader sequence and the connecting dipeptide are removed during processing, resulting in the mature protein composed of a heavy and a light chain held together by a disulfide bond. 2991887 9 67 from three 1295:1299 arg1 gene 1308:1311 the gene 1304:1311 the gene for human prothrombin 1304:1333 The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. 2991887 5 68 theme acids 593:597 arg1 chain 574:578 a light chain 566:578 a light chain of 155 amino acids 566:597 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 68 theme acids 593:597 arg1 acids 559:563 42 amino acids 550:563 42 amino acids 550:563 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 68 theme acids 593:597 arg1 chain 647:651 a heavy chain 639:651 a heavy chain of 262 amino acids 639:670 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 2991887 5 68 theme acids 593:597 arg1 dipeptide 613:621 a connecting dipeptide 600:621 a connecting dipeptide of Lys-Arg 600:632 The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. 19329486 5 0 theme responsible 614:624 arg1 protein 605:611 an MC2R accessory protein 587:611 an MC2R accessory protein 587:611 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 5 0 theme responsible 614:624 arg1 MRAP 579:582 MRAP 579:582 MRAP 579:582 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 9 1 dep hormone 1018:1024 arg1 D-Phe7 982:987 [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone 976:1024 D-Phe7 982:987 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 9 2 theme MC5R 953:956 arg1 MC1R 931:934 MC1R 931:934 MC1R 931:934 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 9 2 theme MC5R 953:956 arg1 responsiveness 958:971 MC5R responsiveness 953:971 MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH) 953:1034 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 0 3 theme family 73:78 arg1 MRAP2 9:13 MRAP2 9:13 MRAP2 9:13 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 0 3 theme family 73:78 arg1 regulators 33:42 bidirectional regulators 19:42 bidirectional regulators of the melanocortin receptor family 19:78 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 0 3 theme family 73:78 arg1 MRAP 0:3 MRAP 0:3 MRAP 0:3 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 1 4 theme protein-coupled 136:150 arg1 MC1R-MC5R 163:171 MC1R-MC5R 163:171 MC1R-MC5R 163:171 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 1 4 theme protein-coupled 136:150 arg1 receptors 152:160 5 G protein-coupled receptors 132:160 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles 132:203 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 4 5 theme related 551:557 arg1 processes 541:549 disease processes 533:549 disease processes related to MCR dysfunction 533:576 Investigating the way in which these receptors signal and traffic to the cell membrane is vital in understanding disease processes related to MCR dysfunction. 19329486 5 6 theme adrenal 630:636 arg1 trafficking 643:653 adrenal MC2R trafficking 630:653 adrenal MC2R trafficking 630:653 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 5 7 theme MC2R 638:641 arg1 trafficking 643:653 adrenal MC2R trafficking 630:653 adrenal MC2R trafficking 630:653 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 9 8 theme [Nle4 976:980 arg1 hormone 1018:1024 [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone 976:1024 hormone 1018:1024 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 9 8 theme [Nle4 976:980 arg1 NDP-MSH 1027:1033 NDP-MSH 1027:1033 NDP-MSH 1027:1033 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 4 9 theme MCR 562:564 arg1 dysfunction 566:576 MCR dysfunction 562:576 MCR dysfunction 562:576 Investigating the way in which these receptors signal and traffic to the cell membrane is vital in understanding disease processes related to MCR dysfunction. 19329486 5 10 theme MC2R 590:593 arg1 protein 605:611 an MC2R accessory protein 587:611 an MC2R accessory protein 587:611 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 5 10 theme MC2R 590:593 arg1 MRAP 579:582 MRAP 579:582 MRAP 579:582 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 6 11 theme adrenal 750:756 arg1 gland 758:762 the adrenal gland 746:762 the adrenal gland 746:762 Here we identify MRAP2 as a unique homologue of MRAP, expressed in brain and the adrenal gland. 19329486 2 12 theme axis 273:276 arg1 component 225:233 a critical component 214:233 a critical component of the hypothalamic-pituitary-adrenal axis 214:276 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 2 12 theme axis 273:276 arg1 MC2R 206:209 MC2R 206:209 MC2R 206:209 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 2 13 dep component 225:233 arg1 whereas 279:285 whereas 279:285 whereas 279:285 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 10 14 theme bidirectional 1094:1106 arg1 regulators 1108:1117 unique bidirectional regulators 1087:1117 unique bidirectional regulators of the MCR family 1087:1135 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 14 theme bidirectional 1094:1106 arg1 MRAP2 1078:1082 MRAP2 1078:1082 MRAP2 1078:1082 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 14 theme bidirectional 1094:1106 arg1 MRAP 1069:1072 MRAP 1069:1072 MRAP 1069:1072 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 5 15 theme accessory 595:603 arg1 protein 605:611 an MC2R accessory protein 587:611 an MC2R accessory protein 587:611 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 5 15 theme accessory 595:603 arg1 MRAP 579:582 MRAP 579:582 MRAP 579:582 MRAP is an MC2R accessory protein, responsible for adrenal MC2R trafficking and function. 19329486 2 16 theme hypothalamic-pituitary-adrenal 242:271 arg1 axis 273:276 the hypothalamic-pituitary-adrenal axis 238:276 the hypothalamic-pituitary-adrenal axis 238:276 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 3 17 from Mutations 347:355 arg1 MC4R 360:363 MC4R 360:363 MC4R 360:363 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 1 18 theme melanocortin 85:96 arg1 MCR 108:110 MCR 108:110 MCR 108:110 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 1 18 theme melanocortin 85:96 arg1 receptor 98:105 melanocortin receptor 85:105 The melanocortin receptor (MCR) family 81:118 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 9 19 theme alpha-melanocyte-stimulating 989:1016 arg1 hormone 1018:1024 [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone 976:1024 hormone 1018:1024 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 9 19 theme alpha-melanocyte-stimulating 989:1016 arg1 NDP-MSH 1027:1033 NDP-MSH 1027:1033 NDP-MSH 1027:1033 In contrast, MRAP and MRAP2 can reduce MC1R, MC3R, MC4R, and MC5R responsiveness to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone (NDP-MSH). 19329486 3 20 theme monogenic 401:409 arg1 obesity 411:417 monogenic obesity 401:417 monogenic obesity 401:417 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 1 21 theme receptor 98:105 arg1 family 113:118 The melanocortin receptor (MCR) family 81:118 The melanocortin receptor (MCR) family 81:118 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 2 22 theme energy 327:332 arg1 homeostasis 334:344 energy homeostasis 327:344 energy homeostasis 327:344 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 1 23 theme diverse 179:185 arg1 roles 199:203 diverse physiologic roles 179:203 diverse physiologic roles 179:203 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 3 24 theme common 385:390 arg1 Mutations 347:355 Mutations 347:355 Mutations in MC4R 347:363 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 3 24 theme common 385:390 arg1 cause 392:396 the single most common cause 369:396 the single most common cause of monogenic obesity 369:417 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 4 25 theme disease 533:539 arg1 processes 541:549 disease processes 533:549 disease processes related to MCR dysfunction 533:576 Investigating the way in which these receptors signal and traffic to the cell membrane is vital in understanding disease processes related to MCR dysfunction. 19329486 1 26 theme physiologic 187:197 arg1 roles 199:203 diverse physiologic roles 179:203 diverse physiologic roles 179:203 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 0 27 theme bidirectional 19:31 arg1 MRAP2 9:13 MRAP2 9:13 MRAP2 9:13 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 0 27 theme bidirectional 19:31 arg1 regulators 33:42 bidirectional regulators 19:42 bidirectional regulators of the melanocortin receptor family 19:78 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 0 27 theme bidirectional 19:31 arg1 MRAP 0:3 MRAP 0:3 MRAP 0:3 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 8 28 theme surface 858:864 arg1 expression 866:875 MC2R surface expression 853:875 MC2R surface expression 853:875 This interaction results in MC2R surface expression and signaling. 19329486 3 29 theme single 373:378 arg1 Mutations 347:355 Mutations 347:355 Mutations in MC4R 347:363 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 3 29 theme single 373:378 arg1 cause 392:396 the single most common cause 369:396 the single most common cause of monogenic obesity 369:417 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 4 30 theme cell 493:496 arg1 membrane 498:505 the cell membrane 489:505 the cell membrane 489:505 Investigating the way in which these receptors signal and traffic to the cell membrane is vital in understanding disease processes related to MCR dysfunction. 19329486 3 31 theme obesity 411:417 arg1 Mutations 347:355 Mutations 347:355 Mutations in MC4R 347:363 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 3 31 theme obesity 411:417 arg1 cause 392:396 the single most common cause 369:396 the single most common cause of monogenic obesity 369:417 Mutations in MC4R are the single most common cause of monogenic obesity. 19329486 8 32 theme MC2R 853:856 arg1 expression 866:875 MC2R surface expression 853:875 MC2R surface expression 853:875 This interaction results in MC2R surface expression and signaling. 19329486 6 33 theme MRAP 717:720 arg1 homologue 704:712 a unique homologue 695:712 a unique homologue 695:712 Here we identify MRAP2 as a unique homologue of MRAP, expressed in brain and the adrenal gland. 19329486 6 33 theme MRAP 717:720 arg1 MRAP2 686:690 MRAP2 686:690 MRAP2 686:690 Here we identify MRAP2 as a unique homologue of MRAP, expressed in brain and the adrenal gland. 19329486 2 34 contain have 301:304 arg1 MC4R 296:299 MC4R 296:299 MC4R 296:299 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 2 34 contain have 301:304 arg1 MC3R 287:290 MC3R 287:290 MC3R 287:290 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 2 34 contain have 301:304 arg2 role 319:322 an essential role 306:322 an essential role 306:322 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 1 35 with receptors 152:160 arg1 roles 199:203 diverse physiologic roles 179:203 diverse physiologic roles 179:203 The melanocortin receptor (MCR) family consists of 5 G protein-coupled receptors (MC1R-MC5R) with diverse physiologic roles. 19329486 2 36 theme essential 309:317 arg1 role 319:322 an essential role 306:322 an essential role 306:322 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 10 37 theme unique 1087:1092 arg1 regulators 1108:1117 unique bidirectional regulators 1087:1117 unique bidirectional regulators of the MCR family 1087:1135 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 37 theme unique 1087:1092 arg1 MRAP2 1078:1082 MRAP2 1078:1082 MRAP2 1078:1082 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 37 theme unique 1087:1092 arg1 MRAP 1069:1072 MRAP 1069:1072 MRAP 1069:1072 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 38 theme family 1130:1135 arg1 regulators 1108:1117 unique bidirectional regulators 1087:1117 unique bidirectional regulators of the MCR family 1087:1135 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 38 theme family 1130:1135 arg1 MRAP2 1078:1082 MRAP2 1078:1082 MRAP2 1078:1082 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 10 38 theme family 1130:1135 arg1 MRAP 1069:1072 MRAP 1069:1072 MRAP 1069:1072 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 0 39 theme receptor 64:71 arg1 family 73:78 the melanocortin receptor family 47:78 the melanocortin receptor family 47:78 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 6 40 theme unique 697:702 arg1 homologue 704:712 a unique homologue 695:712 a unique homologue 695:712 Here we identify MRAP2 as a unique homologue of MRAP, expressed in brain and the adrenal gland. 19329486 6 40 theme unique 697:702 arg1 MRAP2 686:690 MRAP2 686:690 MRAP2 686:690 Here we identify MRAP2 as a unique homologue of MRAP, expressed in brain and the adrenal gland. 19329486 10 41 theme MCR 1126:1128 arg1 family 1130:1135 the MCR family 1122:1135 the MCR family 1122:1135 Collectively, our data identify MRAP and MRAP2 as unique bidirectional regulators of the MCR family. 19329486 0 42 theme melanocortin 51:62 arg1 family 73:78 the melanocortin receptor family 47:78 the melanocortin receptor family 47:78 MRAP and MRAP2 are bidirectional regulators of the melanocortin receptor family. 19329486 2 43 theme critical 216:223 arg1 component 225:233 a critical component 214:233 a critical component of the hypothalamic-pituitary-adrenal axis 214:276 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 19329486 2 43 theme critical 216:223 arg1 MC2R 206:209 MC2R 206:209 MC2R 206:209 MC2R is a critical component of the hypothalamic-pituitary-adrenal axis, whereas MC3R and MC4R have an essential role in energy homeostasis. 27399812 5 0 theme efficient 735:743 arg1 analysis 745:752 efficient analysis 735:752 efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra 735:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 6 1 theme chondroitin 931:941 arg1 sulfate 943:949 chondroitin sulfate 931:949 chondroitin sulfate 931:949 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 9 2 theme chondroitin 1578:1588 arg1 proteoglycans 1598:1610 chondroitin sulfate proteoglycans 1578:1610 chondroitin sulfate proteoglycans 1578:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 3 theme classes 1311:1317 arg1 assignment 1260:1269 the initial assignment 1248:1269 the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms 1248:1349 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 6 4 theme glycopeptides 913:925 arg1 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 2 5 theme time-consuming 318:331 arg1 interpretations 340:354 time-consuming manual interpretations 318:354 time-consuming manual interpretations 318:354 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 9 6 theme proteoglycans 1598:1610 arg1 region 1568:1573 the linkage region 1556:1573 the linkage region of chondroitin sulfate proteoglycans 1556:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 7 theme clustering 1130:1139 arg1 combination 1106:1116 The combination 1102:1116 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions 1102:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 5 8 theme data-oriented 693:705 arg1 workflow 722:729 a data-oriented bioinformatics workflow 691:729 SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra 681:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 1 9 theme proteomics 185:194 arg1 fields 161:166 the fields 157:166 the fields of glycomics and proteomics 157:194 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 8 10 theme glycoforms 1340:1349 arg1 assignment 1260:1269 the initial assignment 1248:1269 the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms 1248:1349 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 8 11 theme oxonium 1142:1148 arg1 profiles 1164:1171 oxonium ion intensity profiles 1142:1171 oxonium ion intensity profiles 1142:1171 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 1 12 gly glycosylation 229:241 arg1 biology 261:267 biology 261:267 biology 261:267 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 6 13 theme enrichment 871:880 arg1 protocols 882:890 two separate glycopeptide enrichment protocols 845:890 two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 845:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 8 14 theme intensity 1154:1162 arg1 profiles 1164:1171 oxonium ion intensity profiles 1142:1171 oxonium ion intensity profiles 1142:1171 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 5 15 theme MS/MS-spectra 795:807 arg1 thousands 769:777 thousands 769:777 thousands of glycopeptide MS/MS-spectra 769:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 1 16 theme protein 221:227 arg1 glycosylation 229:241 site-specific protein glycosylation 207:241 site-specific protein glycosylation 207:241 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 9 17 theme database 1396:1403 arg1 searches 1405:1412 database searches 1396:1412 database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1396:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 1 18 from glycosylation 229:241 arg1 biology 261:267 biology 261:267 biology 261:267 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 1 19 theme analytical 128:137 arg1 platform 139:146 an independent analytical platform 113:146 an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology 113:267 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 6 20 gly glycopeptides 913:925 arg2 glycopeptides 913:925 glycopeptides 913:925 glycopeptides 913:925 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 7 21 gly glycopeptide 1046:1057 arg2 glycopeptide 1046:1057 the glycopeptide MS/MS data 1042:1068 the glycopeptide MS/MS data based on spectral similarities 1042:1099 Molecular networking was performed to organize the glycopeptide MS/MS data based on spectral similarities. 27399812 1 22 from impact 251:256 arg1 biology 261:267 biology 261:267 biology 261:267 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 2 23 theme glycopeptide 278:289 arg1 characterization 291:306 Current glycopeptide characterization 270:306 Current glycopeptide characterization 270:306 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 0 24 theme Spectral 59:66 arg1 Analysis 68:75 Glycopeptide MS/MS Spectral Analysis 40:75 Glycopeptide MS/MS Spectral Analysis 40:75 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 27399812 9 25 gly glycopeptide 1480:1491 arg2 glycopeptide 1480:1491 glycopeptide 1480:1491 glycopeptide 1480:1491 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 9 26 from modifications 1539:1551 arg1 region 1568:1573 the linkage region 1556:1573 the linkage region of chondroitin sulfate proteoglycans 1556:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 1 27 theme glycomics 171:179 arg1 fields 161:166 the fields 157:166 the fields of glycomics and proteomics 157:194 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 3 28 theme data 413:416 arg1 interpretation 418:431 Efficient data interpretation 403:431 Efficient data interpretation 403:431 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 4 29 theme glyco-related 578:590 arg1 tools 607:611 new glyco-related bioinformatics tools 574:611 new glyco-related bioinformatics tools 574:611 The development of new glyco-related bioinformatics tools is thus of crucial importance to fulfill this goal. 27399812 9 30 used used 1382:1385 arg2 signatures 1358:1367 These signatures 1352:1367 These signatures 1352:1367 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 9 31 theme large 1464:1468 arg1 number 1470:1475 a large number 1462:1475 a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1462:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 9 31 theme large 1464:1468 arg1 modifications 1539:1551 novel deoxyhexose (fucose) modifications 1512:1551 novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1512:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 32 theme precursor 1178:1186 arg1 distributions 1202:1214 precursor ion m/z shift distributions 1178:1214 precursor ion m/z shift distributions 1178:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 6 33 gly glycopeptides 966:978 arg2 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 3 34 theme true 495:498 arg1 analysis 529:536 true high-throughput glycopeptide analysis 495:536 true high-throughput glycopeptide analysis 495:536 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 8 35 theme m/z 1192:1194 arg1 distributions 1202:1214 precursor ion m/z shift distributions 1178:1214 precursor ion m/z shift distributions 1178:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 9 36 theme variants 1493:1500 arg1 number 1470:1475 a large number 1462:1475 a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1462:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 9 36 theme variants 1493:1500 arg1 modifications 1539:1551 novel deoxyhexose (fucose) modifications 1512:1551 novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1512:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 37 theme distributions 1202:1214 arg1 combination 1106:1116 The combination 1102:1116 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions 1102:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 4 38 theme crucial 624:630 arg1 importance 632:641 crucial importance 624:641 crucial importance 624:641 The development of new glyco-related bioinformatics tools is thus of crucial importance to fulfill this goal. 27399812 9 39 theme novel 1512:1516 arg1 modifications 1539:1551 novel deoxyhexose (fucose) modifications 1512:1551 novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1512:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 40 theme typical 1225:1231 arg1 signatures 1233:1242 typical signatures 1225:1242 typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms 1225:1349 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 3 41 theme glycopeptide 516:527 arg1 analysis 529:536 true high-throughput glycopeptide analysis 495:536 true high-throughput glycopeptide analysis 495:536 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 6 42 from protocols 882:890 arg1 sets 835:838 MS data sets 827:838 MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 827:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 3 43 theme major 456:460 arg1 challenges 462:471 the major challenges 452:471 the major challenges 452:471 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 4 44 theme tools 607:611 arg1 development 559:569 The development 555:569 The development of new glyco-related bioinformatics tools 555:611 The development of new glyco-related bioinformatics tools is thus of crucial importance to fulfill this goal. 27399812 2 45 theme expertise 392:400 arg1 levels 373:378 high levels 368:378 high levels of personal expertise 368:400 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 0 46 theme Glycopeptide 40:51 arg1 Analysis 68:75 Glycopeptide MS/MS Spectral Analysis 40:75 Glycopeptide MS/MS Spectral Analysis 40:75 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 27399812 8 47 theme initial 1252:1258 arg1 assignment 1260:1269 the initial assignment 1248:1269 the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms 1248:1349 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 5 48 dep SweetNET 681:688 arg1 workflow 722:729 a data-oriented bioinformatics workflow 691:729 SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra 681:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 7 49 theme MS/MS 1059:1063 arg1 data 1065:1068 the glycopeptide MS/MS data 1042:1068 the glycopeptide MS/MS data based on spectral similarities 1042:1099 Molecular networking was performed to organize the glycopeptide MS/MS data based on spectral similarities. 27399812 6 50 theme linkage 951:957 arg1 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 6 51 theme MS 827:828 arg1 sets 835:838 MS data sets 827:838 MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 827:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 9 52 theme linkage 1560:1566 arg1 region 1568:1573 the linkage region 1556:1573 the linkage region of chondroitin sulfate proteoglycans 1556:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 53 theme N- 1284:1285 arg1 classes 1311:1317 different N-, O- and CS-glycopeptide classes 1274:1317 different N-, O- and CS-glycopeptide classes 1274:1317 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 0 54 dep SweetNET 0:7 arg1 Workflow 27:34 A Bioinformatics Workflow 10:34 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 0:76 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 27399812 8 55 theme O- 1288:1289 arg1 classes 1311:1317 different N-, O- and CS-glycopeptide classes 1274:1317 different N-, O- and CS-glycopeptide classes 1274:1317 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 8 56 theme spectral 1121:1128 arg1 clustering 1130:1139 spectral clustering 1121:1139 spectral clustering 1121:1139 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 9 57 dep identification 1429:1442 arg1 the 1425:1427 the 1425:1427 the 1425:1427 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 2 58 theme manual 333:338 arg1 interpretations 340:354 time-consuming manual interpretations 318:354 time-consuming manual interpretations 318:354 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 9 59 theme sulfate 1590:1596 arg1 proteoglycans 1598:1610 chondroitin sulfate proteoglycans 1578:1610 chondroitin sulfate proteoglycans 1578:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 60 theme CS-glycopeptide 1295:1309 arg1 classes 1311:1317 different N-, O- and CS-glycopeptide classes 1274:1317 different N-, O- and CS-glycopeptide classes 1274:1317 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 6 61 theme sialylated 902:911 arg1 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 7 62 theme Molecular 995:1003 arg1 networking 1005:1014 Molecular networking 995:1014 Molecular networking 995:1014 Molecular networking was performed to organize the glycopeptide MS/MS data based on spectral similarities. 27399812 8 63 theme ion 1150:1152 arg1 profiles 1164:1171 oxonium ion intensity profiles 1142:1171 oxonium ion intensity profiles 1142:1171 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 2 64 gly glycopeptide 278:289 arg2 glycopeptide 278:289 Current glycopeptide characterization 270:306 Current glycopeptide characterization 270:306 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 5 65 theme bioinformatics 707:720 arg1 workflow 722:729 a data-oriented bioinformatics workflow 691:729 SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra 681:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 2 66 theme high 368:371 arg1 levels 373:378 high levels 368:378 high levels of personal expertise 368:400 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 8 67 theme respective 1329:1338 arg1 glycoforms 1340:1349 their respective glycoforms 1323:1349 their respective glycoforms 1323:1349 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 8 68 theme profiles 1164:1171 arg1 combination 1106:1116 The combination 1102:1116 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions 1102:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 5 69 theme glycopeptide 782:793 arg1 MS/MS-spectra 795:807 glycopeptide MS/MS-spectra 782:807 glycopeptide MS/MS-spectra 782:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 6 70 theme glycopeptide 858:869 arg1 protocols 882:890 two separate glycopeptide enrichment protocols 845:890 two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 845:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 1 71 theme site-specific 207:219 arg1 glycosylation 229:241 site-specific protein glycosylation 207:241 site-specific protein glycosylation 207:241 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 6 72 gly glycopeptide 858:869 arg2 glycopeptide 858:869 two separate glycopeptide enrichment protocols 845:890 two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 845:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 5 73 theme hundreds 757:764 arg1 analysis 745:752 efficient analysis 735:752 efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra 735:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 1 74 theme independent 116:126 arg1 platform 139:146 an independent analytical platform 113:146 an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology 113:267 Glycoproteomics has rapidly become an independent analytical platform bridging the fields of glycomics and proteomics to address site-specific protein glycosylation and its impact in biology. 27399812 6 75 gly sialylated 902:911 arg1 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 5 76 gly glycopeptide 782:793 arg2 glycopeptide 782:793 glycopeptide MS/MS-spectra 782:807 glycopeptide MS/MS-spectra 782:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 5 77 theme thousands 769:777 arg1 hundreds 757:764 hundreds 757:764 hundreds of thousands of glycopeptide MS/MS-spectra 757:807 Here we present SweetNET: a data-oriented bioinformatics workflow for efficient analysis of hundreds of thousands of glycopeptide MS/MS-spectra. 27399812 2 78 theme Current 270:276 arg1 characterization 291:306 Current glycopeptide characterization 270:306 Current glycopeptide characterization 270:306 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 4 79 theme new 574:576 arg1 tools 607:611 new glyco-related bioinformatics tools 574:611 new glyco-related bioinformatics tools 574:611 The development of new glyco-related bioinformatics tools is thus of crucial importance to fulfill this goal. 27399812 4 80 theme bioinformatics 592:605 arg1 tools 607:611 new glyco-related bioinformatics tools 574:611 new glyco-related bioinformatics tools 574:611 The development of new glyco-related bioinformatics tools is thus of crucial importance to fulfill this goal. 27399812 8 81 theme ion 1188:1190 arg1 distributions 1202:1214 precursor ion m/z shift distributions 1178:1214 precursor ion m/z shift distributions 1178:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 6 82 theme data 830:833 arg1 sets 835:838 MS data sets 827:838 MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 827:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 9 83 theme number 1470:1475 arg1 validation 1448:1457 validation 1448:1457 validation 1448:1457 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 9 83 theme number 1470:1475 arg1 identification 1429:1442 identification 1429:1442 identification 1429:1442 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 8 84 theme shift 1196:1200 arg1 distributions 1202:1214 precursor ion m/z shift distributions 1178:1214 precursor ion m/z shift distributions 1178:1214 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 3 85 theme Efficient 403:411 arg1 interpretation 418:431 Efficient data interpretation 403:431 Efficient data interpretation 403:431 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 3 86 theme high-throughput 500:514 arg1 analysis 529:536 true high-throughput glycopeptide analysis 495:536 true high-throughput glycopeptide analysis 495:536 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 9 87 theme glycopeptide 1480:1491 arg1 variants 1493:1500 glycopeptide variants 1480:1500 glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1480:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 9 87 theme glycopeptide 1480:1491 arg1 modifications 1539:1551 novel deoxyhexose (fucose) modifications 1512:1551 novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1512:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 7 88 theme spectral 1079:1086 arg1 similarities 1088:1099 spectral similarities 1079:1099 spectral similarities 1079:1099 Molecular networking was performed to organize the glycopeptide MS/MS data based on spectral similarities. 27399812 0 89 theme Bioinformatics 12:25 arg1 Workflow 27:34 A Bioinformatics Workflow 10:34 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 0:76 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 27399812 9 90 theme deoxyhexose 1518:1528 arg1 modifications 1539:1551 novel deoxyhexose (fucose) modifications 1512:1551 novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1512:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 6 91 theme separate 849:856 arg1 protocols 882:890 two separate glycopeptide enrichment protocols 845:890 two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively 845:992 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 0 92 theme MS/MS 53:57 arg1 Analysis 68:75 Glycopeptide MS/MS Spectral Analysis 40:75 Glycopeptide MS/MS Spectral Analysis 40:75 SweetNET: A Bioinformatics Workflow for Glycopeptide MS/MS Spectral Analysis. 27399812 9 93 theme fucose 1531:1536 arg1 modifications 1539:1551 novel deoxyhexose (fucose) modifications 1512:1551 novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans 1512:1610 These signatures were further used to guide database searches leading to the identification and validation of a large number of glycopeptide variants including novel deoxyhexose (fucose) modifications in the linkage region of chondroitin sulfate proteoglycans. 27399812 3 94 theme challenges 462:471 arg1 challenges 462:471 the major challenges 452:471 the major challenges 452:471 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 3 94 theme challenges 462:471 arg1 one 445:447 one 445:447 one 445:447 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 3 95 gly glycopeptide 516:527 arg2 glycopeptide 516:527 true high-throughput glycopeptide analysis 495:536 true high-throughput glycopeptide analysis 495:536 Efficient data interpretation constitutes one of the major challenges to be overcome before true high-throughput glycopeptide analysis can be achieved. 27399812 2 96 theme personal 383:390 arg1 expertise 392:400 personal expertise 383:400 personal expertise 383:400 Current glycopeptide characterization relies on time-consuming manual interpretations and demands high levels of personal expertise. 27399812 7 97 theme glycopeptide 1046:1057 arg1 data 1065:1068 the glycopeptide MS/MS data 1042:1068 the glycopeptide MS/MS data based on spectral similarities 1042:1099 Molecular networking was performed to organize the glycopeptide MS/MS data based on spectral similarities. 27399812 8 98 theme different 1274:1282 arg1 N- 1284:1285 different N- 1274:1285 different N- 1274:1285 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 6 99 theme region 959:964 arg1 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 27399812 8 100 gly CS-glycopeptide 1295:1309 arg2 CS-glycopeptide 1295:1309 CS-glycopeptide 1295:1309 CS-glycopeptide 1295:1309 The combination of spectral clustering, oxonium ion intensity profiles, and precursor ion m/z shift distributions provided typical signatures for the initial assignment of different N-, O- and CS-glycopeptide classes and their respective glycoforms. 27399812 6 101 theme sulfate 943:949 arg1 glycopeptides 966:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides 902:978 We have analyzed MS data sets from two separate glycopeptide enrichment protocols targeting sialylated glycopeptides and chondroitin sulfate linkage region glycopeptides, respectively. 15651049 9 0 theme competent 1613:1621 arg1 juxtaposition 1623:1635 a catalytically competent juxtaposition 1597:1635 a catalytically competent juxtaposition with the active-site His 57 1597:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 6 1 theme recombinant 1072:1082 arg1 K1 1094:1095 mature, active recombinant human apo K1 1057:1095 mature, active recombinant human apo K1 1057:1095 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 2 2 theme blood 374:378 arg1 pressure 380:387 blood pressure 374:387 blood pressure 374:387 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 3 3 theme prostate 556:563 arg1 cancer 565:570 prostate cancer 556:570 prostate cancer 556:570 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 9 4 theme active-site 1646:1656 arg1 His 1658:1660 the active-site His 57 1642:1663 the active-site His 57 1642:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 2 5 theme degradative-type 275:290 arg1 proteases 292:300 both regulatory- and degradative-type proteases 254:300 both regulatory- and degradative-type proteases 254:300 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 4 6 theme pressure 827:834 arg1 regulation 836:845 blood pressure regulation 821:845 blood pressure regulation 821:845 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 1 7 theme large 186:190 arg1 family 222:227 a recently identified large and closely related 15-member family 164:227 a recently identified large and closely related 15-member family 164:227 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 9 8 with juxtaposition 1623:1635 arg1 His 1658:1660 the active-site His 57 1642:1663 the active-site His 57 1642:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 1 9 theme 15-member 212:220 arg1 family 222:227 a recently identified large and closely related 15-member family 164:227 a recently identified large and closely related 15-member family 164:227 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 6 10 theme apo 1090:1092 arg1 K1 1094:1095 mature, active recombinant human apo K1 1057:1095 mature, active recombinant human apo K1 1057:1095 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 10 11 dep Arg-Ser 1733:1739 arg1 specificity 1768:1778 dual substrate specificity 1753:1778 dual substrate specificity 1753:1778 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 10 11 dep Arg-Ser 1733:1739 arg1 the 1729:1731 the 1729:1731 the 1729:1731 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 10 12 theme dual 1753:1756 arg1 specificity 1768:1778 dual substrate specificity 1753:1778 dual substrate specificity 1753:1778 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 1 13 theme Human 111:115 arg1 kallikreins 117:127 Human kallikreins 111:127 Human kallikreins 111:127 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 1 13 theme Human 111:115 arg1 proteases 140:148 serine proteases 133:148 serine proteases that comprise a recently identified large and closely related 15-member family 133:227 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 8 14 theme extensive 1425:1433 arg1 network 1443:1449 an extensive solvent network 1422:1449 an extensive solvent network 1422:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 2 15 theme regulatory- 259:269 arg1 proteases 292:300 both regulatory- and degradative-type proteases 254:300 both regulatory- and degradative-type proteases 254:300 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 5 16 theme unique 892:897 arg1 specificity 914:924 unique dual-substrate specificity 892:924 unique dual-substrate specificity 892:924 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 8 17 theme network 1443:1449 arg1 displacement 1406:1417 the displacement 1402:1417 the displacement of an extensive solvent network 1402:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 4 18 theme tissue 688:693 arg1 kallikrein 695:704 Human tissue kallikrein 682:704 Human tissue kallikrein (human K1) 682:715 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 18 theme tissue 688:693 arg1 K1 713:714 human K1 707:714 human K1 707:714 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 18 theme tissue 688:693 arg1 member 756:761 the best functionally characterized member 720:761 the best functionally characterized member of this family 720:776 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 3 19 theme breast 629:634 arg1 cancer 636:641 breast cancer 629:641 breast cancer 629:641 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 8 20 theme conformational 1290:1303 arg1 changes 1305:1311 conformational changes 1290:1311 conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1290:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 0 21 theme structural 50:59 arg1 changes 61:67 structural changes 50:67 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 0:109 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 5 22 theme molecular 945:953 arg1 kininogen 962:970 low molecular weight kininogen 941:970 low molecular weight kininogen between both Arg-Ser and Met-Lys sequences 941:1013 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 4 23 theme family 771:776 arg1 kallikrein 695:704 Human tissue kallikrein 682:704 Human tissue kallikrein (human K1) 682:715 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 23 theme family 771:776 arg1 member 756:761 the best functionally characterized member 720:761 the best functionally characterized member of this family 720:776 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 0 24 theme inhibitor/substrate 82:100 arg1 binding 102:108 peptide inhibitor/substrate binding 74:108 peptide inhibitor/substrate binding 74:108 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 6 25 theme mature 1057:1062 arg1 K1 1094:1095 mature, active recombinant human apo K1 1057:1095 mature, active recombinant human apo K1 1057:1095 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 8 26 theme porcine 1334:1340 arg1 homolog 1342:1348 the porcine homolog 1330:1348 the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1330:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 0 27 theme 1.70 0:3 arg1 structure 13:21 1.70 A X-ray structure 0:21 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 0:109 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 8 28 from variety 1279:1285 arg1 comparison 1316:1325 comparison 1316:1325 comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1316:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 9 29 theme structural 1532:1541 arg1 rearrangement 1543:1555 a structural rearrangement 1530:1555 a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57 1530:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 4 30 theme characterized 742:754 arg1 kallikrein 695:704 Human tissue kallikrein 682:704 Human tissue kallikrein (human K1) 682:715 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 30 theme characterized 742:754 arg1 member 756:761 the best functionally characterized member 720:761 the best functionally characterized member of this family 720:776 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 6 31 from resolution 1107:1116 arg1 structure 1044:1052 the X-ray crystal structure 1026:1052 the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution 1026:1116 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 0 32 theme X-ray 7:11 arg1 structure 13:21 1.70 A X-ray structure 0:21 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 0:109 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 6 33 theme X-ray 1030:1034 arg1 structure 1044:1052 the X-ray crystal structure 1026:1052 the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution 1026:1116 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 3 34 theme majority 464:471 arg1 function 448:455 the function 444:455 the function of the majority of the kallikreins 444:490 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 7 35 theme active 1123:1128 arg1 site 1130:1133 The active site 1119:1133 The active site 1119:1133 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 9 36 theme active-site 1564:1574 arg1 195 1580:1582 the active-site Ser 195 1560:1582 the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57 1560:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 4 37 theme important 803:811 arg1 role 813:816 an important role 800:816 an important role 800:816 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 0 38 theme apo 32:34 arg1 kallikrein 36:45 human apo kallikrein 1 26:47 human apo kallikrein 1 26:47 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 9 39 theme 195 1580:1582 arg1 rearrangement 1543:1555 a structural rearrangement 1530:1555 a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57 1530:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 10 40 theme human 1783:1787 arg1 K1 1789:1790 human K1 1783:1790 human K1 1783:1790 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 10 41 theme solvent 1670:1676 arg1 networks 1678:1685 The solvent networks 1666:1685 The solvent networks within the S1 and S1' pockets 1666:1715 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 8 42 from comparison 1316:1325 arg1 variety 1279:1285 a variety 1277:1285 a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1277:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 8 42 from comparison 1316:1325 arg1 changes 1305:1311 conformational changes 1290:1311 conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1290:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 6 43 theme K1 1094:1095 arg1 structure 1044:1052 the X-ray crystal structure 1026:1052 the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution 1026:1116 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 2 44 theme neuronal 390:397 arg1 health 399:404 neuronal health 390:404 neuronal health 390:404 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 8 45 from changes 1305:1311 arg1 comparison 1316:1325 comparison 1316:1325 comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1316:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 6 46 theme human 1084:1088 arg1 K1 1094:1095 mature, active recombinant human apo K1 1057:1095 mature, active recombinant human apo K1 1057:1095 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 2 47 theme pressure 380:387 arg1 regulation 360:369 regulation 360:369 regulation of blood pressure 360:387 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 47 theme pressure 380:387 arg1 response 428:435 the inflammatory response 411:435 the inflammatory response 411:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 47 theme pressure 380:387 arg1 health 399:404 neuronal health 390:404 neuronal health 390:404 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 8 48 from homolog 1342:1348 arg1 complex 1359:1365 complex 1359:1365 complex with peptide inhibitors, including the displacement of an extensive solvent network 1359:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 7 49 theme known 1214:1218 arg1 structures 1231:1240 known kallikrein structures 1214:1240 known kallikrein structures 1214:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 2 50 theme physiological 326:338 arg1 regulation 360:369 regulation 360:369 regulation of blood pressure 360:387 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 50 theme physiological 326:338 arg1 response 428:435 the inflammatory response 411:435 the inflammatory response 411:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 50 theme physiological 326:338 arg1 processes 340:348 physiological processes 326:348 physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response 326:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 50 theme physiological 326:338 arg1 health 399:404 neuronal health 390:404 neuronal health 390:404 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 3 51 theme useful 534:539 arg1 members 522:528 two members 518:528 two members 518:528 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 3 51 theme useful 534:539 arg1 biomarkers 541:550 useful biomarkers 534:550 useful biomarkers for prostate cancer 534:570 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 8 52 from complex 1359:1365 arg1 homolog 1342:1348 the porcine homolog 1330:1348 the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1330:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 3 53 theme several 576:582 arg1 biomarkers 614:623 potentially useful biomarkers 595:623 potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease 595:679 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 3 53 theme several 576:582 arg1 others 584:589 several others 576:589 several others 576:589 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 1 54 theme identified 175:184 arg1 family 222:227 a recently identified large and closely related 15-member family 164:227 a recently identified large and closely related 15-member family 164:227 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 4 55 theme blood 821:825 arg1 regulation 836:845 blood pressure regulation 821:845 blood pressure regulation 821:845 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 8 56 with complex 1359:1365 arg1 inhibitors 1380:1389 peptide inhibitors 1372:1389 peptide inhibitors 1372:1389 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 8 56 with complex 1359:1365 arg1 displacement 1406:1417 the displacement 1402:1417 the displacement of an extensive solvent network 1402:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 3 57 theme useful 607:612 arg1 biomarkers 614:623 potentially useful biomarkers 595:623 potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease 595:679 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 3 57 theme useful 607:612 arg1 others 584:589 several others 576:589 several others 576:589 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 1 58 theme related 204:210 arg1 family 222:227 a recently identified large and closely related 15-member family 164:227 a recently identified large and closely related 15-member family 164:227 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 10 59 theme substrate 1758:1766 arg1 specificity 1768:1778 dual substrate specificity 1753:1778 dual substrate specificity 1753:1778 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 5 60 theme human 874:878 arg1 K1 880:881 human K1 874:881 human K1 874:881 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 8 61 theme solvent 1435:1441 arg1 network 1443:1449 an extensive solvent network 1422:1449 an extensive solvent network 1422:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 4 62 theme human 707:711 arg1 kallikrein 695:704 Human tissue kallikrein 682:704 Human tissue kallikrein (human K1) 682:715 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 62 theme human 707:711 arg1 K1 713:714 human K1 707:714 human K1 707:714 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 9 63 theme peptide 1497:1503 arg1 substrate 1505:1513 a peptide substrate 1495:1513 a peptide substrate 1495:1513 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 7 64 theme pro 1201:1203 arg1 forms 1205:1209 apo and pro forms 1193:1209 apo and pro forms of known kallikrein structures 1193:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 10 65 theme K1 1789:1790 arg1 Arg-Ser 1733:1739 Arg-Ser 1733:1739 Arg-Ser 1733:1739 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 10 65 theme K1 1789:1790 arg1 Met-Lys 1745:1751 Met-Lys 1745:1751 Met-Lys 1745:1751 The solvent networks within the S1 and S1' pockets suggest how the Arg-Ser and Met-Lys dual substrate specificity of human K1 is accommodated. 15651049 1 66 theme serine 133:138 arg1 kallikreins 117:127 Human kallikreins 111:127 Human kallikreins 111:127 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 1 66 theme serine 133:138 arg1 proteases 140:148 serine proteases 133:148 serine proteases that comprise a recently identified large and closely related 15-member family 133:227 Human kallikreins are serine proteases that comprise a recently identified large and closely related 15-member family. 15651049 4 67 theme Human 682:686 arg1 kallikrein 695:704 Human tissue kallikrein 682:704 Human tissue kallikrein (human K1) 682:715 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 67 theme Human 682:686 arg1 K1 713:714 human K1 707:714 human K1 707:714 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 4 67 theme Human 682:686 arg1 member 756:761 the best functionally characterized member 720:761 the best functionally characterized member of this family 720:776 Human tissue kallikrein (human K1) is the best functionally characterized member of this family, and is known to play an important role in blood pressure regulation. 15651049 7 68 theme apo 1193:1195 arg1 forms 1205:1209 apo and pro forms 1193:1209 apo and pro forms of known kallikrein structures 1193:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 5 69 theme function 864:871 arg1 part 851:854 part 851:854 part of this function 851:871 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 0 70 theme peptide 74:80 arg1 binding 102:108 peptide inhibitor/substrate binding 74:108 peptide inhibitor/substrate binding 74:108 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 5 71 theme low 941:943 arg1 kininogen 962:970 low molecular weight kininogen 941:970 low molecular weight kininogen between both Arg-Ser and Met-Lys sequences 941:1013 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 9 72 theme substrate 1505:1513 arg1 binding 1484:1490 the binding 1480:1490 the binding of a peptide substrate 1480:1513 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 8 73 theme changes 1305:1311 arg1 variety 1279:1285 a variety 1277:1285 a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1277:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 8 73 theme changes 1305:1311 arg1 changes 1305:1311 conformational changes 1290:1311 conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1290:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 2 74 theme processes 340:348 arg1 regulation 360:369 regulation 360:369 regulation of blood pressure 360:387 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 74 theme processes 340:348 arg1 variety 315:321 a variety 313:321 a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response 313:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 74 theme processes 340:348 arg1 processes 340:348 physiological processes 326:348 physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response 326:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 74 theme processes 340:348 arg1 health 399:404 neuronal health 390:404 neuronal health 390:404 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 2 74 theme processes 340:348 arg1 response 428:435 the inflammatory response 411:435 the inflammatory response 411:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 7 75 theme intermediate 1164:1175 arg1 features 1155:1162 structural features 1144:1162 structural features intermediate between that of apo and pro forms of known kallikrein structures 1144:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 5 76 theme weight 955:960 arg1 kininogen 962:970 low molecular weight kininogen 941:970 low molecular weight kininogen between both Arg-Ser and Met-Lys sequences 941:1013 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 0 77 theme A 5:5 arg1 structure 13:21 1.70 A X-ray structure 0:21 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 0:109 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 5 78 theme dual-substrate 899:912 arg1 specificity 914:924 unique dual-substrate specificity 892:924 unique dual-substrate specificity 892:924 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 6 79 theme crystal 1036:1042 arg1 structure 1044:1052 the X-ray crystal structure 1026:1052 the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution 1026:1116 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 6 80 dep mature 1057:1062 arg1 active 1065:1070 active 1065:1070 active 1065:1070 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 3 81 theme kallikreins 480:490 arg1 majority 464:471 the majority 460:471 the majority of the kallikreins 460:490 While the function of the majority of the kallikreins remains to be elucidated, two members are useful biomarkers for prostate cancer and several others are potentially useful biomarkers for breast cancer, Alzheimer's, and Parkinson's disease. 15651049 0 82 theme human 26:30 arg1 kallikrein 36:45 human apo kallikrein 1 26:47 human apo kallikrein 1 26:47 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 5 83 theme Met-Lys 997:1003 arg1 sequences 1005:1013 both Arg-Ser and Met-Lys sequences 980:1013 both Arg-Ser and Met-Lys sequences 980:1013 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 8 84 theme K1 1353:1354 arg1 homolog 1342:1348 the porcine homolog 1330:1348 the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network 1330:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 2 85 theme inflammatory 415:426 arg1 response 428:435 the inflammatory response 411:435 the inflammatory response 411:435 The kallikreins include both regulatory- and degradative-type proteases, impacting a variety of physiological processes including regulation of blood pressure, neuronal health, and the inflammatory response. 15651049 0 86 theme kallikrein 36:45 arg1 structure 13:21 1.70 A X-ray structure 0:21 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 0:109 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 9 87 theme Ser 1576:1578 arg1 195 1580:1582 the active-site Ser 195 1560:1582 the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57 1560:1663 These results indicate that the binding of a peptide substrate contributes to a structural rearrangement of the active-site Ser 195 resulting in a catalytically competent juxtaposition with the active-site His 57. 15651049 6 88 theme A 1105:1105 arg1 resolution 1107:1116 1.70 A resolution 1100:1116 1.70 A resolution 1100:1116 We report the X-ray crystal structure of mature, active recombinant human apo K1 at 1.70 A resolution. 15651049 0 89 dep structure 13:21 arg1 changes 61:67 structural changes 50:67 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 0:109 1.70 A X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. 15651049 8 90 theme peptide 1372:1378 arg1 inhibitors 1380:1389 peptide inhibitors 1372:1389 peptide inhibitors 1372:1389 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 8 90 theme peptide 1372:1378 arg1 displacement 1406:1417 the displacement 1402:1417 the displacement of an extensive solvent network 1402:1449 The S2 to S2' pockets demonstrate a variety of conformational changes in comparison to the porcine homolog of K1 in complex with peptide inhibitors, including the displacement of an extensive solvent network. 15651049 7 91 theme structures 1231:1240 arg1 forms 1205:1209 apo and pro forms 1193:1209 apo and pro forms of known kallikrein structures 1193:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 5 92 theme Arg-Ser 985:991 arg1 sequences 1005:1013 both Arg-Ser and Met-Lys sequences 980:1013 both Arg-Ser and Met-Lys sequences 980:1013 As part of this function, human K1 exhibits unique dual-substrate specificity in hydrolyzing low molecular weight kininogen between both Arg-Ser and Met-Lys sequences. 15651049 7 93 theme kallikrein 1220:1229 arg1 structures 1231:1240 known kallikrein structures 1214:1240 known kallikrein structures 1214:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 15651049 7 94 theme structural 1144:1153 arg1 features 1155:1162 structural features 1144:1162 structural features intermediate between that of apo and pro forms of known kallikrein structures 1144:1240 The active site exhibits structural features intermediate between that of apo and pro forms of known kallikrein structures. 10858503 0 0 theme mass 83:86 arg1 spectrometry 88:99 tandem mass spectrometry 76:99 tandem mass spectrometry 76:99 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 10858503 4 1 from present 550:556 arg1 % 577:577 approximately 40% 561:577 approximately 40% of the polypeptides 561:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 1 from present 550:556 arg1 polypeptides 586:597 the polypeptides 582:597 the polypeptides 582:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 1 2 theme C-terminally 159:170 arg1 form 182:185 its C-terminally truncated form PRP-3 155:191 its C-terminally truncated form PRP-3 155:191 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 0 3 theme tandem 76:81 arg1 spectrometry 88:99 tandem mass spectrometry 76:99 tandem mass spectrometry 76:99 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 10858503 4 4 from % 577:577 arg1 located 516:522 located 516:522 located 516:522 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 4 from % 577:577 arg1 modification 499:510 The latter modification 488:510 The latter modification 488:510 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 4 from % 577:577 arg1 present 550:556 present 550:556 present 550:556 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 1 5 theme truncated 172:180 arg1 form 182:185 its C-terminally truncated form PRP-3 155:191 its C-terminally truncated form PRP-3 155:191 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 4 6 located present 550:556 arg2 present 550:556 present 550:556 present 550:556 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 6 located present 550:556 arg2 located 516:522 located 516:522 located 516:522 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 6 located present 550:556 arg1 % 577:577 approximately 40% 561:577 approximately 40% of the polypeptides 561:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 6 located present 550:556 arg1 polypeptides 586:597 the polypeptides 582:597 the polypeptides 582:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 6 located present 550:556 arg2 modification 499:510 The latter modification 488:510 The latter modification 488:510 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 0 7 theme proline-rich 40:51 arg1 proteins 53:60 acidic proline-rich proteins 33:60 acidic proline-rich proteins 33:60 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 10858503 3 8 theme glucuronic 444:453 arg1 conjugation 460:470 an O-linked glucuronic acid conjugation 432:470 an O-linked glucuronic acid conjugation 432:470 A pyroglutamic acid residue was demonstrated at the N-terminus, Ser-8 and Ser-22 were shown to be phosphorylated and an O-linked glucuronic acid conjugation was identified. 10858503 0 9 theme novel 2:6 arg1 O-glucuronidation 12:28 A novel Ser O-glucuronidation 0:28 A novel Ser O-glucuronidation in acidic proline-rich proteins 0:60 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 10858503 3 10 theme pyroglutamic 317:328 arg1 residue 335:341 A pyroglutamic acid residue 315:341 A pyroglutamic acid residue 315:341 A pyroglutamic acid residue was demonstrated at the N-terminus, Ser-8 and Ser-22 were shown to be phosphorylated and an O-linked glucuronic acid conjugation was identified. 10858503 1 11 theme Human 102:106 arg1 PRP-1 145:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 0 12 theme Ser 8:10 arg1 O-glucuronidation 12:28 A novel Ser O-glucuronidation 0:28 A novel Ser O-glucuronidation in acidic proline-rich proteins 0:60 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 10858503 3 13 theme acid 330:333 arg1 residue 335:341 A pyroglutamic acid residue 315:341 A pyroglutamic acid residue 315:341 A pyroglutamic acid residue was demonstrated at the N-terminus, Ser-8 and Ser-22 were shown to be phosphorylated and an O-linked glucuronic acid conjugation was identified. 10858503 4 14 theme latter 492:497 arg1 located 516:522 located 516:522 located 516:522 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 14 theme latter 492:497 arg1 modification 499:510 The latter modification 488:510 The latter modification 488:510 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 14 theme latter 492:497 arg1 present 550:556 present 550:556 present 550:556 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 1 15 theme acidic 108:113 arg1 PRP-1 145:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 1 16 theme electrospray 210:221 arg1 spectrometry 235:246 electrospray tandem mass spectrometry 210:246 electrospray tandem mass spectrometry 210:246 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 0 17 theme acidic 33:38 arg1 proteins 53:60 acidic proline-rich proteins 33:60 acidic proline-rich proteins 33:60 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 10858503 4 18 theme polypeptides 586:597 arg1 % 577:577 approximately 40% 561:577 approximately 40% of the polypeptides 561:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 18 theme polypeptides 586:597 arg1 polypeptides 586:597 the polypeptides 582:597 the polypeptides 582:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 1 19 theme tandem 223:228 arg1 spectrometry 235:246 electrospray tandem mass spectrometry 210:246 electrospray tandem mass spectrometry 210:246 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 3 20 theme acid 455:458 arg1 conjugation 460:470 an O-linked glucuronic acid conjugation 432:470 an O-linked glucuronic acid conjugation 432:470 A pyroglutamic acid residue was demonstrated at the N-terminus, Ser-8 and Ser-22 were shown to be phosphorylated and an O-linked glucuronic acid conjugation was identified. 10858503 2 21 theme Post-translational 249:266 arg1 modifications 268:280 Post-translational modifications 249:280 Post-translational modifications 249:280 Post-translational modifications were detected and characterized. 10858503 1 22 theme mass 230:233 arg1 spectrometry 235:246 electrospray tandem mass spectrometry 210:246 electrospray tandem mass spectrometry 210:246 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 1 23 theme proline-rich 115:126 arg1 PRP-1 145:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 3 24 theme O-linked 435:442 arg1 conjugation 460:470 an O-linked glucuronic acid conjugation 432:470 an O-linked glucuronic acid conjugation 432:470 A pyroglutamic acid residue was demonstrated at the N-terminus, Ser-8 and Ser-22 were shown to be phosphorylated and an O-linked glucuronic acid conjugation was identified. 10858503 1 25 theme salivary 128:135 arg1 PRP-1 145:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 3 26 link O-linked 435:442 arg1 conjugation 460:470 an O-linked glucuronic acid conjugation 432:470 an O-linked glucuronic acid conjugation 432:470 A pyroglutamic acid residue was demonstrated at the N-terminus, Ser-8 and Ser-22 were shown to be phosphorylated and an O-linked glucuronic acid conjugation was identified. 10858503 1 27 theme protein 137:143 arg1 PRP-1 145:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 102:149 Human acidic proline-rich salivary protein PRP-1 and its C-terminally truncated form PRP-3 were analyzed by electrospray tandem mass spectrometry. 10858503 4 28 attach present 550:556 arg2 present 550:556 present 550:556 present 550:556 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 28 attach present 550:556 arg1 % 577:577 approximately 40% 561:577 approximately 40% of the polypeptides 561:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 28 attach present 550:556 arg2 modification 499:510 The latter modification 488:510 The latter modification 488:510 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 28 attach present 550:556 arg1 polypeptides 586:597 the polypeptides 582:597 the polypeptides 582:597 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 4 28 attach present 550:556 arg2 located 516:522 located 516:522 located 516:522 The latter modification was located to Ser-17 and found to be present in approximately 40% of the polypeptides. 10858503 0 29 from O-glucuronidation 12:28 arg1 proteins 53:60 acidic proline-rich proteins 33:60 acidic proline-rich proteins 33:60 A novel Ser O-glucuronidation in acidic proline-rich proteins identified by tandem mass spectrometry. 3542030 0 0 theme blood 64:68 arg1 plasma 70:75 human blood plasma 58:75 human blood plasma 58:75 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 5 1 theme Hammond 614:620 arg1 H. 671:672 H. 671:672 H. 671:672 Preparations of Hammond et al. [Hammond, G. L., Robinson, P. A., Sugino, H., Ward, D. N., & Finne, J. (1986) J. Steroid Biochem. 3542030 5 1 theme Hammond 614:620 arg1 [Hammond 629:636 Hammond et al. [Hammond 614:636 Hammond et al. [Hammond 614:636 Preparations of Hammond et al. [Hammond, G. L., Robinson, P. A., Sugino, H., Ward, D. N., & Finne, J. (1986) J. Steroid Biochem. 3542030 10 2 theme sites 1350:1354 arg1 components 1316:1325 possible components 1307:1325 possible components of the steroid-binding sites 1307:1354 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 10 2 theme sites 1350:1354 arg1 segments 1281:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 1 3 theme binding 121:127 arg1 SBP 138:140 SBP 138:140 SBP 138:140 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 1 3 theme binding 121:127 arg1 protein 129:135 sex steroid binding protein 109:135 the sex steroid binding protein (SBP) 105:141 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 8 4 theme disulfide 915:923 arg1 bonds 925:929 The two disulfide bonds 907:929 The two disulfide bonds 907:929 The two disulfide bonds connect Cys-164 to Cys-188 and Cys-333 to Cys-361. 3542030 9 5 theme Asn-Gly 1242:1248 arg1 sequences 1250:1258 two Asn-Gly sequences 1238:1258 two Asn-Gly sequences 1238:1258 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 0 6 theme human 58:62 arg1 plasma 70:75 human blood plasma 58:75 human blood plasma 58:75 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 4 7 theme protein 471:477 arg1 molecules 479:487 the protein molecules 467:487 the protein molecules 467:487 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 10 8 theme hydrophobic 1269:1279 arg1 components 1316:1325 possible components 1307:1325 possible components of the steroid-binding sites 1307:1354 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 10 8 theme hydrophobic 1269:1279 arg1 segments 1281:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 1 9 theme protein 129:135 arg1 sequence 93:100 The amino acid sequence 78:100 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma 78:159 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 4 10 theme other 547:551 arg1 half 553:556 the other half 543:556 the other half 543:556 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 6 11 theme additional 744:753 arg1 leucine 755:761 an additional leucine 741:761 an additional leucine at the amino terminus 741:783 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 10 12 theme Certain 1261:1267 arg1 components 1316:1325 possible components 1307:1325 possible components of the steroid-binding sites 1307:1354 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 10 12 theme Certain 1261:1267 arg1 segments 1281:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 9 13 theme reported 986:993 arg1 heterogeneity 995:1007 The reported heterogeneity 982:1007 The reported heterogeneity of preparations of the molecule 982:1039 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 11 14 theme receptors 1461:1469 arg1 sequences 1416:1424 the cDNA-derived sequences 1399:1424 the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other 1399:1518 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 3 15 theme peptides 356:363 arg1 analysis 344:351 analysis 344:351 analysis of peptides derived by cleavage at either lysyl or methionyl residues 344:421 The sequence was solved primarily by analysis of peptides derived by cleavage at either lysyl or methionyl residues. 3542030 0 16 theme plasma 70:75 arg1 protein 47:53 the sex steroid binding protein 23:53 the sex steroid binding protein of human blood plasma 23:75 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 11 17 theme data 1567:1570 arg1 base 1572:1575 the 1986 data base 1558:1575 the 1986 data base 1558:1575 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 5 18 theme [Hammond 629:636 arg1 Preparations 598:609 Preparations 598:609 Preparations of Hammond et al. [Hammond, G. L., Robinson, P. A., Sugino, H., Ward, D. N., & Finne, J. (1986) J. Steroid Biochem 598:724 Preparations of Hammond et al. [Hammond, G. L., Robinson, P. A., Sugino, H., Ward, D. N., & Finne, J. (1986) J. Steroid Biochem. 3542030 6 19 theme residues 808:815 arg1 total 795:799 a total 793:799 a total of 373 residues in the chain 793:828 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 11 20 link cDNA-derived 1403:1414 arg1 sequences 1416:1424 the cDNA-derived sequences 1399:1424 the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other 1399:1518 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 10 21 theme possible 1307:1314 arg1 components 1316:1325 possible components 1307:1325 possible components of the steroid-binding sites 1307:1354 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 10 21 theme possible 1307:1314 arg1 segments 1281:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments 1261:1288 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 2 22 theme polypeptide 224:234 arg1 chain 236:240 a 373-residue polypeptide chain 210:240 a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains 210:304 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 4 23 dep Arg-Pro-Val-Leu-Pro 522:540 arg1 lack 558:561 lack 558:561 lack Arg-Pro 558:569 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 4 23 dep Arg-Pro-Val-Leu-Pro 522:540 arg1 begin 575:579 begin 575:579 begin with the valine 575:595 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 6 24 from residues 808:815 arg1 chain 824:828 the chain 820:828 the chain 820:828 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 9 25 theme preparations 1012:1023 arg1 heterogeneity 995:1007 The reported heterogeneity 982:1007 The reported heterogeneity of preparations of the molecule 982:1039 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 2 26 theme 373-residue 212:222 arg1 chain 236:240 a 373-residue polypeptide chain 210:240 a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains 210:304 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 11 27 with homology 1378:1385 arg1 sequence 1546:1553 any other protein sequence 1528:1553 any other protein sequence in the 1986 data base 1528:1575 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 11 27 with homology 1378:1385 arg1 sequences 1416:1424 the cDNA-derived sequences 1399:1424 the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other 1399:1518 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 0 28 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the sex steroid binding protein of human blood plasma. 0:76 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 9 29 theme oligosaccharide 1135:1149 arg1 moieties 1151:1158 the oligosaccharide moieties 1131:1158 the oligosaccharide moieties 1131:1158 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 6 30 from chain 824:828 arg1 total 795:799 a total 793:799 a total of 373 residues in the chain 793:828 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 0 31 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the sex steroid binding protein of human blood plasma. 0:76 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 6 32 theme 373 804:806 arg1 residues 808:815 373 residues 804:815 373 residues in the chain 804:828 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 10 33 theme steroid-binding 1334:1348 arg1 sites 1350:1354 the steroid-binding sites 1330:1354 the steroid-binding sites 1330:1354 Certain hydrophobic segments are suggested as possible components of the steroid-binding sites. 3542030 11 34 theme protein 1538:1544 arg1 sequence 1546:1553 any other protein sequence 1528:1553 any other protein sequence in the 1986 data base 1528:1575 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 5 35 dep Hammond 614:620 arg1 al. 625:627 Hammond et al. 614:627 Hammond et al. [Hammond 614:636 Preparations of Hammond et al. [Hammond, G. L., Robinson, P. A., Sugino, H., Ward, D. N., & Finne, J. (1986) J. Steroid Biochem. 3542030 4 36 theme sequence 513:520 arg1 Arg-Pro-Val-Leu-Pro 522:540 the amino-terminal sequence Arg-Pro-Val-Leu-Pro 494:540 the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine 494:595 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 6 37 from total 795:799 arg1 chain 824:828 the chain 820:828 the chain 820:828 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 9 38 theme molecule 1032:1039 arg1 preparations 1012:1023 preparations 1012:1023 preparations of the molecule 1012:1039 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 2 39 theme disulfide 257:265 arg1 bonds 267:271 two disulfide bonds 253:271 two disulfide bonds 253:271 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 1 40 theme human 148:152 arg1 plasma 154:159 human plasma 148:159 human plasma 148:159 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 9 41 gly heterogeneity 995:1007 arg1 preparations 1012:1023 preparations 1012:1023 preparations of the molecule 1012:1039 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 11 42 theme other 1532:1536 arg1 sequence 1546:1553 any other protein sequence 1528:1553 any other protein sequence in the 1986 data base 1528:1575 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 1 43 theme amino 82:86 arg1 sequence 93:100 The amino acid sequence 78:100 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma 78:159 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 0 44 theme sex 27:29 arg1 protein 47:53 the sex steroid binding protein 23:53 the sex steroid binding protein of human blood plasma 23:75 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 2 45 contain containing 242:251 arg1 chain 236:240 a 373-residue polypeptide chain 210:240 a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains 210:304 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 2 45 contain containing 242:251 arg2 bonds 267:271 two disulfide bonds 253:271 two disulfide bonds 253:271 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 2 45 contain containing 242:251 arg2 chains 299:304 three oligosaccharide chains 277:304 three oligosaccharide chains 277:304 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 6 46 contain have 736:739 arg2 leucine 755:761 an additional leucine 741:761 an additional leucine at the amino terminus 741:783 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 6 46 contain have 736:739 arg1 24 727:728 24 727:728 24 727:728 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 2 47 theme SBP 186:188 arg1 subunit 190:196 The SBP subunit 182:196 The SBP subunit 182:196 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 3 48 theme methionyl 404:412 arg1 residues 414:421 methionyl residues 404:421 methionyl residues 404:421 The sequence was solved primarily by analysis of peptides derived by cleavage at either lysyl or methionyl residues. 3542030 7 49 theme Asn 881:883 arg1 residues 885:892 Asn residues 351 and 367 881:904 residues 885:892 Oligosaccharide chains are placed at Thr-7 and at Asn residues 351 and 367. 3542030 9 50 from variations 1117:1126 arg1 part 1107:1110 part 1107:1110 part from variations in the oligosaccharide moieties 1107:1158 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 9 50 from variations 1117:1126 arg1 moieties 1151:1158 the oligosaccharide moieties 1131:1158 the oligosaccharide moieties 1131:1158 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 11 51 theme estrogen 1433:1440 arg1 receptors 1461:1469 the estrogen and glucocorticoid receptors 1429:1469 receptors 1461:1469 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 6 52 dep 24 727:728 arg1 815 731:733 815 731:733 815 731:733 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 0 53 theme binding 39:45 arg1 protein 47:53 the sex steroid binding protein 23:53 the sex steroid binding protein of human blood plasma 23:75 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 4 54 theme molecules 479:487 arg1 half 459:462 approximately half 445:462 approximately half of the protein molecules 445:487 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 6 55 from terminus 776:783 arg1 leucine 755:761 an additional leucine 741:761 an additional leucine at the amino terminus 741:783 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 9 56 from microheterogeneity 1084:1101 arg1 part 1055:1058 part 1055:1058 part from the amino-terminal microheterogeneity 1055:1101 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 11 57 with homologous 1493:1502 arg1 other 1514:1518 other 1514:1518 other 1514:1518 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 3 58 from residues 414:421 arg1 cleavage 376:383 cleavage 376:383 cleavage at either lysyl or methionyl residues 376:421 The sequence was solved primarily by analysis of peptides derived by cleavage at either lysyl or methionyl residues. 3542030 1 59 theme acid 88:91 arg1 sequence 93:100 The amino acid sequence 78:100 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma 78:159 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 0 60 theme steroid 31:37 arg1 protein 47:53 the sex steroid binding protein 23:53 the sex steroid binding protein of human blood plasma 23:75 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 4 61 theme amino-terminal 498:511 arg1 Arg-Pro-Val-Leu-Pro 522:540 the amino-terminal sequence Arg-Pro-Val-Leu-Pro 494:540 the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine 494:595 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 5 62 dep H. 671:672 arg1 Biochem 718:724 Biochem 718:724 Biochem 718:724 Preparations of Hammond et al. [Hammond, G. L., Robinson, P. A., Sugino, H., Ward, D. N., & Finne, J. (1986) J. Steroid Biochem. 3542030 6 63 theme amino 770:774 arg1 terminus 776:783 the amino terminus 766:783 the amino terminus 766:783 24, 815] have an additional leucine at the amino terminus, making a total of 373 residues in the chain. 3542030 11 64 theme cDNA-derived 1403:1414 arg1 sequences 1416:1424 the cDNA-derived sequences 1399:1424 the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other 1399:1518 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 1 65 from plasma 154:159 arg1 sequence 93:100 The amino acid sequence 78:100 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma 78:159 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 2 66 theme oligosaccharide 283:297 arg1 chains 299:304 three oligosaccharide chains 277:304 three oligosaccharide chains 277:304 The SBP subunit consists of a 373-residue polypeptide chain containing two disulfide bonds and three oligosaccharide chains. 3542030 11 67 theme glucocorticoid 1446:1459 arg1 receptors 1461:1469 the estrogen and glucocorticoid receptors 1429:1469 receptors 1461:1469 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 4 68 contain have 489:492 arg2 Arg-Pro-Val-Leu-Pro 522:540 the amino-terminal sequence Arg-Pro-Val-Leu-Pro 494:540 the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine 494:595 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 4 68 contain have 489:492 arg1 half 459:462 approximately half 445:462 approximately half of the protein molecules 445:487 In our preparations, approximately half of the protein molecules have the amino-terminal sequence Arg-Pro-Val-Leu-Pro; the other half lack Arg-Pro and begin with the valine. 3542030 9 69 theme amino-terminal 1069:1082 arg1 microheterogeneity 1084:1101 the amino-terminal microheterogeneity 1065:1101 the amino-terminal microheterogeneity 1065:1101 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 9 70 theme cyclic 1212:1217 arg1 formation 1225:1233 cyclic imide formation 1212:1233 cyclic imide formation 1212:1233 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 11 71 from sequence 1546:1553 arg1 base 1572:1575 the 1986 data base 1558:1575 the 1986 data base 1558:1575 The protein shows no homology either with the cDNA-derived sequences of the estrogen and glucocorticoid receptors found by others to be homologous with each other or with any other protein sequence in the 1986 data base. 3542030 7 72 theme Oligosaccharide 831:845 arg1 chains 847:852 Oligosaccharide chains 831:852 Oligosaccharide chains 831:852 Oligosaccharide chains are placed at Thr-7 and at Asn residues 351 and 367. 3542030 1 73 theme sex 109:111 arg1 SBP 138:140 SBP 138:140 SBP 138:140 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 1 73 theme sex 109:111 arg1 protein 129:135 sex steroid binding protein 109:135 the sex steroid binding protein (SBP) 105:141 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 9 74 theme imide 1219:1223 arg1 formation 1225:1233 cyclic imide formation 1212:1233 cyclic imide formation 1212:1233 The reported heterogeneity of preparations of the molecule may result in part from the amino-terminal microheterogeneity, in part from variations in the oligosaccharide moieties, and possibly in part from rearrangements involving cyclic imide formation in two Asn-Gly sequences. 3542030 0 75 theme protein 47:53 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of the sex steroid binding protein of human blood plasma. 0:76 Amino acid sequence of the sex steroid binding protein of human blood plasma. 3542030 3 76 from lysyl 395:399 arg1 cleavage 376:383 cleavage 376:383 cleavage at either lysyl or methionyl residues 376:421 The sequence was solved primarily by analysis of peptides derived by cleavage at either lysyl or methionyl residues. 3542030 1 77 theme steroid 113:119 arg1 SBP 138:140 SBP 138:140 SBP 138:140 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 3542030 1 77 theme steroid 113:119 arg1 protein 129:135 sex steroid binding protein 109:135 the sex steroid binding protein (SBP) 105:141 The amino acid sequence of the sex steroid binding protein (SBP) from human plasma has been determined. 23010571 2 0 theme NTR 491:493 arg1 module 496:501 a single C-terminal netrin-like (NTR) module 458:501 a single C-terminal netrin-like (NTR) module 458:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 5 1 theme located 775:781 arg1 glycoprotein 762:773 a secreted, N-glycosylated 60kDa glycoprotein 729:773 a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells 729:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 1 theme located 775:781 arg1 ADAMTSL5 717:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 2 dep secreted 731:738 arg1 N-glycosylated 741:754 N-glycosylated 741:754 N-glycosylated 741:754 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 3 theme Recombinant 705:715 arg1 glycoprotein 762:773 a secreted, N-glycosylated 60kDa glycoprotein 729:773 a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells 729:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 3 theme Recombinant 705:715 arg1 ADAMTSL5 717:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 7 4 theme blot 960:963 arg1 analysis 965:972 Western blot analysis 952:972 Western blot analysis 952:972 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 3 5 theme Alternative 504:514 arg1 splicing 516:523 Alternative splicing 504:523 Alternative splicing of ADAMTSL5 at its 5' end 504:549 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 12 6 contain have 1762:1765 arg1 ADAMTSL5 1747:1754 ADAMTSL5 1747:1754 ADAMTSL5 1747:1754 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 12 6 contain have 1762:1765 arg2 role 1769:1772 a role 1767:1772 a role 1767:1772 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 8 7 theme skeletal 1174:1181 arg1 muscle 1183:1188 skeletal muscle 1174:1188 skeletal muscle 1174:1188 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 0 8 theme superfamily 183:193 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 8 theme superfamily 183:193 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 10 9 theme fibroblasts 1479:1489 arg1 matrix 1460:1465 the extracellular matrix 1442:1465 the extracellular matrix of cultured fibroblasts 1442:1489 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 5 10 gly glycoprotein 762:773 arg1 glycoprotein 762:773 a secreted, N-glycosylated 60kDa glycoprotein 729:773 a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells 729:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 10 gly glycoprotein 762:773 arg1 ADAMTSL5 717:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 7 11 theme proteolytic 984:994 arg1 release 996:1002 proteolytic release 984:1002 proteolytic release of the NTR module in transfected cells as well as in some mouse tissues 984:1074 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 1 12 theme ADAMTS 264:269 arg1 metalloproteases 271:286 ADAMTS metalloproteases 264:286 ADAMTS metalloproteases by their similarity to ADAMTS ancillary domains 264:334 ADAMTS-like proteins are related to ADAMTS metalloproteases by their similarity to ADAMTS ancillary domains. 23010571 5 13 theme secreted 731:738 arg1 glycoprotein 762:773 a secreted, N-glycosylated 60kDa glycoprotein 729:773 a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells 729:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 13 theme secreted 731:738 arg1 ADAMTSL5 717:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 6 14 theme blot 890:893 arg1 analysis 895:902 western blot analysis 882:902 western blot analysis 882:902 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 23010571 0 15 theme ADAMTS 176:181 arg1 superfamily 183:193 the ADAMTS superfamily 172:193 the ADAMTS superfamily containing a netrin-like module 172:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 12 16 theme microfibril 1788:1798 arg1 functions 1800:1808 microfibril functions 1788:1808 microfibril functions 1788:1808 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 12 17 theme animal 1729:1734 arg1 mutations 1736:1744 human and animal mutations 1719:1744 human and animal mutations 1719:1744 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 12 18 theme mutations 1736:1744 arg1 identification 1701:1714 identification 1701:1714 identification of human and animal mutations 1701:1744 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 2 19 with superfamily 397:407 arg1 organization 431:442 a unique modular organization 414:442 a unique modular organization that includes a single C-terminal netrin-like (NTR) module 414:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 10 20 theme microfibril 1526:1536 arg1 assembly 1538:1545 microfibril assembly 1526:1545 microfibril assembly 1526:1545 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 0 21 contain containing 46:55 arg2 type 72:75 thrombospondin type 1 motif-like 5 57:90 thrombospondin type 1 motif-like 5 (ADAMTSL5) 57:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 21 contain containing 46:55 arg2 ADAMTSL5 93:100 ADAMTSL5 93:100 ADAMTSL5 93:100 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 21 contain containing 46:55 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 21 contain containing 46:55 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 3 22 theme ADAMTSL5 528:535 arg1 splicing 516:523 Alternative splicing 504:523 Alternative splicing of ADAMTSL5 at its 5' end 504:549 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 7 23 from release 996:1002 arg1 tissues 1068:1074 some mouse tissues 1057:1074 some mouse tissues 1057:1074 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 7 23 from release 996:1002 arg1 cells 1037:1041 transfected cells 1025:1041 transfected cells 1025:1041 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 12 24 theme ADAMTS 1643:1648 arg1 proteins 1650:1657 other ADAMTS proteins 1637:1657 other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations 1637:1744 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 7 25 theme mouse 1062:1066 arg1 tissues 1068:1074 some mouse tissues 1057:1074 some mouse tissues 1057:1074 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 5 26 theme transfected 852:862 arg1 cells 864:868 transfected cells 852:868 transfected cells 852:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 0 27 theme disintegrin-like 2:17 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 27 theme disintegrin-like 2:17 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 8 28 theme musculoskeletal 1142:1156 arg1 tissues 1158:1164 musculoskeletal tissues 1142:1164 musculoskeletal tissues such as skeletal muscle, cartilage and bone 1142:1208 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 8 28 theme musculoskeletal 1142:1156 arg1 bone 1205:1208 bone 1205:1208 bone 1205:1208 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 8 28 theme musculoskeletal 1142:1156 arg1 cartilage 1191:1199 cartilage 1191:1199 cartilage 1191:1199 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 8 28 theme musculoskeletal 1142:1156 arg1 muscle 1183:1188 skeletal muscle 1174:1188 skeletal muscle 1174:1188 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 6 29 theme western 882:888 arg1 analysis 895:902 western blot analysis 882:902 western blot analysis 882:902 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 23010571 5 30 theme cells 864:868 arg1 medium 842:847 the medium 838:847 the medium of transfected cells 838:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 8 31 theme mouse 1099:1103 arg1 organogenesis 1105:1117 mouse organogenesis 1099:1117 mouse organogenesis 1099:1117 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 0 32 theme netrin-like 208:218 arg1 module 220:225 a netrin-like module 206:225 a netrin-like module 206:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 2 33 theme novel 377:381 arg1 ADAMTSL5 365:372 ADAMTSL5 365:372 ADAMTSL5 365:372 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 2 33 theme novel 377:381 arg1 member 383:388 a novel member 375:388 a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module 375:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 0 34 theme metalloprotease 23:37 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 34 theme metalloprotease 23:37 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 35 theme fibrillin-1- 114:125 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 35 theme fibrillin-1- 114:125 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 7 36 theme Western 952:958 arg1 analysis 965:972 Western blot analysis 952:972 Western blot analysis 952:972 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 10 37 theme discernible 1504:1514 arg1 effect 1516:1521 discernible effect 1504:1521 discernible effect on microfibril assembly 1504:1545 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 2 38 theme unique 416:421 arg1 organization 431:442 a unique modular organization 414:442 a unique modular organization that includes a single C-terminal netrin-like (NTR) module 414:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 2 39 theme modular 423:429 arg1 organization 431:442 a unique modular organization 414:442 a unique modular organization that includes a single C-terminal netrin-like (NTR) module 414:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 0 40 theme novel 108:112 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 40 theme novel 108:112 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 3 41 theme mature 629:634 arg1 peptides 606:613 different signal peptides 589:613 different signal peptides 589:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 41 theme mature 629:634 arg1 transcripts 565:575 two transcripts 561:575 two transcripts that encode different signal peptides 561:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 41 theme mature 629:634 arg1 protein 636:642 the same mature protein 620:642 the same mature protein 620:642 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 4 42 theme translational 679:691 arg1 efficiency 693:702 their translational efficiency 673:702 their translational efficiency 673:702 These transcripts differ in their translational efficiency. 23010571 6 43 theme adult 907:911 arg1 tissues 919:925 adult mouse tissues 907:925 adult mouse tissues 907:925 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 23010571 2 44 theme netrin-like 478:488 arg1 module 496:501 a single C-terminal netrin-like (NTR) module 458:501 a single C-terminal netrin-like (NTR) module 458:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 12 45 theme microfibril 1673:1683 arg1 biology 1685:1691 microfibril biology 1673:1691 microfibril biology 1673:1691 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 0 46 theme fibrillin-2- 128:139 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 46 theme fibrillin-2- 128:139 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 47 dep type 72:75 arg1 5 90:90 5 90:90 5 90:90 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 10 48 theme Recombinant 1330:1340 arg1 ADAMTSL5 1342:1349 Recombinant ADAMTSL5 1330:1349 Recombinant ADAMTSL5 1330:1349 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 2 49 theme C-terminal 467:476 arg1 module 496:501 a single C-terminal netrin-like (NTR) module 458:501 a single C-terminal netrin-like (NTR) module 458:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 1 50 theme ADAMTS 311:316 arg1 domains 328:334 ADAMTS ancillary domains 311:334 ADAMTS ancillary domains 311:334 ADAMTS-like proteins are related to ADAMTS metalloproteases by their similarity to ADAMTS ancillary domains. 23010571 5 51 theme subcellular 790:800 arg1 matrix 802:807 the subcellular matrix 786:807 the subcellular matrix 786:807 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 3 52 theme different 589:597 arg1 peptides 606:613 different signal peptides 589:613 different signal peptides 589:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 52 theme different 589:597 arg1 transcripts 565:575 two transcripts 561:575 two transcripts that encode different signal peptides 561:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 52 theme different 589:597 arg1 protein 636:642 the same mature protein 620:642 the same mature protein 620:642 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 7 53 theme module 1015:1020 arg1 release 996:1002 proteolytic release 984:1002 proteolytic release of the NTR module in transfected cells as well as in some mouse tissues 984:1074 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 2 54 theme superfamily 397:407 arg1 ADAMTSL5 365:372 ADAMTSL5 365:372 ADAMTSL5 365:372 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 2 54 theme superfamily 397:407 arg1 member 383:388 a novel member 375:388 a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module 375:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 2 55 theme single 460:465 arg1 module 496:501 a single C-terminal netrin-like (NTR) module 458:501 a single C-terminal netrin-like (NTR) module 458:501 Here, we have characterized ADAMTSL5, a novel member of the superfamily with a unique modular organization that includes a single C-terminal netrin-like (NTR) module. 23010571 1 56 theme ancillary 318:326 arg1 domains 328:334 ADAMTS ancillary domains 311:334 ADAMTS ancillary domains 311:334 ADAMTS-like proteins are related to ADAMTS metalloproteases by their similarity to ADAMTS ancillary domains. 23010571 3 57 theme same 624:627 arg1 peptides 606:613 different signal peptides 589:613 different signal peptides 589:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 57 theme same 624:627 arg1 transcripts 565:575 two transcripts 561:575 two transcripts that encode different signal peptides 561:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 57 theme same 624:627 arg1 protein 636:642 the same mature protein 620:642 the same mature protein 620:642 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 12 58 theme other 1637:1641 arg1 proteins 1650:1657 other ADAMTS proteins 1637:1657 other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations 1637:1744 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 5 59 from medium 842:847 arg1 located 775:781 located 775:781 located 775:781 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 10 60 from microfibrils 1426:1437 arg1 matrix 1460:1465 the extracellular matrix 1442:1465 the extracellular matrix of cultured fibroblasts 1442:1489 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 0 61 theme thrombospondin 57:70 arg1 ADAMTSL5 93:100 ADAMTSL5 93:100 ADAMTSL5 93:100 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 61 theme thrombospondin 57:70 arg1 type 72:75 thrombospondin type 1 motif-like 5 57:90 thrombospondin type 1 motif-like 5 (ADAMTSL5) 57:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 6 62 theme mouse 913:917 arg1 tissues 919:925 adult mouse tissues 907:925 adult mouse tissues 907:925 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 23010571 5 63 from located 775:781 arg1 matrix 802:807 the subcellular matrix 786:807 the subcellular matrix 786:807 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 63 from located 775:781 arg1 medium 842:847 the medium 838:847 the medium of transfected cells 838:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 6 64 theme tissues 919:925 arg1 RT-PCR 871:876 RT-PCR 871:876 RT-PCR 871:876 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 23010571 6 64 theme tissues 919:925 arg1 analysis 895:902 western blot analysis 882:902 western blot analysis 882:902 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 23010571 7 65 theme transfected 1025:1035 arg1 cells 1037:1041 transfected cells 1025:1041 transfected cells 1025:1041 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 10 66 theme cultured 1470:1477 arg1 fibroblasts 1479:1489 cultured fibroblasts 1470:1489 cultured fibroblasts 1470:1489 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 7 67 theme NTR 1011:1013 arg1 module 1015:1020 the NTR module 1007:1020 the NTR module 1007:1020 Western blot analysis suggested proteolytic release of the NTR module in transfected cells as well as in some mouse tissues. 23010571 10 68 from effect 1516:1521 arg1 assembly 1538:1545 microfibril assembly 1526:1545 microfibril assembly 1526:1545 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 0 69 theme heparin-binding 146:160 arg1 domain 39:44 A disintegrin-like and metalloprotease domain 0:44 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) 0:101 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 69 theme heparin-binding 146:160 arg1 member 162:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member 106:167 a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module 106:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 70 contain containing 195:204 arg2 module 220:225 a netrin-like module 206:225 a netrin-like module 206:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 0 70 contain containing 195:204 arg1 superfamily 183:193 the ADAMTS superfamily 172:193 the ADAMTS superfamily containing a netrin-like module 172:225 A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. 23010571 1 71 theme ADAMTS-like 228:238 arg1 proteins 240:247 ADAMTS-like proteins 228:247 ADAMTS-like proteins 228:247 ADAMTS-like proteins are related to ADAMTS metalloproteases by their similarity to ADAMTS ancillary domains. 23010571 5 72 theme 60kDa 756:760 arg1 glycoprotein 762:773 a secreted, N-glycosylated 60kDa glycoprotein 729:773 a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells 729:868 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 5 72 theme 60kDa 756:760 arg1 ADAMTSL5 717:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 705:724 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 9 73 theme ADAMTSL5 1297:1304 arg1 heparin-binding 1278:1292 heparin-binding 1278:1292 heparin-binding 1278:1292 Affinity-chromatography demonstrated heparin-binding of ADAMTSL5 through its NTR-module. 23010571 3 74 theme signal 599:604 arg1 peptides 606:613 different signal peptides 589:613 different signal peptides 589:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 74 theme signal 599:604 arg1 transcripts 565:575 two transcripts 561:575 two transcripts that encode different signal peptides 561:613 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 3 74 theme signal 599:604 arg1 protein 636:642 the same mature protein 620:642 the same mature protein 620:642 Alternative splicing of ADAMTSL5 at its 5' end generates two transcripts that encode different signal peptides, but the same mature protein. 23010571 8 75 theme many 1225:1228 arg1 epithelia 1230:1238 many epithelia 1225:1238 many epithelia 1225:1238 Immunostaining during mouse organogenesis identified ADAMTSL5 in musculoskeletal tissues such as skeletal muscle, cartilage and bone, as well as in many epithelia. 23010571 10 76 theme fibrillin 1416:1424 arg1 microfibrils 1426:1437 fibrillin microfibrils 1416:1437 fibrillin microfibrils in the extracellular matrix of cultured fibroblasts 1416:1489 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 10 77 theme extracellular 1446:1458 arg1 matrix 1460:1465 the extracellular matrix 1442:1465 the extracellular matrix of cultured fibroblasts 1442:1489 Recombinant ADAMTSL5 bound to both fibrillin-1 and fibrillin-2, and co-localized with fibrillin microfibrils in the extracellular matrix of cultured fibroblasts, but without discernible effect on microfibril assembly. 23010571 5 78 from matrix 802:807 arg1 located 775:781 located 775:781 located 775:781 Recombinant ADAMTSL5 is a secreted, N-glycosylated 60kDa glycoprotein located in the subcellular matrix, on the cell-surface, and in the medium of transfected cells. 23010571 12 79 theme human 1719:1723 arg1 mutations 1736:1744 human and animal mutations 1719:1744 human and animal mutations 1719:1744 Like other ADAMTS proteins implicated in microfibril biology through identification of human and animal mutations, ADAMTSL5 could have a role in modulating microfibril functions. 23010571 11 80 theme first 1564:1568 arg1 ADAMTSL5 1548:1555 ADAMTSL5 1548:1555 ADAMTSL5 1548:1555 ADAMTSL5 is the first family member shown to bind both fibrillin-1 and fibrillin-2. 23010571 11 80 theme first 1564:1568 arg1 member 1577:1582 the first family member 1560:1582 the first family member shown to bind both fibrillin-1 and fibrillin-2 1560:1629 ADAMTSL5 is the first family member shown to bind both fibrillin-1 and fibrillin-2. 23010571 11 81 theme family 1570:1575 arg1 ADAMTSL5 1548:1555 ADAMTSL5 1548:1555 ADAMTSL5 1548:1555 ADAMTSL5 is the first family member shown to bind both fibrillin-1 and fibrillin-2. 23010571 11 81 theme family 1570:1575 arg1 member 1577:1582 the first family member 1560:1582 the first family member shown to bind both fibrillin-1 and fibrillin-2 1560:1629 ADAMTSL5 is the first family member shown to bind both fibrillin-1 and fibrillin-2. 23010571 6 82 theme broad 934:938 arg1 expression 940:949 broad expression 934:949 broad expression 934:949 RT-PCR and western blot analysis of adult mouse tissues showed broad expression. 8617200 0 0 theme cysteine 78:85 arg1 repeat 87:92 an eight cysteine repeat 69:92 an eight cysteine repeat of its binding protein LTBP-1 69:122 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 3 1 theme LTBP-1 645:650 arg1 TGF-beta1 610:618 TGF-beta1 610:618 TGF-beta1 610:618 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 3 1 theme LTBP-1 645:650 arg1 fragments 632:640 various fragments 624:640 various fragments of LTBP-1 624:650 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 4 2 theme LTBP-1 842:847 arg1 repeat 832:837 the third 8-Cys repeat 816:837 the third 8-Cys repeat of LTBP-1 816:847 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 6 3 theme LTBP-1 1028:1033 arg1 region 1018:1023 The N-terminal region 1003:1023 The N-terminal region of LTBP-1 consisting of the first 400 amino acids 1003:1073 The N-terminal region of LTBP-1 consisting of the first 400 amino acids was found to associate covalently with the ECM. 8617200 2 4 theme complexes 429:437 arg1 secretion 365:373 the secretion 361:373 the secretion 361:373 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 2 4 theme complexes 429:437 arg1 association 398:408 the association 394:408 the association of latent TGF-beta complexes with the extracellular matrix (ECM) 394:473 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 3 5 theme various 624:630 arg1 fragments 632:640 various fragments 624:640 various fragments of LTBP-1 624:650 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 9 6 theme extracellular 1453:1465 arg1 module 1475:1480 an extracellular protein module 1450:1480 an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins 1450:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 9 6 theme extracellular 1453:1465 arg1 able 1490:1493 able 1490:1493 able 1490:1493 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 3 7 from system 577:582 arg1 cells 597:601 mammalian cells 587:601 mammalian cells 587:601 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 4 8 theme 8-Cys 826:830 arg1 repeat 832:837 the third 8-Cys repeat 816:837 the third 8-Cys repeat of LTBP-1 816:847 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 9 9 theme protein 1467:1473 arg1 module 1475:1480 an extracellular protein module 1450:1480 an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins 1450:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 9 9 theme protein 1467:1473 arg1 able 1490:1493 able 1490:1493 able 1490:1493 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 1 10 theme propeptide 243:252 arg1 complexes 213:221 large latent complexes 200:221 large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein 200:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 4 11 theme third 820:824 arg1 repeat 832:837 the third 8-Cys repeat 816:837 the third 8-Cys repeat of LTBP-1 816:847 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 2 12 dep proteins 323:330 arg1 -3 349:350 -3 349:350 -3 349:350 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 2 12 dep proteins 323:330 arg1 -2 342:343 -2 342:343 -2 342:343 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 2 12 dep proteins 323:330 arg1 LTBPs-1 333:339 LTBPs-1 333:339 LTBPs-1 333:339 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 9 13 theme first 1427:1431 arg1 This 1396:1399 This 1396:1399 This 1396:1399 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 9 13 theme first 1427:1431 arg1 demonstration 1433:1445 the first demonstration 1423:1445 the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins 1423:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 0 14 theme binding 101:107 arg1 LTBP-1 117:122 its binding protein LTBP-1 97:122 its binding protein LTBP-1 97:122 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 6 15 theme amino 1063:1067 arg1 acids 1069:1073 the first 400 amino acids 1049:1073 the first 400 amino acids 1049:1073 The N-terminal region of LTBP-1 consisting of the first 400 amino acids was found to associate covalently with the ECM. 8617200 4 16 theme protein 783:789 arg1 complexes 791:799 the fusion protein complexes 772:799 the fusion protein complexes 772:799 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 8 17 theme disulfide 1291:1299 arg1 bonds 1301:1305 cysteine disulfide bonds 1282:1305 cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein 1282:1372 These interactions are mediated by exchanging cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein during the secretion. 8617200 7 18 theme covalent 1184:1191 arg1 interactions 1222:1233 covalent and specific protein-protein interactions 1184:1233 covalent and specific protein-protein interactions 1184:1233 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 4 19 theme TGF-beta1 887:895 arg1 region 877:882 the LAP region 869:882 the LAP region of TGF-beta1 869:895 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 0 20 theme transforming 32:43 arg1 factor-beta 52:62 the small latent transforming growth factor-beta 15:62 the small latent transforming growth factor-beta 15:62 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 8 21 theme cysteine 1282:1289 arg1 bonds 1301:1305 cysteine disulfide bonds 1282:1305 cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein 1282:1372 These interactions are mediated by exchanging cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein during the secretion. 8617200 6 22 theme N-terminal 1007:1016 arg1 region 1018:1023 The N-terminal region 1003:1023 The N-terminal region of LTBP-1 consisting of the first 400 amino acids 1003:1073 The N-terminal region of LTBP-1 consisting of the first 400 amino acids was found to associate covalently with the ECM. 8617200 9 23 with bonds 1526:1530 arg1 proteins 1557:1564 heterologous ligand proteins 1537:1564 heterologous ligand proteins 1537:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 2 24 with secretion 365:373 arg1 ECM 470:472 ECM 470:472 ECM 470:472 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 2 24 with secretion 365:373 arg1 matrix 462:467 the extracellular matrix 444:467 the extracellular matrix (ECM) 444:473 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 8 25 theme 8-Cys 1324:1328 arg1 repeat 1330:1335 the core 8-Cys repeat 1315:1335 the core 8-Cys repeat 1315:1335 These interactions are mediated by exchanging cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein during the secretion. 8617200 3 26 with level 552:556 arg1 system 577:582 an expression system 563:582 an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs 563:751 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 4 27 theme complexes 791:799 arg1 Immunoblotting 754:767 Immunoblotting 754:767 Immunoblotting of the fusion protein complexes 754:799 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 0 28 theme small 19:23 arg1 factor-beta 52:62 the small latent transforming growth factor-beta 15:62 the small latent transforming growth factor-beta 15:62 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 0 29 theme LTBP-1 117:122 arg1 repeat 87:92 an eight cysteine repeat 69:92 an eight cysteine repeat of its binding protein LTBP-1 69:122 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 9 30 theme heterologous 1537:1548 arg1 proteins 1557:1564 heterologous ligand proteins 1537:1564 heterologous ligand proteins 1537:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 3 31 theme beta1-LAP 495:503 arg1 association 480:490 The association 476:490 The association of beta1-LAP with LTBP-1 476:515 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 1 32 theme most 186:189 arg1 cells 191:195 most cells 186:195 most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein 186:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 0 33 theme protein 109:115 arg1 LTBP-1 117:122 its binding protein LTBP-1 97:122 its binding protein LTBP-1 97:122 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 3 34 theme mammalian 587:595 arg1 cells 597:601 mammalian cells 587:601 mammalian cells 587:601 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 9 35 theme cysteine 1507:1514 arg1 bonds 1526:1530 exchange cysteine disulfide bonds 1498:1530 exchange cysteine disulfide bonds with heterologous ligand proteins 1498:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 7 36 theme LTBP 1165:1168 arg1 repeat 1155:1160 an 8-Cys repeat 1146:1160 an 8-Cys repeat of LTBP 1146:1168 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 7 36 theme LTBP 1165:1168 arg1 capable 1173:1179 capable 1173:1179 capable 1173:1179 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 2 37 theme TGF-beta 420:427 arg1 complexes 429:437 latent TGF-beta complexes 413:437 latent TGF-beta complexes 413:437 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 7 38 theme protein-protein 1206:1220 arg1 interactions 1222:1233 covalent and specific protein-protein interactions 1184:1233 covalent and specific protein-protein interactions 1184:1233 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 4 39 theme LAP 873:875 arg1 region 877:882 the LAP region 869:882 the LAP region of TGF-beta1 869:895 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 9 40 theme disulfide 1516:1524 arg1 bonds 1526:1530 exchange cysteine disulfide bonds 1498:1530 exchange cysteine disulfide bonds with heterologous ligand proteins 1498:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 7 41 theme specific 1197:1204 arg1 interactions 1222:1233 covalent and specific protein-protein interactions 1184:1233 covalent and specific protein-protein interactions 1184:1233 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 0 42 theme latent 25:30 arg1 factor-beta 52:62 the small latent transforming growth factor-beta 15:62 the small latent transforming growth factor-beta 15:62 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 2 43 theme latent 413:418 arg1 complexes 429:437 latent TGF-beta complexes 413:437 latent TGF-beta complexes 413:437 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 3 44 theme molecular 542:550 arg1 level 552:556 the molecular level 538:556 the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs 538:751 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 1 45 theme large 200:204 arg1 complexes 213:221 large latent complexes 200:221 large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein 200:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 0 46 theme factor-beta 52:62 arg1 Association 0:10 Association 0:10 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 0:123 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 1 47 theme binding 278:284 arg1 protein 286:292 a binding protein 276:292 a binding protein 276:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 7 48 theme 8-Cys 1149:1153 arg1 repeat 1155:1160 an 8-Cys repeat 1146:1160 an 8-Cys repeat of LTBP 1146:1168 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 7 48 theme 8-Cys 1149:1153 arg1 capable 1173:1179 capable 1173:1179 capable 1173:1179 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 2 49 theme binding 315:321 arg1 proteins 323:330 The latent TGF-beta binding proteins 295:330 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) 295:351 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 0 50 theme growth 45:50 arg1 factor-beta 52:62 the small latent transforming growth factor-beta 15:62 the small latent transforming growth factor-beta 15:62 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 4 51 theme fusion 776:781 arg1 complexes 791:799 the fusion protein complexes 772:799 the fusion protein complexes 772:799 Immunoblotting of the fusion protein complexes indicated that the third 8-Cys repeat of LTBP-1 bound covalently to the LAP region of TGF-beta1. 8617200 3 52 dep the 688:690 arg1 aid 692:694 aid 692:694 aid 692:694 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 2 53 theme TGF-beta 306:313 arg1 proteins 323:330 The latent TGF-beta binding proteins 295:330 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) 295:351 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 3 54 theme signal 701:706 arg1 peptide 708:714 a signal peptide 699:714 a signal peptide synthesized to the LTBP-1 constructs 699:751 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 9 55 theme ligand 1550:1555 arg1 proteins 1557:1564 heterologous ligand proteins 1537:1564 heterologous ligand proteins 1537:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 8 56 theme associated 1355:1364 arg1 protein 1366:1372 an optionally associated protein 1341:1372 an optionally associated protein 1341:1372 These interactions are mediated by exchanging cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein during the secretion. 8617200 0 57 with Association 0:10 arg1 repeat 87:92 an eight cysteine repeat 69:92 an eight cysteine repeat of its binding protein LTBP-1 69:122 Association of the small latent transforming growth factor-beta with an eight cysteine repeat of its binding protein LTBP-1. 8617200 2 58 theme latent 299:304 arg1 proteins 323:330 The latent TGF-beta binding proteins 295:330 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) 295:351 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 8 59 theme core 1319:1322 arg1 repeat 1330:1335 the core 8-Cys repeat 1315:1335 the core 8-Cys repeat 1315:1335 These interactions are mediated by exchanging cysteine disulfide bonds between the core 8-Cys repeat and an optionally associated protein during the secretion. 8617200 1 60 theme latent 206:211 arg1 complexes 213:221 large latent complexes 200:221 large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein 200:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 1 61 from cells 191:195 arg1 complexes 213:221 large latent complexes 200:221 large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein 200:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 3 62 theme expression 566:575 arg1 system 577:582 an expression system 563:582 an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs 563:751 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 1 63 theme Transforming 125:136 arg1 factor-betas 145:156 Transforming growth factor-betas 125:156 Transforming growth factor-betas (TGF-betas) 125:168 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 1 63 theme Transforming 125:136 arg1 TGF-betas 159:167 TGF-betas 159:167 TGF-betas 159:167 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 6 64 theme first 1053:1057 arg1 acids 1069:1073 the first 400 amino acids 1049:1073 the first 400 amino acids 1049:1073 The N-terminal region of LTBP-1 consisting of the first 400 amino acids was found to associate covalently with the ECM. 8617200 2 65 theme extracellular 448:460 arg1 ECM 470:472 ECM 470:472 ECM 470:472 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 2 65 theme extracellular 448:460 arg1 matrix 462:467 the extracellular matrix 444:467 the extracellular matrix (ECM) 444:473 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 5 66 theme beta1-LAP 992:1000 arg1 Cys33 983:987 Cys33 983:987 Cys33 of beta1-LAP 983:1000 The cysteine required for the association between LTBP-1 and beta1-LAP was mapped to Cys33 of beta1-LAP. 8617200 2 67 with association 398:408 arg1 ECM 470:472 ECM 470:472 ECM 470:472 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 2 67 with association 398:408 arg1 matrix 462:467 the extracellular matrix 444:467 the extracellular matrix (ECM) 444:473 The latent TGF-beta binding proteins (LTBPs-1, -2 and -3) mediate the secretion and, subsequently, the association of latent TGF-beta complexes with the extracellular matrix (ECM). 8617200 1 68 theme growth 138:143 arg1 factor-betas 145:156 Transforming growth factor-betas 125:156 Transforming growth factor-betas (TGF-betas) 125:168 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 1 68 theme growth 138:143 arg1 TGF-betas 159:167 TGF-betas 159:167 TGF-betas 159:167 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8617200 7 69 theme interactions 1222:1233 arg1 repeat 1155:1160 an 8-Cys repeat 1146:1160 an 8-Cys repeat of LTBP 1146:1168 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 7 69 theme interactions 1222:1233 arg1 capable 1173:1179 capable 1173:1179 capable 1173:1179 The data indicate that an 8-Cys repeat of LTBP is capable of covalent and specific protein-protein interactions. 8617200 9 70 theme module 1475:1480 arg1 This 1396:1399 This 1396:1399 This 1396:1399 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 9 70 theme module 1475:1480 arg1 demonstration 1433:1445 the first demonstration 1423:1445 the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins 1423:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 3 71 with association 480:490 arg1 LTBP-1 510:515 LTBP-1 510:515 LTBP-1 510:515 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 9 72 theme exchange 1498:1505 arg1 bonds 1526:1530 exchange cysteine disulfide bonds 1498:1530 exchange cysteine disulfide bonds with heterologous ligand proteins 1498:1564 This is, to our knowledge, the first demonstration of an extracellular protein module that is able to exchange cysteine disulfide bonds with heterologous ligand proteins. 8617200 3 73 theme LTBP-1 735:740 arg1 constructs 742:751 the LTBP-1 constructs 731:751 the LTBP-1 constructs 731:751 The association of beta1-LAP with LTBP-1 was characterized at the molecular level with an expression system in mammalian cells, where TGF-beta1 and various fragments of LTBP-1 were co-expressed and secreted with the aid of a signal peptide synthesized to the LTBP-1 constructs. 8617200 1 74 theme TGF-beta 226:233 arg1 complexes 213:221 large latent complexes 200:221 large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein 200:292 Transforming growth factor-betas (TGF-betas) are produced by most cells in large latent complexes of TGF-beta and its propeptide (LAP) associated with a binding protein. 8069634 0 0 theme moiety 151:156 arg1 role 132:135 role 132:135 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 0:157 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 13 1 theme large 2025:2029 arg1 area 2053:2056 a large complementary surface area 2023:2056 a large complementary surface area between the oligosaccharide and the lectin 2023:2099 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 13 2 theme mammalian 2222:2230 arg1 lectin 2232:2237 a C-type mammalian lectin 2213:2237 a C-type mammalian lectin 2213:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 6 3 theme sugar 1205:1209 arg1 conformation 1211:1222 the sugar conformation 1201:1222 the sugar conformation 1201:1222 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 4 4 theme lectins 630:636 arg1 structures 608:617 the structures 604:617 the structures of several lectins complexed with incomplete oligosaccharides 604:679 While the structures of several lectins complexed with incomplete oligosaccharides have been solved, no previous structure has included the complete glycoprotein. 8069634 10 5 theme concanavalin 1626:1637 arg1 ConA 1642:1645 ConA 1642:1645 ConA 1642:1645 Comparison of our structure with that of concanavalin A (ConA) suggests that the ConA binding site cannot accommodate this fucose. 8069634 10 5 theme concanavalin 1626:1637 arg1 A 1639:1639 concanavalin A 1626:1639 concanavalin A (ConA) 1626:1646 Comparison of our structure with that of concanavalin A (ConA) suggests that the ConA binding site cannot accommodate this fucose. 8069634 13 6 theme surface 2045:2051 arg1 area 2053:2056 a large complementary surface area 2023:2056 a large complementary surface area between the oligosaccharide and the lectin 2023:2099 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 11 7 theme oligosaccharides 1917:1932 arg1 types 1908:1912 these types 1902:1912 these types of oligosaccharides 1902:1932 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 11 8 dep have 1784:1787 arg1 whereas 1869:1875 whereas 1869:1875 whereas 1869:1875 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 4 9 theme previous 702:709 arg1 structure 711:719 no previous structure 699:719 no previous structure 699:719 While the structures of several lectins complexed with incomplete oligosaccharides have been solved, no previous structure has included the complete glycoprotein. 8069634 2 10 from process 390:396 arg1 cycle 414:418 the nitrogen cycle 401:418 the nitrogen cycle 401:418 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 11 11 theme unfucosylated 1849:1861 arg1 ones 1863:1866 unfucosylated ones 1849:1866 unfucosylated ones 1849:1866 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 0 12 theme isolated 97:104 arg1 glycopeptide 118:129 an isolated biantennary glycopeptide 94:129 an isolated biantennary glycopeptide 94:129 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 13 13 theme recognition 2191:2201 arg1 lectin 2232:2237 a C-type mammalian lectin 2213:2237 a C-type mammalian lectin 2213:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 13 13 theme recognition 2191:2201 arg1 domain 2203:2208 the carbohydrate recognition domain 2174:2208 the carbohydrate recognition domain of a C-type mammalian lectin 2174:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 2 14 theme symbiotic 303:311 arg1 interactions 313:324 the symbiotic interactions 299:324 the symbiotic interactions between plants and nitrogen-fixing microorganisms 299:374 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 5 15 theme lactotransferrin 900:915 arg1 fragment 965:972 an isolated glycopeptide (2.1 kDa) fragment 930:972 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 15 theme lactotransferrin 900:915 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 8 16 theme essential 1310:1318 arg1 moiety 1328:1333 the essential mannose moiety 1306:1333 the essential mannose moiety of the monosaccharide-binding site 1306:1368 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 10 17 theme ConA 1666:1669 arg1 site 1679:1682 the ConA binding site 1662:1682 the ConA binding site 1662:1682 Comparison of our structure with that of concanavalin A (ConA) suggests that the ConA binding site cannot accommodate this fucose. 8069634 3 18 located found 553:557 arg1 majority 566:573 the majority 562:573 the majority of N-glycosylproteins 562:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 3 18 located found 553:557 arg2 glycans 529:535 the fucosylated biantennary N-acetyllactosamine-type glycans 476:535 the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 476:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 5 19 gly glycopeptide 942:953 arg2 kDa 960:962 2.1 kDa 956:962 2.1 kDa 956:962 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 19 gly glycopeptide 942:953 arg2 glycopeptide 942:953 glycopeptide 942:953 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 13 20 theme complex 2158:2164 arg1 structure 2143:2151 the recently determined structure 2119:2151 the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin 2119:2339 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 4 21 gly glycoprotein 747:758 arg1 glycoprotein 747:758 the complete glycoprotein 734:758 the complete glycoprotein 734:758 While the structures of several lectins complexed with incomplete oligosaccharides have been solved, no previous structure has included the complete glycoprotein. 8069634 9 22 theme isolectin 1549:1557 arg1 complex 1576:1582 the previously determined isolectin I-octasaccharide complex 1523:1582 the previously determined isolectin I-octasaccharide complex 1523:1582 This oligosaccharide conformation differs substantially from that seen in the previously determined isolectin I-octasaccharide complex. 8069634 11 23 contain have 1784:1787 arg1 lectins 1776:1782 Viciae lectins 1769:1782 Viciae lectins 1769:1782 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 11 23 contain have 1784:1787 arg2 affinity 1798:1805 a higher affinity 1789:1805 a higher affinity for fucosylated oligosaccharides 1789:1838 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 0 24 gly glycopeptide 118:129 arg2 glycopeptide 118:129 an isolated biantennary glycopeptide 94:129 an isolated biantennary glycopeptide 94:129 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 6 25 contain has 1134:1136 arg2 influence 1145:1153 little influence 1138:1153 little influence 1138:1153 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 6 25 contain has 1134:1136 arg1 part 1109:1112 the protein part 1097:1112 the protein part of the glycoprotein 1097:1132 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 8 26 theme site 1365:1368 arg1 moiety 1328:1333 the essential mannose moiety 1306:1333 the essential mannose moiety of the monosaccharide-binding site 1306:1368 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 5 27 theme Lathyrus 814:821 arg1 isolectin 830:838 Lathyrus ochrus isolectin II 814:841 Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 814:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 13 28 theme determined 2132:2141 arg1 structure 2143:2151 the recently determined structure 2119:2151 the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin 2119:2339 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 5 29 theme glycopeptide 942:953 arg1 fragment 965:972 an isolated glycopeptide (2.1 kDa) fragment 930:972 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 6 30 gly glycoprotein 1121:1132 arg1 glycoprotein 1121:1132 the glycoprotein 1117:1132 the glycoprotein 1117:1132 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 0 31 theme N2 72:73 arg1 fragment 75:82 the human lactotransferrin N2 fragment 45:82 the human lactotransferrin N2 fragment 45:82 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 10 32 with Comparison 1585:1594 arg1 that 1618:1621 that 1618:1621 that 1618:1621 Comparison of our structure with that of concanavalin A (ConA) suggests that the ConA binding site cannot accommodate this fucose. 8069634 11 33 theme fucosylated 1811:1821 arg1 oligosaccharides 1823:1838 fucosylated oligosaccharides 1811:1838 fucosylated oligosaccharides 1811:1838 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 5 34 from fragment 882:889 arg1 A 1008:1008 3.3 A and 2.8 A resolution 1004:1029 A 1008:1008 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 34 from fragment 882:889 arg1 resolution 1020:1029 3.3 A and 2.8 A resolution 1004:1029 resolution 1020:1029 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 13 35 with contrast 2105:2112 arg1 structure 2143:2151 the recently determined structure 2119:2151 the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin 2119:2339 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 5 36 from fragment 965:972 arg1 A 1008:1008 3.3 A and 2.8 A resolution 1004:1029 A 1008:1008 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 36 from fragment 965:972 arg1 resolution 1020:1029 3.3 A and 2.8 A resolution 1004:1029 resolution 1020:1029 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 6 37 theme protein 1101:1107 arg1 part 1109:1112 the protein part 1097:1112 the protein part of the glycoprotein 1097:1132 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 2 38 theme nitrogen 405:412 arg1 cycle 414:418 the nitrogen cycle 401:418 the nitrogen cycle 401:418 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 5 39 theme monoglycosylated 865:880 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 13 40 theme mannose 2299:2305 arg1 residue 2307:2313 only the non-reducing terminal mannose residue 2268:2313 only the non-reducing terminal mannose residue 2268:2313 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 5 41 theme lactotransferrin 983:998 arg1 fragment 965:972 an isolated glycopeptide (2.1 kDa) fragment 930:972 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 41 theme lactotransferrin 983:998 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 2 42 theme Legume 261:266 arg1 lectins 268:274 Legume lectins 261:274 Legume lectins 261:274 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 2 42 theme Legume 261:266 arg1 mediators 285:293 mediators 285:293 mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms 285:374 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 8 43 theme large 1403:1407 arg1 surface 1409:1415 a large surface 1401:1415 a large surface of interaction with the lectin 1401:1446 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 3 44 gly fucosylated 480:490 arg1 glycans 529:535 the fucosylated biantennary N-acetyllactosamine-type glycans 476:535 the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 476:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 5 45 theme isolated 933:940 arg1 fragment 965:972 an isolated glycopeptide (2.1 kDa) fragment 930:972 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 46 gly monoglycosylated 865:880 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 0 47 theme lectin 23:28 arg1 Structures 0:9 Structures 0:9 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 0:157 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 0 48 theme human 49:53 arg1 fragment 75:82 the human lactotransferrin N2 fragment 45:82 the human lactotransferrin N2 fragment 45:82 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 11 49 dep CONCLUSIONS 1716:1726 arg1 explain 1740:1746 explain 1740:1746 explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 1740:1944 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 3 50 theme biantennary 492:502 arg1 glycans 529:535 the fucosylated biantennary N-acetyllactosamine-type glycans 476:535 the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 476:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 6 51 theme little 1138:1143 arg1 influence 1145:1153 little influence 1138:1153 little influence 1138:1153 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 11 52 theme Viciae 1769:1774 arg1 lectins 1776:1782 Viciae lectins 1769:1782 Viciae lectins 1769:1782 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 1 53 theme oligosaccharide 237:251 arg1 chains 253:258 oligosaccharide chains 237:258 oligosaccharide chains 237:258 BACKGROUND Lectins mediate cell-cell interactions by specifically recognizing oligosaccharide chains. 8069634 0 54 dep Structures 0:9 arg1 role 132:135 role 132:135 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 0:157 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 6 55 theme glycoprotein 1121:1132 arg1 part 1109:1112 the protein part 1097:1112 the protein part of the glycoprotein 1097:1132 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 4 56 theme incomplete 653:662 arg1 oligosaccharides 664:679 incomplete oligosaccharides 653:679 incomplete oligosaccharides 653:679 While the structures of several lectins complexed with incomplete oligosaccharides have been solved, no previous structure has included the complete glycoprotein. 8069634 2 57 theme important 380:388 arg1 process 390:396 an important process 377:396 an important process in the nitrogen cycle 377:418 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 2 57 theme important 380:388 arg1 mediators 285:293 mediators 285:293 mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms 285:374 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 13 58 theme C-type 2215:2220 arg1 lectin 2232:2237 a C-type mammalian lectin 2213:2237 a C-type mammalian lectin 2213:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 7 59 theme same 1270:1273 arg1 conformation 1284:1295 the same extended conformation 1266:1295 the same extended conformation 1266:1295 In both cases the oligosaccharide adopts the same extended conformation. 8069634 13 60 theme complementary 2031:2043 arg1 area 2053:2056 a large complementary surface area 2023:2056 a large complementary surface area between the oligosaccharide and the lectin 2023:2099 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 5 61 from A 1008:1008 arg1 fragment 965:972 an isolated glycopeptide (2.1 kDa) fragment 930:972 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 61 from A 1008:1008 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 61 from A 1008:1008 arg1 lactotransferrin 983:998 human lactotransferrin 977:998 human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 977:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 13 62 theme lectin 2232:2237 arg1 lectin 2232:2237 a C-type mammalian lectin 2213:2237 a C-type mammalian lectin 2213:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 13 62 theme lectin 2232:2237 arg1 domain 2203:2208 the carbohydrate recognition domain 2174:2208 the carbohydrate recognition domain of a C-type mammalian lectin 2174:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 13 62 theme lectin 2232:2237 arg1 oligomannoside 2246:2259 an oligomannoside 2243:2259 an oligomannoside 2243:2259 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 4 63 theme several 622:628 arg1 lectins 630:636 several lectins 622:636 several lectins complexed with incomplete oligosaccharides 622:679 While the structures of several lectins complexed with incomplete oligosaccharides have been solved, no previous structure has included the complete glycoprotein. 8069634 9 64 theme oligosaccharide 1454:1468 arg1 conformation 1470:1481 This oligosaccharide conformation 1449:1481 This oligosaccharide conformation 1449:1481 This oligosaccharide conformation differs substantially from that seen in the previously determined isolectin I-octasaccharide complex. 8069634 6 65 theme complex 1190:1196 arg1 conformation 1211:1222 the sugar conformation 1201:1222 the sugar conformation 1201:1222 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 6 65 theme complex 1190:1196 arg1 stabilization 1169:1181 the stabilization 1165:1181 the stabilization of the complex 1165:1196 Comparison between the two structures showed that the protein part of the glycoprotein has little influence on either the stabilization of the complex or the sugar conformation. 8069634 13 66 theme non-reducing 2277:2288 arg1 residue 2307:2313 only the non-reducing terminal mannose residue 2268:2313 only the non-reducing terminal mannose residue 2268:2313 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 5 67 theme isolectin 830:838 arg1 structures 800:809 the crystal structures 788:809 the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 788:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 68 from resolution 1020:1029 arg1 fragment 965:972 an isolated glycopeptide (2.1 kDa) fragment 930:972 an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 930:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 68 from resolution 1020:1029 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 68 from resolution 1020:1029 arg1 lactotransferrin 983:998 human lactotransferrin 977:998 human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 977:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 1 69 theme BACKGROUND 159:168 arg1 Lectins 170:176 BACKGROUND Lectins 159:176 BACKGROUND Lectins 159:176 BACKGROUND Lectins mediate cell-cell interactions by specifically recognizing oligosaccharide chains. 8069634 8 70 with interaction 1420:1430 arg1 lectin 1441:1446 the lectin 1437:1446 the lectin 1437:1446 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 0 71 theme biantennary 106:116 arg1 glycopeptide 118:129 an isolated biantennary glycopeptide 94:129 an isolated biantennary glycopeptide 94:129 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 3 72 theme N-glycosylproteins 578:595 arg1 majority 566:573 the majority 562:573 the majority of N-glycosylproteins 562:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 5 73 theme human 894:898 arg1 lactotransferrin 900:915 human lactotransferrin 894:915 human lactotransferrin (18 kDa) 894:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 73 theme human 894:898 arg1 kDa 921:923 18 kDa 918:923 18 kDa 918:923 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 10 74 theme binding 1671:1677 arg1 site 1679:1682 the ConA binding site 1662:1682 the ConA binding site 1662:1682 Comparison of our structure with that of concanavalin A (ConA) suggests that the ConA binding site cannot accommodate this fucose. 8069634 11 75 theme ConA 1893:1896 arg1 affinity 1881:1888 the affinity 1877:1888 the affinity of ConA for these types of oligosaccharides 1877:1932 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 11 75 theme ConA 1893:1896 arg1 similar 1937:1943 similar 1937:1943 similar 1937:1943 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 3 76 contain have 451:454 arg2 affinity 463:470 a high affinity 456:470 a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 456:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 3 76 contain have 451:454 arg1 Lectins 421:427 Lectins 421:427 Lectins from the Viciae tribe 421:449 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 8 77 theme mannose 1320:1326 arg1 moiety 1328:1333 the essential mannose moiety 1306:1333 the essential mannose moiety of the monosaccharide-binding site 1306:1368 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 0 78 theme fucose 144:149 arg1 moiety 151:156 the fucose moiety 140:156 the fucose moiety 140:156 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 9 79 theme determined 1538:1547 arg1 complex 1576:1582 the previously determined isolectin I-octasaccharide complex 1523:1582 the previously determined isolectin I-octasaccharide complex 1523:1582 This oligosaccharide conformation differs substantially from that seen in the previously determined isolectin I-octasaccharide complex. 8069634 5 80 dep A 1008:1008 arg1 respectively 1032:1043 respectively 1032:1043 respectively 1032:1043 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 2 81 theme nitrogen-fixing 345:359 arg1 microorganisms 361:374 nitrogen-fixing microorganisms 345:374 nitrogen-fixing microorganisms 345:374 Legume lectins serve as mediators for the symbiotic interactions between plants and nitrogen-fixing microorganisms, an important process in the nitrogen cycle. 8069634 0 82 theme lactotransferrin 55:70 arg1 fragment 75:82 the human lactotransferrin N2 fragment 45:82 the human lactotransferrin N2 fragment 45:82 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 13 83 theme carbohydrate 2178:2189 arg1 lectin 2232:2237 a C-type mammalian lectin 2213:2237 a C-type mammalian lectin 2213:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 13 83 theme carbohydrate 2178:2189 arg1 domain 2203:2208 the carbohydrate recognition domain 2174:2208 the carbohydrate recognition domain of a C-type mammalian lectin 2174:2237 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 8 84 contain has 1397:1399 arg2 surface 1409:1415 a large surface 1401:1415 a large surface of interaction with the lectin 1401:1446 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 8 84 contain has 1397:1399 arg1 fucose-1 1375:1382 the fucose-1 1371:1382 the fucose-1' of the core 1371:1395 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 9 85 theme I-octasaccharide 1559:1574 arg1 complex 1576:1582 the previously determined isolectin I-octasaccharide complex 1523:1582 the previously determined isolectin I-octasaccharide complex 1523:1582 This oligosaccharide conformation differs substantially from that seen in the previously determined isolectin I-octasaccharide complex. 8069634 8 86 theme monosaccharide-binding 1342:1363 arg1 site 1365:1368 the monosaccharide-binding site 1338:1368 the monosaccharide-binding site 1338:1368 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 11 87 theme higher 1791:1796 arg1 affinity 1798:1805 a higher affinity 1789:1805 a higher affinity for fucosylated oligosaccharides 1789:1838 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 5 88 theme ochrus 823:828 arg1 isolectin 830:838 Lathyrus ochrus isolectin II 814:841 Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 814:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 12 89 theme mutagenesis 1978:1988 arg1 experiments 1990:2000 mutagenesis experiments 1978:2000 mutagenesis experiments 1978:2000 This explanation is testable by mutagenesis experiments. 8069634 5 90 dep RESULTS 761:767 arg1 determined 777:786 determined 777:786 have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 772:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 4 91 theme complete 738:745 arg1 glycoprotein 747:758 the complete glycoprotein 734:758 the complete glycoprotein 734:758 While the structures of several lectins complexed with incomplete oligosaccharides have been solved, no previous structure has included the complete glycoprotein. 8069634 5 92 theme crystal 792:798 arg1 structures 800:809 the crystal structures 788:809 the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 788:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 1 93 theme cell-cell 186:194 arg1 interactions 196:207 cell-cell interactions 186:207 cell-cell interactions 186:207 BACKGROUND Lectins mediate cell-cell interactions by specifically recognizing oligosaccharide chains. 8069634 11 94 gly fucosylated 1811:1821 arg1 oligosaccharides 1823:1838 fucosylated oligosaccharides 1811:1838 fucosylated oligosaccharides 1811:1838 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 8069634 3 95 theme high 458:461 arg1 affinity 463:470 a high affinity 456:470 a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 456:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 5 96 theme N2 862:863 arg1 fragment 882:889 the N2 monoglycosylated fragment 858:889 the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) 858:924 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 5 97 theme human 977:981 arg1 lactotransferrin 983:998 human lactotransferrin 977:998 human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively) 977:1044 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 8 98 theme core 1392:1395 arg1 fucose-1 1375:1382 the fucose-1 1371:1382 the fucose-1' of the core 1371:1395 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 0 99 theme legume 16:21 arg1 lectin 23:28 a legume lectin 14:28 a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide 14:129 Structures of a legume lectin complexed with the human lactotransferrin N2 fragment, and with an isolated biantennary glycopeptide: role of the fucose moiety. 8069634 3 100 theme fucosylated 480:490 arg1 glycans 529:535 the fucosylated biantennary N-acetyllactosamine-type glycans 476:535 the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 476:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 3 101 theme Viciae 438:443 arg1 tribe 445:449 the Viciae tribe 434:449 the Viciae tribe 434:449 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 8 102 theme interaction 1420:1430 arg1 surface 1409:1415 a large surface 1401:1415 a large surface of interaction with the lectin 1401:1446 Besides the essential mannose moiety of the monosaccharide-binding site, the fucose-1' of the core has a large surface of interaction with the lectin. 8069634 13 103 theme terminal 2290:2297 arg1 residue 2307:2313 only the non-reducing terminal mannose residue 2268:2313 only the non-reducing terminal mannose residue 2268:2313 Our structure shows a large complementary surface area between the oligosaccharide and the lectin, in contrast with the recently determined structure of a complex between the carbohydrate recognition domain of a C-type mammalian lectin and an oligomannoside, where only the non-reducing terminal mannose residue interacts with the lectin. 8069634 5 104 theme A 1018:1018 arg1 resolution 1020:1029 3.3 A and 2.8 A resolution 1004:1029 resolution 1020:1029 RESULTS We have determined the crystal structures of Lathyrus ochrus isolectin II complexed with the N2 monoglycosylated fragment of human lactotransferrin (18 kDa) and an isolated glycopeptide (2.1 kDa) fragment of human lactotransferrin (at 3.3 A and 2.8 A resolution, respectively). 8069634 3 105 from tribe 445:449 arg1 Lectins 421:427 Lectins 421:427 Lectins from the Viciae tribe 421:449 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 3 106 theme N-acetyllactosamine-type 504:527 arg1 glycans 529:535 the fucosylated biantennary N-acetyllactosamine-type glycans 476:535 the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins 476:595 Lectins from the Viciae tribe have a high affinity for the fucosylated biantennary N-acetyllactosamine-type glycans which are to be found in the majority of N-glycosylproteins. 8069634 7 107 theme extended 1275:1282 arg1 conformation 1284:1295 the same extended conformation 1266:1295 the same extended conformation 1266:1295 In both cases the oligosaccharide adopts the same extended conformation. 8069634 10 108 theme structure 1603:1611 arg1 Comparison 1585:1594 Comparison 1585:1594 Comparison of our structure with that of concanavalin A (ConA) 1585:1646 Comparison of our structure with that of concanavalin A (ConA) suggests that the ConA binding site cannot accommodate this fucose. 8069634 11 109 gly unfucosylated 1849:1861 arg1 ones 1863:1866 unfucosylated ones 1849:1866 unfucosylated ones 1849:1866 CONCLUSIONS Our results explain the observation that Viciae lectins have a higher affinity for fucosylated oligosaccharides than for unfucosylated ones, whereas the affinity of ConA for these types of oligosaccharides is similar. 10462545 8 0 theme probenecid-sensitive 1226:1245 arg1 uptake 1247:1252 saturable, probenecid-sensitive uptake 1215:1252 uptake 1247:1252 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 5 1 theme skeletal 764:771 arg1 muscle 773:778 skeletal muscle 764:778 skeletal muscle 764:778 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 5 2 from expression 734:743 arg1 brain 754:758 brain 754:758 brain 754:758 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 5 2 from expression 734:743 arg1 muscle 773:778 skeletal muscle 764:778 skeletal muscle 764:778 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 5 3 theme hOAT1 728:732 arg1 expression 734:743 hOAT1 expression 728:743 hOAT1 expression in human brain and skeletal muscle 728:778 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 3 4 attach isolated 473:480 arg2 clone 463:467 A cDNA clone 456:467 A cDNA clone 456:467 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 3 4 attach isolated 473:480 arg1 library 501:507 a human renal library 487:507 a human renal library 487:507 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 1 5 theme dose-limiting 152:164 arg1 effect 183:188 the dose-limiting clinical adverse effect 148:188 the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics 148:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 5 theme dose-limiting 152:164 arg1 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 4 6 theme Northern 567:574 arg1 analysis 576:583 Northern analysis 567:583 Northern analysis 567:583 Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. 10462545 5 7 theme human 748:752 arg1 brain 754:758 brain 754:758 brain 754:758 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 6 8 theme cortex 826:831 arg1 analysis 801:808 Immunoblot analysis 790:808 Immunoblot analysis of human kidney cortex 790:831 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 10 9 theme active 1654:1659 arg1 role 1661:1664 an active role 1651:1664 an active role 1651:1664 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 9 10 theme 238 1468:1470 arg1 microM 1480:1485 238 and 270 microM 1468:1485 microM 1480:1485 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 9 11 theme 270 1476:1478 arg1 microM 1480:1485 238 and 270 microM 1468:1485 microM 1480:1485 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 6 12 theme abundant 918:925 arg1 N-glycosylation 927:941 abundant N-glycosylation 918:941 abundant N-glycosylation 918:941 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 6 13 theme kidney 819:824 arg1 cortex 826:831 human kidney cortex 813:831 human kidney cortex 813:831 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 10 14 theme proximal 1618:1625 arg1 tubules 1627:1633 renal proximal tubules 1612:1633 renal proximal tubules 1612:1633 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 8 15 theme anion 1177:1181 arg1 transporter 1183:1193 rat renal organic anion transporter 1 1159:1195 rat renal organic anion transporter 1 (rROAT1) 1159:1204 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 8 15 theme anion 1177:1181 arg1 rROAT1 1198:1203 rROAT1 1198:1203 rROAT1 1198:1203 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 0 16 theme organic 101:107 arg1 transporter 115:125 renal organic anion transporter 1 95:127 renal organic anion transporter 1 95:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 3 17 theme human 489:493 arg1 library 501:507 a human renal library 487:507 a human renal library 487:507 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 8 18 theme renal 1163:1167 arg1 transporter 1183:1193 rat renal organic anion transporter 1 1159:1195 rat renal organic anion transporter 1 (rROAT1) 1159:1204 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 8 18 theme renal 1163:1167 arg1 rROAT1 1198:1203 rROAT1 1198:1203 rROAT1 1198:1203 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 6 19 theme human 813:817 arg1 cortex 826:831 human kidney cortex 813:831 human kidney cortex 813:831 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 3 20 theme organic 530:536 arg1 transporter 544:554 human organic anion transporter 1 524:556 human organic anion transporter 1 (hOAT1) 524:564 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 3 20 theme organic 530:536 arg1 hOAT1 559:563 hOAT1 559:563 hOAT1 559:563 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 1 21 theme clinical 166:173 arg1 effect 183:188 the dose-limiting clinical adverse effect 148:188 the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics 148:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 21 theme clinical 166:173 arg1 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 0 22 theme renal 95:99 arg1 transporter 115:125 renal organic anion transporter 1 95:127 renal organic anion transporter 1 95:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 9 23 dep cidofovir 1352:1360 arg1 m 1378:1378 m 1378:1378 m 1378:1378 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 9 23 dep cidofovir 1352:1360 arg1 K 1376:1376 K 1376:1376 K(m) = 46 and 30 microM, respectively 1376:1412 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 9 24 theme = 1381:1381 arg1 m 1378:1378 m 1378:1378 m 1378:1378 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 9 24 theme = 1381:1381 arg1 K 1376:1376 K 1376:1376 K(m) = 46 and 30 microM, respectively 1376:1412 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 3 25 theme anion 538:542 arg1 transporter 544:554 human organic anion transporter 1 524:556 human organic anion transporter 1 (hOAT1) 524:564 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 3 25 theme anion 538:542 arg1 hOAT1 559:563 hOAT1 559:563 hOAT1 559:563 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 1 26 theme adverse 175:181 arg1 effect 183:188 the dose-limiting clinical adverse effect 148:188 the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics 148:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 26 theme adverse 175:181 arg1 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 0 27 theme antiviral 4:12 arg1 analogs 25:31 The antiviral nucleotide analogs 0:31 The antiviral nucleotide analogs cidofovir and adefovir 0:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 27 theme antiviral 4:12 arg1 adefovir 47:54 adefovir 47:54 adefovir 47:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 27 theme antiviral 4:12 arg1 substrates 66:75 novel substrates 60:75 novel substrates for human and rat renal organic anion transporter 1 60:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 27 theme antiviral 4:12 arg1 cidofovir 33:41 cidofovir 33:41 cidofovir 33:41 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 10 28 from accumulation 1570:1581 arg1 tubules 1627:1633 renal proximal tubules 1612:1633 renal proximal tubules 1612:1633 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 0 29 dep human 81:85 arg1 transporter 115:125 renal organic anion transporter 1 95:127 renal organic anion transporter 1 95:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 7 30 theme = 1054:1054 arg1 microM 1058:1063 K(m) = 4 microM 1049:1063 K(m) = 4 microM 1049:1063 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 7 30 theme = 1054:1054 arg1 p-aminohippurate 1031:1046 p-aminohippurate 1031:1046 p-aminohippurate (K(m) = 4 microM) 1031:1064 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 4 31 theme human 648:652 arg1 kidney 654:659 human kidney 648:659 human kidney 648:659 Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. 10462545 8 32 theme organic 1169:1175 arg1 transporter 1183:1193 rat renal organic anion transporter 1 1159:1195 rat renal organic anion transporter 1 (rROAT1) 1159:1204 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 8 32 theme organic 1169:1175 arg1 rROAT1 1198:1203 rROAT1 1198:1203 rROAT1 1198:1203 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 8 33 theme rat 1159:1161 arg1 transporter 1183:1193 rat renal organic anion transporter 1 1159:1195 rat renal organic anion transporter 1 (rROAT1) 1159:1204 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 8 33 theme rat 1159:1161 arg1 rROAT1 1198:1203 rROAT1 1198:1203 rROAT1 1198:1203 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 6 34 theme Immunoblot 790:799 arg1 analysis 801:808 Immunoblot analysis 790:808 Immunoblot analysis of human kidney cortex 790:831 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 6 35 theme 90-kilodalton 870:882 arg1 protein 898:904 an 80- to 90-kilodalton heterogeneous protein 860:904 an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation 860:941 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 6 35 theme 90-kilodalton 870:882 arg1 hOAT1 851:855 hOAT1 851:855 hOAT1 851:855 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 10 36 theme adefovir 1600:1607 arg1 accumulation 1570:1581 the accumulation 1566:1581 the accumulation of cidofovir and adefovir in renal proximal tubules 1566:1633 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 7 37 theme probenecid-sensitive 994:1013 arg1 uptake 1015:1020 probenecid-sensitive uptake 994:1020 probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate 994:1128 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 8 38 theme other 1282:1286 arg1 antivirals 1311:1320 other nucleoside phosphonate antivirals 1282:1320 other nucleoside phosphonate antivirals 1282:1320 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 1 39 theme cidofovir 193:201 arg1 effect 183:188 the dose-limiting clinical adverse effect 148:188 the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics 148:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 39 theme cidofovir 193:201 arg1 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 0 40 theme nucleotide 14:23 arg1 analogs 25:31 The antiviral nucleotide analogs 0:31 The antiviral nucleotide analogs cidofovir and adefovir 0:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 40 theme nucleotide 14:23 arg1 adefovir 47:54 adefovir 47:54 adefovir 47:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 40 theme nucleotide 14:23 arg1 substrates 66:75 novel substrates 60:75 novel substrates for human and rat renal organic anion transporter 1 60:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 40 theme nucleotide 14:23 arg1 cidofovir 33:41 cidofovir 33:41 cidofovir 33:41 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 9 41 theme hOAT1 1339:1343 arg1 affinity 1327:1334 The affinity 1323:1334 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) 1323:1413 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 9 41 theme hOAT1 1339:1343 arg1 higher 1432:1437 higher 1432:1437 higher 1432:1437 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 0 42 theme anion 109:113 arg1 transporter 115:125 renal organic anion transporter 1 95:127 renal organic anion transporter 1 95:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 6 43 theme heterogeneous 884:896 arg1 protein 898:904 an 80- to 90-kilodalton heterogeneous protein 860:904 an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation 860:941 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 6 43 theme heterogeneous 884:896 arg1 hOAT1 851:855 hOAT1 851:855 hOAT1 851:855 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 10 44 theme cidofovir 1586:1594 arg1 accumulation 1570:1581 the accumulation 1566:1581 the accumulation of cidofovir and adefovir in renal proximal tubules 1566:1633 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 3 45 theme renal 495:499 arg1 library 501:507 a human renal library 487:507 a human renal library 487:507 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 3 46 theme cDNA 458:461 arg1 clone 463:467 A cDNA clone 456:467 A cDNA clone 456:467 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 7 47 dep H 1029:1029 arg1 3 1027:1027 3 1027:1027 3 1027:1027 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 1 48 theme adefovir 207:214 arg1 effect 183:188 the dose-limiting clinical adverse effect 148:188 the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics 148:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 48 theme adefovir 207:214 arg1 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity 130:143 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 8 49 theme saturable 1215:1223 arg1 uptake 1247:1252 saturable, probenecid-sensitive uptake 1215:1252 uptake 1247:1252 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 9 50 dep 5- 1419:1420 arg1 to 1422:1423 to 1422:1423 to 1422:1423 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 8 51 theme phosphonate 1299:1309 arg1 antivirals 1311:1320 other nucleoside phosphonate antivirals 1282:1320 other nucleoside phosphonate antivirals 1282:1320 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 4 52 theme hOAT1 623:627 arg1 transcript 629:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. 10462545 2 53 theme capable 408:414 arg1 transporter 396:406 a renal transporter 388:406 a renal transporter capable of interacting with these therapeutics 388:453 Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. 10462545 5 54 theme reverse 671:677 arg1 reaction 710:717 reverse transcription-polymerase chain reaction 671:717 reverse transcription-polymerase chain reaction 671:717 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 10 55 theme nephrotoxicity 1686:1699 arg1 mechanism 1673:1681 the mechanism 1669:1681 the mechanism of nephrotoxicity associated with these antiviral therapeutics 1669:1744 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 7 56 theme K 1049:1049 arg1 microM 1058:1063 K(m) = 4 microM 1049:1063 K(m) = 4 microM 1049:1063 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 7 56 theme K 1049:1049 arg1 p-aminohippurate 1031:1046 p-aminohippurate 1031:1046 p-aminohippurate (K(m) = 4 microM) 1031:1064 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 2 57 theme nephrotoxicity 347:360 arg1 etiology 300:307 the etiology 296:307 the etiology of cidofovir- and adefovir-associated nephrotoxicity 296:360 Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. 10462545 4 58 theme specific 596:603 arg1 transcript 629:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. 10462545 5 59 theme transcription-polymerase 679:702 arg1 reaction 710:717 reverse transcription-polymerase chain reaction 671:717 reverse transcription-polymerase chain reaction 671:717 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 2 60 theme adefovir-associated 327:345 arg1 nephrotoxicity 347:360 cidofovir- and adefovir-associated nephrotoxicity 312:360 cidofovir- and adefovir-associated nephrotoxicity 312:360 Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. 10462545 7 61 theme Xenopus 944:950 arg1 oocytes 959:965 Xenopus laevis oocytes 944:965 Xenopus laevis oocytes expressing hOAT1 944:982 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 6 62 dep 90-kilodalton 870:882 arg1 to 867:868 to 867:868 to 867:868 Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. 10462545 5 63 theme chain 704:708 arg1 reaction 710:717 reverse transcription-polymerase chain reaction 671:717 reverse transcription-polymerase chain reaction 671:717 However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. 10462545 10 64 dep contribute 1552:1561 arg1 play 1646:1649 play 1646:1649 play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics 1646:1744 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 0 65 dep analogs 25:31 arg1 analogs 25:31 The antiviral nucleotide analogs 0:31 The antiviral nucleotide analogs cidofovir and adefovir 0:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 65 dep analogs 25:31 arg1 adefovir 47:54 adefovir 47:54 adefovir 47:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 65 dep analogs 25:31 arg1 cidofovir 33:41 cidofovir 33:41 cidofovir 33:41 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 7 66 theme p-aminohippurate 1031:1046 arg1 uptake 1015:1020 probenecid-sensitive uptake 994:1020 probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate 994:1128 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 8 67 theme nucleoside 1288:1297 arg1 antivirals 1311:1320 other nucleoside phosphonate antivirals 1282:1320 other nucleoside phosphonate antivirals 1282:1320 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 7 68 theme laevis 952:957 arg1 oocytes 959:965 Xenopus laevis oocytes 944:965 Xenopus laevis oocytes expressing hOAT1 944:982 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 4 69 theme 2.5-kilobase 605:616 arg1 transcript 629:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. 10462545 7 70 theme m 1051:1051 arg1 microM 1058:1063 K(m) = 4 microM 1049:1063 K(m) = 4 microM 1049:1063 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 7 70 theme m 1051:1051 arg1 p-aminohippurate 1031:1046 p-aminohippurate 1031:1046 p-aminohippurate (K(m) = 4 microM) 1031:1064 Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. 10462545 9 71 dep = 1381:1381 arg1 microM 1393:1398 46 and 30 microM 1383:1398 46 and 30 microM 1383:1398 The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). 10462545 2 72 theme renal 260:264 arg1 uptake 266:271 renal uptake 260:271 renal uptake 260:271 Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. 10462545 2 73 theme cidofovir- 312:321 arg1 nephrotoxicity 347:360 cidofovir- and adefovir-associated nephrotoxicity 312:360 cidofovir- and adefovir-associated nephrotoxicity 312:360 Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. 10462545 1 74 theme potent 221:226 arg1 therapeutics 238:249 two potent antiviral therapeutics 217:249 two potent antiviral therapeutics 217:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 74 theme potent 221:226 arg1 cidofovir 193:201 cidofovir 193:201 cidofovir 193:201 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 10 75 theme renal 1612:1616 arg1 tubules 1627:1633 renal proximal tubules 1612:1633 renal proximal tubules 1612:1633 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 10 76 theme antiviral 1723:1731 arg1 therapeutics 1733:1744 these antiviral therapeutics 1717:1744 these antiviral therapeutics 1717:1744 These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. 10462545 8 77 theme cidofovir 1257:1265 arg1 uptake 1247:1252 saturable, probenecid-sensitive uptake 1215:1252 uptake 1247:1252 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 8 78 theme antivirals 1311:1320 arg1 uptake 1247:1252 saturable, probenecid-sensitive uptake 1215:1252 uptake 1247:1252 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 2 79 theme renal 390:394 arg1 transporter 396:406 a renal transporter 388:406 a renal transporter capable of interacting with these therapeutics 388:453 Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. 10462545 1 80 theme antiviral 228:236 arg1 therapeutics 238:249 two potent antiviral therapeutics 217:249 two potent antiviral therapeutics 217:249 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 1 80 theme antiviral 228:236 arg1 cidofovir 193:201 cidofovir 193:201 cidofovir 193:201 Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. 10462545 0 81 theme novel 60:64 arg1 analogs 25:31 The antiviral nucleotide analogs 0:31 The antiviral nucleotide analogs cidofovir and adefovir 0:54 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 0 81 theme novel 60:64 arg1 substrates 66:75 novel substrates 60:75 novel substrates for human and rat renal organic anion transporter 1 60:127 The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. 10462545 8 82 theme adefovir 1268:1275 arg1 uptake 1247:1252 saturable, probenecid-sensitive uptake 1215:1252 uptake 1247:1252 Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid-sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. 10462545 3 83 theme human 524:528 arg1 transporter 544:554 human organic anion transporter 1 524:556 human organic anion transporter 1 (hOAT1) 524:564 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 3 83 theme human 524:528 arg1 hOAT1 559:563 hOAT1 559:563 hOAT1 559:563 A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). 10462545 4 84 theme pair 618:621 arg1 transcript 629:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 a specific 2.5-kilobase pair hOAT1 transcript 594:638 Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. 7681597 4 0 located found 666:670 arg1 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 0 located found 666:670 arg1 three 715:719 three 715:719 three 715:719 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 0 located found 666:670 arg2 antigenicity 578:589 The Tn antigenicity 571:589 The Tn antigenicity 571:589 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 0 located found 666:670 arg1 glycopeptides 691:703 the glycopeptides 687:703 the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr 687:791 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 0 located found 666:670 arg1 four 736:739 four 736:739 four 736:739 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 2 1 gly glycosylated 382:393 arg1 Ser-15 366:371 Ser-15 366:371 Ser-15 366:371 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 1 gly glycosylated 382:393 arg1 Ser-14 355:360 Ser-14 355:360 Ser-14 355:360 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 1 gly glycosylated 382:393 arg1 residues 345:352 two serine residues 334:352 two serine residues (Ser-14 and Ser-15) 334:372 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 2 theme serine 338:343 arg1 Ser-15 366:371 Ser-15 366:371 Ser-15 366:371 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 2 theme serine 338:343 arg1 Ser-14 355:360 Ser-14 355:360 Ser-14 355:360 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 2 theme serine 338:343 arg1 residues 345:352 two serine residues 334:352 two serine residues (Ser-14 and Ser-15) 334:372 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 1 3 theme chromatographies 219:234 arg1 combination 183:193 a combination 181:193 a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6 181:278 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 7681597 4 4 theme GalNAc-Ser/Thr 857:870 arg1 residues 872:879 two nonconsecutive GalNAc-Ser/Thr residues 838:879 two nonconsecutive GalNAc-Ser/Thr residues 838:879 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 8 5 theme GPA-6 1142:1146 arg1 immunoreactivity 1113:1128 the immunoreactivity 1109:1128 the immunoreactivity of GPA-5 or GPA-6 1109:1146 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 5 6 theme GPA-1 943:947 arg1 immunoreactivities 921:938 The immunoreactivities 917:938 The immunoreactivities 917:938 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 6 theme GPA-1 943:947 arg1 % 1036:1036 63% 1034:1036 63% (calcd 1034:1043 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 4 7 theme Tn 900:901 arg1 antigenicity 903:914 practically no Tn antigenicity 885:914 practically no Tn antigenicity 885:914 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 5 8 theme GPA-3 953:957 arg1 immunoreactivities 921:938 The immunoreactivities 917:938 The immunoreactivities 917:938 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 8 theme GPA-3 953:957 arg1 % 1036:1036 63% 1034:1036 63% (calcd 1034:1043 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 10 9 theme ovine 1494:1498 arg1 mucin 1513:1517 ovine submaxillary mucin 1494:1517 ovine submaxillary mucin 1494:1517 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 3 10 gly glycopeptides 491:503 arg2 glycopeptides 491:503 The glycopeptides 487:503 The glycopeptides thus obtained 487:517 The glycopeptides thus obtained were treated with sialidase and beta-galactosidase. 7681597 10 11 theme Tn 1441:1442 arg1 antigenicity 1444:1455 Tn antigenicity 1441:1455 Tn antigenicity in the light of previous results for ovine submaxillary mucin 1441:1517 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 4 12 gly glycopeptides 691:703 arg2 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 12 gly glycopeptides 691:703 arg2 three 715:719 three 715:719 three 715:719 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 12 gly glycopeptides 691:703 arg2 four 736:739 four 736:739 four 736:739 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 12 gly glycopeptides 691:703 arg2 glycopeptides 691:703 the glycopeptides 687:703 the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr 687:791 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 10 13 gly glycosylated 1390:1401 arg1 residues 1411:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 10 14 theme submaxillary 1500:1511 arg1 mucin 1513:1517 ovine submaxillary mucin 1494:1517 ovine submaxillary mucin 1494:1517 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 4 15 theme Tn 575:576 arg1 antigenicity 578:589 The Tn antigenicity 571:589 The Tn antigenicity 571:589 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 8 16 contain containing 1149:1158 arg1 immunoreactivity 1113:1128 the immunoreactivity 1109:1128 the immunoreactivity of GPA-5 or GPA-6 1109:1146 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 8 16 contain containing 1149:1158 arg1 % 1088:1088 50%) 1086:1089 50%) 1086:1089 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 8 16 contain containing 1149:1158 arg2 I 1168:1168 cluster I 1160:1168 cluster I (the average being taken as the basis) 1160:1207 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 4 17 theme GalNAc-Ser/Thr 778:791 arg1 three 715:719 three 715:719 three 715:719 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 17 theme GalNAc-Ser/Thr 778:791 arg1 I 730:730 cluster I 722:730 cluster I 722:730 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 17 theme GalNAc-Ser/Thr 778:791 arg1 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 8 18 dep I 1168:1168 arg1 average 1175:1181 the average 1171:1181 the average being taken as the basis 1171:1206 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 4 19 theme anti-Tn 634:640 arg1 MLS 652:654 MLS 128 652:658 MLS 128 652:658 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 19 theme anti-Tn 634:640 arg1 antibody 642:649 a monoclonal anti-Tn antibody 621:649 a monoclonal anti-Tn antibody (MLS 128) 621:659 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 2 20 dep residues 345:352 arg1 Ser-15 366:371 Ser-15 366:371 Ser-15 366:371 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 20 dep residues 345:352 arg1 Ser-14 355:360 Ser-14 355:360 Ser-14 355:360 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 20 dep residues 345:352 arg1 residues 345:352 two serine residues 334:352 two serine residues (Ser-14 and Ser-15) 334:372 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 4 21 gly glycopeptide 806:817 arg2 GPA-2 820:824 GPA-2 820:824 GPA-2 820:824 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 21 gly glycopeptide 806:817 arg2 glycopeptide 806:817 the glycopeptide 802:817 the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues 802:879 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 10 22 theme consecutive 1378:1388 arg1 residues 1411:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 9 23 theme same 1331:1334 arg1 extent 1336:1341 the same extent 1327:1341 the same extent 1327:1341 These results indicate that clusters I and II react with the antibody to the same extent. 7681597 1 24 dep Pasteurella 121:131 arg1 haemolytica 133:143 Pasteurella haemolytica 121:143 Pasteurella haemolytica 121:143 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 7681597 4 25 contain containing 827:836 arg1 glycopeptide 806:817 the glycopeptide 802:817 the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues 802:879 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 25 contain containing 827:836 arg2 residues 872:879 two nonconsecutive GalNAc-Ser/Thr residues 838:879 two nonconsecutive GalNAc-Ser/Thr residues 838:879 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 25 contain containing 827:836 arg1 GPA-2 820:824 GPA-2 820:824 GPA-2 820:824 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 0 26 theme Epitopic 0:7 arg1 structure 9:17 Epitopic structure 0:17 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 0:72 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 7681597 4 27 contain had 881:883 arg1 glycopeptide 806:817 the glycopeptide 802:817 the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues 802:879 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 27 contain had 881:883 arg1 GPA-2 820:824 GPA-2 820:824 GPA-2 820:824 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 27 contain had 881:883 arg2 antigenicity 903:914 practically no Tn antigenicity 885:914 practically no Tn antigenicity 885:914 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 5 28 dep clusters 976:983 arg1 clusters 976:983 both clusters I and II 971:992 both clusters I and II 971:992 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 28 dep clusters 976:983 arg1 II 991:992 II 991:992 II 991:992 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 28 dep clusters 976:983 arg1 I 985:985 I 985:985 I 985:985 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 0 29 theme Tn 22:23 arg1 A 37:37 Tn glycophorin A 22:37 Tn glycophorin A 22:37 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 7681597 1 30 theme GPA-1 265:269 arg1 glycopeptides 251:263 the glycopeptides 247:263 the glycopeptides GPA-1 to GPA-6 247:278 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 7681597 4 31 theme consecutive 754:764 arg1 residues 766:773 consecutive residues 754:773 consecutive residues 754:773 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 2 32 theme Sequence 281:288 arg1 analysis 290:297 Sequence analysis 281:297 Sequence analysis of the glycopeptides 281:318 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 33 dep structure 476:484 arg1 disagreement 445:456 disagreement 445:456 disagreement 445:456 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 4 34 dep antigenicity 578:589 arg1 assayed 595:601 assayed 595:601 assayed by the binding to a monoclonal anti-Tn antibody (MLS 128) 595:659 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 1 35 theme Glycophorin 74:84 arg1 A 86:86 Glycophorin A 74:86 Glycophorin A 74:86 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 7681597 6 36 dep % 1054:1054 arg1 calcd 1057:1061 calcd 1057:1061 calcd 1057:1061 67%), 81% (calcd. 7681597 0 37 theme A 37:37 arg1 structure 9:17 Epitopic structure 0:17 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 0:72 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 7681597 7 38 dep % 1076:1076 arg1 calcd 1079:1083 calcd 1079:1083 calcd 1079:1083 86%), and 50% (calcd. 7681597 9 39 dep clusters 1282:1289 arg1 I 1291:1291 I 1291:1291 I 1291:1291 These results indicate that clusters I and II react with the antibody to the same extent. 7681597 9 39 dep clusters 1282:1289 arg1 clusters 1282:1289 clusters I and II 1282:1298 clusters I and II 1282:1298 These results indicate that clusters I and II react with the antibody to the same extent. 7681597 9 39 dep clusters 1282:1289 arg1 II 1297:1298 II 1297:1298 II 1297:1298 These results indicate that clusters I and II react with the antibody to the same extent. 7681597 0 40 theme glycophorin 25:35 arg1 A 37:37 Tn glycophorin A 22:37 Tn glycophorin A 22:37 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 7681597 5 41 contain containing 1006:1015 arg1 immunoreactivities 921:938 The immunoreactivities 917:938 The immunoreactivities 917:938 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 41 contain containing 1006:1015 arg2 cluster 1017:1023 cluster II 1017:1026 cluster II 1017:1026 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 41 contain containing 1006:1015 arg1 % 1036:1036 63% 1034:1036 63% (calcd 1034:1043 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 10 42 theme Ser/Thr 1403:1409 arg1 residues 1411:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 5 43 dep % 1036:1036 arg1 calcd 1039:1043 calcd 1039:1043 calcd 1039:1043 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 8 44 theme cluster 1160:1166 arg1 I 1168:1168 cluster I 1160:1168 cluster I (the average being taken as the basis) 1160:1207 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 8 45 theme GalNAc 1238:1243 arg1 residue 1245:1251 GalNAc residue 1238:1251 GalNAc residue 1238:1251 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 2 46 gly glycosylated 420:431 arg1 Ser-19 407:412 Ser-19 407:412 Ser-19 407:412 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 46 gly glycosylated 420:431 arg1 Ser-15 366:371 Ser-15 366:371 Ser-15 366:371 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 46 gly glycosylated 420:431 arg1 Thr-17 396:401 Thr-17 396:401 Thr-17 396:401 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 46 gly glycosylated 420:431 arg1 residues 345:352 two serine residues 334:352 two serine residues (Ser-14 and Ser-15) 334:372 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 46 gly glycosylated 420:431 arg1 Ser-14 355:360 Ser-14 355:360 Ser-14 355:360 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 1 47 gly glycopeptides 251:263 arg2 glycopeptides 251:263 the glycopeptides 247:263 the glycopeptides GPA-1 to GPA-6 247:278 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 7681597 4 48 dep three 715:719 arg1 residues 766:773 consecutive residues 754:773 consecutive residues 754:773 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 10 49 theme glycosylated 1390:1401 arg1 residues 1411:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 three consecutive glycosylated Ser/Thr residues 1372:1418 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 2 50 dep glycosylated 382:393 arg1 glycosylated 420:431 glycosylated 420:431 being glycosylated instead 414:439 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 10 51 theme results 1482:1488 arg1 light 1464:1468 the light 1460:1468 the light of previous results for ovine submaxillary mucin 1460:1517 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 4 52 theme nonconsecutive 842:855 arg1 residues 872:879 two nonconsecutive GalNAc-Ser/Thr residues 838:879 two nonconsecutive GalNAc-Ser/Thr residues 838:879 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 2 53 theme glycopeptides 306:318 arg1 analysis 290:297 Sequence analysis 281:297 Sequence analysis of the glycopeptides 281:318 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 2 54 gly glycopeptides 306:318 arg2 glycopeptides 306:318 the glycopeptides 302:318 the glycopeptides 302:318 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 4 55 theme cluster 722:728 arg1 three 715:719 three 715:719 three 715:719 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 55 theme cluster 722:728 arg1 I 730:730 cluster I 722:730 cluster I 722:730 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 55 theme cluster 722:728 arg1 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 GalNAc-Ser/Thr 778:791 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 8 56 theme immunoreactivity 1113:1128 arg1 immunoreactivity 1113:1128 the immunoreactivity 1109:1128 the immunoreactivity of GPA-5 or GPA-6 1109:1146 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 8 56 theme immunoreactivity 1113:1128 arg1 % 1088:1088 50%) 1086:1089 50%) 1086:1089 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 10 57 theme previous 1473:1480 arg1 results 1482:1488 previous results 1473:1488 previous results for ovine submaxillary mucin 1473:1517 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 2 58 theme accepted 467:474 arg1 structure 476:484 the accepted structure 463:484 the accepted structure 463:484 Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. 7681597 5 59 theme GPA-4 999:1003 arg1 immunoreactivities 921:938 The immunoreactivities 917:938 The immunoreactivities 917:938 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 59 theme GPA-4 999:1003 arg1 % 1036:1036 63% 1034:1036 63% (calcd 1034:1043 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 10 60 from antigenicity 1444:1455 arg1 light 1464:1468 the light 1460:1468 the light of previous results for ovine submaxillary mucin 1460:1517 The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin. 7681597 4 61 theme monoclonal 623:632 arg1 MLS 652:654 MLS 128 652:658 MLS 128 652:658 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 4 61 theme monoclonal 623:632 arg1 antibody 642:649 a monoclonal anti-Tn antibody 621:649 a monoclonal anti-Tn antibody (MLS 128) 621:659 The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. 7681597 1 62 theme high-pressure 198:210 arg1 chromatographies 219:234 high-pressure column chromatographies 198:234 high-pressure column chromatographies 198:234 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 7681597 0 63 theme anti-Tn 46:52 arg1 MLS 64:66 MLS 128 64:70 MLS 128 64:70 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 7681597 0 63 theme anti-Tn 46:52 arg1 antibody 54:61 an anti-Tn antibody 43:61 an anti-Tn antibody (MLS 128) 43:71 Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 7681597 8 64 theme GPA-5 1133:1137 arg1 immunoreactivity 1113:1128 the immunoreactivity 1109:1128 the immunoreactivity of GPA-5 or GPA-6 1109:1146 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. 7681597 5 65 contain containing 960:969 arg2 II 991:992 II 991:992 II 991:992 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 65 contain containing 960:969 arg2 clusters 976:983 both clusters I and II 971:992 both clusters I and II 971:992 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 65 contain containing 960:969 arg2 I 985:985 I 985:985 I 985:985 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 5 65 contain containing 960:969 arg1 GPA-1 943:947 GPA-1 943:947 GPA-1 943:947 The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 7681597 1 66 theme column 212:217 arg1 chromatographies 219:234 high-pressure column chromatographies 198:234 high-pressure column chromatographies 198:234 Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. 21572394 7 0 theme biosynthesis 1162:1173 arg1 regulation 1139:1148 the regulation 1135:1148 the regulation of dolichol biosynthesis 1135:1173 Identification of NgBR as a component of the cis-IPTase machinery yields insights into the regulation of dolichol biosynthesis. 21572394 5 1 from reduction 883:891 arg1 N-glycosylation 904:918 protein N-glycosylation 896:918 protein N-glycosylation 896:918 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 1 2 from carrier 158:164 arg1 reactions 197:205 protein glycosylation reactions 175:205 protein glycosylation reactions 175:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 4 3 theme machinery 709:717 arg1 component 673:681 an essential component 660:681 an essential component of the Dol-P biosynthetic machinery 660:717 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 4 3 theme machinery 709:717 arg1 receptor 641:648 Nogo-B receptor 634:648 Nogo-B receptor (NgBR) 634:655 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 6 4 theme Dol-P 1030:1034 arg1 production 1036:1045 Dol-P production 1030:1045 Dol-P production 1030:1045 NgBR interacts with the previously identified cis-IPTase hCIT, enhances hCIT protein stability, and promotes Dol-P production. 21572394 5 5 link dolichol-linked 838:852 arg1 oligosaccharides 854:869 dolichol-linked oligosaccharides 838:869 dolichol-linked oligosaccharides 838:869 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 1 6 dep carrier 158:164 arg1 lipid 166:170 lipid 166:170 an obligate glycosyl carrier lipid in protein glycosylation reactions 137:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 5 7 from levels 828:833 arg1 N-glycosylation 904:918 protein N-glycosylation 896:918 protein N-glycosylation 896:918 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 7 8 theme dolichol 1153:1160 arg1 biosynthesis 1162:1173 dolichol biosynthesis 1153:1173 dolichol biosynthesis 1153:1173 Identification of NgBR as a component of the cis-IPTase machinery yields insights into the regulation of dolichol biosynthesis. 21572394 5 9 theme broad 877:881 arg1 reduction 883:891 a broad reduction 875:891 a broad reduction in protein N-glycosylation 875:918 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 5 10 theme robust 746:751 arg1 deficit 753:759 a robust deficit 744:759 a robust deficit in cis-IPTase activity and Dol-P production 744:803 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 3 11 theme mammalian 500:508 arg1 enzyme 510:515 the mammalian enzyme 496:515 the mammalian enzyme 496:515 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 5 12 theme oligosaccharides 854:869 arg1 reduction 883:891 a broad reduction 875:891 a broad reduction in protein N-glycosylation 875:918 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 5 12 theme oligosaccharides 854:869 arg1 levels 828:833 diminished levels 817:833 diminished levels of dolichol-linked oligosaccharides 817:869 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 3 13 theme responsible 542:552 arg1 machinery 532:540 the molecular machinery 518:540 the molecular machinery responsible for regulating this activity 518:581 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 3 14 theme molecular 451:459 arg1 cloning 461:467 the molecular cloning 447:467 the molecular cloning of the human cDNA encoding the mammalian enzyme 447:515 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 5 15 theme Dol-P 788:792 arg1 production 794:803 Dol-P production 788:803 Dol-P production 788:803 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 3 16 theme cis-IPTase 410:419 arg1 activity 421:428 cis-IPTase activity 410:428 cis-IPTase activity 40 years ago 410:441 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 1 17 theme protein 175:181 arg1 reactions 197:205 protein glycosylation reactions 175:205 protein glycosylation reactions 175:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 1 18 theme Dolichol 93:100 arg1 monophosphate 102:114 Dolichol monophosphate 93:114 Dolichol monophosphate (Dol-P) 93:122 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 1 18 theme Dolichol 93:100 arg1 carrier 158:164 an obligate glycosyl carrier 137:164 an obligate glycosyl carrier lipid in protein glycosylation reactions 137:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 1 18 theme Dolichol 93:100 arg1 Dol-P 117:121 Dol-P 117:121 Dol-P 117:121 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 1 19 theme glycosylation 183:195 arg1 reactions 197:205 protein glycosylation reactions 175:205 protein glycosylation reactions 175:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 0 20 theme Nogo-B 0:5 arg1 receptor 7:14 Nogo-B receptor 0:14 Nogo-B receptor 0:14 Nogo-B receptor is necessary for cellular dolichol biosynthesis and protein N-glycosylation. 21572394 5 21 theme protein 896:902 arg1 N-glycosylation 904:918 protein N-glycosylation 896:918 protein N-glycosylation 896:918 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 5 22 theme diminished 817:826 arg1 levels 828:833 diminished levels 817:833 diminished levels of dolichol-linked oligosaccharides 817:869 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 7 23 theme machinery 1104:1112 arg1 component 1076:1084 a component 1074:1084 a component of the cis-IPTase machinery 1074:1112 Identification of NgBR as a component of the cis-IPTase machinery yields insights into the regulation of dolichol biosynthesis. 21572394 3 24 theme human 476:480 arg1 cDNA 482:485 the human cDNA 472:485 the human cDNA encoding the mammalian enzyme 472:515 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 6 25 theme protein 998:1004 arg1 stability 1006:1014 hCIT protein stability 993:1014 hCIT protein stability 993:1014 NgBR interacts with the previously identified cis-IPTase hCIT, enhances hCIT protein stability, and promotes Dol-P production. 21572394 2 26 theme farnesyl 299:306 arg1 diphosphate 308:318 farnesyl diphosphate 299:318 farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity 299:380 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 3 27 theme cDNA 482:485 arg1 cloning 461:467 the molecular cloning 447:467 the molecular cloning of the human cDNA encoding the mammalian enzyme 447:515 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 3 27 theme cDNA 482:485 arg1 recognition 395:405 the recognition 391:405 the recognition of cis-IPTase activity 40 years ago 391:441 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 0 28 theme cellular 33:40 arg1 biosynthesis 51:62 cellular dolichol biosynthesis 33:62 cellular dolichol biosynthesis 33:62 Nogo-B receptor is necessary for cellular dolichol biosynthesis and protein N-glycosylation. 21572394 6 29 theme hCIT 993:996 arg1 stability 1006:1014 hCIT protein stability 993:1014 hCIT protein stability 993:1014 NgBR interacts with the previously identified cis-IPTase hCIT, enhances hCIT protein stability, and promotes Dol-P production. 21572394 5 30 theme NgBR 728:731 arg1 Loss 720:723 Loss 720:723 Loss of NgBR 720:731 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 5 31 from deficit 753:759 arg1 activity 775:782 cis-IPTase activity 764:782 cis-IPTase activity 764:782 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 5 31 from deficit 753:759 arg1 production 794:803 Dol-P production 788:803 Dol-P production 788:803 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 4 32 theme biosynthetic 696:707 arg1 machinery 709:717 the Dol-P biosynthetic machinery 686:717 the Dol-P biosynthetic machinery 686:717 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 7 33 theme cis-IPTase 1093:1102 arg1 machinery 1104:1112 the cis-IPTase machinery 1089:1112 the cis-IPTase machinery 1089:1112 Identification of NgBR as a component of the cis-IPTase machinery yields insights into the regulation of dolichol biosynthesis. 21572394 2 34 theme successive 236:245 arg1 condensation 247:258 the successive condensation 232:258 the successive condensation of isopentenyl diphosphate (IPP) 232:291 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 3 35 theme activity 421:428 arg1 cloning 461:467 the molecular cloning 447:467 the molecular cloning of the human cDNA encoding the mammalian enzyme 447:515 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 3 35 theme activity 421:428 arg1 recognition 395:405 the recognition 391:405 the recognition of cis-IPTase activity 40 years ago 391:441 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 7 36 theme NgBR 1066:1069 arg1 Identification 1048:1061 Identification 1048:1061 Identification of NgBR as a component of the cis-IPTase machinery 1048:1112 Identification of NgBR as a component of the cis-IPTase machinery yields insights into the regulation of dolichol biosynthesis. 21572394 2 37 theme cis-IPTase 361:370 arg1 activity 373:380 a cis-isoprenyltransferase (cis-IPTase) activity 333:380 a cis-isoprenyltransferase (cis-IPTase) activity 333:380 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 0 38 theme dolichol 42:49 arg1 biosynthesis 51:62 cellular dolichol biosynthesis 33:62 cellular dolichol biosynthesis 33:62 Nogo-B receptor is necessary for cellular dolichol biosynthesis and protein N-glycosylation. 21572394 6 39 theme cis-IPTase 967:976 arg1 hCIT 978:981 the previously identified cis-IPTase hCIT 941:981 the previously identified cis-IPTase hCIT 941:981 NgBR interacts with the previously identified cis-IPTase hCIT, enhances hCIT protein stability, and promotes Dol-P production. 21572394 1 40 theme obligate 140:147 arg1 monophosphate 102:114 Dolichol monophosphate 93:114 Dolichol monophosphate (Dol-P) 93:122 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 1 40 theme obligate 140:147 arg1 carrier 158:164 an obligate glycosyl carrier 137:164 an obligate glycosyl carrier lipid in protein glycosylation reactions 137:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 2 41 theme cis-isoprenyltransferase 335:358 arg1 activity 373:380 a cis-isoprenyltransferase (cis-IPTase) activity 333:380 a cis-isoprenyltransferase (cis-IPTase) activity 333:380 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 4 42 theme Nogo-B 634:639 arg1 component 673:681 an essential component 660:681 an essential component of the Dol-P biosynthetic machinery 660:717 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 4 42 theme Nogo-B 634:639 arg1 NgBR 651:654 NgBR 651:654 NgBR 651:654 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 4 42 theme Nogo-B 634:639 arg1 receptor 641:648 Nogo-B receptor 634:648 Nogo-B receptor (NgBR) 634:655 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 2 43 theme diphosphate 275:285 arg1 condensation 247:258 the successive condensation 232:258 the successive condensation of isopentenyl diphosphate (IPP) 232:291 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 4 44 theme Dol-P 690:694 arg1 machinery 709:717 the Dol-P biosynthetic machinery 686:717 the Dol-P biosynthetic machinery 686:717 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 0 45 theme protein 68:74 arg1 N-glycosylation 76:90 protein N-glycosylation 68:90 protein N-glycosylation 68:90 Nogo-B receptor is necessary for cellular dolichol biosynthesis and protein N-glycosylation. 21572394 2 46 theme isopentenyl 263:273 arg1 IPP 288:290 IPP 288:290 IPP 288:290 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 2 46 theme isopentenyl 263:273 arg1 diphosphate 275:285 isopentenyl diphosphate 263:285 isopentenyl diphosphate (IPP) 263:291 Dol-P is synthesized by the successive condensation of isopentenyl diphosphate (IPP), with farnesyl diphosphate catalysed by a cis-isoprenyltransferase (cis-IPTase) activity. 21572394 5 47 theme dolichol-linked 838:852 arg1 oligosaccharides 854:869 dolichol-linked oligosaccharides 838:869 dolichol-linked oligosaccharides 838:869 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 3 48 theme molecular 522:530 arg1 machinery 532:540 the molecular machinery 518:540 the molecular machinery responsible for regulating this activity 518:581 Despite the recognition of cis-IPTase activity 40 years ago and the molecular cloning of the human cDNA encoding the mammalian enzyme, the molecular machinery responsible for regulating this activity remains incompletely understood. 21572394 4 49 theme essential 663:671 arg1 component 673:681 an essential component 660:681 an essential component of the Dol-P biosynthetic machinery 660:717 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 4 49 theme essential 663:671 arg1 receptor 641:648 Nogo-B receptor 634:648 Nogo-B receptor (NgBR) 634:655 Here, we identify Nogo-B receptor (NgBR) as an essential component of the Dol-P biosynthetic machinery. 21572394 5 50 theme cis-IPTase 764:773 arg1 activity 775:782 cis-IPTase activity 764:782 cis-IPTase activity 764:782 Loss of NgBR results in a robust deficit in cis-IPTase activity and Dol-P production, leading to diminished levels of dolichol-linked oligosaccharides and a broad reduction in protein N-glycosylation. 21572394 6 51 theme identified 956:965 arg1 hCIT 978:981 the previously identified cis-IPTase hCIT 941:981 the previously identified cis-IPTase hCIT 941:981 NgBR interacts with the previously identified cis-IPTase hCIT, enhances hCIT protein stability, and promotes Dol-P production. 21572394 1 52 theme glycosyl 149:156 arg1 monophosphate 102:114 Dolichol monophosphate 93:114 Dolichol monophosphate (Dol-P) 93:122 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 21572394 1 52 theme glycosyl 149:156 arg1 carrier 158:164 an obligate glycosyl carrier 137:164 an obligate glycosyl carrier lipid in protein glycosylation reactions 137:205 Dolichol monophosphate (Dol-P) functions as an obligate glycosyl carrier lipid in protein glycosylation reactions. 18596036 2 0 theme -CCP 297:300 arg1 array 312:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array 260:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end 260:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 6 1 dep contributed 1007:1017 arg1 module 980:985 module 980:985 module 980:985 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 8 2 dep sites 1475:1479 arg1 stabilized 1583:1592 stabilized 1583:1592 stabilized by the Ca(2+) ion 1583:1610 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 2 dep sites 1475:1479 arg1 contributed 1481:1491 contributed 1481:1491 contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites 1481:1577 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 4 3 theme CUB 715:717 arg1 module 722:727 the CUB(1) module 711:727 the CUB(1) module of one monomer 711:742 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 8 4 theme Ca 1601:1602 arg1 ion 1608:1610 the Ca(2+) ion 1597:1610 the Ca(2+) ion 1597:1610 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 2 5 theme CUB 273:275 arg1 array 312:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array 260:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end 260:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 1 6 theme homologous 171:180 arg1 proteases 182:190 homologous proteases 171:190 homologous proteases arising from alternative splicing of the MASP1/3 gene 171:244 MASP-1 and MASP-3 are homologous proteases arising from alternative splicing of the MASP1/3 gene. 18596036 1 6 theme homologous 171:180 arg1 MASP-1 149:154 MASP-1 149:154 MASP-1 149:154 MASP-1 and MASP-3 are homologous proteases arising from alternative splicing of the MASP1/3 gene. 18596036 1 6 theme homologous 171:180 arg1 MASP-3 160:165 MASP-3 160:165 MASP-3 160:165 MASP-1 and MASP-3 are homologous proteases arising from alternative splicing of the MASP1/3 gene. 18596036 8 7 theme 2+ 1604:1605 arg1 ion 1608:1610 the Ca(2+) ion 1597:1610 the Ca(2+) ion 1597:1610 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 3 8 theme responsible 459:469 arg1 MASP-1/3 449:456 human MASP-1/3 443:456 human MASP-1/3 443:456 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 4 9 theme factor 769:774 arg1 module 782:787 the epidermal growth factor (EGF) module 748:787 the epidermal growth factor (EGF) module of its counterpart 748:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 3 10 theme 2.3A 604:607 arg1 resolution 590:599 a resolution 588:599 a resolution of 2.3A(.) 588:610 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 0 11 theme mannan-binding 113:126 arg1 lectin 128:133 mannan-binding lectin 113:133 mannan-binding lectin 113:133 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 5 12 theme intra- 873:878 arg1 interfaces 909:918 the intra- and inter-monomer CUB(1)-EGF interfaces 869:918 the intra- and inter-monomer CUB(1)-EGF interfaces 869:918 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 3 13 theme MASP-1/3 449:456 arg1 domain 433:438 the CUB(1)-EGF-CUB(2) domain 411:438 the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, 411:572 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 13 theme MASP-1/3 449:456 arg1 MASP-1/3 449:456 human MASP-1/3 443:456 human MASP-1/3 443:456 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 1 14 theme alternative 205:215 arg1 splicing 217:224 alternative splicing 205:224 alternative splicing of the MASP1/3 gene 205:244 MASP-1 and MASP-3 are homologous proteases arising from alternative splicing of the MASP1/3 gene. 18596036 9 15 theme MBL-MASP-1/3 1666:1677 arg1 interaction 1679:1689 the MBL-MASP-1/3 interaction 1662:1689 the MBL-MASP-1/3 interaction 1662:1689 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 6 16 theme Asp 1031:1033 arg1 Glu 1095:1097 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 5 17 theme inter-monomer 884:896 arg1 -EGF 904:907 inter-monomer CUB(1)-EGF 884:907 inter-monomer CUB(1)-EGF 884:907 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 5 17 theme inter-monomer 884:896 arg1 1 902:902 1 902:902 1 902:902 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 3 18 theme MASP-1 490:495 arg1 interaction 475:485 interaction 475:485 interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins 475:571 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 5 19 theme EGF 846:848 arg1 modules 850:856 both EGF modules 841:856 both EGF modules 841:856 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 7 20 theme human 1314:1318 arg1 MASP-3 1320:1325 human MASP-3 1314:1325 human MASP-3 1314:1325 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 4 21 theme epidermal 752:760 arg1 factor 769:774 epidermal growth factor 752:774 the epidermal growth factor (EGF) module of its counterpart 748:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 4 21 theme epidermal 752:760 arg1 EGF 777:779 EGF 777:779 EGF 777:779 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 10 22 theme complex 1876:1882 arg1 model 1856:1860 a schematic model 1844:1860 a schematic model of a MBL.MASP complex 1844:1882 Based on these and other data, a schematic model of a MBL.MASP complex is proposed. 18596036 9 23 theme interaction 1679:1689 arg1 model 1653:1657 a model 1651:1657 a model 1651:1657 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 6 24 dep CUB 973:975 arg1 contributed 1007:1017 contributed 1007:1017 module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively 980:1193 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 0 25 theme MASP-1/3 55:62 arg1 domain 39:44 the CUB1-EGF-CUB2 domain 21:44 the CUB1-EGF-CUB2 domain of human MASP-1/3 21:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 0 25 theme MASP-1/3 55:62 arg1 MASP-1/3 55:62 human MASP-1/3 49:62 human MASP-1/3 49:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 3 26 theme -3 501:502 arg1 interaction 475:485 interaction 475:485 interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins 475:571 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 27 with interaction 475:485 arg1 ficolins 564:571 ficolins 564:571 ficolins 564:571 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 27 with interaction 475:485 arg1 proteins 523:530 their partner proteins mannan-binding lectin (MBL) 509:558 their partner proteins mannan-binding lectin (MBL) 509:558 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 0 28 theme sites 102:106 arg1 identification 68:81 identification 68:81 identification 68:81 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 0 28 theme sites 102:106 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 0:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 7 29 theme plasmon 1400:1406 arg1 resonance 1408:1416 surface plasmon resonance 1392:1416 surface plasmon resonance spectroscopy 1392:1429 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 5 30 theme Ca 811:812 arg1 ion 818:820 A Ca(2+) ion 809:820 A Ca(2+) ion bound primarily to both EGF modules 809:856 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 0 31 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 0:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 3 32 theme x-ray 392:396 arg1 structure 398:406 The x-ray structure 388:406 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, 388:572 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 9 33 theme conserved 1788:1796 arg1 lysine 1798:1803 a conserved lysine 1786:1803 a conserved lysine of MBL 1786:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 4 34 theme hydrophobic 678:688 arg1 interactions 690:701 hydrophobic interactions 678:701 hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart 678:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 2 35 theme serine 341:346 arg1 domains 357:363 different serine protease domains 331:363 different serine protease domains at the C-terminal end 331:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 0 36 theme CUB1-EGF-CUB2 25:37 arg1 domain 39:44 the CUB1-EGF-CUB2 domain 21:44 the CUB1-EGF-CUB2 domain of human MASP-1/3 21:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 0 36 theme CUB1-EGF-CUB2 25:37 arg1 MASP-1/3 55:62 human MASP-1/3 49:62 human MASP-1/3 49:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 7 37 theme binding 1353:1359 arg1 properties 1361:1370 their binding properties 1347:1370 their binding properties 1347:1370 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 8 38 from located 1523:1529 arg1 vicinity 1540:1547 close vicinity 1534:1547 close vicinity of their Ca(2+)-binding sites 1534:1577 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 3 39 theme CUB 415:417 arg1 domain 433:438 the CUB(1)-EGF-CUB(2) domain 411:438 the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, 411:572 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 39 theme CUB 415:417 arg1 MASP-1/3 449:456 human MASP-1/3 443:456 human MASP-1/3 443:456 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 6 40 theme Additional 921:930 arg1 2+ 935:936 2+ 935:936 2+ 935:936 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 6 40 theme Additional 921:930 arg1 Ca 932:933 Additional Ca 921:933 Additional Ca(2+) ions 921:942 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 5 41 theme 2+ 814:815 arg1 ion 818:820 A Ca(2+) ion 809:820 A Ca(2+) ion bound primarily to both EGF modules 809:856 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 3 42 theme -EGF-CUB 421:428 arg1 domain 433:438 the CUB(1)-EGF-CUB(2) domain 411:438 the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, 411:572 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 42 theme -EGF-CUB 421:428 arg1 MASP-1/3 449:456 human MASP-1/3 443:456 human MASP-1/3 443:456 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 1 43 theme gene 241:244 arg1 splicing 217:224 alternative splicing 205:224 alternative splicing of the MASP1/3 gene 205:244 MASP-1 and MASP-3 are homologous proteases arising from alternative splicing of the MASP1/3 gene. 18596036 9 44 theme Ca 1755:1756 arg1 ligands 1762:1768 two acidic Ca(2+) ligands 1744:1768 two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL 1744:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 8 45 theme sites 1573:1577 arg1 vicinity 1540:1547 close vicinity 1534:1547 close vicinity of their Ca(2+)-binding sites 1534:1577 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 0 46 with identification 68:81 arg1 ficolins 139:146 ficolins 139:146 ficolins 139:146 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 0 46 with identification 68:81 arg1 lectin 128:133 mannan-binding lectin 113:133 mannan-binding lectin 113:133 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 3 47 theme domain 433:438 arg1 structure 398:406 The x-ray structure 388:406 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, 388:572 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 9 48 theme major 1704:1708 arg1 interaction 1724:1734 a major electrostatic interaction 1702:1734 a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL 1702:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 7 49 theme -3 1259:1260 arg1 H-ficolins 1282:1291 H-ficolins 1282:1291 H-ficolins 1282:1291 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 7 49 theme -3 1259:1260 arg1 interaction 1233:1243 interaction 1233:1243 interaction of MASP-1 and -3 with MBL and L- 1233:1276 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 9 50 theme MASP-1/3 1773:1780 arg1 ligands 1762:1768 two acidic Ca(2+) ligands 1744:1768 two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL 1744:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 7 51 theme MASP-1 1248:1253 arg1 H-ficolins 1282:1291 H-ficolins 1282:1291 H-ficolins 1282:1291 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 7 51 theme MASP-1 1248:1253 arg1 interaction 1233:1243 interaction 1233:1243 interaction of MASP-1 and -3 with MBL and L- 1233:1276 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 10 52 theme other 1832:1836 arg1 data 1838:1841 other data 1832:1841 other data 1832:1841 Based on these and other data, a schematic model of a MBL.MASP complex is proposed. 18596036 3 53 theme human 443:447 arg1 MASP-1/3 449:456 human MASP-1/3 443:456 human MASP-1/3 443:456 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 4 54 theme growth 762:767 arg1 factor 769:774 epidermal growth factor 752:774 the epidermal growth factor (EGF) module of its counterpart 748:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 4 54 theme growth 762:767 arg1 EGF 777:779 EGF 777:779 EGF 777:779 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 9 55 theme lysine 1798:1803 arg1 ligands 1762:1768 two acidic Ca(2+) ligands 1744:1768 two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL 1744:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 10 56 theme schematic 1846:1854 arg1 model 1856:1860 a schematic model 1844:1860 a schematic model of a MBL.MASP complex 1844:1882 Based on these and other data, a schematic model of a MBL.MASP complex is proposed. 18596036 5 57 theme CUB 898:900 arg1 -EGF 904:907 inter-monomer CUB(1)-EGF 884:907 inter-monomer CUB(1)-EGF 884:907 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 5 57 theme CUB 898:900 arg1 1 902:902 1 902:902 1 902:902 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 9 58 theme MBL 1808:1810 arg1 lysine 1798:1803 a conserved lysine 1786:1803 a conserved lysine of MBL 1786:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 9 58 theme MBL 1808:1810 arg1 MASP-1/3 1773:1780 MASP-1/3 1773:1780 MASP-1/3 1773:1780 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 4 59 theme monomer 736:742 arg1 module 722:727 the CUB(1) module 711:727 the CUB(1) module of one monomer 711:742 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 4 59 theme monomer 736:742 arg1 module 782:787 the epidermal growth factor (EGF) module 748:787 the epidermal growth factor (EGF) module of its counterpart 748:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 6 60 theme water 1165:1169 arg1 molecules 1171:1179 one and two water molecules 1153:1179 one and two water molecules 1153:1179 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 2 61 theme identical 263:271 arg1 array 312:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array 260:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end 260:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 8 62 dep modules 1496:1502 arg1 modules 1496:1502 modules CUB(1) and CUB(2) 1496:1520 modules CUB(1) and CUB(2) 1496:1520 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 62 dep modules 1496:1502 arg1 CUB 1515:1517 CUB(2) 1515:1520 CUB(2) 1515:1520 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 62 dep modules 1496:1502 arg1 CUB 1504:1506 CUB(1) 1504:1509 CUB(1) 1504:1509 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 10 63 theme MBL.MASP 1867:1874 arg1 complex 1876:1882 a MBL.MASP complex 1865:1882 a MBL.MASP complex 1865:1882 Based on these and other data, a schematic model of a MBL.MASP complex is proposed. 18596036 6 64 theme Ser 1054:1056 arg1 Glu 1095:1097 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 4 65 theme counterpart 796:806 arg1 module 722:727 the CUB(1) module 711:727 the CUB(1) module of one monomer 711:742 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 4 65 theme counterpart 796:806 arg1 module 782:787 the epidermal growth factor (EGF) module 748:787 the epidermal growth factor (EGF) module of its counterpart 748:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 8 66 theme binding 1467:1473 arg1 sites 1475:1479 two homologous binding sites 1452:1479 two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion 1452:1610 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 67 theme homologous 1456:1465 arg1 sites 1475:1479 two homologous binding sites 1452:1479 two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion 1452:1610 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 2 68 from end 383:385 arg1 domains 357:363 different serine protease domains 331:363 different serine protease domains at the C-terminal end 331:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 0 69 theme interaction 90:100 arg1 sites 102:106 its interaction sites 86:106 its interaction sites 86:106 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 7 70 theme resonance 1408:1416 arg1 spectroscopy 1418:1429 surface plasmon resonance spectroscopy 1392:1429 surface plasmon resonance spectroscopy 1392:1429 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 2 71 theme -EGF-CUB 279:286 arg1 array 312:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array 260:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end 260:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 6 72 theme Asp 1040:1042 arg1 Glu 1095:1097 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 7 73 theme surface 1392:1398 arg1 resonance 1408:1416 surface plasmon resonance 1392:1416 surface plasmon resonance spectroscopy 1392:1429 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 7 74 with interaction 1233:1243 arg1 L- 1275:1276 L- 1275:1276 L- 1275:1276 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 7 74 with interaction 1233:1243 arg1 MBL 1267:1269 MBL 1267:1269 MBL 1267:1269 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 1 75 theme MASP1/3 233:239 arg1 gene 241:244 the MASP1/3 gene 229:244 the MASP1/3 gene 229:244 MASP-1 and MASP-3 are homologous proteases arising from alternative splicing of the MASP1/3 gene. 18596036 5 76 theme -EGF 904:907 arg1 interfaces 909:918 the intra- and inter-monomer CUB(1)-EGF interfaces 869:918 the intra- and inter-monomer CUB(1)-EGF interfaces 869:918 A Ca(2+) ion bound primarily to both EGF modules stabilizes the intra- and inter-monomer CUB(1)-EGF interfaces. 18596036 8 77 theme located 1523:1529 arg1 modules 1496:1502 modules CUB(1) and CUB(2) 1496:1520 modules CUB(1) and CUB(2) 1496:1520 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 77 theme located 1523:1529 arg1 CUB 1515:1517 CUB(2) 1515:1520 CUB(2) 1515:1520 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 77 theme located 1523:1529 arg1 CUB 1504:1506 CUB(1) 1504:1509 CUB(1) 1504:1509 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 2 78 theme C-terminal 372:381 arg1 end 383:385 the C-terminal end 368:385 the C-terminal end 368:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 2 79 theme module 305:310 arg1 array 312:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array 260:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end 260:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 8 80 theme close 1534:1538 arg1 vicinity 1540:1547 close vicinity 1534:1547 close vicinity of their Ca(2+)-binding sites 1534:1577 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 7 81 theme point 1297:1301 arg1 mutants 1303:1309 27 point mutants 1294:1309 27 point mutants of human MASP-3 1294:1325 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 3 82 theme mannan-binding 532:545 arg1 MBL 555:557 MBL 555:557 MBL 555:557 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 82 theme mannan-binding 532:545 arg1 lectin 547:552 mannan-binding lectin 532:552 their partner proteins mannan-binding lectin (MBL) 509:558 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 2 83 theme protease 348:355 arg1 domains 357:363 different serine protease domains 331:363 different serine protease domains at the C-terminal end 331:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 0 84 theme domain 39:44 arg1 identification 68:81 identification 68:81 identification 68:81 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 0 84 theme domain 39:44 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 0:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 3 85 theme partner 515:521 arg1 proteins 523:530 their partner proteins mannan-binding lectin (MBL) 509:558 their partner proteins mannan-binding lectin (MBL) 509:558 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 2 86 theme different 331:339 arg1 domains 357:363 different serine protease domains 331:363 different serine protease domains at the C-terminal end 331:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 0 87 theme human 49:53 arg1 MASP-1/3 55:62 human MASP-1/3 49:62 human MASP-1/3 49:62 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 7 88 theme MASP-3 1320:1325 arg1 mutants 1303:1309 27 point mutants 1294:1309 27 point mutants of human MASP-3 1294:1325 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 6 89 theme Ca 932:933 arg1 ions 939:942 Additional Ca(2+) ions 921:942 Additional Ca(2+) ions 921:942 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 18596036 0 90 with structure 8:16 arg1 ficolins 139:146 ficolins 139:146 ficolins 139:146 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 0 90 with structure 8:16 arg1 lectin 128:133 mannan-binding lectin 113:133 mannan-binding lectin 113:133 Crystal structure of the CUB1-EGF-CUB2 domain of human MASP-1/3 and identification of its interaction sites with mannan-binding lectin and ficolins. 18596036 9 91 theme electrostatic 1710:1722 arg1 interaction 1724:1734 a major electrostatic interaction 1702:1734 a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL 1702:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 7 92 with H-ficolins 1282:1291 arg1 L- 1275:1276 L- 1275:1276 L- 1275:1276 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 7 92 with H-ficolins 1282:1291 arg1 MBL 1267:1269 MBL 1267:1269 MBL 1267:1269 To identify the residues involved in interaction of MASP-1 and -3 with MBL and L- and H-ficolins, 27 point mutants of human MASP-3 were generated, and their binding properties were analyzed using surface plasmon resonance spectroscopy. 18596036 4 93 theme head-to-tail 634:645 arg1 homodimer 647:655 a head-to-tail homodimer 632:655 a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart 632:806 The structure shows a head-to-tail homodimer mainly stabilized by hydrophobic interactions between the CUB(1) module of one monomer and the epidermal growth factor (EGF) module of its counterpart. 18596036 2 94 theme -CCP 290:293 arg1 array 312:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array 260:316 an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end 260:385 They include an identical CUB(1)-EGF-CUB(2)-CCP(1)-CCP(2) module array prolonged by different serine protease domains at the C-terminal end. 18596036 3 95 dep proteins 523:530 arg1 MBL 555:557 MBL 555:557 MBL 555:557 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 3 95 dep proteins 523:530 arg1 lectin 547:552 mannan-binding lectin 532:552 their partner proteins mannan-binding lectin (MBL) 509:558 The x-ray structure of the CUB(1)-EGF-CUB(2) domain of human MASP-1/3, responsible for interaction of MASP-1 and -3 with their partner proteins mannan-binding lectin (MBL) and ficolins, was solved to a resolution of 2.3A(.) 18596036 8 96 from vicinity 1540:1547 arg1 located 1523:1529 located 1523:1529 located 1523:1529 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 8 97 theme -binding 1564:1571 arg1 sites 1573:1577 their Ca(2+)-binding sites 1552:1577 their Ca(2+)-binding sites 1552:1577 These mutations map two homologous binding sites contributed by modules CUB(1) and CUB(2), located in close vicinity of their Ca(2+)-binding sites and stabilized by the Ca(2+) ion. 18596036 9 98 theme acidic 1748:1753 arg1 ligands 1762:1768 two acidic Ca(2+) ligands 1744:1768 two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL 1744:1810 This information allows us to propose a model of the MBL-MASP-1/3 interaction, involving a major electrostatic interaction between two acidic Ca(2+) ligands of MASP-1/3 and a conserved lysine of MBL. 18596036 6 99 theme counterparts 1082:1093 arg1 Glu 1095:1097 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216) 1031:1102 Additional Ca(2+) ions are bound to each CUB(1) and CUB(2) module through six ligands contributed by Glu(49), Asp(57), Asp(102), and Ser(104) (CUB(1)) and their counterparts Glu(216), Asp(226), Asp(263), and Ser(265) (CUB(2)), plus one and two water molecules, respectively. 28554189 6 0 theme nucleotide 1030:1039 arg1 polymorphisms 1041:1053 disease-causing single nucleotide polymorphisms 1007:1053 disease-causing single nucleotide polymorphisms 1007:1053 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 1 theme other 1169:1173 arg1 transporters 1191:1202 other G-subfamily ABC transporters 1169:1202 other G-subfamily ABC transporters 1169:1202 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 4 2 theme inhibitory 604:613 arg1 5D3 624:626 the human-specific, inhibitory antibody 5D3 584:626 5D3 624:626 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 5 3 theme translocation 816:828 arg1 pathway 830:836 a central, hydrophobic, inward-facing translocation pathway 778:836 a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 778:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 5 4 from pathway 830:836 arg1 pocket 752:757 the multidrug-binding pocket 730:757 the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 730:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 5 4 from pathway 830:836 arg1 located 767:773 located 767:773 located 767:773 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 2 5 theme tumour 294:299 arg1 cells 301:305 tumour cells 294:305 tumour cells 294:305 Its activity affects the pharmacokinetics of commonly used drugs and limits the delivery of therapeutics into tumour cells, thus contributing to multidrug resistance. 28554189 4 6 theme 5D3 624:626 arg1 fragments 571:579 two antigen-binding fragments 551:579 two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter 551:681 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 5 7 theme cholesterol 699:709 arg1 molecules 711:719 two cholesterol molecules 695:719 two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 695:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 4 8 theme antibody 615:622 arg1 5D3 624:626 the human-specific, inhibitory antibody 5D3 584:626 5D3 624:626 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 6 9 theme multidrug 939:947 arg1 recognition 949:959 a multidrug recognition and transport mechanism 937:983 recognition 949:959 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 5 10 theme transmembrane 850:862 arg1 domains 864:870 the transmembrane domains 846:870 the transmembrane domains 846:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 2 11 theme therapeutics 276:287 arg1 delivery 264:271 the delivery 260:271 the delivery of therapeutics into tumour cells 260:305 Its activity affects the pharmacokinetics of commonly used drugs and limits the delivery of therapeutics into tumour cells, thus contributing to multidrug resistance. 28554189 1 12 theme xenobiotic 162:171 arg1 molecules 173:181 xenobiotic molecules 162:181 xenobiotic molecules 162:181 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 5 13 theme central 780:786 arg1 pathway 830:836 a central, hydrophobic, inward-facing translocation pathway 778:836 a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 778:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 0 14 theme transporter 33:43 arg1 Structure 0:8 Structure 0:8 Structure of the human multidrug transporter 0:43 Structure of the human multidrug transporter ABCG2. 28554189 3 15 theme first 450:454 arg1 insight 472:478 the first high-resolution insight 446:478 the first high-resolution insight into a human multidrug transporter 446:513 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 3 16 theme human 384:388 arg1 ABCG2 390:394 human ABCG2 384:394 human ABCG2 determined by cryo-electron microscopy 384:433 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 6 17 dep in 898:899 arg1 vitro 901:905 vitro 901:905 vitro 901:905 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 18 theme recognition 1154:1164 arg1 basis 1133:1137 the structural basis 1118:1137 the structural basis of cholesterol recognition by other G-subfamily ABC transporters 1118:1202 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 1 19 theme expressed 78:86 arg1 transporter 115:125 a constitutively expressed ATP-binding cassette (ABC) transporter 61:125 a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules 61:181 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 1 19 theme expressed 78:86 arg1 ABCG2 52:56 ABCG2 52:56 ABCG2 52:56 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 3 20 theme ABCG2 390:394 arg1 structure 371:379 the structure 367:379 the structure of human ABCG2 determined by cryo-electron microscopy 367:433 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 1 21 theme ATP-binding 88:98 arg1 transporter 115:125 a constitutively expressed ATP-binding cassette (ABC) transporter 61:125 a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules 61:181 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 1 21 theme ATP-binding 88:98 arg1 ABCG2 52:56 ABCG2 52:56 ABCG2 52:56 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 0 22 theme human 17:21 arg1 transporter 33:43 the human multidrug transporter 13:43 the human multidrug transporter 13:43 Structure of the human multidrug transporter ABCG2. 28554189 5 23 theme multidrug-binding 734:750 arg1 pocket 752:757 the multidrug-binding pocket 730:757 the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 730:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 5 23 theme multidrug-binding 734:750 arg1 located 767:773 located 767:773 located 767:773 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 1 24 theme cassette 100:107 arg1 transporter 115:125 a constitutively expressed ATP-binding cassette (ABC) transporter 61:125 a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules 61:181 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 1 24 theme cassette 100:107 arg1 ABCG2 52:56 ABCG2 52:56 ABCG2 52:56 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 6 25 theme structural 1122:1131 arg1 basis 1133:1137 the structural basis 1118:1137 the structural basis of cholesterol recognition by other G-subfamily ABC transporters 1118:1202 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 4 26 theme human-specific 588:601 arg1 5D3 624:626 the human-specific, inhibitory antibody 5D3 584:626 5D3 624:626 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 4 27 with complex 538:544 arg1 fragments 571:579 two antigen-binding fragments 551:579 two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter 551:681 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 3 28 theme cryo-electron 410:422 arg1 microscopy 424:433 cryo-electron microscopy 410:433 cryo-electron microscopy 410:433 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 6 29 theme single 1023:1028 arg1 polymorphisms 1041:1053 disease-causing single nucleotide polymorphisms 1007:1053 disease-causing single nucleotide polymorphisms 1007:1053 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 2 30 theme drugs 243:247 arg1 pharmacokinetics 209:224 the pharmacokinetics 205:224 the pharmacokinetics of commonly used drugs 205:247 Its activity affects the pharmacokinetics of commonly used drugs and limits the delivery of therapeutics into tumour cells, thus contributing to multidrug resistance. 28554189 3 31 theme high-resolution 456:470 arg1 insight 472:478 the first high-resolution insight 446:478 the first high-resolution insight into a human multidrug transporter 446:513 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 0 32 theme multidrug 23:31 arg1 transporter 33:43 the human multidrug transporter 13:43 the human multidrug transporter 13:43 Structure of the human multidrug transporter ABCG2. 28554189 6 33 theme disease-causing 1007:1021 arg1 polymorphisms 1041:1053 disease-causing single nucleotide polymorphisms 1007:1053 disease-causing single nucleotide polymorphisms 1007:1053 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 5 34 from located 767:773 arg1 pathway 830:836 a central, hydrophobic, inward-facing translocation pathway 778:836 a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 778:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 2 35 theme used 238:241 arg1 drugs 243:247 commonly used drugs 229:247 commonly used drugs 229:247 Its activity affects the pharmacokinetics of commonly used drugs and limits the delivery of therapeutics into tumour cells, thus contributing to multidrug resistance. 28554189 4 36 theme transporter 671:681 arg1 loops 658:662 extracellular loops 644:662 extracellular loops of the transporter 644:681 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 1 37 theme ABC 110:112 arg1 transporter 115:125 a constitutively expressed ATP-binding cassette (ABC) transporter 61:125 a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules 61:181 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 1 37 theme ABC 110:112 arg1 ABCG2 52:56 ABCG2 52:56 ABCG2 52:56 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 4 38 theme antigen-binding 555:569 arg1 fragments 571:579 two antigen-binding fragments 551:579 two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter 551:681 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 28554189 5 39 dep central 780:786 arg1 hydrophobic 789:799 hydrophobic 789:799 hydrophobic 789:799 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 5 39 dep central 780:786 arg1 inward-facing 802:814 inward-facing 802:814 inward-facing 802:814 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 6 40 theme transport 965:973 arg1 mechanism 975:983 a multidrug recognition and transport mechanism 937:983 mechanism 975:983 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 41 theme ABCG2 988:992 arg1 recognition 949:959 a multidrug recognition and transport mechanism 937:983 recognition 949:959 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 41 theme ABCG2 988:992 arg1 mechanism 975:983 a multidrug recognition and transport mechanism 937:983 mechanism 975:983 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 42 theme in 898:899 arg1 analyses 907:914 functional in vitro analyses 887:914 functional in vitro analyses 887:914 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 2 43 theme multidrug 329:337 arg1 resistance 339:348 multidrug resistance 329:348 multidrug resistance 329:348 Its activity affects the pharmacokinetics of commonly used drugs and limits the delivery of therapeutics into tumour cells, thus contributing to multidrug resistance. 28554189 6 44 theme functional 887:896 arg1 analyses 907:914 functional in vitro analyses 887:914 functional in vitro analyses 887:914 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 3 45 theme multidrug 493:501 arg1 transporter 503:513 a human multidrug transporter 485:513 a human multidrug transporter 485:513 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 3 46 theme human 487:491 arg1 transporter 503:513 a human multidrug transporter 485:513 a human multidrug transporter 485:513 Here we present the structure of human ABCG2 determined by cryo-electron microscopy, providing the first high-resolution insight into a human multidrug transporter. 28554189 6 47 theme G-subfamily 1175:1185 arg1 transporters 1191:1202 other G-subfamily ABC transporters 1169:1202 other G-subfamily ABC transporters 1169:1202 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 5 48 located located 767:773 arg2 located 767:773 located 767:773 located 767:773 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 5 48 located located 767:773 arg2 pocket 752:757 the multidrug-binding pocket 730:757 the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 730:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 5 48 located located 767:773 arg1 pathway 830:836 a central, hydrophobic, inward-facing translocation pathway 778:836 a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains 778:870 We observe two cholesterol molecules bound in the multidrug-binding pocket that is located in a central, hydrophobic, inward-facing translocation pathway between the transmembrane domains. 28554189 6 49 theme ABC 1187:1189 arg1 transporters 1191:1202 other G-subfamily ABC transporters 1169:1202 other G-subfamily ABC transporters 1169:1202 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 50 theme cholesterol 1142:1152 arg1 recognition 1154:1164 cholesterol recognition 1142:1164 cholesterol recognition by other G-subfamily ABC transporters 1142:1202 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 51 theme 5D3 1092:1094 arg1 antibody 1096:1103 the 5D3 antibody 1088:1103 the 5D3 antibody 1088:1103 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 6 52 theme allosteric 1063:1072 arg1 inhibition 1074:1083 the allosteric inhibition 1059:1083 the allosteric inhibition by the 5D3 antibody 1059:1103 Combined with functional in vitro analyses, our results suggest a multidrug recognition and transport mechanism of ABCG2, rationalize disease-causing single nucleotide polymorphisms and the allosteric inhibition by the 5D3 antibody, and provide the structural basis of cholesterol recognition by other G-subfamily ABC transporters. 28554189 1 53 theme many 141:144 arg1 tissues 146:152 many tissues 141:152 many tissues 141:152 ABCG2 is a constitutively expressed ATP-binding cassette (ABC) transporter that protects many tissues against xenobiotic molecules. 28554189 4 54 theme extracellular 644:656 arg1 loops 658:662 extracellular loops 644:662 extracellular loops of the transporter 644:681 We visualize ABCG2 in complex with two antigen-binding fragments of the human-specific, inhibitory antibody 5D3 that recognizes extracellular loops of the transporter. 21768335 3 0 theme therapeutic 539:549 arg1 antibodies 568:577 recombinant therapeutic, glycoenginereed antibodies 527:577 recombinant therapeutic, glycoenginereed antibodies 527:577 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 1 1 theme complexes 264:272 arg1 binding 236:242 the binding 232:242 the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells 232:315 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 6 2 from structure 1085:1093 arg1 weakened 1135:1142 weakened 1135:1142 weakened 1135:1142 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 6 3 theme decreased 1175:1183 arg1 affinity 1185:1192 the decreased affinity 1171:1192 the decreased affinity for the receptor 1171:1209 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 4 4 theme fucosylated 740:750 arg1 Fc 752:753 either afucosylated or fucosylated Fc 717:753 either afucosylated or fucosylated Fc 717:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 5 5 theme receptor 1023:1030 arg1 glycans 1008:1014 glycans 1008:1014 glycans of the receptor and the afucosylated Fc 1008:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 7 6 theme therapeutic 1273:1283 arg1 antibodies 1285:1294 therapeutic antibodies 1273:1294 therapeutic antibodies lacking the core fucose 1273:1318 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 4 7 theme crystal 652:658 arg1 structures 660:669 the crystal structures 648:669 the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc 648:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 6 8 from weakened 1135:1142 arg1 structure 1085:1093 the complex structure 1073:1093 the complex structure with fucosylated Fc 1073:1113 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 6 8 from weakened 1135:1142 arg1 contrast 1060:1067 contrast 1060:1067 contrast 1060:1067 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 5 9 theme Fc 1053:1054 arg1 glycans 1008:1014 glycans 1008:1014 glycans of the receptor and the afucosylated Fc 1008:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 1 10 theme Fcγ 277:279 arg1 receptors 281:289 Fcγ receptors 277:289 Fcγ receptors expressed on immune cells 277:315 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 3 11 dep therapeutic 539:549 arg1 glycoenginereed 552:566 glycoenginereed 552:566 glycoenginereed 552:566 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 5 12 theme carbohydrate-carbohydrate 961:985 arg1 interactions 987:998 carbohydrate-carbohydrate interactions 961:998 carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc 961:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 5 13 dep receptor 1023:1030 arg1 the 1019:1021 the 1019:1021 the 1019:1021 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 4 14 dep detailed 782:789 arg1 molecular 792:800 molecular 792:800 molecular 792:800 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 5 15 gly afucosylated 1040:1051 arg1 Fc 1053:1054 the afucosylated Fc 1036:1054 the afucosylated Fc 1036:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 4 16 theme role 834:837 arg1 understanding 802:814 a detailed, molecular understanding 780:814 a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC 780:895 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 3 17 theme afucosylated 471:482 arg1 IgGs 484:487 afucosylated IgGs 471:487 afucosylated IgGs 471:487 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 0 18 theme core 128:131 arg1 fucose 133:138 core fucose 128:138 core fucose 128:138 Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. 21768335 6 19 theme complex 1077:1083 arg1 structure 1085:1093 the complex structure 1073:1093 the complex structure with fucosylated Fc 1073:1113 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 4 20 theme regulatory 823:832 arg1 role 834:837 the regulatory role 819:837 the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC 819:895 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 1 21 theme key 191:193 arg1 cytotoxicity 168:179 Antibody-mediated cellular cytotoxicity 141:179 Antibody-mediated cellular cytotoxicity (ADCC) 141:186 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 21 theme key 191:193 arg1 mechanism 211:219 a key immune effector mechanism 189:219 a key immune effector mechanism 189:219 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 0 22 theme carbohydrate-carbohydrate 7:31 arg1 interactions 33:44 Unique carbohydrate-carbohydrate interactions 0:44 Unique carbohydrate-carbohydrate interactions 0:44 Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. 21768335 4 23 gly glycosylated 676:687 arg1 receptor 693:700 a glycosylated Fcγ receptor 674:700 a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc 674:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 2 24 from increase 368:375 arg1 affinity 380:387 affinity 380:387 affinity for FcγRIIIa leading to an improved receptor-mediated effector function 380:459 Antibodies lacking core fucosylation show a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function. 21768335 2 25 theme core 337:340 arg1 fucosylation 342:353 core fucosylation 337:353 core fucosylation 337:353 Antibodies lacking core fucosylation show a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function. 21768335 1 26 theme immune 195:200 arg1 cytotoxicity 168:179 Antibody-mediated cellular cytotoxicity 141:179 Antibody-mediated cellular cytotoxicity (ADCC) 141:186 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 26 theme immune 195:200 arg1 mechanism 211:219 a key immune effector mechanism 189:219 a key immune effector mechanism 189:219 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 0 27 theme Unique 0:5 arg1 interactions 33:44 Unique carbohydrate-carbohydrate interactions 0:44 Unique carbohydrate-carbohydrate interactions 0:44 Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. 21768335 4 28 theme detailed 782:789 arg1 understanding 802:814 a detailed, molecular understanding 780:814 a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC 780:895 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 1 29 theme effector 202:209 arg1 cytotoxicity 168:179 Antibody-mediated cellular cytotoxicity 141:179 Antibody-mediated cellular cytotoxicity (ADCC) 141:186 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 29 theme effector 202:209 arg1 mechanism 211:219 a key immune effector mechanism 189:219 a key immune effector mechanism 189:219 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 30 theme immune 304:309 arg1 cells 311:315 immune cells 304:315 immune cells 304:315 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 5 31 theme afucosylated 1040:1051 arg1 Fc 1053:1054 the afucosylated Fc 1036:1054 the afucosylated Fc 1036:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 2 32 theme effector 443:450 arg1 function 452:459 an improved receptor-mediated effector function 413:459 an improved receptor-mediated effector function 413:459 Antibodies lacking core fucosylation show a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function. 21768335 7 33 theme immune 1369:1374 arg1 system 1376:1381 the immune system 1365:1381 the immune system 1365:1381 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 3 34 theme recombinant 527:537 arg1 antibodies 568:577 recombinant therapeutic, glycoenginereed antibodies 527:577 recombinant therapeutic, glycoenginereed antibodies 527:577 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 6 35 from contrast 1060:1067 arg1 weakened 1135:1142 weakened 1135:1142 weakened 1135:1142 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 4 36 theme Fcγ 689:691 arg1 receptor 693:700 a glycosylated Fcγ receptor 674:700 a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc 674:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 5 37 theme interface 937:945 arg1 type 929:932 a unique type 920:932 a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc 920:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 2 38 theme receptor-mediated 425:441 arg1 function 452:459 an improved receptor-mediated effector function 413:459 an improved receptor-mediated effector function 413:459 Antibodies lacking core fucosylation show a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function. 21768335 5 39 theme unique 922:927 arg1 type 929:932 a unique type 920:932 a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc 920:1054 The structures reveal a unique type of interface consisting of carbohydrate-carbohydrate interactions between glycans of the receptor and the afucosylated Fc. 21768335 7 40 theme effector 1414:1421 arg1 functions 1423:1431 antibody-mediated effector functions 1396:1431 antibody-mediated effector functions 1396:1431 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 6 41 with structure 1085:1093 arg1 Fc 1112:1113 fucosylated Fc 1100:1113 fucosylated Fc 1100:1113 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 2 42 theme improved 416:423 arg1 function 452:459 an improved receptor-mediated effector function 413:459 an improved receptor-mediated effector function 413:459 Antibodies lacking core fucosylation show a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function. 21768335 0 43 theme affinity 68:75 arg1 binding 77:83 high affinity binding 63:83 high affinity binding between FcgammaRIII and antibodies lacking core fucose 63:138 Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. 21768335 4 44 gly afucosylated 724:735 arg1 Fc 752:753 either afucosylated or fucosylated Fc 717:753 either afucosylated or fucosylated Fc 717:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 2 45 theme large 362:366 arg1 increase 368:375 a large increase 360:375 a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function 360:459 Antibodies lacking core fucosylation show a large increase in affinity for FcγRIIIa leading to an improved receptor-mediated effector function. 21768335 0 46 theme high 63:66 arg1 binding 77:83 high affinity binding 63:83 high affinity binding between FcgammaRIII and antibodies lacking core fucose 63:138 Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. 21768335 7 47 theme antibody-mediated 1396:1412 arg1 functions 1423:1431 antibody-mediated effector functions 1396:1431 antibody-mediated effector functions 1396:1431 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 7 48 theme antibodies 1285:1294 arg1 efficacy 1261:1268 the higher efficacy 1250:1268 the higher efficacy of therapeutic antibodies lacking the core fucose 1250:1318 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 1 49 theme Antibody-mediated 141:157 arg1 ADCC 182:185 ADCC 182:185 ADCC 182:185 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 49 theme Antibody-mediated 141:157 arg1 cytotoxicity 168:179 Antibody-mediated cellular cytotoxicity 141:179 Antibody-mediated cellular cytotoxicity (ADCC) 141:186 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 49 theme Antibody-mediated 141:157 arg1 mechanism 211:219 a key immune effector mechanism 189:219 a key immune effector mechanism 189:219 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 4 50 gly fucosylated 740:750 arg1 Fc 752:753 either afucosylated or fucosylated Fc 717:753 either afucosylated or fucosylated Fc 717:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 4 51 theme Fc-oligosaccharide 842:859 arg1 fucosylation 866:877 Fc-oligosaccharide core fucosylation 842:877 Fc-oligosaccharide core fucosylation 842:877 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 6 52 theme fucosylated 1100:1110 arg1 Fc 1112:1113 fucosylated Fc 1100:1113 fucosylated Fc 1100:1113 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 1 53 theme cellular 159:166 arg1 ADCC 182:185 ADCC 182:185 ADCC 182:185 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 53 theme cellular 159:166 arg1 cytotoxicity 168:179 Antibody-mediated cellular cytotoxicity 141:179 Antibody-mediated cellular cytotoxicity (ADCC) 141:186 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 1 53 theme cellular 159:166 arg1 mechanism 211:219 a key immune effector mechanism 189:219 a key immune effector mechanism 189:219 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21768335 3 54 gly afucosylated 471:482 arg1 IgGs 484:487 afucosylated IgGs 471:487 afucosylated IgGs 471:487 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 6 55 gly fucosylated 1100:1110 arg1 Fc 1112:1113 fucosylated Fc 1100:1113 fucosylated Fc 1100:1113 In contrast, in the complex structure with fucosylated Fc, these contacts are weakened or nonexistent, explaining the decreased affinity for the receptor. 21768335 4 56 theme afucosylated 724:735 arg1 Fc 752:753 either afucosylated or fucosylated Fc 717:753 either afucosylated or fucosylated Fc 717:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 3 57 theme next 508:511 arg1 generation 513:522 a next generation 506:522 a next generation of recombinant therapeutic, glycoenginereed antibodies 506:577 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 7 58 theme core 1308:1311 arg1 fucose 1313:1318 the core fucose 1304:1318 the core fucose 1304:1318 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 4 59 theme glycosylated 676:687 arg1 receptor 693:700 a glycosylated Fcγ receptor 674:700 a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc 674:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 4 60 theme fucosylation 866:877 arg1 role 834:837 the regulatory role 819:837 the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC 819:895 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 4 61 theme receptor 693:700 arg1 structures 660:669 the crystal structures 648:669 the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc 648:753 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 4 62 theme core 861:864 arg1 fucosylation 866:877 Fc-oligosaccharide core fucosylation 842:877 Fc-oligosaccharide core fucosylation 842:877 In this study, the crystal structures of a glycosylated Fcγ receptor complexed with either afucosylated or fucosylated Fc were determined allowing a detailed, molecular understanding of the regulatory role of Fc-oligosaccharide core fucosylation in improving ADCC. 21768335 7 63 theme unique 1339:1344 arg1 mechanism 1346:1354 a unique mechanism 1337:1354 a unique mechanism by which the immune system can regulate antibody-mediated effector functions 1337:1431 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 3 64 theme antibodies 568:577 arg1 generation 513:522 a next generation 506:522 a next generation of recombinant therapeutic, glycoenginereed antibodies 506:577 Although afucosylated IgGs exist naturally, a next generation of recombinant therapeutic, glycoenginereed antibodies is currently being developed to exploit this finding. 21768335 7 65 theme higher 1254:1259 arg1 efficacy 1261:1268 the higher efficacy 1250:1268 the higher efficacy of therapeutic antibodies lacking the core fucose 1250:1318 These findings allow us to understand the higher efficacy of therapeutic antibodies lacking the core fucose and also suggest a unique mechanism by which the immune system can regulate antibody-mediated effector functions. 21768335 1 66 theme antigen-antibody 247:262 arg1 complexes 264:272 antigen-antibody complexes 247:272 antigen-antibody complexes 247:272 Antibody-mediated cellular cytotoxicity (ADCC), a key immune effector mechanism, relies on the binding of antigen-antibody complexes to Fcγ receptors expressed on immune cells. 21478864 0 0 theme aminopeptidase 89:102 arg1 ERAP1 104:108 the endoplasmic reticulum aminopeptidase ERAP1 63:108 the endoplasmic reticulum aminopeptidase ERAP1 63:108 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 3 1 theme structural 307:316 arg1 basis 318:322 the structural basis 303:322 the structural basis for ERAP1's unusual properties 303:353 To identify the structural basis for ERAP1's unusual properties, we determined the X-ray crystal structure of human ERAP1 bound to bestatin. 21478864 1 2 theme class 158:162 arg1 proteins 166:173 MHC class I proteins 154:173 MHC class I proteins 154:173 ERAP1 trims antigen precursors to fit into MHC class I proteins. 21478864 6 3 theme key 909:911 arg1 residue 923:929 a key catalytic residue 907:929 a key catalytic residue toward the active site 907:952 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 0 4 theme reticulum 79:87 arg1 ERAP1 104:108 the endoplasmic reticulum aminopeptidase ERAP1 63:108 the endoplasmic reticulum aminopeptidase ERAP1 63:108 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 6 5 theme residue 923:929 arg1 reorientation 890:902 reorientation 890:902 reorientation of a key catalytic residue toward the active site 890:952 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 7 6 theme structural 970:979 arg1 elements 981:988 ERAP1's unique structural elements 955:988 ERAP1's unique structural elements 955:988 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 4 7 theme interior 488:495 arg1 compartment 497:507 a large interior compartment 480:507 a large interior compartment 480:507 The structure reveals an open conformation with a large interior compartment. 21478864 5 8 theme broad 644:648 arg1 specificity 650:660 ERAP1's broad specificity 636:660 ERAP1's broad specificity for antigenic peptide precursors 636:693 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 5 9 theme long 592:595 arg1 peptides 597:604 long peptides 592:604 long peptides 592:604 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 4 10 theme open 457:460 arg1 conformation 462:473 an open conformation 454:473 an open conformation with a large interior compartment 454:507 The structure reveals an open conformation with a large interior compartment. 21478864 2 11 theme shorter 277:283 arg1 ones 285:288 shorter ones 277:288 sparing shorter ones 269:288 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 6 12 theme biochemical 711:721 arg1 analyses 723:730 Structural and biochemical analyses 696:730 Structural and biochemical analyses 696:730 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 5 13 contain has 610:612 arg1 groove 522:527 An extended groove 510:527 An extended groove originating from the enzyme's catalytic center 510:574 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 5 13 contain has 610:612 arg2 features 614:621 features 614:621 features that explain ERAP1's broad specificity for antigenic peptide precursors 614:693 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 7 14 theme unique 963:968 arg1 elements 981:988 ERAP1's unique structural elements 955:988 ERAP1's unique structural elements 955:988 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 3 15 theme unusual 336:342 arg1 properties 344:353 ERAP1's unusual properties 328:353 ERAP1's unusual properties 328:353 To identify the structural basis for ERAP1's unusual properties, we determined the X-ray crystal structure of human ERAP1 bound to bestatin. 21478864 3 16 theme X-ray 374:378 arg1 structure 388:396 the X-ray crystal structure 370:396 the X-ray crystal structure of human ERAP1 bound to bestatin 370:429 To identify the structural basis for ERAP1's unusual properties, we determined the X-ray crystal structure of human ERAP1 bound to bestatin. 21478864 6 17 theme Structural 696:705 arg1 analyses 723:730 Structural and biochemical analyses 696:730 Structural and biochemical analyses 696:730 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 7 18 theme precursors 1105:1114 arg1 function 1074:1081 the specialized function 1058:1081 the specialized function of trimming antigenic precursors 1058:1114 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 5 19 theme catalytic 559:567 arg1 center 569:574 the enzyme's catalytic center 546:574 the enzyme's catalytic center 546:574 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 7 20 theme specialized 1062:1072 arg1 function 1074:1081 the specialized function 1058:1081 the specialized function of trimming antigenic precursors 1058:1114 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 3 21 theme ERAP1 407:411 arg1 structure 388:396 the X-ray crystal structure 370:396 the X-ray crystal structure of human ERAP1 bound to bestatin 370:429 To identify the structural basis for ERAP1's unusual properties, we determined the X-ray crystal structure of human ERAP1 bound to bestatin. 21478864 0 22 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 0:109 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 6 23 with change 878:883 arg1 reorientation 890:902 reorientation 890:902 reorientation of a key catalytic residue toward the active site 890:952 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 0 24 theme antigenic 21:29 arg1 processing 49:58 antigenic peptide precursor processing 21:58 antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1 21:108 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 7 25 theme trimming 1086:1093 arg1 precursors 1105:1114 trimming antigenic precursors 1086:1114 trimming antigenic precursors 1086:1114 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 6 26 theme trimming 781:788 arg1 activity 790:797 ERAP1's length-dependent trimming activity 756:797 ERAP1's length-dependent trimming activity 756:797 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 7 27 theme antigenic 1095:1103 arg1 precursors 1105:1114 trimming antigenic precursors 1086:1114 trimming antigenic precursors 1086:1114 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 0 28 theme precursor 39:47 arg1 processing 49:58 antigenic peptide precursor processing 21:58 antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1 21:108 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 3 29 theme human 401:405 arg1 ERAP1 407:411 human ERAP1 401:411 human ERAP1 bound to bestatin 401:429 To identify the structural basis for ERAP1's unusual properties, we determined the X-ray crystal structure of human ERAP1 bound to bestatin. 21478864 0 30 theme peptide 31:37 arg1 processing 49:58 antigenic peptide precursor processing 21:58 antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1 21:108 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 6 31 theme conformational 863:876 arg1 change 878:883 a conformational change 861:883 a conformational change with reorientation of a key catalytic residue toward the active site 861:952 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 2 32 theme substrate 219:227 arg1 preferences 229:239 unique substrate preferences 212:239 unique substrate preferences 212:239 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 6 33 theme length-dependent 764:779 arg1 activity 790:797 ERAP1's length-dependent trimming activity 756:797 ERAP1's length-dependent trimming activity 756:797 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 3 34 theme crystal 380:386 arg1 structure 388:396 the X-ray crystal structure 370:396 the X-ray crystal structure of human ERAP1 bound to bestatin 370:429 To identify the structural basis for ERAP1's unusual properties, we determined the X-ray crystal structure of human ERAP1 bound to bestatin. 21478864 2 35 theme unique 212:217 arg1 preferences 229:239 unique substrate preferences 212:239 unique substrate preferences 212:239 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 1 36 theme antigen 123:129 arg1 precursors 131:140 antigen precursors 123:140 antigen precursors 123:140 ERAP1 trims antigen precursors to fit into MHC class I proteins. 21478864 6 37 theme catalytic 913:921 arg1 residue 923:929 a key catalytic residue 907:929 a key catalytic residue toward the active site 907:952 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 5 38 theme antigenic 666:674 arg1 precursors 684:693 antigenic peptide precursors 666:693 antigenic peptide precursors 666:693 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 7 39 theme aminopeptidase 1012:1025 arg1 structure 1027:1035 a generic aminopeptidase structure 1002:1035 a generic aminopeptidase structure 1002:1035 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 6 40 theme active 942:947 arg1 site 949:952 the active site 938:952 the active site 938:952 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 5 41 theme peptide 676:682 arg1 precursors 684:693 antigenic peptide precursors 666:693 antigenic peptide precursors 666:693 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 6 42 theme substrates 842:851 arg1 binding 808:814 binding 808:814 binding of long rather than short substrates 808:851 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 6 43 theme long 819:822 arg1 substrates 842:851 long rather than short substrates 819:851 long rather than short substrates 819:851 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 2 44 contain has 208:210 arg2 preferences 229:239 unique substrate preferences 212:239 unique substrate preferences 212:239 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 2 44 contain has 208:210 arg1 ERAP1 202:206 ERAP1 202:206 ERAP1 202:206 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 6 45 theme short 836:840 arg1 substrates 842:851 long rather than short substrates 819:851 long rather than short substrates 819:851 Structural and biochemical analyses suggest a mechanism for ERAP1's length-dependent trimming activity, whereby binding of long rather than short substrates induces a conformational change with reorientation of a key catalytic residue toward the active site. 21478864 2 46 theme long 251:254 arg1 peptides 256:263 long peptides 251:263 long peptides 251:263 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 4 47 with conformation 462:473 arg1 compartment 497:507 a large interior compartment 480:507 a large interior compartment 480:507 The structure reveals an open conformation with a large interior compartment. 21478864 0 48 theme endoplasmic 67:77 arg1 ERAP1 104:108 the endoplasmic reticulum aminopeptidase ERAP1 63:108 the endoplasmic reticulum aminopeptidase ERAP1 63:108 Structural basis for antigenic peptide precursor processing by the endoplasmic reticulum aminopeptidase ERAP1. 21478864 7 49 theme generic 1004:1010 arg1 structure 1027:1035 a generic aminopeptidase structure 1002:1035 a generic aminopeptidase structure 1002:1035 ERAP1's unique structural elements suggest how a generic aminopeptidase structure has been adapted for the specialized function of trimming antigenic precursors. 21478864 2 50 dep sparing 269:275 arg1 ones 285:288 shorter ones 277:288 sparing shorter ones 269:288 To fulfill this function, ERAP1 has unique substrate preferences, trimming long peptides but sparing shorter ones. 21478864 4 51 theme large 482:486 arg1 compartment 497:507 a large interior compartment 480:507 a large interior compartment 480:507 The structure reveals an open conformation with a large interior compartment. 21478864 5 52 theme extended 513:520 arg1 groove 522:527 An extended groove 510:527 An extended groove originating from the enzyme's catalytic center 510:574 An extended groove originating from the enzyme's catalytic center can accommodate long peptides and has features that explain ERAP1's broad specificity for antigenic peptide precursors. 21478864 1 53 theme MHC 154:156 arg1 class 158:162 MHC class I 154:164 MHC class I proteins 154:173 ERAP1 trims antigen precursors to fit into MHC class I proteins. 22052913 0 0 theme alternate 100:108 arg1 modes 110:114 alternate modes 100:114 alternate modes of binding 100:125 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 10 1 theme binding 1358:1364 arg1 modes 1342:1346 distinctive modes 1330:1346 distinctive modes 1330:1346 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 8 2 theme receptor-binding 1073:1088 arg1 site 1090:1093 the type I receptor-binding site 1062:1093 the type I receptor-binding site of the ligand 1062:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 8 2 theme receptor-binding 1073:1088 arg1 ligand 1102:1107 the ligand 1098:1107 the ligand 1098:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 10 3 theme ligand 1351:1356 arg1 binding 1358:1364 ligand binding 1351:1364 ligand binding 1351:1364 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 9 4 theme Fstl3 1164:1168 arg1 ND 1158:1159 the ND 1154:1159 the ND of Fstl3 1154:1168 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 9 5 theme corresponding 1213:1225 arg1 mutations 1227:1235 corresponding mutations 1213:1235 corresponding mutations in Fst 1213:1242 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 5 6 theme Fst-type 643:650 arg1 3 614:614 3 614:614 3 614:614 Myostatin is tightly controlled by Fst-like 3 (Fstl3), which is the only Fst-type molecule that has been identified in the serum bound to myostatin. 22052913 5 6 theme Fst-type 643:650 arg1 molecule 652:659 the only Fst-type molecule 634:659 the only Fst-type molecule that has been identified in the serum bound to myostatin 634:716 Myostatin is tightly controlled by Fst-like 3 (Fstl3), which is the only Fst-type molecule that has been identified in the serum bound to myostatin. 22052913 7 7 from surfaces 949:956 arg1 ligand 965:970 the ligand 961:970 the ligand 961:970 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 2 8 theme growth 295:300 arg1 regulator 275:283 a staunch negative regulator 256:283 a staunch negative regulator of muscle growth 256:300 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 8 theme growth 295:300 arg1 myostatin 243:251 the TGF-β ligand myostatin 226:251 the TGF-β ligand myostatin 226:251 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 8 theme growth 295:300 arg1 target 320:325 a therapeutic target 306:325 a therapeutic target for muscle-wasting disorders 306:354 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 8 9 from position 1050:1057 arg1 site 1090:1093 the type I receptor-binding site 1062:1093 the type I receptor-binding site of the ligand 1062:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 8 9 from position 1050:1057 arg1 ligand 1102:1107 the ligand 1098:1107 the ligand 1098:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 7 10 theme activin 908:914 arg1 A 916:916 activin A 908:916 activin A 908:916 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 2 11 theme muscle 288:293 arg1 growth 295:300 muscle growth 288:300 muscle growth 288:300 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 7 12 theme N-terminal 825:834 arg1 ND 844:845 ND 844:845 ND 844:845 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 7 12 theme N-terminal 825:834 arg1 Fstl3 851:855 Fstl3 851:855 Fstl3 851:855 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 7 12 theme N-terminal 825:834 arg1 domain 836:841 the N-terminal domain 821:841 the N-terminal domain (ND) of Fstl3 821:855 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 8 13 theme type 1066:1069 arg1 site 1090:1093 the type I receptor-binding site 1062:1093 the type I receptor-binding site of the ligand 1062:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 8 13 theme type 1066:1069 arg1 ligand 1102:1107 the ligand 1098:1107 the ligand 1098:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 8 14 theme conformational 989:1002 arg1 differences 1004:1014 conformational differences 989:1014 conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand 989:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 9 15 from mutations 1141:1149 arg1 ND 1158:1159 the ND 1154:1159 the ND of Fstl3 1154:1168 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 1 16 theme physiological 187:199 arg1 processes 201:209 critical physiological processes 178:209 critical physiological processes 178:209 TGF-β family ligands are involved in a variety of critical physiological processes. 22052913 0 17 theme binding 119:125 arg1 modes 110:114 alternate modes 100:114 alternate modes of binding 100:125 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 4 18 theme extracellular 499:511 arg1 proteins 560:567 the follistatin (Fst)-type proteins 533:567 the follistatin (Fst)-type proteins 533:567 One form of regulation is through inhibition by extracellular antagonists such as the follistatin (Fst)-type proteins. 22052913 4 18 theme extracellular 499:511 arg1 antagonists 513:523 extracellular antagonists 499:523 extracellular antagonists such as the follistatin (Fst)-type proteins 499:567 One form of regulation is through inhibition by extracellular antagonists such as the follistatin (Fst)-type proteins. 22052913 6 19 theme myostatin 761:769 arg1 structure 748:756 the crystal structure 736:756 the crystal structure of myostatin in complex with Fstl3 736:791 Here, we present the crystal structure of myostatin in complex with Fstl3. 22052913 1 20 theme critical 178:185 arg1 processes 201:209 critical physiological processes 178:209 critical physiological processes 178:209 TGF-β family ligands are involved in a variety of critical physiological processes. 22052913 6 21 with complex 774:780 arg1 Fstl3 787:791 Fstl3 787:791 Fstl3 787:791 Here, we present the crystal structure of myostatin in complex with Fstl3. 22052913 10 22 theme distinctive 1330:1340 arg1 modes 1342:1346 distinctive modes 1330:1346 distinctive modes 1330:1346 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 9 23 theme point 1135:1139 arg1 mutations 1141:1149 single point mutations 1128:1149 single point mutations in the ND of Fstl3 1128:1168 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 3 24 theme family 432:437 arg1 regulation 439:448 TGF-β family regulation 426:448 TGF-β family regulation 426:448 Therefore, it is important to understand the molecular mechanisms of TGF-β family regulation. 22052913 9 25 theme little 1249:1254 arg1 effect 1256:1261 little effect 1249:1261 little effect 1249:1261 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 10 26 theme molecules 1312:1320 arg1 NDs 1296:1298 the NDs 1292:1298 the NDs of Fst-type molecules 1292:1320 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 2 27 theme therapeutic 308:318 arg1 target 320:325 a therapeutic target 306:325 a therapeutic target for muscle-wasting disorders 306:354 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 9 28 contain have 1244:1247 arg2 effect 1256:1261 little effect 1249:1261 little effect 1249:1261 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 9 28 contain have 1244:1247 arg1 mutations 1227:1235 corresponding mutations 1213:1235 corresponding mutations in Fst 1213:1242 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 3 29 theme regulation 439:448 arg1 mechanisms 412:421 the molecular mechanisms 398:421 the molecular mechanisms of TGF-β family regulation 398:448 Therefore, it is important to understand the molecular mechanisms of TGF-β family regulation. 22052913 9 30 from mutations 1227:1235 arg1 Fst 1240:1242 Fst 1240:1242 Fst 1240:1242 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 4 31 theme -type 554:558 arg1 proteins 560:567 the follistatin (Fst)-type proteins 533:567 the follistatin (Fst)-type proteins 533:567 One form of regulation is through inhibition by extracellular antagonists such as the follistatin (Fst)-type proteins. 22052913 8 32 theme Fstl3 1029:1033 arg1 ND 1023:1024 the ND 1019:1024 the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand 1019:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 6 33 theme crystal 740:746 arg1 structure 748:756 the crystal structure 736:756 the crystal structure of myostatin in complex with Fstl3 736:791 Here, we present the crystal structure of myostatin in complex with Fstl3. 22052913 6 34 from structure 748:756 arg1 complex 774:780 complex 774:780 complex with Fstl3 774:791 Here, we present the crystal structure of myostatin in complex with Fstl3. 22052913 0 35 theme myostatin·follistatin-like 13:38 arg1 Structure 0:8 Structure 0:8 Structure of myostatin·follistatin-like 3 0:40 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 0 36 theme N-terminal 43:52 arg1 domains 54:60 N-terminal domains 43:60 N-terminal domains of follistatin-type molecules 43:90 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 0 36 theme N-terminal 43:52 arg1 molecules 82:90 follistatin-type molecules 65:90 follistatin-type molecules 65:90 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 1 37 theme processes 201:209 arg1 processes 201:209 critical physiological processes 178:209 critical physiological processes 178:209 TGF-β family ligands are involved in a variety of critical physiological processes. 22052913 1 37 theme processes 201:209 arg1 variety 167:173 a variety 165:173 a variety of critical physiological processes 165:209 TGF-β family ligands are involved in a variety of critical physiological processes. 22052913 3 38 theme TGF-β 426:430 arg1 regulation 439:448 TGF-β family regulation 426:448 TGF-β family regulation 426:448 Therefore, it is important to understand the molecular mechanisms of TGF-β family regulation. 22052913 10 39 theme Fst-type 1303:1310 arg1 molecules 1312:1320 Fst-type molecules 1303:1320 Fst-type molecules 1303:1320 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 10 40 theme overall 1384:1390 arg1 affinity 1392:1399 overall affinity 1384:1399 overall affinity of ligand·Fst-type protein complexes 1384:1436 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 2 41 theme ligand 236:241 arg1 regulator 275:283 a staunch negative regulator 256:283 a staunch negative regulator of muscle growth 256:300 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 41 theme ligand 236:241 arg1 myostatin 243:251 the TGF-β ligand myostatin 226:251 the TGF-β ligand myostatin 226:251 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 8 42 theme ligand 1102:1107 arg1 site 1090:1093 the type I receptor-binding site 1062:1093 the type I receptor-binding site of the ligand 1062:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 8 42 theme ligand 1102:1107 arg1 ligand 1102:1107 the ligand 1098:1107 the ligand 1098:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 3 43 theme molecular 402:410 arg1 mechanisms 412:421 the molecular mechanisms 398:421 the molecular mechanisms of TGF-β family regulation 398:448 Therefore, it is important to understand the molecular mechanisms of TGF-β family regulation. 22052913 4 44 theme follistatin 537:547 arg1 proteins 560:567 the follistatin (Fst)-type proteins 533:567 the follistatin (Fst)-type proteins 533:567 One form of regulation is through inhibition by extracellular antagonists such as the follistatin (Fst)-type proteins. 22052913 4 45 theme regulation 463:472 arg1 form 455:458 One form 451:458 One form of regulation 451:472 One form of regulation is through inhibition by extracellular antagonists such as the follistatin (Fst)-type proteins. 22052913 2 46 theme TGF-β 230:234 arg1 regulator 275:283 a staunch negative regulator 256:283 a staunch negative regulator of muscle growth 256:300 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 46 theme TGF-β 230:234 arg1 myostatin 243:251 the TGF-β ligand myostatin 226:251 the TGF-β ligand myostatin 226:251 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 1 47 theme TGF-β 128:132 arg1 ligands 141:147 TGF-β family ligands 128:147 TGF-β family ligands 128:147 TGF-β family ligands are involved in a variety of critical physiological processes. 22052913 10 48 theme ligand·Fst-type 1404:1418 arg1 complexes 1428:1436 ligand·Fst-type protein complexes 1404:1436 ligand·Fst-type protein complexes 1404:1436 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 1 49 theme family 134:139 arg1 ligands 141:147 TGF-β family ligands 128:147 TGF-β family ligands 128:147 TGF-β family ligands are involved in a variety of critical physiological processes. 22052913 8 50 from differences 1004:1014 arg1 ND 1023:1024 the ND 1019:1024 the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand 1019:1107 This results in conformational differences in the ND of Fstl3 that alter its position in the type I receptor-binding site of the ligand. 22052913 10 51 theme complexes 1428:1436 arg1 affinity 1392:1399 overall affinity 1384:1399 overall affinity of ligand·Fst-type protein complexes 1384:1436 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 9 52 theme single 1128:1133 arg1 mutations 1141:1149 single point mutations 1128:1149 single point mutations in the ND of Fstl3 1128:1168 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 2 53 theme negative 266:273 arg1 regulator 275:283 a staunch negative regulator 256:283 a staunch negative regulator of muscle growth 256:300 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 53 theme negative 266:273 arg1 myostatin 243:251 the TGF-β ligand myostatin 226:251 the TGF-β ligand myostatin 226:251 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 54 theme muscle-wasting 331:344 arg1 disorders 346:354 muscle-wasting disorders 331:354 muscle-wasting disorders 331:354 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 10 55 theme protein 1420:1426 arg1 complexes 1428:1436 ligand·Fst-type protein complexes 1404:1436 ligand·Fst-type protein complexes 1404:1436 Overall, we have shown that the NDs of Fst-type molecules exhibit distinctive modes of ligand binding, which may affect overall affinity of ligand·Fst-type protein complexes. 22052913 2 56 theme staunch 258:264 arg1 regulator 275:283 a staunch negative regulator 256:283 a staunch negative regulator of muscle growth 256:300 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 2 56 theme staunch 258:264 arg1 myostatin 243:251 the TGF-β ligand myostatin 226:251 the TGF-β ligand myostatin 226:251 For instance, the TGF-β ligand myostatin is a staunch negative regulator of muscle growth and a therapeutic target for muscle-wasting disorders. 22052913 7 57 theme different 939:947 arg1 surfaces 949:956 different surfaces 939:956 different surfaces on the ligand 939:970 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 7 58 theme Fstl3 851:855 arg1 ND 844:845 ND 844:845 ND 844:845 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 7 58 theme Fstl3 851:855 arg1 Fstl3 851:855 Fstl3 851:855 Fstl3 851:855 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 7 58 theme Fstl3 851:855 arg1 domain 836:841 the N-terminal domain 821:841 the N-terminal domain (ND) of Fstl3 821:855 The structure reveals that the N-terminal domain (ND) of Fstl3 interacts uniquely with myostatin as compared with activin A, because it utilizes different surfaces on the ligand. 22052913 0 59 theme molecules 82:90 arg1 domains 54:60 N-terminal domains 43:60 N-terminal domains of follistatin-type molecules 43:90 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 0 59 theme molecules 82:90 arg1 molecules 82:90 follistatin-type molecules 65:90 follistatin-type molecules 65:90 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 22052913 9 60 theme ligand 1189:1194 arg1 binding 1196:1202 ligand binding 1189:1202 ligand binding 1189:1202 We also show that single point mutations in the ND of Fstl3 are detrimental to ligand binding, whereas corresponding mutations in Fst have little effect. 22052913 0 61 theme follistatin-type 65:80 arg1 molecules 82:90 follistatin-type molecules 65:90 follistatin-type molecules 65:90 Structure of myostatin·follistatin-like 3: N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. 1544894 3 0 theme protein 620:626 arg1 C 628:628 protein C 620:628 protein C 620:628 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 4 1 theme mass 818:821 arg1 analysis 837:844 mass spectrometric analysis 818:844 mass spectrometric analysis 818:844 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 7 2 theme VII 1141:1143 arg1 VII 1141:1143 human factor VII 1128:1143 human factor VII 1128:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 2 theme VII 1141:1143 arg1 domain 1118:1123 the first EGF domain 1104:1123 the first EGF domain of human factor VII 1104:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 3 theme factor 1134:1139 arg1 VII 1141:1143 human factor VII 1128:1143 human factor VII 1128:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 3 4 from position 664:671 arg1 residues 646:653 3 residues 644:653 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71 644:744 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 0 5 theme XII 81:83 arg1 domain 64:69 the first epidermal growth factor domain 30:69 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 5 theme XII 81:83 arg1 XII 81:83 factor XII 74:83 factor XII 74:83 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 10 6 theme exclusive 1591:1599 arg1 modifications 1620:1632 mutually exclusive post-translational modifications 1582:1632 mutually exclusive post-translational modifications 1582:1632 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 6 theme exclusive 1591:1599 arg1 fucosylation 1527:1538 O-linked fucosylation 1518:1538 O-linked fucosylation 1518:1538 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 6 theme exclusive 1591:1599 arg1 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 1 7 theme EGF 406:408 arg1 domains 410:416 their EGF domains 400:416 their EGF domains 400:416 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 0 8 theme factor 74:79 arg1 XII 81:83 factor XII 74:83 factor XII 74:83 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 2 9 theme N 519:519 arg1 terminus 521:528 the N terminus 515:528 the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA 515:614 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 0 10 theme coagulation/fibrinolytic 196:219 arg1 system 221:226 the coagulation/fibrinolytic system 192:226 the coagulation/fibrinolytic system 192:226 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 3 11 theme Asp-71 739:744 arg1 beta-hydroxylation 717:734 beta-hydroxylation 717:734 beta-hydroxylation of Asp-71 717:744 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 3 11 theme Asp-71 739:744 arg1 modification 703:714 another post-translational modification 676:714 another post-translational modification 676:714 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 0 12 link O-linked 0:7 arg1 fucose 9:14 O-linked fucose 0:14 O-linked fucose 0:14 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 2 13 theme homologous 568:577 arg1 position 559:566 a position 557:566 a position homologous to that which is fucosylated in t-PA 557:614 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 2 14 theme Factor 419:424 arg1 XII 426:428 Factor XII 419:428 Factor XII 419:428 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 0 15 theme many 155:158 arg1 proteins 160:167 many proteins 155:167 many proteins 155:167 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 15 theme many 155:158 arg1 those 183:187 those 183:187 those 183:187 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 7 16 theme first 1108:1112 arg1 VII 1141:1143 human factor VII 1128:1143 human factor VII 1128:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 16 theme first 1108:1112 arg1 domain 1118:1123 the first EGF domain 1104:1123 the first EGF domain of human factor VII 1104:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 1 17 attach attachment 339:348 arg1 residues 384:391 equivalent threonine residues 363:391 equivalent threonine residues within their EGF domains 363:416 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 1 17 attach attachment 339:348 arg2 fucose 353:358 fucose 353:358 fucose 353:358 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 5 18 gly fucosylated 943:953 arg1 XII 930:932 factor XII 923:932 factor XII 923:932 We found that factor XII is fully fucosylated at Thr-90. 1544894 5 18 gly fucosylated 943:953 arg2 Thr-90 958:963 Thr-90 958:963 Thr-90 958:963 We found that factor XII is fully fucosylated at Thr-90. 1544894 2 19 dep terminus 521:528 arg1 has 530:532 has 530:532 has a threonine residue in a position homologous to that which is fucosylated in t-PA 530:614 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 7 20 theme human 1128:1132 arg1 VII 1141:1143 human factor VII 1128:1143 human factor VII 1128:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 4 21 theme O-linked 863:870 arg1 fucose 872:877 O-linked fucose 863:877 O-linked fucose 863:877 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 9 22 theme consensus 1410:1418 arg1 sequence 1420:1427 the consensus sequence 1406:1427 the consensus sequence for beta-hydroxylation 1406:1450 266, 11051-11057), while it is unmodified at Asp-63 despite having the consensus sequence for beta-hydroxylation at the latter site. 1544894 9 23 dep 266 1339:1341 arg1 11051-11057 1344:1354 11051-11057 1344:1354 11051-11057 1344:1354 266, 11051-11057), while it is unmodified at Asp-63 despite having the consensus sequence for beta-hydroxylation at the latter site. 1544894 9 23 dep 266 1339:1341 arg1 while 1358:1362 while 1358:1362 while 1358:1362 266, 11051-11057), while it is unmodified at Asp-63 despite having the consensus sequence for beta-hydroxylation at the latter site. 1544894 0 24 theme O-linked 0:7 arg1 fucose 9:14 O-linked fucose 0:14 O-linked fucose 0:14 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 10 25 gly fucosylation 1527:1538 arg1 domains 1570:1576 EGF domains 1566:1576 EGF domains 1566:1576 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 2 26 theme threonine 536:544 arg1 residue 546:552 a threonine residue 534:552 a threonine residue 534:552 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 7 27 theme EGF 1114:1116 arg1 VII 1141:1143 human factor VII 1128:1143 human factor VII 1128:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 27 theme EGF 1114:1116 arg1 domain 1118:1123 the first EGF domain 1104:1123 the first EGF domain of human factor VII 1104:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 0 28 theme C. 101:102 arg1 domains 134:140 protein C. Epidermal growth factor (EGF) domains 93:140 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 10 29 link O-linked 1518:1525 arg1 modifications 1620:1632 mutually exclusive post-translational modifications 1582:1632 mutually exclusive post-translational modifications 1582:1632 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 29 link O-linked 1518:1525 arg1 fucosylation 1527:1538 O-linked fucosylation 1518:1538 O-linked fucosylation 1518:1538 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 29 link O-linked 1518:1525 arg1 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 0 30 theme protein 93:99 arg1 domains 134:140 protein C. Epidermal growth factor (EGF) domains 93:140 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 2 31 theme protein 434:440 arg1 C 442:442 protein C 434:442 protein C 434:442 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 0 32 theme growth 114:119 arg1 EGF 129:131 EGF 129:131 EGF 129:131 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 32 theme growth 114:119 arg1 factor 121:126 Epidermal growth factor 104:126 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 10 33 theme domains 1570:1576 arg1 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 33 theme domains 1570:1576 arg1 fucosylation 1527:1538 O-linked fucosylation 1518:1538 O-linked fucosylation 1518:1538 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 33 theme domains 1570:1576 arg1 modifications 1620:1632 mutually exclusive post-translational modifications 1582:1632 mutually exclusive post-translational modifications 1582:1632 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 6 34 theme Protein 966:972 arg1 C 974:974 Protein C 966:974 Protein C 966:974 Protein C is unmodified at the equivalent site (Thr-68) and is completely beta-hydroxylated at Asp-71. 1544894 6 35 theme equivalent 997:1006 arg1 Thr-68 1014:1019 Thr-68 1014:1019 Thr-68 1014:1019 Protein C is unmodified at the equivalent site (Thr-68) and is completely beta-hydroxylated at Asp-71. 1544894 6 35 theme equivalent 997:1006 arg1 site 1008:1011 the equivalent site 993:1011 the equivalent site (Thr-68) 993:1020 Protein C is unmodified at the equivalent site (Thr-68) and is completely beta-hydroxylated at Asp-71. 1544894 1 36 theme tissue 266:271 arg1 t-PA 296:299 t-PA 296:299 t-PA 296:299 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 1 36 theme tissue 266:271 arg1 activator 285:293 tissue plasminogen activator 266:293 tissue plasminogen activator (t-PA) 266:300 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 0 37 theme Epidermal 104:112 arg1 EGF 129:131 EGF 129:131 EGF 129:131 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 37 theme Epidermal 104:112 arg1 factor 121:126 Epidermal growth factor 104:126 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 1 38 theme fucose 353:358 arg1 attachment 339:348 the attachment 335:348 the attachment of fucose to equivalent threonine residues within their EGF domains 335:416 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 10 39 theme EGF 1566:1568 arg1 domains 1570:1576 EGF domains 1566:1576 EGF domains 1566:1576 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 2 40 theme EGF 461:463 arg1 domains 465:471 two EGF domains 457:471 two EGF domains 457:471 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 0 41 theme epidermal 40:48 arg1 factor 57:62 the first epidermal growth factor 30:62 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 1 42 theme plasminogen 273:283 arg1 t-PA 296:299 t-PA 296:299 t-PA 296:299 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 1 42 theme plasminogen 273:283 arg1 activator 285:293 tissue plasminogen activator 266:293 tissue plasminogen activator (t-PA) 266:300 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 9 43 theme latter 1459:1464 arg1 site 1466:1469 the latter site 1455:1469 the latter site 1455:1469 266, 11051-11057), while it is unmodified at Asp-63 despite having the consensus sequence for beta-hydroxylation at the latter site. 1544894 4 44 theme beta-hydroxyaspartate 886:906 arg1 presence 851:858 the presence 847:858 the presence of O-linked fucose and/or beta-hydroxyaspartate 847:906 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 4 45 theme spectrometric 823:835 arg1 analysis 837:844 mass spectrometric analysis 818:844 mass spectrometric analysis 818:844 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 0 46 theme first 34:38 arg1 factor 57:62 the first epidermal growth factor 30:62 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 47 theme factor 121:126 arg1 domains 134:140 protein C. Epidermal growth factor (EGF) domains 93:140 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 1 48 theme equivalent 363:372 arg1 residues 384:391 equivalent threonine residues 363:391 equivalent threonine residues within their EGF domains 363:416 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 2 49 contain contain 449:455 arg1 XII 426:428 Factor XII 419:428 Factor XII 419:428 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 2 49 contain contain 449:455 arg2 domains 465:471 two EGF domains 457:471 two EGF domains 457:471 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 2 49 contain contain 449:455 arg1 C 442:442 protein C 434:442 protein C 434:442 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 4 50 contain containing 768:777 arg1 peptides 759:766 peptides 759:766 peptides containing these sites 759:789 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 4 50 contain containing 768:777 arg2 sites 785:789 these sites 779:789 these sites 779:789 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 4 51 link O-linked 863:870 arg1 fucose 872:877 O-linked fucose 863:877 O-linked fucose 863:877 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 1 52 theme threonine 374:382 arg1 residues 384:391 equivalent threonine residues 363:391 equivalent threonine residues within their EGF domains 363:416 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 10 53 theme post-translational 1601:1618 arg1 modifications 1620:1632 mutually exclusive post-translational modifications 1582:1632 mutually exclusive post-translational modifications 1582:1632 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 53 theme post-translational 1601:1618 arg1 fucosylation 1527:1538 O-linked fucosylation 1518:1538 O-linked fucosylation 1518:1538 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 53 theme post-translational 1601:1618 arg1 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 7 54 dep Christensen 1254:1264 arg1 Biol 1327:1330 Biol 1327:1330 Biol 1327:1330 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 10 55 theme O-linked 1518:1525 arg1 modifications 1620:1632 mutually exclusive post-translational modifications 1582:1632 mutually exclusive post-translational modifications 1582:1632 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 55 theme O-linked 1518:1525 arg1 fucosylation 1527:1538 O-linked fucosylation 1518:1538 O-linked fucosylation 1518:1538 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 10 55 theme O-linked 1518:1525 arg1 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 beta-hydroxylation 1544:1561 These observations raise the possibility that O-linked fucosylation and beta-hydroxylation of EGF domains are mutually exclusive post-translational modifications. 1544894 2 56 gly fucosylated 596:606 arg1 that 582:585 that 582:585 that 582:585 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 2 57 theme EGF 496:498 arg1 domain 500:505 the EGF domain 492:505 the EGF domain 492:505 Factor XII and protein C each contain two EGF domains; in both proteins, the EGF domain nearest the N terminus has a threonine residue in a position homologous to that which is fucosylated in t-PA. 1544894 1 58 mod modified 323:330 arg3 attachment 339:348 the attachment 335:348 the attachment of fucose to equivalent threonine residues within their EGF domains 335:416 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 1 58 mod modified 323:330 arg1 t-PA 296:299 t-PA 296:299 t-PA 296:299 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 1 58 mod modified 323:330 arg1 prourokinase 306:317 prourokinase 306:317 prourokinase 306:317 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 1 58 mod modified 323:330 arg1 activator 285:293 tissue plasminogen activator 266:293 tissue plasminogen activator (t-PA) 266:300 We and others have demonstrated that tissue plasminogen activator (t-PA) and prourokinase are modified by the attachment of fucose to equivalent threonine residues within their EGF domains. 1544894 5 59 theme factor 923:928 arg1 XII 930:932 factor XII 923:932 factor XII 923:932 We found that factor XII is fully fucosylated at Thr-90. 1544894 0 60 located found 146:150 arg1 proteins 160:167 many proteins 155:167 many proteins 155:167 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 60 located found 146:150 arg1 those 183:187 those 183:187 those 183:187 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 60 located found 146:150 arg2 domain 64:69 the first epidermal growth factor domain 30:69 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 60 located found 146:150 arg2 XII 81:83 factor XII 74:83 factor XII 74:83 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 60 located found 146:150 arg2 domains 134:140 protein C. Epidermal growth factor (EGF) domains 93:140 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 61 theme factor 57:62 arg1 domain 64:69 the first epidermal growth factor domain 30:69 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 0 61 theme factor 57:62 arg1 XII 81:83 factor XII 74:83 factor XII 74:83 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 3 62 from residues 646:653 arg1 C 628:628 protein C 620:628 protein C 620:628 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 9 63 from Asp-63 1384:1389 arg1 unmodified 1370:1379 unmodified 1370:1379 unmodified 1370:1379 266, 11051-11057), while it is unmodified at Asp-63 despite having the consensus sequence for beta-hydroxylation at the latter site. 1544894 6 64 from site 1008:1011 arg1 unmodified 979:988 unmodified 979:988 unmodified 979:988 Protein C is unmodified at the equivalent site (Thr-68) and is completely beta-hydroxylated at Asp-71. 1544894 7 65 theme O-linked 1149:1156 arg1 fucose 1158:1163 O-linked fucose 1149:1163 O-linked fucose 1149:1163 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 66 link O-linked 1149:1156 arg1 fucose 1158:1163 O-linked fucose 1149:1163 O-linked fucose 1149:1163 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 3 67 theme post-translational 684:701 arg1 beta-hydroxylation 717:734 beta-hydroxylation 717:734 beta-hydroxylation of Asp-71 717:744 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 3 67 theme post-translational 684:701 arg1 modification 703:714 another post-translational modification 676:714 another post-translational modification 676:714 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 1544894 0 68 theme growth 50:55 arg1 factor 57:62 the first epidermal growth factor 30:62 the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains 30:140 O-linked fucose is present in the first epidermal growth factor domain of factor XII but not protein C. Epidermal growth factor (EGF) domains are found in many proteins, particularly those of the coagulation/fibrinolytic system. 1544894 7 69 contain has 1145:1147 arg1 VII 1141:1143 human factor VII 1128:1143 human factor VII 1128:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 69 contain has 1145:1147 arg2 fucose 1158:1163 O-linked fucose 1149:1163 O-linked fucose 1149:1163 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 7 69 contain has 1145:1147 arg1 domain 1118:1123 the first EGF domain 1104:1123 the first EGF domain of human factor VII 1104:1143 It has been recently reported that the first EGF domain of human factor VII has O-linked fucose at the equivalent position (Ser-60) (Bjoern, S., Foster, D. C., Thim, L., Wiberg, F. C., Christensen, M., Komiyama, Y., Pedersen, A. H., and Kisiel, W. (1991) J. Biol. 1544894 4 70 theme fucose 872:877 arg1 presence 851:858 the presence 847:858 the presence of O-linked fucose and/or beta-hydroxyaspartate 847:906 We isolated peptides containing these sites to determine, primarily by mass spectrometric analysis, the presence of O-linked fucose and/or beta-hydroxyaspartate. 1544894 3 71 theme modification 703:714 arg1 position 664:671 the position 660:671 the position of another post-translational modification, beta-hydroxylation of Asp-71 660:744 In protein C, this site is 3 residues from the position of another post-translational modification, beta-hydroxylation of Asp-71. 16497731 6 0 theme conserved 960:968 arg1 proline 970:976 multiple conserved proline and lysine residues 951:996 proline 970:976 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 4 1 theme multimerization 628:642 arg1 regulation 602:611 regulation 602:611 regulation of adiponectin multimerization and secretion 602:656 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 11 2 theme glucose-induced 1757:1771 arg1 increases 1773:1781 glucose-induced increases 1757:1781 glucose-induced increases in HMW multimer production from human adipose explants 1757:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 9 3 theme prolyl- 1488:1494 arg1 inhibitor 1475:1483 a pharmacological inhibitor 1457:1483 a pharmacological inhibitor of prolyl- and lysyl-hydroxylases 1457:1517 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 9 3 theme prolyl- 1488:1494 arg1 2,2'-dipyridyl 1520:1533 2,2'-dipyridyl 1520:1533 2,2'-dipyridyl 1520:1533 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 2 4 theme HMW 412:414 arg1 multimers 416:424 these HMW multimers 406:424 these HMW multimers 406:424 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 2 4 theme HMW 412:414 arg1 forms 458:462 the more bioactive forms 439:462 the more bioactive forms 439:462 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 9 5 theme pharmacological 1459:1473 arg1 inhibitor 1475:1483 a pharmacological inhibitor 1457:1483 a pharmacological inhibitor of prolyl- and lysyl-hydroxylases 1457:1517 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 9 5 theme pharmacological 1459:1473 arg1 2,2'-dipyridyl 1520:1533 2,2'-dipyridyl 1520:1533 2,2'-dipyridyl 1520:1533 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 9 6 theme lysyl-hydroxylases 1500:1517 arg1 inhibitor 1475:1483 a pharmacological inhibitor 1457:1483 a pharmacological inhibitor of prolyl- and lysyl-hydroxylases 1457:1517 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 9 6 theme lysyl-hydroxylases 1500:1517 arg1 2,2'-dipyridyl 1520:1533 2,2'-dipyridyl 1520:1533 2,2'-dipyridyl 1520:1533 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 6 7 theme bovine 907:912 arg1 adiponectin 914:924 murine and bovine adiponectin 896:924 murine and bovine adiponectin 896:924 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 8 8 theme recombinant 1291:1301 arg1 adiponectin 1309:1319 recombinant human adiponectin 1291:1319 recombinant human adiponectin expressed in mammalian cell lines 1291:1353 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 11 9 theme isoforms 1924:1931 arg1 profile 1901:1907 the two-dimensional electrophoresis profile 1865:1907 the two-dimensional electrophoresis profile of adiponectin isoforms 1865:1931 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 7 10 theme additional 1139:1148 arg1 hydroxylations 1150:1163 three additional hydroxylations 1133:1163 three additional hydroxylations on Pro71, Pro76, and Pro95 1133:1190 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 6 11 gly glycosylation 1002:1014 arg1 residues 989:996 multiple conserved proline and lysine residues 951:996 residues 989:996 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 6 11 gly glycosylation 1002:1014 arg1 hydroxylysines 1019:1032 hydroxylysines 1019:1032 hydroxylysines 1019:1032 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 6 11 gly glycosylation 1002:1014 arg1 proline 970:976 multiple conserved proline and lysine residues 951:996 proline 970:976 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 9 12 from Mutation 1356:1363 arg1 domain 1404:1409 the collagenous domain 1388:1409 the collagenous domain 1388:1409 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 6 13 theme hydroxylysines 1019:1032 arg1 hydroxylation 934:946 hydroxylation 934:946 hydroxylation of multiple conserved proline and lysine residues 934:996 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 6 13 theme hydroxylysines 1019:1032 arg1 glycosylation 1002:1014 glycosylation 1002:1014 glycosylation of hydroxylysines 1002:1032 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 8 14 theme adiponectin 1309:1319 arg1 secretion 1278:1286 secretion 1278:1286 secretion 1278:1286 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 8 14 theme adiponectin 1309:1319 arg1 formation 1264:1272 multimer formation 1255:1272 multimer formation 1255:1272 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 10 15 theme isoforms 1680:1687 arg1 lack 1648:1651 a complete lack 1637:1651 a complete lack of differentially modified isoforms 1637:1687 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 5 16 theme intertrimer 751:761 arg1 bonds 773:777 intertrimer disulfide bonds 751:777 intertrimer disulfide bonds 751:777 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 6 17 theme residues 989:996 arg1 hydroxylation 934:946 hydroxylation 934:946 hydroxylation of multiple conserved proline and lysine residues 934:996 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 6 17 theme residues 989:996 arg1 glycosylation 1002:1014 glycosylation 1002:1014 glycosylation of hydroxylysines 1002:1032 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 8 18 theme cell 1344:1347 arg1 lines 1349:1353 mammalian cell lines 1334:1353 mammalian cell lines 1334:1353 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 0 19 theme posttranslational 150:166 arg1 modifications 168:180 posttranslational modifications 150:180 posttranslational modifications 150:180 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 11 20 from changes 1854:1860 arg1 profile 1901:1907 the two-dimensional electrophoresis profile 1865:1907 the two-dimensional electrophoresis profile of adiponectin isoforms 1865:1931 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 9 21 theme multimers 1576:1584 arg1 formation 1546:1554 formation 1546:1554 formation of hexamers and HMW multimers 1546:1584 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 2 22 theme more 443:446 arg1 multimers 416:424 these HMW multimers 406:424 these HMW multimers 406:424 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 2 22 theme more 443:446 arg1 forms 458:462 the more bioactive forms 439:462 the more bioactive forms 439:462 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 11 23 from increases 1773:1781 arg1 production 1799:1808 HMW multimer production 1786:1808 HMW multimer production from human adipose explants 1786:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 11 23 from increases 1773:1781 arg1 explants 1829:1836 human adipose explants 1815:1836 human adipose explants 1815:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 11 24 theme electrophoresis 1885:1899 arg1 profile 1901:1907 the two-dimensional electrophoresis profile 1865:1907 the two-dimensional electrophoresis profile of adiponectin isoforms 1865:1931 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 5 25 theme multimers 816:824 arg1 formation 786:794 the formation 782:794 the formation of hexamers and HMW multimers 782:824 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 7 26 attach presence 1074:1081 arg1 adiponectin 1106:1116 human adiponectin 1100:1116 human adiponectin 1100:1116 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 7 26 attach presence 1074:1081 arg2 PTMs 1092:1095 these PTMs 1086:1095 these PTMs 1086:1095 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 12 27 theme physiological 2050:2062 arg1 factors 2064:2070 physiological factors 2050:2070 physiological factors 2050:2070 Collectively, these data suggest that adiponectin multimer composition is affected by changes in PTM in response to physiological factors. 16497731 12 28 from changes 2020:2026 arg1 PTM 2031:2033 PTM 2031:2033 PTM 2031:2033 Collectively, these data suggest that adiponectin multimer composition is affected by changes in PTM in response to physiological factors. 16497731 12 28 from changes 2020:2026 arg1 response 2038:2045 response 2038:2045 response to physiological factors 2038:2070 Collectively, these data suggest that adiponectin multimer composition is affected by changes in PTM in response to physiological factors. 16497731 0 29 from lysines 54:60 arg1 domain 81:86 the collagenous domain 65:86 the collagenous domain 65:86 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 9 30 theme modified 1368:1375 arg1 lysines 1377:1383 modified lysines 1368:1383 modified lysines 1368:1383 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 0 31 theme collagenous 69:79 arg1 domain 81:86 the collagenous domain 65:86 the collagenous domain 65:86 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 10 32 theme larger 1730:1735 arg1 multimers 1737:1745 larger multimers 1730:1745 larger multimers 1730:1745 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 0 33 theme Adiponectin 0:10 arg1 multimerization 12:26 Adiponectin multimerization 0:26 Adiponectin multimerization 0:26 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 3 34 theme adiponectin 489:499 arg1 composition 474:484 Multimer composition 465:484 Multimer composition of adiponectin 465:499 Multimer composition of adiponectin appears to be regulated; however, the molecular mechanisms involved are unknown. 16497731 5 35 theme PTMs 860:863 arg1 unknown 868:874 unknown 868:874 unknown 868:874 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 5 35 theme PTMs 860:863 arg1 role 846:849 the role 842:849 the role of other PTMs 842:863 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 9 36 theme collagenous 1392:1402 arg1 domain 1404:1409 the collagenous domain 1388:1409 the collagenous domain 1388:1409 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 1 37 dep secreted 200:207 arg1 multimeric 210:219 multimeric 210:219 multimeric 210:219 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 1 38 with protein 221:227 arg1 properties 293:302 antiinflammatory properties 276:302 antiinflammatory properties 276:302 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 1 38 with protein 221:227 arg1 antiatherogenic 255:269 antiatherogenic 255:269 antiatherogenic 255:269 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 1 38 with protein 221:227 arg1 insulin-sensitizing 234:252 insulin-sensitizing 234:252 insulin-sensitizing 234:252 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 0 39 dep lysines 54:60 arg1 evidence 89:96 evidence 89:96 conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications 44:180 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 10 40 theme human 1609:1613 arg1 adiponectin 1615:1625 Bacterially expressed human adiponectin 1587:1625 Bacterially expressed human adiponectin 1587:1625 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 11 41 theme HMW 1786:1788 arg1 multimer 1790:1797 HMW multimer 1786:1797 HMW multimer production from human adipose explants 1786:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 8 42 from role 1218:1221 arg1 secretion 1278:1286 secretion 1278:1286 secretion 1278:1286 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 8 42 from role 1218:1221 arg1 formation 1264:1272 multimer formation 1255:1272 multimer formation 1255:1272 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 9 43 theme multimers 1438:1446 arg1 formation 1421:1429 formation 1421:1429 formation of HMW multimers 1421:1446 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 12 44 theme adiponectin 1972:1982 arg1 multimer 1984:1991 adiponectin multimer 1972:1991 adiponectin multimer composition 1972:2003 Collectively, these data suggest that adiponectin multimer composition is affected by changes in PTM in response to physiological factors. 16497731 0 45 from alterations 135:145 arg1 modifications 168:180 posttranslational modifications 150:180 posttranslational modifications 150:180 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 10 46 theme complete 1639:1646 arg1 lack 1648:1651 a complete lack 1637:1651 a complete lack of differentially modified isoforms 1637:1687 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 11 47 theme human 1815:1819 arg1 explants 1829:1836 human adipose explants 1815:1836 human adipose explants 1815:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 6 48 theme proline 970:976 arg1 hydroxylation 934:946 hydroxylation 934:946 hydroxylation of multiple conserved proline and lysine residues 934:996 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 6 48 theme proline 970:976 arg1 glycosylation 1002:1014 glycosylation 1002:1014 glycosylation of hydroxylysines 1002:1032 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 10 49 dep bona 1708:1711 arg1 fide 1713:1716 fide 1713:1716 fide 1713:1716 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 1 50 theme antiinflammatory 276:291 arg1 properties 293:302 antiinflammatory properties 276:302 antiinflammatory properties 276:302 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 6 51 theme multiple 951:958 arg1 proline 970:976 multiple conserved proline and lysine residues 951:996 proline 970:976 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 10 52 theme modified 1671:1678 arg1 isoforms 1680:1687 differentially modified isoforms 1656:1687 differentially modified isoforms 1656:1687 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 4 53 theme adiponectin 616:626 arg1 multimerization 628:642 adiponectin multimerization 616:642 adiponectin multimerization 616:642 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 11 54 from explants 1829:1836 arg1 production 1799:1808 HMW multimer production 1786:1808 HMW multimer production from human adipose explants 1786:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 11 54 from explants 1829:1836 arg1 increases 1773:1781 glucose-induced increases 1757:1781 glucose-induced increases in HMW multimer production from human adipose explants 1757:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 1 55 theme secreted 200:207 arg1 protein 221:227 a secreted, multimeric protein 198:227 a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties 198:302 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 1 55 theme secreted 200:207 arg1 Adiponectin 183:193 Adiponectin 183:193 Adiponectin 183:193 Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. 16497731 8 56 theme human 1303:1307 arg1 adiponectin 1309:1319 recombinant human adiponectin 1291:1319 recombinant human adiponectin expressed in mammalian cell lines 1291:1353 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 11 57 theme adiponectin 1912:1922 arg1 isoforms 1924:1931 adiponectin isoforms 1912:1931 adiponectin isoforms 1912:1931 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 7 58 theme human 1100:1104 arg1 adiponectin 1106:1116 human adiponectin 1100:1116 human adiponectin 1100:1116 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 4 59 from changes 669:675 arg1 PTMs 713:716 PTMs 713:716 PTMs 713:716 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 4 59 from changes 669:675 arg1 modifications 698:710 posttranslational modifications 680:710 posttranslational modifications (PTMs) 680:717 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 8 60 theme lysines 1244:1250 arg1 role 1218:1221 the role 1214:1221 the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines 1214:1353 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 8 61 theme modified 1235:1242 arg1 lysines 1244:1250 the five modified lysines 1226:1250 the five modified lysines 1226:1250 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 4 62 theme posttranslational 680:696 arg1 PTMs 713:716 PTMs 713:716 PTMs 713:716 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 4 62 theme posttranslational 680:696 arg1 modifications 698:710 posttranslational modifications 680:710 posttranslational modifications (PTMs) 680:717 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 10 63 theme bona 1708:1711 arg1 trimers 1718:1724 bona fide trimers 1708:1724 bona fide trimers 1708:1724 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 7 64 theme PTMs 1092:1095 arg1 presence 1074:1081 the presence 1070:1081 the presence of these PTMs in human adiponectin 1070:1116 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 9 65 theme hexamers 1559:1566 arg1 formation 1546:1554 formation 1546:1554 formation of hexamers and HMW multimers 1546:1584 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 8 66 theme multimer 1255:1262 arg1 formation 1264:1272 multimer formation 1255:1272 multimer formation 1255:1272 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 6 67 theme lysine 982:987 arg1 residues 989:996 multiple conserved proline and lysine residues 951:996 residues 989:996 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 8 68 theme mammalian 1334:1342 arg1 lines 1349:1353 mammalian cell lines 1334:1353 mammalian cell lines 1334:1353 We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. 16497731 5 69 theme structural 731:740 arg1 role 742:745 a structural role 729:745 a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers 729:824 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 2 70 theme Serum 305:309 arg1 adiponectin 311:321 Serum adiponectin 305:321 Serum adiponectin 305:321 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 9 71 theme HMW 1572:1574 arg1 multimers 1576:1584 HMW multimers 1572:1584 HMW multimers 1572:1584 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 5 72 theme hexamers 799:806 arg1 formation 786:794 the formation 782:794 the formation of hexamers and HMW multimers 782:824 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 5 73 theme HMW 812:814 arg1 multimers 816:824 HMW multimers 812:824 HMW multimers 812:824 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 7 74 theme mass 1038:1041 arg1 spectrometry 1043:1054 mass spectrometry 1038:1054 mass spectrometry 1038:1054 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 0 75 theme multimerization 116:130 arg1 regulation 102:111 regulation 102:111 regulation of multimerization by alterations in posttranslational modifications 102:180 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 11 76 theme two-dimensional 1869:1883 arg1 profile 1901:1907 the two-dimensional electrophoresis profile 1865:1907 the two-dimensional electrophoresis profile of adiponectin isoforms 1865:1931 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 3 77 theme Multimer 465:472 arg1 composition 474:484 Multimer composition 465:484 Multimer composition of adiponectin 465:499 Multimer composition of adiponectin appears to be regulated; however, the molecular mechanisms involved are unknown. 16497731 9 78 theme lysines 1377:1383 arg1 Mutation 1356:1363 Mutation 1356:1363 Mutation of modified lysines in the collagenous domain 1356:1409 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 5 79 theme disulfide 763:771 arg1 bonds 773:777 intertrimer disulfide bonds 751:777 intertrimer disulfide bonds 751:777 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 5 80 theme other 854:858 arg1 PTMs 860:863 other PTMs 854:863 other PTMs 854:863 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 2 81 theme bioactive 448:456 arg1 multimers 416:424 these HMW multimers 406:424 these HMW multimers 406:424 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 2 81 theme bioactive 448:456 arg1 forms 458:462 the more bioactive forms 439:462 the more bioactive forms 439:462 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 7 82 from hydroxylations 1150:1163 arg1 Pro76 1175:1179 Pro76 1175:1179 Pro76 1175:1179 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 7 82 from hydroxylations 1150:1163 arg1 Pro95 1186:1190 Pro95 1186:1190 Pro95 1186:1190 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 7 82 from hydroxylations 1150:1163 arg1 Pro71 1168:1172 Pro71 1168:1172 Pro71 1168:1172 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 11 83 theme multimer 1790:1797 arg1 production 1799:1808 HMW multimer production 1786:1808 HMW multimer production from human adipose explants 1786:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 0 84 theme conserved 44:52 arg1 lysines 54:60 conserved lysines 44:60 conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications 44:180 Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. 16497731 6 85 theme murine 896:901 arg1 adiponectin 914:924 murine and bovine adiponectin 896:924 murine and bovine adiponectin 896:924 PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. 16497731 10 86 theme expressed 1599:1607 arg1 adiponectin 1615:1625 Bacterially expressed human adiponectin 1587:1625 Bacterially expressed human adiponectin 1587:1625 Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. 16497731 11 87 theme adipose 1821:1827 arg1 explants 1829:1836 human adipose explants 1815:1836 human adipose explants 1815:1836 Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. 16497731 9 88 theme HMW 1434:1436 arg1 multimers 1438:1446 HMW multimers 1434:1446 HMW multimers 1434:1446 Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. 16497731 12 89 theme multimer 1984:1991 arg1 composition 1993:2003 adiponectin multimer composition 1972:2003 adiponectin multimer composition 1972:2003 Collectively, these data suggest that adiponectin multimer composition is affected by changes in PTM in response to physiological factors. 16497731 4 90 theme secretion 648:656 arg1 regulation 602:611 regulation 602:611 regulation of adiponectin multimerization and secretion 602:656 We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). 16497731 2 91 theme larger 356:361 arg1 multimers 391:399 larger high-molecular-weight (HMW) multimers 356:399 larger high-molecular-weight (HMW) multimers 356:399 Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. 16497731 5 92 from role 742:745 arg1 formation 786:794 the formation 782:794 the formation of hexamers and HMW multimers 782:824 Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. 16497731 7 93 from presence 1074:1081 arg1 adiponectin 1106:1116 human adiponectin 1100:1116 human adiponectin 1100:1116 By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. 16497731 3 94 theme molecular 539:547 arg1 mechanisms 549:558 the molecular mechanisms 535:558 the molecular mechanisms involved 535:567 Multimer composition of adiponectin appears to be regulated; however, the molecular mechanisms involved are unknown. 16497731 3 94 theme molecular 539:547 arg1 unknown 573:579 unknown 573:579 unknown 573:579 Multimer composition of adiponectin appears to be regulated; however, the molecular mechanisms involved are unknown. 2963625 1 0 theme protein 138:144 arg1 sequence 87:94 The complete amino acid sequence 63:94 The complete amino acid sequence of the human complement system regulatory protein, factor H, 63:155 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 3 1 located found 429:433 arg1 number 446:451 a large number 438:451 a large number of other complement and non-complement proteins 438:499 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 3 1 located found 429:433 arg2 those 423:427 those 423:427 those 423:427 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 3 2 theme other 456:460 arg1 complement 462:471 other complement 456:471 other complement 456:471 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 5 3 theme homologous 687:696 arg1 unit 705:708 the seventh homologous repeat unit 675:708 the seventh homologous repeat unit of factor H 675:720 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 7 4 theme splicing 898:905 arg1 mechanism 907:915 an alternative splicing mechanism 883:915 an alternative splicing mechanism 883:915 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 4 5 theme outdated 604:611 arg1 plasma 613:618 outdated plasma 604:618 outdated plasma 604:618 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 4 6 theme cDNA 550:553 arg1 sequence 555:562 the cDNA sequence 546:562 the cDNA sequence 546:562 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 2 7 theme homologous 278:287 arg1 units 289:293 20 homologous units 275:293 20 homologous units 275:293 The sequence consists of 1213 amino acids arranged in 20 homologous units, each about 60 amino acids long, and an 18-residue leader sequence. 2963625 2 8 theme amino 251:255 arg1 acids 257:261 1213 amino acids 246:261 1213 amino acids arranged in 20 homologous units 246:293 The sequence consists of 1213 amino acids arranged in 20 homologous units, each about 60 amino acids long, and an 18-residue leader sequence. 2963625 3 9 theme non-complement 477:490 arg1 proteins 492:499 non-complement proteins 477:499 non-complement proteins 477:499 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 5 10 located observed 659:666 arg1 unit 705:708 the seventh homologous repeat unit 675:708 the seventh homologous repeat unit of factor H 675:720 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 5 10 located observed 659:666 arg2 polymorphism 642:653 A tyrosine/histidine polymorphism 621:653 A tyrosine/histidine polymorphism 621:653 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 1 11 theme factor 147:152 arg1 protein 138:144 the human complement system regulatory protein 99:144 the human complement system regulatory protein 99:144 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 11 theme factor 147:152 arg1 H 154:154 factor H 147:154 factor H 147:154 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 3 12 theme proteins 492:499 arg1 number 446:451 a large number 438:451 a large number of other complement and non-complement proteins 438:499 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 0 13 theme complete 4:11 arg1 sequence 24:31 The complete amino acid sequence 0:31 The complete amino acid sequence of human complement factor H 0:60 The complete amino acid sequence of human complement factor H. 2963625 5 14 theme factor 713:718 arg1 H 720:720 factor H 713:720 factor H 713:720 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 1 15 theme complete 67:74 arg1 sequence 87:94 The complete amino acid sequence 63:94 The complete amino acid sequence of the human complement system regulatory protein, factor H, 63:155 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 3 16 theme 60-amino-acid-long 367:384 arg1 units 397:401 The 60-amino-acid-long repetitive units 363:401 The 60-amino-acid-long repetitive units 363:401 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 3 16 theme 60-amino-acid-long 367:384 arg1 homologous 407:416 homologous 407:416 homologous 407:416 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 4 17 theme C-terminal 512:521 arg1 residues 523:530 Two basic C-terminal residues 502:530 Two basic C-terminal residues 502:530 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 4 18 attach isolated 590:597 arg1 plasma 613:618 outdated plasma 604:618 outdated plasma 604:618 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 4 18 attach isolated 590:597 arg2 H 588:588 factor H 581:588 factor H isolated from outdated plasma 581:618 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 7 19 theme H 979:979 arg1 mRNA 981:984 factor H mRNA 972:984 factor H mRNA 972:984 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 1 20 theme amino 76:80 arg1 sequence 87:94 The complete amino acid sequence 63:94 The complete amino acid sequence of the human complement system regulatory protein, factor H, 63:155 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 3 21 theme repetitive 386:395 arg1 units 397:401 The 60-amino-acid-long repetitive units 363:401 The 60-amino-acid-long repetitive units 363:401 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 3 21 theme repetitive 386:395 arg1 homologous 407:416 homologous 407:416 homologous 407:416 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 0 22 theme acid 19:22 arg1 sequence 24:31 The complete amino acid sequence 0:31 The complete amino acid sequence of human complement factor H 0:60 The complete amino acid sequence of human complement factor H. 2963625 4 23 theme basic 506:510 arg1 residues 523:530 Two basic C-terminal residues 502:530 Two basic C-terminal residues 502:530 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 3 24 with homologous 407:416 arg1 those 423:427 those 423:427 those 423:427 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 1 25 theme acid 82:85 arg1 sequence 87:94 The complete amino acid sequence 63:94 The complete amino acid sequence of the human complement system regulatory protein, factor H, 63:155 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 4 26 from H 588:588 arg1 absent 569:574 absent 569:574 absent 569:574 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 4 27 theme factor 581:586 arg1 H 588:588 factor H 581:588 factor H isolated from outdated plasma 581:618 Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. 2963625 0 28 theme amino 13:17 arg1 sequence 24:31 The complete amino acid sequence 0:31 The complete amino acid sequence of human complement factor H 0:60 The complete amino acid sequence of human complement factor H. 2963625 3 29 theme complement 462:471 arg1 number 446:451 a large number 438:451 a large number of other complement and non-complement proteins 438:499 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 7 30 theme factor 972:977 arg1 mRNA 981:984 factor H mRNA 972:984 factor H mRNA 972:984 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 6 31 theme H 813:813 arg1 variants 794:801 the two major allelic variants 772:801 the two major allelic variants of factor H 772:813 This is likely to represent a difference between the two major allelic variants of factor H. 2963625 2 32 theme 18-residue 335:344 arg1 sequence 353:360 an 18-residue leader sequence 332:360 an 18-residue leader sequence 332:360 The sequence consists of 1213 amino acids arranged in 20 homologous units, each about 60 amino acids long, and an 18-residue leader sequence. 2963625 2 33 theme amino 310:314 arg1 acids 316:320 each about 60 amino acids 296:320 each about 60 amino acids long 296:325 The sequence consists of 1213 amino acids arranged in 20 homologous units, each about 60 amino acids long, and an 18-residue leader sequence. 2963625 5 34 theme seventh 679:685 arg1 unit 705:708 the seventh homologous repeat unit 675:708 the seventh homologous repeat unit of factor H 675:720 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 6 35 theme factor 806:811 arg1 H 813:813 factor H 806:813 factor H 806:813 This is likely to represent a difference between the two major allelic variants of factor H. 2963625 2 36 theme leader 346:351 arg1 sequence 353:360 an 18-residue leader sequence 332:360 an 18-residue leader sequence 332:360 The sequence consists of 1213 amino acids arranged in 20 homologous units, each about 60 amino acids long, and an 18-residue leader sequence. 2963625 0 37 theme complement 42:51 arg1 H 60:60 human complement factor H 36:60 human complement factor H 36:60 The complete amino acid sequence of human complement factor H. 2963625 7 38 theme mRNA 981:984 arg1 species 961:967 at least two species 948:967 at least two species of factor H mRNA 948:984 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 7 39 theme species 961:967 arg1 formation 935:943 the formation 931:943 the formation of at least two species of factor H mRNA 931:984 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 5 40 theme H 720:720 arg1 unit 705:708 the seventh homologous repeat unit 675:708 the seventh homologous repeat unit of factor H 675:720 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 3 41 theme large 440:444 arg1 number 446:451 a large number 438:451 a large number of other complement and non-complement proteins 438:499 The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. 2963625 0 42 theme human 36:40 arg1 H 60:60 human complement factor H 36:60 human complement factor H 36:60 The complete amino acid sequence of human complement factor H. 2963625 5 43 theme tyrosine/histidine 623:640 arg1 polymorphism 642:653 A tyrosine/histidine polymorphism 621:653 A tyrosine/histidine polymorphism 621:653 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 2963625 1 44 attach derived 166:172 arg2 sequence 87:94 The complete amino acid sequence 63:94 The complete amino acid sequence of the human complement system regulatory protein, factor H, 63:155 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 44 attach derived 166:172 arg1 clones 213:218 sequencing three overlapping cDNA clones 179:218 sequencing three overlapping cDNA clones 179:218 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 7 45 theme alternative 886:896 arg1 mechanism 907:915 an alternative splicing mechanism 883:915 an alternative splicing mechanism 883:915 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 6 46 theme allelic 786:792 arg1 variants 794:801 the two major allelic variants 772:801 the two major allelic variants of factor H 772:813 This is likely to represent a difference between the two major allelic variants of factor H. 2963625 1 47 theme human 103:107 arg1 protein 138:144 the human complement system regulatory protein 99:144 the human complement system regulatory protein 99:144 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 47 theme human 103:107 arg1 H 154:154 factor H 147:154 factor H 147:154 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 6 48 theme major 780:784 arg1 variants 794:801 the two major allelic variants 772:801 the two major allelic variants of factor H 772:813 This is likely to represent a difference between the two major allelic variants of factor H. 2963625 1 49 theme complement 109:118 arg1 protein 138:144 the human complement system regulatory protein 99:144 the human complement system regulatory protein 99:144 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 49 theme complement 109:118 arg1 H 154:154 factor H 147:154 factor H 147:154 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 7 50 theme cDNA 834:837 arg1 clones 839:844 the cDNA clones 830:844 the cDNA clones 830:844 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 1 51 theme overlapping 196:206 arg1 clones 213:218 sequencing three overlapping cDNA clones 179:218 sequencing three overlapping cDNA clones 179:218 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 0 52 theme H 60:60 arg1 sequence 24:31 The complete amino acid sequence 0:31 The complete amino acid sequence of human complement factor H 0:60 The complete amino acid sequence of human complement factor H. 2963625 1 53 theme sequencing 179:188 arg1 clones 213:218 sequencing three overlapping cDNA clones 179:218 sequencing three overlapping cDNA clones 179:218 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 54 theme system 120:125 arg1 protein 138:144 the human complement system regulatory protein 99:144 the human complement system regulatory protein 99:144 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 54 theme system 120:125 arg1 H 154:154 factor H 147:154 factor H 147:154 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 7 55 theme clones 839:844 arg1 nature 820:825 The nature 816:825 The nature of the cDNA clones 816:844 The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA. 2963625 1 56 theme cDNA 208:211 arg1 clones 213:218 sequencing three overlapping cDNA clones 179:218 sequencing three overlapping cDNA clones 179:218 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 0 57 theme factor 53:58 arg1 H 60:60 human complement factor H 36:60 human complement factor H 36:60 The complete amino acid sequence of human complement factor H. 2963625 1 58 theme regulatory 127:136 arg1 protein 138:144 the human complement system regulatory protein 99:144 the human complement system regulatory protein 99:144 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 1 58 theme regulatory 127:136 arg1 H 154:154 factor H 147:154 factor H 147:154 The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. 2963625 5 59 theme repeat 698:703 arg1 unit 705:708 the seventh homologous repeat unit 675:708 the seventh homologous repeat unit of factor H 675:720 A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. 22344443 1 0 theme heart 176:180 arg1 rate 182:185 heart rate 176:185 heart rate 176:185 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 4 1 theme receptor 683:690 arg1 region 669:674 the transmembrane region 651:674 the transmembrane region of the receptor 651:690 Access is gained by ligands entering laterally between helices I and VII within the transmembrane region of the receptor. 22344443 2 2 theme crystal 309:315 arg1 structure 317:325 the crystal structure 305:325 the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic 305:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 3 3 theme receptor 561:568 arg1 terminus 521:528 amino terminus 515:528 amino terminus 515:528 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 3 3 theme receptor 561:568 arg1 loops 548:552 extracellular loops 534:552 extracellular loops 534:552 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 3 4 theme binding 481:487 arg1 pocket 489:494 the binding pocket 477:494 the binding pocket 477:494 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 1 5 theme lyso-phospholipid 61:77 arg1 1-phosphate 91:101 The lyso-phospholipid sphingosine 1-phosphate 57:101 The lyso-phospholipid sphingosine 1-phosphate 57:101 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 5 6 theme hydrophobic 867:877 arg1 volume 879:884 hydrophobic volume 867:884 hydrophobic volume 867:884 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 5 7 theme requirement 851:861 arg1 view 813:816 a detailed view 802:816 a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses 802:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 5 8 theme agonist 733:739 arg1 data 773:776 agonist structure-activity relationship data 733:776 agonist structure-activity relationship data 733:776 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 2 9 from T4-lysozyme 378:388 arg1 complex 406:412 complex 406:412 complex with an antagonist sphingolipid mimic 406:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 1 10 theme sphingosine 79:89 arg1 1-phosphate 91:101 The lyso-phospholipid sphingosine 1-phosphate 57:101 The lyso-phospholipid sphingosine 1-phosphate 57:101 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 2 11 with complex 406:412 arg1 sphingolipid 433:444 an antagonist sphingolipid mimic 419:450 an antagonist sphingolipid mimic 419:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 2 12 theme sphingosine 334:344 arg1 receptor 358:365 the sphingosine 1-phosphate receptor 1 330:367 the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic 330:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 1 13 theme vascular 192:199 arg1 tone 201:204 vascular tone 192:204 vascular tone 192:204 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 0 14 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a lipid G protein-coupled receptor 0:54 Crystal structure of a lipid G protein-coupled receptor. 22344443 2 15 dep sphingolipid 433:444 arg1 mimic 446:450 mimic 446:450 mimic 446:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 4 16 theme transmembrane 655:667 arg1 region 669:674 the transmembrane region 651:674 the transmembrane region of the receptor 651:690 Access is gained by ligands entering laterally between helices I and VII within the transmembrane region of the receptor. 22344443 2 17 theme antagonist 422:431 arg1 sphingolipid 433:444 an antagonist sphingolipid mimic 419:450 an antagonist sphingolipid mimic 419:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 5 18 theme detailed 804:811 arg1 view 813:816 a detailed view 802:816 a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses 802:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 1 19 theme lymphocyte 113:122 arg1 trafficking 124:134 lymphocyte trafficking 113:134 lymphocyte trafficking 113:134 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 5 20 theme immune 940:945 arg1 responses 964:972 immune and stromal cell responses 940:972 immune and stromal cell responses 940:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 5 21 theme stromal 951:957 arg1 responses 964:972 immune and stromal cell responses 940:972 immune and stromal cell responses 940:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 5 22 theme molecular 825:833 arg1 recognition 835:845 the molecular recognition 821:845 the molecular recognition 821:845 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 5 23 theme recognition 835:845 arg1 view 813:816 a detailed view 802:816 a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses 802:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 3 24 theme Extracellular 453:465 arg1 access 467:472 Extracellular access 453:472 Extracellular access to the binding pocket 453:494 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 0 25 theme receptor 47:54 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a lipid G protein-coupled receptor 0:54 Crystal structure of a lipid G protein-coupled receptor. 22344443 3 26 dep terminus 521:528 arg1 the 511:513 the 511:513 the 511:513 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 0 27 theme protein-coupled 31:45 arg1 receptor 47:54 a lipid G protein-coupled receptor 21:54 a lipid G protein-coupled receptor 21:54 Crystal structure of a lipid G protein-coupled receptor. 22344443 4 28 dep helices 626:632 arg1 helices 626:632 helices I and VII 626:642 helices I and VII within the transmembrane region of the receptor 626:690 Access is gained by ligands entering laterally between helices I and VII within the transmembrane region of the receptor. 22344443 4 28 dep helices 626:632 arg1 VII 640:642 VII 640:642 VII 640:642 Access is gained by ligands entering laterally between helices I and VII within the transmembrane region of the receptor. 22344443 4 28 dep helices 626:632 arg1 I 634:634 I 634:634 I 634:634 Access is gained by ligands entering laterally between helices I and VII within the transmembrane region of the receptor. 22344443 1 29 theme endothelial 137:147 arg1 development 149:159 endothelial development 137:159 endothelial development 137:159 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 5 30 theme responses 964:972 arg1 modulation 926:935 the modulation 922:935 the modulation of immune and stromal cell responses 922:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 2 31 theme receptor 358:365 arg1 structure 317:325 the crystal structure 305:325 the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic 305:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 5 32 theme cell 959:962 arg1 responses 964:972 immune and stromal cell responses 940:972 immune and stromal cell responses 940:972 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 3 33 theme extracellular 534:546 arg1 loops 548:552 extracellular loops 534:552 extracellular loops 534:552 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 2 34 theme 1-phosphate 346:356 arg1 receptor 358:365 the sphingosine 1-phosphate receptor 1 330:367 the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic 330:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 1 35 theme protein-coupled 237:251 arg1 receptors 277:285 G protein-coupled sphingosine 1-phosphate receptors 235:285 G protein-coupled sphingosine 1-phosphate receptors 235:285 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 5 36 theme structure-activity 741:758 arg1 data 773:776 agonist structure-activity relationship data 733:776 agonist structure-activity relationship data 733:776 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 2 37 theme S1P 391:393 arg1 T4-lysozyme 378:388 T4-lysozyme 378:388 T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic 378:450 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 2 37 theme S1P 391:393 arg1 -T4L 397:400 S1P(1)-T4L 391:400 S1P(1)-T4L 391:400 Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. 22344443 1 38 theme sphingosine 253:263 arg1 1-phosphate 265:275 sphingosine 1-phosphate 253:275 G protein-coupled sphingosine 1-phosphate receptors 235:285 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 22344443 5 39 theme relationship 760:771 arg1 data 773:776 agonist structure-activity relationship data 733:776 agonist structure-activity relationship data 733:776 This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses. 22344443 3 40 theme amino 515:519 arg1 terminus 521:528 amino terminus 515:528 amino terminus 515:528 Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. 22344443 1 41 theme 1-phosphate 265:275 arg1 receptors 277:285 G protein-coupled sphingosine 1-phosphate receptors 235:285 G protein-coupled sphingosine 1-phosphate receptors 235:285 The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. 15750791 4 0 theme Edg-1/S1P1 620:629 arg1 Edg-1/S1P1 620:629 Edg-1/S1P1 620:629 Edg-1/S1P1 620:629 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 4 0 theme Edg-1/S1P1 620:629 arg1 ectodomain 606:615 the N-terminal ectodomain 591:615 the N-terminal ectodomain of Edg-1/S1P1 591:629 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 0 1 theme 1-phosphate-stimulated 71:92 arg1 cells 94:98 sphingosine 1-phosphate-stimulated cells 59:98 sphingosine 1-phosphate-stimulated cells 59:98 Roles for N-glycosylation in the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells. 15750791 3 2 theme recent 370:375 arg1 studies 383:389 Our recent novel studies 366:389 Our recent novel studies 366:389 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 0 3 from Roles 0:4 arg1 dynamics 33:40 the dynamics 29:40 the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells 29:98 Roles for N-glycosylation in the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells. 15750791 3 4 gly glycosylated 422:433 arg1 portion 467:473 its N-terminal extracellular portion 438:473 its N-terminal extracellular portion 438:473 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 3 4 gly glycosylated 422:433 arg1 Edg-1/S1P1 408:417 Edg-1/S1P1 408:417 Edg-1/S1P1 408:417 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 3 4 gly glycosylated 422:433 arg2 Edg-1/S1P1 408:417 Edg-1/S1P1 408:417 Edg-1/S1P1 408:417 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 3 5 theme novel 377:381 arg1 studies 383:389 Our recent novel studies 366:389 Our recent novel studies 366:389 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 5 6 from role 756:759 arg1 dynamics 799:806 the dynamics 795:806 the dynamics 795:806 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 5 6 from role 756:759 arg1 movements 858:866 its lateral and internal movements 833:866 its lateral and internal movements within the membrane 833:886 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 4 7 theme cell 665:668 arg1 surface 670:676 the cell surface 661:676 the cell surface 661:676 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 5 8 theme ligand-stimulated 892:908 arg1 cells 920:924 ligand-stimulated mammalian cells 892:924 ligand-stimulated mammalian cells 892:924 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 1 9 attach released 165:172 arg1 platelets 189:197 activated platelets 179:197 activated platelets 179:197 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 9 attach released 165:172 arg2 mediator 156:163 a bioactive lipid mediator 138:163 a bioactive lipid mediator released from activated platelets 138:197 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 9 attach released 165:172 arg2 1-phosphate 113:123 Sphingosine 1-phosphate 101:123 Sphingosine 1-phosphate (Sph-1-P) 101:133 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 10 theme activated 179:187 arg1 platelets 189:197 activated platelets 179:197 activated platelets 179:197 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 3 11 theme N-terminal 442:451 arg1 portion 467:473 its N-terminal extracellular portion 438:473 its N-terminal extracellular portion 438:473 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 5 12 theme lateral 837:843 arg1 movements 858:866 its lateral and internal movements 833:866 its lateral and internal movements within the membrane 833:886 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 1 13 theme Sphingosine 101:111 arg1 mediator 156:163 a bioactive lipid mediator 138:163 a bioactive lipid mediator released from activated platelets 138:197 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 13 theme Sphingosine 101:111 arg1 Sph-1-P 126:132 Sph-1-P 126:132 Sph-1-P 126:132 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 13 theme Sphingosine 101:111 arg1 1-phosphate 113:123 Sphingosine 1-phosphate 101:123 Sphingosine 1-phosphate (Sph-1-P) 101:133 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 3 14 theme extracellular 453:465 arg1 portion 467:473 its N-terminal extracellular portion 438:473 its N-terminal extracellular portion 438:473 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 5 15 theme regulatory 745:754 arg1 role 756:759 a possible regulatory role 734:759 a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane 734:886 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 5 16 theme internal 849:856 arg1 movements 858:866 its lateral and internal movements 833:866 its lateral and internal movements within the membrane 833:886 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 4 17 theme ectodomain 606:615 arg1 structure 578:586 the structure 574:586 the structure of the N-terminal ectodomain of Edg-1/S1P1 574:629 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 3 18 gly glycosylation 511:523 arg2 site 525:528 the specific glycosylation site 498:528 the specific glycosylation site 498:528 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 3 19 theme specific 502:509 arg1 site 525:528 the specific glycosylation site 498:528 the specific glycosylation site 498:528 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 5 20 theme receptor 815:822 arg1 dynamics 799:806 the dynamics 795:806 the dynamics 795:806 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 5 20 theme receptor 815:822 arg1 movements 858:866 its lateral and internal movements 833:866 its lateral and internal movements within the membrane 833:886 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 4 21 theme N-terminal 595:604 arg1 Edg-1/S1P1 620:629 Edg-1/S1P1 620:629 Edg-1/S1P1 620:629 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 4 21 theme N-terminal 595:604 arg1 ectodomain 606:615 the N-terminal ectodomain 591:615 the N-terminal ectodomain of Edg-1/S1P1 591:629 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 5 22 theme mammalian 910:918 arg1 cells 920:924 ligand-stimulated mammalian cells 892:924 ligand-stimulated mammalian cells 892:924 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 5 23 from N-glycan 769:776 arg1 Edg-1/S1P1 781:790 Edg-1/S1P1 781:790 Edg-1/S1P1 781:790 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 0 24 theme Edg-1/S1P1 45:54 arg1 dynamics 33:40 the dynamics 29:40 the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells 29:98 Roles for N-glycosylation in the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells. 15750791 0 25 from dynamics 33:40 arg1 cells 94:98 sphingosine 1-phosphate-stimulated cells 59:98 sphingosine 1-phosphate-stimulated cells 59:98 Roles for N-glycosylation in the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 receptors 240:248 5 seven-transmembrane-spanning receptors 209:248 5 seven-transmembrane-spanning receptors 209:248 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 Edg-1/S1P1 251:260 Edg-1/S1P1 251:260 Edg-1/S1P1 251:260 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 Edg-6/S1P4 287:296 Edg-6/S1P4 287:296 Edg-6/S1P4 287:296 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 Edg-8/S1P5 302:311 Edg-8/S1P5 302:311 Edg-8/S1P5 302:311 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 Edg-3/S1P3 263:272 Edg-3/S1P3 263:272 Edg-3/S1P3 263:272 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 receptors 355:363 specific Sph-1-P receptors 338:363 specific Sph-1-P receptors 338:363 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 26 theme seven-transmembrane-spanning 211:238 arg1 Edg-5/S1P2 275:284 Edg-5/S1P2 275:284 Edg-5/S1P2 275:284 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 27 theme Sph-1-P 347:353 arg1 receptors 355:363 specific Sph-1-P receptors 338:363 specific Sph-1-P receptors 338:363 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 27 theme Sph-1-P 347:353 arg1 receptors 240:248 5 seven-transmembrane-spanning receptors 209:248 5 seven-transmembrane-spanning receptors 209:248 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 28 theme specific 338:345 arg1 receptors 355:363 specific Sph-1-P receptors 338:363 specific Sph-1-P receptors 338:363 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 2 28 theme specific 338:345 arg1 receptors 240:248 5 seven-transmembrane-spanning receptors 209:248 5 seven-transmembrane-spanning receptors 209:248 To date, 5 seven-transmembrane-spanning receptors, Edg-1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. 15750791 5 29 theme possible 736:743 arg1 role 756:759 a possible regulatory role 734:759 a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane 734:886 These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. 15750791 3 30 theme glycosylation 511:523 arg1 site 525:528 the specific glycosylation site 498:528 the specific glycosylation site 498:528 Our recent novel studies established that Edg-1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. 15750791 0 31 theme sphingosine 59:69 arg1 cells 94:98 sphingosine 1-phosphate-stimulated cells 59:98 sphingosine 1-phosphate-stimulated cells 59:98 Roles for N-glycosylation in the dynamics of Edg-1/S1P1 in sphingosine 1-phosphate-stimulated cells. 15750791 1 32 theme bioactive 140:148 arg1 1-phosphate 113:123 Sphingosine 1-phosphate 101:123 Sphingosine 1-phosphate (Sph-1-P) 101:133 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 32 theme bioactive 140:148 arg1 mediator 156:163 a bioactive lipid mediator 138:163 a bioactive lipid mediator released from activated platelets 138:197 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 4 33 theme N-glycosylation 686:700 arg1 process 702:708 the N-glycosylation process 682:708 the N-glycosylation process 682:708 We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. 15750791 1 34 theme lipid 150:154 arg1 1-phosphate 113:123 Sphingosine 1-phosphate 101:123 Sphingosine 1-phosphate (Sph-1-P) 101:133 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 15750791 1 34 theme lipid 150:154 arg1 mediator 156:163 a bioactive lipid mediator 138:163 a bioactive lipid mediator released from activated platelets 138:197 Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. 20044576 4 0 link O-linked 876:883 arg1 mannose 885:891 this phosphorylated O-linked mannose 856:891 this phosphorylated O-linked mannose 856:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 3 1 theme nuclear 460:466 arg1 resonance 477:485 mass spectrometry- and nuclear magnetic resonance 437:485 resonance 477:485 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 5 2 theme muscular 1075:1082 arg1 dystrophy 1084:1092 congenital muscular dystrophy 1064:1092 congenital muscular dystrophy 1064:1092 These findings expand our understanding of the mechanisms that underlie congenital muscular dystrophy. 20044576 3 3 theme mucin-like 576:585 arg1 domain 587:592 the mucin-like domain 572:592 the mucin-like domain of recombinant alpha-DG 572:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 3 3 theme mucin-like 576:585 arg1 alpha-DG 609:616 recombinant alpha-DG 597:616 recombinant alpha-DG 597:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 3 4 theme recombinant 597:607 arg1 alpha-DG 609:616 recombinant alpha-DG 597:616 recombinant alpha-DG 597:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 4 5 from defects 812:818 arg1 modification 840:851 a postphosphoryl modification 823:851 a postphosphoryl modification of this phosphorylated O-linked mannose 823:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 5 6 theme mechanisms 1039:1048 arg1 understanding 1018:1030 our understanding 1014:1030 our understanding of the mechanisms that underlie congenital muscular dystrophy 1014:1092 These findings expand our understanding of the mechanisms that underlie congenital muscular dystrophy. 20044576 3 7 theme structural 499:508 arg1 analyses 510:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 1 8 theme matrix 190:195 arg1 proteins 197:204 both extracellular matrix proteins 171:204 both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 171:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 3 9 theme resonance 477:485 arg1 analyses 510:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 1 10 theme extracellular 176:188 arg1 proteins 197:204 both extracellular matrix proteins 171:204 both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 171:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 4 11 theme muscle-eye-brain 694:709 arg1 disease 711:717 muscle-eye-brain disease 694:717 muscle-eye-brain disease 694:717 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 3 12 theme alpha-DG 609:616 arg1 domain 587:592 the mucin-like domain 572:592 the mucin-like domain of recombinant alpha-DG 572:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 3 12 theme alpha-DG 609:616 arg1 alpha-DG 609:616 recombinant alpha-DG 597:616 recombinant alpha-DG 597:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 2 13 theme muscular 411:418 arg1 dystrophy 420:428 congenital muscular dystrophy 400:428 congenital muscular dystrophy 400:428 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 0 14 theme O-mannosyl 0:9 arg1 phosphorylation 11:25 O-mannosyl phosphorylation 0:25 O-mannosyl phosphorylation of alpha-dystroglycan 0:47 O-mannosyl phosphorylation of alpha-dystroglycan is required for laminin binding. 20044576 2 15 theme congenital 400:409 arg1 dystrophy 420:428 congenital muscular dystrophy 400:428 congenital muscular dystrophy 400:428 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 0 16 theme alpha-dystroglycan 30:47 arg1 phosphorylation 11:25 O-mannosyl phosphorylation 0:25 O-mannosyl phosphorylation of alpha-dystroglycan 0:47 O-mannosyl phosphorylation of alpha-dystroglycan is required for laminin binding. 20044576 1 17 gly glycoprotein 130:141 arg1 Alpha-dystroglycan 82:99 Alpha-dystroglycan 82:99 Alpha-dystroglycan (alpha-DG) 82:110 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 1 17 gly glycoprotein 130:141 arg1 receptor 158:165 a receptor 156:165 a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 156:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 1 17 gly glycoprotein 130:141 arg1 glycoprotein 130:141 a cell-surface glycoprotein 115:141 a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 115:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 3 18 from glycan 562:567 arg1 domain 587:592 the mucin-like domain 572:592 the mucin-like domain of recombinant alpha-DG 572:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 3 18 from glycan 562:567 arg1 alpha-DG 609:616 recombinant alpha-DG 597:616 recombinant alpha-DG 597:616 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 4 19 theme O-linked 876:883 arg1 mannose 885:891 this phosphorylated O-linked mannose 856:891 this phosphorylated O-linked mannose 856:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 3 20 theme -based 492:497 arg1 analyses 510:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 2 21 with binding 355:361 arg1 laminin 368:374 laminin 368:374 laminin 368:374 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 2 22 theme posttranslational 309:325 arg1 modification 327:338 a posttranslational modification 307:338 a posttranslational modification 307:338 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 1 23 theme laminin-G 217:225 arg1 domains 227:233 laminin-G domains 217:233 laminin-G domains 217:233 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 3 24 theme spectrometry- 442:454 arg1 analyses 510:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses 437:517 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 4 25 theme Fukuyama 723:730 arg1 dystrophy 752:760 Fukuyama congenital muscular dystrophy 723:760 Fukuyama congenital muscular dystrophy 723:760 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 2 26 theme Receptor 261:268 arg1 binding 270:276 Receptor binding 261:276 Receptor binding 261:276 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 3 27 theme magnetic 468:475 arg1 resonance 477:485 mass spectrometry- and nuclear magnetic resonance 437:485 resonance 477:485 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 4 28 theme muscular 743:750 arg1 dystrophy 752:760 Fukuyama congenital muscular dystrophy 723:760 Fukuyama congenital muscular dystrophy 723:760 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 29 theme phosphorylated 861:874 arg1 mannose 885:891 this phosphorylated O-linked mannose 856:891 this phosphorylated O-linked mannose 856:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 0 30 theme laminin 65:71 arg1 binding 73:79 laminin binding 65:79 laminin binding 65:79 O-mannosyl phosphorylation of alpha-dystroglycan is required for laminin binding. 20044576 1 31 theme cell-surface 117:128 arg1 Alpha-dystroglycan 82:99 Alpha-dystroglycan 82:99 Alpha-dystroglycan (alpha-DG) 82:110 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 1 31 theme cell-surface 117:128 arg1 receptor 158:165 a receptor 156:165 a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 156:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 1 31 theme cell-surface 117:128 arg1 glycoprotein 130:141 a cell-surface glycoprotein 115:141 a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 115:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 4 32 contain have 807:810 arg1 patients 680:687 patients 680:687 patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy 680:760 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 32 contain have 807:810 arg2 defects 812:818 defects 812:818 defects in a postphosphoryl modification of this phosphorylated O-linked mannose 812:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 32 contain have 807:810 arg1 mice 774:777 mice 774:777 mice with myodystrophy 774:795 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 33 theme like-acetylglucosaminyltransferase 940:973 arg1 protein 983:989 the like-acetylglucosaminyltransferase (LARGE) protein 936:989 the like-acetylglucosaminyltransferase (LARGE) protein 936:989 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 34 theme mannose 885:891 arg1 modification 840:851 a postphosphoryl modification 823:851 a postphosphoryl modification of this phosphorylated O-linked mannose 823:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 3 35 theme phosphorylated 536:549 arg1 glycan 562:567 a phosphorylated O-mannosyl glycan 534:567 a phosphorylated O-mannosyl glycan 534:567 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 4 36 with patients 680:687 arg1 dystrophy 752:760 Fukuyama congenital muscular dystrophy 723:760 Fukuyama congenital muscular dystrophy 723:760 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 36 with patients 680:687 arg1 disease 711:717 muscle-eye-brain disease 694:717 muscle-eye-brain disease 694:717 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 36 with patients 680:687 arg1 myodystrophy 784:795 myodystrophy 784:795 myodystrophy 784:795 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 5 37 theme congenital 1064:1073 arg1 dystrophy 1084:1092 congenital muscular dystrophy 1064:1092 congenital muscular dystrophy 1064:1092 These findings expand our understanding of the mechanisms that underlie congenital muscular dystrophy. 20044576 1 38 theme certain 239:245 arg1 arenaviruses 247:258 certain arenaviruses 239:258 certain arenaviruses 239:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 4 39 theme LARGE 976:980 arg1 protein 983:989 the like-acetylglucosaminyltransferase (LARGE) protein 936:989 the like-acetylglucosaminyltransferase (LARGE) protein 936:989 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 40 theme postphosphoryl 825:838 arg1 modification 840:851 a postphosphoryl modification 823:851 a postphosphoryl modification of this phosphorylated O-linked mannose 823:891 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 2 41 theme defective 345:353 arg1 binding 355:361 defective binding 345:361 defective binding with laminin 345:374 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 1 42 contain containing 206:215 arg2 domains 227:233 laminin-G domains 217:233 laminin-G domains 217:233 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 1 42 contain containing 206:215 arg1 proteins 197:204 both extracellular matrix proteins 171:204 both extracellular matrix proteins containing laminin-G domains and certain arenaviruses 171:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 1 42 contain containing 206:215 arg2 arenaviruses 247:258 certain arenaviruses 239:258 certain arenaviruses 239:258 Alpha-dystroglycan (alpha-DG) is a cell-surface glycoprotein that acts as a receptor for both extracellular matrix proteins containing laminin-G domains and certain arenaviruses. 20044576 3 43 theme laminin 642:648 arg1 binding 650:656 laminin binding 642:656 laminin binding 642:656 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 3 44 theme O-mannosyl 551:560 arg1 glycan 562:567 a phosphorylated O-mannosyl glycan 534:567 a phosphorylated O-mannosyl glycan 534:567 Using mass spectrometry- and nuclear magnetic resonance (NMR)-based structural analyses, we identified a phosphorylated O-mannosyl glycan on the mucin-like domain of recombinant alpha-DG, which was required for laminin binding. 20044576 2 45 theme dystrophy 420:428 arg1 subclass 388:395 a subclass 386:395 a subclass of congenital muscular dystrophy 386:428 Receptor binding is thought to be mediated by a posttranslational modification, and defective binding with laminin underlies a subclass of congenital muscular dystrophy. 20044576 4 46 with mice 774:777 arg1 dystrophy 752:760 Fukuyama congenital muscular dystrophy 723:760 Fukuyama congenital muscular dystrophy 723:760 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 46 with mice 774:777 arg1 disease 711:717 muscle-eye-brain disease 694:717 muscle-eye-brain disease 694:717 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 46 with mice 774:777 arg1 myodystrophy 784:795 myodystrophy 784:795 myodystrophy 784:795 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 20044576 4 47 theme congenital 732:741 arg1 dystrophy 752:760 Fukuyama congenital muscular dystrophy 723:760 Fukuyama congenital muscular dystrophy 723:760 We demonstrated that patients with muscle-eye-brain disease and Fukuyama congenital muscular dystrophy, as well as mice with myodystrophy, commonly have defects in a postphosphoryl modification of this phosphorylated O-linked mannose, and that this modification is mediated by the like-acetylglucosaminyltransferase (LARGE) protein. 29106414 7 0 theme S1 1118:1119 arg1 conformations 1128:1140 S1 and S2 conformations 1118:1140 S1 and S2 conformations 1118:1140 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 0 1 theme distinct 70:77 arg1 states 79:84 three distinct states 64:84 three distinct states 64:84 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 0 2 from structures 8:17 arg1 states 79:84 three distinct states 64:84 three distinct states 64:84 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 7 3 theme luminal 1031:1037 arg1 pH 1039:1040 luminal pH 1031:1040 luminal pH 1031:1040 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 1 4 theme critical 152:159 arg1 role 161:164 a critical role 150:164 a critical role 150:164 TRPML3 channels are mainly localized to endolysosomes and play a critical role in the endocytic pathway. 29106414 3 5 theme cryo-EM 356:362 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 6 6 theme pore 721:724 arg1 loop 726:729 a luminal pore loop 711:729 a luminal pore loop 711:729 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 3 7 from structures 365:374 arg1 states 447:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 6 8 theme luminal 713:719 arg1 loop 726:729 a luminal pore loop 711:729 a luminal pore loop 711:729 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 2 9 from deafness 217:224 arg1 mice 254:257 mice 254:257 mice 254:257 Their dysfunction causes deafness and pigmentation defects in mice. 29106414 6 10 theme gating 890:895 arg1 knob 897:900 a 'gating knob' 887:901 a 'gating knob' 887:901 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 3 11 theme microscopy 344:353 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 6 12 theme pH. 799:801 arg1 S2 803:804 low pH. S2 795:804 low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob' 795:901 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 6 13 theme low 795:797 arg1 S2 803:804 low pH. S2 795:804 low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob' 795:901 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 3 14 theme closed 399:404 arg1 states 447:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 3 15 dep endolysosomal 296:308 arg1 present 322:328 present 322:328 present 322:328 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 0 16 theme Cryo-EM 0:6 arg1 structures 8:17 Cryo-EM structures 0:17 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states 0:84 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 3 17 theme low-pH-inhibited 430:445 arg1 states 447:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 3 18 theme human 379:383 arg1 TRPML3 385:390 human TRPML3 379:390 human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 379:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 3 19 from TRPML3 385:390 arg1 states 447:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 5 20 theme polycystin-mucolipin 581:600 arg1 PMD 610:612 PMD 610:612 PMD 610:612 A polycystin-mucolipin domain (PMD) forms a luminal cap. 29106414 5 20 theme polycystin-mucolipin 581:600 arg1 domain 602:607 A polycystin-mucolipin domain 579:607 A polycystin-mucolipin domain (PMD) 579:613 A polycystin-mucolipin domain (PMD) forms a luminal cap. 29106414 3 21 theme agonist-activated 407:423 arg1 states 447:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 the closed, agonist-activated, and low-pH-inhibited states 395:452 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 3 22 theme TRPML3 385:390 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 7 23 theme other 1046:1050 arg1 modulators 1066:1075 other physiological modulators 1046:1075 other physiological modulators such as PIP2 1046:1088 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 7 23 theme other 1046:1050 arg1 PIP2 1085:1088 PIP2 1085:1088 PIP2 1085:1088 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 1 24 theme TRPML3 87:92 arg1 channels 94:101 TRPML3 channels 87:101 TRPML3 channels 87:101 TRPML3 channels are mainly localized to endolysosomes and play a critical role in the endocytic pathway. 29106414 7 25 theme electrophysiological 967:986 arg1 studies 988:994 electrophysiological studies 967:994 electrophysiological studies 967:994 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 1 26 theme endocytic 173:181 arg1 pathway 183:189 the endocytic pathway 169:189 the endocytic pathway 169:189 TRPML3 channels are mainly localized to endolysosomes and play a critical role in the endocytic pathway. 29106414 4 27 theme agonist 517:523 arg1 ML-SA1 525:530 The agonist ML-SA1 513:530 The agonist ML-SA1 513:530 The agonist ML-SA1 lodges between S5 and S6 and opens an S6 gate. 29106414 3 28 with states 447:452 arg1 resolutions 460:470 resolutions 460:470 resolutions of 4.06, 3.62, and 4.65 Å 460:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 2 29 from defects 243:249 arg1 mice 254:257 mice 254:257 mice 254:257 Their dysfunction causes deafness and pigmentation defects in mice. 29106414 7 30 theme studies 988:994 arg1 results 956:962 the results 952:962 the results of electrophysiological studies 952:994 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 0 31 theme endolysosomal 32:44 arg1 channel 53:59 the human endolysosomal TRPML3 channel 22:59 the human endolysosomal TRPML3 channel 22:59 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 3 32 theme TRPML3 260:265 arg1 activity 267:274 TRPML3 activity 260:274 TRPML3 activity 260:274 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 2 33 theme pigmentation 230:241 arg1 defects 243:249 pigmentation defects 230:249 pigmentation defects 230:249 Their dysfunction causes deafness and pigmentation defects in mice. 29106414 0 34 theme human 26:30 arg1 channel 53:59 the human endolysosomal TRPML3 channel 22:59 the human endolysosomal TRPML3 channel 22:59 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 7 35 theme physiological 1052:1064 arg1 modulators 1066:1075 other physiological modulators 1046:1075 other physiological modulators such as PIP2 1046:1088 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 7 35 theme physiological 1052:1064 arg1 PIP2 1085:1088 PIP2 1085:1088 PIP2 1085:1088 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 3 36 from states 447:452 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 0 37 theme channel 53:59 arg1 structures 8:17 Cryo-EM structures 0:17 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states 0:84 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 6 38 theme intracellular 857:869 arg1 regions 871:877 several intracellular regions 849:877 several intracellular regions 849:877 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 3 39 theme endolysosomal 296:308 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 6 40 theme conformational 757:770 arg1 changes 772:778 dramatic conformational changes 748:778 dramatic conformational changes 748:778 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 0 41 theme TRPML3 46:51 arg1 channel 53:59 the human endolysosomal TRPML3 channel 22:59 the human endolysosomal TRPML3 channel 22:59 Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states. 29106414 6 42 theme several 849:855 arg1 regions 871:877 several intracellular regions 849:877 several intracellular regions 849:877 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 5 43 theme luminal 623:629 arg1 cap 631:633 a luminal cap 621:633 a luminal cap 621:633 A polycystin-mucolipin domain (PMD) forms a luminal cap. 29106414 6 44 theme dramatic 748:755 arg1 changes 772:778 dramatic conformational changes 748:778 dramatic conformational changes 748:778 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 7 45 theme new 1008:1010 arg1 mechanism 1012:1020 a new mechanism 1006:1020 a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations 1006:1140 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 6 46 theme gating 673:678 arg1 rod 680:682 a 'gating rod' 670:683 a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob' 670:901 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 7 47 theme unique 910:915 arg1 features 928:935 These unique structural features 904:935 These unique structural features 904:935 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 3 48 theme low 292:294 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 3 49 dep present 322:328 arg1 pH. 310:312 pH. 310:312 pH. 310:312 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 7 50 theme structural 917:926 arg1 features 928:935 These unique structural features 904:935 These unique structural features 904:935 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 7 51 theme S2 1125:1126 arg1 conformations 1128:1140 S1 and S2 conformations 1118:1140 S1 and S2 conformations 1118:1140 These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP2 regulate TRPML3 by changing S1 and S2 conformations. 29106414 6 52 dep S2 803:804 arg1 response 783:790 response 783:790 response 783:790 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 6 52 dep S2 803:804 arg1 extends 806:812 extends 806:812 extends intracellularly 806:828 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 6 52 dep S2 803:804 arg1 interacts 834:842 interacts 834:842 interacts with several intracellular regions to form a 'gating knob' 834:901 S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. 29106414 3 53 theme cryo-electron 330:342 arg1 structures 365:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures 292:374 low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å 292:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 3 54 theme Å 496:496 arg1 resolutions 460:470 resolutions 460:470 resolutions of 4.06, 3.62, and 4.65 Å 460:496 TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. 29106414 4 55 theme S6 570:571 arg1 gate 573:576 an S6 gate 567:576 an S6 gate 567:576 The agonist ML-SA1 lodges between S5 and S6 and opens an S6 gate. 3264725 9 0 theme factor 1851:1856 arg1 VIIa 1858:1861 human plasma factor VIIa 1838:1861 human plasma factor VIIa 1838:1861 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 1 from identical 1797:1805 arg1 VIIa 1858:1861 human plasma factor VIIa 1838:1861 human plasma factor VIIa 1838:1861 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 1 from identical 1797:1805 arg1 VIIa 1829:1832 recombinant factor VIIa 1810:1832 recombinant factor VIIa 1810:1832 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 2 from VIIa 1858:1861 arg1 compositions 1772:1783 the overall carbohydrate compositions 1747:1783 the overall carbohydrate compositions 1747:1783 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 2 from VIIa 1858:1861 arg1 identical 1797:1805 identical 1797:1805 identical 1797:1805 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 8 3 theme asparagine 1605:1614 arg1 residue 1616:1622 asparagine residue 145 1605:1626 asparagine residue 145 1605:1626 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 3 4 theme factor 720:725 arg1 VIIa 727:730 recombinant factor VIIa 708:730 recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line 708:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 6 5 theme acid 1224:1227 arg1 residues 1229:1236 the 10 N-terminally located glutamic acid residues 1187:1236 the 10 N-terminally located glutamic acid residues 1187:1236 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 5 theme acid 1224:1227 arg1 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 9 6 theme carbohydrate 1759:1770 arg1 compositions 1772:1783 the overall carbohydrate compositions 1747:1783 the overall carbohydrate compositions 1747:1783 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 6 theme carbohydrate 1759:1770 arg1 identical 1797:1805 identical 1797:1805 identical 1797:1805 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 1 7 gly glycoprotein 186:197 arg1 VII 157:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 7 gly glycoprotein 186:197 arg1 glycoprotein 186:197 a vitamin K dependent glycoprotein 164:197 a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor 164:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 6 8 theme VIIa 1405:1408 arg1 molecule 1410:1417 the recombinant factor VIIa molecule 1382:1417 the recombinant factor VIIa molecule 1382:1417 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 7 9 theme Asparagine 1420:1429 arg1 residues 1431:1438 Asparagine residues 145 and 322 1420:1450 residues 1431:1438 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 2 10 theme asparagine 602:611 arg1 residues 613:620 2 N-glycosylated asparagine residues 585:620 2 N-glycosylated asparagine residues 585:620 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 10 theme asparagine 602:611 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 9 11 theme minor 1689:1693 arg1 differences 1695:1705 minor differences 1689:1705 minor differences in the sialic acid and fucose contents 1689:1744 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 1 12 theme dependent 176:184 arg1 VII 157:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 12 theme dependent 176:184 arg1 glycoprotein 186:197 a vitamin K dependent glycoprotein 164:197 a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor 164:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 2 13 theme acid 523:526 arg1 residues 528:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 13 theme acid 523:526 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 4 14 theme acid 885:888 arg1 analysis 890:897 amino acid analysis 879:897 amino acid analysis 879:897 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 6 15 located found 1342:1346 arg2 residues 1328:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 15 located found 1342:1346 arg1 positions 1369:1377 the corresponding positions 1351:1377 the corresponding positions of the recombinant factor VIIa molecule 1351:1417 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 0 16 theme kidney 118:123 arg1 cells 125:129 transfected baby hamster kidney cells 93:129 transfected baby hamster kidney cells 93:129 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 1 17 theme tissue 349:354 arg1 factor 356:361 tissue factor 349:361 tissue factor 349:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 5 18 contain contain 1123:1129 arg2 acid 1152:1155 beta-hydroxyaspartic acid 1131:1155 beta-hydroxyaspartic acid 1131:1155 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 5 18 contain contain 1123:1129 arg1 VIIa 1105:1108 human plasma factor VIIa 1085:1108 human plasma factor VIIa 1085:1108 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 5 18 contain contain 1123:1129 arg1 VIIa 1076:1079 recombinant factor VIIa 1057:1079 recombinant factor VIIa 1057:1079 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 7 19 gly N-glycosylated 1475:1488 arg1 residues 1431:1438 Asparagine residues 145 and 322 1420:1450 residues 1431:1438 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 7 19 gly N-glycosylated 1475:1488 arg1 322 1448:1450 322 1448:1450 322 1448:1450 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 7 19 gly N-glycosylated 1475:1488 arg1 VIIa 1513:1516 human plasma factor VIIa 1493:1516 human plasma factor VIIa 1493:1516 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 7 19 gly N-glycosylated 1475:1488 arg2 322 1448:1450 322 1448:1450 322 1448:1450 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 7 19 gly N-glycosylated 1475:1488 arg2 residues 1431:1438 Asparagine residues 145 and 322 1420:1450 residues 1431:1438 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 2 20 theme acid 567:570 arg1 residue 572:578 1 beta-hydroxylated aspartic acid residue 538:578 1 beta-hydroxylated aspartic acid residue 538:578 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 20 theme acid 567:570 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 3 21 theme culture 753:759 arg1 medium 761:766 the culture medium 749:766 the culture medium of a transfected baby hamster kidney cell line 749:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 3 22 theme human 837:841 arg1 VIIa 857:860 human plasma factor VIIa 837:860 human plasma factor VIIa 837:860 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 2 23 theme beta-hydroxylated 540:556 arg1 residue 572:578 1 beta-hydroxylated aspartic acid residue 538:578 1 beta-hydroxylated aspartic acid residue 538:578 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 23 theme beta-hydroxylated 540:556 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 6 24 theme plasma 1167:1172 arg1 VIIa 1181:1184 human plasma factor VIIa 1161:1184 human plasma factor VIIa 1161:1184 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 3 25 theme hamster 790:796 arg1 line 810:813 a transfected baby hamster kidney cell line 771:813 a transfected baby hamster kidney cell line 771:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 1 26 theme Blood 132:136 arg1 VII 157:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 26 theme Blood 132:136 arg1 glycoprotein 186:197 a vitamin K dependent glycoprotein 164:197 a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor 164:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 9 27 theme overall 1751:1757 arg1 compositions 1772:1783 the overall carbohydrate compositions 1747:1783 the overall carbohydrate compositions 1747:1783 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 27 theme overall 1751:1757 arg1 identical 1797:1805 identical 1797:1805 identical 1797:1805 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 4 28 theme Edman 931:935 arg1 degradations 937:948 automated Edman degradations 921:948 automated Edman degradations 921:948 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 3 29 theme transfected 773:783 arg1 line 810:813 a transfected baby hamster kidney cell line 771:813 a transfected baby hamster kidney cell line 771:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 0 30 theme factor 65:70 arg1 VIIa 72:75 human factor VIIa 59:75 human factor VIIa 59:75 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 1 31 theme factor 311:316 arg1 IX 318:319 factor IX 311:319 factor IX 311:319 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 4 32 theme VIIa 994:997 arg1 backbone 963:970 the protein backbone 951:970 the protein backbone of recombinant factor VIIa 951:997 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 4 32 theme VIIa 994:997 arg1 identical 1015:1023 identical 1015:1023 identical 1015:1023 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 1 33 theme factor 150:155 arg1 VII 157:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 33 theme factor 150:155 arg1 glycoprotein 186:197 a vitamin K dependent glycoprotein 164:197 a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor 164:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 5 34 theme human 1085:1089 arg1 VIIa 1105:1108 human plasma factor VIIa 1085:1108 human plasma factor VIIa 1085:1108 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 3 35 theme factor 850:855 arg1 VIIa 857:860 human plasma factor VIIa 837:860 human plasma factor VIIa 837:860 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 4 36 theme mapping 908:914 arg1 use 866:868 use 866:868 use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations 866:948 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 3 37 theme cell 805:808 arg1 line 810:813 a transfected baby hamster kidney cell line 771:813 a transfected baby hamster kidney cell line 771:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 6 38 theme gamma-carboxylated 1309:1326 arg1 residues 1328:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 0 39 theme baby 105:108 arg1 kidney 118:123 baby hamster kidney 105:123 transfected baby hamster kidney cells 93:129 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 1 40 theme Ca2+ 340:343 arg1 presence 328:335 the presence 324:335 the presence of Ca2+ and tissue factor 324:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 4 41 theme factor 987:992 arg1 VIIa 994:997 recombinant factor VIIa 975:997 recombinant factor VIIa 975:997 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 8 42 dep glycosylated 1667:1678 arg1 % 1664:1664 approximately 66% 1648:1664 approximately 66% 1648:1664 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 0 43 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 6 44 theme full 1290:1293 arg1 residues 1328:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 2 45 theme posttranslational 389:405 arg1 modifications 407:419 potential posttranslational modifications 379:419 potential posttranslational modifications 379:419 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 6 46 theme recombinant 1386:1396 arg1 VIIa 1405:1408 the recombinant factor VIIa 1382:1408 the recombinant factor VIIa molecule 1382:1417 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 5 47 theme beta-hydroxyaspartic 1131:1150 arg1 acid 1152:1155 beta-hydroxyaspartic acid 1131:1155 beta-hydroxyaspartic acid 1131:1155 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 2 48 dep gamma-carboxylated 473:490 arg1 located 506:512 located 506:512 located 506:512 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 7 49 theme plasma 1499:1504 arg1 VIIa 1513:1516 human plasma factor VIIa 1493:1516 human plasma factor VIIa 1493:1516 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 4 50 theme protein 955:961 arg1 backbone 963:970 the protein backbone 951:970 the protein backbone of recombinant factor VIIa 951:997 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 4 50 theme protein 955:961 arg1 identical 1015:1023 identical 1015:1023 identical 1015:1023 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 8 51 gly glycosylated 1667:1678 arg1 residue 1616:1622 asparagine residue 145 1605:1626 asparagine residue 145 1605:1626 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 0 52 theme posttranslational 24:40 arg1 modifications 42:54 posttranslational modifications 24:54 posttranslational modifications 24:54 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 5 53 theme factor 1098:1103 arg1 VIIa 1105:1108 human plasma factor VIIa 1085:1108 human plasma factor VIIa 1085:1108 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 2 54 theme factor 440:445 arg1 residues 528:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 54 theme factor 440:445 arg1 residue 572:578 1 beta-hydroxylated aspartic acid residue 538:578 1 beta-hydroxylated aspartic acid residue 538:578 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 54 theme factor 440:445 arg1 residues 613:620 2 N-glycosylated asparagine residues 585:620 2 N-glycosylated asparagine residues 585:620 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 54 theme factor 440:445 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 8 55 theme recombinant 1526:1536 arg1 VIIa 1545:1548 the recombinant factor VIIa 1522:1548 the recombinant factor VIIa 1522:1548 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 9 56 theme sialic 1714:1719 arg1 acid 1721:1724 sialic acid 1714:1724 sialic acid 1714:1724 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 6 57 theme corresponding 1355:1367 arg1 positions 1369:1377 the corresponding positions 1351:1377 the corresponding positions of the recombinant factor VIIa molecule 1351:1417 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 58 theme located 1207:1213 arg1 residues 1229:1236 the 10 N-terminally located glutamic acid residues 1187:1236 the 10 N-terminally located glutamic acid residues 1187:1236 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 58 theme located 1207:1213 arg1 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 4 59 theme human 1030:1034 arg1 VIIa 1043:1046 human factor VIIa 1030:1046 human factor VIIa 1030:1046 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 3 60 theme acid 655:658 arg1 sequence 660:667 the amino acid sequence 645:667 the amino acid sequence 645:667 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 1 61 theme activated 212:220 arg1 form 222:225 its activated form 208:225 its activated form 208:225 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 61 theme activated 212:220 arg1 VIIa 235:238 factor VIIa 228:238 factor VIIa 228:238 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 8 62 gly glycosylated 1584:1595 arg1 VIIa 1545:1548 the recombinant factor VIIa 1522:1548 the recombinant factor VIIa 1522:1548 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 8 62 gly glycosylated 1584:1595 arg1 residue 1562:1568 asparagine residue 322 1551:1572 asparagine residue 322 1551:1572 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 8 62 gly glycosylated 1584:1595 arg2 residue 1562:1568 asparagine residue 322 1551:1572 asparagine residue 322 1551:1572 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 8 63 theme asparagine 1551:1560 arg1 residue 1562:1568 asparagine residue 322 1551:1572 asparagine residue 322 1551:1572 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 9 64 theme plasma 1844:1849 arg1 VIIa 1858:1861 human plasma factor VIIa 1838:1861 human plasma factor VIIa 1838:1861 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 7 65 from N-glycosylated 1475:1488 arg1 VIIa 1513:1516 human plasma factor VIIa 1493:1516 human plasma factor VIIa 1493:1516 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 3 66 theme VIIa 727:730 arg1 sequence 660:667 the amino acid sequence 645:667 the amino acid sequence 645:667 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 3 66 theme VIIa 727:730 arg1 modifications 691:703 posttranslational modifications 673:703 posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line 673:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 6 67 theme glutamic 1215:1222 arg1 residues 1229:1236 the 10 N-terminally located glutamic acid residues 1187:1236 the 10 N-terminally located glutamic acid residues 1187:1236 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 67 theme glutamic 1215:1222 arg1 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 2 68 theme human 434:438 arg1 residues 528:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 68 theme human 434:438 arg1 residue 572:578 1 beta-hydroxylated aspartic acid residue 538:578 1 beta-hydroxylated aspartic acid residue 538:578 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 68 theme human 434:438 arg1 residues 613:620 2 N-glycosylated asparagine residues 585:620 2 N-glycosylated asparagine residues 585:620 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 68 theme human 434:438 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 3 69 theme recombinant 708:718 arg1 VIIa 727:730 recombinant factor VIIa 708:730 recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line 708:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 2 70 gly N-glycosylated 587:600 arg1 residues 613:620 2 N-glycosylated asparagine residues 585:620 2 N-glycosylated asparagine residues 585:620 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 70 gly N-glycosylated 587:600 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 71 theme N-glycosylated 587:600 arg1 residues 613:620 2 N-glycosylated asparagine residues 585:620 2 N-glycosylated asparagine residues 585:620 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 71 theme N-glycosylated 587:600 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 9 72 dep acid 1721:1724 arg1 the 1710:1712 the 1710:1712 the 1710:1712 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 4 73 theme amino 879:883 arg1 analysis 890:897 amino acid analysis 879:897 amino acid analysis 879:897 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 0 74 from plasma 82:87 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 0 74 from plasma 82:87 arg1 modifications 42:54 posttranslational modifications 24:54 posttranslational modifications 24:54 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 9 75 from VIIa 1829:1832 arg1 compositions 1772:1783 the overall carbohydrate compositions 1747:1783 the overall carbohydrate compositions 1747:1783 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 75 from VIIa 1829:1832 arg1 identical 1797:1805 identical 1797:1805 identical 1797:1805 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 1 76 theme factor 356:361 arg1 presence 328:335 the presence 324:335 the presence of Ca2+ and tissue factor 324:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 9 77 theme factor 1822:1827 arg1 VIIa 1829:1832 recombinant factor VIIa 1810:1832 recombinant factor VIIa 1810:1832 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 4 78 theme analysis 890:897 arg1 use 866:868 use 866:868 use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations 866:948 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 9 79 theme fucose 1730:1735 arg1 contents 1737:1744 fucose contents 1730:1744 fucose contents 1730:1744 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 2 80 theme aspartic 558:565 arg1 residue 572:578 1 beta-hydroxylated aspartic acid residue 538:578 1 beta-hydroxylated aspartic acid residue 538:578 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 80 theme aspartic 558:565 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 6 81 theme factor 1174:1179 arg1 VIIa 1181:1184 human plasma factor VIIa 1161:1184 human plasma factor VIIa 1161:1184 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 3 82 theme plasma 843:848 arg1 VIIa 857:860 human plasma factor VIIa 837:860 human plasma factor VIIa 837:860 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 9 83 from differences 1695:1705 arg1 acid 1721:1724 sialic acid 1714:1724 sialic acid 1714:1724 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 9 83 from differences 1695:1705 arg1 contents 1737:1744 fucose contents 1730:1744 fucose contents 1730:1744 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 4 84 theme HPLC 873:876 arg1 use 866:868 use 866:868 use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations 866:948 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 6 85 theme human 1161:1165 arg1 VIIa 1181:1184 human plasma factor VIIa 1161:1184 human plasma factor VIIa 1161:1184 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 3 86 theme kidney 798:803 arg1 line 810:813 a transfected baby hamster kidney cell line 771:813 a transfected baby hamster kidney cell line 771:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 0 87 theme VIIa 72:75 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 0 87 theme VIIa 72:75 arg1 modifications 42:54 posttranslational modifications 24:54 posttranslational modifications 24:54 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 1 88 theme coagulation 138:148 arg1 VII 157:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII 132:159 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 88 theme coagulation 138:148 arg1 glycoprotein 186:197 a vitamin K dependent glycoprotein 164:197 a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor 164:361 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 4 89 theme automated 921:929 arg1 degradations 937:948 automated Edman degradations 921:948 automated Edman degradations 921:948 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 3 90 theme baby 785:788 arg1 line 810:813 a transfected baby hamster kidney cell line 771:813 a transfected baby hamster kidney cell line 771:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 4 91 theme degradations 937:948 arg1 use 866:868 use 866:868 use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations 866:948 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 0 92 theme transfected 93:103 arg1 cells 125:129 transfected baby hamster kidney cells 93:129 transfected baby hamster kidney cells 93:129 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 5 93 theme plasma 1091:1096 arg1 VIIa 1105:1108 human plasma factor VIIa 1085:1108 human plasma factor VIIa 1085:1108 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 4 94 theme peptide 900:906 arg1 mapping 908:914 peptide mapping 900:914 peptide mapping 900:914 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 3 95 theme line 810:813 arg1 medium 761:766 the culture medium 749:766 the culture medium of a transfected baby hamster kidney cell line 749:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 0 96 theme hamster 110:116 arg1 kidney 118:123 baby hamster kidney 105:123 transfected baby hamster kidney cells 93:129 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 4 97 with identical 1015:1023 arg1 VIIa 1043:1046 human factor VIIa 1030:1046 human factor VIIa 1030:1046 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 2 98 theme gamma-carboxylated 473:490 arg1 residues 528:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 98 theme gamma-carboxylated 473:490 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 6 99 theme partial 1301:1307 arg1 residues 1328:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 9 full and 1 partial gamma-carboxylated residues 1288:1335 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 0 100 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 5 101 theme factor 1069:1074 arg1 VIIa 1076:1079 recombinant factor VIIa 1057:1079 recombinant factor VIIa 1057:1079 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 4 102 theme recombinant 975:985 arg1 VIIa 994:997 recombinant factor VIIa 975:997 recombinant factor VIIa 975:997 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 6 103 theme factor 1398:1403 arg1 VIIa 1405:1408 the recombinant factor VIIa 1382:1408 the recombinant factor VIIa molecule 1382:1417 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 1 104 theme coagulation 261:271 arg1 process 273:279 the coagulation process 257:279 the coagulation process 257:279 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 7 105 from VIIa 1513:1516 arg1 N-glycosylated 1475:1488 N-glycosylated 1475:1488 N-glycosylated 1475:1488 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 6 106 located found 1243:1247 arg1 VIIa 1181:1184 human plasma factor VIIa 1161:1184 human plasma factor VIIa 1161:1184 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 106 located found 1243:1247 arg2 residues 1229:1236 the 10 N-terminally located glutamic acid residues 1187:1236 the 10 N-terminally located glutamic acid residues 1187:1236 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 6 106 located found 1243:1247 arg2 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 gamma-carboxylated 1261:1278 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 9 107 theme recombinant 1810:1820 arg1 VIIa 1829:1832 recombinant factor VIIa 1810:1832 recombinant factor VIIa 1810:1832 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 2 108 theme modifications 407:419 arg1 types 370:374 Three types 364:374 Three types of potential posttranslational modifications 364:419 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 3 109 theme present 630:636 arg1 study 638:642 the present study 626:642 the present study 626:642 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 2 110 theme potential 379:387 arg1 modifications 407:419 potential posttranslational modifications 379:419 potential posttranslational modifications 379:419 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 7 111 theme factor 1506:1511 arg1 VIIa 1513:1516 human plasma factor VIIa 1493:1516 human plasma factor VIIa 1493:1516 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 9 112 theme human 1838:1842 arg1 VIIa 1858:1861 human plasma factor VIIa 1838:1861 human plasma factor VIIa 1838:1861 Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa. 3264725 2 113 theme VIIa 447:450 arg1 residues 528:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 113 theme VIIa 447:450 arg1 residue 572:578 1 beta-hydroxylated aspartic acid residue 538:578 1 beta-hydroxylated aspartic acid residue 538:578 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 113 theme VIIa 447:450 arg1 residues 613:620 2 N-glycosylated asparagine residues 585:620 2 N-glycosylated asparagine residues 585:620 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 113 theme VIIa 447:450 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 8 114 theme factor 1538:1543 arg1 VIIa 1545:1548 the recombinant factor VIIa 1522:1548 the recombinant factor VIIa 1522:1548 In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. 3264725 0 115 theme human 59:63 arg1 VIIa 72:75 human factor VIIa 59:75 human factor VIIa 59:75 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 1 116 theme factor 295:300 arg1 X 302:302 factor X 295:302 factor X 295:302 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 7 117 theme human 1493:1497 arg1 VIIa 1513:1516 human plasma factor VIIa 1493:1516 human plasma factor VIIa 1493:1516 Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. 3264725 5 118 theme recombinant 1057:1067 arg1 VIIa 1076:1079 recombinant factor VIIa 1057:1079 recombinant factor VIIa 1057:1079 Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. 3264725 2 119 theme glutamic 514:521 arg1 residues 528:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 10 gamma-carboxylated, N-terminally located glutamic acid residues 470:535 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 2 119 theme glutamic 514:521 arg1 molecule 452:459 the human factor VIIa molecule 430:459 the human factor VIIa molecule 430:459 Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. 3264725 3 120 theme amino 649:653 arg1 sequence 660:667 the amino acid sequence 645:667 the amino acid sequence 645:667 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 4 121 theme factor 1036:1041 arg1 VIIa 1043:1046 human factor VIIa 1030:1046 human factor VIIa 1030:1046 By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. 3264725 0 122 from cells 125:129 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence 0:18 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 0 122 from cells 125:129 arg1 modifications 42:54 posttranslational modifications 24:54 posttranslational modifications 24:54 Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. 3264725 3 123 theme posttranslational 673:689 arg1 modifications 691:703 posttranslational modifications 673:703 posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line 673:813 In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. 3264725 6 124 theme molecule 1410:1417 arg1 positions 1369:1377 the corresponding positions 1351:1377 the corresponding positions of the recombinant factor VIIa molecule 1351:1417 In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. 3264725 1 125 theme factor 228:233 arg1 form 222:225 its activated form 208:225 its activated form 208:225 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 3264725 1 125 theme factor 228:233 arg1 VIIa 235:238 factor VIIa 228:238 factor VIIa 228:238 Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. 15606899 1 0 theme two-hybrid 142:151 arg1 system 153:158 a yeast two-hybrid system 134:158 a yeast two-hybrid system 134:158 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 4 1 dep convertases 676:686 arg1 furin 688:692 furin 688:692 the pro-protein convertases furin 660:692 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 4 1 dep convertases 676:686 arg1 PC5A 726:729 PC5A 726:729 PC5A 726:729 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 4 1 dep convertases 676:686 arg1 convertases 676:686 the pro-protein convertases 660:686 the pro-protein convertases furin 660:692 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 4 1 dep convertases 676:686 arg1 PC7 718:720 PC7 718:720 PC7 718:720 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 4 2 theme lesser 704:709 arg1 extent 711:716 a lesser extent 702:716 a lesser extent 702:716 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 0 3 theme binding 91:97 arg1 partners 99:106 binding partners 91:106 binding partners 91:106 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 0 4 dep processed 108:116 arg1 protein 77:83 brain-specific type II membrane protein BRI3 45:88 brain-specific type II membrane protein BRI3 45:88 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 0 4 dep processed 108:116 arg1 enzyme 32:37 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 1 5 theme C-terminal 243:252 arg1 tail 264:267 the C-terminal cytosolic tail 239:267 the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1 239:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 4 6 theme C-terminal 752:761 arg1 product 792:798 a C-terminal secreted approximately 4-kDa product 750:798 a C-terminal secreted approximately 4-kDa product 750:798 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 1 7 theme cytosolic 254:262 arg1 tail 264:267 the C-terminal cytosolic tail 239:267 the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1 239:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 2 8 theme type 412:415 arg1 BRI3 437:440 the brain-specific type II membrane protein BRI3 393:440 the brain-specific type II membrane protein BRI3 393:440 This identified seven potential candidates, including the brain-specific type II membrane protein BRI3. 15606899 4 9 theme 4-kDa 786:790 arg1 product 792:798 a C-terminal secreted approximately 4-kDa product 750:798 a C-terminal secreted approximately 4-kDa product 750:798 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 5 10 theme interacting 864:874 arg1 protein 876:882 its novel interacting protein pro-BRI3 854:891 its novel interacting protein pro-BRI3 854:891 Thus, furin efficiently processes both pro-BACE1 and its novel interacting protein pro-BRI3. 15606899 3 11 theme co-immunoprecipitation 463:484 arg1 experiments 486:496 Co-localization and co-immunoprecipitation experiments 443:496 experiments 486:496 Co-localization and co-immunoprecipitation experiments confirmed that BACE1 and BRI3 co-localize and interact with each other via the cytosolic tail of BACE1. 15606899 4 12 theme pulse 615:619 arg1 analyses 637:644 pulse and pulse-chase analyses 615:644 analyses 637:644 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 0 13 theme protein 13:19 arg1 enzyme 32:37 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 4 14 theme pro-protein 664:674 arg1 PC5A 726:729 PC5A 726:729 PC5A 726:729 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 4 14 theme pro-protein 664:674 arg1 PC7 718:720 PC7 718:720 PC7 718:720 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 4 14 theme pro-protein 664:674 arg1 convertases 676:686 the pro-protein convertases 660:686 the pro-protein convertases furin 660:692 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 0 15 theme Beta-amyloid 0:11 arg1 enzyme 32:37 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 2 16 theme potential 361:369 arg1 BRI3 437:440 the brain-specific type II membrane protein BRI3 393:440 the brain-specific type II membrane protein BRI3 393:440 This identified seven potential candidates, including the brain-specific type II membrane protein BRI3. 15606899 2 16 theme potential 361:369 arg1 candidates 371:380 seven potential candidates 355:380 seven potential candidates 355:380 This identified seven potential candidates, including the brain-specific type II membrane protein BRI3. 15606899 2 17 theme protein 429:435 arg1 BRI3 437:440 the brain-specific type II membrane protein BRI3 393:440 the brain-specific type II membrane protein BRI3 393:440 This identified seven potential candidates, including the brain-specific type II membrane protein BRI3. 15606899 1 18 theme human 175:179 arg1 library 192:198 a human brain cDNA library 173:198 a human brain cDNA library 173:198 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 0 19 theme converting 21:30 arg1 enzyme 32:37 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 0:39 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 3 20 theme cytosolic 577:585 arg1 tail 587:590 the cytosolic tail 573:590 the cytosolic tail of BACE1 573:599 Co-localization and co-immunoprecipitation experiments confirmed that BACE1 and BRI3 co-localize and interact with each other via the cytosolic tail of BACE1. 15606899 1 21 theme brain 181:185 arg1 library 192:198 a human brain cDNA library 173:198 a human brain cDNA library 173:198 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 22 theme beta-secretase 276:289 arg1 BACE 331:334 BACE 331:334 BACE 331:334 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 22 theme beta-secretase 276:289 arg1 enzyme 323:328 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 2 23 theme brain-specific 397:410 arg1 BRI3 437:440 the brain-specific type II membrane protein BRI3 393:440 the brain-specific type II membrane protein BRI3 393:440 This identified seven potential candidates, including the brain-specific type II membrane protein BRI3. 15606899 1 24 theme cDNA 187:190 arg1 library 192:198 a human brain cDNA library 173:198 a human brain cDNA library 173:198 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 25 theme beta-amyloid 291:302 arg1 BACE 331:334 BACE 331:334 BACE 331:334 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 25 theme beta-amyloid 291:302 arg1 enzyme 323:328 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 5 26 theme novel 858:862 arg1 protein 876:882 its novel interacting protein pro-BRI3 854:891 its novel interacting protein pro-BRI3 854:891 Thus, furin efficiently processes both pro-BACE1 and its novel interacting protein pro-BRI3. 15606899 0 27 theme type 60:63 arg1 protein 77:83 brain-specific type II membrane protein BRI3 45:88 brain-specific type II membrane protein BRI3 45:88 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 1 28 theme protein 304:310 arg1 BACE 331:334 BACE 331:334 BACE 331:334 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 28 theme protein 304:310 arg1 enzyme 323:328 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 3 29 theme BACE1 595:599 arg1 tail 587:590 the cytosolic tail 573:590 the cytosolic tail of BACE1 573:599 Co-localization and co-immunoprecipitation experiments confirmed that BACE1 and BRI3 co-localize and interact with each other via the cytosolic tail of BACE1. 15606899 0 30 theme brain-specific 45:58 arg1 protein 77:83 brain-specific type II membrane protein BRI3 45:88 brain-specific type II membrane protein BRI3 45:88 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 1 31 theme converting 312:321 arg1 BACE 331:334 BACE 331:334 BACE 331:334 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 31 theme converting 312:321 arg1 enzyme 323:328 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 the beta-secretase beta-amyloid protein converting enzyme (BACE)1 272:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 32 theme enzyme 323:328 arg1 tail 264:267 the C-terminal cytosolic tail 239:267 the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1 239:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 4 33 theme secreted 763:770 arg1 product 792:798 a C-terminal secreted approximately 4-kDa product 750:798 a C-terminal secreted approximately 4-kDa product 750:798 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 15606899 1 34 theme possible 204:211 arg1 proteins 225:232 possible interacting proteins 204:232 possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1 204:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 0 35 theme membrane 68:75 arg1 protein 77:83 brain-specific type II membrane protein BRI3 45:88 brain-specific type II membrane protein BRI3 45:88 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 3 36 theme Co-localization 443:457 arg1 experiments 486:496 Co-localization and co-immunoprecipitation experiments 443:496 experiments 486:496 Co-localization and co-immunoprecipitation experiments confirmed that BACE1 and BRI3 co-localize and interact with each other via the cytosolic tail of BACE1. 15606899 1 37 with proteins 225:232 arg1 tail 264:267 the C-terminal cytosolic tail 239:267 the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1 239:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 1 38 theme interacting 213:223 arg1 proteins 225:232 possible interacting proteins 204:232 possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1 204:336 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 0 39 theme II 65:66 arg1 protein 77:83 brain-specific type II membrane protein BRI3 45:88 brain-specific type II membrane protein BRI3 45:88 Beta-amyloid protein converting enzyme 1 and brain-specific type II membrane protein BRI3: binding partners processed by furin. 15606899 2 40 theme membrane 420:427 arg1 BRI3 437:440 the brain-specific type II membrane protein BRI3 393:440 the brain-specific type II membrane protein BRI3 393:440 This identified seven potential candidates, including the brain-specific type II membrane protein BRI3. 15606899 1 41 theme yeast 136:140 arg1 system 153:158 a yeast two-hybrid system 134:158 a yeast two-hybrid system 134:158 Using a yeast two-hybrid system, we screened a human brain cDNA library for possible interacting proteins with the C-terminal cytosolic tail of the beta-secretase beta-amyloid protein converting enzyme (BACE)1. 15606899 4 42 theme pulse-chase 625:635 arg1 analyses 637:644 pulse and pulse-chase analyses 615:644 analyses 637:644 Furthermore, pulse and pulse-chase analyses revealed that the pro-protein convertases furin, and to a lesser extent PC7 and PC5A, process BRI3 into a C-terminal secreted approximately 4-kDa product. 29581294 7 0 theme pronounced 1126:1135 arg1 rearrangements 1152:1165 pronounced conformational rearrangements 1126:1165 pronounced conformational rearrangements 1126:1165 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 4 1 theme lysyl 639:643 arg1 structure 620:628 the crystal structure 608:628 the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution 608:687 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 7 2 from residues 982:989 arg1 structure 998:1006 the structure 994:1006 the structure 994:1006 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 2 3 theme elastin 425:431 arg1 cross-linking 408:420 the cross-linking 404:420 the cross-linking of elastin and collagen 404:444 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 0 4 from structure 8:16 arg1 state 72:76 a precursor state 60:76 a precursor state 60:76 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 4 5 theme human 633:637 arg1 lysyl 639:643 human lysyl oxidase-like 2 633:658 human lysyl oxidase-like 2 (hLOXL2) 633:667 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 4 5 theme human 633:637 arg1 hLOXL2 661:666 hLOXL2 661:666 hLOXL2 661:666 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 1 6 theme oxidative 190:198 arg1 deamination 200:210 the oxidative deamination 186:210 the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens 186:299 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 8 7 theme structure-function 1296:1313 arg1 relationship 1315:1326 the structure-function relationship 1292:1326 the structure-function relationship of LOX proteins 1292:1342 The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. 29581294 3 8 theme physiological 498:510 arg1 significance 512:523 their physiological significance 492:523 their physiological significance 492:523 Despite their physiological significance, the structure of this important family of enzymes remains elusive. 29581294 0 9 theme precursor 62:70 arg1 state 72:76 a precursor state 60:76 a precursor state 60:76 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 5 10 from activity 805:812 arg1 assay 840:844 our in vitro assay 827:844 our in vitro assay 827:844 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 5 11 theme hLOXL2 817:822 arg1 activity 805:812 the enzymatic activity 791:812 the enzymatic activity of hLOXL2 in our in vitro assay 791:844 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 5 11 theme hLOXL2 817:822 arg1 generation 776:785 LTQ generation 772:785 LTQ generation 772:785 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 7 12 theme precursor 1101:1109 arg1 state 1111:1115 a precursor state 1099:1115 a precursor state 1099:1115 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 4 13 theme crystal 612:618 arg1 structure 620:628 the crystal structure 608:628 the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution 608:687 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 1 14 theme copper- 112:118 arg1 oxidase 168:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 5 15 theme copper-binding 708:721 arg1 hLOXL2 731:736 hLOXL2 731:736 hLOXL2 731:736 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 5 15 theme copper-binding 708:721 arg1 site 723:726 the copper-binding site 704:726 the copper-binding site of hLOXL2 704:736 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 4 16 from resolution 678:687 arg1 structure 620:628 the crystal structure 608:628 the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution 608:687 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 2 17 theme foundational 382:393 arg1 step 395:398 the foundational step 378:398 the foundational step for the cross-linking of elastin and collagen 378:444 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 7 18 theme conformational 1137:1150 arg1 rearrangements 1152:1165 pronounced conformational rearrangements 1126:1165 pronounced conformational rearrangements 1126:1165 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 8 19 theme proteins 1335:1342 arg1 relationship 1315:1326 the structure-function relationship 1292:1326 the structure-function relationship of LOX proteins 1292:1342 The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. 29581294 1 20 theme lysyl 124:128 arg1 oxidase 168:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 0 21 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 0:77 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 1 22 theme lysine 215:220 arg1 residues 222:229 lysine residues 215:229 lysine residues 215:229 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 8 23 theme LOX 1331:1333 arg1 proteins 1335:1342 LOX proteins 1331:1342 LOX proteins 1331:1342 The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. 29581294 5 24 theme enzymatic 795:803 arg1 activity 805:812 the enzymatic activity 791:812 the enzymatic activity of hLOXL2 in our in vitro assay 791:844 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 0 25 theme human 21:25 arg1 hLOXL2 49:54 hLOXL2 49:54 hLOXL2 49:54 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 0 25 theme human 21:25 arg1 lysyl 27:31 human lysyl oxidase-like 2 21:46 human lysyl oxidase-like 2 (hLOXL2) 21:55 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 1 26 theme residues 222:229 arg1 deamination 200:210 the oxidative deamination 186:210 the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens 186:299 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 2 27 theme lysine 360:365 arg1 deamination 345:355 The oxidative deamination 331:355 The oxidative deamination of lysine 331:365 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 2 27 theme lysine 360:365 arg1 crucial 458:464 crucial 458:464 crucial 458:464 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 6 28 theme copper 882:887 arg1 loading 889:895 copper loading 882:895 copper loading 882:895 Biochemical analysis confirms that copper loading robustly activates hLOXL2 and supports LTQ formation. 29581294 1 29 theme aldehyde 314:321 arg1 groups 323:328 aldehyde groups 314:328 aldehyde groups 314:328 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 30 theme Lysyl 79:83 arg1 LOXs 95:98 LOXs 95:98 LOXs 95:98 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 30 theme Lysyl 79:83 arg1 oxidases 85:92 Lysyl oxidases 79:92 Lysyl oxidases (LOXs) 79:99 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 30 theme Lysyl 79:83 arg1 type 104:107 a type 102:107 a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 102:174 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 7 31 theme present 1067:1073 arg1 structure 1075:1083 the present structure 1063:1083 the present structure 1063:1083 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 0 32 theme oxidase-like 33:44 arg1 hLOXL2 49:54 hLOXL2 49:54 hLOXL2 49:54 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 0 32 theme oxidase-like 33:44 arg1 lysyl 27:31 human lysyl oxidase-like 2 21:46 human lysyl oxidase-like 2 (hLOXL2) 21:55 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 1 33 theme extracellular 234:246 arg1 ECM 256:258 ECM 256:258 ECM 256:258 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 33 theme extracellular 234:246 arg1 matrix 248:253 extracellular matrix 234:253 extracellular matrix (ECM) proteins such as elastins and collagens 234:299 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 8 34 theme LOX-targeting 1364:1376 arg1 discovery 1383:1391 LOX-targeting drug discovery 1364:1391 LOX-targeting drug discovery 1364:1391 The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. 29581294 0 35 theme lysyl 27:31 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 0:77 Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state. 29581294 1 36 theme matrix 248:253 arg1 collagens 291:299 collagens 291:299 collagens 291:299 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 36 theme matrix 248:253 arg1 proteins 261:268 extracellular matrix (ECM) proteins 234:268 extracellular matrix (ECM) proteins such as elastins and collagens 234:299 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 36 theme matrix 248:253 arg1 elastins 278:285 elastins 278:285 elastins 278:285 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 2 37 theme ECM 470:472 arg1 modeling 474:481 ECM modeling 470:481 ECM modeling 470:481 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 5 38 theme in 831:832 arg1 assay 840:844 our in vitro assay 827:844 our in vitro assay 827:844 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 5 39 theme LTQ 772:774 arg1 generation 776:785 LTQ generation 772:785 LTQ generation 772:785 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 1 40 theme -dependent 151:160 arg1 oxidase 168:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 7 41 theme LTQ 968:970 arg1 residues 982:989 the LTQ precursor residues 964:989 the LTQ precursor residues in the structure 964:1006 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 5 42 theme hLOXL2 731:736 arg1 hLOXL2 731:736 hLOXL2 731:736 hLOXL2 731:736 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 5 42 theme hLOXL2 731:736 arg1 site 723:726 the copper-binding site 704:726 the copper-binding site of hLOXL2 704:736 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 6 43 theme Biochemical 847:857 arg1 analysis 859:866 Biochemical analysis 847:866 Biochemical analysis 847:866 Biochemical analysis confirms that copper loading robustly activates hLOXL2 and supports LTQ formation. 29581294 7 44 theme precursor 972:980 arg1 residues 982:989 the LTQ precursor residues 964:989 the LTQ precursor residues in the structure 964:1006 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 8 45 theme drug 1378:1381 arg1 discovery 1383:1391 LOX-targeting drug discovery 1364:1391 LOX-targeting drug discovery 1364:1391 The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. 29581294 4 46 theme oxidase-like 645:656 arg1 lysyl 639:643 human lysyl oxidase-like 2 633:658 human lysyl oxidase-like 2 (hLOXL2) 633:667 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 4 46 theme oxidase-like 645:656 arg1 hLOXL2 661:666 hLOXL2 661:666 hLOXL2 661:666 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 5 47 used occupied 741:748 arg2 hLOXL2 731:736 hLOXL2 731:736 hLOXL2 731:736 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 5 47 used occupied 741:748 arg2 site 723:726 the copper-binding site 704:726 the copper-binding site of hLOXL2 704:736 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 1 48 theme amine 162:166 arg1 oxidase 168:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 112:174 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 5 49 dep in 831:832 arg1 vitro 834:838 vitro 834:838 vitro 834:838 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 3 50 theme enzymes 568:574 arg1 family 558:563 this important family 543:563 this important family of enzymes 543:574 Despite their physiological significance, the structure of this important family of enzymes remains elusive. 29581294 4 51 theme 2.4-Å 672:676 arg1 resolution 678:687 2.4-Å resolution 672:687 2.4-Å resolution 672:687 Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. 29581294 8 52 theme important 1253:1261 arg1 foundation 1263:1272 an important foundation 1250:1272 an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery 1250:1391 The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. 29581294 1 53 theme oxidase 168:174 arg1 oxidases 85:92 Lysyl oxidases 79:92 Lysyl oxidases (LOXs) 79:99 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 1 53 theme oxidase 168:174 arg1 type 104:107 a type 102:107 a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase 102:174 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 2 54 theme oxidative 335:343 arg1 deamination 345:355 The oxidative deamination 331:355 The oxidative deamination of lysine 331:365 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 2 54 theme oxidative 335:343 arg1 crucial 458:464 crucial 458:464 crucial 458:464 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 3 55 theme important 548:556 arg1 family 558:563 this important family 543:563 this important family of enzymes 543:574 Despite their physiological significance, the structure of this important family of enzymes remains elusive. 29581294 2 56 theme collagen 437:444 arg1 cross-linking 408:420 the cross-linking 404:420 the cross-linking of elastin and collagen 404:444 The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. 29581294 1 57 theme proteins 261:268 arg1 deamination 200:210 the oxidative deamination 186:210 the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens 186:299 Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. 29581294 3 58 theme family 558:563 arg1 structure 530:538 the structure 526:538 the structure of this important family of enzymes 526:574 Despite their physiological significance, the structure of this important family of enzymes remains elusive. 29581294 5 59 from generation 776:785 arg1 assay 840:844 our in vitro assay 827:844 our in vitro assay 827:844 Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. 29581294 7 60 theme protein 1189:1195 arg1 activation 1197:1206 protein activation 1189:1206 protein activation 1189:1206 Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. 29581294 6 61 theme LTQ 936:938 arg1 formation 940:948 LTQ formation 936:948 LTQ formation 936:948 Biochemical analysis confirms that copper loading robustly activates hLOXL2 and supports LTQ formation. 16186819 1 0 contain have 130:133 arg1 Thrombospondins 74:88 Thrombospondins 74:88 Thrombospondins (THBSs) 74:96 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 1 0 contain have 130:133 arg1 glycoproteins 111:123 secreted glycoproteins 102:123 secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix 102:202 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 1 0 contain have 130:133 arg2 roles 139:143 key roles 135:143 key roles 135:143 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 2 1 theme factor-like 353:363 arg1 modules 365:371 three epidermal growth factor-like modules 330:371 three epidermal growth factor-like modules 330:371 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 1 theme factor-like 353:363 arg1 repeats 392:398 13 aspartate-rich repeats 374:398 13 aspartate-rich repeats 374:398 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 1 theme factor-like 353:363 arg1 module 418:423 a lectin-like module 404:423 a lectin-like module 404:423 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 2 theme human 302:306 arg1 THBS2 308:312 human THBS2 302:312 human THBS2 302:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 5 3 theme THBSs 707:711 arg1 alterations 692:702 genetic alterations 684:702 genetic alterations of THBSs 684:711 The structure suggests how genetic alterations of THBSs result in disease. 16186819 2 4 theme domain 292:297 arg1 structure 252:260 the 2.6-A-resolution crystal structure 223:260 the 2.6-A-resolution crystal structure 223:260 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 5 gly glycosylated 269:280 arg1 domain 292:297 the glycosylated signature domain 265:297 the glycosylated signature domain of human THBS2 265:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 5 gly glycosylated 269:280 arg1 THBS2 308:312 human THBS2 302:312 human THBS2 302:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 6 theme signature 282:290 arg1 domain 292:297 the glycosylated signature domain 265:297 the glycosylated signature domain of human THBS2 265:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 6 theme signature 282:290 arg1 THBS2 308:312 human THBS2 302:312 human THBS2 302:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 7 theme glycosylated 269:280 arg1 domain 292:297 the glycosylated signature domain 265:297 the glycosylated signature domain of human THBS2 265:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 7 theme glycosylated 269:280 arg1 THBS2 308:312 human THBS2 302:312 human THBS2 302:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 4 8 theme ions 628:631 arg1 network 601:607 a network 599:607 a network of 30 bound Ca(2+) ions and 18 disulfide bonds 599:654 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 16186819 4 9 theme disulfide 640:648 arg1 bonds 650:654 18 disulfide bonds 637:654 18 disulfide bonds 637:654 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 16186819 2 10 theme aspartate-rich 377:390 arg1 modules 365:371 three epidermal growth factor-like modules 330:371 three epidermal growth factor-like modules 330:371 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 10 theme aspartate-rich 377:390 arg1 repeats 392:398 13 aspartate-rich repeats 374:398 13 aspartate-rich repeats 374:398 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 0 11 theme calcium-rich 17:28 arg1 domain 40:45 the calcium-rich signature domain 13:45 the calcium-rich signature domain of human thrombospondin-2 13:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 0 11 theme calcium-rich 17:28 arg1 thrombospondin-2 56:71 human thrombospondin-2 50:71 human thrombospondin-2 50:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 2 12 theme THBS2 308:312 arg1 domain 292:297 the glycosylated signature domain 265:297 the glycosylated signature domain of human THBS2 265:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 12 theme THBS2 308:312 arg1 THBS2 308:312 human THBS2 302:312 human THBS2 302:312 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 3 13 theme structural 476:485 arg1 regions 487:493 three structural regions 470:493 three structural regions termed the stalk, wire and globe 470:526 These elements interact extensively to form three structural regions termed the stalk, wire and globe. 16186819 1 14 gly glycoproteins 111:123 arg1 Thrombospondins 74:88 Thrombospondins 74:88 Thrombospondins (THBSs) 74:96 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 1 14 gly glycoproteins 111:123 arg1 glycoproteins 111:123 secreted glycoproteins 102:123 secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix 102:202 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 1 15 theme secreted 102:109 arg1 Thrombospondins 74:88 Thrombospondins 74:88 Thrombospondins (THBSs) 74:96 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 1 15 theme secreted 102:109 arg1 glycoproteins 111:123 secreted glycoproteins 102:123 secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix 102:202 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 4 16 theme Ca 621:622 arg1 ions 628:631 30 bound Ca(2+) ions 612:631 30 bound Ca(2+) ions 612:631 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 16186819 0 17 theme domain 40:45 arg1 Structure 0:8 Structure 0:8 Structure of the calcium-rich signature domain of human thrombospondin-2 0:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 5 18 theme genetic 684:690 arg1 alterations 692:702 genetic alterations 684:702 genetic alterations of THBSs 684:711 The structure suggests how genetic alterations of THBSs result in disease. 16186819 2 19 theme 2.6-A-resolution 227:242 arg1 structure 252:260 the 2.6-A-resolution crystal structure 223:260 the 2.6-A-resolution crystal structure 223:260 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 1 20 theme extracellular 183:195 arg1 matrix 197:202 the extracellular matrix 179:202 the extracellular matrix 179:202 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 0 21 theme signature 30:38 arg1 domain 40:45 the calcium-rich signature domain 13:45 the calcium-rich signature domain of human thrombospondin-2 13:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 0 21 theme signature 30:38 arg1 thrombospondin-2 56:71 human thrombospondin-2 50:71 human thrombospondin-2 50:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 4 22 theme THBS2 533:537 arg1 domain 549:554 The THBS2 signature domain 529:554 The THBS2 signature domain 529:554 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 16186819 2 23 theme lectin-like 406:416 arg1 module 418:423 a lectin-like module 404:423 a lectin-like module 404:423 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 23 theme lectin-like 406:416 arg1 modules 365:371 three epidermal growth factor-like modules 330:371 three epidermal growth factor-like modules 330:371 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 0 24 theme human 50:54 arg1 thrombospondin-2 56:71 human thrombospondin-2 50:71 human thrombospondin-2 50:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 2 25 theme growth 346:351 arg1 modules 365:371 three epidermal growth factor-like modules 330:371 three epidermal growth factor-like modules 330:371 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 25 theme growth 346:351 arg1 repeats 392:398 13 aspartate-rich repeats 374:398 13 aspartate-rich repeats 374:398 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 25 theme growth 346:351 arg1 module 418:423 a lectin-like module 404:423 a lectin-like module 404:423 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 26 theme epidermal 336:344 arg1 modules 365:371 three epidermal growth factor-like modules 330:371 three epidermal growth factor-like modules 330:371 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 26 theme epidermal 336:344 arg1 repeats 392:398 13 aspartate-rich repeats 374:398 13 aspartate-rich repeats 374:398 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 2 26 theme epidermal 336:344 arg1 module 418:423 a lectin-like module 404:423 a lectin-like module 404:423 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 4 27 theme signature 539:547 arg1 domain 549:554 The THBS2 signature domain 529:554 The THBS2 signature domain 529:554 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 16186819 1 28 theme key 135:137 arg1 roles 139:143 key roles 135:143 key roles 135:143 Thrombospondins (THBSs) are secreted glycoproteins that have key roles in interactions between cells and the extracellular matrix. 16186819 4 29 theme bound 615:619 arg1 ions 628:631 30 bound Ca(2+) ions 612:631 30 bound Ca(2+) ions 612:631 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 16186819 0 30 theme thrombospondin-2 56:71 arg1 domain 40:45 the calcium-rich signature domain 13:45 the calcium-rich signature domain of human thrombospondin-2 13:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 0 30 theme thrombospondin-2 56:71 arg1 thrombospondin-2 56:71 human thrombospondin-2 50:71 human thrombospondin-2 50:71 Structure of the calcium-rich signature domain of human thrombospondin-2. 16186819 2 31 theme crystal 244:250 arg1 structure 252:260 the 2.6-A-resolution crystal structure 223:260 the 2.6-A-resolution crystal structure 223:260 Here, we describe the 2.6-A-resolution crystal structure of the glycosylated signature domain of human THBS2, which includes three epidermal growth factor-like modules, 13 aspartate-rich repeats and a lectin-like module. 16186819 4 32 theme bonds 650:654 arg1 network 601:607 a network 599:607 a network of 30 bound Ca(2+) ions and 18 disulfide bonds 599:654 The THBS2 signature domain is stabilized by these interactions and by a network of 30 bound Ca(2+) ions and 18 disulfide bonds. 12667445 6 0 theme activin 916:922 arg1 receptors 924:932 BMP and activin receptors 908:932 receptors 924:932 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 1 1 theme type 256:259 arg1 receptors 275:283 structurally related type I and type II receptors 235:283 structurally related type I and type II receptors 235:283 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 0 2 theme cooperative 76:86 arg1 nature 88:93 the cooperative nature 72:93 the cooperative nature of receptor assembly 72:114 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 0 3 theme receptor 98:105 arg1 assembly 107:114 receptor assembly 98:114 receptor assembly 98:114 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 2 4 from structure 313:321 arg1 complex 334:340 complex 334:340 complex with the extracellular domain (ECD) of the activin type II receptor 334:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 3 5 theme receptor 502:509 arg1 ECDs 511:514 receptor ECDs 502:514 receptor ECDs 502:514 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 6 6 theme receptors 924:932 arg1 assembly 896:903 the ligand-mediated cooperative assembly 864:903 the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts 864:981 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 3 7 theme direct 524:529 arg1 contacts 531:538 no direct contacts 521:538 no direct contacts 521:538 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 1 8 theme type 267:270 arg1 receptors 275:283 structurally related type I and type II receptors 235:283 structurally related type I and type II receptors 235:283 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 3 9 theme compelling 436:445 arg1 model 461:465 a compelling four-receptor model 434:465 a compelling four-receptor model 434:465 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 5 10 theme ECD 813:815 arg1 presence 780:787 the presence 776:787 the presence of its type II receptor ECD 776:815 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 0 11 theme assembly 107:114 arg1 nature 88:93 the cooperative nature 72:93 the cooperative nature of receptor assembly 72:114 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 1 12 theme diverse 172:178 arg1 responses 191:199 diverse biological responses 172:199 diverse biological responses 172:199 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 0 13 theme BMP7/ActRII 4:14 arg1 complex 37:43 The BMP7/ActRII extracellular domain complex 0:43 The BMP7/ActRII extracellular domain complex 0:43 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 5 14 theme low-affinity 723:734 arg1 receptor 743:750 its low-affinity type I receptor 719:750 its low-affinity type I receptor ECD 719:754 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 2 15 theme crystal 305:311 arg1 structure 313:321 the crystal structure 301:321 the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor 301:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 1 16 theme biological 180:189 arg1 responses 191:199 diverse biological responses 172:199 diverse biological responses 172:199 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 2 17 theme receptor 401:408 arg1 ECD 373:375 ECD 373:375 ECD 373:375 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 17 theme receptor 401:408 arg1 domain 365:370 the extracellular domain 347:370 the extracellular domain (ECD) of the activin type II receptor 347:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 17 theme receptor 401:408 arg1 receptor 401:408 the activin type II receptor 381:408 the activin type II receptor 381:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 1 18 theme receptors 275:283 arg1 pairs 226:230 two pairs 222:230 two pairs of structurally related type I and type II receptors 222:283 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 3 19 dep types 487:491 arg1 types 487:491 the types I and II receptor ECDs 483:514 the types I and II receptor ECDs 483:514 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 3 19 dep types 487:491 arg1 II 499:500 II 499:500 II 499:500 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 3 19 dep types 487:491 arg1 I 493:493 I 493:493 I 493:493 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 2 20 theme type 393:396 arg1 receptor 401:408 the activin type II receptor 381:408 the activin type II receptor 381:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 0 21 theme extracellular 16:28 arg1 complex 37:43 The BMP7/ActRII extracellular domain complex 0:43 The BMP7/ActRII extracellular domain complex 0:43 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 3 22 dep I 493:493 arg1 ECDs 511:514 receptor ECDs 502:514 receptor ECDs 502:514 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 2 23 theme activin 385:391 arg1 receptor 401:408 the activin type II receptor 381:408 the activin type II receptor 381:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 5 24 theme type 736:739 arg1 receptor 743:750 its low-affinity type I receptor 719:750 its low-affinity type I receptor ECD 719:754 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 4 25 theme cell 673:676 arg1 membrane 678:685 the cell membrane 669:685 the cell membrane 669:685 Nevertheless, we find that truncated receptors lacking their cytoplasmic domain retain the ability to cooperatively assemble in the cell membrane. 12667445 6 26 theme assembly 896:903 arg1 view 856:859 a view 854:859 a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts 854:981 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 6 27 theme cooperative 884:894 arg1 assembly 896:903 the ligand-mediated cooperative assembly 864:903 the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts 864:981 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 0 28 theme domain 30:35 arg1 complex 37:43 The BMP7/ActRII extracellular domain complex 0:43 The BMP7/ActRII extracellular domain complex 0:43 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 4 29 theme cytoplasmic 602:612 arg1 domain 614:619 their cytoplasmic domain 596:619 their cytoplasmic domain 596:619 Nevertheless, we find that truncated receptors lacking their cytoplasmic domain retain the ability to cooperatively assemble in the cell membrane. 12667445 5 30 theme receptor 743:750 arg1 ECD 752:754 its low-affinity type I receptor ECD 719:754 its low-affinity type I receptor ECD 719:754 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 6 31 theme ligand-mediated 868:882 arg1 assembly 896:903 the ligand-mediated cooperative assembly 864:903 the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts 864:981 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 6 32 theme BMP 908:910 arg1 receptors 924:932 BMP and activin receptors 908:932 receptors 924:932 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 0 33 theme new 54:56 arg1 insights 58:65 new insights 54:65 new insights into the cooperative nature of receptor assembly 54:114 The BMP7/ActRII extracellular domain complex provides new insights into the cooperative nature of receptor assembly. 12667445 2 34 theme BMP7 326:329 arg1 structure 313:321 the crystal structure 301:321 the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor 301:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 35 with complex 334:340 arg1 ECD 373:375 ECD 373:375 ECD 373:375 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 35 with complex 334:340 arg1 domain 365:370 the extracellular domain 347:370 the extracellular domain (ECD) of the activin type II receptor 347:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 35 with complex 334:340 arg1 receptor 401:408 the activin type II receptor 381:408 the activin type II receptor 381:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 1 36 theme bone 130:133 arg1 BMPs 159:162 BMPs 159:162 BMPs 159:162 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 1 36 theme bone 130:133 arg1 proteins 149:156 bone morphogenetic proteins 130:156 bone morphogenetic proteins (BMPs) 130:163 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 6 37 theme receptor-receptor 956:972 arg1 contacts 974:981 receptor-receptor contacts 956:981 receptor-receptor contacts 956:981 Taken together, our results provide a view of the ligand-mediated cooperative assembly of BMP and activin receptors that does not rely on receptor-receptor contacts. 12667445 1 38 theme morphogenetic 135:147 arg1 BMPs 159:162 BMPs 159:162 BMPs 159:162 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 1 38 theme morphogenetic 135:147 arg1 proteins 149:156 bone morphogenetic proteins 130:156 bone morphogenetic proteins (BMPs) 130:163 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12667445 3 39 theme four-receptor 447:459 arg1 model 461:465 a compelling four-receptor model 434:465 a compelling four-receptor model 434:465 Our structure produces a compelling four-receptor model, revealing that the types I and II receptor ECDs make no direct contacts. 12667445 5 40 theme BMP7 710:713 arg1 affinity 698:705 the affinity 694:705 the affinity of BMP7 for its low-affinity type I receptor ECD 694:754 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 5 41 theme type 796:799 arg1 receptor 804:811 type II receptor 796:811 its type II receptor ECD 792:815 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 2 42 theme extracellular 351:363 arg1 ECD 373:375 ECD 373:375 ECD 373:375 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 42 theme extracellular 351:363 arg1 domain 365:370 the extracellular domain 347:370 the extracellular domain (ECD) of the activin type II receptor 347:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 2 42 theme extracellular 351:363 arg1 receptor 401:408 the activin type II receptor 381:408 the activin type II receptor 381:408 Here we report the crystal structure of BMP7 in complex with the extracellular domain (ECD) of the activin type II receptor. 12667445 5 43 theme receptor 804:811 arg1 ECD 813:815 its type II receptor ECD 792:815 its type II receptor ECD 792:815 Also, the affinity of BMP7 for its low-affinity type I receptor ECD increases 5-fold in the presence of its type II receptor ECD. 12667445 4 44 theme truncated 568:576 arg1 receptors 578:586 truncated receptors 568:586 truncated receptors lacking their cytoplasmic domain 568:619 Nevertheless, we find that truncated receptors lacking their cytoplasmic domain retain the ability to cooperatively assemble in the cell membrane. 12667445 1 45 theme related 248:254 arg1 receptors 275:283 structurally related type I and type II receptors 235:283 structurally related type I and type II receptors 235:283 Activins and bone morphogenetic proteins (BMPs) elicit diverse biological responses by signaling through two pairs of structurally related type I and type II receptors. 12478285 3 0 theme epitopes 683:690 arg1 interfaces 657:666 the molecular interfaces 643:666 the molecular interfaces of the binding epitopes for both type I and type II receptors 643:728 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 4 1 theme BMP-7-binding 735:747 arg1 affinity 749:756 The BMP-7-binding affinity 731:756 The BMP-7-binding affinity of site-specific variants of Noggin 731:792 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 1 2 theme embryonic 241:249 arg1 axis 266:269 the embryonic dorsal-ventral axis 237:269 the embryonic dorsal-ventral axis 237:269 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 0 3 theme protein 66:72 arg1 Noggin 74:79 the cystine knot protein Noggin 49:79 the cystine knot protein Noggin 49:79 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 1 4 theme skeletal 327:334 arg1 system 336:341 the skeletal system 323:341 the skeletal system 323:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 5 from joints 313:318 arg1 system 336:341 the skeletal system 323:341 the skeletal system 323:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 6 theme dorsal-ventral 251:264 arg1 axis 266:269 the embryonic dorsal-ventral axis 237:269 the embryonic dorsal-ventral axis 237:269 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 5 7 theme similar 1053:1059 arg1 topology 1044:1051 a cystine (the oxidized form of cysteine) knot topology 997:1051 a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs 997:1075 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 5 8 theme ancestral 1141:1149 arg1 gene 1151:1154 a common ancestral gene 1132:1154 a common ancestral gene 1132:1154 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 4 9 theme chick 860:864 arg1 development 871:881 chick limb development 860:881 chick limb development 860:881 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 3 10 theme crystal 524:530 arg1 structure 532:540 the crystal structure 520:540 the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors 520:728 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 1 11 theme axis 266:269 arg1 neurogenesis 347:358 neurogenesis 347:358 neurogenesis in the adult brain 347:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 11 theme axis 266:269 arg1 induction 272:280 induction 272:280 induction of neural tissue 272:297 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 11 theme axis 266:269 arg1 establishment 220:232 establishment 220:232 establishment of the embryonic dorsal-ventral axis 220:269 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 11 theme axis 266:269 arg1 formation 300:308 formation 300:308 formation of joints in the skeletal system 300:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 4 12 from formation 833:841 arg1 development 871:881 chick limb development 860:881 chick limb development 860:881 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 1 13 from formation 300:308 arg1 system 336:341 the skeletal system 323:341 the skeletal system 323:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 13 from formation 300:308 arg1 brain 373:377 the adult brain 363:377 the adult brain 363:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 5 14 dep cystine 999:1005 arg1 form 1021:1024 the oxidized form 1008:1024 the oxidized form of cysteine 1008:1036 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 2 15 theme BMP 424:426 arg1 antagonists 428:438 BMP antagonists 424:438 BMP antagonists 424:438 So far, the three-dimensional structures of BMP antagonists and the structural basis for inactivation have remained unknown. 12478285 2 16 theme antagonists 428:438 arg1 structures 410:419 the three-dimensional structures 388:419 the three-dimensional structures of BMP antagonists 388:438 So far, the three-dimensional structures of BMP antagonists and the structural basis for inactivation have remained unknown. 12478285 2 16 theme antagonists 428:438 arg1 basis 459:463 the structural basis 444:463 the structural basis for inactivation 444:480 So far, the three-dimensional structures of BMP antagonists and the structural basis for inactivation have remained unknown. 12478285 4 17 from alterations 813:823 arg1 apoptosis 847:855 apoptosis 847:855 apoptosis 847:855 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 4 17 from alterations 813:823 arg1 formation 833:841 bone formation 828:841 bone formation 828:841 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 1 18 from induction 272:280 arg1 system 336:341 the skeletal system 323:341 the skeletal system 323:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 18 from induction 272:280 arg1 brain 373:377 the adult brain 363:377 the adult brain 363:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 4 19 theme inactive 947:954 arg1 complex 956:962 an inactive complex 944:962 an inactive complex 944:962 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 3 20 theme antagonist 549:558 arg1 Noggin 560:565 the antagonist Noggin 545:565 the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors 545:728 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 1 21 theme developmental 169:181 arg1 processes 196:204 developmental and cellular processes 169:204 developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain 169:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 0 22 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 0:80 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 4 23 theme bone 828:831 arg1 formation 833:841 bone formation 828:841 bone formation 828:841 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 3 24 theme Noggin 560:565 arg1 structure 532:540 the crystal structure 520:540 the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors 520:728 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 0 25 theme BMP 20:22 arg1 inhibition 35:44 BMP signalling inhibition 20:44 BMP signalling inhibition 20:44 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 1 26 theme cellular 187:194 arg1 processes 196:204 developmental and cellular processes 169:204 developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain 169:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 27 from neurogenesis 347:358 arg1 system 336:341 the skeletal system 323:341 the skeletal system 323:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 27 from neurogenesis 347:358 arg1 brain 373:377 the adult brain 363:377 the adult brain 363:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 28 theme neural 285:290 arg1 tissue 292:297 neural tissue 285:297 neural tissue 285:297 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 29 theme adult 367:371 arg1 brain 373:377 the adult brain 363:377 the adult brain 363:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 2 30 theme structural 448:457 arg1 basis 459:463 the structural basis 444:463 the structural basis for inactivation 444:480 So far, the three-dimensional structures of BMP antagonists and the structural basis for inactivation have remained unknown. 12478285 0 31 theme inhibition 35:44 arg1 basis 11:15 Structural basis 0:15 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 0:80 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 1 32 theme tissue 292:297 arg1 neurogenesis 347:358 neurogenesis 347:358 neurogenesis in the adult brain 347:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 32 theme tissue 292:297 arg1 induction 272:280 induction 272:280 induction of neural tissue 272:297 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 32 theme tissue 292:297 arg1 establishment 220:232 establishment 220:232 establishment of the embryonic dorsal-ventral axis 220:269 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 32 theme tissue 292:297 arg1 formation 300:308 formation 300:308 formation of joints in the skeletal system 300:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 5 33 theme Noggin 981:986 arg1 scaffold 969:976 The scaffold 965:976 The scaffold of Noggin 965:986 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 4 34 from ligand 934:939 arg1 complex 956:962 an inactive complex 944:962 an inactive complex 944:962 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 1 35 from system 336:341 arg1 neurogenesis 347:358 neurogenesis 347:358 neurogenesis in the adult brain 347:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 35 from system 336:341 arg1 induction 272:280 induction 272:280 induction of neural tissue 272:297 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 35 from system 336:341 arg1 establishment 220:232 establishment 220:232 establishment of the embryonic dorsal-ventral axis 220:269 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 35 from system 336:341 arg1 formation 300:308 formation 300:308 formation of joints in the skeletal system 300:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 0 36 theme signalling 24:33 arg1 inhibition 35:44 BMP signalling inhibition 20:44 BMP signalling inhibition 20:44 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 5 37 theme knot 1039:1042 arg1 topology 1044:1051 a cystine (the oxidized form of cysteine) knot topology 997:1051 a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs 997:1075 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 4 38 theme Noggin 787:792 arg1 variants 775:782 site-specific variants 761:782 site-specific variants of Noggin 761:792 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 1 39 theme diverse 209:215 arg1 processes 196:204 developmental and cellular processes 169:204 developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain 169:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 5 40 theme cysteine 1029:1036 arg1 form 1021:1024 the oxidized form 1008:1024 the oxidized form of cysteine 1008:1036 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 3 41 theme BMP 616:618 arg1 signalling 620:629 BMP signalling 616:629 BMP signalling 616:629 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 5 42 theme cystine 999:1005 arg1 topology 1044:1051 a cystine (the oxidized form of cysteine) knot topology 997:1051 a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs 997:1075 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 1 43 theme bone 104:107 arg1 proteins 123:130 bone morphogenetic proteins 104:130 bone morphogenetic proteins (BMPs) 104:137 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 43 theme bone 104:107 arg1 BMPs 133:136 BMPs 133:136 BMPs 133:136 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 3 44 dep type 701:704 arg1 receptors 720:728 receptors 720:728 receptors 720:728 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 4 45 theme limb 866:869 arg1 development 871:881 chick limb development 860:881 chick limb development 860:881 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 3 46 theme molecular 647:655 arg1 interfaces 657:666 the molecular interfaces 643:666 the molecular interfaces of the binding epitopes for both type I and type II receptors 643:728 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 1 47 theme morphogenetic 109:121 arg1 proteins 123:130 bone morphogenetic proteins 104:130 bone morphogenetic proteins (BMPs) 104:137 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 47 theme morphogenetic 109:121 arg1 BMPs 133:136 BMPs 133:136 BMPs 133:136 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 48 theme joints 313:318 arg1 neurogenesis 347:358 neurogenesis 347:358 neurogenesis in the adult brain 347:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 48 theme joints 313:318 arg1 induction 272:280 induction 272:280 induction of neural tissue 272:297 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 48 theme joints 313:318 arg1 establishment 220:232 establishment 220:232 establishment of the embryonic dorsal-ventral axis 220:269 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 48 theme joints 313:318 arg1 formation 300:308 formation 300:308 formation of joints in the skeletal system 300:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 5 49 theme oxidized 1012:1019 arg1 form 1021:1024 the oxidized form 1008:1024 the oxidized form of cysteine 1008:1036 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 4 50 theme site-specific 761:773 arg1 variants 775:782 site-specific variants 761:782 site-specific variants of Noggin 761:792 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 1 51 from establishment 220:232 arg1 system 336:341 the skeletal system 323:341 the skeletal system 323:341 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 1 51 from establishment 220:232 arg1 brain 373:377 the adult brain 363:377 the adult brain 363:377 The interplay between bone morphogenetic proteins (BMPs) and their antagonists governs developmental and cellular processes as diverse as establishment of the embryonic dorsal-ventral axis, induction of neural tissue, formation of joints in the skeletal system and neurogenesis in the adult brain. 12478285 4 52 from apoptosis 847:855 arg1 development 871:881 chick limb development 860:881 chick limb development 860:881 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 0 53 theme knot 61:64 arg1 Noggin 74:79 the cystine knot protein Noggin 49:79 the cystine knot protein Noggin 49:79 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 2 54 theme three-dimensional 392:408 arg1 structures 410:419 the three-dimensional structures 388:419 the three-dimensional structures of BMP antagonists 388:438 So far, the three-dimensional structures of BMP antagonists and the structural basis for inactivation have remained unknown. 12478285 5 55 contain contains 988:995 arg2 topology 1044:1051 a cystine (the oxidized form of cysteine) knot topology 997:1051 a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs 997:1075 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 5 55 contain contains 988:995 arg1 scaffold 969:976 The scaffold 965:976 The scaffold of Noggin 965:986 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 12478285 0 56 theme cystine 53:59 arg1 Noggin 74:79 the cystine knot protein Noggin 49:79 the cystine knot protein Noggin 49:79 Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. 12478285 3 57 theme binding 675:681 arg1 epitopes 683:690 the binding epitopes 671:690 the binding epitopes 671:690 Here we report the crystal structure of the antagonist Noggin bound to BMP-7, which shows that Noggin inhibits BMP signalling by blocking the molecular interfaces of the binding epitopes for both type I and type II receptors. 12478285 4 58 theme variants 775:782 arg1 affinity 749:756 The BMP-7-binding affinity 731:756 The BMP-7-binding affinity of site-specific variants of Noggin 731:792 The BMP-7-binding affinity of site-specific variants of Noggin is correlated with alterations in bone formation and apoptosis in chick limb development, showing that Noggin functions by sequestering its ligand in an inactive complex. 12478285 5 59 theme common 1134:1139 arg1 gene 1151:1154 a common ancestral gene 1132:1154 a common ancestral gene 1132:1154 The scaffold of Noggin contains a cystine (the oxidized form of cysteine) knot topology similar to that of BMPs; thus, ligand and antagonist seem to have evolved from a common ancestral gene. 16870210 2 0 from specificity 486:496 arg1 restricted 466:475 restricted 466:475 restricted 466:475 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 7 1 theme hormone 1484:1490 arg1 side-chains 1492:1502 hormone side-chains 1484:1502 hormone side-chains 1484:1502 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 6 2 theme CNP 1183:1185 arg1 structure 1187:1195 the CNP structure 1179:1195 the CNP structure 1179:1195 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 3 3 theme brain 741:745 arg1 BNP 768:770 BNP 768:770 BNP 768:770 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 3 theme brain 741:745 arg1 peptide 759:765 brain natriuretic peptide 741:765 brain natriuretic peptide (BNP) 741:771 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 6 4 theme other 1260:1264 arg1 ligands 1266:1272 the other ligands 1256:1272 the other ligands 1256:1272 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 4 5 theme structural 776:785 arg1 comparison 787:796 A structural comparison 774:796 A structural comparison 774:796 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 4 6 dep different 948:956 arg1 NP 976:977 NP 976:977 NP 976:977 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 4 6 dep different 948:956 arg1 flexible 966:973 flexible 966:973 flexible 966:973 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 7 7 theme different 1341:1349 arg1 NPRs 1351:1354 different NPRs 1341:1354 different NPRs 1341:1354 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 8 theme selective 1297:1305 arg1 properties 1327:1336 The degenerate versus selective hormone recognition properties 1275:1336 The degenerate versus selective hormone recognition properties of different NPRs 1275:1354 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 3 9 theme crystal 648:654 arg1 structures 656:665 the crystal structures 644:665 the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP) 644:771 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 4 10 theme inflexible 915:924 arg1 surface 926:932 a conformationally inflexible surface 896:932 a conformationally inflexible surface 896:932 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 1 11 from part 157:160 arg1 reliant 145:151 reliant 145:151 reliant 145:151 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 7 12 theme NPRs 1351:1354 arg1 properties 1327:1336 The degenerate versus selective hormone recognition properties 1275:1336 The degenerate versus selective hormone recognition properties of different NPRs 1275:1354 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 4 13 theme NPR-C/CNP 853:861 arg1 complex 863:869 the NPR-C/CNP complex 849:869 the NPR-C/CNP complex 849:869 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 1 14 theme Cardiovascular 84:97 arg1 homeostasis 99:109 Cardiovascular homeostasis 84:109 Cardiovascular homeostasis 84:109 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 2 15 theme other 439:443 arg1 NPRs 445:448 other NPRs 439:448 other NPRs 439:448 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 5 16 theme complex 992:998 arg1 structures 1000:1009 The complex structures 988:1009 The complex structures 988:1009 The complex structures support a mechanism of rigid promiscuity rather than conformational plasticity by the receptor. 16870210 6 17 theme receptor-bound 1149:1162 arg1 conformations 1164:1176 similar receptor-bound conformations 1141:1176 similar receptor-bound conformations 1141:1176 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 2 18 theme C-type 266:271 arg1 NPR-C 278:282 NPR-C 278:282 NPR-C 278:282 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 18 theme C-type 266:271 arg1 responsible 288:298 responsible 288:298 responsible 288:298 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 18 theme C-type 266:271 arg1 NPR 273:275 The C-type NPR 262:275 The C-type NPR (NPR-C) 262:283 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 1 19 theme natriuretic 187:197 arg1 peptide 199:205 natriuretic peptide 187:205 natriuretic peptide (NP) hormones 187:219 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 1 19 theme natriuretic 187:197 arg1 NP 208:209 NP 208:209 NP 208:209 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 6 20 theme similar 1141:1147 arg1 conformations 1164:1176 similar receptor-bound conformations 1141:1176 similar receptor-bound conformations 1141:1176 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 5 21 theme conformational 1064:1077 arg1 plasticity 1079:1088 conformational plasticity 1064:1088 a mechanism of rigid promiscuity rather than conformational plasticity by the receptor 1019:1104 The complex structures support a mechanism of rigid promiscuity rather than conformational plasticity by the receptor. 16870210 7 22 theme hormone 1307:1313 arg1 properties 1327:1336 The degenerate versus selective hormone recognition properties 1275:1336 The degenerate versus selective hormone recognition properties of different NPRs 1275:1354 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 1 23 theme peptide 199:205 arg1 hormones 212:219 natriuretic peptide (NP) hormones 187:219 natriuretic peptide (NP) hormones 187:219 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 0 24 theme Structural 0:9 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of natriuretic peptide 0:45 Structural determinants of natriuretic peptide receptor specificity and degeneracy. 16870210 3 25 theme complexes 674:682 arg1 structures 656:665 the crystal structures 644:665 the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP) 644:771 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 7 26 theme receptor 1407:1414 arg1 surfaces 1416:1423 the receptor surfaces 1403:1423 the receptor surfaces 1403:1423 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 1 27 theme blood 115:119 arg1 regulation 130:139 blood pressure regulation 115:139 blood pressure regulation 115:139 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 0 28 theme natriuretic 27:37 arg1 peptide 39:45 natriuretic peptide 27:45 natriuretic peptide 27:45 Structural determinants of natriuretic peptide receptor specificity and degeneracy. 16870210 6 29 theme receptor 1233:1240 arg1 contacts 1242:1249 common receptor contacts 1226:1249 common receptor contacts 1226:1249 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 2 30 from circulation 338:348 arg1 clearance 304:312 clearance 304:312 clearance of NP hormones from the circulation 304:348 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 1 31 theme pressure 121:128 arg1 regulation 130:139 blood pressure regulation 115:139 blood pressure regulation 115:139 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 3 32 with specificity 565:575 arg1 hormones 615:622 NP hormones 612:622 NP hormones 612:622 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 7 33 theme recognition 1315:1325 arg1 properties 1327:1336 The degenerate versus selective hormone recognition properties 1275:1336 The degenerate versus selective hormone recognition properties of different NPRs 1275:1354 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 34 theme receptor 1517:1524 arg1 selectivity 1526:1536 receptor selectivity 1517:1536 receptor selectivity 1517:1536 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 34 theme receptor 1517:1524 arg1 sites 1474:1478 anchoring sites 1464:1478 anchoring sites for hormone side-chains 1464:1502 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 34 theme receptor 1517:1524 arg1 cavities 1391:1398 two cavities 1387:1398 two cavities on the receptor surfaces 1387:1423 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 6 35 theme common 1226:1231 arg1 contacts 1242:1249 common receptor contacts 1226:1249 common receptor contacts 1226:1249 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 4 36 with comparison 787:796 arg1 structure 836:844 the previous structure 823:844 the previous structure of the NPR-C/CNP complex 823:869 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 2 37 theme NP 391:392 arg1 BNP 409:411 BNP 409:411 BNP 409:411 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 37 theme NP 391:392 arg1 CNP 418:420 CNP 418:420 CNP 418:420 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 37 theme NP 391:392 arg1 ANP 404:406 ANP 404:406 ANP 404:406 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 37 theme NP 391:392 arg1 hormones 394:401 all NP hormones 387:401 all NP hormones (ANP, BNP, and CNP) 387:421 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 4 38 theme previous 827:834 arg1 structure 836:844 the previous structure 823:844 the previous structure of the NPR-C/CNP complex 823:869 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 7 39 theme pocket 1439:1444 arg1 II 1446:1447 pocket II 1439:1447 pocket II 1439:1447 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 39 theme pocket 1439:1444 arg1 cavities 1391:1398 two cavities 1387:1398 two cavities on the receptor surfaces 1387:1423 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 3 40 with cross-reactivity 581:596 arg1 hormones 615:622 NP hormones 612:622 NP hormones 612:622 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 0 41 theme peptide 39:45 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of natriuretic peptide 0:45 Structural determinants of natriuretic peptide receptor specificity and degeneracy. 16870210 3 42 theme atrial 698:703 arg1 ANP 726:728 ANP 726:728 ANP 726:728 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 42 theme atrial 698:703 arg1 peptide 717:723 atrial natriuretic peptide 698:723 atrial natriuretic peptide (ANP) 698:729 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 43 theme binding 557:563 arg1 specificity 565:575 the binding specificity 553:575 the binding specificity 553:575 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 7 44 theme degenerate 1279:1288 arg1 properties 1327:1336 The degenerate versus selective hormone recognition properties 1275:1336 The degenerate versus selective hormone recognition properties of different NPRs 1275:1354 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 6 45 theme contacts 1242:1249 arg1 contacts 1242:1249 common receptor contacts 1226:1249 common receptor contacts 1226:1249 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 6 45 theme contacts 1242:1249 arg1 sets 1218:1221 sets 1218:1221 sets of common receptor contacts 1218:1249 While ANP and BNP appear to adopt similar receptor-bound conformations, the CNP structure diverges, yet shares sets of common receptor contacts with the other ligands. 16870210 7 46 from cavities 1391:1398 arg1 surfaces 1416:1423 the receptor surfaces 1403:1423 the receptor surfaces 1403:1423 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 3 47 theme natriuretic 705:715 arg1 ANP 726:728 ANP 726:728 ANP 726:728 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 47 theme natriuretic 705:715 arg1 peptide 717:723 atrial natriuretic peptide 698:723 atrial natriuretic peptide (ANP) 698:729 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 2 48 dep hormones 394:401 arg1 BNP 409:411 BNP 409:411 BNP 409:411 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 48 dep hormones 394:401 arg1 CNP 418:420 CNP 418:420 CNP 418:420 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 48 dep hormones 394:401 arg1 ANP 404:406 ANP 404:406 ANP 404:406 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 48 dep hormones 394:401 arg1 hormones 394:401 all NP hormones 387:401 all NP hormones (ANP, BNP, and CNP) 387:421 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 2 49 theme hormones 320:327 arg1 clearance 304:312 clearance 304:312 clearance of NP hormones from the circulation 304:348 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 3 50 theme NPR-C 687:691 arg1 complexes 674:682 the complexes 670:682 the complexes of NPR-C 670:691 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 5 51 theme rigid 1034:1038 arg1 promiscuity 1040:1050 rigid promiscuity 1034:1050 rigid promiscuity 1034:1050 The complex structures support a mechanism of rigid promiscuity rather than conformational plasticity by the receptor. 16870210 2 52 theme NP 317:318 arg1 hormones 320:327 NP hormones 317:327 NP hormones 317:327 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 1 53 from reliant 145:151 arg1 part 157:160 part 157:160 part 157:160 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 4 54 theme complexes 807:815 arg1 comparison 787:796 A structural comparison 774:796 A structural comparison 774:796 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 3 55 theme NP 612:613 arg1 hormones 615:622 NP hormones 612:622 NP hormones 612:622 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 1 56 theme natriuretic 225:235 arg1 receptors 245:253 natriuretic peptide receptors 225:253 natriuretic peptide receptors (NPR) 225:259 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 1 56 theme natriuretic 225:235 arg1 NPR 256:258 NPR 256:258 NPR 256:258 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 7 57 theme pocket 1426:1431 arg1 I 1433:1433 pocket I 1426:1433 pocket I 1426:1433 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 57 theme pocket 1426:1431 arg1 cavities 1391:1398 two cavities 1387:1398 two cavities on the receptor surfaces 1387:1423 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 3 58 theme natriuretic 747:757 arg1 BNP 768:770 BNP 768:770 BNP 768:770 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 58 theme natriuretic 747:757 arg1 peptide 759:765 brain natriuretic peptide 741:765 brain natriuretic peptide (BNP) 741:771 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 1 59 theme peptide 237:243 arg1 receptors 245:253 natriuretic peptide receptors 225:253 natriuretic peptide receptors (NPR) 225:259 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 1 59 theme peptide 237:243 arg1 NPR 256:258 NPR 256:258 NPR 256:258 Cardiovascular homeostasis and blood pressure regulation are reliant, in part, on interactions between natriuretic peptide (NP) hormones and natriuretic peptide receptors (NPR). 16870210 3 60 theme NPR-C 601:605 arg1 cross-reactivity 581:596 cross-reactivity 581:596 cross-reactivity of NPR-C with NP hormones 581:622 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 60 theme NPR-C 601:605 arg1 specificity 565:575 the binding specificity 553:575 the binding specificity 553:575 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 61 theme structural 525:534 arg1 determinants 536:547 the structural determinants 521:547 the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones 521:622 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 62 with structures 656:665 arg1 ANP 726:728 ANP 726:728 ANP 726:728 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 62 with structures 656:665 arg1 peptide 717:723 atrial natriuretic peptide 698:723 atrial natriuretic peptide (ANP) 698:729 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 62 with structures 656:665 arg1 BNP 768:770 BNP 768:770 BNP 768:770 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 3 62 with structures 656:665 arg1 peptide 759:765 brain natriuretic peptide 741:765 brain natriuretic peptide (BNP) 741:771 In order to elucidate the structural determinants for the binding specificity and cross-reactivity of NPR-C with NP hormones, we have determined the crystal structures of the complexes of NPR-C with atrial natriuretic peptide (ANP), and with brain natriuretic peptide (BNP). 16870210 4 63 theme complex 863:869 arg1 structure 836:844 the previous structure 823:844 the previous structure of the NPR-C/CNP complex 823:869 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 5 64 theme promiscuity 1040:1050 arg1 mechanism 1021:1029 a mechanism 1019:1029 a mechanism of rigid promiscuity rather than conformational plasticity by the receptor 1019:1104 The complex structures support a mechanism of rigid promiscuity rather than conformational plasticity by the receptor. 16870210 7 65 theme anchoring 1464:1472 arg1 cavities 1391:1398 two cavities 1387:1398 two cavities on the receptor surfaces 1387:1423 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 65 theme anchoring 1464:1472 arg1 sites 1474:1478 anchoring sites 1464:1478 anchoring sites for hormone side-chains 1464:1502 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 7 65 theme anchoring 1464:1472 arg1 selectivity 1526:1536 receptor selectivity 1517:1536 receptor selectivity 1517:1536 The degenerate versus selective hormone recognition properties of different NPRs appears to derive largely from two cavities on the receptor surfaces, pocket I and pocket II, that serve as anchoring sites for hormone side-chains and modulate receptor selectivity. 16870210 2 66 from hormones 394:401 arg1 contrast 427:434 contrast 427:434 contrast to other NPRs 427:448 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 16870210 4 67 theme different 948:956 arg1 ligands 979:985 three different, highly flexible, NP ligands 942:985 three different, highly flexible, NP ligands 942:985 A structural comparison of these complexes, with the previous structure of the NPR-C/CNP complex, reveals that NPR-C uses a conformationally inflexible surface to bind three different, highly flexible, NP ligands. 16870210 2 68 from restricted 466:475 arg1 specificity 486:496 their specificity 480:496 their specificity 480:496 The C-type NPR (NPR-C) is responsible for clearance of NP hormones from the circulation, and displays a cross-reactivity for all NP hormones (ANP, BNP, and CNP), in contrast to other NPRs, which are more restricted in their specificity. 28377511 6 0 theme CRD 870:872 arg1 arrangement 851:861 a dimeric arrangement 841:861 a dimeric arrangement of the CRD 841:872 Both structures reveal a dimeric arrangement of the CRD. 28377511 10 1 theme CRD 1349:1351 arg1 monomers 1353:1360 two CRD monomers 1345:1360 two CRD monomers 1345:1360 The fatty acid bridges two CRD monomers, implying that Wnt binding mediates FZD receptor dimerization. 28377511 2 2 theme pancreatic 333:342 arg1 tumors 378:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 2 3 from signaling 309:317 arg1 subsets 322:328 subsets 322:328 subsets of pancreatic, ovarian, gastric, and colorectal tumors 322:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 11 4 theme CRD 1487:1489 arg1 complexes 1491:1499 Wnt-FZD CRD complexes 1479:1499 Wnt-FZD CRD complexes 1479:1499 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 3 5 theme groups 540:545 arg1 recognition 494:504 recognition 494:504 recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors 494:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 6 6 theme dimeric 843:849 arg1 arrangement 851:861 a dimeric arrangement 841:861 a dimeric arrangement of the CRD 841:872 Both structures reveal a dimeric arrangement of the CRD. 28377511 3 7 theme fatty 529:533 arg1 groups 540:545 Wnt cis-unsaturated fatty acyl groups 509:545 Wnt cis-unsaturated fatty acyl groups 509:545 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 9 8 theme common 1159:1164 arg1 mechanism 1176:1184 a common molecular mechanism 1157:1184 a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors 1157:1319 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 2 9 theme overactive 294:303 arg1 signaling 309:317 overactive Wnt signaling 294:317 overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors 294:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 1 10 theme bone 185:188 arg1 growth 190:195 bone growth 185:195 bone growth 185:195 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 5 11 theme fatty 806:810 arg1 acid 812:815 C16:1 cis-Δ9 unsaturated fatty acid 781:815 C16:1 cis-Δ9 unsaturated fatty acid 781:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 28377511 7 12 theme U-shaped 957:964 arg1 geometry 966:973 a U-shaped geometry 955:973 a U-shaped geometry that accommodates the fatty acid 955:1006 The lipid-binding groove exhibits flexibility and spans both monomers, adopting a U-shaped geometry that accommodates the fatty acid. 28377511 7 13 theme fatty 997:1001 arg1 acid 1003:1006 the fatty acid 993:1006 the fatty acid 993:1006 The lipid-binding groove exhibits flexibility and spans both monomers, adopting a U-shaped geometry that accommodates the fatty acid. 28377511 9 14 theme fatty 1225:1229 arg1 modification 1273:1284 a necessary posttranslational modification 1243:1284 a necessary posttranslational modification of Wnts 1243:1292 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 14 theme fatty 1225:1229 arg1 group 1236:1240 the cis-unsaturated fatty acyl group 1205:1240 the cis-unsaturated fatty acyl group 1205:1240 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 2 15 theme receptor 239:246 arg1 expression 248:257 high FZD receptor expression 230:257 high FZD receptor expression at the cell surface 230:277 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 2 16 from surface 271:277 arg1 expression 248:257 high FZD receptor expression 230:257 high FZD receptor expression at the cell surface 230:277 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 4 17 theme human 658:662 arg1 CRD 669:671 human FZD7 CRD 658:671 human FZD7 CRD 658:671 Here, we determined a crystal structure of human FZD7 CRD unexpectedly bound to a 24-carbon fatty acid. 28377511 9 18 theme group 1236:1240 arg1 recognition 1190:1200 recognition 1190:1200 recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors 1190:1319 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 3 19 theme receptors 588:596 arg1 receptors 588:596 FZD receptors 584:596 FZD receptors 584:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 19 theme receptors 588:596 arg1 CRD 576:578 CRD 576:578 CRD 576:578 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 19 theme receptors 588:596 arg1 domain 568:573 the cysteine-rich domain 550:573 the cysteine-rich domain (CRD) of FZD receptors 550:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 10 20 theme FZD 1398:1400 arg1 dimerization 1411:1422 FZD receptor dimerization 1398:1422 FZD receptor dimerization 1398:1422 The fatty acid bridges two CRD monomers, implying that Wnt binding mediates FZD receptor dimerization. 28377511 11 21 theme Wnt-FZD 1479:1485 arg1 complexes 1491:1499 Wnt-FZD CRD complexes 1479:1499 Wnt-FZD CRD complexes 1479:1499 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 0 22 theme Wnt 84:86 arg1 binding 95:101 Wnt ligand binding 84:101 Wnt ligand binding 84:101 Unsaturated fatty acyl recognition by Frizzled receptors mediates dimerization upon Wnt ligand binding. 28377511 1 23 theme diverse 154:160 arg1 processes 162:170 diverse processes 154:170 diverse processes ranging from bone growth to stem cell activity 154:217 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 8 24 dep FZD5 1118:1121 arg1 CRDs 1132:1135 CRDs 1132:1135 CRDs 1132:1135 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 9 25 theme posttranslational 1255:1271 arg1 modification 1273:1284 a necessary posttranslational modification 1243:1284 a necessary posttranslational modification of Wnts 1243:1292 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 25 theme posttranslational 1255:1271 arg1 group 1236:1240 the cis-unsaturated fatty acyl group 1205:1240 the cis-unsaturated fatty acyl group 1205:1240 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 2 26 theme cell 266:269 arg1 surface 271:277 the cell surface 262:277 the cell surface 262:277 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 2 27 theme colorectal 367:376 arg1 tumors 378:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 0 28 theme Frizzled 38:45 arg1 receptors 47:55 Frizzled receptors 38:55 Frizzled receptors 38:55 Unsaturated fatty acyl recognition by Frizzled receptors mediates dimerization upon Wnt ligand binding. 28377511 4 29 theme fatty 707:711 arg1 acid 713:716 a 24-carbon fatty acid 695:716 a 24-carbon fatty acid 695:716 Here, we determined a crystal structure of human FZD7 CRD unexpectedly bound to a 24-carbon fatty acid. 28377511 5 30 theme CRD 768:770 arg1 structure 744:752 a crystal structure 734:752 a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid 734:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 28377511 2 31 theme FZD 235:237 arg1 expression 248:257 high FZD receptor expression 230:257 high FZD receptor expression at the cell surface 230:277 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 3 32 theme receptor 447:454 arg1 interactions 456:467 Wnt-FZD receptor interactions 439:467 Wnt-FZD receptor interactions 439:467 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 0 33 theme acyl 18:21 arg1 recognition 23:33 Unsaturated fatty acyl recognition 0:33 Unsaturated fatty acyl recognition by Frizzled receptors 0:55 Unsaturated fatty acyl recognition by Frizzled receptors mediates dimerization upon Wnt ligand binding. 28377511 9 34 theme multiple 1298:1305 arg1 receptors 1311:1319 multiple FZD receptors 1298:1319 multiple FZD receptors 1298:1319 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 8 35 theme mouse 1040:1044 arg1 structure 1055:1063 the published mouse FZD8 CRD structure 1026:1063 the published mouse FZD8 CRD structure 1026:1063 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 8 36 theme CRD 1051:1053 arg1 structure 1055:1063 the published mouse FZD8 CRD structure 1026:1063 the published mouse FZD8 CRD structure 1026:1063 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 3 37 theme molecular 474:482 arg1 basis 484:488 the molecular basis 470:488 the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors 470:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 5 38 theme human 757:761 arg1 CRD 768:770 human FZD5 CRD 757:770 human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid 757:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 28377511 2 39 theme high 230:233 arg1 expression 248:257 high FZD receptor expression 230:257 high FZD receptor expression at the cell surface 230:277 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 2 40 theme tumors 378:383 arg1 subsets 322:328 subsets 322:328 subsets of pancreatic, ovarian, gastric, and colorectal tumors 322:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 11 41 theme pharmacological 1571:1585 arg1 strategies 1587:1596 pharmacological strategies to modulate FZD receptor function 1571:1630 pharmacological strategies to modulate FZD receptor function 1571:1630 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 8 42 theme same 1097:1100 arg1 architecture 1102:1113 the same architecture 1093:1113 the same architecture 1093:1113 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 11 43 theme FZD 1610:1612 arg1 function 1623:1630 FZD receptor function 1610:1630 FZD receptor function 1610:1630 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 3 44 theme acyl 535:538 arg1 groups 540:545 Wnt cis-unsaturated fatty acyl groups 509:545 Wnt cis-unsaturated fatty acyl groups 509:545 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 2 45 theme Wnt 305:307 arg1 signaling 309:317 overactive Wnt signaling 294:317 overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors 294:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 11 46 theme complexes 1491:1499 arg1 arrangement 1464:1474 the arrangement 1460:1474 the arrangement of Wnt-FZD CRD complexes 1460:1499 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 3 47 theme cis-unsaturated 513:527 arg1 groups 540:545 Wnt cis-unsaturated fatty acyl groups 509:545 Wnt cis-unsaturated fatty acyl groups 509:545 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 48 from progress 398:405 arg1 understanding 422:434 biochemical understanding 410:434 biochemical understanding of Wnt-FZD receptor interactions 410:467 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 49 theme cysteine-rich 554:566 arg1 receptors 588:596 FZD receptors 584:596 FZD receptors 584:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 49 theme cysteine-rich 554:566 arg1 CRD 576:578 CRD 576:578 CRD 576:578 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 49 theme cysteine-rich 554:566 arg1 domain 568:573 the cysteine-rich domain 550:573 the cysteine-rich domain (CRD) of FZD receptors 550:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 4 50 theme crystal 637:643 arg1 structure 645:653 a crystal structure 635:653 a crystal structure of human FZD7 CRD unexpectedly bound to a 24-carbon fatty acid 635:716 Here, we determined a crystal structure of human FZD7 CRD unexpectedly bound to a 24-carbon fatty acid. 28377511 11 51 theme structural 1510:1519 arg1 insights 1521:1528 structural insights 1510:1528 structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function 1510:1630 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 9 52 theme molecular 1166:1174 arg1 mechanism 1176:1184 a common molecular mechanism 1157:1184 a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors 1157:1319 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 5 53 theme C16:1 781:785 arg1 acid 812:815 C16:1 cis-Δ9 unsaturated fatty acid 781:815 C16:1 cis-Δ9 unsaturated fatty acid 781:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 28377511 5 54 theme unsaturated 794:804 arg1 acid 812:815 C16:1 cis-Δ9 unsaturated fatty acid 781:815 C16:1 cis-Δ9 unsaturated fatty acid 781:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 28377511 9 55 theme cis-unsaturated 1209:1223 arg1 modification 1273:1284 a necessary posttranslational modification 1243:1284 a necessary posttranslational modification of Wnts 1243:1292 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 55 theme cis-unsaturated 1209:1223 arg1 group 1236:1240 the cis-unsaturated fatty acyl group 1205:1240 the cis-unsaturated fatty acyl group 1205:1240 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 56 theme acyl 1231:1234 arg1 modification 1273:1284 a necessary posttranslational modification 1243:1284 a necessary posttranslational modification of Wnts 1243:1292 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 56 theme acyl 1231:1234 arg1 group 1236:1240 the cis-unsaturated fatty acyl group 1205:1240 the cis-unsaturated fatty acyl group 1205:1240 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 3 57 theme FZD 584:586 arg1 receptors 588:596 FZD receptors 584:596 FZD receptors 584:596 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 1 58 theme Wnt 137:139 arg1 signaling 141:149 Wnt signaling 137:149 Wnt signaling 137:149 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 0 59 theme ligand 88:93 arg1 binding 95:101 Wnt ligand binding 84:101 Wnt ligand binding 84:101 Unsaturated fatty acyl recognition by Frizzled receptors mediates dimerization upon Wnt ligand binding. 28377511 4 60 theme FZD7 664:667 arg1 CRD 669:671 human FZD7 CRD 658:671 human FZD7 CRD 658:671 Here, we determined a crystal structure of human FZD7 CRD unexpectedly bound to a 24-carbon fatty acid. 28377511 9 61 theme necessary 1245:1253 arg1 modification 1273:1284 a necessary posttranslational modification 1243:1284 a necessary posttranslational modification of Wnts 1243:1292 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 61 theme necessary 1245:1253 arg1 group 1236:1240 the cis-unsaturated fatty acyl group 1205:1240 the cis-unsaturated fatty acyl group 1205:1240 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 11 62 theme strategies 1587:1596 arg1 identification 1553:1566 the identification 1549:1566 the identification of pharmacological strategies to modulate FZD receptor function 1549:1630 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 10 63 theme Wnt 1377:1379 arg1 binding 1381:1387 Wnt binding 1377:1387 Wnt binding 1377:1387 The fatty acid bridges two CRD monomers, implying that Wnt binding mediates FZD receptor dimerization. 28377511 0 64 theme Unsaturated 0:10 arg1 recognition 23:33 Unsaturated fatty acyl recognition 0:33 Unsaturated fatty acyl recognition by Frizzled receptors 0:55 Unsaturated fatty acyl recognition by Frizzled receptors mediates dimerization upon Wnt ligand binding. 28377511 0 65 theme fatty 12:16 arg1 recognition 23:33 Unsaturated fatty acyl recognition 0:33 Unsaturated fatty acyl recognition by Frizzled receptors 0:55 Unsaturated fatty acyl recognition by Frizzled receptors mediates dimerization upon Wnt ligand binding. 28377511 9 66 theme Wnts 1289:1292 arg1 modification 1273:1284 a necessary posttranslational modification 1243:1284 a necessary posttranslational modification of Wnts 1243:1292 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 9 66 theme Wnts 1289:1292 arg1 group 1236:1240 the cis-unsaturated fatty acyl group 1205:1240 the cis-unsaturated fatty acyl group 1205:1240 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 8 67 theme published 1030:1038 arg1 structure 1055:1063 the published mouse FZD8 CRD structure 1026:1063 the published mouse FZD8 CRD structure 1026:1063 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 10 68 theme receptor 1402:1409 arg1 dimerization 1411:1422 FZD receptor dimerization 1398:1422 FZD receptor dimerization 1398:1422 The fatty acid bridges two CRD monomers, implying that Wnt binding mediates FZD receptor dimerization. 28377511 4 69 theme 24-carbon 697:705 arg1 acid 713:716 a 24-carbon fatty acid 695:716 a 24-carbon fatty acid 695:716 Here, we determined a crystal structure of human FZD7 CRD unexpectedly bound to a 24-carbon fatty acid. 28377511 2 70 theme gastric 354:360 arg1 tumors 378:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 8 71 theme FZD8 1046:1049 arg1 structure 1055:1063 the published mouse FZD8 CRD structure 1026:1063 the published mouse FZD8 CRD structure 1026:1063 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 7 72 theme lipid-binding 879:891 arg1 groove 893:898 The lipid-binding groove 875:898 The lipid-binding groove 875:898 The lipid-binding groove exhibits flexibility and spans both monomers, adopting a U-shaped geometry that accommodates the fatty acid. 28377511 1 73 theme Frizzled 104:111 arg1 receptors 119:127 Frizzled (FZD) receptors 104:127 Frizzled (FZD) receptors 104:127 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 1 73 theme Frizzled 104:111 arg1 FZD 114:116 FZD 114:116 FZD 114:116 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 3 74 theme interactions 456:467 arg1 understanding 422:434 biochemical understanding 410:434 biochemical understanding of Wnt-FZD receptor interactions 410:467 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 5 75 theme crystal 736:742 arg1 structure 744:752 a crystal structure 734:752 a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid 734:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 28377511 9 76 theme FZD 1307:1309 arg1 receptors 1311:1319 multiple FZD receptors 1298:1319 multiple FZD receptors 1298:1319 Our results define a common molecular mechanism for recognition of the cis-unsaturated fatty acyl group, a necessary posttranslational modification of Wnts, by multiple FZD receptors. 28377511 8 77 theme structure 1055:1063 arg1 Re-evaluation 1009:1021 Re-evaluation 1009:1021 Re-evaluation of the published mouse FZD8 CRD structure 1009:1063 Re-evaluation of the published mouse FZD8 CRD structure reveals that it also shares the same architecture as FZD5 and FZD7 CRDs. 28377511 1 78 theme stem 200:203 arg1 activity 210:217 stem cell activity 200:217 stem cell activity 200:217 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 3 79 theme biochemical 410:420 arg1 understanding 422:434 biochemical understanding 410:434 biochemical understanding of Wnt-FZD receptor interactions 410:467 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 3 80 theme Wnt-FZD 439:445 arg1 interactions 456:467 Wnt-FZD receptor interactions 439:467 Wnt-FZD receptor interactions 439:467 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 2 81 theme ovarian 345:351 arg1 tumors 378:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 pancreatic, ovarian, gastric, and colorectal tumors 333:383 Moreover, high FZD receptor expression at the cell surface contributes to overactive Wnt signaling in subsets of pancreatic, ovarian, gastric, and colorectal tumors. 28377511 10 82 theme fatty 1326:1330 arg1 acid 1332:1335 The fatty acid 1322:1335 The fatty acid 1322:1335 The fatty acid bridges two CRD monomers, implying that Wnt binding mediates FZD receptor dimerization. 28377511 3 83 theme Wnt 509:511 arg1 groups 540:545 Wnt cis-unsaturated fatty acyl groups 509:545 Wnt cis-unsaturated fatty acyl groups 509:545 Despite the progress in biochemical understanding of Wnt-FZD receptor interactions, the molecular basis for recognition of Wnt cis-unsaturated fatty acyl groups by the cysteine-rich domain (CRD) of FZD receptors remains elusive. 28377511 1 84 theme cell 205:208 arg1 activity 210:217 stem cell activity 200:217 stem cell activity 200:217 Frizzled (FZD) receptors mediate Wnt signaling in diverse processes ranging from bone growth to stem cell activity. 28377511 11 85 theme receptor 1614:1621 arg1 function 1623:1630 FZD receptor function 1610:1630 FZD receptor function 1610:1630 Our data uncover possibilities for the arrangement of Wnt-FZD CRD complexes and shed structural insights that could aide in the identification of pharmacological strategies to modulate FZD receptor function. 28377511 5 86 theme FZD5 763:766 arg1 CRD 768:770 human FZD5 CRD 757:770 human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid 757:815 We also report a crystal structure of human FZD5 CRD bound to C16:1 cis-Δ9 unsaturated fatty acid. 30069874 6 0 theme LYPD6 822:826 arg1 motif 832:836 the LYPD6 NxI motif 818:836 the LYPD6 NxI motif 818:836 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 5 1 theme family 754:759 arg1 members 761:767 the Ly6 protein family members 738:767 the Ly6 protein family members 738:767 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 3 2 theme LU 460:461 arg1 domain 463:468 the LYPD6 extracellular LU domain 436:468 the LYPD6 extracellular LU domain 436:468 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 6 3 theme LYPD6 944:948 arg1 interaction 929:939 the interaction 925:939 the interaction of LYPD6 with LRP6 925:958 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 3 4 theme domain 463:468 arg1 structure 423:431 the 1.25 Å resolution structure 401:431 the 1.25 Å resolution structure of the LYPD6 extracellular LU domain 401:468 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 0 5 theme coreceptor 80:89 arg1 LRP6 91:94 the Wnt coreceptor LRP6 72:94 the Wnt coreceptor LRP6 72:94 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 5 6 contain has 781:783 arg2 motif 792:796 an NxI motif 785:796 an NxI motif 785:796 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 5 6 contain has 781:783 arg1 LYPD6 775:779 only LYPD6 770:779 only LYPD6 770:779 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 6 7 with interaction 865:875 arg1 LRP6 882:885 LRP6 882:885 LRP6 882:885 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 4 8 theme tripeptide 668:677 arg1 fold 611:614 a 'trifingered protein domain' fold 580:614 a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif 580:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 8 theme tripeptide 668:677 arg1 motif 679:683 a tripeptide motif 666:683 a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors 666:732 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 0 9 theme Wnt 76:78 arg1 LRP6 91:94 the Wnt coreceptor LRP6 72:94 the Wnt coreceptor LRP6 72:94 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 1 10 dep lipoprotein 279:289 arg1 protein 308:314 receptor-related protein 6 291:316 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 10 dep lipoprotein 279:289 arg1 LRP6 319:322 LRP6 319:322 LRP6 319:322 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 6 11 from role 917:920 arg1 interaction 929:939 the interaction 925:939 the interaction of LYPD6 with LRP6 925:958 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 0 12 with interactions 54:65 arg1 LRP6 91:94 the Wnt coreceptor LRP6 72:94 the Wnt coreceptor LRP6 72:94 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 6 13 from mutations 805:813 arg1 motif 832:836 the LYPD6 NxI motif 818:836 the LYPD6 NxI motif 818:836 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 3 14 theme LYPD6 440:444 arg1 domain 463:468 the LYPD6 extracellular LU domain 436:468 the LYPD6 extracellular LU domain 436:468 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 2 15 theme acetylcholine 354:366 arg1 nAChR 378:382 nAChR 378:382 nAChR 378:382 It also binds the nicotinic acetylcholine receptor (nAChR). 30069874 2 15 theme acetylcholine 354:366 arg1 receptor 368:375 the nicotinic acetylcholine receptor 340:375 the nicotinic acetylcholine receptor (nAChR) 340:383 It also binds the nicotinic acetylcholine receptor (nAChR). 30069874 4 16 theme LRP5/6 701:706 arg1 binding 708:714 LRP5/6 binding 701:714 LRP5/6 binding by Wnt inhibitors 701:732 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 2 17 theme nicotinic 344:352 arg1 nAChR 378:382 nAChR 378:382 nAChR 378:382 It also binds the nicotinic acetylcholine receptor (nAChR). 30069874 2 17 theme nicotinic 344:352 arg1 receptor 368:375 the nicotinic acetylcholine receptor 340:375 the nicotinic acetylcholine receptor (nAChR) 340:383 It also binds the nicotinic acetylcholine receptor (nAChR). 30069874 3 18 theme surface 523:529 arg1 resonance 539:547 surface plasmon resonance 523:547 surface plasmon resonance 523:547 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 5 19 theme NxI 788:790 arg1 motif 792:796 an NxI motif 785:796 an NxI motif 785:796 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 0 20 theme Wnt 17:19 arg1 LYPD6 40:44 the Wnt signaling enhancer LYPD6 13:44 the Wnt signaling enhancer LYPD6 13:44 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 3 21 theme resolution 412:421 arg1 structure 423:431 the 1.25 Å resolution structure 401:431 the 1.25 Å resolution structure of the LYPD6 extracellular LU domain 401:468 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 5 22 theme protein 746:752 arg1 members 761:767 the Ly6 protein family members 738:767 the Ly6 protein family members 738:767 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 1 23 theme Wnt 252:254 arg1 lipoprotein 279:289 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 6 24 theme key 913:915 arg1 role 917:920 its key role 909:920 its key role in the interaction of LYPD6 with LRP6 909:958 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 1 25 contain containing 166:175 arg1 domain 159:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain 97:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 25 contain containing 166:175 arg2 LYPD6 180:184 LYPD6 180:184 LYPD6 180:184 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 25 contain containing 166:175 arg2 6 177:177 6 177:177 6 177:177 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 25 contain containing 166:175 arg1 enhancer 206:213 a Wnt signaling enhancer 190:213 a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 190:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 3 26 theme extracellular 446:458 arg1 domain 463:468 the LYPD6 extracellular LU domain 436:468 the LYPD6 extracellular LU domain 436:468 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 0 27 theme enhancer 31:38 arg1 LYPD6 40:44 the Wnt signaling enhancer LYPD6 13:44 the Wnt signaling enhancer LYPD6 13:44 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 1 28 theme coreceptor 256:265 arg1 lipoprotein 279:289 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 4 29 theme domain 603:608 arg1 fold 611:614 a 'trifingered protein domain' fold 580:614 a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif 580:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 29 theme domain 603:608 arg1 motif 679:683 a tripeptide motif 666:683 a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors 666:732 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 0 30 theme signaling 21:29 arg1 LYPD6 40:44 the Wnt signaling enhancer LYPD6 13:44 the Wnt signaling enhancer LYPD6 13:44 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 1 31 theme low 267:269 arg1 lipoprotein 279:289 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 3 32 with interaction 482:492 arg1 LRP6 499:502 LRP6 499:502 LRP6 499:502 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 4 33 with fold 611:614 arg1 fingertip 632:640 the middle fingertip 621:640 the middle fingertip bearing an 'NxI' motif 621:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 5 34 theme Ly6 742:744 arg1 members 761:767 the Ly6 protein family members 738:767 the Ly6 protein family members 738:767 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 5 35 theme only 770:773 arg1 LYPD6 775:779 only LYPD6 770:779 only LYPD6 770:779 Of the Ly6 protein family members, only LYPD6 has an NxI motif. 30069874 3 36 theme 1.25 Å 405:410 arg1 structure 423:431 the 1.25 Å resolution structure 401:431 the 1.25 Å resolution structure of the LYPD6 extracellular LU domain 401:468 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 1 37 theme density 271:277 arg1 lipoprotein 279:289 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 3 38 theme plasmon 531:537 arg1 resonance 539:547 surface plasmon resonance 523:547 surface plasmon resonance 523:547 We report here the 1.25 Å resolution structure of the LYPD6 extracellular LU domain and map its interaction with LRP6 by mutagenesis and surface plasmon resonance. 30069874 1 39 theme Ly6/urokinase-type 97:114 arg1 uPAR 148:151 uPAR 148:151 uPAR 148:151 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 39 theme Ly6/urokinase-type 97:114 arg1 LU 155:156 LU 155:156 LU 155:156 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 39 theme Ly6/urokinase-type 97:114 arg1 receptor 138:145 Ly6/urokinase-type plasminogen activator receptor 97:145 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 0 40 theme LYPD6 40:44 arg1 Structure 0:8 Structure 0:8 Structure of the Wnt signaling enhancer LYPD6 0:44 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 0 40 theme LYPD6 40:44 arg1 interactions 54:65 its interactions 50:65 its interactions with the Wnt coreceptor LRP6 50:94 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 1 41 theme lipoprotein 279:289 arg1 phosphorylation 229:243 phosphorylation 229:243 phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 229:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 6 42 with interaction 929:939 arg1 LRP6 955:958 LRP6 955:958 LRP6 955:958 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 1 43 theme plasminogen 116:126 arg1 uPAR 148:151 uPAR 148:151 uPAR 148:151 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 43 theme plasminogen 116:126 arg1 LU 155:156 LU 155:156 LU 155:156 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 43 theme plasminogen 116:126 arg1 receptor 138:145 Ly6/urokinase-type plasminogen activator receptor 97:145 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 44 theme receptor-related 291:306 arg1 protein 308:314 receptor-related protein 6 291:316 the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 248:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 44 theme receptor-related 291:306 arg1 LRP6 319:322 LRP6 319:322 LRP6 319:322 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 4 45 theme NxI 654:656 arg1 motif 659:663 an 'NxI' motif 650:663 an 'NxI' motif 650:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 46 theme Wnt 719:721 arg1 inhibitors 723:732 Wnt inhibitors 719:732 Wnt inhibitors 719:732 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 0 47 with Structure 0:8 arg1 LRP6 91:94 the Wnt coreceptor LRP6 72:94 the Wnt coreceptor LRP6 72:94 Structure of the Wnt signaling enhancer LYPD6 and its interactions with the Wnt coreceptor LRP6. 30069874 1 48 theme activator 128:136 arg1 uPAR 148:151 uPAR 148:151 uPAR 148:151 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 48 theme activator 128:136 arg1 LU 155:156 LU 155:156 LU 155:156 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 48 theme activator 128:136 arg1 receptor 138:145 Ly6/urokinase-type plasminogen activator receptor 97:145 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 49 theme receptor 138:145 arg1 domain 159:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain 97:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 49 theme receptor 138:145 arg1 enhancer 206:213 a Wnt signaling enhancer 190:213 a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 190:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 4 50 theme LYPD6LU 554:560 arg1 structure 562:570 The LYPD6LU structure 550:570 The LYPD6LU structure 550:570 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 51 theme protein 595:601 arg1 fold 611:614 a 'trifingered protein domain' fold 580:614 a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif 580:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 51 theme protein 595:601 arg1 motif 679:683 a tripeptide motif 666:683 a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors 666:732 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 52 theme trifingered 583:593 arg1 fold 611:614 a 'trifingered protein domain' fold 580:614 a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif 580:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 4 52 theme trifingered 583:593 arg1 motif 679:683 a tripeptide motif 666:683 a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors 666:732 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 1 53 theme Wnt 192:194 arg1 domain 159:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain 97:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 53 theme Wnt 192:194 arg1 enhancer 206:213 a Wnt signaling enhancer 190:213 a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 190:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 6 54 theme NxI 828:830 arg1 motif 832:836 the LYPD6 NxI motif 818:836 the LYPD6 NxI motif 818:836 Since mutations in the LYPD6 NxI motif abolish or severely reduce interaction with LRP6, our results indicate its key role in the interaction of LYPD6 with LRP6. 30069874 4 55 theme middle 625:630 arg1 fingertip 632:640 the middle fingertip 621:640 the middle fingertip bearing an 'NxI' motif 621:663 The LYPD6LU structure reveals a 'trifingered protein domain' fold with the middle fingertip bearing an 'NxI' motif, a tripeptide motif associated with LRP5/6 binding by Wnt inhibitors. 30069874 1 56 theme signaling 196:204 arg1 domain 159:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain 97:164 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) 97:185 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 30069874 1 56 theme signaling 196:204 arg1 enhancer 206:213 a Wnt signaling enhancer 190:213 a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6) 190:323 Ly6/urokinase-type plasminogen activator receptor (uPAR) (LU) domain containing 6 (LYPD6) is a Wnt signaling enhancer that promotes phosphorylation of the Wnt coreceptor low density lipoprotein receptor-related protein 6 (LRP6). 23260142 7 0 theme selection 910:918 arg1 process 920:926 a rapid and stringent selection process 888:926 a rapid and stringent selection process for the highest-affinity binders 888:959 This energetic barrier creates a rapid and stringent selection process for the highest-affinity binders. 23260142 6 1 theme HLA-DR 784:789 arg1 residues 791:798 HLA-DR residues 784:798 HLA-DR residues 784:798 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 0 2 theme peptide 77:83 arg1 selection 85:93 rapid peptide selection 71:93 rapid peptide selection 71:93 Crystal structure of the HLA-DM-HLA-DR1 complex defines mechanisms for rapid peptide selection. 23260142 6 3 theme hydrophobic 834:844 arg1 pocket 849:854 the hydrophobic P1 pocket 830:854 the hydrophobic P1 pocket 830:854 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 7 4 theme rapid 890:894 arg1 process 920:926 a rapid and stringent selection process 888:926 a rapid and stringent selection process for the highest-affinity binders 888:959 This energetic barrier creates a rapid and stringent selection process for the highest-affinity binders. 23260142 4 5 theme HLA-DR 532:537 arg1 residues 539:546 hydrophobic HLA-DR residues 520:546 hydrophobic HLA-DR residues 520:546 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 8 6 theme P2 1001:1002 arg1 sites 1011:1015 the P2 and P1 sites 997:1015 the P2 and P1 sites 997:1015 Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation. 23260142 2 7 theme selection 285:293 arg1 process 295:301 the endosomal peptide selection process 263:301 the endosomal peptide selection process 263:301 HLA-DM plays a critical role in the endosomal peptide selection process. 23260142 4 8 theme conformational 483:496 arg1 changes 498:504 major conformational changes 477:504 major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket 477:565 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 5 9 theme empty 647:651 arg1 groove 676:681 the empty HLA-DR peptide-binding groove 643:681 the empty HLA-DR peptide-binding groove 643:681 These conformational changes accelerate peptide dissociation and stabilize the empty HLA-DR peptide-binding groove. 23260142 2 10 theme peptide 277:283 arg1 process 295:301 the endosomal peptide selection process 263:301 the endosomal peptide selection process 263:301 HLA-DM plays a critical role in the endosomal peptide selection process. 23260142 6 11 theme groove 752:757 arg1 part 733:736 only part 728:736 only part of the HLA-DR groove 728:757 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 4 12 theme major 477:481 arg1 changes 498:504 major conformational changes 477:504 major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket 477:565 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 5 13 theme peptide-binding 660:674 arg1 groove 676:681 the empty HLA-DR peptide-binding groove 643:681 the empty HLA-DR peptide-binding groove 643:681 These conformational changes accelerate peptide dissociation and stabilize the empty HLA-DR peptide-binding groove. 23260142 5 14 theme HLA-DR 653:658 arg1 groove 676:681 the empty HLA-DR peptide-binding groove 643:681 the empty HLA-DR peptide-binding groove 643:681 These conformational changes accelerate peptide dissociation and stabilize the empty HLA-DR peptide-binding groove. 23260142 2 15 theme endosomal 267:275 arg1 process 295:301 the endosomal peptide selection process 263:301 the endosomal peptide selection process 263:301 HLA-DM plays a critical role in the endosomal peptide selection process. 23260142 6 16 theme HLA-DR 745:750 arg1 groove 752:757 the HLA-DR groove 741:757 the HLA-DR groove 741:757 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 3 17 theme major 353:357 arg1 rearrangements 359:372 major rearrangements 353:372 major rearrangements of the HLA-DR peptide-binding groove 353:409 The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. 23260142 1 18 attach present 169:175 arg2 molecules 103:111 HLA-DR molecules 96:111 HLA-DR molecules 96:111 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 1 18 attach present 169:175 arg1 surface 194:200 the cell surface 185:200 the cell surface 185:200 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 8 19 theme DM 1084:1085 arg1 dissociation 1087:1098 DM dissociation 1084:1098 DM dissociation 1084:1098 Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation. 23260142 6 20 theme P2 818:819 arg1 site 821:824 the P2 site 814:824 the P2 site 814:824 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 0 21 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the HLA-DM-HLA-DR1 complex 0:46 Crystal structure of the HLA-DM-HLA-DR1 complex defines mechanisms for rapid peptide selection. 23260142 7 22 theme energetic 862:870 arg1 barrier 872:878 This energetic barrier 857:878 This energetic barrier 857:878 This energetic barrier creates a rapid and stringent selection process for the highest-affinity binders. 23260142 3 23 theme peptide-binding 388:402 arg1 groove 404:409 the HLA-DR peptide-binding groove 377:409 the HLA-DR peptide-binding groove 377:409 The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. 23260142 0 24 theme rapid 71:75 arg1 selection 85:93 rapid peptide selection 71:93 rapid peptide selection 71:93 Crystal structure of the HLA-DM-HLA-DR1 complex defines mechanisms for rapid peptide selection. 23260142 3 25 theme groove 404:409 arg1 rearrangements 359:372 major rearrangements 353:372 major rearrangements of the HLA-DR peptide-binding groove 353:409 The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. 23260142 1 26 theme HLA-DR 96:101 arg1 molecules 103:111 HLA-DR molecules 96:111 HLA-DR molecules 96:111 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 1 27 theme cell 189:192 arg1 surface 194:200 the cell surface 185:200 the cell surface 185:200 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 1 28 from peptides 128:135 arg1 compartment 153:163 an endosomal compartment 140:163 an endosomal compartment 140:163 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 3 29 theme HLA-DR 381:386 arg1 groove 404:409 the HLA-DR peptide-binding groove 377:409 the HLA-DR peptide-binding groove 377:409 The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. 23260142 4 30 from pocket 462:467 arg1 Flipping 412:419 Flipping 412:419 Flipping of a tryptophan away from the HLA-DR1 P1 pocket 412:467 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 4 31 theme P1 459:460 arg1 pocket 462:467 the HLA-DR1 P1 pocket 447:467 the HLA-DR1 P1 pocket 447:467 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 0 32 theme complex 40:46 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the HLA-DM-HLA-DR1 complex 0:46 Crystal structure of the HLA-DM-HLA-DR1 complex defines mechanisms for rapid peptide selection. 23260142 4 33 theme HLA-DR1 451:457 arg1 pocket 462:467 the HLA-DR1 P1 pocket 447:467 the HLA-DR1 P1 pocket 447:467 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 1 34 theme CD4 206:208 arg1 surveillance 217:228 CD4 T cell surveillance 206:228 CD4 T cell surveillance 206:228 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 0 35 theme HLA-DM-HLA-DR1 25:38 arg1 complex 40:46 the HLA-DM-HLA-DR1 complex 21:46 the HLA-DM-HLA-DR1 complex 21:46 Crystal structure of the HLA-DM-HLA-DR1 complex defines mechanisms for rapid peptide selection. 23260142 6 36 contain have 713:716 arg2 access 718:723 access 718:723 access to only part of the HLA-DR groove 718:757 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 6 36 contain have 713:716 arg1 peptides 704:711 incoming peptides 695:711 incoming peptides 695:711 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 8 37 theme conformational 1030:1043 arg1 changes 1045:1051 the conformational changes 1026:1051 the conformational changes 1026:1051 Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation. 23260142 1 38 theme T cell 210:215 arg1 surveillance 217:228 CD4 T cell surveillance 206:228 CD4 T cell surveillance 206:228 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 1 39 theme microbial 118:126 arg1 peptides 128:135 microbial peptides 118:135 microbial peptides in an endosomal compartment 118:163 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 5 40 theme conformational 574:587 arg1 changes 589:595 These conformational changes 568:595 These conformational changes 568:595 These conformational changes accelerate peptide dissociation and stabilize the empty HLA-DR peptide-binding groove. 23260142 4 41 theme P1 557:558 arg1 pocket 560:565 the P1 pocket 553:565 the P1 pocket 553:565 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 4 42 theme tryptophan 426:435 arg1 Flipping 412:419 Flipping 412:419 Flipping of a tryptophan away from the HLA-DR1 P1 pocket 412:467 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 8 43 theme P1 1008:1009 arg1 sites 1011:1015 the P2 and P1 sites 997:1015 the P2 and P1 sites 997:1015 Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation. 23260142 5 44 theme peptide 608:614 arg1 dissociation 616:627 peptide dissociation 608:627 peptide dissociation 608:627 These conformational changes accelerate peptide dissociation and stabilize the empty HLA-DR peptide-binding groove. 23260142 8 45 theme residues 983:990 arg1 Insertion 962:970 Insertion 962:970 Insertion of peptide residues into the P2 and P1 sites 962:1015 Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation. 23260142 6 46 theme incoming 695:702 arg1 peptides 704:711 incoming peptides 695:711 incoming peptides 695:711 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 3 47 theme HLA-DM-HLA-DR 325:337 arg1 complex 339:345 the HLA-DM-HLA-DR complex 321:345 the HLA-DM-HLA-DR complex 321:345 The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. 23260142 6 48 theme P1 846:847 arg1 pocket 849:854 the hydrophobic P1 pocket 830:854 the hydrophobic P1 pocket 830:854 Initially, incoming peptides have access to only part of the HLA-DR groove and need to compete with HLA-DR residues for access to the P2 site and the hydrophobic P1 pocket. 23260142 2 49 theme critical 246:253 arg1 role 255:258 a critical role 244:258 a critical role 244:258 HLA-DM plays a critical role in the endosomal peptide selection process. 23260142 8 50 theme peptide 975:981 arg1 residues 983:990 peptide residues 975:990 peptide residues 975:990 Insertion of peptide residues into the P2 and P1 sites reverses the conformational changes, terminating selection through DM dissociation. 23260142 1 51 theme endosomal 143:151 arg1 compartment 153:163 an endosomal compartment 140:163 an endosomal compartment 140:163 HLA-DR molecules bind microbial peptides in an endosomal compartment and present them on the cell surface for CD4 T cell surveillance. 23260142 7 52 theme highest-affinity 936:951 arg1 binders 953:959 the highest-affinity binders 932:959 the highest-affinity binders 932:959 This energetic barrier creates a rapid and stringent selection process for the highest-affinity binders. 23260142 3 53 theme complex 339:345 arg1 structure 308:316 The structure 304:316 The structure of the HLA-DM-HLA-DR complex 304:345 The structure of the HLA-DM-HLA-DR complex shows major rearrangements of the HLA-DR peptide-binding groove. 23260142 4 54 theme hydrophobic 520:530 arg1 residues 539:546 hydrophobic HLA-DR residues 520:546 hydrophobic HLA-DR residues 520:546 Flipping of a tryptophan away from the HLA-DR1 P1 pocket enables major conformational changes that position hydrophobic HLA-DR residues into the P1 pocket. 23260142 7 55 theme stringent 900:908 arg1 process 920:926 a rapid and stringent selection process 888:926 a rapid and stringent selection process for the highest-affinity binders 888:959 This energetic barrier creates a rapid and stringent selection process for the highest-affinity binders. 20505120 6 0 theme interfaces 930:939 arg1 importance 885:894 the importance 881:894 the importance of the identified oligomerization interfaces for Eph clustering 881:958 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 4 1 with complexes 497:505 arg1 A5 526:527 A5 526:527 A5 526:527 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 1 with complexes 497:505 arg1 ephrin-A1 512:520 ephrin-A1 512:520 ephrin-A1 512:520 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 2 theme Eph 552:554 arg1 cluster 556:562 an Eph cluster 549:562 an Eph cluster 549:562 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 6 3 theme oligomerization 914:928 arg1 interfaces 930:939 the identified oligomerization interfaces 899:939 the identified oligomerization interfaces 899:939 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 3 4 theme mechanistic 360:370 arg1 details 372:378 the structural and mechanistic details 341:378 the structural and mechanistic details of this assembly 341:395 However, the structural and mechanistic details of this assembly remained undefined. 20505120 6 5 theme signaling 854:862 arg1 assays 864:869 cell-based signaling assays 843:869 cell-based signaling assays 843:869 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 2 6 theme bidirectional 286:298 arg1 signaling 300:308 bidirectional signaling 286:308 bidirectional signaling 286:308 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 5 7 theme novel 618:622 arg1 interactions 632:643 novel Eph/Eph interactions 618:643 novel Eph/Eph interactions 618:643 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 3 8 theme structural 345:354 arg1 details 372:378 the structural and mechanistic details 341:378 the structural and mechanistic details of this assembly 341:395 However, the structural and mechanistic details of this assembly remained undefined. 20505120 6 9 theme Eph 945:947 arg1 clustering 949:958 Eph clustering 945:958 Eph clustering 945:958 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 5 10 theme Eph/Eph 624:630 arg1 interactions 632:643 novel Eph/Eph interactions 618:643 novel Eph/Eph interactions 618:643 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 5 11 theme Eph/ephrin 752:761 arg1 clustering 763:772 Eph/ephrin clustering 752:772 Eph/ephrin clustering 752:772 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 3 12 theme assembly 388:395 arg1 details 372:378 the structural and mechanistic details 341:378 the structural and mechanistic details of this assembly 341:395 However, the structural and mechanistic details of this assembly remained undefined. 20505120 1 13 theme tyrosine 52:59 arg1 kinases 61:67 Eph receptor tyrosine kinases 39:67 Eph receptor tyrosine kinases 39:67 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20505120 6 14 theme site-directed 813:825 arg1 mutagenesis 827:837 site-directed mutagenesis 813:837 site-directed mutagenesis 813:837 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 1 15 theme ephrin 79:84 arg1 ligands 86:92 their ephrin ligands 73:92 their ephrin ligands 73:92 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20505120 4 16 theme cluster 556:562 arg1 structures 449:458 high-resolution structures 433:458 high-resolution structures of the complete EphA2 ectodomain 433:491 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 16 theme cluster 556:562 arg1 unit 541:544 the base unit 532:544 the base unit of an Eph cluster 532:562 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 16 theme cluster 556:562 arg1 complexes 497:505 complexes 497:505 complexes with ephrin-A1 and A5 497:527 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 2 17 theme interacting 313:323 arg1 cells 325:329 interacting cells 313:329 interacting cells 313:329 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 0 18 theme receptor 20:27 arg1 clusters 29:36 Eph receptor clusters 16:36 Eph receptor clusters 16:36 Architecture of Eph receptor clusters. 20505120 2 19 theme higher-order 241:252 arg1 clusters 264:271 higher-order signaling clusters 241:271 higher-order signaling clusters that set off bidirectional signaling in interacting cells 241:329 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 5 20 theme elongated 590:598 arg1 architecture 600:611 an elongated architecture 587:611 an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering 587:772 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 0 21 theme Eph 16:18 arg1 clusters 29:36 Eph receptor clusters 16:36 Eph receptor clusters 16:36 Architecture of Eph receptor clusters. 20505120 4 22 theme complete 467:474 arg1 ectodomain 482:491 the complete EphA2 ectodomain 463:491 the complete EphA2 ectodomain 463:491 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 1 23 theme cell 103:106 arg1 navigation 108:117 cell navigation 103:117 cell navigation 103:117 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20505120 6 24 theme cell-based 843:852 arg1 assays 864:869 cell-based signaling assays 843:869 cell-based signaling assays 843:869 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 4 25 with structures 449:458 arg1 A5 526:527 A5 526:527 A5 526:527 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 25 with structures 449:458 arg1 ephrin-A1 512:520 ephrin-A1 512:520 ephrin-A1 512:520 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 5 26 dep both 646:649 arg1 within 651:656 within 651:656 within 651:656 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 6 27 theme identified 903:912 arg1 interfaces 930:939 the identified oligomerization interfaces 899:939 the identified oligomerization interfaces 899:939 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 0 28 theme clusters 29:36 arg1 Architecture 0:11 Architecture 0:11 Architecture of Eph receptor clusters. 0:37 Architecture of Eph receptor clusters. 20505120 5 29 theme molecular 721:729 arg1 mechanism 731:739 the molecular mechanism 717:739 the molecular mechanism underlying Eph/ephrin clustering 717:772 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 2 30 theme Ephs 173:176 arg1 Signaling 160:168 Signaling 160:168 Signaling of Ephs 160:176 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 5 31 dep interactions 632:643 arg1 both 646:649 both 646:649 both 646:649 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 4 32 theme base 536:539 arg1 structures 449:458 high-resolution structures 433:458 high-resolution structures of the complete EphA2 ectodomain 433:491 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 32 theme base 536:539 arg1 unit 541:544 the base unit 532:544 the base unit of an Eph cluster 532:562 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 32 theme base 536:539 arg1 complexes 497:505 complexes 497:505 complexes with ephrin-A1 and A5 497:527 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 5 33 theme Eph 677:679 arg1 domain 696:701 the Eph ligand-binding domain 673:701 the Eph ligand-binding domain 673:701 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 4 34 theme EphA2 476:480 arg1 ectodomain 482:491 the complete EphA2 ectodomain 463:491 the complete EphA2 ectodomain 463:491 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 5 35 with architecture 600:611 arg1 interactions 632:643 novel Eph/Eph interactions 618:643 novel Eph/Eph interactions 618:643 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 5 36 theme ligand-binding 681:694 arg1 domain 696:701 the Eph ligand-binding domain 673:701 the Eph ligand-binding domain 673:701 The structures reveal an elongated architecture with novel Eph/Eph interactions, both within and outside of the Eph ligand-binding domain, that suggest the molecular mechanism underlying Eph/ephrin clustering. 20505120 4 37 theme high-resolution 433:447 arg1 structures 449:458 high-resolution structures 433:458 high-resolution structures of the complete EphA2 ectodomain 433:491 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 37 theme high-resolution 433:447 arg1 unit 541:544 the base unit 532:544 the base unit of an Eph cluster 532:562 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 37 theme high-resolution 433:447 arg1 complexes 497:505 complexes 497:505 complexes with ephrin-A1 and A5 497:527 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 6 38 theme Structure-function 775:792 arg1 analysis 794:801 Structure-function analysis 775:801 Structure-function analysis 775:801 Structure-function analysis, by using site-directed mutagenesis and cell-based signaling assays, confirms the importance of the identified oligomerization interfaces for Eph clustering. 20505120 4 39 theme ectodomain 482:491 arg1 structures 449:458 high-resolution structures 433:458 high-resolution structures of the complete EphA2 ectodomain 433:491 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 39 theme ectodomain 482:491 arg1 complexes 497:505 complexes 497:505 complexes with ephrin-A1 and A5 497:527 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 4 39 theme ectodomain 482:491 arg1 unit 541:544 the base unit 532:544 the base unit of an Eph cluster 532:562 Here we present high-resolution structures of the complete EphA2 ectodomain and complexes with ephrin-A1 and A5 as the base unit of an Eph cluster. 20505120 2 40 theme multistep 196:204 arg1 process 206:212 a multistep process 194:212 a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells 194:329 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 2 41 theme clusters 264:271 arg1 assembly 229:236 the assembly 225:236 the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells 225:329 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 1 42 theme normal 126:131 arg1 development 147:157 normal and oncogenic development 126:157 normal and oncogenic development 126:157 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20505120 2 43 theme signaling 254:262 arg1 clusters 264:271 higher-order signaling clusters 241:271 higher-order signaling clusters that set off bidirectional signaling in interacting cells 241:329 Signaling of Ephs is initiated in a multistep process leading to the assembly of higher-order signaling clusters that set off bidirectional signaling in interacting cells. 20505120 1 44 theme Eph 39:41 arg1 kinases 61:67 Eph receptor tyrosine kinases 39:67 Eph receptor tyrosine kinases 39:67 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20505120 1 45 theme oncogenic 137:145 arg1 development 147:157 normal and oncogenic development 126:157 normal and oncogenic development 126:157 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20505120 1 46 theme receptor 43:50 arg1 kinases 61:67 Eph receptor tyrosine kinases 39:67 Eph receptor tyrosine kinases 39:67 Eph receptor tyrosine kinases and their ephrin ligands regulate cell navigation during normal and oncogenic development. 20534510 0 0 theme factor/receptor 96:110 arg1 complex 112:118 a platelet-derived growth factor/receptor complex 70:118 a platelet-derived growth factor/receptor complex 70:118 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 6 1 theme receptor 1108:1115 arg1 binding 1117:1123 receptor binding 1108:1123 receptor binding 1108:1123 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 3 2 theme conserved 539:547 arg1 mode 574:577 this conserved, hydrophobic association mode 534:577 this conserved, hydrophobic association mode 534:577 The structure of the PDGF-A/propeptide complex reveals this conserved, hydrophobic association mode. 20534510 1 3 theme critical 261:268 arg1 functions 270:278 critical functions 261:278 critical functions 261:278 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 0 4 theme growth 89:94 arg1 complex 112:118 a platelet-derived growth factor/receptor complex 70:118 a platelet-derived growth factor/receptor complex 70:118 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 0 5 link platelet-derived 16:31 arg1 complex 58:64 a platelet-derived growth factor/propeptide complex 14:64 a platelet-derived growth factor/propeptide complex 14:64 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 5 6 theme propeptides 932:942 arg1 need 924:927 the need 920:927 the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion 920:1021 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 6 7 theme L1 1146:1147 arg1 loop 1149:1152 the PDGF-B L1 loop 1135:1152 the PDGF-B L1 loop 1135:1152 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 7 8 theme PDGFRalpha 1377:1386 arg1 interaction 1388:1398 the membrane-proximal homotypic PDGFRalpha interaction 1345:1398 the membrane-proximal homotypic PDGFRalpha interaction 1345:1398 Calorimetric data also shows that the membrane-proximal homotypic PDGFRalpha interaction, albeit required for activation, contributes negatively to ligand binding. 20534510 2 9 from region 333:338 arg1 sequences 359:367 their prodomain sequences 343:367 their prodomain sequences 343:367 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 7 10 theme ligand 1459:1464 arg1 binding 1466:1472 ligand binding 1459:1472 ligand binding 1459:1472 Calorimetric data also shows that the membrane-proximal homotypic PDGFRalpha interaction, albeit required for activation, contributes negatively to ligand binding. 20534510 6 11 theme structural 1038:1047 arg1 organization 1049:1060 A large-scale structural organization 1024:1060 A large-scale structural organization 1024:1060 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 0 12 theme complex 112:118 arg1 Structures 0:9 Structures 0:9 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 0:119 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 6 13 theme PDGFRbeta 1300:1308 arg1 domain 1290:1295 the third Ig domain 1277:1295 the third Ig domain of PDGFRbeta 1277:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 6 13 theme PDGFRbeta 1300:1308 arg1 PDGFRbeta 1300:1308 PDGFRbeta 1300:1308 PDGFRbeta 1300:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 5 14 theme PDGFRbeta 774:782 arg1 interface 784:792 The PDGF-B:PDGFRbeta interface 763:792 The PDGF-B:PDGFRbeta interface 763:792 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 5 14 theme PDGFRbeta 774:782 arg1 hydrophobic 811:821 hydrophobic 811:821 hydrophobic 811:821 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 4 15 theme dimerization 744:755 arg1 seam 757:760 their dimerization seam 738:760 their dimerization seam 738:760 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 5 16 theme PDGF-B 767:772 arg1 interface 784:792 The PDGF-B:PDGFRbeta interface 763:792 The PDGF-B:PDGFRbeta interface 763:792 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 5 16 theme PDGF-B 767:772 arg1 hydrophobic 811:821 hydrophobic 811:821 hydrophobic 811:821 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 7 17 theme Calorimetric 1311:1322 arg1 data 1324:1327 Calorimetric data 1311:1327 Calorimetric data 1311:1327 Calorimetric data also shows that the membrane-proximal homotypic PDGFRalpha interaction, albeit required for activation, contributes negatively to ligand binding. 20534510 5 18 theme overlapping 867:877 arg1 positions 879:887 overlapping positions 867:887 overlapping positions on mature PDGFs 867:903 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 4 19 theme Ig 660:661 arg1 PDGFRbeta 674:682 PDGFRbeta 674:682 PDGFRbeta 674:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 19 theme Ig 660:661 arg1 domains 663:669 the first three Ig domains 644:669 the first three Ig domains of PDGFRbeta 644:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 19 theme Ig 660:661 arg1 complex 617:623 the complex 613:623 the complex between PDGF-B 613:638 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 7 20 dep contributes 1433:1443 arg1 required 1408:1415 required 1408:1415 required for activation 1408:1430 Calorimetric data also shows that the membrane-proximal homotypic PDGFRalpha interaction, albeit required for activation, contributes negatively to ligand binding. 20534510 5 21 theme mature 892:897 arg1 PDGFs 899:903 mature PDGFs 892:903 mature PDGFs 892:903 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 6 22 theme large-scale 1026:1036 arg1 organization 1049:1060 A large-scale structural organization 1024:1060 A large-scale structural organization 1024:1060 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 4 23 theme complex 617:623 arg1 PDGFRbeta 674:682 PDGFRbeta 674:682 PDGFRbeta 674:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 23 theme complex 617:623 arg1 structure 600:608 the structure 596:608 the structure of the complex between PDGF-B 596:638 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 23 theme complex 617:623 arg1 domains 663:669 the first three Ig domains 644:669 the first three Ig domains of PDGFRbeta 644:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 23 theme complex 617:623 arg1 complex 617:623 the complex 613:623 the complex between PDGF-B 613:638 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 6 24 theme PDGF-B 1139:1144 arg1 loop 1149:1152 the PDGF-B L1 loop 1135:1152 the PDGF-B L1 loop 1135:1152 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 6 25 theme form 1195:1198 arg1 structure 1173:1181 the structure 1169:1181 the structure of the free form 1169:1198 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 6 26 theme Ig 1287:1288 arg1 domain 1290:1295 the third Ig domain 1277:1295 the third Ig domain of PDGFRbeta 1277:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 6 26 theme Ig 1287:1288 arg1 PDGFRbeta 1300:1308 PDGFRbeta 1300:1308 PDGFRbeta 1300:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 0 27 theme platelet-derived 72:87 arg1 complex 112:118 a platelet-derived growth factor/receptor complex 70:118 a platelet-derived growth factor/receptor complex 70:118 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 2 28 theme conserved 323:331 arg1 region 333:338 a conserved region 321:338 a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing 321:476 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 7 29 theme membrane-proximal 1349:1365 arg1 interaction 1388:1398 the membrane-proximal homotypic PDGFRalpha interaction 1345:1398 the membrane-proximal homotypic PDGFRalpha interaction 1345:1398 Calorimetric data also shows that the membrane-proximal homotypic PDGFRalpha interaction, albeit required for activation, contributes negatively to ligand binding. 20534510 6 30 theme third 1281:1285 arg1 domain 1290:1295 the third Ig domain 1277:1295 the third Ig domain of PDGFRbeta 1277:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 6 30 theme third 1281:1285 arg1 PDGFRbeta 1300:1308 PDGFRbeta 1300:1308 PDGFRbeta 1300:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 5 31 theme important 975:983 arg1 surfaces 997:1004 functionally important hydrophobic surfaces 962:1004 functionally important hydrophobic surfaces 962:1004 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 1 32 theme prototypic 194:203 arg1 factors 212:218 prototypic growth factors 194:218 prototypic growth factors 194:218 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 32 theme prototypic 194:203 arg1 factors 145:151 Platelet-derived growth factors 121:151 Platelet-derived growth factors (PDGFs) 121:159 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 32 theme prototypic 194:203 arg1 receptors 171:179 their receptors 165:179 their receptors (PDGFRs) 165:188 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 0 33 theme platelet-derived 16:31 arg1 complex 58:64 a platelet-derived growth factor/propeptide complex 14:64 a platelet-derived growth factor/propeptide complex 14:64 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 1 34 theme Platelet-derived 121:136 arg1 PDGFs 154:158 PDGFs 154:158 PDGFs 154:158 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 34 theme Platelet-derived 121:136 arg1 factors 212:218 prototypic growth factors 194:218 prototypic growth factors 194:218 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 34 theme Platelet-derived 121:136 arg1 factors 145:151 Platelet-derived growth factors 121:151 Platelet-derived growth factors (PDGFs) 121:159 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 34 theme Platelet-derived 121:136 arg1 receptors 171:179 their receptors 165:179 their receptors (PDGFRs) 165:188 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 2 35 theme factor 447:452 arg1 domain 454:459 the mature cystine-knot growth factor domain 416:459 the mature cystine-knot growth factor domain after processing 416:476 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 6 36 theme free 1190:1193 arg1 form 1195:1198 the free form 1186:1198 the free form 1186:1198 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 1 37 theme growth 138:143 arg1 PDGFs 154:158 PDGFs 154:158 PDGFs 154:158 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 37 theme growth 138:143 arg1 factors 212:218 prototypic growth factors 194:218 prototypic growth factors 194:218 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 37 theme growth 138:143 arg1 factors 145:151 Platelet-derived growth factors 121:151 Platelet-derived growth factors (PDGFs) 121:159 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 37 theme growth 138:143 arg1 receptors 171:179 their receptors 165:179 their receptors (PDGFRs) 165:188 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 2 38 theme growth 440:445 arg1 domain 454:459 the mature cystine-knot growth factor domain 416:459 the mature cystine-knot growth factor domain after processing 416:476 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 0 39 theme factor/propeptide 40:56 arg1 complex 58:64 a platelet-derived growth factor/propeptide complex 14:64 a platelet-derived growth factor/propeptide complex 14:64 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 3 40 dep conserved 539:547 arg1 hydrophobic 550:560 hydrophobic 550:560 hydrophobic 550:560 The structure of the PDGF-A/propeptide complex reveals this conserved, hydrophobic association mode. 20534510 4 41 theme PDGFRbeta 674:682 arg1 PDGFRbeta 674:682 PDGFRbeta 674:682 PDGFRbeta 674:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 41 theme PDGFRbeta 674:682 arg1 structure 600:608 the structure 596:608 the structure of the complex between PDGF-B 596:638 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 41 theme PDGFRbeta 674:682 arg1 domains 663:669 the first three Ig domains 644:669 the first three Ig domains of PDGFRbeta 644:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 41 theme PDGFRbeta 674:682 arg1 complex 617:623 the complex 613:623 the complex between PDGF-B 613:638 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 1 42 contain have 256:259 arg1 factors 145:151 Platelet-derived growth factors 121:151 Platelet-derived growth factors (PDGFs) 121:159 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 42 contain have 256:259 arg1 factors 212:218 prototypic growth factors 194:218 prototypic growth factors 194:218 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 42 contain have 256:259 arg2 functions 270:278 critical functions 261:278 critical functions 261:278 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 42 contain have 256:259 arg1 receptors 171:179 their receptors 165:179 their receptors (PDGFRs) 165:188 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 42 contain have 256:259 arg1 kinases 242:248 receptor tyrosine kinases 224:248 receptor tyrosine kinases 224:248 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 7 43 theme homotypic 1367:1375 arg1 interaction 1388:1398 the membrane-proximal homotypic PDGFRalpha interaction 1345:1398 the membrane-proximal homotypic PDGFRalpha interaction 1345:1398 Calorimetric data also shows that the membrane-proximal homotypic PDGFRalpha interaction, albeit required for activation, contributes negatively to ligand binding. 20534510 2 44 theme cystine-knot 427:438 arg1 domain 454:459 the mature cystine-knot growth factor domain 416:459 the mature cystine-knot growth factor domain after processing 416:476 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 0 45 theme growth 33:38 arg1 complex 58:64 a platelet-derived growth factor/propeptide complex 14:64 a platelet-derived growth factor/propeptide complex 14:64 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 5 46 theme PDGF 843:846 arg1 propeptides 848:858 the PDGF propeptides 839:858 the PDGF propeptides 839:858 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 5 47 from positions 879:887 arg1 PDGFs 899:903 mature PDGFs 892:903 mature PDGFs 892:903 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 1 48 theme receptor 224:231 arg1 kinases 242:248 receptor tyrosine kinases 224:248 receptor tyrosine kinases 224:248 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 2 49 theme prodomain 349:357 arg1 sequences 359:367 their prodomain sequences 343:367 their prodomain sequences 343:367 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 0 50 theme complex 58:64 arg1 Structures 0:9 Structures 0:9 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 0:119 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 8 51 theme PDGF-PDGFR 1540:1549 arg1 signaling 1551:1559 PDGF-PDGFR signaling 1540:1559 PDGF-PDGFR signaling 1540:1559 The structural and biochemical data together offer insights into PDGF-PDGFR signaling, as well as strategies for PDGF-antagonism. 20534510 6 52 theme hydrophobic 1247:1257 arg1 interactions 1259:1270 hydrophobic interactions 1247:1270 hydrophobic interactions 1247:1270 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 0 53 link platelet-derived 72:87 arg1 complex 112:118 a platelet-derived growth factor/receptor complex 70:118 a platelet-derived growth factor/receptor complex 70:118 Structures of a platelet-derived growth factor/propeptide complex and a platelet-derived growth factor/receptor complex. 20534510 8 54 theme structural 1479:1488 arg1 data 1506:1509 The structural and biochemical data 1475:1509 The structural and biochemical data 1475:1509 The structural and biochemical data together offer insights into PDGF-PDGFR signaling, as well as strategies for PDGF-antagonism. 20534510 4 55 theme PDGF-B 702:707 arg1 protomers 709:717 two PDGF-B protomers 698:717 two PDGF-B protomers 698:717 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 3 56 theme PDGF-A/propeptide 500:516 arg1 complex 518:524 the PDGF-A/propeptide complex 496:524 the PDGF-A/propeptide complex 496:524 The structure of the PDGF-A/propeptide complex reveals this conserved, hydrophobic association mode. 20534510 5 57 theme hydrophobic 985:995 arg1 surfaces 997:1004 functionally important hydrophobic surfaces 962:1004 functionally important hydrophobic surfaces 962:1004 The PDGF-B:PDGFRbeta interface is predominantly hydrophobic, and PDGFRs and the PDGF propeptides occupy overlapping positions on mature PDGFs, rationalizing the need of propeptides by PDGFs to cover functionally important hydrophobic surfaces during secretion. 20534510 1 58 theme tyrosine 233:240 arg1 kinases 242:248 receptor tyrosine kinases 224:248 receptor tyrosine kinases 224:248 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 3 59 theme complex 518:524 arg1 structure 483:491 The structure 479:491 The structure of the PDGF-A/propeptide complex 479:524 The structure of the PDGF-A/propeptide complex reveals this conserved, hydrophobic association mode. 20534510 1 60 link Platelet-derived 121:136 arg1 PDGFs 154:158 PDGFs 154:158 PDGFs 154:158 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 60 link Platelet-derived 121:136 arg1 factors 212:218 prototypic growth factors 194:218 prototypic growth factors 194:218 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 60 link Platelet-derived 121:136 arg1 factors 145:151 Platelet-derived growth factors 121:151 Platelet-derived growth factors (PDGFs) 121:159 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 60 link Platelet-derived 121:136 arg1 receptors 171:179 their receptors 165:179 their receptors (PDGFRs) 165:188 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 2 61 theme mature 420:425 arg1 domain 454:459 the mature cystine-knot growth factor domain 416:459 the mature cystine-knot growth factor domain after processing 416:476 We show that PDGFs share a conserved region in their prodomain sequences which can remain noncovalently associated with the mature cystine-knot growth factor domain after processing. 20534510 8 62 theme biochemical 1494:1504 arg1 data 1506:1509 The structural and biochemical data 1475:1509 The structural and biochemical data 1475:1509 The structural and biochemical data together offer insights into PDGF-PDGFR signaling, as well as strategies for PDGF-antagonism. 20534510 1 63 theme growth 205:210 arg1 factors 212:218 prototypic growth factors 194:218 prototypic growth factors 194:218 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 63 theme growth 205:210 arg1 factors 145:151 Platelet-derived growth factors 121:151 Platelet-derived growth factors (PDGFs) 121:159 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 1 63 theme growth 205:210 arg1 receptors 171:179 their receptors 165:179 their receptors (PDGFRs) 165:188 Platelet-derived growth factors (PDGFs) and their receptors (PDGFRs) are prototypic growth factors and receptor tyrosine kinases which have critical functions in development. 20534510 6 64 theme specific 1217:1224 arg1 conformation 1226:1237 a highly specific conformation 1208:1237 a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta 1208:1308 A large-scale structural organization and rearrangement is observed for PDGF-B upon receptor binding, in which the PDGF-B L1 loop, disordered in the structure of the free form, adopts a highly specific conformation to form hydrophobic interactions with the third Ig domain of PDGFRbeta. 20534510 3 65 theme association 562:572 arg1 mode 574:577 this conserved, hydrophobic association mode 534:577 this conserved, hydrophobic association mode 534:577 The structure of the PDGF-A/propeptide complex reveals this conserved, hydrophobic association mode. 20534510 4 66 theme first 648:652 arg1 PDGFRbeta 674:682 PDGFRbeta 674:682 PDGFRbeta 674:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 66 theme first 648:652 arg1 domains 663:669 the first three Ig domains 644:669 the first three Ig domains of PDGFRbeta 644:682 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 20534510 4 66 theme first 648:652 arg1 complex 617:623 the complex 613:623 the complex between PDGF-B 613:638 We also present the structure of the complex between PDGF-B and the first three Ig domains of PDGFRbeta, showing that two PDGF-B protomers clamp PDGFRbeta at their dimerization seam. 19276170 0 0 theme protein 77:83 arg1 protein 77:83 cartilage oligomeric matrix protein 49:83 cartilage oligomeric matrix protein 49:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 0 0 theme protein 77:83 arg1 domain 39:44 the signature domain 25:44 the signature domain of cartilage oligomeric matrix protein 25:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 1 1 theme Cartilage 153:161 arg1 important 261:269 important 261:269 important 261:269 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 1 theme Cartilage 153:161 arg1 thrombospondin-5 200:215 thrombospondin-5 200:215 thrombospondin-5 (TSP-5) 200:223 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 1 theme Cartilage 153:161 arg1 COMP 190:193 COMP 190:193 COMP 190:193 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 1 theme Cartilage 153:161 arg1 protein 181:187 Cartilage oligomeric matrix protein 153:187 Cartilage oligomeric matrix protein (COMP) 153:194 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 1 theme Cartilage 153:161 arg1 glycoprotein 240:251 a secreted glycoprotein 229:251 a secreted glycoprotein that is important for growth plate organization and function 229:312 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 6 2 theme potential 1210:1218 arg1 binding 1227:1233 potential ligand binding 1210:1233 potential ligand binding 1210:1233 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 0 3 theme matrix 70:75 arg1 protein 77:83 cartilage oligomeric matrix protein 49:83 cartilage oligomeric matrix protein 49:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 4 4 theme contiguous 776:785 arg1 series 787:792 a contiguous series 774:792 a contiguous series of calcium binding sites that associate with the CTD at multiple points 774:864 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 4 4 theme contiguous 776:785 arg1 repeats 762:768 the type 3 repeats 751:768 the type 3 repeats 751:768 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 1 5 theme oligomeric 163:172 arg1 important 261:269 important 261:269 important 261:269 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 5 theme oligomeric 163:172 arg1 thrombospondin-5 200:215 thrombospondin-5 200:215 thrombospondin-5 (TSP-5) 200:223 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 5 theme oligomeric 163:172 arg1 COMP 190:193 COMP 190:193 COMP 190:193 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 5 theme oligomeric 163:172 arg1 protein 181:187 Cartilage oligomeric matrix protein 153:187 Cartilage oligomeric matrix protein (COMP) 153:194 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 5 theme oligomeric 163:172 arg1 glycoprotein 240:251 a secreted glycoprotein 229:251 a secreted glycoprotein that is important for growth plate organization and function 229:312 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 3 6 theme 3.15-A 620:625 arg1 resolution 627:636 3.15-A resolution 620:636 3.15-A resolution 620:636 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 4 7 theme calcium 797:803 arg1 sites 813:817 calcium binding sites 797:817 calcium binding sites that associate with the CTD at multiple points 797:864 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 4 8 theme type 755:758 arg1 series 787:792 a contiguous series 774:792 a contiguous series of calcium binding sites that associate with the CTD at multiple points 774:864 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 4 8 theme type 755:758 arg1 repeats 762:768 the type 3 repeats 751:768 the type 3 repeats 751:768 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 5 9 theme crystal 871:877 arg1 packing 879:885 The crystal packing 867:885 The crystal packing 867:885 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 10 theme potential 906:914 arg1 site 1045:1048 a binding site 1035:1048 a binding site for collagens and other ligands 1035:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 10 theme potential 906:914 arg1 common 999:1004 common 999:1004 common 999:1004 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 10 theme potential 906:914 arg1 MIDAS 951:955 MIDAS 951:955 MIDAS 951:955 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 10 theme potential 906:914 arg1 site 945:948 an exposed potential metal-ion-dependent adhesion site 895:948 an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands 895:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 1 11 theme secreted 231:238 arg1 important 261:269 important 261:269 important 261:269 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 11 theme secreted 231:238 arg1 thrombospondin-5 200:215 thrombospondin-5 200:215 thrombospondin-5 (TSP-5) 200:223 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 11 theme secreted 231:238 arg1 protein 181:187 Cartilage oligomeric matrix protein 153:187 Cartilage oligomeric matrix protein (COMP) 153:194 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 11 theme secreted 231:238 arg1 glycoprotein 240:251 a secreted glycoprotein 229:251 a secreted glycoprotein that is important for growth plate organization and function 229:312 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 4 12 theme multiple 850:857 arg1 points 859:864 multiple points 850:864 multiple points 850:864 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 7 13 from structure 1240:1248 arg1 crystal 1305:1311 the crystal 1301:1311 the crystal 1301:1311 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 0 14 theme collagen 103:110 arg1 binding 144:150 collagen, glycosaminoglycan and integrin binding 103:150 collagen, glycosaminoglycan and integrin binding 103:150 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 2 15 from Mutations 315:323 arg1 COMP 328:331 COMP 328:331 COMP 328:331 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 3 16 theme recombinant 483:493 arg1 protein 495:501 a recombinant protein 481:501 a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography 481:667 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 1 17 theme matrix 174:179 arg1 important 261:269 important 261:269 important 261:269 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 17 theme matrix 174:179 arg1 thrombospondin-5 200:215 thrombospondin-5 200:215 thrombospondin-5 (TSP-5) 200:223 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 17 theme matrix 174:179 arg1 COMP 190:193 COMP 190:193 COMP 190:193 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 17 theme matrix 174:179 arg1 protein 181:187 Cartilage oligomeric matrix protein 153:187 Cartilage oligomeric matrix protein (COMP) 153:194 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 17 theme matrix 174:179 arg1 glycoprotein 240:251 a secreted glycoprotein 229:251 a secreted glycoprotein that is important for growth plate organization and function 229:312 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 6 18 theme bond 1152:1155 arg1 formation 1157:1165 disulfide bond formation 1142:1165 disulfide bond formation 1142:1165 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 3 19 theme protein 495:501 arg1 structure 468:476 the structure 464:476 the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography 464:667 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 0 20 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 0:151 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 3 21 theme X-ray 647:651 arg1 crystallography 653:667 X-ray crystallography 647:667 X-ray crystallography 647:667 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 6 22 theme disulfide 1142:1150 arg1 formation 1157:1165 disulfide bond formation 1142:1165 disulfide bond formation 1142:1165 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 3 23 theme epidermal 526:534 arg1 factor 543:548 the last epidermal growth factor 517:548 the last epidermal growth factor repeat 517:555 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 6 24 theme Disease-causing 1083:1097 arg1 mutations 1099:1107 Disease-causing mutations 1083:1107 Disease-causing mutations in COMP 1083:1115 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 1 25 gly glycoprotein 240:251 arg1 important 261:269 important 261:269 important 261:269 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 25 gly glycoprotein 240:251 arg1 thrombospondin-5 200:215 thrombospondin-5 200:215 thrombospondin-5 (TSP-5) 200:223 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 25 gly glycoprotein 240:251 arg1 protein 181:187 Cartilage oligomeric matrix protein 153:187 Cartilage oligomeric matrix protein (COMP) 153:194 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 1 25 gly glycoprotein 240:251 arg1 glycoprotein 240:251 a secreted glycoprotein 229:251 a secreted glycoprotein that is important for growth plate organization and function 229:312 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 3 26 theme growth 536:541 arg1 factor 543:548 the last epidermal growth factor 517:548 the last epidermal growth factor repeat 517:555 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 5 27 theme metal-ion-dependent 916:934 arg1 site 1045:1048 a binding site 1035:1048 a binding site for collagens and other ligands 1035:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 27 theme metal-ion-dependent 916:934 arg1 common 999:1004 common 999:1004 common 999:1004 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 27 theme metal-ion-dependent 916:934 arg1 MIDAS 951:955 MIDAS 951:955 MIDAS 951:955 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 27 theme metal-ion-dependent 916:934 arg1 site 945:948 an exposed potential metal-ion-dependent adhesion site 895:948 an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands 895:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 0 28 theme glycosaminoglycan 113:129 arg1 binding 144:150 collagen, glycosaminoglycan and integrin binding 103:150 collagen, glycosaminoglycan and integrin binding 103:150 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 7 29 theme unique 1273:1278 arg1 packing 1290:1296 its unique molecular packing 1269:1296 its unique molecular packing in the crystal 1269:1311 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 2 30 theme multiple 397:404 arg1 dysplasias 352:361 two skeletal dysplasias 339:361 two skeletal dysplasias 339:361 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 30 theme multiple 397:404 arg1 EDM1 428:431 EDM1 428:431 EDM1 428:431 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 30 theme multiple 397:404 arg1 dysplasia 417:425 multiple epiphyseal dysplasia 397:425 multiple epiphyseal dysplasia (EDM1) 397:432 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 31 theme epiphyseal 406:415 arg1 dysplasias 352:361 two skeletal dysplasias 339:361 two skeletal dysplasias 339:361 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 31 theme epiphyseal 406:415 arg1 EDM1 428:431 EDM1 428:431 EDM1 428:431 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 31 theme epiphyseal 406:415 arg1 dysplasia 417:425 multiple epiphyseal dysplasia 397:425 multiple epiphyseal dysplasia (EDM1) 397:432 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 5 32 theme other 1068:1072 arg1 ligands 1074:1080 other ligands 1068:1080 other ligands 1068:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 7 33 theme potential 1322:1330 arg1 sites 1344:1348 potential interactive sites 1322:1348 potential interactive sites 1322:1348 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 3 34 theme last 521:524 arg1 factor 543:548 the last epidermal growth factor 517:548 the last epidermal growth factor repeat 517:555 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 5 35 from site 945:948 arg1 edge 965:968 one edge 961:968 one edge of the beta-sandwich 961:989 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 0 36 theme signature 29:37 arg1 protein 77:83 cartilage oligomeric matrix protein 49:83 cartilage oligomeric matrix protein 49:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 0 36 theme signature 29:37 arg1 domain 39:44 the signature domain 25:44 the signature domain of cartilage oligomeric matrix protein 25:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 0 37 theme integrin 135:142 arg1 binding 144:150 collagen, glycosaminoglycan and integrin binding 103:150 collagen, glycosaminoglycan and integrin binding 103:150 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 5 38 theme beta-sandwich 977:989 arg1 edge 965:968 one edge 961:968 one edge of the beta-sandwich 961:989 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 7 39 theme molecular 1280:1288 arg1 packing 1290:1296 its unique molecular packing 1269:1296 its unique molecular packing in the crystal 1269:1311 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 7 40 from packing 1290:1296 arg1 crystal 1305:1311 the crystal 1301:1311 the crystal 1301:1311 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 1 41 theme growth 275:280 arg1 organization 288:299 growth plate organization 275:299 growth plate organization 275:299 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 6 42 theme ligand 1220:1225 arg1 binding 1227:1233 potential ligand binding 1210:1233 potential ligand binding 1210:1233 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 6 43 theme calcium 1125:1131 arg1 binding 1133:1139 calcium binding 1125:1139 calcium binding 1125:1139 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 1 44 theme plate 282:286 arg1 organization 288:299 growth plate organization 275:299 growth plate organization 275:299 Cartilage oligomeric matrix protein (COMP), or thrombospondin-5 (TSP-5), is a secreted glycoprotein that is important for growth plate organization and function. 19276170 4 45 theme sites 813:817 arg1 series 787:792 a contiguous series 774:792 a contiguous series of calcium binding sites that associate with the CTD at multiple points 774:864 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 4 45 theme sites 813:817 arg1 CTD 674:676 The CTD 670:676 The CTD 670:676 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 4 45 theme sites 813:817 arg1 beta-sandwich 683:695 a beta-sandwich 681:695 a beta-sandwich that is composed of 15 antiparallel beta-strands 681:744 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 4 45 theme sites 813:817 arg1 repeats 762:768 the type 3 repeats 751:768 the type 3 repeats 751:768 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 3 46 theme C-terminal 585:594 arg1 domain 596:601 the C-terminal domain 581:601 the C-terminal domain (CTD) of COMP to 3.15-A resolution 581:636 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 46 theme C-terminal 585:594 arg1 CTD 604:606 CTD 604:606 CTD 604:606 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 46 theme C-terminal 585:594 arg1 COMP 612:615 COMP 612:615 COMP to 3.15-A resolution 612:636 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 47 theme factor 543:548 arg1 repeat 550:555 the last epidermal growth factor repeat 517:555 the last epidermal growth factor repeat 517:555 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 0 48 theme domain 39:44 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 0:151 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 3 49 contain contains 508:515 arg1 protein 495:501 a recombinant protein 481:501 a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography 481:667 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 49 contain contains 508:515 arg2 repeat 550:555 the last epidermal growth factor repeat 517:555 the last epidermal growth factor repeat 517:555 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 4 50 theme binding 805:811 arg1 sites 813:817 calcium binding sites 797:817 calcium binding sites that associate with the CTD at multiple points 797:864 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 7 51 theme cartilage 1417:1425 arg1 structure 1427:1435 cartilage structure 1417:1435 cartilage structure 1417:1435 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 6 52 theme intramolecular 1168:1181 arg1 interactions 1183:1194 intramolecular interactions 1168:1194 intramolecular interactions 1168:1194 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 5 53 theme adhesion 936:943 arg1 site 1045:1048 a binding site 1035:1048 a binding site for collagens and other ligands 1035:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 53 theme adhesion 936:943 arg1 common 999:1004 common 999:1004 common 999:1004 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 53 theme adhesion 936:943 arg1 MIDAS 951:955 MIDAS 951:955 MIDAS 951:955 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 53 theme adhesion 936:943 arg1 site 945:948 an exposed potential metal-ion-dependent adhesion site 895:948 an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands 895:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 3 54 theme COMP 612:615 arg1 domain 596:601 the C-terminal domain 581:601 the C-terminal domain (CTD) of COMP to 3.15-A resolution 581:636 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 54 theme COMP 612:615 arg1 repeats 569:575 the type 3 repeats 558:575 the type 3 repeats 558:575 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 54 theme COMP 612:615 arg1 CTD 604:606 CTD 604:606 CTD 604:606 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 3 54 theme COMP 612:615 arg1 COMP 612:615 COMP 612:615 COMP to 3.15-A resolution 612:636 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 5 55 theme exposed 898:904 arg1 site 1045:1048 a binding site 1035:1048 a binding site for collagens and other ligands 1035:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 55 theme exposed 898:904 arg1 common 999:1004 common 999:1004 common 999:1004 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 55 theme exposed 898:904 arg1 MIDAS 951:955 MIDAS 951:955 MIDAS 951:955 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 55 theme exposed 898:904 arg1 site 945:948 an exposed potential metal-ion-dependent adhesion site 895:948 an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands 895:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 7 56 theme interactive 1332:1342 arg1 sites 1344:1348 potential interactive sites 1322:1348 potential interactive sites 1322:1348 The structure presented here and its unique molecular packing in the crystal identify potential interactive sites for glycosaminoglycans, integrins, and collagens, which are key to cartilage structure and function. 19276170 0 57 dep structure 12:20 arg1 implications 86:97 implications 86:97 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 0:151 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 3 58 dep contains 508:515 arg1 limit 638:642 limit 638:642 limit by X-ray crystallography 638:667 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 0 59 theme oligomeric 59:68 arg1 protein 77:83 cartilage oligomeric matrix protein 49:83 cartilage oligomeric matrix protein 49:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 6 60 from mutations 1099:1107 arg1 COMP 1112:1115 COMP 1112:1115 COMP 1112:1115 Disease-causing mutations in COMP disrupt calcium binding, disulfide bond formation, intramolecular interactions, or sites for potential ligand binding. 19276170 3 61 theme type 562:565 arg1 repeats 569:575 the type 3 repeats 558:575 the type 3 repeats 558:575 In this study, we determined the structure of a recombinant protein that contains the last epidermal growth factor repeat, the type 3 repeats and the C-terminal domain (CTD) of COMP to 3.15-A resolution limit by X-ray crystallography. 19276170 2 62 theme skeletal 343:350 arg1 pseudoachondroplasia 364:383 pseudoachondroplasia 364:383 pseudoachondroplasia (PSACH) 364:391 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 62 theme skeletal 343:350 arg1 dysplasias 352:361 two skeletal dysplasias 339:361 two skeletal dysplasias 339:361 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 2 62 theme skeletal 343:350 arg1 dysplasia 417:425 multiple epiphyseal dysplasia 397:425 multiple epiphyseal dysplasia (EDM1) 397:432 Mutations in COMP cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (EDM1). 19276170 0 63 theme cartilage 49:57 arg1 protein 77:83 cartilage oligomeric matrix protein 49:83 cartilage oligomeric matrix protein 49:83 The crystal structure of the signature domain of cartilage oligomeric matrix protein: implications for collagen, glycosaminoglycan and integrin binding. 19276170 4 64 theme antiparallel 720:731 arg1 beta-strands 733:744 15 antiparallel beta-strands 717:744 15 antiparallel beta-strands 717:744 The CTD is a beta-sandwich that is composed of 15 antiparallel beta-strands, and the type 3 repeats are a contiguous series of calcium binding sites that associate with the CTD at multiple points. 19276170 5 65 theme binding 1037:1043 arg1 site 1045:1048 a binding site 1035:1048 a binding site for collagens and other ligands 1035:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 19276170 5 65 theme binding 1037:1043 arg1 site 945:948 an exposed potential metal-ion-dependent adhesion site 895:948 an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands 895:1080 The crystal packing reveals an exposed potential metal-ion-dependent adhesion site (MIDAS) on one edge of the beta-sandwich that is common to all TSPs and may serve as a binding site for collagens and other ligands. 22718755 6 0 theme BMP9 856:859 arg1 interactions 871:882 BMP9 and BMP10 interactions 856:882 BMP9 and BMP10 interactions with ALK1 and type II receptors 856:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 10 1 theme low 1429:1431 arg1 receptors 1459:1467 low and high affinity type II receptors 1429:1467 low and high affinity type II receptors 1429:1467 In addition, the structure explains how BMP9 discriminates between low and high affinity type II receptors. 22718755 11 2 theme mechanistic 1522:1532 arg1 basis 1585:1589 a basis 1583:1589 a basis for novel anti-angiogenic therapies 1583:1625 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 11 2 theme mechanistic 1522:1532 arg1 insights 1534:1541 structural and mechanistic insights 1507:1541 structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies 1507:1625 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 5 3 theme TGF-β 642:646 arg1 BMP9 663:666 BMP9 663:666 BMP9 663:666 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 3 theme TGF-β 642:646 arg1 ligands 655:661 The TGF-β family ligands 638:661 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 3 theme TGF-β 642:646 arg1 BMP10 672:676 BMP10 672:676 BMP10 672:676 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 8 4 theme ternary 1090:1096 arg1 complex 1098:1104 a fully assembled ternary complex 1072:1104 a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB 1072:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 10 5 theme type 1451:1454 arg1 receptors 1459:1467 low and high affinity type II receptors 1429:1467 low and high affinity type II receptors 1429:1467 In addition, the structure explains how BMP9 discriminates between low and high affinity type II receptors. 22718755 6 6 with interactions 871:882 arg1 ALK1 889:892 ALK1 889:892 ALK1 889:892 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 6 6 with interactions 871:882 arg1 receptors 906:914 type II receptors 898:914 type II receptors 898:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 1 7 theme cell-specific 156:168 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 7 theme cell-specific 156:168 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 6 8 theme II 903:904 arg1 receptors 906:914 type II receptors 898:914 type II receptors 898:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 5 9 dep receptors 720:728 arg1 receptors 720:728 the three type II TGF-β family receptors 689:728 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 9 dep receptors 720:728 arg1 ActRIIB 739:745 ActRIIB 739:745 ActRIIB 739:745 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 9 dep receptors 720:728 arg1 BMPRII 752:757 BMPRII 752:757 BMPRII 752:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 9 dep receptors 720:728 arg1 ActRIIA 730:736 ActRIIA 730:736 ActRIIA 730:736 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 10 theme ALK1 783:786 arg1 signaling 788:796 ALK1 signaling 783:796 ALK1 signaling 783:796 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 8 11 theme extracellular 1123:1135 arg1 ActRIIB 1157:1163 ActRIIB 1157:1163 ActRIIB 1157:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 11 theme extracellular 1123:1135 arg1 ALK1 1148:1151 ALK1 1148:1151 ALK1 1148:1151 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 11 theme extracellular 1123:1135 arg1 domains 1137:1143 the extracellular domains 1119:1143 the extracellular domains of ALK1 and ActRIIB 1119:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 4 12 theme ALK1 622:625 arg1 signaling 627:635 ALK1 signaling 622:635 ALK1 signaling 622:635 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 7 13 theme receptor 990:997 arg1 binding 999:1005 type II receptor binding 982:1005 type II receptor binding 982:1005 Our data show that BMP9 displays a significant discrimination in type II receptor binding, whereas BMP10 does not. 22718755 7 14 theme type 982:985 arg1 receptor 990:997 type II receptor 982:997 type II receptor binding 982:1005 Our data show that BMP9 displays a significant discrimination in type II receptor binding, whereas BMP10 does not. 22718755 1 15 theme Activin 102:108 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 15 theme Activin 102:108 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 15 theme Activin 102:108 arg1 ALK1 134:137 ALK1 134:137 ALK1 134:137 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 15 theme Activin 102:108 arg1 regulator 228:236 an important regulator 215:236 an important regulator of normal blood vessel development as well as pathological tumor angiogenesis 215:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 6 16 theme BMP10 865:869 arg1 interactions 871:882 BMP9 and BMP10 interactions 856:882 BMP9 and BMP10 interactions with ALK1 and type II receptors 856:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 8 17 with complex 1098:1104 arg1 ActRIIB 1157:1163 ActRIIB 1157:1163 ActRIIB 1157:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 17 with complex 1098:1104 arg1 ALK1 1148:1151 ALK1 1148:1151 ALK1 1148:1151 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 17 with complex 1098:1104 arg1 domains 1137:1143 the extracellular domains 1119:1143 the extracellular domains of ALK1 and ActRIIB 1119:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 5 18 theme family 713:718 arg1 receptors 720:728 the three type II TGF-β family receptors 689:728 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 18 theme family 713:718 arg1 ActRIIB 739:745 ActRIIB 739:745 ActRIIB 739:745 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 18 theme family 713:718 arg1 BMPRII 752:757 BMPRII 752:757 BMPRII 752:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 18 theme family 713:718 arg1 ActRIIA 730:736 ActRIIA 730:736 ActRIIA 730:736 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 1 19 theme pathological 284:295 arg1 angiogenesis 303:314 pathological tumor angiogenesis 284:314 normal blood vessel development as well as pathological tumor angiogenesis 241:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 20 theme angiogenesis 303:314 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 20 theme angiogenesis 303:314 arg1 regulator 228:236 an important regulator 215:236 an important regulator of normal blood vessel development as well as pathological tumor angiogenesis 215:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 0 21 theme ALK1 77:80 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 9 22 theme ALK1 1217:1220 arg1 specificity 1202:1212 the high specificity 1193:1212 the high specificity of ALK1 for BMP9/10 1193:1232 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 2 23 theme therapeutic 347:357 arg1 target 359:364 an important therapeutic target 334:364 an important therapeutic target 334:364 As such, ALK1 is an important therapeutic target. 22718755 2 23 theme therapeutic 347:357 arg1 ALK1 326:329 ALK1 326:329 ALK1 326:329 As such, ALK1 is an important therapeutic target. 22718755 0 24 theme signaling 83:91 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 9 25 theme novel 1253:1257 arg1 orientation 1259:1269 a novel orientation 1251:1269 a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions 1251:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 0 26 theme kinase 67:72 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 3 27 theme several 373:379 arg1 agents 395:400 several ALK1-directed agents 373:400 several ALK1-directed agents 373:400 Thus, several ALK1-directed agents are currently in clinical trials as anti-angiogenic cancer therapeutics. 22718755 3 28 theme clinical 419:426 arg1 trials 428:433 clinical trials 419:433 clinical trials 419:433 Thus, several ALK1-directed agents are currently in clinical trials as anti-angiogenic cancer therapeutics. 22718755 1 29 theme blood 248:252 arg1 development 261:271 normal blood vessel development 241:271 normal blood vessel development as well as pathological tumor angiogenesis 241:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 4 30 theme signaling 532:540 arg1 pathway 542:548 the ALK1 signaling pathway 523:548 the ALK1 signaling pathway 523:548 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 1 31 theme development 261:271 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 31 theme development 261:271 arg1 regulator 228:236 an important regulator 215:236 an important regulator of normal blood vessel development as well as pathological tumor angiogenesis 215:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 0 32 theme receptor-like 53:65 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 3 33 theme cancer 454:459 arg1 therapeutics 461:472 anti-angiogenic cancer therapeutics 438:472 anti-angiogenic cancer therapeutics 438:472 Thus, several ALK1-directed agents are currently in clinical trials as anti-angiogenic cancer therapeutics. 22718755 0 34 theme affinity 36:43 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 9 35 theme ALK1 1274:1277 arg1 orientation 1259:1269 a novel orientation 1251:1269 a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions 1251:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 7 36 from discrimination 964:977 arg1 binding 999:1005 type II receptor binding 982:1005 type II receptor binding 982:1005 Our data show that BMP9 displays a significant discrimination in type II receptor binding, whereas BMP10 does not. 22718755 10 37 theme affinity 1442:1449 arg1 receptors 1459:1467 low and high affinity type II receptors 1429:1467 low and high affinity type II receptors 1429:1467 In addition, the structure explains how BMP9 discriminates between low and high affinity type II receptors. 22718755 11 38 theme anti-angiogenic 1601:1615 arg1 therapies 1617:1625 novel anti-angiogenic therapies 1595:1625 novel anti-angiogenic therapies 1595:1625 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 1 39 theme superfamily 199:209 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 39 theme superfamily 199:209 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 4 40 theme biophysical 578:588 arg1 basis 605:609 the biophysical and structural basis 574:609 the biophysical and structural basis underlying ALK1 signaling 574:635 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 11 41 theme ALK1 1548:1551 arg1 signaling 1553:1561 ALK1 signaling 1548:1561 ALK1 signaling 1548:1561 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 9 42 theme high 1197:1200 arg1 specificity 1202:1212 the high specificity 1193:1212 the high specificity of ALK1 for BMP9/10 1193:1232 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 1 43 theme important 218:226 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 43 theme important 218:226 arg1 regulator 228:236 an important regulator 215:236 an important regulator of normal blood vessel development as well as pathological tumor angiogenesis 215:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 6 44 theme thermodynamic 830:842 arg1 analysis 844:851 a kinetic and thermodynamic analysis 816:851 a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors 816:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 10 45 theme II 1456:1457 arg1 receptors 1459:1467 low and high affinity type II receptors 1429:1467 low and high affinity type II receptors 1429:1467 In addition, the structure explains how BMP9 discriminates between low and high affinity type II receptors. 22718755 9 46 theme unique 1318:1323 arg1 set 1325:1327 a unique set 1316:1327 a unique set of receptor-ligand interactions 1316:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 9 46 theme unique 1318:1323 arg1 interactions 1348:1359 receptor-ligand interactions 1332:1359 receptor-ligand interactions 1332:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 8 47 theme assembled 1080:1088 arg1 complex 1098:1104 a fully assembled ternary complex 1072:1104 a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB 1072:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 6 48 theme kinetic 818:824 arg1 analysis 844:851 a kinetic and thermodynamic analysis 816:851 a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors 816:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 8 49 theme ALK1 1148:1151 arg1 ActRIIB 1157:1163 ActRIIB 1157:1163 ActRIIB 1157:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 49 theme ALK1 1148:1151 arg1 ALK1 1148:1151 ALK1 1148:1151 ALK1 1148:1151 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 49 theme ALK1 1148:1151 arg1 domains 1137:1143 the extracellular domains 1119:1143 the extracellular domains of ALK1 and ActRIIB 1119:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 1 50 theme endothelial 144:154 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 50 theme endothelial 144:154 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 8 51 theme complex 1098:1104 arg1 structure 1059:1067 the crystal structure 1047:1067 the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB 1047:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 52 theme ActRIIB 1157:1163 arg1 ActRIIB 1157:1163 ActRIIB 1157:1163 ActRIIB 1157:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 52 theme ActRIIB 1157:1163 arg1 ALK1 1148:1151 ALK1 1148:1151 ALK1 1148:1151 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 8 52 theme ActRIIB 1157:1163 arg1 domains 1137:1143 the extracellular domains 1119:1143 the extracellular domains of ALK1 and ActRIIB 1119:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 9 53 theme interactions 1348:1359 arg1 set 1325:1327 a unique set 1316:1327 a unique set of receptor-ligand interactions 1316:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 9 53 theme interactions 1348:1359 arg1 interactions 1348:1359 receptor-ligand interactions 1332:1359 receptor-ligand interactions 1332:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 8 54 theme BMP9 1109:1112 arg1 complex 1098:1104 a fully assembled ternary complex 1072:1104 a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB 1072:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 6 55 theme type 898:901 arg1 receptors 906:914 type II receptors 898:914 type II receptors 898:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 4 56 theme structural 594:603 arg1 basis 605:609 the biophysical and structural basis 574:609 the biophysical and structural basis underlying ALK1 signaling 574:635 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 9 57 theme receptor-ligand 1332:1346 arg1 interactions 1348:1359 receptor-ligand interactions 1332:1359 receptor-ligand interactions 1332:1359 The structure reveals that the high specificity of ALK1 for BMP9/10 is determined by a novel orientation of ALK1 with respect to BMP9, which leads to a unique set of receptor-ligand interactions. 22718755 5 58 theme family 648:653 arg1 BMP9 663:666 BMP9 663:666 BMP9 663:666 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 58 theme family 648:653 arg1 ligands 655:661 The TGF-β family ligands 638:661 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 58 theme family 648:653 arg1 BMP10 672:676 BMP10 672:676 BMP10 672:676 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 6 59 theme interactions 871:882 arg1 analysis 844:851 a kinetic and thermodynamic analysis 816:851 a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors 816:914 Here, we provide a kinetic and thermodynamic analysis of BMP9 and BMP10 interactions with ALK1 and type II receptors. 22718755 7 60 theme significant 952:962 arg1 discrimination 964:977 a significant discrimination 950:977 a significant discrimination in type II receptor binding 950:1005 Our data show that BMP9 displays a significant discrimination in type II receptor binding, whereas BMP10 does not. 22718755 1 61 theme receptor-like 110:122 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 61 theme receptor-like 110:122 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 61 theme receptor-like 110:122 arg1 ALK1 134:137 ALK1 134:137 ALK1 134:137 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 61 theme receptor-like 110:122 arg1 regulator 228:236 an important regulator 215:236 an important regulator of normal blood vessel development as well as pathological tumor angiogenesis 215:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 5 62 theme TGF-β 707:711 arg1 receptors 720:728 the three type II TGF-β family receptors 689:728 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 62 theme TGF-β 707:711 arg1 ActRIIB 739:745 ActRIIB 739:745 ActRIIB 739:745 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 62 theme TGF-β 707:711 arg1 BMPRII 752:757 BMPRII 752:757 BMPRII 752:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 62 theme TGF-β 707:711 arg1 ActRIIA 730:736 ActRIIA 730:736 ActRIIA 730:736 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 1 63 theme type 170:173 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 63 theme type 170:173 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 64 theme tumor 297:301 arg1 angiogenesis 303:314 pathological tumor angiogenesis 284:314 normal blood vessel development as well as pathological tumor angiogenesis 241:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 4 65 theme biological 485:494 arg1 importance 509:518 the biological and clinical importance 481:518 the biological and clinical importance of the ALK1 signaling pathway 481:548 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 0 66 theme complex 93:99 arg1 structure 16:24 structure 16:24 structure 16:24 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 0 66 theme complex 93:99 arg1 Specificity 0:10 Specificity 0:10 Specificity 0:10 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 1 67 theme I 175:175 arg1 kinase 124:129 Activin receptor-like kinase 1 102:131 Activin receptor-like kinase 1 (ALK1) 102:138 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 1 67 theme I 175:175 arg1 receptor 177:184 an endothelial cell-specific type I receptor 141:184 an endothelial cell-specific type I receptor of the TGF-β superfamily 141:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 5 68 dep ligands 655:661 arg1 BMP9 663:666 BMP9 663:666 BMP9 663:666 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 68 dep ligands 655:661 arg1 ligands 655:661 The TGF-β family ligands 638:661 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 68 dep ligands 655:661 arg1 BMP10 672:676 BMP10 672:676 BMP10 672:676 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 7 69 dep displays 941:948 arg1 whereas 1008:1014 whereas 1008:1014 whereas 1008:1014 Our data show that BMP9 displays a significant discrimination in type II receptor binding, whereas BMP10 does not. 22718755 2 70 theme important 337:345 arg1 target 359:364 an important therapeutic target 334:364 an important therapeutic target 334:364 As such, ALK1 is an important therapeutic target. 22718755 2 70 theme important 337:345 arg1 ALK1 326:329 ALK1 326:329 ALK1 326:329 As such, ALK1 is an important therapeutic target. 22718755 4 71 theme ALK1 527:530 arg1 pathway 542:548 the ALK1 signaling pathway 523:548 the ALK1 signaling pathway 523:548 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 3 72 theme ALK1-directed 381:393 arg1 agents 395:400 several ALK1-directed agents 373:400 several ALK1-directed agents 373:400 Thus, several ALK1-directed agents are currently in clinical trials as anti-angiogenic cancer therapeutics. 22718755 1 73 theme normal 241:246 arg1 development 261:271 normal blood vessel development 241:271 normal blood vessel development as well as pathological tumor angiogenesis 241:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 4 74 theme pathway 542:548 arg1 importance 509:518 the biological and clinical importance 481:518 the biological and clinical importance of the ALK1 signaling pathway 481:548 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 0 75 theme high 31:34 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 1 76 theme vessel 254:259 arg1 development 261:271 normal blood vessel development 241:271 normal blood vessel development as well as pathological tumor angiogenesis 241:314 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 4 77 theme clinical 500:507 arg1 importance 509:518 the biological and clinical importance 481:518 the biological and clinical importance of the ALK1 signaling pathway 481:548 Given the biological and clinical importance of the ALK1 signaling pathway, we sought to elucidate the biophysical and structural basis underlying ALK1 signaling. 22718755 0 78 theme activin 45:51 arg1 complex 93:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 a high affinity activin receptor-like kinase 1 (ALK1) signaling complex 29:99 Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. 22718755 5 79 theme type 699:702 arg1 receptors 720:728 the three type II TGF-β family receptors 689:728 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII 638:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 79 theme type 699:702 arg1 ActRIIB 739:745 ActRIIB 739:745 ActRIIB 739:745 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 79 theme type 699:702 arg1 BMPRII 752:757 BMPRII 752:757 BMPRII 752:757 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 5 79 theme type 699:702 arg1 ActRIIA 730:736 ActRIIA 730:736 ActRIIA 730:736 The TGF-β family ligands BMP9 and BMP10 as well as the three type II TGF-β family receptors ActRIIA, ActRIIB, and BMPRII have been implicated in ALK1 signaling. 22718755 11 80 theme novel 1595:1599 arg1 therapies 1617:1625 novel anti-angiogenic therapies 1595:1625 novel anti-angiogenic therapies 1595:1625 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 11 81 theme structural 1507:1516 arg1 basis 1585:1589 a basis 1583:1589 a basis for novel anti-angiogenic therapies 1583:1625 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 11 81 theme structural 1507:1516 arg1 insights 1534:1541 structural and mechanistic insights 1507:1541 structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies 1507:1625 Taken together, our findings provide structural and mechanistic insights into ALK1 signaling that could serve as a basis for novel anti-angiogenic therapies. 22718755 3 82 theme anti-angiogenic 438:452 arg1 therapeutics 461:472 anti-angiogenic cancer therapeutics 438:472 anti-angiogenic cancer therapeutics 438:472 Thus, several ALK1-directed agents are currently in clinical trials as anti-angiogenic cancer therapeutics. 22718755 1 83 theme TGF-β 193:197 arg1 superfamily 199:209 the TGF-β superfamily 189:209 the TGF-β superfamily 189:209 Activin receptor-like kinase 1 (ALK1), an endothelial cell-specific type I receptor of the TGF-β superfamily, is an important regulator of normal blood vessel development as well as pathological tumor angiogenesis. 22718755 8 84 theme crystal 1051:1057 arg1 structure 1059:1067 the crystal structure 1047:1067 the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB 1047:1163 We also report the crystal structure of a fully assembled ternary complex of BMP9 with the extracellular domains of ALK1 and ActRIIB. 22718755 10 85 theme high 1437:1440 arg1 receptors 1459:1467 low and high affinity type II receptors 1429:1467 low and high affinity type II receptors 1429:1467 In addition, the structure explains how BMP9 discriminates between low and high affinity type II receptors. 18488039 0 0 theme receptor 76:83 arg1 ephrin-B2 85:93 their cell-surface receptor ephrin-B2 57:93 their cell-surface receptor ephrin-B2 57:93 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 2 1 theme cell-surface 360:371 arg1 receptors 373:381 the cell-surface receptors 356:381 the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3) 356:421 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 1 theme cell-surface 360:371 arg1 ephrin-B3 405:413 ephrin-B3 405:413 ephrin-B3 (EFNB3) 405:421 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 1 theme cell-surface 360:371 arg1 ephrin-B2 383:391 ephrin-B2 383:391 ephrin-B2 (EFNB2) 383:399 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 4 2 theme attachment 584:593 arg1 complexes 595:603 previously solved paramyxovirus attachment complexes 552:603 previously solved paramyxovirus attachment complexes 552:603 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 0 3 theme cell-surface 63:74 arg1 ephrin-B2 85:93 their cell-surface receptor ephrin-B2 57:93 their cell-surface receptor ephrin-B2 57:93 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 3 4 from glycoproteins 494:506 arg1 complex 511:517 complex 511:517 complex with human EFNB2 511:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 5 5 theme antivirals 897:906 arg1 development 882:892 the development 878:892 the development of antivirals against sialic acid binding viruses 878:942 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 4 6 theme paramyxovirus 570:582 arg1 complexes 595:603 previously solved paramyxovirus attachment complexes 552:603 previously solved paramyxovirus attachment complexes 552:603 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 3 7 with complex 511:517 arg1 EFNB2 530:534 human EFNB2 524:534 human EFNB2 524:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 4 8 from chain 782:786 arg1 EFNB2 791:795 EFNB2 791:795 EFNB2 that fits snugly into a hydrophobic pocket on the viral protein 791:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 4 9 theme side 777:780 arg1 chain 782:786 a crucial phenylalanine side chain 753:786 a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein 753:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 1 10 theme Biosafety 259:267 arg1 pathogens 277:285 Biosafety Level 4 pathogens 259:285 Biosafety Level 4 pathogens 259:285 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 4 11 theme phenylalanine 763:775 arg1 chain 782:786 a crucial phenylalanine side chain 753:786 a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein 753:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 3 12 from complex 511:517 arg1 structures 447:456 crystal structures 439:456 crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 439:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 3 13 theme glycoproteins 494:506 arg1 structures 447:456 crystal structures 439:456 crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 439:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 1 14 theme Level 269:273 arg1 pathogens 277:285 Biosafety Level 4 pathogens 259:285 Biosafety Level 4 pathogens 259:285 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 3 15 theme Hendra 476:481 arg1 glycoproteins 494:506 both Nipah and Hendra attachment glycoproteins 461:506 both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 461:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 4 16 theme sialic 628:633 arg1 interactions 640:651 sialic acid interactions 628:651 sialic acid interactions 628:651 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 3 17 theme human 524:528 arg1 EFNB2 530:534 human EFNB2 524:534 human EFNB2 524:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 4 18 theme Hendra 668:673 arg1 complexes 675:683 the Nipah and Hendra complexes 654:683 the Nipah and Hendra complexes 654:683 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 4 19 theme Nipah 658:662 arg1 complexes 675:683 the Nipah and Hendra complexes 654:683 the Nipah and Hendra complexes 654:683 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 2 20 dep receptors 373:381 arg1 receptors 373:381 the cell-surface receptors 356:381 the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3) 356:421 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 20 dep receptors 373:381 arg1 EFNB3 416:420 EFNB3 416:420 EFNB3 416:420 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 20 dep receptors 373:381 arg1 EFNB2 394:398 EFNB2 394:398 EFNB2 394:398 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 20 dep receptors 373:381 arg1 ephrin-B3 405:413 ephrin-B3 405:413 ephrin-B3 (EFNB3) 405:421 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 20 dep receptors 373:381 arg1 ephrin-B2 383:391 ephrin-B2 383:391 ephrin-B2 (EFNB2) 383:399 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 2 21 gly glycoproteins 305:317 arg1 glycoproteins 305:317 Their attachment glycoproteins 288:317 Their attachment glycoproteins 288:317 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 1 22 theme rapid 184:188 arg1 onset 190:194 rapid onset 184:194 rapid onset 184:194 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 0 23 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 0:94 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 4 24 theme protein-protein 716:730 arg1 interface 732:740 an extensive protein-protein interface 703:740 an extensive protein-protein interface 703:740 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 0 25 theme Nipah 20:24 arg1 attachment 43:52 Nipah and Hendra virus attachment 20:52 Nipah and Hendra virus attachment 20:52 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 5 26 theme binding 928:934 arg1 viruses 936:942 sialic acid binding viruses 916:942 sialic acid binding viruses 916:942 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 1 27 theme Nipah 96:100 arg1 viruses 113:119 Nipah and Hendra viruses 96:119 Nipah and Hendra viruses 96:119 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 1 27 theme Nipah 96:100 arg1 paramyxoviruses 134:148 emergent paramyxoviruses 125:148 emergent paramyxoviruses 125:148 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 4 28 theme acid 635:638 arg1 interactions 640:651 sialic acid interactions 628:651 sialic acid interactions 628:651 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 4 29 theme solved 563:568 arg1 complexes 595:603 previously solved paramyxovirus attachment complexes 552:603 previously solved paramyxovirus attachment complexes 552:603 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 5 30 theme structural 969:978 arg1 template 980:987 a structural template 967:987 a structural template to target antiviral inhibition of protein-protein interactions 967:1050 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 2 31 theme attachment 294:303 arg1 glycoproteins 305:317 Their attachment glycoproteins 288:317 Their attachment glycoproteins 288:317 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 18488039 1 32 theme high 200:203 arg1 rates 215:219 high mortality rates 200:219 high mortality rates 200:219 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 0 33 theme Hendra 30:35 arg1 attachment 43:52 Nipah and Hendra virus attachment 20:52 Nipah and Hendra virus attachment 20:52 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 5 34 theme sialic 916:921 arg1 viruses 936:942 sialic acid binding viruses 916:942 sialic acid binding viruses 916:942 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 3 35 theme crystal 439:445 arg1 structures 447:456 crystal structures 439:456 crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 439:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 1 36 theme mortality 205:213 arg1 rates 215:219 high mortality rates 200:219 high mortality rates 200:219 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 5 37 theme protein-protein 1023:1037 arg1 interactions 1039:1050 protein-protein interactions 1023:1050 protein-protein interactions 1023:1050 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 4 38 from pocket 833:838 arg1 protein 853:859 the viral protein 843:859 the viral protein 843:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 0 39 theme attachment 43:52 arg1 basis 11:15 Structural basis 0:15 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 0:94 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 5 40 with analogy 865:871 arg1 development 882:892 the development 878:892 the development of antivirals against sialic acid binding viruses 878:942 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 5 41 theme interactions 1039:1050 arg1 inhibition 1009:1018 antiviral inhibition 999:1018 antiviral inhibition of protein-protein interactions 999:1050 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 3 42 from structures 447:456 arg1 complex 511:517 complex 511:517 complex with human EFNB2 511:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 1 43 theme Hendra 106:111 arg1 viruses 113:119 Nipah and Hendra viruses 96:119 Nipah and Hendra viruses 96:119 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 1 43 theme Hendra 106:111 arg1 paramyxoviruses 134:148 emergent paramyxoviruses 125:148 emergent paramyxoviruses 125:148 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 0 44 theme virus 37:41 arg1 attachment 43:52 Nipah and Hendra virus attachment 20:52 Nipah and Hendra virus attachment 20:52 Structural basis of Nipah and Hendra virus attachment to their cell-surface receptor ephrin-B2. 18488039 4 45 theme hydrophobic 821:831 arg1 pocket 833:838 a hydrophobic pocket 819:838 a hydrophobic pocket on the viral protein 819:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 3 46 theme attachment 483:492 arg1 glycoproteins 494:506 both Nipah and Hendra attachment glycoproteins 461:506 both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 461:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 4 47 theme extensive 706:714 arg1 interface 732:740 an extensive protein-protein interface 703:740 an extensive protein-protein interface 703:740 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 3 48 theme Nipah 466:470 arg1 glycoproteins 494:506 both Nipah and Hendra attachment glycoproteins 461:506 both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 461:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 4 49 theme crucial 755:761 arg1 chain 782:786 a crucial phenylalanine side chain 753:786 a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein 753:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 3 50 gly glycoproteins 494:506 arg1 glycoproteins 494:506 both Nipah and Hendra attachment glycoproteins 461:506 both Nipah and Hendra attachment glycoproteins in complex with human EFNB2 461:534 Here we report crystal structures of both Nipah and Hendra attachment glycoproteins in complex with human EFNB2. 18488039 1 51 theme emergent 125:132 arg1 viruses 113:119 Nipah and Hendra viruses 96:119 Nipah and Hendra viruses 96:119 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 1 51 theme emergent 125:132 arg1 paramyxoviruses 134:148 emergent paramyxoviruses 125:148 emergent paramyxoviruses 125:148 Nipah and Hendra viruses are emergent paramyxoviruses, causing disease characterized by rapid onset and high mortality rates, resulting in their classification as Biosafety Level 4 pathogens. 18488039 5 52 theme acid 923:926 arg1 viruses 936:942 sialic acid binding viruses 916:942 sialic acid binding viruses 916:942 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 5 53 theme antiviral 999:1007 arg1 inhibition 1009:1018 antiviral inhibition 999:1018 antiviral inhibition of protein-protein interactions 999:1050 By analogy with the development of antivirals against sialic acid binding viruses, these results provide a structural template to target antiviral inhibition of protein-protein interactions. 18488039 4 54 theme viral 847:851 arg1 protein 853:859 the viral protein 843:859 the viral protein 843:859 In contrast to previously solved paramyxovirus attachment complexes, which are mediated by sialic acid interactions, the Nipah and Hendra complexes are maintained by an extensive protein-protein interface, including a crucial phenylalanine side chain on EFNB2 that fits snugly into a hydrophobic pocket on the viral protein. 18488039 2 55 theme receptors 373:381 arg1 recognition 341:351 the recognition 337:351 the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3) 337:421 Their attachment glycoproteins are essential for the recognition of the cell-surface receptors ephrin-B2 (EFNB2) and ephrin-B3 (EFNB3). 19836338 4 0 theme class 608:612 arg1 ligands 616:622 class B ligands 608:622 class B ligands 608:622 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 1 1 theme physiological 153:165 arg1 processes 167:175 physiological processes 153:175 physiological processes 153:175 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 5 2 theme reported 668:675 arg1 structures 690:699 previously reported Eph receptor structures 657:699 previously reported Eph receptor structures 657:699 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 3 theme structural 830:839 arg1 hallmarks 841:849 structural hallmarks 830:849 structural hallmarks of the class B Eph receptors 830:878 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 4 theme binding 805:811 arg1 ephrinB2 813:820 binding ephrinB2 805:820 binding ephrinB2 805:820 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 1 5 theme processes 167:175 arg1 array 144:148 an array 141:148 an array of physiological processes 141:175 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 1 6 theme A 246:246 arg1 ephrins 260:266 both A and B class ephrins 241:266 both A and B class ephrins 241:266 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 6 7 theme receptor 987:994 arg1 conformations 996:1008 both A and B class Eph receptor conformations 964:1008 both A and B class Eph receptor conformations 964:1008 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 2 8 dep ephrinA2 407:414 arg1 2 404:404 2 404:404 2 404:404 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 6 9 theme cross-class 1061:1071 arg1 reactivity 1073:1082 EphA-type cross-class reactivity 1051:1082 EphA-type cross-class reactivity 1051:1082 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 3 10 theme conformational 473:486 arg1 plasticity 488:497 significant conformational plasticity 461:497 significant conformational plasticity 461:497 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 5 11 theme Eph 866:868 arg1 receptors 870:878 the class B Eph receptors 854:878 the class B Eph receptors 854:878 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 3 12 theme ligand 506:511 arg1 face 521:524 its ligand binding face 502:524 its ligand binding face 502:524 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 2 13 with complex 373:379 arg1 ephrinB2 390:397 (1) ephrinB2 386:397 (1) ephrinB2 386:397 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 2 13 with complex 373:379 arg1 ephrinA2 407:414 (2) ephrinA2 403:414 (2) ephrinA2 403:414 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 3 14 theme structures 429:438 arg1 set 422:424 This set 417:424 This set of structures 417:438 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 3 14 theme structures 429:438 arg1 structures 429:438 structures 429:438 structures 429:438 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 1 15 theme B 252:252 arg1 ephrins 260:266 both A and B class ephrins 241:266 both A and B class ephrins 241:266 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 4 16 theme B 614:614 arg1 ligands 616:622 class B ligands 608:622 class B ligands 608:622 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 5 17 theme class 858:862 arg1 receptors 870:878 the class B Eph receptors 854:878 the class B Eph receptors 854:878 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 6 18 theme class 977:981 arg1 conformations 996:1008 both A and B class Eph receptor conformations 964:1008 both A and B class Eph receptor conformations 964:1008 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 5 19 theme Eph 784:786 arg1 receptors 788:796 other class A Eph receptors 770:796 other class A Eph receptors 770:796 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 20 theme class 776:780 arg1 receptors 788:796 other class A Eph receptors 770:796 other class A Eph receptors 770:796 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 4 21 theme In 527:528 arg1 data 544:547 In vitro binding data 527:547 In vitro binding data 527:547 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 3 22 theme significant 461:471 arg1 plasticity 488:497 significant conformational plasticity 461:497 significant conformational plasticity 461:497 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 5 23 with complex 738:744 arg1 ephrinA2 751:758 ephrinA2 751:758 ephrinA2 751:758 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 24 theme A 782:782 arg1 receptors 788:796 other class A Eph receptors 770:796 other class A Eph receptors 770:796 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 6 25 theme EphA-type 1051:1059 arg1 reactivity 1073:1082 EphA-type cross-class reactivity 1051:1082 EphA-type cross-class reactivity 1051:1082 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 6 26 theme interactive 886:896 arg1 plasticity 898:907 This interactive plasticity 881:907 This interactive plasticity 881:907 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 1 27 theme EphA4 87:91 arg1 kinase 102:107 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 6 28 theme Eph 983:985 arg1 conformations 996:1008 both A and B class Eph receptor conformations 964:1008 both A and B class Eph receptor conformations 964:1008 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 2 29 theme domain 353:358 arg1 structure 315:323 the crystal structure 303:323 the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2 303:414 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 0 30 theme Structural 0:9 arg1 plasticity 11:20 Structural plasticity 0:20 Structural plasticity of eph receptor A4 0:39 Structural plasticity of eph receptor A4 facilitates cross-class ephrin signaling. 19836338 1 31 theme class 254:258 arg1 ephrins 260:266 both A and B class ephrins 241:266 both A and B class ephrins 241:266 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 2 32 dep ephrinB2 390:397 arg1 1 387:387 1 387:387 1 387:387 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 1 33 theme tyrosine 93:100 arg1 kinase 102:107 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 2 34 theme binding 345:351 arg1 domain 353:358 the EphA4 ligand binding domain 328:358 the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2 328:414 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 0 35 theme eph 25:27 arg1 A4 38:39 eph receptor A4 25:39 eph receptor A4 25:39 Structural plasticity of eph receptor A4 facilitates cross-class ephrin signaling. 19836338 6 36 theme molecular 1031:1039 arg1 basis 1041:1045 a molecular basis 1029:1045 a molecular basis for EphA-type cross-class reactivity 1029:1082 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 5 37 theme B 864:864 arg1 receptors 870:878 the class B Eph receptors 854:878 the class B Eph receptors 854:878 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 3 38 theme binding 513:519 arg1 face 521:524 its ligand binding face 502:524 its ligand binding face 502:524 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 2 39 theme ligand 338:343 arg1 domain 353:358 the EphA4 ligand binding domain 328:358 the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2 328:414 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 1 40 theme cell 109:112 arg1 receptor 122:129 cell surface receptor 109:129 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 1 41 theme known 203:207 arg1 receptor 221:228 the only currently known class A Eph receptor 184:228 the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity 184:285 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 0 42 theme A4 38:39 arg1 plasticity 11:20 Structural plasticity 0:20 Structural plasticity of eph receptor A4 0:39 Structural plasticity of eph receptor A4 facilitates cross-class ephrin signaling. 19836338 1 43 theme high 273:276 arg1 affinity 278:285 high affinity 273:285 high affinity 273:285 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 4 44 theme higher 575:580 arg1 affinity 582:589 a higher affinity 573:589 a higher affinity for class A than class B ligands 573:622 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 4 45 theme class 595:599 arg1 A 601:601 class A 595:601 class A 595:601 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 1 46 theme class 209:213 arg1 receptor 221:228 the only currently known class A Eph receptor 184:228 the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity 184:285 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 0 47 theme receptor 29:36 arg1 A4 38:39 eph receptor A4 25:39 eph receptor A4 25:39 Structural plasticity of eph receptor A4 facilitates cross-class ephrin signaling. 19836338 1 48 theme A 215:215 arg1 receptor 221:228 the only currently known class A Eph receptor 184:228 the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity 184:285 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 0 49 theme cross-class 53:63 arg1 signaling 72:80 cross-class ephrin signaling 53:80 cross-class ephrin signaling 53:80 Structural plasticity of eph receptor A4 facilitates cross-class ephrin signaling. 19836338 3 50 contain has 457:459 arg2 plasticity 488:497 significant conformational plasticity 461:497 significant conformational plasticity 461:497 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 3 50 contain has 457:459 arg1 EphA4 451:455 EphA4 451:455 EphA4 451:455 This set of structures shows that EphA4 has significant conformational plasticity in its ligand binding face. 19836338 6 51 theme able 950:953 arg1 EphA4 917:921 EphA4 917:921 EphA4 917:921 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 6 51 theme able 950:953 arg1 chameleon 939:947 a structural chameleon 926:947 a structural chameleon 926:947 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 4 52 contain has 569:571 arg1 it 566:567 it 566:567 it 566:567 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 4 52 contain has 569:571 arg2 affinity 582:589 a higher affinity 573:589 a higher affinity for class A than class B ligands 573:622 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 1 53 theme surface 114:120 arg1 receptor 122:129 cell surface receptor 109:129 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 6 54 theme A 969:969 arg1 conformations 996:1008 both A and B class Eph receptor conformations 964:1008 both A and B class Eph receptor conformations 964:1008 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 2 55 theme crystal 307:313 arg1 structure 315:323 the crystal structure 303:323 the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2 303:414 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 19836338 5 56 dep resembles 760:768 arg1 assumes 822:828 assumes 822:828 assumes structural hallmarks of the class B Eph receptors 822:878 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 57 theme receptor 681:688 arg1 structures 690:699 previously reported Eph receptor structures 657:699 previously reported Eph receptor structures 657:699 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 1 58 theme Eph 217:219 arg1 receptor 221:228 the only currently known class A Eph receptor 184:228 the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity 184:285 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 4 59 dep In 527:528 arg1 vitro 530:534 vitro 530:534 vitro 530:534 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 5 60 theme receptors 870:878 arg1 hallmarks 841:849 structural hallmarks 830:849 structural hallmarks of the class B Eph receptors 830:878 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 61 theme Structural 625:634 arg1 analyses 636:643 Structural analyses 625:643 Structural analyses 625:643 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 4 62 theme binding 536:542 arg1 data 544:547 In vitro binding data 527:547 In vitro binding data 527:547 In vitro binding data demonstrate that it has a higher affinity for class A than class B ligands. 19836338 5 63 theme Eph 677:679 arg1 structures 690:699 previously reported Eph receptor structures 657:699 previously reported Eph receptor structures 657:699 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 64 theme other 770:774 arg1 receptors 788:796 other class A Eph receptors 770:796 other class A Eph receptors 770:796 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 1 65 dep kinase 102:107 arg1 receptor 122:129 cell surface receptor 109:129 The EphA4 tyrosine kinase cell surface receptor 83:129 The EphA4 tyrosine kinase cell surface receptor regulates an array of physiological processes and is the only currently known class A Eph receptor that binds both A and B class ephrins with high affinity. 19836338 6 66 theme B 975:975 arg1 conformations 996:1008 both A and B class Eph receptor conformations 964:1008 both A and B class Eph receptor conformations 964:1008 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 6 67 theme structural 928:937 arg1 EphA4 917:921 EphA4 917:921 EphA4 917:921 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 6 67 theme structural 928:937 arg1 chameleon 939:947 a structural chameleon 926:947 a structural chameleon 926:947 This interactive plasticity reveals EphA4 as a structural chameleon, able to adopt both A and B class Eph receptor conformations, and thus provides a molecular basis for EphA-type cross-class reactivity. 19836338 0 68 theme ephrin 65:70 arg1 signaling 72:80 cross-class ephrin signaling 53:80 cross-class ephrin signaling 53:80 Structural plasticity of eph receptor A4 facilitates cross-class ephrin signaling. 19836338 5 69 from EphA4 712:716 arg1 isolation 721:729 isolation 721:729 isolation 721:729 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 5 69 from EphA4 712:716 arg1 complex 738:744 complex 738:744 complex with ephrinA2 738:758 Structural analyses, drawing on previously reported Eph receptor structures, show that EphA4 in isolation and in complex with ephrinA2 resembles other class A Eph receptors but on binding ephrinB2 assumes structural hallmarks of the class B Eph receptors. 19836338 2 70 theme EphA4 332:336 arg1 domain 353:358 the EphA4 ligand binding domain 328:358 the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2 328:414 We have solved the crystal structure of the EphA4 ligand binding domain alone and in complex with (1) ephrinB2 and (2) ephrinA2. 29794134 0 0 theme R47H 87:90 arg1 variant 92:98 the Alzheimer's disease-associated R47H variant 52:98 the Alzheimer's disease-associated R47H variant of the immune receptor TREM2 52:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 2 1 theme loss 426:429 arg1 basis 406:410 the molecular basis 392:410 the molecular basis of R47H TREM2 loss of function 392:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 1 theme loss 426:429 arg1 area 458:461 an emerging area 446:461 an emerging area of TREM2 biology 446:478 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 6 2 theme structural 1173:1182 arg1 findings 1184:1191 our structural findings 1169:1191 our structural findings 1169:1191 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 6 3 dep in 1130:1131 arg1 vitro 1133:1137 vitro 1133:1137 vitro 1133:1137 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 2 4 theme TREM2 420:424 arg1 loss 426:429 R47H TREM2 loss 415:429 R47H TREM2 loss of function 415:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 6 5 theme in 1143:1144 arg1 characterization 1151:1166 in vitro and in vivo characterization 1130:1166 in vitro and in vivo characterization 1130:1166 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 1 6 theme myeloid 163:169 arg1 cells 171:175 myeloid cells 2 (TREM2) 163:185 myeloid cells 2 (TREM2) 163:185 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 7 7 theme function 1426:1433 arg1 loss 1418:1421 loss 1418:1421 loss of function in disease 1418:1444 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 4 8 theme complementarity-determining 786:812 arg1 loop 830:833 the complementarity-determining region 2 (CDR2) loop 782:833 the complementarity-determining region 2 (CDR2) loop 782:833 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 9 theme CDR2 824:827 arg1 loop 830:833 the complementarity-determining region 2 (CDR2) loop 782:833 the complementarity-determining region 2 (CDR2) loop 782:833 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 10 theme interaction 940:950 arg1 capable 922:928 capable 922:928 capable 922:928 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 1 11 theme microglia 237:245 arg1 surface 226:232 the surface 222:232 the surface of microglia, macrophages, dendritic cells, and osteoclasts 222:292 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 0 12 theme variant 92:98 arg1 effects 41:47 the loss-of-function effects 20:47 the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2 20:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 3 13 theme -bound 631:636 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 7 14 theme decreased 1354:1362 arg1 stability 1385:1393 decreased in vitro and in vivo stability 1354:1393 decreased in vitro and in vivo stability of TREM2 1354:1402 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 6 15 theme molecular 1207:1215 arg1 mechanism 1217:1225 the molecular mechanism 1203:1225 the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2 1203:1324 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 0 16 theme immune 107:112 arg1 TREM2 123:127 the immune receptor TREM2 103:127 the immune receptor TREM2 103:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 3 17 theme WT 638:639 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 2 18 theme biology 472:478 arg1 basis 406:410 the molecular basis 392:410 the molecular basis of R47H TREM2 loss of function 392:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 18 theme biology 472:478 arg1 factor 340:345 a significant risk factor 321:345 a significant risk factor for late-onset Alzheimer's disease (AD) 321:385 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 18 theme biology 472:478 arg1 variant 310:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 18 theme biology 472:478 arg1 area 458:461 an emerging area 446:461 an emerging area of TREM2 biology 446:478 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 6 19 theme binding 1253:1259 arg1 loss 1238:1241 loss 1238:1241 loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2 1238:1324 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 4 20 theme loop 830:833 arg1 PLIS 889:892 PLIS 889:892 PLIS 889:892 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 20 theme loop 830:833 arg1 surface 880:886 the putative positive ligand-interacting surface 839:886 the putative positive ligand-interacting surface (PLIS) 839:893 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 20 theme loop 830:833 arg1 features 770:777 the structural features 755:777 the structural features of the complementarity-determining region 2 (CDR2) loop 755:833 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 6 21 theme functional 1292:1301 arg1 activity 1303:1310 functional activity 1292:1310 functional activity 1292:1310 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 6 22 theme R47H 1315:1318 arg1 TREM2 1320:1324 R47H TREM2 1315:1324 R47H TREM2 1315:1324 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 5 23 theme PS-bound 980:987 arg1 structure 989:997 the PS-bound structure 976:997 the PS-bound structure 976:997 This is exemplified in the PS-bound structure, in which the CDR2 loop and PLIS drive critical interactions with PS via surfaces that are disrupted in the variant. 29794134 2 24 theme late-onset 351:360 arg1 disease 374:380 late-onset Alzheimer's disease 351:380 late-onset Alzheimer's disease (AD) 351:385 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 24 theme late-onset 351:360 arg1 AD 383:384 AD 383:384 AD 383:384 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 6 25 theme TREM2 1320:1324 arg1 binding 1253:1259 ligand binding 1246:1259 ligand binding 1246:1259 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 6 25 theme TREM2 1320:1324 arg1 oligomerization 1271:1285 putative oligomerization 1262:1285 putative oligomerization 1262:1285 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 6 25 theme TREM2 1320:1324 arg1 activity 1303:1310 functional activity 1292:1310 functional activity 1292:1310 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 0 26 theme disease-associated 68:85 arg1 variant 92:98 the Alzheimer's disease-associated R47H variant 52:98 the Alzheimer's disease-associated R47H variant of the immune receptor TREM2 52:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 4 27 theme critical 726:733 arg1 role 735:738 a critical role 724:738 a critical role 724:738 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 28 theme putative 843:850 arg1 PLIS 889:892 PLIS 889:892 PLIS 889:892 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 28 theme putative 843:850 arg1 surface 880:886 the putative positive ligand-interacting surface 839:886 the putative positive ligand-interacting surface (PLIS) 839:893 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 3 29 from domains 566:572 arg1 Å 668:668 1.8, 2.2, and 2.2 Å 650:668 1.8, 2.2, and 2.2 Å 650:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 0 30 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 0:128 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 1 31 theme macrophages 248:258 arg1 surface 226:232 the surface 222:232 the surface of microglia, macrophages, dendritic cells, and osteoclasts 222:292 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 7 32 theme in 1364:1365 arg1 stability 1385:1393 decreased in vitro and in vivo stability 1354:1393 decreased in vitro and in vivo stability of TREM2 1354:1402 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 6 33 theme in 1130:1131 arg1 characterization 1151:1166 in vitro and in vivo characterization 1130:1166 in vitro and in vivo characterization 1130:1166 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 4 34 theme capable 922:928 arg1 conformations 908:920 conformations 908:920 conformations capable of ligand interaction 908:950 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 7 35 dep in 1364:1365 arg1 vitro 1367:1371 vitro 1367:1371 vitro 1367:1371 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 0 36 theme TREM2 123:127 arg1 variant 92:98 the Alzheimer's disease-associated R47H variant 52:98 the Alzheimer's disease-associated R47H variant of the immune receptor TREM2 52:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 5 37 with interactions 1047:1058 arg1 PS 1065:1066 PS 1065:1066 PS 1065:1066 This is exemplified in the PS-bound structure, in which the CDR2 loop and PLIS drive critical interactions with PS via surfaces that are disrupted in the variant. 29794134 6 38 theme ligand 1246:1251 arg1 binding 1253:1259 ligand binding 1246:1259 ligand binding 1246:1259 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 6 39 theme putative 1262:1269 arg1 oligomerization 1271:1285 putative oligomerization 1262:1285 putative oligomerization 1262:1285 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 6 40 theme activity 1303:1310 arg1 loss 1238:1241 loss 1238:1241 loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2 1238:1324 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 1 41 theme dendritic 261:269 arg1 cells 271:275 dendritic cells 261:275 dendritic cells 261:275 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 0 42 theme receptor 114:121 arg1 TREM2 123:127 the immune receptor TREM2 103:127 the immune receptor TREM2 103:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 7 43 theme TREM2 1398:1402 arg1 stability 1385:1393 decreased in vitro and in vivo stability 1354:1393 decreased in vitro and in vivo stability of TREM2 1354:1402 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 2 44 theme R47H 415:418 arg1 loss 426:429 R47H TREM2 loss 415:429 R47H TREM2 loss of function 415:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 1 45 theme cells 271:275 arg1 surface 226:232 the surface 222:232 the surface of microglia, macrophages, dendritic cells, and osteoclasts 222:292 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 2 46 theme TREM2 466:470 arg1 biology 472:478 TREM2 biology 466:478 TREM2 biology 466:478 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 0 47 theme loss-of-function 24:39 arg1 effects 41:47 the loss-of-function effects 20:47 the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2 20:127 Molecular basis for the loss-of-function effects of the Alzheimer's disease-associated R47H variant of the immune receptor TREM2. 29794134 4 48 theme positive 852:859 arg1 PLIS 889:892 PLIS 889:892 PLIS 889:892 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 48 theme positive 852:859 arg1 surface 880:886 the putative positive ligand-interacting surface 839:886 the putative positive ligand-interacting surface (PLIS) 839:893 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 1 49 theme immune 193:198 arg1 receptor 200:207 an immune receptor 190:207 an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts 190:292 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 1 49 theme immune 193:198 arg1 receptor 141:148 Triggering receptor 130:148 Triggering receptor expressed on myeloid cells 2 (TREM2) 130:185 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 3 50 from Å 668:668 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 50 from Å 668:668 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 50 from Å 668:668 arg1 apo-WT 596:601 apo-WT 596:601 apo-WT 596:601 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 50 from Å 668:668 arg1 ECDs 575:578 ECDs 575:578 ECDs 575:578 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 50 from Å 668:668 arg1 domains 566:572 the extracellular ligand-binding domains 533:572 the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 533:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 1 51 dep cells 171:175 arg1 TREM2 180:184 TREM2 180:184 TREM2 180:184 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 1 51 dep cells 171:175 arg1 2 177:177 2 177:177 2 177:177 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 7 52 from loss 1418:1421 arg1 disease 1438:1444 disease 1438:1444 disease 1438:1444 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 2 53 theme molecular 396:404 arg1 basis 406:410 the molecular basis 392:410 the molecular basis of R47H TREM2 loss of function 392:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 53 theme molecular 396:404 arg1 area 458:461 an emerging area 446:461 an emerging area of TREM2 biology 446:478 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 1 54 theme osteoclasts 282:292 arg1 surface 226:232 the surface 222:232 the surface of microglia, macrophages, dendritic cells, and osteoclasts 222:292 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 4 55 theme region 814:819 arg1 loop 830:833 the complementarity-determining region 2 (CDR2) loop 782:833 the complementarity-determining region 2 (CDR2) loop 782:833 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 7 56 theme in 1377:1378 arg1 stability 1385:1393 decreased in vitro and in vivo stability 1354:1393 decreased in vitro and in vivo stability of TREM2 1354:1402 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 3 57 theme high-resolution 503:517 arg1 structures 519:528 three high-resolution structures 497:528 three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 497:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 2 58 theme TREM2 304:308 arg1 factor 340:345 a significant risk factor 321:345 a significant risk factor for late-onset Alzheimer's disease (AD) 321:385 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 58 theme TREM2 304:308 arg1 variant 310:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 4 59 theme structural 759:768 arg1 features 770:777 the structural features 755:777 the structural features of the complementarity-determining region 2 (CDR2) loop 755:833 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 3 60 theme apo-WT 596:601 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 60 theme apo-WT 596:601 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 60 theme apo-WT 596:601 arg1 apo-WT 596:601 apo-WT 596:601 apo-WT 596:601 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 60 theme apo-WT 596:601 arg1 ECDs 575:578 ECDs 575:578 ECDs 575:578 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 60 theme apo-WT 596:601 arg1 domains 566:572 the extracellular ligand-binding domains 533:572 the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 533:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 61 theme ligand-binding 551:564 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 61 theme ligand-binding 551:564 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 61 theme ligand-binding 551:564 arg1 apo-WT 596:601 apo-WT 596:601 apo-WT 596:601 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 61 theme ligand-binding 551:564 arg1 ECDs 575:578 ECDs 575:578 ECDs 575:578 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 61 theme ligand-binding 551:564 arg1 domains 566:572 the extracellular ligand-binding domains 533:572 the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 533:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 2 62 theme R47H 299:302 arg1 factor 340:345 a significant risk factor 321:345 a significant risk factor for late-onset Alzheimer's disease (AD) 321:385 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 62 theme R47H 299:302 arg1 variant 310:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 6 63 dep in 1143:1144 arg1 vivo 1146:1149 vivo 1146:1149 vivo 1146:1149 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 3 64 theme R47H 584:587 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 65 theme domains 566:572 arg1 structures 519:528 three high-resolution structures 497:528 three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 497:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 1 66 theme Triggering 130:139 arg1 receptor 200:207 an immune receptor 190:207 an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts 190:292 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 1 66 theme Triggering 130:139 arg1 receptor 141:148 Triggering receptor 130:148 Triggering receptor expressed on myeloid cells 2 (TREM2) 130:185 Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on the surface of microglia, macrophages, dendritic cells, and osteoclasts. 29794134 3 67 theme TREM2 641:645 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 67 theme TREM2 641:645 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 67 theme TREM2 641:645 arg1 apo-WT 596:601 apo-WT 596:601 apo-WT 596:601 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 67 theme TREM2 641:645 arg1 ECDs 575:578 ECDs 575:578 ECDs 575:578 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 67 theme TREM2 641:645 arg1 domains 566:572 the extracellular ligand-binding domains 533:572 the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 533:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 2 68 theme function 434:441 arg1 loss 426:429 R47H TREM2 loss 415:429 R47H TREM2 loss of function 415:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 3 69 theme TREM2 589:593 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 69 theme TREM2 589:593 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 69 theme TREM2 589:593 arg1 apo-WT 596:601 apo-WT 596:601 apo-WT 596:601 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 69 theme TREM2 589:593 arg1 ECDs 575:578 ECDs 575:578 ECDs 575:578 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 69 theme TREM2 589:593 arg1 domains 566:572 the extracellular ligand-binding domains 533:572 the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 533:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 5 70 theme CDR2 1013:1016 arg1 loop 1018:1021 the CDR2 loop 1009:1021 the CDR2 loop 1009:1021 This is exemplified in the PS-bound structure, in which the CDR2 loop and PLIS drive critical interactions with PS via surfaces that are disrupted in the variant. 29794134 2 71 theme risk 335:338 arg1 factor 340:345 a significant risk factor 321:345 a significant risk factor for late-onset Alzheimer's disease (AD) 321:385 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 71 theme risk 335:338 arg1 variant 310:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 5 72 theme critical 1038:1045 arg1 interactions 1047:1058 critical interactions 1038:1058 critical interactions with PS 1038:1066 This is exemplified in the PS-bound structure, in which the CDR2 loop and PLIS drive critical interactions with PS via surfaces that are disrupted in the variant. 29794134 7 73 dep in 1377:1378 arg1 vivo 1380:1383 vivo 1380:1383 vivo 1380:1383 They also help unravel how decreased in vitro and in vivo stability of TREM2 contribute to loss of function in disease. 29794134 4 74 theme ligand-interacting 861:878 arg1 PLIS 889:892 PLIS 889:892 PLIS 889:892 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 4 74 theme ligand-interacting 861:878 arg1 surface 880:886 the putative positive ligand-interacting surface 839:886 the putative positive ligand-interacting surface (PLIS) 839:893 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 3 75 theme extracellular 537:549 arg1 TREM2 589:593 R47H TREM2 584:593 R47H TREM2 584:593 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 75 theme extracellular 537:549 arg1 TREM2 641:645 phosphatidylserine (PS)-bound WT TREM2 608:645 phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 608:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 75 theme extracellular 537:549 arg1 apo-WT 596:601 apo-WT 596:601 apo-WT 596:601 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 75 theme extracellular 537:549 arg1 ECDs 575:578 ECDs 575:578 ECDs 575:578 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 3 75 theme extracellular 537:549 arg1 domains 566:572 the extracellular ligand-binding domains 533:572 the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å 533:668 Here, we report three high-resolution structures of the extracellular ligand-binding domains (ECDs) of R47H TREM2, apo-WT, and phosphatidylserine (PS)-bound WT TREM2 at 1.8, 2.2, and 2.2 Å, respectively. 29794134 6 76 theme oligomerization 1271:1285 arg1 loss 1238:1241 loss 1238:1241 loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2 1238:1324 Together with in vitro and in vivo characterization, our structural findings elucidate the molecular mechanism underlying loss of ligand binding, putative oligomerization, and functional activity of R47H TREM2. 29794134 2 77 theme significant 323:333 arg1 factor 340:345 a significant risk factor 321:345 a significant risk factor for late-onset Alzheimer's disease (AD) 321:385 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 77 theme significant 323:333 arg1 variant 310:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant 295:316 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 4 78 theme ligand 933:938 arg1 interaction 940:950 ligand interaction 933:950 ligand interaction 933:950 The structures reveal that Arg47 plays a critical role in maintaining the structural features of the complementarity-determining region 2 (CDR2) loop and the putative positive ligand-interacting surface (PLIS), stabilizing conformations capable of ligand interaction. 29794134 2 79 theme emerging 449:456 arg1 basis 406:410 the molecular basis 392:410 the molecular basis of R47H TREM2 loss of function 392:441 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 29794134 2 79 theme emerging 449:456 arg1 area 458:461 an emerging area 446:461 an emerging area of TREM2 biology 446:478 The R47H TREM2 variant is a significant risk factor for late-onset Alzheimer's disease (AD), and the molecular basis of R47H TREM2 loss of function is an emerging area of TREM2 biology. 32767432 1 0 theme cell 167:170 arg1 surface 172:178 the cell surface 163:178 the cell surface 163:178 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 1 theme purinergic 249:258 arg1 signaling 260:268 purinergic signaling 249:268 purinergic signaling 249:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 2 theme ecto-nucleoside 70:84 arg1 NTPDases 120:127 NTPDases 120:127 NTPDases 120:127 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 2 theme ecto-nucleoside 70:84 arg1 diphosphohydrolases 99:117 The ecto-nucleoside triphosphate diphosphohydrolases 66:117 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) 66:128 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 2 theme ecto-nucleoside 70:84 arg1 family 136:141 a family 134:141 a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling 134:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 2 3 theme intracellular 277:289 arg1 roles 291:295 Their intracellular roles 271:295 Their intracellular roles 271:295 Their intracellular roles, however, have not been clearly defined. 32767432 1 4 theme signaling 260:268 arg1 enzymes 146:152 enzymes 146:152 enzymes 146:152 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 4 theme signaling 260:268 arg1 regulators 235:244 major regulators 229:244 major regulators of purinergic signaling 229:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 5 located found 154:158 arg1 surface 172:178 the cell surface 163:178 the cell surface 163:178 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 5 located found 154:158 arg2 enzymes 146:152 enzymes 146:152 enzymes 146:152 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 5 located found 154:158 arg2 regulators 235:244 major regulators 229:244 major regulators of purinergic signaling 229:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 5 located found 154:158 arg1 lumen 191:195 the lumen 187:195 the lumen of certain organelles 187:217 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 6 6 theme substrate 756:764 arg1 selectivity 766:776 its substrate selectivity 752:776 its substrate selectivity 752:776 Differences in the nucleotide-binding site relative to its homologs could account for its substrate selectivity. 32767432 1 7 theme triphosphate 86:97 arg1 NTPDases 120:127 NTPDases 120:127 NTPDases 120:127 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 7 theme triphosphate 86:97 arg1 diphosphohydrolases 99:117 The ecto-nucleoside triphosphate diphosphohydrolases 66:117 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) 66:128 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 7 theme triphosphate 86:97 arg1 family 136:141 a family 134:141 a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling 134:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 7 8 theme NTPDases 834:841 arg1 loop 813:816 The putative membrane-interacting loop 779:816 The putative membrane-interacting loop of cell-surface NTPDases 779:841 The putative membrane-interacting loop of cell-surface NTPDases is drastically altered in NTPDase4, potentially affecting its interdomain dynamics at the Golgi membrane. 32767432 3 9 theme nucleotide 407:416 arg1 recycling 418:426 nucleotide recycling 407:426 nucleotide recycling 407:426 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 7 10 theme cell-surface 821:832 arg1 NTPDases 834:841 cell-surface NTPDases 821:841 cell-surface NTPDases 821:841 The putative membrane-interacting loop of cell-surface NTPDases is drastically altered in NTPDase4, potentially affecting its interdomain dynamics at the Golgi membrane. 32767432 7 11 theme Golgi 933:937 arg1 membrane 939:946 the Golgi membrane 929:946 the Golgi membrane 929:946 The putative membrane-interacting loop of cell-surface NTPDases is drastically altered in NTPDase4, potentially affecting its interdomain dynamics at the Golgi membrane. 32767432 5 12 theme wide 632:635 arg1 conformation 652:663 a wide open, inactive conformation 630:663 conformation 652:663 The enzyme adopts a wide open, inactive conformation. 32767432 0 13 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the nucleotide-metabolizing enzyme 0:54 Crystal structure of the nucleotide-metabolizing enzyme NTPDase4. 32767432 5 14 theme inactive 643:650 arg1 conformation 652:663 a wide open, inactive conformation 630:663 conformation 652:663 The enzyme adopts a wide open, inactive conformation. 32767432 6 15 from Differences 666:676 arg1 site 704:707 the nucleotide-binding site 681:707 the nucleotide-binding site relative to its homologs 681:732 Differences in the nucleotide-binding site relative to its homologs could account for its substrate selectivity. 32767432 7 16 theme putative 783:790 arg1 loop 813:816 The putative membrane-interacting loop 779:816 The putative membrane-interacting loop of cell-surface NTPDases 779:841 The putative membrane-interacting loop of cell-surface NTPDases is drastically altered in NTPDase4, potentially affecting its interdomain dynamics at the Golgi membrane. 32767432 3 17 theme protein 439:445 arg1 glycosylation 447:459 protein glycosylation 439:459 protein glycosylation 439:459 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 0 18 theme enzyme 49:54 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the nucleotide-metabolizing enzyme 0:54 Crystal structure of the nucleotide-metabolizing enzyme NTPDase4. 32767432 7 19 theme membrane-interacting 792:811 arg1 loop 813:816 The putative membrane-interacting loop 779:816 The putative membrane-interacting loop of cell-surface NTPDases 779:841 The putative membrane-interacting loop of cell-surface NTPDases is drastically altered in NTPDase4, potentially affecting its interdomain dynamics at the Golgi membrane. 32767432 1 20 theme certain 200:206 arg1 organelles 208:217 certain organelles 200:217 certain organelles 200:217 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 0 21 theme nucleotide-metabolizing 25:47 arg1 enzyme 49:54 the nucleotide-metabolizing enzyme 21:54 the nucleotide-metabolizing enzyme 21:54 Crystal structure of the nucleotide-metabolizing enzyme NTPDase4. 32767432 4 22 theme crystal 593:599 arg1 structure 601:609 its crystal structure 589:609 its crystal structure 589:609 To further our understanding of NTPDase4 function, we determined its crystal structure. 32767432 3 23 theme Golgi 369:373 arg1 NTPDase4 338:345 NTPDase4 338:345 NTPDase4 (UDPase, ENTPD4) 338:362 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 3 23 theme Golgi 369:373 arg1 protein 375:381 a Golgi protein 367:381 a Golgi protein potentially involved in nucleotide recycling 367:426 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 3 24 dep NTPDase4 338:345 arg1 ENTPD4 356:361 ENTPD4 356:361 ENTPD4 356:361 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 3 24 dep NTPDase4 338:345 arg1 UDPase 348:353 UDPase 348:353 UDPase 348:353 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 3 25 located found 474:478 arg2 NTPDase4 338:345 NTPDase4 338:345 NTPDase4 (UDPase, ENTPD4) 338:362 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 3 25 located found 474:478 arg2 protein 375:381 a Golgi protein 367:381 a Golgi protein potentially involved in nucleotide recycling 367:426 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 3 25 located found 474:478 arg1 lysosomes 483:491 lysosomes 483:491 lysosomes 483:491 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 5 26 theme open 637:640 arg1 conformation 652:663 a wide open, inactive conformation 630:663 conformation 652:663 The enzyme adopts a wide open, inactive conformation. 32767432 6 27 theme relative 709:716 arg1 site 704:707 the nucleotide-binding site 681:707 the nucleotide-binding site relative to its homologs 681:732 Differences in the nucleotide-binding site relative to its homologs could account for its substrate selectivity. 32767432 3 28 theme glycosylation 447:459 arg1 part 431:434 part 431:434 part of protein glycosylation 431:459 NTPDase4 (UDPase, ENTPD4) is a Golgi protein potentially involved in nucleotide recycling as part of protein glycosylation, and is also found in lysosomes, where its purpose is unknown. 32767432 7 29 theme interdomain 905:915 arg1 dynamics 917:924 its interdomain dynamics 901:924 its interdomain dynamics 901:924 The putative membrane-interacting loop of cell-surface NTPDases is drastically altered in NTPDase4, potentially affecting its interdomain dynamics at the Golgi membrane. 32767432 4 30 theme function 565:572 arg1 understanding 539:551 our understanding 535:551 our understanding of NTPDase4 function 535:572 To further our understanding of NTPDase4 function, we determined its crystal structure. 32767432 1 31 theme enzymes 146:152 arg1 diphosphohydrolases 99:117 The ecto-nucleoside triphosphate diphosphohydrolases 66:117 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) 66:128 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 31 theme enzymes 146:152 arg1 family 136:141 a family 134:141 a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling 134:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 4 32 theme NTPDase4 556:563 arg1 function 565:572 NTPDase4 function 556:572 NTPDase4 function 556:572 To further our understanding of NTPDase4 function, we determined its crystal structure. 32767432 1 33 theme organelles 208:217 arg1 lumen 191:195 the lumen 187:195 the lumen of certain organelles 187:217 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 6 34 theme nucleotide-binding 685:702 arg1 site 704:707 the nucleotide-binding site 681:707 the nucleotide-binding site relative to its homologs 681:732 Differences in the nucleotide-binding site relative to its homologs could account for its substrate selectivity. 32767432 1 35 theme major 229:233 arg1 enzymes 146:152 enzymes 146:152 enzymes 146:152 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 32767432 1 35 theme major 229:233 arg1 regulators 235:244 major regulators 229:244 major regulators of purinergic signaling 229:268 The ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are a family of enzymes found on the cell surface and in the lumen of certain organelles, that are major regulators of purinergic signaling. 20944746 10 0 theme developmental 2000:2012 arg1 control 2022:2028 developmental quality control 2000:2028 developmental quality control 2000:2028 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 3 1 theme TCR 664:666 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 9 2 theme β-chain 1781:1787 arg1 V 1754:1754 both the V and C domains 1745:1768 V 1754:1754 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 9 2 theme β-chain 1781:1787 arg1 domains 1762:1768 both the V and C domains 1745:1768 domains 1762:1768 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 9 2 theme β-chain 1781:1787 arg1 β-chain 1781:1787 any TCR β-chain 1773:1787 any TCR β-chain 1773:1787 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 6 3 theme pre-TCR 1116:1122 arg1 dimer 1124:1128 the pre-TCR dimer 1112:1128 the pre-TCR dimer 1112:1128 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 7 4 theme β 1368:1368 arg1 families 1375:1382 β gene families 1368:1382 β gene families 1368:1382 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 1 5 theme pivotal 168:174 arg1 role 176:179 a pivotal role 166:179 a pivotal role 166:179 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 6 6 theme molecular 1090:1098 arg1 envelope 1100:1107 the molecular envelope 1086:1107 the molecular envelope of the pre-TCR dimer 1086:1128 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 10 7 theme membrane 2085:2092 arg1 complex 2103:2109 an integral membrane receptor complex 2073:2109 an integral membrane receptor complex 2073:2109 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 1 8 theme T-cell 326:331 arg1 differentiation 341:355 subsequent αβ T-cell lineage differentiation 312:355 subsequent αβ T-cell lineage differentiation 312:355 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 2 9 theme dimerization 532:543 arg1 event 545:549 a ligand-independent dimerization event 511:549 a ligand-independent dimerization event 511:549 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 1 10 theme T-cell 190:195 arg1 development 197:207 early T-cell development 184:207 early T-cell development 184:207 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 6 11 theme pre-TCR 1177:1183 arg1 dimer 1185:1189 the observed head-to-tail pre-TCR dimer 1151:1189 the observed head-to-tail pre-TCR dimer 1151:1189 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 10 12 theme receptor 2094:2101 arg1 complex 2103:2109 an integral membrane receptor complex 2073:2109 an integral membrane receptor complex 2073:2109 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 5 13 theme TCR 895:897 arg1 α-chain 899:905 the TCR α-chain 891:905 the TCR α-chain 891:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 9 14 theme pre-TCR 1641:1647 arg1 self-association 1649:1664 pre-TCR self-association 1641:1664 pre-TCR self-association 1641:1664 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 6 15 theme observed 1155:1162 arg1 dimer 1185:1189 the observed head-to-tail pre-TCR dimer 1151:1189 the observed head-to-tail pre-TCR dimer 1151:1189 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 10 16 theme integral 2076:2083 arg1 complex 2103:2109 an integral membrane receptor complex 2073:2109 an integral membrane receptor complex 2073:2109 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 10 17 theme sensing 1925:1931 arg1 function 1933:1940 This unusual dual-chaperone-like sensing function 1892:1940 This unusual dual-chaperone-like sensing function of pre-Tα 1892:1950 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 9 18 theme T-cell 1872:1877 arg1 development 1879:1889 T-cell development 1872:1889 T-cell development 1872:1889 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 10 19 theme unusual 1897:1903 arg1 function 1933:1940 This unusual dual-chaperone-like sensing function 1892:1940 This unusual dual-chaperone-like sensing function of pre-Tα 1892:1950 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 0 20 theme pre-T-cell 56:65 arg1 receptor 75:82 the pre-T-cell antigen receptor 52:82 the pre-T-cell antigen receptor 52:82 The structural basis for autonomous dimerization of the pre-T-cell antigen receptor. 20944746 2 21 theme pre-TCR-induced 465:479 arg1 signalling 481:490 pre-TCR-induced signalling 465:490 pre-TCR-induced signalling 465:490 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 0 22 theme receptor 75:82 arg1 dimerization 36:47 autonomous dimerization 25:47 autonomous dimerization of the pre-T-cell antigen receptor 25:82 The structural basis for autonomous dimerization of the pre-T-cell antigen receptor. 20944746 1 23 theme double-negative 280:294 arg1 thymocytes 296:305 CD4(-)CD8(-) double-negative thymocytes 267:305 CD4(-)CD8(-) double-negative thymocytes 267:305 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 5 24 from domain 881:886 arg1 distinct 850:857 distinct 850:857 distinct 850:857 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 24 from domain 881:886 arg1 domain 822:827 a single immunoglobulin-like domain 793:827 a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain 793:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 7 25 dep Vβ 1349:1350 arg1 the 1345:1347 the 1345:1347 the 1345:1347 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 7 25 dep Vβ 1349:1350 arg1 families 1375:1382 β gene families 1368:1382 β gene families 1368:1382 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 6 26 contain had 1040:1042 arg1 pre-TCR 1032:1038 The pre-TCR 1028:1038 The pre-TCR 1028:1038 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 6 26 contain had 1040:1042 arg2 propensity 1046:1055 a propensity 1044:1055 a propensity to dimerize in solution 1044:1079 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 4 27 theme pre-TCR 744:750 arg1 dimerization 752:763 pre-TCR dimerization 744:763 pre-TCR dimerization 744:763 Here we provide the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization. 20944746 5 28 theme association 934:944 arg1 mode 926:929 the mode 922:929 the mode of association between pre-Tα and TCRβ 922:968 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 7 29 theme interface 1450:1458 arg1 core 1424:1427 the core 1420:1427 the core of the αβTCR's Vα-Vβ interface 1420:1458 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 1 30 theme subsequent 312:321 arg1 differentiation 341:355 subsequent αβ T-cell lineage differentiation 312:355 subsequent αβ T-cell lineage differentiation 312:355 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 7 31 theme V 1283:1283 arg1 domain 1288:1293 the variable (V) β domain 1269:1293 the variable (V) β domain 1269:1293 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 2 32 theme histocompatibility 409:426 arg1 complex 428:434 the peptide-loaded major histocompatibility complex 384:434 the peptide-loaded major histocompatibility complex 384:434 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 5 33 theme single 795:800 arg1 distinct 850:857 distinct 850:857 distinct 850:857 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 33 theme single 795:800 arg1 domain 822:827 a single immunoglobulin-like domain 793:827 a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain 793:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 1 34 theme pre-T-cell 89:98 arg1 pre-TCR 118:124 pre-TCR 118:124 pre-TCR 118:124 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 34 theme pre-T-cell 89:98 arg1 receptor 108:115 The pre-T-cell antigen receptor 85:115 The pre-T-cell antigen receptor (pre-TCR) 85:125 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 7 35 theme variable 1273:1280 arg1 domain 1288:1293 the variable (V) β domain 1269:1293 the variable (V) β domain 1269:1293 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 2 36 theme peptide-loaded 388:401 arg1 complex 428:434 the peptide-loaded major histocompatibility complex 384:434 the peptide-loaded major histocompatibility complex 384:434 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 4 37 theme assembly 731:738 arg1 basis 710:714 the basis 706:714 the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization 706:763 Here we provide the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization. 20944746 1 38 theme CD4 267:269 arg1 thymocytes 296:305 CD4(-)CD8(-) double-negative thymocytes 267:305 CD4(-)CD8(-) double-negative thymocytes 267:305 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 9 39 from checkpoint 1858:1867 arg1 development 1879:1889 T-cell development 1872:1889 T-cell development 1872:1889 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 2 40 theme αβTCR 366:370 arg1 ligation 372:379 αβTCR ligation 366:379 αβTCR ligation by the peptide-loaded major histocompatibility complex 366:434 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 5 41 theme pre-Tα 770:775 arg1 chain 777:781 The pre-Tα chain 766:781 The pre-Tα chain 766:781 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 0 42 theme autonomous 25:34 arg1 dimerization 36:47 autonomous dimerization 25:47 autonomous dimerization of the pre-T-cell antigen receptor 25:82 The structural basis for autonomous dimerization of the pre-T-cell antigen receptor. 20944746 3 43 theme TCR 624:626 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 1 44 theme CD8 273:275 arg1 thymocytes 296:305 CD4(-)CD8(-) double-negative thymocytes 267:305 CD4(-)CD8(-) double-negative thymocytes 267:305 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 5 45 theme Cα-Cβ 1000:1004 arg1 domains 1006:1012 the Cα-Cβ domains 996:1012 the Cα-Cβ domains of the αβTCR 996:1025 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 45 theme Cα-Cβ 1000:1004 arg1 αβTCR 1021:1025 the αβTCR 1017:1025 the αβTCR 1017:1025 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 7 46 theme pre-Tα 1238:1243 arg1 domain 1245:1250 the pre-Tα domain 1234:1250 the pre-Tα domain 1234:1250 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 9 47 theme V 1754:1754 arg1 sample 1714:1719 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development 1713:1889 sample 1714:1719 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 8 48 theme pre-Tα-Vβ 1480:1488 arg1 interface 1496:1504 this pre-Tα-Vβ dimer interface 1475:1504 this pre-Tα-Vβ dimer interface 1475:1504 Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. 20944746 3 49 theme successful 653:662 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 8 50 theme pre-TCR 1562:1568 arg1 expression 1570:1579 pre-TCR expression 1562:1579 pre-TCR expression 1562:1579 Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. 20944746 9 51 theme C 1760:1760 arg1 domains 1762:1768 both the V and C domains 1745:1768 domains 1762:1768 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 9 51 theme C 1760:1760 arg1 β-chain 1781:1787 any TCR β-chain 1773:1787 any TCR β-chain 1773:1787 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 8 52 theme interface 1496:1504 arg1 Disruption 1461:1470 Disruption 1461:1470 Disruption of this pre-Tα-Vβ dimer interface 1461:1504 Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. 20944746 10 53 dep expression 2044:2053 arg1 the 2040:2042 the 2040:2042 the 2040:2042 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 7 54 theme pre-TCR 1205:1211 arg1 dimerization 1213:1224 pre-TCR dimerization 1205:1224 pre-TCR dimerization 1205:1224 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 10 55 theme quality 2014:2020 arg1 control 2022:2028 developmental quality control 2000:2028 developmental quality control 2000:2028 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 8 56 theme pre-TCR 1516:1522 arg1 dimerization 1524:1535 pre-TCR dimerization 1516:1535 pre-TCR dimerization 1516:1535 Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. 20944746 8 57 theme cell 1588:1591 arg1 surface 1593:1599 the cell surface 1584:1599 the cell surface 1584:1599 Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. 20944746 9 58 theme TCR 1777:1779 arg1 β-chain 1781:1787 any TCR β-chain 1773:1787 any TCR β-chain 1773:1787 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 3 59 theme β-gene 668:673 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 9 60 dep sample 1714:1719 arg1 folding 1734:1740 the correct folding 1722:1740 the correct folding 1722:1740 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 7 61 theme gene 1370:1373 arg1 families 1375:1382 β gene families 1368:1382 β gene families 1368:1382 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 6 62 theme dimer 1124:1128 arg1 envelope 1100:1107 the molecular envelope 1086:1107 the molecular envelope of the pre-TCR dimer 1086:1128 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 5 63 theme constant 868:875 arg1 domain 881:886 the constant (C) domain 864:886 the constant (C) domain of the TCR α-chain 864:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 63 theme constant 868:875 arg1 α-chain 899:905 the TCR α-chain 891:905 the TCR α-chain 891:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 9 64 theme ultimate 1808:1815 arg1 specificity 1817:1827 its ultimate specificity 1804:1827 its ultimate specificity 1804:1827 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 1 65 theme lineage 333:339 arg1 differentiation 341:355 subsequent αβ T-cell lineage differentiation 312:355 subsequent αβ T-cell lineage differentiation 312:355 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 66 theme early 184:188 arg1 development 197:207 early T-cell development 184:207 early T-cell development 184:207 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 2 67 theme ligand-independent 513:530 arg1 event 545:549 a ligand-independent dimerization event 511:549 a ligand-independent dimerization event 511:549 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 10 68 theme complex 2103:2109 arg1 signalling 2059:2068 signalling 2059:2068 signalling 2059:2068 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 10 68 theme complex 2103:2109 arg1 expression 2044:2053 expression 2044:2053 expression 2044:2053 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 3 69 theme invariant 577:585 arg1 pre-Tα 596:601 pre-Tα 596:601 pre-Tα 596:601 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 3 69 theme invariant 577:585 arg1 α-chain 587:593 an invariant α-chain 574:593 an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 574:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 6 70 theme head-to-tail 1164:1175 arg1 dimer 1185:1189 the observed head-to-tail pre-TCR dimer 1151:1189 the observed head-to-tail pre-TCR dimer 1151:1189 The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. 20944746 9 71 theme critical 1849:1856 arg1 checkpoint 1858:1867 a critical checkpoint 1847:1867 a critical checkpoint in T-cell development 1847:1889 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 5 72 theme α-chain 899:905 arg1 domain 881:886 the constant (C) domain 864:886 the constant (C) domain of the TCR α-chain 864:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 72 theme α-chain 899:905 arg1 α-chain 899:905 the TCR α-chain 891:905 the TCR α-chain 891:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 9 73 theme self-association 1649:1664 arg1 mechanism 1628:1636 a mechanism 1626:1636 a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development 1626:1889 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 10 74 theme dual-chaperone-like 1905:1923 arg1 function 1933:1940 This unusual dual-chaperone-like sensing function 1892:1940 This unusual dual-chaperone-like sensing function of pre-Tα 1892:1950 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 0 75 theme antigen 67:73 arg1 receptor 75:82 the pre-T-cell antigen receptor 52:82 the pre-T-cell antigen receptor 52:82 The structural basis for autonomous dimerization of the pre-T-cell antigen receptor. 20944746 5 76 theme αβTCR 1021:1025 arg1 domains 1006:1012 the Cα-Cβ domains 996:1012 the Cα-Cβ domains of the αβTCR 996:1025 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 76 theme αβTCR 1021:1025 arg1 αβTCR 1021:1025 the αβTCR 1017:1025 the αβTCR 1017:1025 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 77 theme C 878:878 arg1 domain 881:886 the constant (C) domain 864:886 the constant (C) domain of the TCR α-chain 864:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 77 theme C 878:878 arg1 α-chain 899:905 the TCR α-chain 891:905 the TCR α-chain 891:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 7 78 theme β 1286:1286 arg1 domain 1288:1293 the variable (V) β domain 1269:1293 the variable (V) β domain 1269:1293 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 3 79 with pairs 609:613 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 1 80 theme immature 141:148 arg1 thymocytes 150:159 immature thymocytes 141:159 immature thymocytes 141:159 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 9 81 theme pre-Tα 1682:1687 arg1 chain 1689:1693 the pre-Tα chain 1678:1693 the pre-Tα chain 1678:1693 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 10 82 theme pre-Tα 1945:1950 arg1 function 1933:1940 This unusual dual-chaperone-like sensing function 1892:1940 This unusual dual-chaperone-like sensing function of pre-Tα 1892:1950 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 1 83 theme thymocytes 296:305 arg1 differentiation 341:355 subsequent αβ T-cell lineage differentiation 312:355 subsequent αβ T-cell lineage differentiation 312:355 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 83 theme thymocytes 296:305 arg1 proliferation 250:262 proliferation 250:262 proliferation 250:262 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 83 theme thymocytes 296:305 arg1 survival 237:244 survival 237:244 survival 237:244 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 83 theme thymocytes 296:305 arg1 β-selection 224:234 TCR β-selection 220:234 TCR β-selection 220:234 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 7 84 theme Vα-Vβ 1444:1448 arg1 interface 1450:1458 the αβTCR's Vα-Vβ interface 1432:1458 the αβTCR's Vα-Vβ interface 1432:1458 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 10 85 theme unique 1965:1970 arg1 mechanism 1972:1980 a unique mechanism 1963:1980 a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex 1963:2109 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 4 86 theme dimerization 752:763 arg1 basis 710:714 the basis 706:714 the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization 706:763 Here we provide the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization. 20944746 0 87 theme structural 4:13 arg1 basis 15:19 The structural basis 0:19 The structural basis for autonomous dimerization of the pre-T-cell antigen receptor 0:82 The structural basis for autonomous dimerization of the pre-T-cell antigen receptor. 20944746 1 88 theme αβ 323:324 arg1 differentiation 341:355 subsequent αβ T-cell lineage differentiation 312:355 subsequent αβ T-cell lineage differentiation 312:355 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 2 89 theme major 403:407 arg1 complex 428:434 the peptide-loaded major histocompatibility complex 384:434 the peptide-loaded major histocompatibility complex 384:434 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 5 90 theme immunoglobulin-like 802:820 arg1 distinct 850:857 distinct 850:857 distinct 850:857 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 5 90 theme immunoglobulin-like 802:820 arg1 domain 822:827 a single immunoglobulin-like domain 793:827 a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain 793:905 The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. 20944746 1 91 theme antigen 100:106 arg1 pre-TCR 118:124 pre-TCR 118:124 pre-TCR 118:124 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 91 theme antigen 100:106 arg1 receptor 108:115 The pre-T-cell antigen receptor 85:115 The pre-T-cell antigen receptor (pre-TCR) 85:125 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 9 92 theme correct 1726:1732 arg1 folding 1734:1740 the correct folding 1722:1740 the correct folding 1722:1740 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 3 93 theme β-chain 628:634 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 4 94 theme pre-Tα-TCRβ 719:729 arg1 assembly 731:738 pre-Tα-TCRβ assembly 719:738 pre-Tα-TCRβ assembly 719:738 Here we provide the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization. 20944746 2 95 theme T-cell 446:451 arg1 signalling 453:462 T-cell signalling 446:462 T-cell signalling 446:462 Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. 20944746 1 96 contain has 162:164 arg1 pre-TCR 118:124 pre-TCR 118:124 pre-TCR 118:124 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 96 contain has 162:164 arg2 role 176:179 a pivotal role 166:179 a pivotal role 166:179 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 1 96 contain has 162:164 arg1 receptor 108:115 The pre-T-cell antigen receptor 85:115 The pre-T-cell antigen receptor (pre-TCR) 85:125 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20944746 8 97 theme dimer 1490:1494 arg1 interface 1496:1504 this pre-Tα-Vβ dimer interface 1475:1504 this pre-Tα-Vβ dimer interface 1475:1504 Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. 20944746 3 98 theme following 643:651 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 7 99 theme dimerization 1213:1224 arg1 mode 1197:1200 This mode 1192:1200 This mode of pre-TCR dimerization 1192:1224 This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. 20944746 10 100 from mechanism 1972:1980 arg1 nature 1985:1990 nature 1985:1990 nature 1985:1990 This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. 20944746 3 101 theme TCRβ 637:640 arg1 rearrangement 675:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement 620:687 The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. 20944746 9 102 theme domains 1762:1768 arg1 sample 1714:1719 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development 1713:1889 sample 1714:1719 Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. 20944746 1 103 theme TCR 220:222 arg1 β-selection 224:234 TCR β-selection 220:234 TCR β-selection 220:234 The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4(-)CD8(-) double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. 20934432 6 0 theme inter-domain 833:844 arg1 region 852:857 inter-domain hinge region 833:857 inter-domain hinge region 833:857 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 7 1 theme interface 907:915 arg1 architecture 887:898 architecture 887:898 architecture 887:898 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 7 1 theme interface 907:915 arg1 Composition 871:881 Composition 871:881 Composition 871:881 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 1 2 theme IFNs 209:212 arg1 group 191:195 the recently discovered group 167:195 the recently discovered group of type III IFNs 167:212 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 1 2 theme IFNs 209:212 arg1 IFNs 209:212 type III IFNs 200:212 type III IFNs 200:212 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 0 3 theme receptor 75:82 arg1 interferon-λR1 84:97 its high-affinity receptor interferon-λR1 57:97 its high-affinity receptor interferon-λR1 57:97 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 0 4 from structure 8:16 arg1 complex 44:50 complex 44:50 complex with its high-affinity receptor interferon-λR1 44:97 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 2 5 theme signaling 242:250 arg1 processes 252:260 signaling processes 242:260 signaling processes 242:260 All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2. 20934432 6 6 dep helix 728:732 arg1 AB 742:743 loop AB 737:743 loop AB 737:743 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 6 dep helix 728:732 arg1 F 756:756 helix F 750:756 helix F 750:756 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 6 dep helix 728:732 arg1 A 734:734 A 734:734 A 734:734 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 4 7 theme IL-10 552:556 arg1 members 568:574 other members 562:574 other members of the IL-10 family of cytokines 562:607 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 4 7 theme IL-10 552:556 arg1 structure 539:547 the structure 535:547 the structure of IL-10 535:556 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 6 8 theme N-terminal 811:820 arg1 IFN-λR1 862:868 IFN-λR1 862:868 IFN-λR1 862:868 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 8 theme N-terminal 811:820 arg1 domain 822:827 the N-terminal domain 807:827 the N-terminal domain 807:827 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 7 9 theme direct 942:947 arg1 bonds 958:962 only a few direct hydrogen bonds 931:962 only a few direct hydrogen bonds 931:962 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 6 10 theme helix 750:754 arg1 F 756:756 helix F 750:756 helix F 750:756 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 3 11 theme unique 450:455 arg1 IFN-λR1 430:436 human IFN-λR1 424:436 human IFN-λR1 424:436 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 3 11 theme unique 450:455 arg1 receptor 441:448 a receptor 439:448 a receptor unique to type III IFNs 439:472 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 7 12 theme ionic 996:1000 arg1 interactions 1002:1013 long-range ionic interactions 985:1013 long-range ionic interactions between ligand and receptor 985:1041 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 6 13 from helix 728:732 arg1 site 769:772 the IFN site 761:772 the IFN site 761:772 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 4 14 theme cytokines 599:607 arg1 family 589:594 the IL-10 family 579:594 the IL-10 family of cytokines 579:607 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 5 15 contain having 651:656 arg2 topology 679:686 fibronectin type III topology 658:686 fibronectin type III topology 658:686 IFN-λR1 consists of two distinct domains having fibronectin type III topology. 20934432 5 15 contain having 651:656 arg1 domains 643:649 two distinct domains 630:649 two distinct domains having fibronectin type III topology 630:686 IFN-λR1 consists of two distinct domains having fibronectin type III topology. 20934432 3 16 theme human 396:400 arg1 IFN-λ1 402:407 human IFN-λ1 396:407 human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs 396:472 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 0 17 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 0:98 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 6 18 theme loop 737:740 arg1 AB 742:743 loop AB 737:743 loop AB 737:743 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 1 19 theme Interferon 100:109 arg1 [also 120:124 Interferon (IFN)-λ1 [also 100:124 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] 100:154 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 3 20 theme IFN-λ1 402:407 arg1 structure 383:391 the structure 379:391 the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs 379:472 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 0 21 theme human 21:25 arg1 interferon-λ1 27:39 human interferon-λ1 21:39 human interferon-λ1 21:39 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 7 22 theme long-range 985:994 arg1 interactions 1002:1013 long-range ionic interactions 985:1013 long-range ionic interactions between ligand and receptor 985:1041 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 4 23 theme IL-10 583:587 arg1 family 589:594 the IL-10 family 579:594 the IL-10 family of cytokines 579:607 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 7 24 theme molecules 1092:1100 arg1 recognition 1073:1083 initial recognition 1065:1083 initial recognition of the molecules 1065:1100 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 5 25 theme fibronectin 658:668 arg1 topology 679:686 fibronectin type III topology 658:686 fibronectin type III topology 658:686 IFN-λR1 consists of two distinct domains having fibronectin type III topology. 20934432 3 26 theme type 460:463 arg1 IFNs 469:472 type III IFNs 460:472 type III IFNs 460:472 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 7 27 theme recognition 1073:1083 arg1 process 1054:1060 the process 1050:1060 the process of initial recognition of the molecules 1050:1100 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 2 28 theme receptor 306:313 arg1 complexes 315:323 specific heterodimeric receptor complexes 283:323 specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2 283:357 All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2. 20934432 5 29 theme type 670:673 arg1 topology 679:686 fibronectin type III topology 658:686 fibronectin type III topology 658:686 IFN-λR1 consists of two distinct domains having fibronectin type III topology. 20934432 7 30 theme hydrophobic 1108:1118 arg1 interactions 1120:1131 hydrophobic interactions 1108:1131 hydrophobic interactions 1108:1131 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 3 31 theme human 424:428 arg1 IFN-λR1 430:436 human IFN-λR1 424:436 human IFN-λR1 424:436 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 3 31 theme human 424:428 arg1 receptor 441:448 a receptor 439:448 a receptor unique to type III IFNs 439:472 We have determined the structure of human IFN-λ1 complexed with human IFN-λR1, a receptor unique to type III IFNs. 20934432 0 32 theme interferon-λ1 27:39 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 0:98 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 6 33 from loops 786:790 arg1 site 769:772 the IFN site 761:772 the IFN site 761:772 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 1 34 theme discovered 180:189 arg1 group 191:195 the recently discovered group 167:195 the recently discovered group of type III IFNs 167:212 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 1 34 theme discovered 180:189 arg1 IFNs 209:212 type III IFNs 200:212 type III IFNs 200:212 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 4 35 theme overall 479:485 arg1 similar 524:530 similar 524:530 similar 524:530 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 4 35 theme overall 479:485 arg1 structure 487:495 The overall structure 475:495 The overall structure of IFN-λ1 475:505 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 7 36 theme hydrogen 949:956 arg1 bonds 958:962 only a few direct hydrogen bonds 931:962 only a few direct hydrogen bonds 931:962 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 2 37 theme complexes 315:323 arg1 formation 270:278 formation 270:278 formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2 270:357 All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2. 20934432 7 38 theme initial 1065:1071 arg1 recognition 1073:1083 initial recognition 1065:1083 initial recognition of the molecules 1065:1100 Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it. 20934432 4 39 theme IFN-λ1 500:505 arg1 similar 524:530 similar 524:530 similar 524:530 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 4 39 theme IFN-λ1 500:505 arg1 structure 487:495 The overall structure 475:495 The overall structure of IFN-λ1 475:505 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 1 40 theme -λ1 116:118 arg1 [also 120:124 Interferon (IFN)-λ1 [also 100:124 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] 100:154 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 5 41 theme distinct 634:641 arg1 domains 643:649 two distinct domains 630:649 two distinct domains having fibronectin type III topology 630:686 IFN-λR1 consists of two distinct domains having fibronectin type III topology. 20934432 2 42 theme type 219:222 arg1 IFNs 228:231 All type III IFNs 215:231 All type III IFNs 215:231 All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2. 20934432 2 43 theme heterodimeric 292:304 arg1 complexes 315:323 specific heterodimeric receptor complexes 283:323 specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2 283:357 All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2. 20934432 4 44 theme other 562:566 arg1 members 568:574 other members 562:574 other members of the IL-10 family of cytokines 562:607 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 6 45 theme IFN 765:767 arg1 site 769:772 the IFN site 761:772 the IFN site 761:772 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 46 theme ligand-receptor 693:707 arg1 interface 709:717 The ligand-receptor interface 689:717 The ligand-receptor interface 689:717 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 2 47 theme specific 283:290 arg1 complexes 315:323 specific heterodimeric receptor complexes 283:323 specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2 283:357 All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-λR1 and IL-10R2. 20934432 0 48 theme high-affinity 61:73 arg1 interferon-λR1 84:97 its high-affinity receptor interferon-λR1 57:97 its high-affinity receptor interferon-λR1 57:97 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 4 49 theme family 589:594 arg1 members 568:574 other members 562:574 other members of the IL-10 family of cytokines 562:607 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 4 49 theme family 589:594 arg1 structure 539:547 the structure 535:547 the structure of IL-10 535:556 The overall structure of IFN-λ1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. 20934432 0 50 with complex 44:50 arg1 interferon-λR1 84:97 its high-affinity receptor interferon-λR1 57:97 its high-affinity receptor interferon-λR1 57:97 Crystal structure of human interferon-λ1 in complex with its high-affinity receptor interferon-λR1. 20934432 1 51 theme type 200:203 arg1 IFNs 209:212 type III IFNs 200:212 type III IFNs 200:212 Interferon (IFN)-λ1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. 20934432 6 52 theme hinge 846:850 arg1 region 852:857 inter-domain hinge region 833:857 inter-domain hinge region 833:857 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 53 theme IFN-λR1 862:868 arg1 IFN-λR1 862:868 IFN-λR1 862:868 IFN-λR1 862:868 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 53 theme IFN-λR1 862:868 arg1 region 852:857 inter-domain hinge region 833:857 inter-domain hinge region 833:857 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 20934432 6 53 theme IFN-λR1 862:868 arg1 domain 822:827 the N-terminal domain 807:827 the N-terminal domain 807:827 The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-λR1. 21827946 0 0 theme antibodies 66:75 arg1 function 29:36 the function 25:36 the function of two anti-VEGF receptor 2 antibodies 25:75 The structural basis for the function of two anti-VEGF receptor 2 antibodies. 21827946 2 1 from 6.64 384:387 arg1 complex 276:282 complex 276:282 complex with domain 3 of VEGFR2 276:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 1 2 theme antiangiogenic 137:150 arg1 IMC-1121B 112:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 1 2 theme antiangiogenic 137:150 arg1 drug 152:155 a promising antiangiogenic drug 125:155 a promising antiangiogenic drug being tested for treatment of breast and gastric cancer 125:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 2 3 with complex 276:282 arg1 VEGFR2 301:306 VEGFR2 301:306 VEGFR2 301:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 3 with complex 276:282 arg1 domain 289:294 domain 3 289:296 domain 3 of VEGFR2 289:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 3 4 theme domain 602:607 arg1 interface 611:619 the domain 3:domain 4 interface 589:619 the domain 3:domain 4 interface 589:619 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 2 5 theme fragment 264:271 arg1 6.64 384:387 6.64 384:387 6.64 384:387 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 5 theme fragment 264:271 arg1 structure 237:245 the structure 233:245 the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2 233:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 5 theme fragment 264:271 arg1 structure 324:332 the structure 320:332 the structure of a different neutralizing anti-VEGFR2 antibody 320:381 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 3 6 theme VEGFR2 475:480 arg1 domain 482:487 VEGFR2 domain 3 475:489 VEGFR2 domain 3 475:489 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 2 7 theme Fab 260:262 arg1 fragment 264:271 the 1121B Fab fragment 250:271 the 1121B Fab fragment in complex with domain 3 of VEGFR2 250:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 8 theme antibody 374:381 arg1 6.64 384:387 6.64 384:387 6.64 384:387 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 8 theme antibody 374:381 arg1 structure 237:245 the structure 233:245 the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2 233:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 8 theme antibody 374:381 arg1 structure 324:332 the structure 320:332 the structure of a different neutralizing anti-VEGFR2 antibody 320:381 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 9 theme 1121B 254:258 arg1 fragment 264:271 the 1121B Fab fragment 250:271 the 1121B Fab fragment in complex with domain 3 of VEGFR2 250:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 10 theme different 339:347 arg1 antibody 374:381 a different neutralizing anti-VEGFR2 antibody 337:381 a different neutralizing anti-VEGFR2 antibody 337:381 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 0 11 theme structural 4:13 arg1 basis 15:19 The structural basis 0:19 The structural basis for the function of two anti-VEGF receptor 2 antibodies 0:75 The structural basis for the function of two anti-VEGF receptor 2 antibodies. 21827946 4 12 theme essential 656:664 arg1 residues 647:654 residues 647:654 residues essential for VEGF, 1121B, and 6.64 binding 647:698 Mutagenesis reveals that residues essential for VEGF, 1121B, and 6.64 binding are nonoverlapping among the three contact patches. 21827946 1 13 theme anti-VEGF 82:90 arg1 IMC-1121B 112:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 1 13 theme anti-VEGF 82:90 arg1 drug 152:155 a promising antiangiogenic drug 125:155 a promising antiangiogenic drug being tested for treatment of breast and gastric cancer 125:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 3 14 theme opposite 458:465 arg1 ends 467:470 opposite ends 458:470 opposite ends of VEGFR2 domain 3 458:489 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 1 15 theme receptor 92:99 arg1 IMC-1121B 112:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 1 15 theme receptor 92:99 arg1 drug 152:155 a promising antiangiogenic drug 125:155 a promising antiangiogenic drug being tested for treatment of breast and gastric cancer 125:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 1 16 theme breast 187:192 arg1 treatment 174:182 treatment 174:182 treatment of breast and gastric cancer 174:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 3 17 theme Fab 436:438 arg1 fragments 440:448 The two Fab fragments 428:448 The two Fab fragments 428:448 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 2 18 theme VEGFR2 301:306 arg1 VEGFR2 301:306 VEGFR2 301:306 VEGFR2 301:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 18 theme VEGFR2 301:306 arg1 domain 289:294 domain 3 289:296 domain 3 of VEGFR2 289:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 3 19 theme VEGF 514:517 arg1 binding 519:525 VEGF binding 514:525 VEGF binding 514:525 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 2 20 from structure 237:245 arg1 complex 276:282 complex 276:282 complex with domain 3 of VEGFR2 276:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 21 theme anti-VEGFR2 362:372 arg1 antibody 374:381 a different neutralizing anti-VEGFR2 antibody 337:381 a different neutralizing anti-VEGFR2 antibody 337:381 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 22 theme VEGFR2 411:416 arg1 domain 418:423 VEGFR2 domain 3 411:425 VEGFR2 domain 3 411:425 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 1 23 theme gastric 198:204 arg1 cancer 206:211 gastric cancer 198:211 gastric cancer 198:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 3 24 theme receptor 553:560 arg1 dimerization 562:573 receptor dimerization 553:573 receptor dimerization 553:573 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 4 25 theme contact 735:741 arg1 patches 743:749 the three contact patches 725:749 the three contact patches 725:749 Mutagenesis reveals that residues essential for VEGF, 1121B, and 6.64 binding are nonoverlapping among the three contact patches. 21827946 2 26 from structure 324:332 arg1 complex 276:282 complex 276:282 complex with domain 3 of VEGFR2 276:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 3 27 theme domain 593:598 arg1 interface 611:619 the domain 3:domain 4 interface 589:619 the domain 3:domain 4 interface 589:619 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 1 28 theme antibody 103:110 arg1 IMC-1121B 112:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 1 28 theme antibody 103:110 arg1 drug 152:155 a promising antiangiogenic drug 125:155 a promising antiangiogenic drug being tested for treatment of breast and gastric cancer 125:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 3 29 theme domain 482:487 arg1 ends 467:470 opposite ends 458:470 opposite ends of VEGFR2 domain 3 458:489 The two Fab fragments bind at opposite ends of VEGFR2 domain 3; 1121B directly blocks VEGF binding, whereas 6.64 may prevent receptor dimerization by perturbing the domain 3:domain 4 interface. 21827946 1 30 theme cancer 206:211 arg1 treatment 174:182 treatment 174:182 treatment of breast and gastric cancer 174:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 4 31 theme 6.64 687:690 arg1 binding 692:698 6.64 binding 687:698 6.64 binding 687:698 Mutagenesis reveals that residues essential for VEGF, 1121B, and 6.64 binding are nonoverlapping among the three contact patches. 21827946 2 32 from fragment 264:271 arg1 complex 276:282 complex 276:282 complex with domain 3 of VEGFR2 276:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 0 33 theme receptor 55:62 arg1 antibodies 66:75 two anti-VEGF receptor 2 antibodies 41:75 two anti-VEGF receptor 2 antibodies 41:75 The structural basis for the function of two anti-VEGF receptor 2 antibodies. 21827946 2 34 theme neutralizing 349:360 arg1 antibody 374:381 a different neutralizing anti-VEGFR2 antibody 337:381 a different neutralizing anti-VEGFR2 antibody 337:381 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 35 with complex 398:404 arg1 domain 418:423 VEGFR2 domain 3 411:425 VEGFR2 domain 3 411:425 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 0 36 theme anti-VEGF 45:53 arg1 antibodies 66:75 two anti-VEGF receptor 2 antibodies 41:75 two anti-VEGF receptor 2 antibodies 41:75 The structural basis for the function of two anti-VEGF receptor 2 antibodies. 21827946 1 37 theme promising 127:135 arg1 IMC-1121B 112:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B 78:120 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 1 37 theme promising 127:135 arg1 drug 152:155 a promising antiangiogenic drug 125:155 a promising antiangiogenic drug being tested for treatment of breast and gastric cancer 125:211 The anti-VEGF receptor 2 antibody IMC-1121B is a promising antiangiogenic drug being tested for treatment of breast and gastric cancer. 21827946 2 38 from complex 276:282 arg1 6.64 384:387 6.64 384:387 6.64 384:387 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 38 from complex 276:282 arg1 structure 237:245 the structure 233:245 the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2 233:306 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 21827946 2 38 from complex 276:282 arg1 structure 324:332 the structure 320:332 the structure of a different neutralizing anti-VEGFR2 antibody 320:381 We have determined the structure of the 1121B Fab fragment in complex with domain 3 of VEGFR2, as well as the structure of a different neutralizing anti-VEGFR2 antibody, 6.64, also in complex with VEGFR2 domain 3. 3342889 5 0 theme heavy 647:651 arg1 sequences 630:638 N-terminal sequences 619:638 N-terminal sequences of the heavy and light chains 619:668 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 6 1 theme known 789:793 arg1 sequences 795:803 known sequences 789:803 known sequences of cathepsins H and L from other species 789:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 4 2 theme propeptide 485:494 arg1 chain 446:450 The glycosylated mini chain 424:450 The glycosylated mini chain 424:450 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 2 theme propeptide 485:494 arg1 fragment 469:476 a proteolytic fragment 455:476 a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 455:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 2 theme propeptide 485:494 arg1 Mr 565:566 an Mr 562:566 an Mr of 23720 562:575 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 3 3 theme enzyme 416:421 arg1 form 404:407 the single-chain form 387:407 the single-chain form of the enzyme 387:421 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 6 4 dep cathepsins 808:817 arg1 H 819:819 H 819:819 H 819:819 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 6 4 dep cathepsins 808:817 arg1 cathepsins 808:817 cathepsins H and L 808:825 cathepsins H and L from other species 808:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 6 4 dep cathepsins 808:817 arg1 L 825:825 L 825:825 L 825:825 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 7 5 theme other 942:946 arg1 proteinases 957:967 other cysteine proteinases 942:967 other cysteine proteinases of the papain superfamily 942:993 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 4 6 theme cathepsin 499:507 arg1 propeptide 485:494 the propeptide 481:494 the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 481:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 7 7 theme cysteine 948:955 arg1 proteinases 957:967 other cysteine proteinases 942:967 other cysteine proteinases of the papain superfamily 942:993 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 7 8 theme superfamily 983:993 arg1 proteinases 957:967 other cysteine proteinases 942:967 other cysteine proteinases of the papain superfamily 942:993 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 7 8 theme superfamily 983:993 arg1 EC 925:926 EC 3.4.22.1 925:935 EC 3.4.22.1 925:935 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 7 8 theme superfamily 983:993 arg1 B 922:922 cathepsin B 912:922 cathepsin B (EC 3.4.22.1) 912:936 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 1 9 theme kidney 104:109 arg1 cathepsin 111:119 human kidney cathepsin 98:119 human kidney cathepsin H (EC 3.4.22.16) 98:136 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 9 theme kidney 104:109 arg1 EC 124:125 EC 3.4.22.16 124:135 EC 3.4.22.16 124:135 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 5 10 theme N-terminal 619:628 arg1 sequences 630:638 N-terminal sequences 619:638 N-terminal sequences of the heavy and light chains 619:668 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 5 11 theme fragments 713:721 arg1 sequences 683:691 the sequences 679:691 the sequences of cyanogen bromide fragments of the heavy chain 679:740 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 2 12 theme 230 220:222 arg1 residues 224:231 230 residues 220:231 230 residues 220:231 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 5 13 theme heavy 730:734 arg1 chain 736:740 the heavy chain 726:740 the heavy chain 726:740 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 3 14 theme mini 318:321 arg1 chain 323:327 the light, heavy and mini chain 297:327 chain 323:327 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 5 15 theme light 657:661 arg1 sequences 630:638 N-terminal sequences 619:638 N-terminal sequences of the heavy and light chains 619:668 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 3 16 theme heavy 308:312 arg1 chain 323:327 the light, heavy and mini chain 297:327 chain 323:327 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 5 17 theme chain 736:740 arg1 fragments 713:721 cyanogen bromide fragments 696:721 cyanogen bromide fragments of the heavy chain 696:740 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 0 18 theme acid 6:9 arg1 sequences 11:19 Amino acid sequences 0:19 Amino acid sequences of the human kidney 0:39 Amino acid sequences of the human kidney cathepsins H and L. 3342889 4 19 theme proteolytic 457:467 arg1 chain 446:450 The glycosylated mini chain 424:450 The glycosylated mini chain 424:450 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 19 theme proteolytic 457:467 arg1 fragment 469:476 a proteolytic fragment 455:476 a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 455:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 7 20 theme homology 900:907 arg1 degree 881:886 a high degree 874:886 a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily 874:993 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 4 21 theme acid 544:547 arg1 residues 549:556 217 amino acid residues 534:556 217 amino acid residues 534:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 0 22 theme Amino 0:4 arg1 sequences 11:19 Amino acid sequences 0:19 Amino acid sequences of the human kidney 0:39 Amino acid sequences of the human kidney cathepsins H and L. 3342889 2 23 contain contains 211:218 arg2 residues 224:231 230 residues 220:231 230 residues 220:231 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 2 23 contain contains 211:218 arg1 H 209:209 Cathepsin H 199:209 Cathepsin H 199:209 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 1 24 theme complete 65:72 arg1 sequences 85:93 The complete amino acid sequences 61:93 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) 61:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 25 theme human 142:146 arg1 L 165:165 human kidney cathepsin L 142:165 human kidney cathepsin L (EC 3.4.22.15) 142:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 25 theme human 142:146 arg1 3.4.22.15 171:179 EC 3.4.22.15 168:179 EC 3.4.22.15 168:179 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 6 26 theme other 832:836 arg1 species 838:844 other species 832:844 other species 832:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 5 27 theme amino 582:586 arg1 sequence 593:600 Its amino acid sequence 578:600 Its amino acid sequence 578:600 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 1 28 dep cathepsin 111:119 arg1 H 121:121 H 121:121 human kidney cathepsin H (EC 3.4.22.16) 98:136 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 7 29 dep Cathepsins 847:856 arg1 H 858:858 H 858:858 H 858:858 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 7 29 dep Cathepsins 847:856 arg1 Cathepsins 847:856 Cathepsins H and L 847:864 Cathepsins H and L 847:864 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 7 29 dep Cathepsins 847:856 arg1 L 864:864 L 864:864 L 864:864 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 1 30 theme kidney 148:153 arg1 L 165:165 human kidney cathepsin L 142:165 human kidney cathepsin L (EC 3.4.22.15) 142:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 30 theme kidney 148:153 arg1 3.4.22.15 171:179 EC 3.4.22.15 168:179 EC 3.4.22.15 168:179 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 3 31 theme light 301:305 arg1 chain 323:327 the light, heavy and mini chain 297:327 chain 323:327 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 5 32 theme acid 588:591 arg1 sequence 593:600 Its amino acid sequence 578:600 Its amino acid sequence 578:600 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 4 33 dep cathepsin 499:507 arg1 H. 509:510 H. 509:510 H. 509:510 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 1 34 theme acid 80:83 arg1 sequences 85:93 The complete amino acid sequences 61:93 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) 61:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 35 theme cathepsin 155:163 arg1 L 165:165 human kidney cathepsin L 142:165 human kidney cathepsin L (EC 3.4.22.15) 142:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 35 theme cathepsin 155:163 arg1 3.4.22.15 171:179 EC 3.4.22.15 168:179 EC 3.4.22.15 168:179 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 0 36 theme human 28:32 arg1 kidney 34:39 the human kidney 24:39 the human kidney 24:39 Amino acid sequences of the human kidney cathepsins H and L. 3342889 6 37 with comparison 773:782 arg1 sequences 795:803 known sequences 789:803 known sequences of cathepsins H and L from other species 789:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 6 38 from species 838:844 arg1 H 819:819 H 819:819 H 819:819 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 6 38 from species 838:844 arg1 cathepsins 808:817 cathepsins H and L 808:825 cathepsins H and L from other species 808:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 6 38 from species 838:844 arg1 L 825:825 L 825:825 L 825:825 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 6 38 from species 838:844 arg1 sequences 795:803 known sequences 789:803 known sequences of cathepsins H and L from other species 789:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 3 39 theme cyanogen 358:365 arg1 bromide 367:373 bromide 367:373 bromide 367:373 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 1 40 theme L 165:165 arg1 sequences 85:93 The complete amino acid sequences 61:93 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) 61:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 4 41 theme 23720 571:575 arg1 chain 446:450 The glycosylated mini chain 424:450 The glycosylated mini chain 424:450 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 41 theme 23720 571:575 arg1 fragment 469:476 a proteolytic fragment 455:476 a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 455:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 41 theme 23720 571:575 arg1 Mr 565:566 an Mr 562:566 an Mr of 23720 562:575 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 2 42 contain has 237:239 arg2 Mr 244:245 an Mr 241:245 an Mr of 25116 241:254 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 2 42 contain has 237:239 arg1 H 209:209 Cathepsin H 199:209 Cathepsin H 199:209 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 7 43 theme cathepsin 912:920 arg1 EC 925:926 EC 3.4.22.1 925:935 EC 3.4.22.1 925:935 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 7 43 theme cathepsin 912:920 arg1 B 922:922 cathepsin B 912:922 cathepsin B (EC 3.4.22.1) 912:936 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 4 44 contain has 530:532 arg2 residues 549:556 217 amino acid residues 534:556 217 amino acid residues 534:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 44 contain has 530:532 arg1 L 528:528 Human cathepsin L 512:528 Human cathepsin L 512:528 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 3 45 theme bromide 367:373 arg1 cleavage 375:382 cyanogen bromide cleavage 358:382 cyanogen bromide cleavage of the single-chain form of the enzyme 358:421 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 4 46 theme glycosylated 428:439 arg1 chain 446:450 The glycosylated mini chain 424:450 The glycosylated mini chain 424:450 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 46 theme glycosylated 428:439 arg1 fragment 469:476 a proteolytic fragment 455:476 a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 455:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 7 47 theme sequence 891:898 arg1 homology 900:907 sequence homology 891:907 sequence homology 891:907 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 4 48 theme mini 441:444 arg1 chain 446:450 The glycosylated mini chain 424:450 The glycosylated mini chain 424:450 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 48 theme mini 441:444 arg1 fragment 469:476 a proteolytic fragment 455:476 a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 455:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 7 49 theme high 876:879 arg1 degree 881:886 a high degree 874:886 a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily 874:993 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 3 50 theme form 404:407 arg1 cleavage 375:382 cyanogen bromide cleavage 358:382 cyanogen bromide cleavage of the single-chain form of the enzyme 358:421 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 0 51 theme kidney 34:39 arg1 sequences 11:19 Amino acid sequences 0:19 Amino acid sequences of the human kidney 0:39 Amino acid sequences of the human kidney cathepsins H and L. 3342889 6 52 theme cathepsins 808:817 arg1 sequences 795:803 known sequences 789:803 known sequences of cathepsins H and L from other species 789:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 5 53 dep heavy 647:651 arg1 chains 663:668 chains 663:668 chains 663:668 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 4 54 dep propeptide 485:494 arg1 has 530:532 has 530:532 has 217 amino acid residues 530:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 7 55 theme papain 976:981 arg1 superfamily 983:993 the papain superfamily 972:993 the papain superfamily 972:993 Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 3.4.22.1) and other cysteine proteinases of the papain superfamily. 3342889 1 56 theme human 98:102 arg1 cathepsin 111:119 human kidney cathepsin 98:119 human kidney cathepsin H (EC 3.4.22.16) 98:136 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 56 theme human 98:102 arg1 EC 124:125 EC 3.4.22.16 124:135 EC 3.4.22.16 124:135 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 57 theme amino 74:78 arg1 sequences 85:93 The complete amino acid sequences 61:93 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) 61:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 58 theme EC 168:169 arg1 L 165:165 human kidney cathepsin L 142:165 human kidney cathepsin L (EC 3.4.22.15) 142:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 1 58 theme EC 168:169 arg1 3.4.22.15 171:179 EC 3.4.22.15 168:179 EC 3.4.22.15 168:179 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 2 59 theme Cathepsin 199:207 arg1 H 209:209 Cathepsin H 199:209 Cathepsin H 199:209 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 4 60 gly glycosylated 428:439 arg1 chain 446:450 The glycosylated mini chain 424:450 The glycosylated mini chain 424:450 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 4 60 gly glycosylated 428:439 arg1 fragment 469:476 a proteolytic fragment 455:476 a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues 455:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 2 61 theme 25116 250:254 arg1 Mr 244:245 an Mr 241:245 an Mr of 25116 241:254 Cathepsin H contains 230 residues and has an Mr of 25116. 3342889 4 62 theme amino 538:542 arg1 residues 549:556 217 amino acid residues 534:556 217 amino acid residues 534:556 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 5 63 theme bromide 705:711 arg1 fragments 713:721 cyanogen bromide fragments 696:721 cyanogen bromide fragments of the heavy chain 696:740 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 1 64 theme cathepsin 111:119 arg1 sequences 85:93 The complete amino acid sequences 61:93 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) 61:180 The complete amino acid sequences of human kidney cathepsin H (EC 3.4.22.16) and human kidney cathepsin L (EC 3.4.22.15) were determined. 3342889 3 65 theme single-chain 391:402 arg1 form 404:407 the single-chain form 387:407 the single-chain form of the enzyme 387:421 The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. 3342889 5 66 theme cyanogen 696:703 arg1 fragments 713:721 cyanogen bromide fragments 696:721 cyanogen bromide fragments of the heavy chain 696:740 Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. 3342889 4 67 theme Human 512:516 arg1 L 528:528 Human cathepsin L 512:528 Human cathepsin L 512:528 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 3342889 6 68 from sequences 795:803 arg1 species 838:844 other species 832:844 other species 832:844 The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. 3342889 4 69 theme cathepsin 518:526 arg1 L 528:528 Human cathepsin L 512:528 Human cathepsin L 512:528 The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. 19285951 0 0 theme ATP 88:90 arg1 synthase 92:99 the cell surface type ATP synthase 66:99 the cell surface type ATP synthase 66:99 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 0 1 from interaction 14:24 arg1 T-cells 104:110 T-cells 104:110 T-cells 104:110 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 3 2 theme HRG-binding 636:646 arg1 proteins 648:655 the HRG-binding proteins 632:655 the HRG-binding proteins 632:655 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 2 3 theme concanavalin 319:330 arg1 A 332:332 concanavalin A 319:332 concanavalin A (Con A) 319:340 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 3 theme concanavalin 319:330 arg1 A 339:339 Con A 335:339 Con A 335:339 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 3 4 theme finger 757:762 arg1 method 773:778 a peptide mass finger printing method 742:778 a peptide mass finger printing method 742:778 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 0 5 theme type 83:86 arg1 synthase 92:99 the cell surface type ATP synthase 66:99 the cell surface type ATP synthase 66:99 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 3 6 gly glycopeptide 698:709 arg2 glycopeptide 698:709 HRG-derived glycopeptide affinity chromatography 686:733 HRG-derived glycopeptide affinity chromatography 686:733 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 2 7 theme cellular 536:543 arg1 event 545:549 this cellular event 531:549 this cellular event 531:549 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 1 8 theme plasma 152:157 arg1 protein 159:165 a plasma protein 150:165 a plasma protein implicated in the innate immune system 150:204 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 1 8 theme plasma 152:157 arg1 glycoprotein 128:139 Histidine-rich glycoprotein 113:139 Histidine-rich glycoprotein (HRG) 113:145 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 3 9 theme printing 764:771 arg1 method 773:778 a peptide mass finger printing method 742:778 a peptide mass finger printing method 742:778 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 2 10 theme cell 457:460 arg1 glycosaminoglycan 470:486 cell surface glycosaminoglycan or Fcgamma receptors 457:507 glycosaminoglycan 470:486 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 3 11 theme present 559:565 arg1 study 567:571 the present study 555:571 the present study 555:571 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 4 12 theme ATP 828:830 arg1 synthase 832:839 mitochondrial ATP synthase 814:839 mitochondrial ATP synthase 814:839 HRG specifically interacted with mitochondrial ATP synthase with a dissociation constant of 66 nM. 19285951 4 13 theme dissociation 848:859 arg1 constant 861:868 a dissociation constant 846:868 a dissociation constant of 66 nM 846:877 HRG specifically interacted with mitochondrial ATP synthase with a dissociation constant of 66 nM. 19285951 5 14 theme alpha- 896:901 arg1 presence 884:891 The presence 880:891 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell 880:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 5 15 theme synthase 928:935 arg1 beta-subunits 907:919 beta-subunits 907:919 beta-subunits 907:919 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 5 15 theme synthase 928:935 arg1 alpha- 896:901 alpha- 896:901 alpha- 896:901 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 3 16 theme peptide 744:750 arg1 method 773:778 a peptide mass finger printing method 742:778 a peptide mass finger printing method 742:778 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 3 17 theme proteins 648:655 arg1 one 625:627 one 625:627 one 625:627 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 3 17 theme proteins 648:655 arg1 proteins 648:655 the HRG-binding proteins 632:655 the HRG-binding proteins 632:655 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 4 18 theme mitochondrial 814:826 arg1 synthase 832:839 mitochondrial ATP synthase 814:839 mitochondrial ATP synthase 814:839 HRG specifically interacted with mitochondrial ATP synthase with a dissociation constant of 66 nM. 19285951 3 19 theme mass 752:755 arg1 method 773:778 a peptide mass finger printing method 742:778 a peptide mass finger printing method 742:778 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 7 20 theme HRG/Con 1416:1422 arg1 A 1424:1424 HRG/Con A 1416:1424 HRG/Con A 1416:1424 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 21 theme cell 1236:1239 arg1 surface 1241:1247 the cell surface 1232:1247 the cell surface ATP synthase 1232:1260 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 2 22 theme morphological 354:366 arg1 change 368:373 a morphological change 352:373 a morphological change 352:373 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 5 23 theme beta-subunits 907:919 arg1 presence 884:891 The presence 880:891 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell 880:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 5 24 theme FACS 1026:1029 arg1 analysis 1031:1038 FACS analysis 1026:1038 FACS analysis 1026:1038 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 2 25 theme human 395:399 arg1 MOLT-4 422:427 the human leukemic T-cell line MOLT-4 391:427 the human leukemic T-cell line MOLT-4 391:427 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 26 theme MOLT-4 422:427 arg1 adhesion 379:386 adhesion 379:386 adhesion of the human leukemic T-cell line MOLT-4 to culture dishes 379:445 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 26 theme MOLT-4 422:427 arg1 change 368:373 a morphological change 352:373 a morphological change 352:373 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 0 27 theme affinity 5:12 arg1 interaction 14:24 High affinity interaction 0:24 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 0:111 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 2 28 gly glycopeptide 272:283 arg2 glycopeptide 272:283 the Arg23-Lys66 glycopeptide 256:283 the Arg23-Lys66 glycopeptide 256:283 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 7 29 theme morphological 1344:1356 arg1 changes 1358:1364 the morphological changes 1340:1364 the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A 1340:1424 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 30 located observed 1366:1373 arg2 changes 1358:1364 the morphological changes 1340:1364 the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A 1340:1424 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 30 located observed 1366:1373 arg1 cells 1385:1389 MOLT-4 cells 1378:1389 MOLT-4 cells 1378:1389 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 0 31 theme High 0:3 arg1 interaction 14:24 High affinity interaction 0:24 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 0:111 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 2 32 attach derived 285:291 arg2 HRG 248:250 HRG 248:250 HRG 248:250 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 32 attach derived 285:291 arg1 HRG 298:300 HRG 298:300 HRG 298:300 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 7 33 theme MOLT-4 1304:1309 arg1 cells 1311:1315 MOLT-4 cells 1304:1315 MOLT-4 cells 1304:1315 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 2 34 theme Con 335:337 arg1 A 332:332 concanavalin A 319:332 concanavalin A (Con A) 319:340 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 34 theme Con 335:337 arg1 A 339:339 Con A 335:339 Con A 335:339 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 7 35 theme MOLT-4 1378:1383 arg1 cells 1385:1389 MOLT-4 cells 1378:1389 MOLT-4 cells 1378:1389 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 3 36 theme T-cells 675:681 arg1 surface 664:670 the surface 660:670 the surface of T-cells 660:681 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 1 37 gly glycoprotein 128:139 arg1 protein 159:165 a plasma protein 150:165 a plasma protein implicated in the innate immune system 150:204 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 1 37 gly glycoprotein 128:139 arg1 HRG 142:144 HRG 142:144 HRG 142:144 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 1 37 gly glycoprotein 128:139 arg1 glycoprotein 128:139 Histidine-rich glycoprotein 113:139 Histidine-rich glycoprotein (HRG) 113:145 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 6 38 theme cells 1095:1099 arg1 changes 1077:1083 The HRG/Con A-induced morphological changes 1041:1083 The HRG/Con A-induced morphological changes of MOLT-4 cells 1041:1099 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 19285951 1 39 theme innate 185:190 arg1 system 199:204 the innate immune system 181:204 the innate immune system 181:204 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 6 40 theme monoclonal 1134:1143 arg1 antibody 1145:1152 a monoclonal antibody 1132:1152 a monoclonal antibody against the beta-subunit of ATP synthase 1132:1193 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 19285951 5 41 theme immunofluorescent 995:1011 arg1 staining 1013:1020 immunofluorescent staining 995:1020 immunofluorescent staining 995:1020 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 2 42 theme Arg23-Lys66 260:270 arg1 glycopeptide 272:283 the Arg23-Lys66 glycopeptide 256:283 the Arg23-Lys66 glycopeptide 256:283 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 1 43 theme immune 192:197 arg1 system 199:204 the innate immune system 181:204 the innate immune system 181:204 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 2 44 theme line 417:420 arg1 MOLT-4 422:427 the human leukemic T-cell line MOLT-4 391:427 the human leukemic T-cell line MOLT-4 391:427 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 4 45 theme nM 876:877 arg1 constant 861:868 a dissociation constant 846:868 a dissociation constant of 66 nM 846:877 HRG specifically interacted with mitochondrial ATP synthase with a dissociation constant of 66 nM. 19285951 5 46 theme plasma 944:949 arg1 membrane 951:958 the plasma membrane 940:958 the plasma membrane of MOLT-4 cell 940:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 7 47 theme binding 1277:1283 arg1 protein 1285:1291 a binding protein 1275:1291 a binding protein for HRG 1275:1299 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 47 theme binding 1277:1283 arg1 synthase 1253:1260 the cell surface ATP synthase 1232:1260 the cell surface ATP synthase 1232:1260 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 48 theme ATP 1249:1251 arg1 protein 1285:1291 a binding protein 1275:1291 a binding protein for HRG 1275:1299 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 48 theme ATP 1249:1251 arg1 synthase 1253:1260 the cell surface ATP synthase 1232:1260 the cell surface ATP synthase 1232:1260 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 0 49 theme histidine-rich 34:47 arg1 glycoprotein 49:60 histidine-rich glycoprotein 34:60 histidine-rich glycoprotein 34:60 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 3 50 theme glycopeptide 698:709 arg1 chromatography 720:733 HRG-derived glycopeptide affinity chromatography 686:733 HRG-derived glycopeptide affinity chromatography 686:733 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 2 51 theme T-cell 410:415 arg1 MOLT-4 422:427 the human leukemic T-cell line MOLT-4 391:427 the human leukemic T-cell line MOLT-4 391:427 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 52 with concert 306:312 arg1 A 332:332 concanavalin A 319:332 concanavalin A (Con A) 319:340 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 52 with concert 306:312 arg1 A 339:339 Con A 335:339 Con A 335:339 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 6 53 theme synthase 1186:1193 arg1 beta-subunit 1166:1177 the beta-subunit 1162:1177 the beta-subunit of ATP synthase 1162:1193 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 19285951 5 54 attach presence 884:891 arg1 membrane 951:958 the plasma membrane 940:958 the plasma membrane of MOLT-4 cell 940:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 5 54 attach presence 884:891 arg2 beta-subunits 907:919 beta-subunits 907:919 beta-subunits 907:919 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 5 54 attach presence 884:891 arg2 alpha- 896:901 alpha- 896:901 alpha- 896:901 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 2 55 theme leukemic 401:408 arg1 MOLT-4 422:427 the human leukemic T-cell line MOLT-4 391:427 the human leukemic T-cell line MOLT-4 391:427 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 3 56 theme affinity 711:718 arg1 chromatography 720:733 HRG-derived glycopeptide affinity chromatography 686:733 HRG-derived glycopeptide affinity chromatography 686:733 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 0 57 theme cell 70:73 arg1 synthase 92:99 the cell surface type ATP synthase 66:99 the cell surface type ATP synthase 66:99 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 2 58 theme recent 210:215 arg1 studies 217:223 recent studies 210:223 recent studies 210:223 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 3 59 link HRG-derived 686:696 arg1 chromatography 720:733 HRG-derived glycopeptide affinity chromatography 686:733 HRG-derived glycopeptide affinity chromatography 686:733 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 1 60 theme Histidine-rich 113:126 arg1 protein 159:165 a plasma protein 150:165 a plasma protein implicated in the innate immune system 150:204 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 1 60 theme Histidine-rich 113:126 arg1 HRG 142:144 HRG 142:144 HRG 142:144 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 1 60 theme Histidine-rich 113:126 arg1 glycoprotein 128:139 Histidine-rich glycoprotein 113:139 Histidine-rich glycoprotein (HRG) 113:145 Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. 19285951 2 61 theme surface 462:468 arg1 glycosaminoglycan 470:486 cell surface glycosaminoglycan or Fcgamma receptors 457:507 glycosaminoglycan 470:486 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 2 62 theme Fcgamma 491:497 arg1 receptors 499:507 cell surface glycosaminoglycan or Fcgamma receptors 457:507 receptors 499:507 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 5 63 theme cell 970:973 arg1 membrane 951:958 the plasma membrane 940:958 the plasma membrane of MOLT-4 cell 940:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 5 64 theme MOLT-4 963:968 arg1 cell 970:973 MOLT-4 cell 963:973 MOLT-4 cell 963:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 7 65 theme surface 1241:1247 arg1 protein 1285:1291 a binding protein 1275:1291 a binding protein for HRG 1275:1299 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 7 65 theme surface 1241:1247 arg1 synthase 1253:1260 the cell surface ATP synthase 1232:1260 the cell surface ATP synthase 1232:1260 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 5 66 from presence 884:891 arg1 membrane 951:958 the plasma membrane 940:958 the plasma membrane of MOLT-4 cell 940:973 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 3 67 theme HRG-derived 686:696 arg1 chromatography 720:733 HRG-derived glycopeptide affinity chromatography 686:733 HRG-derived glycopeptide affinity chromatography 686:733 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 3 68 theme synthase 613:620 arg1 alpha-subunit 592:604 the alpha-subunit 588:604 the alpha-subunit of ATP synthase 588:620 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 3 69 theme ATP 609:611 arg1 synthase 613:620 ATP synthase 609:620 ATP synthase 609:620 In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. 19285951 5 70 theme ATP 924:926 arg1 synthase 928:935 ATP synthase 924:935 ATP synthase 924:935 The presence of alpha- and beta-subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. 19285951 6 71 theme morphological 1063:1075 arg1 changes 1077:1083 The HRG/Con A-induced morphological changes 1041:1083 The HRG/Con A-induced morphological changes of MOLT-4 cells 1041:1099 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 19285951 7 72 with treatment 1401:1409 arg1 A 1424:1424 HRG/Con A 1416:1424 HRG/Con A 1416:1424 These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT-4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. 19285951 6 73 theme ATP 1182:1184 arg1 synthase 1186:1193 ATP synthase 1182:1193 ATP synthase 1182:1193 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 19285951 0 74 gly glycoprotein 49:60 arg1 glycoprotein 49:60 histidine-rich glycoprotein 34:60 histidine-rich glycoprotein 34:60 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 6 75 theme A-induced 1053:1061 arg1 changes 1077:1083 The HRG/Con A-induced morphological changes 1041:1083 The HRG/Con A-induced morphological changes of MOLT-4 cells 1041:1099 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 19285951 0 76 theme surface 75:81 arg1 synthase 92:99 the cell surface type ATP synthase 66:99 the cell surface type ATP synthase 66:99 High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. 19285951 2 77 theme culture 432:438 arg1 dishes 440:445 culture dishes 432:445 culture dishes 432:445 In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. 19285951 6 78 theme MOLT-4 1088:1093 arg1 cells 1095:1099 MOLT-4 cells 1088:1099 MOLT-4 cells 1088:1099 The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. 31310764 8 0 theme critical 1614:1621 arg1 assemblies 1603:1612 macromolecular assemblies 1588:1612 macromolecular assemblies critical to mucosal epithelia and the vasculature 1588:1662 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 4 1 theme module 716:721 arg1 structure 675:683 the x-ray crystal structure 657:683 the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds 657:787 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 8 2 theme module 1446:1451 arg1 structure 1453:1461 The mucin 2 module structure 1434:1461 The mucin 2 module structure 1434:1461 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 1 3 theme intestinal 164:173 arg1 epithelia 175:183 intestinal epithelia 164:183 intestinal epithelia 164:183 The mucin 2 glycoprotein assembles into a complex hydrogel that protects intestinal epithelia and houses the gut microbiome. 31310764 8 4 theme Willebrand 1516:1525 arg1 factor 1527:1532 von Willebrand factor 1512:1532 von Willebrand factor 1512:1532 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 7 5 theme residues 1168:1175 arg1 knowledge 1141:1149 knowledge 1141:1149 knowledge of the interface residues 1141:1175 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 6 from mucins 1258:1263 arg1 present 1241:1247 present 1241:1247 present 1241:1247 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 2 7 theme hydrogel 356:363 arg1 expansion 365:373 hydrogel expansion 356:373 hydrogel expansion 356:373 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 8 8 theme von 1512:1514 arg1 factor 1527:1532 von Willebrand factor 1512:1532 von Willebrand factor 1512:1532 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 7 9 theme interface 1158:1166 arg1 residues 1168:1175 the interface residues 1154:1175 the interface residues 1154:1175 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 10 theme acids 1226:1230 arg1 acids 1226:1230 these amino acids 1214:1230 these amino acids 1214:1230 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 10 theme acids 1226:1230 arg1 many 1206:1209 many 1206:1209 many 1206:1209 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 11 from factor 1287:1292 arg1 present 1241:1247 present 1241:1247 present 1241:1247 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 3 12 dep are 401:403 arg1 become 433:438 become 433:438 become covalently linked to one another 433:471 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 12 dep are 401:403 arg1 compares 505:512 compares 505:512 compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor 505:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 8 13 theme mucin 1438:1442 arg1 structure 1453:1461 The mucin 2 module structure 1434:1461 The mucin 2 module structure 1434:1461 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 2 14 theme dimers 295:300 arg1 step 224:227 A major step 216:227 A major step in mucin 2 assembly 216:247 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 2 14 theme dimers 295:300 arg1 multimerization 260:274 further multimerization 252:274 further multimerization 252:274 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 7 15 theme protein 1418:1424 arg1 family 1426:1431 this functionally broad protein family 1394:1431 this functionally broad protein family 1394:1431 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 3 16 theme Important 376:384 arg1 questions 391:399 Important open questions 376:399 Important open questions 376:399 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 17 from processes 529:537 arg1 factor 634:639 the hemostasis protein von Willebrand factor 596:639 the hemostasis protein von Willebrand factor 596:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 17 from processes 529:537 arg1 mucins 585:590 respiratory tract mucins 567:590 respiratory tract mucins 567:590 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 17 from processes 529:537 arg1 polymers 550:557 related polymers 542:557 related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor 542:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 2 18 theme mucin 289:293 arg1 dimers 295:300 preformed mucin dimers 279:300 preformed mucin dimers 279:300 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 4 19 theme multimerization 700:714 arg1 module 716:721 the mucin 2 multimerization module 688:721 the mucin 2 multimerization module 688:721 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 7 20 theme stable 1323:1328 arg1 arrangement 1336:1346 the stable dimer arrangement 1319:1346 the stable dimer arrangement reported herein 1319:1362 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 20 theme stable 1323:1328 arg1 likely 1367:1372 likely 1367:1372 likely 1367:1372 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 2 21 theme preformed 279:287 arg1 dimers 295:300 preformed mucin dimers 279:300 preformed mucin dimers 279:300 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 2 22 from step 224:227 arg1 assembly 240:247 mucin 2 assembly 232:247 mucin 2 assembly 232:247 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 5 23 theme mucin 863:867 arg1 assembly 869:876 intestinal mucin assembly 852:876 intestinal mucin assembly 852:876 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 1 24 gly glycoprotein 103:114 arg1 glycoprotein 103:114 The mucin 2 glycoprotein 91:114 The mucin 2 glycoprotein 91:114 The mucin 2 glycoprotein assembles into a complex hydrogel that protects intestinal epithelia and houses the gut microbiome. 31310764 4 25 theme disulfide 773:781 arg1 bonds 783:787 two intersubunit disulfide bonds 756:787 two intersubunit disulfide bonds 756:787 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 6 26 theme structure 1125:1133 arg1 relevance 1088:1096 the physiological relevance 1070:1096 the physiological relevance of the observed quaternary structure 1070:1133 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 4 27 theme crystal 667:673 arg1 structure 675:683 the x-ray crystal structure 657:683 the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds 657:787 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 0 28 theme Gel-Forming 11:21 arg1 Mucins 23:28 Intestinal Gel-Forming Mucins 0:28 Intestinal Gel-Forming Mucins 0:28 Intestinal Gel-Forming Mucins Polymerize by Disulfide-Mediated Dimerization of D3 Domains. 31310764 7 29 theme von 1272:1274 arg1 factor 1287:1292 von Willebrand factor 1272:1292 von Willebrand factor 1272:1292 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 1 30 theme mucin 95:99 arg1 glycoprotein 103:114 The mucin 2 glycoprotein 91:114 The mucin 2 glycoprotein 91:114 The mucin 2 glycoprotein assembles into a complex hydrogel that protects intestinal epithelia and houses the gut microbiome. 31310764 7 31 theme dimer 1330:1334 arg1 arrangement 1336:1346 the stable dimer arrangement 1319:1346 the stable dimer arrangement reported herein 1319:1362 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 31 theme dimer 1330:1334 arg1 likely 1367:1372 likely 1367:1372 likely 1367:1372 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 0 32 theme Intestinal 0:9 arg1 Mucins 23:28 Intestinal Gel-Forming Mucins 0:28 Intestinal Gel-Forming Mucins 0:28 Intestinal Gel-Forming Mucins Polymerize by Disulfide-Mediated Dimerization of D3 Domains. 31310764 8 33 theme structural 1559:1568 arg1 parallels 1570:1578 deep structural parallels 1554:1578 deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature 1554:1662 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 7 34 from present 1241:1247 arg1 factor 1287:1292 von Willebrand factor 1272:1292 von Willebrand factor 1272:1292 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 34 from present 1241:1247 arg1 mucins 1258:1263 other mucins 1252:1263 other mucins 1252:1263 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 4 35 theme intersubunit 760:771 arg1 bonds 783:787 two intersubunit disulfide bonds 756:787 two intersubunit disulfide bonds 756:787 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 1 36 theme gut 200:202 arg1 microbiome 204:213 the gut microbiome 196:213 the gut microbiome 196:213 The mucin 2 glycoprotein assembles into a complex hydrogel that protects intestinal epithelia and houses the gut microbiome. 31310764 3 37 theme open 386:389 arg1 questions 391:399 Important open questions 376:399 Important open questions 376:399 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 6 38 theme dimer 991:995 arg1 interface 997:1005 the dimer interface 987:1005 the dimer interface 987:1005 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 3 39 theme respiratory 567:577 arg1 mucins 585:590 respiratory tract mucins 567:590 respiratory tract mucins 567:590 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 5 40 theme current 834:840 arg1 model 842:846 the current model 830:846 the current model 830:846 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 3 41 theme related 542:548 arg1 factor 634:639 the hemostasis protein von Willebrand factor 596:639 the hemostasis protein von Willebrand factor 596:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 41 theme related 542:548 arg1 mucins 585:590 respiratory tract mucins 567:590 respiratory tract mucins 567:590 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 41 theme related 542:548 arg1 polymers 550:557 related polymers 542:557 related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor 542:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 42 theme tract 579:583 arg1 mucins 585:590 respiratory tract mucins 567:590 respiratory tract mucins 567:590 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 6 43 theme physiological 1074:1086 arg1 relevance 1088:1096 the physiological relevance 1070:1096 the physiological relevance of the observed quaternary structure 1070:1133 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 0 44 theme Disulfide-Mediated 44:61 arg1 Dimerization 63:74 Disulfide-Mediated Dimerization 44:74 Disulfide-Mediated Dimerization of D3 Domains 44:88 Intestinal Gel-Forming Mucins Polymerize by Disulfide-Mediated Dimerization of D3 Domains. 31310764 5 45 theme same 934:937 arg1 module 939:944 the same module 930:944 the same module 930:944 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 7 46 theme amino 1220:1224 arg1 acids 1226:1230 these amino acids 1214:1230 these amino acids 1214:1230 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 3 47 theme protein 611:617 arg1 factor 634:639 the hemostasis protein von Willebrand factor 596:639 the hemostasis protein von Willebrand factor 596:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 4 48 theme x-ray 661:665 arg1 structure 675:683 the x-ray crystal structure 657:683 the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds 657:787 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 6 49 theme quaternary 1114:1123 arg1 structure 1125:1133 the observed quaternary structure 1101:1133 the observed quaternary structure 1101:1133 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 7 50 theme broad 1412:1416 arg1 family 1426:1431 this functionally broad protein family 1394:1431 this functionally broad protein family 1394:1431 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 2 51 theme mucin 232:236 arg1 assembly 240:247 mucin 2 assembly 232:247 mucin 2 assembly 232:247 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 5 52 theme intestinal 852:861 arg1 assembly 869:876 intestinal mucin assembly 852:876 intestinal mucin assembly 852:876 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 3 53 theme multiple 409:416 arg1 dimers 426:431 multiple mucin 2 dimers 409:431 multiple mucin 2 dimers 409:431 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 8 54 theme macromolecular 1588:1601 arg1 assemblies 1603:1612 macromolecular assemblies 1588:1612 macromolecular assemblies critical to mucosal epithelia and the vasculature 1588:1662 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 7 55 theme other 1252:1256 arg1 mucins 1258:1263 other mucins 1252:1263 other mucins 1252:1263 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 56 theme Willebrand 1276:1285 arg1 factor 1287:1292 von Willebrand factor 1272:1292 von Willebrand factor 1272:1292 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 3 57 theme mucin 418:422 arg1 dimers 426:431 multiple mucin 2 dimers 409:431 multiple mucin 2 dimers 409:431 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 58 theme mucin 481:485 arg1 multimerization 489:503 mucin 2 multimerization 481:503 mucin 2 multimerization 481:503 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 2 59 theme major 218:222 arg1 step 224:227 A major step 216:227 A major step in mucin 2 assembly 216:247 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 2 59 theme major 218:222 arg1 multimerization 260:274 further multimerization 252:274 further multimerization 252:274 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 3 60 theme hemostasis 600:609 arg1 factor 634:639 the hemostasis protein von Willebrand factor 596:639 the hemostasis protein von Willebrand factor 596:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 61 theme analogous 519:527 arg1 processes 529:537 analogous processes 519:537 analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor 519:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 5 62 theme module 939:944 arg1 trimerization 913:925 disulfide-mediated trimerization 894:925 disulfide-mediated trimerization of the same module 894:944 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 6 63 theme Key 947:949 arg1 residues 951:958 Key residues 947:958 Key residues making interactions across the dimer interface 947:1005 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 5 64 theme dimer 794:798 arg1 structure 800:808 The dimer structure 790:808 The dimer structure 790:808 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 3 65 dep one 461:463 arg1 another 465:471 another 465:471 another 465:471 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 3 66 theme von 619:621 arg1 factor 634:639 the hemostasis protein von Willebrand factor 596:639 the hemostasis protein von Willebrand factor 596:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 1 67 theme complex 133:139 arg1 hydrogel 141:148 a complex hydrogel 131:148 a complex hydrogel that protects intestinal epithelia and houses the gut microbiome 131:213 The mucin 2 glycoprotein assembles into a complex hydrogel that protects intestinal epithelia and houses the gut microbiome. 31310764 7 68 located present 1241:1247 arg1 mucins 1258:1263 other mucins 1252:1263 other mucins 1252:1263 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 68 located present 1241:1247 arg2 many 1206:1209 many 1206:1209 many 1206:1209 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 68 located present 1241:1247 arg2 acids 1226:1230 these amino acids 1214:1230 these amino acids 1214:1230 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 2 69 theme further 252:258 arg1 step 224:227 A major step 216:227 A major step in mucin 2 assembly 216:247 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 2 69 theme further 252:258 arg1 multimerization 260:274 further multimerization 252:274 further multimerization 252:274 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 5 70 theme disulfide-mediated 894:911 arg1 trimerization 913:925 disulfide-mediated trimerization 894:925 disulfide-mediated trimerization of the same module 894:944 The dimer structure calls into question the current model for intestinal mucin assembly, which proposes disulfide-mediated trimerization of the same module. 31310764 2 71 theme honeycomb-like 324:337 arg1 arrangement 339:349 a honeycomb-like arrangement 322:349 a honeycomb-like arrangement upon hydrogel expansion 322:373 A major step in mucin 2 assembly is further multimerization of preformed mucin dimers, thought to produce a honeycomb-like arrangement upon hydrogel expansion. 31310764 0 72 theme Domains 82:88 arg1 Dimerization 63:74 Disulfide-Mediated Dimerization 44:74 Disulfide-Mediated Dimerization of D3 Domains 44:88 Intestinal Gel-Forming Mucins Polymerize by Disulfide-Mediated Dimerization of D3 Domains. 31310764 3 73 theme Willebrand 623:632 arg1 factor 634:639 the hemostasis protein von Willebrand factor 596:639 the hemostasis protein von Willebrand factor 596:639 Important open questions are how multiple mucin 2 dimers become covalently linked to one another and how mucin 2 multimerization compares with analogous processes in related polymers such as respiratory tract mucins and the hemostasis protein von Willebrand factor. 31310764 8 74 theme deep 1554:1557 arg1 parallels 1570:1578 deep structural parallels 1554:1578 deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature 1554:1662 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 6 75 theme observed 1105:1112 arg1 structure 1125:1133 the observed quaternary structure 1101:1133 the observed quaternary structure 1101:1133 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 6 76 theme mucin 1042:1046 arg1 orthologs 1048:1056 intestinal mucin orthologs 1031:1056 intestinal mucin orthologs 1031:1056 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 31310764 0 77 theme D3 79:80 arg1 Domains 82:88 D3 Domains 79:88 D3 Domains 79:88 Intestinal Gel-Forming Mucins Polymerize by Disulfide-Mediated Dimerization of D3 Domains. 31310764 8 78 theme mucosal 1626:1632 arg1 epithelia 1634:1642 mucosal epithelia 1626:1642 mucosal epithelia 1626:1642 The mucin 2 module structure thus reveals the manner by which both mucins and von Willebrand factor polymerize, drawing deep structural parallels between macromolecular assemblies critical to mucosal epithelia and the vasculature. 31310764 7 79 attach present 1241:1247 arg1 factor 1287:1292 von Willebrand factor 1272:1292 von Willebrand factor 1272:1292 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 79 attach present 1241:1247 arg1 mucins 1258:1263 other mucins 1252:1263 other mucins 1252:1263 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 79 attach present 1241:1247 arg2 many 1206:1209 many 1206:1209 many 1206:1209 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 7 79 attach present 1241:1247 arg2 acids 1226:1230 these amino acids 1214:1230 these amino acids 1214:1230 With knowledge of the interface residues, it can be demonstrated that many of these amino acids are also present in other mucins and in von Willebrand factor, further indicating that the stable dimer arrangement reported herein is likely to be shared across this functionally broad protein family. 31310764 4 80 theme mucin 692:696 arg1 module 716:721 the mucin 2 multimerization module 688:721 the mucin 2 multimerization module 688:721 Here we report the x-ray crystal structure of the mucin 2 multimerization module, found to form a dimer linked by two intersubunit disulfide bonds. 31310764 6 81 theme intestinal 1031:1040 arg1 orthologs 1048:1056 intestinal mucin orthologs 1031:1056 intestinal mucin orthologs 1031:1056 Key residues making interactions across the dimer interface are highly conserved in intestinal mucin orthologs, supporting the physiological relevance of the observed quaternary structure. 11602603 2 0 theme protease 531:538 arg1 genes 540:544 serine protease genes 524:544 serine protease genes 524:544 The tryptase epsilon gene resides on chromosome 16p13.3 within a 2.5-Mb complex of serine protease genes. 11602603 7 1 theme tryptase 1118:1125 arg1 product 1146:1152 a major product 1138:1152 a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines 1138:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 1 theme tryptase 1118:1125 arg1 epsilon 1127:1133 tryptase epsilon 1118:1133 tryptase epsilon 1118:1133 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 11 2 from functions 1887:1895 arg1 humans 1900:1905 humans 1900:1905 humans 1900:1905 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 1 3 dep tryptase 373:380 arg1 epsilon 382:388 epsilon 382:388 epsilon 382:388 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 10 4 theme protein 1586:1592 arg1 modeling 1604:1611 comparative protein structure modeling 1574:1611 comparative protein structure modeling 1574:1611 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 4 5 theme epsilon 786:792 arg1 transcript 794:803 the tryptase epsilon transcript 773:803 the tryptase epsilon transcript 773:803 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 6 6 theme human 1073:1077 arg1 genes 1088:1092 other human tryptase genes 1067:1092 other human tryptase genes 1067:1092 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 1 7 theme human 217:221 arg1 cDNAs 232:236 varied human tryptase cDNAs 210:236 varied human tryptase cDNAs 210:236 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 8 theme S1 410:411 arg1 tryptase 373:380 tryptase epsilon or protease 373:400 tryptase 373:380 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 8 theme S1 410:411 arg1 member 420:425 serine S1 family member 22 403:428 serine S1 family member 22 (PRSS22) 403:437 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 8 theme S1 410:411 arg1 PRSS22 431:436 PRSS22 431:436 PRSS22 431:436 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 10 9 theme functional 1617:1626 arg1 studies 1628:1634 functional studies 1617:1634 functional studies using recombinant material 1617:1661 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 4 10 theme tryptase 777:784 arg1 transcript 794:803 the tryptase epsilon transcript 773:803 the tryptase epsilon transcript 773:803 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 3 11 theme tryptase 647:654 arg1 gene 669:672 tryptase epsilon-like gene 647:672 only one tryptase epsilon-like gene 638:672 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 0 12 theme human 82:86 arg1 proteases 95:103 human serine proteases 82:103 human serine proteases expressed in airway epithelial cells 82:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 1 13 theme novel 333:337 arg1 protease 352:359 a novel human serine protease 331:359 a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 331:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 0 14 theme proteases 95:103 arg1 family 72:77 the chromosome 16p13.3 family 49:77 the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells 49:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 1 15 theme serine 345:350 arg1 protease 352:359 a novel human serine protease 331:359 a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 331:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 11 16 theme genes 1853:1857 arg1 complex 1833:1839 the chromosome 16p13.3 complex 1810:1839 the chromosome 16p13.3 complex of tryptase genes 1810:1857 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 1 17 theme tag 185:187 arg1 base 200:203 sequence tag (EST) data base 176:203 sequence tag (EST) data base 176:203 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 2 18 theme 2.5-Mb 506:511 arg1 complex 513:519 a 2.5-Mb complex 504:519 a 2.5-Mb complex of serine protease genes 504:544 The tryptase epsilon gene resides on chromosome 16p13.3 within a 2.5-Mb complex of serine protease genes. 11602603 5 19 theme tryptase 835:842 arg1 transcript 852:861 the tryptase epsilon transcript 831:861 the tryptase epsilon transcript 831:861 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 7 20 theme cell 1233:1236 arg1 lines 1238:1242 varied transformed epithelial cell lines 1203:1242 varied transformed epithelial cell lines 1203:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 0 21 theme epithelial 125:134 arg1 cells 136:140 airway epithelial cells 118:140 airway epithelial cells 118:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 10 22 theme recombinant 1642:1652 arg1 material 1654:1661 recombinant material 1642:1661 recombinant material 1642:1661 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 7 23 theme transformed 1210:1220 arg1 lines 1238:1242 varied transformed epithelial cell lines 1203:1242 varied transformed epithelial cell lines 1203:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 24 theme cells 1185:1189 arg1 product 1146:1152 a major product 1138:1152 a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines 1138:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 24 theme cells 1185:1189 arg1 epsilon 1127:1133 tryptase epsilon 1118:1133 tryptase epsilon 1118:1133 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 0 25 theme new 35:37 arg1 epsilon 15:21 Human tryptase epsilon 0:21 Human tryptase epsilon (PRSS22) 0:30 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 0 25 theme new 35:37 arg1 member 39:44 a new member 33:44 a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells 33:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 7 26 theme cellular 1102:1109 arg1 level 1111:1115 the cellular level 1098:1115 the cellular level 1098:1115 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 2 27 theme epsilon 454:460 arg1 gene 462:465 The tryptase epsilon gene 441:465 The tryptase epsilon gene 441:465 The tryptase epsilon gene resides on chromosome 16p13.3 within a 2.5-Mb complex of serine protease genes. 11602603 0 28 theme chromosome 53:62 arg1 family 72:77 the chromosome 16p13.3 family 49:77 the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells 49:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 1 29 theme overlapping 307:317 arg1 portions 319:326 overlapping portions 307:326 overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 307:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 29 theme overlapping 307:317 arg1 ESTs 263:266 two truncated ESTs 249:266 two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 249:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 9 30 theme tryptase 1421:1428 arg1 alpha 1430:1434 human tryptase alpha 1415:1434 human tryptase alpha 1415:1434 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 6 31 from that 1059:1062 arg1 different 1044:1052 different 1044:1052 different 1044:1052 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 6 31 from that 1059:1062 arg1 manner 1029:1034 a developmentally regulated manner 1001:1034 a developmentally regulated manner that is different from that of other human tryptase genes 1001:1092 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 0 32 theme family 72:77 arg1 epsilon 15:21 Human tryptase epsilon 0:21 Human tryptase epsilon (PRSS22) 0:30 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 0 32 theme family 72:77 arg1 member 39:44 a new member 33:44 a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells 33:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 4 33 from levels 763:768 arg1 humans 814:819 adult humans 808:819 adult humans 808:819 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 7 34 theme varied 1203:1208 arg1 lines 1238:1242 varied transformed epithelial cell lines 1203:1242 varied transformed epithelial cell lines 1203:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 0 35 theme Human 0:4 arg1 epsilon 15:21 Human tryptase epsilon 0:21 Human tryptase epsilon (PRSS22) 0:30 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 0 35 theme Human 0:4 arg1 member 39:44 a new member 33:44 a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells 33:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 0 35 theme Human 0:4 arg1 PRSS22 24:29 PRSS22 24:29 PRSS22 24:29 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 1 36 theme tryptase 223:230 arg1 cDNAs 232:236 varied human tryptase cDNAs 210:236 varied human tryptase cDNAs 210:236 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 6 37 theme epsilon 957:963 arg1 gene 965:968 the tryptase epsilon gene 944:968 the tryptase epsilon gene 944:968 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 8 38 theme tryptase 1266:1273 arg1 epsilon 1275:1281 Enzymatically active tryptase epsilon 1245:1281 Enzymatically active tryptase epsilon 1245:1281 Enzymatically active tryptase epsilon is also constitutively secreted from these cells. 11602603 11 39 theme chromosome 1814:1823 arg1 complex 1833:1839 the chromosome 16p13.3 complex 1810:1839 the chromosome 16p13.3 complex of tryptase genes 1810:1857 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 5 40 theme adult 877:881 arg1 lung 889:892 adult human lung 877:892 adult human lung 877:892 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 10 41 from that 1735:1738 arg1 distinct 1721:1728 distinct 1721:1728 distinct 1721:1728 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 10 42 theme other 1747:1751 arg1 members 1760:1766 its other family members 1743:1766 its other family members 1743:1766 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 1 43 theme truncated 253:261 arg1 ESTs 263:266 two truncated ESTs 249:266 two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 249:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 43 theme truncated 253:261 arg1 portions 319:326 overlapping portions 307:326 overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 307:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 5 44 theme fetal 926:930 arg1 lung 932:935 fetal lung 926:935 fetal lung 926:935 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 7 45 theme epithelial 1174:1183 arg1 cells 1185:1189 normal pulmonary epithelial cells 1157:1189 normal pulmonary epithelial cells 1157:1189 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 4 46 theme highest 742:748 arg1 levels 763:768 the highest steady-state levels 738:768 the highest steady-state levels of the tryptase epsilon transcript in adult humans 738:819 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 10 47 theme comparative 1574:1584 arg1 modeling 1604:1611 comparative protein structure modeling 1574:1611 comparative protein structure modeling 1574:1611 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 6 48 theme regulated 1019:1027 arg1 different 1044:1052 different 1044:1052 different 1044:1052 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 6 48 theme regulated 1019:1027 arg1 manner 1029:1034 a developmentally regulated manner 1001:1034 a developmentally regulated manner that is different from that of other human tryptase genes 1001:1092 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 6 49 theme tryptase 1079:1086 arg1 genes 1088:1092 other human tryptase genes 1067:1092 other human tryptase genes 1067:1092 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 1 50 with Probing 143:149 arg1 cDNAs 232:236 varied human tryptase cDNAs 210:236 varied human tryptase cDNAs 210:236 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 51 dep protease 352:359 arg1 designated 362:371 designated 362:371 designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22) 362:437 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 3 52 theme genes 577:581 arg1 genes 577:581 the 14 genes 570:581 the 14 genes in this complex 570:597 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 3 52 theme genes 577:581 arg1 7 565:565 7 565:565 7 565:565 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 3 52 theme genes 577:581 arg1 proteases 627:635 enzymatically active proteases 606:635 enzymatically active proteases 606:635 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 7 53 theme normal 1157:1162 arg1 cells 1185:1189 normal pulmonary epithelial cells 1157:1189 normal pulmonary epithelial cells 1157:1189 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 3 54 theme active 620:625 arg1 genes 577:581 the 14 genes 570:581 the 14 genes in this complex 570:597 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 3 54 theme active 620:625 arg1 7 565:565 7 565:565 7 565:565 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 3 54 theme active 620:625 arg1 proteases 627:635 enzymatically active proteases 606:635 enzymatically active proteases 606:635 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 10 55 theme structure 1594:1602 arg1 modeling 1604:1611 comparative protein structure modeling 1574:1611 comparative protein structure modeling 1574:1611 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 5 56 located present 902:908 arg2 it 895:896 it 895:896 it 895:896 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 5 56 located present 902:908 arg1 abundance 913:921 abundance 913:921 abundance in fetal lung 913:935 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 2 57 theme serine 524:529 arg1 genes 540:544 serine protease genes 524:544 serine protease genes 524:544 The tryptase epsilon gene resides on chromosome 16p13.3 within a 2.5-Mb complex of serine protease genes. 11602603 9 58 theme transmembrane 1491:1503 arg1 gamma 1523:1527 transmembrane tryptase/tryptase gamma 1491:1527 transmembrane tryptase/tryptase gamma 1491:1527 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 1 59 theme varied 210:215 arg1 cDNAs 232:236 varied human tryptase cDNAs 210:236 varied human tryptase cDNAs 210:236 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 4 60 theme transcript 794:803 arg1 levels 763:768 the highest steady-state levels 738:768 the highest steady-state levels of the tryptase epsilon transcript in adult humans 738:819 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 6 61 theme other 1067:1071 arg1 genes 1088:1092 other human tryptase genes 1067:1092 other human tryptase genes 1067:1092 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 1 62 theme serine 403:408 arg1 tryptase 373:380 tryptase epsilon or protease 373:400 tryptase 373:380 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 62 theme serine 403:408 arg1 member 420:425 serine S1 family member 22 403:428 serine S1 family member 22 (PRSS22) 403:437 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 62 theme serine 403:408 arg1 PRSS22 431:436 PRSS22 431:436 PRSS22 431:436 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 11 63 theme varied 1880:1885 arg1 functions 1887:1895 varied functions 1880:1895 varied functions in humans 1880:1905 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 5 64 from abundance 913:921 arg1 lung 932:935 fetal lung 926:935 fetal lung 926:935 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 5 64 from abundance 913:921 arg1 present 902:908 present 902:908 present 902:908 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 4 65 theme steady-state 750:761 arg1 levels 763:768 the highest steady-state levels 738:768 the highest steady-state levels of the tryptase epsilon transcript in adult humans 738:819 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 1 66 theme family 413:418 arg1 tryptase 373:380 tryptase epsilon or protease 373:400 tryptase 373:380 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 66 theme family 413:418 arg1 member 420:425 serine S1 family member 22 403:428 serine S1 family member 22 (PRSS22) 403:437 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 66 theme family 413:418 arg1 PRSS22 431:436 PRSS22 431:436 PRSS22 431:436 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 10 67 theme tryptase 1677:1684 arg1 epsilon 1686:1692 tryptase epsilon 1677:1692 tryptase epsilon 1677:1692 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 5 68 from present 902:908 arg1 abundance 913:921 abundance 913:921 abundance in fetal lung 913:935 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 9 69 theme amino 1337:1341 arg1 identical 1393:1401 identical 1393:1401 identical 1393:1401 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 9 69 theme amino 1337:1341 arg1 sequence 1348:1355 The amino acid sequence 1333:1355 The amino acid sequence of human tryptase epsilon 1333:1381 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 2 70 theme genes 540:544 arg1 complex 513:519 a 2.5-Mb complex 504:519 a 2.5-Mb complex of serine protease genes 504:544 The tryptase epsilon gene resides on chromosome 16p13.3 within a 2.5-Mb complex of serine protease genes. 11602603 0 71 theme serine 88:93 arg1 proteases 95:103 human serine proteases 82:103 human serine proteases expressed in airway epithelial cells 82:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 1 72 theme GenBank 158:164 arg1 Probing 143:149 Probing 143:149 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs 143:236 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 11 73 contain carry 1870:1874 arg1 products 1798:1805 the products 1794:1805 the products of the chromosome 16p13.3 complex of tryptase genes 1794:1857 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 11 73 contain carry 1870:1874 arg2 functions 1887:1895 varied functions 1880:1895 varied functions in humans 1880:1905 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 3 74 theme epsilon-like 656:667 arg1 gene 669:672 tryptase epsilon-like gene 647:672 only one tryptase epsilon-like gene 638:672 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 1 75 theme human 339:343 arg1 protease 352:359 a novel human serine protease 331:359 a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 331:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 10 76 theme substrate 1700:1708 arg1 preference 1710:1719 a substrate preference 1698:1719 a substrate preference distinct from that of its other family members 1698:1766 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 9 77 theme acid 1343:1346 arg1 identical 1393:1401 identical 1393:1401 identical 1393:1401 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 9 77 theme acid 1343:1346 arg1 sequence 1348:1355 The amino acid sequence 1333:1355 The amino acid sequence of human tryptase epsilon 1333:1381 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 1 78 theme sequence 176:183 arg1 EST 190:192 EST 190:192 EST 190:192 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 78 theme sequence 176:183 arg1 tag 185:187 sequence tag 176:187 sequence tag (EST) data base 176:203 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 0 79 theme airway 118:123 arg1 cells 136:140 airway epithelial cells 118:140 airway epithelial cells 118:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 1 80 theme protease 352:359 arg1 portions 319:326 overlapping portions 307:326 overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 307:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 1 80 theme protease 352:359 arg1 ESTs 263:266 two truncated ESTs 249:266 two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)) 249:438 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 11602603 11 81 theme complex 1833:1839 arg1 products 1798:1805 the products 1794:1805 the products of the chromosome 16p13.3 complex of tryptase genes 1794:1857 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 9 82 theme epsilon 1375:1381 arg1 identical 1393:1401 identical 1393:1401 identical 1393:1401 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 9 82 theme epsilon 1375:1381 arg1 sequence 1348:1355 The amino acid sequence 1333:1355 The amino acid sequence of human tryptase epsilon 1333:1381 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 4 83 theme adult 808:812 arg1 humans 814:819 adult humans 808:819 adult humans 808:819 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 5 84 theme epsilon 844:850 arg1 transcript 852:861 the tryptase epsilon transcript 831:861 the tryptase epsilon transcript 831:861 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 7 85 from level 1111:1115 arg1 product 1146:1152 a major product 1138:1152 a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines 1138:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 85 from level 1111:1115 arg1 epsilon 1127:1133 tryptase epsilon 1118:1133 tryptase epsilon 1118:1133 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 86 theme lines 1238:1242 arg1 product 1146:1152 a major product 1138:1152 a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines 1138:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 86 theme lines 1238:1242 arg1 epsilon 1127:1133 tryptase epsilon 1118:1133 tryptase epsilon 1118:1133 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 10 87 theme distinct 1721:1728 arg1 preference 1710:1719 a substrate preference 1698:1719 a substrate preference distinct from that of its other family members 1698:1766 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 7 88 theme epithelial 1222:1231 arg1 lines 1238:1242 varied transformed epithelial cell lines 1203:1242 varied transformed epithelial cell lines 1203:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 5 89 attach present 902:908 arg2 it 895:896 it 895:896 it 895:896 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 5 89 attach present 902:908 arg1 abundance 913:921 abundance 913:921 abundance in fetal lung 913:935 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 5 90 theme human 883:887 arg1 lung 889:892 adult human lung 877:892 adult human lung 877:892 Although the tryptase epsilon transcript was scarce in adult human lung, it was present in abundance in fetal lung. 11602603 2 91 theme tryptase 445:452 arg1 gene 462:465 The tryptase epsilon gene 441:465 The tryptase epsilon gene 441:465 The tryptase epsilon gene resides on chromosome 16p13.3 within a 2.5-Mb complex of serine protease genes. 11602603 4 92 contain contain 730:736 arg1 esophagus 706:714 esophagus 706:714 esophagus 706:714 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 4 92 contain contain 730:736 arg2 levels 763:768 the highest steady-state levels 738:768 the highest steady-state levels of the tryptase epsilon transcript in adult humans 738:819 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 4 92 contain contain 730:736 arg1 trachea 694:700 trachea 694:700 trachea 694:700 The trachea and esophagus were found to contain the highest steady-state levels of the tryptase epsilon transcript in adult humans. 11602603 0 93 theme tryptase 6:13 arg1 epsilon 15:21 Human tryptase epsilon 0:21 Human tryptase epsilon (PRSS22) 0:30 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 0 93 theme tryptase 6:13 arg1 member 39:44 a new member 33:44 a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells 33:140 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 0 93 theme tryptase 6:13 arg1 PRSS22 24:29 PRSS22 24:29 PRSS22 24:29 Human tryptase epsilon (PRSS22), a new member of the chromosome 16p13.3 family of human serine proteases expressed in airway epithelial cells. 11602603 9 94 theme tryptase/tryptase 1505:1521 arg1 gamma 1523:1527 transmembrane tryptase/tryptase gamma 1491:1527 transmembrane tryptase/tryptase gamma 1491:1527 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 9 95 theme tryptase 1437:1444 arg1 beta 1446:1449 tryptase beta I 1437:1451 tryptase beta I 1437:1451 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 10 96 contain has 1694:1696 arg1 epsilon 1686:1692 tryptase epsilon 1677:1692 tryptase epsilon 1677:1692 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 10 96 contain has 1694:1696 arg2 preference 1710:1719 a substrate preference 1698:1719 a substrate preference distinct from that of its other family members 1698:1766 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 11 97 theme tryptase 1844:1851 arg1 genes 1853:1857 tryptase genes 1844:1857 tryptase genes 1844:1857 These data indicate that the products of the chromosome 16p13.3 complex of tryptase genes evolved to carry out varied functions in humans. 11602603 6 98 theme tryptase 948:955 arg1 gene 965:968 the tryptase epsilon gene 944:968 the tryptase epsilon gene 944:968 Thus, the tryptase epsilon gene is expressed in the airways in a developmentally regulated manner that is different from that of other human tryptase genes. 11602603 10 99 theme family 1753:1758 arg1 members 1760:1766 its other family members 1743:1766 its other family members 1743:1766 Nevertheless, comparative protein structure modeling and functional studies using recombinant material revealed that tryptase epsilon has a substrate preference distinct from that of its other family members. 11602603 8 100 theme active 1259:1264 arg1 epsilon 1275:1281 Enzymatically active tryptase epsilon 1245:1281 Enzymatically active tryptase epsilon 1245:1281 Enzymatically active tryptase epsilon is also constitutively secreted from these cells. 11602603 9 101 theme human 1360:1364 arg1 epsilon 1375:1381 human tryptase epsilon 1360:1381 human tryptase epsilon 1360:1381 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 9 102 theme tryptase 1454:1461 arg1 beta 1463:1466 tryptase beta II 1454:1469 tryptase beta II 1454:1469 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 7 103 theme pulmonary 1164:1172 arg1 cells 1185:1189 normal pulmonary epithelial cells 1157:1189 normal pulmonary epithelial cells 1157:1189 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 9 104 theme human 1415:1419 arg1 alpha 1430:1434 human tryptase alpha 1415:1434 human tryptase alpha 1415:1434 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 9 105 theme tryptase 1472:1479 arg1 beta 1481:1484 tryptase beta III 1472:1488 tryptase beta III 1472:1488 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 7 106 theme major 1140:1144 arg1 product 1146:1152 a major product 1138:1152 a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines 1138:1242 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 7 106 theme major 1140:1144 arg1 epsilon 1127:1133 tryptase epsilon 1118:1133 tryptase epsilon 1118:1133 At the cellular level, tryptase epsilon is a major product of normal pulmonary epithelial cells, as well as varied transformed epithelial cell lines. 11602603 9 107 theme tryptase 1366:1373 arg1 epsilon 1375:1381 human tryptase epsilon 1360:1381 human tryptase epsilon 1360:1381 The amino acid sequence of human tryptase epsilon is 38-44% identical to those of human tryptase alpha, tryptase beta I, tryptase beta II, tryptase beta III, transmembrane tryptase/tryptase gamma, marapsin, and Esp-1/testisin. 11602603 3 108 from genes 577:581 arg1 complex 591:597 this complex 586:597 this complex 586:597 Although at least 7 of the 14 genes in this complex encode enzymatically active proteases, only one tryptase epsilon-like gene was identified. 11602603 1 109 theme data 195:198 arg1 base 200:203 sequence tag (EST) data base 176:203 sequence tag (EST) data base 176:203 Probing of the GenBank expressed sequence tag (EST) data base with varied human tryptase cDNAs identified two truncated ESTs that subsequently were found to encode overlapping portions of a novel human serine protease (designated tryptase epsilon or protease, serine S1 family member 22 (PRSS22)). 25826155 9 0 theme signal 1315:1320 arg1 mechanisms 1335:1344 signal transduction mechanisms 1315:1344 signal transduction mechanisms 1315:1344 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 2 1 from CTLR 363:366 arg1 cells 411:415 target cells 404:415 target cells 404:415 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 7 2 theme conformational 1041:1054 arg1 complementarity 1056:1070 the conformational complementarity 1037:1070 the conformational complementarity to explain a previous mutagenesis study 1037:1110 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 7 3 theme LLT1-NKRP1A 855:865 arg1 model 875:879 A LLT1-NKRP1A complex model 853:879 A LLT1-NKRP1A complex model 853:879 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 2 4 theme Th17 440:443 arg1 cells 445:449 control NK, NKT and Th17 cells 420:449 cells 445:449 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 7 5 theme complex 867:873 arg1 model 875:879 A LLT1-NKRP1A complex model 853:879 A LLT1-NKRP1A complex model 853:879 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 5 6 theme lectin-like 646:656 arg1 domain 658:663 the C-type lectin-like domain 635:663 the C-type lectin-like domain 635:663 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 9 7 theme structural 1255:1264 arg1 insights 1266:1273 structural insights 1255:1273 structural insights for understanding the binding modes 1255:1309 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 6 8 dep CTLR 769:772 arg1 keratinocyte-associated 775:797 keratinocyte-associated 775:797 keratinocyte-associated 775:797 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 3 9 theme structural 456:465 arg1 basis 467:471 The structural basis 452:471 The structural basis for the NKRP1A-LLT1 interaction 452:503 The structural basis for the NKRP1A-LLT1 interaction was limitedly understood. 25826155 3 10 theme NKRP1A-LLT1 481:491 arg1 interaction 493:503 the NKRP1A-LLT1 interaction 477:503 the NKRP1A-LLT1 interaction 477:503 The structural basis for the NKRP1A-LLT1 interaction was limitedly understood. 25826155 2 11 theme NKT 432:434 arg1 cells 445:449 control NK, NKT and Th17 cells 420:449 cells 445:449 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 9 12 dep provide 1247:1253 arg1 likely 1357:1362 likely 1357:1362 likely 1357:1362 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 9 13 theme further 1408:1414 arg1 design 1430:1435 further rational drug design 1408:1435 further rational drug design towards regulating the LLT1 function 1408:1472 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 8 14 theme crystal 1126:1132 arg1 packing 1134:1140 crystal packing 1126:1140 crystal packing 1126:1140 Furthermore, crystal packing and analytical ultracentrifugation revealed dimer formation, which supports a complex model. 25826155 5 15 theme extended 687:694 arg1 β-sheet 696:702 an extended β-sheet 684:702 an extended β-sheet 684:702 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 6 16 theme C-type 799:804 arg1 lectin 806:811 another CTLR, keratinocyte-associated C-type lectin 761:811 another CTLR, keratinocyte-associated C-type lectin 761:811 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 7 17 theme charge 1014:1019 arg1 consistency 1021:1031 the charge consistency 1010:1031 the charge consistency 1010:1031 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 2 18 theme CTLRs 334:338 arg1 CTLRs 334:338 the CTLRs 330:338 the CTLRs 330:338 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 18 theme CTLRs 334:338 arg1 one 323:325 one 323:325 one 323:325 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 19 theme natural 278:284 arg1 killer 286:291 Human natural killer 272:291 Human natural killer cell receptor-P1A (NKRP1A) 272:318 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 7 20 theme keratinocyte-associated 936:958 arg1 complex 980:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 1 21 theme pivotal 163:169 arg1 roles 171:175 the pivotal roles 159:175 the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses 159:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 0 22 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1) 0:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 0 22 theme Crystal 0:6 arg1 ligand 88:93 the ligand 84:93 the ligand for natural killer receptor-P1A 84:125 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 1 23 theme wide 240:243 arg1 range 245:249 a wide range 238:249 a wide range of immune responses 238:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 6 24 theme face 726:729 arg1 residues 709:716 The residues 705:716 The residues of this face 705:729 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 6 24 theme face 726:729 arg1 face 726:729 this face 721:729 this face 721:729 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 2 25 theme Human 272:276 arg1 killer 286:291 Human natural killer 272:291 Human natural killer cell receptor-P1A (NKRP1A) 272:318 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 4 26 theme ectodomain 576:585 arg1 structure 559:567 the crystal structure 547:567 the crystal structure of the ectodomain of LLT1 547:593 Here, we report the crystal structure of the ectodomain of LLT1. 25826155 0 27 theme extracellular 21:33 arg1 transcript 63:72 human lectin-like transcript 1 45:74 human lectin-like transcript 1 (LLT1) 45:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 0 27 theme extracellular 21:33 arg1 domain 35:40 extracellular domain 21:40 extracellular domain of human lectin-like transcript 1 (LLT1) 21:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 1 28 theme range 245:249 arg1 regulation 224:233 the regulation 220:233 the regulation of a wide range of immune responses 220:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 0 29 theme natural 99:105 arg1 killer 107:112 natural killer 99:112 natural killer receptor-P1A 99:125 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 9 30 theme CTLR 1387:1390 arg1 family 1392:1397 the CTLR family 1383:1397 the CTLR family 1383:1397 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 8 31 theme analytical 1146:1155 arg1 ultracentrifugation 1157:1175 analytical ultracentrifugation 1146:1175 analytical ultracentrifugation 1146:1175 Furthermore, crystal packing and analytical ultracentrifugation revealed dimer formation, which supports a complex model. 25826155 2 32 theme lectin-like 369:379 arg1 CTLR 363:366 another CTLR 355:366 another CTLR 355:366 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 32 theme lectin-like 369:379 arg1 transcript 381:390 lectin-like transcript 1 369:392 lectin-like transcript 1 (LLT1) 369:399 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 32 theme lectin-like 369:379 arg1 LLT1 395:398 LLT1 395:398 LLT1 395:398 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 7 33 theme crystal 901:907 arg1 structures 909:918 the crystal structures 897:918 the crystal structures of LLT1 897:926 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 1 34 theme C-type 180:185 arg1 CTLRs 210:214 CTLRs 210:214 CTLRs 210:214 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 1 34 theme C-type 180:185 arg1 receptors 199:207 C-type lectin-like receptors 180:207 C-type lectin-like receptors (CTLRs) 180:215 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 2 35 theme target 404:409 arg1 cells 411:415 target cells 404:415 target cells 404:415 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 1 36 theme immune 254:259 arg1 responses 261:269 immune responses 254:269 immune responses 254:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 7 37 theme C-type 960:965 arg1 complex 980:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 6 38 theme CTLR 769:772 arg1 lectin 806:811 another CTLR, keratinocyte-associated C-type lectin 761:811 another CTLR, keratinocyte-associated C-type lectin 761:811 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 1 39 theme lectin-like 187:197 arg1 CTLRs 210:214 CTLRs 210:214 CTLRs 210:214 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 1 39 theme lectin-like 187:197 arg1 receptors 199:207 C-type lectin-like receptors 180:207 C-type lectin-like receptors (CTLRs) 180:215 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 0 40 theme domain 35:40 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1) 0:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 0 40 theme domain 35:40 arg1 ligand 88:93 the ligand 84:93 the ligand for natural killer receptor-P1A 84:125 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 1 41 theme responses 261:269 arg1 range 245:249 a wide range 238:249 a wide range of immune responses 238:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 0 42 theme killer 107:112 arg1 receptor-P1A 114:125 natural killer receptor-P1A 99:125 natural killer receptor-P1A 99:125 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 9 43 theme drug 1425:1428 arg1 design 1430:1435 further rational drug design 1408:1435 further rational drug design towards regulating the LLT1 function 1408:1472 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 7 44 theme previous 1085:1092 arg1 study 1106:1110 a previous mutagenesis study 1083:1110 a previous mutagenesis study 1083:1110 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 1 45 theme receptors 199:207 arg1 roles 171:175 the pivotal roles 159:175 the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses 159:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 0 46 theme lectin-like 51:61 arg1 transcript 63:72 human lectin-like transcript 1 45:74 human lectin-like transcript 1 (LLT1) 45:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 0 46 theme lectin-like 51:61 arg1 LLT1 77:80 LLT1 77:80 LLT1 77:80 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 5 47 theme C-type 639:644 arg1 domain 658:663 the C-type lectin-like domain 635:663 the C-type lectin-like domain 635:663 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 2 48 theme control 420:426 arg1 cells 445:449 control NK, NKT and Th17 cells 420:449 cells 445:449 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 9 49 theme rational 1416:1423 arg1 design 1430:1435 further rational drug design 1408:1435 further rational drug design towards regulating the LLT1 function 1408:1472 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 7 50 theme lectin-NKp65 967:978 arg1 complex 980:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 0 51 theme human 45:49 arg1 transcript 63:72 human lectin-like transcript 1 45:74 human lectin-like transcript 1 (LLT1) 45:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 0 51 theme human 45:49 arg1 LLT1 77:80 LLT1 77:80 LLT1 77:80 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 5 52 theme receptor-binding 610:625 arg1 face 627:630 The plausible receptor-binding face 596:630 The plausible receptor-binding face of the C-type lectin-like domain 596:663 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 5 52 theme receptor-binding 610:625 arg1 flat 668:671 flat 668:671 flat 668:671 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 6 53 theme CTLR 833:836 arg1 NKp65 846:850 NKp65 846:850 NKp65 846:850 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 6 53 theme CTLR 833:836 arg1 member 838:843 the CTLR member 829:843 the CTLR member 829:843 The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. 25826155 7 54 theme LLT1 923:926 arg1 complex 980:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 the keratinocyte-associated C-type lectin-NKp65 complex 932:986 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 7 54 theme LLT1 923:926 arg1 structures 909:918 the crystal structures 897:918 the crystal structures of LLT1 897:926 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 4 55 theme LLT1 590:593 arg1 LLT1 590:593 LLT1 590:593 LLT1 590:593 Here, we report the crystal structure of the ectodomain of LLT1. 25826155 4 55 theme LLT1 590:593 arg1 ectodomain 576:585 the ectodomain 572:585 the ectodomain of LLT1 572:593 Here, we report the crystal structure of the ectodomain of LLT1. 25826155 2 56 theme cell 293:296 arg1 receptor-P1A 298:309 Human natural killer cell receptor-P1A 272:309 Human natural killer cell receptor-P1A (NKRP1A) 272:318 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 56 theme cell 293:296 arg1 NKRP1A 312:317 NKRP1A 312:317 NKRP1A 312:317 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 5 57 theme domain 658:663 arg1 face 627:630 The plausible receptor-binding face 596:630 The plausible receptor-binding face of the C-type lectin-like domain 596:663 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 5 57 theme domain 658:663 arg1 flat 668:671 flat 668:671 flat 668:671 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 5 58 theme plausible 600:608 arg1 face 627:630 The plausible receptor-binding face 596:630 The plausible receptor-binding face of the C-type lectin-like domain 596:663 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 5 58 theme plausible 600:608 arg1 flat 668:671 flat 668:671 flat 668:671 The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. 25826155 9 59 theme binding 1297:1303 arg1 modes 1305:1309 the binding modes 1293:1309 the binding modes 1293:1309 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 2 60 theme killer 286:291 arg1 receptor-P1A 298:309 Human natural killer cell receptor-P1A 272:309 Human natural killer cell receptor-P1A (NKRP1A) 272:318 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 60 theme killer 286:291 arg1 NKRP1A 312:317 NKRP1A 312:317 NKRP1A 312:317 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 2 61 theme NK 428:429 arg1 cells 445:449 control NK, NKT and Th17 cells 420:449 cells 445:449 Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. 25826155 4 62 theme crystal 551:557 arg1 structure 559:567 the crystal structure 547:567 the crystal structure of the ectodomain of LLT1 547:593 Here, we report the crystal structure of the ectodomain of LLT1. 25826155 8 63 theme complex 1220:1226 arg1 model 1228:1232 a complex model 1218:1232 a complex model 1218:1232 Furthermore, crystal packing and analytical ultracentrifugation revealed dimer formation, which supports a complex model. 25826155 1 64 from roles 171:175 arg1 regulation 224:233 the regulation 220:233 the regulation of a wide range of immune responses 220:269 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 9 65 theme LLT1 1460:1463 arg1 function 1465:1472 the LLT1 function 1456:1472 the LLT1 function 1456:1472 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 25826155 8 66 theme dimer 1186:1190 arg1 formation 1192:1200 dimer formation 1186:1200 dimer formation 1186:1200 Furthermore, crystal packing and analytical ultracentrifugation revealed dimer formation, which supports a complex model. 25826155 0 67 theme transcript 63:72 arg1 transcript 63:72 human lectin-like transcript 1 45:74 human lectin-like transcript 1 (LLT1) 45:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 0 67 theme transcript 63:72 arg1 domain 35:40 extracellular domain 21:40 extracellular domain of human lectin-like transcript 1 (LLT1) 21:81 Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A. 25826155 7 68 theme mutagenesis 1094:1104 arg1 study 1106:1110 a previous mutagenesis study 1083:1110 a previous mutagenesis study 1083:1110 A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. 25826155 1 69 theme Emerging 128:135 arg1 evidence 137:144 Emerging evidence 128:144 Emerging evidence 128:144 Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. 25826155 9 70 theme transduction 1322:1333 arg1 mechanisms 1335:1344 signal transduction mechanisms 1315:1344 signal transduction mechanisms 1315:1344 Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. 29666272 4 0 theme structural 929:938 arg1 basis 940:944 the structural basis 925:944 the structural basis for substrate recognition and catalysis 925:984 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 3 1 from core 747:750 arg1 branch 712:717 the second GlcNAcβ1,2- branch 689:717 the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor 689:786 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 2 2 theme branching 399:407 arg1 steps 417:421 Trimming and branching pathway steps 386:421 steps 417:421 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 10 3 theme structures 1970:1979 arg1 extension 1950:1958 regiospecific extension 1936:1958 regiospecific extension of glycan structures in mammalian cells 1936:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 3 4 theme sugar 776:780 arg1 donor 782:786 the sugar donor 772:786 the sugar donor 772:786 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 3 4 theme sugar 776:780 arg1 UDP-GlcNAc 758:767 UDP-GlcNAc 758:767 UDP-GlcNAc 758:767 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 3 5 theme glycans 596:602 arg1 synthesis 570:578 the synthesis 566:578 the synthesis of complex-type glycans 566:602 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 5 6 theme GT-A 1009:1012 arg1 fold 1028:1031 a GT-A Rossmann-like fold 1007:1031 a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor 1007:1129 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 10 7 theme mammalian 1984:1992 arg1 cells 1994:1998 mammalian cells 1984:1998 mammalian cells 1984:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 0 8 theme substrate 41:49 arg1 recognition 51:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. 29666272 8 9 theme glycan 1614:1619 arg1 pathway 1634:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 8 10 theme Recognition 1440:1450 arg1 interactions 1456:1467 Recognition arm interactions 1440:1467 Recognition arm interactions 1440:1467 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 9 11 theme subsite 1742:1748 arg1 modules 1750:1756 both conserved and convergent catalytic subsite modules 1702:1756 both conserved and convergent catalytic subsite modules 1702:1756 These data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. 29666272 7 12 theme exosite 1356:1362 arg1 pocket 1364:1369 a distal exosite pocket 1347:1369 a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1347:1436 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 10 13 from cells 1994:1998 arg1 extension 1950:1958 regiospecific extension 1936:1958 regiospecific extension of glycan structures in mammalian cells 1936:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 4 14 theme MGAT2 826:830 arg1 structure 807:815 the structure 803:815 the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis 803:984 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 0 15 theme convergent 107:116 arg1 exosite 118:124 a convergent exosite 105:124 a convergent exosite 105:124 Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. 29666272 10 16 theme MGAT2 1824:1828 arg1 architecture 1842:1853 the MGAT2 active-site architecture 1820:1853 the MGAT2 active-site architecture 1820:1853 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 7 17 theme Man-α1,6- 1309:1317 arg1 acceptor 1334:1341 the Man-α1,6- monosaccharide acceptor 1305:1341 the Man-α1,6- monosaccharide acceptor 1305:1341 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 5 18 theme divalent 1056:1063 arg1 interactions 1092:1103 conserved divalent cation-dependent substrate interactions 1046:1103 conserved divalent cation-dependent substrate interactions 1046:1103 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 8 19 theme Golgi 1522:1526 arg1 hydrolase 1558:1566 a glycoside hydrolase 1546:1566 a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway 1546:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 8 19 theme Golgi 1522:1526 arg1 α-mannosidase 1528:1540 Golgi α-mannosidase II 1522:1543 Golgi α-mannosidase II 1522:1543 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 0 20 theme Human 0:4 arg1 recognition 51:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. 29666272 9 21 theme catalytic 1732:1740 arg1 modules 1750:1756 both conserved and convergent catalytic subsite modules 1702:1756 both conserved and convergent catalytic subsite modules 1702:1756 These data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. 29666272 6 22 theme extended 1160:1167 arg1 acceptor 1176:1183 the extended glycan acceptor 1156:1183 the extended glycan acceptor 1156:1183 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 2 23 theme precise 470:476 arg1 specificities 488:500 the precise substrate specificities 466:500 the precise substrate specificities of the individual biosynthetic enzymes 466:539 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 0 24 theme II 38:39 arg1 recognition 51:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. 29666272 1 25 theme oligosaccharide 327:341 arg1 branches 343:350 multiple oligosaccharide branches 318:350 multiple oligosaccharide branches 318:350 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 10 26 theme glycan 1963:1968 arg1 structures 1970:1979 glycan structures 1963:1979 glycan structures in mammalian cells 1963:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 1 27 theme Asn-linked 127:136 arg1 oligosaccharides 138:153 Asn-linked oligosaccharides 127:153 Asn-linked oligosaccharides 127:153 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 5 28 theme UDP-GlcNAc 1114:1123 arg1 donor 1125:1129 the UDP-GlcNAc donor 1110:1129 the UDP-GlcNAc donor 1110:1129 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 6 29 theme MGAT2 1132:1136 arg1 interactions 1138:1149 MGAT2 interactions 1132:1149 MGAT2 interactions with the extended glycan acceptor 1132:1183 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 2 30 theme biosynthetic 520:531 arg1 enzymes 533:539 the individual biosynthetic enzymes 505:539 the individual biosynthetic enzymes 505:539 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 3 31 theme GlcNAcβ1,2- 700:710 arg1 branch 712:717 the second GlcNAcβ1,2- branch 689:717 the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor 689:786 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 4 32 theme GlcNAcMan3GlcNAc2-Asn 876:896 arg1 complex 907:913 a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis 874:984 a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis 874:984 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 6 33 from glycosyltransferases 1217:1236 arg1 distinct 1189:1196 distinct 1189:1196 distinct 1189:1196 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 3 34 theme committed 548:556 arg1 step 558:561 A key committed step 542:561 A key committed step in the synthesis of complex-type glycans 542:602 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 1 35 theme trimannosyl 361:371 arg1 core 380:383 the trimannosyl glycan core 357:383 the trimannosyl glycan core 357:383 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 6 36 with interactions 1138:1149 arg1 acceptor 1176:1183 the extended glycan acceptor 1156:1183 the extended glycan acceptor 1156:1183 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 7 37 theme recognition 1422:1432 arg1 arm 1434:1436 the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1382:1436 the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1382:1436 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 4 38 theme donor 846:850 arg1 complex 859:865 a Mn2+-UDP donor analog complex 835:865 a Mn2+-UDP donor analog complex 835:865 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 1 39 theme nascent 251:257 arg1 chains 266:271 the nascent glycan chains 247:271 the nascent glycan chains 247:271 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 5 40 theme substrate 1082:1090 arg1 interactions 1092:1103 conserved divalent cation-dependent substrate interactions 1046:1103 conserved divalent cation-dependent substrate interactions 1046:1103 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 7 41 theme substrate 1411:1419 arg1 arm 1434:1436 the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1382:1436 the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1382:1436 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 3 42 theme trimannosyl 728:738 arg1 core 747:750 the trimannosyl glycan core 724:750 the trimannosyl glycan core using UDP-GlcNAc as the sugar donor 724:786 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 1 43 theme chains 266:271 arg1 trimming 235:242 trimming 235:242 trimming of the nascent glycan chains 235:271 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 6 44 theme other 1203:1207 arg1 glycosyltransferases 1217:1236 other related glycosyltransferases 1203:1236 other related glycosyltransferases 1203:1236 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 2 45 theme pathway 409:415 arg1 steps 417:421 Trimming and branching pathway steps 386:421 steps 417:421 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 10 46 theme modular 1914:1920 arg1 templates 1922:1930 complementary modular templates 1900:1930 complementary modular templates for regiospecific extension of glycan structures in mammalian cells 1900:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 3 47 theme complex-type 583:594 arg1 glycans 596:602 complex-type glycans 583:602 complex-type glycans 583:602 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 1 48 theme secretory 207:215 arg1 pathway 217:223 the secretory pathway 203:223 the secretory pathway 203:223 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 4 49 theme acceptor 898:905 arg1 complex 907:913 a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis 874:984 a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis 874:984 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 10 50 from extension 1950:1958 arg1 cells 1994:1998 mammalian cells 1984:1998 mammalian cells 1984:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 3 51 from step 558:561 arg1 synthesis 570:578 the synthesis 566:578 the synthesis of complex-type glycans 566:602 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 2 52 theme Trimming 386:393 arg1 steps 417:421 Trimming and branching pathway steps 386:421 steps 417:421 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 8 53 theme Asn-linked 1603:1612 arg1 pathway 1634:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 4 54 theme substrate 950:958 arg1 recognition 960:970 substrate recognition 950:970 substrate recognition 950:970 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 8 55 theme biosynthetic 1621:1632 arg1 pathway 1634:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 7 56 theme distal 1349:1354 arg1 pocket 1364:1369 a distal exosite pocket 1347:1369 a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1347:1436 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 9 57 theme substrate 1667:1675 arg1 binding 1677:1683 substrate binding 1667:1683 substrate binding by MGAT2 1667:1692 These data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. 29666272 10 58 from structures 1970:1979 arg1 cells 1994:1998 mammalian cells 1984:1998 mammalian cells 1984:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 8 59 theme arm 1452:1454 arg1 interactions 1456:1467 Recognition arm interactions 1440:1467 Recognition arm interactions 1440:1467 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 7 60 theme monosaccharide 1319:1332 arg1 acceptor 1334:1341 the Man-α1,6- monosaccharide acceptor 1305:1341 the Man-α1,6- monosaccharide acceptor 1305:1341 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 4 61 theme human 820:824 arg1 MGAT2 826:830 human MGAT2 820:830 human MGAT2 820:830 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 5 62 theme conserved 1046:1054 arg1 interactions 1092:1103 conserved divalent cation-dependent substrate interactions 1046:1103 conserved divalent cation-dependent substrate interactions 1046:1103 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 8 63 link Asn-linked 1603:1612 arg1 pathway 1634:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 the Asn-linked glycan biosynthetic pathway 1599:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 8 64 theme enzyme-substrate 1488:1503 arg1 interactions 1505:1516 the enzyme-substrate interactions 1484:1516 the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway 1484:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 0 65 theme N-acetylglucosaminyltransferase 6:36 arg1 recognition 51:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition 0:61 Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. 29666272 9 66 theme convergent 1721:1730 arg1 modules 1750:1756 both conserved and convergent catalytic subsite modules 1702:1756 both conserved and convergent catalytic subsite modules 1702:1756 These data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. 29666272 6 67 theme glycan 1169:1174 arg1 acceptor 1176:1183 the extended glycan acceptor 1156:1183 the extended glycan acceptor 1156:1183 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 7 68 theme catalytic 1276:1284 arg1 subsite 1286:1292 a catalytic subsite 1274:1292 a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor 1274:1341 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 1 69 theme multiple 318:325 arg1 branches 343:350 multiple oligosaccharide branches 318:350 multiple oligosaccharide branches 318:350 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 5 70 theme Rossmann-like 1014:1026 arg1 fold 1028:1031 a GT-A Rossmann-like fold 1007:1031 a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor 1007:1129 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 2 71 theme substrate 478:486 arg1 specificities 488:500 the precise substrate specificities 466:500 the precise substrate specificities of the individual biosynthetic enzymes 466:539 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 9 72 theme conserved 1707:1715 arg1 modules 1750:1756 both conserved and convergent catalytic subsite modules 1702:1756 both conserved and convergent catalytic subsite modules 1702:1756 These data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. 29666272 2 73 theme enzymes 533:539 arg1 specificities 488:500 the precise substrate specificities 466:500 the precise substrate specificities of the individual biosynthetic enzymes 466:539 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 3 74 theme second 693:698 arg1 branch 712:717 the second GlcNAcβ1,2- branch 689:717 the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor 689:786 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 10 75 theme active-site 1830:1840 arg1 architecture 1842:1853 the MGAT2 active-site architecture 1820:1853 the MGAT2 active-site architecture 1820:1853 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 0 76 theme modular 70:76 arg1 architecture 78:89 a modular architecture 68:89 a modular architecture that includes a convergent exosite 68:124 Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite. 29666272 9 77 theme substrate 1769:1777 arg1 selectivity 1779:1789 substrate selectivity 1769:1789 substrate selectivity 1769:1789 These data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. 29666272 2 78 theme individual 509:518 arg1 enzymes 533:539 the individual biosynthetic enzymes 505:539 the individual biosynthetic enzymes 505:539 Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. 29666272 3 79 theme glycan 740:745 arg1 core 747:750 the trimannosyl glycan core 724:750 the trimannosyl glycan core using UDP-GlcNAc as the sugar donor 724:786 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 8 80 theme glycoside 1548:1556 arg1 hydrolase 1558:1566 a glycoside hydrolase 1546:1566 a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway 1546:1640 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 8 80 theme glycoside 1548:1556 arg1 α-mannosidase 1528:1540 Golgi α-mannosidase II 1522:1543 Golgi α-mannosidase II 1522:1543 Recognition arm interactions are similar to the enzyme-substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. 29666272 10 81 theme complementary 1900:1912 arg1 templates 1922:1930 complementary modular templates 1900:1930 complementary modular templates for regiospecific extension of glycan structures in mammalian cells 1900:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 29666272 1 82 theme glycan 373:378 arg1 core 380:383 the trimannosyl glycan core 357:383 the trimannosyl glycan core 357:383 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 5 83 theme cation-dependent 1065:1080 arg1 interactions 1092:1103 conserved divalent cation-dependent substrate interactions 1046:1103 conserved divalent cation-dependent substrate interactions 1046:1103 The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. 29666272 7 84 theme GlcNAc-β1,2Man-α1,3Manβ- 1386:1409 arg1 arm 1434:1436 the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1382:1436 the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm 1382:1436 These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate "recognition arm." 29666272 4 85 theme Mn2+-UDP 837:844 arg1 complex 859:865 a Mn2+-UDP donor analog complex 835:865 a Mn2+-UDP donor analog complex 835:865 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 3 86 theme key 544:546 arg1 step 558:561 A key committed step 542:561 A key committed step in the synthesis of complex-type glycans 542:602 A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. 29666272 1 87 theme glycan 259:264 arg1 chains 266:271 the nascent glycan chains 247:271 the nascent glycan chains 247:271 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 6 88 theme related 1209:1215 arg1 glycosyltransferases 1217:1236 other related glycosyltransferases 1203:1236 other related glycosyltransferases 1203:1236 MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. 29666272 1 89 link Asn-linked 127:136 arg1 oligosaccharides 138:153 Asn-linked oligosaccharides 127:153 Asn-linked oligosaccharides 127:153 Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. 29666272 4 90 theme analog 852:857 arg1 complex 859:865 a Mn2+-UDP donor analog complex 835:865 a Mn2+-UDP donor analog complex 835:865 We determined the structure of human MGAT2 as a Mn2+-UDP donor analog complex and as a GlcNAcMan3GlcNAc2-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. 29666272 10 91 theme regiospecific 1936:1948 arg1 extension 1950:1958 regiospecific extension 1936:1958 regiospecific extension of glycan structures in mammalian cells 1936:1998 More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells. 21606496 0 0 theme cell 83:86 arg1 2B4 97:99 the activating natural killer cell receptor 2B4 53:99 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 0 0 theme cell 83:86 arg1 CD244 102:106 CD244 102:106 CD244 102:106 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 7 1 theme important 1032:1040 arg1 impact 1042:1047 an important impact 1029:1047 an important impact 1029:1047 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 2 2 theme NK 306:307 arg1 cells 309:313 primary human NK cells 292:313 primary human NK cells 292:313 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 7 3 contain has 1025:1027 arg2 impact 1042:1047 an important impact 1029:1047 an important impact 1029:1047 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 7 3 contain has 1025:1027 arg1 glycosylation 1011:1023 glycosylation 1011:1023 glycosylation 1011:1023 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 6 4 theme glycosylation 889:901 arg1 desialylation 833:845 the desialylation 829:845 the desialylation of NK cells 829:857 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 6 4 theme glycosylation 889:901 arg1 inhibition 866:875 the inhibition 862:875 the inhibition of O-linked glycosylation 862:901 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 0 5 theme killer 76:81 arg1 2B4 97:99 the activating natural killer cell receptor 2B4 53:99 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 0 5 theme killer 76:81 arg1 CD244 102:106 CD244 102:106 CD244 102:106 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 4 6 theme fusion 463:468 arg1 protein 470:476 a recombinant fusion protein 449:476 a recombinant fusion protein of the extracellular domain of 2B4 449:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 4 7 gly glycosylation 543:555 arg1 2B4 560:562 2B4 560:562 2B4 560:562 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 2 8 theme human 300:304 arg1 cells 309:313 primary human NK cells 292:313 primary human NK cells 292:313 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 1 9 theme cell 208:211 arg1 responses 213:221 natural killer (NK) cell responses 188:221 natural killer (NK) cell responses 188:221 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 0 10 theme 2B4 97:99 arg1 protein 109:115 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 4 11 theme N-linked 534:541 arg1 glycosylation 543:555 N-linked glycosylation 534:555 N-linked glycosylation of 2B4 534:562 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 1 12 theme responses 213:221 arg1 regulation 174:183 the regulation 170:183 the regulation of natural killer (NK) cell responses 170:221 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 0 13 theme receptor 88:95 arg1 2B4 97:99 the activating natural killer cell receptor 2B4 53:99 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 0 13 theme receptor 88:95 arg1 CD244 102:106 CD244 102:106 CD244 102:106 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 7 14 theme regulated 1099:1107 arg1 changes 1109:1115 regulated changes 1099:1115 regulated changes in glycosylation during NK cell development and activation 1099:1174 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 7 15 theme responses 1223:1231 arg1 regulation 1201:1210 the regulation 1197:1210 the regulation of NK cell responses 1197:1231 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 1 16 theme activating 146:155 arg1 receptor 157:164 an important activating receptor 133:164 an important activating receptor for the regulation of natural killer (NK) cell responses 133:221 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 1 16 theme activating 146:155 arg1 2B4 118:120 2B4 118:120 2B4 (CD244) 118:128 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 5 17 theme sialic 760:765 arg1 acids 767:771 sialic acids 760:771 sialic acids 760:771 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 7 18 theme 2B4-mediated 1052:1063 arg1 function 1073:1080 2B4-mediated NK cell function 1052:1080 2B4-mediated NK cell function 1052:1080 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 6 19 theme increased 915:923 arg1 lysis 938:942 increased 2B4-mediated lysis 915:942 increased 2B4-mediated lysis of CD48-expressing tumor target cells 915:980 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 1 20 theme important 136:144 arg1 receptor 157:164 an important activating receptor 133:164 an important activating receptor for the regulation of natural killer (NK) cell responses 133:221 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 1 20 theme important 136:144 arg1 2B4 118:120 2B4 118:120 2B4 (CD244) 118:128 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 6 21 theme NK 850:851 arg1 cells 853:857 NK cells 850:857 NK cells 850:857 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 7 22 theme NK 1141:1142 arg1 development 1149:1159 NK cell development 1141:1159 NK cell development 1141:1159 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 5 23 theme ligand 670:675 arg1 binding 677:683 ligand binding 670:683 ligand binding 670:683 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 2 24 theme cell 322:325 arg1 lines 327:331 NK cell lines 319:331 NK cell lines 319:331 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 6 25 theme assay 809:813 arg1 system 815:820 a functional assay system 796:820 a functional assay system 796:820 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 5 26 contain has 645:647 arg1 sialylation 626:636 sialylation 626:636 sialylation of 2B4 626:643 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 5 26 contain has 645:647 arg2 impact 660:665 a negative impact 649:665 a negative impact 649:665 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 5 27 theme acids 767:771 arg1 removal 749:755 the removal 745:755 the removal of sialic acids 745:771 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 2 28 theme NK 319:320 arg1 lines 327:331 NK cell lines 319:331 NK cell lines 319:331 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 4 29 theme domain 499:504 arg1 protein 470:476 a recombinant fusion protein 449:476 a recombinant fusion protein of the extracellular domain of 2B4 449:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 0 30 theme protein 109:115 arg1 function 41:48 function 41:48 function 41:48 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 0 30 theme protein 109:115 arg1 binding 29:35 ligand binding 22:35 ligand binding 22:35 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 6 31 gly desialylation 833:845 arg1 cells 853:857 NK cells 850:857 NK cells 850:857 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 6 31 gly desialylation 833:845 arg1 glycosylation 889:901 O-linked glycosylation 880:901 O-linked glycosylation 880:901 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 6 32 theme tumor 963:967 arg1 cells 976:980 CD48-expressing tumor target cells 947:980 CD48-expressing tumor target cells 947:980 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 0 33 theme ligand 22:27 arg1 binding 29:35 ligand binding 22:35 ligand binding 22:35 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 6 34 theme cells 853:857 arg1 desialylation 833:845 the desialylation 829:845 the desialylation of NK cells 829:857 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 6 34 theme cells 853:857 arg1 inhibition 866:875 the inhibition 862:875 the inhibition of O-linked glycosylation 862:901 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 6 35 link O-linked 880:887 arg1 glycosylation 889:901 O-linked glycosylation 880:901 O-linked glycosylation 880:901 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 6 36 theme CD48-expressing 947:961 arg1 cells 976:980 CD48-expressing tumor target cells 947:980 CD48-expressing tumor target cells 947:980 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 7 37 theme NK 1065:1066 arg1 function 1073:1080 2B4-mediated NK cell function 1052:1080 2B4-mediated NK cell function 1052:1080 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 4 38 theme 2B4 509:511 arg1 2B4 509:511 2B4 509:511 2B4 509:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 4 38 theme 2B4 509:511 arg1 domain 499:504 the extracellular domain 481:504 the extracellular domain of 2B4 481:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 5 39 theme 2B4 641:643 arg1 sialylation 626:636 sialylation 626:636 sialylation of 2B4 626:643 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 7 40 from changes 1109:1115 arg1 glycosylation 1120:1132 glycosylation 1120:1132 glycosylation 1120:1132 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 4 41 theme ligand 600:605 arg1 CD48 607:610 its ligand CD48 596:610 its ligand CD48 596:610 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 3 42 theme acid 395:398 arg1 residues 400:407 sialic acid residues 388:407 sialic acid residues on N- and O-linked carbohydrates 388:440 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 7 43 theme cell 1068:1071 arg1 function 1073:1080 2B4-mediated NK cell function 1052:1080 2B4-mediated NK cell function 1052:1080 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 6 44 theme O-linked 880:887 arg1 glycosylation 889:901 O-linked glycosylation 880:901 O-linked glycosylation 880:901 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 5 45 theme negative 651:658 arg1 impact 660:665 a negative impact 649:665 a negative impact 649:665 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 6 46 theme cells 976:980 arg1 lysis 938:942 increased 2B4-mediated lysis 915:942 increased 2B4-mediated lysis of CD48-expressing tumor target cells 915:980 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 7 47 theme cell 1144:1147 arg1 development 1149:1159 NK cell development 1141:1159 NK cell development 1141:1159 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 3 48 from residues 400:407 arg1 carbohydrates 428:440 O-linked carbohydrates 419:440 O-linked carbohydrates 419:440 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 3 48 from residues 400:407 arg1 N- 412:413 N- 412:413 N- 412:413 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 2 49 theme primary 292:298 arg1 cells 309:313 primary human NK cells 292:313 primary human NK cells 292:313 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 3 50 theme differential 338:349 arg1 glycosylation 351:363 The differential glycosylation 334:363 The differential glycosylation 334:363 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 4 51 theme extracellular 485:497 arg1 2B4 509:511 2B4 509:511 2B4 509:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 4 51 theme extracellular 485:497 arg1 domain 499:504 the extracellular domain 481:504 the extracellular domain of 2B4 481:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 3 52 link O-linked 419:426 arg1 carbohydrates 428:440 O-linked carbohydrates 419:440 O-linked carbohydrates 419:440 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 4 53 link N-linked 534:541 arg1 glycosylation 543:555 N-linked glycosylation 534:555 N-linked glycosylation of 2B4 534:562 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 5 54 gly sialylation 626:636 arg1 2B4 641:643 2B4 641:643 2B4 641:643 In contrast, sialylation of 2B4 has a negative impact on ligand binding, as the interaction between 2B4 and CD48 is increased after the removal of sialic acids. 21606496 1 55 theme natural 188:194 arg1 NK 204:205 NK 204:205 NK 204:205 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 1 55 theme natural 188:194 arg1 killer 196:201 natural killer 188:201 natural killer (NK) cell responses 188:221 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 2 56 gly glycosylated 276:287 arg1 lines 327:331 NK cell lines 319:331 NK cell lines 319:331 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 2 56 gly glycosylated 276:287 arg1 cells 309:313 primary human NK cells 292:313 primary human NK cells 292:313 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 2 56 gly glycosylated 276:287 arg2 2B4 242:244 2B4 242:244 2B4 242:244 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 2 56 gly glycosylated 276:287 arg1 2B4 242:244 2B4 242:244 2B4 242:244 Here we show that 2B4 is heavily and differentially glycosylated in primary human NK cells and NK cell lines. 21606496 4 57 theme recombinant 451:461 arg1 protein 470:476 a recombinant fusion protein 449:476 a recombinant fusion protein of the extracellular domain of 2B4 449:511 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 21606496 6 58 theme target 969:974 arg1 cells 976:980 CD48-expressing tumor target cells 947:980 CD48-expressing tumor target cells 947:980 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 3 59 theme O-linked 419:426 arg1 carbohydrates 428:440 O-linked carbohydrates 419:440 O-linked carbohydrates 419:440 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 1 60 theme killer 196:201 arg1 responses 213:221 natural killer (NK) cell responses 188:221 natural killer (NK) cell responses 188:221 2B4 (CD244) is an important activating receptor for the regulation of natural killer (NK) cell responses. 21606496 0 61 theme natural 68:74 arg1 killer 76:81 the activating natural killer 53:81 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 7 62 theme NK 1215:1216 arg1 responses 1223:1231 NK cell responses 1215:1231 NK cell responses 1215:1231 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 7 63 theme cell 1218:1221 arg1 responses 1223:1231 NK cell responses 1215:1231 NK cell responses 1215:1231 These data demonstrate that glycosylation has an important impact on 2B4-mediated NK cell function and suggest that regulated changes in glycosylation during NK cell development and activation might be involved in the regulation of NK cell responses. 21606496 3 64 theme sialic 388:393 arg1 residues 400:407 sialic acid residues 388:407 sialic acid residues on N- and O-linked carbohydrates 388:440 The differential glycosylation could be attributed to sialic acid residues on N- and O-linked carbohydrates. 21606496 6 65 theme functional 798:807 arg1 system 815:820 a functional assay system 796:820 a functional assay system 796:820 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 0 66 theme activating 57:66 arg1 killer 76:81 the activating natural killer 53:81 the activating natural killer cell receptor 2B4 (CD244) protein 53:115 Glycosylation affects ligand binding and function of the activating natural killer cell receptor 2B4 (CD244) protein. 21606496 6 67 theme 2B4-mediated 925:936 arg1 lysis 938:942 increased 2B4-mediated lysis 915:942 increased 2B4-mediated lysis of CD48-expressing tumor target cells 915:980 This was confirmed in a functional assay system, where the desialylation of NK cells or the inhibition of O-linked glycosylation resulted in increased 2B4-mediated lysis of CD48-expressing tumor target cells. 21606496 4 68 theme 2B4 560:562 arg1 glycosylation 543:555 N-linked glycosylation 534:555 N-linked glycosylation of 2B4 534:562 Using a recombinant fusion protein of the extracellular domain of 2B4, we demonstrate that N-linked glycosylation of 2B4 is essential for the binding to its ligand CD48. 27152332 6 0 theme other 1200:1204 arg1 structure 1220:1228 any other known protein structure 1196:1228 any other known protein structure 1196:1228 The dimerization domain adopts a conformation that bears no resemblance to any other known protein structure. 27152332 6 1 theme known 1206:1210 arg1 structure 1220:1228 any other known protein structure 1196:1228 any other known protein structure 1196:1228 The dimerization domain adopts a conformation that bears no resemblance to any other known protein structure. 27152332 5 2 theme copper 1108:1113 arg1 what 1074:1077 what 1074:1077 what 1074:1077 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 2 theme copper 1108:1113 arg1 sites 1040:1044 the two copper-binding sites 1017:1044 the two copper-binding sites 1017:1044 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 2 theme copper 1108:1113 arg1 site 1115:1118 a coupled binuclear copper site 1088:1118 a coupled binuclear copper site 1088:1118 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 3 3 theme crystal 477:483 arg1 structure 485:493 the crystal structure 473:493 the crystal structure 473:493 We report the crystal structure of human dopamine β-hydroxylase, which is the enzyme converting dopamine to norepinephrine. 27152332 3 3 theme crystal 477:483 arg1 enzyme 541:546 the enzyme 537:546 the enzyme converting dopamine to norepinephrine 537:584 We report the crystal structure of human dopamine β-hydroxylase, which is the enzyme converting dopamine to norepinephrine. 27152332 7 4 theme numerous 1286:1293 arg1 disorders 1307:1315 the numerous devastating disorders 1282:1315 the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system 1282:1398 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 3 5 theme β-hydroxylase 513:525 arg1 structure 485:493 the crystal structure 473:493 the crystal structure 473:493 We report the crystal structure of human dopamine β-hydroxylase, which is the enzyme converting dopamine to norepinephrine. 27152332 3 5 theme β-hydroxylase 513:525 arg1 enzyme 541:546 the enzyme 537:546 the enzyme converting dopamine to norepinephrine 537:584 We report the crystal structure of human dopamine β-hydroxylase, which is the enzyme converting dopamine to norepinephrine. 27152332 4 6 theme possible 706:713 arg1 site 729:732 a possible metal-binding site 704:732 a possible metal-binding site 704:732 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 5 7 theme α-hydroxylating 922:936 arg1 monooxygenase 956:968 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 2 8 theme example 297:303 arg1 hypertension 305:316 example hypertension 297:316 example hypertension 297:316 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 5 9 theme core 777:780 arg1 structure 782:790 The catalytic core structure 763:790 The catalytic core structure 763:790 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 3 10 theme human 498:502 arg1 β-hydroxylase 513:525 human dopamine β-hydroxylase 498:525 human dopamine β-hydroxylase 498:525 We report the crystal structure of human dopamine β-hydroxylase, which is the enzyme converting dopamine to norepinephrine. 27152332 5 11 theme open 830:833 arg1 site 842:845 an open active site 827:845 two different conformations: an open active site 798:845 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 12 dep structure 997:1005 arg1 what 1074:1077 what 1074:1077 what 1074:1077 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 12 dep structure 997:1005 arg1 sites 1040:1044 the two copper-binding sites 1017:1044 the two copper-binding sites 1017:1044 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 12 dep structure 997:1005 arg1 site 1115:1118 a coupled binuclear copper site 1088:1118 a coupled binuclear copper site 1088:1118 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 4 13 located found 665:669 arg2 structure 591:599 The structure 587:599 The structure 587:599 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 4 13 located found 665:669 arg1 proteins 686:693 >1600 other proteins 674:693 >1600 other proteins 674:693 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 0 14 theme dopamine 31:38 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human dopamine 0:38 The crystal structure of human dopamine β-hydroxylase at 2.9 Å resolution. 27152332 3 15 theme dopamine 504:511 arg1 β-hydroxylase 513:525 human dopamine β-hydroxylase 498:525 human dopamine β-hydroxylase 498:525 We report the crystal structure of human dopamine β-hydroxylase, which is the enzyme converting dopamine to norepinephrine. 27152332 5 16 theme active 835:840 arg1 site 842:845 an open active site 827:845 two different conformations: an open active site 798:845 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 2 17 theme hyperactivity 415:427 arg1 disorder 429:436 attention deficit hyperactivity disorder 397:436 attention deficit hyperactivity disorder 397:436 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 5 18 dep 5 1060:1060 arg1 to 1057:1058 to 1057:1058 to 1057:1058 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 19 theme enzyme 887:892 arg1 monooxygenase 956:968 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 4 20 theme domain 652:657 arg1 structure 591:599 The structure 587:599 The structure 587:599 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 2 21 theme deficit 407:413 arg1 disorder 429:436 attention deficit hyperactivity disorder 397:436 attention deficit hyperactivity disorder 397:436 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 1 22 theme norepinephrine 79:92 arg1 pathway 94:100 The norepinephrine pathway 75:100 The norepinephrine pathway 75:100 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 2 23 theme attention 397:405 arg1 disorder 429:436 attention deficit hyperactivity disorder 397:436 attention deficit hyperactivity disorder 397:436 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 7 24 theme neurological 1343:1354 arg1 origins 1356:1362 both physiological and neurological origins 1320:1362 both physiological and neurological origins 1320:1362 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 5 25 theme active 985:990 arg1 structure 997:1005 a closed active site structure 976:1005 a closed active site structure 976:1005 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 4 26 theme metal-binding 715:727 arg1 site 729:732 a possible metal-binding site 704:732 a possible metal-binding site 704:732 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 2 27 from abnormalities 225:237 arg1 pathway 246:252 the pathway 242:252 the pathway 242:252 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 4 28 theme DOMON 608:612 arg1 domain 652:657 the DOMON (dopamine β-monooxygenase N-terminal) domain 604:657 the DOMON (dopamine β-monooxygenase N-terminal) domain 604:657 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 5 29 theme family 894:899 arg1 monooxygenase 956:968 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 30 theme coupled 1090:1096 arg1 what 1074:1077 what 1074:1077 what 1074:1077 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 30 theme coupled 1090:1096 arg1 sites 1040:1044 the two copper-binding sites 1017:1044 the two copper-binding sites 1017:1044 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 30 theme coupled 1090:1096 arg1 site 1115:1118 a coupled binuclear copper site 1088:1118 a coupled binuclear copper site 1088:1118 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 4 31 theme ligand-binding 740:753 arg1 pocket 755:760 a ligand-binding pocket 738:760 a ligand-binding pocket 738:760 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 2 32 theme numerous 274:281 arg1 diseases 283:290 numerous diseases 274:290 numerous diseases 274:290 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 6 33 contain bears 1172:1176 arg1 conformation 1154:1165 a conformation 1152:1165 a conformation that bears no resemblance to any other known protein structure 1152:1228 The dimerization domain adopts a conformation that bears no resemblance to any other known protein structure. 27152332 6 33 contain bears 1172:1176 arg2 resemblance 1181:1191 no resemblance 1178:1191 no resemblance to any other known protein structure 1178:1228 The dimerization domain adopts a conformation that bears no resemblance to any other known protein structure. 27152332 2 34 theme cocaine 443:449 arg1 dependence 451:460 cocaine dependence 443:460 cocaine dependence 443:460 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 5 35 theme binuclear 1098:1106 arg1 what 1074:1077 what 1074:1077 what 1074:1077 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 35 theme binuclear 1098:1106 arg1 sites 1040:1044 the two copper-binding sites 1017:1044 the two copper-binding sites 1017:1044 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 35 theme binuclear 1098:1106 arg1 site 1115:1118 a coupled binuclear copper site 1088:1118 a coupled binuclear copper site 1088:1118 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 0 36 theme human 25:29 arg1 dopamine 31:38 human dopamine 25:38 human dopamine 25:38 The crystal structure of human dopamine β-hydroxylase at 2.9 Å resolution. 27152332 2 37 attach linked 264:269 arg1 diseases 283:290 numerous diseases 274:290 numerous diseases 274:290 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 2 37 attach linked 264:269 arg2 abnormalities 225:237 abnormalities 225:237 abnormalities in the pathway 225:252 Furthermore, abnormalities in the pathway have been linked to numerous diseases, for example hypertension, depression, anxiety, Parkinson's disease, schizophrenia, Alzheimer's disease, attention deficit hyperactivity disorder, and cocaine dependence. 27152332 5 38 from site 1115:1118 arg1 which 1011:1015 which 1011:1015 which 1011:1015 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 7 39 theme devastating 1295:1305 arg1 disorders 1307:1315 the numerous devastating disorders 1282:1315 the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system 1282:1398 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 5 40 theme copper-binding 1025:1038 arg1 what 1074:1077 what 1074:1077 what 1074:1077 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 40 theme copper-binding 1025:1038 arg1 sites 1040:1044 the two copper-binding sites 1017:1044 the two copper-binding sites 1017:1044 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 40 theme copper-binding 1025:1038 arg1 site 1115:1118 a coupled binuclear copper site 1088:1118 a coupled binuclear copper site 1088:1118 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 41 theme closed 978:983 arg1 structure 997:1005 a closed active site structure 976:1005 a closed active site structure 976:1005 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 7 42 theme dopamine 1384:1391 arg1 system 1393:1398 the dopamine system 1380:1398 the dopamine system 1380:1398 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 4 43 theme other 680:684 arg1 proteins 686:693 >1600 other proteins 674:693 >1600 other proteins 674:693 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 5 44 theme catalytic 767:775 arg1 structure 782:790 The catalytic core structure 763:790 The catalytic core structure 763:790 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 45 theme site 992:995 arg1 structure 997:1005 a closed active site structure 976:1005 a closed active site structure 976:1005 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 1 46 theme physiological 141:153 arg1 mood 174:177 mood 174:177 mood 174:177 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 1 46 theme physiological 141:153 arg1 arousal 188:194 arousal 188:194 arousal 188:194 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 1 46 theme physiological 141:153 arg1 attention 201:209 attention 201:209 attention 201:209 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 1 46 theme physiological 141:153 arg1 processes 155:163 behavioral and physiological processes 126:163 behavioral and physiological processes 126:163 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 6 47 theme dimerization 1125:1136 arg1 domain 1138:1143 The dimerization domain 1121:1143 The dimerization domain 1121:1143 The dimerization domain adopts a conformation that bears no resemblance to any other known protein structure. 27152332 6 48 theme protein 1212:1218 arg1 structure 1220:1228 any other known protein structure 1196:1228 any other known protein structure 1196:1228 The dimerization domain adopts a conformation that bears no resemblance to any other known protein structure. 27152332 4 49 dep domain 652:657 arg1 N-terminal 640:649 N-terminal 640:649 N-terminal 640:649 The structure of the DOMON (dopamine β-monooxygenase N-terminal) domain, also found in >1600 other proteins, reveals a possible metal-binding site and a ligand-binding pocket. 27152332 7 50 theme origins 1356:1362 arg1 disorders 1307:1315 the numerous devastating disorders 1282:1315 the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system 1282:1398 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 5 51 theme [the 901:904 arg1 monooxygenase 956:968 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 7 52 theme physiological 1325:1337 arg1 origins 1356:1362 both physiological and neurological origins 1320:1362 both physiological and neurological origins 1320:1362 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 5 53 theme α-amidating 943:953 arg1 monooxygenase 956:968 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 54 theme monooxygenase 956:968 arg1 member 872:877 another member 864:877 another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 864:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 1 55 theme behavioral 126:135 arg1 mood 174:177 mood 174:177 mood 174:177 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 1 55 theme behavioral 126:135 arg1 arousal 188:194 arousal 188:194 arousal 188:194 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 1 55 theme behavioral 126:135 arg1 attention 201:209 attention 201:209 attention 201:209 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 1 55 theme behavioral 126:135 arg1 processes 155:163 behavioral and physiological processes 126:163 behavioral and physiological processes 126:163 The norepinephrine pathway is believed to modulate behavioral and physiological processes, such as mood, overall arousal, and attention. 27152332 0 56 theme Å 61:61 arg1 resolution 63:72 2.9 Å resolution 57:72 2.9 Å resolution 57:72 The crystal structure of human dopamine β-hydroxylase at 2.9 Å resolution. 27152332 7 57 theme molecular 1258:1266 arg1 insights 1268:1275 new molecular insights 1254:1275 new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system 1254:1398 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 5 58 theme different 802:810 arg1 conformations 812:824 two different conformations 798:824 two different conformations: an open active site 798:845 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 5 59 theme peptidylglycine 906:920 arg1 monooxygenase 956:968 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase] 882:969 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 7 60 theme new 1254:1256 arg1 insights 1268:1275 new molecular insights 1254:1275 new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system 1254:1398 The structure provides new molecular insights into the numerous devastating disorders of both physiological and neurological origins associated with the dopamine system. 27152332 5 61 dep conformations 812:824 arg1 site 842:845 an open active site 827:845 two different conformations: an open active site 798:845 The catalytic core structure shows two different conformations: an open active site, as also seen in another member of this enzyme family [the peptidylglycine α-hydroxylating (and α-amidating) monooxygenase], and a closed active site structure, in which the two copper-binding sites are only 4 to 5 Å apart, in what might be a coupled binuclear copper site. 27152332 0 62 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human dopamine 0:38 The crystal structure of human dopamine β-hydroxylase at 2.9 Å resolution. 23035744 0 0 from Structures 0:9 arg1 complex 44:50 complex 44:50 complex with pharmacologically important ligands 44:91 Structures of human acetylcholinesterase in complex with pharmacologically important ligands. 23035744 2 1 theme disease 309:315 arg1 donepezil 283:291 donepezil 283:291 donepezil 283:291 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 2 1 theme disease 309:315 arg1 drug 317:320 an Alzheimer's disease drug 294:320 an Alzheimer's disease drug 294:320 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 3 2 theme human 403:407 arg1 AChE 409:412 human AChE 403:412 human AChE 403:412 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 1 3 theme Human 94:98 arg1 target 145:150 a significant target 131:150 a significant target for therapeutic drugs 131:172 Human acetylcholinesterase (AChE) is a significant target for therapeutic drugs. 23035744 1 3 theme Human 94:98 arg1 AChE 122:125 AChE 122:125 AChE 122:125 Human acetylcholinesterase (AChE) is a significant target for therapeutic drugs. 23035744 1 3 theme Human 94:98 arg1 acetylcholinesterase 100:119 Human acetylcholinesterase 94:119 Human acetylcholinesterase (AChE) 94:126 Human acetylcholinesterase (AChE) is a significant target for therapeutic drugs. 23035744 3 4 theme AChE 409:412 arg1 crystals 391:398 These crystals 385:398 These crystals of human AChE 385:412 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 0 5 theme acetylcholinesterase 20:39 arg1 Structures 0:9 Structures 0:9 Structures of human acetylcholinesterase in complex with pharmacologically important ligands 0:91 Structures of human acetylcholinesterase in complex with pharmacologically important ligands. 23035744 0 6 theme human 14:18 arg1 acetylcholinesterase 20:39 human acetylcholinesterase 14:39 human acetylcholinesterase 14:39 Structures of human acetylcholinesterase in complex with pharmacologically important ligands. 23035744 2 7 theme high 191:194 arg1 structures 215:224 high resolution crystal structures 191:224 high resolution crystal structures of human AChE 191:238 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 3 8 dep available 492:500 arg1 than 476:479 than 476:479 than 476:479 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 3 9 theme accurate 429:436 arg1 platform 438:445 a more accurate platform 422:445 a more accurate platform for further drug development than previously available 422:500 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 2 10 theme AChE 235:238 arg1 structures 215:224 high resolution crystal structures 191:224 high resolution crystal structures of human AChE 191:238 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 3 11 theme further 451:457 arg1 development 464:474 further drug development 451:474 further drug development than previously available 451:500 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 2 12 theme human 229:233 arg1 AChE 235:238 human AChE 229:238 human AChE 229:238 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 3 13 theme drug 459:462 arg1 development 464:474 further drug development 451:474 further drug development than previously available 451:500 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 2 14 with complexes 254:262 arg1 ligands 274:280 drug ligands 269:280 drug ligands 269:280 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 1 15 theme significant 133:143 arg1 target 145:150 a significant target 131:150 a significant target for therapeutic drugs 131:172 Human acetylcholinesterase (AChE) is a significant target for therapeutic drugs. 23035744 1 15 theme significant 133:143 arg1 acetylcholinesterase 100:119 Human acetylcholinesterase 94:119 Human acetylcholinesterase (AChE) 94:126 Human acetylcholinesterase (AChE) is a significant target for therapeutic drugs. 23035744 2 16 theme drug 269:272 arg1 ligands 274:280 drug ligands 269:280 drug ligands 269:280 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 2 17 theme crystal 207:213 arg1 structures 215:224 high resolution crystal structures 191:224 high resolution crystal structures of human AChE 191:238 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 3 18 theme available 492:500 arg1 development 464:474 further drug development 451:474 further drug development than previously available 451:500 These crystals of human AChE provide a more accurate platform for further drug development than previously available. 23035744 2 19 theme resolution 196:205 arg1 structures 215:224 high resolution crystal structures 191:224 high resolution crystal structures of human AChE 191:238 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 23035744 1 20 theme therapeutic 156:166 arg1 drugs 168:172 therapeutic drugs 156:172 therapeutic drugs 156:172 Human acetylcholinesterase (AChE) is a significant target for therapeutic drugs. 23035744 0 21 theme important 75:83 arg1 ligands 85:91 pharmacologically important ligands 57:91 pharmacologically important ligands 57:91 Structures of human acetylcholinesterase in complex with pharmacologically important ligands. 23035744 0 22 with complex 44:50 arg1 ligands 85:91 pharmacologically important ligands 57:91 pharmacologically important ligands 57:91 Structures of human acetylcholinesterase in complex with pharmacologically important ligands. 23035744 2 23 theme human 344:348 arg1 AChE 350:353 human AChE than it does to Torpedo AChE 344:382 human AChE than it does to Torpedo AChE 344:382 Here we present high resolution crystal structures of human AChE, alone and in complexes with drug ligands; donepezil, an Alzheimer's disease drug, binds differently to human AChE than it does to Torpedo AChE. 2141278 0 0 theme epidermal 78:86 arg1 factor 95:100 epidermal growth factor 78:100 the epidermal growth factor homology region 74:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 4 1 used occupied 1085:1092 arg2 position 1061:1068 position 134 1061:1072 position 134 of C1-s 1061:1080 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 0 2 contain containing 63:72 arg2 region 111:116 the epidermal growth factor homology region 74:116 the epidermal growth factor homology region 74:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 0 2 contain containing 63:72 arg1 C1-s 58:61 C1-s 58:61 C1-s containing the epidermal growth factor homology region 58:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 7 3 theme C1-s-C1r-C1-r-C1-s 1753:1770 arg1 tetramer 1772:1779 the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer 1732:1779 the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer 1732:1779 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 3 4 theme epidermal 678:686 arg1 region 718:723 the epidermal growth factor-like (EGF-like) region 674:723 the epidermal growth factor-like (EGF-like) region of C1-s 674:731 Fragment alpha 2, which comprised the epidermal growth factor-like (EGF-like) region of C1-s, was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195. 2141278 1 5 theme subcomponents 142:154 arg1 one 125:127 one 125:127 one 125:127 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 5 theme subcomponents 142:154 arg1 C1-s 119:122 C1-s 119:122 C1-s 119:122 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 5 theme subcomponents 142:154 arg1 subcomponents 142:154 the three subcomponents 132:154 the three subcomponents of C1-, the first component of complement 132:196 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 5 6 theme plasmin-cleavage 1348:1363 arg1 fragments 1365:1373 the other plasmin-cleavage fragments 1338:1373 the other plasmin-cleavage fragments 1338:1373 As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments, did not retain the ability of intact C1-s to bind Ca2+. 2141278 3 7 theme growth 688:693 arg1 region 718:723 the epidermal growth factor-like (EGF-like) region 674:723 the epidermal growth factor-like (EGF-like) region of C1-s 674:731 Fragment alpha 2, which comprised the epidermal growth factor-like (EGF-like) region of C1-s, was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195. 2141278 0 8 theme factor 95:100 arg1 region 111:116 the epidermal growth factor homology region 74:116 the epidermal growth factor homology region 74:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 5 9 theme equilibrium 1295:1305 arg1 dialysis 1307:1314 equilibrium dialysis 1295:1314 equilibrium dialysis 1295:1314 As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments, did not retain the ability of intact C1-s to bind Ca2+. 2141278 1 10 link disulfide-linked 235:250 arg1 chains 252:257 two disulfide-linked chains 231:257 two disulfide-linked chains 231:257 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 10 link disulfide-linked 235:250 arg1 chain 314:318 the A chain 308:318 the A chain 308:318 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 10 link disulfide-linked 235:250 arg1 chain 266:270 the B chain 260:270 the B chain 260:270 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 4 11 theme amino 985:989 arg1 analysis 996:1003 amino acid analysis 985:1003 amino acid analysis 985:1003 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 0 12 theme growth 88:93 arg1 factor 95:100 epidermal growth factor 78:100 the epidermal growth factor homology region 74:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 1 13 theme disulfide-linked 235:250 arg1 chains 252:257 two disulfide-linked chains 231:257 two disulfide-linked chains 231:257 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 13 theme disulfide-linked 235:250 arg1 chain 314:318 the A chain 308:318 the A chain 308:318 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 13 theme disulfide-linked 235:250 arg1 chain 266:270 the B chain 260:270 the B chain 260:270 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 14 theme A 312:312 arg1 chains 252:257 two disulfide-linked chains 231:257 two disulfide-linked chains 231:257 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 14 theme A 312:312 arg1 chain 314:318 the A chain 308:318 the A chain 308:318 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 0 15 theme homology 102:109 arg1 region 111:116 the epidermal growth factor homology region 74:116 the epidermal growth factor homology region 74:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 7 16 theme alpha 1604:1608 arg1 fragment 1612:1619 the N-terminal alpha 1 fragment 1589:1619 the N-terminal alpha 1 fragment 1589:1619 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 16 theme alpha 1604:1608 arg1 able 1665:1668 able 1665:1668 able 1665:1668 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 17 theme A 1651:1651 arg1 chain 1653:1657 the A chain 1647:1657 the A chain 1647:1657 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 2 18 theme A 575:575 arg1 chain 577:581 the A chain 571:581 the A chain 571:581 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 4 19 with contrast 1176:1183 arg1 150 1215:1217 150 1215:1217 150 1215:1217 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 4 19 with contrast 1176:1183 arg1 position 1205:1212 the homologous position 1190:1212 the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine 1190:1277 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 1 20 theme C1- 159:161 arg1 subcomponents 142:154 the three subcomponents 132:154 the three subcomponents of C1-, the first component of complement 132:196 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 2 21 theme limited 514:520 arg1 proteolysis 522:532 limited proteolysis 514:532 limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma 514:637 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 5 22 theme native 1317:1322 arg1 alpha 1324:1328 native alpha 2 1317:1330 native alpha 2 1317:1330 As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments, did not retain the ability of intact C1-s to bind Ca2+. 2141278 0 23 theme Chemical 0:7 arg1 characterization 24:39 Chemical and functional characterization 0:39 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 0:117 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 6 24 theme C1-s 1491:1494 arg1 ability 1480:1486 the ability 1476:1486 the ability of C1-s to dimerize or to associate with C1-r in the presence of Ca2+ 1476:1556 In the same way, plasmin cleavage abolished the ability of C1-s to dimerize or to associate with C1-r in the presence of Ca2+. 2141278 1 25 contain containing 273:282 arg1 chains 252:257 two disulfide-linked chains 231:257 two disulfide-linked chains 231:257 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 25 contain containing 273:282 arg2 site 298:301 the catalytic site 284:301 the catalytic site 284:301 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 25 contain containing 273:282 arg1 chain 266:270 the B chain 260:270 the B chain 260:270 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 7 26 theme intact 1700:1705 arg1 C1s 1707:1709 intact C1s 1700:1709 intact C1s 1700:1709 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 6 27 theme Ca2+ 1553:1556 arg1 presence 1541:1548 the presence 1537:1548 the presence of Ca2+ 1537:1556 In the same way, plasmin cleavage abolished the ability of C1-s to dimerize or to associate with C1-r in the presence of Ca2+. 2141278 2 28 theme responsible 454:464 arg1 regions 446:452 the regions 442:452 the regions responsible for the latter functions 442:489 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 4 29 theme mass 1029:1032 arg1 spectrometry 1034:1045 mass spectrometry 1029:1045 mass spectrometry 1029:1045 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 1 30 with binding 338:344 arg1 subcomponents 401:413 the other C1- subcomponents 387:413 the other C1- subcomponents 387:413 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 31 theme first 168:172 arg1 component 174:182 the first component 164:182 the first component of complement 164:196 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 31 theme first 168:172 arg1 C1- 159:161 C1- 159:161 C1- 159:161 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 0 32 theme functional 13:22 arg1 characterization 24:39 Chemical and functional characterization 0:39 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 0:117 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 4 33 theme homologous 1194:1203 arg1 150 1215:1217 150 1215:1217 150 1215:1217 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 4 33 theme homologous 1194:1203 arg1 position 1205:1212 the homologous position 1190:1212 the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine 1190:1277 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 7 34 theme chain 1653:1657 arg1 serine 1634:1639 serine 24 1634:1642 serine 24 of the A chain 1634:1657 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 35 theme tetramer 1772:1779 arg1 formation 1719:1727 the formation 1715:1727 the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer 1715:1779 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 36 theme -dependent 1742:1751 arg1 tetramer 1772:1779 the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer 1732:1779 the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer 1732:1779 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 1 37 theme B 264:264 arg1 chains 252:257 two disulfide-linked chains 231:257 two disulfide-linked chains 231:257 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 37 theme B 264:264 arg1 chain 266:270 the B chain 260:270 the B chain 260:270 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 2 38 with proteolysis 522:532 arg1 plasmin 539:545 plasmin 539:545 plasmin 539:545 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 2 38 with proteolysis 522:532 arg1 treatment 550:558 a treatment 548:558 a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma 548:637 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 5 39 theme intact 1406:1411 arg1 C1-s 1413:1416 intact C1-s 1406:1416 intact C1-s 1406:1416 As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments, did not retain the ability of intact C1-s to bind Ca2+. 2141278 1 40 with interaction 367:377 arg1 subcomponents 401:413 the other C1- subcomponents 387:413 the other C1- subcomponents 387:413 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 5 41 theme other 1342:1346 arg1 fragments 1365:1373 the other plasmin-cleavage fragments 1338:1373 the other plasmin-cleavage fragments 1338:1373 As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments, did not retain the ability of intact C1-s to bind Ca2+. 2141278 1 42 theme Ca2+ 333:336 arg1 binding 338:344 Ca2+ binding 333:344 Ca2+ binding 333:344 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 43 theme other 391:395 arg1 subcomponents 401:413 the other C1- subcomponents 387:413 the other C1- subcomponents 387:413 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 44 theme complement 187:196 arg1 component 174:182 the first component 164:182 the first component of complement 164:196 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 44 theme complement 187:196 arg1 C1- 159:161 C1- 159:161 C1- 159:161 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 5 45 theme C1-s 1413:1416 arg1 ability 1395:1401 the ability 1391:1401 the ability of intact C1-s to bind Ca2+ 1391:1429 As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments, did not retain the ability of intact C1-s to bind Ca2+. 2141278 3 46 theme Fragment 640:647 arg1 alpha 649:653 Fragment alpha 2 640:655 Fragment alpha 2 640:655 Fragment alpha 2, which comprised the epidermal growth factor-like (EGF-like) region of C1-s, was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195. 2141278 6 47 theme plasmin 1449:1455 arg1 cleavage 1457:1464 plasmin cleavage 1449:1464 plasmin cleavage 1449:1464 In the same way, plasmin cleavage abolished the ability of C1-s to dimerize or to associate with C1-r in the presence of Ca2+. 2141278 1 48 theme C1- 397:399 arg1 subcomponents 401:413 the other C1- subcomponents 387:413 the other C1- subcomponents 387:413 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 0 49 theme fragment 46:53 arg1 characterization 24:39 Chemical and functional characterization 0:39 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 0:117 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 2 50 theme latter 474:479 arg1 functions 481:489 the latter functions 470:489 the latter functions 470:489 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 4 51 contain contains 1239:1246 arg1 C1-r 1223:1226 C1-r 1223:1226 C1-r which only contains erythro-beta-hydroxyasparagine 1223:1277 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 4 51 contain contains 1239:1246 arg2 erythro-beta-hydroxyasparagine 1248:1277 erythro-beta-hydroxyasparagine 1248:1277 erythro-beta-hydroxyasparagine 1248:1277 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 3 52 theme EGF-like 708:715 arg1 region 718:723 the epidermal growth factor-like (EGF-like) region 674:723 the epidermal growth factor-like (EGF-like) region of C1-s 674:731 Fragment alpha 2, which comprised the epidermal growth factor-like (EGF-like) region of C1-s, was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195. 2141278 7 53 from able 1665:1668 arg1 contrast 1562:1569 contrast 1562:1569 contrast 1562:1569 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 54 theme N-terminal 1593:1602 arg1 fragment 1612:1619 the N-terminal alpha 1 fragment 1589:1619 the N-terminal alpha 1 fragment 1589:1619 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 54 theme N-terminal 1593:1602 arg1 able 1665:1668 able 1665:1668 able 1665:1668 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 1 55 theme -dependent 356:365 arg1 interaction 367:377 Ca2(+)-dependent interaction 350:377 Ca2(+)-dependent interaction(s) with the other C1- subcomponents 350:413 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 55 theme -dependent 356:365 arg1 s 379:379 s 379:379 s 379:379 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 4 56 theme C1-r 1223:1226 arg1 150 1215:1217 150 1215:1217 150 1215:1217 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 4 56 theme C1-r 1223:1226 arg1 position 1205:1212 the homologous position 1190:1212 the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine 1190:1277 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 4 57 theme Edman 1006:1010 arg1 degradation 1012:1022 Edman degradation 1006:1022 Edman degradation 1006:1022 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 6 58 theme same 1439:1442 arg1 way 1444:1446 the same way 1435:1446 the same way 1435:1446 In the same way, plasmin cleavage abolished the ability of C1-s to dimerize or to associate with C1-r in the presence of Ca2+. 2141278 2 59 theme major 594:598 arg1 gamma 633:637 gamma 633:637 gamma 633:637 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 2 59 theme major 594:598 arg1 alpha 611:615 alpha 1 611:617 alpha 1 611:617 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 2 59 theme major 594:598 arg1 alpha 620:624 alpha 2 620:626 alpha 2 620:626 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 2 59 theme major 594:598 arg1 fragments 600:608 three major fragments 588:608 three major fragments 588:608 In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin, a treatment that split the A chain into three major fragments, alpha 1, alpha 2, and gamma. 2141278 3 60 theme factor-like 695:705 arg1 region 718:723 the epidermal growth factor-like (EGF-like) region 674:723 the epidermal growth factor-like (EGF-like) region of C1-s 674:731 Fragment alpha 2, which comprised the epidermal growth factor-like (EGF-like) region of C1-s, was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195. 2141278 4 61 theme C1-s 1077:1080 arg1 position 1061:1068 position 134 1061:1072 position 134 of C1-s 1061:1080 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 7 62 from contrast 1562:1569 arg1 fragment 1612:1619 the N-terminal alpha 1 fragment 1589:1619 the N-terminal alpha 1 fragment 1589:1619 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 7 62 from contrast 1562:1569 arg1 able 1665:1668 able 1665:1668 able 1665:1668 In contrast, both alpha 2 and the N-terminal alpha 1 fragment, starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2(+)-dependent C1-s-C1r-C1-r-C1-s tetramer. 2141278 0 63 theme C1-s 58:61 arg1 fragment 46:53 a fragment 44:53 a fragment of C1-s containing the epidermal growth factor homology region 44:116 Chemical and functional characterization of a fragment of C1-s containing the epidermal growth factor homology region. 2141278 1 64 theme catalytic 288:296 arg1 site 298:301 the catalytic site 284:301 the catalytic site 284:301 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 4 65 theme acid 991:994 arg1 analysis 996:1003 amino acid analysis 985:1003 amino acid analysis 985:1003 This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47%) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine. 2141278 3 66 theme C1-s 728:731 arg1 region 718:723 the epidermal growth factor-like (EGF-like) region 674:723 the epidermal growth factor-like (EGF-like) region of C1-s 674:731 Fragment alpha 2, which comprised the epidermal growth factor-like (EGF-like) region of C1-s, was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195. 2141278 1 67 theme serine 204:209 arg1 C1-s 119:122 C1-s 119:122 C1-s 119:122 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 2141278 1 67 theme serine 204:209 arg1 protease 211:218 a serine protease 202:218 a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents 202:413 C1-s, one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site, and the A chain, involved in Ca2+ binding and Ca2(+)-dependent interaction(s) with the other C1- subcomponents. 22848655 12 0 theme Met/HGF 1735:1741 arg1 complex 1743:1749 the Met/HGF complex 1731:1749 the Met/HGF complex 1731:1749 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 4 1 theme receptor 557:564 arg1 dimerization 566:577 receptor dimerization 557:577 receptor dimerization 557:577 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 7 2 theme Sema-PSI 1096:1103 arg1 domains 1105:1111 RON Sema-PSI domains 1092:1111 RON Sema-PSI domains 1092:1111 Here, we report the structure of RON Sema-PSI domains at 1.85 Å resolution. 22848655 11 3 theme crystal 1520:1526 arg1 packing 1528:1534 The RON Sema-PSI crystal packing 1503:1534 The RON Sema-PSI crystal packing 1503:1534 The RON Sema-PSI crystal packing generates a homodimer with interface formed by the Sema domain. 22848655 2 4 theme apoptosis 322:330 arg1 processes 332:340 cell adhesion, invasion, motility and apoptosis processes 284:340 cell adhesion, invasion, motility and apoptosis processes 284:340 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 11 5 theme RON 1507:1509 arg1 packing 1528:1534 The RON Sema-PSI crystal packing 1503:1534 The RON Sema-PSI crystal packing 1503:1534 The RON Sema-PSI crystal packing generates a homodimer with interface formed by the Sema domain. 22848655 8 6 theme Sema 1139:1142 arg1 domain 1144:1149 RON Sema domain 1135:1149 RON Sema domain 1135:1149 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 6 7 theme High 886:889 arg1 association 900:910 High affinity association 886:910 High affinity association between MSP and RON 886:930 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 8 8 theme disulfide 1203:1211 arg1 bond 1213:1216 disulfide bond rich 1203:1221 disulfide bond rich 1203:1221 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 8 8 theme disulfide 1203:1211 arg1 motif 1242:1246 cysteine-knot PSI motif 1224:1246 cysteine-knot PSI motif 1224:1246 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 2 9 theme signal 241:246 arg1 pathways 261:268 signal transduction pathways 241:268 signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes 241:340 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 8 10 theme rich 1218:1221 arg1 bond 1213:1216 disulfide bond rich 1203:1221 disulfide bond rich 1203:1221 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 8 10 theme rich 1218:1221 arg1 motif 1242:1246 cysteine-knot PSI motif 1224:1246 cysteine-knot PSI motif 1224:1246 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 2 11 theme adhesion 289:296 arg1 processes 332:340 cell adhesion, invasion, motility and apoptosis processes 284:340 cell adhesion, invasion, motility and apoptosis processes 284:340 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 1 12 theme cell 164:167 arg1 receptor 177:184 a cell surface receptor 162:184 a cell surface receptor for Macrophage Stimulating Protein (MSP) 162:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 12 theme cell 164:167 arg1 kinase 152:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 8 13 theme seven-bladed 1160:1171 arg1 fold 1185:1188 a seven-bladed β-propeller fold 1158:1188 a seven-bladed β-propeller fold 1158:1188 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 8 14 theme cysteine-knot 1224:1236 arg1 bond 1213:1216 disulfide bond rich 1203:1221 disulfide bond rich 1203:1221 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 8 14 theme cysteine-knot 1224:1236 arg1 motif 1242:1246 cysteine-knot PSI motif 1224:1246 cysteine-knot PSI motif 1224:1246 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 4 15 theme domains 645:651 arg1 autophosphorylation 597:615 autophosphorylation 597:615 autophosphorylation of the intracellular kinase domains 597:651 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 5 16 theme dimerization 715:726 arg1 domains 728:734 the ligand recognition and dimerization domains 688:734 domains 728:734 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 5 17 theme Immunoglobulins-Plexins-Transcription 826:862 arg1 domains 877:883 four Immunoglobulins-Plexins-Transcription factor (IPT) domains 821:883 four Immunoglobulins-Plexins-Transcription factor (IPT) domains 821:883 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 13 18 theme splice 2072:2077 arg1 RONΔ85 2088:2093 RONΔ85 2088:2093 RONΔ85 2088:2093 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 13 18 theme splice 2072:2077 arg1 variant 2079:2085 the soluble RON splice variant 2056:2085 the soluble RON splice variant 2056:2085 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 1 19 theme Recepteur 105:113 arg1 Nantais 125:131 Recepteur d'Origine Nantais 105:131 Recepteur d'Origine Nantais 105:131 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 19 theme Recepteur 105:113 arg1 RON 100:102 Human RON 94:102 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 20 theme Origine 117:123 arg1 Nantais 125:131 Recepteur d'Origine Nantais 105:131 Recepteur d'Origine Nantais 105:131 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 20 theme Origine 117:123 arg1 RON 100:102 Human RON 94:102 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 12 21 theme Growth 1689:1694 arg1 HGF 1704:1706 HGF 1704:1706 HGF 1704:1706 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 12 21 theme Growth 1689:1694 arg1 Factor 1696:1701 Hepatocyte Growth Factor 1678:1701 Hepatocyte Growth Factor (HGF) 1678:1707 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 6 22 theme RON 1006:1008 arg1 activation 1010:1019 RON activation 1006:1019 RON activation 1006:1019 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 0 23 theme human 58:62 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 11 24 with homodimer 1548:1556 arg1 interface 1563:1571 interface 1563:1571 interface formed by the Sema domain 1563:1597 The RON Sema-PSI crystal packing generates a homodimer with interface formed by the Sema domain. 22848655 0 25 theme receptor 68:75 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 13 26 theme soluble 2060:2066 arg1 RONΔ85 2088:2093 RONΔ85 2088:2093 RONΔ85 2088:2093 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 13 26 theme soluble 2060:2066 arg1 variant 2079:2085 the soluble RON splice variant 2056:2085 the soluble RON splice variant 2056:2085 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 1 27 theme tyrosine 143:150 arg1 receptor 177:184 a cell surface receptor 162:184 a cell surface receptor for Macrophage Stimulating Protein (MSP) 162:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 27 theme tyrosine 143:150 arg1 kinase 152:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 12 28 theme MSP 1662:1664 arg1 homology 1666:1673 MSP homology 1662:1673 MSP homology to Hepatocyte Growth Factor (HGF) 1662:1707 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 6 29 theme RON 987:989 arg1 Sema 991:994 RON Sema 987:994 RON Sema 987:994 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 4 30 theme MSP 485:487 arg1 heterodimer 493:503 MSP α/β heterodimer 485:503 MSP α/β heterodimer 485:503 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 13 31 theme ligand-independent 1937:1954 arg1 activation 1965:1974 ligand-independent receptor activation 1937:1974 ligand-independent receptor activation 1937:1974 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 9 32 with Comparison 1249:1258 arg1 kinase 1302:1307 the homologous Met receptor tyrosine kinase 1265:1307 the homologous Met receptor tyrosine kinase 1265:1307 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 0 33 theme kinase 86:91 arg1 domain 48:53 the Sema-PSI extracellular domain 21:53 the Sema-PSI extracellular domain of human RON receptor tyrosine kinase 21:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 0 33 theme kinase 86:91 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 9 34 theme homologous 1269:1278 arg1 receptor 1284:1291 the homologous Met receptor 1265:1291 the homologous Met receptor tyrosine kinase 1265:1307 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 9 35 theme receptor 1284:1291 arg1 kinase 1302:1307 the homologous Met receptor tyrosine kinase 1265:1307 the homologous Met receptor tyrosine kinase 1265:1307 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 3 36 theme RON 362:364 arg1 levels 352:357 Elevated levels 343:357 Elevated levels of RON and its alternatively spliced variants 343:403 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 0 37 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 0:92 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 6 38 theme MSP 1045:1047 arg1 proteins 1049:1056 MSP proteins 1045:1056 MSP proteins 1045:1056 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 13 39 theme receptor 1956:1963 arg1 activation 1965:1974 ligand-independent receptor activation 1937:1974 ligand-independent receptor activation 1937:1974 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 12 40 theme MSPβ 1807:1810 arg1 site 1820:1823 the putative MSPβ binding site 1794:1823 the putative MSPβ binding site 1794:1823 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 1 41 theme RON 100:102 arg1 receptor 177:184 a cell surface receptor 162:184 a cell surface receptor for Macrophage Stimulating Protein (MSP) 162:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 41 theme RON 100:102 arg1 kinase 152:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 12 42 theme interface 1621:1629 arg1 Mapping 1600:1606 Mapping 1600:1606 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex 1600:1749 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 3 43 theme spliced 388:394 arg1 variants 396:403 its alternatively spliced variants 370:403 its alternatively spliced variants 370:403 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 4 44 theme RON 536:538 arg1 receptor 540:547 RON receptor 536:547 RON receptor 536:547 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 7 45 theme Å 1121:1121 arg1 resolution 1123:1132 1.85 Å resolution 1116:1132 1.85 Å resolution 1116:1132 Here, we report the structure of RON Sema-PSI domains at 1.85 Å resolution. 22848655 0 46 theme Sema-PSI 25:32 arg1 domain 48:53 the Sema-PSI extracellular domain 21:53 the Sema-PSI extracellular domain of human RON receptor tyrosine kinase 21:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 0 46 theme Sema-PSI 25:32 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 5 47 contain containing 677:686 arg2 domains 728:734 the ligand recognition and dimerization domains 688:734 domains 728:734 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 5 47 contain containing 677:686 arg1 RON 672:674 RON 672:674 RON 672:674 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 5 47 contain containing 677:686 arg2 recognition 699:709 the ligand recognition and dimerization domains 688:734 recognition 699:709 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 5 48 theme Plexins-Semaphorins-Integrins 773:801 arg1 PSI 811:813 PSI 811:813 PSI 811:813 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 5 48 theme Plexins-Semaphorins-Integrins 773:801 arg1 domain 803:808 Plexins-Semaphorins-Integrins domain 773:808 Plexins-Semaphorins-Integrins domain (PSI) 773:814 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 9 49 theme distinguishing 1344:1357 arg1 features 1380:1387 distinguishing secondary structural features 1344:1387 distinguishing secondary structural features 1344:1387 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 13 50 theme activity 2018:2025 arg1 inhibition 1987:1996 the inhibition 1983:1996 the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85 1983:2093 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 3 51 theme tumor 457:461 arg1 cells 463:467 tumor cells 457:467 tumor cells 457:467 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 1 52 theme Stimulating 201:211 arg1 MSP 222:224 MSP 222:224 MSP 222:224 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 52 theme Stimulating 201:211 arg1 Protein 213:219 Macrophage Stimulating Protein 190:219 Macrophage Stimulating Protein (MSP) 190:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 13 53 theme variant 2045:2051 arg1 activity 2018:2025 the constitutive activity 2001:2025 the constitutive activity of RONΔ160 splice variant 2001:2051 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 12 54 theme complex 1743:1749 arg1 structure 1718:1726 the structure 1714:1726 the structure of the Met/HGF complex 1714:1749 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 12 54 theme complex 1743:1749 arg1 homology 1666:1673 MSP homology 1662:1673 MSP homology to Hepatocyte Growth Factor (HGF) 1662:1707 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 12 54 theme complex 1743:1749 arg1 homology 1645:1652 the RON homology 1637:1652 the RON homology to Met 1637:1659 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 4 55 theme heterodimer 493:503 arg1 binding 474:480 The binding 470:480 The binding of MSP α/β heterodimer to the extracellular region of RON receptor 470:547 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 7 56 theme RON 1092:1094 arg1 domains 1105:1111 RON Sema-PSI domains 1092:1111 RON Sema-PSI domains 1092:1111 Here, we report the structure of RON Sema-PSI domains at 1.85 Å resolution. 22848655 6 57 theme affinity 891:898 arg1 association 900:910 High affinity association 886:910 High affinity association between MSP and RON 886:930 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 11 58 theme Sema-PSI 1511:1518 arg1 packing 1528:1534 The RON Sema-PSI crystal packing 1503:1534 The RON Sema-PSI crystal packing 1503:1534 The RON Sema-PSI crystal packing generates a homodimer with interface formed by the Sema domain. 22848655 2 59 theme transduction 248:259 arg1 pathways 261:268 signal transduction pathways 241:268 signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes 241:340 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 5 60 theme factor 864:869 arg1 domains 877:883 four Immunoglobulins-Plexins-Transcription factor (IPT) domains 821:883 four Immunoglobulins-Plexins-Transcription factor (IPT) domains 821:883 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 8 61 theme RON 1135:1137 arg1 domain 1144:1149 RON Sema domain 1135:1149 RON Sema domain 1135:1149 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 13 62 theme dimer 1903:1907 arg1 assembly 1909:1916 the dimer assembly 1899:1916 the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85 1899:2093 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 5 63 theme IPT 872:874 arg1 domains 877:883 four Immunoglobulins-Plexins-Transcription factor (IPT) domains 821:883 four Immunoglobulins-Plexins-Transcription factor (IPT) domains 821:883 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 6 64 theme MSP 971:973 arg1 β-chain 975:981 MSP β-chain 971:981 MSP β-chain 971:981 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 0 65 theme domain 48:53 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 0:92 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 5 66 theme ligand 692:697 arg1 recognition 699:709 the ligand recognition and dimerization domains 688:734 recognition 699:709 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 2 67 theme cell 284:287 arg1 adhesion 289:296 cell adhesion 284:296 cell adhesion 284:296 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 1 68 theme surface 169:175 arg1 receptor 177:184 a cell surface receptor 162:184 a cell surface receptor for Macrophage Stimulating Protein (MSP) 162:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 68 theme surface 169:175 arg1 kinase 152:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 8 69 theme β-propeller 1173:1183 arg1 fold 1185:1188 a seven-bladed β-propeller fold 1158:1188 a seven-bladed β-propeller fold 1158:1188 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 4 70 theme kinase 638:643 arg1 domains 645:651 the intracellular kinase domains 620:651 the intracellular kinase domains 620:651 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 5 71 theme RON 672:674 arg1 RON 672:674 RON 672:674 RON 672:674 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 5 71 theme RON 672:674 arg1 ectodomain 658:667 The ectodomain 654:667 The ectodomain of RON, containing the ligand recognition and dimerization domains, 654:735 The ectodomain of RON, containing the ligand recognition and dimerization domains, is composed of a semaphorin (Sema), Plexins-Semaphorins-Integrins domain (PSI), and four Immunoglobulins-Plexins-Transcription factor (IPT) domains. 22848655 13 72 theme Sema-PSI 1866:1873 arg1 homodimer 1875:1883 The crystallographically determined RON Sema-PSI homodimer 1826:1883 The crystallographically determined RON Sema-PSI homodimer 1826:1883 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 1 73 theme d' 115:116 arg1 Nantais 125:131 Recepteur d'Origine Nantais 105:131 Recepteur d'Origine Nantais 105:131 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 73 theme d' 115:116 arg1 RON 100:102 Human RON 94:102 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 0 74 theme RON 64:66 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 13 75 theme RON 1862:1864 arg1 homodimer 1875:1883 The crystallographically determined RON Sema-PSI homodimer 1826:1883 The crystallographically determined RON Sema-PSI homodimer 1826:1883 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 4 76 theme intracellular 624:636 arg1 domains 645:651 the intracellular kinase domains 620:651 the intracellular kinase domains 620:651 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 12 77 theme Hepatocyte 1678:1687 arg1 HGF 1704:1706 HGF 1704:1706 HGF 1704:1706 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 12 77 theme Hepatocyte 1678:1687 arg1 Factor 1696:1701 Hepatocyte Growth Factor 1678:1701 Hepatocyte Growth Factor (HGF) 1678:1707 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 9 78 theme structural 1369:1378 arg1 features 1380:1387 distinguishing secondary structural features 1344:1387 distinguishing secondary structural features 1344:1387 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 0 79 theme tyrosine 77:84 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 1 80 theme receptor 134:141 arg1 receptor 177:184 a cell surface receptor 162:184 a cell surface receptor for Macrophage Stimulating Protein (MSP) 162:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 80 theme receptor 134:141 arg1 kinase 152:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 4 81 theme α/β 489:491 arg1 heterodimer 493:503 MSP α/β heterodimer 485:503 MSP α/β heterodimer 485:503 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 13 82 theme determined 1851:1860 arg1 homodimer 1875:1883 The crystallographically determined RON Sema-PSI homodimer 1826:1883 The crystallographically determined RON Sema-PSI homodimer 1826:1883 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 13 83 theme RON 2068:2070 arg1 RONΔ85 2088:2093 RONΔ85 2088:2093 RONΔ85 2088:2093 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 13 83 theme RON 2068:2070 arg1 variant 2079:2085 the soluble RON splice variant 2056:2085 the soluble RON splice variant 2056:2085 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 3 84 theme Elevated 343:350 arg1 levels 352:357 Elevated levels 343:357 Elevated levels of RON and its alternatively spliced variants 343:403 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 7 85 from resolution 1123:1132 arg1 structure 1079:1087 the structure 1075:1087 the structure of RON Sema-PSI domains at 1.85 Å resolution 1075:1132 Here, we report the structure of RON Sema-PSI domains at 1.85 Å resolution. 22848655 10 86 theme exclusive 1418:1426 arg1 selectivity 1428:1438 the receptors' exclusive selectivity 1403:1438 the receptors' exclusive selectivity towards their respective ligands, RON for MSP and Met for HGF 1403:1500 These define the receptors' exclusive selectivity towards their respective ligands, RON for MSP and Met for HGF. 22848655 3 87 theme cells 463:467 arg1 metastasis 443:452 metastasis 443:452 metastasis 443:452 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 3 87 theme cells 463:467 arg1 progression 427:437 progression 427:437 progression 427:437 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 9 88 theme Met 1280:1282 arg1 receptor 1284:1291 the homologous Met receptor 1265:1291 the homologous Met receptor tyrosine kinase 1265:1307 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 9 89 theme tyrosine 1293:1300 arg1 kinase 1302:1307 the homologous Met receptor tyrosine kinase 1265:1307 the homologous Met receptor tyrosine kinase 1265:1307 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 12 90 theme RON 1641:1643 arg1 homology 1645:1652 the RON homology 1637:1652 the RON homology to Met 1637:1659 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 10 91 theme respective 1454:1463 arg1 ligands 1465:1471 their respective ligands 1448:1471 their respective ligands 1448:1471 These define the receptors' exclusive selectivity towards their respective ligands, RON for MSP and Met for HGF. 22848655 10 91 theme respective 1454:1463 arg1 RON 1474:1476 RON 1474:1476 RON for MSP and Met 1474:1492 These define the receptors' exclusive selectivity towards their respective ligands, RON for MSP and Met for HGF. 22848655 13 92 theme RONΔ160 2030:2036 arg1 variant 2045:2051 RONΔ160 splice variant 2030:2051 RONΔ160 splice variant 2030:2051 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 12 93 theme binding 1812:1818 arg1 site 1820:1823 the putative MSPβ binding site 1794:1823 the putative MSPβ binding site 1794:1823 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 1 94 theme Human 94:98 arg1 Nantais 125:131 Recepteur d'Origine Nantais 105:131 Recepteur d'Origine Nantais 105:131 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 94 theme Human 94:98 arg1 RON 100:102 Human RON 94:102 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase 94:157 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 4 95 theme receptor 540:547 arg1 region 526:531 the extracellular region 508:531 the extracellular region of RON receptor 508:547 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 0 96 theme extracellular 34:46 arg1 domain 48:53 the Sema-PSI extracellular domain 21:53 the Sema-PSI extracellular domain of human RON receptor tyrosine kinase 21:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 0 96 theme extracellular 34:46 arg1 kinase 86:91 human RON receptor tyrosine kinase 58:91 human RON receptor tyrosine kinase 58:91 Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. 22848655 9 97 theme RON 1322:1324 arg1 Sema-PSI 1326:1333 RON Sema-PSI 1322:1333 RON Sema-PSI 1322:1333 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 12 98 theme dimer 1615:1619 arg1 interface 1621:1629 the dimer interface 1611:1629 the dimer interface 1611:1629 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 6 99 theme intact 1030:1035 arg1 RON 1037:1039 intact RON 1030:1039 intact RON 1030:1039 High affinity association between MSP and RON is mediated by the interaction between MSP β-chain and RON Sema, although RON activation requires intact RON and MSP proteins. 22848655 3 100 theme variants 396:403 arg1 levels 352:357 Elevated levels 343:357 Elevated levels of RON and its alternatively spliced variants 343:403 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 7 101 theme domains 1105:1111 arg1 structure 1079:1087 the structure 1075:1087 the structure of RON Sema-PSI domains at 1.85 Å resolution 1075:1132 Here, we report the structure of RON Sema-PSI domains at 1.85 Å resolution. 22848655 13 102 theme splice 2038:2043 arg1 variant 2045:2051 RONΔ160 splice variant 2030:2051 RONΔ160 splice variant 2030:2051 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 4 103 theme extracellular 512:524 arg1 region 526:531 the extracellular region 508:531 the extracellular region of RON receptor 508:547 The binding of MSP α/β heterodimer to the extracellular region of RON receptor induces receptor dimerization and activation by autophosphorylation of the intracellular kinase domains. 22848655 13 104 theme constitutive 2005:2016 arg1 activity 2018:2025 the constitutive activity 2001:2025 the constitutive activity of RONΔ160 splice variant 2001:2051 The crystallographically determined RON Sema-PSI homodimer may represent the dimer assembly that occurs during ligand-independent receptor activation and/or the inhibition of the constitutive activity of RONΔ160 splice variant by the soluble RON splice variant, RONΔ85. 22848655 2 105 theme motility 309:316 arg1 processes 332:340 cell adhesion, invasion, motility and apoptosis processes 284:340 cell adhesion, invasion, motility and apoptosis processes 284:340 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 9 106 theme secondary 1359:1367 arg1 features 1380:1387 distinguishing secondary structural features 1344:1387 distinguishing secondary structural features 1344:1387 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 12 107 theme putative 1798:1805 arg1 site 1820:1823 the putative MSPβ binding site 1794:1823 the putative MSPβ binding site 1794:1823 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 3 108 dep progression 427:437 arg1 the 423:425 the 423:425 the 423:425 Elevated levels of RON and its alternatively spliced variants are implicated in the progression and metastasis of tumor cells. 22848655 12 109 theme dimer 1761:1765 arg1 interface 1767:1775 the dimer interface 1757:1775 the dimer interface overlapping with the putative MSPβ binding site 1757:1823 Mapping of the dimer interface using the RON homology to Met, MSP homology to Hepatocyte Growth Factor (HGF), and the structure of the Met/HGF complex shows the dimer interface overlapping with the putative MSPβ binding site. 22848655 9 110 contain contains 1335:1342 arg2 features 1380:1387 distinguishing secondary structural features 1344:1387 distinguishing secondary structural features 1344:1387 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 9 110 contain contains 1335:1342 arg1 Sema-PSI 1326:1333 RON Sema-PSI 1322:1333 RON Sema-PSI 1322:1333 Comparison with the homologous Met receptor tyrosine kinase reveals that RON Sema-PSI contains distinguishing secondary structural features. 22848655 8 111 theme PSI 1238:1240 arg1 bond 1213:1216 disulfide bond rich 1203:1221 disulfide bond rich 1203:1221 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 8 111 theme PSI 1238:1240 arg1 motif 1242:1246 cysteine-knot PSI motif 1224:1246 cysteine-knot PSI motif 1224:1246 RON Sema domain adopts a seven-bladed β-propeller fold, followed by disulfide bond rich, cysteine-knot PSI motif. 22848655 2 112 theme invasion 299:306 arg1 processes 332:340 cell adhesion, invasion, motility and apoptosis processes 284:340 cell adhesion, invasion, motility and apoptosis processes 284:340 RON mediates signal transduction pathways that regulate cell adhesion, invasion, motility and apoptosis processes. 22848655 1 113 theme Macrophage 190:199 arg1 MSP 222:224 MSP 222:224 MSP 222:224 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 1 113 theme Macrophage 190:199 arg1 Protein 213:219 Macrophage Stimulating Protein 190:219 Macrophage Stimulating Protein (MSP) 190:225 Human RON (Recepteur d'Origine Nantais) receptor tyrosine kinase is a cell surface receptor for Macrophage Stimulating Protein (MSP). 22848655 11 114 theme Sema 1587:1590 arg1 domain 1592:1597 the Sema domain 1583:1597 the Sema domain 1583:1597 The RON Sema-PSI crystal packing generates a homodimer with interface formed by the Sema domain. 28661582 0 0 theme Active 73:78 arg1 Site 80:83 a Binuclear Zinc Active Site 56:83 a Binuclear Zinc Active Site Important for Melanogenesis 56:111 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 1 1 theme melanin 251:257 arg1 production 237:246 the production 233:246 the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair 233:327 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 4 2 theme factor-like 896:906 arg1 fold 908:911 an epidermal growth factor-like fold 876:911 an epidermal growth factor-like fold 876:911 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 1 3 theme hair 324:327 arg1 pigmentation 293:304 the pigmentation 289:304 the pigmentation of skin, eye, and hair 289:327 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 0 4 theme Important 85:93 arg1 Site 80:83 a Binuclear Zinc Active Site 56:83 a Binuclear Zinc Active Site Important for Melanogenesis 56:111 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 3 5 contain contains 578:585 arg2 ions 596:599 two zinc ions 587:599 two zinc ions in the active site instead of copper ions 587:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 5 contain contains 578:585 arg1 TYRP1 572:576 TYRP1 572:576 TYRP1 572:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 5 contain contains 578:585 arg1 subdomain 559:567 the typical tyrosinase-like subdomain 531:567 the typical tyrosinase-like subdomain of TYRP1 531:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 2 6 theme pigmentation 487:498 arg1 disorders 500:508 pigmentation disorders 487:508 pigmentation disorders 487:508 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 3 7 from ions 596:599 arg1 ions 638:641 copper ions 631:641 copper ions 631:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 7 from ions 596:599 arg1 site 615:618 the active site 604:618 the active site instead of copper ions 604:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 2 8 theme pure 375:378 arg1 form 380:383 pure form 375:383 pure form 375:383 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 4 9 theme tyrosinase 948:957 arg1 subdomain 959:967 the tyrosinase subdomain 944:967 the tyrosinase subdomain 944:967 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 1 10 theme tyrosinase-like 167:181 arg1 glycoenzymes 183:194 three tyrosinase-like glycoenzymes 161:194 three tyrosinase-like glycoenzymes in human melanocytes 161:215 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 2 11 theme activity 426:433 arg1 understanding 403:415 the understanding 399:415 the understanding of their activity 399:433 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 2 11 theme activity 426:433 arg1 effect 443:448 the effect 439:448 the effect of mutations that cause albinism and pigmentation disorders 439:508 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 2 12 theme mutations 453:461 arg1 understanding 403:415 the understanding 399:415 the understanding of their activity 399:433 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 2 12 theme mutations 453:461 arg1 effect 443:448 the effect 439:448 the effect of mutations that cause albinism and pigmentation disorders 439:508 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 3 13 theme tyrosinase-like 543:557 arg1 TYRP1 572:576 TYRP1 572:576 TYRP1 572:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 13 theme tyrosinase-like 543:557 arg1 subdomain 559:567 the typical tyrosinase-like subdomain 531:567 the typical tyrosinase-like subdomain of TYRP1 531:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 1 14 theme glycoenzymes 183:194 arg1 glycoenzymes 183:194 three tyrosinase-like glycoenzymes 161:194 three tyrosinase-like glycoenzymes in human melanocytes 161:215 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 1 14 theme glycoenzymes 183:194 arg1 one 154:156 one 154:156 one 154:156 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 3 15 theme tyrosinase 710:719 arg1 activity 727:734 tyrosinase redox activity 710:734 tyrosinase redox activity 710:734 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 4 16 theme Cys-rich 800:807 arg1 subdomain 809:817 the Cys-rich subdomain 796:817 the Cys-rich subdomain 796:817 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 4 16 theme Cys-rich 800:807 arg1 unique 829:834 unique 829:834 unique 829:834 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 0 17 theme Tyrosinase 19:28 arg1 Protein 1 38:46 Human Tyrosinase Related Protein 1 13:46 Human Tyrosinase Related Protein 1 13:46 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 1 18 theme responsible 273:283 arg1 melanin 251:257 melanin 251:257 melanin 251:257 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 1 18 theme responsible 273:283 arg1 compound 264:271 the compound 260:271 the compound responsible for the pigmentation of skin, eye, and hair 260:327 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 4 19 theme first 780:784 arg1 time 786:789 the first time 776:789 the first time 776:789 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 1 20 theme human 199:203 arg1 melanocytes 205:215 human melanocytes 199:215 human melanocytes 199:215 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 0 21 theme Human 13:17 arg1 Protein 1 38:46 Human Tyrosinase Related Protein 1 13:46 Human Tyrosinase Related Protein 1 13:46 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 5 22 theme TYRP1 1033:1037 arg1 mutations 1020:1028 most albinism-related mutations 998:1028 most albinism-related mutations of TYRP1 998:1037 Our structures suggest that most albinism-related mutations of TYRP1 affect its stability or activity. 28661582 3 23 theme typical 535:541 arg1 TYRP1 572:576 TYRP1 572:576 TYRP1 572:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 23 theme typical 535:541 arg1 subdomain 559:567 the typical tyrosinase-like subdomain 531:567 the typical tyrosinase-like subdomain of TYRP1 531:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 0 24 theme Protein 1 38:46 arg1 Structure 0:8 Structure 0:8 Structure of Human Tyrosinase Related Protein 1 0:46 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 4 25 theme epidermal 879:887 arg1 fold 908:911 an epidermal growth factor-like fold 876:911 an epidermal growth factor-like fold 876:911 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 4 26 contain has 872:874 arg1 subdomain 809:817 the Cys-rich subdomain 796:817 the Cys-rich subdomain 796:817 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 4 26 contain has 872:874 arg1 unique 829:834 unique 829:834 unique 829:834 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 4 26 contain has 872:874 arg2 fold 908:911 an epidermal growth factor-like fold 876:911 an epidermal growth factor-like fold 876:911 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 0 27 theme Related 30:36 arg1 Protein 1 38:46 Human Tyrosinase Related Protein 1 13:46 Human Tyrosinase Related Protein 1 13:46 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 3 28 theme active 608:613 arg1 ions 638:641 copper ions 631:641 copper ions 631:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 28 theme active 608:613 arg1 site 615:618 the active site 604:618 the active site instead of copper ions 604:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 2 29 from enzymes 364:370 arg1 form 380:383 pure form 375:383 pure form 375:383 Difficulties with producing these enzymes in pure form have hampered the understanding of their activity and the effect of mutations that cause albinism and pigmentation disorders. 28661582 4 30 theme vertebrate 839:848 arg1 proteins 862:869 vertebrate melanogenic proteins 839:869 vertebrate melanogenic proteins 839:869 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 3 31 theme redox 721:725 arg1 activity 727:734 tyrosinase redox activity 710:734 tyrosinase redox activity 710:734 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 1 32 theme Tyrosinase-related 114:131 arg1 TYRP1 144:148 TYRP1 144:148 TYRP1 144:148 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 1 32 theme Tyrosinase-related 114:131 arg1 protein 1 133:141 Tyrosinase-related protein 1 114:141 Tyrosinase-related protein 1 (TYRP1) 114:149 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 1 33 theme skin 309:312 arg1 pigmentation 293:304 the pigmentation 289:304 the pigmentation of skin, eye, and hair 289:327 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 5 34 theme most 998:1001 arg1 mutations 1020:1028 most albinism-related mutations 998:1028 most albinism-related mutations of TYRP1 998:1037 Our structures suggest that most albinism-related mutations of TYRP1 affect its stability or activity. 28661582 3 35 theme TYRP1 572:576 arg1 TYRP1 572:576 TYRP1 572:576 TYRP1 572:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 35 theme TYRP1 572:576 arg1 subdomain 559:567 the typical tyrosinase-like subdomain 531:567 the typical tyrosinase-like subdomain of TYRP1 531:576 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 5 36 theme albinism-related 1003:1018 arg1 mutations 1020:1028 most albinism-related mutations 998:1028 most albinism-related mutations of TYRP1 998:1037 Our structures suggest that most albinism-related mutations of TYRP1 affect its stability or activity. 28661582 4 37 theme melanogenic 850:860 arg1 proteins 862:869 vertebrate melanogenic proteins 839:869 vertebrate melanogenic proteins 839:869 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 28661582 3 38 theme copper 631:636 arg1 ions 638:641 copper ions 631:641 copper ions 631:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 1 39 from glycoenzymes 183:194 arg1 melanocytes 205:215 human melanocytes 199:215 human melanocytes 199:215 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 0 40 theme Zinc 68:71 arg1 Site 80:83 a Binuclear Zinc Active Site 56:83 a Binuclear Zinc Active Site Important for Melanogenesis 56:111 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 3 41 theme ions 638:641 arg1 ions 638:641 copper ions 631:641 copper ions 631:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 41 theme ions 638:641 arg1 site 615:618 the active site 604:618 the active site instead of copper ions 604:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 3 42 theme zinc 591:594 arg1 ions 596:599 two zinc ions 587:599 two zinc ions in the active site instead of copper ions 587:641 Herein we show that the typical tyrosinase-like subdomain of TYRP1 contains two zinc ions in the active site instead of copper ions as found in tyrosinases, which explains why TYRP1 does not exhibit tyrosinase redox activity. 28661582 0 43 theme Binuclear 58:66 arg1 Site 80:83 a Binuclear Zinc Active Site 56:83 a Binuclear Zinc Active Site Important for Melanogenesis 56:111 Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis. 28661582 1 44 theme eye 315:317 arg1 pigmentation 293:304 the pigmentation 289:304 the pigmentation of skin, eye, and hair 289:327 Tyrosinase-related protein 1 (TYRP1) is one of three tyrosinase-like glycoenzymes in human melanocytes that are key to the production of melanin, the compound responsible for the pigmentation of skin, eye, and hair. 28661582 4 45 theme growth 889:894 arg1 fold 908:911 an epidermal growth factor-like fold 876:911 an epidermal growth factor-like fold 876:911 In addition, the structures reveal for the first time that the Cys-rich subdomain, which is unique to vertebrate melanogenic proteins, has an epidermal growth factor-like fold and is tightly associated with the tyrosinase subdomain. 32815518 0 0 theme unique 70:75 arg1 mechanism 86:94 a unique assembly mechanism 68:94 a unique assembly mechanism 68:94 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 3 1 theme Å 655:655 arg1 rise 643:646 a rise 641:646 a rise of ~65 Å 641:655 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 1 theme Å 655:655 arg1 twist 663:667 a twist 661:667 a twist of ~180° 661:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 4 2 theme preceding 797:805 arg1 ZPC 807:809 the preceding ZPC 793:809 the preceding ZPC 793:809 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 1 3 from protein 153:159 arg1 urine 170:174 human urine 164:174 human urine 164:174 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 2 4 theme immune 345:350 arg1 response 352:359 the innate immune response 334:359 the innate immune response against urinary tract infections 334:392 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 1 5 theme aggregate 232:240 arg1 uropathogens 242:253 and aggregate uropathogens 228:253 uropathogens 242:253 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 4 6 theme β-sheet 843:849 arg1 complementation 851:865 β-sheet complementation 843:865 β-sheet complementation 843:865 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 2 7 from role 326:329 arg1 response 352:359 the innate immune response 334:359 the innate immune response against urinary tract infections 334:392 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 2 8 theme innate 338:343 arg1 response 352:359 the innate immune response 334:359 the innate immune response against urinary tract infections 334:392 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 3 9 from structure 500:508 arg1 resolution 545:554 3.5 Å resolution 539:554 3.5 Å resolution 539:554 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 10 theme core 531:534 arg1 structure 500:508 the cryo-EM structure 488:508 the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180° 488:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 0 11 theme assembly 77:84 arg1 mechanism 86:94 a unique assembly mechanism 68:94 a unique assembly mechanism 68:94 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 5 12 theme UMOD 1075:1078 arg1 precursors 1080:1089 assembly-incompetent UMOD precursors 1054:1089 assembly-incompetent UMOD precursors 1054:1089 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 4 13 theme ZPN 703:705 arg1 monomer 734:740 each monomer 729:740 each monomer 729:740 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 4 13 theme ZPN 703:705 arg1 subdomains 715:724 The immunoglobulin-like ZPN and ZPC subdomains 679:724 The immunoglobulin-like ZPN and ZPC subdomains of each monomer 679:740 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 3 14 theme cryo-EM 492:498 arg1 structure 500:508 the cryo-EM structure 488:508 the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180° 488:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 5 15 from synchronized 972:983 arg1 mechanism 922:930 an assembly mechanism 910:930 an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane 910:1105 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 3 16 theme UMOD 517:520 arg1 core 531:534 the UMOD filament core 513:534 the UMOD filament core 513:534 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 17 with arrangement 624:634 arg1 rise 643:646 a rise 641:646 a rise of ~65 Å 641:655 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 17 with arrangement 624:634 arg1 twist 663:667 a twist 661:667 a twist of ~180° 661:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 1 18 theme abundant 144:151 arg1 protein 153:159 the most abundant protein 135:159 the most abundant protein in human urine 135:174 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 1 18 theme abundant 144:151 arg1 uromodulin 114:123 The glycoprotein uromodulin 97:123 The glycoprotein uromodulin (UMOD) 97:130 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 5 19 with synchronized 972:983 arg1 cleavage 1002:1009 proteolytic cleavage 990:1009 proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane 990:1105 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 3 20 theme filament 522:529 arg1 core 531:534 the UMOD filament core 513:534 the UMOD filament core 513:534 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 0 21 theme cryo-EM 4:10 arg1 structure 12:20 The cryo-EM structure 0:20 The cryo-EM structure of the human uromodulin filament core 0:58 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 5 22 theme unique 872:877 arg1 architecture 888:899 The unique filament architecture 868:899 The unique filament architecture 868:899 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 2 23 theme tract 377:381 arg1 infections 383:392 urinary tract infections 369:392 urinary tract infections 369:392 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 2 24 theme UMOD 433:436 arg1 polymerization 438:451 UMOD polymerization 433:451 UMOD polymerization 433:451 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 1 25 theme pathogen 266:273 arg1 clearance 275:283 pathogen clearance 266:283 pathogen clearance 266:283 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 5 26 theme filament 879:886 arg1 architecture 888:899 The unique filament architecture 868:899 The unique filament architecture 868:899 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 1 27 theme human 164:168 arg1 urine 170:174 human urine 164:174 human urine 164:174 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 5 28 theme proteolytic 990:1000 arg1 cleavage 1002:1009 proteolytic cleavage 990:1009 proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane 990:1105 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 4 29 theme immunoglobulin-like 683:701 arg1 ZPN 703:705 immunoglobulin-like ZPN 683:705 immunoglobulin-like ZPN 683:705 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 3 30 theme ~180° 672:676 arg1 rise 643:646 a rise 641:646 a rise of ~65 Å 641:655 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 30 theme ~180° 672:676 arg1 twist 663:667 a twist 661:667 a twist of ~180° 661:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 1 31 gly glycoprotein 101:112 arg1 glycoprotein 101:112 The glycoprotein uromodulin 97:123 The glycoprotein uromodulin (UMOD) 97:130 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 5 32 theme pro-peptide 1029:1039 arg1 cleavage 1002:1009 proteolytic cleavage 990:1009 proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane 990:1105 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 3 33 theme Å 543:543 arg1 resolution 545:554 3.5 Å resolution 539:554 3.5 Å resolution 539:554 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 5 34 theme assembly 913:920 arg1 mechanism 922:930 an assembly mechanism 910:930 an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane 910:1105 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 0 35 theme human 29:33 arg1 core 55:58 the human uromodulin filament core 25:58 the human uromodulin filament core 25:58 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 1 36 theme urine 288:292 arg1 excretion 294:302 urine excretion 288:302 urine excretion 288:302 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 2 37 theme urinary 369:375 arg1 infections 383:392 urinary tract infections 369:392 urinary tract infections 369:392 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 2 38 theme structural 399:408 arg1 basis 410:414 structural basis 399:414 structural basis 399:414 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 3 39 theme bipartite 574:582 arg1 pellucida 589:597 the bipartite zona pellucida 570:597 the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180° 570:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 39 theme bipartite 574:582 arg1 ZP 600:601 ZP 600:601 ZP 600:601 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 1 40 theme filamentous 186:196 arg1 homopolymers 198:209 filamentous homopolymers 186:209 filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion 186:302 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 0 41 theme filament 46:53 arg1 core 55:58 the human uromodulin filament core 25:58 the human uromodulin filament core 25:58 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 4 42 theme ZPN 825:827 arg1 subdomains 829:838 ZPN subdomains 825:838 ZPN subdomains 825:838 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 5 43 theme assembly-incompetent 1054:1073 arg1 precursors 1080:1089 assembly-incompetent UMOD precursors 1054:1089 assembly-incompetent UMOD precursors 1054:1089 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 4 44 theme long 761:764 arg1 linker 766:771 a long linker 759:771 a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation 759:865 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 1 45 theme glycoprotein 101:112 arg1 UMOD 126:129 UMOD 126:129 UMOD 126:129 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 1 45 theme glycoprotein 101:112 arg1 uromodulin 114:123 The glycoprotein uromodulin 97:123 The glycoprotein uromodulin (UMOD) 97:130 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 1 45 theme glycoprotein 101:112 arg1 protein 153:159 the most abundant protein 135:159 the most abundant protein in human urine 135:174 The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. 32815518 0 46 theme uromodulin 35:44 arg1 core 55:58 the human uromodulin filament core 25:58 the human uromodulin filament core 25:58 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 2 47 dep basis 410:414 arg1 the 395:397 the 395:397 the 395:397 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 4 48 theme monomer 734:740 arg1 monomer 734:740 each monomer 729:740 each monomer 729:740 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 4 48 theme monomer 734:740 arg1 subdomains 715:724 The immunoglobulin-like ZPN and ZPC subdomains 679:724 The immunoglobulin-like ZPN and ZPC subdomains of each monomer 679:740 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 4 49 theme ZPC 711:713 arg1 monomer 734:740 each monomer 729:740 each monomer 729:740 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 4 49 theme ZPC 711:713 arg1 subdomains 715:724 The immunoglobulin-like ZPN and ZPC subdomains 679:724 The immunoglobulin-like ZPN and ZPC subdomains of each monomer 679:740 The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. 32815518 3 50 theme helical 616:622 arg1 arrangement 624:634 a helical arrangement 614:634 a helical arrangement with a rise of ~65 Å and a twist of ~180° 614:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 5 51 theme subunit 941:947 arg1 incorporation 949:961 subunit incorporation 941:961 subunit incorporation 941:961 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 2 52 theme critical 317:324 arg1 role 326:329 its critical role 313:329 its critical role in the innate immune response against urinary tract infections 313:392 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 3 53 from resolution 545:554 arg1 core 531:534 the UMOD filament core 513:534 the UMOD filament core 513:534 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 53 from resolution 545:554 arg1 structure 500:508 the cryo-EM structure 488:508 the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180° 488:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 5 54 theme C-terminal 1018:1027 arg1 pro-peptide 1029:1039 the C-terminal pro-peptide 1014:1039 the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane 1014:1105 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 3 55 theme zona 584:587 arg1 pellucida 589:597 the bipartite zona pellucida 570:597 the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180° 570:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 3 55 theme zona 584:587 arg1 ZP 600:601 ZP 600:601 ZP 600:601 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 0 56 theme core 55:58 arg1 structure 12:20 The cryo-EM structure 0:20 The cryo-EM structure of the human uromodulin filament core 0:58 The cryo-EM structure of the human uromodulin filament core reveals a unique assembly mechanism. 32815518 3 57 theme pellucida 589:597 arg1 module 604:609 the bipartite zona pellucida (ZP) module 570:609 the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180° 570:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 32815518 2 58 theme polymerization 438:451 arg1 mechanism 420:428 mechanism 420:428 mechanism 420:428 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 2 58 theme polymerization 438:451 arg1 basis 410:414 structural basis 399:414 structural basis 399:414 Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. 32815518 5 59 from mechanism 922:930 arg1 synchronized 972:983 synchronized 972:983 synchronized 972:983 The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. 32815518 3 60 from module 604:609 arg1 arrangement 624:634 a helical arrangement 614:634 a helical arrangement with a rise of ~65 Å and a twist of ~180° 614:676 Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. 17650508 0 0 theme calpain 93:99 arg1 Characterization 0:15 Characterization 0:15 Characterization of the intracellular proteolytic cleavage of myocilin 0:69 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 0 0 theme calpain 93:99 arg1 identification 75:88 identification 75:88 identification of calpain II as a myocilin-processing protease 75:136 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 10 1 with fractionation 1322:1334 arg1 K 1392:1392 proteinase K 1381:1392 proteinase K 1381:1392 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 7 2 theme Intracellular 996:1008 arg1 uptake 1036:1041 Intracellular ionomycin-induced calcium uptake 996:1041 Intracellular ionomycin-induced calcium uptake 996:1041 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 11 3 from processing 1595:1604 arg1 lumen 1625:1629 the lumen 1621:1629 the lumen of the endoplasmic reticulum 1621:1658 These data revealed that calpain II is responsible for the intracellular processing of myocilin in the lumen of the endoplasmic reticulum. 17650508 1 4 theme unknown 238:244 arg1 function 246:253 unknown function 238:253 unknown function 238:253 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 3 5 from cleavage 509:516 arg1 region 532:537 the linker region 521:537 the linker region 521:537 We show herein that the proteolytic cleavage in the linker region splits the two terminal domains. 17650508 9 6 from cells 1217:1221 arg1 culture 1226:1232 culture 1226:1232 culture 1226:1232 However, in cells in culture, only RNA interference knockdown of calpain II reduced myocilin processing. 17650508 1 7 theme function 246:253 arg1 glycoprotein 222:233 a secreted glycoprotein 211:233 a secreted glycoprotein of unknown function 211:253 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 1 7 theme function 246:253 arg1 myocilin 201:208 myocilin 201:208 myocilin 201:208 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 10 8 theme reticulum 1467:1475 arg1 lumen 1442:1446 the lumen 1438:1446 the lumen of the endoplasmic reticulum together with a subpopulation of calpain II 1438:1519 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 10 9 with digestion 1340:1348 arg1 K 1392:1392 proteinase K 1381:1392 proteinase K 1381:1392 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 6 10 dep intra- 929:934 arg1 calcium 953:959 calcium 953:959 calcium 953:959 Since calpains are calcium-activated proteases, we analyzed how changes in either intra- or extracellular calcium affected the cleavage of myocilin. 17650508 0 11 theme myocilin-processing 109:127 arg1 protease 129:136 a myocilin-processing protease 107:136 a myocilin-processing protease 107:136 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 1 12 theme C-terminal 343:352 arg1 domain 372:377 a C-terminal olfactomedin-like domain 341:377 a C-terminal olfactomedin-like domain 341:377 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 7 13 theme proteolytic 1136:1146 arg1 processing 1148:1157 the proteolytic processing 1132:1157 the proteolytic processing 1132:1157 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 1 14 gly glycoprotein 222:233 arg1 glycoprotein 222:233 a secreted glycoprotein 211:233 a secreted glycoprotein of unknown function 211:253 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 1 14 gly glycoprotein 222:233 arg1 myocilin 201:208 myocilin 201:208 myocilin 201:208 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 12 15 theme extracellular 1706:1718 arg1 interactions 1720:1731 extracellular interactions 1706:1731 extracellular interactions of myocilin 1706:1743 We propose that this cleavage might regulate extracellular interactions of myocilin, contributing to the control of intraocular pressure. 17650508 1 16 theme different 164:172 arg1 types 174:178 different types 164:178 different types of glaucoma 164:190 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 1 17 theme olfactomedin-like 354:370 arg1 domain 372:377 a C-terminal olfactomedin-like domain 341:377 a C-terminal olfactomedin-like domain 341:377 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 5 18 theme calpain 808:814 arg1 inhibitor 816:824 calpain inhibitor IV 808:827 calpain inhibitor IV 808:827 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 12 19 theme myocilin 1736:1743 arg1 interactions 1720:1731 extracellular interactions 1706:1731 extracellular interactions of myocilin 1706:1743 We propose that this cleavage might regulate extracellular interactions of myocilin, contributing to the control of intraocular pressure. 17650508 2 20 theme endoproteolytic 445:459 arg1 processing 461:470 an intracellular endoproteolytic processing 428:470 an intracellular endoproteolytic processing 428:470 Recently, we have shown that myocilin undergoes an intracellular endoproteolytic processing. 17650508 9 21 theme only 1235:1238 arg1 knockdown 1257:1265 only RNA interference knockdown 1235:1265 only RNA interference knockdown of calpain II 1235:1279 However, in cells in culture, only RNA interference knockdown of calpain II reduced myocilin processing. 17650508 10 22 theme Subcellular 1310:1320 arg1 fractionation 1322:1334 Subcellular fractionation 1310:1334 Subcellular fractionation 1310:1334 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 2 23 theme intracellular 431:443 arg1 processing 461:470 an intracellular endoproteolytic processing 428:470 an intracellular endoproteolytic processing 428:470 Recently, we have shown that myocilin undergoes an intracellular endoproteolytic processing. 17650508 6 24 theme myocilin 986:993 arg1 cleavage 974:981 the cleavage 970:981 the cleavage of myocilin 970:993 Since calpains are calcium-activated proteases, we analyzed how changes in either intra- or extracellular calcium affected the cleavage of myocilin. 17650508 10 25 theme calpain 1510:1516 arg1 subpopulation 1493:1505 a subpopulation 1491:1505 a subpopulation of calpain II 1491:1519 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 9 26 theme RNA 1240:1242 arg1 knockdown 1257:1265 only RNA interference knockdown 1235:1265 only RNA interference knockdown of calpain II 1235:1279 However, in cells in culture, only RNA interference knockdown of calpain II reduced myocilin processing. 17650508 11 27 theme myocilin 1609:1616 arg1 processing 1595:1604 the intracellular processing 1577:1604 the intracellular processing of myocilin in the lumen of the endoplasmic reticulum 1577:1658 These data revealed that calpain II is responsible for the intracellular processing of myocilin in the lumen of the endoplasmic reticulum. 17650508 10 28 theme fractions 1366:1374 arg1 digestion 1340:1348 digestion 1340:1348 digestion 1340:1348 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 10 28 theme fractions 1366:1374 arg1 fractionation 1322:1334 Subcellular fractionation 1310:1334 Subcellular fractionation 1310:1334 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 1 29 theme glaucoma 183:190 arg1 types 174:178 different types 164:178 different types of glaucoma 164:190 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 0 30 theme intracellular 24:36 arg1 cleavage 50:57 the intracellular proteolytic cleavage 20:57 the intracellular proteolytic cleavage of myocilin 20:69 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 10 31 theme endoplasmic 1455:1465 arg1 reticulum 1467:1475 the endoplasmic reticulum 1451:1475 the endoplasmic reticulum together with a subpopulation of calpain II 1451:1519 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 5 32 theme transfected 715:725 arg1 cells 732:736 transiently transfected 293T cells 703:736 transiently transfected 293T cells 703:736 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 9 33 theme interference 1244:1255 arg1 knockdown 1257:1265 only RNA interference knockdown 1235:1265 only RNA interference knockdown of calpain II 1235:1279 However, in cells in culture, only RNA interference knockdown of calpain II reduced myocilin processing. 17650508 3 34 theme terminal 554:561 arg1 domains 563:569 the two terminal domains 546:569 the two terminal domains 546:569 We show herein that the proteolytic cleavage in the linker region splits the two terminal domains. 17650508 10 35 theme obtained 1357:1364 arg1 fractions 1366:1374 the obtained fractions 1353:1374 the obtained fractions 1353:1374 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 1 36 theme N-terminal 273:282 arg1 domain 304:309 an N-terminal leucine-zipper-like domain 270:309 an N-terminal leucine-zipper-like domain 270:309 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 5 37 theme 293T 727:730 arg1 cells 732:736 transiently transfected 293T cells 703:736 transiently transfected 293T cells 703:736 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 0 38 theme cleavage 50:57 arg1 Characterization 0:15 Characterization 0:15 Characterization of the intracellular proteolytic cleavage of myocilin 0:69 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 0 38 theme cleavage 50:57 arg1 identification 75:88 identification 75:88 identification of calpain II as a myocilin-processing protease 75:136 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 1 39 theme leucine-zipper-like 284:302 arg1 domain 304:309 an N-terminal leucine-zipper-like domain 270:309 an N-terminal leucine-zipper-like domain 270:309 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 7 40 theme extracellular 1092:1104 arg1 calcium 1106:1112 extracellular calcium 1092:1112 extracellular calcium 1092:1112 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 6 41 theme calcium-activated 866:882 arg1 calpains 853:860 calpains 853:860 calpains 853:860 Since calpains are calcium-activated proteases, we analyzed how changes in either intra- or extracellular calcium affected the cleavage of myocilin. 17650508 6 41 theme calcium-activated 866:882 arg1 proteases 884:892 calcium-activated proteases 866:892 calcium-activated proteases 866:892 Since calpains are calcium-activated proteases, we analyzed how changes in either intra- or extracellular calcium affected the cleavage of myocilin. 17650508 4 42 theme N-terminal 641:650 arg1 domain 652:657 the N-terminal domain 637:657 the N-terminal domain 637:657 The C-terminal domain is secreted to the culture medium, whereas the N-terminal domain mainly remains intracellularly retained. 17650508 0 43 theme proteolytic 38:48 arg1 cleavage 50:57 the intracellular proteolytic cleavage 20:57 the intracellular proteolytic cleavage of myocilin 20:69 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 4 44 theme C-terminal 576:585 arg1 domain 587:592 The C-terminal domain 572:592 The C-terminal domain 572:592 The C-terminal domain is secreted to the culture medium, whereas the N-terminal domain mainly remains intracellularly retained. 17650508 9 45 theme calpain 1270:1276 arg1 knockdown 1257:1265 only RNA interference knockdown 1235:1265 only RNA interference knockdown of calpain II 1235:1279 However, in cells in culture, only RNA interference knockdown of calpain II reduced myocilin processing. 17650508 7 46 theme myocilin 1052:1059 arg1 cleavage 1061:1068 myocilin cleavage 1052:1068 myocilin cleavage 1052:1068 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 0 47 theme myocilin 62:69 arg1 cleavage 50:57 the intracellular proteolytic cleavage 20:57 the intracellular proteolytic cleavage of myocilin 20:69 Characterization of the intracellular proteolytic cleavage of myocilin and identification of calpain II as a myocilin-processing protease. 17650508 6 48 from changes 911:917 arg1 extracellular 939:951 extracellular 939:951 extracellular 939:951 Since calpains are calcium-activated proteases, we analyzed how changes in either intra- or extracellular calcium affected the cleavage of myocilin. 17650508 6 48 from changes 911:917 arg1 intra- 929:934 intra- 929:934 intra- 929:934 Since calpains are calcium-activated proteases, we analyzed how changes in either intra- or extracellular calcium affected the cleavage of myocilin. 17650508 11 49 theme reticulum 1650:1658 arg1 lumen 1625:1629 the lumen 1621:1629 the lumen of the endoplasmic reticulum 1621:1658 These data revealed that calpain II is responsible for the intracellular processing of myocilin in the lumen of the endoplasmic reticulum. 17650508 7 50 theme calcium 1106:1112 arg1 chelation 1079:1087 chelation 1079:1087 chelation of extracellular calcium by EGTA 1079:1120 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 10 51 theme proteinase 1381:1390 arg1 K 1392:1392 proteinase K 1381:1392 proteinase K 1381:1392 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 3 52 theme proteolytic 497:507 arg1 cleavage 509:516 the proteolytic cleavage 493:516 the proteolytic cleavage in the linker region 493:537 We show herein that the proteolytic cleavage in the linker region splits the two terminal domains. 17650508 1 53 theme central 314:320 arg1 region 329:334 a central linker region 312:334 a central linker region 312:334 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 9 54 theme myocilin 1289:1296 arg1 processing 1298:1307 myocilin processing 1289:1307 myocilin processing 1289:1307 However, in cells in culture, only RNA interference knockdown of calpain II reduced myocilin processing. 17650508 11 55 theme endoplasmic 1638:1648 arg1 reticulum 1650:1658 the endoplasmic reticulum 1634:1658 the endoplasmic reticulum 1634:1658 These data revealed that calpain II is responsible for the intracellular processing of myocilin in the lumen of the endoplasmic reticulum. 17650508 7 56 theme ionomycin-induced 1010:1026 arg1 uptake 1036:1041 Intracellular ionomycin-induced calcium uptake 996:1041 Intracellular ionomycin-induced calcium uptake 996:1041 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 12 57 theme pressure 1789:1796 arg1 control 1766:1772 the control 1762:1772 the control of intraocular pressure 1762:1796 We propose that this cleavage might regulate extracellular interactions of myocilin, contributing to the control of intraocular pressure. 17650508 1 58 theme secreted 213:220 arg1 glycoprotein 222:233 a secreted glycoprotein 211:233 a secreted glycoprotein of unknown function 211:253 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 1 58 theme secreted 213:220 arg1 myocilin 201:208 myocilin 201:208 myocilin 201:208 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 11 59 theme intracellular 1581:1593 arg1 processing 1595:1604 the intracellular processing 1577:1604 the intracellular processing of myocilin in the lumen of the endoplasmic reticulum 1577:1658 These data revealed that calpain II is responsible for the intracellular processing of myocilin in the lumen of the endoplasmic reticulum. 17650508 7 60 theme calcium 1028:1034 arg1 uptake 1036:1041 Intracellular ionomycin-induced calcium uptake 996:1041 Intracellular ionomycin-induced calcium uptake 996:1041 Intracellular ionomycin-induced calcium uptake enhanced myocilin cleavage, whereas chelation of extracellular calcium by EGTA inhibited the proteolytic processing. 17650508 12 61 theme intraocular 1777:1787 arg1 pressure 1789:1796 intraocular pressure 1777:1796 intraocular pressure 1777:1796 We propose that this cleavage might regulate extracellular interactions of myocilin, contributing to the control of intraocular pressure. 17650508 5 62 theme calpain 769:775 arg1 calpeptin 797:805 calpeptin 797:805 calpeptin 797:805 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 5 62 theme calpain 769:775 arg1 inhibitor 816:824 calpain inhibitor IV 808:827 calpain inhibitor IV 808:827 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 5 62 theme calpain 769:775 arg1 inhibitors 777:786 calpain inhibitors 769:786 calpain inhibitors 769:786 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 5 62 theme calpain 769:775 arg1 calpastatin 834:844 calpastatin 834:844 calpastatin 834:844 In transiently transfected 293T cells, the cleavage was prevented by calpain inhibitors, such as calpeptin, calpain inhibitor IV, and calpastatin. 17650508 1 63 theme linker 322:327 arg1 region 329:334 a central linker region 312:334 a central linker region 312:334 MYOC, a gene involved in different types of glaucoma, encodes myocilin, a secreted glycoprotein of unknown function, consisting of an N-terminal leucine-zipper-like domain, a central linker region, and a C-terminal olfactomedin-like domain. 17650508 10 64 theme full-length 1406:1416 arg1 myocilin 1418:1425 full-length myocilin 1406:1425 full-length myocilin 1406:1425 Subcellular fractionation and digestion of the obtained fractions with proteinase K showed that full-length myocilin resides in the lumen of the endoplasmic reticulum together with a subpopulation of calpain II. 17650508 3 65 theme linker 525:530 arg1 region 532:537 the linker region 521:537 the linker region 521:537 We show herein that the proteolytic cleavage in the linker region splits the two terminal domains. 17650508 4 66 theme culture 613:619 arg1 medium 621:626 the culture medium 609:626 the culture medium 609:626 The C-terminal domain is secreted to the culture medium, whereas the N-terminal domain mainly remains intracellularly retained. 11081633 4 0 theme PSGL-1 761:766 arg1 domain 745:750 the N-terminal domain 730:750 the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X) 730:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 4 0 theme PSGL-1 761:766 arg1 PSGL-1 761:766 human PSGL-1 755:766 human PSGL-1 modified by both tyrosine sulfation and SLe(X) 755:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 1 1 theme adhesion 191:198 arg1 receptors 200:208 cell adhesion receptors 186:208 cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues 186:362 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 2 2 theme specific 457:464 arg1 counterreceptors 479:494 high-affinity to specific glycoprotein counterreceptors 440:494 high-affinity to specific glycoprotein counterreceptors 440:494 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 2 2 theme specific 457:464 arg1 PSGL-1 507:512 PSGL-1 507:512 PSGL-1 507:512 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 4 3 theme human 755:759 arg1 PSGL-1 761:766 human PSGL-1 755:766 human PSGL-1 modified by both tyrosine sulfation and SLe(X) 755:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 1 4 theme receptors 200:208 arg1 family 176:181 a family 174:181 a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues 174:362 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 3 5 theme EGF 614:616 arg1 domains 623:629 the lectin and EGF (LE) domains 599:629 the lectin and EGF (LE) domains co-complexed with SLe(X) 599:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 1 6 from rolling 249:255 arg1 endothelium 283:293 inflamed endothelium 274:293 inflamed endothelium 274:293 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 3 7 theme co-complexed 631:642 arg1 domains 623:629 the lectin and EGF (LE) domains 599:629 the lectin and EGF (LE) domains co-complexed with SLe(X) 599:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 1 8 from tethering 235:243 arg1 endothelium 283:293 inflamed endothelium 274:293 inflamed endothelium 274:293 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 3 9 contain containing 588:597 arg2 domains 623:629 the lectin and EGF (LE) domains 599:629 the lectin and EGF (LE) domains co-complexed with SLe(X) 599:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 3 9 contain containing 588:597 arg1 constructs 577:586 human P- and E-selectin constructs 553:586 human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X) 553:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 0 10 theme P- 95:96 arg1 structures 81:90 structures 81:90 structures of P- and E-selectin 81:111 Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. 11081633 2 11 theme sialyl 394:399 arg1 SLe 409:411 SLe 409:411 SLe(X) 409:414 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 2 11 theme sialyl 394:399 arg1 Lewisx 401:406 sialyl Lewisx 394:406 sialyl Lewisx (SLe(X)) 394:415 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 1 12 theme firm 317:320 arg1 attachment 322:331 their firm attachment 311:331 their firm attachment 311:331 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 2 13 theme -like 416:420 arg1 glycans 422:428 sialyl Lewisx (SLe(X))-like glycans 394:428 sialyl Lewisx (SLe(X))-like glycans 394:428 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 4 14 theme N-terminal 734:743 arg1 domain 745:750 the N-terminal domain 730:750 the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X) 730:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 4 14 theme N-terminal 734:743 arg1 PSGL-1 761:766 human PSGL-1 755:766 human PSGL-1 modified by both tyrosine sulfation and SLe(X) 755:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 5 15 theme high-affinity 920:932 arg1 interaction 934:944 the high-affinity interaction 916:944 the high-affinity interaction between P-selectin and PSGL-1 916:974 These structures reveal differences in how E- and P-selectin bind SLe(X) and the molecular basis of the high-affinity interaction between P-selectin and PSGL-1. 11081633 0 16 theme molecular 18:26 arg1 basis 28:32 the molecular basis 14:32 the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin 14:111 Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. 11081633 1 17 theme initial 227:233 arg1 tethering 235:243 initial tethering 227:243 initial tethering 227:243 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 1 17 theme initial 227:233 arg1 rolling 249:255 rolling 249:255 rolling 249:255 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 1 17 theme initial 227:233 arg1 prelude 300:306 a prelude 298:306 a prelude to their firm attachment and extravasation into tissues 298:362 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 2 18 theme glycoprotein 466:477 arg1 counterreceptors 479:494 high-affinity to specific glycoprotein counterreceptors 440:494 high-affinity to specific glycoprotein counterreceptors 440:494 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 2 18 theme glycoprotein 466:477 arg1 PSGL-1 507:512 PSGL-1 507:512 PSGL-1 507:512 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 0 19 theme E-selectin 102:111 arg1 structures 81:90 structures 81:90 structures of P- and E-selectin 81:111 Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. 11081633 3 20 theme E-selectin 566:575 arg1 constructs 577:586 human P- and E-selectin constructs 553:586 human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X) 553:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 4 21 theme LE 709:710 arg1 structure 685:693 the crystal structure 673:693 the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X) 673:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 2 22 theme high-affinity 440:452 arg1 counterreceptors 479:494 high-affinity to specific glycoprotein counterreceptors 440:494 high-affinity to specific glycoprotein counterreceptors 440:494 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 2 22 theme high-affinity 440:452 arg1 PSGL-1 507:512 PSGL-1 507:512 PSGL-1 507:512 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 3 23 theme human 553:557 arg1 constructs 577:586 human P- and E-selectin constructs 553:586 human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X) 553:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 4 24 theme P-selectin 698:707 arg1 LE 709:710 P-selectin LE 698:710 P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X) 698:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 0 25 theme tethering 47:55 arg1 basis 28:32 the molecular basis 14:32 the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin 14:111 Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. 11081633 4 26 theme tyrosine 785:792 arg1 sulfation 794:802 tyrosine sulfation 785:802 tyrosine sulfation 785:802 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 3 27 theme P- 559:560 arg1 constructs 577:586 human P- and E-selectin constructs 553:586 human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X) 553:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 0 28 theme leukocyte 37:45 arg1 tethering 47:55 leukocyte tethering 37:55 leukocyte tethering 37:55 Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. 11081633 1 29 theme leukocytes 260:269 arg1 tethering 235:243 initial tethering 227:243 initial tethering 227:243 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 1 29 theme leukocytes 260:269 arg1 prelude 300:306 a prelude 298:306 a prelude to their firm attachment and extravasation into tissues 298:362 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 1 29 theme leukocytes 260:269 arg1 rolling 249:255 rolling 249:255 rolling 249:255 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 1 30 dep tethering 235:243 arg1 the 223:225 the 223:225 the 223:225 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 5 31 theme molecular 897:905 arg1 basis 907:911 the molecular basis 893:911 the molecular basis of the high-affinity interaction between P-selectin and PSGL-1 893:974 These structures reveal differences in how E- and P-selectin bind SLe(X) and the molecular basis of the high-affinity interaction between P-selectin and PSGL-1. 11081633 5 32 theme interaction 934:944 arg1 basis 907:911 the molecular basis 893:911 the molecular basis of the high-affinity interaction between P-selectin and PSGL-1 893:974 These structures reveal differences in how E- and P-selectin bind SLe(X) and the molecular basis of the high-affinity interaction between P-selectin and PSGL-1. 11081633 5 32 theme interaction 934:944 arg1 X 886:886 X 886:886 X 886:886 These structures reveal differences in how E- and P-selectin bind SLe(X) and the molecular basis of the high-affinity interaction between P-selectin and PSGL-1. 11081633 5 32 theme interaction 934:944 arg1 SLe 882:884 SLe 882:884 SLe(X) 882:887 These structures reveal differences in how E- and P-selectin bind SLe(X) and the molecular basis of the high-affinity interaction between P-selectin and PSGL-1. 11081633 3 33 theme lectin 603:608 arg1 domains 623:629 the lectin and EGF (LE) domains 599:629 the lectin and EGF (LE) domains co-complexed with SLe(X) 599:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 4 34 theme crystal 677:683 arg1 structure 685:693 the crystal structure 673:693 the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X) 673:813 We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). 11081633 3 35 theme constructs 577:586 arg1 structures 539:548 crystal structures 531:548 crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X) 531:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 3 36 with co-complexed 631:642 arg1 X 653:653 X 653:653 X 653:653 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 3 36 with co-complexed 631:642 arg1 SLe 649:651 SLe 649:651 SLe(X) 649:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 0 37 theme rolling 61:67 arg1 basis 28:32 the molecular basis 14:32 the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin 14:111 Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. 11081633 2 38 dep counterreceptors 479:494 arg1 to 454:455 to 454:455 to 454:455 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 1 39 theme inflamed 274:281 arg1 endothelium 283:293 inflamed endothelium 274:293 inflamed endothelium 274:293 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 11081633 3 40 theme crystal 531:537 arg1 structures 539:548 crystal structures 531:548 crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X) 531:654 Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). 11081633 2 41 gly glycoprotein 466:477 arg1 glycoprotein 466:477 high-affinity to specific glycoprotein counterreceptors 440:494 high-affinity to specific glycoprotein counterreceptors 440:494 The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. 11081633 1 42 theme cell 186:189 arg1 receptors 200:208 cell adhesion receptors 186:208 cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues 186:362 P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. 9883900 0 0 theme growth 96:101 arg1 protein-5 118:126 human insulin-like growth factor binding protein-5 77:126 human insulin-like growth factor binding protein-5 77:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 4 1 from kDa 617:619 arg1 sizes 597:601 molecular sizes 587:601 molecular sizes from 12 to 25 kDa 587:619 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. 9883900 5 2 theme IGFBP-5 661:667 arg1 peptides 649:656 C-terminal peptides 638:656 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa 638:710 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 9883900 10 3 theme IGFBP-5 1385:1391 arg1 proteolysis 1370:1380 plasma proteolysis 1363:1380 plasma proteolysis of IGFBP-5 1363:1391 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 2 4 theme human 357:361 arg1 hemofiltrate 363:374 human hemofiltrate 357:374 human hemofiltrate 357:374 We screened for circulating fragments of human IGFBP-5 in human hemofiltrate. 9883900 0 5 theme insulin-like 83:94 arg1 protein-5 118:126 human insulin-like growth factor binding protein-5 77:126 human insulin-like growth factor binding protein-5 77:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 3 6 from fractions 419:427 arg1 Identification 377:390 Identification 377:390 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate 377:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 10 7 theme IGF-dependent 1514:1526 arg1 manner 1528:1533 an IGF-dependent manner 1511:1533 an IGF-dependent manner 1511:1533 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 1 8 theme insulin-like 263:274 arg1 IGFs 292:295 IGFs 292:295 IGFs 292:295 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 1 8 theme insulin-like 263:274 arg1 factors 283:289 the insulin-like growth factors 259:289 the insulin-like growth factors (IGFs) 259:296 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 0 9 theme binding 110:116 arg1 protein-5 118:126 human insulin-like growth factor binding protein-5 77:126 human insulin-like growth factor binding protein-5 77:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 7 10 theme central 935:941 arg1 region 951:956 the central IGFBP-5 region 931:956 the central IGFBP-5 region 931:956 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 5 11 theme molecular 674:682 arg1 masses 684:689 molecular masses 674:689 molecular masses of 13.3 and 13.5 kDa 674:710 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 9883900 6 12 theme double 826:831 arg1 K 845:845 K 845:845 K 845:845 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 6 12 theme double 826:831 arg1 sequences 834:842 the (double) sequences 821:842 the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG 821:880 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 6 12 theme double 826:831 arg1 MVPRAVYLPNXDRKG 866:880 MVPRAVYLPNXDRKG 866:880 MVPRAVYLPNXDRKG 866:880 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 6 12 theme double 826:831 arg1 FVGGAENXAHPRII 847:860 FVGGAENXAHPRII 847:860 FVGGAENXAHPRII 847:860 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 1 13 theme growth 276:281 arg1 IGFs 292:295 IGFs 292:295 IGFs 292:295 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 1 13 theme growth 276:281 arg1 factors 283:289 the insulin-like growth factors 259:289 the insulin-like growth factors (IGFs) 259:296 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 0 14 theme factor 103:108 arg1 protein-5 118:126 human insulin-like growth factor binding protein-5 77:126 human insulin-like growth factor binding protein-5 77:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 3 15 from Identification 377:390 arg1 fractions 419:427 the fractions 415:427 the fractions of our peptide bank generated from hemofiltrate 415:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 7 16 theme IGFBP-5 943:949 arg1 region 951:956 the central IGFBP-5 region 931:956 the central IGFBP-5 region 931:956 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 10 17 theme biological 1557:1566 arg1 functions 1568:1576 intrinsic biological functions 1547:1576 intrinsic biological functions 1547:1576 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 1 18 theme factors 283:289 arg1 bioavailability 240:254 the bioavailability 236:254 the bioavailability of the insulin-like growth factors (IGFs) 236:296 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 1 18 theme factors 283:289 arg1 regulation 206:215 regulation 206:215 regulation of the effects 206:230 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 5 19 with peptides 649:656 arg1 masses 684:689 molecular masses 674:689 molecular masses of 13.3 and 13.5 kDa 674:710 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 9883900 7 20 theme region 951:956 arg1 2722 923:926 Mr 2722 920:926 Mr 2722 of the central IGFBP-5 region 920:956 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 3 21 theme bank 444:447 arg1 fractions 419:427 the fractions 415:427 the fractions of our peptide bank generated from hemofiltrate 415:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 0 22 theme protein-5 118:126 arg1 fragments 64:72 circulating 13-kDa C-terminal fragments 34:72 circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5 34:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 5 23 theme C-terminal 638:647 arg1 peptides 649:656 C-terminal peptides 638:656 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa 638:710 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 9883900 8 24 theme sequence 1181:1188 arg1 analysis 1190:1197 sequence analysis 1181:1197 sequence analysis 1181:1197 According to mass spectrometric and sequence analysis, Thr-152 was shown to be O-glycosylated. 9883900 7 25 dep residues 1020:1027 arg1 121-143 1029:1035 121-143 1029:1035 121-143 1029:1035 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 3 26 theme IGFBP-5 395:401 arg1 peptides 403:410 IGFBP-5 peptides 395:410 IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate 395:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 3 27 from peptides 403:410 arg1 fractions 419:427 the fractions 415:427 the fractions of our peptide bank generated from hemofiltrate 415:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 7 28 theme amino 1093:1097 arg1 acids 1099:1103 amino acids Lys-120, Ser-143, Lys-144, and Arg-188 1093:1142 amino acids Lys-120, Ser-143, Lys-144, and Arg-188 1093:1142 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 6 29 theme peptides 803:810 arg1 analysis 787:794 Sequence analysis 778:794 Sequence analysis of the peptides 778:810 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 4 30 with fragments 561:569 arg1 sizes 597:601 molecular sizes 587:601 molecular sizes from 12 to 25 kDa 587:619 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. 9883900 7 31 with fragment 906:913 arg1 2722 923:926 Mr 2722 920:926 Mr 2722 of the central IGFBP-5 region 920:956 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 3 32 theme peptides 403:410 arg1 Identification 377:390 Identification 377:390 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate 377:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 4 33 theme IGFBP-5 574:580 arg1 fragments 561:569 Different fragments 551:569 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa 551:619 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. 9883900 10 34 theme different 1460:1468 arg1 fragments 1481:1489 different C-terminal fragments 1460:1489 different C-terminal fragments 1460:1489 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 10 35 theme basic 1431:1435 arg1 residues 1437:1444 basic residues 1431:1444 basic residues 1431:1444 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 3 36 theme peptide 436:442 arg1 bank 444:447 our peptide bank 432:447 our peptide bank generated from hemofiltrate 432:475 Identification of IGFBP-5 peptides in the fractions of our peptide bank generated from hemofiltrate was performed by their immunoreactivity and their capacity to bind IGF-I. 9883900 6 37 dep sequences 834:842 arg1 sequences 834:842 the (double) sequences 821:842 the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG 821:880 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 6 37 dep sequences 834:842 arg1 MVPRAVYLPNXDRKG 866:880 MVPRAVYLPNXDRKG 866:880 MVPRAVYLPNXDRKG 866:880 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 6 37 dep sequences 834:842 arg1 FVGGAENXAHPRII 847:860 FVGGAENXAHPRII 847:860 FVGGAENXAHPRII 847:860 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 5 38 theme kDa 708:710 arg1 masses 684:689 molecular masses 674:689 molecular masses of 13.3 and 13.5 kDa 674:710 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 9883900 4 39 theme molecular 587:595 arg1 sizes 597:601 molecular sizes 587:601 molecular sizes from 12 to 25 kDa 587:619 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. 9883900 10 40 theme plasma 1363:1368 arg1 proteolysis 1370:1380 plasma proteolysis 1363:1380 plasma proteolysis of IGFBP-5 1363:1391 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 8 41 gly O-glycosylated 1224:1237 arg1 Thr-152 1200:1206 Thr-152 1200:1206 Thr-152 1200:1206 According to mass spectrometric and sequence analysis, Thr-152 was shown to be O-glycosylated. 9883900 10 42 theme C-terminal 1470:1479 arg1 fragments 1481:1489 different C-terminal fragments 1460:1489 different C-terminal fragments 1460:1489 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 0 43 theme 13-kDa 46:51 arg1 fragments 64:72 circulating 13-kDa C-terminal fragments 34:72 circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5 34:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 2 44 theme human 340:344 arg1 IGFBP-5 346:352 human IGFBP-5 340:352 human IGFBP-5 340:352 We screened for circulating fragments of human IGFBP-5 in human hemofiltrate. 9883900 10 45 theme intrinsic 1547:1555 arg1 functions 1568:1576 intrinsic biological functions 1547:1576 intrinsic biological functions 1547:1576 Our results indicate that plasma proteolysis of IGFBP-5 preferentially occurs C-terminally to basic residues and generates different C-terminal fragments, possibly acting in an IGF-dependent manner and bearing intrinsic biological functions. 9883900 0 46 theme circulating 34:44 arg1 fragments 64:72 circulating 13-kDa C-terminal fragments 34:72 circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5 34:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 7 47 theme IGFBP-5 1071:1077 arg1 proteolysis 1056:1066 plasma proteolysis 1049:1066 plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188 1049:1142 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 1 48 theme effects 224:230 arg1 bioavailability 240:254 the bioavailability 236:254 the bioavailability of the insulin-like growth factors (IGFs) 236:296 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 1 48 theme effects 224:230 arg1 regulation 206:215 regulation 206:215 regulation of the effects 206:230 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 0 49 theme fragments 64:72 arg1 characterization 14:29 characterization 14:29 characterization 14:29 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 0 49 theme fragments 64:72 arg1 Isolation 0:8 Isolation 0:8 Isolation 0:8 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 5 50 theme consecutive 729:739 arg1 steps 757:761 consecutive chromatographic steps 729:761 consecutive chromatographic steps 729:761 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 9883900 7 51 theme plasma 1049:1054 arg1 proteolysis 1056:1066 plasma proteolysis 1049:1066 plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188 1049:1142 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 6 52 theme Sequence 778:785 arg1 analysis 787:794 Sequence analysis 778:794 Sequence analysis of the peptides 778:810 Sequence analysis of the peptides revealed the (double) sequences (K)FVGGAENXAHPRII and MVPRAVYLPNXDRKG. 9883900 2 53 theme IGFBP-5 346:352 arg1 fragments 327:335 fragments 327:335 fragments of human IGFBP-5 327:352 We screened for circulating fragments of human IGFBP-5 in human hemofiltrate. 9883900 0 54 theme C-terminal 53:62 arg1 fragments 64:72 circulating 13-kDa C-terminal fragments 34:72 circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5 34:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 9 55 theme significant 1299:1309 arg1 properties 1325:1334 significant IGF-I binding properties 1299:1334 significant IGF-I binding properties 1299:1334 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 9 56 theme C-terminal 1261:1270 arg1 fragments 1280:1288 C-terminal IGFBP-5 fragments 1261:1288 C-terminal IGFBP-5 fragments 1261:1288 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 9 57 theme IGF-I 1311:1315 arg1 properties 1325:1334 significant IGF-I binding properties 1299:1334 significant IGF-I binding properties 1299:1334 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 1 58 theme insulin-like 133:144 arg1 factor 153:158 The insulin-like growth factor 129:158 The insulin-like growth factor binding proteins (IGFBPs) 129:184 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 9 59 contain containing 1250:1259 arg1 Fractions 1240:1248 Fractions 1240:1248 Fractions containing C-terminal IGFBP-5 fragments 1240:1288 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 9 59 contain containing 1250:1259 arg2 fragments 1280:1288 C-terminal IGFBP-5 fragments 1261:1288 C-terminal IGFBP-5 fragments 1261:1288 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 4 60 theme Different 551:559 arg1 fragments 561:569 Different fragments 551:569 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa 551:619 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. 9883900 9 61 theme IGFBP-5 1272:1278 arg1 fragments 1280:1288 C-terminal IGFBP-5 fragments 1261:1288 C-terminal IGFBP-5 fragments 1261:1288 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 1 62 theme growth 146:151 arg1 factor 153:158 The insulin-like growth factor 129:158 The insulin-like growth factor binding proteins (IGFBPs) 129:184 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 7 63 theme Mr 920:921 arg1 2722 923:926 Mr 2722 920:926 Mr 2722 of the central IGFBP-5 region 920:956 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 8 64 theme mass 1158:1161 arg1 spectrometric 1163:1175 mass spectrometric 1158:1175 mass spectrometric 1158:1175 According to mass spectrometric and sequence analysis, Thr-152 was shown to be O-glycosylated. 9883900 1 65 theme factor 153:158 arg1 proteins 168:175 The insulin-like growth factor binding proteins 129:175 The insulin-like growth factor binding proteins (IGFBPs) 129:184 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 1 65 theme factor 153:158 arg1 IGFBPs 178:183 IGFBPs 178:183 IGFBPs 178:183 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 7 66 theme smaller 898:904 arg1 fragment 906:913 a smaller fragment 896:913 a smaller fragment with Mr 2722 of the central IGFBP-5 region 896:956 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 0 67 theme human 77:81 arg1 protein-5 118:126 human insulin-like growth factor binding protein-5 77:126 human insulin-like growth factor binding protein-5 77:126 Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5. 9883900 4 68 dep 25 614:615 arg1 to 611:612 to 611:612 to 611:612 Different fragments of IGFBP-5 with molecular sizes from 12 to 25 kDa were identified. 9883900 7 69 theme sequence 986:993 arg1 residues 1020:1027 residues 121-143 1020:1035 residues 121-143 1020:1035 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 7 69 theme sequence 986:993 arg1 HTRISELKAEAVKKDRRKKLTQS 995:1017 the sequence HTRISELKAEAVKKDRRKKLTQS 982:1017 the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) 982:1036 In addition, a smaller fragment with Mr 2722 of the central IGFBP-5 region was purified and showed the sequence HTRISELKAEAVKKDRRKKLTQS (residues 121-143) indicating plasma proteolysis of IGFBP-5 C-terminal to amino acids Lys-120, Ser-143, Lys-144, and Arg-188. 9883900 1 70 theme binding 160:166 arg1 proteins 168:175 The insulin-like growth factor binding proteins 129:175 The insulin-like growth factor binding proteins (IGFBPs) 129:184 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 1 70 theme binding 160:166 arg1 IGFBPs 178:183 IGFBPs 178:183 IGFBPs 178:183 The insulin-like growth factor binding proteins (IGFBPs) are responsible for regulation of the effects and the bioavailability of the insulin-like growth factors (IGFs). 9883900 9 71 theme binding 1317:1323 arg1 properties 1325:1334 significant IGF-I binding properties 1299:1334 significant IGF-I binding properties 1299:1334 Fractions containing C-terminal IGFBP-5 fragments revealed significant IGF-I binding properties. 9883900 5 72 theme chromatographic 741:755 arg1 steps 757:761 consecutive chromatographic steps 729:761 consecutive chromatographic steps 729:761 C-terminal peptides of IGFBP-5 with molecular masses of 13.3 and 13.5 kDa were purified by consecutive chromatographic steps and sequenced. 19122660 0 0 theme domain 70:75 arg1 tunnel 32:37 the tunnel 28:37 the tunnel of a ten-bladed beta-propeller domain 28:75 Ligands bind to Sortilin in the tunnel of a ten-bladed beta-propeller domain. 19122660 2 1 theme binding 347:353 arg1 sites 355:359 additional binding sites 336:359 additional binding sites 336:359 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 1 2 theme neurotensin 224:234 arg1 part 216:219 the C-terminal part 201:219 the C-terminal part of neurotensin 201:234 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 0 3 theme beta-propeller 55:68 arg1 domain 70:75 a ten-bladed beta-propeller domain 42:75 a ten-bladed beta-propeller domain 42:75 Ligands bind to Sortilin in the tunnel of a ten-bladed beta-propeller domain. 19122660 2 4 theme additional 336:345 arg1 sites 355:359 additional binding sites 336:359 additional binding sites 336:359 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 1 5 with complex 122:128 arg1 neurotensin 135:145 neurotensin 135:145 neurotensin 135:145 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 2 6 located present 426:432 arg2 sites 355:359 additional binding sites 336:359 additional binding sites 336:359 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 6 located present 426:432 arg1 tunnel 441:446 the tunnel 437:446 the tunnel 437:446 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 7 attach present 426:432 arg2 sites 355:359 additional binding sites 336:359 additional binding sites 336:359 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 7 attach present 426:432 arg1 tunnel 441:446 the tunnel 437:446 the tunnel 437:446 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 8 from present 426:432 arg1 tunnel 441:446 the tunnel 437:446 the tunnel 437:446 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 9 from tunnel 441:446 arg1 present 426:432 present 426:432 present 426:432 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 10 theme Binding 295:301 arg1 studies 315:321 Binding competition studies 295:321 Binding competition studies 295:321 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 1 11 theme 2-A 170:172 arg1 resolution 174:183 2-A resolution 170:183 2-A resolution 170:183 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 2 12 theme factor-beta 409:419 arg1 prodomain 383:391 the prodomain 379:391 the prodomain of nerve growth factor-beta 379:419 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 12 theme factor-beta 409:419 arg1 factor-beta 409:419 nerve growth factor-beta 396:419 nerve growth factor-beta 396:419 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 13 theme nerve 396:400 arg1 factor-beta 409:419 nerve growth factor-beta 396:419 nerve growth factor-beta 396:419 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 2 14 theme growth 402:407 arg1 factor-beta 409:419 nerve growth factor-beta 396:419 nerve growth factor-beta 396:419 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 1 15 theme ten-bladed 261:270 arg1 domain 287:292 a ten-bladed beta-propeller domain 259:292 a ten-bladed beta-propeller domain 259:292 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 1 16 theme beta-propeller 272:285 arg1 domain 287:292 a ten-bladed beta-propeller domain 259:292 a ten-bladed beta-propeller domain 259:292 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 2 17 theme restricted 496:505 arg1 space 507:511 the restricted space 492:511 the restricted space inside the propeller 492:532 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 1 18 theme domain 287:292 arg1 tunnel 249:254 the tunnel 245:254 the tunnel of a ten-bladed beta-propeller domain 245:292 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 1 19 theme Sortilin 99:106 arg1 ectodomain 108:117 the Sortilin ectodomain 95:117 the Sortilin ectodomain 95:117 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 1 20 from structure 82:90 arg1 complex 122:128 complex 122:128 complex with neurotensin 122:145 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 1 21 theme ectodomain 108:117 arg1 structure 82:90 The structure 78:90 The structure of the Sortilin ectodomain in complex with neurotensin 78:145 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 2 22 theme competition 303:313 arg1 studies 315:321 Binding competition studies 295:321 Binding competition studies 295:321 Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. 19122660 1 23 theme C-terminal 205:214 arg1 part 216:219 the C-terminal part 201:219 the C-terminal part of neurotensin 201:234 The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. 19122660 0 24 theme ten-bladed 44:53 arg1 domain 70:75 a ten-bladed beta-propeller domain 42:75 a ten-bladed beta-propeller domain 42:75 Ligands bind to Sortilin in the tunnel of a ten-bladed beta-propeller domain. 26091039 8 0 theme major 982:986 arg1 pathway 998:1004 the major secretory pathway protein kinase 978:1019 the major secretory pathway protein kinase 978:1019 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 8 0 theme major 982:986 arg1 Fam20C 968:973 Fam20C 968:973 Fam20C 968:973 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 5 1 theme genuine 646:652 arg1 phosphoproteins 627:641 more than 100 secreted phosphoproteins 604:641 more than 100 secreted phosphoproteins 604:641 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 5 1 theme genuine 646:652 arg1 substrates 661:670 genuine Fam20C substrates 646:670 genuine Fam20C substrates 646:670 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 5 2 theme genome 539:544 arg1 editing 546:552 CRISPR/Cas9 genome editing 527:552 CRISPR/Cas9 genome editing 527:552 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 7 3 theme cell 917:920 arg1 migration 922:930 cell migration 917:930 cell migration 917:930 Functional annotations of Fam20C substrates suggest roles for the kinase beyond biomineralization, including lipid homeostasis, wound healing, and cell migration and adhesion. 26091039 2 4 from research 170:177 arg1 area 187:190 this area 182:190 this area 182:190 However, research in this area has been undeveloped largely because the kinases that phosphorylate secreted proteins have escaped identification. 26091039 4 5 theme phosphoproteome 504:518 arg1 majority 474:481 the majority 470:481 the majority of the extracellular phosphoproteome 470:518 Here, we show that Fam20C generates the majority of the extracellular phosphoproteome. 26091039 5 6 theme Fam20C 654:659 arg1 phosphoproteins 627:641 more than 100 secreted phosphoproteins 604:641 more than 100 secreted phosphoproteins 604:641 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 5 6 theme Fam20C 654:659 arg1 substrates 661:670 genuine Fam20C substrates 646:670 genuine Fam20C substrates 646:670 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 8 7 theme protein 1006:1012 arg1 kinase 1014:1019 protein kinase 1006:1019 the major secretory pathway protein kinase 978:1019 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 8 8 theme new 1051:1053 arg1 areas 1055:1059 new areas 1051:1059 new areas of investigation into the role of secreted protein phosphorylation in human biology and disease 1051:1155 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 4 9 theme extracellular 490:502 arg1 phosphoproteome 504:518 the extracellular phosphoproteome 486:518 the extracellular phosphoproteome 486:518 Here, we show that Fam20C generates the majority of the extracellular phosphoproteome. 26091039 6 10 theme substrate 719:727 arg1 specificity 729:739 broader substrate specificity 711:739 broader substrate specificity 711:739 Further, we show that Fam20C exhibits broader substrate specificity than previously appreciated. 26091039 6 11 theme broader 711:717 arg1 specificity 729:739 broader substrate specificity 711:739 broader substrate specificity 711:739 Further, we show that Fam20C exhibits broader substrate specificity than previously appreciated. 26091039 7 12 theme wound 898:902 arg1 healing 904:910 wound healing 898:910 wound healing 898:910 Functional annotations of Fam20C substrates suggest roles for the kinase beyond biomineralization, including lipid homeostasis, wound healing, and cell migration and adhesion. 26091039 5 13 theme secreted 618:625 arg1 phosphoproteins 627:641 more than 100 secreted phosphoproteins 604:641 more than 100 secreted phosphoproteins 604:641 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 5 13 theme secreted 618:625 arg1 substrates 661:670 genuine Fam20C substrates 646:670 genuine Fam20C substrates 646:670 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 7 14 theme lipid 879:883 arg1 homeostasis 885:895 lipid homeostasis 879:895 lipid homeostasis 879:895 Functional annotations of Fam20C substrates suggest roles for the kinase beyond biomineralization, including lipid homeostasis, wound healing, and cell migration and adhesion. 26091039 0 15 theme Single 2:7 arg1 Kinase 9:14 A Single Kinase 0:14 A Single Kinase 0:14 A Single Kinase Generates the Majority of the Secreted Phosphoproteome. 26091039 3 16 theme teeth 427:431 arg1 matrix 407:412 the extracellular matrix 389:412 the extracellular matrix of bones and teeth 389:431 Fam20C is a kinase that phosphorylates S-x-E/pS motifs on proteins in milk and in the extracellular matrix of bones and teeth. 26091039 8 17 theme secreted 1095:1102 arg1 phosphorylation 1112:1126 secreted protein phosphorylation 1095:1126 secreted protein phosphorylation 1095:1126 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 1 18 theme extracellular 89:101 arg1 phosphoproteins 103:117 extracellular phosphoproteins 89:117 extracellular phosphoproteins 89:117 The existence of extracellular phosphoproteins has been acknowledged for over a century. 26091039 1 19 theme phosphoproteins 103:117 arg1 existence 76:84 The existence 72:84 The existence of extracellular phosphoproteins 72:117 The existence of extracellular phosphoproteins has been acknowledged for over a century. 26091039 8 20 theme phosphorylation 1112:1126 arg1 role 1087:1090 the role 1083:1090 the role of secreted protein phosphorylation in human biology and disease 1083:1155 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 8 21 dep pathway 998:1004 arg1 kinase 1014:1019 protein kinase 1006:1019 the major secretory pathway protein kinase 978:1019 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 3 22 theme bones 417:421 arg1 matrix 407:412 the extracellular matrix 389:412 the extracellular matrix of bones and teeth 389:431 Fam20C is a kinase that phosphorylates S-x-E/pS motifs on proteins in milk and in the extracellular matrix of bones and teeth. 26091039 7 23 theme substrates 803:812 arg1 annotations 781:791 Functional annotations 770:791 Functional annotations of Fam20C substrates 770:812 Functional annotations of Fam20C substrates suggest roles for the kinase beyond biomineralization, including lipid homeostasis, wound healing, and cell migration and adhesion. 26091039 8 24 theme protein 1104:1110 arg1 phosphorylation 1112:1126 secreted protein phosphorylation 1095:1126 secreted protein phosphorylation 1095:1126 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 3 25 from proteins 365:372 arg1 matrix 407:412 the extracellular matrix 389:412 the extracellular matrix of bones and teeth 389:431 Fam20C is a kinase that phosphorylates S-x-E/pS motifs on proteins in milk and in the extracellular matrix of bones and teeth. 26091039 3 25 from proteins 365:372 arg1 milk 377:380 milk 377:380 milk 377:380 Fam20C is a kinase that phosphorylates S-x-E/pS motifs on proteins in milk and in the extracellular matrix of bones and teeth. 26091039 3 26 theme extracellular 393:405 arg1 matrix 407:412 the extracellular matrix 389:412 the extracellular matrix of bones and teeth 389:431 Fam20C is a kinase that phosphorylates S-x-E/pS motifs on proteins in milk and in the extracellular matrix of bones and teeth. 26091039 8 27 from role 1087:1090 arg1 disease 1149:1155 disease 1149:1155 disease 1149:1155 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 8 27 from role 1087:1090 arg1 biology 1137:1143 human biology 1131:1143 human biology 1131:1143 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 8 28 theme human 1131:1135 arg1 biology 1137:1143 human biology 1131:1143 human biology 1131:1143 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 0 29 theme Secreted 46:53 arg1 Phosphoproteome 55:69 the Secreted Phosphoproteome 42:69 the Secreted Phosphoproteome 42:69 A Single Kinase Generates the Majority of the Secreted Phosphoproteome. 26091039 8 30 theme investigation 1064:1076 arg1 areas 1055:1059 new areas 1051:1059 new areas of investigation into the role of secreted protein phosphorylation in human biology and disease 1051:1155 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 5 31 theme mass 555:558 arg1 spectrometry 560:571 mass spectrometry 555:571 mass spectrometry 555:571 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 26091039 7 32 theme Fam20C 796:801 arg1 substrates 803:812 Fam20C substrates 796:812 Fam20C substrates 796:812 Functional annotations of Fam20C substrates suggest roles for the kinase beyond biomineralization, including lipid homeostasis, wound healing, and cell migration and adhesion. 26091039 2 33 theme secreted 260:267 arg1 proteins 269:276 secreted proteins 260:276 secreted proteins 260:276 However, research in this area has been undeveloped largely because the kinases that phosphorylate secreted proteins have escaped identification. 26091039 8 34 theme secretory 988:996 arg1 pathway 998:1004 the major secretory pathway protein kinase 978:1019 the major secretory pathway protein kinase 978:1019 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 8 34 theme secretory 988:996 arg1 Fam20C 968:973 Fam20C 968:973 Fam20C 968:973 Our results establish Fam20C as the major secretory pathway protein kinase and serve as a foundation for new areas of investigation into the role of secreted protein phosphorylation in human biology and disease. 26091039 7 35 theme Functional 770:779 arg1 annotations 781:791 Functional annotations 770:791 Functional annotations of Fam20C substrates 770:812 Functional annotations of Fam20C substrates suggest roles for the kinase beyond biomineralization, including lipid homeostasis, wound healing, and cell migration and adhesion. 26091039 0 36 theme Phosphoproteome 55:69 arg1 Majority 30:37 the Majority 26:37 the Majority of the Secreted Phosphoproteome 26:69 A Single Kinase Generates the Majority of the Secreted Phosphoproteome. 26091039 3 37 theme S-x-E/pS 346:353 arg1 motifs 355:360 S-x-E/pS motifs 346:360 S-x-E/pS motifs 346:360 Fam20C is a kinase that phosphorylates S-x-E/pS motifs on proteins in milk and in the extracellular matrix of bones and teeth. 26091039 5 38 theme CRISPR/Cas9 527:537 arg1 editing 546:552 CRISPR/Cas9 genome editing 527:552 CRISPR/Cas9 genome editing 527:552 Using CRISPR/Cas9 genome editing, mass spectrometry, and biochemistry, we identify more than 100 secreted phosphoproteins as genuine Fam20C substrates. 23851396 2 0 theme folate 433:438 arg1 conditions 440:449 low folate conditions 429:449 low folate conditions 429:449 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 1 1 theme cysteine-rich 118:130 arg1 receptors 85:93 Folate receptors 78:93 Folate receptors (FRα, FRβ and FRγ) 78:112 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 1 theme cysteine-rich 118:130 arg1 glycoproteins 145:157 cysteine-rich cell-surface glycoproteins 118:157 cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate 118:230 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 4 2 theme FRα 772:774 arg1 structure 753:761 the crystal structure 741:761 the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution 741:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 1 3 theme folate 225:230 arg1 uptake 215:220 cellular uptake 206:220 cellular uptake of folate 206:230 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 2 4 theme low 429:431 arg1 conditions 440:449 low folate conditions 429:449 low folate conditions 429:449 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 1 5 gly glycoproteins 145:157 arg1 receptors 85:93 Folate receptors 78:93 Folate receptors (FRα, FRβ and FRγ) 78:112 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 5 gly glycoproteins 145:157 arg1 glycoproteins 145:157 cysteine-rich cell-surface glycoproteins 118:157 cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate 118:230 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 6 dep receptors 85:93 arg1 receptors 85:93 Folate receptors 78:93 Folate receptors (FRα, FRβ and FRγ) 78:112 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 6 dep receptors 85:93 arg1 FRγ 109:111 FRγ 109:111 FRγ 109:111 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 6 dep receptors 85:93 arg1 FRβ 101:103 FRβ 101:103 FRβ 101:103 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 6 dep receptors 85:93 arg1 FRα 96:98 FRα 96:98 FRα 96:98 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 4 7 theme human 766:770 arg1 FRα 772:774 human FRα 766:774 human FRα 766:774 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 4 8 from structure 753:761 arg1 complex 779:785 complex 779:785 complex with folic acid at 2.8 Å resolution 779:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 4 9 theme crystal 745:751 arg1 structure 753:761 the crystal structure 741:761 the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution 741:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 5 10 theme deep 905:908 arg1 pocket 930:935 a deep open folate-binding pocket 903:935 a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes 903:1001 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 7 11 theme folate 1427:1432 arg1 system 1443:1448 the folate receptor system 1423:1448 the folate receptor system 1423:1448 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 2 12 theme low 260:262 arg1 levels 264:269 very low levels 255:269 very low levels in most tissues 255:285 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 7 13 theme extensive 1232:1240 arg1 interactions 1242:1253 The extensive interactions 1228:1253 The extensive interactions between the receptor and ligand 1228:1285 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 5 14 contain has 828:830 arg2 structure 843:851 a globular structure 832:851 a globular structure stabilized by eight disulphide bonds 832:888 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 5 14 contain has 828:830 arg1 FRα 824:826 FRα 824:826 FRα 824:826 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 3 15 theme many 477:480 arg1 tumours 482:488 many tumours 477:488 many tumours 477:488 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 5 16 theme globular 834:841 arg1 structure 843:851 a globular structure 832:851 a globular structure stabilized by eight disulphide bonds 832:888 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 4 17 from resolution 812:821 arg1 acid 798:801 folic acid 792:801 folic acid at 2.8 Å resolution 792:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 4 18 theme 2.8 Å 806:810 arg1 resolution 812:821 2.8 Å resolution 806:821 2.8 Å resolution 806:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 3 19 theme tumours 482:488 arg1 dependency 463:472 The folate dependency 452:472 The folate dependency of many tumours 452:488 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 7 20 theme high 1307:1310 arg1 affinity 1327:1334 the high folate-binding affinity 1303:1334 the high folate-binding affinity of folate receptors 1303:1354 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 7 21 theme receptor 1434:1441 arg1 system 1443:1448 the folate receptor system 1423:1448 the folate receptor system 1423:1448 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 6 22 theme FRα 1215:1217 arg1 binding 1219:1225 FRα binding 1215:1225 FRα binding 1215:1225 The folate pteroate moiety is buried inside the receptor, whereas its glutamate moiety is solvent-exposed and sticks out of the pocket entrance, allowing it to be conjugated to drugs without adversely affecting FRα binding. 23851396 1 23 theme Folate 78:83 arg1 receptors 85:93 Folate receptors 78:93 Folate receptors (FRα, FRβ and FRγ) 78:112 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 23 theme Folate 78:83 arg1 glycoproteins 145:157 cysteine-rich cell-surface glycoproteins 118:157 cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate 118:230 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 23 theme Folate 78:83 arg1 FRγ 109:111 FRγ 109:111 FRγ 109:111 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 23 theme Folate 78:83 arg1 FRβ 101:103 FRβ 101:103 FRβ 101:103 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 23 theme Folate 78:83 arg1 FRα 96:98 FRα 96:98 FRα 96:98 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 2 24 theme high 339:342 arg1 levels 344:349 high levels 339:349 high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions 339:449 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 0 25 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for molecular recognition of folic acid by folate receptors 0:75 Structural basis for molecular recognition of folic acid by folate receptors. 23851396 5 26 theme open 910:913 arg1 pocket 930:935 a deep open folate-binding pocket 903:935 a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes 903:1001 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 7 27 theme folate 1339:1344 arg1 receptors 1346:1354 folate receptors 1339:1354 folate receptors 1339:1354 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 5 28 theme disulphide 873:882 arg1 bonds 884:888 eight disulphide bonds 867:888 eight disulphide bonds 867:888 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 0 29 theme molecular 21:29 arg1 recognition 31:41 molecular recognition 21:41 molecular recognition of folic acid by folate receptors 21:75 Structural basis for molecular recognition of folic acid by folate receptors. 23851396 7 30 theme folate-binding 1312:1325 arg1 affinity 1327:1334 the high folate-binding affinity 1303:1334 the high folate-binding affinity of folate receptors 1303:1354 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 2 31 theme most 274:277 arg1 tissues 279:285 most tissues 274:285 most tissues 274:285 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 5 32 theme receptor 985:992 arg1 subtypes 994:1001 all receptor subtypes 981:1001 all receptor subtypes 981:1001 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 6 33 dep buried 1034:1039 arg1 whereas 1062:1068 whereas 1062:1068 whereas 1062:1068 The folate pteroate moiety is buried inside the receptor, whereas its glutamate moiety is solvent-exposed and sticks out of the pocket entrance, allowing it to be conjugated to drugs without adversely affecting FRα binding. 23851396 7 34 theme specific 1398:1405 arg1 drugs 1407:1411 more specific drugs 1393:1411 more specific drugs targeting the folate receptor system 1393:1448 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 3 35 theme imaging 626:632 arg1 agents 634:639 folate-based imaging agents 613:639 folate-based imaging agents 613:639 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 1 36 theme cell-surface 132:143 arg1 receptors 85:93 Folate receptors 78:93 Folate receptors (FRα, FRβ and FRγ) 78:112 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 36 theme cell-surface 132:143 arg1 glycoproteins 145:157 cysteine-rich cell-surface glycoproteins 118:157 cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate 118:230 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 1 37 theme high 181:184 arg1 affinity 186:193 high affinity 181:193 high affinity 181:193 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 3 38 theme agents 634:639 arg1 administration 547:560 administration 547:560 administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins 547:678 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 4 39 theme folic 792:796 arg1 acid 798:801 folic acid 792:801 folic acid at 2.8 Å resolution 792:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 23851396 6 40 theme pteroate 1015:1022 arg1 moiety 1024:1029 The folate pteroate moiety 1004:1029 The folate pteroate moiety 1004:1029 The folate pteroate moiety is buried inside the receptor, whereas its glutamate moiety is solvent-exposed and sticks out of the pocket entrance, allowing it to be conjugated to drugs without adversely affecting FRα binding. 23851396 2 41 theme cells 417:421 arg1 demand 390:395 the folate demand 379:395 the folate demand of rapidly dividing cells 379:421 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 2 42 theme folate 383:388 arg1 demand 390:395 the folate demand 379:395 the folate demand of rapidly dividing cells 379:421 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 2 43 theme numerous 354:361 arg1 cancers 363:369 numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions 354:449 numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions 354:449 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 0 44 theme acid 52:55 arg1 recognition 31:41 molecular recognition 21:41 molecular recognition of folic acid by folate receptors 21:75 Structural basis for molecular recognition of folic acid by folate receptors. 23851396 6 45 theme folate 1008:1013 arg1 moiety 1024:1029 The folate pteroate moiety 1004:1029 The folate pteroate moiety 1004:1029 The folate pteroate moiety is buried inside the receptor, whereas its glutamate moiety is solvent-exposed and sticks out of the pocket entrance, allowing it to be conjugated to drugs without adversely affecting FRα binding. 23851396 3 46 theme folate-based 613:624 arg1 agents 634:639 folate-based imaging agents 613:639 folate-based imaging agents 613:639 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 7 47 theme receptors 1346:1354 arg1 affinity 1327:1334 the high folate-binding affinity 1303:1334 the high folate-binding affinity of folate receptors 1303:1354 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 5 48 contain contains 894:901 arg1 FRα 824:826 FRα 824:826 FRα 824:826 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 5 48 contain contains 894:901 arg2 pocket 930:935 a deep open folate-binding pocket 903:935 a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes 903:1001 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 0 49 theme folic 46:50 arg1 acid 52:55 folic acid 46:55 folic acid 46:55 Structural basis for molecular recognition of folic acid by folate receptors. 23851396 1 50 with folate 169:174 arg1 affinity 186:193 high affinity 181:193 high affinity 181:193 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 3 51 theme drugs 663:667 arg1 administration 547:560 administration 547:560 administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins 547:678 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 3 52 theme folate 456:461 arg1 dependency 463:472 The folate dependency 452:472 The folate dependency of many tumours 452:488 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 7 53 dep receptor 1267:1274 arg1 the 1263:1265 the 1263:1265 the 1263:1265 The extensive interactions between the receptor and ligand readily explain the high folate-binding affinity of folate receptors and provide a template for designing more specific drugs targeting the folate receptor system. 23851396 3 54 theme high-affinity 586:598 arg1 antifolates 600:610 high-affinity antifolates 586:610 high-affinity antifolates 586:610 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 3 55 theme anti-FRα 565:572 arg1 antibodies 574:583 anti-FRα antibodies 565:583 anti-FRα antibodies 565:583 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 2 56 from levels 344:349 arg1 cancers 363:369 numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions 354:449 numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions 354:449 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 5 57 theme folate-binding 915:928 arg1 pocket 930:935 a deep open folate-binding pocket 903:935 a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes 903:1001 FRα has a globular structure stabilized by eight disulphide bonds and contains a deep open folate-binding pocket comprised of residues that are conserved in all receptor subtypes. 23851396 6 58 theme pocket 1132:1137 arg1 entrance 1139:1146 the pocket entrance 1128:1146 the pocket entrance 1128:1146 The folate pteroate moiety is buried inside the receptor, whereas its glutamate moiety is solvent-exposed and sticks out of the pocket entrance, allowing it to be conjugated to drugs without adversely affecting FRα binding. 23851396 3 59 theme antifolates 600:610 arg1 administration 547:560 administration 547:560 administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins 547:678 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 2 60 theme folate 288:293 arg1 receptors 295:303 folate receptors 288:303 folate receptors 288:303 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 2 60 theme folate 288:293 arg1 FRα 317:319 FRα 317:319 especially FRα 306:319 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 2 61 from levels 264:269 arg1 tissues 279:285 most tissues 274:285 most tissues 274:285 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 1 62 theme cellular 206:213 arg1 uptake 215:220 cellular uptake 206:220 cellular uptake of folate 206:230 Folate receptors (FRα, FRβ and FRγ) are cysteine-rich cell-surface glycoproteins that bind folate with high affinity to mediate cellular uptake of folate. 23851396 0 63 theme folate 60:65 arg1 receptors 67:75 folate receptors 60:75 folate receptors 60:75 Structural basis for molecular recognition of folic acid by folate receptors. 23851396 3 64 theme folate-conjugated 645:661 arg1 drugs 663:667 folate-conjugated drugs 645:667 folate-conjugated drugs 645:667 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 3 65 theme antibodies 574:583 arg1 administration 547:560 administration 547:560 administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins 547:678 The folate dependency of many tumours has been therapeutically and diagnostically exploited by administration of anti-FRα antibodies, high-affinity antifolates, folate-based imaging agents and folate-conjugated drugs and toxins. 23851396 6 66 theme glutamate 1074:1082 arg1 moiety 1084:1089 its glutamate moiety 1070:1089 its glutamate moiety 1070:1089 The folate pteroate moiety is buried inside the receptor, whereas its glutamate moiety is solvent-exposed and sticks out of the pocket entrance, allowing it to be conjugated to drugs without adversely affecting FRα binding. 23851396 2 67 theme dividing 408:415 arg1 cells 417:421 rapidly dividing cells 400:421 rapidly dividing cells 400:421 Although expressed at very low levels in most tissues, folate receptors, especially FRα, are expressed at high levels in numerous cancers to meet the folate demand of rapidly dividing cells under low folate conditions. 23851396 4 68 with complex 779:785 arg1 acid 798:801 folic acid 792:801 folic acid at 2.8 Å resolution 792:821 To understand how folate binds its receptors, we determined the crystal structure of human FRα in complex with folic acid at 2.8 Å resolution. 16492764 2 0 theme GCSF 449:452 arg1 domains 427:433 the Ig-like domains 415:433 the Ig-like domains of GCSF-R and GCSF 415:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 2 0 theme GCSF 449:452 arg1 GCSF 449:452 GCSF 449:452 GCSF 449:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 2 0 theme GCSF 449:452 arg1 GCSF-R 438:443 GCSF-R 438:443 GCSF-R 438:443 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 1 1 theme complex 156:162 arg1 structure 129:137 A crystal structure 119:137 A crystal structure 119:137 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 2 theme GCSF 85:88 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 3 3 theme IL-6/gp130 623:632 arg1 complex 644:650 the IL-6/gp130 signaling complex 619:650 the IL-6/gp130 signaling complex 619:650 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 2 4 theme GCSF 323:326 arg1 complex 335:341 The GCSF:GCSF-R complex 319:341 The GCSF:GCSF-R complex 319:341 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 1 5 theme GCSF 254:257 arg1 GCSF-R 269:274 GCSF-R 269:274 GCSF-R 269:274 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 5 theme GCSF 254:257 arg1 receptor 259:266 GCSF receptor 254:266 GCSF receptor (GCSF-R) 254:275 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 6 theme signaling 100:108 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 3 7 theme signaling 634:642 arg1 complex 644:650 the IL-6/gp130 signaling complex 619:650 the IL-6/gp130 signaling complex 619:650 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 1 8 theme receptor 259:266 arg1 GCSF 217:220 GCSF 217:220 GCSF 217:220 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 8 theme receptor 259:266 arg1 factor 209:214 human granulocyte colony-stimulating factor 172:214 human granulocyte colony-stimulating factor (GCSF) 172:221 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 8 theme receptor 259:266 arg1 region 244:249 a ligand binding region 227:249 a ligand binding region of GCSF receptor (GCSF-R) 227:275 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 9 theme receptor 91:98 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 4 10 theme domain 665:670 arg1 structure 683:691 The Ig-like domain cross-over structure 653:691 The Ig-like domain cross-over structure necessary for GCSF-R activation 653:723 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 10 theme domain 665:670 arg1 consistent 728:737 consistent 728:737 consistent 728:737 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 11 theme necessary 693:701 arg1 structure 683:691 The Ig-like domain cross-over structure 653:691 The Ig-like domain cross-over structure necessary for GCSF-R activation 653:723 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 11 theme necessary 693:701 arg1 consistent 728:737 consistent 728:737 consistent 728:737 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 1 12 theme human 172:176 arg1 GCSF 217:220 GCSF 217:220 GCSF 217:220 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 12 theme human 172:176 arg1 factor 209:214 human granulocyte colony-stimulating factor 172:214 human granulocyte colony-stimulating factor (GCSF) 172:221 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 13 theme complex 110:116 arg1 structure 23:31 Homodimeric cross-over structure 0:31 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 0:117 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 2 14 theme GCSF-R 438:443 arg1 domains 427:433 the Ig-like domains 415:433 the Ig-like domains of GCSF-R and GCSF 415:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 2 14 theme GCSF-R 438:443 arg1 GCSF 449:452 GCSF 449:452 GCSF 449:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 2 14 theme GCSF-R 438:443 arg1 GCSF-R 438:443 GCSF-R 438:443 GCSF-R 438:443 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 1 15 theme granulocyte 178:188 arg1 GCSF 217:220 GCSF 217:220 GCSF 217:220 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 15 theme granulocyte 178:188 arg1 factor 209:214 human granulocyte colony-stimulating factor 172:214 human granulocyte colony-stimulating factor (GCSF) 172:221 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 16 theme cross-over 12:21 arg1 structure 23:31 Homodimeric cross-over structure 0:31 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 0:117 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 3 17 theme complex 479:485 arg1 different 496:504 different 496:504 different 496:504 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 17 theme complex 479:485 arg1 conformation 459:470 The conformation 455:470 The conformation of the complex 455:485 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 18 theme human 524:528 arg1 GCSF 530:533 human GCSF 524:533 human GCSF 524:533 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 1 19 theme colony-stimulating 190:207 arg1 GCSF 217:220 GCSF 217:220 GCSF 217:220 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 19 theme colony-stimulating 190:207 arg1 factor 209:214 human granulocyte colony-stimulating factor 172:214 human granulocyte colony-stimulating factor (GCSF) 172:221 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 20 theme Homodimeric 0:10 arg1 structure 23:31 Homodimeric cross-over structure 0:31 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 0:117 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 4 21 with consistent 728:737 arg1 analyses 793:800 previously reported thermodynamic and mutational analyses 744:800 previously reported thermodynamic and mutational analyses 744:800 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 22 theme GCSF-R 707:712 arg1 activation 714:723 GCSF-R activation 707:723 GCSF-R activation 707:723 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 2 23 theme cross-over 384:393 arg1 interaction 395:405 a cross-over interaction 382:405 a cross-over interaction between the Ig-like domains of GCSF-R and GCSF 382:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 0 24 theme human 40:44 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 3 25 theme cytokine 543:550 arg1 domain 572:577 the cytokine receptor homologous domain 539:577 the cytokine receptor homologous domain of mouse GCSF-R 539:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 25 theme cytokine 543:550 arg1 GCSF-R 588:593 mouse GCSF-R 582:593 mouse GCSF-R 582:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 26 theme GCSF-R 588:593 arg1 domain 572:577 the cytokine receptor homologous domain 539:577 the cytokine receptor homologous domain of mouse GCSF-R 539:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 26 theme GCSF-R 588:593 arg1 GCSF-R 588:593 mouse GCSF-R 582:593 mouse GCSF-R 582:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 26 theme GCSF-R 588:593 arg1 GCSF 530:533 human GCSF 524:533 human GCSF 524:533 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 27 theme receptor 552:559 arg1 domain 572:577 the cytokine receptor homologous domain 539:577 the cytokine receptor homologous domain of mouse GCSF-R 539:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 27 theme receptor 552:559 arg1 GCSF-R 588:593 mouse GCSF-R 582:593 mouse GCSF-R 582:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 0 28 theme colony-stimulating 58:75 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 4 29 theme Ig-like 657:663 arg1 structure 683:691 The Ig-like domain cross-over structure 653:691 The Ig-like domain cross-over structure necessary for GCSF-R activation 653:723 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 29 theme Ig-like 657:663 arg1 consistent 728:737 consistent 728:737 consistent 728:737 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 0 30 theme granulocyte 46:56 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 2 31 theme Ig-like 419:425 arg1 domains 427:433 the Ig-like domains 415:433 the Ig-like domains of GCSF-R and GCSF 415:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 2 31 theme Ig-like 419:425 arg1 GCSF 449:452 GCSF 449:452 GCSF 449:452 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 2 31 theme Ig-like 419:425 arg1 GCSF-R 438:443 GCSF-R 438:443 GCSF-R 438:443 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 1 32 theme crystal 121:127 arg1 structure 129:137 A crystal structure 119:137 A crystal structure 119:137 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 3 33 from that 511:514 arg1 different 496:504 different 496:504 different 496:504 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 33 from that 511:514 arg1 conformation 459:470 The conformation 455:470 The conformation of the complex 455:485 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 4 34 theme reported 755:762 arg1 analyses 793:800 previously reported thermodynamic and mutational analyses 744:800 previously reported thermodynamic and mutational analyses 744:800 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 35 theme mutational 782:791 arg1 analyses 793:800 previously reported thermodynamic and mutational analyses 744:800 previously reported thermodynamic and mutational analyses 744:800 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 36 theme cross-over 672:681 arg1 structure 683:691 The Ig-like domain cross-over structure 653:691 The Ig-like domain cross-over structure necessary for GCSF-R activation 653:723 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 4 36 theme cross-over 672:681 arg1 consistent 728:737 consistent 728:737 consistent 728:737 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 3 37 theme mouse 582:586 arg1 GCSF-R 588:593 mouse GCSF-R 582:593 mouse GCSF-R 582:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 1 38 theme signaling 146:154 arg1 complex 156:162 the signaling complex 142:162 the signaling complex 142:162 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 1 39 theme ligand 229:234 arg1 region 244:249 a ligand binding region 227:249 a ligand binding region of GCSF receptor (GCSF-R) 227:275 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 3 40 theme homologous 561:570 arg1 domain 572:577 the cytokine receptor homologous domain 539:577 the cytokine receptor homologous domain of mouse GCSF-R 539:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 3 40 theme homologous 561:570 arg1 GCSF-R 588:593 mouse GCSF-R 582:593 mouse GCSF-R 582:593 The conformation of the complex is quite different from that between human GCSF and the cytokine receptor homologous domain of mouse GCSF-R, but similar to that of the IL-6/gp130 signaling complex. 16492764 2 41 theme 2:2 352:354 arg1 stoichiometry 356:368 a 2:2 stoichiometry 350:368 a 2:2 stoichiometry 350:368 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 16492764 1 42 theme binding 236:242 arg1 region 244:249 a ligand binding region 227:249 a ligand binding region of GCSF receptor (GCSF-R) 227:275 A crystal structure of the signaling complex between human granulocyte colony-stimulating factor (GCSF) and a ligand binding region of GCSF receptor (GCSF-R), has been determined to 2.8 A resolution. 16492764 0 43 theme factor 77:82 arg1 complex 110:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex 36:116 Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. 16492764 4 44 theme thermodynamic 764:776 arg1 analyses 793:800 previously reported thermodynamic and mutational analyses 744:800 previously reported thermodynamic and mutational analyses 744:800 The Ig-like domain cross-over structure necessary for GCSF-R activation is consistent with previously reported thermodynamic and mutational analyses. 16492764 2 45 theme GCSF-R 328:333 arg1 complex 335:341 The GCSF:GCSF-R complex 319:341 The GCSF:GCSF-R complex 319:341 The GCSF:GCSF-R complex formed a 2:2 stoichiometry by means of a cross-over interaction between the Ig-like domains of GCSF-R and GCSF. 18042673 3 0 theme unliganded 455:464 arg1 enzymes 486:492 the unliganded and inhibitor-bound enzymes 451:492 the unliganded and inhibitor-bound enzymes 451:492 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 1 1 theme responsible 147:157 arg1 Aggrecanases 85:96 Aggrecanases 85:96 Aggrecanases 85:96 Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. 18042673 1 1 theme responsible 147:157 arg1 proteinases 135:145 the principal proteinases 121:145 the principal proteinases responsible for aggrecan degradation in osteoarthritis 121:200 Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. 18042673 3 2 dep configurations 543:556 arg1 form 596:599 an autoinhibited, nonbinding, closed form 559:599 an autoinhibited, nonbinding, closed form 559:599 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 2 dep configurations 543:556 arg1 form 622:625 an open, binding form 605:625 an open, binding form 605:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 2 dep configurations 543:556 arg1 configurations 543:556 two essentially different catalytic-site configurations 502:556 two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form 502:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 3 theme autoinhibited 562:574 arg1 form 596:599 an autoinhibited, nonbinding, closed form 559:599 an autoinhibited, nonbinding, closed form 559:599 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 3 theme autoinhibited 562:574 arg1 configurations 543:556 two essentially different catalytic-site configurations 502:556 two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form 502:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 4 4 theme ensemble 691:698 arg1 one 730:732 one 730:732 one 730:732 On this basis, we propose that mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active. 18042673 4 4 theme ensemble 691:698 arg1 ensemble 691:698 an ensemble 688:698 an ensemble 688:698 On this basis, we propose that mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active. 18042673 3 5 theme different 518:526 arg1 configurations 543:556 two essentially different catalytic-site configurations 502:556 two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form 502:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 5 theme different 518:526 arg1 form 622:625 an open, binding form 605:625 an open, binding form 605:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 5 theme different 518:526 arg1 form 596:599 an autoinhibited, nonbinding, closed form 559:599 an autoinhibited, nonbinding, closed form 559:599 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 2 6 dep enzyme 401:406 arg1 form 418:421 apo form 414:421 apo form 414:421 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 7 theme bound 365:369 arg1 inhibitor 371:379 bound inhibitor 365:379 bound inhibitor 365:379 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 0 8 theme aggrecan 36:43 arg1 enzymes 55:61 the two major aggrecan degrading enzymes 22:61 the two major aggrecan degrading enzymes 22:61 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 2 9 theme crystal 253:259 arg1 structures 261:270 crystal structures 253:270 crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5 253:341 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 9 theme crystal 253:259 arg1 each 344:347 each 344:347 each 344:347 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 1 10 theme aggrecan 163:170 arg1 degradation 172:182 aggrecan degradation 163:182 aggrecan degradation in osteoarthritis 163:200 Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. 18042673 2 11 theme human 295:299 arg1 ADAMTS5 335:341 ADAMTS5 335:341 ADAMTS5 335:341 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 11 theme human 295:299 arg1 ADAMTS4 323:329 ADAMTS4 323:329 ADAMTS4 323:329 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 11 theme human 295:299 arg1 isoforms 313:320 the two most active human aggrecanase isoforms 275:320 the two most active human aggrecanase isoforms 275:320 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 0 12 theme Crystal 0:6 arg1 ADAMTS4 64:70 ADAMTS4 64:70 ADAMTS4 64:70 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 0 12 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of the two major aggrecan degrading enzymes 0:61 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 0 12 theme Crystal 0:6 arg1 ADAMTS5 76:82 ADAMTS5 76:82 ADAMTS5 76:82 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 2 13 theme active 288:293 arg1 ADAMTS5 335:341 ADAMTS5 335:341 ADAMTS5 335:341 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 13 theme active 288:293 arg1 ADAMTS4 323:329 ADAMTS4 323:329 ADAMTS4 323:329 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 13 theme active 288:293 arg1 isoforms 313:320 the two most active human aggrecanase isoforms 275:320 the two most active human aggrecanase isoforms 275:320 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 3 14 theme catalytic-site 528:541 arg1 configurations 543:556 two essentially different catalytic-site configurations 502:556 two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form 502:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 14 theme catalytic-site 528:541 arg1 form 622:625 an open, binding form 605:625 an open, binding form 605:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 14 theme catalytic-site 528:541 arg1 form 596:599 an autoinhibited, nonbinding, closed form 559:599 an autoinhibited, nonbinding, closed form 559:599 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 0 15 theme major 30:34 arg1 enzymes 55:61 the two major aggrecan degrading enzymes 22:61 the two major aggrecan degrading enzymes 22:61 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 2 16 theme drug 230:233 arg1 target 235:240 a drug target 228:240 a drug target 228:240 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 3 17 dep autoinhibited 562:574 arg1 nonbinding 577:586 nonbinding 577:586 nonbinding 577:586 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 17 dep autoinhibited 562:574 arg1 closed 589:594 closed 589:594 closed 589:594 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 18 theme open 608:611 arg1 configurations 543:556 two essentially different catalytic-site configurations 502:556 two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form 502:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 3 18 theme open 608:611 arg1 form 622:625 an open, binding form 605:625 an open, binding form 605:625 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 0 19 theme degrading 45:53 arg1 enzymes 55:61 the two major aggrecan degrading enzymes 22:61 the two major aggrecan degrading enzymes 22:61 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 2 20 dep wherein 389:395 arg1 enzyme 401:406 the enzyme 397:406 one wherein the enzyme is in apo form 385:421 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 21 theme apo 414:416 arg1 form 418:421 apo form 414:421 apo form 414:421 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 4 22 theme mature 659:664 arg1 aggrecanases 666:677 mature aggrecanases 659:677 mature aggrecanases 659:677 On this basis, we propose that mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active. 18042673 2 23 theme isoforms 313:320 arg1 structures 261:270 crystal structures 253:270 crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5 253:341 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 23 theme isoforms 313:320 arg1 each 344:347 each 344:347 each 344:347 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 3 24 dep open 608:611 arg1 binding 614:620 binding 614:620 binding 614:620 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 4 25 theme isomers 716:722 arg1 ensemble 691:698 an ensemble 688:698 an ensemble 688:698 On this basis, we propose that mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active. 18042673 1 26 from degradation 172:182 arg1 osteoarthritis 187:200 osteoarthritis 187:200 osteoarthritis 187:200 Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. 18042673 2 27 theme aggrecanase 301:311 arg1 ADAMTS5 335:341 ADAMTS5 335:341 ADAMTS5 335:341 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 27 theme aggrecanase 301:311 arg1 ADAMTS4 323:329 ADAMTS4 323:329 ADAMTS4 323:329 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 27 theme aggrecanase 301:311 arg1 isoforms 313:320 the two most active human aggrecanase isoforms 275:320 the two most active human aggrecanase isoforms 275:320 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 3 28 theme inhibitor-bound 470:484 arg1 enzymes 486:492 the unliganded and inhibitor-bound enzymes 451:492 the unliganded and inhibitor-bound enzymes 451:492 These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. 18042673 0 29 theme enzymes 55:61 arg1 ADAMTS4 64:70 ADAMTS4 64:70 ADAMTS4 64:70 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 0 29 theme enzymes 55:61 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of the two major aggrecan degrading enzymes 0:61 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 0 29 theme enzymes 55:61 arg1 ADAMTS5 76:82 ADAMTS5 76:82 ADAMTS5 76:82 Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5. 18042673 2 30 with complex 352:358 arg1 inhibitor 371:379 bound inhibitor 365:379 bound inhibitor 365:379 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 2 30 with complex 352:358 arg1 wherein 389:395 one wherein 385:395 one wherein the enzyme is in apo form 385:421 Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. 18042673 1 31 theme principal 125:133 arg1 Aggrecanases 85:96 Aggrecanases 85:96 Aggrecanases 85:96 Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. 18042673 1 31 theme principal 125:133 arg1 proteinases 135:145 the principal proteinases 121:145 the principal proteinases responsible for aggrecan degradation in osteoarthritis 121:200 Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. 26902720 6 0 theme Wnt5a 887:891 arg1 subtypes 918:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 6 1 theme afamin 928:933 arg1 complexation 935:946 afamin complexation 928:946 afamin complexation 928:946 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 0 2 theme glycoprotein 80:91 arg1 afamin/α-albumin 93:108 a serum glycoprotein afamin/α-albumin 72:108 a serum glycoprotein afamin/α-albumin 72:108 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 2 3 from use 303:305 arg1 assays 339:344 various in vitro and in vivo assays 310:344 various in vitro and in vivo assays 310:344 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 26902720 4 4 theme growth 732:737 arg1 assay 739:743 the intestical stem cell growth assay 707:743 the intestical stem cell growth assay 707:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 4 5 theme intestical 711:720 arg1 assay 739:743 the intestical stem cell growth assay 707:743 the intestical stem cell growth assay 707:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 0 6 theme serum 74:78 arg1 afamin/α-albumin 93:108 a serum glycoprotein afamin/α-albumin 72:108 a serum glycoprotein afamin/α-albumin 72:108 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 4 7 theme cell 727:730 arg1 assay 739:743 the intestical stem cell growth assay 707:743 the intestical stem cell growth assay 707:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 4 8 theme 1:1 562:564 arg1 complex 566:572 a 1:1 complex 560:572 a 1:1 complex 560:572 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 4 9 theme various 672:678 arg1 assay 739:743 the intestical stem cell growth assay 707:743 the intestical stem cell growth assay 707:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 4 9 theme various 672:678 arg1 systems 689:695 various cellular systems 672:695 various cellular systems including the intestical stem cell growth assay 672:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 6 10 theme thorough 1030:1037 arg1 scrutiny 1051:1058 a thorough biochemical scrutiny 1028:1058 a thorough biochemical scrutiny that had been difficult for years 1028:1092 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 6 10 theme thorough 1030:1037 arg1 difficult 1074:1082 difficult 1074:1082 difficult 1074:1082 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 3 11 theme bovine 512:517 arg1 serum 519:523 the bovine serum 508:523 the bovine serum included in the media 508:545 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 6 12 theme signaling 1001:1009 arg1 mechanisms 1011:1020 their signaling mechanisms 995:1020 their signaling mechanisms 995:1020 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 1 13 theme various 162:168 arg1 diseases 170:177 various diseases 162:177 various diseases 162:177 Wnt plays important role during development and in various diseases. 26902720 3 14 theme tagged 372:377 arg1 Wnt3a 391:395 N-terminally tagged recombinant Wnt3a 359:395 N-terminally tagged recombinant Wnt3a secreted from cells 359:415 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 3 15 theme recombinant 379:389 arg1 Wnt3a 391:395 N-terminally tagged recombinant Wnt3a 359:395 N-terminally tagged recombinant Wnt3a secreted from cells 359:415 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 2 16 dep in 331:332 arg1 vivo 334:337 vivo 334:337 vivo 334:337 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 26902720 6 17 theme more 909:912 arg1 subtypes 918:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 3 18 gly glycoprotein 475:486 arg1 glycoprotein 475:486 a glycoprotein afamin 473:493 a glycoprotein afamin derived from the bovine serum included in the media 473:545 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 4 19 from buffer 620:625 arg1 soluble 601:607 soluble 601:607 soluble 601:607 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 5 20 theme large 841:845 arg1 quantity 847:854 large quantity 841:854 large quantity 841:854 By co-expressing with afamin, biologically active afamin-Wnt complex can be easily obtained in large quantity. 26902720 0 21 theme Active 0:5 arg1 form 25:28 Active and water-soluble form 0:28 Active and water-soluble form of lipidated Wnt protein 0:53 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 0 22 gly glycoprotein 80:91 arg1 glycoprotein 80:91 a serum glycoprotein afamin/α-albumin 72:108 a serum glycoprotein afamin/α-albumin 72:108 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 5 23 theme active 789:794 arg1 complex 807:813 biologically active afamin-Wnt complex 776:813 biologically active afamin-Wnt complex 776:813 By co-expressing with afamin, biologically active afamin-Wnt complex can be easily obtained in large quantity. 26902720 2 24 dep in 318:319 arg1 vitro 321:325 vitro 321:325 vitro 321:325 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 26902720 0 25 theme water-soluble 11:23 arg1 form 25:28 Active and water-soluble form 0:28 Active and water-soluble form of lipidated Wnt protein 0:53 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 6 26 theme Wnt 914:916 arg1 subtypes 918:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 2 27 theme detergents 276:285 arg1 presence 264:271 the presence 260:271 the presence of detergents 260:285 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 26902720 4 28 from soluble 601:607 arg1 buffer 620:625 aqueous buffer 612:625 aqueous buffer 612:625 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 0 29 theme lipidated 33:41 arg1 protein 47:53 lipidated Wnt protein 33:53 lipidated Wnt protein 33:53 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 3 30 theme glycoprotein 475:486 arg1 afamin 488:493 a glycoprotein afamin 473:493 a glycoprotein afamin derived from the bovine serum included in the media 473:545 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 6 31 theme Wnt 979:981 arg1 ligands 983:989 various Wnt ligands 971:989 various Wnt ligands 971:989 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 4 32 theme aqueous 612:618 arg1 buffer 620:625 aqueous buffer 612:625 aqueous buffer 612:625 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 6 33 theme many 904:907 arg1 subtypes 918:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 6 34 theme various 971:977 arg1 ligands 983:989 various Wnt ligands 971:989 various Wnt ligands 971:989 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 0 35 theme protein 47:53 arg1 form 25:28 Active and water-soluble form 0:28 Active and water-soluble form of lipidated Wnt protein 0:53 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 2 36 theme in 318:319 arg1 assays 339:344 various in vitro and in vivo assays 310:344 various in vitro and in vivo assays 310:344 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 26902720 0 37 theme Wnt 43:45 arg1 protein 47:53 lipidated Wnt protein 33:53 lipidated Wnt protein 33:53 Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin. 26902720 3 38 attach derived 495:501 arg2 afamin 488:493 a glycoprotein afamin 473:493 a glycoprotein afamin derived from the bovine serum included in the media 473:545 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 3 38 attach derived 495:501 arg1 serum 519:523 the bovine serum 508:523 the bovine serum included in the media 508:545 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 2 39 theme various 310:316 arg1 assays 339:344 various in vitro and in vivo assays 310:344 various in vitro and in vivo assays 310:344 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 26902720 6 40 theme Wnt3 894:897 arg1 subtypes 918:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 Wnt5a, Wnt3, and many more Wnt subtypes 887:925 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 4 41 with complex 566:572 arg1 afamin 579:584 afamin 579:584 afamin 579:584 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 1 42 theme important 121:129 arg1 role 131:134 important role 121:134 important role 121:134 Wnt plays important role during development and in various diseases. 26902720 4 43 theme stem 722:725 arg1 assay 739:743 the intestical stem cell growth assay 707:743 the intestical stem cell growth assay 707:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 4 44 theme cellular 680:687 arg1 assay 739:743 the intestical stem cell growth assay 707:743 the intestical stem cell growth assay 707:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 4 44 theme cellular 680:687 arg1 systems 689:695 various cellular systems 672:695 various cellular systems including the intestical stem cell growth assay 672:743 Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestical stem cell growth assay. 26902720 6 45 theme biochemical 1039:1049 arg1 scrutiny 1051:1058 a thorough biochemical scrutiny 1028:1058 a thorough biochemical scrutiny that had been difficult for years 1028:1092 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 6 45 theme biochemical 1039:1049 arg1 difficult 1074:1082 difficult 1074:1082 difficult 1074:1082 As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, afamin complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years. 26902720 3 46 theme N-terminally 359:370 arg1 Wnt3a 391:395 N-terminally tagged recombinant Wnt3a 359:395 N-terminally tagged recombinant Wnt3a secreted from cells 359:415 We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. 26902720 5 47 theme afamin-Wnt 796:805 arg1 complex 807:813 biologically active afamin-Wnt complex 776:813 biologically active afamin-Wnt complex 776:813 By co-expressing with afamin, biologically active afamin-Wnt complex can be easily obtained in large quantity. 26902720 2 48 theme in 331:332 arg1 assays 339:344 various in vitro and in vivo assays 310:344 various in vitro and in vivo assays 310:344 Because Wnts are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. 24374484 0 0 theme drug 103:106 arg1 design 108:113 drug design 103:113 drug design of selective inhibitors 103:137 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 1 1 from cord 339:342 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 2 from kidneys 271:277 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 3 theme CA 172:173 arg1 XIV 167:169 Carbonic anhydrase isoform XIV 140:169 Carbonic anhydrase isoform XIV (CA XIV) 140:178 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 3 theme CA 172:173 arg1 XIV 175:177 CA XIV 172:177 CA XIV 172:177 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 4 theme family 219:224 arg1 XIV 167:169 Carbonic anhydrase isoform XIV 140:169 Carbonic anhydrase isoform XIV (CA XIV) 140:178 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 4 theme family 219:224 arg1 member 192:197 the last member 183:197 the last member of the human (h) CA family discovered so far 183:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 5 theme small 287:291 arg1 intestine 293:301 small intestine 287:301 small intestine 287:301 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 4 6 theme entire 757:762 arg1 domain 778:783 the entire extracellular domain 753:783 the entire extracellular domain of this enzyme 753:798 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 4 6 theme entire 757:762 arg1 enzyme 793:798 this enzyme 788:798 this enzyme 788:798 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 4 7 theme extracellular 764:776 arg1 domain 778:783 the entire extracellular domain 753:783 the entire extracellular domain of this enzyme 753:798 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 4 7 theme extracellular 764:776 arg1 enzyme 793:798 this enzyme 788:798 this enzyme 788:798 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 3 8 theme putative 597:604 arg1 region 620:625 a putative transmembrane region 595:625 a putative transmembrane region 595:625 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 1 9 from brain 264:268 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 5 10 theme α-CA 852:855 arg1 fold 857:860 the typical α-CA fold 840:860 the typical α-CA fold 840:860 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 11 theme oligomeric 1050:1059 arg1 arrangement 1061:1071 the diverse oligomeric arrangement 1038:1071 the diverse oligomeric arrangement 1038:1071 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 12 theme other 960:964 arg1 membrane 966:973 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 4 13 theme domain 778:783 arg1 expression 689:698 expression 689:698 expression 689:698 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 4 13 theme domain 778:783 arg1 structure 740:748 the crystallographic structure 719:748 the crystallographic structure 719:748 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 4 13 theme domain 778:783 arg1 purification 701:712 purification 701:712 purification 701:712 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 5 14 theme structure 821:829 arg1 analysis 805:812 The analysis 801:812 The analysis of the structure 801:829 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 0 15 theme inhibitors 128:137 arg1 design 108:113 drug design 103:113 drug design of selective inhibitors 103:137 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 1 16 theme urinary 304:310 arg1 bladder 312:318 urinary bladder 304:318 urinary bladder 304:318 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 kidneys 271:277 kidneys 271:277 kidneys 271:277 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 bladder 312:318 urinary bladder 304:318 urinary bladder 304:318 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 cord 339:342 spinal cord 332:342 spinal cord 332:342 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 colon 280:284 colon 280:284 colon 280:284 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 liver 321:325 liver 321:325 liver 321:325 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 intestine 293:301 small intestine 287:301 small intestine 287:301 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 17 from localized 251:259 arg1 brain 264:268 brain 264:268 brain 264:268 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 5 18 theme 10-stranded 874:884 arg1 β-sheet 886:892 a 10-stranded β-sheet 872:892 a 10-stranded β-sheet 872:892 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 0 19 theme selective 118:126 arg1 inhibitors 128:137 selective inhibitors 118:137 selective inhibitors 118:137 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 5 20 theme associated 975:984 arg1 XII 1013:1015 XII 1013:1015 XII 1013:1015 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 20 theme associated 975:984 arg1 hCAs 996:999 hCAs IV 996:1002 hCAs IV 996:1002 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 20 theme associated 975:984 arg1 IX 1005:1006 IX 1005:1006 IX 1005:1006 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 20 theme associated 975:984 arg1 isoforms 986:993 associated isoforms 975:993 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 4 21 theme crystallographic 723:738 arg1 structure 740:748 the crystallographic structure 719:748 the crystallographic structure 719:748 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 5 22 theme membrane 1238:1245 arg1 one 1227:1229 one 1227:1229 one 1227:1229 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 22 theme membrane 1238:1245 arg1 membrane 1238:1245 the membrane 1234:1245 the membrane associated α-CAs 1234:1262 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 1 23 from liver 321:325 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 0 24 theme structural 4:13 arg1 comparison 15:24 The structural comparison 0:24 The structural comparison between membrane-associated human carbonic anhydrases 0:78 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 3 25 theme Human 477:481 arg1 hCA 503:505 hCA 503:505 hCA 503:505 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 3 25 theme Human 477:481 arg1 anhydrase 492:500 Human carbonic anhydrase 477:500 Human carbonic anhydrase (hCA) XIV 477:510 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 3 26 theme small 634:638 arg1 tail 652:655 a small cytoplasmic tail 632:655 a small cytoplasmic tail 632:655 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 5 27 located observed 1117:1124 arg1 127-136 1140:1146 the region 127-136 1129:1146 the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs 1129:1262 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 27 located observed 1117:1124 arg2 differences 1105:1115 the sequence and structural differences 1077:1115 differences 1105:1115 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 3 28 theme carbonic 483:490 arg1 hCA 503:505 hCA 503:505 hCA 503:505 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 3 28 theme carbonic 483:490 arg1 anhydrase 492:500 Human carbonic anhydrase 477:500 Human carbonic anhydrase (hCA) XIV 477:510 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 1 29 theme last 187:190 arg1 XIV 167:169 Carbonic anhydrase isoform XIV 140:169 Carbonic anhydrase isoform XIV (CA XIV) 140:178 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 29 theme last 187:190 arg1 member 192:197 the last member 183:197 the last member of the human (h) CA family discovered so far 183:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 2 30 theme skin 464:467 arg1 tumors 469:474 some skin tumors 459:474 some retinopathies as well as some skin tumors 429:474 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 30 theme skin 464:467 arg1 target 395:400 a possible drug target 379:400 a possible drug target for treatment of epilepsy 379:426 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 5 31 theme typical 844:850 arg1 fold 857:860 the typical α-CA fold 840:860 the typical α-CA fold 840:860 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 32 theme main 1155:1158 arg1 factors 1160:1166 the main factors 1151:1166 the main factors 1151:1166 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 3 33 theme cytoplasmic 640:650 arg1 tail 652:655 a small cytoplasmic tail 632:655 a small cytoplasmic tail 632:655 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 2 34 theme drug 390:393 arg1 It 345:346 It 345:346 It 345:346 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 34 theme drug 390:393 arg1 retinopathies 434:446 some retinopathies 429:446 some retinopathies as well as some skin tumors 429:474 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 34 theme drug 390:393 arg1 tumors 469:474 some skin tumors 459:474 some retinopathies as well as some skin tumors 429:474 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 34 theme drug 390:393 arg1 target 395:400 a possible drug target 379:400 a possible drug target for treatment of epilepsy 379:426 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 5 35 with comparison 936:945 arg1 membrane 966:973 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 0 36 theme human 54:58 arg1 anhydrases 69:78 membrane-associated human carbonic anhydrases 34:78 membrane-associated human carbonic anhydrases 34:78 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 5 37 theme diverse 1042:1048 arg1 arrangement 1061:1071 the diverse oligomeric arrangement 1038:1071 the diverse oligomeric arrangement 1038:1071 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 1 38 from intestine 293:301 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 0 39 theme membrane-associated 34:52 arg1 anhydrases 69:78 membrane-associated human carbonic anhydrases 34:78 membrane-associated human carbonic anhydrases 34:78 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 1 40 theme human 206:210 arg1 family 219:224 the human (h) CA family 202:224 the human (h) CA family discovered so far 202:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 41 from colon 280:284 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 5 42 dep isoforms 986:993 arg1 hCAs 996:999 hCAs IV 996:1002 hCAs IV 996:1002 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 42 dep isoforms 986:993 arg1 IX 1005:1006 IX 1005:1006 IX 1005:1006 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 42 dep isoforms 986:993 arg1 XII 1013:1015 XII 1013:1015 XII 1013:1015 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 42 dep isoforms 986:993 arg1 isoforms 986:993 associated isoforms 975:993 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 4 43 theme enzyme 793:798 arg1 domain 778:783 the entire extracellular domain 753:783 the entire extracellular domain of this enzyme 753:798 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 4 43 theme enzyme 793:798 arg1 enzyme 793:798 this enzyme 788:798 this enzyme 788:798 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 1 44 theme spinal 332:337 arg1 cord 339:342 spinal cord 332:342 spinal cord 332:342 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 3 45 theme anhydrase 492:500 arg1 protein 537:543 a membrane-associated protein 515:543 a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail 515:655 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 3 45 theme anhydrase 492:500 arg1 XIV 508:510 Human carbonic anhydrase (hCA) XIV 477:510 Human carbonic anhydrase (hCA) XIV 477:510 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 0 46 theme carbonic 60:67 arg1 anhydrases 69:78 membrane-associated human carbonic anhydrases 34:78 membrane-associated human carbonic anhydrases 34:78 The structural comparison between membrane-associated human carbonic anhydrases provides insights into drug design of selective inhibitors. 24374484 5 47 theme molecule 916:923 arg1 core 904:907 the core 900:907 the core of the molecule 900:923 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 1 48 theme Carbonic 140:147 arg1 anhydrase 149:157 Carbonic anhydrase 140:157 Carbonic anhydrase isoform XIV (CA XIV) 140:178 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 5 49 dep membrane 966:973 arg1 XII 1013:1015 XII 1013:1015 XII 1013:1015 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 49 dep membrane 966:973 arg1 hCAs 996:999 hCAs IV 996:1002 hCAs IV 996:1002 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 49 dep membrane 966:973 arg1 IX 1005:1006 IX 1005:1006 IX 1005:1006 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 5 49 dep membrane 966:973 arg1 isoforms 986:993 associated isoforms 975:993 all the other membrane associated isoforms (hCAs IV, IX, and XII) 952:1016 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 1 50 theme anhydrase 149:157 arg1 member 192:197 the last member 183:197 the last member of the human (h) CA family discovered so far 183:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 50 theme anhydrase 149:157 arg1 XIV 167:169 Carbonic anhydrase isoform XIV 140:169 Carbonic anhydrase isoform XIV (CA XIV) 140:178 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 50 theme anhydrase 149:157 arg1 XIV 175:177 CA XIV 172:177 CA XIV 172:177 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 5 51 theme region 1133:1138 arg1 127-136 1140:1146 the region 127-136 1129:1146 the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs 1129:1262 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 3 52 theme N-terminal 562:571 arg1 domain 587:592 an N-terminal extracellular domain 559:592 an N-terminal extracellular domain 559:592 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 3 53 theme membrane-associated 517:535 arg1 protein 537:543 a membrane-associated protein 515:543 a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail 515:655 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 3 53 theme membrane-associated 517:535 arg1 XIV 508:510 Human carbonic anhydrase (hCA) XIV 477:510 Human carbonic anhydrase (hCA) XIV 477:510 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 1 54 theme isoform 159:165 arg1 member 192:197 the last member 183:197 the last member of the human (h) CA family discovered so far 183:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 54 theme isoform 159:165 arg1 XIV 167:169 Carbonic anhydrase isoform XIV 140:169 Carbonic anhydrase isoform XIV (CA XIV) 140:178 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 54 theme isoform 159:165 arg1 XIV 175:177 CA XIV 172:177 CA XIV 172:177 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 1 55 theme h 213:213 arg1 family 219:224 the human (h) CA family 202:224 the human (h) CA family discovered so far 202:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 2 56 theme epilepsy 419:426 arg1 treatment 406:414 treatment 406:414 treatment of epilepsy 406:426 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 3 57 theme extracellular 573:585 arg1 domain 587:592 an N-terminal extracellular domain 559:592 an N-terminal extracellular domain 559:592 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 5 58 theme selective 1197:1205 arg1 inhibitors 1207:1216 selective inhibitors 1197:1216 selective inhibitors for each one of the membrane associated α-CAs 1197:1262 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 4 59 dep expression 689:698 arg1 the 685:687 the 685:687 the 685:687 In this article, we report the expression, purification, and the crystallographic structure of the entire extracellular domain of this enzyme. 24374484 5 60 theme sequence 1081:1088 arg1 differences 1105:1115 the sequence and structural differences 1077:1115 differences 1105:1115 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 2 61 theme possible 381:388 arg1 It 345:346 It 345:346 It 345:346 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 61 theme possible 381:388 arg1 retinopathies 434:446 some retinopathies 429:446 some retinopathies as well as some skin tumors 429:474 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 61 theme possible 381:388 arg1 tumors 469:474 some skin tumors 459:474 some retinopathies as well as some skin tumors 429:474 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 2 61 theme possible 381:388 arg1 target 395:400 a possible drug target 379:400 a possible drug target for treatment of epilepsy 379:426 It has recently been described as a possible drug target for treatment of epilepsy, some retinopathies as well as some skin tumors. 24374484 3 62 theme transmembrane 606:618 arg1 region 620:625 a putative transmembrane region 595:625 a putative transmembrane region 595:625 Human carbonic anhydrase (hCA) XIV is a membrane-associated protein consisting of an N-terminal extracellular domain, a putative transmembrane region, and a small cytoplasmic tail. 24374484 5 63 theme inhibitors 1207:1216 arg1 design 1187:1192 the design 1183:1192 the design of selective inhibitors for each one of the membrane associated α-CAs 1183:1262 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 1 64 from bladder 312:318 arg1 localized 251:259 localized 251:259 localized 251:259 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 24374484 5 65 theme structural 1094:1103 arg1 differences 1105:1115 the sequence and structural differences 1077:1115 differences 1105:1115 The analysis of the structure revealed the typical α-CA fold, in which a 10-stranded β-sheet forms the core of the molecule, while the comparison with all the other membrane associated isoforms (hCAs IV, IX, and XII) allowed to identify the diverse oligomeric arrangement and the sequence and structural differences observed in the region 127-136 as the main factors to consider in the design of selective inhibitors for each one of the membrane associated α-CAs. 24374484 1 66 theme CA 216:217 arg1 family 219:224 the human (h) CA family 202:224 the human (h) CA family discovered so far 202:242 Carbonic anhydrase isoform XIV (CA XIV) is the last member of the human (h) CA family discovered so far, being localized in brain, kidneys, colon, small intestine, urinary bladder, liver, and spinal cord. 11053835 5 0 from removal 1068:1074 arg1 core 1114:1117 the protein core 1102:1117 the protein core 1102:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 7 1 theme Glu202 1611:1616 arg1 carboxylate 1618:1628 the Glu202 carboxylate 1607:1628 the Glu202 carboxylate 1607:1628 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 5 2 from core 1114:1117 arg1 removal 1068:1074 removal 1068:1074 removal of the charged group from the protein core 1068:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 5 2 from core 1114:1117 arg1 substitution 1127:1138 its substitution 1123:1138 its substitution by a neutral isosteric moiety 1123:1168 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 5 2 from core 1114:1117 arg1 group 1091:1095 the charged group 1079:1095 the charged group from the protein core 1079:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 6 3 theme hydrogen-bond 1297:1309 arg1 network 1311:1317 a hydrogen-bond network 1295:1317 a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme 1295:1488 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 3 theme hydrogen-bond 1297:1309 arg1 One 1237:1239 One 1237:1239 One 1237:1239 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 3 theme hydrogen-bond 1297:1309 arg1 elements 1248:1255 the elements 1244:1255 the elements of this architecture 1244:1276 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 7 4 with interaction 1704:1714 arg1 inhibitors 1760:1769 inhibitors 1760:1769 inhibitors 1760:1769 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 4 with interaction 1704:1714 arg1 substrates 1745:1754 substrates 1745:1754 substrates 1745:1754 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 1 5 theme polypeptide 266:276 arg1 toxin 278:282 a 'three-finger' polypeptide toxin 249:282 a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps) 249:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 1 5 theme polypeptide 266:276 arg1 fasciculin-II 234:246 fasciculin-II 234:246 fasciculin-II 234:246 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 4 6 theme human 939:943 arg1 structure 966:974 the corresponding wild-type human acetylcholinesterase structure 911:974 the corresponding wild-type human acetylcholinesterase structure 911:974 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 5 7 theme neutral 1145:1151 arg1 moiety 1163:1168 a neutral isosteric moiety 1143:1168 a neutral isosteric moiety 1143:1168 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 4 8 theme complex 877:883 arg1 structure 847:855 The structure 843:855 The structure of the E202Q mutant complex 843:883 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 5 9 theme protein 1106:1112 arg1 core 1114:1117 the protein core 1102:1117 the protein core 1102:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 5 10 theme active 1222:1227 arg1 centre 1229:1234 the active centre 1218:1234 the active centre 1218:1234 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 0 11 theme toxin 108:112 arg1 fasciculin-II 114:126 the snake-venom toxin fasciculin-II 92:126 the snake-venom toxin fasciculin-II 92:126 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 5 12 theme isosteric 1153:1161 arg1 moiety 1163:1168 a neutral isosteric moiety 1143:1168 a neutral isosteric moiety 1143:1168 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 7 13 from role 1692:1695 arg1 interaction 1704:1714 the interaction 1700:1714 the interaction of acetylcholinesterase with substrates and inhibitors 1700:1769 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 5 14 theme centre 1229:1234 arg1 architecture 1202:1213 the functional architecture 1187:1213 the functional architecture of the active centre 1187:1234 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 0 15 theme snake-venom 96:106 arg1 fasciculin-II 114:126 the snake-venom toxin fasciculin-II 92:126 the snake-venom toxin fasciculin-II 92:126 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 6 16 dep residues 1329:1336 arg1 Tyr133 1354:1359 Tyr133 1354:1359 Tyr133 1354:1359 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 16 dep residues 1329:1336 arg1 residues 1329:1336 residues Glu202, Glu450, Tyr133 and two bridging molecules 1329:1386 residues Glu202, Glu450, Tyr133 and two bridging molecules 1329:1386 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 16 dep residues 1329:1336 arg1 Glu450 1346:1351 Glu450 1346:1351 Glu450 1346:1351 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 16 dep residues 1329:1336 arg1 molecules 1378:1386 two bridging molecules 1365:1386 two bridging molecules 1365:1386 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 16 dep residues 1329:1336 arg1 Glu202 1338:1343 Glu202 1338:1343 Glu202 1338:1343 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 7 17 theme main 1552:1555 arg1 role 1557:1560 the main role 1548:1560 the main role of this network 1548:1576 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 17 theme main 1552:1555 arg1 positioning 1592:1602 the proper positioning 1581:1602 the proper positioning of the Glu202 carboxylate relative to the catalytic triad 1581:1660 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 4 18 theme A 991:991 arg1 data 1004:1007 the 2.7 A resolution data 983:1007 the 2.7 A resolution data set collected 983:1021 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 2 19 theme mouse 644:648 arg1 acetylcholinesterase 650:669 mouse acetylcholinesterase 644:669 mouse acetylcholinesterase 644:669 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 7 20 theme proper 1585:1590 arg1 role 1557:1560 the main role 1548:1560 the main role of this network 1548:1576 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 20 theme proper 1585:1590 arg1 positioning 1592:1602 the proper positioning 1581:1602 the proper positioning of the Glu202 carboxylate relative to the catalytic triad 1581:1660 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 2 21 theme complex 397:403 arg1 structure 380:388 The structure 376:388 The structure of the complex of the wild-type enzyme 376:427 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 3 22 theme T. 733:734 arg1 enzyme 748:753 the T. californica enzyme 729:753 the T. californica enzyme with fasciculin-II 729:772 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 6 23 theme architecture 1265:1276 arg1 elements 1248:1255 the elements 1244:1255 the elements of this architecture 1244:1276 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 5 24 from substitution 1127:1138 arg1 core 1114:1117 the protein core 1102:1117 the protein core 1102:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 1 25 theme E202Q 203:207 arg1 mutant 209:214 its E202Q mutant 199:214 its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), 199:360 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 3 26 theme californica 736:746 arg1 enzyme 748:753 the T. californica enzyme 729:753 the T. californica enzyme with fasciculin-II 729:772 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 4 27 theme mutant 870:875 arg1 complex 877:883 the E202Q mutant complex 860:883 the E202Q mutant complex 860:883 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 7 28 with consistent 1516:1525 arg1 notion 1536:1541 the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors 1532:1769 the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors 1532:1769 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 1 29 theme mutant 209:214 arg1 Structures 129:138 Structures 129:138 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), 129:360 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 2 30 theme Torpedo 531:537 arg1 acetylcholinesterase 551:570 Torpedo californica acetylcholinesterase 531:570 Torpedo californica acetylcholinesterase 531:570 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 7 31 theme relative 1630:1637 arg1 role 1557:1560 the main role 1548:1560 the main role of this network 1548:1576 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 31 theme relative 1630:1637 arg1 positioning 1592:1602 the proper positioning 1581:1602 the proper positioning of the Glu202 carboxylate relative to the catalytic triad 1581:1660 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 2 32 theme similar 623:629 arg1 complex 631:637 a similar complex 621:637 a similar complex with mouse acetylcholinesterase 621:669 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 2 33 with complex 520:526 arg1 fasciculin-II 577:589 fasciculin-II 577:589 fasciculin-II 577:589 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 2 34 with complex 631:637 arg1 acetylcholinesterase 650:669 mouse acetylcholinesterase 644:669 mouse acetylcholinesterase 644:669 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 0 35 theme native 26:31 arg1 acetylcholinesterase 56:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 5 36 theme functional 1191:1200 arg1 architecture 1202:1213 the functional architecture 1187:1213 the functional architecture of the active centre 1187:1234 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 2 37 theme acetylcholinesterase 551:570 arg1 complex 520:526 the complex 516:526 the complex of Torpedo californica acetylcholinesterase with fasciculin-II 516:589 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 2 38 theme complex 520:526 arg1 structure 503:511 the structure 499:511 the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II 499:589 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 0 39 theme recombinant 14:24 arg1 acetylcholinesterase 56:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 6 40 theme vertebrate 1426:1435 arg1 acetylcholinesterases 1437:1457 other vertebrate acetylcholinesterases 1420:1457 other vertebrate acetylcholinesterases 1420:1457 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 2 41 theme californica 539:549 arg1 acetylcholinesterase 551:570 Torpedo californica acetylcholinesterase 531:570 Torpedo californica acetylcholinesterase 531:570 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 5 42 theme group 1091:1095 arg1 removal 1068:1074 removal 1068:1074 removal of the charged group from the protein core 1068:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 5 42 theme group 1091:1095 arg1 substitution 1127:1138 its substitution 1123:1138 its substitution by a neutral isosteric moiety 1123:1168 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 7 43 theme catalytic 1646:1654 arg1 triad 1656:1660 the catalytic triad 1642:1660 the catalytic triad 1642:1660 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 2 44 theme wild-type 412:420 arg1 enzyme 422:427 the wild-type enzyme 408:427 the wild-type enzyme 408:427 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 2 45 theme molecular 463:471 arg1 replacement 473:483 molecular replacement 463:483 molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II 463:589 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 7 46 theme carboxylate 1618:1628 arg1 role 1557:1560 the main role 1548:1560 the main role of this network 1548:1576 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 46 theme carboxylate 1618:1628 arg1 positioning 1592:1602 the proper positioning 1581:1602 the proper positioning of the Glu202 carboxylate relative to the catalytic triad 1581:1660 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 1 47 with complexes 219:227 arg1 toxin 278:282 a 'three-finger' polypeptide toxin 249:282 a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps) 249:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 1 47 with complexes 219:227 arg1 fasciculin-II 234:246 fasciculin-II 234:246 fasciculin-II 234:246 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 3 48 theme overall 676:682 arg1 structure 684:692 The overall structure 672:692 The overall structure 672:692 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 3 48 theme overall 676:682 arg1 similar 710:716 similar 710:716 similar 710:716 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 6 49 theme elements 1248:1255 arg1 network 1311:1317 a hydrogen-bond network 1295:1317 a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme 1295:1488 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 49 theme elements 1248:1255 arg1 One 1237:1239 One 1237:1239 One 1237:1239 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 49 theme elements 1248:1255 arg1 elements 1248:1255 the elements 1244:1255 the elements of this architecture 1244:1276 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 7 50 theme acetylcholinesterase 1719:1738 arg1 interaction 1704:1714 the interaction 1700:1714 the interaction of acetylcholinesterase with substrates and inhibitors 1700:1769 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 1 51 theme eastern 315:321 arg1 mamba 329:333 the eastern green mamba 311:333 the eastern green mamba (Dendroaspis angusticeps) 311:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 1 51 theme eastern 315:321 arg1 angusticeps 348:358 Dendroaspis angusticeps 336:358 Dendroaspis angusticeps 336:358 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 5 52 theme structures 1046:1055 arg1 Comparison 1024:1033 Comparison 1024:1033 Comparison of the two structures 1024:1055 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 0 53 theme human 50:54 arg1 acetylcholinesterase 56:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 1 54 theme green 323:327 arg1 mamba 329:333 the eastern green mamba 311:333 the eastern green mamba (Dendroaspis angusticeps) 311:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 1 54 theme green 323:327 arg1 angusticeps 348:358 Dendroaspis angusticeps 336:358 Dendroaspis angusticeps 336:358 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 6 55 theme bridging 1369:1376 arg1 molecules 1378:1386 two bridging molecules 1365:1386 two bridging molecules 1365:1386 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 4 56 theme resolution 993:1002 arg1 data 1004:1007 the 2.7 A resolution data 983:1007 the 2.7 A resolution data set collected 983:1021 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 4 57 theme E202Q 864:868 arg1 complex 877:883 the E202Q mutant complex 860:883 the E202Q mutant complex 860:883 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 0 58 theme mutant 43:48 arg1 acetylcholinesterase 56:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 recombinant native and E202Q mutant human acetylcholinesterase 14:75 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 1 59 theme mamba 329:333 arg1 venom 302:306 the venom 298:306 the venom of the eastern green mamba (Dendroaspis angusticeps) 298:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 4 60 theme acetylcholinesterase 945:964 arg1 structure 966:974 the corresponding wild-type human acetylcholinesterase structure 911:974 the corresponding wild-type human acetylcholinesterase structure 911:974 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 4 61 theme wild-type 929:937 arg1 structure 966:974 the corresponding wild-type human acetylcholinesterase structure 911:974 the corresponding wild-type human acetylcholinesterase structure 911:974 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 1 62 theme recombinant 143:153 arg1 acetylcholinesterase 171:190 recombinant wild-type human acetylcholinesterase 143:190 recombinant wild-type human acetylcholinesterase 143:190 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 7 63 theme functional 1681:1690 arg1 role 1692:1695 its functional role 1677:1695 its functional role in the interaction of acetylcholinesterase with substrates and inhibitors 1677:1769 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 3 64 theme mouse 807:811 arg1 complex 834:840 the mouse acetylcholinesterase complex 803:840 the mouse acetylcholinesterase complex 803:840 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 4 65 theme corresponding 915:927 arg1 structure 966:974 the corresponding wild-type human acetylcholinesterase structure 911:974 the corresponding wild-type human acetylcholinesterase structure 911:974 The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. 11053835 7 66 theme present 1495:1501 arg1 consistent 1516:1525 consistent 1516:1525 consistent 1516:1525 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 66 theme present 1495:1501 arg1 findings 1503:1510 The present findings 1491:1510 The present findings 1491:1510 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 1 67 theme wild-type 155:163 arg1 acetylcholinesterase 171:190 recombinant wild-type human acetylcholinesterase 143:190 recombinant wild-type human acetylcholinesterase 143:190 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 3 68 with enzyme 748:753 arg1 fasciculin-II 760:772 fasciculin-II 760:772 fasciculin-II 760:772 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 3 69 theme acetylcholinesterase 813:832 arg1 complex 834:840 the mouse acetylcholinesterase complex 803:840 the mouse acetylcholinesterase complex 803:840 The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. 11053835 5 70 theme charged 1083:1089 arg1 group 1091:1095 the charged group 1079:1095 the charged group from the protein core 1079:1117 Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. 11053835 6 71 theme water 1391:1395 arg1 network 1311:1317 a hydrogen-bond network 1295:1317 a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme 1295:1488 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 71 theme water 1391:1395 arg1 One 1237:1239 One 1237:1239 One 1237:1239 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 6 71 theme water 1391:1395 arg1 elements 1248:1255 the elements 1244:1255 the elements of this architecture 1244:1276 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 2 72 theme enzyme 422:427 arg1 complex 397:403 the complex 393:403 the complex of the wild-type enzyme 393:427 The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. 11053835 1 73 theme human 165:169 arg1 acetylcholinesterase 171:190 recombinant wild-type human acetylcholinesterase 143:190 recombinant wild-type human acetylcholinesterase 143:190 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 6 74 theme human 1477:1481 arg1 enzyme 1483:1488 the human enzyme 1473:1488 the human enzyme 1473:1488 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 1 75 theme three-finger 252:263 arg1 toxin 278:282 a 'three-finger' polypeptide toxin 249:282 a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps) 249:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 1 75 theme three-finger 252:263 arg1 fasciculin-II 234:246 fasciculin-II 234:246 fasciculin-II 234:246 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 0 76 theme acetylcholinesterase 56:75 arg1 Structures 0:9 Structures 0:9 Structures of recombinant native and E202Q mutant human acetylcholinesterase 0:75 Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. 11053835 1 77 theme Dendroaspis 336:346 arg1 mamba 329:333 the eastern green mamba 311:333 the eastern green mamba (Dendroaspis angusticeps) 311:359 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 1 77 theme Dendroaspis 336:346 arg1 angusticeps 348:358 Dendroaspis angusticeps 336:358 Dendroaspis angusticeps 336:358 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 6 78 theme other 1420:1424 arg1 acetylcholinesterases 1437:1457 other vertebrate acetylcholinesterases 1420:1457 other vertebrate acetylcholinesterases 1420:1457 One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. 11053835 1 79 theme acetylcholinesterase 171:190 arg1 Structures 129:138 Structures 129:138 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), 129:360 Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. 11053835 7 80 theme network 1570:1576 arg1 role 1557:1560 the main role 1548:1560 the main role of this network 1548:1576 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 11053835 7 80 theme network 1570:1576 arg1 positioning 1592:1602 the proper positioning 1581:1602 the proper positioning of the Glu202 carboxylate relative to the catalytic triad 1581:1660 The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. 20408548 4 0 theme human 685:689 arg1 butyrylcholinesterase 691:711 tabun-inhibited human butyrylcholinesterase 669:711 tabun-inhibited human butyrylcholinesterase 669:711 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 5 1 theme fasciculin 778:787 arg1 binding 767:773 binding 767:773 binding of fasciculin II 767:790 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 5 1 theme fasciculin 778:787 arg1 aging 757:761 aging 757:761 aging 757:761 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 4 2 theme tabun-inhibited 669:683 arg1 butyrylcholinesterase 691:711 tabun-inhibited human butyrylcholinesterase 669:711 tabun-inhibited human butyrylcholinesterase 669:711 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 6 3 theme nerve 1003:1007 arg1 agent 1009:1013 a nerve agent 1001:1013 a nerve agent 1001:1013 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 6 4 theme inhibition 1052:1061 arg1 mechanisms 1073:1082 the inhibition and aging mechanisms 1048:1082 mechanisms 1073:1082 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 3 5 theme aging 342:346 arg1 reaction 348:355 The aging reaction 338:355 The aging reaction 338:355 The aging reaction may proceed via either dealkylation or deamidation, depending on the stereochemistry of the phosphoramidyl adduct. 20408548 4 6 theme mouse 717:721 arg1 acetylcholinesterase 723:742 mouse acetylcholinesterase 717:742 mouse acetylcholinesterase 717:742 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 5 7 theme additional 844:853 arg1 interactions 867:878 additional stabilizing interactions 844:878 additional stabilizing interactions between the two subdomains that face each other across the active site gorge 844:955 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 3 8 theme phosphoramidyl 449:462 arg1 adduct 464:469 the phosphoramidyl adduct 445:469 the phosphoramidyl adduct 445:469 The aging reaction may proceed via either dealkylation or deamidation, depending on the stereochemistry of the phosphoramidyl adduct. 20408548 5 9 theme active 939:944 arg1 gorge 951:955 the active site gorge 935:955 the active site gorge 935:955 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 6 10 theme structural 1090:1099 arg1 template 1101:1108 a structural template 1088:1108 a structural template for the design of molecules capable of reactivating aged hAChE 1088:1171 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 6 11 theme molecules 1128:1136 arg1 design 1118:1123 the design 1114:1123 the design of molecules capable of reactivating aged hAChE 1114:1171 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 1 12 theme rapid 177:181 arg1 phosphylation 183:195 rapid phosphylation 177:195 rapid phosphylation of the catalytic serine 177:219 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 5 13 theme improved 803:810 arg1 thermostability 812:826 an improved thermostability 800:826 an improved thermostability 800:826 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 0 14 theme Structural 0:9 arg1 evidence 11:18 Structural evidence 0:18 Structural evidence that human acetylcholinesterase 0:50 Structural evidence that human acetylcholinesterase inhibited by tabun ages through O-dealkylation. 20408548 2 15 theme " 295:295 arg1 enzyme 297:302 an "aged" enzyme 287:302 an "aged" enzyme 287:302 A time-dependent reaction occurs on the tabun adduct, leading to an "aged" enzyme, resistant to oxime reactivators. 20408548 0 16 theme human 25:29 arg1 acetylcholinesterase 31:50 human acetylcholinesterase 25:50 human acetylcholinesterase 25:50 Structural evidence that human acetylcholinesterase inhibited by tabun ages through O-dealkylation. 20408548 6 17 theme hAChE 982:986 arg1 structure 969:977 This first structure 958:977 This first structure of hAChE inhibited by a nerve agent 958:1013 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 5 18 theme site 946:949 arg1 gorge 951:955 the active site gorge 935:955 the active site gorge 935:955 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 2 19 theme aged 291:294 arg1 enzyme 297:302 an "aged" enzyme 287:302 an "aged" enzyme 287:302 A time-dependent reaction occurs on the tabun adduct, leading to an "aged" enzyme, resistant to oxime reactivators. 20408548 6 20 theme aging 1067:1071 arg1 mechanisms 1073:1082 the inhibition and aging mechanisms 1048:1082 mechanisms 1073:1082 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 1 21 theme warfare 111:117 arg1 agent 119:123 a warfare agent 109:123 a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine 109:219 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 1 21 theme warfare 111:117 arg1 Tabun 100:104 Tabun 100:104 Tabun 100:104 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 1 22 theme catalytic 204:212 arg1 serine 214:219 the catalytic serine 200:219 the catalytic serine 200:219 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 5 23 theme stabilizing 855:865 arg1 interactions 867:878 additional stabilizing interactions 844:878 additional stabilizing interactions between the two subdomains that face each other across the active site gorge 844:955 Noteworthy, aging and binding of fasciculin II lead to an improved thermostability, resulting from additional stabilizing interactions between the two subdomains that face each other across the active site gorge. 20408548 1 24 theme serine 214:219 arg1 phosphylation 183:195 rapid phosphylation 177:195 rapid phosphylation of the catalytic serine 177:219 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 4 25 theme tabun-hAChE 510:520 arg1 structure 492:500 the X-ray structure 482:500 the X-ray structure of aged tabun-hAChE complexed with fasciculin II 482:549 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 0 26 theme tabun 65:69 arg1 ages 71:74 tabun ages 65:74 tabun ages 65:74 Structural evidence that human acetylcholinesterase inhibited by tabun ages through O-dealkylation. 20408548 4 27 theme aged 505:508 arg1 tabun-hAChE 510:520 aged tabun-hAChE 505:520 aged tabun-hAChE complexed with fasciculin II 505:549 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 2 28 theme time-dependent 224:237 arg1 reaction 239:246 A time-dependent reaction 222:246 A time-dependent reaction 222:246 A time-dependent reaction occurs on the tabun adduct, leading to an "aged" enzyme, resistant to oxime reactivators. 20408548 2 29 theme oxime 318:322 arg1 reactivators 324:335 oxime reactivators 318:335 oxime reactivators 318:335 A time-dependent reaction occurs on the tabun adduct, leading to an "aged" enzyme, resistant to oxime reactivators. 20408548 6 30 theme capable 1138:1144 arg1 molecules 1128:1136 molecules 1128:1136 molecules capable of reactivating aged hAChE 1128:1171 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 6 31 theme first 963:967 arg1 structure 969:977 This first structure 958:977 This first structure of hAChE inhibited by a nerve agent 958:1013 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 2 32 theme resistant 305:313 arg1 enzyme 297:302 an "aged" enzyme 287:302 an "aged" enzyme 287:302 A time-dependent reaction occurs on the tabun adduct, leading to an "aged" enzyme, resistant to oxime reactivators. 20408548 6 33 theme aged 1162:1165 arg1 hAChE 1167:1171 aged hAChE 1162:1171 aged hAChE 1162:1171 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 4 34 with agreement 611:619 arg1 mechanism 636:644 the aging mechanism 626:644 the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase 626:742 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 2 35 theme tabun 262:266 arg1 adduct 268:273 the tabun adduct 258:273 the tabun adduct 258:273 A time-dependent reaction occurs on the tabun adduct, leading to an "aged" enzyme, resistant to oxime reactivators. 20408548 1 36 theme human 139:143 arg1 hAChE 167:171 hAChE 167:171 hAChE 167:171 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 1 36 theme human 139:143 arg1 acetylcholinesterase 145:164 human acetylcholinesterase 139:164 human acetylcholinesterase (hAChE) 139:172 Tabun is a warfare agent that inhibits human acetylcholinesterase (hAChE) by rapid phosphylation of the catalytic serine. 20408548 3 37 theme adduct 464:469 arg1 stereochemistry 426:440 the stereochemistry 422:440 the stereochemistry of the phosphoramidyl adduct 422:469 The aging reaction may proceed via either dealkylation or deamidation, depending on the stereochemistry of the phosphoramidyl adduct. 20408548 4 38 theme aging 630:634 arg1 mechanism 636:644 the aging mechanism 626:644 the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase 626:742 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 20408548 6 39 theme structural 1024:1033 arg1 insight 1035:1041 structural insight 1024:1041 structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE 1024:1171 This first structure of hAChE inhibited by a nerve agent provides structural insight into the inhibition and aging mechanisms and a structural template for the design of molecules capable of reactivating aged hAChE. 20408548 4 40 theme X-ray 486:490 arg1 structure 492:500 the X-ray structure 482:500 the X-ray structure of aged tabun-hAChE complexed with fasciculin II 482:549 We solved the X-ray structure of aged tabun-hAChE complexed with fasciculin II, and we show that aging proceeds through O-dealkylation, in agreement with the aging mechanism that we determined for tabun-inhibited human butyrylcholinesterase and mouse acetylcholinesterase. 17157876 5 0 theme CPN/E 1014:1018 arg1 members 1020:1026 these other CPN/E members 1002:1026 these other CPN/E members 1002:1026 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 3 1 theme recombinant 615:625 arg1 CPN1 676:679 human CPN1 670:679 human CPN1 670:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 3 1 theme recombinant 615:625 arg1 domain 660:665 the recombinant C-terminally truncated catalytic domain 611:665 the recombinant C-terminally truncated catalytic domain of human CPN1 611:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 1 2 theme C-terminal 392:401 arg1 residues 409:416 C-terminal basic residues 392:416 C-terminal basic residues 392:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 6 3 theme CPN1 1352:1355 arg1 pocket 1342:1347 the S1' pocket 1334:1347 the S1' pocket of CPN1 1334:1355 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 5 4 theme catalytic 1105:1113 arg1 center 1115:1120 the catalytic center 1101:1120 the catalytic center 1101:1120 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 4 5 theme pear-like 791:799 arg1 shape 801:805 a pear-like shape 789:805 a pear-like shape 789:805 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 3 6 theme catalytic 650:658 arg1 CPN1 676:679 human CPN1 670:679 human CPN1 670:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 3 6 theme catalytic 650:658 arg1 domain 660:665 the recombinant C-terminally truncated catalytic domain 611:665 the recombinant C-terminally truncated catalytic domain of human CPN1 611:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 2 7 theme catalytic 552:560 arg1 domain 569:574 a 48 to 55 kDa catalytic (CPN1) domain 537:574 a 48 to 55 kDa catalytic (CPN1) domain 537:574 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 8 8 theme central 1761:1767 arg1 repeat 1782:1787 the central leucine-rich repeat tandem 1757:1794 the central leucine-rich repeat tandem of the cognate CPN2 subunit 1757:1822 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 9 theme CPN/E 129:133 arg1 subfamily 135:143 the CPN/E subfamily 125:143 the CPN/E subfamily of "regulatory" metallo-carboxypeptidases 125:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 3 10 theme CPN1 676:679 arg1 CPN1 676:679 human CPN1 670:679 human CPN1 670:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 3 10 theme CPN1 676:679 arg1 domain 660:665 the recombinant C-terminally truncated catalytic domain 611:665 the recombinant C-terminally truncated catalytic domain of human CPN1 611:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 2 11 theme 83 499:500 arg1 kDa 502:504 kDa 502:504 kDa 502:504 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 4 12 theme 319 824:826 arg1 residue 828:834 residue 828:834 residue 828:834 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 7 13 theme basic 1642:1646 arg1 residues 1648:1655 basic residues 1642:1655 basic residues probably involved in membrane interaction 1642:1697 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 6 14 theme Pro-Phe-Arg 1239:1249 arg1 end 1262:1264 the Pro-Phe-Arg C-terminal end 1235:1264 the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin 1235:1300 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 1 15 theme regulatory 149:158 arg1 metallo-carboxypeptidases 161:185 "regulatory" metallo-carboxypeptidases 148:185 "regulatory" metallo-carboxypeptidases 148:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 7 16 theme TT 1519:1520 arg1 edge 1522:1525 the distal TT edge 1508:1525 the distal TT edge of CPN1 1508:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 4 17 theme N-terminal 836:845 arg1 domain 857:862 a 319 residue N-terminal catalytic domain 822:862 a 319 residue N-terminal catalytic domain 822:862 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 8 18 theme cognate 1803:1809 arg1 subunit 1816:1822 the cognate CPN2 subunit 1799:1822 the cognate CPN2 subunit 1799:1822 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 19 theme factors 345:351 arg1 activity 297:304 the activity 293:304 the activity of vasoactive peptide hormones, growth factors and cytokines 293:365 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 20 theme metallo-carboxypeptidases 161:185 arg1 subfamily 135:143 the CPN/E subfamily 125:143 the CPN/E subfamily of "regulatory" metallo-carboxypeptidases 125:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 8 21 theme unique 1830:1835 arg1 patch 1857:1861 a unique hydrophobic surface patch 1828:1861 a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface 1828:1911 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 8 22 theme subunit 1816:1822 arg1 repeat 1782:1787 the central leucine-rich repeat tandem 1757:1794 the central leucine-rich repeat tandem of the cognate CPN2 subunit 1757:1822 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 23 theme cytokines 357:365 arg1 activity 297:304 the activity 293:304 the activity of vasoactive peptide hormones, growth factors and cytokines 293:365 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 7 24 used occupied 1630:1637 arg2 sites 1591:1595 equivalent sites 1580:1595 equivalent sites in the membrane-anchored CPM 1580:1624 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 4 25 dep abutting 871:878 arg1 shaped 895:900 shaped 895:900 shaped 895:900 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 8 26 theme patch 1857:1861 arg1 wrapping 1863:1870 a unique hydrophobic surface patch wrapping 1828:1870 a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface 1828:1911 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 6 27 theme C-terminal 1461:1470 arg1 residues 1476:1483 C-terminal Lys residues 1461:1483 C-terminal Lys residues 1461:1483 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 7 28 theme membrane-anchored 1604:1620 arg1 CPM 1622:1624 the membrane-anchored CPM 1600:1624 the membrane-anchored CPM 1600:1624 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 0 29 theme kininase 51:58 arg1 domain 73:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 2 30 theme blood 447:451 arg1 plasma 453:458 blood plasma 447:458 blood plasma 447:458 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 6 31 theme substrate 1281:1289 arg1 bradykinin 1291:1300 the natural substrate bradykinin 1269:1300 the natural substrate bradykinin 1269:1300 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 7 32 link O-linked 1539:1546 arg1 sugars 1572:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 7 32 link O-linked 1539:1546 arg1 residues 1496:1503 Three Thr residues 1486:1503 Three Thr residues at the distal TT edge of CPN1 1486:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 7 33 theme equivalent 1580:1589 arg1 sites 1591:1595 equivalent sites 1580:1595 equivalent sites in the membrane-anchored CPM 1580:1624 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 5 34 theme protein 1167:1173 arg1 inhibitors 1192:1201 some larger protein carboxypeptidase inhibitors 1155:1201 some larger protein carboxypeptidase inhibitors 1155:1201 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 3 35 theme crystal 724:730 arg1 structure 732:740 A crystal structure 722:740 its 2.1 A crystal structure 714:740 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 0 36 theme domain 73:78 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain 0:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 6 37 theme end 1262:1264 arg1 Modeling 1223:1230 Modeling 1223:1230 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site 1223:1321 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 1 38 theme peptide 320:326 arg1 hormones 328:335 vasoactive peptide hormones 309:335 vasoactive peptide hormones 309:335 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 8 39 theme active 1931:1936 arg1 centers 1938:1944 the two active centers 1923:1944 the two active centers 1923:1944 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 7 40 theme N-acetyl 1551:1558 arg1 glucosamine 1560:1570 N-acetyl glucosamine 1551:1570 N-acetyl glucosamine 1551:1570 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 0 41 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain 0:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 7 42 theme O-linked 1539:1546 arg1 sugars 1572:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 7 42 theme O-linked 1539:1546 arg1 residues 1496:1503 Three Thr residues 1486:1503 Three Thr residues at the distal TT edge of CPN1 1486:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 1 43 theme carboxypeptidase 87:102 arg1 CPN 107:109 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 43 theme carboxypeptidase 87:102 arg1 member 115:120 a member 113:120 a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases 113:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 43 theme carboxypeptidase 87:102 arg1 glycoprotein 208:219 an extracellular glycoprotein 191:219 an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues 191:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 7 44 theme CPN1 1530:1533 arg1 edge 1522:1525 the distal TT edge 1508:1525 the distal TT edge of CPN1 1508:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 4 45 theme residue 905:911 arg1 domain 957:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 0 46 theme human 25:29 arg1 domain 73:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 4 47 theme beta-sandwich 924:936 arg1 domain 957:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 8 48 theme hydrophobic 1837:1847 arg1 patch 1857:1861 a unique hydrophobic surface patch 1828:1861 a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface 1828:1911 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 0 49 theme N 48:48 arg1 domain 73:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 4 50 theme structural 747:756 arg1 analysis 758:765 The structural analysis 743:765 The structural analysis 743:765 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 6 51 theme bradykinin 1291:1300 arg1 end 1262:1264 the Pro-Phe-Arg C-terminal end 1235:1264 the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin 1235:1300 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 2 52 dep domains 513:519 arg1 each 521:524 each 521:524 each 521:524 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 7 53 theme membrane 1678:1685 arg1 interaction 1687:1697 membrane interaction 1678:1697 membrane interaction 1678:1697 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 1 54 theme basic 403:407 arg1 residues 409:416 C-terminal basic residues 392:416 C-terminal basic residues 392:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 7 55 from sites 1591:1595 arg1 CPM 1622:1624 the membrane-anchored CPM 1600:1624 the membrane-anchored CPM 1600:1624 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 8 56 theme leucine-rich 1769:1780 arg1 repeat 1782:1787 the central leucine-rich repeat tandem 1757:1794 the central leucine-rich repeat tandem of the cognate CPN2 subunit 1757:1822 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 2 57 dep kDa 502:504 arg1 CPN2 507:510 CPN2 507:510 CPN2 507:510 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 2 58 theme kDa 548:550 arg1 domain 569:574 a 48 to 55 kDa catalytic (CPN1) domain 537:574 a 48 to 55 kDa catalytic (CPN1) domain 537:574 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 3 59 theme truncated 640:648 arg1 CPN1 676:679 human CPN1 670:679 human CPN1 670:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 3 59 theme truncated 640:648 arg1 domain 660:665 the recombinant C-terminally truncated catalytic domain 611:665 the recombinant C-terminally truncated catalytic domain of human CPN1 611:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 8 60 theme tandem 1789:1794 arg1 repeat 1782:1787 the central leucine-rich repeat tandem 1757:1794 the central leucine-rich repeat tandem of the cognate CPN2 subunit 1757:1822 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 61 theme subfamily 135:143 arg1 CPN 107:109 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 61 theme subfamily 135:143 arg1 member 115:120 a member 113:120 a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases 113:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 6 62 theme C-terminal 1251:1260 arg1 end 1262:1264 the Pro-Phe-Arg C-terminal end 1235:1264 the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin 1235:1300 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 2 63 dep kDa 548:550 arg1 to 542:543 to 542:543 to 542:543 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 7 64 theme distal 1512:1517 arg1 edge 1522:1525 the distal TT edge 1508:1525 the distal TT edge of CPN1 1508:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 2 65 theme kDa 502:504 arg1 domains 513:519 two 83 kDa (CPN2) domains 495:519 two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain 495:574 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 3 66 theme human 670:674 arg1 CPN1 676:679 human CPN1 670:679 human CPN1 670:679 We have prepared and crystallized the recombinant C-terminally truncated catalytic domain of human CPN1, and have determined and refined its 2.1 A crystal structure. 17157876 8 67 theme CPN2 1811:1814 arg1 subunit 1816:1822 the cognate CPN2 subunit 1799:1822 the cognate CPN2 subunit 1799:1822 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 68 theme growth 338:343 arg1 factors 345:351 growth factors 338:351 growth factors 338:351 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 69 theme " 159:159 arg1 metallo-carboxypeptidases 161:185 "regulatory" metallo-carboxypeptidases 148:185 "regulatory" metallo-carboxypeptidases 148:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 4 70 theme residue 828:834 arg1 domain 857:862 a 319 residue N-terminal catalytic domain 822:862 a 319 residue N-terminal catalytic domain 822:862 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 1 71 theme N 104:104 arg1 CPN 107:109 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 71 theme N 104:104 arg1 member 115:120 a member 113:120 a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases 113:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 71 theme N 104:104 arg1 glycoprotein 208:219 an extracellular glycoprotein 191:219 an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues 191:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 7 72 theme Thr 1492:1494 arg1 sugars 1572:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 7 72 theme Thr 1492:1494 arg1 residues 1496:1503 Three Thr residues 1486:1503 Three Thr residues at the distal TT edge of CPN1 1486:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 5 73 theme carboxypeptidase 1175:1190 arg1 inhibitors 1192:1201 some larger protein carboxypeptidase inhibitors 1155:1201 some larger protein carboxypeptidase inhibitors 1155:1201 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 6 74 with agreement 1412:1420 arg1 preference 1433:1442 CPN's preference 1427:1442 CPN's preference for cleaving off C-terminal Lys residues 1427:1483 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 6 75 theme Lys 1472:1474 arg1 residues 1476:1483 C-terminal Lys residues 1461:1483 C-terminal Lys residues 1461:1483 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 1 76 dep glycoprotein 208:219 arg1 secreted 250:257 secreted 250:257 secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues 250:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 76 dep glycoprotein 208:219 arg1 synthesized 221:231 synthesized 221:231 synthesized in the liver 221:244 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 8 77 theme surface 1849:1855 arg1 patch 1857:1861 a unique hydrophobic surface patch 1828:1861 a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface 1828:1911 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 4 78 theme abutting 871:878 arg1 domain 957:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 0 79 theme catalytic 63:71 arg1 domain 73:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 5 80 theme larger 1160:1165 arg1 inhibitors 1192:1201 some larger protein carboxypeptidase inhibitors 1155:1201 some larger protein carboxypeptidase inhibitors 1155:1201 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 6 81 theme natural 1273:1279 arg1 bradykinin 1291:1300 the natural substrate bradykinin 1269:1300 the natural substrate bradykinin 1269:1300 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 5 82 theme surface 1033:1039 arg1 loops 1041:1045 two surface loops 1029:1045 two surface loops surrounding the active-site groove 1029:1080 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 4 83 contain has 785:787 arg2 shape 801:805 a pear-like shape 789:805 a pear-like shape 789:805 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 4 83 contain has 785:787 arg1 CPN1 780:783 CPN1 780:783 CPN1 780:783 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 4 84 theme catalytic 847:855 arg1 domain 857:862 a 319 residue N-terminal catalytic domain 822:862 a 319 residue N-terminal catalytic domain 822:862 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 2 85 theme CPN1 563:566 arg1 domain 569:574 a 48 to 55 kDa catalytic (CPN1) domain 537:574 a 48 to 55 kDa catalytic (CPN1) domain 537:574 Normally, CPN circulates in blood plasma as a hetero-tetramer consisting of two 83 kDa (CPN2) domains each flanked by a 48 to 55 kDa catalytic (CPN1) domain. 17157876 8 86 theme domain-TT 1893:1901 arg1 interface 1903:1911 the catalytic domain-TT interface 1879:1911 the catalytic domain-TT interface 1879:1911 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 87 theme vasoactive 309:318 arg1 hormones 328:335 vasoactive peptide hormones 309:335 vasoactive peptide hormones 309:335 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 8 88 theme tetrameric 1703:1712 arg1 CPN 1714:1716 tetrameric CPN 1703:1716 tetrameric CPN 1703:1716 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 89 theme hormones 328:335 arg1 activity 297:304 the activity 293:304 the activity of vasoactive peptide hormones, growth factors and cytokines 293:365 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 7 90 from edge 1522:1525 arg1 sugars 1572:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 O-linked to N-acetyl glucosamine sugars 1539:1577 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 7 90 from edge 1522:1525 arg1 residues 1496:1503 Three Thr residues 1486:1503 Three Thr residues at the distal TT edge of CPN1 1486:1533 Three Thr residues at the distal TT edge of CPN1 are O-linked to N-acetyl glucosamine sugars; equivalent sites in the membrane-anchored CPM are occupied by basic residues probably involved in membrane interaction. 17157876 1 91 gly glycoprotein 208:219 arg1 CPN 107:109 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 91 gly glycoprotein 208:219 arg1 glycoprotein 208:219 an extracellular glycoprotein 191:219 an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues 191:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 4 92 theme transthyretin 938:950 arg1 domain 957:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 6 93 theme S1 1338:1339 arg1 pocket 1342:1347 the S1' pocket 1334:1347 the S1' pocket of CPN1 1334:1355 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 8 94 theme CPN1 1724:1727 arg1 subunit 1729:1735 each CPN1 subunit 1719:1735 each CPN1 subunit 1719:1735 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 1 95 theme Human 81:85 arg1 CPN 107:109 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 95 theme Human 81:85 arg1 member 115:120 a member 113:120 a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases 113:185 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 95 theme Human 81:85 arg1 glycoprotein 208:219 an extracellular glycoprotein 191:219 an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues 191:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 4 96 theme TT 953:954 arg1 domain 957:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 0 97 theme carboxypeptidase 31:46 arg1 domain 73:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 the human carboxypeptidase N (kininase I) catalytic domain 21:78 Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. 17157876 4 98 theme 79 902:903 arg1 residue 905:911 residue 905:911 residue 905:911 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 8 99 theme catalytic 1883:1891 arg1 interface 1903:1911 the catalytic domain-TT interface 1879:1911 the catalytic domain-TT interface 1879:1911 In tetrameric CPN, each CPN1 subunit might interact with the central leucine-rich repeat tandem of the cognate CPN2 subunit via a unique hydrophobic surface patch wrapping around the catalytic domain-TT interface, exposing the two active centers. 17157876 5 100 theme other 1008:1012 arg1 members 1020:1026 these other CPN/E members 1002:1026 these other CPN/E members 1002:1026 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 4 101 theme C-terminal 913:922 arg1 domain 957:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain 868:962 The structural analysis reveals that CPN1 has a pear-like shape, consisting of a 319 residue N-terminal catalytic domain and an abutting, cylindrically shaped 79 residue C-terminal beta-sandwich transthyretin (TT) domain, more resembling CPD-2 than CPM. 17157876 6 102 theme Arg 1395:1397 arg1 residues 1399:1406 Arg residues 1395:1406 Arg residues 1395:1406 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 5 103 theme active-site 1063:1073 arg1 groove 1075:1080 the active-site groove 1059:1080 the active-site groove 1059:1080 Like these other CPN/E members, two surface loops surrounding the active-site groove restrict access to the catalytic center, offering an explanation for why some larger protein carboxypeptidase inhibitors do not inhibit CPN. 17157876 6 104 theme active 1311:1316 arg1 site 1318:1321 the active site 1307:1321 the active site 1307:1321 Modeling of the Pro-Phe-Arg C-terminal end of the natural substrate bradykinin into the active site shows that the S1' pocket of CPN1 might better accommodate P1'-Lys than Arg residues, in agreement with CPN's preference for cleaving off C-terminal Lys residues. 17157876 1 105 theme extracellular 194:206 arg1 CPN 107:109 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN) 81:110 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 17157876 1 105 theme extracellular 194:206 arg1 glycoprotein 208:219 an extracellular glycoprotein 191:219 an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues 191:416 Human carboxypeptidase N (CPN), a member of the CPN/E subfamily of "regulatory" metallo-carboxypeptidases, is an extracellular glycoprotein synthesized in the liver and secreted into the blood, where it controls the activity of vasoactive peptide hormones, growth factors and cytokines by specifically removing C-terminal basic residues. 9767079 3 0 theme methotrexate 619:630 arg1 uptake 632:637 methotrexate uptake 619:637 methotrexate uptake 619:637 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 2 1 from RFC 457:459 arg1 blots 472:476 western blots 464:476 western blots 464:476 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 3 2 dep decreased 601:609 arg1 % 599:599 approximately 25% 583:599 approximately 25% 583:599 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 8 3 theme transfected 1644:1654 arg1 cells 1656:1660 the transfected cells 1640:1660 the transfected cells 1640:1660 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 3 4 theme 3 492:492 arg1 microg/ml 494:502 microg/ml 494:502 microg/ml 494:502 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 4 5 theme 65 771:772 arg1 kDa 774:776 kDa 774:776 kDa 774:776 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 1 6 theme N-glycosylation 153:167 arg1 role 145:148 The role 141:148 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting 141:232 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 8 7 theme transport 1447:1455 arg1 activity 1457:1464 transport activity 1447:1464 transport activity 1447:1464 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 1 8 theme reduced 172:178 arg1 transport 201:209 reduced folate carrier (RFC) transport 172:209 reduced folate carrier (RFC) transport 172:209 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 6 9 theme carboxyl 1071:1078 arg1 terminus 1080:1087 the carboxyl terminus 1067:1087 the carboxyl terminus 1067:1087 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 6 10 dep functional 1104:1113 arg1 localized 1200:1208 localized 1200:1208 localized 1200:1208 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 1 11 theme carrier 187:193 arg1 transport 201:209 reduced folate carrier (RFC) transport 172:209 reduced folate carrier (RFC) transport 172:209 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 7 12 theme transport 1339:1347 arg1 function 1349:1356 transport function 1339:1356 transport function 1339:1356 Collectively, our results demonstrate that N-glycosylation of human RFC plays no significant role in either transport function or membrane targeting. 9767079 0 13 theme reduced 117:123 arg1 carrier 132:138 the human reduced folate carrier 107:138 the human reduced folate carrier 107:138 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 8 14 theme RFC 1428:1430 arg1 expression 1432:1441 RFC expression 1428:1441 RFC expression 1428:1441 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 6 15 theme anti-HA 1177:1183 arg1 antibody 1185:1192 rhodamine-conjugated anti-HA antibody 1156:1192 rhodamine-conjugated anti-HA antibody 1156:1192 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 4 16 theme RFC 657:659 arg1 protein 778:784 a 65 kDa protein 769:784 a 65 kDa protein 769:784 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 16 theme RFC 657:659 arg1 capacity 809:816 transport capacity 799:816 transport capacity 799:816 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 16 theme RFC 657:659 arg1 construct 666:674 A deglycosylated RFC cDNA construct 640:674 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) 640:735 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 0 17 theme carrier 132:138 arg1 activity 69:76 transport activity 59:76 transport activity 59:76 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 0 17 theme carrier 132:138 arg1 localization 91:102 cellular localization 82:102 cellular localization 82:102 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 3 18 theme complete 528:535 arg1 loss 537:540 the nearly complete loss 517:540 the nearly complete loss of glycosylated RFC 517:560 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 7 19 theme significant 1312:1322 arg1 role 1324:1327 no significant role 1309:1327 no significant role 1309:1327 Collectively, our results demonstrate that N-glycosylation of human RFC plays no significant role in either transport function or membrane targeting. 9767079 5 20 theme RFC 987:989 arg1 uptake 991:996 RFC uptake 987:996 RFC uptake 987:996 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 8 21 theme activity 1457:1464 arg1 stoichiometries 1409:1423 the stoichiometries 1405:1423 the stoichiometries of RFC expression and transport activity 1405:1464 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 6 22 theme immunofluorescence 1123:1140 arg1 staining 1142:1149 immunofluorescence staining 1123:1149 immunofluorescence staining with rhodamine-conjugated anti-HA antibody 1123:1192 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 1 23 theme human 306:310 arg1 cDNAs 316:320 human RFC cDNAs 306:320 human RFC cDNAs 306:320 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 3 24 theme RFC 558:560 arg1 loss 537:540 the nearly complete loss 517:540 the nearly complete loss of glycosylated RFC 517:560 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 0 25 theme transport 59:67 arg1 activity 69:76 transport activity 59:76 transport activity 59:76 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 0 26 from Effects 0:6 arg1 activity 69:76 transport activity 59:76 transport activity 59:76 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 0 26 from Effects 0:6 arg1 localization 91:102 cellular localization 82:102 cellular localization 82:102 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 5 27 theme Gln58-RFC 894:902 arg1 constructs 904:913 both wild-type and Gln58-RFC constructs 875:913 both wild-type and Gln58-RFC constructs 875:913 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 6 28 with staining 1142:1149 arg1 antibody 1185:1192 rhodamine-conjugated anti-HA antibody 1156:1192 rhodamine-conjugated anti-HA antibody 1156:1192 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 2 29 from shift 423:427 arg1 blots 472:476 western blots 464:476 western blots 464:476 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 7 30 theme RFC 1299:1301 arg1 N-glycosylation 1274:1288 N-glycosylation 1274:1288 N-glycosylation of human RFC 1274:1301 Collectively, our results demonstrate that N-glycosylation of human RFC plays no significant role in either transport function or membrane targeting. 9767079 2 31 theme progressive 411:421 arg1 shift 423:427 a progressive shift 409:427 a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa 409:486 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 1 32 theme transport-deficient 250:268 arg1 cells 283:287 transport-deficient K562 (K500E) cells 250:287 transport-deficient K562 (K500E) cells transfected with human RFC cDNAs 250:320 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 7 33 gly N-glycosylation 1274:1288 arg1 RFC 1299:1301 human RFC 1293:1301 human RFC 1293:1301 Collectively, our results demonstrate that N-glycosylation of human RFC plays no significant role in either transport function or membrane targeting. 9767079 8 34 theme regulatory 1526:1535 arg1 controls 1537:1544 identical regulatory controls 1516:1544 identical regulatory controls 1516:1544 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 5 35 theme wild-type 880:888 arg1 constructs 904:913 both wild-type and Gln58-RFC constructs 875:913 both wild-type and Gln58-RFC constructs 875:913 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 4 36 theme K500E 754:758 arg1 cells 760:764 K500E cells 754:764 K500E cells 754:764 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 6 37 from terminus 1080:1087 arg1 epitope 1056:1062 a hemagglutinin (HA) epitope 1035:1062 a hemagglutinin (HA) epitope at the carboxyl terminus 1035:1087 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 8 38 theme transport 1561:1569 arg1 components 1571:1580 non-RFC transport components 1553:1580 non-RFC transport components 1553:1580 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 5 39 theme relative 968:975 arg1 levels 977:982 relative levels 968:982 relative levels of RFC uptake 968:996 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 2 40 dep RFC 457:459 arg1 kDa 453:455 kDa 453:455 kDa 453:455 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 6 41 theme HA 1052:1053 arg1 epitope 1056:1062 a hemagglutinin (HA) epitope 1035:1062 a hemagglutinin (HA) epitope at the carboxyl terminus 1035:1087 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 3 42 theme decreased 601:609 arg1 rate 611:614 a approximately 25% decreased rate 581:614 a approximately 25% decreased rate of methotrexate uptake 581:637 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 5 43 theme RFC 943:945 arg1 protein 947:953 cDNA-encoded RFC protein 930:953 cDNA-encoded RFC protein 930:953 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 6 44 theme hemagglutinin 1037:1049 arg1 epitope 1056:1062 a hemagglutinin (HA) epitope 1035:1062 a hemagglutinin (HA) epitope at the carboxyl terminus 1035:1087 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 1 45 gly N-glycosylation 153:167 arg1 carrier 187:193 folate carrier 180:193 reduced folate carrier (RFC) transport 172:209 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 1 45 gly N-glycosylation 153:167 arg1 membrane 215:222 membrane targeting 215:232 membrane targeting 215:232 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 1 45 gly N-glycosylation 153:167 arg1 RFC 196:198 RFC 196:198 RFC 196:198 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 3 46 theme uptake 632:637 arg1 rate 611:614 a approximately 25% decreased rate 581:614 a approximately 25% decreased rate of methotrexate uptake 581:637 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 4 47 theme transport 799:807 arg1 construct 666:674 A deglycosylated RFC cDNA construct 640:674 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) 640:735 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 47 theme transport 799:807 arg1 protein 778:784 a 65 kDa protein 769:784 a 65 kDa protein 769:784 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 47 theme transport 799:807 arg1 capacity 809:816 transport capacity 799:816 transport capacity 799:816 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 6 48 theme plasma 1213:1218 arg1 membranes 1220:1228 plasma membranes 1213:1228 plasma membranes 1213:1228 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 4 49 theme kDa 774:776 arg1 protein 778:784 a 65 kDa protein 769:784 a 65 kDa protein 769:784 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 49 theme kDa 774:776 arg1 capacity 809:816 transport capacity 799:816 transport capacity 799:816 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 49 theme kDa 774:776 arg1 construct 666:674 A deglycosylated RFC cDNA construct 640:674 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) 640:735 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 3 50 gly glycosylated 545:556 arg1 RFC 558:560 glycosylated RFC 545:560 glycosylated RFC 545:560 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 0 51 theme human 111:115 arg1 carrier 132:138 the human reduced folate carrier 107:138 the human reduced folate carrier 107:138 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 3 52 theme microg/ml 494:502 arg1 tunicamycin 504:514 3 microg/ml tunicamycin 492:514 3 microg/ml tunicamycin 492:514 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 1 53 theme folate 180:185 arg1 carrier 187:193 folate carrier 180:193 reduced folate carrier (RFC) transport 172:209 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 1 53 theme folate 180:185 arg1 RFC 196:198 RFC 196:198 RFC 196:198 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 4 54 theme 6S 840:841 arg1 tetrahydrofolate 852:867 (6S)5-formyl tetrahydrofolate 839:867 (6S)5-formyl tetrahydrofolate 839:867 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 7 55 theme membrane 1361:1368 arg1 targeting 1370:1378 membrane targeting 1361:1378 membrane targeting 1361:1378 Collectively, our results demonstrate that N-glycosylation of human RFC plays no significant role in either transport function or membrane targeting. 9767079 0 56 theme folate 125:130 arg1 carrier 132:138 the human reduced folate carrier 107:138 the human reduced folate carrier 107:138 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 4 57 theme 5-formyl 843:850 arg1 tetrahydrofolate 852:867 (6S)5-formyl tetrahydrofolate 839:867 (6S)5-formyl tetrahydrofolate 839:867 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 8 58 theme expression 1432:1441 arg1 stoichiometries 1409:1423 the stoichiometries 1405:1423 the stoichiometries of RFC expression and transport activity 1405:1464 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 2 59 from blots 472:476 arg1 shift 423:427 a progressive shift 409:427 a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa 409:486 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 4 60 gly deglycosylated 642:655 arg1 protein 778:784 a 65 kDa protein 769:784 a 65 kDa protein 769:784 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 60 gly deglycosylated 642:655 arg1 capacity 809:816 transport capacity 799:816 transport capacity 799:816 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 60 gly deglycosylated 642:655 arg1 construct 666:674 A deglycosylated RFC cDNA construct 640:674 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) 640:735 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 6 61 contain containing 1024:1033 arg1 Wild-type 999:1007 Wild-type 999:1007 Wild-type 999:1007 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 6 61 contain containing 1024:1033 arg2 epitope 1056:1062 a hemagglutinin (HA) epitope 1035:1062 a hemagglutinin (HA) epitope at the carboxyl terminus 1035:1087 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 6 62 theme rhodamine-conjugated 1156:1175 arg1 antibody 1185:1192 rhodamine-conjugated anti-HA antibody 1156:1192 rhodamine-conjugated anti-HA antibody 1156:1192 Wild-type and Gln58-RFCs containing a hemagglutinin (HA) epitope at the carboxyl terminus were similarly functional and, by immunofluorescence staining with rhodamine-conjugated anti-HA antibody, were localized to plasma membranes. 9767079 4 63 theme deglycosylated 642:655 arg1 protein 778:784 a 65 kDa protein 769:784 a 65 kDa protein 769:784 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 63 theme deglycosylated 642:655 arg1 capacity 809:816 transport capacity 799:816 transport capacity 799:816 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 63 theme deglycosylated 642:655 arg1 construct 666:674 A deglycosylated RFC cDNA construct 640:674 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) 640:735 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 64 theme cDNA 661:664 arg1 protein 778:784 a 65 kDa protein 769:784 a 65 kDa protein 769:784 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 64 theme cDNA 661:664 arg1 capacity 809:816 transport capacity 799:816 transport capacity 799:816 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 4 64 theme cDNA 661:664 arg1 construct 666:674 A deglycosylated RFC cDNA construct 640:674 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) 640:735 A deglycosylated RFC cDNA construct in which asparagine-58 was replaced by glutamine (Gln58-RFC) was expressed in K500E cells as a 65 kDa protein and restored transport capacity for methotrexate and (6S)5-formyl tetrahydrofolate. 9767079 5 65 theme uptake 991:996 arg1 levels 977:982 relative levels 968:982 relative levels of RFC uptake 968:996 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 3 66 theme glycosylated 545:556 arg1 RFC 558:560 glycosylated RFC 545:560 glycosylated RFC 545:560 At 3 microg/ml tunicamycin, the nearly complete loss of glycosylated RFC was accompanied by a approximately 25% decreased rate of methotrexate uptake. 9767079 2 67 theme RFC 457:459 arg1 shift 423:427 a progressive shift 409:427 a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa 409:486 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 8 68 theme wild-type 1475:1483 arg1 RFC 1485:1487 wild-type RFC 1475:1487 wild-type RFC 1475:1487 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 1 69 theme RFC 312:314 arg1 cDNAs 316:320 human RFC cDNAs 306:320 human RFC cDNAs 306:320 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 0 70 theme cellular 82:89 arg1 localization 91:102 cellular localization 82:102 cellular localization 82:102 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 1 71 from role 145:148 arg1 transport 201:209 reduced folate carrier (RFC) transport 172:209 reduced folate carrier (RFC) transport 172:209 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 1 71 from role 145:148 arg1 targeting 224:232 membrane targeting 215:232 membrane targeting 215:232 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 7 72 theme human 1293:1297 arg1 RFC 1299:1301 human RFC 1293:1301 human RFC 1293:1301 Collectively, our results demonstrate that N-glycosylation of human RFC plays no significant role in either transport function or membrane targeting. 9767079 2 73 theme western 464:470 arg1 blots 472:476 western blots 464:476 western blots 464:476 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 8 74 theme identical 1516:1524 arg1 controls 1537:1544 identical regulatory controls 1516:1544 identical regulatory controls 1516:1544 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 0 75 dep activity 69:76 arg1 the 55:57 the 55:57 the 55:57 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 2 76 theme wild-type 353:361 arg1 RFC 363:365 wild-type RFC 353:365 wild-type RFC 353:365 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 0 77 theme loss 15:18 arg1 Effects 0:6 Effects 0:6 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 0:139 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 1 78 theme K562 270:273 arg1 cells 283:287 transport-deficient K562 (K500E) cells 250:287 transport-deficient K562 (K500E) cells transfected with human RFC cDNAs 250:320 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 8 79 theme non-RFC 1553:1559 arg1 components 1571:1580 non-RFC transport components 1553:1580 non-RFC transport components 1553:1580 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 0 80 theme capacity 23:30 arg1 loss 15:18 the loss 11:18 the loss of capacity for N-glycosylation 11:50 Effects of the loss of capacity for N-glycosylation on the transport activity and cellular localization of the human reduced folate carrier. 9767079 1 81 theme K500E 276:280 arg1 cells 283:287 transport-deficient K562 (K500E) cells 250:287 transport-deficient K562 (K500E) cells transfected with human RFC cDNAs 250:320 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 9767079 5 82 theme protein 947:953 arg1 expression 916:925 expression 916:925 expression of cDNA-encoded RFC protein 916:953 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 2 83 dep kDa 453:455 arg1 85 450:451 85 450:451 85 450:451 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 2 84 with Treatment 323:331 arg1 tunicamycin 372:382 tunicamycin 372:382 tunicamycin (0-3 microg) 372:395 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 2 84 with Treatment 323:331 arg1 microg 389:394 0-3 microg 385:394 0-3 microg 385:394 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 5 85 theme cDNA-encoded 930:941 arg1 protein 947:953 cDNA-encoded RFC protein 930:953 cDNA-encoded RFC protein 930:953 With both wild-type and Gln58-RFC constructs, expression of cDNA-encoded RFC protein far exceeded relative levels of RFC uptake. 9767079 8 86 theme transport 1618:1626 arg1 function 1628:1635 transport function 1618:1635 transport function 1618:1635 The discrepancy between the stoichiometries of RFC expression and transport activity for both wild-type RFC and Gln58-RFC implies that identical regulatory controls and/or non-RFC transport components are necessary to completely restore transport function in the transfected cells. 9767079 2 87 theme cells 336:340 arg1 Treatment 323:331 Treatment 323:331 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) 323:395 Treatment of cells expressing wild-type RFC with tunicamycin (0-3 microg) resulted in a progressive shift of the approximately 85 kDa RFC on western blots to 65 kDa. 9767079 1 88 theme membrane 215:222 arg1 targeting 224:232 membrane targeting 215:232 membrane targeting 215:232 The role of N-glycosylation in reduced folate carrier (RFC) transport and membrane targeting was examined in transport-deficient K562 (K500E) cells transfected with human RFC cDNAs. 8172892 0 0 contain has 38:40 arg2 oligosaccharides 42:57 oligosaccharides 42:57 oligosaccharides O-glycosidically linked to threonine residues at 159 and 169 42:118 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 0 0 contain has 38:40 arg1 peptide 11:17 Activation peptide 0:17 Activation peptide of human factor IX 0:36 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 3 1 theme AP 465:466 arg1 beta 468:471 AP beta 465:471 AP beta 465:471 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 9 2 from 169 1662:1664 arg1 residues 1642:1649 the threonine residues 1628:1649 the threonine residues at 159 and 169 1628:1664 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 6 3 theme D-galactose 1096:1106 arg1 mol 1049:1051 1 mol 1047:1051 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid 1047:1129 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 7 4 theme structures 1298:1307 arg1 existence 1218:1226 the existence 1214:1226 the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures 1214:1307 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 6 5 theme acid 1126:1129 arg1 mol 1049:1051 1 mol 1047:1051 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid 1047:1129 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 2 6 theme factor 270:275 arg1 IX 277:278 human factor IX 264:278 human factor IX 264:278 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 5 7 from present 887:893 arg1 alpha 901:905 AP alpha 898:905 AP alpha 898:905 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 9 8 from 159 1654:1656 arg1 residues 1642:1649 the threonine residues 1628:1649 the threonine residues at 159 and 169 1628:1664 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 4 9 theme AP 733:734 arg1 alpha 736:740 AP alpha 733:740 AP alpha 733:740 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 9 10 contain has 1571:1573 arg2 tetrasaccharides 1584:1599 tetrasaccharides 1584:1599 tetrasaccharides 1584:1599 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 9 10 contain has 1571:1573 arg2 tri- 1575:1578 tri- 1575:1578 tri- 1575:1578 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 9 10 contain has 1571:1573 arg1 part 1500:1503 a part 1498:1503 a part of the activation peptide of human factor IX in circulating blood 1498:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 1 11 theme activation 176:185 arg1 peptide 187:193 the activation peptide 172:193 the activation peptide (AP) of human blood coagulation factor IX 172:235 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 1 11 theme activation 176:185 arg1 AP 196:197 AP 196:197 AP 196:197 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 7 12 theme GalNAc-Thr 1260:1269 arg1 existence 1218:1226 the existence 1214:1226 the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures 1214:1307 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 9 13 theme peptide 1523:1529 arg1 part 1500:1503 a part 1498:1503 a part of the activation peptide of human factor IX in circulating blood 1498:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 7 14 theme Gal- 1255:1258 arg1 GalNAc-Thr 1260:1269 NeuNAc-(Gal-)GalNAc-Thr 1247:1269 NeuNAc-(Gal-)GalNAc-Thr 1247:1269 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 6 15 dep mol 1049:1051 arg1 each 1053:1056 each 1053:1056 each 1053:1056 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 9 16 theme human 1534:1538 arg1 IX 1547:1548 human factor IX 1534:1548 human factor IX in circulating blood 1534:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 7 17 theme NeuNAc- 1247:1253 arg1 GalNAc-Thr 1260:1269 NeuNAc-(Gal-)GalNAc-Thr 1247:1269 NeuNAc-(Gal-)GalNAc-Thr 1247:1269 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 1 18 theme human 203:207 arg1 IX 234:235 human blood coagulation factor IX 203:235 human blood coagulation factor IX 203:235 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 9 19 theme IX 1547:1548 arg1 peptide 1523:1529 the activation peptide 1508:1529 the activation peptide of human factor IX in circulating blood 1508:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 7 20 theme Gal-GalNAc-Thr 1231:1244 arg1 existence 1218:1226 the existence 1214:1226 the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures 1214:1307 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 0 21 from 159 108:110 arg1 residues 96:103 threonine residues 86:103 threonine residues at 159 and 169 86:118 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 9 22 theme circulating 1553:1563 arg1 blood 1565:1569 circulating blood 1553:1569 circulating blood 1553:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 8 23 theme amino 1326:1330 arg1 analysis 1337:1344 amino acid analysis 1326:1344 amino acid analysis after the isolation of AP alpha 1326:1376 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 1 24 theme oligosaccharide 130:144 arg1 chains 146:151 O-Linked oligosaccharide chains 121:151 O-Linked oligosaccharide chains 121:151 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 3 25 attach presence 562:569 arg1 molecule 608:615 the molecule 604:615 the molecule of AP alpha 604:627 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 25 attach presence 562:569 arg2 s 598:598 s 598:598 s 598:598 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 25 attach presence 562:569 arg2 chain 592:596 an O-linked sugar chain 574:596 an O-linked sugar chain(s) 574:599 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 9 26 from blood 1565:1569 arg1 peptide 1523:1529 the activation peptide 1508:1529 the activation peptide of human factor IX in circulating blood 1508:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 4 27 theme protein 828:834 arg1 sequencer 836:844 a gas-phase protein sequencer 816:844 a gas-phase protein sequencer 816:844 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 5 28 theme other 851:855 arg1 serine 857:862 All other serine and threonine residues 847:885 serine 857:862 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 5 29 attach present 887:893 arg1 alpha 901:905 AP alpha 898:905 AP alpha 898:905 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 5 29 attach present 887:893 arg2 serine 857:862 All other serine and threonine residues 847:885 serine 857:862 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 8 30 theme alpha 1372:1376 arg1 isolation 1356:1364 the isolation 1352:1364 the isolation of AP alpha 1352:1376 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 3 31 from presence 562:569 arg1 molecule 608:615 the molecule 604:615 the molecule of AP alpha 604:627 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 7 32 theme alpha-D4 1195:1202 arg1 analysis 1180:1187 Fast atom bombardment tandem mass spectrometric analysis 1132:1187 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 9 33 theme threonine 1632:1640 arg1 residues 1642:1649 the threonine residues 1628:1649 the threonine residues at 159 and 169 1628:1664 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 2 34 dep species 315:321 arg1 gamma 350:354 AP gamma 347:354 AP gamma 347:354 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 34 dep species 315:321 arg1 species 315:321 three molecular species 299:321 three molecular species (AP alpha, AP beta, and AP gamma) 299:355 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 34 dep species 315:321 arg1 alpha 327:331 AP alpha 324:331 AP alpha 324:331 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 34 dep species 315:321 arg1 beta 337:340 AP beta 334:340 AP beta 334:340 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 0 35 theme Activation 0:9 arg1 peptide 11:17 Activation peptide 0:17 Activation peptide of human factor IX 0:36 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 4 36 dep nonapeptide 632:642 arg1 residues 658:665 residues 157-165 658:673 residues 157-165 658:673 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 36 dep nonapeptide 632:642 arg1 alpha-D4 648:655 AP alpha-D4 645:655 AP alpha-D4 645:655 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 6 37 theme Component 946:954 arg1 sugar 956:960 Component sugar 946:960 Component sugar 946:960 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 2 38 theme high-performance 375:390 arg1 chromatography 399:412 reversed-phase high-performance liquid chromatography 360:412 reversed-phase high-performance liquid chromatography 360:412 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 3 39 theme AP 477:478 arg1 gamma 480:484 AP gamma 477:484 AP gamma 477:484 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 7 40 theme spectrometric 1166:1178 arg1 analysis 1180:1187 Fast atom bombardment tandem mass spectrometric analysis 1132:1187 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 3 41 theme acid 421:424 arg1 analysis 426:433 Amino acid analysis 415:433 Amino acid analysis 415:433 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 0 42 theme factor 28:33 arg1 IX 35:36 human factor IX 22:36 human factor IX 22:36 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 7 43 theme tandem 1154:1159 arg1 analysis 1180:1187 Fast atom bombardment tandem mass spectrometric analysis 1132:1187 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 3 44 theme sugar 586:590 arg1 chain 592:596 an O-linked sugar chain 574:596 an O-linked sugar chain(s) 574:599 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 44 theme sugar 586:590 arg1 s 598:598 s 598:598 s 598:598 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 2 45 theme AP 324:325 arg1 species 315:321 three molecular species 299:321 three molecular species (AP alpha, AP beta, and AP gamma) 299:355 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 45 theme AP 324:325 arg1 alpha 327:331 AP alpha 324:331 AP alpha 324:331 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 7 46 theme atom 1137:1140 arg1 bombardment 1142:1152 Fast atom bombardment 1132:1152 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 8 47 theme total 1421:1425 arg1 peptide 1438:1444 the total activation peptide 1417:1444 the total activation peptide obtained 1417:1453 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 3 48 theme AP 447:448 arg1 alpha 450:454 AP alpha 447:454 AP alpha 447:454 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 1 49 theme blood 209:213 arg1 IX 234:235 human blood coagulation factor IX 203:235 human blood coagulation factor IX 203:235 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 6 50 theme alpha-D4 993:1000 arg1 analyses 978:985 Component sugar and sialic acid analyses 946:985 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 946:1016 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 8 51 theme peptide 1438:1444 arg1 peptide 1438:1444 the total activation peptide 1417:1444 the total activation peptide obtained 1417:1453 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 8 51 theme peptide 1438:1444 arg1 % 1412:1412 approximately 35% 1396:1412 approximately 35% of the total activation peptide obtained 1396:1453 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 1 52 theme factor 227:232 arg1 IX 234:235 human blood coagulation factor IX 203:235 human blood coagulation factor IX 203:235 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 3 53 link O-linked 577:584 arg1 chain 592:596 an O-linked sugar chain 574:596 an O-linked sugar chain(s) 574:599 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 53 link O-linked 577:584 arg1 s 598:598 s 598:598 s 598:598 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 0 54 from 169 116:118 arg1 residues 96:103 threonine residues 86:103 threonine residues at 159 and 169 86:118 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 3 55 theme alpha 623:627 arg1 molecule 608:615 the molecule 604:615 the molecule of AP alpha 604:627 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 6 56 theme acid 973:976 arg1 analyses 978:985 Component sugar and sialic acid analyses 946:985 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 946:1016 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 6 57 contain contained 1037:1045 arg1 they 1032:1035 they 1032:1035 they 1032:1035 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 6 57 contain contained 1037:1045 arg2 mol 1049:1051 1 mol 1047:1051 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid 1047:1129 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 7 58 theme Fast 1132:1135 arg1 bombardment 1142:1152 Fast atom bombardment 1132:1152 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 6 59 theme sialic 1119:1124 arg1 acid 1126:1129 sialic acid 1119:1129 sialic acid 1119:1129 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 2 60 theme human 264:268 arg1 IX 277:278 human factor IX 264:278 human factor IX 264:278 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 7 61 theme NeuNAc-Gal-GalNAc-Thr 1276:1296 arg1 structures 1298:1307 NeuNAc-Gal-GalNAc-Thr structures 1276:1307 NeuNAc-Gal-GalNAc-Thr structures 1276:1307 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 3 62 contain contained 487:495 arg1 beta 468:471 AP beta 465:471 AP beta 465:471 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 62 contain contained 487:495 arg1 alpha 450:454 AP alpha 447:454 AP alpha 447:454 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 62 contain contained 487:495 arg2 galactosamine 497:509 galactosamine 497:509 galactosamine in addition to glucosamine 497:536 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 9 63 attach linked 1618:1623 arg1 residues 1642:1649 the threonine residues 1628:1649 the threonine residues at 159 and 169 1628:1664 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 9 63 attach linked 1618:1623 arg2 tri- 1575:1578 tri- 1575:1578 tri- 1575:1578 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 9 64 theme activation 1512:1521 arg1 peptide 1523:1529 the activation peptide 1508:1529 the activation peptide of human factor IX in circulating blood 1508:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 6 65 theme AP 1006:1007 arg1 alpha-D5 1009:1016 AP alpha-D5 1006:1016 AP alpha-D5 1006:1016 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 5 66 from alpha 901:905 arg1 present 887:893 present 887:893 present 887:893 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 9 67 theme factor 1540:1545 arg1 IX 1547:1548 human factor IX 1534:1548 human factor IX in circulating blood 1534:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 8 68 theme acid 1332:1335 arg1 analysis 1337:1344 amino acid analysis 1326:1344 amino acid analysis after the isolation of AP alpha 1326:1376 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 5 69 theme peptide 926:932 arg1 sequencing 934:943 peptide sequencing 926:943 peptide sequencing 926:943 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 1 70 theme O-Linked 121:128 arg1 chains 146:151 O-Linked oligosaccharide chains 121:151 O-Linked oligosaccharide chains 121:151 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 4 71 contain contained 742:750 arg1 AP 698:699 AP alpha-D5, 166-176 698:717 AP 698:699 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 71 contain contained 742:750 arg1 undecapeptide 683:695 an undecapeptide 680:695 an undecapeptide (AP alpha-D5, 166-176) 680:718 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 71 contain contained 742:750 arg2 Thr-169 764:770 Thr-169 764:770 Thr-169 764:770 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 71 contain contained 742:750 arg1 nonapeptide 632:642 A nonapeptide 630:642 A nonapeptide (AP alpha-D4, residues 157-165) 630:674 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 71 contain contained 742:750 arg2 Thr-159 752:758 Thr-159 752:758 Thr-159 752:758 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 9 72 from IX 1547:1548 arg1 blood 1565:1569 circulating blood 1553:1569 circulating blood 1553:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 6 73 theme N-acetyl-D-galactosamine 1061:1084 arg1 mol 1049:1051 1 mol 1047:1051 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid 1047:1129 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 4 74 attach derived 720:726 arg1 alpha 736:740 AP alpha 733:740 AP alpha 733:740 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 74 attach derived 720:726 arg2 nonapeptide 632:642 A nonapeptide 630:642 A nonapeptide (AP alpha-D4, residues 157-165) 630:674 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 0 75 theme threonine 86:94 arg1 residues 96:103 threonine residues 86:103 threonine residues at 159 and 169 86:118 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 8 76 theme AP 1369:1370 arg1 alpha 1372:1376 AP alpha 1369:1376 AP alpha 1369:1376 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 9 77 from peptide 1523:1529 arg1 blood 1565:1569 circulating blood 1553:1569 circulating blood 1553:1569 From these results, it was concluded that a part of the activation peptide of human factor IX in circulating blood has tri- and tetrasaccharides O-glycosidically linked to the threonine residues at 159 and 169. 8172892 4 78 theme gas-phase 818:826 arg1 sequencer 836:844 a gas-phase protein sequencer 816:844 a gas-phase protein sequencer 816:844 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 7 79 theme AP 1192:1193 arg1 alpha-D4 1195:1202 AP alpha-D4 1192:1202 AP alpha-D4 1192:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 3 80 theme Amino 415:419 arg1 analysis 426:433 Amino acid analysis 415:433 Amino acid analysis 415:433 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 6 81 theme sugar 956:960 arg1 analyses 978:985 Component sugar and sialic acid analyses 946:985 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 946:1016 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 4 82 theme AP 645:646 arg1 residues 658:665 residues 157-165 658:673 residues 157-165 658:673 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 4 82 theme AP 645:646 arg1 alpha-D4 648:655 AP alpha-D4 645:655 AP alpha-D4 645:655 A nonapeptide (AP alpha-D4, residues 157-165) and an undecapeptide (AP alpha-D5, 166-176) derived from AP alpha contained Thr-159 and Thr-169, neither of which could be identified using a gas-phase protein sequencer. 8172892 0 83 theme human 22:26 arg1 IX 35:36 human factor IX 22:36 human factor IX 22:36 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 8 84 dep analysis 1337:1344 arg1 basis 1317:1321 basis 1317:1321 basis 1317:1321 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 8 84 dep analysis 1337:1344 arg1 the 1313:1315 the 1313:1315 the 1313:1315 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 7 85 theme mass 1161:1164 arg1 analysis 1180:1187 Fast atom bombardment tandem mass spectrometric analysis 1132:1187 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 2 86 theme liquid 392:397 arg1 chromatography 399:412 reversed-phase high-performance liquid chromatography 360:412 reversed-phase high-performance liquid chromatography 360:412 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 3 87 theme chain 592:596 arg1 presence 562:569 the presence 558:569 the presence of an O-linked sugar chain(s) in the molecule of AP alpha 558:627 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 2 88 theme reversed-phase 360:373 arg1 chromatography 399:412 reversed-phase high-performance liquid chromatography 360:412 reversed-phase high-performance liquid chromatography 360:412 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 0 89 theme IX 35:36 arg1 peptide 11:17 Activation peptide 0:17 Activation peptide of human factor IX 0:36 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 5 90 theme AP 898:899 arg1 alpha 901:905 AP alpha 898:905 AP alpha 898:905 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 7 91 theme bombardment 1142:1152 arg1 analysis 1180:1187 Fast atom bombardment tandem mass spectrometric analysis 1132:1187 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 1132:1202 Fast atom bombardment tandem mass spectrometric analysis of AP alpha-D4 suggested the existence of Gal-GalNAc-Thr, NeuNAc-(Gal-)GalNAc-Thr, and NeuNAc-Gal-GalNAc-Thr structures. 8172892 6 92 theme sialic 966:971 arg1 acid 973:976 sialic acid 966:976 sialic acid 966:976 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 3 93 theme O-linked 577:584 arg1 chain 592:596 an O-linked sugar chain 574:596 an O-linked sugar chain(s) 574:599 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 3 93 theme O-linked 577:584 arg1 s 598:598 s 598:598 s 598:598 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 0 94 attach linked 76:81 arg2 oligosaccharides 42:57 oligosaccharides 42:57 oligosaccharides O-glycosidically linked to threonine residues at 159 and 169 42:118 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 0 94 attach linked 76:81 arg1 residues 96:103 threonine residues 86:103 threonine residues at 159 and 169 86:118 Activation peptide of human factor IX has oligosaccharides O-glycosidically linked to threonine residues at 159 and 169. 8172892 6 95 theme alpha-D5 1009:1016 arg1 analyses 978:985 Component sugar and sialic acid analyses 946:985 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 946:1016 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 8 96 theme activation 1427:1436 arg1 peptide 1438:1444 the total activation peptide 1417:1444 the total activation peptide obtained 1417:1453 On the basis of amino acid analysis after the isolation of AP alpha, it accounted for approximately 35% of the total activation peptide obtained. 8172892 5 97 theme threonine 868:876 arg1 residues 878:885 All other serine and threonine residues 847:885 residues 878:885 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 2 98 theme molecular 305:313 arg1 gamma 350:354 AP gamma 347:354 AP gamma 347:354 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 98 theme molecular 305:313 arg1 species 315:321 three molecular species 299:321 three molecular species (AP alpha, AP beta, and AP gamma) 299:355 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 98 theme molecular 305:313 arg1 alpha 327:331 AP alpha 324:331 AP alpha 324:331 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 98 theme molecular 305:313 arg1 beta 337:340 AP beta 334:340 AP beta 334:340 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 1 99 theme coagulation 215:225 arg1 IX 234:235 human blood coagulation factor IX 203:235 human blood coagulation factor IX 203:235 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 5 100 theme present 887:893 arg1 serine 857:862 All other serine and threonine residues 847:885 serine 857:862 All other serine and threonine residues present in AP alpha were identified by peptide sequencing. 8172892 6 101 theme AP 990:991 arg1 alpha-D4 993:1000 AP alpha-D4 990:1000 AP alpha-D4 990:1000 Component sugar and sialic acid analyses of AP alpha-D4 and AP alpha-D5 revealed that they contained 1 mol each of N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), and sialic acid. 8172892 2 102 theme AP 347:348 arg1 gamma 350:354 AP gamma 347:354 AP gamma 347:354 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 102 theme AP 347:348 arg1 species 315:321 three molecular species 299:321 three molecular species (AP alpha, AP beta, and AP gamma) 299:355 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 3 103 theme AP 620:621 arg1 alpha 623:627 AP alpha 620:627 AP alpha 620:627 Amino acid analysis showed that AP alpha, but not AP beta and AP gamma, contained galactosamine in addition to glucosamine, thereby suggesting the presence of an O-linked sugar chain(s) in the molecule of AP alpha. 8172892 2 104 theme AP 334:335 arg1 species 315:321 three molecular species 299:321 three molecular species (AP alpha, AP beta, and AP gamma) 299:355 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 2 104 theme AP 334:335 arg1 beta 337:340 AP beta 334:340 AP beta 334:340 The peptide obtained from human factor IX was separated into three molecular species (AP alpha, AP beta, and AP gamma) by reversed-phase high-performance liquid chromatography. 8172892 1 105 theme IX 234:235 arg1 peptide 187:193 the activation peptide 172:193 the activation peptide (AP) of human blood coagulation factor IX 172:235 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 8172892 1 105 theme IX 234:235 arg1 AP 196:197 AP 196:197 AP 196:197 O-Linked oligosaccharide chains were identified in the activation peptide (AP) of human blood coagulation factor IX. 21269602 0 0 theme classical 78:86 arg1 cadherin 88:95 a divergent classical cadherin 66:95 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 0:96 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 1 1 theme divergent 145:153 arg1 cadherin 119:126 Vascular endothelial cadherin 98:126 Vascular endothelial cadherin (VE-cadherin) 98:140 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 1 theme divergent 145:153 arg1 member 155:160 a divergent member 143:160 a divergent member of the type II classical cadherin family of cell adhesion proteins 143:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 2 2 with investigations 297:310 arg1 protein 340:346 a bacterially produced protein 317:346 a bacterially produced protein 317:346 Previous investigations with a bacterially produced protein suggested that VE-cadherin forms cell surface trimers that bind between apposed cells to form hexamers. 21269602 0 3 theme divergent 68:76 arg1 cadherin 88:95 a divergent classical cadherin 66:95 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 0:96 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 7 4 theme type 1291:1294 arg1 characteristics 1256:1270 characteristics 1256:1270 characteristics of both type I and type II subfamilies 1256:1309 Thus, VE-cadherin is an outlier among classical cadherins, with characteristics of both type I and type II subfamilies. 21269602 1 5 theme homophilic 239:248 arg1 adhesion 250:257 homophilic adhesion 239:257 homophilic adhesion 239:257 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 6 theme endothelial 107:117 arg1 VE-cadherin 129:139 VE-cadherin 129:139 VE-cadherin 129:139 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 6 theme endothelial 107:117 arg1 cadherin 119:126 Vascular endothelial cadherin 98:126 Vascular endothelial cadherin (VE-cadherin) 98:140 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 6 theme endothelial 107:117 arg1 member 155:160 a divergent member 143:160 a divergent member of the type II classical cadherin family of cell adhesion proteins 143:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 6 7 theme anchor 1047:1052 arg1 residues 1054:1061 two tryptophan anchor residues 1032:1061 two tryptophan anchor residues 1032:1061 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 4 8 theme glycosylation 765:777 arg1 lack 757:760 a lack 755:760 a lack of glycosylation 755:777 Trimerization of the bacterially produced protein appears to be an artifact that arises from a lack of glycosylation. 21269602 5 9 theme classical 948:956 arg1 cadherins 958:966 classical cadherins 948:966 classical cadherins 948:966 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 6 10 theme tryptophan 1036:1045 arg1 residues 1054:1061 two tryptophan anchor residues 1032:1061 two tryptophan anchor residues 1032:1061 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 6 11 theme I 1106:1106 arg1 cadherins 1108:1116 type I cadherins 1101:1116 type I cadherins 1101:1116 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 3 12 theme classical 549:557 arg1 cadherins 559:567 other classical cadherins 543:567 other classical cadherins 543:567 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 5 13 theme strand-swap 916:926 arg1 mechanism 928:936 the strand-swap mechanism 912:936 the strand-swap mechanism common to classical cadherins 912:966 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 4 14 theme protein 704:710 arg1 artifact 729:736 an artifact 726:736 an artifact that arises from a lack of glycosylation 726:777 Trimerization of the bacterially produced protein appears to be an artifact that arises from a lack of glycosylation. 21269602 4 14 theme protein 704:710 arg1 Trimerization 662:674 Trimerization 662:674 Trimerization of the bacterially produced protein 662:710 Trimerization of the bacterially produced protein appears to be an artifact that arises from a lack of glycosylation. 21269602 5 15 theme crystal 817:823 arg1 structure 825:833 the 2.1-Å-resolution crystal structure 796:833 the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins 796:966 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 6 16 theme adhesive 1072:1079 arg1 interface 1081:1089 the adhesive interface 1068:1089 the adhesive interface 1068:1089 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 4 17 theme produced 695:702 arg1 protein 704:710 the bacterially produced protein 679:710 the bacterially produced protein 679:710 Trimerization of the bacterially produced protein appears to be an artifact that arises from a lack of glycosylation. 21269602 5 18 theme VE-cadherin 842:852 arg1 region 869:874 the VE-cadherin EC1-2 adhesive region 838:874 the VE-cadherin EC1-2 adhesive region 838:874 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 5 19 theme 2.1-Å-resolution 800:815 arg1 structure 825:833 the 2.1-Å-resolution crystal structure 796:833 the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins 796:966 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 6 20 with common 972:977 arg1 cadherins 992:1000 type II cadherins 984:1000 type II cadherins 984:1000 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 3 21 theme trans 597:601 arg1 dimers 603:608 adhesive trans dimers 588:608 adhesive trans dimers 588:608 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 1 22 theme type 169:172 arg1 family 196:201 the type II classical cadherin family 165:201 the type II classical cadherin family of cell adhesion proteins 165:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 5 23 theme EC1-2 854:858 arg1 region 869:874 the VE-cadherin EC1-2 adhesive region 838:874 the VE-cadherin EC1-2 adhesive region 838:874 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 3 24 theme adhesive 588:595 arg1 dimers 603:608 adhesive trans dimers 588:608 adhesive trans dimers 588:608 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 7 25 with outlier 1216:1222 arg1 characteristics 1256:1270 characteristics 1256:1270 characteristics of both type I and type II subfamilies 1256:1309 Thus, VE-cadherin is an outlier among classical cadherins, with characteristics of both type I and type II subfamilies. 21269602 2 26 theme surface 386:392 arg1 trimers 394:400 cell surface trimers 381:400 cell surface trimers that bind between apposed cells to form hexamers 381:449 Previous investigations with a bacterially produced protein suggested that VE-cadherin forms cell surface trimers that bind between apposed cells to form hexamers. 21269602 5 27 theme adhesive 860:867 arg1 region 869:874 the VE-cadherin EC1-2 adhesive region 838:874 the VE-cadherin EC1-2 adhesive region 838:874 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 1 28 theme vascular 266:273 arg1 endothelium 275:285 the vascular endothelium 262:285 the vascular endothelium 262:285 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 29 theme Vascular 98:105 arg1 VE-cadherin 129:139 VE-cadherin 129:139 VE-cadherin 129:139 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 29 theme Vascular 98:105 arg1 cadherin 119:126 Vascular endothelial cadherin 98:126 Vascular endothelial cadherin (VE-cadherin) 98:140 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 29 theme Vascular 98:105 arg1 member 155:160 a divergent member 143:160 a divergent member of the type II classical cadherin family of cell adhesion proteins 143:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 30 theme classical 177:185 arg1 family 196:201 the type II classical cadherin family 165:201 the type II classical cadherin family of cell adhesion proteins 165:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 0 31 theme binding 14:20 arg1 mechanism 22:30 binding mechanism 14:30 binding mechanism 14:30 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 2 32 theme Previous 288:295 arg1 investigations 297:310 Previous investigations 288:310 Previous investigations with a bacterially produced protein 288:346 Previous investigations with a bacterially produced protein suggested that VE-cadherin forms cell surface trimers that bind between apposed cells to form hexamers. 21269602 1 33 theme cadherin 187:194 arg1 family 196:201 the type II classical cadherin family 165:201 the type II classical cadherin family of cell adhesion proteins 165:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 0 34 theme vascular 35:42 arg1 cadherin 56:63 vascular endothelial cadherin 35:63 vascular endothelial cadherin 35:63 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 6 35 theme strand-swap 1003:1013 arg1 binding 1015:1021 strand-swap binding 1003:1021 strand-swap binding 1003:1021 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 7 36 dep type 1280:1283 arg1 subfamilies 1299:1309 subfamilies 1299:1309 subfamilies 1299:1309 Thus, VE-cadherin is an outlier among classical cadherins, with characteristics of both type I and type II subfamilies. 21269602 2 37 theme apposed 420:426 arg1 cells 428:432 apposed cells 420:432 apposed cells 420:432 Previous investigations with a bacterially produced protein suggested that VE-cadherin forms cell surface trimers that bind between apposed cells to form hexamers. 21269602 3 38 theme located 627:633 arg1 monomers 618:625 monomers 618:625 monomers located on opposing cell surfaces 618:659 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 1 39 theme family 196:201 arg1 cadherin 119:126 Vascular endothelial cadherin 98:126 Vascular endothelial cadherin (VE-cadherin) 98:140 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 1 39 theme family 196:201 arg1 member 155:160 a divergent member 143:160 a divergent member of the type II classical cadherin family of cell adhesion proteins 143:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 6 40 theme type 984:987 arg1 cadherins 992:1000 type II cadherins 984:1000 type II cadherins 984:1000 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 0 41 theme cadherin 56:63 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 0 41 theme cadherin 56:63 arg1 mechanism 22:30 binding mechanism 14:30 binding mechanism 14:30 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 5 42 theme common 938:943 arg1 mechanism 928:936 the strand-swap mechanism 912:936 the strand-swap mechanism common to classical cadherins 912:966 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 6 43 theme hydrophobic 1171:1181 arg1 surface 1183:1189 a large nonswapped hydrophobic surface 1152:1189 a large nonswapped hydrophobic surface 1152:1189 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 3 44 theme ectodomains 509:519 arg1 studies 467:473 studies 467:473 studies of mammalian-produced VE-cadherin ectodomains 467:519 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 0 45 theme endothelial 44:54 arg1 cadherin 56:63 vascular endothelial cadherin 35:63 vascular endothelial cadherin 35:63 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 3 46 theme mammalian-produced 478:495 arg1 ectodomains 509:519 mammalian-produced VE-cadherin ectodomains 478:519 mammalian-produced VE-cadherin ectodomains 478:519 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 3 47 theme cell 647:650 arg1 surfaces 652:659 opposing cell surfaces 638:659 opposing cell surfaces 638:659 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 2 48 theme produced 331:338 arg1 protein 340:346 a bacterially produced protein 317:346 a bacterially produced protein 317:346 Previous investigations with a bacterially produced protein suggested that VE-cadherin forms cell surface trimers that bind between apposed cells to form hexamers. 21269602 6 49 theme nonswapped 1160:1169 arg1 surface 1183:1189 a large nonswapped hydrophobic surface 1152:1189 a large nonswapped hydrophobic surface 1152:1189 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 1 50 theme cell 206:209 arg1 proteins 220:227 cell adhesion proteins 206:227 cell adhesion proteins 206:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 5 51 theme region 869:874 arg1 structure 825:833 the 2.1-Å-resolution crystal structure 796:833 the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins 796:966 We also present the 2.1-Å-resolution crystal structure of the VE-cadherin EC1-2 adhesive region, which reveals homodimerization via the strand-swap mechanism common to classical cadherins. 21269602 3 52 theme VE-cadherin 497:507 arg1 ectodomains 509:519 mammalian-produced VE-cadherin ectodomains 478:519 mammalian-produced VE-cadherin ectodomains 478:519 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 1 53 theme adhesion 211:218 arg1 proteins 220:227 cell adhesion proteins 206:227 cell adhesion proteins 206:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 7 54 theme type 1280:1283 arg1 characteristics 1256:1270 characteristics 1256:1270 characteristics of both type I and type II subfamilies 1256:1309 Thus, VE-cadherin is an outlier among classical cadherins, with characteristics of both type I and type II subfamilies. 21269602 3 55 theme opposing 638:645 arg1 surfaces 652:659 opposing cell surfaces 638:659 opposing cell surfaces 638:659 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 2 56 theme cell 381:384 arg1 trimers 394:400 cell surface trimers 381:400 cell surface trimers that bind between apposed cells to form hexamers 381:449 Previous investigations with a bacterially produced protein suggested that VE-cadherin forms cell surface trimers that bind between apposed cells to form hexamers. 21269602 1 57 theme proteins 220:227 arg1 family 196:201 the type II classical cadherin family 165:201 the type II classical cadherin family of cell adhesion proteins 165:227 Vascular endothelial cadherin (VE-cadherin), a divergent member of the type II classical cadherin family of cell adhesion proteins, mediates homophilic adhesion in the vascular endothelium. 21269602 6 58 theme large 1154:1158 arg1 surface 1183:1189 a large nonswapped hydrophobic surface 1152:1189 a large nonswapped hydrophobic surface 1152:1189 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 21269602 0 59 dep Structure 0:8 arg1 cadherin 88:95 a divergent classical cadherin 66:95 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 0:96 Structure and binding mechanism of vascular endothelial cadherin: a divergent classical cadherin. 21269602 3 60 theme other 543:547 arg1 cadherins 559:567 other classical cadherins 543:567 other classical cadherins 543:567 Here we report studies of mammalian-produced VE-cadherin ectodomains suggesting that, like other classical cadherins, VE-cadherin forms adhesive trans dimers between monomers located on opposing cell surfaces. 21269602 7 61 theme classical 1230:1238 arg1 cadherins 1240:1248 classical cadherins 1230:1248 classical cadherins 1230:1248 Thus, VE-cadherin is an outlier among classical cadherins, with characteristics of both type I and type II subfamilies. 21269602 6 62 theme type 1101:1104 arg1 cadherins 1108:1116 type I cadherins 1101:1116 type I cadherins 1101:1116 In common with type II cadherins, strand-swap binding involves two tryptophan anchor residues, but the adhesive interface resembles type I cadherins in that VE-cadherin does not form a large nonswapped hydrophobic surface. 7827751 6 0 theme collagen 834:841 arg1 helix 850:854 the collagen triple helix 830:854 the collagen triple helix 830:854 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 4 1 theme Yaa 640:642 arg1 position 644:651 the Yaa position 636:651 the Yaa position 636:651 Partial or complete hydroxylation and glycosylation were found for all seven lysines in the Yaa position. 7827751 3 2 theme triplets 431:438 arg1 position 403:410 the Yaa position 395:410 the Yaa position of the Gly-Xaa-Yaa triplets 395:438 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 8 3 theme % 1263:1263 arg1 helix 1271:1275 11% alpha helix 1261:1275 11% alpha helix 1261:1275 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 0 4 theme XIV 72:74 arg1 XIV 72:74 human collagen XIV 57:74 human collagen XIV 57:74 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 0 4 theme XIV 72:74 arg1 domains 46:52 the triple-helical domains 27:52 the triple-helical domains of human collagen XIV 27:74 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 7 5 theme helix 999:1003 arg1 refolding 975:983 An almost perfect refolding 957:983 An almost perfect refolding of the triple helix 957:1003 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 9 6 theme intact 1341:1346 arg1 XIV 1357:1359 NC3 and intact collagen XIV 1333:1359 NC3 and intact collagen XIV 1333:1359 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 7 7 theme triple 992:997 arg1 helix 999:1003 the triple helix 988:1003 the triple helix 988:1003 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 9 8 theme degrees 1451:1457 arg1 C 1459:1459 44 degrees C 1448:1459 44 degrees C 1448:1459 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 9 9 theme melting 1425:1431 arg1 temperature 1433:1443 a melting temperature 1423:1443 a melting temperature of 44 degrees C 1423:1459 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 9 10 theme C 1459:1459 arg1 temperature 1433:1443 a melting temperature 1423:1443 a melting temperature of 44 degrees C 1423:1459 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 2 11 from residues 298:305 arg1 residues 328:335 the Col 1 and 149 residues 310:335 the Col 1 and 149 residues in the Col 2 domain 310:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 9 12 theme collagen 1348:1355 arg1 XIV 1357:1359 NC3 and intact collagen XIV 1333:1359 NC3 and intact collagen XIV 1333:1359 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 4 13 from lysines 625:631 arg1 position 644:651 the Yaa position 636:651 the Yaa position 636:651 Partial or complete hydroxylation and glycosylation were found for all seven lysines in the Yaa position. 7827751 8 14 theme dichroic 1174:1181 arg1 spectra 1183:1189 Circular dichroic spectra 1165:1189 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV 1165:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 2 15 theme Edman 251:255 arg1 degradation 257:267 Edman degradation 251:267 Edman degradation 251:267 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 1 16 theme nondenaturing 225:237 arg1 conditions 239:248 nondenaturing conditions 225:248 nondenaturing conditions 225:248 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 7 17 theme perfect 967:973 arg1 refolding 975:983 An almost perfect refolding 957:983 An almost perfect refolding of the triple helix 957:1003 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 3 18 theme Yaa 399:401 arg1 position 403:410 the Yaa position 395:410 the Yaa position of the Gly-Xaa-Yaa triplets 395:438 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 7 19 theme triple 1151:1156 arg1 helix 1158:1162 the triple helix 1147:1162 the triple helix 1147:1162 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 5 20 theme disulfide-linked 762:777 arg1 dimers 779:784 disulfide-linked dimers 762:784 disulfide-linked dimers 762:784 Domain Col 1 was joined by disulfide bonds into a trimer, while Col 2 appeared as a mixture of monomers and disulfide-linked dimers. 7827751 9 21 theme Comparative 1301:1311 arg1 profiles 1321:1328 Comparative melting profiles 1301:1328 Comparative melting profiles of NC3 and intact collagen XIV 1301:1359 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 2 22 theme Col 344:346 arg1 domain 350:355 the Col 2 domain 340:355 the Col 2 domain 340:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 9 23 contain have 1418:1421 arg2 temperature 1433:1443 a melting temperature 1423:1443 a melting temperature of 44 degrees C 1423:1459 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 9 23 contain have 1418:1421 arg1 helices 1387:1393 the triple helices 1376:1393 the triple helices in intact collagen XIV 1376:1416 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 6 24 theme dichroic 796:803 arg1 spectra 805:811 Circular dichroic spectra 787:811 Circular dichroic spectra 787:811 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 5 25 link disulfide-linked 762:777 arg1 dimers 779:784 disulfide-linked dimers 762:784 disulfide-linked dimers 762:784 Domain Col 1 was joined by disulfide bonds into a trimer, while Col 2 appeared as a mixture of monomers and disulfide-linked dimers. 7827751 5 26 theme disulfide 681:689 arg1 bonds 691:695 disulfide bonds 681:695 disulfide bonds 681:695 Domain Col 1 was joined by disulfide bonds into a trimer, while Col 2 appeared as a mixture of monomers and disulfide-linked dimers. 7827751 8 27 theme nontriple 1204:1212 arg1 XIV 1247:1249 collagen XIV 1238:1249 collagen XIV 1238:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 27 theme nontriple 1204:1212 arg1 NC3 1230:1232 NC3 1230:1232 NC3 1230:1232 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 27 theme nontriple 1204:1212 arg1 domain 1222:1227 the large nontriple helical domain 1194:1227 the large nontriple helical domain 1194:1227 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 1 28 theme pepsin 143:148 arg1 digest 150:155 a pepsin digest 141:155 a pepsin digest of human placental collagen XIV 141:187 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 2 29 from residues 328:335 arg1 domain 350:355 the Col 2 domain 340:355 the Col 2 domain 340:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 1 30 theme triple-helical 81:94 arg1 Col 105:107 Col 105:107 Col 105:107 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 1 30 theme triple-helical 81:94 arg1 Col 115:117 Col 115:117 Col 115:117 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 1 30 theme triple-helical 81:94 arg1 domains 96:102 Two triple-helical domains 77:102 Two triple-helical domains 77:102 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 9 31 theme triple 1380:1385 arg1 helices 1387:1393 the triple helices 1376:1393 the triple helices in intact collagen XIV 1376:1416 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 6 32 theme degrees 945:951 arg1 C 953:953 43 degrees C 942:953 43 degrees C 942:953 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 6 32 theme degrees 945:951 arg1 Col 935:937 Col 2 935:939 Col 2 (43 degrees C) 935:954 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 5 33 theme monomers 749:756 arg1 mixture 738:744 a mixture 736:744 a mixture of monomers and disulfide-linked dimers 736:784 Domain Col 1 was joined by disulfide bonds into a trimer, while Col 2 appeared as a mixture of monomers and disulfide-linked dimers. 7827751 8 34 theme domain 1222:1227 arg1 spectra 1183:1189 Circular dichroic spectra 1165:1189 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV 1165:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 9 35 theme intact 1398:1403 arg1 XIV 1414:1416 intact collagen XIV 1398:1416 intact collagen XIV 1398:1416 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 2 36 theme amino 286:290 arg1 acids 292:296 106 amino acids 282:296 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain 282:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 6 37 theme melting 885:891 arg1 temperatures 893:904 relatively high melting temperatures 869:904 relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C) 869:954 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 4 38 theme Partial 548:554 arg1 hydroxylation 568:580 hydroxylation 568:580 hydroxylation 568:580 Partial or complete hydroxylation and glycosylation were found for all seven lysines in the Yaa position. 7827751 6 39 theme Circular 787:794 arg1 spectra 805:811 Circular dichroic spectra 787:811 Circular dichroic spectra 787:811 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 8 40 theme helical 1214:1220 arg1 XIV 1247:1249 collagen XIV 1238:1249 collagen XIV 1238:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 40 theme helical 1214:1220 arg1 NC3 1230:1232 NC3 1230:1232 NC3 1230:1232 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 40 theme helical 1214:1220 arg1 domain 1222:1227 the large nontriple helical domain 1194:1227 the large nontriple helical domain 1194:1227 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 1 41 theme human 160:164 arg1 XIV 185:187 human placental collagen XIV 160:187 human placental collagen XIV 160:187 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 0 42 theme triple-helical 31:44 arg1 XIV 72:74 human collagen XIV 57:74 human collagen XIV 57:74 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 0 42 theme triple-helical 31:44 arg1 domains 46:52 the triple-helical domains 27:52 the triple-helical domains of human collagen XIV 27:74 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 6 43 theme high 880:883 arg1 temperatures 893:904 relatively high melting temperatures 869:904 relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C) 869:954 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 5 44 theme Domain 654:659 arg1 Col 661:663 Domain Col 1 654:665 Domain Col 1 654:665 Domain Col 1 was joined by disulfide bonds into a trimer, while Col 2 appeared as a mixture of monomers and disulfide-linked dimers. 7827751 3 45 theme prolines 383:390 arg1 one 369:371 one 369:371 one 369:371 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 3 45 theme prolines 383:390 arg1 prolines 383:390 the 37 prolines 376:390 the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets 376:438 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 3 46 theme triplet 532:538 arg1 repeat 540:545 the triplet repeat 528:545 the triplet repeat 528:545 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 7 47 theme helix 1158:1162 arg1 stability 1134:1142 the stability 1130:1142 the stability of the triple helix 1130:1162 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 7 47 theme helix 1158:1162 arg1 part 1125:1128 part 1125:1128 part 1125:1128 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 1 48 theme placental 166:174 arg1 XIV 185:187 human placental collagen XIV 160:187 human placental collagen XIV 160:187 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 9 49 from helices 1387:1393 arg1 XIV 1414:1416 intact collagen XIV 1398:1416 intact collagen XIV 1398:1416 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 9 50 theme collagen 1405:1412 arg1 XIV 1414:1416 intact collagen XIV 1398:1416 intact collagen XIV 1398:1416 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 3 51 from prolines 383:390 arg1 position 403:410 the Yaa position 395:410 the Yaa position of the Gly-Xaa-Yaa triplets 395:438 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 1 52 theme collagen 176:183 arg1 XIV 185:187 human placental collagen XIV 160:187 human placental collagen XIV 160:187 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 7 53 theme disulfide 1077:1085 arg1 bonds 1087:1091 disulfide bonds 1077:1091 disulfide bonds 1077:1091 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 8 54 theme large 1198:1202 arg1 XIV 1247:1249 collagen XIV 1238:1249 collagen XIV 1238:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 54 theme large 1198:1202 arg1 NC3 1230:1232 NC3 1230:1232 NC3 1230:1232 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 54 theme large 1198:1202 arg1 domain 1222:1227 the large nontriple helical domain 1194:1227 the large nontriple helical domain 1194:1227 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 2 55 theme Col 314:316 arg1 residues 328:335 the Col 1 and 149 residues 310:335 the Col 1 and 149 residues in the Col 2 domain 310:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 0 56 theme domains 46:52 arg1 stability 14:22 stability 14:22 stability 14:22 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 0 56 theme domains 46:52 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 8 57 theme % 1283:1283 arg1 structure 1290:1298 63% beta structure 1281:1298 63% beta structure 1281:1298 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 3 58 theme Gly-Xaa-Yaa 419:429 arg1 triplets 431:438 the Gly-Xaa-Yaa triplets 415:438 the Gly-Xaa-Yaa triplets 415:438 All except one of the 37 prolines in the Yaa position of the Gly-Xaa-Yaa triplets were completely hydroxylated to 4-hydroxyproline, and there were three imperfections in the triplet repeat. 7827751 9 59 theme XIV 1357:1359 arg1 profiles 1321:1328 Comparative melting profiles 1301:1328 Comparative melting profiles of NC3 and intact collagen XIV 1301:1359 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 8 60 theme 63 1281:1282 arg1 % 1283:1283 % 1283:1283 % 1283:1283 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 61 theme XIV 1247:1249 arg1 XIV 1247:1249 collagen XIV 1238:1249 collagen XIV 1238:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 61 theme XIV 1247:1249 arg1 NC3 1230:1232 NC3 1230:1232 NC3 1230:1232 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 61 theme XIV 1247:1249 arg1 domain 1222:1227 the large nontriple helical domain 1194:1227 the large nontriple helical domain 1194:1227 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 9 62 theme NC3 1333:1335 arg1 XIV 1357:1359 NC3 and intact collagen XIV 1333:1359 NC3 and intact collagen XIV 1333:1359 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 2 63 dep acids 292:296 arg1 residues 298:305 residues 298:305 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain 282:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 1 64 theme XIV 185:187 arg1 digest 150:155 a pepsin digest 141:155 a pepsin digest of human placental collagen XIV 141:187 Two triple-helical domains, Col 1 and Col 2, were obtained from a pepsin digest of human placental collagen XIV and separated from each other under nondenaturing conditions. 7827751 8 65 theme alpha 1265:1269 arg1 helix 1271:1275 11% alpha helix 1261:1275 11% alpha helix 1261:1275 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 66 theme collagen 1238:1245 arg1 XIV 1247:1249 collagen XIV 1238:1249 collagen XIV 1238:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 7 67 theme folding 1101:1107 arg1 kinetics 1109:1116 the folding kinetics 1097:1116 the folding kinetics 1097:1116 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 8 68 theme Circular 1165:1172 arg1 spectra 1183:1189 Circular dichroic spectra 1165:1189 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV 1165:1249 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 9 69 theme melting 1313:1319 arg1 profiles 1321:1328 Comparative melting profiles 1301:1328 Comparative melting profiles of NC3 and intact collagen XIV 1301:1359 Comparative melting profiles of NC3 and intact collagen XIV indicated that the triple helices in intact collagen XIV have a melting temperature of 44 degrees C. 7827751 0 70 theme collagen 63:70 arg1 XIV 72:74 human collagen XIV 57:74 human collagen XIV 57:74 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 8 71 theme beta 1285:1288 arg1 structure 1290:1298 63% beta structure 1281:1298 63% beta structure 1281:1298 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 8 72 theme 11 1261:1262 arg1 % 1263:1263 % 1263:1263 % 1263:1263 Circular dichroic spectra of the large nontriple helical domain, NC3, of collagen XIV indicated 11% alpha helix and 63% beta structure. 7827751 5 73 theme dimers 779:784 arg1 mixture 738:744 a mixture 736:744 a mixture of monomers and disulfide-linked dimers 736:784 Domain Col 1 was joined by disulfide bonds into a trimer, while Col 2 appeared as a mixture of monomers and disulfide-linked dimers. 7827751 4 74 theme complete 559:566 arg1 hydroxylation 568:580 hydroxylation 568:580 hydroxylation 568:580 Partial or complete hydroxylation and glycosylation were found for all seven lysines in the Yaa position. 7827751 0 75 theme human 57:61 arg1 XIV 72:74 human collagen XIV 57:74 human collagen XIV 57:74 Structure and stability of the triple-helical domains of human collagen XIV. 7827751 6 76 theme triple 843:848 arg1 helix 850:854 the collagen triple helix 830:854 the collagen triple helix 830:854 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 2 77 theme 149 324:326 arg1 residues 328:335 the Col 1 and 149 residues 310:335 the Col 1 and 149 residues in the Col 2 domain 310:355 Edman degradation demonstrated 106 amino acids residues in the Col 1 and 149 residues in the Col 2 domain. 7827751 7 78 theme bonds 1087:1091 arg1 importance 1063:1072 the importance 1059:1072 the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix 1059:1162 An almost perfect refolding of the triple helix was observed for Col 1 but not for Col 2, emphasizing the importance of disulfide bonds for the folding kinetics and in part the stability of the triple helix. 7827751 6 79 theme degrees 920:926 arg1 C 928:928 38 degrees C 917:928 38 degrees C 917:928 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 7827751 6 79 theme degrees 920:926 arg1 Col 910:912 Col 1 910:914 Col 1 (38 degrees C) 910:929 Circular dichroic spectra were typical for the collagen triple helix and revealed relatively high melting temperatures for Col 1 (38 degrees C) and Col 2 (43 degrees C). 3524673 9 0 theme factor 792:797 arg1 sequence 765:772 The sequence 761:772 The sequence of von Willebrand factor 761:797 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 11 1 theme half-cystinyl 1120:1132 arg1 residues 1134:1141 half-cystinyl residues 1120:1141 half-cystinyl residues 1120:1141 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 8 2 theme sequence 694:701 arg1 Asn-Ser-Cys 703:713 the sequence Asn-Ser-Cys 690:713 the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 690:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 11 3 contain contain 1100:1106 arg1 regions 1076:1082 carboxyl-terminal regions 1058:1082 carboxyl-terminal regions 1058:1082 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 11 3 contain contain 1100:1106 arg1 amino- 1047:1052 amino- 1047:1052 amino- 1047:1052 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 11 3 contain contain 1100:1106 arg2 clusters 1108:1115 clusters 1108:1115 clusters of half-cystinyl residues 1108:1141 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 7 4 theme Thr/Ser-linked 589:602 arg1 chains 620:625 10 Thr/Ser-linked oligosaccharide chains 586:625 10 Thr/Ser-linked oligosaccharide chains 586:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 9 5 theme several 808:814 arg1 regions 816:822 several regions 808:822 several regions bearing evidence of internal gene duplication of ancestral sequences 808:891 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 11 6 theme carboxyl-terminal 1058:1074 arg1 regions 1076:1082 carboxyl-terminal regions 1058:1082 carboxyl-terminal regions 1058:1082 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 3 7 theme nucleotide 329:338 arg1 sequence 340:347 the nucleotide sequence 325:347 the nucleotide sequence of a cDNA clone [Sadler 325:371 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 3 7 theme nucleotide 329:338 arg1 Shelton-Inloes 380:393 Shelton-Inloes 380:393 Shelton-Inloes 380:393 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 11 8 theme molecule 1091:1098 arg1 amino- 1047:1052 amino- 1047:1052 amino- 1047:1052 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 11 8 theme molecule 1091:1098 arg1 regions 1076:1082 carboxyl-terminal regions 1058:1082 carboxyl-terminal regions 1058:1082 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 0 9 theme Amino 0:4 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of human von Willebrand factor 0:49 Amino acid sequence of human von Willebrand factor. 3524673 8 10 theme usual 731:735 arg1 sequence 751:758 the usual Asn-X-Ser/Thr sequence 727:758 the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 690:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 10 11 from residues 966:973 arg1 site 1018:1021 a cell attachment site 1000:1021 a cell attachment site 1000:1021 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 11 from residues 966:973 arg1 Arg-Gly-Asp-Ser 946:960 the tetrapeptide sequence Arg-Gly-Asp-Ser 920:960 the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747) 920:984 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 7 12 link Thr/Ser-linked 589:602 arg1 chains 620:625 10 Thr/Ser-linked oligosaccharide chains 586:625 10 Thr/Ser-linked oligosaccharide chains 586:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 1 13 theme complete 56:63 arg1 sequence 76:83 The complete amino acid sequence 52:83 The complete amino acid sequence of human von Willebrand factor (vWF) 52:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 3 14 theme [Sadler 365:371 arg1 sequence 340:347 the nucleotide sequence 325:347 the nucleotide sequence of a cDNA clone [Sadler 325:371 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 3 14 theme [Sadler 365:371 arg1 Shelton-Inloes 380:393 Shelton-Inloes 380:393 Shelton-Inloes 380:393 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 10 15 from site 1018:1021 arg1 fibronectin 1030:1040 fibronectin 1030:1040 fibronectin 1030:1040 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 1 16 theme amino 65:69 arg1 sequence 76:83 The complete amino acid sequence 52:83 The complete amino acid sequence of human von Willebrand factor (vWF) 52:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 0 17 theme acid 6:9 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of human von Willebrand factor 0:49 Amino acid sequence of human von Willebrand factor. 3524673 8 18 attach linked 662:667 arg2 One 628:630 One 628:630 One 628:630 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 8 18 attach linked 662:667 arg2 chains 652:657 the carbohydrate chains 635:657 the carbohydrate chains 635:657 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 8 18 attach linked 662:667 arg1 residue 679:685 an Asn residue 672:685 an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 672:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 1 19 theme acid 71:74 arg1 sequence 76:83 The complete amino acid sequence 52:83 The complete amino acid sequence of human von Willebrand factor (vWF) 52:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 8 20 theme Asn-X-Ser/Thr 737:749 arg1 sequence 751:758 the usual Asn-X-Ser/Thr sequence 727:758 the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 690:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 3 21 theme cDNA 354:357 arg1 [Sadler 365:371 a cDNA clone [Sadler 352:371 a cDNA clone [Sadler 352:371 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 10 22 theme attachment 1007:1016 arg1 site 1018:1021 a cell attachment site 1000:1021 a cell attachment site 1000:1021 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 22 theme attachment 1007:1016 arg1 Arg-Gly-Asp-Ser 946:960 the tetrapeptide sequence Arg-Gly-Asp-Ser 920:960 the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747) 920:984 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 0 23 theme Willebrand 33:42 arg1 factor 44:49 human von Willebrand factor 23:49 human von Willebrand factor 23:49 Amino acid sequence of human von Willebrand factor. 3524673 9 24 theme gene 853:856 arg1 duplication 858:868 internal gene duplication 844:868 internal gene duplication of ancestral sequences 844:891 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 10 25 theme cell 1002:1005 arg1 site 1018:1021 a cell attachment site 1000:1021 a cell attachment site 1000:1021 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 25 theme cell 1002:1005 arg1 Arg-Gly-Asp-Ser 946:960 the tetrapeptide sequence Arg-Gly-Asp-Ser 920:960 the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747) 920:984 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 12 26 theme complement 1201:1210 arg1 B 1219:1219 human complement factor B 1195:1219 human complement factor B 1195:1219 The sequence is unique except for some homology to human complement factor B. 3524673 9 27 theme duplication 858:868 arg1 evidence 832:839 evidence 832:839 evidence of internal gene duplication of ancestral sequences 832:891 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 8 28 from residue 679:685 arg1 Asn-Ser-Cys 703:713 the sequence Asn-Ser-Cys 690:713 the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 690:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 8 28 from residue 679:685 arg1 sequence 751:758 the usual Asn-X-Ser/Thr sequence 727:758 the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 690:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 1 29 theme human 88:92 arg1 factor 109:114 human von Willebrand factor 88:114 human von Willebrand factor (vWF) 88:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 1 29 theme human 88:92 arg1 vWF 117:119 vWF 117:119 vWF 117:119 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 0 30 theme von 29:31 arg1 factor 44:49 human von Willebrand factor 23:49 human von Willebrand factor 23:49 Amino acid sequence of human von Willebrand factor. 3524673 2 31 theme S-carboxymethylated 191:209 arg1 protein 211:217 the S-carboxymethylated protein 187:217 the S-carboxymethylated protein 187:217 Most of the sequence was determined by analysis of the S-carboxymethylated protein. 3524673 0 32 theme human 23:27 arg1 factor 44:49 human von Willebrand factor 23:49 human von Willebrand factor 23:49 Amino acid sequence of human von Willebrand factor. 3524673 9 33 theme ancestral 873:881 arg1 sequences 883:891 ancestral sequences 873:891 ancestral sequences 873:891 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 7 34 theme acid 543:546 arg1 residues 548:555 2050 amino acid residues 532:555 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains 532:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 8 35 theme chains 652:657 arg1 chains 652:657 the carbohydrate chains 635:657 the carbohydrate chains 635:657 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 8 35 theme chains 652:657 arg1 One 628:630 One 628:630 One 628:630 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 7 36 contain containing 557:566 arg1 residues 548:555 2050 amino acid residues 532:555 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains 532:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 7 36 contain containing 557:566 arg2 chains 620:625 10 Thr/Ser-linked oligosaccharide chains 586:625 10 Thr/Ser-linked oligosaccharide chains 586:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 7 36 contain containing 557:566 arg2 Asn-linked 571:580 Asn-linked 571:580 Asn-linked 571:580 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 0 37 theme factor 44:49 arg1 sequence 11:18 Amino acid sequence 0:18 Amino acid sequence of human von Willebrand factor 0:49 Amino acid sequence of human von Willebrand factor. 3524673 11 38 theme residues 1134:1141 arg1 clusters 1108:1115 clusters 1108:1115 clusters of half-cystinyl residues 1108:1141 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 9 39 theme Willebrand 781:790 arg1 factor 792:797 von Willebrand factor 777:797 von Willebrand factor 777:797 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 1 40 theme von 94:96 arg1 factor 109:114 human von Willebrand factor 88:114 human von Willebrand factor (vWF) 88:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 1 40 theme von 94:96 arg1 vWF 117:119 vWF 117:119 vWF 117:119 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 3 41 dep overlaps 225:232 arg1 obtained 285:292 obtained 285:292 were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc 280:462 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 3 41 dep overlaps 225:232 arg1 provided 238:245 provided 238:245 overlaps not provided by the protein sequence analysis 225:278 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 9 42 theme sequences 883:891 arg1 duplication 858:868 internal gene duplication 844:868 internal gene duplication of ancestral sequences 844:891 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 10 43 theme sequence 937:944 arg1 site 1018:1021 a cell attachment site 1000:1021 a cell attachment site 1000:1021 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 43 theme sequence 937:944 arg1 Arg-Gly-Asp-Ser 946:960 the tetrapeptide sequence Arg-Gly-Asp-Ser 920:960 the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747) 920:984 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 1 44 theme Willebrand 98:107 arg1 factor 109:114 human von Willebrand factor 88:114 human von Willebrand factor (vWF) 88:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 1 44 theme Willebrand 98:107 arg1 vWF 117:119 vWF 117:119 vWF 117:119 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 10 45 dep residues 966:973 arg1 1744-1747 975:983 1744-1747 975:983 1744-1747 975:983 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 3 46 theme protein 254:260 arg1 analysis 271:278 the protein sequence analysis 250:278 the protein sequence analysis 250:278 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 7 47 theme amino 537:541 arg1 residues 548:555 2050 amino acid residues 532:555 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains 532:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 12 48 theme human 1195:1199 arg1 B 1219:1219 human complement factor B 1195:1219 human complement factor B 1195:1219 The sequence is unique except for some homology to human complement factor B. 3524673 1 49 theme factor 109:114 arg1 sequence 76:83 The complete amino acid sequence 52:83 The complete amino acid sequence of human von Willebrand factor (vWF) 52:120 The complete amino acid sequence of human von Willebrand factor (vWF) is presented. 3524673 8 50 theme Asn 675:677 arg1 residue 679:685 an Asn residue 672:685 an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence 672:758 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 3524673 3 51 theme sequence 262:269 arg1 analysis 271:278 the protein sequence analysis 250:278 the protein sequence analysis 250:278 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 10 52 theme tetrapeptide 924:935 arg1 site 1018:1021 a cell attachment site 1000:1021 a cell attachment site 1000:1021 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 52 theme tetrapeptide 924:935 arg1 Arg-Gly-Asp-Ser 946:960 the tetrapeptide sequence Arg-Gly-Asp-Ser 920:960 the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747) 920:984 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 11 53 dep amino- 1047:1052 arg1 The 1043:1045 The 1043:1045 The 1043:1045 The amino- and carboxyl-terminal regions of the molecule contain clusters of half-cystinyl residues. 3524673 9 54 theme von 777:779 arg1 factor 792:797 von Willebrand factor 777:797 von Willebrand factor 777:797 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 12 55 theme factor 1212:1217 arg1 B 1219:1219 human complement factor B 1195:1219 human complement factor B 1195:1219 The sequence is unique except for some homology to human complement factor B. 3524673 9 56 theme internal 844:851 arg1 duplication 858:868 internal gene duplication 844:868 internal gene duplication of ancestral sequences 844:891 The sequence of von Willebrand factor includes several regions bearing evidence of internal gene duplication of ancestral sequences. 3524673 10 57 contain contains 911:918 arg2 Arg-Gly-Asp-Ser 946:960 the tetrapeptide sequence Arg-Gly-Asp-Ser 920:960 the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747) 920:984 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 57 contain contains 911:918 arg1 protein 898:904 The protein 894:904 The protein 894:904 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 10 57 contain contains 911:918 arg2 site 1018:1021 a cell attachment site 1000:1021 a cell attachment site 1000:1021 The protein also contains the tetrapeptide sequence Arg-Gly-Asp-Ser (at residues 1744-1747), which may be a cell attachment site, as in fibronectin. 3524673 7 58 theme oligosaccharide 604:618 arg1 chains 620:625 10 Thr/Ser-linked oligosaccharide chains 586:625 10 Thr/Ser-linked oligosaccharide chains 586:625 The protein is composed of 2050 amino acid residues containing 12 Asn-linked and 10 Thr/Ser-linked oligosaccharide chains. 3524673 3 59 theme clone 359:363 arg1 [Sadler 365:371 a cDNA clone [Sadler 352:371 a cDNA clone [Sadler 352:371 Some overlaps not provided by the protein sequence analysis were obtained from the sequence predicted by the nucleotide sequence of a cDNA clone [Sadler, J.E., Shelton-Inloes, B.B., Sorace, J., Harlan, M., Titani, K., & Davie, E.W. (1985) Proc. 3524673 2 60 theme protein 211:217 arg1 analysis 175:182 analysis 175:182 analysis of the S-carboxymethylated protein 175:217 Most of the sequence was determined by analysis of the S-carboxymethylated protein. 3524673 8 61 theme carbohydrate 639:650 arg1 chains 652:657 the carbohydrate chains 635:657 the carbohydrate chains 635:657 One of the carbohydrate chains is linked to an Asn residue in the sequence Asn-Ser-Cys rather than the usual Asn-X-Ser/Thr sequence. 12888274 7 0 theme metal 1004:1008 arg1 ion 1010:1012 The metal ion 1000:1012 The metal ion present in the active site of arylsulfatase A isolated from human placenta 1000:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 0 theme metal 1004:1008 arg1 Ca 1092:1093 Ca 1092:1093 Ca(2+) 1092:1097 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 3 1 theme residue 504:510 arg1 Cbeta 495:499 the Cbeta 491:499 the Cbeta of residue 69 491:513 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 7 2 attach present 1014:1020 arg1 A 1058:1058 arylsulfatase A 1044:1058 arylsulfatase A isolated from human placenta 1044:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 2 attach present 1014:1020 arg2 Ca 1092:1093 Ca 1092:1093 Ca(2+) 1092:1097 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 2 attach present 1014:1020 arg1 site 1036:1039 the active site 1025:1039 the active site of arylsulfatase A isolated from human placenta 1025:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 2 attach present 1014:1020 arg2 ion 1010:1012 The metal ion 1000:1012 The metal ion present in the active site of arylsulfatase A isolated from human placenta 1000:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 0 3 theme A 79:79 arg1 intermediate 32:43 a covalent intermediate 21:43 a covalent intermediate of endogenous human arylsulfatase A 21:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 3 4 attach present 480:486 arg1 Cbeta 495:499 the Cbeta 491:499 the Cbeta of residue 69 491:513 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 3 4 attach present 480:486 arg2 present 480:486 present 480:486 present 480:486 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 3 4 attach present 480:486 arg2 group 471:475 A hydroxyl group 460:475 A hydroxyl group 460:475 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 1 5 theme 4-methylumbelliferyl 162:181 arg1 phosphate 183:191 4-methylumbelliferyl phosphate 162:191 4-methylumbelliferyl phosphate 162:191 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 7 6 theme recombinant 1151:1161 arg1 enzyme 1163:1168 the recombinant enzyme 1147:1168 the recombinant enzyme 1147:1168 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 6 7 theme phosphate 911:919 arg1 esters 921:926 phosphate esters 911:926 phosphate esters 911:926 The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. 12888274 7 8 theme enzyme 1163:1168 arg1 structure 1134:1142 the structure 1130:1142 the structure of the recombinant enzyme 1130:1168 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 4 9 from importance 627:636 arg1 mechanism 700:708 the arylsulfatase's catalytic mechanism 670:708 the arylsulfatase's catalytic mechanism 670:708 The structures confirm the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism. 12888274 7 10 attach isolated 1060:1067 arg1 placenta 1080:1087 human placenta 1074:1087 human placenta 1074:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 10 attach isolated 1060:1067 arg2 A 1058:1058 arylsulfatase A 1044:1058 arylsulfatase A isolated from human placenta 1044:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 4 11 theme catalytic 690:698 arg1 mechanism 700:708 the arylsulfatase's catalytic mechanism 670:708 the arylsulfatase's catalytic mechanism 670:708 The structures confirm the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism. 12888274 1 12 contain containing 151:160 arg1 solutions 141:149 solutions 141:149 solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine 141:217 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 1 12 contain containing 151:160 arg2 O-phospho-DL-tyrosine 197:217 O-phospho-DL-tyrosine 197:217 O-phospho-DL-tyrosine 197:217 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 1 12 contain containing 151:160 arg2 phosphate 183:191 4-methylumbelliferyl phosphate 162:191 4-methylumbelliferyl phosphate 162:191 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 4 13 theme intermediate 654:665 arg1 importance 627:636 the importance 623:636 the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism 623:708 The structures confirm the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism. 12888274 5 14 theme covalent 748:755 arg1 bond 757:760 an apparently stable covalent bond 727:760 an apparently stable covalent bond 727:760 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 6 15 theme phosphate 892:900 arg1 ions 902:905 phosphate ions 892:905 phosphate ions 892:905 The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. 12888274 4 16 theme gem-diol 645:652 arg1 intermediate 654:665 the gem-diol intermediate 641:665 the gem-diol intermediate 641:665 The structures confirm the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism. 12888274 5 17 theme bond 757:760 arg1 consistent 765:774 consistent 765:774 consistent 765:774 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 5 17 theme bond 757:760 arg1 presence 715:722 The presence 711:722 The presence of an apparently stable covalent bond 711:760 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 3 18 theme possible 551:558 arg1 forms 572:576 one out of two possible stereomeric forms 536:576 one out of two possible stereomeric forms 536:576 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 0 19 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A 0:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 3 20 from Cbeta 495:499 arg1 group 471:475 A hydroxyl group 460:475 A hydroxyl group 460:475 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 3 20 from Cbeta 495:499 arg1 present 480:486 present 480:486 present 480:486 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 2 21 theme crucial 328:334 arg1 residue 320:326 a residue 318:326 a residue crucial for catalytic activity 318:357 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 2 21 theme crucial 328:334 arg1 position 305:312 position 69 305:315 position 69 305:315 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 7 22 from present 1014:1020 arg1 A 1058:1058 arylsulfatase A 1044:1058 arylsulfatase A isolated from human placenta 1044:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 22 from present 1014:1020 arg1 site 1036:1039 the active site 1025:1039 the active site of arylsulfatase A isolated from human placenta 1025:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 5 23 theme human 824:828 arg1 A 844:844 human arylsulfatase A 824:844 human arylsulfatase A 824:844 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 5 24 theme phosphatase 790:800 arg1 activity 802:809 the weak phosphatase activity 781:809 the weak phosphatase activity observed for human arylsulfatase A 781:844 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 3 25 located present 480:486 arg1 Cbeta 495:499 the Cbeta 491:499 the Cbeta of residue 69 491:513 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 3 25 located present 480:486 arg2 present 480:486 present 480:486 present 480:486 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 3 25 located present 480:486 arg2 group 471:475 A hydroxyl group 460:475 A hydroxyl group 460:475 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 2 26 theme phosphate 442:450 arg1 moiety 452:457 the phosphate moiety 438:457 the phosphate moiety 438:457 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 1 27 theme human 100:104 arg1 A 120:120 human arylsulfatase A 100:120 human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine 100:217 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 2 28 from formylglycine 288:300 arg1 residue 320:326 a residue 318:326 a residue crucial for catalytic activity 318:357 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 2 28 from formylglycine 288:300 arg1 position 305:312 position 69 305:315 position 69 305:315 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 0 29 theme intermediate 32:43 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A 0:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 6 30 theme complexes 869:877 arg1 structures 851:860 The structures 847:860 The structures of the complexes 847:877 The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. 12888274 3 31 theme hydroxyl 462:469 arg1 group 471:475 A hydroxyl group 460:475 A hydroxyl group 460:475 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 3 31 theme hydroxyl 462:469 arg1 present 480:486 present 480:486 present 480:486 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 0 32 theme covalent 23:30 arg1 intermediate 32:43 a covalent intermediate 21:43 a covalent intermediate of endogenous human arylsulfatase A 21:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 6 33 theme covalent 970:977 arg1 modes 979:983 non-covalent and covalent modes 953:983 non-covalent and covalent modes 953:983 The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. 12888274 5 34 theme weak 785:788 arg1 activity 802:809 the weak phosphatase activity 781:809 the weak phosphatase activity observed for human arylsulfatase A 781:844 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 7 35 theme active 1029:1034 arg1 A 1058:1058 arylsulfatase A 1044:1058 arylsulfatase A isolated from human placenta 1044:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 35 theme active 1029:1034 arg1 site 1036:1039 the active site 1025:1039 the active site of arylsulfatase A isolated from human placenta 1025:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 1 36 theme 2.7- 243:246 arg1 resolution 258:267 2.7- and 3.2-A resolution 243:267 2.7- and 3.2-A resolution 243:267 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 0 37 theme endogenous 48:57 arg1 A 79:79 endogenous human arylsulfatase A 48:79 endogenous human arylsulfatase A 48:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 7 38 theme A 1058:1058 arg1 A 1058:1058 arylsulfatase A 1044:1058 arylsulfatase A isolated from human placenta 1044:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 38 theme A 1058:1058 arg1 site 1036:1039 the active site 1025:1039 the active site of arylsulfatase A isolated from human placenta 1025:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 3 39 theme forms 572:576 arg1 formation 523:531 the formation 519:531 the formation of one out of two possible stereomeric forms 519:576 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12888274 1 40 theme arylsulfatase 106:118 arg1 A 120:120 human arylsulfatase A 100:120 human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine 100:217 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 7 41 theme human 1074:1078 arg1 placenta 1080:1087 human placenta 1074:1087 human placenta 1074:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 6 42 theme non-covalent 953:964 arg1 modes 979:983 non-covalent and covalent modes 953:983 non-covalent and covalent modes 953:983 The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. 12888274 1 43 theme A 120:120 arg1 crystals 122:129 human arylsulfatase A crystals 100:129 human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine 100:217 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 5 44 with consistent 765:774 arg1 activity 802:809 the weak phosphatase activity 781:809 the weak phosphatase activity observed for human arylsulfatase A 781:844 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 1 45 theme crystals 122:129 arg1 structures 86:95 The structures 82:95 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine 82:217 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 5 46 theme stable 741:746 arg1 bond 757:760 an apparently stable covalent bond 727:760 an apparently stable covalent bond 727:760 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 7 47 theme arylsulfatase 1044:1056 arg1 A 1058:1058 arylsulfatase A 1044:1058 arylsulfatase A isolated from human placenta 1044:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 2 48 theme catalytic 340:348 arg1 activity 350:357 catalytic activity 340:357 catalytic activity 340:357 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 1 49 theme 3.2-A 252:256 arg1 resolution 258:267 2.7- and 3.2-A resolution 243:267 2.7- and 3.2-A resolution 243:267 The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. 12888274 0 50 theme arylsulfatase 65:77 arg1 A 79:79 endogenous human arylsulfatase A 48:79 endogenous human arylsulfatase A 48:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 2 51 theme covalent 421:428 arg1 bond 430:433 a covalent bond 419:433 a covalent bond to the phosphate moiety 419:457 The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. 12888274 7 52 theme present 1014:1020 arg1 ion 1010:1012 The metal ion 1000:1012 The metal ion present in the active site of arylsulfatase A isolated from human placenta 1000:1087 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 7 52 theme present 1014:1020 arg1 Ca 1092:1093 Ca 1092:1093 Ca(2+) 1092:1097 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 0 53 theme human 59:63 arg1 A 79:79 endogenous human arylsulfatase A 48:79 endogenous human arylsulfatase A 48:79 Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. 12888274 7 54 from site 1036:1039 arg1 present 1014:1020 present 1014:1020 present 1014:1020 The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. 12888274 5 55 theme arylsulfatase 830:842 arg1 A 844:844 human arylsulfatase A 824:844 human arylsulfatase A 824:844 The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. 12888274 3 56 theme stereomeric 560:570 arg1 forms 572:576 one out of two possible stereomeric forms 536:576 one out of two possible stereomeric forms 536:576 A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. 12657638 2 0 theme x-ray 418:422 arg1 crystallography 424:438 x-ray crystallography 418:438 x-ray crystallography 418:438 The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. 12657638 5 1 from role 1006:1009 arg1 catalysis 1035:1043 catalysis 1035:1043 catalysis 1035:1043 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 1 2 theme tumor 215:219 arg1 cells 221:225 breast tumor cells 208:225 breast tumor cells 208:225 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 0 3 theme association 75:85 arg1 roles 57:61 functional roles 46:61 functional roles of membrane association 46:85 Structure of human estrone sulfatase suggests functional roles of membrane association. 12657638 5 4 theme " 931:931 arg1 cavity 912:917 a cavity 910:917 a cavity of the "gill" of the "mushroom 910:948 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 4 5 theme hydrophobic 633:643 arg1 each 654:657 each 654:657 each 654:657 These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. 12657638 4 5 theme hydrophobic 633:643 arg1 helices 645:651 These highly hydrophobic helices 620:651 These highly hydrophobic helices 620:651 These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. 12657638 4 6 theme ER 799:800 arg1 lumen 802:806 the ER lumen 795:806 the ER lumen 795:806 These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. 12657638 6 7 theme ER 1108:1109 arg1 membrane 1111:1118 the ER membrane 1104:1118 the ER membrane in dictating the form 1104:1140 This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes. 12657638 5 8 from deep 902:905 arg1 cavity 912:917 a cavity 910:917 a cavity of the "gill" of the "mushroom 910:948 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 5 9 theme active 882:887 arg1 site 889:892 the active site 878:892 the active site 878:892 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 5 10 theme domain 843:848 arg1 location 813:820 The location 809:820 The location of the transmembrane domain 809:848 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 5 10 theme domain 843:848 arg1 such 853:856 such 853:856 such 853:856 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 1 11 theme key 140:142 arg1 enzymes 144:150 the key enzymes 136:150 the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells 136:225 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 12 theme Estrone 88:94 arg1 enzymes 144:150 the key enzymes 136:150 the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells 136:225 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 12 theme Estrone 88:94 arg1 sulfatase 96:104 Estrone sulfatase 88:104 Estrone sulfatase (ES; 562 amino acids) 88:126 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 12 theme Estrone 88:94 arg1 one 129:131 one 129:131 one 129:131 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 12 theme Estrone 88:94 arg1 ES 107:108 ES 107:108 ES 107:108 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 13 theme enzymes 144:150 arg1 enzymes 144:150 the key enzymes 136:150 the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells 136:225 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 13 theme enzymes 144:150 arg1 sulfatase 96:104 Estrone sulfatase 88:104 Estrone sulfatase (ES; 562 amino acids) 88:126 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 13 theme enzymes 144:150 arg1 one 129:131 one 129:131 one 129:131 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 4 14 theme membrane 771:778 arg1 surface 780:786 the membrane surface 767:786 the membrane surface facing the ER lumen 767:806 These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. 12657638 1 15 theme responsible 152:162 arg1 enzymes 144:150 the key enzymes 136:150 the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells 136:225 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 0 16 theme estrone 19:25 arg1 sulfatase 27:35 human estrone sulfatase 13:35 human estrone sulfatase 13:35 Structure of human estrone sulfatase suggests functional roles of membrane association. 12657638 5 17 theme lipid 1018:1022 arg1 bilayer 1024:1030 the lipid bilayer 1014:1030 the lipid bilayer 1014:1030 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 6 18 theme enzymes 1174:1180 arg1 membrane 1111:1118 the ER membrane 1104:1118 the ER membrane in dictating the form 1104:1140 This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes. 12657638 6 18 theme enzymes 1174:1180 arg1 function 1150:1157 the function 1146:1157 the function of ER-resident enzymes 1146:1180 This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes. 12657638 3 19 theme antiparallel 489:500 arg1 alpha-helices 502:514 two antiparallel alpha-helices 485:514 two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape 485:617 This structure shows a domain consisting of two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape. 12657638 2 20 theme human 357:361 arg1 placentas 363:371 human placentas 357:371 human placentas 357:371 The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. 12657638 2 21 theme placentas 363:371 arg1 fraction 345:352 the microsomal fraction 330:352 the microsomal fraction of human placentas 330:371 The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. 12657638 5 22 theme bilayer 1024:1030 arg1 role 1006:1009 a role 1004:1009 a role of the lipid bilayer in catalysis 1004:1043 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 6 23 theme ER-resident 1162:1172 arg1 enzymes 1174:1180 ER-resident enzymes 1162:1180 ER-resident enzymes 1162:1180 This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes. 12657638 1 24 theme high 180:183 arg1 levels 185:190 high levels 180:190 high levels of estrogens 180:203 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 0 25 theme sulfatase 27:35 arg1 Structure 0:8 Structure 0:8 Structure of human estrone sulfatase 0:35 Structure of human estrone sulfatase suggests functional roles of membrane association. 12657638 2 26 theme 2.60-A 397:402 arg1 resolution 404:413 2.60-A resolution 397:413 2.60-A resolution 397:413 The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. 12657638 5 27 theme mushroom 941:948 arg1 " 931:931 the "gill" 922:931 the "gill" of the "mushroom 922:948 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 3 28 theme spherical 547:555 arg1 molecule 557:564 the roughly spherical molecule 535:564 the roughly spherical molecule 535:564 This structure shows a domain consisting of two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape. 12657638 4 29 from domain 757:762 arg1 surface 780:786 the membrane surface 767:786 the membrane surface facing the ER lumen 767:806 These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. 12657638 2 30 theme microsomal 334:343 arg1 fraction 345:352 the microsomal fraction 330:352 the microsomal fraction of human placentas 330:371 The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. 12657638 3 31 theme molecule 586:593 arg1 shape 613:617 the molecule a "mushroom-like" shape 582:617 the molecule a "mushroom-like" shape 582:617 This structure shows a domain consisting of two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape. 12657638 0 32 theme functional 46:55 arg1 roles 57:61 functional roles 46:61 functional roles of membrane association 46:85 Structure of human estrone sulfatase suggests functional roles of membrane association. 12657638 1 33 theme endoplasmic 267:277 arg1 ER 290:291 ER 290:291 ER 290:291 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 33 theme endoplasmic 267:277 arg1 reticulum 279:287 the endoplasmic reticulum 263:287 the endoplasmic reticulum (ER) 263:292 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 4 34 theme functional 746:755 arg1 domain 757:762 the functional domain 742:762 the functional domain on the membrane surface facing the ER lumen 742:806 These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. 12657638 5 35 theme transmembrane 829:841 arg1 domain 843:848 the transmembrane domain 825:848 the transmembrane domain 825:848 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 1 36 theme reticulum 279:287 arg1 membrane 251:258 the membrane 247:258 the membrane of the endoplasmic reticulum (ER) 247:292 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 5 37 theme gill 927:930 arg1 " 931:931 the "gill" 922:931 the "gill" of the "mushroom 922:948 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 6 38 theme simple 1051:1056 arg1 prototype 1082:1090 a prototype 1080:1090 a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes 1080:1180 This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes. 12657638 6 38 theme simple 1051:1056 arg1 architecture 1058:1069 This simple architecture 1046:1069 This simple architecture 1046:1069 This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes. 12657638 1 39 theme amino 115:119 arg1 acids 121:125 562 amino acids 111:125 562 amino acids 111:125 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 3 40 theme mushroom-like 598:610 arg1 shape 613:617 the molecule a "mushroom-like" shape 582:617 the molecule a "mushroom-like" shape 582:617 This structure shows a domain consisting of two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape. 12657638 5 41 theme buried 895:900 arg1 deep 902:905 buried deep 895:905 buried deep in a cavity of the "gill" of the "mushroom 895:948 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 5 41 theme buried 895:900 arg1 opening 867:873 the opening 863:873 the opening to the active site 863:892 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 1 42 theme estrogens 195:203 arg1 levels 185:190 high levels 180:190 high levels of estrogens 180:203 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 0 43 theme membrane 66:73 arg1 association 75:85 membrane association 66:85 membrane association 66:85 Structure of human estrone sulfatase suggests functional roles of membrane association. 12657638 3 44 theme " 611:611 arg1 shape 613:617 the molecule a "mushroom-like" shape 582:617 the molecule a "mushroom-like" shape 582:617 This structure shows a domain consisting of two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape. 12657638 2 45 theme ES 312:313 arg1 structure 299:307 The structure 295:307 The structure of ES, purified from the microsomal fraction of human placentas, 295:372 The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. 12657638 5 46 theme membrane 967:974 arg1 surface 976:982 the membrane surface 963:982 the membrane surface 963:982 The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. 12657638 0 47 theme human 13:17 arg1 sulfatase 27:35 human estrone sulfatase 13:35 human estrone sulfatase 13:35 Structure of human estrone sulfatase suggests functional roles of membrane association. 12657638 1 48 dep ES 107:108 arg1 acids 121:125 562 amino acids 111:125 562 amino acids 111:125 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 12657638 1 49 theme breast 208:213 arg1 cells 221:225 breast tumor cells 208:225 breast tumor cells 208:225 Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). 15696168 5 0 theme new 802:804 arg1 constraints 806:816 new constraints 802:816 new constraints 802:816 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 6 1 theme CD28-CTLA-4-CD80-CD86 936:956 arg1 system 968:973 the CD28-CTLA-4-CD80-CD86 signaling system 932:973 the CD28-CTLA-4-CD80-CD86 signaling system 932:973 This work completes the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system. 15696168 1 2 theme cell 136:139 arg1 receptors 149:157 nonclonotypic cell surface receptors 122:157 nonclonotypic cell surface receptors 122:157 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 5 3 theme nonmitogenic 772:783 arg1 antibodies 785:794 mitogenic and nonmitogenic antibodies 758:794 mitogenic and nonmitogenic antibodies 758:794 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 4 4 theme proteins 510:517 arg1 relationships 480:492 the evolutionary relationships 463:492 the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules 463:559 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 1 5 theme surface 141:147 arg1 receptors 149:157 nonclonotypic cell surface receptors 122:157 nonclonotypic cell surface receptors 122:157 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 4 6 theme CD28-related 497:508 arg1 proteins 510:517 CD28-related proteins 497:517 CD28-related proteins 497:517 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 1 7 theme Naive 49:53 arg1 activation 62:71 Naive T cell activation 49:71 Naive T cell activation 49:71 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 4 8 theme ligand-binding 590:603 arg1 properties 624:633 the distinct ligand-binding and stoichiometric properties 577:633 the distinct ligand-binding and stoichiometric properties of CD28 577:641 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 5 9 theme mitogenic 758:766 arg1 antibodies 785:794 mitogenic and nonmitogenic antibodies 758:794 mitogenic and nonmitogenic antibodies 758:794 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 3 10 with complex 376:382 arg1 fragment 397:404 the Fab fragment 389:404 the Fab fragment of a mitogenic antibody 389:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 1 11 theme T 55:55 arg1 activation 62:71 Naive T cell activation 49:71 Naive T cell activation 49:71 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 4 12 theme distinct 581:588 arg1 properties 624:633 the distinct ligand-binding and stoichiometric properties 577:633 the distinct ligand-binding and stoichiometric properties of CD28 577:641 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 3 13 theme crystal 329:335 arg1 structure 337:345 the crystal structure 325:345 the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody 325:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 4 14 theme related 651:657 arg1 CTLA-4 681:686 the related, inhibitory homodimer CTLA-4 647:686 CTLA-4 681:686 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 1 15 theme cell 57:60 arg1 activation 62:71 Naive T cell activation 49:71 Naive T cell activation 49:71 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 4 16 theme inhibitory 660:669 arg1 CTLA-4 681:686 the related, inhibitory homodimer CTLA-4 647:686 CTLA-4 681:686 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 5 17 theme antibody-induced 831:846 arg1 triggering 857:866 antibody-induced receptor triggering 831:866 antibody-induced receptor triggering 831:866 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 4 18 theme evolutionary 467:478 arg1 relationships 480:492 the evolutionary relationships 463:492 the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules 463:559 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 3 19 theme CD28 368:371 arg1 form 360:363 a soluble form 350:363 a soluble form of CD28 350:371 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 5 20 with comparisons 720:730 arg1 antibodies 785:794 mitogenic and nonmitogenic antibodies 758:794 mitogenic and nonmitogenic antibodies 758:794 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 5 21 theme receptor 848:855 arg1 triggering 857:866 antibody-induced receptor triggering 831:866 antibody-induced receptor triggering 831:866 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 5 22 from place 796:800 arg1 models 821:826 models 821:826 models of antibody-induced receptor triggering 821:866 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 6 23 theme system 968:973 arg1 characterization 912:927 the initial structural characterization 889:927 the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system 889:973 This work completes the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system. 15696168 3 24 theme Fab 393:395 arg1 fragment 397:404 the Fab fragment 389:404 the Fab fragment of a mitogenic antibody 389:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 0 25 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a soluble CD28-Fab complex 0:46 Crystal structure of a soluble CD28-Fab complex. 15696168 4 26 theme molecules 551:559 arg1 relationships 480:492 the evolutionary relationships 463:492 the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules 463:559 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 2 27 theme important 169:177 arg1 homodimer 219:227 the monovalent homodimer CD28 204:232 the monovalent homodimer CD28 204:232 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 2 27 theme important 169:177 arg1 protein 193:199 The most important costimulatory protein 160:199 The most important costimulatory protein 160:199 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 5 28 theme triggering 857:866 arg1 models 821:826 models 821:826 models of antibody-induced receptor triggering 821:866 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 4 29 theme homodimer 671:679 arg1 CTLA-4 681:686 the related, inhibitory homodimer CTLA-4 647:686 CTLA-4 681:686 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 3 30 theme soluble 352:358 arg1 form 360:363 a soluble form 350:363 a soluble form of CD28 350:371 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 4 31 theme adhesion 542:549 arg1 molecules 551:559 adhesion molecules 542:559 adhesion molecules 542:559 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 2 32 theme antigen-presenting 283:300 arg1 cells 302:306 antigen-presenting cells 283:306 antigen-presenting cells 283:306 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 3 33 theme form 360:363 arg1 structure 337:345 the crystal structure 325:345 the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody 325:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 5 34 theme complexes 735:743 arg1 comparisons 720:730 Cryo-electron microscopy-based comparisons 689:730 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies 689:794 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 4 35 theme CD28 638:641 arg1 properties 624:633 the distinct ligand-binding and stoichiometric properties 577:633 the distinct ligand-binding and stoichiometric properties of CD28 577:641 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 4 35 theme CD28 638:641 arg1 CTLA-4 681:686 the related, inhibitory homodimer CTLA-4 647:686 CTLA-4 681:686 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 0 36 theme CD28-Fab 31:38 arg1 complex 40:46 a soluble CD28-Fab complex 21:46 a soluble CD28-Fab complex 21:46 Crystal structure of a soluble CD28-Fab complex. 15696168 3 37 from structure 337:345 arg1 complex 376:382 complex 376:382 complex with the Fab fragment of a mitogenic antibody 376:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 3 38 theme mitogenic 411:419 arg1 antibody 421:428 a mitogenic antibody 409:428 a mitogenic antibody 409:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 0 39 theme soluble 23:29 arg1 complex 40:46 a soluble CD28-Fab complex 21:46 a soluble CD28-Fab complex 21:46 Crystal structure of a soluble CD28-Fab complex. 15696168 4 40 theme antigen 520:526 arg1 receptors 528:536 antigen receptors 520:536 antigen receptors 520:536 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 5 41 theme CD28 748:751 arg1 complexes 735:743 complexes 735:743 complexes of CD28 735:751 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 4 42 theme Structural 431:440 arg1 comparisons 442:452 Structural comparisons 431:452 Structural comparisons 431:452 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 2 43 dep homodimer 219:227 arg1 CD28 229:232 CD28 229:232 the monovalent homodimer CD28 204:232 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 6 44 theme structural 901:910 arg1 characterization 912:927 the initial structural characterization 889:927 the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system 889:973 This work completes the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system. 15696168 2 45 theme monovalent 208:217 arg1 homodimer 219:227 the monovalent homodimer CD28 204:232 the monovalent homodimer CD28 204:232 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 2 45 theme monovalent 208:217 arg1 protein 193:199 The most important costimulatory protein 160:199 The most important costimulatory protein 160:199 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 6 46 theme signaling 958:966 arg1 system 968:973 the CD28-CTLA-4-CD80-CD86 signaling system 932:973 the CD28-CTLA-4-CD80-CD86 signaling system 932:973 This work completes the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system. 15696168 1 47 theme T 99:99 arg1 receptor 106:113 the T cell receptor 95:113 the T cell receptor 95:113 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 0 48 theme complex 40:46 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a soluble CD28-Fab complex 0:46 Crystal structure of a soluble CD28-Fab complex. 15696168 5 49 theme Cryo-electron 689:701 arg1 comparisons 720:730 Cryo-electron microscopy-based comparisons 689:730 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies 689:794 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 6 50 theme initial 893:899 arg1 characterization 912:927 the initial structural characterization 889:927 the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system 889:973 This work completes the initial structural characterization of the CD28-CTLA-4-CD80-CD86 signaling system. 15696168 1 51 theme cell 101:104 arg1 receptor 106:113 the T cell receptor 95:113 the T cell receptor 95:113 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 15696168 4 52 theme receptors 528:536 arg1 relationships 480:492 the evolutionary relationships 463:492 the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules 463:559 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 5 53 theme microscopy-based 703:718 arg1 comparisons 720:730 Cryo-electron microscopy-based comparisons 689:730 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies 689:794 Cryo-electron microscopy-based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. 15696168 4 54 theme stoichiometric 609:622 arg1 properties 624:633 the distinct ligand-binding and stoichiometric properties 577:633 the distinct ligand-binding and stoichiometric properties of CD28 577:641 Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. 15696168 2 55 theme costimulatory 179:191 arg1 homodimer 219:227 the monovalent homodimer CD28 204:232 the monovalent homodimer CD28 204:232 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 2 55 theme costimulatory 179:191 arg1 protein 193:199 The most important costimulatory protein 160:199 The most important costimulatory protein 160:199 The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. 15696168 3 56 theme antibody 421:428 arg1 fragment 397:404 the Fab fragment 389:404 the Fab fragment of a mitogenic antibody 389:428 Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. 15696168 1 57 theme nonclonotypic 122:134 arg1 receptors 149:157 nonclonotypic cell surface receptors 122:157 nonclonotypic cell surface receptors 122:157 Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. 21692750 6 0 theme dileucine 1236:1244 arg1 motif 1246:1250 a N-terminal dileucine motif 1223:1250 a N-terminal dileucine motif 1223:1250 Lysosomal targeting of DIRC2 was demonstrated to be mediated by a N-terminal dileucine motif. 21692750 8 1 theme mixture 1518:1524 arg1 application 1483:1493 the application 1479:1493 the application of a complex metabolic mixture 1479:1524 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 8 2 theme metabolic 1508:1516 arg1 mixture 1518:1524 a complex metabolic mixture 1498:1524 a complex metabolic mixture 1498:1524 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 0 3 theme novel 42:46 arg1 transporter 48:58 a novel transporter 40:58 a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) 40:169 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 0 3 theme novel 42:46 arg1 gene 226:229 a breakpoint-spanning gene 204:229 a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer 204:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 4 4 theme murine 909:914 arg1 fibroblasts 926:936 murine embryonic fibroblasts 909:936 murine embryonic fibroblasts 909:936 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 0 5 theme renal 152:156 arg1 carcinoma 158:166 renal carcinoma 2 152:168 renal carcinoma 2 152:168 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 0 6 dep membrane 77:84 arg1 processed 106:114 processed 106:114 is proteolytically processed by cathepsin L. DIRC2 87:136 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 3 7 theme C-terminal 711:720 arg1 fragment 722:729 a non-glycosylated C-terminal fragment 692:729 a non-glycosylated C-terminal fragment 692:729 DIRC2 is proteolytically processed into a N-glycosylated N-terminal and a non-glycosylated C-terminal fragment respectively. 21692750 5 8 theme cleavage 943:950 arg1 site 952:955 The cleavage site 939:955 The cleavage site within DIRC2 939:968 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 5 8 theme cleavage 943:950 arg1 located 1068:1074 located 1068:1074 located 1068:1074 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 3 9 theme non-glycosylated 694:709 arg1 fragment 722:729 a non-glycosylated C-terminal fragment 692:729 a non-glycosylated C-terminal fragment 692:729 DIRC2 is proteolytically processed into a N-glycosylated N-terminal and a non-glycosylated C-terminal fragment respectively. 21692750 2 10 theme overexpressed 545:557 arg1 residence 532:540 lysosomal residence 522:540 lysosomal residence of overexpressed as well as endogenous DIRC2 522:585 In the present study, lysosomal residence of overexpressed as well as endogenous DIRC2 was shown by several approaches. 21692750 2 11 theme several 600:606 arg1 approaches 608:617 several approaches 600:617 several approaches 600:617 In the present study, lysosomal residence of overexpressed as well as endogenous DIRC2 was shown by several approaches. 21692750 0 12 theme renal 308:312 arg1 cancer 314:319 renal cancer 308:319 renal cancer 308:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 0 13 theme membrane 77:84 arg1 transporter 48:58 a novel transporter 40:58 a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) 40:169 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 0 13 theme membrane 77:84 arg1 gene 226:229 a breakpoint-spanning gene 204:229 a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer 204:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 1 14 theme DIRC2 326:330 arg1 protein 332:338 The DIRC2 protein 322:338 The DIRC2 protein 322:338 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 4 15 theme L 851:851 arg1 activity 829:836 the activity 825:836 the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts 825:936 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 4 16 theme lysosomal 776:784 arg1 compartments 786:797 lysosomal compartments 776:797 lysosomal compartments 776:797 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 7 17 theme plasma 1310:1315 arg1 membrane 1317:1324 the plasma membrane 1306:1324 the plasma membrane 1306:1324 By disrupting this motif, DIRC2 can be redirected to the plasma membrane. 21692750 0 18 theme lysosomal 67:75 arg1 membrane 77:84 the lysosomal membrane 63:84 the lysosomal membrane 63:84 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 5 19 theme fifth 1097:1101 arg1 loop 1118:1121 the tentative fifth intralysosomal loop 1083:1121 the tentative fifth intralysosomal loop 1083:1121 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 4 20 theme cathepsin 841:849 arg1 L 851:851 cathepsin L 841:851 cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts 841:936 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 0 21 theme breakpoint-spanning 206:224 arg1 transporter 48:58 a novel transporter 40:58 a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) 40:169 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 0 21 theme breakpoint-spanning 206:224 arg1 gene 226:229 a breakpoint-spanning gene 204:229 a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer 204:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 6 22 theme Lysosomal 1159:1167 arg1 targeting 1169:1177 Lysosomal targeting 1159:1177 Lysosomal targeting of DIRC2 1159:1186 Lysosomal targeting of DIRC2 was demonstrated to be mediated by a N-terminal dileucine motif. 21692750 5 23 theme tentative 1087:1095 arg1 loop 1118:1121 the tentative fifth intralysosomal loop 1083:1121 the tentative fifth intralysosomal loop 1083:1121 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 9 24 theme proteolytic 1787:1797 arg1 processing 1799:1808 limited proteolytic processing 1779:1808 limited proteolytic processing 1779:1808 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 5 25 theme intralysosomal 1103:1116 arg1 loop 1118:1121 the tentative fifth intralysosomal loop 1083:1121 the tentative fifth intralysosomal loop 1083:1121 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 2 26 theme DIRC2 581:585 arg1 residence 532:540 lysosomal residence 522:540 lysosomal residence of overexpressed as well as endogenous DIRC2 522:585 In the present study, lysosomal residence of overexpressed as well as endogenous DIRC2 was shown by several approaches. 21692750 8 27 theme whole-cell 1341:1350 arg1 assay 1373:1377 a whole-cell electrophysiological assay 1339:1377 a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes 1339:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 0 28 theme cancer 314:319 arg1 development 293:303 the development 289:303 the development of renal cancer 289:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 8 29 theme outward 1536:1542 arg1 current 1544:1550 an outward current 1533:1550 an outward current associated with the surface expression of full-length DIRC2 1533:1610 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 2 30 theme endogenous 570:579 arg1 DIRC2 581:585 endogenous DIRC2 570:585 overexpressed as well as endogenous DIRC2 545:585 In the present study, lysosomal residence of overexpressed as well as endogenous DIRC2 was shown by several approaches. 21692750 1 31 theme organellar 427:436 arg1 proteomics 438:447 organellar proteomics 427:447 organellar proteomics as a tentative constituent of lysosomal membranes 427:497 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 5 32 theme epitope 1036:1042 arg1 tags 1044:1047 internal epitope tags 1027:1047 internal epitope tags 1027:1047 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 5 33 located located 1068:1074 arg2 located 1068:1074 located 1068:1074 located 1068:1074 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 5 33 located located 1068:1074 arg1 loop 1118:1121 the tentative fifth intralysosomal loop 1083:1121 the tentative fifth intralysosomal loop 1083:1121 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 5 33 located located 1068:1074 arg2 site 952:955 The cleavage site 939:955 The cleavage site within DIRC2 939:968 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 9 34 theme limited 1779:1785 arg1 processing 1799:1808 limited proteolytic processing 1779:1808 limited proteolytic processing 1779:1808 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 3 35 gly non-glycosylated 694:709 arg1 fragment 722:729 a non-glycosylated C-terminal fragment 692:729 a non-glycosylated C-terminal fragment 692:729 DIRC2 is proteolytically processed into a N-glycosylated N-terminal and a non-glycosylated C-terminal fragment respectively. 21692750 4 36 theme embryonic 916:924 arg1 fibroblasts 926:936 murine embryonic fibroblasts 909:936 murine embryonic fibroblasts 909:936 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 5 37 theme internal 1027:1034 arg1 tags 1044:1047 internal epitope tags 1027:1047 internal epitope tags 1027:1047 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 4 38 theme Proteolytic 745:755 arg1 cleavage 757:764 Proteolytic cleavage 745:764 Proteolytic cleavage 745:764 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 0 39 theme renal 13:17 arg1 carcinoma 19:27 renal carcinoma 2 13:29 renal carcinoma 2 (DIRC2) 13:37 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 0 39 theme renal 13:17 arg1 DIRC2 32:36 DIRC2 32:36 DIRC2 32:36 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 8 40 theme complex 1500:1506 arg1 mixture 1518:1524 a complex metabolic mixture 1498:1524 a complex metabolic mixture 1498:1524 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 2 41 theme present 507:513 arg1 study 515:519 the present study 503:519 the present study 503:519 In the present study, lysosomal residence of overexpressed as well as endogenous DIRC2 was shown by several approaches. 21692750 6 42 theme DIRC2 1182:1186 arg1 targeting 1169:1177 Lysosomal targeting 1159:1177 Lysosomal targeting of DIRC2 1159:1186 Lysosomal targeting of DIRC2 was demonstrated to be mediated by a N-terminal dileucine motif. 21692750 8 43 theme electrophysiological 1352:1371 arg1 assay 1373:1377 a whole-cell electrophysiological assay 1339:1377 a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes 1339:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 3 44 dep N-terminal 677:686 arg1 a 660:660 a 660:660 a 660:660 DIRC2 is proteolytically processed into a N-glycosylated N-terminal and a non-glycosylated C-terminal fragment respectively. 21692750 9 45 theme lysosomal 1696:1704 arg1 DIRC2 1671:1675 DIRC2 1671:1675 DIRC2 1671:1675 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 9 45 theme lysosomal 1696:1704 arg1 transporter 1717:1727 an electrogenic lysosomal metabolite transporter 1680:1727 an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing 1680:1808 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 6 46 theme N-terminal 1225:1234 arg1 motif 1246:1250 a N-terminal dileucine motif 1223:1250 a N-terminal dileucine motif 1223:1250 Lysosomal targeting of DIRC2 was demonstrated to be mediated by a N-terminal dileucine motif. 21692750 5 47 theme typical 1137:1143 arg1 topology 1149:1156 the typical MFS topology 1133:1156 the typical MFS topology 1133:1156 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 9 48 theme metabolite 1706:1715 arg1 DIRC2 1671:1675 DIRC2 1671:1675 DIRC2 1671:1675 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 9 48 theme metabolite 1706:1715 arg1 transporter 1717:1727 an electrogenic lysosomal metabolite transporter 1680:1727 an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing 1680:1808 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 0 49 theme chromosomal 236:246 arg1 translocation 248:260 a chromosomal translocation 234:260 a chromosomal translocation putatively associated with the development of renal cancer 234:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 8 50 theme Xenopus 1462:1468 arg1 oocytes 1470:1476 Xenopus oocytes 1462:1476 Xenopus oocytes 1462:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 5 51 theme MFS 1145:1147 arg1 topology 1149:1156 the typical MFS topology 1133:1156 the typical MFS topology 1133:1156 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 0 52 theme cathepsin 119:127 arg1 DIRC2 132:136 cathepsin L. DIRC2 119:136 cathepsin L. DIRC2 119:136 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 8 53 theme plasma 1443:1448 arg1 membrane 1450:1457 the plasma membrane 1439:1457 the plasma membrane of Xenopus oocytes 1439:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 1 54 theme major 360:364 arg1 MFS 355:357 the MFS 351:357 the MFS (major facilitator superfamily) 351:389 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 1 54 theme major 360:364 arg1 superfamily 378:388 major facilitator superfamily 360:388 major facilitator superfamily 360:388 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 4 55 from process 898:904 arg1 fibroblasts 926:936 murine embryonic fibroblasts 909:936 murine embryonic fibroblasts 909:936 Proteolytic cleavage occurs in lysosomal compartments and critically depends on the activity of cathepsin L which was found to be indispensable for this process in murine embryonic fibroblasts. 21692750 1 56 theme tentative 454:462 arg1 constituent 464:474 a tentative constituent 452:474 a tentative constituent of lysosomal membranes 452:497 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 8 57 theme surface 1572:1578 arg1 expression 1580:1589 the surface expression 1568:1589 the surface expression of full-length DIRC2 1568:1610 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 2 58 theme lysosomal 522:530 arg1 residence 532:540 lysosomal residence 522:540 lysosomal residence of overexpressed as well as endogenous DIRC2 522:585 In the present study, lysosomal residence of overexpressed as well as endogenous DIRC2 was shown by several approaches. 21692750 1 59 theme facilitator 366:376 arg1 MFS 355:357 the MFS 351:357 the MFS (major facilitator superfamily) 351:389 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 1 59 theme facilitator 366:376 arg1 superfamily 378:388 major facilitator superfamily 360:388 major facilitator superfamily 360:388 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 5 60 theme amino 989:993 arg1 residues 1000:1007 amino acid residues 214 and 261 989:1019 residues 1000:1007 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 8 61 theme full-length 1594:1604 arg1 DIRC2 1606:1610 full-length DIRC2 1594:1610 full-length DIRC2 1594:1610 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 0 62 theme L. 129:130 arg1 DIRC2 132:136 cathepsin L. DIRC2 119:136 cathepsin L. DIRC2 119:136 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 5 63 theme acid 995:998 arg1 residues 1000:1007 amino acid residues 214 and 261 989:1019 residues 1000:1007 The cleavage site within DIRC2 was mapped between amino acid residues 214 and 261 using internal epitope tags, and is presumably located within the tentative fifth intralysosomal loop, assuming the typical MFS topology. 21692750 8 64 from membrane 1450:1457 arg1 expression 1401:1410 heterologous expression 1388:1410 heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes 1388:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 8 65 theme oocytes 1470:1476 arg1 membrane 1450:1457 the plasma membrane 1439:1457 the plasma membrane of Xenopus oocytes 1439:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 8 66 theme mutant 1429:1434 arg1 expression 1401:1410 heterologous expression 1388:1410 heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes 1388:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 1 67 theme lysosomal 479:487 arg1 membranes 489:497 lysosomal membranes 479:497 lysosomal membranes 479:497 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 8 68 theme heterologous 1388:1399 arg1 expression 1401:1410 heterologous expression 1388:1410 heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes 1388:1476 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 8 69 theme targeting 1419:1427 arg1 mutant 1429:1434 the targeting mutant 1415:1434 the targeting mutant 1415:1434 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 21692750 1 70 theme membranes 489:497 arg1 constituent 464:474 a tentative constituent 452:474 a tentative constituent of lysosomal membranes 452:497 The DIRC2 protein belongs to the MFS (major facilitator superfamily) and has been previously detected by organellar proteomics as a tentative constituent of lysosomal membranes. 21692750 0 71 from gene 226:229 arg1 translocation 248:260 a chromosomal translocation 234:260 a chromosomal translocation putatively associated with the development of renal cancer 234:319 Disrupted in renal carcinoma 2 (DIRC2), a novel transporter of the lysosomal membrane, is proteolytically processed by cathepsin L. DIRC2 (Disrupted in renal carcinoma 2) has been initially identified as a breakpoint-spanning gene in a chromosomal translocation putatively associated with the development of renal cancer. 21692750 9 72 theme electrogenic 1683:1694 arg1 DIRC2 1671:1675 DIRC2 1671:1675 DIRC2 1671:1675 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 9 72 theme electrogenic 1683:1694 arg1 transporter 1717:1727 an electrogenic lysosomal metabolite transporter 1680:1727 an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing 1680:1808 Taken together, these data strongly support the idea that DIRC2 is an electrogenic lysosomal metabolite transporter which is subjected to and presumably modulated by limited proteolytic processing. 21692750 8 73 theme DIRC2 1606:1610 arg1 expression 1580:1589 the surface expression 1568:1589 the surface expression of full-length DIRC2 1568:1610 Finally, in a whole-cell electrophysiological assay based on heterologous expression of the targeting mutant at the plasma membrane of Xenopus oocytes, the application of a complex metabolic mixture evokes an outward current associated with the surface expression of full-length DIRC2. 19153605 9 0 theme LPS 1102:1104 arg1 binding 1106:1112 LPS binding 1102:1112 LPS binding 1102:1112 We further showed that mindin recognizes lipopolysaccharide (LPS) through its TSR domain, and obtained evidence that C-mannosylation of the TSR influences LPS binding. 19153605 6 1 theme C2 730:731 arg1 domains 733:739 membrane-targeting C2 domains 711:739 membrane-targeting C2 domains 711:739 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 6 1 theme C2 730:731 arg1 ion 768:770 a bound calcium ion 752:770 a bound calcium ion 752:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 1 2 theme extracellular 123:135 arg1 protein 144:150 an extracellular matrix protein 120:150 an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells 120:236 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 1 2 theme extracellular 123:135 arg1 Mindin 98:103 Mindin 98:103 Mindin (spondin-2) 98:115 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 1 3 theme dendritic 222:230 arg1 cells 232:236 dendritic cells 222:236 dendritic cells 222:236 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 0 4 theme ligand 57:62 arg1 Structure 0:8 Structure 0:8 Structure of the F-spondin domain of mindin 0:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 0 4 theme ligand 57:62 arg1 molecule 88:95 an integrin ligand and pattern recognition molecule 45:95 an integrin ligand and pattern recognition molecule 45:95 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 4 5 theme C-terminal 505:514 arg1 TSR 546:548 TSR 546:548 TSR 546:548 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 4 5 theme C-terminal 505:514 arg1 repeat 538:543 C-terminal thrombospondin type 1 repeat 505:543 C-terminal thrombospondin type 1 repeat (TSR) 505:549 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 1 6 theme matrix 137:142 arg1 protein 144:150 an extracellular matrix protein 120:150 an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells 120:236 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 1 6 theme matrix 137:142 arg1 Mindin 98:103 Mindin 98:103 Mindin (spondin-2) 98:115 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 0 7 theme recognition 76:86 arg1 Structure 0:8 Structure 0:8 Structure of the F-spondin domain of mindin 0:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 0 7 theme recognition 76:86 arg1 molecule 88:95 an integrin ligand and pattern recognition molecule 45:95 an integrin ligand and pattern recognition molecule 45:95 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 3 8 with interactions 381:392 arg1 integrins 399:407 integrins 399:407 integrins 399:407 These dual functions are mediated by interactions with integrins and microbial pathogens, respectively. 19153605 3 8 with interactions 381:392 arg1 pathogens 423:431 microbial pathogens 413:431 microbial pathogens 413:431 These dual functions are mediated by interactions with integrins and microbial pathogens, respectively. 19153605 10 9 theme responses 1235:1243 arg1 activation 1189:1198 activation 1189:1198 activation of both adaptive and innate immune responses 1189:1243 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 3 10 theme dual 350:353 arg1 functions 355:363 These dual functions 344:363 These dual functions 344:363 These dual functions are mediated by interactions with integrins and microbial pathogens, respectively. 19153605 4 11 theme F-spondin 479:487 arg1 domain 494:499 an N-terminal F-spondin (FS) domain 465:499 an N-terminal F-spondin (FS) domain 465:499 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 5 12 theme FS 587:588 arg1 domain 590:595 the FS domain 583:595 the FS domain 583:595 We determined the structure of the FS domain at 1.8-A resolution. 19153605 7 13 theme integrin 817:824 arg1 binding 826:832 integrin binding 817:832 integrin binding 817:832 We demonstrated that the FS domain mediates integrin binding and identified the binding site by mutagenesis. 19153605 6 14 theme membrane-targeting 711:728 arg1 domains 733:739 membrane-targeting C2 domains 711:739 membrane-targeting C2 domains 711:739 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 6 14 theme membrane-targeting 711:728 arg1 ion 768:770 a bound calcium ion 752:770 a bound calcium ion 752:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 9 15 theme TSR 1087:1089 arg1 C-mannosylation 1064:1078 C-mannosylation 1064:1078 C-mannosylation of the TSR 1064:1089 We further showed that mindin recognizes lipopolysaccharide (LPS) through its TSR domain, and obtained evidence that C-mannosylation of the TSR influences LPS binding. 19153605 4 16 theme N-terminal 468:477 arg1 domain 494:499 an N-terminal F-spondin (FS) domain 465:499 an N-terminal F-spondin (FS) domain 465:499 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 5 17 theme domain 590:595 arg1 structure 570:578 the structure 566:578 the structure of the FS domain at 1.8-A resolution 566:615 We determined the structure of the FS domain at 1.8-A resolution. 19153605 1 18 theme unknown 155:161 arg1 structure 163:171 unknown structure 155:171 unknown structure 155:171 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 4 19 theme FS 490:491 arg1 domain 494:499 an N-terminal F-spondin (FS) domain 465:499 an N-terminal F-spondin (FS) domain 465:499 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 5 20 theme 1.8-A 600:604 arg1 resolution 606:615 1.8-A resolution 600:615 1.8-A resolution 600:615 We determined the structure of the FS domain at 1.8-A resolution. 19153605 8 21 theme mindin 886:891 arg1 domain 896:901 The mindin FS domain 882:901 The mindin FS domain 882:901 The mindin FS domain therefore represents a new integrin ligand. 19153605 7 22 theme binding 853:859 arg1 site 861:864 the binding site 849:864 the binding site 849:864 We demonstrated that the FS domain mediates integrin binding and identified the binding site by mutagenesis. 19153605 3 23 theme microbial 413:421 arg1 pathogens 423:431 microbial pathogens 413:431 microbial pathogens 413:431 These dual functions are mediated by interactions with integrins and microbial pathogens, respectively. 19153605 5 24 from resolution 606:615 arg1 structure 570:578 the structure 566:578 the structure of the FS domain at 1.8-A resolution 566:615 We determined the structure of the FS domain at 1.8-A resolution. 19153605 2 25 theme recognition 283:293 arg1 mindin 253:258 mindin 253:258 mindin 253:258 Additionally, mindin functions as a pattern recognition molecule for initiating innate immune responses. 19153605 2 25 theme recognition 283:293 arg1 molecule 295:302 a pattern recognition molecule 273:302 a pattern recognition molecule for initiating innate immune responses 273:341 Additionally, mindin functions as a pattern recognition molecule for initiating innate immune responses. 19153605 1 26 theme structure 163:171 arg1 protein 144:150 an extracellular matrix protein 120:150 an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells 120:236 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 1 26 theme structure 163:171 arg1 Mindin 98:103 Mindin 98:103 Mindin (spondin-2) 98:115 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 0 27 theme F-spondin 17:25 arg1 mindin 37:42 mindin 37:42 mindin 37:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 0 27 theme F-spondin 17:25 arg1 domain 27:32 the F-spondin domain 13:32 the F-spondin domain of mindin 13:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 10 28 theme dual 1129:1132 arg1 interactions 1134:1145 these dual interactions 1123:1145 these dual interactions 1123:1145 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 10 29 theme FS 1152:1153 arg1 mindin 1174:1179 mindin 1174:1179 mindin 1174:1179 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 10 29 theme FS 1152:1153 arg1 domains 1163:1169 the FS and TSR domains 1148:1169 the FS and TSR domains of mindin 1148:1179 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 2 30 theme pattern 275:281 arg1 mindin 253:258 mindin 253:258 mindin 253:258 Additionally, mindin functions as a pattern recognition molecule for initiating innate immune responses. 19153605 2 30 theme pattern 275:281 arg1 molecule 295:302 a pattern recognition molecule 273:302 a pattern recognition molecule for initiating innate immune responses 273:341 Additionally, mindin functions as a pattern recognition molecule for initiating innate immune responses. 19153605 6 31 theme beta-sandwich 672:684 arg1 motif 686:690 an eight-stranded antiparallel beta-sandwich motif 641:690 an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion 641:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 10 32 theme innate 1221:1226 arg1 responses 1235:1243 both adaptive and innate immune responses 1203:1243 both adaptive and innate immune responses 1203:1243 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 9 33 gly C-mannosylation 1064:1078 arg1 TSR 1087:1089 the TSR 1083:1089 the TSR 1083:1089 We further showed that mindin recognizes lipopolysaccharide (LPS) through its TSR domain, and obtained evidence that C-mannosylation of the TSR influences LPS binding. 19153605 6 34 theme antiparallel 659:670 arg1 motif 686:690 an eight-stranded antiparallel beta-sandwich motif 641:690 an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion 641:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 7 35 theme FS 798:799 arg1 domain 801:806 the FS domain 794:806 the FS domain 794:806 We demonstrated that the FS domain mediates integrin binding and identified the binding site by mutagenesis. 19153605 6 36 theme eight-stranded 644:657 arg1 motif 686:690 an eight-stranded antiparallel beta-sandwich motif 641:690 an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion 641:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 0 37 theme domain 27:32 arg1 Structure 0:8 Structure 0:8 Structure of the F-spondin domain of mindin 0:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 0 37 theme domain 27:32 arg1 molecule 88:95 an integrin ligand and pattern recognition molecule 45:95 an integrin ligand and pattern recognition molecule 45:95 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 4 38 theme type 531:534 arg1 TSR 546:548 TSR 546:548 TSR 546:548 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 4 38 theme type 531:534 arg1 repeat 538:543 C-terminal thrombospondin type 1 repeat 505:543 C-terminal thrombospondin type 1 repeat (TSR) 505:549 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 8 39 theme new 926:928 arg1 ligand 939:944 a new integrin ligand 924:944 a new integrin ligand 924:944 The mindin FS domain therefore represents a new integrin ligand. 19153605 4 40 theme thrombospondin 516:529 arg1 TSR 546:548 TSR 546:548 TSR 546:548 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 4 40 theme thrombospondin 516:529 arg1 repeat 538:543 C-terminal thrombospondin type 1 repeat 505:543 C-terminal thrombospondin type 1 repeat (TSR) 505:549 Mindin comprises an N-terminal F-spondin (FS) domain and C-terminal thrombospondin type 1 repeat (TSR). 19153605 6 41 theme calcium 760:766 arg1 ion 768:770 a bound calcium ion 752:770 a bound calcium ion 752:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 2 42 theme immune 326:331 arg1 responses 333:341 innate immune responses 319:341 innate immune responses 319:341 Additionally, mindin functions as a pattern recognition molecule for initiating innate immune responses. 19153605 0 43 theme mindin 37:42 arg1 mindin 37:42 mindin 37:42 mindin 37:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 0 43 theme mindin 37:42 arg1 domain 27:32 the F-spondin domain 13:32 the F-spondin domain of mindin 13:42 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 6 44 theme bound 754:758 arg1 ion 768:770 a bound calcium ion 752:770 a bound calcium ion 752:770 The structure revealed an eight-stranded antiparallel beta-sandwich motif resembling that of membrane-targeting C2 domains, including a bound calcium ion. 19153605 2 45 theme innate 319:324 arg1 responses 333:341 innate immune responses 319:341 innate immune responses 319:341 Additionally, mindin functions as a pattern recognition molecule for initiating innate immune responses. 19153605 10 46 theme immune 1228:1233 arg1 responses 1235:1243 both adaptive and innate immune responses 1203:1243 both adaptive and innate immune responses 1203:1243 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 8 47 theme integrin 930:937 arg1 ligand 939:944 a new integrin ligand 924:944 a new integrin ligand 924:944 The mindin FS domain therefore represents a new integrin ligand. 19153605 9 48 theme TSR 1025:1027 arg1 domain 1029:1034 its TSR domain 1021:1034 its TSR domain 1021:1034 We further showed that mindin recognizes lipopolysaccharide (LPS) through its TSR domain, and obtained evidence that C-mannosylation of the TSR influences LPS binding. 19153605 1 49 theme efficient 194:202 arg1 priming 211:217 efficient T-cell priming 194:217 efficient T-cell priming by dendritic cells 194:236 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 8 50 theme FS 893:894 arg1 domain 896:901 The mindin FS domain 882:901 The mindin FS domain 882:901 The mindin FS domain therefore represents a new integrin ligand. 19153605 1 51 theme T-cell 204:209 arg1 priming 211:217 efficient T-cell priming 194:217 efficient T-cell priming by dendritic cells 194:236 Mindin (spondin-2) is an extracellular matrix protein of unknown structure that is required for efficient T-cell priming by dendritic cells. 19153605 0 52 theme integrin 48:55 arg1 ligand 57:62 integrin ligand 48:62 integrin ligand 48:62 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 19153605 10 53 theme adaptive 1208:1215 arg1 responses 1235:1243 both adaptive and innate immune responses 1203:1243 both adaptive and innate immune responses 1203:1243 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 10 54 theme TSR 1159:1161 arg1 mindin 1174:1179 mindin 1174:1179 mindin 1174:1179 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 10 54 theme TSR 1159:1161 arg1 domains 1163:1169 the FS and TSR domains 1148:1169 the FS and TSR domains of mindin 1148:1179 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 10 55 theme mindin 1174:1179 arg1 mindin 1174:1179 mindin 1174:1179 mindin 1174:1179 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 10 55 theme mindin 1174:1179 arg1 domains 1163:1169 the FS and TSR domains 1148:1169 the FS and TSR domains of mindin 1148:1179 Through these dual interactions, the FS and TSR domains of mindin promote activation of both adaptive and innate immune responses. 19153605 0 56 theme pattern 68:74 arg1 recognition 76:86 pattern recognition 68:86 pattern recognition 68:86 Structure of the F-spondin domain of mindin, an integrin ligand and pattern recognition molecule. 17614963 3 0 theme acid 620:623 arg1 residues 625:632 189 amino acid residues 610:632 189 amino acid residues 610:632 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 7 1 theme fibronectin 1326:1336 arg1 matrix 1354:1359 fibronectin comprising bone matrix 1326:1359 fibronectin comprising bone matrix 1326:1359 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 3 2 theme pediatric 429:437 arg1 urine 439:443 pediatric urine 429:443 pediatric urine 429:443 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 1 3 theme age 245:247 arg1 years 236:240 18 years 233:240 18 years of age 233:247 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 4 4 theme COS-7 851:855 arg1 cells 857:861 COS-7 cells 851:861 COS-7 cells transfected with Y1- or Y2-expressing vectors 851:907 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 7 5 theme matrix 1354:1359 arg1 degradation 1311:1321 degradation 1311:1321 degradation of fibronectin comprising bone matrix 1311:1359 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 8 6 theme juvenile-specific 1508:1524 arg1 fragments 1538:1546 juvenile-specific fibronectin fragments 1508:1546 juvenile-specific fibronectin fragments excreted into urine 1508:1566 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 6 7 theme osteoclast 1235:1244 arg1 cells 1246:1250 osteoclast cells 1235:1250 osteoclast cells 1235:1250 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 8 8 theme first 1449:1453 arg1 report 1455:1460 the first report 1445:1460 the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine 1445:1566 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 8 8 theme first 1449:1453 arg1 This 1437:1440 This 1437:1440 This 1437:1440 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 7 9 from children 1411:1418 arg1 process 1372:1378 the process 1368:1378 the process of vigorous bone resorption in children and adolescents 1368:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 7 10 theme comprising 1338:1347 arg1 matrix 1354:1359 fibronectin comprising bone matrix 1326:1359 fibronectin comprising bone matrix 1326:1359 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 3 11 from R911 679:682 arg1 region 659:664 the region 655:664 the region from A723 to R911 of fibronectin 655:697 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 8 12 theme fragments 1538:1546 arg1 characterization 1488:1503 characterization 1488:1503 characterization 1488:1503 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 8 12 theme fragments 1538:1546 arg1 identification 1469:1482 identification 1469:1482 identification 1469:1482 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 2 13 theme Ugl-Y 283:287 arg1 analysis 271:278 Isoelectric focusing analysis 250:278 Isoelectric focusing analysis of Ugl-Y 250:287 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 7 14 from process 1372:1378 arg1 children 1411:1418 children 1411:1418 children 1411:1418 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 7 14 from process 1372:1378 arg1 adolescents 1424:1434 adolescents 1424:1434 adolescents 1424:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 3 15 dep consists 598:605 arg1 whereas 700:706 whereas 700:706 whereas 700:706 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 4 16 theme lysate 830:835 arg1 analysis 814:821 Electrophoretic analysis 798:821 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors 798:907 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 7 17 theme bone 1349:1352 arg1 matrix 1354:1359 fibronectin comprising bone matrix 1326:1359 fibronectin comprising bone matrix 1326:1359 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 3 18 theme complete 376:383 arg1 structure 385:393 the complete structure 372:393 the complete structure of Ugl-Y 372:402 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 7 19 from adolescents 1424:1434 arg1 process 1372:1378 the process 1368:1378 the process of vigorous bone resorption in children and adolescents 1368:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 5 20 theme western 1072:1078 arg1 blotting 1080:1087 western blotting 1072:1087 western blotting 1072:1087 Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. 17614963 6 21 dep in 1169:1170 arg1 vitro 1172:1176 vitro 1172:1176 vitro 1172:1176 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 2 22 theme pH 339:340 arg1 region 342:347 the pH region 335:347 the pH region around 3 335:356 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 3 23 dep R911 679:682 arg1 to 676:677 to 676:677 to 676:677 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 8 24 from report 1455:1460 arg1 characterization 1488:1503 characterization 1488:1503 characterization 1488:1503 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 8 24 from report 1455:1460 arg1 identification 1469:1482 identification 1469:1482 identification 1469:1482 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 4 25 theme sequences 998:1006 arg1 validity 982:989 the validity 978:989 the validity of the sequences determined 978:1017 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 2 26 dep focusing 262:269 arg1 Isoelectric 250:260 Isoelectric 250:260 Isoelectric 250:260 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 4 27 theme Y2-expressing 887:899 arg1 vectors 901:907 Y1- or Y2-expressing vectors 880:907 Y1- or Y2-expressing vectors 880:907 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 5 28 theme pediatric 1044:1052 arg1 serum 1054:1058 pediatric serum 1044:1058 pediatric serum 1044:1058 Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. 17614963 1 29 theme young 94:98 arg1 glycoprotein 120:131 young age-related urinary glycoprotein 94:131 young age-related urinary glycoprotein 94:131 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 1 29 theme young 94:98 arg1 Ugl-Y 87:91 Ugl-Y 87:91 Ugl-Y (young age-related urinary glycoprotein) 87:132 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 6 30 theme acid 1206:1209 arg1 protease 1211:1218 acid protease 1206:1218 acid protease in extracts of osteoclast cells 1206:1250 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 0 31 theme fibronectin 24:34 arg1 fragments 36:44 novel fibronectin fragments 18:44 novel fibronectin fragments 18:44 Identification of novel fibronectin fragments detected specifically in juvenile urine. 17614963 5 32 theme serum 1054:1058 arg1 purification 1028:1039 Partial purification 1020:1039 Partial purification of pediatric serum followed by western blotting 1020:1087 Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. 17614963 8 33 dep identification 1469:1482 arg1 the 1465:1467 the 1465:1467 the 1465:1467 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 5 34 attach derived 1112:1118 arg1 plasma 1125:1130 plasma 1125:1130 plasma 1125:1130 Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. 17614963 5 34 attach derived 1112:1118 arg2 Ugl-Y 1103:1107 Ugl-Y 1103:1107 Ugl-Y 1103:1107 Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. 17614963 1 35 theme age-related 100:110 arg1 glycoprotein 120:131 young age-related urinary glycoprotein 94:131 young age-related urinary glycoprotein 94:131 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 1 35 theme age-related 100:110 arg1 Ugl-Y 87:91 Ugl-Y 87:91 Ugl-Y (young age-related urinary glycoprotein) 87:132 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 0 36 theme novel 18:22 arg1 fragments 36:44 novel fibronectin fragments 18:44 novel fibronectin fragments 18:44 Identification of novel fibronectin fragments detected specifically in juvenile urine. 17614963 6 37 from protease 1211:1218 arg1 extracts 1223:1230 extracts 1223:1230 extracts of osteoclast cells 1223:1250 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 1 38 gly glycoprotein 120:131 arg1 glycoprotein 120:131 young age-related urinary glycoprotein 94:131 young age-related urinary glycoprotein 94:131 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 1 38 gly glycoprotein 120:131 arg1 Ugl-Y 87:91 Ugl-Y 87:91 Ugl-Y (young age-related urinary glycoprotein) 87:132 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 1 39 theme urinary 112:118 arg1 glycoprotein 120:131 young age-related urinary glycoprotein 94:131 young age-related urinary glycoprotein 94:131 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 1 39 theme urinary 112:118 arg1 Ugl-Y 87:91 Ugl-Y 87:91 Ugl-Y (young age-related urinary glycoprotein) 87:132 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 4 40 theme Electrophoretic 798:812 arg1 analysis 814:821 Electrophoretic analysis 798:821 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors 798:907 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 8 41 theme fibronectin 1526:1536 arg1 fragments 1538:1546 juvenile-specific fibronectin fragments 1508:1546 juvenile-specific fibronectin fragments excreted into urine 1508:1566 This is the first report on the identification and characterization of juvenile-specific fibronectin fragments excreted into urine. 17614963 4 42 theme specific 914:921 arg1 bands 923:927 specific bands 914:927 specific bands corresponding to Y1 or Y2, respectively 914:967 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 1 43 theme healthy 210:216 arg1 subjects 218:225 healthy subjects 210:225 healthy subjects 210:225 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 0 44 theme fragments 36:44 arg1 Identification 0:13 Identification 0:13 Identification of novel fibronectin fragments 0:44 Identification of novel fibronectin fragments detected specifically in juvenile urine. 17614963 6 45 theme fibronectin 1191:1201 arg1 digestion 1178:1186 in vitro digestion 1169:1186 in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells 1169:1250 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 3 46 from R903 792:795 arg1 region 772:777 the region 768:777 the region from A723 to R903 768:795 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 3 47 theme Ugl-Y 398:402 arg1 structure 385:393 the complete structure 372:393 the complete structure of Ugl-Y 372:402 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 5 48 theme Partial 1020:1026 arg1 purification 1028:1039 Partial purification 1020:1039 Partial purification of pediatric serum followed by western blotting 1020:1087 Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. 17614963 1 49 from subjects 218:225 arg1 urine 199:203 urine 199:203 urine from healthy subjects under 18 years of age 199:247 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 4 50 theme Y1- 880:882 arg1 vectors 901:907 Y1- or Y2-expressing vectors 880:907 Y1- or Y2-expressing vectors 880:907 Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2, respectively, showing the validity of the sequences determined. 17614963 7 51 theme bone 1392:1395 arg1 resorption 1397:1406 vigorous bone resorption 1383:1406 vigorous bone resorption in children and adolescents 1383:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 3 52 theme protein 517:523 arg1 sequencing 525:534 protein sequencing 517:534 protein sequencing 517:534 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 7 53 from resorption 1397:1406 arg1 children 1411:1418 children 1411:1418 children 1411:1418 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 7 53 from resorption 1397:1406 arg1 adolescents 1424:1434 adolescents 1424:1434 adolescents 1424:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 7 54 theme resorption 1397:1406 arg1 process 1372:1378 the process 1368:1378 the process of vigorous bone resorption in children and adolescents 1368:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 2 55 theme focusing 262:269 arg1 analysis 271:278 Isoelectric focusing analysis 250:278 Isoelectric focusing analysis of Ugl-Y 250:287 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 3 56 theme resulting 481:489 arg1 peptides 491:498 the resulting peptides 477:498 the resulting peptides 477:498 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 6 57 theme in 1169:1170 arg1 digestion 1178:1186 in vitro digestion 1169:1186 in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells 1169:1250 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 2 58 theme bands 310:314 arg1 Y1 317:318 Y1 317:318 Y1 317:318 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 2 58 theme bands 310:314 arg1 set 297:299 a set 295:299 a set of three bands, Y1, Y2 and Y3, 295:330 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 2 58 theme bands 310:314 arg1 bands 310:314 three bands 304:314 three bands 304:314 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 2 58 theme bands 310:314 arg1 Y3 328:329 Y3 328:329 Y3 328:329 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 2 58 theme bands 310:314 arg1 Y2 321:322 Y2 321:322 Y2 321:322 Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. 17614963 3 59 theme amino 727:731 arg1 residues 738:745 181 amino acid residues 723:745 181 amino acid residues 723:745 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 7 60 theme vigorous 1383:1390 arg1 resorption 1397:1406 vigorous bone resorption 1383:1406 vigorous bone resorption in children and adolescents 1383:1434 These findings suggest that Ugl-Y is probably produced by degradation of fibronectin comprising bone matrix during the process of vigorous bone resorption in children and adolescents. 17614963 6 61 theme cells 1246:1250 arg1 extracts 1223:1230 extracts 1223:1230 extracts of osteoclast cells 1223:1250 Furthermore, Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. 17614963 3 62 dep R903 792:795 arg1 to 789:790 to 789:790 to 789:790 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 1 63 theme age-specific 140:151 arg1 protein 153:159 an age-specific protein 137:159 an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age 137:247 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 1 63 theme age-specific 140:151 arg1 Ugl-Y 87:91 Ugl-Y 87:91 Ugl-Y (young age-related urinary glycoprotein) 87:132 Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. 17614963 3 64 theme acid 733:736 arg1 residues 738:745 181 amino acid residues 723:745 181 amino acid residues 723:745 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 0 65 theme juvenile 71:78 arg1 urine 80:84 juvenile urine 71:84 juvenile urine 71:84 Identification of novel fibronectin fragments detected specifically in juvenile urine. 17614963 3 66 theme amino 614:618 arg1 residues 625:632 189 amino acid residues 610:632 189 amino acid residues 610:632 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 3 67 theme fibronectin 687:697 arg1 fibronectin 687:697 fibronectin 687:697 fibronectin 687:697 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 17614963 3 67 theme fibronectin 687:697 arg1 R911 679:682 R911 679:682 R911 679:682 To determine the complete structure of Ugl-Y, purified Y1 and Y2 from pediatric urine were enzymatically cleaved, and the resulting peptides were analyzed by protein sequencing and/or MALDI-TOF MS. As a result, it was demonstrated that Y1 consists of 189 amino acid residues, and is identical to the region from A723 to R911 of fibronectin, whereas Y2 consists of 181 amino acid residues, and is identical to the region from A723 to R903. 19119025 6 0 theme PLC 1050:1052 arg1 components 1054:1063 other defined PLC components 1036:1063 other defined PLC components 1036:1063 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 6 1 theme tapasin-ERp57 998:1010 arg1 structure 1012:1020 the tapasin-ERp57 structure 994:1020 the tapasin-ERp57 structure 994:1020 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 1 2 theme loading 155:161 arg1 class 202:206 loading major histocompatibility complex (MHC) class I 155:208 loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 155:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 0 3 theme thiol 82:86 arg1 heterodimer 103:113 the tapasin-ERp57 thiol oxidoreductase heterodimer 64:113 the tapasin-ERp57 thiol oxidoreductase heterodimer 64:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 3 4 theme tapasin-ERp57 523:535 arg1 core 537:540 the tapasin-ERp57 core 519:540 the tapasin-ERp57 core of the PLC 519:551 Here, we present the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC. 19119025 4 5 theme protein 713:719 arg1 isomerase 731:739 protein disulfide isomerase 713:739 a protein disulfide isomerase family member interacting with substrate 711:780 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 0 6 theme tapasin-ERp57 68:80 arg1 heterodimer 103:113 the tapasin-ERp57 thiol oxidoreductase heterodimer 64:113 the tapasin-ERp57 thiol oxidoreductase heterodimer 64:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 3 7 theme core 537:540 arg1 structure 506:514 the 2.6 A resolution structure 485:514 the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC 485:551 Here, we present the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC. 19119025 2 8 theme peptide-loading 284:298 arg1 PLC 309:311 PLC 309:311 PLC 309:311 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 2 8 theme peptide-loading 284:298 arg1 complex 300:306 the multimeric peptide-loading complex 269:306 the multimeric peptide-loading complex (PLC) 269:312 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 0 9 theme heterodimer 103:113 arg1 structure 51:59 the structure 47:59 the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer 47:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 4 10 theme ERp57 610:614 arg1 domains 626:632 both ERp57 catalytic domains 605:632 both ERp57 catalytic domains 605:632 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 2 11 theme multimeric 273:282 arg1 PLC 309:311 PLC 309:311 PLC 309:311 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 2 11 theme multimeric 273:282 arg1 complex 300:306 the multimeric peptide-loading complex 269:306 the multimeric peptide-loading complex (PLC) 269:312 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 5 12 theme MHC 866:868 arg1 molecules 878:886 MHC class I molecules 866:886 MHC class I molecules 866:886 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 1 13 theme major 163:167 arg1 class 202:206 loading major histocompatibility complex (MHC) class I 155:208 loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 155:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 0 14 theme oxidoreductase 88:101 arg1 heterodimer 103:113 the tapasin-ERp57 thiol oxidoreductase heterodimer 64:113 the tapasin-ERp57 thiol oxidoreductase heterodimer 64:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 4 15 theme member 748:753 arg1 example 700:706 an example 697:706 an example of a protein disulfide isomerase family member interacting with substrate 697:780 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 6 16 theme molecular 1079:1087 arg1 model 1089:1093 a molecular model 1077:1093 a molecular model that illuminates the processes involved in MHC class I peptide loading 1077:1164 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 1 17 theme histocompatibility 169:186 arg1 class 202:206 loading major histocompatibility complex (MHC) class I 155:208 loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 155:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 2 18 theme optimal 446:452 arg1 activity 458:465 optimal PLC activity 446:465 optimal PLC activity 446:465 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 6 19 theme peptide 1150:1156 arg1 loading 1158:1164 MHC class I peptide loading 1138:1164 MHC class I peptide loading 1138:1164 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 2 20 dep disulfide-linked 319:334 arg1 stable 337:342 stable 337:342 stable 337:342 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 1 21 theme complex 188:194 arg1 class 202:206 loading major histocompatibility complex (MHC) class I 155:208 loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 155:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 5 22 theme editing 929:935 arg1 functions 937:945 editing functions 929:945 editing functions 929:945 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 2 23 theme oxidoreductase 371:384 arg1 ERp57 386:390 the thiol oxidoreductase ERp57 361:390 the thiol oxidoreductase ERp57 361:390 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 0 24 theme class 18:22 arg1 loading 34:40 MHC class I peptide loading 14:40 MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer 14:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 6 25 theme MHC 1138:1140 arg1 class 1142:1146 MHC class I 1138:1148 MHC class I peptide loading 1138:1164 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 4 26 theme disulfide 721:729 arg1 isomerase 731:739 protein disulfide isomerase 713:739 a protein disulfide isomerase family member interacting with substrate 711:780 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 1 27 theme MHC 197:199 arg1 class 202:206 loading major histocompatibility complex (MHC) class I 155:208 loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 155:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 0 28 theme MHC 14:16 arg1 class 18:22 MHC class I 14:24 MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer 14:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 5 29 theme heterodimer 968:978 arg1 functions 937:945 editing functions 929:945 editing functions 929:945 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 5 29 theme heterodimer 968:978 arg1 loading 917:923 peptide loading 909:923 peptide loading 909:923 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 3 30 theme A 493:493 arg1 structure 506:514 the 2.6 A resolution structure 485:514 the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC 485:551 Here, we present the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC. 19119025 3 31 theme resolution 495:504 arg1 structure 506:514 the 2.6 A resolution structure 485:514 the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC 485:551 Here, we present the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC. 19119025 0 32 theme peptide 26:32 arg1 loading 34:40 MHC class I peptide loading 14:40 MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer 14:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 2 33 theme covalent 402:409 arg1 interaction 411:421 this covalent interaction 397:421 this covalent interaction 397:421 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 5 34 theme class 870:874 arg1 molecules 878:886 MHC class I molecules 866:886 MHC class I molecules 866:886 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 1 35 theme class 202:206 arg1 molecules 210:218 loading major histocompatibility complex (MHC) class I molecules 155:218 loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 155:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 0 36 from structure 51:59 arg1 loading 34:40 MHC class I peptide loading 14:40 MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer 14:113 Insights into MHC class I peptide loading from the structure of the tapasin-ERp57 thiol oxidoreductase heterodimer. 19119025 6 37 theme other 1036:1040 arg1 components 1054:1063 other defined PLC components 1036:1063 other defined PLC components 1036:1063 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 5 38 theme I 876:876 arg1 molecules 878:886 MHC class I molecules 866:886 MHC class I molecules 866:886 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 5 39 theme Mutational 783:792 arg1 analysis 794:801 Mutational analysis 783:801 Mutational analysis 783:801 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 4 40 theme isomerase 731:739 arg1 member 748:753 a protein disulfide isomerase family member 711:753 a protein disulfide isomerase family member interacting with substrate 711:780 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 5 41 theme tapasin-ERp57 954:966 arg1 heterodimer 968:978 the tapasin-ERp57 heterodimer 950:978 the tapasin-ERp57 heterodimer 950:978 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 2 42 theme PLC 454:456 arg1 activity 458:465 optimal PLC activity 446:465 optimal PLC activity 446:465 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 1 43 gly glycoprotein 129:140 arg1 Tapasin 116:122 Tapasin 116:122 Tapasin 116:122 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 1 43 gly glycoprotein 129:140 arg1 glycoprotein 129:140 a glycoprotein 127:140 a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 127:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 6 44 theme class 1142:1146 arg1 loading 1158:1164 MHC class I peptide loading 1138:1164 MHC class I peptide loading 1138:1164 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 19119025 5 45 from surface 826:832 arg1 tapasin 837:843 tapasin 837:843 tapasin 837:843 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 3 46 theme PLC 549:551 arg1 core 537:540 the tapasin-ERp57 core 519:540 the tapasin-ERp57 core of the PLC 519:551 Here, we present the 2.6 A resolution structure of the tapasin-ERp57 core of the PLC. 19119025 2 47 with heterodimer 344:354 arg1 ERp57 386:390 the thiol oxidoreductase ERp57 361:390 the thiol oxidoreductase ERp57 361:390 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 2 48 theme disulfide-linked 319:334 arg1 It 249:250 It 249:250 It 249:250 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 2 48 theme disulfide-linked 319:334 arg1 heterodimer 344:354 a disulfide-linked, stable heterodimer 317:354 a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57 317:390 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 4 49 theme heterodimer 671:681 arg1 stability 654:662 the stability 650:662 the stability of the heterodimer 650:681 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 4 50 theme family 741:746 arg1 member 748:753 a protein disulfide isomerase family member 711:753 a protein disulfide isomerase family member interacting with substrate 711:780 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 5 51 theme peptide 909:915 arg1 loading 917:923 peptide loading 909:923 peptide loading 909:923 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 4 52 theme catalytic 616:624 arg1 domains 626:632 both ERp57 catalytic domains 605:632 both ERp57 catalytic domains 605:632 The structure revealed that tapasin interacts with both ERp57 catalytic domains, accounting for the stability of the heterodimer, and provided an example of a protein disulfide isomerase family member interacting with substrate. 19119025 2 53 theme thiol 365:369 arg1 ERp57 386:390 the thiol oxidoreductase ERp57 361:390 the thiol oxidoreductase ERp57 361:390 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 1 54 theme high-affinity 225:237 arg1 peptides 239:246 high-affinity peptides 225:246 high-affinity peptides 225:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 2 55 link disulfide-linked 319:334 arg1 It 249:250 It 249:250 It 249:250 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 2 55 link disulfide-linked 319:334 arg1 heterodimer 344:354 a disulfide-linked, stable heterodimer 317:354 a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57 317:390 It functions within the multimeric peptide-loading complex (PLC) as a disulfide-linked, stable heterodimer with the thiol oxidoreductase ERp57, and this covalent interaction is required to support optimal PLC activity. 19119025 5 56 theme conserved 816:824 arg1 surface 826:832 a conserved surface 814:832 a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer 814:978 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 5 56 theme conserved 816:824 arg1 critical 896:903 critical 896:903 critical 896:903 Mutational analysis identified a conserved surface on tapasin that interacted with MHC class I molecules and was critical for peptide loading and editing functions of the tapasin-ERp57 heterodimer. 19119025 1 57 with molecules 210:218 arg1 peptides 239:246 high-affinity peptides 225:246 high-affinity peptides 225:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 1 58 theme critical 142:149 arg1 Tapasin 116:122 Tapasin 116:122 Tapasin 116:122 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 1 58 theme critical 142:149 arg1 glycoprotein 129:140 a glycoprotein 127:140 a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides 127:246 Tapasin is a glycoprotein critical for loading major histocompatibility complex (MHC) class I molecules with high-affinity peptides. 19119025 6 59 theme defined 1042:1048 arg1 components 1054:1063 other defined PLC components 1036:1063 other defined PLC components 1036:1063 By combining the tapasin-ERp57 structure with those of other defined PLC components, we present a molecular model that illuminates the processes involved in MHC class I peptide loading. 18093521 1 0 theme postsynaptic 150:161 arg1 proteins 177:184 postsynaptic cell adhesion proteins 150:184 postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis 150:260 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 1 0 theme postsynaptic 150:161 arg1 neuroligins 134:144 The neuroligins 130:144 The neuroligins 130:144 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 4 1 dep involved 1044:1051 arg1 interactions 1023:1034 the Ca2+-dependent interactions 1004:1034 the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association 1004:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 1 dep involved 1044:1051 arg1 network 1036:1042 network 1036:1042 network 1036:1042 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 0 2 theme complex 77:83 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex 0:83 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 3 3 dep electronegative 768:782 arg1 hydrophilic 785:795 hydrophilic 785:795 hydrophilic 785:795 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 4 theme neuroligin 537:546 arg1 dimer 548:552 the functional neuroligin dimer 522:552 the functional neuroligin dimer 522:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 4 5 theme limited 948:954 arg1 resolution 956:965 a limited resolution 946:965 a limited resolution 946:965 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 6 theme neurexin 924:931 arg1 orientation 899:909 the precise orientation 887:909 the precise orientation of the bound neurexin 887:931 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 7 theme complex 871:877 arg1 structure 831:839 The structure 827:839 The structure of a beta-neurexin-neuroligin complex 827:877 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 8 from information 989:999 arg1 interactions 1023:1034 the Ca2+-dependent interactions 1004:1034 the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association 1004:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 8 from information 989:999 arg1 network 1036:1042 network 1036:1042 network 1036:1042 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 9 theme substantial 977:987 arg1 information 989:999 substantial information 977:999 substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association 977:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 10 theme bound 918:922 arg1 neurexin 924:931 the bound neurexin 914:931 the bound neurexin 914:931 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 3 11 theme surface 576:582 arg1 loops 584:588 surface loops 576:588 surface loops surrounding the vestigial active center 576:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 1 12 theme adhesion 168:175 arg1 proteins 177:184 postsynaptic cell adhesion proteins 150:184 postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis 150:260 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 1 12 theme adhesion 168:175 arg1 neuroligins 134:144 The neuroligins 130:144 The neuroligins 130:144 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 3 13 theme crystal 374:380 arg1 structure 382:390 The crystal structure 370:390 The crystal structure of a neuroligin 370:406 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 14 theme macromolecular 727:740 arg1 dipole 742:747 a macromolecular dipole 725:747 a macromolecular dipole 725:747 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 4 15 theme trans-synaptic 1056:1069 arg1 association 1091:1101 trans-synaptic neurexin-neuroligin association 1056:1101 trans-synaptic neurexin-neuroligin association 1056:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 0 16 dep determinants 86:97 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex 0:83 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 2 17 from Mutations 263:271 arg1 genes 304:308 the neuroligin and neurexin genes 276:308 genes 304:308 Mutations in the neuroligin and neurexin genes appear to be associated with autism and mental retardation. 18093521 3 18 theme dipole 742:747 arg1 presence 753:760 presence 753:760 presence 753:760 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 18 theme dipole 742:747 arg1 features 416:423 features 416:423 features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer 416:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 18 theme dipole 742:747 arg1 conformations 559:571 the conformations 555:571 the conformations of surface loops surrounding the vestigial active center 555:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 18 theme dipole 742:747 arg1 location 635:642 the location 631:642 the location of determinants that are critical for folding and processing 631:703 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 18 theme dipole 742:747 arg1 absence 714:720 the absence 710:720 the absence of a macromolecular dipole 710:747 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 5 19 theme recognition 1282:1292 arg1 events 1294:1299 abnormal processing or recognition events 1259:1299 events 1294:1299 These structures exemplify how an alpha/beta-hydrolase fold varies in surface topography to confer adhesion properties and provide templates for analyzing abnormal processing or recognition events associated with autism. 18093521 0 20 theme Structural 0:9 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex 0:83 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 3 21 theme electronegative 768:782 arg1 surface 797:803 an electronegative, hydrophilic surface 765:803 an electronegative, hydrophilic surface for neurexin binding 765:824 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 4 22 theme precise 891:897 arg1 orientation 899:909 the precise orientation 887:909 the precise orientation of the bound neurexin 887:931 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 3 23 theme vestigial 606:614 arg1 center 623:628 the vestigial active center 602:628 the vestigial active center 602:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 24 theme active 456:461 arg1 relatives 463:471 its catalytically active relatives 438:471 its catalytically active relatives 438:471 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 24 theme active 456:461 arg1 interface 504:512 the fully hydrophobic interface 482:512 the fully hydrophobic interface forming the functional neuroligin dimer 482:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 25 theme loops 584:588 arg1 presence 753:760 presence 753:760 presence 753:760 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 25 theme loops 584:588 arg1 features 416:423 features 416:423 features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer 416:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 25 theme loops 584:588 arg1 conformations 559:571 the conformations 555:571 the conformations of surface loops surrounding the vestigial active center 555:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 25 theme loops 584:588 arg1 location 635:642 the location 631:642 the location of determinants that are critical for folding and processing 631:703 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 25 theme loops 584:588 arg1 absence 714:720 the absence 710:720 the absence of a macromolecular dipole 710:747 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 26 theme neuroligin 397:406 arg1 structure 382:390 The crystal structure 370:390 The crystal structure of a neuroligin 370:406 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 0 27 theme cell 115:118 arg1 adhesion 120:127 cell adhesion 115:127 cell adhesion 115:127 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 1 28 theme cell 163:166 arg1 proteins 177:184 postsynaptic cell adhesion proteins 150:184 postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis 150:260 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 1 28 theme cell 163:166 arg1 neuroligins 134:144 The neuroligins 130:144 The neuroligins 130:144 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 1 29 theme presynaptic 210:220 arg1 neurexins 222:230 presynaptic neurexins 210:230 presynaptic neurexins 210:230 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 0 30 theme protein 36:42 arg1 neuroligin 44:53 the synaptic protein neuroligin 23:53 the synaptic protein neuroligin 23:53 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 0 31 theme synaptic 27:34 arg1 neuroligin 44:53 the synaptic protein neuroligin 23:53 the synaptic protein neuroligin 23:53 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 4 32 theme neurexin-neuroligin 1071:1089 arg1 association 1091:1101 trans-synaptic neurexin-neuroligin association 1056:1101 trans-synaptic neurexin-neuroligin association 1056:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 3 33 located found 429:433 arg1 relatives 463:471 its catalytically active relatives 438:471 its catalytically active relatives 438:471 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 33 located found 429:433 arg2 features 416:423 features 416:423 features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer 416:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 33 located found 429:433 arg1 interface 504:512 the fully hydrophobic interface 482:512 the fully hydrophobic interface forming the functional neuroligin dimer 482:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 34 theme neurexin 809:816 arg1 binding 818:824 neurexin binding 809:824 neurexin binding 809:824 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 35 theme functional 526:535 arg1 dimer 548:552 the functional neuroligin dimer 522:552 the functional neuroligin dimer 522:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 36 theme active 616:621 arg1 center 623:628 the vestigial active center 602:628 the vestigial active center 602:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 5 37 theme adhesion 1203:1210 arg1 properties 1212:1221 adhesion properties 1203:1221 adhesion properties 1203:1221 These structures exemplify how an alpha/beta-hydrolase fold varies in surface topography to confer adhesion properties and provide templates for analyzing abnormal processing or recognition events associated with autism. 18093521 0 38 theme neuroligin 44:53 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex 0:83 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 1 39 with associations 192:203 arg1 neurexins 222:230 presynaptic neurexins 210:230 presynaptic neurexins 210:230 The neuroligins are postsynaptic cell adhesion proteins whose associations with presynaptic neurexins participate in synaptogenesis. 18093521 4 40 theme beta-neurexin-neuroligin 846:869 arg1 complex 871:877 a beta-neurexin-neuroligin complex 844:877 a beta-neurexin-neuroligin complex 844:877 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 5 41 theme surface 1174:1180 arg1 topography 1182:1191 surface topography 1174:1191 surface topography 1174:1191 These structures exemplify how an alpha/beta-hydrolase fold varies in surface topography to confer adhesion properties and provide templates for analyzing abnormal processing or recognition events associated with autism. 18093521 3 42 theme surface 797:803 arg1 presence 753:760 presence 753:760 presence 753:760 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 42 theme surface 797:803 arg1 features 416:423 features 416:423 features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer 416:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 42 theme surface 797:803 arg1 conformations 559:571 the conformations 555:571 the conformations of surface loops surrounding the vestigial active center 555:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 42 theme surface 797:803 arg1 location 635:642 the location 631:642 the location of determinants that are critical for folding and processing 631:703 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 42 theme surface 797:803 arg1 absence 714:720 the absence 710:720 the absence of a macromolecular dipole 710:747 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 43 theme determinants 647:658 arg1 presence 753:760 presence 753:760 presence 753:760 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 43 theme determinants 647:658 arg1 features 416:423 features 416:423 features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer 416:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 43 theme determinants 647:658 arg1 conformations 559:571 the conformations 555:571 the conformations of surface loops surrounding the vestigial active center 555:628 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 43 theme determinants 647:658 arg1 location 635:642 the location 631:642 the location of determinants that are critical for folding and processing 631:703 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 3 43 theme determinants 647:658 arg1 absence 714:720 the absence 710:720 the absence of a macromolecular dipole 710:747 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 2 44 theme mental 350:355 arg1 retardation 357:367 mental retardation 350:367 mental retardation 350:367 Mutations in the neuroligin and neurexin genes appear to be associated with autism and mental retardation. 18093521 2 45 theme neurexin 295:302 arg1 genes 304:308 the neuroligin and neurexin genes 276:308 genes 304:308 Mutations in the neuroligin and neurexin genes appear to be associated with autism and mental retardation. 18093521 5 46 theme alpha/beta-hydrolase 1138:1157 arg1 fold 1159:1162 an alpha/beta-hydrolase fold 1135:1162 an alpha/beta-hydrolase fold 1135:1162 These structures exemplify how an alpha/beta-hydrolase fold varies in surface topography to confer adhesion properties and provide templates for analyzing abnormal processing or recognition events associated with autism. 18093521 2 47 theme neuroligin 280:289 arg1 genes 304:308 the neuroligin and neurexin genes 276:308 genes 304:308 Mutations in the neuroligin and neurexin genes appear to be associated with autism and mental retardation. 18093521 3 48 theme hydrophobic 492:502 arg1 interface 504:512 the fully hydrophobic interface 482:512 the fully hydrophobic interface forming the functional neuroligin dimer 482:552 The crystal structure of a neuroligin reveals features not found in its catalytically active relatives, such as the fully hydrophobic interface forming the functional neuroligin dimer; the conformations of surface loops surrounding the vestigial active center; the location of determinants that are critical for folding and processing; and the absence of a macromolecular dipole and presence of an electronegative, hydrophilic surface for neurexin binding. 18093521 0 49 theme beta-neurexin 63:75 arg1 complex 77:83 its beta-neurexin complex 59:83 its beta-neurexin complex 59:83 Structural analysis of the synaptic protein neuroligin and its beta-neurexin complex: determinants for folding and cell adhesion. 18093521 4 50 theme Ca2+-dependent 1008:1021 arg1 interactions 1023:1034 the Ca2+-dependent interactions 1004:1034 the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association 1004:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 50 theme Ca2+-dependent 1008:1021 arg1 network 1036:1042 network 1036:1042 network 1036:1042 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 4 51 dep reveals 879:885 arg1 provides 968:975 provides 968:975 provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association 968:1101 The structure of a beta-neurexin-neuroligin complex reveals the precise orientation of the bound neurexin and, despite a limited resolution, provides substantial information on the Ca2+-dependent interactions network involved in trans-synaptic neurexin-neuroligin association. 18093521 5 52 theme abnormal 1259:1266 arg1 processing 1268:1277 abnormal processing or recognition events 1259:1299 processing 1268:1277 These structures exemplify how an alpha/beta-hydrolase fold varies in surface topography to confer adhesion properties and provide templates for analyzing abnormal processing or recognition events associated with autism. 10209036 1 0 theme low 151:153 arg1 proteins 194:201 several low molecular weight transmembrane adaptor proteins 143:201 several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs 143:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 1 1 theme domain-containing 235:251 arg1 molecules 267:275 src homology (SH)2 domain-containing intracellular molecules 216:275 src homology (SH)2 domain-containing intracellular molecules 216:275 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 0 2 theme disulfide-linked 62:77 arg1 dimer 79:83 a novel disulfide-linked dimer 54:83 a novel disulfide-linked dimer 54:83 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 2 theme disulfide-linked 62:77 arg1 protein 39:45 SHP2-interacting transmembrane adaptor protein 0:45 SHP2-interacting transmembrane adaptor protein (SIT) 0:51 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 1 3 theme molecular 155:163 arg1 proteins 194:201 several low molecular weight transmembrane adaptor proteins 143:201 several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs 143:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 5 4 from Overexpression 729:742 arg1 cells 761:765 Jurkat cells 754:765 Jurkat cells 754:765 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 5 theme intracellular 253:265 arg1 molecules 267:275 src homology (SH)2 domain-containing intracellular molecules 216:275 src homology (SH)2 domain-containing intracellular molecules 216:275 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 0 6 theme human 96:100 arg1 activation 109:118 human T cell activation 96:118 human T cell activation 96:118 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 5 7 theme T 1184:1184 arg1 activation 1191:1200 T cell activation 1184:1200 T cell activation 1184:1200 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 3 8 link disulfide-linked 453:468 arg1 glycoprotein 482:493 a disulfide-linked homodimeric glycoprotein 451:493 a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes 451:526 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 3 8 link disulfide-linked 453:468 arg1 SIT 444:446 SIT 444:446 SIT 444:446 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 5 9 theme SHP2 1021:1024 arg1 binding 1010:1016 binding 1010:1016 binding of SHP2 to SIT 1010:1031 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 3 10 gly glycoprotein 482:493 arg1 glycoprotein 482:493 a disulfide-linked homodimeric glycoprotein 451:493 a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes 451:526 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 3 10 gly glycoprotein 482:493 arg1 SIT 444:446 SIT 444:446 SIT 444:446 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 5 11 theme cell 783:786 arg1 receptor- 788:796 T cell receptor- 781:796 T cell receptor- 781:796 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 4 12 theme phosphatase 657:667 arg1 SHP2 669:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 5 13 dep C. 998:999 arg1 required 1040:1047 required 1040:1047 required for inhibition of NF-AT induction 1040:1081 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 14 theme signaling 912:920 arg1 processes 922:930 signaling processes 912:930 signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2 912:1215 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 0 15 theme cell 104:107 arg1 activation 109:118 human T cell activation 96:118 human T cell activation 96:118 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 3 16 theme homodimeric 470:480 arg1 glycoprotein 482:493 a disulfide-linked homodimeric glycoprotein 451:493 a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes 451:526 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 3 16 theme homodimeric 470:480 arg1 SIT 444:446 SIT 444:446 SIT 444:446 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 5 17 theme phospholipase 984:996 arg1 C. 998:999 phospholipase C. 984:999 phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2 984:1215 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 18 theme weight 165:170 arg1 proteins 194:201 several low molecular weight transmembrane adaptor proteins 143:201 several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs 143:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 0 19 theme T 102:102 arg1 activation 109:118 human T cell activation 96:118 human T cell activation 96:118 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 5 20 theme T 876:876 arg1 cells 878:882 the nuclear factor of activated T cells 844:882 the nuclear factor of activated T cells (NF-AT) 844:890 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 20 theme T 876:876 arg1 NF-AT 885:889 NF-AT 885:889 NF-AT 885:889 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 21 theme cells 878:882 arg1 receptor- 788:796 T cell receptor- 781:796 T cell receptor- 781:796 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 21 theme cells 878:882 arg1 activation 830:839 phytohemagglutinin-mediated activation 802:839 phytohemagglutinin-mediated activation 802:839 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 22 theme transmembrane 172:184 arg1 proteins 194:201 several low molecular weight transmembrane adaptor proteins 143:201 several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs 143:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 0 23 theme transmembrane 17:29 arg1 SIT 48:50 SIT 48:50 SIT 48:50 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 23 theme transmembrane 17:29 arg1 protein 39:45 SHP2-interacting transmembrane adaptor protein 0:45 SHP2-interacting transmembrane adaptor protein (SIT) 0:51 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 23 theme transmembrane 17:29 arg1 dimer 79:83 a novel disulfide-linked dimer 54:83 a novel disulfide-linked dimer 54:83 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 5 24 theme NF-AT 1156:1160 arg1 pathways 1172:1179 NF-AT unrelated pathways 1156:1179 NF-AT unrelated pathways of T cell activation involving SHP2 1156:1215 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 25 theme phytohemagglutinin-mediated 802:828 arg1 activation 830:839 phytohemagglutinin-mediated activation 802:839 phytohemagglutinin-mediated activation 802:839 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 26 theme adaptor 186:192 arg1 proteins 194:201 several low molecular weight transmembrane adaptor proteins 143:201 several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs 143:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 0 27 theme SHP2-interacting 0:15 arg1 SIT 48:50 SIT 48:50 SIT 48:50 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 27 theme SHP2-interacting 0:15 arg1 protein 39:45 SHP2-interacting transmembrane adaptor protein 0:45 SHP2-interacting transmembrane adaptor protein (SIT) 0:51 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 27 theme SHP2-interacting 0:15 arg1 dimer 79:83 a novel disulfide-linked dimer 54:83 a novel disulfide-linked dimer 54:83 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 5 28 theme T 781:781 arg1 receptor- 788:796 T cell receptor- 781:796 T cell receptor- 781:796 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 2 29 theme group 377:381 arg1 molecule 360:367 a novel molecule 352:367 a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein) 352:441 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 1 30 theme cell 284:287 arg1 membrane 289:296 the cell membrane 280:296 the cell membrane 280:296 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 2 31 theme adaptor 426:432 arg1 SIT 390:392 SIT 390:392 SIT (SHP2 interacting transmembrane adaptor protein) 390:441 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 2 31 theme adaptor 426:432 arg1 protein 434:440 SHP2 interacting transmembrane adaptor protein 395:440 SHP2 interacting transmembrane adaptor protein 395:440 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 0 32 theme adaptor 31:37 arg1 SIT 48:50 SIT 48:50 SIT 48:50 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 32 theme adaptor 31:37 arg1 protein 39:45 SHP2-interacting transmembrane adaptor protein 0:45 SHP2-interacting transmembrane adaptor protein (SIT) 0:51 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 32 theme adaptor 31:37 arg1 dimer 79:83 a novel disulfide-linked dimer 54:83 a novel disulfide-linked dimer 54:83 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 4 33 theme protein 584:590 arg1 kinases 601:607 syk protein tyrosine kinases 580:607 syk protein tyrosine kinases 580:607 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 5 34 theme Jurkat 754:759 arg1 cells 761:765 Jurkat cells 754:765 Jurkat cells 754:765 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 2 35 theme novel 354:358 arg1 molecule 360:367 a novel molecule 352:367 a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein) 352:441 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 2 36 theme transmembrane 412:424 arg1 SIT 390:392 SIT 390:392 SIT (SHP2 interacting transmembrane adaptor protein) 390:441 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 2 36 theme transmembrane 412:424 arg1 protein 434:440 SHP2 interacting transmembrane adaptor protein 395:440 SHP2 interacting transmembrane adaptor protein 395:440 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 5 37 theme nuclear 848:854 arg1 cells 878:882 the nuclear factor of activated T cells 844:882 the nuclear factor of activated T cells (NF-AT) 844:890 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 37 theme nuclear 848:854 arg1 NF-AT 885:889 NF-AT 885:889 NF-AT 885:889 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 4 38 theme syk 580:582 arg1 kinases 601:607 syk protein tyrosine kinases 580:607 syk protein tyrosine kinases 580:607 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 2 39 theme interacting 400:410 arg1 SIT 390:392 SIT 390:392 SIT (SHP2 interacting transmembrane adaptor protein) 390:441 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 2 39 theme interacting 400:410 arg1 protein 434:440 SHP2 interacting transmembrane adaptor protein 395:440 SHP2 interacting transmembrane adaptor protein 395:440 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 4 40 theme inhibition 711:720 arg1 motif 722:726 an immunoreceptor tyrosine-based inhibition motif 678:726 an immunoreceptor tyrosine-based inhibition motif 678:726 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 1 41 theme src 216:218 arg1 molecules 267:275 src homology (SH)2 domain-containing intracellular molecules 216:275 src homology (SH)2 domain-containing intracellular molecules 216:275 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 4 42 theme tyrosine-based 696:709 arg1 motif 722:726 an immunoreceptor tyrosine-based inhibition motif 678:726 an immunoreceptor tyrosine-based inhibition motif 678:726 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 1 43 theme tyrosine-based 302:315 arg1 motifs 327:332 tyrosine-based signaling motifs 302:332 tyrosine-based signaling motifs 302:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 4 44 theme tyrosine 592:599 arg1 kinases 601:607 syk protein tyrosine kinases 580:607 syk protein tyrosine kinases 580:607 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 5 45 theme NF-AT 1123:1127 arg1 activity 1129:1136 NF-AT activity 1123:1136 NF-AT activity 1123:1136 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 46 theme NF-AT 1067:1071 arg1 induction 1073:1081 NF-AT induction 1067:1081 NF-AT induction 1067:1081 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 47 theme homology 220:227 arg1 molecules 267:275 src homology (SH)2 domain-containing intracellular molecules 216:275 src homology (SH)2 domain-containing intracellular molecules 216:275 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 5 48 theme of 863:864 arg1 cells 878:882 the nuclear factor of activated T cells 844:882 the nuclear factor of activated T cells (NF-AT) 844:890 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 48 theme of 863:864 arg1 NF-AT 885:889 NF-AT 885:889 NF-AT 885:889 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 49 theme signaling 317:325 arg1 motifs 327:332 tyrosine-based signaling motifs 302:332 tyrosine-based signaling motifs 302:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 5 50 theme induction 1073:1081 arg1 inhibition 1053:1062 inhibition 1053:1062 inhibition of NF-AT induction 1053:1081 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 0 51 link disulfide-linked 62:77 arg1 dimer 79:83 a novel disulfide-linked dimer 54:83 a novel disulfide-linked dimer 54:83 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 51 link disulfide-linked 62:77 arg1 protein 39:45 SHP2-interacting transmembrane adaptor protein 0:45 SHP2-interacting transmembrane adaptor protein (SIT) 0:51 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 5 52 theme activated 866:874 arg1 cells 878:882 the nuclear factor of activated T cells 844:882 the nuclear factor of activated T cells (NF-AT) 844:890 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 52 theme activated 866:874 arg1 NF-AT 885:889 NF-AT 885:889 NF-AT 885:889 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 53 theme unrelated 1162:1170 arg1 pathways 1172:1179 NF-AT unrelated pathways 1156:1179 NF-AT unrelated pathways of T cell activation involving SHP2 1156:1215 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 54 theme cell 1186:1189 arg1 activation 1191:1200 T cell activation 1184:1200 T cell activation 1184:1200 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 55 theme factor 856:861 arg1 cells 878:882 the nuclear factor of activated T cells 844:882 the nuclear factor of activated T cells (NF-AT) 844:890 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 55 theme factor 856:861 arg1 NF-AT 885:889 NF-AT 885:889 NF-AT 885:889 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 56 theme T 121:121 arg1 lymphocytes 123:133 T lymphocytes 121:133 T lymphocytes 121:133 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 4 57 theme SH2 626:628 arg1 SHP2 669:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 5 58 theme SIT 747:749 arg1 Overexpression 729:742 Overexpression 729:742 Overexpression of SIT in Jurkat cells 729:765 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 5 59 theme C. 998:999 arg1 activation 970:979 activation 970:979 activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2 970:1215 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 1 60 theme SH 230:231 arg1 molecules 267:275 src homology (SH)2 domain-containing intracellular molecules 216:275 src homology (SH)2 domain-containing intracellular molecules 216:275 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 0 61 theme novel 56:60 arg1 dimer 79:83 a novel disulfide-linked dimer 54:83 a novel disulfide-linked dimer 54:83 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 0 61 theme novel 56:60 arg1 protein 39:45 SHP2-interacting transmembrane adaptor protein 0:45 SHP2-interacting transmembrane adaptor protein (SIT) 0:51 SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. 10209036 2 62 theme SHP2 395:398 arg1 SIT 390:392 SIT 390:392 SIT (SHP2 interacting transmembrane adaptor protein) 390:441 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 2 62 theme SHP2 395:398 arg1 protein 434:440 SHP2 interacting transmembrane adaptor protein 395:440 SHP2 interacting transmembrane adaptor protein 395:440 We describe here a novel molecule of this group termed SIT (SHP2 interacting transmembrane adaptor protein). 10209036 3 63 theme disulfide-linked 453:468 arg1 glycoprotein 482:493 a disulfide-linked homodimeric glycoprotein 451:493 a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes 451:526 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 3 63 theme disulfide-linked 453:468 arg1 SIT 444:446 SIT 444:446 SIT 444:446 SIT is a disulfide-linked homodimeric glycoprotein that is expressed in lymphocytes. 10209036 4 64 theme immunoreceptor 681:694 arg1 motif 722:726 an immunoreceptor tyrosine-based inhibition motif 678:726 an immunoreceptor tyrosine-based inhibition motif 678:726 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 4 65 theme tyrosine 648:655 arg1 SHP2 669:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 5 66 theme activation 1191:1200 arg1 pathways 1172:1179 NF-AT unrelated pathways 1156:1179 NF-AT unrelated pathways of T cell activation involving SHP2 1156:1215 Overexpression of SIT in Jurkat cells downmodulates T cell receptor- and phytohemagglutinin-mediated activation of the nuclear factor of activated T cells (NF-AT) by interfering with signaling processes that are probably located upstream of activation of phospholipase C. However, binding of SHP2 to SIT is not required for inhibition of NF-AT induction, suggesting that SIT not only regulates NF-AT activity but also controls NF-AT unrelated pathways of T cell activation involving SHP2. 10209036 4 67 theme domain-containing 630:646 arg1 SHP2 669:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 the SH2 domain-containing tyrosine phosphatase SHP2 622:672 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 10209036 1 68 theme several 143:149 arg1 proteins 194:201 several low molecular weight transmembrane adaptor proteins 143:201 several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs 143:332 T lymphocytes express several low molecular weight transmembrane adaptor proteins that recruit src homology (SH)2 domain-containing intracellular molecules to the cell membrane via tyrosine-based signaling motifs. 10209036 4 69 theme tyrosine 535:542 arg1 phosphorylation 544:558 tyrosine phosphorylation 535:558 tyrosine phosphorylation by src and possibly syk protein tyrosine kinases 535:607 After tyrosine phosphorylation by src and possibly syk protein tyrosine kinases SIT recruits the SH2 domain-containing tyrosine phosphatase SHP2 via an immunoreceptor tyrosine-based inhibition motif. 29726814 4 0 theme subfamily 643:651 arg1 channels 653:660 other TRP subfamily channels 633:660 other TRP subfamily channels 633:660 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 2 1 theme lipid-gated 338:348 arg1 TRPC3 329:333 the full-length human TRPC3 307:333 the full-length human TRPC3 307:333 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 1 theme lipid-gated 338:348 arg1 member 355:360 a lipid-gated TRPC member 336:360 a lipid-gated TRPC member 336:360 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 4 2 from channels 653:660 arg1 mechanism 618:626 a different gating mechanism 599:626 a different gating mechanism from other TRP subfamily channels 599:660 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 5 3 theme unique 726:731 arg1 domain 747:752 a unique transmembrane domain 724:752 a unique transmembrane domain 724:752 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 6 4 theme lipid-binding 862:874 arg1 sites 876:880 two lipid-binding sites 858:880 two lipid-binding sites 858:880 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 3 5 theme first 480:484 arg1 helix 500:504 the first transmembrane helix 476:504 the first transmembrane helix 476:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 6 6 theme family 1047:1052 arg1 motif 1029:1033 the conserved LWF motif 1011:1033 the conserved LWF motif of the TRPC family 1011:1052 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 6 6 theme family 1047:1052 arg1 located 1057:1063 located 1057:1063 located 1057:1063 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 3 7 theme transmembrane 486:498 arg1 helix 500:504 the first transmembrane helix 476:504 the first transmembrane helix 476:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 6 8 theme S4-S5 937:941 arg1 linker 943:948 the S4-S5 linker 933:948 the S4-S5 linker 933:948 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 6 9 theme LWF 1025:1027 arg1 motif 1029:1033 the conserved LWF motif 1011:1033 the conserved LWF motif of the TRPC family 1011:1052 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 6 9 theme LWF 1025:1027 arg1 located 1057:1063 located 1057:1063 located 1057:1063 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 1 10 theme spinocerebellar 257:271 arg1 ataxia 273:278 spinocerebellar ataxia 257:278 spinocerebellar ataxia 257:278 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 3 11 theme prior 467:471 arg1 helices 459:465 four elbow-like membrane reentrant helices 424:465 four elbow-like membrane reentrant helices prior to the first transmembrane helix 424:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 1 12 theme TRPC 61:64 arg1 channels 66:73 The TRPC channels 57:73 The TRPC channels 57:73 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 2 13 theme lipid-occupied 368:381 arg1 state 391:395 a lipid-occupied, closed state 366:395 a lipid-occupied, closed state at 3.3 Angstrom 366:411 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 5 14 theme transmembrane 733:745 arg1 domain 747:752 a unique transmembrane domain 724:752 a unique transmembrane domain 724:752 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 4 15 theme gating 611:616 arg1 mechanism 618:626 a different gating mechanism 599:626 a different gating mechanism from other TRP subfamily channels 599:660 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 4 16 theme other 633:637 arg1 channels 653:660 other TRP subfamily channels 633:660 other TRP subfamily channels 633:660 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 5 17 theme external 826:833 arg1 stimuli 835:841 external stimuli 826:841 external stimuli 826:841 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 0 18 theme human 17:21 arg1 TRPC3 50:54 the human lipid-gated cation channel TRPC3 13:54 the human lipid-gated cation channel TRPC3 13:54 Structure of the human lipid-gated cation channel TRPC3. 29726814 1 19 theme human 182:186 arg1 diseases 188:195 human diseases 182:195 human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia 182:278 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 1 19 theme human 182:186 arg1 ataxia 273:278 spinocerebellar ataxia 257:278 spinocerebellar ataxia 257:278 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 1 19 theme human 182:186 arg1 disease 223:229 neurodegenerative disease 205:229 neurodegenerative disease 205:229 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 1 19 theme human 182:186 arg1 hypertrophy 240:250 cardiac hypertrophy 232:250 cardiac hypertrophy 232:250 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 6 20 theme pre-S1 916:921 arg1 elbow 923:927 the pre-S1 elbow 912:927 the pre-S1 elbow 912:927 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 5 21 theme third 667:671 arg1 long 710:713 long 710:713 long 710:713 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 5 21 theme third 667:671 arg1 helix 687:691 The third transmembrane helix 663:691 The third transmembrane helix S3 663:694 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 6 22 theme conserved 1015:1023 arg1 motif 1029:1033 the conserved LWF motif 1011:1033 the conserved LWF motif of the TRPC family 1011:1052 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 6 22 theme conserved 1015:1023 arg1 located 1057:1063 located 1057:1063 located 1057:1063 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 3 23 theme membrane 440:447 arg1 helices 459:465 four elbow-like membrane reentrant helices 424:465 four elbow-like membrane reentrant helices prior to the first transmembrane helix 424:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 0 24 theme cation 35:40 arg1 TRPC3 50:54 the human lipid-gated cation channel TRPC3 13:54 the human lipid-gated cation channel TRPC3 13:54 Structure of the human lipid-gated cation channel TRPC3. 29726814 2 25 theme TRPC3 329:333 arg1 structure 294:302 a structure 292:302 a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom 292:411 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 5 26 theme transmembrane 673:685 arg1 long 710:713 long 710:713 long 710:713 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 5 26 theme transmembrane 673:685 arg1 helix 687:691 The third transmembrane helix 663:691 The third transmembrane helix S3 663:694 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 3 27 contain has 420:422 arg1 TRPC3 414:418 TRPC3 414:418 TRPC3 414:418 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 3 27 contain has 420:422 arg2 helices 459:465 four elbow-like membrane reentrant helices 424:465 four elbow-like membrane reentrant helices prior to the first transmembrane helix 424:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 4 28 theme different 601:609 arg1 mechanism 618:626 a different gating mechanism 599:626 a different gating mechanism from other TRP subfamily channels 599:660 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 0 29 theme lipid-gated 23:33 arg1 TRPC3 50:54 the human lipid-gated cation channel TRPC3 13:54 the human lipid-gated cation channel TRPC3 13:54 Structure of the human lipid-gated cation channel TRPC3. 29726814 5 30 theme extracellular 775:787 arg1 sensor 816:821 a sensor 814:821 a sensor of external stimuli 814:841 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 5 30 theme extracellular 775:787 arg1 domain 789:794 an extracellular domain 772:794 an extracellular domain that may serve as a sensor of external stimuli 772:841 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 0 31 theme TRPC3 50:54 arg1 Structure 0:8 Structure 0:8 Structure of the human lipid-gated cation channel TRPC3 0:54 Structure of the human lipid-gated cation channel TRPC3. 29726814 5 32 theme stimuli 835:841 arg1 sensor 816:821 a sensor 814:821 a sensor of external stimuli 814:841 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 5 32 theme stimuli 835:841 arg1 domain 789:794 an extracellular domain 772:794 an extracellular domain that may serve as a sensor of external stimuli 772:841 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 6 33 theme TRPC 1042:1045 arg1 family 1047:1052 the TRPC family 1038:1052 the TRPC family 1038:1052 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 1 34 theme store-operated 101:114 arg1 entry 124:128 store-operated calcium entry 101:128 store-operated calcium entry 101:128 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 0 35 theme channel 42:48 arg1 TRPC3 50:54 the human lipid-gated cation channel TRPC3 13:54 the human lipid-gated cation channel TRPC3 13:54 Structure of the human lipid-gated cation channel TRPC3. 29726814 1 36 theme calcium 116:122 arg1 entry 124:128 store-operated calcium entry 101:128 store-operated calcium entry 101:128 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 4 37 theme TRP 511:513 arg1 helix 515:519 The TRP helix 507:519 The TRP helix 507:519 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 4 38 theme TRP 639:641 arg1 channels 653:660 other TRP subfamily channels 633:660 other TRP subfamily channels 633:660 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 2 39 theme human 323:327 arg1 TRPC3 329:333 the full-length human TRPC3 307:333 the full-length human TRPC3 307:333 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 39 theme human 323:327 arg1 member 355:360 a lipid-gated TRPC member 336:360 a lipid-gated TRPC member 336:360 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 1 40 theme neurodegenerative 205:221 arg1 disease 223:229 neurodegenerative disease 205:229 neurodegenerative disease 205:229 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 6 41 theme ion-conducting 984:997 arg1 other 959:963 other 959:963 other 959:963 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 6 41 theme ion-conducting 984:997 arg1 pore 999:1002 the ion-conducting pore 980:1002 the ion-conducting pore 980:1002 We identified two lipid-binding sites, one being sandwiched between the pre-S1 elbow and the S4-S5 linker, and the other being close to the ion-conducting pore, where the conserved LWF motif of the TRPC family is located. 29726814 2 42 theme full-length 311:321 arg1 TRPC3 329:333 the full-length human TRPC3 307:333 the full-length human TRPC3 307:333 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 42 theme full-length 311:321 arg1 member 355:360 a lipid-gated TRPC member 336:360 a lipid-gated TRPC member 336:360 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 3 43 theme reentrant 449:457 arg1 helices 459:465 four elbow-like membrane reentrant helices 424:465 four elbow-like membrane reentrant helices prior to the first transmembrane helix 424:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 2 44 dep lipid-occupied 368:381 arg1 closed 384:389 closed 384:389 closed 384:389 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 3 45 theme elbow-like 429:438 arg1 helices 459:465 four elbow-like membrane reentrant helices 424:465 four elbow-like membrane reentrant helices prior to the first transmembrane helix 424:504 TRPC3 has four elbow-like membrane reentrant helices prior to the first transmembrane helix. 29726814 1 46 theme calcium 134:140 arg1 homeostasis 142:152 calcium homeostasis 134:152 calcium homeostasis 134:152 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 29726814 5 47 dep helix 687:691 arg1 S3 693:694 S3 693:694 The third transmembrane helix S3 663:694 The third transmembrane helix S3 is remarkably long, shaping a unique transmembrane domain, and constituting an extracellular domain that may serve as a sensor of external stimuli. 29726814 2 48 from Angstrom 404:411 arg1 structure 294:302 a structure 292:302 a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom 292:411 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 48 from Angstrom 404:411 arg1 state 391:395 a lipid-occupied, closed state 366:395 a lipid-occupied, closed state at 3.3 Angstrom 366:411 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 4 49 theme pore-lining 572:582 arg1 S6 584:585 the pore-lining S6 568:585 the pore-lining S6 568:585 The TRP helix is perpendicular to, and thus disengaged from, the pore-lining S6, suggesting a different gating mechanism from other TRP subfamily channels. 29726814 2 50 theme TRPC 350:353 arg1 TRPC3 329:333 the full-length human TRPC3 307:333 the full-length human TRPC3 307:333 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 50 theme TRPC 350:353 arg1 member 355:360 a lipid-gated TRPC member 336:360 a lipid-gated TRPC member 336:360 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 51 from structure 294:302 arg1 state 391:395 a lipid-occupied, closed state 366:395 a lipid-occupied, closed state at 3.3 Angstrom 366:411 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 2 51 from structure 294:302 arg1 Angstrom 404:411 3.3 Angstrom 400:411 3.3 Angstrom 400:411 We present a structure of the full-length human TRPC3, a lipid-gated TRPC member, in a lipid-occupied, closed state at 3.3 Angstrom. 29726814 1 52 theme cardiac 232:238 arg1 hypertrophy 240:250 cardiac hypertrophy 232:250 cardiac hypertrophy 232:250 The TRPC channels are crucially involved in store-operated calcium entry and calcium homeostasis, and they are implicated in human diseases such as neurodegenerative disease, cardiac hypertrophy, and spinocerebellar ataxia. 23142347 2 0 theme receptors 346:354 arg1 activation 319:328 activation 319:328 activation of P1 adenosine receptors 319:354 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 1 1 theme control 241:247 arg1 point 249:253 the major control point 231:253 the major control point for extracellular adenosine levels 231:288 In vertebrates ecto-5'-nucleotidase (e5NT) catalyzes the hydrolysis of extracellular AMP to adenosine and represents the major control point for extracellular adenosine levels. 23142347 6 2 theme nucleotide-derived 893:910 arg1 compounds 932:940 nucleotide-derived and flavonoid-based compounds 893:940 nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form 893:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 2 3 theme adenosine 336:344 arg1 receptors 346:354 P1 adenosine receptors 333:354 P1 adenosine receptors 333:354 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 3 4 theme e5NT 508:511 arg1 structures 476:485 Crystal structures 468:485 Crystal structures of the dimeric human e5NT 468:511 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 4 5 theme up 707:708 arg1 motions 696:702 interchain motions 685:702 interchain motions of up to 13° 685:715 The dimerization interface is formed by the C-terminal domains and exhibits interchain motions of up to 13°. 23142347 3 6 theme Crystal 468:474 arg1 structures 476:485 Crystal structures 468:485 Crystal structures of the dimeric human e5NT 468:511 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 1 7 theme extracellular 259:271 arg1 levels 283:288 extracellular adenosine levels 259:288 extracellular adenosine levels 259:288 In vertebrates ecto-5'-nucleotidase (e5NT) catalyzes the hydrolysis of extracellular AMP to adenosine and represents the major control point for extracellular adenosine levels. 23142347 0 8 theme purinergic 92:101 arg1 signaling 103:111 purinergic signaling 92:111 purinergic signaling 92:111 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 6 9 theme Binding 876:882 arg1 modes 884:888 Binding modes 876:888 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form 876:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 5 10 theme domain 802:807 arg1 movement 809:816 the domain movement 798:816 the domain movement 798:816 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 3 11 theme dimeric 494:500 arg1 e5NT 508:511 the dimeric human e5NT 490:511 the dimeric human e5NT 490:511 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 6 12 link nucleotide-derived 893:910 arg1 compounds 932:940 nucleotide-derived and flavonoid-based compounds 893:940 nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form 893:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 6 13 theme open 984:987 arg1 form 989:992 the open form 980:992 the open form 980:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 2 14 theme hypoxia 447:453 arg1 treatment 406:414 treatment 406:414 treatment of inflammation, chronic pain, hypoxia, and cancer 406:465 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 0 15 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 0:112 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 0 15 theme Crystal 0:6 arg1 CD73 53:56 CD73 53:56 CD73 53:56 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 4 16 theme dimerization 613:624 arg1 interface 626:634 The dimerization interface 609:634 The dimerization interface 609:634 The dimerization interface is formed by the C-terminal domains and exhibits interchain motions of up to 13°. 23142347 3 17 theme 114° 533:536 arg1 switch 553:558 an extensive 114° conformational switch 520:558 an extensive 114° conformational switch between the open and closed forms of the enzyme 520:606 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 2 18 theme inflammation 419:430 arg1 treatment 406:414 treatment 406:414 treatment of inflammation, chronic pain, hypoxia, and cancer 406:465 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 1 19 theme adenosine 273:281 arg1 levels 283:288 extracellular adenosine levels 259:288 extracellular adenosine levels 259:288 In vertebrates ecto-5'-nucleotidase (e5NT) catalyzes the hydrolysis of extracellular AMP to adenosine and represents the major control point for extracellular adenosine levels. 23142347 2 20 theme pivotal 302:308 arg1 role 310:313 its pivotal role 298:313 its pivotal role for activation of P1 adenosine receptors 298:354 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 4 21 theme C-terminal 653:662 arg1 domains 664:670 the C-terminal domains 649:670 the C-terminal domains 649:670 The dimerization interface is formed by the C-terminal domains and exhibits interchain motions of up to 13°. 23142347 5 22 theme structural 776:785 arg1 control 787:793 structural control 776:793 structural control of the domain movement 776:816 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 6 23 from domain 970:975 arg1 form 989:992 the open form 980:992 the open form 980:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 6 24 theme additional 1004:1013 arg1 pocket 1023:1028 an additional binding pocket 1001:1028 an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors 1001:1097 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 1 25 theme extracellular 185:197 arg1 AMP 199:201 extracellular AMP 185:201 extracellular AMP 185:201 In vertebrates ecto-5'-nucleotidase (e5NT) catalyzes the hydrolysis of extracellular AMP to adenosine and represents the major control point for extracellular adenosine levels. 23142347 3 26 theme conformational 538:551 arg1 switch 553:558 an extensive 114° conformational switch 520:558 an extensive 114° conformational switch between the open and closed forms of the enzyme 520:606 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 0 27 theme signaling 103:111 arg1 regulation 78:87 the regulation 74:87 the regulation of purinergic signaling 74:111 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 3 28 theme extensive 523:531 arg1 switch 553:558 an extensive 114° conformational switch 520:558 an extensive 114° conformational switch between the open and closed forms of the enzyme 520:606 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 1 29 theme AMP 199:201 arg1 hydrolysis 171:180 the hydrolysis 167:180 the hydrolysis of extracellular AMP to adenosine 167:214 In vertebrates ecto-5'-nucleotidase (e5NT) catalyzes the hydrolysis of extracellular AMP to adenosine and represents the major control point for extracellular adenosine levels. 23142347 0 30 theme ecto-5'-nucleotidase 31:50 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 0:112 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 0 30 theme ecto-5'-nucleotidase 31:50 arg1 CD73 53:56 CD73 53:56 CD73 53:56 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 6 31 theme C-terminal 959:968 arg1 domain 970:975 the C-terminal domain 955:975 the C-terminal domain in the open form 955:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 5 32 theme movement 809:816 arg1 control 787:793 structural control 776:793 structural control of the domain movement 776:816 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 5 33 theme monophosphate 849:861 arg1 nucleotides 863:873 monophosphate nucleotides 849:873 monophosphate nucleotides 849:873 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 2 34 theme drug 390:393 arg1 target 395:400 an appealing drug target 377:400 an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer 377:465 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 3 35 theme open 572:575 arg1 forms 588:592 the open and closed forms 568:592 the open and closed forms of the enzyme 568:606 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 2 36 theme appealing 380:388 arg1 target 395:400 an appealing drug target 377:400 an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer 377:465 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 2 37 theme pain 441:444 arg1 treatment 406:414 treatment 406:414 treatment of inflammation, chronic pain, hypoxia, and cancer 406:465 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 3 38 theme enzyme 601:606 arg1 forms 588:592 the open and closed forms 568:592 the open and closed forms of the enzyme 568:606 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 2 39 theme P1 333:334 arg1 receptors 346:354 P1 adenosine receptors 333:354 P1 adenosine receptors 333:354 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 2 40 theme chronic 433:439 arg1 pain 441:444 chronic pain 433:444 chronic pain 433:444 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 6 41 theme compounds 932:940 arg1 modes 884:888 Binding modes 876:888 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form 876:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 5 42 with structures 726:735 arg1 AMPCP 756:760 AMPCP 756:760 AMPCP 756:760 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 5 42 with structures 726:735 arg1 adenosine 742:750 adenosine 742:750 adenosine 742:750 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 5 43 theme Complex 718:724 arg1 structures 726:735 Complex structures 718:735 Complex structures with adenosine and AMPCP 718:760 Complex structures with adenosine and AMPCP indicate that structural control of the domain movement determines the selectivity for monophosphate nucleotides. 23142347 3 44 theme human 502:506 arg1 e5NT 508:511 the dimeric human e5NT 490:511 the dimeric human e5NT 490:511 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 4 45 theme interchain 685:694 arg1 motions 696:702 interchain motions 685:702 interchain motions of up to 13° 685:715 The dimerization interface is formed by the C-terminal domains and exhibits interchain motions of up to 13°. 23142347 6 46 theme ∼210 Å 1033:1038 arg1 pocket 1023:1028 an additional binding pocket 1001:1028 an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors 1001:1097 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 0 47 dep structure 8:16 arg1 insights 60:67 insights 60:67 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 0:112 Crystal structure of the human ecto-5'-nucleotidase (CD73): insights into the regulation of purinergic signaling. 23142347 6 48 theme potent 1081:1086 arg1 inhibitors 1088:1097 more potent inhibitors 1076:1097 more potent inhibitors 1076:1097 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 6 49 theme binding 1015:1021 arg1 pocket 1023:1028 an additional binding pocket 1001:1028 an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors 1001:1097 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 2 50 theme cancer 460:465 arg1 treatment 406:414 treatment 406:414 treatment of inflammation, chronic pain, hypoxia, and cancer 406:465 Due to its pivotal role for activation of P1 adenosine receptors, e5NT has emerged as an appealing drug target for treatment of inflammation, chronic pain, hypoxia, and cancer. 23142347 6 51 theme flavonoid-based 916:930 arg1 compounds 932:940 nucleotide-derived and flavonoid-based compounds 893:940 nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form 893:992 Binding modes of nucleotide-derived and flavonoid-based compounds complexed to the C-terminal domain in the open form reveal an additional binding pocket of ∼210 Å(3) that might be explored to design more potent inhibitors. 23142347 3 52 theme closed 581:586 arg1 forms 588:592 the open and closed forms 568:592 the open and closed forms of the enzyme 568:606 Crystal structures of the dimeric human e5NT reveal an extensive 114° conformational switch between the open and closed forms of the enzyme. 23142347 1 53 theme major 235:239 arg1 point 249:253 the major control point 231:253 the major control point for extracellular adenosine levels 231:288 In vertebrates ecto-5'-nucleotidase (e5NT) catalyzes the hydrolysis of extracellular AMP to adenosine and represents the major control point for extracellular adenosine levels. 12575940 5 0 theme helix 667:671 arg1 A 673:673 helix A 667:673 helix A 667:673 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 1 theme A 673:673 arg1 shortening 653:662 a two-turn N-terminal shortening 631:662 a two-turn N-terminal shortening 631:662 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 1 theme A 673:673 arg1 feature 603:609 A striking feature 592:609 A striking feature of the structure 592:626 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 2 theme structure 618:626 arg1 feature 603:609 A striking feature 592:609 A striking feature of the structure 592:626 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 2 theme structure 618:626 arg1 shortening 653:662 a two-turn N-terminal shortening 631:662 a two-turn N-terminal shortening 631:662 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 0 3 theme heparin 84:90 arg1 activation 92:101 heparin activation 84:101 heparin activation 84:101 Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. 12575940 5 4 theme docking 722:728 arg1 studies 730:736 docking studies 722:736 docking studies 722:736 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 5 theme two-turn 633:640 arg1 shortening 653:662 a two-turn N-terminal shortening 631:662 a two-turn N-terminal shortening 631:662 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 5 theme two-turn 633:640 arg1 feature 603:609 A striking feature 592:609 A striking feature of the structure 592:626 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 0 6 theme inhibitor 31:39 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of protein C inhibitor 0:39 Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. 12575940 3 7 theme hydrophobic 434:444 arg1 hormones 446:453 hydrophobic hormones 434:453 hydrophobic hormones with apparent specificity for retinoids 434:493 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 3 8 theme unique 364:369 arg1 manner 357:362 a manner 355:362 a manner unique among the heparin binding serpins 355:403 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 3 9 theme PCI 317:319 arg1 activity 305:312 The activity 301:312 The activity of PCI 301:319 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 5 10 theme N-terminal 642:651 arg1 shortening 653:662 a two-turn N-terminal shortening 631:662 a two-turn N-terminal shortening 631:662 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 10 theme N-terminal 642:651 arg1 feature 603:609 A striking feature 592:609 A striking feature of the structure 592:626 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 1 11 theme serpin 149:154 arg1 family 156:161 the serpin family 145:161 the serpin family 145:161 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 0 12 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of protein C inhibitor 0:39 Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. 12575940 6 13 theme heparin 852:858 arg1 activation 860:869 heparin activation 852:869 heparin activation 852:869 On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. 12575940 2 14 theme seminal 247:253 arg1 vesicles 255:262 the seminal vesicles 243:262 the seminal vesicles 243:262 In blood it acts as a procoagulant, and, in the seminal vesicles, it is required for spermatogenesis. 12575940 1 15 theme family 156:161 arg1 inhibitor 114:122 Protein C inhibitor 104:122 Protein C inhibitor (PCI) 104:128 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 15 theme family 156:161 arg1 member 135:140 a member 133:140 a member of the serpin family that has many biological functions 133:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 0 16 theme protein 21:27 arg1 inhibitor 31:39 protein C inhibitor 21:39 protein C inhibitor 21:39 Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. 12575940 5 17 theme large 692:696 arg1 site 774:777 the retinoid binding site 753:777 the retinoid binding site 753:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 17 theme large 692:696 arg1 pocket 710:715 a large hydrophobic pocket 690:715 a large hydrophobic pocket that docking studies indicate to be the retinoid binding site 690:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 18 theme striking 594:601 arg1 feature 603:609 A striking feature 592:609 A striking feature of the structure 592:626 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 18 theme striking 594:601 arg1 shortening 653:662 a two-turn N-terminal shortening 631:662 a two-turn N-terminal shortening 631:662 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 3 19 theme apparent 460:467 arg1 specificity 469:479 apparent specificity 460:479 apparent specificity for retinoids 460:493 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 3 20 theme heparin 381:387 arg1 serpins 397:403 the heparin binding serpins 377:403 the heparin binding serpins 377:403 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 6 21 dep properties 818:827 arg1 basis 787:791 basis 787:791 basis 787:791 On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. 12575940 6 21 dep properties 818:827 arg1 the 783:785 the 783:785 the 783:785 On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. 12575940 1 22 theme Protein 104:110 arg1 PCI 125:127 PCI 125:127 PCI 125:127 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 22 theme Protein 104:110 arg1 inhibitor 114:122 Protein C inhibitor 104:122 Protein C inhibitor (PCI) 104:128 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 22 theme Protein 104:110 arg1 member 135:140 a member 133:140 a member of the serpin family that has many biological functions 133:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 3 23 theme binding 389:395 arg1 serpins 397:403 the heparin binding serpins 377:403 the heparin binding serpins 377:403 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 3 24 theme heparin 336:342 arg1 binding 344:350 heparin binding 336:350 heparin binding 336:350 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 6 25 theme novel 832:836 arg1 mechanism 838:846 a novel mechanism 830:846 a novel mechanism for heparin activation 830:869 On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. 12575940 4 26 theme center 561:566 arg1 RCL 574:576 RCL 574:576 RCL 574:576 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 4 26 theme center 561:566 arg1 loop 568:571 reactive center loop 552:571 reactive center loop (RCL) cleaved PCI 552:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 5 27 theme retinoid 757:764 arg1 site 774:777 the retinoid binding site 753:777 the retinoid binding site 753:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 27 theme retinoid 757:764 arg1 pocket 710:715 a large hydrophobic pocket 690:715 a large hydrophobic pocket that docking studies indicate to be the retinoid binding site 690:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 1 28 theme many 172:175 arg1 functions 188:196 many biological functions 172:196 many biological functions 172:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 0 29 theme C 29:29 arg1 inhibitor 31:39 protein C inhibitor 21:39 protein C inhibitor 21:39 Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. 12575940 5 30 theme hydrophobic 698:708 arg1 site 774:777 the retinoid binding site 753:777 the retinoid binding site 753:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 30 theme hydrophobic 698:708 arg1 pocket 710:715 a large hydrophobic pocket 690:715 a large hydrophobic pocket that docking studies indicate to be the retinoid binding site 690:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 1 31 theme biological 177:186 arg1 functions 188:196 many biological functions 172:196 many biological functions 172:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 3 32 with hormones 446:453 arg1 specificity 469:479 apparent specificity 460:479 apparent specificity for retinoids 460:493 The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. 12575940 5 33 theme binding 766:772 arg1 site 774:777 the retinoid binding site 753:777 the retinoid binding site 753:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 5 33 theme binding 766:772 arg1 pocket 710:715 a large hydrophobic pocket 690:715 a large hydrophobic pocket that docking studies indicate to be the retinoid binding site 690:777 A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. 12575940 1 34 theme C 112:112 arg1 PCI 125:127 PCI 125:127 PCI 125:127 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 34 theme C 112:112 arg1 inhibitor 114:122 Protein C inhibitor 104:122 Protein C inhibitor (PCI) 104:128 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 34 theme C 112:112 arg1 member 135:140 a member 133:140 a member of the serpin family that has many biological functions 133:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 6 35 theme electrostatic 804:816 arg1 properties 818:827 surface electrostatic properties 796:827 surface electrostatic properties 796:827 On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. 12575940 1 36 contain has 168:170 arg2 functions 188:196 many biological functions 172:196 many biological functions 172:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 36 contain has 168:170 arg1 inhibitor 114:122 Protein C inhibitor 104:122 Protein C inhibitor (PCI) 104:128 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 1 36 contain has 168:170 arg1 member 135:140 a member 133:140 a member of the serpin family that has many biological functions 133:196 Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. 12575940 0 37 theme hormone 64:70 arg1 binding 72:78 hormone binding 64:78 hormone binding 64:78 Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. 12575940 4 38 theme reactive 552:559 arg1 RCL 574:576 RCL 574:576 RCL 574:576 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 4 38 theme reactive 552:559 arg1 loop 568:571 reactive center loop 552:571 reactive center loop (RCL) cleaved PCI 552:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 4 39 theme PCI 587:589 arg1 structure 539:547 the 2.4 A crystallographic structure 512:547 the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI 512:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 4 40 theme loop 568:571 arg1 PCI 587:589 reactive center loop (RCL) cleaved PCI 552:589 reactive center loop (RCL) cleaved PCI 552:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 4 41 theme crystallographic 522:537 arg1 structure 539:547 the 2.4 A crystallographic structure 512:547 the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI 512:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 6 42 theme surface 796:802 arg1 properties 818:827 surface electrostatic properties 796:827 surface electrostatic properties 796:827 On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. 12575940 4 43 theme cleaved 579:585 arg1 PCI 587:589 reactive center loop (RCL) cleaved PCI 552:589 reactive center loop (RCL) cleaved PCI 552:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 12575940 4 44 theme A 520:520 arg1 structure 539:547 the 2.4 A crystallographic structure 512:547 the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI 512:589 Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. 19299620 7 0 theme antibody-based 1126:1139 arg1 therapy 1141:1147 antibody-based therapy 1126:1147 antibody-based therapy 1126:1147 They could also provide new opportunities for antibody-based therapy. 19299620 4 1 theme Crystallographic 596:611 arg1 studies 629:635 Crystallographic and mutagenesis studies 596:635 Crystallographic and mutagenesis studies 596:635 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 4 2 theme mutagenesis 617:627 arg1 studies 629:635 Crystallographic and mutagenesis studies 596:635 Crystallographic and mutagenesis studies 596:635 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 4 3 dep areas 798:802 arg1 overlap 761:767 overlap 761:767 overlap 761:767 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 2 4 theme binding 293:299 arg1 site 301:304 an antigen binding site 282:304 an antigen binding site that binds two distinct proteins with high affinity 282:356 Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. 19299620 0 5 theme antigen 75:81 arg1 site 91:94 the antigen binding site 71:94 the antigen binding site 71:94 Variants of the antibody herceptin that interact with HER2 and VEGF at the antigen binding site. 19299620 2 6 with proteins 330:337 arg1 affinity 349:356 high affinity 344:356 high affinity 344:356 Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. 19299620 4 7 theme surface 790:796 arg1 areas 798:802 the antibody surface areas 777:802 the antibody surface areas contacting the two antigens 777:830 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 6 8 theme site 1061:1064 arg1 paradigm 1037:1044 the monoclonal antibody paradigm 1013:1044 the monoclonal antibody paradigm of one binding site, one antigen 1013:1077 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 7 9 theme new 1104:1106 arg1 opportunities 1108:1120 new opportunities 1104:1120 new opportunities for antibody-based therapy 1104:1147 They could also provide new opportunities for antibody-based therapy. 19299620 4 10 theme antibody 781:788 arg1 areas 798:802 the antibody surface areas 777:802 the antibody surface areas contacting the two antigens 777:830 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 4 11 theme antibody 680:687 arg1 acids 666:670 distinct amino acids 651:670 distinct amino acids 651:670 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 6 12 theme binding 1053:1059 arg1 site 1061:1064 one binding site 1049:1064 one binding site 1049:1064 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 6 12 theme binding 1053:1059 arg1 antigen 1071:1077 one antigen 1067:1077 one antigen 1067:1077 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 3 13 theme factor 467:472 arg1 receptor 474:481 the human epidermal growth factor receptor 2 440:483 the human epidermal growth factor receptor 2 (HER2) 440:490 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 13 theme factor 467:472 arg1 HER2 486:489 HER2 486:489 HER2 486:489 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 6 14 theme two-in-one 980:989 arg1 antibodies 992:1001 Such "two-in-one" antibodies 974:1001 Such "two-in-one" antibodies 974:1001 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 0 15 theme binding 83:89 arg1 site 91:94 the antigen binding site 71:94 the antigen binding site 71:94 Variants of the antibody herceptin that interact with HER2 and VEGF at the antigen binding site. 19299620 5 16 theme VEGF-mediated 893:905 arg1 proliferation 912:924 VEGF-mediated cell proliferation 893:924 VEGF-mediated cell proliferation 893:924 An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. 19299620 6 17 theme " 990:990 arg1 antibodies 992:1001 Such "two-in-one" antibodies 974:1001 Such "two-in-one" antibodies 974:1001 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 2 18 theme distinct 321:328 arg1 proteins 330:337 two distinct proteins 317:337 two distinct proteins with high affinity 317:356 Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. 19299620 4 19 theme amino 660:664 arg1 acids 666:670 distinct amino acids 651:670 distinct amino acids 651:670 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 3 20 theme growth 574:579 arg1 VEGF 589:592 VEGF 589:592 VEGF 589:592 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 20 theme growth 574:579 arg1 factor 581:586 vascular endothelial growth factor 553:586 vascular endothelial growth factor (VEGF) 553:593 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 0 21 theme antibody 16:23 arg1 herceptin 25:33 the antibody herceptin 12:33 the antibody herceptin 12:33 Variants of the antibody herceptin that interact with HER2 and VEGF at the antigen binding site. 19299620 5 22 theme bH1 865:867 arg1 version 854:860 An affinity-improved version 833:860 An affinity-improved version of bH1 833:867 An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. 19299620 3 23 dep ability 513:519 arg1 the 496:498 the 496:498 the 496:498 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 23 dep ability 513:519 arg1 basis 500:504 basis 500:504 basis 500:504 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 24 theme endothelial 562:572 arg1 VEGF 589:592 VEGF 589:592 VEGF 589:592 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 24 theme endothelial 562:572 arg1 factor 581:586 vascular endothelial growth factor 553:586 vascular endothelial growth factor (VEGF) 553:593 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 25 theme human 444:448 arg1 receptor 474:481 the human epidermal growth factor receptor 2 440:483 the human epidermal growth factor receptor 2 (HER2) 440:490 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 25 theme human 444:448 arg1 HER2 486:489 HER2 486:489 HER2 486:489 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 26 theme therapeutic 397:407 arg1 antibody 420:427 a therapeutic monoclonal antibody 395:427 a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2) 395:490 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 26 theme therapeutic 397:407 arg1 Herceptin 384:392 Herceptin 384:392 Herceptin 384:392 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 2 27 with antibody 268:275 arg1 site 301:304 an antigen binding site 282:304 an antigen binding site that binds two distinct proteins with high affinity 282:356 Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. 19299620 3 28 theme Herceptin 384:392 arg1 variant 373:379 a variant 371:379 a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), 371:491 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 0 29 theme herceptin 25:33 arg1 Variants 0:7 Variants 0:7 Variants of the antibody herceptin that interact with HER2 and VEGF at the antigen binding site 0:94 Variants of the antibody herceptin that interact with HER2 and VEGF at the antigen binding site. 19299620 5 30 theme tumor 939:943 arg1 progression 945:955 tumor progression 939:955 tumor progression 939:955 An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. 19299620 2 31 theme high 344:347 arg1 affinity 349:356 high affinity 344:356 high affinity 344:356 Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. 19299620 3 32 theme vascular 553:560 arg1 VEGF 589:592 VEGF 589:592 VEGF 589:592 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 32 theme vascular 553:560 arg1 factor 581:586 vascular endothelial growth factor 553:586 vascular endothelial growth factor (VEGF) 553:593 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 6 33 theme antibody 1028:1035 arg1 paradigm 1037:1044 the monoclonal antibody paradigm 1013:1044 the monoclonal antibody paradigm of one binding site, one antigen 1013:1077 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 2 34 theme antigen 285:291 arg1 site 301:304 an antigen binding site 282:304 an antigen binding site that binds two distinct proteins with high affinity 282:356 Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. 19299620 5 35 theme mouse 960:964 arg1 models 966:971 mouse models 960:971 mouse models 960:971 An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. 19299620 5 36 theme cell 907:910 arg1 proliferation 912:924 VEGF-mediated cell proliferation 893:924 VEGF-mediated cell proliferation 893:924 An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. 19299620 3 37 theme monoclonal 409:418 arg1 antibody 420:427 a therapeutic monoclonal antibody 395:427 a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2) 395:490 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 37 theme monoclonal 409:418 arg1 Herceptin 384:392 Herceptin 384:392 Herceptin 384:392 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 5 38 theme affinity-improved 836:852 arg1 version 854:860 An affinity-improved version 833:860 An affinity-improved version of bH1 833:867 An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. 19299620 4 39 theme distinct 651:658 arg1 acids 666:670 distinct amino acids 651:670 distinct amino acids 651:670 Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. 19299620 6 40 theme monoclonal 1017:1026 arg1 antibody 1028:1035 the monoclonal antibody 1013:1035 the monoclonal antibody paradigm of one binding site, one antigen 1013:1077 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 6 41 theme Such 974:977 arg1 antibodies 992:1001 Such "two-in-one" antibodies 974:1001 Such "two-in-one" antibodies 974:1001 Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. 19299620 3 42 theme epidermal 450:458 arg1 receptor 474:481 the human epidermal growth factor receptor 2 440:483 the human epidermal growth factor receptor 2 (HER2) 440:490 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 42 theme epidermal 450:458 arg1 HER2 486:489 HER2 486:489 HER2 486:489 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 43 theme growth 460:465 arg1 receptor 474:481 the human epidermal growth factor receptor 2 440:483 the human epidermal growth factor receptor 2 (HER2) 440:490 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 3 43 theme growth 460:465 arg1 HER2 486:489 HER2 486:489 HER2 486:489 We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). 19299620 1 44 theme antibody 196:203 arg1 surface 205:211 an antibody surface 193:211 an antibody surface 193:211 The interface between antibody and antigen is often depicted as a lock and key, suggesting that an antibody surface can accommodate only one antigen. 15093539 0 0 theme complex 72:78 arg1 structure 38:46 the structure 34:46 the structure of the ErbB2-pertuzumab complex 34:78 Insights into ErbB signaling from the structure of the ErbB2-pertuzumab complex. 15093539 1 1 theme anti-ErbB2 291:300 arg1 antibody 313:320 an anti-ErbB2 monoclonal antibody 288:320 an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 288:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 1 theme anti-ErbB2 291:300 arg1 pertuzumab 276:285 pertuzumab 276:285 pertuzumab 276:285 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 4 2 theme ErbB2-ErbB3 807:817 arg1 heterodimerization 819:836 ErbB2-ErbB3 heterodimerization 807:836 ErbB2-ErbB3 heterodimerization 807:836 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 1 3 theme extracellular 141:153 arg1 domain 155:160 the extracellular domain 137:160 the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 137:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 3 theme extracellular 141:153 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 4 theme monoclonal 302:311 arg1 antibody 313:320 an anti-ErbB2 monoclonal antibody 288:320 an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 288:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 4 theme monoclonal 302:311 arg1 pertuzumab 276:285 pertuzumab 276:285 pertuzumab 276:285 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 4 5 theme residues 706:713 arg1 number 696:701 a number 694:701 a number of residues contacting pertuzumab 694:735 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 5 6 theme receptor 923:930 arg1 homodimerization 932:947 EGF receptor homodimerization 919:947 EGF receptor homodimerization 919:947 We have also shown that conserved residues previously shown to be necessary for EGF receptor homodimerization may be dispensible for ErbB2-ErbB3 heterodimerization. 15093539 2 7 theme receptor 460:467 arg1 dimerization 469:480 receptor dimerization 460:480 receptor dimerization 460:480 Pertuzumab binds to ErbB2 near the center of domain II, sterically blocking a binding pocket necessary for receptor dimerization and signaling. 15093539 0 8 from structure 38:46 arg1 signaling 19:27 ErbB signaling 14:27 ErbB signaling from the structure of the ErbB2-pertuzumab complex 14:78 Insights into ErbB signaling from the structure of the ErbB2-pertuzumab complex. 15093539 1 9 theme domain 155:160 arg1 structure 124:132 A X-ray crystal structure 108:132 the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 100:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 2 10 theme necessary 446:454 arg1 pocket 439:444 a binding pocket 429:444 a binding pocket necessary for receptor dimerization and signaling 429:494 Pertuzumab binds to ErbB2 near the center of domain II, sterically blocking a binding pocket necessary for receptor dimerization and signaling. 15093539 5 11 theme conserved 863:871 arg1 residues 873:880 conserved residues 863:880 conserved residues previously shown to be necessary for EGF receptor homodimerization 863:947 We have also shown that conserved residues previously shown to be necessary for EGF receptor homodimerization may be dispensible for ErbB2-ErbB3 heterodimerization. 15093539 4 12 theme pertuzumab 784:793 arg1 binding 795:801 pertuzumab binding 784:801 pertuzumab binding 784:801 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 1 13 from complex 231:237 arg1 domain 155:160 the extracellular domain 137:160 the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 137:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 13 from complex 231:237 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 14 with complex 231:237 arg1 fragment 264:271 the antigen binding fragment 244:271 the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 244:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 15 theme human 169:173 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 4 16 theme mutations 771:779 arg1 effects 754:760 the effects 750:760 the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization 750:836 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 3 17 theme ErbB2-pertuzumab 501:516 arg1 structure 518:526 The ErbB2-pertuzumab structure 497:526 The ErbB2-pertuzumab structure 497:526 The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. 15093539 1 18 theme epidermal 175:183 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 0 19 theme ErbB 14:17 arg1 signaling 19:27 ErbB signaling 14:27 ErbB signaling from the structure of the ErbB2-pertuzumab complex 14:78 Insights into ErbB signaling from the structure of the ErbB2-pertuzumab complex. 15093539 5 20 theme EGF 919:921 arg1 homodimerization 932:947 EGF receptor homodimerization 919:947 EGF receptor homodimerization 919:947 We have also shown that conserved residues previously shown to be necessary for EGF receptor homodimerization may be dispensible for ErbB2-ErbB3 heterodimerization. 15093539 1 21 theme growth 185:190 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 22 theme antigen 248:254 arg1 fragment 264:271 the antigen binding fragment 244:271 the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 244:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 3 23 theme ErbB2 615:619 arg1 interaction 621:631 ErbB2 interaction 615:631 ErbB2 interaction 615:631 The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. 15093539 3 24 theme mutagenesis 551:561 arg1 data 563:566 earlier mutagenesis data 543:566 earlier mutagenesis data 543:566 The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. 15093539 1 25 theme factor 192:197 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 26 theme binding 256:262 arg1 fragment 264:271 the antigen binding fragment 244:271 the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 244:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 3 27 theme pertuzumab 581:590 arg1 residues 592:599 the pertuzumab residues 577:599 the pertuzumab residues essential for ErbB2 interaction 577:631 The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. 15093539 1 28 theme receptor 199:206 arg1 domain 155:160 the extracellular domain 137:160 the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 137:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 28 theme receptor 199:206 arg1 receptor 199:206 the human epidermal growth factor receptor 2 165:208 the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 165:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 3 29 theme essential 601:609 arg1 residues 592:599 the pertuzumab residues 577:599 the pertuzumab residues essential for ErbB2 interaction 577:631 The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. 15093539 1 30 from receptor 199:206 arg1 complex 231:237 a complex 229:237 a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 229:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 4 31 theme interface 667:675 arg1 side 655:658 the ErbB2 side 645:658 the ErbB2 side of the interface 645:675 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 2 32 theme domain 398:403 arg1 center 388:393 the center 384:393 the center of domain II 384:406 Pertuzumab binds to ErbB2 near the center of domain II, sterically blocking a binding pocket necessary for receptor dimerization and signaling. 15093539 3 33 theme earlier 543:549 arg1 data 563:566 earlier mutagenesis data 543:566 earlier mutagenesis data 543:566 The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. 15093539 1 34 theme pertuzumab 276:285 arg1 fragment 264:271 the antigen binding fragment 244:271 the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 244:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 35 theme X-ray 110:114 arg1 structure 124:132 A X-ray crystal structure 108:132 the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 100:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 36 dep receptor 199:206 arg1 ErbB2 211:215 ErbB2 211:215 ErbB2 211:215 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 1 36 dep receptor 199:206 arg1 HER2 220:223 HER2 220:223 HER2 220:223 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 4 37 from effects 754:760 arg1 binding 795:801 pertuzumab binding 784:801 pertuzumab binding 784:801 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 4 37 from effects 754:760 arg1 heterodimerization 819:836 ErbB2-ErbB3 heterodimerization 807:836 ErbB2-ErbB3 heterodimerization 807:836 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 1 38 theme crystal 116:122 arg1 structure 124:132 A X-ray crystal structure 108:132 the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 100:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 15093539 0 39 theme ErbB2-pertuzumab 55:70 arg1 complex 72:78 the ErbB2-pertuzumab complex 51:78 the ErbB2-pertuzumab complex 51:78 Insights into ErbB signaling from the structure of the ErbB2-pertuzumab complex. 15093539 5 40 theme ErbB2-ErbB3 972:982 arg1 heterodimerization 984:1001 ErbB2-ErbB3 heterodimerization 972:1001 ErbB2-ErbB3 heterodimerization 972:1001 We have also shown that conserved residues previously shown to be necessary for EGF receptor homodimerization may be dispensible for ErbB2-ErbB3 heterodimerization. 15093539 4 41 theme ErbB2 649:653 arg1 side 655:658 the ErbB2 side 645:658 the ErbB2 side of the interface 645:675 To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. 15093539 2 42 theme binding 431:437 arg1 pocket 439:444 a binding pocket 429:444 a binding pocket necessary for receptor dimerization and signaling 429:494 Pertuzumab binds to ErbB2 near the center of domain II, sterically blocking a binding pocket necessary for receptor dimerization and signaling. 15093539 1 43 from domain 155:160 arg1 complex 231:237 a complex 229:237 a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg 229:350 We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. 35294879 0 0 theme enzyme 99:104 arg1 activity 106:113 its enzyme activity 95:113 its enzyme activity 95:113 Extracellular endosulfatase Sulf-2 harbors a chondroitin/dermatan sulfate chain that modulates its enzyme activity. 35294879 1 1 theme multiple 287:294 arg1 processes 315:323 multiple physiopathological processes 287:323 multiple physiopathological processes 287:323 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 1 1 theme multiple 287:294 arg1 cancer 336:341 cancer 336:341 cancer 336:341 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 5 2 theme GAG 843:845 arg1 chain 847:851 its GAG chain 839:851 its GAG chain 839:851 In addition, we show that mammalian hyaluronidase acts as a promoter of HSulf-2 activity by digesting its GAG chain. 35294879 2 3 theme major 401:405 arg1 discrepancies 407:419 major discrepancies 401:419 major discrepancies regarding their in vivo functions 401:453 However, Sulfs remain poorly characterized enzymes, with major discrepancies regarding their in vivo functions. 35294879 1 4 theme physiopathological 296:313 arg1 processes 315:323 multiple physiopathological processes 287:323 multiple physiopathological processes 287:323 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 1 4 theme physiopathological 296:313 arg1 cancer 336:341 cancer 336:341 cancer 336:341 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 6 5 theme critical 955:962 arg1 modulator 978:986 a critical non-catalytic modulator 953:986 a critical non-catalytic modulator of the enzyme activity 953:1009 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 6 5 theme critical 955:962 arg1 HSulf-2 891:897 HSulf-2 891:897 HSulf-2 891:897 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 3 6 theme substrate-binding 590:606 arg1 domain 608:613 the enzyme substrate-binding domain 579:613 the enzyme substrate-binding domain 579:613 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 4 7 theme HSulf-2 698:704 arg1 activity 706:713 HSulf-2 activity in vitro and in vivo 698:734 activity 706:713 We demonstrate that this GAG chain affects enzyme/substrate recognition and tunes HSulf-2 activity in vitro and in vivo. 35294879 3 8 theme sulfate 528:534 arg1 GAG 555:557 GAG 555:557 GAG 555:557 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 3 8 theme sulfate 528:534 arg1 glycosaminoglycan 536:552 chondroitin/dermatan sulfate glycosaminoglycan 507:552 a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain 505:564 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 1 9 theme original 186:193 arg1 mechanism 221:229 an original post-synthetic regulatory mechanism 183:229 an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides 183:265 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 5 10 theme HSulf-2 809:815 arg1 activity 817:824 HSulf-2 activity 809:824 HSulf-2 activity 809:824 In addition, we show that mammalian hyaluronidase acts as a promoter of HSulf-2 activity by digesting its GAG chain. 35294879 4 11 theme GAG 641:643 arg1 chain 645:649 this GAG chain 636:649 this GAG chain 636:649 We demonstrate that this GAG chain affects enzyme/substrate recognition and tunes HSulf-2 activity in vitro and in vivo. 35294879 3 12 theme enzyme 583:588 arg1 domain 608:613 the enzyme substrate-binding domain 579:613 the enzyme substrate-binding domain 579:613 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 1 13 theme post-synthetic 195:208 arg1 mechanism 221:229 an original post-synthetic regulatory mechanism 183:229 an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides 183:265 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 0 14 theme endosulfatase 14:26 arg1 Sulf-2 28:33 Extracellular endosulfatase Sulf-2 0:33 Extracellular endosulfatase Sulf-2 0:33 Extracellular endosulfatase Sulf-2 harbors a chondroitin/dermatan sulfate chain that modulates its enzyme activity. 35294879 6 15 theme GAG 940:942 arg1 chain 944:948 its GAG chain 936:948 its GAG chain 936:948 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 1 16 theme regulatory 210:219 arg1 mechanism 221:229 an original post-synthetic regulatory mechanism 183:229 an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides 183:265 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 0 17 theme Extracellular 0:12 arg1 Sulf-2 28:33 Extracellular endosulfatase Sulf-2 0:33 Extracellular endosulfatase Sulf-2 0:33 Extracellular endosulfatase Sulf-2 harbors a chondroitin/dermatan sulfate chain that modulates its enzyme activity. 35294879 2 18 theme in vivo 437:443 arg1 functions 445:453 their in vivo functions 431:453 their in vivo functions 431:453 However, Sulfs remain poorly characterized enzymes, with major discrepancies regarding their in vivo functions. 35294879 5 19 theme mammalian 763:771 arg1 hyaluronidase 773:785 mammalian hyaluronidase 763:785 mammalian hyaluronidase 763:785 In addition, we show that mammalian hyaluronidase acts as a promoter of HSulf-2 activity by digesting its GAG chain. 35294879 5 19 theme mammalian 763:771 arg1 promoter 797:804 a promoter 795:804 a promoter of HSulf-2 activity 795:824 In addition, we show that mammalian hyaluronidase acts as a promoter of HSulf-2 activity by digesting its GAG chain. 35294879 3 20 theme chondroitin/dermatan 507:526 arg1 GAG 555:557 GAG 555:557 GAG 555:557 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 3 20 theme chondroitin/dermatan 507:526 arg1 glycosaminoglycan 536:552 chondroitin/dermatan sulfate glycosaminoglycan 507:552 a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain 505:564 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 2 21 with enzymes 387:393 arg1 discrepancies 407:419 major discrepancies 401:419 major discrepancies regarding their in vivo functions 401:453 However, Sulfs remain poorly characterized enzymes, with major discrepancies regarding their in vivo functions. 35294879 1 22 theme heparan 235:241 arg1 polysaccharides 251:265 heparan sulfate polysaccharides 235:265 heparan sulfate polysaccharides 235:265 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 0 23 theme chondroitin/dermatan 45:64 arg1 chain 74:78 a chondroitin/dermatan sulfate chain 43:78 a chondroitin/dermatan sulfate chain that modulates its enzyme activity 43:113 Extracellular endosulfatase Sulf-2 harbors a chondroitin/dermatan sulfate chain that modulates its enzyme activity. 35294879 7 24 from data 1068:1071 arg1 activities 1080:1089 the activities 1076:1089 the activities of the Sulfs 1076:1102 These findings contribute to clarifying the conflicting data on the activities of the Sulfs. 35294879 1 25 theme sulfate 243:249 arg1 polysaccharides 251:265 heparan sulfate polysaccharides 235:265 heparan sulfate polysaccharides 235:265 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 3 26 theme human 474:478 arg1 HSulf-2 488:494 HSulf-2 488:494 HSulf-2 488:494 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 3 26 theme human 474:478 arg1 Sulf-2 480:485 human Sulf-2 474:485 human Sulf-2 (HSulf-2) 474:495 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 6 27 theme activity 1002:1009 arg1 modulator 978:986 a critical non-catalytic modulator 953:986 a critical non-catalytic modulator of the enzyme activity 953:1009 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 6 27 theme activity 1002:1009 arg1 HSulf-2 891:897 HSulf-2 891:897 HSulf-2 891:897 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 6 28 theme proteoglycan-related 904:923 arg1 enzyme 925:930 a proteoglycan-related enzyme 902:930 a proteoglycan-related enzyme 902:930 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 6 28 theme proteoglycan-related 904:923 arg1 HSulf-2 891:897 HSulf-2 891:897 HSulf-2 891:897 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 4 29 dep activity 706:713 arg1 in vitro 715:722 in vitro 715:722 in vitro 715:722 We demonstrate that this GAG chain affects enzyme/substrate recognition and tunes HSulf-2 activity in vitro and in vivo. 35294879 3 30 attach attached 567:574 arg1 domain 608:613 the enzyme substrate-binding domain 579:613 the enzyme substrate-binding domain 579:613 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 3 30 attach attached 567:574 arg2 chain 560:564 a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain 505:564 a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain 505:564 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 0 31 theme sulfate 66:72 arg1 chain 74:78 a chondroitin/dermatan sulfate chain 43:78 a chondroitin/dermatan sulfate chain that modulates its enzyme activity 43:113 Extracellular endosulfatase Sulf-2 harbors a chondroitin/dermatan sulfate chain that modulates its enzyme activity. 35294879 6 32 theme enzyme 995:1000 arg1 activity 1002:1009 the enzyme activity 991:1009 the enzyme activity 991:1009 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 6 33 theme non-catalytic 964:976 arg1 modulator 978:986 a critical non-catalytic modulator 953:986 a critical non-catalytic modulator of the enzyme activity 953:1009 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 6 33 theme non-catalytic 964:976 arg1 HSulf-2 891:897 HSulf-2 891:897 HSulf-2 891:897 In conclusion, our results highlight HSulf-2 as a proteoglycan-related enzyme and its GAG chain as a critical non-catalytic modulator of the enzyme activity. 35294879 3 34 theme glycosaminoglycan 536:552 arg1 chain 560:564 a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain 505:564 a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain 505:564 Here we show that human Sulf-2 (HSulf-2) harbors a chondroitin/dermatan sulfate glycosaminoglycan (GAG) chain, attached to the enzyme substrate-binding domain. 35294879 7 35 theme Sulfs 1098:1102 arg1 activities 1080:1089 the activities 1076:1089 the activities of the Sulfs 1076:1102 These findings contribute to clarifying the conflicting data on the activities of the Sulfs. 35294879 7 36 theme conflicting 1056:1066 arg1 data 1068:1071 the conflicting data 1052:1071 the conflicting data on the activities of the Sulfs 1052:1102 These findings contribute to clarifying the conflicting data on the activities of the Sulfs. 35294879 1 37 theme unconventional 143:156 arg1 sulfatases 158:167 unconventional sulfatases 143:167 unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer 143:341 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 35294879 5 38 theme activity 817:824 arg1 hyaluronidase 773:785 mammalian hyaluronidase 763:785 mammalian hyaluronidase 763:785 In addition, we show that mammalian hyaluronidase acts as a promoter of HSulf-2 activity by digesting its GAG chain. 35294879 5 38 theme activity 817:824 arg1 promoter 797:804 a promoter 795:804 a promoter of HSulf-2 activity 795:824 In addition, we show that mammalian hyaluronidase acts as a promoter of HSulf-2 activity by digesting its GAG chain. 35294879 1 39 theme sulfatases 158:167 arg1 class 134:138 a class 132:138 a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer 132:341 Sulfs represent a class of unconventional sulfatases which provide an original post-synthetic regulatory mechanism for heparan sulfate polysaccharides and are involved in multiple physiopathological processes, including cancer. 2013294 9 0 theme 14-kDa 840:845 arg1 protein 867:873 a 14-kDa submandibular gland protein 838:873 a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 838:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 5 1 theme N-terminal 547:556 arg1 residue 558:564 the N-terminal residue 543:564 the N-terminal residue 543:564 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 1 theme N-terminal 547:556 arg1 acids 481:485 118 amino acids 471:485 118 amino acids with a calculated Mr of 13,506 471:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 1 theme N-terminal 547:556 arg1 acid 535:538 pyroglutamic acid 522:538 pyroglutamic acid 522:538 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 1 2 theme human 136:140 arg1 plasma 150:155 human seminal plasma 136:155 human seminal plasma 136:155 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 10 3 theme disease 1008:1014 arg1 protein 1022:1028 gross cystic disease fluid protein 15 995:1031 gross cystic disease fluid protein 15 (sequences translated from cDNA clones) 995:1071 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 7 4 theme carbohydrate 634:645 arg1 composition 647:657 The carbohydrate composition 630:657 The carbohydrate composition 630:657 The carbohydrate composition shows an unusually high amount of fucose. 2013294 9 5 theme submandibular 847:859 arg1 gland 861:865 submandibular gland 847:865 a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 838:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 0 6 from structure 8:16 arg1 plasma 68:73 human seminal plasma 54:73 human seminal plasma 54:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 1 7 theme seminal 142:148 arg1 plasma 150:155 human seminal plasma 136:155 human seminal plasma 136:155 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 10 8 theme breast 1090:1095 arg1 tissues 1097:1103 human breast tissues 1084:1103 human breast tissues 1084:1103 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 9 9 theme gland 861:865 arg1 protein 867:873 a 14-kDa submandibular gland protein 838:873 a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 838:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 4 10 theme SABP 311:314 arg1 sequence 299:306 The complete amino acid sequence 275:306 The complete amino acid sequence of SABP 275:314 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 10 11 theme human 1084:1088 arg1 tissues 1097:1103 human breast tissues 1084:1103 human breast tissues 1084:1103 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 4 12 theme aureus 394:399 arg1 protease 404:411 Staphylococcus aureus V8 protease 379:411 Staphylococcus aureus V8 protease 379:411 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 5 13 theme 13,506 511:516 arg1 Mr 505:506 a calculated Mr 492:506 a calculated Mr of 13,506 492:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 4 14 theme Staphylococcus 379:392 arg1 protease 404:411 Staphylococcus aureus V8 protease 379:411 Staphylococcus aureus V8 protease 379:411 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 5 15 theme pyroglutamic 522:533 arg1 residue 558:564 the N-terminal residue 543:564 the N-terminal residue 543:564 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 15 theme pyroglutamic 522:533 arg1 acids 481:485 118 amino acids 471:485 118 amino acids with a calculated Mr of 13,506 471:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 15 theme pyroglutamic 522:533 arg1 acid 535:538 pyroglutamic acid 522:538 pyroglutamic acid 522:538 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 11 16 theme seminal 1192:1198 arg1 vesicles 1200:1207 seminal vesicles 1192:1207 seminal vesicles 1192:1207 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 6 17 from Asn77 623:627 arg1 located 612:618 located 612:618 located 612:618 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 6 17 from Asn77 623:627 arg1 moiety 602:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 11 18 located detected 1129:1136 arg1 extracts 1152:1159 extracts 1152:1159 extracts of the submandibular gland and seminal vesicles 1152:1207 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 11 18 located detected 1129:1136 arg2 SABP 1115:1118 SABP 1115:1118 SABP 1115:1118 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 11 18 located detected 1129:1136 arg1 saliva 1141:1146 saliva 1141:1146 saliva 1141:1146 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 10 19 theme gross 995:999 arg1 protein 1022:1028 gross cystic disease fluid protein 15 995:1031 gross cystic disease fluid protein 15 (sequences translated from cDNA clones) 995:1071 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 4 20 theme acid 294:297 arg1 sequence 299:306 The complete amino acid sequence 275:306 The complete amino acid sequence of SABP 275:314 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 9 21 theme 45 887:888 arg1 % 889:889 % 889:889 % 889:889 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 9 22 with similarity 822:831 arg1 protein 867:873 a 14-kDa submandibular gland protein 838:873 a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 838:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 4 23 theme V8 401:402 arg1 protease 404:411 Staphylococcus aureus V8 protease 379:411 Staphylococcus aureus V8 protease 379:411 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 4 24 theme amino 288:292 arg1 sequence 299:306 The complete amino acid sequence 275:306 The complete amino acid sequence of SABP 275:314 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 1 25 theme Secretory 76:84 arg1 SABP 109:112 SABP 109:112 SABP 109:112 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 1 25 theme Secretory 76:84 arg1 protein 100:106 Secretory actin-binding protein 76:106 Secretory actin-binding protein (SABP) 76:113 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 1 25 theme Secretory 76:84 arg1 glycoprotein 118:129 a glycoprotein 116:129 a glycoprotein from human seminal plasma 116:155 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 9 26 dep mouse 880:884 arg1 identity 891:898 45% identity 887:898 45% identity 887:898 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 9 26 dep mouse 880:884 arg1 similarity 908:917 64% similarity 904:917 64% similarity 904:917 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 10 27 theme cystic 1001:1006 arg1 protein 1022:1028 gross cystic disease fluid protein 15 995:1031 gross cystic disease fluid protein 15 (sequences translated from cDNA clones) 995:1071 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 0 28 theme Primary 0:6 arg1 structure 8:16 Primary structure 0:16 Primary structure of a new actin-binding protein from human seminal plasma 0:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 6 29 located located 612:618 arg2 located 612:618 located 612:618 located 612:618 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 6 29 located located 612:618 arg2 moiety 602:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 6 29 located located 612:618 arg1 Asn77 623:627 Asn77 623:627 Asn77 623:627 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 1 30 theme actin-binding 86:98 arg1 SABP 109:112 SABP 109:112 SABP 109:112 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 1 30 theme actin-binding 86:98 arg1 protein 100:106 Secretory actin-binding protein 76:106 Secretory actin-binding protein (SABP) 76:113 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 1 30 theme actin-binding 86:98 arg1 glycoprotein 118:129 a glycoprotein 116:129 a glycoprotein from human seminal plasma 116:155 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 9 31 from mouse 880:884 arg1 protein 867:873 a 14-kDa submandibular gland protein 838:873 a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 838:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 9 32 theme % 889:889 arg1 identity 891:898 45% identity 887:898 45% identity 887:898 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 9 33 theme Sequence 776:783 arg1 comparison 785:794 Sequence comparison 776:794 Sequence comparison 776:794 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 5 34 theme calculated 494:503 arg1 Mr 505:506 a calculated Mr 492:506 a calculated Mr of 13,506 492:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 6 35 theme carbohydrate 589:600 arg1 located 612:618 located 612:618 located 612:618 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 6 35 theme carbohydrate 589:600 arg1 moiety 602:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 11 36 theme vesicles 1200:1207 arg1 extracts 1152:1159 extracts 1152:1159 extracts of the submandibular gland and seminal vesicles 1152:1207 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 5 37 with acids 481:485 arg1 Mr 505:506 a calculated Mr 492:506 a calculated Mr of 13,506 492:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 6 38 theme N-glycosidic 576:587 arg1 located 612:618 located 612:618 located 612:618 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 6 38 theme N-glycosidic 576:587 arg1 moiety 602:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 11 39 theme gland 1182:1186 arg1 extracts 1152:1159 extracts 1152:1159 extracts of the submandibular gland and seminal vesicles 1152:1207 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 0 40 theme actin-binding 27:39 arg1 protein 41:47 a new actin-binding protein 21:47 a new actin-binding protein from human seminal plasma 21:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 10 41 theme fluid 1016:1020 arg1 protein 1022:1028 gross cystic disease fluid protein 15 995:1031 gross cystic disease fluid protein 15 (sequences translated from cDNA clones) 995:1071 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 0 42 from plasma 68:73 arg1 protein 41:47 a new actin-binding protein 21:47 a new actin-binding protein from human seminal plasma 21:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 0 42 from plasma 68:73 arg1 structure 8:16 Primary structure 0:16 Primary structure of a new actin-binding protein from human seminal plasma 0:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 6 43 theme single 569:574 arg1 located 612:618 located 612:618 located 612:618 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 6 43 theme single 569:574 arg1 moiety 602:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety 567:607 A single N-glycosidic carbohydrate moiety is located at Asn77. 2013294 0 44 theme new 23:25 arg1 protein 41:47 a new actin-binding protein 21:47 a new actin-binding protein from human seminal plasma 21:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 7 45 theme fucose 693:698 arg1 fucose 693:698 fucose 693:698 fucose 693:698 The carbohydrate composition shows an unusually high amount of fucose. 2013294 7 45 theme fucose 693:698 arg1 amount 683:688 an unusually high amount 665:688 an unusually high amount of fucose 665:698 The carbohydrate composition shows an unusually high amount of fucose. 2013294 9 46 theme 64 904:905 arg1 % 906:906 % 906:906 % 906:906 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 10 47 dep protein 1022:1028 arg1 sequences 1034:1042 sequences 1034:1042 sequences translated from cDNA clones 1034:1070 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 5 48 theme SABP 457:460 arg1 chain 448:452 The single polypeptide chain 425:452 The single polypeptide chain of SABP 425:460 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 9 49 theme % 906:906 arg1 similarity 908:917 64% similarity 904:917 64% similarity 904:917 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 10 50 theme cDNA 1060:1063 arg1 clones 1065:1070 cDNA clones 1060:1070 cDNA clones 1060:1070 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 9 51 theme high 807:810 arg1 degree 812:817 a high degree 805:817 a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 805:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 0 52 theme protein 41:47 arg1 structure 8:16 Primary structure 0:16 Primary structure of a new actin-binding protein from human seminal plasma 0:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 1 53 gly glycoprotein 118:129 arg1 protein 100:106 Secretory actin-binding protein 76:106 Secretory actin-binding protein (SABP) 76:113 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 1 53 gly glycoprotein 118:129 arg1 glycoprotein 118:129 a glycoprotein 116:129 a glycoprotein from human seminal plasma 116:155 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 10 54 theme prolactin-inducible 946:964 arg1 protein 966:972 a prolactin-inducible protein 944:972 a prolactin-inducible protein 944:972 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 4 55 theme complete 279:286 arg1 sequence 299:306 The complete amino acid sequence 275:306 The complete amino acid sequence of SABP 275:314 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 2013294 8 56 theme disulfide 728:736 arg1 bonds 738:742 the two disulfide bonds 720:742 the two disulfide bonds 720:742 The arrangement of the two disulfide bonds is Cys37-Cys63 and Cys61-Cys95. 2013294 5 57 theme amino 475:479 arg1 residue 558:564 the N-terminal residue 543:564 the N-terminal residue 543:564 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 57 theme amino 475:479 arg1 acids 481:485 118 amino acids 471:485 118 amino acids with a calculated Mr of 13,506 471:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 57 theme amino 475:479 arg1 acid 535:538 pyroglutamic acid 522:538 pyroglutamic acid 522:538 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 58 contain contains 462:469 arg1 chain 448:452 The single polypeptide chain 425:452 The single polypeptide chain of SABP 425:460 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 58 contain contains 462:469 arg2 acids 481:485 118 amino acids 471:485 118 amino acids with a calculated Mr of 13,506 471:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 58 contain contains 462:469 arg2 residue 558:564 the N-terminal residue 543:564 the N-terminal residue 543:564 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 58 contain contains 462:469 arg2 acid 535:538 pyroglutamic acid 522:538 pyroglutamic acid 522:538 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 9 59 theme similarity 822:831 arg1 degree 812:817 a high degree 805:817 a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity) 805:918 Sequence comparison revealed a high degree of similarity with a 14-kDa submandibular gland protein from mouse (45% identity and 64% similarity). 2013294 5 60 theme single 429:434 arg1 chain 448:452 The single polypeptide chain 425:452 The single polypeptide chain of SABP 425:460 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 5 61 with acid 535:538 arg1 Mr 505:506 a calculated Mr 492:506 a calculated Mr of 13,506 492:516 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 7 62 theme high 678:681 arg1 fucose 693:698 fucose 693:698 fucose 693:698 The carbohydrate composition shows an unusually high amount of fucose. 2013294 7 62 theme high 678:681 arg1 amount 683:688 an unusually high amount 665:688 an unusually high amount of fucose 665:698 The carbohydrate composition shows an unusually high amount of fucose. 2013294 0 63 theme seminal 60:66 arg1 plasma 68:73 human seminal plasma 54:73 human seminal plasma 54:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 5 64 theme polypeptide 436:446 arg1 chain 448:452 The single polypeptide chain 425:452 The single polypeptide chain of SABP 425:460 The single polypeptide chain of SABP contains 118 amino acids with a calculated Mr of 13,506 and pyroglutamic acid as the N-terminal residue. 2013294 8 65 theme bonds 738:742 arg1 arrangement 705:715 The arrangement 701:715 The arrangement of the two disulfide bonds 701:742 The arrangement of the two disulfide bonds is Cys37-Cys63 and Cys61-Cys95. 2013294 8 65 theme bonds 738:742 arg1 Cys37-Cys63 747:757 Cys37-Cys63 747:757 Cys37-Cys63 747:757 The arrangement of the two disulfide bonds is Cys37-Cys63 and Cys61-Cys95. 2013294 0 66 theme human 54:58 arg1 plasma 68:73 human seminal plasma 54:73 human seminal plasma 54:73 Primary structure of a new actin-binding protein from human seminal plasma. 2013294 11 67 theme submandibular 1168:1180 arg1 gland 1182:1186 the submandibular gland 1164:1186 the submandibular gland 1164:1186 Although SABP was also detected in saliva, in extracts of the submandibular gland and seminal vesicles, little is known of its function. 2013294 1 68 from plasma 150:155 arg1 protein 100:106 Secretory actin-binding protein 76:106 Secretory actin-binding protein (SABP) 76:113 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 1 68 from plasma 150:155 arg1 glycoprotein 118:129 a glycoprotein 116:129 a glycoprotein from human seminal plasma 116:155 Secretory actin-binding protein (SABP), a glycoprotein from human seminal plasma, was isolated according to Akiyama and Kimura [Akiyama, K. & Kimura, H. (1990) Biochim. 2013294 10 69 with identical 929:937 arg1 both 1074:1077 both 1074:1077 both 1074:1077 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 10 69 with identical 929:937 arg1 protein 966:972 a prolactin-inducible protein 944:972 a prolactin-inducible protein 944:972 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 10 69 with identical 929:937 arg1 protein 980:986 a protein 978:986 a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones) 978:1071 SABP is identical with a prolactin-inducible protein and a protein termed gross cystic disease fluid protein 15 (sequences translated from cDNA clones), both from human breast tissues. 2013294 4 70 dep the 336:338 arg1 aid 340:342 aid 340:342 aid 340:342 The complete amino acid sequence of SABP was determined with the aid of fragments generated by trypsin, Staphylococcus aureus V8 protease and pepsin. 18764929 8 0 theme disease 1679:1685 arg1 implications 1687:1698 possible disease implications 1670:1698 possible disease implications 1670:1698 These findings point to novel intracellular functions for beta-DG, with possible disease implications. 18764929 3 1 theme SEA 530:532 arg1 domain 535:540 sea urchin-enterokinase-agrin (SEA) domain 499:540 sea urchin-enterokinase-agrin (SEA) domain 499:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 3 2 theme heterodimer 580:590 arg1 creation 592:599 this cleavage and heterodimer creation 562:599 creation 592:599 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 6 3 theme DG 1188:1189 arg1 subunits 1162:1169 the transmembrane subunits 1144:1169 the transmembrane subunits of both MUC1 and DG 1144:1189 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 7 4 theme nuclear 1500:1506 arg1 localization 1508:1519 nuclear localization 1500:1519 nuclear localization of beta-DG 1500:1530 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 7 5 theme nuclear 1451:1457 arg1 partitioning 1459:1470 the nuclear partitioning 1447:1470 the nuclear partitioning of beta-DG 1447:1481 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 4 6 theme nuclear 655:661 arg1 signal 676:681 a functional nuclear localization signal 642:681 a functional nuclear localization signal 642:681 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 2 7 theme extracellular 282:294 arg1 subunits 335:342 extracellular (alpha-DG) and transmembrane (beta-DG) subunits 282:342 extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface 282:377 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 18764929 9 8 theme SEA 1755:1757 arg1 autoproteolysis 1759:1773 SEA autoproteolysis 1755:1773 SEA autoproteolysis 1755:1773 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 6 9 theme prior 1221:1225 arg1 pathway 1213:1219 the secretory pathway 1199:1219 the secretory pathway prior to nuclear targeting 1199:1246 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 7 10 theme considerable 1419:1430 arg1 variability 1432:1442 considerable variability 1419:1442 considerable variability in the nuclear partitioning of beta-DG 1419:1481 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 7 11 theme origins 1402:1408 arg1 lines 1365:1369 breast carcinoma cell lines 1343:1369 breast carcinoma cell lines of distinct pathophysiological origins 1343:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 5 12 theme membrane-tethered 1075:1091 arg1 mucin 1093:1097 a heterodimeric membrane-tethered mucin 1059:1097 a heterodimeric membrane-tethered mucin 1059:1097 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 2 13 theme mucin 250:254 arg1 family 256:261 the membrane-tethered mucin family 228:261 the membrane-tethered mucin family 228:261 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 18764929 7 14 theme distinct 1374:1381 arg1 origins 1402:1408 distinct pathophysiological origins 1374:1408 distinct pathophysiological origins 1374:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 2 15 dep transmembrane 311:323 arg1 beta-DG 326:332 beta-DG 326:332 beta-DG 326:332 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 18764929 5 16 theme MUC1 1036:1039 arg1 oncoprotein 1041:1051 the related MUC1 oncoprotein 1024:1051 the related MUC1 oncoprotein 1024:1051 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 3 17 theme creation 592:599 arg1 uncertain 604:612 uncertain 604:612 uncertain 604:612 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 3 17 theme creation 592:599 arg1 purpose 551:557 the purpose 547:557 the purpose of this cleavage and heterodimer creation 547:599 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 9 18 theme independent 1794:1804 arg1 functions 1806:1814 independent functions 1794:1814 independent functions 1794:1814 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 7 19 theme lines 1365:1369 arg1 screen 1333:1338 A screen 1331:1338 A screen of breast carcinoma cell lines of distinct pathophysiological origins 1331:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 1 20 theme extracellular 136:148 arg1 receptor 157:164 an extracellular matrix receptor 133:164 an extracellular matrix receptor implicated in muscular dystrophies and cancers 133:211 Dystroglycan (DG) is an extracellular matrix receptor implicated in muscular dystrophies and cancers. 18764929 1 20 theme extracellular 136:148 arg1 Dystroglycan 112:123 Dystroglycan 112:123 Dystroglycan (DG) 112:128 Dystroglycan (DG) is an extracellular matrix receptor implicated in muscular dystrophies and cancers. 18764929 7 21 theme carcinoma 1350:1358 arg1 lines 1365:1369 breast carcinoma cell lines 1343:1369 breast carcinoma cell lines of distinct pathophysiological origins 1343:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 9 22 theme transmembrane 1825:1837 arg1 subunits 1839:1846 mucin transmembrane subunits 1819:1846 mucin transmembrane subunits 1819:1846 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 0 23 theme autoproteolyzed 88:102 arg1 mucins 104:109 autoproteolyzed mucins 88:109 autoproteolyzed mucins 88:109 Nuclear translocation of beta-dystroglycan reveals a distinctive trafficking pattern of autoproteolyzed mucins. 18764929 3 24 theme monomeric 437:445 arg1 precursor 447:455 a monomeric precursor 435:455 a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain 435:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 5 25 theme SEA 1110:1112 arg1 autoproteolysis 1114:1128 SEA autoproteolysis 1110:1128 SEA autoproteolysis 1110:1128 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 6 26 theme transmembrane 1148:1160 arg1 subunits 1162:1169 the transmembrane subunits 1144:1169 the transmembrane subunits of both MUC1 and DG 1144:1189 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 7 27 theme breast 1343:1348 arg1 lines 1365:1369 breast carcinoma cell lines 1343:1369 breast carcinoma cell lines of distinct pathophysiological origins 1343:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 9 28 theme shared 1862:1867 arg1 pathway 1891:1897 a shared and poorly understood pathway 1860:1897 a shared and poorly understood pathway of segregated subunit trafficking 1860:1931 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 0 29 theme Nuclear 0:6 arg1 translocation 8:20 Nuclear translocation 0:20 Nuclear translocation of beta-dystroglycan 0:41 Nuclear translocation of beta-dystroglycan reveals a distinctive trafficking pattern of autoproteolyzed mucins. 18764929 5 30 theme trafficking 940:950 arg1 pattern 952:958 The trafficking pattern 936:958 The trafficking pattern of beta-DG 936:969 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 3 31 theme distinctive 466:476 arg1 motif 486:490 a distinctive protein motif 464:490 a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain 464:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 6 32 theme MUC1 1179:1182 arg1 subunits 1162:1169 the transmembrane subunits 1144:1169 the transmembrane subunits of both MUC1 and DG 1144:1189 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 7 33 theme ligand 1582:1587 arg1 binding 1589:1595 extracellular ligand binding 1568:1595 extracellular ligand binding 1568:1595 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 9 34 theme segregated 1902:1911 arg1 trafficking 1921:1931 segregated subunit trafficking 1902:1931 segregated subunit trafficking 1902:1931 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 3 35 theme sea 499:501 arg1 domain 535:540 sea urchin-enterokinase-agrin (SEA) domain 499:540 sea urchin-enterokinase-agrin (SEA) domain 499:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 0 36 theme distinctive 53:63 arg1 pattern 77:83 a distinctive trafficking pattern 51:83 a distinctive trafficking pattern of autoproteolyzed mucins 51:109 Nuclear translocation of beta-dystroglycan reveals a distinctive trafficking pattern of autoproteolyzed mucins. 18764929 5 37 theme related 1028:1034 arg1 oncoprotein 1041:1051 the related MUC1 oncoprotein 1024:1051 the related MUC1 oncoprotein 1024:1051 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 9 38 theme trafficking 1921:1931 arg1 pathway 1891:1897 a shared and poorly understood pathway 1860:1897 a shared and poorly understood pathway of segregated subunit trafficking 1860:1931 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 8 39 theme intracellular 1628:1640 arg1 functions 1642:1650 novel intracellular functions 1622:1650 novel intracellular functions for beta-DG 1622:1662 These findings point to novel intracellular functions for beta-DG, with possible disease implications. 18764929 7 40 theme beta-DG 1524:1530 arg1 localization 1508:1519 nuclear localization 1500:1519 nuclear localization of beta-DG 1500:1530 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 5 41 theme transmembrane 999:1011 arg1 MUC1-C 987:992 MUC1-C 987:992 MUC1-C 987:992 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 5 41 theme transmembrane 999:1011 arg1 subunit 1013:1019 the transmembrane subunit 995:1019 the transmembrane subunit of the related MUC1 oncoprotein 995:1051 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 8 42 theme possible 1670:1677 arg1 implications 1687:1698 possible disease implications 1670:1698 possible disease implications 1670:1698 These findings point to novel intracellular functions for beta-DG, with possible disease implications. 18764929 7 43 theme beta-DG 1475:1481 arg1 partitioning 1459:1470 the nuclear partitioning 1447:1470 the nuclear partitioning of beta-DG 1447:1481 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 6 44 theme monomeric 1263:1271 arg1 precursors 1273:1282 their monomeric precursors 1257:1282 their monomeric precursors 1257:1282 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 1 45 theme muscular 180:187 arg1 dystrophies 189:199 muscular dystrophies 180:199 muscular dystrophies 180:199 Dystroglycan (DG) is an extracellular matrix receptor implicated in muscular dystrophies and cancers. 18764929 9 46 theme conserved 1736:1744 arg1 role 1746:1749 an evolutionarily conserved role 1718:1749 an evolutionarily conserved role for SEA autoproteolysis 1718:1773 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 4 47 theme functional 644:653 arg1 signal 676:681 a functional nuclear localization signal 642:681 a functional nuclear localization signal 642:681 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 3 48 theme cleavage 567:574 arg1 creation 592:599 this cleavage and heterodimer creation 562:599 creation 592:599 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 4 49 theme localization 663:674 arg1 signal 676:681 a functional nuclear localization signal 642:681 a functional nuclear localization signal 642:681 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 2 50 theme transmembrane 311:323 arg1 subunits 335:342 extracellular (alpha-DG) and transmembrane (beta-DG) subunits 282:342 extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface 282:377 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 18764929 4 51 theme alpha-DG 911:918 arg1 binding 927:933 alpha-DG ligand binding 911:933 alpha-DG ligand binding 911:933 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 5 52 theme heterodimeric 1061:1073 arg1 mucin 1093:1097 a heterodimeric membrane-tethered mucin 1059:1097 a heterodimeric membrane-tethered mucin 1059:1097 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 4 53 theme binding 927:933 arg1 independent 896:906 independent 896:906 independent 896:906 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 6 54 theme nuclear 1230:1236 arg1 targeting 1238:1246 nuclear targeting 1230:1246 nuclear targeting 1230:1246 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 2 55 theme membrane-tethered 232:248 arg1 family 256:261 the membrane-tethered mucin family 228:261 the membrane-tethered mucin family 228:261 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 18764929 4 56 theme ligand 920:925 arg1 binding 927:933 alpha-DG ligand binding 911:933 alpha-DG ligand binding 911:933 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 7 57 theme pathophysiological 1383:1400 arg1 origins 1402:1408 distinct pathophysiological origins 1374:1408 distinct pathophysiological origins 1374:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 6 58 theme nuclear 1310:1316 arg1 trafficking 1318:1328 nuclear trafficking 1310:1328 nuclear trafficking 1310:1328 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 5 59 theme oncoprotein 1041:1051 arg1 MUC1-C 987:992 MUC1-C 987:992 MUC1-C 987:992 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 5 59 theme oncoprotein 1041:1051 arg1 subunit 1013:1019 the transmembrane subunit 995:1019 the transmembrane subunit of the related MUC1 oncoprotein 995:1051 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 4 60 dep associating 728:738 arg1 addition 716:723 addition 716:723 addition 716:723 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 7 61 from variability 1432:1442 arg1 partitioning 1459:1470 the nuclear partitioning 1447:1470 the nuclear partitioning of beta-DG 1447:1481 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 6 62 theme secretory 1203:1211 arg1 pathway 1213:1219 the secretory pathway 1199:1219 the secretory pathway prior to nuclear targeting 1199:1246 We show that the transmembrane subunits of both MUC1 and DG transit the secretory pathway prior to nuclear targeting and that their monomeric precursors maintain the capacity for nuclear trafficking. 18764929 1 63 theme matrix 150:155 arg1 receptor 157:164 an extracellular matrix receptor 133:164 an extracellular matrix receptor implicated in muscular dystrophies and cancers 133:211 Dystroglycan (DG) is an extracellular matrix receptor implicated in muscular dystrophies and cancers. 18764929 1 63 theme matrix 150:155 arg1 Dystroglycan 112:123 Dystroglycan 112:123 Dystroglycan (DG) 112:128 Dystroglycan (DG) is an extracellular matrix receptor implicated in muscular dystrophies and cancers. 18764929 9 64 theme mucin 1819:1823 arg1 subunits 1839:1846 mucin transmembrane subunits 1819:1846 mucin transmembrane subunits 1819:1846 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 7 65 theme cell 1360:1363 arg1 lines 1365:1369 breast carcinoma cell lines 1343:1369 breast carcinoma cell lines of distinct pathophysiological origins 1343:1408 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 2 66 dep extracellular 282:294 arg1 alpha-DG 297:304 alpha-DG 297:304 alpha-DG 297:304 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 18764929 0 67 theme mucins 104:109 arg1 pattern 77:83 a distinctive trafficking pattern 51:83 a distinctive trafficking pattern of autoproteolyzed mucins 51:109 Nuclear translocation of beta-dystroglycan reveals a distinctive trafficking pattern of autoproteolyzed mucins. 18764929 9 68 theme subunits 1839:1846 arg1 functions 1806:1814 independent functions 1794:1814 independent functions 1794:1814 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 4 69 from nucleoplasm 859:869 arg1 process 876:882 a process 874:882 a process that occurs independent of alpha-DG ligand binding 874:933 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 5 70 theme beta-DG 963:969 arg1 pattern 952:958 The trafficking pattern 936:958 The trafficking pattern of beta-DG 936:969 The trafficking pattern of beta-DG mirrors that of MUC1-C, the transmembrane subunit of the related MUC1 oncoprotein, also a heterodimeric membrane-tethered mucin created by SEA autoproteolysis. 18764929 4 71 from cytoplasm 845:853 arg1 process 876:882 a process 874:882 a process that occurs independent of alpha-DG ligand binding 874:933 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 4 72 theme cell 761:764 arg1 surface 766:772 the cell surface 757:772 the cell surface 757:772 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 3 73 theme precursor 447:455 arg1 autoproteolysis 416:430 autoproteolysis 416:430 autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain 416:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 4 74 gly glycosylated 795:806 arg1 beta-DG 808:814 the full-length and glycosylated beta-DG 775:814 the full-length and glycosylated beta-DG 775:814 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 0 75 theme beta-dystroglycan 25:41 arg1 translocation 8:20 Nuclear translocation 0:20 Nuclear translocation of beta-dystroglycan 0:41 Nuclear translocation of beta-dystroglycan reveals a distinctive trafficking pattern of autoproteolyzed mucins. 18764929 9 76 theme understood 1880:1889 arg1 pathway 1891:1897 a shared and poorly understood pathway 1860:1897 a shared and poorly understood pathway of segregated subunit trafficking 1860:1931 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 3 77 theme protein 478:484 arg1 motif 486:490 a distinctive protein motif 464:490 a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain 464:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 7 78 theme binding 1589:1595 arg1 independent 1553:1563 independent 1553:1563 independent 1553:1563 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 8 79 theme novel 1622:1626 arg1 functions 1642:1650 novel intracellular functions 1622:1650 novel intracellular functions for beta-DG 1622:1662 These findings point to novel intracellular functions for beta-DG, with possible disease implications. 18764929 4 80 theme glycosylated 795:806 arg1 beta-DG 808:814 the full-length and glycosylated beta-DG 775:814 the full-length and glycosylated beta-DG 775:814 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 0 81 theme trafficking 65:75 arg1 pattern 77:83 a distinctive trafficking pattern 51:83 a distinctive trafficking pattern of autoproteolyzed mucins 51:109 Nuclear translocation of beta-dystroglycan reveals a distinctive trafficking pattern of autoproteolyzed mucins. 18764929 7 82 theme extracellular 1568:1580 arg1 binding 1589:1595 extracellular ligand binding 1568:1595 extracellular ligand binding 1568:1595 A screen of breast carcinoma cell lines of distinct pathophysiological origins revealed considerable variability in the nuclear partitioning of beta-DG, indicating that nuclear localization of beta-DG is regulated, albeit independent of extracellular ligand binding. 18764929 9 83 theme subunit 1913:1919 arg1 trafficking 1921:1931 segregated subunit trafficking 1902:1931 segregated subunit trafficking 1902:1931 They also reveal an evolutionarily conserved role for SEA autoproteolysis, serving to enable independent functions of mucin transmembrane subunits, enacted by a shared and poorly understood pathway of segregated subunit trafficking. 18764929 3 84 theme urchin-enterokinase-agrin 503:527 arg1 domain 535:540 sea urchin-enterokinase-agrin (SEA) domain 499:540 sea urchin-enterokinase-agrin (SEA) domain 499:540 These two subunits are generated by autoproteolysis of a monomeric precursor within a distinctive protein motif called sea urchin-enterokinase-agrin (SEA) domain, yet the purpose of this cleavage and heterodimer creation is uncertain. 18764929 4 85 dep cytoplasm 845:853 arg1 the 841:843 the 841:843 the 841:843 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 4 86 theme full-length 779:789 arg1 beta-DG 808:814 the full-length and glycosylated beta-DG 775:814 the full-length and glycosylated beta-DG 775:814 In this study, we identify a functional nuclear localization signal within beta-DG and show that, in addition to associating with alpha-DG at the cell surface, the full-length and glycosylated beta-DG autonomously traffics to the cytoplasm and nucleoplasm in a process that occurs independent of alpha-DG ligand binding. 18764929 2 87 theme cell 366:369 arg1 surface 371:377 the cell surface 362:377 the cell surface 362:377 DG belongs to the membrane-tethered mucin family and is composed of extracellular (alpha-DG) and transmembrane (beta-DG) subunits stably coupled at the cell surface. 21148085 4 0 from resolution 618:627 arg1 structure 562:570 the crystal structure 550:570 the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution 550:627 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 5 1 theme VEGF-C 659:664 arg1 Comparison 630:639 Comparison 630:639 Comparison of the VEGF-D and VEGF-C structures 630:675 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 1 2 theme kinase 161:166 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 2 theme kinase 161:166 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 2 theme kinase 161:166 arg1 VEGFR-1-3 179:187 VEGFR-1-3 179:187 VEGFR-1-3 179:187 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 2 theme kinase 161:166 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 4 3 theme mutant 602:607 arg1 structure 562:570 the crystal structure 550:570 the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution 550:627 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 5 4 dep overall 730:736 arg1 folds 738:742 folds 738:742 folds 738:742 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 8 5 theme VEGF-D 1091:1096 arg1 polypeptides 1098:1109 N-terminally truncated VEGF-D polypeptides 1068:1109 N-terminally truncated VEGF-D polypeptides 1068:1109 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 8 6 theme functional 1047:1056 arg1 assays 1058:1063 functional assays 1047:1063 functional assays 1047:1063 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 2 7 with proteins 311:318 arg1 propeptides 348:358 C-terminal propeptides 337:358 C-terminal propeptides 337:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 7 with proteins 311:318 arg1 N- 330:331 long N- 325:331 long N- 325:331 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 9 8 theme VEGF-D 1315:1320 arg1 angiogenic 1330:1339 angiogenic 1330:1339 angiogenic 1330:1339 Thus, we define here a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic. 21148085 9 8 theme VEGF-D 1315:1320 arg1 form 1307:1310 a VEGFR-2-specific form 1288:1310 a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic 1288:1363 Thus, we define here a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic. 21148085 10 9 theme VEGF-D 1416:1421 arg1 structure 1423:1431 VEGF-D structure 1416:1431 VEGF-D structure 1416:1431 These results provide important new insights into VEGF-D structure and function. 21148085 2 10 theme precursor 301:309 arg1 ligands 259:265 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 10 theme precursor 301:309 arg1 proteins 311:318 precursor proteins 301:318 precursor proteins with long N- and C-terminal propeptides 301:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 10 theme precursor 301:309 arg1 binding 398:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 7 11 theme VEGF-C 904:909 arg1 structures 911:920 VEGF-C structures 904:920 VEGF-C structures 904:920 In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. 21148085 5 12 theme VEGFR-2 749:755 arg1 residues 769:776 VEGFR-2 interacting residues 749:776 VEGFR-2 interacting residues 749:776 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 5 13 theme similar 683:689 arg1 helices 711:717 similar extended N-terminal helices 683:717 similar extended N-terminal helices 683:717 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 6 14 theme thermodynamic 822:834 arg1 parameters 836:845 the thermodynamic parameters 818:845 the thermodynamic parameters for VEGFR-2 binding 818:865 Consistent with this, the affinity and the thermodynamic parameters for VEGFR-2 binding are very similar. 21148085 6 14 theme thermodynamic 822:834 arg1 similar 876:882 similar 876:882 similar 876:882 Consistent with this, the affinity and the thermodynamic parameters for VEGFR-2 binding are very similar. 21148085 2 15 theme enhanced 369:376 arg1 binding 398:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 15 theme enhanced 369:376 arg1 proteins 311:318 precursor proteins 301:318 precursor proteins with long N- and C-terminal propeptides 301:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 15 theme enhanced 369:376 arg1 ligands 259:265 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 7 16 theme better 1002:1007 arg1 resolution 1009:1018 a better resolution 1000:1018 a better resolution 1000:1018 In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. 21148085 5 17 theme interacting 757:767 arg1 residues 769:776 VEGFR-2 interacting residues 749:776 VEGFR-2 interacting residues 749:776 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 8 18 theme N-terminally 1068:1079 arg1 polypeptides 1098:1109 N-terminally truncated VEGF-D polypeptides 1068:1109 N-terminally truncated VEGF-D polypeptides 1068:1109 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 2 19 theme proteolytic 409:419 arg1 removal 421:427 proteolytic removal 409:427 proteolytic removal of the propeptides 409:446 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 8 20 theme VEGF-D 1205:1210 arg1 binding 1212:1218 VEGF-D binding 1205:1218 VEGF-D binding 1205:1218 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 5 21 dep VEGF-D 648:653 arg1 structures 666:675 structures 666:675 structures 666:675 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 5 21 dep VEGF-D 648:653 arg1 the 644:646 the 644:646 the 644:646 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 4 22 theme Å 616:616 arg1 resolution 618:627 2.9 Å resolution 612:627 2.9 Å resolution 612:627 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 2 23 dep ligands 259:265 arg1 VEGF-C 267:272 VEGF-C 267:272 VEGF-C 267:272 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 23 dep ligands 259:265 arg1 ligands 259:265 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 23 dep ligands 259:265 arg1 VEGF-D 278:283 VEGF-D 278:283 VEGF-D 278:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 5 24 theme VEGF-D 648:653 arg1 Comparison 630:639 Comparison 630:639 Comparison of the VEGF-D and VEGF-C structures 630:675 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 8 25 theme cleavage 1172:1179 arg1 sites 1181:1185 the reported proteolytic cleavage sites 1147:1185 the reported proteolytic cleavage sites 1147:1185 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 1 26 theme Vascular 98:105 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 26 theme Vascular 98:105 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 26 theme Vascular 98:105 arg1 VEGFs 135:139 VEGFs 135:139 VEGFs 135:139 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 26 theme Vascular 98:105 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 0 27 theme Structural 0:9 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of vascular endothelial growth factor-D 0:62 Structural determinants of vascular endothelial growth factor-D receptor binding and specificity. 21148085 7 28 with comparison 888:897 arg1 structures 911:920 VEGF-C structures 904:920 VEGF-C structures 904:920 In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. 21148085 7 29 theme N-terminal 943:952 arg1 helix 954:958 the VEGF-D N-terminal helix 932:958 the VEGF-D N-terminal helix 932:958 In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. 21148085 1 30 theme endothelial 107:117 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 30 theme endothelial 107:117 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 30 theme endothelial 107:117 arg1 VEGFs 135:139 VEGFs 135:139 VEGFs 135:139 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 30 theme endothelial 107:117 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 2 31 theme VEGFR-3 390:396 arg1 binding 398:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 31 theme VEGFR-3 390:396 arg1 proteins 311:318 precursor proteins 301:318 precursor proteins with long N- and C-terminal propeptides 301:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 31 theme VEGFR-3 390:396 arg1 ligands 259:265 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 0 32 theme vascular 27:34 arg1 factor-D 55:62 vascular endothelial growth factor-D 27:62 vascular endothelial growth factor-D 27:62 Structural determinants of vascular endothelial growth factor-D receptor binding and specificity. 21148085 8 33 theme receptor 1026:1033 arg1 binding 1035:1041 receptor binding 1026:1041 receptor binding 1026:1041 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 8 34 theme truncated 1081:1089 arg1 polypeptides 1098:1109 N-terminally truncated VEGF-D polypeptides 1068:1109 N-terminally truncated VEGF-D polypeptides 1068:1109 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 1 35 theme growth 119:124 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 35 theme growth 119:124 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 35 theme growth 119:124 arg1 VEGFs 135:139 VEGFs 135:139 VEGFs 135:139 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 35 theme growth 119:124 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 2 36 theme propeptides 436:446 arg1 removal 421:427 proteolytic removal 409:427 proteolytic removal of the propeptides 409:446 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 37 theme long 325:328 arg1 N- 330:331 long N- 325:331 long N- 325:331 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 4 38 theme VEGF-D 585:590 arg1 mutant 602:607 the human VEGF-D Cys117Ala mutant 575:607 the human VEGF-D Cys117Ala mutant 575:607 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 8 39 theme polypeptides 1098:1109 arg1 binding 1035:1041 receptor binding 1026:1041 receptor binding 1026:1041 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 8 39 theme polypeptides 1098:1109 arg1 assays 1058:1063 functional assays 1047:1063 functional assays 1047:1063 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 9 40 theme VEGFR-2-specific 1290:1305 arg1 angiogenic 1330:1339 angiogenic 1330:1339 angiogenic 1330:1339 Thus, we define here a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic. 21148085 9 40 theme VEGFR-2-specific 1290:1305 arg1 form 1307:1310 a VEGFR-2-specific form 1288:1310 a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic 1288:1363 Thus, we define here a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic. 21148085 0 41 theme growth 48:53 arg1 factor-D 55:62 vascular endothelial growth factor-D 27:62 vascular endothelial growth factor-D 27:62 Structural determinants of vascular endothelial growth factor-D receptor binding and specificity. 21148085 4 42 theme human 579:583 arg1 mutant 602:607 the human VEGF-D Cys117Ala mutant 575:607 the human VEGF-D Cys117Ala mutant 575:607 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 2 43 theme VEGFR-3 251:257 arg1 VEGF-C 267:272 VEGF-C 267:272 VEGF-C 267:272 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 43 theme VEGFR-3 251:257 arg1 binding 398:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 43 theme VEGFR-3 251:257 arg1 proteins 311:318 precursor proteins 301:318 precursor proteins with long N- and C-terminal propeptides 301:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 43 theme VEGFR-3 251:257 arg1 ligands 259:265 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 43 theme VEGFR-3 251:257 arg1 VEGF-D 278:283 VEGF-D 278:283 VEGF-D 278:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 3 44 theme cleavage 475:482 arg1 sites 484:488 Two different proteolytic cleavage sites 449:488 Two different proteolytic cleavage sites 449:488 Two different proteolytic cleavage sites have been reported in the VEGF-D N-terminus. 21148085 0 45 theme endothelial 36:46 arg1 factor-D 55:62 vascular endothelial growth factor-D 27:62 vascular endothelial growth factor-D 27:62 Structural determinants of vascular endothelial growth factor-D receptor binding and specificity. 21148085 10 46 theme important 1388:1396 arg1 insights 1402:1409 important new insights 1388:1409 important new insights into VEGF-D structure and function 1388:1444 These results provide important new insights into VEGF-D structure and function. 21148085 4 47 theme Cys117Ala 592:600 arg1 mutant 602:607 the human VEGF-D Cys117Ala mutant 575:607 the human VEGF-D Cys117Ala mutant 575:607 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 8 48 theme VEGFR-3 1238:1244 arg1 activation 1224:1233 activation 1224:1233 activation of VEGFR-3 1224:1244 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 1 49 theme angiogenesis 215:226 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 49 theme angiogenesis 215:226 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 49 theme angiogenesis 215:226 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 5 50 theme extended 691:698 arg1 helices 711:717 similar extended N-terminal helices 683:717 similar extended N-terminal helices 683:717 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 3 51 theme different 453:461 arg1 sites 484:488 Two different proteolytic cleavage sites 449:488 Two different proteolytic cleavage sites 449:488 Two different proteolytic cleavage sites have been reported in the VEGF-D N-terminus. 21148085 0 52 theme factor-D 55:62 arg1 determinants 11:22 Structural determinants 0:22 Structural determinants of vascular endothelial growth factor-D 0:62 Structural determinants of vascular endothelial growth factor-D receptor binding and specificity. 21148085 5 53 theme N-terminal 700:709 arg1 helices 711:717 similar extended N-terminal helices 683:717 similar extended N-terminal helices 683:717 Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. 21148085 3 54 theme VEGF-D 516:521 arg1 N-terminus 523:532 the VEGF-D N-terminus 512:532 the VEGF-D N-terminus 512:532 Two different proteolytic cleavage sites have been reported in the VEGF-D N-terminus. 21148085 4 55 theme crystal 554:560 arg1 structure 562:570 the crystal structure 550:570 the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution 550:627 We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. 21148085 8 56 theme reported 1151:1158 arg1 sites 1181:1185 the reported proteolytic cleavage sites 1147:1185 the reported proteolytic cleavage sites 1147:1185 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 2 57 theme C-terminal 337:346 arg1 propeptides 348:358 C-terminal propeptides 337:358 C-terminal propeptides 337:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 7 58 theme VEGF-D 936:941 arg1 helix 954:958 the VEGF-D N-terminal helix 932:958 the VEGF-D N-terminal helix 932:958 In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. 21148085 10 59 theme new 1398:1400 arg1 insights 1402:1409 important new insights 1388:1409 important new insights into VEGF-D structure and function 1388:1444 These results provide important new insights into VEGF-D structure and function. 21148085 1 60 theme tyrosine 152:159 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 60 theme tyrosine 152:159 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 60 theme tyrosine 152:159 arg1 VEGFR-1-3 179:187 VEGFR-1-3 179:187 VEGFR-1-3 179:187 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 60 theme tyrosine 152:159 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 61 theme lymphangiogenesis 232:248 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 61 theme lymphangiogenesis 232:248 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 61 theme lymphangiogenesis 232:248 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 6 62 with Consistent 779:788 arg1 this 795:798 this 795:798 this 795:798 Consistent with this, the affinity and the thermodynamic parameters for VEGFR-2 binding are very similar. 21148085 6 63 theme VEGFR-2 851:857 arg1 binding 859:865 VEGFR-2 binding 851:865 VEGFR-2 binding 851:865 Consistent with this, the affinity and the thermodynamic parameters for VEGFR-2 binding are very similar. 21148085 1 64 theme central 194:200 arg1 mediators 202:210 central mediators 194:210 central mediators of angiogenesis and lymphangiogenesis 194:248 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 64 theme central 194:200 arg1 receptors 168:176 their tyrosine kinase receptors 146:176 their tyrosine kinase receptors (VEGFR-1-3) 146:188 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 1 64 theme central 194:200 arg1 factors 126:132 Vascular endothelial growth factors 98:132 Vascular endothelial growth factors (VEGFs) 98:140 Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. 21148085 3 65 theme proteolytic 463:473 arg1 sites 484:488 Two different proteolytic cleavage sites 449:488 Two different proteolytic cleavage sites 449:488 Two different proteolytic cleavage sites have been reported in the VEGF-D N-terminus. 21148085 8 66 theme proteolytic 1160:1170 arg1 cleavage 1172:1179 proteolytic cleavage 1160:1179 the reported proteolytic cleavage sites 1147:1185 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 8 67 theme VEGFR-2 1258:1264 arg1 important 1191:1199 important 1191:1199 important 1191:1199 Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. 21148085 2 68 theme VEGFR-2 378:384 arg1 binding 398:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 enhanced VEGFR-2 and VEGFR-3 binding 369:404 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 68 theme VEGFR-2 378:384 arg1 proteins 311:318 precursor proteins 301:318 precursor proteins with long N- and C-terminal propeptides 301:358 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 21148085 2 68 theme VEGFR-2 378:384 arg1 ligands 259:265 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D 251:283 VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. 17286803 0 0 theme endosomal 83:91 arg1 sorting 93:99 efficient endosomal sorting 73:99 efficient endosomal sorting to lysosomes 73:112 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 7 1 from level 831:835 arg1 lysosomes 848:856 lysosomes 848:856 lysosomes most likely 848:868 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 1 2 theme glycoprotein 161:172 arg1 CLN3 174:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 6 3 theme farnesyltransferase 677:695 arg1 inhibitor 697:705 farnesyltransferase inhibitor 677:705 farnesyltransferase inhibitor 677:705 Mevalonate incorporation and farnesyltransferase inhibitor studies indicate that CLN3 is prenylated most likely at cysteine 435. 17286803 7 4 theme cysteine 793:800 arg1 Substitution 777:788 Substitution 777:788 Substitution of cysteine 435 777:804 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 0 5 theme efficient 73:81 arg1 sorting 93:99 efficient endosomal sorting 73:99 efficient endosomal sorting to lysosomes 73:112 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 9 6 theme mannose 1240:1246 arg1 receptors 1260:1268 mannose 6-phosphate receptors 1240:1268 mannose 6-phosphate receptors 1240:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 7 theme 6-phosphate 1248:1258 arg1 receptors 1260:1268 mannose 6-phosphate receptors 1240:1268 mannose 6-phosphate receptors 1240:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 5 8 dep CLN3 602:605 arg1 Both 566:569 Both 566:569 Both 566:569 Both partially and non-glycosylated CLN3 were transported correctly to lysosomes. 17286803 5 8 dep CLN3 602:605 arg1 non-glycosylated 585:600 non-glycosylated 585:600 non-glycosylated 585:600 Both partially and non-glycosylated CLN3 were transported correctly to lysosomes. 17286803 3 9 theme motif 389:393 arg1 role 343:346 the role 339:346 the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells 339:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 4 10 theme asparagine 535:544 arg1 residues 546:553 asparagine residues 71 and 85 535:563 residues 546:553 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 3 11 theme CLN3 421:424 arg1 transport 408:416 lysosomal transport 398:416 lysosomal transport of CLN3 in non-neuronal and neuronal cells 398:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 6 12 theme incorporation 659:671 arg1 studies 707:713 Mevalonate incorporation and farnesyltransferase inhibitor studies 648:713 Mevalonate incorporation and farnesyltransferase inhibitor studies 648:713 Mevalonate incorporation and farnesyltransferase inhibitor studies indicate that CLN3 is prenylated most likely at cysteine 435. 17286803 9 13 theme endoplasmic 1377:1387 arg1 reticulum 1389:1397 the endoplasmic reticulum 1373:1397 the endoplasmic reticulum 1373:1397 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 10 14 theme cytoplasmic 1466:1476 arg1 CLN3 1489:1492 CLN3 1489:1492 CLN3 1489:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 10 14 theme cytoplasmic 1466:1476 arg1 domains 1478:1484 different cytoplasmic domains 1456:1484 different cytoplasmic domains of CLN3 1456:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 9 15 theme reticulum 1389:1397 arg1 exit 1365:1368 exit 1365:1368 exit of the endoplasmic reticulum 1365:1397 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 10 16 theme co-operative 1422:1433 arg1 structures 1442:1451 co-operative signal structures 1422:1451 co-operative signal structures in different cytoplasmic domains of CLN3 1422:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 8 17 theme inhibitors 1064:1073 arg1 presence 1032:1039 the presence 1028:1039 the presence of farnesyltransferase inhibitors 1028:1073 Additionally, the cell surface expression of CLN3 was increased in the presence of farnesyltransferase inhibitors. 17286803 8 18 theme cell 979:982 arg1 expression 992:1001 the cell surface expression 975:1001 the cell surface expression of CLN3 975:1009 Additionally, the cell surface expression of CLN3 was increased in the presence of farnesyltransferase inhibitors. 17286803 4 19 gly glycosylated 519:530 arg1 CLN3 511:514 CLN3 511:514 CLN3 511:514 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 4 19 gly glycosylated 519:530 arg2 residues 546:553 asparagine residues 71 and 85 535:563 residues 546:553 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 4 19 gly glycosylated 519:530 arg1 cells 504:508 COS7 cells 499:508 COS7 cells 499:508 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 4 19 gly glycosylated 519:530 arg2 CLN3 511:514 CLN3 511:514 CLN3 511:514 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 4 19 gly glycosylated 519:530 arg2 85 562:563 85 562:563 85 562:563 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 3 20 theme C-terminal 373:382 arg1 motif 389:393 the C-terminal CAAX motif 369:393 the C-terminal CAAX motif 369:393 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 3 21 from transport 408:416 arg1 cells 455:459 non-neuronal and neuronal cells 429:459 cells 455:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 8 22 theme farnesyltransferase 1044:1062 arg1 inhibitors 1064:1073 farnesyltransferase inhibitors 1044:1073 farnesyltransferase inhibitors 1044:1073 Additionally, the cell surface expression of CLN3 was increased in the presence of farnesyltransferase inhibitors. 17286803 3 23 theme CAAX 384:387 arg1 motif 389:393 the C-terminal CAAX motif 369:393 the C-terminal CAAX motif 369:393 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 9 24 with CLN3 1209:1212 arg1 tails 1231:1235 cytoplasmic tails 1219:1235 cytoplasmic tails of mannose 6-phosphate receptors 1219:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 2 25 theme lysosomal 302:310 arg1 targeting 312:320 lysosomal targeting 302:320 lysosomal targeting 302:320 Previous studies identified two cytosolic signal structures contributing to lysosomal targeting. 17286803 5 26 dep Both 566:569 arg1 partially 571:579 partially 571:579 partially 571:579 Both partially and non-glycosylated CLN3 were transported correctly to lysosomes. 17286803 6 27 theme Mevalonate 648:657 arg1 incorporation 659:671 Mevalonate incorporation 648:671 Mevalonate incorporation 648:671 Mevalonate incorporation and farnesyltransferase inhibitor studies indicate that CLN3 is prenylated most likely at cysteine 435. 17286803 3 28 theme glycosylation 351:363 arg1 role 343:346 the role 339:346 the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells 339:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 10 29 theme efficient 1511:1519 arg1 sorting 1521:1527 efficient sorting 1511:1527 efficient sorting 1511:1527 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 3 30 theme non-neuronal 429:440 arg1 cells 455:459 non-neuronal and neuronal cells 429:459 cells 455:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 0 31 theme C-terminal 0:9 arg1 prenylation 11:21 C-terminal prenylation 0:21 C-terminal prenylation of the CLN3 membrane glycoprotein 0:55 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 1 32 from Mutations 115:123 arg1 CLN3 174:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 9 33 theme length 1338:1343 arg1 composition 1349:1359 composition 1349:1359 composition 1349:1359 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 33 theme length 1338:1343 arg1 domain 1321:1326 the C-terminal domain 1306:1326 the C-terminal domain of proper length and composition 1306:1359 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 33 theme length 1338:1343 arg1 length 1338:1343 proper length 1331:1343 proper length 1331:1343 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 1 34 theme severe 191:196 arg1 disorder 216:223 a severe neurodegenerative disorder 189:223 a severe neurodegenerative disorder 189:223 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 3 35 theme lysosomal 398:406 arg1 transport 408:416 lysosomal transport 398:416 lysosomal transport of CLN3 in non-neuronal and neuronal cells 398:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 7 36 theme early 901:905 arg1 structures 917:926 early endosomal structures 901:926 early endosomal structures 901:926 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 1 37 theme neurodegenerative 198:214 arg1 disorder 216:223 a severe neurodegenerative disorder 189:223 a severe neurodegenerative disorder 189:223 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 10 38 theme different 1456:1464 arg1 CLN3 1489:1492 CLN3 1489:1492 CLN3 1489:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 10 38 theme different 1456:1464 arg1 domains 1478:1484 different cytoplasmic domains 1456:1484 different cytoplasmic domains of CLN3 1456:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 5 39 gly non-glycosylated 585:600 arg1 CLN3 602:605 Both partially and non-glycosylated CLN3 566:605 Both partially and non-glycosylated CLN3 566:605 Both partially and non-glycosylated CLN3 were transported correctly to lysosomes. 17286803 9 40 theme CLN3 1209:1212 arg1 CLN3 1209:1212 CLN3 1209:1212 CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1209:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 40 theme CLN3 1209:1212 arg1 domain 1199:1204 the entire C-terminal domain 1177:1204 the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1177:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 41 theme C-terminal 1310:1319 arg1 composition 1349:1359 composition 1349:1359 composition 1349:1359 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 41 theme C-terminal 1310:1319 arg1 domain 1321:1326 the C-terminal domain 1306:1326 the C-terminal domain of proper length and composition 1306:1359 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 41 theme C-terminal 1310:1319 arg1 length 1338:1343 proper length 1331:1343 proper length 1331:1343 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 7 42 theme impaired 881:888 arg1 sorting 890:896 impaired sorting 881:896 impaired sorting in early endosomal structures 881:926 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 10 43 from structures 1442:1451 arg1 CLN3 1489:1492 CLN3 1489:1492 CLN3 1489:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 10 43 from structures 1442:1451 arg1 domains 1478:1484 different cytoplasmic domains 1456:1484 different cytoplasmic domains of CLN3 1456:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 0 44 theme membrane 35:42 arg1 glycoprotein 44:55 the CLN3 membrane glycoprotein 26:55 the CLN3 membrane glycoprotein 26:55 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 9 45 theme domain 1321:1326 arg1 importance 1292:1301 the importance 1288:1301 the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum 1288:1397 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 2 46 theme cytosolic 258:266 arg1 structures 275:284 two cytosolic signal structures 254:284 two cytosolic signal structures contributing to lysosomal targeting 254:320 Previous studies identified two cytosolic signal structures contributing to lysosomal targeting. 17286803 9 47 theme C-terminal 1188:1197 arg1 CLN3 1209:1212 CLN3 1209:1212 CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1209:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 47 theme C-terminal 1188:1197 arg1 domain 1199:1204 the entire C-terminal domain 1177:1204 the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1177:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 0 48 theme CLN3 30:33 arg1 glycoprotein 44:55 the CLN3 membrane glycoprotein 26:55 the CLN3 membrane glycoprotein 26:55 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 3 49 theme neuronal 446:453 arg1 cells 455:459 non-neuronal and neuronal cells 429:459 cells 455:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 4 50 theme COS7 499:502 arg1 cells 504:508 COS7 cells 499:508 COS7 cells 499:508 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 7 51 from sorting 890:896 arg1 structures 917:926 early endosomal structures 901:926 early endosomal structures 901:926 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 9 52 theme domain 1199:1204 arg1 Alteration 1076:1085 Alteration 1076:1085 Alteration of the spacing between the transmembrane domain and the CAAX motif 1076:1152 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 52 theme domain 1199:1204 arg1 substitution 1161:1172 the substitution 1157:1172 the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1157:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 53 theme proper 1331:1336 arg1 length 1338:1343 proper length 1331:1343 proper length 1331:1343 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 8 54 theme surface 984:990 arg1 expression 992:1001 the cell surface expression 975:1001 the cell surface expression of CLN3 975:1009 Additionally, the cell surface expression of CLN3 was increased in the presence of farnesyltransferase inhibitors. 17286803 0 55 theme glycoprotein 44:55 arg1 prenylation 11:21 C-terminal prenylation 0:21 C-terminal prenylation of the CLN3 membrane glycoprotein 0:55 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 10 56 theme CLN3 1489:1492 arg1 CLN3 1489:1492 CLN3 1489:1492 CLN3 1489:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 10 56 theme CLN3 1489:1492 arg1 domains 1478:1484 different cytoplasmic domains 1456:1484 different cytoplasmic domains of CLN3 1456:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 2 57 theme Previous 226:233 arg1 studies 235:241 Previous studies 226:241 Previous studies 226:241 Previous studies identified two cytosolic signal structures contributing to lysosomal targeting. 17286803 8 58 theme CLN3 1006:1009 arg1 expression 992:1001 the cell surface expression 975:1001 the cell surface expression of CLN3 975:1009 Additionally, the cell surface expression of CLN3 was increased in the presence of farnesyltransferase inhibitors. 17286803 9 59 theme spacing 1094:1100 arg1 Alteration 1076:1085 Alteration 1076:1085 Alteration of the spacing between the transmembrane domain and the CAAX motif 1076:1152 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 59 theme spacing 1094:1100 arg1 substitution 1161:1172 the substitution 1157:1172 the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1157:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 7 60 theme CLN3 840:843 arg1 level 831:835 the steady-state level 814:835 the steady-state level of CLN3 in lysosomes most likely 814:868 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 3 61 gly glycosylation 351:363 arg1 CLN3 421:424 CLN3 421:424 CLN3 421:424 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 17286803 9 62 theme receptors 1260:1268 arg1 tails 1231:1235 cytoplasmic tails 1219:1235 cytoplasmic tails of mannose 6-phosphate receptors 1219:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 7 63 theme steady-state 818:829 arg1 level 831:835 the steady-state level 814:835 the steady-state level of CLN3 in lysosomes most likely 814:868 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 1 64 gly glycoprotein 161:172 arg1 glycoprotein 161:172 the polytopic lysosomal membrane glycoprotein CLN3 128:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 9 65 theme composition 1349:1359 arg1 composition 1349:1359 composition 1349:1359 composition 1349:1359 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 65 theme composition 1349:1359 arg1 domain 1321:1326 the C-terminal domain 1306:1326 the C-terminal domain of proper length and composition 1306:1359 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 65 theme composition 1349:1359 arg1 length 1338:1343 proper length 1331:1343 proper length 1331:1343 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 10 66 theme signal 1435:1440 arg1 structures 1442:1451 co-operative signal structures 1422:1451 co-operative signal structures in different cytoplasmic domains of CLN3 1422:1492 The data suggest that co-operative signal structures in different cytoplasmic domains of CLN3 are required for efficient sorting and for transport to the lysosome. 17286803 9 67 theme cytoplasmic 1219:1229 arg1 tails 1231:1235 cytoplasmic tails 1219:1235 cytoplasmic tails of mannose 6-phosphate receptors 1219:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 1 68 theme polytopic 132:140 arg1 CLN3 174:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 7 69 theme neuronal 945:952 arg1 cells 954:958 neuronal cells 945:958 neuronal cells 945:958 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 7 70 theme endosomal 907:915 arg1 structures 917:926 early endosomal structures 901:926 early endosomal structures 901:926 Substitution of cysteine 435 reduced the steady-state level of CLN3 in lysosomes most likely because of impaired sorting in early endosomal structures, particularly in neuronal cells. 17286803 4 71 theme Mutational 462:471 arg1 analysis 473:480 Mutational analysis 462:480 Mutational analysis 462:480 Mutational analysis revealed that in COS7 cells, CLN3 is glycosylated at asparagine residues 71 and 85. 17286803 0 72 gly glycoprotein 44:55 arg1 glycoprotein 44:55 the CLN3 membrane glycoprotein 26:55 the CLN3 membrane glycoprotein 26:55 C-terminal prenylation of the CLN3 membrane glycoprotein is required for efficient endosomal sorting to lysosomes. 17286803 9 73 theme entire 1181:1186 arg1 CLN3 1209:1212 CLN3 1209:1212 CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1209:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 73 theme entire 1181:1186 arg1 domain 1199:1204 the entire C-terminal domain 1177:1204 the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors 1177:1268 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 1 74 theme lysosomal 142:150 arg1 CLN3 174:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 9 75 theme transmembrane 1114:1126 arg1 domain 1128:1133 the transmembrane domain 1110:1133 the transmembrane domain 1110:1133 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 9 76 theme CAAX 1143:1146 arg1 motif 1148:1152 the CAAX motif 1139:1152 the CAAX motif 1139:1152 Alteration of the spacing between the transmembrane domain and the CAAX motif or the substitution of the entire C-terminal domain of CLN3 with cytoplasmic tails of mannose 6-phosphate receptors have demonstrated the importance of the C-terminal domain of proper length and composition for exit of the endoplasmic reticulum. 17286803 2 77 theme signal 268:273 arg1 structures 275:284 two cytosolic signal structures 254:284 two cytosolic signal structures contributing to lysosomal targeting 254:320 Previous studies identified two cytosolic signal structures contributing to lysosomal targeting. 17286803 1 78 theme membrane 152:159 arg1 CLN3 174:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 the polytopic lysosomal membrane glycoprotein CLN3 128:177 Mutations in the polytopic lysosomal membrane glycoprotein CLN3 result in a severe neurodegenerative disorder. 17286803 6 79 theme inhibitor 697:705 arg1 studies 707:713 Mevalonate incorporation and farnesyltransferase inhibitor studies 648:713 Mevalonate incorporation and farnesyltransferase inhibitor studies 648:713 Mevalonate incorporation and farnesyltransferase inhibitor studies indicate that CLN3 is prenylated most likely at cysteine 435. 17286803 3 80 from role 343:346 arg1 transport 408:416 lysosomal transport 398:416 lysosomal transport of CLN3 in non-neuronal and neuronal cells 398:459 We now examined the role of glycosylation and the C-terminal CAAX motif in lysosomal transport of CLN3 in non-neuronal and neuronal cells. 18768590 0 0 theme kinase-dependent 76:91 arg1 pathway 93:99 a kinase-dependent pathway 74:99 a kinase-dependent pathway 74:99 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 1 1 theme K+ 302:303 arg1 channel 305:311 the renal outer medullary K+ channel 276:311 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 1 theme K+ 302:303 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 2 2 theme laevis 519:524 arg1 oocytes 526:532 Xenopus laevis oocytes 511:532 Xenopus laevis oocytes 511:532 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 9 3 theme TRPV5 1501:1505 arg1 delivery 1482:1489 the enhanced delivery 1469:1489 the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway 1469:1582 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 1 4 theme Lysine 132:137 arg1 family 143:148 No Lysine (K) family 129:148 No Lysine (K) family of protein serine/threonine kinases 129:184 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 5 dep cotransporters 257:270 arg1 channel 305:311 the renal outer medullary K+ channel 276:311 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 5 dep cotransporters 257:270 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 5 dep cotransporters 257:270 arg1 channel 328:334 Cl(-) channel 322:334 Cl(-) channel SLC26A9 322:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 2 6 theme Xenopus 511:517 arg1 oocytes 526:532 Xenopus laevis oocytes 511:532 Xenopus laevis oocytes 511:532 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 7 theme TRPV5 569:573 arg1 expression 555:564 expression 555:564 expression 555:564 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 7 theme TRPV5 569:573 arg1 function 542:549 function 542:549 function 542:549 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 7 8 theme glycosylated 1189:1200 arg1 TRPV5 1202:1206 The complexly glycosylated TRPV5 1175:1206 The complexly glycosylated TRPV5 that appears at the plasma membrane 1175:1242 The complexly glycosylated TRPV5 that appears at the plasma membrane was increased by WNK3. 18768590 9 9 theme plasma 1514:1519 arg1 membrane 1521:1528 the plasma membrane 1510:1528 the plasma membrane from its intracellular pool via the secretory pathway 1510:1582 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 5 10 theme Ca2+ 931:934 arg1 absorption 936:945 active intestinal Ca2+ absorption 913:945 active intestinal Ca2+ absorption 913:945 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 7 11 theme plasma 1228:1233 arg1 membrane 1235:1242 the plasma membrane 1224:1242 the plasma membrane 1224:1242 The complexly glycosylated TRPV5 that appears at the plasma membrane was increased by WNK3. 18768590 6 12 from mutation 1116:1123 arg1 WNK3 1128:1131 WNK3 1128:1131 WNK3 1128:1131 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 9 13 theme increased 1405:1413 arg1 due 1462:1464 due 1462:1464 due 1462:1464 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 9 13 theme increased 1405:1413 arg1 expression 1431:1440 The increased plasma membrane expression 1401:1440 The increased plasma membrane expression of TRPV5 1401:1449 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 5 14 theme Ca2+ 814:817 arg1 influx 819:824 Ca2+ influx 814:824 Ca2+ influx 814:824 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 1 15 theme SLC12A 224:229 arg1 family 231:236 the SLC12A family 220:236 the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 220:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 16 theme Cl 322:323 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 16 theme Cl 322:323 arg1 channel 328:334 Cl(-) channel 322:334 Cl(-) channel SLC26A9 322:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 6 17 theme kinase 952:957 arg1 WNK3 969:972 WNK3 969:972 WNK3 alone 969:978 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 6 17 theme kinase 952:957 arg1 sufficient 984:993 sufficient 984:993 sufficient 984:993 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 6 17 theme kinase 952:957 arg1 domain 959:964 The kinase domain 948:964 The kinase domain of WNK3 alone 948:978 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 5 18 theme closest 874:880 arg1 TRPV6 854:858 TRPV6 854:858 TRPV6 854:858 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 5 18 theme closest 874:880 arg1 homolog 882:888 the closest homolog 870:888 the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption 870:945 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 9 19 theme secretory 1566:1574 arg1 pathway 1576:1582 the secretory pathway 1562:1582 the secretory pathway 1562:1582 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 10 20 theme transcellular 1649:1661 arg1 pathway 1678:1684 the transcellular Ca2+ transport pathway 1645:1684 the transcellular Ca2+ transport pathway 1645:1684 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 2 21 theme Ca2+ 405:408 arg1 TRPV5 379:383 TRPV5 379:383 TRPV5 379:383 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 21 theme Ca2+ 405:408 arg1 gatekeeper 435:444 a gatekeeper 433:444 a gatekeeper for active Ca2+ reabsorption 433:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 21 theme Ca2+ 405:408 arg1 channel 410:416 a renal epithelial Ca2+ channel 386:416 a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption 386:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 9 22 theme intracellular 1539:1551 arg1 pool 1553:1556 its intracellular pool 1535:1556 its intracellular pool 1535:1556 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 1 23 dep channel 305:311 arg1 ROMK 313:316 ROMK 313:316 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 8 24 theme TRPV5 1285:1289 arg1 exocytosis 1271:1280 The exocytosis 1267:1280 The exocytosis of TRPV5 1267:1289 The exocytosis of TRPV5 was increased by WNK3, and the effect of WNK3 on TRPV5 was abolished by the microtubule inhibitor colchicine. 18768590 3 25 theme TRPV5-mediated 632:645 arg1 uptake 652:657 TRPV5-mediated Ca2+ uptake 632:657 TRPV5-mediated Ca2+ uptake 632:657 An 82.7 +/- 7.1% increase in TRPV5-mediated Ca2+ uptake was observed when WNK3 was coexpressed. 18768590 6 26 theme WNK3 969:972 arg1 WNK3 969:972 WNK3 969:972 WNK3 alone 969:978 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 6 26 theme WNK3 969:972 arg1 sufficient 984:993 sufficient 984:993 sufficient 984:993 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 6 26 theme WNK3 969:972 arg1 domain 959:964 The kinase domain 948:964 The kinase domain of WNK3 alone 948:978 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 3 27 from increase 620:627 arg1 uptake 652:657 TRPV5-mediated Ca2+ uptake 632:657 TRPV5-mediated Ca2+ uptake 632:657 An 82.7 +/- 7.1% increase in TRPV5-mediated Ca2+ uptake was observed when WNK3 was coexpressed. 18768590 1 28 theme family 231:236 arg1 members 209:215 members 209:215 members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 209:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 6 29 theme kinase-dependent 1147:1162 arg1 mechanism 1164:1172 a kinase-dependent mechanism 1145:1172 a kinase-dependent mechanism 1145:1172 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 6 30 theme regulatory 1055:1064 arg1 effect 1066:1071 the positive regulatory effect 1042:1071 the positive regulatory effect 1042:1071 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 4 31 theme voltage-clamp 770:782 arg1 technique 784:792 the voltage-clamp technique 766:792 the voltage-clamp technique 766:792 A similar increase in TRPV5-mediated Na+ current was observed with the voltage-clamp technique. 18768590 0 32 theme epithelial 26:35 arg1 channels 45:52 epithelial calcium channels TRPV5 and TRPV6 26:68 epithelial calcium channels TRPV5 and TRPV6 26:68 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 0 32 theme epithelial 26:35 arg1 TRPV6 64:68 TRPV6 64:68 TRPV6 64:68 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 0 32 theme epithelial 26:35 arg1 TRPV5 54:58 TRPV5 54:58 TRPV5 54:58 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 8 33 theme WNK3 1332:1335 arg1 effect 1322:1327 the effect 1318:1327 the effect of WNK3 on TRPV5 1318:1344 The exocytosis of TRPV5 was increased by WNK3, and the effect of WNK3 on TRPV5 was abolished by the microtubule inhibitor colchicine. 18768590 9 34 from pool 1553:1556 arg1 membrane 1521:1528 the plasma membrane 1510:1528 the plasma membrane from its intracellular pool via the secretory pathway 1510:1582 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 6 35 theme positive 1046:1053 arg1 effect 1066:1071 the positive regulatory effect 1042:1071 the positive regulatory effect 1042:1071 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 5 36 theme active 913:918 arg1 absorption 936:945 active intestinal Ca2+ absorption 913:945 active intestinal Ca2+ absorption 913:945 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 1 37 theme protein 153:159 arg1 kinases 178:184 protein serine/threonine kinases 153:184 protein serine/threonine kinases 153:184 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 6 38 theme Ca2+ 1022:1025 arg1 transport 1027:1035 TRPV5-mediated Ca2+ transport 1007:1035 TRPV5-mediated Ca2+ transport 1007:1035 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 1 39 theme cation-chloride 241:255 arg1 channel 305:311 the renal outer medullary K+ channel 276:311 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 39 theme cation-chloride 241:255 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 39 theme cation-chloride 241:255 arg1 channel 328:334 Cl(-) channel 322:334 Cl(-) channel SLC26A9 322:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 10 40 theme positive 1623:1630 arg1 regulator 1632:1640 a positive regulator 1621:1640 a positive regulator of the transcellular Ca2+ transport pathway 1621:1684 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 10 40 theme positive 1623:1630 arg1 WNK3 1613:1616 WNK3 1613:1616 WNK3 1613:1616 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 10 41 theme Ca2+ 1663:1666 arg1 pathway 1678:1684 the transcellular Ca2+ transport pathway 1645:1684 the transcellular Ca2+ transport pathway 1645:1684 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 8 42 from effect 1322:1327 arg1 TRPV5 1340:1344 TRPV5 1340:1344 TRPV5 1340:1344 The exocytosis of TRPV5 was increased by WNK3, and the effect of WNK3 on TRPV5 was abolished by the microtubule inhibitor colchicine. 18768590 1 43 theme serine/threonine 161:176 arg1 kinases 178:184 protein serine/threonine kinases 153:184 protein serine/threonine kinases 153:184 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 2 44 theme epithelial 394:403 arg1 TRPV5 379:383 TRPV5 379:383 TRPV5 379:383 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 44 theme epithelial 394:403 arg1 gatekeeper 435:444 a gatekeeper 433:444 a gatekeeper for active Ca2+ reabsorption 433:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 44 theme epithelial 394:403 arg1 channel 410:416 a renal epithelial Ca2+ channel 386:416 a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption 386:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 1 45 theme cotransporters 257:270 arg1 family 231:236 the SLC12A family 220:236 the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 220:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 2 46 theme Ca2+ 457:460 arg1 reabsorption 462:473 active Ca2+ reabsorption 450:473 active Ca2+ reabsorption 450:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 4 47 theme TRPV5-mediated 721:734 arg1 current 740:746 TRPV5-mediated Na+ current 721:746 TRPV5-mediated Na+ current 721:746 A similar increase in TRPV5-mediated Na+ current was observed with the voltage-clamp technique. 18768590 6 48 theme kinase-inactive 1094:1108 arg1 mutation 1116:1123 the kinase-inactive D294A mutation 1090:1123 the kinase-inactive D294A mutation in WNK3 1090:1131 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 1 49 theme kinases 178:184 arg1 family 143:148 No Lysine (K) family 129:148 No Lysine (K) family of protein serine/threonine kinases 129:184 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 0 50 theme calcium 37:43 arg1 channels 45:52 epithelial calcium channels TRPV5 and TRPV6 26:68 epithelial calcium channels TRPV5 and TRPV6 26:68 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 0 50 theme calcium 37:43 arg1 TRPV6 64:68 TRPV6 64:68 TRPV6 64:68 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 0 50 theme calcium 37:43 arg1 TRPV5 54:58 TRPV5 54:58 TRPV5 54:58 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 3 51 dep % 618:618 arg1 7.1 615:617 7.1 615:617 7.1 615:617 An 82.7 +/- 7.1% increase in TRPV5-mediated Ca2+ uptake was observed when WNK3 was coexpressed. 18768590 3 51 dep % 618:618 arg1 82.7 606:609 82.7 606:609 82.7 606:609 An 82.7 +/- 7.1% increase in TRPV5-mediated Ca2+ uptake was observed when WNK3 was coexpressed. 18768590 2 52 theme renal 388:392 arg1 TRPV5 379:383 TRPV5 379:383 TRPV5 379:383 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 52 theme renal 388:392 arg1 gatekeeper 435:444 a gatekeeper 433:444 a gatekeeper for active Ca2+ reabsorption 433:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 52 theme renal 388:392 arg1 channel 410:416 a renal epithelial Ca2+ channel 386:416 a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption 386:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 53 theme WNK3 371:374 arg1 effect 361:366 the effect 357:366 the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption 357:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 10 54 theme pathway 1678:1684 arg1 regulator 1632:1640 a positive regulator 1621:1640 a positive regulator of the transcellular Ca2+ transport pathway 1621:1684 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 10 54 theme pathway 1678:1684 arg1 WNK3 1613:1616 WNK3 1613:1616 WNK3 1613:1616 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 3 55 dep increase 620:627 arg1 % 618:618 % 618:618 % 618:618 An 82.7 +/- 7.1% increase in TRPV5-mediated Ca2+ uptake was observed when WNK3 was coexpressed. 18768590 9 56 theme mature 1494:1499 arg1 TRPV5 1501:1505 mature TRPV5 1494:1505 mature TRPV5 1494:1505 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 0 57 dep channels 45:52 arg1 channels 45:52 epithelial calcium channels TRPV5 and TRPV6 26:68 epithelial calcium channels TRPV5 and TRPV6 26:68 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 0 57 dep channels 45:52 arg1 TRPV6 64:68 TRPV6 64:68 TRPV6 64:68 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 0 57 dep channels 45:52 arg1 TRPV5 54:58 TRPV5 54:58 TRPV5 54:58 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. 18768590 2 58 theme active 450:455 arg1 reabsorption 462:473 active Ca2+ reabsorption 450:473 active Ca2+ reabsorption 450:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 1 59 theme renal 280:284 arg1 channel 305:311 the renal outer medullary K+ channel 276:311 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 59 theme renal 280:284 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 9 60 theme TRPV5 1445:1449 arg1 due 1462:1464 due 1462:1464 due 1462:1464 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 9 60 theme TRPV5 1445:1449 arg1 expression 1431:1440 The increased plasma membrane expression 1401:1440 The increased plasma membrane expression of TRPV5 1401:1449 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 4 61 theme Na+ 736:738 arg1 current 740:746 TRPV5-mediated Na+ current 721:746 TRPV5-mediated Na+ current 721:746 A similar increase in TRPV5-mediated Na+ current was observed with the voltage-clamp technique. 18768590 5 62 theme Na+ 830:832 arg1 current 834:840 Na+ current 830:840 Na+ current 830:840 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 1 63 theme outer 286:290 arg1 channel 305:311 the renal outer medullary K+ channel 276:311 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 63 theme outer 286:290 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 7 64 gly glycosylated 1189:1200 arg1 TRPV5 1202:1206 The complexly glycosylated TRPV5 1175:1206 The complexly glycosylated TRPV5 that appears at the plasma membrane 1175:1242 The complexly glycosylated TRPV5 that appears at the plasma membrane was increased by WNK3. 18768590 5 65 theme intestinal 920:929 arg1 absorption 936:945 active intestinal Ca2+ absorption 913:945 active intestinal Ca2+ absorption 913:945 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 3 66 theme Ca2+ 647:650 arg1 uptake 652:657 TRPV5-mediated Ca2+ uptake 632:657 TRPV5-mediated Ca2+ uptake 632:657 An 82.7 +/- 7.1% increase in TRPV5-mediated Ca2+ uptake was observed when WNK3 was coexpressed. 18768590 9 67 theme enhanced 1473:1480 arg1 delivery 1482:1489 the enhanced delivery 1469:1489 the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway 1469:1582 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 8 68 theme inhibitor 1379:1387 arg1 colchicine 1389:1398 the microtubule inhibitor colchicine 1363:1398 the microtubule inhibitor colchicine 1363:1398 The exocytosis of TRPV5 was increased by WNK3, and the effect of WNK3 on TRPV5 was abolished by the microtubule inhibitor colchicine. 18768590 5 69 theme TRPV5 893:897 arg1 TRPV6 854:858 TRPV6 854:858 TRPV6 854:858 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 5 69 theme TRPV5 893:897 arg1 homolog 882:888 the closest homolog 870:888 the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption 870:945 WNK3 also enhanced Ca2+ influx and Na+ current mediated by TRPV6, which is the closest homolog of TRPV5 that mediates active intestinal Ca2+ absorption. 18768590 4 70 from increase 709:716 arg1 current 740:746 TRPV5-mediated Na+ current 721:746 TRPV5-mediated Na+ current 721:746 A similar increase in TRPV5-mediated Na+ current was observed with the voltage-clamp technique. 18768590 1 71 theme the 120:122 arg1 WNK3 102:105 WNK3 102:105 WNK3 102:105 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 71 theme the 120:122 arg1 member 110:115 a member 108:115 a member of the With No Lysine (K) family of protein serine/threonine kinases 108:184 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 10 72 theme transport 1668:1676 arg1 pathway 1678:1684 the transcellular Ca2+ transport pathway 1645:1684 the transcellular Ca2+ transport pathway 1645:1684 These results indicate that WNK3 is a positive regulator of the transcellular Ca2+ transport pathway. 18768590 8 73 theme microtubule 1367:1377 arg1 colchicine 1389:1398 the microtubule inhibitor colchicine 1363:1398 the microtubule inhibitor colchicine 1363:1398 The exocytosis of TRPV5 was increased by WNK3, and the effect of WNK3 on TRPV5 was abolished by the microtubule inhibitor colchicine. 18768590 6 74 theme D294A 1110:1114 arg1 mutation 1116:1123 the kinase-inactive D294A mutation 1090:1123 the kinase-inactive D294A mutation in WNK3 1090:1131 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 9 75 theme plasma 1415:1420 arg1 due 1462:1464 due 1462:1464 due 1462:1464 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 9 75 theme plasma 1415:1420 arg1 expression 1431:1440 The increased plasma membrane expression 1401:1440 The increased plasma membrane expression of TRPV5 1401:1449 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 6 76 theme TRPV5-mediated 1007:1020 arg1 transport 1027:1035 TRPV5-mediated Ca2+ transport 1007:1035 TRPV5-mediated Ca2+ transport 1007:1035 The kinase domain of WNK3 alone was sufficient to increase TRPV5-mediated Ca2+ transport, and the positive regulatory effect was abolished by the kinase-inactive D294A mutation in WNK3, indicating a kinase-dependent mechanism. 18768590 2 77 dep function 542:549 arg1 the 538:540 the 538:540 the 538:540 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 1 78 theme medullary 292:300 arg1 channel 305:311 the renal outer medullary K+ channel 276:311 the renal outer medullary K+ channel ROMK 276:316 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 1 78 theme medullary 292:300 arg1 cotransporters 257:270 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9 241:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 18768590 4 79 theme similar 701:707 arg1 increase 709:716 A similar increase 699:716 A similar increase in TRPV5-mediated Na+ current 699:746 A similar increase in TRPV5-mediated Na+ current was observed with the voltage-clamp technique. 18768590 2 80 from effect 361:366 arg1 TRPV5 379:383 TRPV5 379:383 TRPV5 379:383 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 2 80 from effect 361:366 arg1 channel 410:416 a renal epithelial Ca2+ channel 386:416 a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption 386:473 To evaluate the effect of WNK3 on TRPV5, a renal epithelial Ca2+ channel that serves as a gatekeeper for active Ca2+ reabsorption, WNK3 and TRPV5 were coexpressed in Xenopus laevis oocytes and the function and expression of TRPV5 were subsequently examined. 18768590 9 81 theme membrane 1422:1429 arg1 due 1462:1464 due 1462:1464 due 1462:1464 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 9 81 theme membrane 1422:1429 arg1 expression 1431:1440 The increased plasma membrane expression 1401:1440 The increased plasma membrane expression of TRPV5 1401:1449 The increased plasma membrane expression of TRPV5 was likely due to the enhanced delivery of mature TRPV5 to the plasma membrane from its intracellular pool via the secretory pathway. 18768590 1 82 dep channel 328:334 arg1 SLC26A9 336:342 SLC26A9 336:342 Cl(-) channel SLC26A9 322:342 WNK3, a member of the With No Lysine (K) family of protein serine/threonine kinases, was shown to regulate members of the SLC12A family of cation-chloride cotransporters and the renal outer medullary K+ channel ROMK and Cl(-) channel SLC26A9. 2148568 0 0 theme lung 102:105 arg1 fibroblasts 107:117 human lung fibroblasts 96:117 human lung fibroblasts 96:117 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 2 1 theme cell-associated 386:400 arg1 forms 402:406 the hydrophobic cell-associated forms 370:406 the hydrophobic cell-associated forms 370:406 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 2 1 theme cell-associated 386:400 arg1 precursors 415:424 precursors 415:424 precursors for the nonhydrophobic medium-released species 415:471 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 0 2 theme human 96:100 arg1 fibroblasts 107:117 human lung fibroblasts 96:117 human lung fibroblasts 96:117 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 5 3 theme lung 1001:1004 arg1 fibroblasts 1006:1016 human lung fibroblasts 995:1016 human lung fibroblasts 995:1016 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 2 4 theme hydrophobic 374:384 arg1 forms 402:406 the hydrophobic cell-associated forms 370:406 the hydrophobic cell-associated forms 370:406 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 2 4 theme hydrophobic 374:384 arg1 precursors 415:424 precursors 415:424 precursors for the nonhydrophobic medium-released species 415:471 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 3 5 theme medium-released 499:513 arg1 proteoglycans 515:527 the medium-released proteoglycans 495:527 the medium-released proteoglycans 495:527 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 5 6 theme phospholipid 1105:1116 arg1 anchor 1118:1123 the phospholipid anchor 1101:1123 the phospholipid anchor 1101:1123 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 3 7 theme hydrophobic 568:578 arg1 species 596:602 the hydrophobic cell-associated species 564:602 the hydrophobic cell-associated species 564:602 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 5 8 theme human 995:999 arg1 fibroblasts 1006:1016 human lung fibroblasts 995:1016 human lung fibroblasts 995:1016 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 4 9 theme phosphatidylinositol-specific 804:832 arg1 C 848:848 bacterial phosphatidylinositol-specific phospholipase C 794:848 bacterial phosphatidylinositol-specific phospholipase C 794:848 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 3 10 theme acid 632:635 arg1 identical 665:673 identical 665:673 identical 665:673 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 10 theme acid 632:635 arg1 sequences 637:645 the NH2-terminal amino acid sequences 609:645 the NH2-terminal amino acid sequences of both forms 609:659 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 4 11 theme C 848:848 arg1 effect 784:789 the effect 780:789 the effect of bacterial phosphatidylinositol-specific phospholipase C 780:848 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 11 theme C 848:848 arg1 characterization 680:695 The characterization 676:695 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins 676:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 12 theme core 762:765 arg1 proteins 767:774 these core proteins 756:774 these core proteins 756:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 13 theme fibroblast 711:720 arg1 message 744:750 the message 740:750 the message for these core proteins 740:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 13 theme fibroblast 711:720 arg1 cDNAs 722:726 human lung fibroblast cDNAs 700:726 human lung fibroblast cDNAs that encode the message for these core proteins 700:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 14 theme bacterial 794:802 arg1 C 848:848 bacterial phosphatidylinositol-specific phospholipase C 794:848 bacterial phosphatidylinositol-specific phospholipase C 794:848 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 5 15 theme membrane 970:977 arg1 proteoglycan 979:990 a novel membrane proteoglycan 962:990 a novel membrane proteoglycan in human lung fibroblasts 962:1016 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 1 16 theme membrane 172:179 arg1 proteoglycan 197:208 a membrane heparan sulfate proteoglycan 170:208 a membrane heparan sulfate proteoglycan 170:208 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 1 17 theme heparan 181:187 arg1 proteoglycan 197:208 a membrane heparan sulfate proteoglycan 170:208 a membrane heparan sulfate proteoglycan 170:208 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 4 18 theme lung 706:709 arg1 message 744:750 the message 740:750 the message for these core proteins 740:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 18 theme lung 706:709 arg1 cDNAs 722:726 human lung fibroblast cDNAs 700:726 human lung fibroblast cDNAs that encode the message for these core proteins 700:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 2 19 theme Pulse-chase 337:347 arg1 studies 349:355 Pulse-chase studies 337:355 Pulse-chase studies 337:355 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 1 20 theme sulfate 189:195 arg1 proteoglycan 197:208 a membrane heparan sulfate proteoglycan 170:208 a membrane heparan sulfate proteoglycan 170:208 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 0 21 theme Molecular 0:8 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 0:118 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 5 22 theme novel 964:968 arg1 proteoglycan 979:990 a novel membrane proteoglycan 962:990 a novel membrane proteoglycan in human lung fibroblasts 962:1016 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 1 23 theme proteoglycan 197:208 arg1 protein 159:165 the 64-kD core protein 144:165 the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts 144:236 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 1 24 theme culture 308:314 arg1 medium 316:321 the culture medium 304:321 the culture medium of the cells 304:334 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 3 25 theme NH2-terminal 613:624 arg1 identical 665:673 identical 665:673 identical 665:673 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 25 theme NH2-terminal 613:624 arg1 sequences 637:645 the NH2-terminal amino acid sequences 609:645 the NH2-terminal amino acid sequences of both forms 609:659 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 5 26 theme proteoglycan 1054:1065 arg1 shedding 1037:1044 the shedding 1033:1044 the shedding of this proteoglycan 1033:1065 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 5 26 theme proteoglycan 1054:1065 arg1 related 1074:1080 related 1074:1080 related 1074:1080 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 3 27 theme proteoglycans 515:527 arg1 proteins 483:490 The core proteins 474:490 The core proteins of the medium-released proteoglycans 474:527 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 27 theme proteoglycans 515:527 arg1 smaller 542:548 smaller 542:548 smaller 542:548 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 5 28 from proteoglycan 979:990 arg1 fibroblasts 1006:1016 human lung fibroblasts 995:1016 human lung fibroblasts 995:1016 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 3 29 theme amino 626:630 arg1 identical 665:673 identical 665:673 identical 665:673 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 29 theme amino 626:630 arg1 sequences 637:645 the NH2-terminal amino acid sequences 609:645 the NH2-terminal amino acid sequences of both forms 609:659 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 1 30 theme human 215:219 arg1 fibroblasts 226:236 human lung fibroblasts 215:236 human lung fibroblasts 215:236 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 0 31 theme membrane 53:60 arg1 proteoglycan 78:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 2 32 theme medium-released 449:463 arg1 species 465:471 the nonhydrophobic medium-released species 430:471 the nonhydrophobic medium-released species 430:471 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 3 33 theme forms 655:659 arg1 identical 665:673 identical 665:673 identical 665:673 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 33 theme forms 655:659 arg1 sequences 637:645 the NH2-terminal amino acid sequences 609:645 the NH2-terminal amino acid sequences of both forms 609:659 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 34 theme core 478:481 arg1 proteins 483:490 The core proteins 474:490 The core proteins of the medium-released proteoglycans 474:527 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 3 34 theme core 478:481 arg1 smaller 542:548 smaller 542:548 smaller 542:548 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 1 35 theme lung 221:224 arg1 fibroblasts 226:236 human lung fibroblasts 215:236 human lung fibroblasts 215:236 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 0 36 theme phosphatidylinositol-anchored 23:51 arg1 proteoglycan 78:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 5 37 theme anchor 1118:1123 arg1 presence 1089:1096 the presence 1085:1096 the presence of the phospholipid anchor 1085:1123 These data identify a novel membrane proteoglycan in human lung fibroblasts and imply that the shedding of this proteoglycan may be related to the presence of the phospholipid anchor. 2148568 2 38 theme nonhydrophobic 434:447 arg1 species 465:471 the nonhydrophobic medium-released species 430:471 the nonhydrophobic medium-released species 430:471 Pulse-chase studies suggest that the hydrophobic cell-associated forms act as precursors for the nonhydrophobic medium-released species. 2148568 0 39 theme sulfate 70:76 arg1 proteoglycan 78:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 1 40 theme cells 330:334 arg1 medium 316:321 the culture medium 304:321 the culture medium of the cells 304:334 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 1 41 from fibroblasts 226:236 arg1 protein 159:165 the 64-kD core protein 144:165 the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts 144:236 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 0 42 theme heparan 62:68 arg1 proteoglycan 78:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan 21:89 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 4 43 theme phospholipid 923:934 arg1 tail 936:939 a phospholipid tail 921:939 a phospholipid tail 921:939 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 44 theme phospholipase 834:846 arg1 C 848:848 bacterial phosphatidylinositol-specific phospholipase C 794:848 bacterial phosphatidylinositol-specific phospholipase C 794:848 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 45 theme hydrophobic 867:877 arg1 membrane-anchored 895:911 membrane-anchored 895:911 membrane-anchored 895:911 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 45 theme hydrophobic 867:877 arg1 proteoglycan 879:890 the hydrophobic proteoglycan 863:890 the hydrophobic proteoglycan 863:890 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 1 46 theme 64-kD 148:152 arg1 protein 159:165 the 64-kD core protein 144:165 the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts 144:236 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 0 47 from fibroblasts 107:117 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 0:118 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 4 48 theme human 700:704 arg1 message 744:750 the message 740:750 the message for these core proteins 740:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 48 theme human 700:704 arg1 cDNAs 722:726 human lung fibroblast cDNAs 700:726 human lung fibroblast cDNAs that encode the message for these core proteins 700:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 1 49 theme core 154:157 arg1 protein 159:165 the 64-kD core protein 144:165 the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts 144:236 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 2148568 4 50 theme cDNAs 722:726 arg1 effect 784:789 the effect 780:789 the effect of bacterial phosphatidylinositol-specific phospholipase C 780:848 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 4 50 theme cDNAs 722:726 arg1 characterization 680:695 The characterization 676:695 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins 676:774 The characterization of human lung fibroblast cDNAs that encode the message for these core proteins and the effect of bacterial phosphatidylinositol-specific phospholipase C suggest that the hydrophobic proteoglycan is membrane-anchored through a phospholipid tail. 2148568 0 51 theme proteoglycan 78:89 arg1 cloning 10:16 Molecular cloning 0:16 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 0:118 Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. 2148568 3 52 theme cell-associated 580:594 arg1 species 596:602 the hydrophobic cell-associated species 564:602 the hydrophobic cell-associated species 564:602 The core proteins of the medium-released proteoglycans are slightly smaller than those of the hydrophobic cell-associated species, but the NH2-terminal amino acid sequences of both forms are identical. 2148568 1 53 theme nonhydrophobic 255:268 arg1 proteoglycan 270:281 a nonhydrophobic proteoglycan 253:281 a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells 253:334 Two mAbs raised against the 64-kD core protein of a membrane heparan sulfate proteoglycan from human lung fibroblasts also recognize a nonhydrophobic proteoglycan which accumulates in the culture medium of the cells. 19651138 4 0 from contacts 484:491 arg1 form 523:526 a 4.1-A resolution crystal form 496:526 a 4.1-A resolution crystal form 496:526 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 4 1 theme tetrameric 537:546 arg1 arrangement 562:572 a tetrameric (dimer-dimer) arrangement 535:572 a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors 535:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 2 2 theme 1.8-A 296:300 arg1 structure 321:329 a 1.8-A resolution crystal structure 294:329 a 1.8-A resolution crystal structure of human GluR2-ATD 294:348 We report a 1.8-A resolution crystal structure of human GluR2-ATD. 19651138 4 3 theme 4.1-A 498:502 arg1 form 523:526 a 4.1-A resolution crystal form 496:526 a 4.1-A resolution crystal form 496:526 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 1 4 theme Ionotropic 141:150 arg1 receptors 162:170 Ionotropic glutamate receptors 141:170 Ionotropic glutamate receptors 141:170 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 3 5 theme dimeric 356:362 arg1 structure 364:372 This dimeric structure 351:372 This dimeric structure 351:372 This dimeric structure provides a mechanism for how the ATDs can drive receptor assembly and subtype-restricted composition. 19651138 3 6 theme subtype-restricted 444:461 arg1 composition 463:473 subtype-restricted composition 444:473 subtype-restricted composition 444:473 This dimeric structure provides a mechanism for how the ATDs can drive receptor assembly and subtype-restricted composition. 19651138 4 7 theme microscopic 626:636 arg1 data 638:641 previous cellular and cryo-electron microscopic data 590:641 previous cellular and cryo-electron microscopic data for full-length AMPA receptors 590:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 4 8 theme crystal 515:521 arg1 form 523:526 a 4.1-A resolution crystal form 496:526 a 4.1-A resolution crystal form 496:526 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 4 9 with consistent 574:583 arg1 data 638:641 previous cellular and cryo-electron microscopic data 590:641 previous cellular and cryo-electron microscopic data for full-length AMPA receptors 590:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 0 10 theme ionotropic 109:118 arg1 receptors 130:138 ionotropic glutamate receptors 109:138 ionotropic glutamate receptors 109:138 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 4 11 theme resolution 504:513 arg1 form 523:526 a 4.1-A resolution crystal form 496:526 a 4.1-A resolution crystal form 496:526 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 0 12 dep architecture 80:91 arg1 the 76:78 the 76:78 the 76:78 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 0 13 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the GluR2 amino-terminal domain 0:51 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 4 14 theme consistent 574:583 arg1 arrangement 562:572 a tetrameric (dimer-dimer) arrangement 535:572 a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors 535:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 0 15 theme receptors 130:138 arg1 assembly 97:104 assembly 97:104 assembly 97:104 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 0 15 theme receptors 130:138 arg1 architecture 80:91 architecture 80:91 architecture 80:91 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 2 16 theme crystal 313:319 arg1 structure 321:329 a 1.8-A resolution crystal structure 294:329 a 1.8-A resolution crystal structure of human GluR2-ATD 294:348 We report a 1.8-A resolution crystal structure of human GluR2-ATD. 19651138 1 17 theme common 208:213 arg1 architecture 215:226 a common architecture 206:226 a common architecture 206:226 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 4 18 theme cellular 599:606 arg1 data 638:641 previous cellular and cryo-electron microscopic data 590:641 previous cellular and cryo-electron microscopic data for full-length AMPA receptors 590:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 0 19 theme glutamate 120:128 arg1 receptors 130:138 ionotropic glutamate receptors 109:138 ionotropic glutamate receptors 109:138 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 4 20 theme Lattice 476:482 arg1 contacts 484:491 Lattice contacts 476:491 Lattice contacts in a 4.1-A resolution crystal form 476:526 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 2 21 theme resolution 302:311 arg1 structure 321:329 a 1.8-A resolution crystal structure 294:329 a 1.8-A resolution crystal structure of human GluR2-ATD 294:348 We report a 1.8-A resolution crystal structure of human GluR2-ATD. 19651138 0 22 theme amino-terminal 31:44 arg1 domain 46:51 the GluR2 amino-terminal domain 21:51 the GluR2 amino-terminal domain 21:51 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 4 23 theme previous 590:597 arg1 data 638:641 previous cellular and cryo-electron microscopic data 590:641 previous cellular and cryo-electron microscopic data for full-length AMPA receptors 590:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 1 24 contain have 201:204 arg1 receptors 162:170 Ionotropic glutamate receptors 141:170 Ionotropic glutamate receptors 141:170 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 1 24 contain have 201:204 arg2 architecture 215:226 a common architecture 206:226 a common architecture 206:226 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 0 25 theme GluR2 25:29 arg1 domain 46:51 the GluR2 amino-terminal domain 21:51 the GluR2 amino-terminal domain 21:51 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 4 26 theme dimer-dimer 549:559 arg1 arrangement 562:572 a tetrameric (dimer-dimer) arrangement 535:572 a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors 535:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 3 27 theme receptor 422:429 arg1 assembly 431:438 receptor assembly 422:438 receptor assembly 422:438 This dimeric structure provides a mechanism for how the ATDs can drive receptor assembly and subtype-restricted composition. 19651138 0 28 theme domain 46:51 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the GluR2 amino-terminal domain 0:51 Crystal structure of the GluR2 amino-terminal domain provides insights into the architecture and assembly of ionotropic glutamate receptors. 19651138 2 29 theme GluR2-ATD 340:348 arg1 structure 321:329 a 1.8-A resolution crystal structure 294:329 a 1.8-A resolution crystal structure of human GluR2-ATD 294:348 We report a 1.8-A resolution crystal structure of human GluR2-ATD. 19651138 4 30 theme full-length 647:657 arg1 receptors 664:672 full-length AMPA receptors 647:672 full-length AMPA receptors 647:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 19651138 2 31 theme human 334:338 arg1 GluR2-ATD 340:348 human GluR2-ATD 334:348 human GluR2-ATD 334:348 We report a 1.8-A resolution crystal structure of human GluR2-ATD. 19651138 1 32 theme 400-residue 243:253 arg1 ATD 278:280 ATD 278:280 ATD 278:280 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 1 32 theme 400-residue 243:253 arg1 domain 270:275 the 400-residue amino-terminal domain 239:275 the 400-residue amino-terminal domain (ATD) 239:281 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 1 33 theme glutamate 152:160 arg1 receptors 162:170 Ionotropic glutamate receptors 141:170 Ionotropic glutamate receptors 141:170 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 1 34 theme amino-terminal 255:268 arg1 ATD 278:280 ATD 278:280 ATD 278:280 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 1 34 theme amino-terminal 255:268 arg1 domain 270:275 the 400-residue amino-terminal domain 239:275 the 400-residue amino-terminal domain (ATD) 239:281 Ionotropic glutamate receptors are functionally diverse but have a common architecture, including the 400-residue amino-terminal domain (ATD). 19651138 4 35 theme AMPA 659:662 arg1 receptors 664:672 full-length AMPA receptors 647:672 full-length AMPA receptors 647:672 Lattice contacts in a 4.1-A resolution crystal form reveal a tetrameric (dimer-dimer) arrangement consistent with previous cellular and cryo-electron microscopic data for full-length AMPA receptors. 12855696 0 0 theme thioesterases 142:154 arg1 specificities 107:119 divergent substrate specificities 87:119 divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2 87:169 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 6 1 theme PPT1 1159:1162 arg1 structures 1145:1154 the structures 1141:1154 the structures of PPT1 and -2 1141:1169 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 2 attach present 1062:1068 arg2 we 1059:1060 we 1059:1060 we 1059:1060 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 2 attach present 1062:1068 arg1 paper 1052:1056 the current paper 1040:1056 the current paper 1040:1056 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 1 3 theme granular 443:450 arg1 deposits 452:459 amorphous granular deposits 433:459 amorphous granular deposits 433:459 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 7 4 theme located 1398:1404 arg1 beta8-alphaF 1384:1395 beta8-alphaF 1384:1395 beta8-alphaF 1384:1395 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 4 theme located 1398:1404 arg1 beta3-alphaA 1367:1378 beta3-alphaA 1367:1378 beta3-alphaA 1367:1378 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 4 theme located 1398:1404 arg1 loops 1360:1364 two parallel loops 1347:1364 two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove 1347:1447 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 6 5 dep A. 1123:1124 arg1 show 1171:1174 show 1171:1174 show very similar architectural features 1171:1210 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 3 6 theme preferred 691:699 arg1 substrate 701:709 the preferred substrate 687:709 the preferred substrate of PPT2 687:717 A previous study had suggested that palmitoyl-CoA was the preferred substrate of PPT2. 12855696 3 6 theme preferred 691:699 arg1 palmitoyl-CoA 669:681 palmitoyl-CoA 669:681 palmitoyl-CoA 669:681 A previous study had suggested that palmitoyl-CoA was the preferred substrate of PPT2. 12855696 6 7 theme conformational 1222:1235 arg1 differences 1237:1247 conformational differences 1222:1247 conformational differences in helix alpha4 1222:1263 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 5 8 theme recent 855:860 arg1 model 879:883 a recent transgenic mouse model 853:883 a recent transgenic mouse model 853:883 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 2 9 theme 26 609:610 arg1 % 611:611 % 611:611 % 611:611 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 0 10 theme lysosomal 132:140 arg1 thioesterases 142:154 the two lysosomal thioesterases 124:154 the two lysosomal thioesterases 124:154 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 0 10 theme lysosomal 132:140 arg1 PPT2 166:169 PPT2 166:169 PPT2 166:169 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 0 10 theme lysosomal 132:140 arg1 PPT1 157:160 PPT1 157:160 PPT1 157:160 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 5 11 theme mouse 873:877 arg1 model 879:883 a recent transgenic mouse model 853:883 a recent transgenic mouse model 853:883 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 6 12 theme similar 1181:1187 arg1 features 1203:1210 very similar architectural features 1176:1210 very similar architectural features 1176:1210 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 13 theme A. 1123:1124 arg1 resolution 1105:1114 a resolution 1103:1114 a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1 1103:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 1 14 theme impaired 359:366 arg1 degradation 368:378 impaired degradation 359:378 impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 359:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 2 15 with identity 613:620 arg1 PPT1 627:630 PPT1 627:630 PPT1 627:630 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 5 16 theme PPT2 834:837 arg1 deficiency 839:848 PPT2 deficiency 834:848 PPT2 deficiency in a recent transgenic mouse model 834:883 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 5 17 from deficiency 839:848 arg1 model 879:883 a recent transgenic mouse model 853:883 a recent transgenic mouse model 853:883 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 2 18 theme second 568:573 arg1 hydrolase 585:593 a second lysosomal hydrolase 566:593 a second lysosomal hydrolase that shares a 26% identity with PPT1 566:630 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 2 18 theme second 568:573 arg1 thioesterase-2 541:554 Palmitoyl protein thioesterase-2 523:554 Palmitoyl protein thioesterase-2 (PPT2) 523:561 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 6 19 from groove 1305:1310 arg1 PPT1 1315:1318 PPT1 1315:1318 PPT1 1315:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 20 theme architectural 1189:1201 arg1 features 1203:1210 very similar architectural features 1176:1210 very similar architectural features 1176:1210 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 21 theme solvent-exposed 1275:1289 arg1 groove 1305:1310 a solvent-exposed lipid-binding groove 1273:1310 a solvent-exposed lipid-binding groove in PPT1 1273:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 1 22 theme acid-modified 389:401 arg1 proteins 403:410 fatty acid-modified proteins 383:410 fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 383:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 4 23 from substrates 807:816 arg1 palmitate 756:764 palmitate 756:764 palmitate from the several S-palmitoylated protein substrates 756:816 Furthermore, PPT2 did not hydrolyze palmitate from the several S-palmitoylated protein substrates. 12855696 5 24 theme neuronal 914:921 arg1 lipofuscinosis 930:943 neuronal ceroid lipofuscinosis 914:943 neuronal ceroid lipofuscinosis 914:943 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 6 25 theme -2 1168:1169 arg1 structures 1145:1154 the structures 1141:1154 the structures of PPT1 and -2 1141:1169 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 1 26 from accumulation 417:428 arg1 neurons 473:479 cortical neurons 464:479 cortical neurons 464:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 2 27 theme Palmitoyl 523:531 arg1 PPT2 557:560 PPT2 557:560 PPT2 557:560 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 2 27 theme Palmitoyl 523:531 arg1 thioesterase-2 541:554 Palmitoyl protein thioesterase-2 523:554 Palmitoyl protein thioesterase-2 (PPT2) 523:561 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 2 27 theme Palmitoyl 523:531 arg1 hydrolase 585:593 a second lysosomal hydrolase 566:593 a second lysosomal hydrolase that shares a 26% identity with PPT1 566:630 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 1 28 theme storage 325:331 arg1 form 264:267 the infantile form 250:267 the infantile form 250:267 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 28 theme storage 325:331 arg1 disorder 333:340 a lysosomal storage disorder 313:340 a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 313:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 7 29 theme binding 1527:1533 arg1 groove 1535:1540 this binding groove 1522:1540 this binding groove 1522:1540 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 9 30 theme acid 1781:1784 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 7 31 dep loops 1360:1364 arg1 beta8-alphaF 1384:1395 beta8-alphaF 1384:1395 beta8-alphaF 1384:1395 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 31 dep loops 1360:1364 arg1 beta3-alphaA 1367:1378 beta3-alphaA 1367:1378 beta3-alphaA 1367:1378 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 31 dep loops 1360:1364 arg1 loops 1360:1364 two parallel loops 1347:1364 two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove 1347:1447 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 0 32 theme divergent 87:95 arg1 specificities 107:119 divergent substrate specificities 87:119 divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2 87:169 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 7 33 from space 1333:1337 arg1 PPT2 1452:1455 PPT2 1452:1455 PPT2 1452:1455 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 9 34 theme length 1792:1797 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 5 35 theme lipofuscinosis 930:943 arg1 form 906:909 a form 904:909 a form of neuronal ceroid lipofuscinosis 904:943 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 6 36 theme structures 1145:1154 arg1 Comparisons 1126:1136 Comparisons 1126:1136 Comparisons of the structures of PPT1 and -2 1126:1169 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 37 theme current 1044:1050 arg1 paper 1052:1056 the current paper 1040:1056 the current paper 1040:1056 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 0 38 theme palmitoyl 25:33 arg1 PPT2 59:62 PPT2 59:62 PPT2 59:62 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 0 38 theme palmitoyl 25:33 arg1 thioesterase-2 43:56 palmitoyl protein thioesterase-2 25:56 palmitoyl protein thioesterase-2 (PPT2) 25:63 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 1 39 theme neuronal 272:279 arg1 lipofuscinosis 288:301 neuronal ceroid lipofuscinosis 272:301 neuronal ceroid lipofuscinosis 272:301 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 7 40 theme head 1505:1508 arg1 groups 1510:1515 bulky head groups 1499:1515 bulky head groups 1499:1515 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 5 41 theme thioester 1015:1023 arg1 catabolism 1025:1034 lysosomal thioester catabolism 1005:1034 lysosomal thioester catabolism 1005:1034 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 0 42 theme thioesterase-2 43:56 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) 0:63 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 1 43 theme lipofuscinosis 288:301 arg1 form 264:267 the infantile form 250:267 the infantile form 250:267 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 43 theme lipofuscinosis 288:301 arg1 disorder 333:340 a lysosomal storage disorder 313:340 a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 313:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 6 44 from resolution 1105:1114 arg1 structure 1082:1090 the crystal structure 1070:1090 the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1 1070:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 9 45 from differences 1760:1770 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 7 46 theme fatty 1482:1486 arg1 acids 1488:1492 fatty acids 1482:1492 fatty acids with bulky head groups 1482:1515 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 5 47 theme non-redundant 982:994 arg1 roles 996:1000 non-redundant roles 982:1000 non-redundant roles 982:1000 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 4 48 theme S-palmitoylated 783:797 arg1 substrates 807:816 the several S-palmitoylated protein substrates 771:816 the several S-palmitoylated protein substrates 771:816 Furthermore, PPT2 did not hydrolyze palmitate from the several S-palmitoylated protein substrates. 12855696 8 49 theme structural 1580:1589 arg1 difference 1591:1600 This structural difference 1575:1600 This structural difference 1575:1600 This structural difference accounts for the ability of PPT2 to hydrolyze an unbranched structure such as palmitoyl-CoA but not palmitoylcysteine or palmitoylated proteins. 12855696 6 50 theme crystal 1074:1080 arg1 structure 1082:1090 the crystal structure 1070:1090 the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1 1070:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 6 51 theme helix 1252:1256 arg1 alpha4 1258:1263 helix alpha4 1252:1263 helix alpha4 1252:1263 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 1 52 theme amorphous 433:441 arg1 deposits 452:459 amorphous granular deposits 433:459 amorphous granular deposits 433:459 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 6 53 from differences 1237:1247 arg1 alpha4 1258:1263 helix alpha4 1252:1263 helix alpha4 1252:1263 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 2 54 theme lysosomal 575:583 arg1 hydrolase 585:593 a second lysosomal hydrolase 566:593 a second lysosomal hydrolase that shares a 26% identity with PPT1 566:630 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 2 54 theme lysosomal 575:583 arg1 thioesterase-2 541:554 Palmitoyl protein thioesterase-2 523:554 Palmitoyl protein thioesterase-2 (PPT2) 523:561 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 3 55 theme PPT2 714:717 arg1 substrate 701:709 the preferred substrate 687:709 the preferred substrate of PPT2 687:717 A previous study had suggested that palmitoyl-CoA was the preferred substrate of PPT2. 12855696 3 55 theme PPT2 714:717 arg1 palmitoyl-CoA 669:681 palmitoyl-CoA 669:681 palmitoyl-CoA 669:681 A previous study had suggested that palmitoyl-CoA was the preferred substrate of PPT2. 12855696 1 56 theme deposits 452:459 arg1 accumulation 417:428 accumulation 417:428 accumulation of amorphous granular deposits in cortical neurons 417:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 7 57 with acids 1488:1492 arg1 groups 1510:1515 bulky head groups 1499:1515 bulky head groups 1499:1515 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 58 theme lipid-binding 1428:1440 arg1 groove 1442:1447 the lipid-binding groove 1424:1447 the lipid-binding groove 1424:1447 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 2 59 theme % 611:611 arg1 identity 613:620 a 26% identity 607:620 a 26% identity with PPT1 607:630 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 1 60 theme cortical 464:471 arg1 neurons 473:479 cortical neurons 464:479 cortical neurons 464:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 5 61 theme transgenic 862:871 arg1 model 879:883 a recent transgenic mouse model 853:883 a recent transgenic mouse model 853:883 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 9 62 theme biochemical 1896:1906 arg1 basis 1908:1912 a biochemical basis 1894:1912 a biochemical basis for their non-redundant roles 1894:1942 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 8 63 theme palmitoylated 1723:1735 arg1 proteins 1737:1744 palmitoylated proteins 1723:1744 palmitoylated proteins 1723:1744 This structural difference accounts for the ability of PPT2 to hydrolyze an unbranched structure such as palmitoyl-CoA but not palmitoylcysteine or palmitoylated proteins. 12855696 1 64 theme protein 195:201 arg1 PPT1 219:222 PPT1 219:222 PPT1 219:222 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 64 theme protein 195:201 arg1 thioesterase-1 203:216 palmitoyl protein thioesterase-1 185:216 palmitoyl protein thioesterase-1 (PPT1) 185:223 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 2 65 theme protein 533:539 arg1 PPT2 557:560 PPT2 557:560 PPT2 557:560 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 2 65 theme protein 533:539 arg1 thioesterase-2 541:554 Palmitoyl protein thioesterase-2 523:554 Palmitoyl protein thioesterase-2 (PPT2) 523:561 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 2 65 theme protein 533:539 arg1 hydrolase 585:593 a second lysosomal hydrolase 566:593 a second lysosomal hydrolase that shares a 26% identity with PPT1 566:630 Palmitoyl protein thioesterase-2 (PPT2) is a second lysosomal hydrolase that shares a 26% identity with PPT1. 12855696 9 66 theme non-redundant 1924:1936 arg1 roles 1938:1942 their non-redundant roles 1918:1942 their non-redundant roles 1918:1942 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 6 67 theme lipid-binding 1291:1303 arg1 groove 1305:1310 a solvent-exposed lipid-binding groove 1273:1310 a solvent-exposed lipid-binding groove in PPT1 1273:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 1 68 theme fatty 383:387 arg1 proteins 403:410 fatty acid-modified proteins 383:410 fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 383:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 7 69 theme parallel 1351:1358 arg1 beta8-alphaF 1384:1395 beta8-alphaF 1384:1395 beta8-alphaF 1384:1395 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 69 theme parallel 1351:1358 arg1 beta3-alphaA 1367:1378 beta3-alphaA 1367:1378 beta3-alphaA 1367:1378 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 7 69 theme parallel 1351:1358 arg1 loops 1360:1364 two parallel loops 1347:1364 two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove 1347:1447 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 1 70 theme proteins 403:410 arg1 degradation 368:378 impaired degradation 359:378 impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 359:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 71 from Mutations 172:180 arg1 PPT1 219:222 PPT1 219:222 PPT1 219:222 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 71 from Mutations 172:180 arg1 thioesterase-1 203:216 palmitoyl protein thioesterase-1 185:216 palmitoyl protein thioesterase-1 (PPT1) 185:223 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 7 72 theme limited 1325:1331 arg1 space 1333:1337 The limited space 1321:1337 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 1321:1455 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 1 73 theme lysosomal 315:323 arg1 form 264:267 the infantile form 250:267 the infantile form 250:267 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 73 theme lysosomal 315:323 arg1 disorder 333:340 a lysosomal storage disorder 313:340 a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 313:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 9 74 theme fatty 1775:1779 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 0 75 theme substrate 97:105 arg1 specificities 107:119 divergent substrate specificities 87:119 divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2 87:169 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 1 76 theme palmitoyl 185:193 arg1 PPT1 219:222 PPT1 219:222 PPT1 219:222 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 76 theme palmitoyl 185:193 arg1 thioesterase-1 203:216 palmitoyl protein thioesterase-1 185:216 palmitoyl protein thioesterase-1 (PPT1) 185:223 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 9 77 theme chain 1786:1790 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 0 78 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) 0:63 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 5 79 theme ceroid 923:928 arg1 lipofuscinosis 930:943 neuronal ceroid lipofuscinosis 914:943 neuronal ceroid lipofuscinosis 914:943 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 1 80 with proteins 403:410 arg1 accumulation 417:428 accumulation 417:428 accumulation of amorphous granular deposits in cortical neurons 417:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 81 theme infantile 254:262 arg1 form 264:267 the infantile form 250:267 the infantile form 250:267 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 81 theme infantile 254:262 arg1 disorder 333:340 a lysosomal storage disorder 313:340 a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons 313:479 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 9 82 theme PPT1 1814:1817 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 0 83 theme protein 35:41 arg1 PPT2 59:62 PPT2 59:62 PPT2 59:62 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 0 83 theme protein 35:41 arg1 thioesterase-2 43:56 palmitoyl protein thioesterase-2 25:56 palmitoyl protein thioesterase-2 (PPT2) 25:63 The crystal structure of palmitoyl protein thioesterase-2 (PPT2) reveals the basis for divergent substrate specificities of the two lysosomal thioesterases, PPT1 and PPT2. 12855696 5 84 theme lysosomal 1005:1013 arg1 catabolism 1025:1034 lysosomal thioester catabolism 1005:1034 lysosomal thioester catabolism 1005:1034 Interestingly, PPT2 deficiency in a recent transgenic mouse model is associated with a form of neuronal ceroid lipofuscinosis, suggesting that PPT1 and -2 perform non-redundant roles in lysosomal thioester catabolism. 12855696 9 85 theme -2 1823:1824 arg1 specificity 1799:1809 fatty acid chain length specificity 1775:1809 fatty acid chain length specificity of PPT1 and -2 1775:1824 Furthermore, differences in fatty acid chain length specificity of PPT1 and -2, also reported here, are explained by the structure and may provide a biochemical basis for their non-redundant roles. 12855696 4 86 theme protein 799:805 arg1 substrates 807:816 the several S-palmitoylated protein substrates 771:816 the several S-palmitoylated protein substrates 771:816 Furthermore, PPT2 did not hydrolyze palmitate from the several S-palmitoylated protein substrates. 12855696 1 87 theme ceroid 281:286 arg1 lipofuscinosis 288:301 neuronal ceroid lipofuscinosis 272:301 neuronal ceroid lipofuscinosis 272:301 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 1 88 theme mental 493:498 arg1 retardation 500:510 mental retardation 493:510 mental retardation 493:510 Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. 12855696 8 89 theme PPT2 1630:1633 arg1 ability 1619:1625 the ability 1615:1625 the ability of PPT2 to hydrolyze an unbranched structure such as palmitoyl-CoA but not palmitoylcysteine or palmitoylated proteins 1615:1744 This structural difference accounts for the ability of PPT2 to hydrolyze an unbranched structure such as palmitoyl-CoA but not palmitoylcysteine or palmitoylated proteins. 12855696 7 90 theme bulky 1499:1503 arg1 groups 1510:1515 bulky head groups 1499:1515 bulky head groups 1499:1515 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 3 91 theme previous 635:642 arg1 study 644:648 A previous study 633:648 A previous study 633:648 A previous study had suggested that palmitoyl-CoA was the preferred substrate of PPT2. 12855696 7 92 theme acids 1488:1492 arg1 binding 1471:1477 the binding 1467:1477 the binding of fatty acids with bulky head groups 1467:1515 The limited space between two parallel loops (beta3-alphaA and beta8-alphaF) located immediately above the lipid-binding groove in PPT2 restricts the binding of fatty acids with bulky head groups, and this binding groove is significantly larger in PPT1. 12855696 6 93 theme PPT2 1095:1098 arg1 structure 1082:1090 the crystal structure 1070:1090 the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1 1070:1318 In the current paper, we present the crystal structure of PPT2 at a resolution of 2.7 A. Comparisons of the structures of PPT1 and -2 show very similar architectural features; however, conformational differences in helix alpha4 lead to a solvent-exposed lipid-binding groove in PPT1. 12855696 4 94 theme several 775:781 arg1 substrates 807:816 the several S-palmitoylated protein substrates 771:816 the several S-palmitoylated protein substrates 771:816 Furthermore, PPT2 did not hydrolyze palmitate from the several S-palmitoylated protein substrates. 12855696 8 95 theme unbranched 1651:1660 arg1 structure 1662:1670 an unbranched structure 1648:1670 an unbranched structure such as palmitoyl-CoA but not palmitoylcysteine or palmitoylated proteins 1648:1744 This structural difference accounts for the ability of PPT2 to hydrolyze an unbranched structure such as palmitoyl-CoA but not palmitoylcysteine or palmitoylated proteins. 18227435 5 0 dep acid 752:755 arg1 1 728:728 1 728:728 1 728:728 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 1 1 theme critical 168:175 arg1 factor 161:166 an oocyte-secreted factor 142:166 an oocyte-secreted factor critical for the regulation of ovarian physiology 142:216 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 1 theme critical 168:175 arg1 protein-15 119:128 Bone morphogenetic protein-15 100:128 Bone morphogenetic protein-15 (BMP-15) 100:137 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 0 2 theme protein 91:97 arg1 modification 43:54 the post-translational modification 20:54 the post-translational modification of recombinant human BMP-15 mature protein 20:97 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 3 3 from function 477:484 arg1 reproduction 506:517 female reproduction 499:517 female reproduction 499:517 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 2 4 theme protein 352:358 arg1 forms 360:364 two mature protein forms 341:364 two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) 341:411 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 6 5 theme important 1125:1133 arg1 contribution 1135:1146 an important contribution 1122:1146 an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility 1122:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 5 6 dep residue 806:812 arg1 2 795:795 2 795:795 2 795:795 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 2 7 theme SDS-PAGE 322:329 arg1 analysis 331:338 SDS-PAGE analysis 322:338 SDS-PAGE analysis 322:338 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 5 8 theme P16 935:937 arg1 acid 927:930 the C-terminal amino acid 906:930 (4) the C-terminal amino acid of P16 and P17 902:945 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 6 9 theme structure 1007:1015 arg1 knowledge 990:998 the first knowledge 980:998 the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function 980:1102 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 6 9 theme structure 1007:1015 arg1 findings 967:974 These findings 961:974 These findings 961:974 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 3 10 dep relevance 454:462 arg1 the 436:438 the 436:438 the 436:438 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 5 11 from Thr10 891:895 arg1 O-glycosylated 873:886 O-glycosylated 873:886 O-glycosylated 873:886 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 12 theme sixth 821:825 arg1 position 827:834 the sixth position 817:834 the sixth position of P16 817:841 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 6 13 theme growth 1215:1220 arg1 factors 1222:1228 oocyte-specific growth factors 1199:1228 oocyte-specific growth factors in modulation of female fertility 1199:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 2 14 theme kidney 288:293 arg1 cells 299:303 human embryonic kidney 293 cells 272:303 human embryonic kidney 293 cells 272:303 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 5 15 theme pyroglutamic 775:786 arg1 acid 788:791 pyroglutamic acid 775:791 pyroglutamic acid 775:791 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 15 theme pyroglutamic 775:786 arg1 acid 752:755 the N-terminal amino acid 731:755 (1) the N-terminal amino acid of P16 and P17 727:770 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 6 16 theme oocyte-specific 1199:1213 arg1 factors 1222:1228 oocyte-specific growth factors 1199:1228 oocyte-specific growth factors in modulation of female fertility 1199:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 5 17 theme P17 943:945 arg1 acid 927:930 the C-terminal amino acid 906:930 (4) the C-terminal amino acid of P16 and P17 902:945 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 6 18 theme function 1095:1102 arg1 basis 1079:1083 the molecular basis 1065:1083 the molecular basis of BMP-15 function 1065:1102 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 2 19 theme embryonic 278:286 arg1 kidney 288:293 human embryonic kidney 293 272:297 human embryonic kidney 293 cells 272:303 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 5 20 theme N-terminal 735:744 arg1 acid 788:791 pyroglutamic acid 775:791 pyroglutamic acid 775:791 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 20 theme N-terminal 735:744 arg1 acid 752:755 the N-terminal amino acid 731:755 (1) the N-terminal amino acid of P16 and P17 727:770 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 6 21 from factors 1222:1228 arg1 modulation 1233:1242 modulation 1233:1242 modulation of female fertility 1233:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 6 22 theme BMP-15 1088:1093 arg1 function 1095:1102 BMP-15 function 1088:1102 BMP-15 function 1088:1102 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 3 23 from relevance 454:462 arg1 reproduction 506:517 female reproduction 499:517 female reproduction 499:517 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 1 24 theme Bone 100:103 arg1 factor 161:166 an oocyte-secreted factor 142:166 an oocyte-secreted factor critical for the regulation of ovarian physiology 142:216 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 24 theme Bone 100:103 arg1 BMP-15 131:136 BMP-15 131:136 BMP-15 131:136 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 24 theme Bone 100:103 arg1 protein-15 119:128 Bone morphogenetic protein-15 100:128 Bone morphogenetic protein-15 (BMP-15) 100:137 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 4 25 theme proteins 629:636 arg1 structure 596:604 the structure 592:604 the structure of the rhBMP-15 mature proteins (P16 and P17) 592:650 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 5 26 theme amino 746:750 arg1 acid 788:791 pyroglutamic acid 775:791 pyroglutamic acid 775:791 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 26 theme amino 746:750 arg1 acid 752:755 the N-terminal amino acid 731:755 (1) the N-terminal amino acid of P16 and P17 727:770 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 1 27 theme morphogenetic 105:117 arg1 factor 161:166 an oocyte-secreted factor 142:166 an oocyte-secreted factor critical for the regulation of ovarian physiology 142:216 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 27 theme morphogenetic 105:117 arg1 BMP-15 131:136 BMP-15 131:136 BMP-15 131:136 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 27 theme morphogenetic 105:117 arg1 protein-15 119:128 Bone morphogenetic protein-15 100:128 Bone morphogenetic protein-15 (BMP-15) 100:137 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 28 theme ovarian 199:205 arg1 physiology 207:216 ovarian physiology 199:216 ovarian physiology 199:216 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 0 29 theme post-translational 24:41 arg1 modification 43:54 the post-translational modification 20:54 the post-translational modification of recombinant human BMP-15 mature protein 20:97 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 5 30 theme P16 839:841 arg1 position 827:834 the sixth position 817:834 the sixth position of P16 817:841 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 1 31 theme physiology 207:216 arg1 regulation 185:194 the regulation 181:194 the regulation of ovarian physiology 181:216 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 6 32 theme first 984:988 arg1 knowledge 990:998 the first knowledge 980:998 the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function 980:1102 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 6 32 theme first 984:988 arg1 findings 967:974 These findings 961:974 These findings 961:974 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 3 33 theme female 499:504 arg1 reproduction 506:517 female reproduction 499:517 female reproduction 499:517 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 5 34 theme C-terminal 910:919 arg1 acid 927:930 the C-terminal amino acid 906:930 (4) the C-terminal amino acid of P16 and P17 902:945 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 35 theme Ser 802:804 arg1 residue 806:812 the Ser residue 798:812 (2) the Ser residue at the sixth position of P16 794:841 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 36 gly O-glycosylated 873:886 arg2 Thr10 891:895 Thr10 891:895 Thr10 891:895 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 36 gly O-glycosylated 873:886 arg1 P17 866:868 (3) P17 862:868 (3) P17 862:868 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 3 37 theme BMP-15 489:494 arg1 relevance 454:462 physiological relevance 440:462 physiological relevance 440:462 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 3 37 theme BMP-15 489:494 arg1 function 477:484 critical function 468:484 critical function 468:484 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 6 38 theme fertility 1254:1262 arg1 modulation 1233:1242 modulation 1233:1242 modulation of female fertility 1233:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 6 39 theme research 1175:1182 arg1 area 1184:1187 the rapidly progressing research area 1151:1187 the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility 1151:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 5 40 theme amino 921:925 arg1 acid 927:930 the C-terminal amino acid 906:930 (4) the C-terminal amino acid of P16 and P17 902:945 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 3 41 theme rhBMP-15 559:566 arg1 structure 546:554 the structure 542:554 the structure of rhBMP-15 542:566 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 4 42 theme mature 622:627 arg1 P17 647:649 P17 647:649 P17 647:649 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 4 42 theme mature 622:627 arg1 P16 639:641 P16 639:641 P16 639:641 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 4 42 theme mature 622:627 arg1 proteins 629:636 the rhBMP-15 mature proteins 609:636 the rhBMP-15 mature proteins (P16 and P17) 609:650 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 5 43 theme P17 768:770 arg1 acid 788:791 pyroglutamic acid 775:791 pyroglutamic acid 775:791 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 43 theme P17 768:770 arg1 acid 752:755 the N-terminal amino acid 731:755 (1) the N-terminal amino acid of P16 and P17 727:770 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 0 44 theme modification 43:54 arg1 Characterization 0:15 Characterization 0:15 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 0:98 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 6 45 theme progressing 1163:1173 arg1 area 1184:1187 the rapidly progressing research area 1151:1187 the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility 1151:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 6 46 theme molecular 1069:1077 arg1 basis 1079:1083 the molecular basis 1065:1083 the molecular basis of BMP-15 function 1065:1102 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 5 47 dep P17 866:868 arg1 3 863:863 3 863:863 3 863:863 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 0 48 theme human 71:75 arg1 protein 91:97 recombinant human BMP-15 mature protein 59:97 recombinant human BMP-15 mature protein 59:97 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 6 49 theme female 1247:1252 arg1 fertility 1254:1262 female fertility 1247:1262 female fertility 1247:1262 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 2 50 theme mature 345:350 arg1 forms 360:364 two mature protein forms 341:364 two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) 341:411 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 0 51 theme recombinant 59:69 arg1 protein 91:97 recombinant human BMP-15 mature protein 59:97 recombinant human BMP-15 mature protein 59:97 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 2 52 theme recombinant 224:234 arg1 rhBMP-15 250:257 rhBMP-15 250:257 rhBMP-15 250:257 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 2 52 theme recombinant 224:234 arg1 BMP-15 242:247 recombinant human BMP-15 224:247 recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells 224:303 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 2 53 theme human 236:240 arg1 rhBMP-15 250:257 rhBMP-15 250:257 rhBMP-15 250:257 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 2 53 theme human 236:240 arg1 BMP-15 242:247 recombinant human BMP-15 224:247 recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells 224:303 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 3 54 theme critical 468:475 arg1 function 477:484 critical function 468:484 critical function 468:484 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 4 55 dep proteins 629:636 arg1 P17 647:649 P17 647:649 P17 647:649 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 4 55 dep proteins 629:636 arg1 P16 639:641 P16 639:641 P16 639:641 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 4 55 dep proteins 629:636 arg1 proteins 629:636 the rhBMP-15 mature proteins 609:636 the rhBMP-15 mature proteins (P16 and P17) 609:650 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 2 56 theme human 272:276 arg1 kidney 288:293 human embryonic kidney 293 272:297 human embryonic kidney 293 cells 272:303 When recombinant human BMP-15 (rhBMP-15) produced in human embryonic kidney 293 cells was subjected to SDS-PAGE analysis, two mature protein forms corresponding to 16 kDa (P16) and 17 kDa (P17) were observed. 18227435 3 57 theme physiological 440:452 arg1 relevance 454:462 physiological relevance 440:462 physiological relevance 440:462 Despite the physiological relevance and critical function of BMP-15 in female reproduction, little is known about the structure of rhBMP-15. 18227435 5 58 dep acid 927:930 arg1 4 903:903 4 903:903 4 903:903 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 6 59 theme protein 1036:1042 arg1 structure 1007:1015 the structure 1003:1015 the structure of rhBMP-15 mature protein 1003:1042 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 4 60 theme state-of-the-art 658:673 arg1 technology 686:695 state-of-the-art proteomics technology 658:695 state-of-the-art proteomics technology 658:695 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 5 61 from position 827:834 arg1 residue 806:812 the Ser residue 798:812 (2) the Ser residue at the sixth position of P16 794:841 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 4 62 theme proteomics 675:684 arg1 technology 686:695 state-of-the-art proteomics technology 658:695 state-of-the-art proteomics technology 658:695 Here, we have analyzed the structure of the rhBMP-15 mature proteins (P16 and P17) using state-of-the-art proteomics technology. 18227435 0 63 theme mature 84:89 arg1 protein 91:97 recombinant human BMP-15 mature protein 59:97 recombinant human BMP-15 mature protein 59:97 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 6 64 theme mature 1029:1034 arg1 protein 1036:1042 rhBMP-15 mature protein 1020:1042 rhBMP-15 mature protein 1020:1042 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 18227435 5 65 theme P16 760:762 arg1 acid 788:791 pyroglutamic acid 775:791 pyroglutamic acid 775:791 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 5 65 theme P16 760:762 arg1 acid 752:755 the N-terminal amino acid 731:755 (1) the N-terminal amino acid of P16 and P17 727:770 Our findings are as follows: (1) the N-terminal amino acid of P16 and P17 is pyroglutamic acid; (2) the Ser residue at the sixth position of P16 is phosphorylated; (3) P17 is O-glycosylated at Thr10; and (4) the C-terminal amino acid of P16 and P17 is truncated. 18227435 1 66 theme oocyte-secreted 145:159 arg1 factor 161:166 an oocyte-secreted factor 142:166 an oocyte-secreted factor critical for the regulation of ovarian physiology 142:216 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 1 66 theme oocyte-secreted 145:159 arg1 protein-15 119:128 Bone morphogenetic protein-15 100:128 Bone morphogenetic protein-15 (BMP-15) 100:137 Bone morphogenetic protein-15 (BMP-15) is an oocyte-secreted factor critical for the regulation of ovarian physiology. 18227435 0 67 theme BMP-15 77:82 arg1 protein 91:97 recombinant human BMP-15 mature protein 59:97 recombinant human BMP-15 mature protein 59:97 Characterization of the post-translational modification of recombinant human BMP-15 mature protein. 18227435 6 68 theme rhBMP-15 1020:1027 arg1 protein 1036:1042 rhBMP-15 mature protein 1020:1042 rhBMP-15 mature protein 1020:1042 These findings are the first knowledge of the structure of rhBMP-15 mature protein toward understanding the molecular basis of BMP-15 function and could provide an important contribution to the rapidly progressing research area involving oocyte-specific growth factors in modulation of female fertility. 16679516 12 0 theme thyroid 1991:1997 arg1 disease 1999:2005 autoimmune thyroid disease 1980:2005 autoimmune thyroid disease 1980:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 4 1 theme hTg-CS 598:603 arg1 fraction 586:593 the fraction 582:593 the fraction of hTg-CS in total hTg 582:616 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 12 2 theme thyroidal 1932:1940 arg1 manifestations 1962:1975 thyroidal and extra-thyroidal manifestations 1932:1975 thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease 1932:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 0 3 from terminus 167:174 arg1 accessibility 137:149 proteolytic accessibility 125:149 proteolytic accessibility at the carboxyl terminus 125:174 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 5 4 theme electrophoretic 663:677 arg1 shift 688:692 the electrophoretic mobility shift 659:692 the electrophoretic mobility shift 659:692 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 2 5 theme oligosaccharide 295:309 arg1 addition 316:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 5 6 theme hTg 791:793 arg1 proteolysis 776:786 limited proteolysis 768:786 limited proteolysis of hTg 768:793 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 6 7 theme chondroitin 905:915 arg1 nonapeptide 938:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 12 8 dep ability 1858:1864 arg1 function 1876:1883 function 1876:1883 to function as an autoantigen 1873:1901 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 5 9 theme residue 882:888 arg1 2514 890:893 residue 2514 882:893 residue 2514 882:893 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 11 10 theme regulation 1751:1760 arg1 mechanism 1738:1746 the molecular mechanism 1724:1746 the molecular mechanism of regulation of the hormonogenic efficiency 1724:1791 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 2 11 theme human 328:332 arg1 hTg 349:351 hTg 349:351 hTg 349:351 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 2 11 theme human 328:332 arg1 thyroglobulin 334:346 human thyroglobulin 328:346 human thyroglobulin (hTg) 328:352 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 6 12 theme digestion 997:1005 arg1 products 985:992 the products 981:992 the products of digestion of hTg with endoproteinase Glu-C 981:1038 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 11 13 from insights 1710:1717 arg1 hTg 1819:1821 hTg 1819:1821 hTg 1819:1821 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 0 14 theme proteolytic 125:135 arg1 accessibility 137:149 proteolytic accessibility 125:149 proteolytic accessibility at the carboxyl terminus 125:174 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 2 15 theme addition 316:323 arg1 site 256:259 the site 252:259 the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition 252:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 2 15 theme addition 316:323 arg1 addition 316:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 11 16 theme efficiency 1782:1791 arg1 regulation 1751:1760 regulation 1751:1760 regulation of the hormonogenic efficiency 1751:1791 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 7 17 theme in 1167:1168 arg1 assay 1176:1180 an in vitro assay 1164:1180 an in vitro assay of enzymatic iodination 1164:1204 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 10 18 theme 6-sulfate 1536:1544 arg1 unit 1562:1565 the chondroitin 6-sulfate oligosaccharide unit 1520:1565 the chondroitin 6-sulfate oligosaccharide unit of hTg-CS 1520:1575 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 0 19 theme carboxyl 158:165 arg1 terminus 167:174 the carboxyl terminus 154:174 the carboxyl terminus 154:174 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 9 20 theme molecules 1430:1438 arg1 subset 1416:1421 a subset 1414:1421 a subset of hTg molecules 1414:1438 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 6 21 theme pure 966:969 arg1 form 971:974 pure form 966:974 pure form 966:974 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 5 22 theme proteolysis 776:786 arg1 products 756:763 the products 752:763 the products of limited proteolysis of hTg 752:793 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 2 23 theme type 264:267 arg1 D 269:269 type D 264:269 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 2 23 theme type 264:267 arg1 6-sulfate 284:292 chondroitin 6-sulfate 272:292 chondroitin 6-sulfate 272:292 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 7 24 dep in 1167:1168 arg1 vitro 1170:1174 vitro 1170:1174 vitro 1170:1174 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 0 25 theme thyroglobulin 73:85 arg1 Ser-2730 55:62 Ser-2730 55:62 Ser-2730 of human thyroglobulin 55:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 1 26 theme Potential 177:185 arg1 insights 187:194 Potential insights 177:194 Potential insights into thyroid homeostasis and autoimmunity 177:236 Potential insights into thyroid homeostasis and autoimmunity. 16679516 6 27 theme endoproteinase 1019:1032 arg1 Glu-C 1034:1038 endoproteinase Glu-C 1019:1038 endoproteinase Glu-C 1019:1038 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 11 28 from hTg 1819:1821 arg1 insights 1710:1717 insights 1710:1717 insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg 1710:1821 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 0 29 theme hormone 96:102 arg1 formation 104:112 hormone formation 96:112 hormone formation 96:112 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 12 30 theme hTg 1869:1871 arg1 ability 1858:1864 the ability 1854:1864 the ability of hTg to function as an autoantigen 1854:1901 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 6 31 theme hTg 1010:1012 arg1 digestion 997:1005 digestion 997:1005 digestion of hTg with endoproteinase Glu-C 997:1038 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 6 32 dep LTAGXGLRE 1076:1084 arg1 residues 1087:1094 residues 2726-2734 1087:1104 residues 2726-2734 1087:1104 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 6 32 dep LTAGXGLRE 1076:1084 arg1 X 1107:1107 X 1107:1107 X being Ser2730 linked to the oligosaccharide chain 1107:1157 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 3 33 theme hTg-CS0 474:480 arg1 molecules 483:491 chondroitin 6-sulfate-devoid (hTg-CS0) molecules 444:491 chondroitin 6-sulfate-devoid (hTg-CS0) molecules 444:491 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 7 34 theme iodination 1195:1204 arg1 assay 1176:1180 an in vitro assay 1164:1180 an in vitro assay of enzymatic iodination 1164:1204 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 12 35 theme potential 1828:1836 arg1 implications 1838:1849 The potential implications 1824:1849 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease 1824:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 12 36 theme extra-thyroidal 1946:1960 arg1 manifestations 1962:1975 thyroidal and extra-thyroidal manifestations 1932:1975 thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease 1932:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 0 37 theme human 67:71 arg1 thyroglobulin 73:85 human thyroglobulin 67:85 human thyroglobulin 67:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 3 38 theme 6-sulfate-devoid 456:471 arg1 molecules 483:491 chondroitin 6-sulfate-devoid (hTg-CS0) molecules 444:491 chondroitin 6-sulfate-devoid (hTg-CS0) molecules 444:491 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 9 39 theme T3 1493:1494 arg1 formation 1496:1504 T3 formation 1493:1504 T3 formation 1493:1504 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 4 40 theme preparations 568:579 arg1 number 554:559 an ample number 545:559 an ample number of hTg preparations 545:579 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 11 41 from ratio 1810:1814 arg1 hTg 1819:1821 hTg 1819:1821 hTg 1819:1821 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 6 42 attach isolated 954:961 arg2 nonapeptide 938:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 6 42 attach isolated 954:961 arg1 products 985:992 the products 981:992 the products of digestion of hTg with endoproteinase Glu-C 981:1038 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 0 43 theme chondroitin 9:19 arg1 unit 47:50 A single chondroitin 6-sulfate oligosaccharide unit 0:50 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin 0:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 7 44 theme 3,5,3 1281:1285 arg1 yields 1230:1235 higher yields 1223:1235 higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) 1223:1320 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 6 45 attach linked 1123:1128 arg1 chain 1153:1157 the oligosaccharide chain 1133:1157 the oligosaccharide chain 1133:1157 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 6 45 attach linked 1123:1128 arg2 Ser2730 1115:1121 Ser2730 1115:1121 Ser2730 linked to the oligosaccharide chain 1115:1157 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 8 46 theme Unfractionated 1336:1349 arg1 hTg 1351:1353 Unfractionated hTg 1336:1353 Unfractionated hTg 1336:1353 Unfractionated hTg behaved as hTg-CS. 16679516 5 47 theme chondroitin 796:806 arg1 6-sulfate 808:816 chondroitin 6-sulfate 796:816 chondroitin 6-sulfate 796:816 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 0 48 theme oligosaccharide 31:45 arg1 unit 47:50 A single chondroitin 6-sulfate oligosaccharide unit 0:50 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin 0:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 10 49 with digestion 1653:1661 arg1 trypsin 1678:1684 trypsin 1678:1684 trypsin 1678:1684 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 4 50 theme ample 548:552 arg1 number 554:559 an ample number 545:559 an ample number of hTg preparations 545:579 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 3 51 theme D-glucuronic 515:526 arg1 content 533:539 their D-glucuronic acid content 509:539 their D-glucuronic acid content 509:539 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 6 52 theme oligosaccharide 1137:1151 arg1 chain 1153:1157 the oligosaccharide chain 1133:1157 the oligosaccharide chain 1133:1157 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 10 53 theme limited 1645:1651 arg1 digestion 1653:1661 the limited digestion 1641:1661 the limited digestion of hTg-CS with trypsin 1641:1684 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 9 54 theme chondroitin 1380:1390 arg1 6-sulfate 1392:1400 chondroitin 6-sulfate 1380:1400 chondroitin 6-sulfate addition to a subset of hTg molecules 1380:1438 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 12 55 theme manifestations 1962:1975 arg1 pathogenesis 1916:1927 the pathogenesis 1912:1927 the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease 1912:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 7 56 theme enzymatic 1185:1193 arg1 iodination 1195:1204 enzymatic iodination 1185:1204 enzymatic iodination 1185:1204 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 3 57 theme content 533:539 arg1 basis 500:504 the basis 496:504 the basis of their D-glucuronic acid content 496:539 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 12 58 theme autoimmune 1980:1989 arg1 disease 1999:2005 autoimmune thyroid disease 1980:2005 autoimmune thyroid disease 1980:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 2 59 theme unit 311:314 arg1 addition 316:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 6 60 theme 6-sulfate-containing 917:936 arg1 nonapeptide 938:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 11 61 theme T4/T3 1804:1808 arg1 ratio 1810:1814 the T4/T3 ratio 1800:1814 the T4/T3 ratio in hTg 1800:1821 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 9 62 theme 6-sulfate 1392:1400 arg1 addition 1402:1409 chondroitin 6-sulfate addition 1380:1409 chondroitin 6-sulfate addition to a subset of hTg molecules 1380:1438 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 6 63 theme single 898:903 arg1 nonapeptide 938:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide 896:948 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 6 64 with digestion 997:1005 arg1 Glu-C 1034:1038 endoproteinase Glu-C 1019:1038 endoproteinase Glu-C 1019:1038 A single chondroitin 6-sulfate-containing nonapeptide was isolated in pure form from the products of digestion of hTg with endoproteinase Glu-C, and its sequence was determined as LTAGXGLRE (residues 2726-2734, X being Ser2730 linked to the oligosaccharide chain). 16679516 10 65 theme chondroitin 1524:1534 arg1 unit 1562:1565 the chondroitin 6-sulfate oligosaccharide unit 1520:1565 the chondroitin 6-sulfate oligosaccharide unit of hTg-CS 1520:1575 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 4 66 from hTg 614:616 arg1 fraction 586:593 the fraction 582:593 the fraction of hTg-CS in total hTg 582:616 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 7 67 theme higher 1223:1228 arg1 yields 1230:1235 higher yields 1223:1235 higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) 1223:1320 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 1 68 theme thyroid 201:207 arg1 homeostasis 209:219 thyroid homeostasis 201:219 thyroid homeostasis 201:219 Potential insights into thyroid homeostasis and autoimmunity. 16679516 5 69 theme carboxyl-terminal 844:860 arg1 region 862:867 a carboxyl-terminal region 842:867 a carboxyl-terminal region 842:867 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 9 70 theme T4 1470:1471 arg1 level 1461:1465 the overall level 1449:1465 the overall level of T4 1449:1471 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 11 71 theme ratio 1810:1814 arg1 insights 1710:1717 insights 1710:1717 insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg 1710:1821 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 5 72 theme chondroitin 725:735 arg1 6-sulfate 737:745 chondroitin 6-sulfate 725:745 chondroitin 6-sulfate upon the products of limited proteolysis of hTg 725:793 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 10 73 theme oligosaccharide 1546:1560 arg1 unit 1562:1565 the chondroitin 6-sulfate oligosaccharide unit 1520:1565 the chondroitin 6-sulfate oligosaccharide unit of hTg-CS 1520:1575 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 5 74 theme mobility 679:686 arg1 shift 688:692 the electrophoretic mobility shift 659:692 the electrophoretic mobility shift 659:692 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 9 75 theme hTg 1426:1428 arg1 molecules 1430:1438 hTg molecules 1426:1438 hTg molecules 1426:1438 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 4 76 theme total 608:612 arg1 hTg 614:616 total hTg 608:616 total hTg 608:616 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 11 77 theme hormonogenic 1769:1780 arg1 efficiency 1782:1791 the hormonogenic efficiency 1765:1791 the hormonogenic efficiency 1765:1791 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 2 78 theme chondroitin 272:282 arg1 D 269:269 type D 264:269 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 2 78 theme chondroitin 272:282 arg1 6-sulfate 284:292 chondroitin 6-sulfate 272:292 chondroitin 6-sulfate 272:292 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 5 79 theme limited 768:774 arg1 proteolysis 776:786 limited proteolysis 768:786 limited proteolysis of hTg 768:793 By exploiting the electrophoretic mobility shift and metachromasia conferred by chondroitin 6-sulfate upon the products of limited proteolysis of hTg, chondroitin 6-sulfate was first restricted to a carboxyl-terminal region, starting at residue 2514. 16679516 2 80 theme D 269:269 arg1 addition 316:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 type D (chondroitin 6-sulfate) oligosaccharide unit addition 264:323 We localized the site of type D (chondroitin 6-sulfate) oligosaccharide unit addition to human thyroglobulin (hTg). 16679516 9 81 theme overall 1453:1459 arg1 level 1461:1465 the overall level 1449:1465 the overall level of T4 1449:1471 Thus, chondroitin 6-sulfate addition to a subset of hTg molecules enhanced the overall level of T4 and, in particular, T3 formation. 16679516 12 82 theme disease 1999:2005 arg1 manifestations 1962:1975 thyroidal and extra-thyroidal manifestations 1932:1975 thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease 1932:2005 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 10 83 theme bond 1595:1598 arg1 Lys2714-Gly2715 1600:1614 peptide bond Lys2714-Gly2715 1587:1614 peptide bond Lys2714-Gly2715 1587:1614 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 11 84 theme molecular 1728:1736 arg1 mechanism 1738:1746 the molecular mechanism 1724:1746 the molecular mechanism of regulation of the hormonogenic efficiency 1724:1791 These findings provide insights into the molecular mechanism of regulation of the hormonogenic efficiency and of the T4/T3 ratio in hTg. 16679516 12 85 from implications 1838:1849 arg1 ability 1858:1864 the ability 1854:1864 the ability of hTg to function as an autoantigen 1854:1901 The potential implications in the ability of hTg to function as an autoantigen and into the pathogenesis of thyroidal and extra-thyroidal manifestations of autoimmune thyroid disease are discussed. 16679516 0 86 from Ser-2730 55:62 arg1 unit 47:50 A single chondroitin 6-sulfate oligosaccharide unit 0:50 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin 0:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 4 87 from hTg-CS 598:603 arg1 hTg 614:616 total hTg 608:616 total hTg 608:616 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 10 88 theme peptide 1587:1593 arg1 Lys2714-Gly2715 1600:1614 peptide bond Lys2714-Gly2715 1587:1614 peptide bond Lys2714-Gly2715 1587:1614 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 0 89 theme single 2:7 arg1 unit 47:50 A single chondroitin 6-sulfate oligosaccharide unit 0:50 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin 0:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 10 90 theme hTg-CS 1570:1575 arg1 unit 1562:1565 the chondroitin 6-sulfate oligosaccharide unit 1520:1565 the chondroitin 6-sulfate oligosaccharide unit of hTg-CS 1520:1575 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 0 91 theme 6-sulfate 21:29 arg1 unit 47:50 A single chondroitin 6-sulfate oligosaccharide unit 0:50 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin 0:85 A single chondroitin 6-sulfate oligosaccharide unit at Ser-2730 of human thyroglobulin enhances hormone formation and limits proteolytic accessibility at the carboxyl terminus. 16679516 3 92 theme chondroitin 398:408 arg1 6-sulfate-containing 410:429 chondroitin 6-sulfate-containing 398:429 chondroitin 6-sulfate-containing (hTg-CS) 398:438 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 3 92 theme chondroitin 398:408 arg1 hTg-CS 432:437 hTg-CS 432:437 hTg-CS 432:437 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 7 93 theme -triiodothyronine 1247:1263 arg1 yields 1230:1235 higher yields 1223:1235 higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) 1223:1320 In an in vitro assay of enzymatic iodination, hTg-CS produced higher yields of 3,5,5 '-triiodothyronine (T3) (171%) and 3,5,3',5'-tetraiodothyronine (T4) (134%) than hTg-CS0. 16679516 3 94 theme chondroitin 444:454 arg1 molecules 483:491 chondroitin 6-sulfate-devoid (hTg-CS0) molecules 444:491 chondroitin 6-sulfate-devoid (hTg-CS0) molecules 444:491 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 4 95 dep 71.6 638:641 arg1 to 635:636 to 635:636 to 635:636 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 4 96 theme hTg 564:566 arg1 preparations 568:579 hTg preparations 564:579 hTg preparations 564:579 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 16679516 3 97 theme acid 528:531 arg1 content 533:539 their D-glucuronic acid content 509:539 their D-glucuronic acid content 509:539 hTg was chromatographically separated into chondroitin 6-sulfate-containing (hTg-CS) and chondroitin 6-sulfate-devoid (hTg-CS0) molecules on the basis of their D-glucuronic acid content. 16679516 10 98 theme hTg-CS 1666:1671 arg1 digestion 1653:1661 the limited digestion 1641:1661 the limited digestion of hTg-CS with trypsin 1641:1684 Furthermore, the chondroitin 6-sulfate oligosaccharide unit of hTg-CS protected peptide bond Lys2714-Gly2715 from proteolysis, during the limited digestion of hTg-CS with trypsin. 16679516 4 99 from fraction 586:593 arg1 hTg 614:616 total hTg 608:616 total hTg 608:616 In an ample number of hTg preparations, the fraction of hTg-CS in total hTg ranged from 32.0 to 71.6%. 25561468 0 0 theme β4-sheet-mediated 84:100 arg1 dimerization 102:113 β4-sheet-mediated dimerization 84:113 β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum 84:160 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 6 1 theme S508A/G553D 894:904 arg1 mutants 929:935 S508A/G553D and N506A/G553D double mutants 894:935 S508A/G553D and N506A/G553D double mutants 894:935 S508A/G553D and N506A/G553D double mutants dimerize but remain in the endoplasmic reticulum. 25561468 1 2 theme type 167:170 arg1 diabetes 174:181 type 1 diabetes 167:181 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 7 3 theme ICA512-ΔNTF 1065:1075 arg1 mutant 1083:1088 an ICA512-ΔNTF G553D mutant 1062:1088 an ICA512-ΔNTF G553D mutant 1062:1088 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 1 4 theme secretory 271:279 arg1 granules 281:288 the insulin secretory granules 259:288 the insulin secretory granules 259:288 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 10 5 theme constraint 1473:1482 arg1 Removal 1457:1463 Removal 1457:1463 Removal of this constraint 1457:1482 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 1 6 theme granules 281:288 arg1 ICA512/IA-2/RPTPN 195:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 6 theme granules 281:288 arg1 phosphatase 244:254 a receptor protein tyrosine phosphatase 216:254 a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail 216:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 6 theme granules 281:288 arg1 SGs 291:293 SGs 291:293 SGs 291:293 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 5 7 theme ME 665:666 arg1 N-glycosylation 685:699 ME ICA512 β2-strand N-glycosylation 665:699 ME ICA512 β2-strand N-glycosylation 665:699 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 7 8 theme fragment 1013:1020 arg1 Removal 987:993 Removal 987:993 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 987:1053 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 6 9 theme double 922:927 arg1 mutants 929:935 S508A/G553D and N506A/G553D double mutants 894:935 S508A/G553D and N506A/G553D double mutants 894:935 S508A/G553D and N506A/G553D double mutants dimerize but remain in the endoplasmic reticulum. 25561468 5 10 theme ICA512 668:673 arg1 N-glycosylation 685:699 ME ICA512 β2-strand N-glycosylation 665:699 ME ICA512 β2-strand N-glycosylation 665:699 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 4 11 theme ME 578:579 arg1 ICA512 581:586 ME ICA512 578:586 ME ICA512 578:586 Here we show that ME ICA512 prompts proICA512 dimerization in the endoplasmic reticulum. 25561468 0 12 theme ectodomain 122:131 arg1 dimerization 102:113 β4-sheet-mediated dimerization 84:113 β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum 84:160 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 6 13 theme N506A/G553D 910:920 arg1 mutants 929:935 S508A/G553D and N506A/G553D double mutants 894:935 S508A/G553D and N506A/G553D double mutants 894:935 S508A/G553D and N506A/G553D double mutants dimerize but remain in the endoplasmic reticulum. 25561468 2 14 theme region 425:430 arg1 role 399:402 The role 395:402 The role of its extracellular region 395:430 The role of its extracellular region remains unknown. 25561468 9 15 theme N506 1437:1440 arg1 glycosylation 1442:1454 N506 glycosylation 1437:1454 N506 glycosylation 1437:1454 Hence, we propose that the ME ICA512 β2-strand fosters proICA512 dimerization until NTF prevents N506 glycosylation. 25561468 1 16 theme diabetes 174:181 arg1 ICA512/IA-2/RPTPN 195:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 16 theme diabetes 174:181 arg1 phosphatase 244:254 a receptor protein tyrosine phosphatase 216:254 a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail 216:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 8 17 theme NTF 1238:1240 arg1 RESP18-HD 1266:1274 RESP18-HD 1266:1274 RESP18-HD 1266:1274 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 17 theme NTF 1238:1240 arg1 domain 1258:1263 the NTF RESP18 homology domain 1234:1263 the NTF RESP18 homology domain (RESP18-HD) 1234:1275 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 1 18 theme cytosolic 379:387 arg1 tail 389:392 its cytosolic tail 375:392 its cytosolic tail 375:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 3 19 from β4-strands 490:499 arg1 ectodomain 515:524 the mature ectodomain 504:524 the mature ectodomain (ME ICA512) 504:536 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 3 19 from β4-strands 490:499 arg1 ICA512 530:535 ME ICA512 527:535 ME ICA512 527:535 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 9 20 theme ME 1367:1368 arg1 β2-strand 1377:1385 the ME ICA512 β2-strand 1363:1385 the ME ICA512 β2-strand 1363:1385 Hence, we propose that the ME ICA512 β2-strand fosters proICA512 dimerization until NTF prevents N506 glycosylation. 25561468 1 21 theme autoantigen 183:193 arg1 ICA512/IA-2/RPTPN 195:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 21 theme autoantigen 183:193 arg1 phosphatase 244:254 a receptor protein tyrosine phosphatase 216:254 a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail 216:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 22 theme tail 389:392 arg1 cleavage/signaling 353:370 cleavage/signaling 353:370 cleavage/signaling of its cytosolic tail 353:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 7 23 theme endoplasmic 1102:1112 arg1 reticulum 1114:1122 the endoplasmic reticulum 1098:1122 the endoplasmic reticulum 1098:1122 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 7 24 theme G553D 1077:1081 arg1 mutant 1083:1088 an ICA512-ΔNTF G553D mutant 1062:1088 an ICA512-ΔNTF G553D mutant 1062:1088 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 8 25 theme SG 1205:1206 arg1 sorting 1208:1214 SG sorting 1205:1214 SG sorting 1205:1214 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 5 26 theme ME 873:874 arg1 β4-strand 883:891 the ME ICA512 β4-strand 869:891 the ME ICA512 β4-strand 869:891 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 10 27 from reticulum 1563:1571 arg1 dimerization 1523:1534 proICA512 β4-strand-induced dimerization 1495:1534 proICA512 β4-strand-induced dimerization 1495:1534 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 10 27 from reticulum 1563:1571 arg1 O-glycosylation 1574:1588 O-glycosylation 1574:1588 O-glycosylation 1574:1588 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 10 27 from reticulum 1563:1571 arg1 targeting 1614:1622 RESP18-HD-mediated targeting 1595:1622 RESP18-HD-mediated targeting to granules 1595:1634 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 10 27 from reticulum 1563:1571 arg1 exit 1537:1540 exit 1537:1540 exit from the endoplasmic reticulum 1537:1571 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 10 28 theme proICA512 1495:1503 arg1 dimerization 1523:1534 proICA512 β4-strand-induced dimerization 1495:1534 proICA512 β4-strand-induced dimerization 1495:1534 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 9 29 theme ICA512 1370:1375 arg1 β2-strand 1377:1385 the ME ICA512 β2-strand 1363:1385 the ME ICA512 β2-strand 1363:1385 Hence, we propose that the ME ICA512 β2-strand fosters proICA512 dimerization until NTF prevents N506 glycosylation. 25561468 4 30 theme endoplasmic 626:636 arg1 reticulum 638:646 the endoplasmic reticulum 622:646 the endoplasmic reticulum 622:646 Here we show that ME ICA512 prompts proICA512 dimerization in the endoplasmic reticulum. 25561468 0 31 theme proICA512/IA-2 13:26 arg1 targeting 36:44 its targeting 32:44 its targeting to insulin secretory granules 32:74 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 0 31 theme proICA512/IA-2 13:26 arg1 Stability 0:8 Stability 0:8 Stability of proICA512/IA-2 0:26 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 10 32 theme endoplasmic 1551:1561 arg1 reticulum 1563:1571 the endoplasmic reticulum 1547:1571 the endoplasmic reticulum 1547:1571 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 1 33 theme receptor 218:225 arg1 ICA512/IA-2/RPTPN 195:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 33 theme receptor 218:225 arg1 phosphatase 244:254 a receptor protein tyrosine phosphatase 216:254 a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail 216:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 33 theme receptor 218:225 arg1 SGs 291:293 SGs 291:293 SGs 291:293 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 7 34 theme cell 1176:1179 arg1 surface 1181:1187 the cell surface 1172:1187 the cell surface 1172:1187 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 10 35 theme β4-strand-induced 1505:1521 arg1 dimerization 1523:1534 proICA512 β4-strand-induced dimerization 1495:1534 proICA512 β4-strand-induced dimerization 1495:1534 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 2 36 theme extracellular 411:423 arg1 region 425:430 its extracellular region 407:430 its extracellular region 407:430 The role of its extracellular region remains unknown. 25561468 1 37 theme protein 227:233 arg1 ICA512/IA-2/RPTPN 195:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 37 theme protein 227:233 arg1 phosphatase 244:254 a receptor protein tyrosine phosphatase 216:254 a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail 216:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 37 theme protein 227:233 arg1 SGs 291:293 SGs 291:293 SGs 291:293 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 3 38 theme Structural 449:458 arg1 studies 460:466 Structural studies 449:466 Structural studies 449:466 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 0 39 theme endoplasmic 140:150 arg1 reticulum 152:160 the endoplasmic reticulum 136:160 the endoplasmic reticulum 136:160 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 8 40 theme soluble 1286:1292 arg1 NTF 1294:1296 soluble NTF 1286:1296 soluble NTF 1286:1296 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 7 41 theme ME 1045:1046 arg1 ICA512 1048:1053 ME ICA512 1045:1053 ME ICA512 1045:1053 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 1 42 theme tyrosine 235:242 arg1 ICA512/IA-2/RPTPN 195:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN 163:211 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 42 theme tyrosine 235:242 arg1 phosphatase 244:254 a receptor protein tyrosine phosphatase 216:254 a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail 216:392 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 1 42 theme tyrosine 235:242 arg1 SGs 291:293 SGs 291:293 SGs 291:293 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 0 43 theme insulin 49:55 arg1 granules 67:74 insulin secretory granules 49:74 insulin secretory granules 49:74 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 6 44 theme endoplasmic 964:974 arg1 reticulum 976:984 the endoplasmic reticulum 960:984 the endoplasmic reticulum 960:984 S508A/G553D and N506A/G553D double mutants dimerize but remain in the endoplasmic reticulum. 25561468 5 45 theme β2-strand 675:683 arg1 N-glycosylation 685:699 ME ICA512 β2-strand N-glycosylation 665:699 ME ICA512 β2-strand N-glycosylation 665:699 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 5 46 theme proICA512 735:743 arg1 dimerization 745:756 proICA512 dimerization 735:756 proICA512 dimerization 735:756 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 1 47 theme granule 324:330 arg1 stores 332:337 granule stores 324:337 granule stores 324:337 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 5 48 theme N-glycosylation 685:699 arg1 Perturbation 649:660 Perturbation 649:660 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement 649:722 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 8 49 located located 1219:1225 arg2 located 1219:1225 located 1219:1225 located 1219:1225 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 49 located located 1219:1225 arg2 signal 1194:1199 The signal 1190:1199 The signal for SG sorting 1190:1214 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 49 located located 1219:1225 arg1 RESP18-HD 1266:1274 RESP18-HD 1266:1274 RESP18-HD 1266:1274 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 49 located located 1219:1225 arg1 domain 1258:1263 the NTF RESP18 homology domain 1234:1263 the NTF RESP18 homology domain (RESP18-HD) 1234:1275 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 3 50 from β2- 483:485 arg1 ectodomain 515:524 the mature ectodomain 504:524 the mature ectodomain (ME ICA512) 504:536 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 3 50 from β2- 483:485 arg1 ICA512 530:535 ME ICA512 527:535 ME ICA512 527:535 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 3 51 theme mature 508:513 arg1 ectodomain 515:524 the mature ectodomain 504:524 the mature ectodomain (ME ICA512) 504:536 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 3 51 theme mature 508:513 arg1 ICA512 530:535 ME ICA512 527:535 ME ICA512 527:535 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 1 52 theme stores 332:337 arg1 size 316:319 the size 312:319 the size of granule stores 312:337 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 25561468 5 53 from replacement 854:864 arg1 β4-strand 883:891 the ME ICA512 β4-strand 869:891 the ME ICA512 β4-strand 869:891 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 4 54 theme proICA512 596:604 arg1 dimerization 606:617 proICA512 dimerization 596:617 proICA512 dimerization 596:617 Here we show that ME ICA512 prompts proICA512 dimerization in the endoplasmic reticulum. 25561468 5 55 theme S508A 706:710 arg1 replacement 712:722 S508A replacement 706:722 S508A replacement 706:722 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 8 56 theme endoplasmic 1317:1327 arg1 reticulum 1329:1337 the endoplasmic reticulum 1313:1337 the endoplasmic reticulum 1313:1337 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 57 theme RESP18 1242:1247 arg1 RESP18-HD 1266:1274 RESP18-HD 1266:1274 RESP18-HD 1266:1274 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 57 theme RESP18 1242:1247 arg1 domain 1258:1263 the NTF RESP18 homology domain 1234:1263 the NTF RESP18 homology domain (RESP18-HD) 1234:1275 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 0 58 from dimerization 102:113 arg1 reticulum 152:160 the endoplasmic reticulum 136:160 the endoplasmic reticulum 136:160 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 5 59 theme G553D 848:852 arg1 replacement 854:864 G553D replacement 848:864 G553D replacement in the ME ICA512 β4-strand 848:891 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 3 60 theme ME 527:528 arg1 ectodomain 515:524 the mature ectodomain 504:524 the mature ectodomain (ME ICA512) 504:536 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 3 60 theme ME 527:528 arg1 ICA512 530:535 ME ICA512 527:535 ME ICA512 527:535 Structural studies indicated that β2- or β4-strands in the mature ectodomain (ME ICA512) form dimers in vitro. 25561468 5 61 theme ICA512 876:881 arg1 β4-strand 883:891 the ME ICA512 β4-strand 869:891 the ME ICA512 β4-strand 869:891 Perturbation of ME ICA512 β2-strand N-glycosylation upon S508A replacement allows for proICA512 dimerization, O-glycosylation, targeting to granules, and conversion, which are instead precluded upon G553D replacement in the ME ICA512 β4-strand. 25561468 9 62 theme proICA512 1395:1403 arg1 dimerization 1405:1416 proICA512 dimerization 1395:1416 proICA512 dimerization 1395:1416 Hence, we propose that the ME ICA512 β2-strand fosters proICA512 dimerization until NTF prevents N506 glycosylation. 25561468 0 63 theme secretory 57:65 arg1 granules 67:74 insulin secretory granules 49:74 insulin secretory granules 49:74 Stability of proICA512/IA-2 and its targeting to insulin secretory granules require β4-sheet-mediated dimerization of its ectodomain in the endoplasmic reticulum. 25561468 8 64 theme homology 1249:1256 arg1 RESP18-HD 1266:1274 RESP18-HD 1266:1274 RESP18-HD 1266:1274 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 8 64 theme homology 1249:1256 arg1 domain 1258:1263 the NTF RESP18 homology domain 1234:1263 the NTF RESP18 homology domain (RESP18-HD) 1234:1275 The signal for SG sorting is located within the NTF RESP18 homology domain (RESP18-HD), whereas soluble NTF is retained in the endoplasmic reticulum. 25561468 10 65 theme RESP18-HD-mediated 1595:1612 arg1 targeting 1614:1622 RESP18-HD-mediated targeting 1595:1622 RESP18-HD-mediated targeting to granules 1595:1634 Removal of this constraint allows for proICA512 β4-strand-induced dimerization, exit from the endoplasmic reticulum, O-glycosylation, and RESP18-HD-mediated targeting to granules. 25561468 7 66 theme N-terminal 1002:1011 arg1 ICA512-NTF 1023:1032 ICA512-NTF 1023:1032 ICA512-NTF 1023:1032 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 7 66 theme N-terminal 1002:1011 arg1 fragment 1013:1020 the N-terminal fragment 998:1020 the N-terminal fragment (ICA512-NTF) preceding ME ICA512 998:1053 Removal of the N-terminal fragment (ICA512-NTF) preceding ME ICA512 allows an ICA512-ΔNTF G553D mutant to exit the endoplasmic reticulum, and ICA512-ΔNTF is constitutively delivered to the cell surface. 25561468 1 67 theme insulin 263:269 arg1 granules 281:288 the insulin secretory granules 259:288 the insulin secretory granules 259:288 The type 1 diabetes autoantigen ICA512/IA-2/RPTPN is a receptor protein tyrosine phosphatase of the insulin secretory granules (SGs) which regulates the size of granule stores, possibly via cleavage/signaling of its cytosolic tail. 9023546 12 0 with analogy 1850:1856 arg1 glycosyltransferases 1869:1888 other glycosyltransferases 1863:1888 other glycosyltransferases 1863:1888 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 9 1 theme product 1372:1378 arg1 characterization 1339:1354 characterization 1339:1354 characterization of the purified product by electrospray mass spectrometry 1339:1412 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 12 2 theme stem 1907:1910 arg1 region 1913:1918 a 'stem' region 1904:1918 a 'stem' region that is very susceptible to proteolysis 1904:1958 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 12 2 theme stem 1907:1910 arg1 susceptible 1933:1943 susceptible 1933:1943 susceptible 1933:1943 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 2 3 theme conserved 290:298 arg1 serine 300:305 a conserved serine or threonine residue 288:326 serine 300:305 The enzyme catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains. 9023546 9 4 theme electrospray 1383:1394 arg1 spectrometry 1401:1412 electrospray mass spectrometry 1383:1412 electrospray mass spectrometry 1383:1412 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 6 5 theme consensus 863:871 arg1 sequence 881:888 a consensus primary sequence 861:888 more than just a consensus primary sequence 846:888 Therefore, the enzyme appears to require more than just a consensus primary sequence and likely requires that the EGF domain disulfide bonds be properly formed. 9023546 4 6 theme acceptor 642:649 arg1 peptides 468:475 Synthetic peptides 458:475 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) 458:613 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 4 6 theme acceptor 642:649 arg1 substrates 651:660 efficient acceptor substrates 632:660 efficient acceptor substrates 632:660 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 12 7 dep protein 1834:1840 arg1 has 1900:1902 has 1900:1902 has a 'stem' region that is very susceptible to proteolysis 1900:1958 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 8 8 theme enzymatic 1143:1151 arg1 activity 1153:1160 enzymatic activity 1143:1160 enzymatic activity 1143:1160 Although the enzyme did not exhibit an absolute requirement for Mn2+, enzymatic activity did increase ten fold in the presence of 50 mM MnCl2. 9023546 8 9 theme 50 1203:1204 arg1 mM 1206:1207 mM 1206:1207 mM 1206:1207 Although the enzyme did not exhibit an absolute requirement for Mn2+, enzymatic activity did increase ten fold in the presence of 50 mM MnCl2. 9023546 10 10 theme CHO 1545:1547 arg1 homogenates 1554:1564 CHO cell homogenates 1545:1564 CHO cell homogenates 1545:1564 Most of the enzymatic activity was found to be in the soluble fraction of CHO cell homogenates. 9023546 8 11 theme MnCl2 1209:1213 arg1 presence 1191:1198 the presence 1187:1198 the presence of 50 mM MnCl2 1187:1213 Although the enzyme did not exhibit an absolute requirement for Mn2+, enzymatic activity did increase ten fold in the presence of 50 mM MnCl2. 9023546 6 12 theme disulfide 930:938 arg1 bonds 940:944 the EGF domain disulfide bonds 915:944 the EGF domain disulfide bonds 915:944 Therefore, the enzyme appears to require more than just a consensus primary sequence and likely requires that the EGF domain disulfide bonds be properly formed. 9023546 11 13 theme extensive 1749:1757 arg1 washes 1767:1772 extensive aqueous washes 1749:1772 extensive aqueous washes 1749:1772 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 5 14 theme EGF 716:718 arg1 domains 720:726 complete EGF domains 707:726 complete EGF domains 707:726 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 7 15 theme linear 996:1001 arg1 dependency 1003:1012 linear dependency 996:1012 linear dependency of its activity on time, amount of enzyme, and substrates 996:1070 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 6 16 theme EGF 919:921 arg1 bonds 940:944 the EGF domain disulfide bonds 915:944 the EGF domain disulfide bonds 915:944 Therefore, the enzyme appears to require more than just a consensus primary sequence and likely requires that the EGF domain disulfide bonds be properly formed. 9023546 10 17 theme homogenates 1554:1564 arg1 fraction 1533:1540 the soluble fraction 1521:1540 the soluble fraction of CHO cell homogenates 1521:1564 Most of the enzymatic activity was found to be in the soluble fraction of CHO cell homogenates. 9023546 11 18 theme particles 1733:1741 arg1 extraction 1706:1715 Triton X-100 extraction 1693:1715 Triton X-100 extraction of the membrane particles 1693:1741 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 5 19 theme synthetic 669:677 arg1 peptides 679:686 These synthetic peptides 663:686 These synthetic peptides 663:686 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 11 20 theme activity 1667:1674 arg1 activity 1667:1674 the activity 1663:1674 the activity 1663:1674 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 11 20 theme activity 1667:1674 arg1 % 1658:1658 37% 1656:1658 37% of the activity 1656:1674 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 2 21 theme O-glycosidic 264:275 arg1 linkage 277:283 an O-glycosidic linkage 261:283 an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains 261:341 The enzyme catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains. 9023546 2 22 from serine 300:305 arg1 domains 335:341 EGF domains 331:341 EGF domains 331:341 The enzyme catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains. 9023546 0 23 theme enzymatic 68:76 arg1 addition 78:85 enzymatic addition 68:85 enzymatic addition of O-linked fucose to EGF domains 68:119 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 9 24 theme in 1220:1221 arg1 reaction 1243:1250 The in vitro glycosylation reaction 1216:1250 The in vitro glycosylation reaction 1216:1250 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 11 25 theme Triton 1693:1698 arg1 extraction 1706:1715 Triton X-100 extraction 1693:1715 Triton X-100 extraction of the membrane particles 1693:1741 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 5 26 theme cysteine 756:763 arg1 residues 765:772 all six cysteine residues 748:772 all six cysteine residues that define the EGF structure 748:802 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 9 27 theme glycosylation 1229:1241 arg1 reaction 1243:1250 The in vitro glycosylation reaction 1216:1250 The in vitro glycosylation reaction 1216:1250 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 1 28 theme fucosyltransferase 157:174 arg1 assay 125:129 An assay 122:129 An assay of GDP-fucose:polypeptide fucosyltransferase 122:174 An assay of GDP-fucose:polypeptide fucosyltransferase has been established. 9023546 3 29 theme recombinant 359:369 arg1 VII 384:386 recombinant human factor VII 359:386 recombinant human factor VII EGF-1 domain 359:399 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 5 30 contain contain 740:746 arg1 peptides 679:686 These synthetic peptides 663:686 These synthetic peptides 663:686 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 5 30 contain contain 740:746 arg2 residues 765:772 all six cysteine residues 748:772 all six cysteine residues that define the EGF structure 748:802 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 0 31 theme fucose 99:104 arg1 fucosyltransferase 45:62 polypeptide fucosyltransferase 33:62 polypeptide fucosyltransferase 33:62 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 0 31 theme fucose 99:104 arg1 addition 78:85 enzymatic addition 68:85 enzymatic addition of O-linked fucose to EGF domains 68:119 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 3 32 theme donor 441:445 arg1 substrate 447:455 donor substrate 441:455 donor substrate 441:455 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 0 33 theme EGF 109:111 arg1 domains 113:119 EGF domains 109:119 EGF domains 109:119 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 9 34 theme complete 1264:1271 arg1 conversion 1273:1282 complete conversion 1264:1282 complete conversion of the acceptor substrate to glycosylated product 1264:1332 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 3 35 theme EGF-1 388:392 arg1 domain 394:399 recombinant human factor VII EGF-1 domain 359:399 recombinant human factor VII EGF-1 domain 359:399 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 9 36 gly glycosylated 1313:1324 arg1 product 1326:1332 glycosylated product 1313:1332 glycosylated product 1313:1332 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 3 37 theme factor 377:382 arg1 VII 384:386 recombinant human factor VII 359:386 recombinant human factor VII EGF-1 domain 359:399 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 9 38 theme acceptor 1291:1298 arg1 substrate 1300:1308 the acceptor substrate 1287:1308 the acceptor substrate 1287:1308 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 11 39 theme protease 1635:1642 arg1 inhibitors 1644:1653 protease inhibitors 1635:1653 protease inhibitors 1635:1653 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 0 40 link O-linked 90:97 arg1 fucose 99:104 O-linked fucose 90:104 O-linked fucose 90:104 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 10 41 theme enzymatic 1483:1491 arg1 activity 1493:1500 the enzymatic activity 1479:1500 the enzymatic activity 1479:1500 Most of the enzymatic activity was found to be in the soluble fraction of CHO cell homogenates. 9023546 6 42 dep more 846:849 arg1 sequence 881:888 a consensus primary sequence 861:888 more than just a consensus primary sequence 846:888 Therefore, the enzyme appears to require more than just a consensus primary sequence and likely requires that the EGF domain disulfide bonds be properly formed. 9023546 0 43 dep Identification 0:13 arg1 fucosyltransferase 45:62 polypeptide fucosyltransferase 33:62 polypeptide fucosyltransferase 33:62 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 0 43 dep Identification 0:13 arg1 addition 78:85 enzymatic addition 68:85 enzymatic addition of O-linked fucose to EGF domains 68:119 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 9 44 theme purified 1363:1370 arg1 product 1372:1378 the purified product 1359:1378 the purified product 1359:1378 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 10 45 theme soluble 1525:1531 arg1 fraction 1533:1540 the soluble fraction 1521:1540 the soluble fraction of CHO cell homogenates 1521:1564 Most of the enzymatic activity was found to be in the soluble fraction of CHO cell homogenates. 9023546 8 46 theme absolute 1112:1119 arg1 requirement 1121:1131 an absolute requirement 1109:1131 an absolute requirement for Mn2+ 1109:1140 Although the enzyme did not exhibit an absolute requirement for Mn2+, enzymatic activity did increase ten fold in the presence of 50 mM MnCl2. 9023546 6 47 theme primary 873:879 arg1 sequence 881:888 a consensus primary sequence 861:888 more than just a consensus primary sequence 846:888 Therefore, the enzyme appears to require more than just a consensus primary sequence and likely requires that the EGF domain disulfide bonds be properly formed. 9023546 9 48 theme mass 1396:1399 arg1 spectrometry 1401:1412 electrospray mass spectrometry 1383:1412 electrospray mass spectrometry 1383:1412 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 4 49 link O-linked 545:552 arg1 fucose 554:559 O-linked fucose 545:559 O-linked fucose 545:559 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 4 50 theme efficient 632:640 arg1 peptides 468:475 Synthetic peptides 458:475 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) 458:613 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 4 50 theme efficient 632:640 arg1 substrates 651:660 efficient acceptor substrates 632:660 efficient acceptor substrates 632:660 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 10 51 theme cell 1549:1552 arg1 homogenates 1554:1564 CHO cell homogenates 1545:1564 CHO cell homogenates 1545:1564 Most of the enzymatic activity was found to be in the soluble fraction of CHO cell homogenates. 9023546 11 52 theme rat 1606:1608 arg1 liver 1610:1614 rat liver 1606:1614 rat liver 1606:1614 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 7 53 theme enzyme 1049:1054 arg1 enzyme 1049:1054 enzyme 1049:1054 enzyme 1049:1054 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 7 53 theme enzyme 1049:1054 arg1 substrates 1061:1070 substrates 1061:1070 substrates 1061:1070 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 7 53 theme enzyme 1049:1054 arg1 amount 1039:1044 amount 1039:1044 amount of enzyme 1039:1054 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 7 53 theme enzyme 1049:1054 arg1 time 1033:1036 time 1033:1036 time 1033:1036 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 11 54 theme membrane 1724:1731 arg1 particles 1733:1741 the membrane particles 1720:1741 the membrane particles 1720:1741 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 2 55 theme threonine 310:318 arg1 residue 320:326 a conserved serine or threonine residue 288:326 residue 320:326 The enzyme catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains. 9023546 4 56 theme Synthetic 458:466 arg1 substrates 651:660 efficient acceptor substrates 632:660 efficient acceptor substrates 632:660 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 4 56 theme Synthetic 458:466 arg1 peptides 468:475 Synthetic peptides 458:475 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) 458:613 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 8 57 theme mM 1206:1207 arg1 MnCl2 1209:1213 50 mM MnCl2 1203:1213 50 mM MnCl2 1203:1213 Although the enzyme did not exhibit an absolute requirement for Mn2+, enzymatic activity did increase ten fold in the presence of 50 mM MnCl2. 9023546 5 58 theme complete 707:714 arg1 domains 720:726 complete EGF domains 707:726 complete EGF domains 707:726 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 6 59 theme domain 923:928 arg1 bonds 940:944 the EGF domain disulfide bonds 915:944 the EGF domain disulfide bonds 915:944 Therefore, the enzyme appears to require more than just a consensus primary sequence and likely requires that the EGF domain disulfide bonds be properly formed. 9023546 7 60 from dependency 1003:1012 arg1 enzyme 1049:1054 enzyme 1049:1054 enzyme 1049:1054 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 7 60 from dependency 1003:1012 arg1 substrates 1061:1070 substrates 1061:1070 substrates 1061:1070 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 7 60 from dependency 1003:1012 arg1 amount 1039:1044 amount 1039:1044 amount of enzyme 1039:1054 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 7 60 from dependency 1003:1012 arg1 time 1033:1036 time 1033:1036 time 1033:1036 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 11 61 theme aqueous 1759:1765 arg1 washes 1767:1772 extensive aqueous washes 1749:1772 extensive aqueous washes 1749:1772 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 7 62 theme activity 1021:1028 arg1 dependency 1003:1012 linear dependency 996:1012 linear dependency of its activity on time, amount of enzyme, and substrates 996:1070 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 12 63 theme other 1863:1867 arg1 glycosyltransferases 1869:1888 other glycosyltransferases 1863:1888 other glycosyltransferases 1863:1888 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 7 64 theme enzymatic 970:978 arg1 reaction 980:987 The enzymatic reaction 966:987 The enzymatic reaction 966:987 The enzymatic reaction showed linear dependency of its activity on time, amount of enzyme, and substrates. 9023546 1 65 theme GDP-fucose 134:143 arg1 fucosyltransferase 157:174 GDP-fucose:polypeptide fucosyltransferase 134:174 GDP-fucose:polypeptide fucosyltransferase 134:174 An assay of GDP-fucose:polypeptide fucosyltransferase has been established. 9023546 0 66 theme O-linked 90:97 arg1 fucose 99:104 O-linked fucose 90:104 O-linked fucose 90:104 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 1 67 theme polypeptide 145:155 arg1 fucosyltransferase 157:174 GDP-fucose:polypeptide fucosyltransferase 134:174 GDP-fucose:polypeptide fucosyltransferase 134:174 An assay of GDP-fucose:polypeptide fucosyltransferase has been established. 9023546 11 68 theme X-100 1700:1704 arg1 extraction 1706:1715 Triton X-100 extraction 1693:1715 Triton X-100 extraction of the membrane particles 1693:1741 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 4 69 dep contain 537:543 arg1 Spellman 573:580 Spellman 573:580 Spellman 573:580 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 4 69 dep contain 537:543 arg1 Harris 562:567 Harris 562:567 Harris 562:567 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 2 70 from residue 320:326 arg1 domains 335:341 EGF domains 331:341 EGF domains 331:341 The enzyme catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains. 9023546 3 71 theme human 371:375 arg1 VII 384:386 recombinant human factor VII 359:386 recombinant human factor VII EGF-1 domain 359:399 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 4 72 with peptides 468:475 arg1 sequences 482:490 sequences 482:490 sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) 482:613 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 5 73 theme EGF 790:792 arg1 structure 794:802 the EGF structure 786:802 the EGF structure 786:802 These synthetic peptides did not compromise complete EGF domains and did not contain all six cysteine residues that define the EGF structure. 9023546 4 74 dep Harris 562:567 arg1 Glycobiology 589:600 Glycobiology 589:600 Glycobiology 589:600 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 12 75 theme membrane 1825:1832 arg1 protein 1834:1840 a membrane protein 1823:1840 a membrane protein 1823:1840 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 12 75 theme membrane 1825:1832 arg1 enzyme 1804:1809 the enzyme 1800:1809 the enzyme 1800:1809 The result suggests that the enzyme is probably a membrane protein and, by analogy with other glycosyltransferases, probably has a 'stem' region that is very susceptible to proteolysis. 9023546 0 76 theme GDP-L-fucose 20:31 arg1 Identification 0:13 Identification 0:13 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 0:120 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 3 77 theme VII 384:386 arg1 domain 394:399 recombinant human factor VII EGF-1 domain 359:399 recombinant human factor VII EGF-1 domain 359:399 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 9 78 dep in 1220:1221 arg1 vitro 1223:1227 vitro 1223:1227 vitro 1223:1227 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 0 79 theme polypeptide 33:43 arg1 fucosyltransferase 45:62 polypeptide fucosyltransferase 33:62 polypeptide fucosyltransferase 33:62 Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains. 9023546 9 80 theme substrate 1300:1308 arg1 conversion 1273:1282 complete conversion 1264:1282 complete conversion of the acceptor substrate to glycosylated product 1264:1332 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 4 81 theme O-linked 545:552 arg1 fucose 554:559 O-linked fucose 545:559 O-linked fucose 545:559 Synthetic peptides with sequences taken from five proteins previously shown to contain O-linked fucose (Harris and Spellman, 1993; Glycobiology, 3, 219-224) did not serve as efficient acceptor substrates. 9023546 9 82 theme glycosylated 1313:1324 arg1 product 1326:1332 glycosylated product 1313:1332 glycosylated product 1313:1332 The in vitro glycosylation reaction resulted in complete conversion of the acceptor substrate to glycosylated product, and characterization of the purified product by electrospray mass spectrometry revealed that one fucose was added onto the polypeptide. 9023546 3 83 theme acceptor 404:411 arg1 substrate 413:421 acceptor substrate 404:421 acceptor substrate 404:421 The assay uses recombinant human factor VII EGF-1 domain as acceptor substrate and GDP-fucose as donor substrate. 9023546 11 84 theme inhibitors 1644:1653 arg1 presence 1623:1630 the presence 1619:1630 the presence of protease inhibitors 1619:1653 However, when enzyme was prepared from rat liver in the presence of protease inhibitors, 37% of the activity was recovered by Triton X-100 extraction of the membrane particles after extensive aqueous washes. 9023546 2 85 theme EGF 331:333 arg1 domains 335:341 EGF domains 331:341 EGF domains 331:341 The enzyme catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue in EGF domains. 20581009 6 0 theme B 1238:1238 arg1 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 0 theme B 1238:1238 arg1 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 0 theme B 1238:1238 arg1 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 0 theme B 1238:1238 arg1 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 3 1 theme over-expression 690:704 arg1 system 706:711 a recombinant over-expression system 676:711 a recombinant over-expression system in human embryonic kidney cells (HEK) 676:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 4 2 theme GAG 871:873 arg1 chains 875:880 the GAG chains 867:880 the GAG chains from both sources 867:898 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 1 3 theme soluble 163:169 arg1 CS/DS 201:205 CS/DS 201:205 CS/DS 201:205 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 1 3 theme soluble 163:169 arg1 sulfate 192:198 soluble chondroitin/dermatan sulfate 163:198 a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan 141:219 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 5 4 dep DS-specific 1019:1029 arg1 IdoA-containing 1032:1046 IdoA-containing 1032:1046 IdoA-containing 1032:1046 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 6 5 theme disaccharide 1240:1251 arg1 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 5 theme disaccharide 1240:1251 arg1 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 5 theme disaccharide 1240:1251 arg1 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 5 theme disaccharide 1240:1251 arg1 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 5 6 theme % 1093:1093 arg1 2-O-sulfated 1108:1119 2-O-sulfated 1108:1119 2-O-sulfated 1108:1119 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 5 7 theme DS-specific 1019:1029 arg1 disaccharides 1048:1060 The DS-specific, IdoA-containing disaccharides 1015:1060 The DS-specific, IdoA-containing disaccharides 1015:1060 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 1 8 theme chondroitin/dermatan 171:190 arg1 CS/DS 201:205 CS/DS 201:205 CS/DS 201:205 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 1 8 theme chondroitin/dermatan 171:190 arg1 sulfate 192:198 soluble chondroitin/dermatan sulfate 163:198 a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan 141:219 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 0 9 theme soluble 96:102 arg1 Characterization 0:15 Characterization 0:15 Characterization 0:15 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 0 9 theme soluble 96:102 arg1 proteoglycan 116:127 a soluble endothelial proteoglycan 94:127 a soluble endothelial proteoglycan 94:127 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 7 10 theme possible 1665:1672 arg1 model 1674:1678 a possible model 1663:1678 a possible model of Endocan CS/DS chain organization 1663:1714 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 4 11 theme size 950:953 arg1 exception 937:945 the exception 933:945 the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells) 933:1012 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 6 12 theme E 1231:1231 arg1 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 12 theme E 1231:1231 arg1 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 12 theme E 1231:1231 arg1 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 12 theme E 1231:1231 arg1 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 3 13 theme human 716:720 arg1 HEK 746:748 HEK 746:748 HEK 746:748 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 13 theme human 716:720 arg1 cells 739:743 human embryonic kidney cells 716:743 human embryonic kidney cells (HEK) 716:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 4 14 theme HEK-293 999:1005 arg1 cells 1007:1011 HEK-293 cells 999:1011 HEK-293 cells 999:1011 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 7 15 theme CS/DS 1548:1552 arg1 chains 1554:1559 authentic CS/DS chains 1538:1559 authentic CS/DS chains 1538:1559 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 3 16 theme embryonic 722:730 arg1 HEK 746:748 HEK 746:748 HEK 746:748 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 16 theme embryonic 722:730 arg1 cells 739:743 human embryonic kidney cells 716:743 human embryonic kidney cells (HEK) 716:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 2 17 dep factor 370:375 arg1 activities 387:396 mitogenic activities 377:396 mitogenic activities 377:396 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 6 18 theme HGF/SF-binding 1396:1409 arg1 activity 1411:1418 the HGF/SF-binding activity 1392:1418 the HGF/SF-binding activity of the CS/DS 1392:1431 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 1 19 theme sulfate 192:198 arg1 Endocan 130:136 Endocan 130:136 Endocan 130:136 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 1 19 theme sulfate 192:198 arg1 proteoglycan 208:219 a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan 141:219 a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan 141:219 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 0 20 theme endothelial 104:114 arg1 Characterization 0:15 Characterization 0:15 Characterization 0:15 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 0 20 theme endothelial 104:114 arg1 proteoglycan 116:127 a soluble endothelial proteoglycan 94:127 a soluble endothelial proteoglycan 94:127 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 4 21 theme resonance 835:843 arg1 studies 845:851 nuclear magnetic resonance studies 818:851 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies 752:851 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 7 22 theme binding 1632:1638 arg1 analysis 1640:1647 structural and/or binding analysis 1614:1647 structural and/or binding analysis 1614:1647 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 3 23 theme glycosaminoglycan 534:550 arg1 chain 558:562 the glycosaminoglycan (GAG) chain 530:562 the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK) 530:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 24 from system 706:711 arg1 HEK 746:748 HEK 746:748 HEK 746:748 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 24 from system 706:711 arg1 cells 739:743 human embryonic kidney cells 716:743 human embryonic kidney cells (HEK) 716:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 5 25 theme chain 1084:1088 arg1 % 1075:1075 30% 1073:1075 30% of the chain (15% of which are 2-O-sulfated) 1073:1120 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 5 25 theme chain 1084:1088 arg1 chain 1084:1088 the chain 1080:1088 the chain (15% of which are 2-O-sulfated) 1080:1120 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 6 26 dep units 1253:1257 arg1 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 26 dep units 1253:1257 arg1 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 26 dep units 1253:1257 arg1 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 26 dep units 1253:1257 arg1 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 7 27 theme authentic 1538:1546 arg1 chains 1554:1559 authentic CS/DS chains 1538:1559 authentic CS/DS chains 1538:1559 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 7 28 dep provide 1496:1502 arg1 providing 1653:1661 providing 1653:1661 providing a possible model of Endocan CS/DS chain organization 1653:1714 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 7 28 dep provide 1496:1502 arg1 enabling 1562:1569 enabling 1562:1569 enabling the purification of sufficient amounts for structural and/or binding analysis 1562:1647 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 7 29 theme sufficient 1591:1600 arg1 amounts 1602:1608 sufficient amounts 1591:1608 sufficient amounts for structural and/or binding analysis 1591:1647 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 7 30 theme structural 1614:1623 arg1 analysis 1640:1647 structural and/or binding analysis 1614:1647 structural and/or binding analysis 1614:1647 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 3 31 theme Endocan 567:573 arg1 chain 558:562 the glycosaminoglycan (GAG) chain 530:562 the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK) 530:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 7 32 theme CS/DS 1691:1695 arg1 organization 1703:1714 Endocan CS/DS chain organization 1683:1714 Endocan CS/DS chain organization 1683:1714 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 2 33 theme mitogenic 377:385 arg1 activities 387:396 mitogenic activities 377:396 mitogenic activities 377:396 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 6 34 located detected 1325:1332 arg2 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 34 located detected 1325:1332 arg1 amounts 1349:1355 significant amounts 1337:1355 significant amounts in both chains 1337:1370 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 34 located detected 1325:1332 arg2 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 34 located detected 1325:1332 arg2 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 34 located detected 1325:1332 arg2 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 4 35 theme nuclear 818:824 arg1 resonance 835:843 nuclear magnetic resonance 818:843 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies 752:851 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 7 36 theme Endocan 1525:1531 arg1 source 1515:1520 a valuable source 1504:1520 a valuable source of Endocan with authentic CS/DS chains 1504:1559 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 0 37 theme binding 21:27 arg1 activity 29:36 binding activity 21:36 binding activity 21:36 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 3 38 theme kidney 732:737 arg1 HEK 746:748 HEK 746:748 HEK 746:748 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 38 theme kidney 732:737 arg1 cells 739:743 human embryonic kidney cells 716:743 human embryonic kidney cells (HEK) 716:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 4 39 dep size 950:953 arg1 40 kDa 963:968 40 kDa 963:968 40 kDa 963:968 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 4 39 dep size 950:953 arg1 15 956:957 15 956:957 15 956:957 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 3 40 theme endothelial 642:652 arg1 HUVEC 661:665 HUVEC 661:665 HUVEC 661:665 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 40 theme endothelial 642:652 arg1 cells 654:658 the naturally producing human umbilical vein endothelial cells 597:658 the naturally producing human umbilical vein endothelial cells (HUVEC) 597:666 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 7 41 theme valuable 1506:1513 arg1 source 1515:1520 a valuable source 1504:1520 a valuable source of Endocan with authentic CS/DS chains 1504:1559 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 2 42 theme cellular 475:482 arg1 proliferation 484:496 cellular proliferation 475:496 cellular proliferation 475:496 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 0 43 from Endocan 85:91 arg1 Characterization 0:15 Characterization 0:15 Characterization 0:15 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 0 43 from Endocan 85:91 arg1 activity 29:36 binding activity 21:36 binding activity 21:36 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 0 43 from Endocan 85:91 arg1 proteoglycan 116:127 a soluble endothelial proteoglycan 94:127 a soluble endothelial proteoglycan 94:127 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 6 44 from amounts 1349:1355 arg1 chains 1365:1370 both chains 1360:1370 both chains 1360:1370 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 5 45 theme saccharide 1192:1201 arg1 domains 1203:1209 saccharide domains 1192:1209 saccharide domains 1192:1209 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 2 46 theme factor/scatter 417:430 arg1 HGF/SF 440:445 HGF/SF 440:445 HGF/SF 440:445 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 46 theme factor/scatter 417:430 arg1 factor 432:437 hepatocyte growth factor/scatter factor 399:437 hepatocyte growth factor/scatter factor (HGF/SF) 399:446 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 46 theme factor/scatter 417:430 arg1 factor 370:375 growth factor 363:375 growth factor 363:375 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 47 theme hepatocyte 399:408 arg1 HGF/SF 440:445 HGF/SF 440:445 HGF/SF 440:445 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 47 theme hepatocyte 399:408 arg1 factor 432:437 hepatocyte growth factor/scatter factor 399:437 hepatocyte growth factor/scatter factor (HGF/SF) 399:446 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 47 theme hepatocyte 399:408 arg1 factor 370:375 growth factor 363:375 growth factor 363:375 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 3 48 theme umbilical 627:635 arg1 HUVEC 661:665 HUVEC 661:665 HUVEC 661:665 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 48 theme umbilical 627:635 arg1 cells 654:658 the naturally producing human umbilical vein endothelial cells 597:658 the naturally producing human umbilical vein endothelial cells (HUVEC) 597:666 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 2 49 theme growth 410:415 arg1 HGF/SF 440:445 HGF/SF 440:445 HGF/SF 440:445 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 49 theme growth 410:415 arg1 factor 432:437 hepatocyte growth factor/scatter factor 399:437 hepatocyte growth factor/scatter factor (HGF/SF) 399:446 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 49 theme growth 410:415 arg1 factor 370:375 growth factor 363:375 growth factor 363:375 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 0 50 theme sulfate 66:72 arg1 chain 74:78 the chondroitin/dermatan sulfate chain 41:78 the chondroitin/dermatan sulfate chain 41:78 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 5 51 dep chain 1084:1088 arg1 % 1093:1093 15% 1091:1093 15% of which are 2-O-sulfated 1091:1119 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 7 52 with source 1515:1520 arg1 chains 1554:1559 authentic CS/DS chains 1538:1559 authentic CS/DS chains 1538:1559 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 3 53 theme vein 637:640 arg1 HUVEC 661:665 HUVEC 661:665 HUVEC 661:665 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 53 theme vein 637:640 arg1 cells 654:658 the naturally producing human umbilical vein endothelial cells 597:658 the naturally producing human umbilical vein endothelial cells (HUVEC) 597:666 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 4 54 theme magnetic 826:833 arg1 resonance 835:843 nuclear magnetic resonance 818:843 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies 752:851 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 0 55 theme chondroitin/dermatan 45:64 arg1 chain 74:78 the chondroitin/dermatan sulfate chain 41:78 the chondroitin/dermatan sulfate chain 41:78 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 7 56 theme HEK-293 1461:1467 arg1 cells 1469:1473 HEK-293 cells 1461:1473 HEK-293 cells 1461:1473 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 6 57 theme sulfated 1219:1226 arg1 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 57 theme sulfated 1219:1226 arg1 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 57 theme sulfated 1219:1226 arg1 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 57 theme sulfated 1219:1226 arg1 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 2 58 theme endothelial 237:247 arg1 cells 249:253 endothelial cells 237:253 endothelial cells 237:253 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 7 59 theme chain 1697:1701 arg1 organization 1703:1714 Endocan CS/DS chain organization 1683:1714 Endocan CS/DS chain organization 1683:1714 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 7 60 theme amounts 1602:1608 arg1 purification 1575:1586 the purification 1571:1586 the purification of sufficient amounts for structural and/or binding analysis 1571:1647 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 4 61 theme many 906:909 arg1 characteristics 911:925 many characteristics 906:925 many characteristics 906:925 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 4 62 theme different 781:789 arg1 chondroitinases 791:805 different chondroitinases 781:805 different chondroitinases 781:805 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 3 63 theme producing 611:619 arg1 HUVEC 661:665 HUVEC 661:665 HUVEC 661:665 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 63 theme producing 611:619 arg1 cells 654:658 the naturally producing human umbilical vein endothelial cells 597:658 the naturally producing human umbilical vein endothelial cells (HUVEC) 597:666 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 7 64 theme organization 1703:1714 arg1 model 1674:1678 a possible model 1663:1678 a possible model of Endocan CS/DS chain organization 1663:1714 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 2 65 theme tumors 338:343 arg1 number 328:333 a number 326:333 a number of tumors 326:343 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 4 66 theme Compositional 752:764 arg1 analysis 766:773 Compositional analysis 752:773 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies 752:851 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 4 67 from sources 892:898 arg1 chains 875:880 the GAG chains 867:880 the GAG chains from both sources 867:898 Compositional analysis using different chondroitinases as well as nuclear magnetic resonance studies revealed that the GAG chains from both sources share many characteristics, with the exception of size (15 and 40 kDa, respectively, for HUVEC and HEK-293 cells). 20581009 3 68 theme human 621:625 arg1 HUVEC 661:665 HUVEC 661:665 HUVEC 661:665 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 3 68 theme human 621:625 arg1 cells 654:658 the naturally producing human umbilical vein endothelial cells 597:658 the naturally producing human umbilical vein endothelial cells (HUVEC) 597:666 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 7 69 theme Endocan 1683:1689 arg1 organization 1703:1714 Endocan CS/DS chain organization 1683:1714 Endocan CS/DS chain organization 1683:1714 This work establishes that HEK-293 cells can be engineered to provide a valuable source of Endocan with authentic CS/DS chains, enabling the purification of sufficient amounts for structural and/or binding analysis and providing a possible model of Endocan CS/DS chain organization. 20581009 1 70 theme identified 152:161 arg1 Endocan 130:136 Endocan 130:136 Endocan 130:136 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 1 70 theme identified 152:161 arg1 proteoglycan 208:219 a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan 141:219 a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan 141:219 Endocan is a recently identified soluble chondroitin/dermatan sulfate (CS/DS) proteoglycan. 20581009 6 71 theme D 1228:1228 arg1 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 HexA-GalNAc4S6S 1277:1291 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 71 theme D 1228:1228 arg1 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 HexA2S-GalNAc4S 1298:1312 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 71 theme D 1228:1228 arg1 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 HexA2S-GalNAc6S 1260:1274 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 71 theme D 1228:1228 arg1 units 1253:1257 Highly sulfated D, E, and B disaccharide units 1212:1257 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) 1212:1313 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 5 72 dep tetra- 1150:1155 arg1 domains 1203:1209 saccharide domains 1192:1209 saccharide domains 1192:1209 The DS-specific, IdoA-containing disaccharides contribute 30% of the chain (15% of which are 2-O-sulfated) and are mostly clustered in tetra- (35%), hexa- (12%), and octa- (5%) saccharide domains. 20581009 6 73 theme CS/DS 1427:1431 arg1 activity 1411:1418 the HGF/SF-binding activity 1392:1418 the HGF/SF-binding activity of the CS/DS 1392:1431 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 6 74 theme significant 1337:1347 arg1 amounts 1349:1355 significant amounts 1337:1355 significant amounts in both chains 1337:1370 Highly sulfated D, E, and B disaccharide units (HexA2S-GalNAc6S, HexA-GalNAc4S6S, and HexA2S-GalNAc4S) were also detected in significant amounts in both chains and may account for the HGF/SF-binding activity of the CS/DS. 20581009 0 75 theme chain 74:78 arg1 Characterization 0:15 Characterization 0:15 Characterization 0:15 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 0 75 theme chain 74:78 arg1 activity 29:36 binding activity 21:36 binding activity 21:36 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 0 75 theme chain 74:78 arg1 proteoglycan 116:127 a soluble endothelial proteoglycan 94:127 a soluble endothelial proteoglycan 94:127 Characterization and binding activity of the chondroitin/dermatan sulfate chain from Endocan, a soluble endothelial proteoglycan. 20581009 3 76 theme recombinant 678:688 arg1 system 706:711 a recombinant over-expression system 676:711 a recombinant over-expression system in human embryonic kidney cells (HEK) 676:749 In this work, we characterized the glycosaminoglycan (GAG) chain of Endocan, purified either from the naturally producing human umbilical vein endothelial cells (HUVEC) or from a recombinant over-expression system in human embryonic kidney cells (HEK). 20581009 2 77 theme growth 363:368 arg1 factor 432:437 hepatocyte growth factor/scatter factor 399:437 hepatocyte growth factor/scatter factor (HGF/SF) 399:446 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 20581009 2 77 theme growth 363:368 arg1 factor 370:375 growth factor 363:375 growth factor 363:375 Synthesized by endothelial cells, it has been found to be over-expressed in the vasculature surrounding a number of tumors, and by promoting growth factor mitogenic activities, hepatocyte growth factor/scatter factor (HGF/SF) in particular, it supports cellular proliferation. 15010458 0 0 theme Drosophila 46:55 arg1 locus 68:72 Drosophila flightless locus 46:72 Drosophila flightless locus 46:72 A novel human Cl(-) channel family related to Drosophila flightless locus. 15010458 2 1 theme human 393:397 arg1 homologues 399:408 human homologues 393:408 human homologues of tweety (hTTYH1-3) 393:429 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 1 theme human 393:397 arg1 channels 452:459 novel maxi-Cl(-) channels 435:459 novel maxi-Cl(-) channels 435:459 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 6 2 theme 2'-disulfonic 922:934 arg1 sensitivity 941:951 2'-disulfonic acid sensitivity 922:951 2'-disulfonic acid sensitivity 922:951 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 1 3 theme little 177:182 arg1 information 184:194 little information 177:194 little information on the molecular nature of the channel underlying this conductance 177:261 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 7 4 theme ionomycin-induced 1051:1067 arg1 maxi-Cl 1069:1075 an ionomycin-induced maxi-Cl 1048:1075 an ionomycin-induced maxi-Cl(-) channel 1048:1086 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 7 4 theme ionomycin-induced 1051:1067 arg1 hTTYH2 1033:1038 hTTYH2 1033:1038 hTTYH2 1033:1038 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 6 5 theme current 854:860 arg1 voltage 862:868 26-picosiemen linear current voltage 833:868 26-picosiemen linear current voltage 833:868 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 8 6 theme hTTYH 1186:1190 arg1 family 1192:1197 the hTTYH family 1182:1197 the hTTYH family 1182:1197 Therefore, the hTTYH family encoded maxi-Cl(-) channels of mammals. 15010458 8 6 theme hTTYH 1186:1190 arg1 channels 1218:1225 maxi-Cl(-) channels 1207:1225 maxi-Cl(-) channels of mammals 1207:1236 Therefore, the hTTYH family encoded maxi-Cl(-) channels of mammals. 15010458 6 7 theme Cl 1015:1016 arg1 4,4'-diisothiocyanato-stilbene-2 889:920 4,4'-diisothiocyanato-stilbene-2 889:920 4,4'-diisothiocyanato-stilbene-2 889:920 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 7 theme Cl 1015:1016 arg1 voltage 862:868 26-picosiemen linear current voltage 833:868 26-picosiemen linear current voltage 833:868 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 7 theme Cl 1015:1016 arg1 order 991:995 the permeability order 974:995 the permeability order of I(-) > Br(-) > Cl(-) 974:1019 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 7 theme Cl 1015:1016 arg1 kinetics 879:886 complex kinetics 871:886 complex kinetics 871:886 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 7 theme Cl 1015:1016 arg1 sensitivity 941:951 2'-disulfonic acid sensitivity 922:951 2'-disulfonic acid sensitivity 922:951 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 7 theme Cl 1015:1016 arg1 subconductance 954:967 subconductance 954:967 subconductance 954:967 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 8 theme linear 847:852 arg1 voltage 862:868 26-picosiemen linear current voltage 833:868 26-picosiemen linear current voltage 833:868 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 0 9 theme flightless 57:66 arg1 locus 68:72 Drosophila flightless locus 46:72 Drosophila flightless locus 46:72 A novel human Cl(-) channel family related to Drosophila flightless locus. 15010458 6 10 theme maxi-Cl 772:778 arg1 channel 783:789 a maxi-Cl(-) channel 770:789 a maxi-Cl(-) channel 770:789 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 11 theme > 1013:1013 arg1 Cl 1015:1016 I(-) > Br(-) > Cl 1000:1016 I(-) > Br(-) > Cl 1000:1016 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 6 12 theme 26-picosiemen 833:845 arg1 voltage 862:868 26-picosiemen linear current voltage 833:868 26-picosiemen linear current voltage 833:868 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 5 13 theme pore 672:675 arg1 mutants 677:683 pore mutants 672:683 pore mutants 672:683 Analysis of pore mutants suggested that positively charged amino acids contributed to anion selectivity. 15010458 5 14 theme anion 746:750 arg1 selectivity 752:762 anion selectivity 746:762 anion selectivity 746:762 Analysis of pore mutants suggested that positively charged amino acids contributed to anion selectivity. 15010458 2 15 theme Drosophila 310:319 arg1 flightless 321:330 Drosophila flightless 310:330 Drosophila flightless 310:330 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 16 theme tweety 413:418 arg1 homologues 399:408 human homologues 393:408 human homologues of tweety (hTTYH1-3) 393:429 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 16 theme tweety 413:418 arg1 channels 452:459 novel maxi-Cl(-) channels 435:459 novel maxi-Cl(-) channels 435:459 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 1 17 dep chloride 93:100 arg1 maxi-Cl 103:109 maxi-Cl 103:109 maxi-Cl 103:109 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 4 18 theme whole 528:532 arg1 large 561:565 large 561:565 large 561:565 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 15010458 4 18 theme whole 528:532 arg1 current 539:545 The whole cell current 524:545 The whole cell current of hTTYH3 524:555 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 15010458 0 19 theme novel 2:6 arg1 channel 20:26 A novel human Cl(-) channel 0:26 A novel human Cl(-) channel family 0:33 A novel human Cl(-) channel family related to Drosophila flightless locus. 15010458 8 20 theme maxi-Cl 1207:1213 arg1 family 1192:1197 the hTTYH family 1182:1197 the hTTYH family 1182:1197 Therefore, the hTTYH family encoded maxi-Cl(-) channels of mammals. 15010458 8 20 theme maxi-Cl 1207:1213 arg1 channels 1218:1225 maxi-Cl(-) channels 1207:1225 maxi-Cl(-) channels of mammals 1207:1236 Therefore, the hTTYH family encoded maxi-Cl(-) channels of mammals. 15010458 2 21 theme located 299:305 arg1 tweety 284:289 tweety 284:289 tweety 284:289 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 21 theme located 299:305 arg1 gene 294:297 a gene 292:297 a gene located in Drosophila flightless 292:330 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 5 22 theme mutants 677:683 arg1 Analysis 660:667 Analysis 660:667 Analysis of pore mutants 660:683 Analysis of pore mutants suggested that positively charged amino acids contributed to anion selectivity. 15010458 1 23 theme molecular 203:211 arg1 nature 213:218 the molecular nature 199:218 the molecular nature of the channel underlying this conductance 199:261 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 9 24 theme channel 1367:1373 arg1 molecules 1375:1383 novel Cl(-) channel molecules 1355:1383 novel Cl(-) channel molecules 1355:1383 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 9 25 from studies 1247:1253 arg1 family 1268:1273 the hTTYH family 1258:1273 the hTTYH family 1258:1273 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 6 26 theme Br 1007:1008 arg1 Cl 1015:1016 I(-) > Br(-) > Cl 1000:1016 I(-) > Br(-) > Cl 1000:1016 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 2 27 theme similar 349:355 arg1 structure 339:347 a structure 337:347 a structure similar to those of known channels 337:382 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 4 28 with treatment 634:642 arg1 ionomycin 649:657 ionomycin 649:657 ionomycin 649:657 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 15010458 7 29 dep maxi-Cl 1069:1075 arg1 channel 1080:1086 channel 1080:1086 channel 1080:1086 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 6 30 theme acid 936:939 arg1 sensitivity 941:951 2'-disulfonic acid sensitivity 922:951 2'-disulfonic acid sensitivity 922:951 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 0 31 theme human 8:12 arg1 Cl 14:15 human Cl 8:15 human Cl 8:15 A novel human Cl(-) channel family related to Drosophila flightless locus. 15010458 6 32 theme single 811:816 arg1 channel 818:824 the hTTYH3 single channel 800:824 the hTTYH3 single channel 800:824 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 2 33 theme maxi-Cl 441:447 arg1 homologues 399:408 human homologues 393:408 human homologues of tweety (hTTYH1-3) 393:429 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 33 theme maxi-Cl 441:447 arg1 channels 452:459 novel maxi-Cl(-) channels 435:459 novel maxi-Cl(-) channels 435:459 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 1 34 theme Large 75:79 arg1 chloride 93:100 Large conductance chloride 75:100 Large conductance chloride (maxi-Cl(-)) currents 75:122 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 5 35 theme charged 711:717 arg1 acids 725:729 positively charged amino acids 700:729 positively charged amino acids 700:729 Analysis of pore mutants suggested that positively charged amino acids contributed to anion selectivity. 15010458 6 36 theme hTTYH3 804:809 arg1 channel 818:824 the hTTYH3 single channel 800:824 the hTTYH3 single channel 800:824 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 2 37 theme novel 435:439 arg1 maxi-Cl 441:447 novel maxi-Cl 435:447 novel maxi-Cl 435:447 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 9 38 theme novel 1355:1359 arg1 Cl 1361:1362 novel Cl 1355:1362 novel Cl 1355:1362 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 5 39 theme amino 719:723 arg1 acids 725:729 positively charged amino acids 700:729 positively charged amino acids 700:729 Analysis of pore mutants suggested that positively charged amino acids contributed to anion selectivity. 15010458 4 40 theme cell 534:537 arg1 large 561:565 large 561:565 large 561:565 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 15010458 4 40 theme cell 534:537 arg1 current 539:545 The whole cell current 524:545 The whole cell current of hTTYH3 524:555 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 15010458 1 41 theme channel 227:233 arg1 nature 213:218 the molecular nature 199:218 the molecular nature of the channel underlying this conductance 199:261 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 1 42 from information 184:194 arg1 nature 213:218 the molecular nature 199:218 the molecular nature of the channel underlying this conductance 199:261 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 3 43 theme excitable 505:513 arg1 tissues 515:521 excitable tissues 505:521 excitable tissues 505:521 hTTYH3 mRNA was found to be distributed in excitable tissues. 15010458 0 44 theme Cl 14:15 arg1 channel 20:26 A novel human Cl(-) channel 0:26 A novel human Cl(-) channel family 0:33 A novel human Cl(-) channel family related to Drosophila flightless locus. 15010458 2 45 from located 299:305 arg1 flightless 321:330 Drosophila flightless 310:330 Drosophila flightless 310:330 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 9 46 theme Further 1239:1245 arg1 studies 1247:1253 Further studies 1239:1253 Further studies on the hTTYH family 1239:1273 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 0 47 theme channel 20:26 arg1 family 28:33 A novel human Cl(-) channel family 0:33 A novel human Cl(-) channel family 0:33 A novel human Cl(-) channel family related to Drosophila flightless locus. 15010458 3 48 theme hTTYH3 462:467 arg1 mRNA 469:472 hTTYH3 mRNA 462:472 hTTYH3 mRNA 462:472 hTTYH3 mRNA was found to be distributed in excitable tissues. 15010458 1 49 theme conductance 81:91 arg1 chloride 93:100 Large conductance chloride 75:100 Large conductance chloride (maxi-Cl(-)) currents 75:122 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 9 50 theme hTTYH 1262:1266 arg1 family 1268:1273 the hTTYH family 1258:1273 the hTTYH family 1258:1273 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 8 51 theme mammals 1230:1236 arg1 family 1192:1197 the hTTYH family 1182:1197 the hTTYH family 1182:1197 Therefore, the hTTYH family encoded maxi-Cl(-) channels of mammals. 15010458 8 51 theme mammals 1230:1236 arg1 channels 1218:1225 maxi-Cl(-) channels 1207:1225 maxi-Cl(-) channels of mammals 1207:1236 Therefore, the hTTYH family encoded maxi-Cl(-) channels of mammals. 15010458 9 52 theme molecules 1375:1383 arg1 roles 1346:1350 physiological and pathophysiological roles 1309:1350 physiological and pathophysiological roles of novel Cl(-) channel molecules 1309:1383 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 1 53 theme chloride 93:100 arg1 currents 115:122 Large conductance chloride (maxi-Cl(-)) currents 75:122 Large conductance chloride (maxi-Cl(-)) currents 75:122 Large conductance chloride (maxi-Cl(-)) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. 15010458 7 54 theme swelling-activated 1132:1149 arg1 maxi-Cl 1151:1157 swelling-activated maxi-Cl 1132:1157 swelling-activated maxi-Cl 1132:1157 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 9 55 theme Cl 1361:1362 arg1 molecules 1375:1383 novel Cl(-) channel molecules 1355:1383 novel Cl(-) channel molecules 1355:1383 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 6 56 theme I 1000:1000 arg1 Cl 1015:1016 I(-) > Br(-) > Cl 1000:1016 I(-) > Br(-) > Cl 1000:1016 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 9 57 theme roles 1346:1350 arg1 elucidation 1294:1304 the elucidation 1290:1304 the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules 1290:1383 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 9 58 theme physiological 1309:1321 arg1 roles 1346:1350 physiological and pathophysiological roles 1309:1350 physiological and pathophysiological roles of novel Cl(-) channel molecules 1309:1383 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 2 59 contain has 333:335 arg1 tweety 284:289 tweety 284:289 tweety 284:289 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 59 contain has 333:335 arg1 gene 294:297 a gene 292:297 a gene located in Drosophila flightless 292:330 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 2 59 contain has 333:335 arg2 structure 339:347 a structure 337:347 a structure similar to those of known channels 337:382 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 7 60 theme maxi-Cl 1151:1157 arg1 TTYH1 1093:1097 TTYH1 1093:1097 TTYH1 1093:1097 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 7 60 theme maxi-Cl 1151:1157 arg1 channel 1162:1168 a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel 1107:1168 a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel 1107:1168 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 6 61 theme permeability 978:989 arg1 order 991:995 the permeability order 974:995 the permeability order of I(-) > Br(-) > Cl(-) 974:1019 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 7 62 theme -independent 1115:1126 arg1 TTYH1 1093:1097 TTYH1 1093:1097 TTYH1 1093:1097 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 7 62 theme -independent 1115:1126 arg1 channel 1162:1168 a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel 1107:1168 a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel 1107:1168 Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl(-) channel, but TTYH1 encoded a Ca(2+)-independent and swelling-activated maxi-Cl(-) channel. 15010458 2 63 from flightless 321:330 arg1 located 299:305 located 299:305 located 299:305 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 9 64 theme pathophysiological 1327:1344 arg1 roles 1346:1350 physiological and pathophysiological roles 1309:1350 physiological and pathophysiological roles of novel Cl(-) channel molecules 1309:1383 Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl(-) channel molecules. 15010458 6 65 theme complex 871:877 arg1 kinetics 879:886 complex kinetics 871:886 complex kinetics 871:886 Like a maxi-Cl(-) channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I(-) > Br(-) > Cl(-). 15010458 2 66 theme known 369:373 arg1 channels 375:382 known channels 369:382 known channels 369:382 We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1-3) are novel maxi-Cl(-) channels. 15010458 4 67 theme hTTYH3 550:555 arg1 large 561:565 large 561:565 large 561:565 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 15010458 4 67 theme hTTYH3 550:555 arg1 current 539:545 The whole cell current 524:545 The whole cell current of hTTYH3 524:555 The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. 18508581 0 0 theme antigen 70:76 arg1 EpCAM 78:82 cancer stem cell antigen EpCAM 53:82 cancer stem cell antigen EpCAM 53:82 Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. 18508581 3 1 gly glycosylation 497:509 arg1 EpCAM 514:518 EpCAM 514:518 EpCAM 514:518 We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. 18508581 7 2 theme EpCAM 880:884 arg1 Mutants 869:875 Mutants 869:875 Mutants of EpCAM that substitute asparagine198 for alanine 869:926 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 1 3 theme increased 234:242 arg1 expression 244:253 increased expression 234:253 increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype 234:355 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 0 4 theme cell 65:68 arg1 EpCAM 78:82 cancer stem cell antigen EpCAM 53:82 cancer stem cell antigen EpCAM 53:82 Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. 18508581 7 5 theme decreased 937:945 arg1 expression 955:964 a decreased overall expression 935:964 a decreased overall expression 935:964 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 8 6 theme considerable 1031:1042 arg1 importance 1044:1053 considerable importance 1031:1053 considerable importance 1031:1053 This is of considerable importance with respect to EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality. 18508581 0 7 theme EpCAM 78:82 arg1 stability 29:37 stability 29:37 stability of tumour and cancer stem cell antigen EpCAM 29:82 Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. 18508581 7 8 theme overall 947:953 arg1 expression 955:964 a decreased overall expression 935:964 a decreased overall expression 935:964 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 3 9 theme EpCAM 514:518 arg1 means 525:529 a means 523:529 a means to discriminate normal from malignant tissues 523:575 We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. 18508581 3 9 theme EpCAM 514:518 arg1 glycosylation 497:509 differential glycosylation 484:509 differential glycosylation of EpCAM 484:518 We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. 18508581 1 10 theme target 258:263 arg1 cyclins 286:292 cyclins 286:292 cyclins 286:292 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 1 10 theme target 258:263 arg1 genes 265:269 target genes 258:269 target genes such as c-Myc 258:283 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 1 10 theme target 258:263 arg1 c-Myc 279:283 c-Myc 279:283 c-Myc 279:283 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 1 10 theme target 258:263 arg1 others 298:303 others 298:303 others 298:303 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 1 11 theme oncogenic 337:345 arg1 phenotype 347:355 an oncogenic phenotype 334:355 an oncogenic phenotype 334:355 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 4 12 gly hyperglycosylated 588:604 arg1 tissue 619:624 carcinoma tissue 609:624 carcinoma tissue 609:624 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 4 12 gly hyperglycosylated 588:604 arg1 EpCAM 578:582 EpCAM 578:582 EpCAM 578:582 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 4 12 gly hyperglycosylated 588:604 arg2 EpCAM 578:582 EpCAM 578:582 EpCAM 578:582 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 3 13 theme differential 484:495 arg1 means 525:529 a means 523:529 a means to discriminate normal from malignant tissues 523:575 We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. 18508581 3 13 theme differential 484:495 arg1 glycosylation 497:509 differential glycosylation 484:509 differential glycosylation of EpCAM 484:518 We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. 18508581 4 14 theme autologous 643:652 arg1 epithelia 661:669 autologous normal epithelia 643:669 autologous normal epithelia 643:669 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 5 15 theme extracellular 733:745 arg1 domain 747:752 EpCAM's extracellular domain 725:752 EpCAM's extracellular domain 725:752 All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. 18508581 5 16 used used 759:762 arg2 sequences 708:716 All three N-glycosylation consensus sequences 672:716 All three N-glycosylation consensus sequences within EpCAM's extracellular domain 672:752 All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. 18508581 6 17 gly glycosylation 804:816 arg1 asparagine198 821:833 asparagine198 821:833 asparagine198 821:833 We show that glycosylation at asparagine198 is crucial for protein stability. 18508581 6 17 gly glycosylation 804:816 arg2 asparagine198 821:833 asparagine198 821:833 asparagine198 821:833 We show that glycosylation at asparagine198 is crucial for protein stability. 18508581 2 18 theme epithelia 414:422 arg1 series 396:401 a series 394:401 a series of healthy epithelia 394:422 However, EpCAM is also expressed in a series of healthy epithelia, albeit generally to a far lesser extend. 18508581 1 19 dep conferring 317:326 arg1 cells 328:332 cells 328:332 cells 328:332 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 1 20 theme carcinomas 168:177 arg1 carcinomas 168:177 carcinomas 168:177 carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype 168:355 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 1 20 theme carcinomas 168:177 arg1 variety 157:163 a variety 155:163 a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype 155:355 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 2 21 theme healthy 406:412 arg1 epithelia 414:422 healthy epithelia 406:422 healthy epithelia 406:422 However, EpCAM is also expressed in a series of healthy epithelia, albeit generally to a far lesser extend. 18508581 1 22 theme genes 265:269 arg1 expression 244:253 increased expression 234:253 increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype 234:355 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 4 23 from tissue 619:624 arg1 hyperglycosylated 588:604 hyperglycosylated 588:604 hyperglycosylated 588:604 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 1 24 theme Epithelial 85:94 arg1 molecule 110:117 Epithelial cell adhesion molecule 85:117 Epithelial cell adhesion molecule EpCAM 85:123 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 5 25 theme N-glycosylation 682:696 arg1 sequences 708:716 All three N-glycosylation consensus sequences 672:716 All three N-glycosylation consensus sequences within EpCAM's extracellular domain 672:752 All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. 18508581 1 26 theme cell 96:99 arg1 molecule 110:117 Epithelial cell adhesion molecule 85:117 Epithelial cell adhesion molecule EpCAM 85:123 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 5 27 theme human 767:771 arg1 cells 784:788 human and murine cells 767:788 human and murine cells 767:788 All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. 18508581 5 28 theme consensus 698:706 arg1 sequences 708:716 All three N-glycosylation consensus sequences 672:716 All three N-glycosylation consensus sequences within EpCAM's extracellular domain 672:752 All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. 18508581 6 29 theme protein 850:856 arg1 stability 858:866 protein stability 850:866 protein stability 850:866 We show that glycosylation at asparagine198 is crucial for protein stability. 18508581 8 30 theme EpCAM 1071:1075 arg1 variants 1077:1084 EpCAM variants 1071:1084 EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality 1071:1201 This is of considerable importance with respect to EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality. 18508581 4 31 from hyperglycosylated 588:604 arg1 tissue 619:624 carcinoma tissue 609:624 carcinoma tissue 609:624 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 7 32 from membrane 1010:1017 arg1 expression 955:964 a decreased overall expression 935:964 a decreased overall expression 935:964 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 7 32 from membrane 1010:1017 arg1 half-life 970:978 half-life 970:978 half-life of the molecule at the plasma membrane 970:1017 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 3 33 theme malignant 559:567 arg1 tissues 569:575 malignant tissues 559:575 malignant tissues 559:575 We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. 18508581 6 34 from asparagine198 821:833 arg1 glycosylation 804:816 glycosylation 804:816 glycosylation at asparagine198 804:833 We show that glycosylation at asparagine198 is crucial for protein stability. 18508581 4 35 theme carcinoma 609:617 arg1 tissue 619:624 carcinoma tissue 609:624 carcinoma tissue 609:624 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 18508581 5 36 theme murine 777:782 arg1 cells 784:788 human and murine cells 767:788 human and murine cells 767:788 All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. 18508581 1 37 theme adhesion 101:108 arg1 molecule 110:117 Epithelial cell adhesion molecule 85:117 Epithelial cell adhesion molecule EpCAM 85:123 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 0 38 theme tumour 42:47 arg1 stability 29:37 stability 29:37 stability of tumour and cancer stem cell antigen EpCAM 29:82 Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. 18508581 8 39 theme normal 1099:1104 arg1 tissue 1106:1111 normal tissue 1099:1111 normal tissue 1099:1111 This is of considerable importance with respect to EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality. 18508581 1 40 theme molecule 110:117 arg1 EpCAM 119:123 Epithelial cell adhesion molecule EpCAM 85:123 Epithelial cell adhesion molecule EpCAM 85:123 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 8 41 contain have 1167:1170 arg1 it 1120:1121 it 1120:1121 it 1120:1121 This is of considerable importance with respect to EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality. 18508581 8 41 contain have 1167:1170 arg2 repercussions 1172:1184 repercussions 1172:1184 repercussions 1172:1184 This is of considerable importance with respect to EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality. 18508581 7 42 theme plasma 1003:1008 arg1 membrane 1010:1017 the plasma membrane 999:1017 the plasma membrane 999:1017 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 1 43 theme signalling 203:212 arg1 events 214:219 signalling events 203:219 signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype 203:355 Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. 18508581 2 44 dep lesser 451:456 arg1 extend 458:463 extend 458:463 extend 458:463 However, EpCAM is also expressed in a series of healthy epithelia, albeit generally to a far lesser extend. 18508581 7 45 theme molecule 987:994 arg1 expression 955:964 a decreased overall expression 935:964 a decreased overall expression 935:964 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 7 45 theme molecule 987:994 arg1 half-life 970:978 half-life 970:978 half-life of the molecule at the plasma membrane 970:1017 Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. 18508581 0 46 theme stem 60:63 arg1 EpCAM 78:82 cancer stem cell antigen EpCAM 53:82 cancer stem cell antigen EpCAM 53:82 Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. 18508581 0 47 theme cancer 53:58 arg1 EpCAM 78:82 cancer stem cell antigen EpCAM 53:82 cancer stem cell antigen EpCAM 53:82 Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. 18508581 4 48 theme normal 654:659 arg1 epithelia 661:669 autologous normal epithelia 643:669 autologous normal epithelia 643:669 EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. 16456079 1 0 theme high 153:156 arg1 affinity 158:165 high affinity 153:165 high affinity 153:165 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 4 1 theme wide 713:716 arg1 ligands 729:735 ligands 729:735 ligands 729:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 4 1 theme wide 713:716 arg1 variety 718:724 a wide variety 711:724 a wide variety of ligands 711:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 0 2 from Structure 0:8 arg1 complex 54:60 complex 54:60 complex with its receptor 54:78 Structure of human urokinase plasminogen activator in complex with its receptor. 16456079 1 3 with receptor 139:146 arg1 affinity 158:165 high affinity 153:165 high affinity 153:165 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 4 4 theme urokinase-urokinase 767:785 arg1 antagonists 796:806 urokinase-urokinase receptor antagonists 767:806 urokinase-urokinase receptor antagonists 767:806 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 2 5 theme urokinase 356:364 arg1 receptor 366:373 the urokinase receptor 352:373 the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor 352:463 We report the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor. 16456079 3 6 theme receptor 497:504 arg1 domains 476:482 The three domains 466:482 The three domains of urokinase receptor 466:504 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 3 6 theme receptor 497:504 arg1 receptor 497:504 urokinase receptor 487:504 urokinase receptor 487:504 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 2 7 from angstroms 339:347 arg1 structure 322:330 the crystal structure 310:330 the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor 310:463 We report the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor. 16456079 3 8 theme central 534:540 arg1 cavity 554:559 a central cone-shaped cavity 532:559 a central cone-shaped cavity where the urokinase fragment inserts 532:596 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 2 9 theme amino-terminal 404:417 arg1 fragment 419:426 the urokinase amino-terminal fragment 390:426 the urokinase amino-terminal fragment 390:426 We report the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor. 16456079 3 10 theme cone-shaped 542:552 arg1 cavity 554:559 a central cone-shaped cavity 532:559 a central cone-shaped cavity where the urokinase fragment inserts 532:596 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 2 11 theme urokinase 394:402 arg1 fragment 419:426 the urokinase amino-terminal fragment 390:426 the urokinase amino-terminal fragment 390:426 We report the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor. 16456079 1 12 theme signaling 181:189 arg1 cascades 191:198 signaling cascades 181:198 signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation 181:297 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 2 13 theme crystal 314:320 arg1 structure 322:330 the crystal structure 310:330 the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor 310:463 We report the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor. 16456079 0 14 theme urokinase 19:27 arg1 activator 41:49 human urokinase plasminogen activator 13:49 human urokinase plasminogen activator 13:49 Structure of human urokinase plasminogen activator in complex with its receptor. 16456079 1 15 theme plasminogen 95:105 arg1 activator 107:115 The urokinase plasminogen activator 81:115 The urokinase plasminogen activator 81:115 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 1 16 theme urokinase 85:93 arg1 activator 107:115 The urokinase plasminogen activator 81:115 The urokinase plasminogen activator 81:115 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 4 17 theme antagonists 796:806 arg1 design 757:762 the design 753:762 the design of urokinase-urokinase receptor antagonists 753:806 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 4 18 with interaction 694:704 arg1 ligands 729:735 ligands 729:735 ligands 729:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 4 18 with interaction 694:704 arg1 variety 718:724 a wide variety 711:724 a wide variety of ligands 711:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 3 19 theme urokinase 571:579 arg1 fragment 581:588 the urokinase fragment 567:588 the urokinase fragment 567:588 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 0 20 theme activator 41:49 arg1 Structure 0:8 Structure 0:8 Structure of human urokinase plasminogen activator in complex with its receptor. 0:79 Structure of human urokinase plasminogen activator in complex with its receptor. 16456079 4 21 theme receptor 668:675 arg1 flexibility 639:649 the flexibility 635:649 the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists 635:806 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 0 22 theme plasminogen 29:39 arg1 activator 41:49 human urokinase plasminogen activator 13:49 human urokinase plasminogen activator 13:49 Structure of human urokinase plasminogen activator in complex with its receptor. 16456079 4 23 theme urokinase 658:666 arg1 receptor 668:675 the urokinase receptor 654:675 the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists 654:806 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 3 24 theme concave 513:519 arg1 shape 521:525 a concave shape 511:525 a concave shape 511:525 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 4 25 theme ligands 729:735 arg1 ligands 729:735 ligands 729:735 ligands 729:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 4 25 theme ligands 729:735 arg1 variety 718:724 a wide variety 711:724 a wide variety of ligands 711:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 4 26 with basis 743:747 arg1 ligands 729:735 ligands 729:735 ligands 729:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 4 26 with basis 743:747 arg1 variety 718:724 a wide variety 711:724 a wide variety of ligands 711:735 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 3 27 theme urokinase 487:495 arg1 receptor 497:504 urokinase receptor 487:504 urokinase receptor 487:504 The three domains of urokinase receptor form a concave shape with a central cone-shaped cavity where the urokinase fragment inserts. 16456079 1 28 theme pathological 223:234 arg1 metastasis 270:279 metastasis 270:279 metastasis 270:279 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 1 28 theme pathological 223:234 arg1 processes 236:244 pathological processes 223:244 pathological processes including tumor growth, metastasis, and inflammation 223:297 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 1 28 theme pathological 223:234 arg1 growth 262:267 tumor growth 256:267 tumor growth 256:267 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 1 28 theme pathological 223:234 arg1 inflammation 286:297 inflammation 286:297 inflammation 286:297 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 4 29 theme receptor 787:794 arg1 antagonists 796:806 urokinase-urokinase receptor antagonists 767:806 urokinase-urokinase receptor antagonists 767:806 The structure provides insight into the flexibility of the urokinase receptor that enables its interaction with a wide variety of ligands and a basis for the design of urokinase-urokinase receptor antagonists. 16456079 0 30 with complex 54:60 arg1 receptor 71:78 its receptor 67:78 its receptor 67:78 Structure of human urokinase plasminogen activator in complex with its receptor. 16456079 1 31 theme cellular 130:137 arg1 receptor 139:146 its cellular receptor 126:146 its cellular receptor with high affinity 126:165 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 16456079 0 32 theme human 13:17 arg1 activator 41:49 human urokinase plasminogen activator 13:49 human urokinase plasminogen activator 13:49 Structure of human urokinase plasminogen activator in complex with its receptor. 16456079 2 33 theme receptor 366:373 arg1 structure 322:330 the crystal structure 310:330 the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor 310:463 We report the crystal structure at 1.9 angstroms of the urokinase receptor complexed with the urokinase amino-terminal fragment and an antibody against the receptor. 16456079 1 34 theme tumor 256:260 arg1 growth 262:267 tumor growth 256:267 tumor growth 256:267 The urokinase plasminogen activator binds to its cellular receptor with high affinity and initiates signaling cascades that are implicated in pathological processes including tumor growth, metastasis, and inflammation. 3542989 5 0 theme cholinesterase 490:503 arg1 identical 514:522 identical 514:522 identical 514:522 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 5 0 theme cholinesterase 490:503 arg1 sequence 466:473 The sequence 462:473 The sequence of human serum cholinesterase 462:503 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 1 1 theme peptides 218:225 arg1 degradation 194:204 Edman degradation 188:204 Edman degradation of purified peptides 188:225 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 5 2 theme acetylcholinesterase 545:564 arg1 sequence 533:540 the sequence 529:540 the sequence of acetylcholinesterase from Torpedo californica 529:589 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 5 3 with identical 514:522 arg1 sequence 533:540 the sequence 529:540 the sequence of acetylcholinesterase from Torpedo californica 529:589 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 5 4 theme carboxyl-terminal 618:634 arg1 portion 636:642 the carboxyl-terminal portion 614:642 the carboxyl-terminal portion of bovine thyroglobulin 614:666 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 3 5 theme cholinesterase 354:367 arg1 subunits 342:349 The four subunits 333:349 The four subunits of cholinesterase 333:367 The four subunits of cholinesterase appear to be identical. 3542989 3 5 theme cholinesterase 354:367 arg1 identical 382:390 identical 382:390 identical 382:390 The four subunits of cholinesterase appear to be identical. 3542989 1 6 dep II 141:142 arg1 unspecific 145:154 unspecific 145:154 unspecific 145:154 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 2 7 attach attached 306:313 arg1 asparagines 320:330 9 asparagines 318:330 9 asparagines 318:330 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 2 7 attach attached 306:313 arg2 chains 299:304 nine carbohydrate chains 281:304 nine carbohydrate chains attached to 9 asparagines 281:330 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 4 8 from terminus 452:459 arg1 residue 429:435 the 198th residue 419:435 the 198th residue from the amino terminus 419:459 The active site serine is the 198th residue from the amino terminus. 3542989 4 8 from terminus 452:459 arg1 serine 409:414 The active site serine 393:414 The active site serine 393:414 The active site serine is the 198th residue from the amino terminus. 3542989 5 9 theme Torpedo 571:577 arg1 californica 579:589 Torpedo californica 571:589 Torpedo californica 571:589 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 2 10 theme amino 253:257 arg1 acids 259:263 574 amino acids 249:263 574 amino acids per subunit 249:275 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 2 11 theme carbohydrate 286:297 arg1 chains 299:304 nine carbohydrate chains 281:304 nine carbohydrate chains attached to 9 asparagines 281:330 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 1 12 theme complete 64:71 arg1 sequence 84:91 The complete amino acid sequence 60:91 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) 60:168 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 4 13 theme amino 446:450 arg1 terminus 452:459 the amino terminus 442:459 the amino terminus 442:459 The active site serine is the 198th residue from the amino terminus. 3542989 1 14 theme amino 73:77 arg1 sequence 84:91 The complete amino acid sequence 60:91 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) 60:168 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 0 15 theme amino 9:13 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of human serum cholinesterase 0:57 Complete amino acid sequence of human serum cholinesterase. 3542989 1 16 theme acid 79:82 arg1 sequence 84:91 The complete amino acid sequence 60:91 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) 60:168 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 0 17 theme Complete 0:7 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of human serum cholinesterase 0:57 Complete amino acid sequence of human serum cholinesterase. 3542989 2 18 contain contains 240:247 arg2 acids 259:263 574 amino acids 249:263 574 amino acids per subunit 249:275 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 2 18 contain contains 240:247 arg2 chains 299:304 nine carbohydrate chains 281:304 nine carbohydrate chains attached to 9 asparagines 281:330 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 2 18 contain contains 240:247 arg1 protein 232:238 The protein 228:238 The protein 228:238 The protein contains 574 amino acids per subunit and nine carbohydrate chains attached to 9 asparagines. 3542989 0 19 theme acid 15:18 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of human serum cholinesterase 0:57 Complete amino acid sequence of human serum cholinesterase. 3542989 1 20 theme human 96:100 arg1 cholinesterase 108:121 human serum cholinesterase 96:121 human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) 96:168 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 0 21 theme human 32:36 arg1 cholinesterase 44:57 human serum cholinesterase 32:57 human serum cholinesterase 32:57 Complete amino acid sequence of human serum cholinesterase. 3542989 5 22 with identical 599:607 arg1 portion 636:642 the carboxyl-terminal portion 614:642 the carboxyl-terminal portion of bovine thyroglobulin 614:666 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 4 23 theme 198th 423:427 arg1 residue 429:435 the 198th residue 419:435 the 198th residue from the amino terminus 419:459 The active site serine is the 198th residue from the amino terminus. 3542989 4 23 theme 198th 423:427 arg1 serine 409:414 The active site serine 393:414 The active site serine 393:414 The active site serine is the 198th residue from the amino terminus. 3542989 0 24 theme cholinesterase 44:57 arg1 sequence 20:27 Complete amino acid sequence 0:27 Complete amino acid sequence of human serum cholinesterase 0:57 Complete amino acid sequence of human serum cholinesterase. 3542989 1 25 theme serum 102:106 arg1 cholinesterase 108:121 human serum cholinesterase 96:121 human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) 96:168 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 0 26 theme serum 38:42 arg1 cholinesterase 44:57 human serum cholinesterase 32:57 human serum cholinesterase 32:57 Complete amino acid sequence of human serum cholinesterase. 3542989 5 27 theme bovine 647:652 arg1 thyroglobulin 654:666 bovine thyroglobulin 647:666 bovine thyroglobulin 647:666 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 1 28 theme cholinesterase 108:121 arg1 sequence 84:91 The complete amino acid sequence 60:91 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) 60:168 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 5 29 theme thyroglobulin 654:666 arg1 portion 636:642 the carboxyl-terminal portion 614:642 the carboxyl-terminal portion of bovine thyroglobulin 614:666 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 1 30 theme Edman 188:192 arg1 degradation 194:204 Edman degradation 188:204 Edman degradation of purified peptides 188:225 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 1 31 dep cholinesterase 108:121 arg1 EC 158:159 EC 3.1.1.8 158:167 EC 3.1.1.8 158:167 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 1 31 dep cholinesterase 108:121 arg1 II 141:142 choline esterase II 124:142 choline esterase II (unspecific) 124:155 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 4 32 theme site 404:407 arg1 residue 429:435 the 198th residue 419:435 the 198th residue from the amino terminus 419:459 The active site serine is the 198th residue from the amino terminus. 3542989 4 32 theme site 404:407 arg1 serine 409:414 The active site serine 393:414 The active site serine 393:414 The active site serine is the 198th residue from the amino terminus. 3542989 1 33 theme choline 124:130 arg1 EC 158:159 EC 3.1.1.8 158:167 EC 3.1.1.8 158:167 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 1 33 theme choline 124:130 arg1 II 141:142 choline esterase II 124:142 choline esterase II (unspecific) 124:155 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 4 34 theme active 397:402 arg1 residue 429:435 the 198th residue 419:435 the 198th residue from the amino terminus 419:459 The active site serine is the 198th residue from the amino terminus. 3542989 4 34 theme active 397:402 arg1 serine 409:414 The active site serine 393:414 The active site serine 393:414 The active site serine is the 198th residue from the amino terminus. 3542989 5 35 theme human 478:482 arg1 cholinesterase 490:503 human serum cholinesterase 478:503 human serum cholinesterase 478:503 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 1 36 theme esterase 132:139 arg1 EC 158:159 EC 3.1.1.8 158:167 EC 3.1.1.8 158:167 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 1 36 theme esterase 132:139 arg1 II 141:142 choline esterase II 124:142 choline esterase II (unspecific) 124:155 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 3542989 5 37 theme serum 484:488 arg1 cholinesterase 490:503 human serum cholinesterase 478:503 human serum cholinesterase 478:503 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 5 38 from californica 579:589 arg1 sequence 533:540 the sequence 529:540 the sequence of acetylcholinesterase from Torpedo californica 529:589 The sequence of human serum cholinesterase is 53.8% identical with the sequence of acetylcholinesterase from Torpedo californica and 28% identical with the carboxyl-terminal portion of bovine thyroglobulin. 3542989 1 39 theme purified 209:216 arg1 peptides 218:225 purified peptides 209:225 purified peptides 209:225 The complete amino acid sequence of human serum cholinesterase (choline esterase II (unspecific), EC 3.1.1.8) was determined by Edman degradation of purified peptides. 18077410 6 0 theme cleft 1171:1175 arg1 backside 1155:1162 the backside 1151:1162 the backside of the cleft 1151:1175 In contrast, Ab75 bound at the backside of the cleft to a region corresponding to thrombin exosite II, which is known to interact with allosteric effector molecules. 18077410 2 1 theme inhibitory 368:377 arg1 Ab75 400:403 Ab75 400:403 Ab75 400:403 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 1 theme inhibitory 368:377 arg1 Ab58 391:394 Ab58 391:394 Ab58 391:394 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 1 theme inhibitory 368:377 arg1 antibodies 379:388 The two inhibitory antibodies 360:388 The two inhibitory antibodies 360:388 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 9 2 theme catalytic 1737:1745 arg1 machinery 1747:1755 the catalytic machinery 1733:1755 the catalytic machinery of proteases 1733:1768 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 9 3 theme cleft 1709:1713 arg1 rim 1680:1682 the rim 1676:1682 the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops 1676:1807 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 7 4 theme competitive 1464:1474 arg1 inhibitor 1476:1484 a partial competitive inhibitor 1454:1484 a partial competitive inhibitor 1454:1484 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 7 4 theme competitive 1464:1474 arg1 Ab75 1446:1449 Ab75 1446:1449 Ab75 1446:1449 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 2 5 theme repertoire 619:628 arg1 diversity 593:601 the natural diversity 581:601 the natural diversity of the human Ig repertoire 581:628 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 7 6 theme active 1353:1358 arg1 inhibitors 1365:1374 active site inhibitors 1353:1374 active site inhibitors 1353:1374 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 2 7 theme chain 564:568 arg1 regions 526:532 the three complementarity determining regions 488:532 the three complementarity determining regions (H1, H2, and H3) of the heavy chain 488:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 7 theme chain 564:568 arg1 H2 539:540 H2 539:540 H2 539:540 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 7 theme chain 564:568 arg1 H3 547:548 H3 547:548 H3 547:548 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 7 theme chain 564:568 arg1 H1 535:536 H1 535:536 H1 535:536 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 8 theme Ig 616:617 arg1 repertoire 619:628 the human Ig repertoire 606:628 the human Ig repertoire 606:628 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 3 9 theme active 809:814 arg1 site 816:819 the active site 805:819 the active site 805:819 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 1 10 theme antibodies 273:282 arg1 structures 255:264 the structures 251:264 the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA) 251:357 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 7 11 theme binding 1333:1339 arg1 assays 1341:1346 binding assays 1333:1346 binding assays with active site inhibitors 1333:1374 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 2 12 theme heavy 558:562 arg1 chain 564:568 the heavy chain 554:568 the heavy chain 554:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 13 theme human 610:614 arg1 repertoire 619:628 the human Ig repertoire 606:628 the human Ig repertoire 606:628 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 7 14 theme site 1360:1363 arg1 inhibitors 1365:1374 active site inhibitors 1353:1374 active site inhibitors 1353:1374 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 8 15 theme structural 1509:1518 arg1 insight 1520:1526 structural insight 1509:1526 structural insight into antibody-mediated protease inhibition 1509:1569 These results provide structural insight into antibody-mediated protease inhibition. 18077410 8 16 theme protease 1551:1558 arg1 inhibition 1560:1569 antibody-mediated protease inhibition 1533:1569 antibody-mediated protease inhibition 1533:1569 These results provide structural insight into antibody-mediated protease inhibition. 18077410 2 17 theme determining 514:524 arg1 regions 526:532 the three complementarity determining regions 488:532 the three complementarity determining regions (H1, H2, and H3) of the heavy chain 488:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 17 theme determining 514:524 arg1 H2 539:540 H2 539:540 H2 539:540 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 17 theme determining 514:524 arg1 H3 547:548 H3 547:548 H3 547:548 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 17 theme determining 514:524 arg1 H1 535:536 H1 535:536 H1 535:536 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 18 from diversity 475:483 arg1 regions 526:532 the three complementarity determining regions 488:532 the three complementarity determining regions (H1, H2, and H3) of the heavy chain 488:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 18 from diversity 475:483 arg1 H2 539:540 H2 539:540 H2 539:540 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 18 from diversity 475:483 arg1 H3 547:548 H3 547:548 H3 547:548 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 18 from diversity 475:483 arg1 H1 535:536 H1 535:536 H1 535:536 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 7 19 with agreement 1293:1301 arg1 analysis 1323:1330 the structural analysis 1308:1330 the structural analysis 1308:1330 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 2 20 theme phage 438:442 arg1 library 452:458 a human Fab phage display library 426:458 a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain 426:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 7 21 with assays 1390:1395 arg1 inhibitors 1365:1374 active site inhibitors 1353:1374 active site inhibitors 1353:1374 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 1 22 theme concave 183:189 arg1 clefts 216:221 the concave shaped substrate-binding clefts 179:221 the concave shaped substrate-binding clefts of proteases 179:234 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 2 23 theme display 444:450 arg1 library 452:458 a human Fab phage display library 426:458 a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain 426:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 24 theme natural 585:591 arg1 diversity 593:601 the natural diversity 581:601 the natural diversity of the human Ig repertoire 581:628 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 6 25 theme allosteric 1259:1268 arg1 molecules 1279:1287 allosteric effector molecules 1259:1287 allosteric effector molecules 1259:1287 In contrast, Ab75 bound at the backside of the cleft to a region corresponding to thrombin exosite II, which is known to interact with allosteric effector molecules. 18077410 1 26 theme shaped 191:196 arg1 clefts 216:221 the concave shaped substrate-binding clefts 179:221 the concave shaped substrate-binding clefts of proteases 179:234 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 0 27 theme Structural 0:9 arg1 insight 11:17 Structural insight 0:17 Structural insight into distinct mechanisms of protease inhibition by antibodies 0:79 Structural insight into distinct mechanisms of protease inhibition by antibodies. 18077410 5 28 theme substrate 1083:1091 arg1 sites 1105:1109 important substrate interaction sites 1073:1109 important substrate interaction sites 1073:1109 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 7 29 theme enzymatic 1380:1388 arg1 assays 1390:1395 enzymatic assays 1380:1395 enzymatic assays 1380:1395 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 1 30 with complex 287:293 arg1 HGFA 353:356 HGFA 353:356 HGFA 353:356 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 30 with complex 287:293 arg1 activator 342:350 the trypsin-like hepatocyte growth-factor activator 300:350 the trypsin-like hepatocyte growth-factor activator (HGFA) 300:357 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 9 31 theme protruding 1656:1665 arg1 loops 1667:1671 protruding loops 1656:1671 protruding loops 1656:1671 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 1 32 theme substrate-binding 198:214 arg1 clefts 216:221 the concave shaped substrate-binding clefts 179:221 the concave shaped substrate-binding clefts of proteases 179:234 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 0 33 theme distinct 24:31 arg1 mechanisms 33:42 distinct mechanisms 24:42 distinct mechanisms of protease inhibition by antibodies 24:79 Structural insight into distinct mechanisms of protease inhibition by antibodies. 18077410 3 34 dep structures 659:668 arg1 HGFA 683:686 HGFA 683:686 HGFA (3.5-A resolution) 683:705 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 3 34 dep structures 659:668 arg1 resolution 695:704 3.5-A resolution 689:704 3.5-A resolution 689:704 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 3 34 dep structures 659:668 arg1 complexes 745:753 the Fab75:HGFA (2.2-A resolution) complexes 711:753 the Fab75:HGFA (2.2-A resolution) complexes 711:753 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 2 35 dep regions 526:532 arg1 regions 526:532 the three complementarity determining regions 488:532 the three complementarity determining regions (H1, H2, and H3) of the heavy chain 488:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 35 dep regions 526:532 arg1 H2 539:540 H2 539:540 H2 539:540 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 35 dep regions 526:532 arg1 H3 547:548 H3 547:548 H3 547:548 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 35 dep regions 526:532 arg1 H1 535:536 H1 535:536 H1 535:536 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 8 36 theme antibody-mediated 1533:1549 arg1 inhibition 1560:1569 antibody-mediated protease inhibition 1533:1569 antibody-mediated protease inhibition 1533:1569 These results provide structural insight into antibody-mediated protease inhibition. 18077410 2 37 theme human 428:432 arg1 library 452:458 a human Fab phage display library 426:458 a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain 426:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 3 38 theme 3.5-A 689:693 arg1 HGFA 683:686 HGFA 683:686 HGFA (3.5-A resolution) 683:705 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 3 38 theme 3.5-A 689:693 arg1 resolution 695:704 3.5-A resolution 689:704 3.5-A resolution 689:704 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 6 39 theme effector 1270:1277 arg1 molecules 1279:1287 allosteric effector molecules 1259:1287 allosteric effector molecules 1259:1287 In contrast, Ab75 bound at the backside of the cleft to a region corresponding to thrombin exosite II, which is known to interact with allosteric effector molecules. 18077410 5 40 theme H1 1034:1035 arg1 loops 1044:1048 its H1 and H2 loops 1030:1048 its H1 and H2 loops in the cleft 1030:1061 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 3 41 theme substrate 860:868 arg1 hydrolysis 870:879 substrate hydrolysis 860:879 substrate hydrolysis 860:879 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 4 42 theme protruding 937:946 arg1 element 948:954 the same protruding element 928:954 the same protruding element (99-loop) 928:964 In both cases, the antibodies interacted with the same protruding element (99-loop), which forms part of the substrate-binding cleft. 18077410 4 42 theme protruding 937:946 arg1 99-loop 957:963 99-loop 957:963 99-loop 957:963 In both cases, the antibodies interacted with the same protruding element (99-loop), which forms part of the substrate-binding cleft. 18077410 1 43 theme trypsin-like 304:315 arg1 HGFA 353:356 HGFA 353:356 HGFA 353:356 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 43 theme trypsin-like 304:315 arg1 activator 342:350 the trypsin-like hepatocyte growth-factor activator 300:350 the trypsin-like hepatocyte growth-factor activator (HGFA) 300:357 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 6 44 theme thrombin 1206:1213 arg1 exosite 1215:1221 thrombin exosite II 1206:1224 thrombin exosite II 1206:1224 In contrast, Ab75 bound at the backside of the cleft to a region corresponding to thrombin exosite II, which is known to interact with allosteric effector molecules. 18077410 7 45 theme structural 1312:1321 arg1 analysis 1323:1330 the structural analysis 1308:1330 the structural analysis 1308:1330 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 1 46 theme proteases 226:234 arg1 clefts 216:221 the concave shaped substrate-binding clefts 179:221 the concave shaped substrate-binding clefts of proteases 179:234 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 2 47 theme synthetic 465:473 arg1 diversity 475:483 synthetic diversity 465:483 synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain 465:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 1 48 theme hepatocyte 317:326 arg1 HGFA 353:356 HGFA 353:356 HGFA 353:356 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 48 theme hepatocyte 317:326 arg1 activator 342:350 the trypsin-like hepatocyte growth-factor activator 300:350 the trypsin-like hepatocyte growth-factor activator (HGFA) 300:357 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 4 49 theme same 932:935 arg1 element 948:954 the same protruding element 928:954 the same protruding element (99-loop) 928:964 In both cases, the antibodies interacted with the same protruding element (99-loop), which forms part of the substrate-binding cleft. 18077410 4 49 theme same 932:935 arg1 99-loop 957:963 99-loop 957:963 99-loop 957:963 In both cases, the antibodies interacted with the same protruding element (99-loop), which forms part of the substrate-binding cleft. 18077410 2 50 theme Fab 434:436 arg1 library 452:458 a human Fab phage display library 426:458 a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain 426:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 5 51 theme important 1073:1081 arg1 sites 1105:1109 important substrate interaction sites 1073:1109 important substrate interaction sites 1073:1109 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 0 52 theme inhibition 56:65 arg1 mechanisms 33:42 distinct mechanisms 24:42 distinct mechanisms of protease inhibition by antibodies 24:79 Structural insight into distinct mechanisms of protease inhibition by antibodies. 18077410 1 53 theme growth-factor 328:340 arg1 HGFA 353:356 HGFA 353:356 HGFA 353:356 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 53 theme growth-factor 328:340 arg1 activator 342:350 the trypsin-like hepatocyte growth-factor activator 300:350 the trypsin-like hepatocyte growth-factor activator (HGFA) 300:357 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 5 54 theme interaction 1093:1103 arg1 sites 1105:1109 important substrate interaction sites 1073:1109 important substrate interaction sites 1073:1109 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 3 55 theme Fab58 677:681 arg1 studies 643:649 Biochemical studies 631:649 Biochemical studies 631:649 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 3 55 theme Fab58 677:681 arg1 structures 659:668 the structures 655:668 the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes 655:753 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 0 56 theme protease 47:54 arg1 inhibition 56:65 protease inhibition 47:65 protease inhibition by antibodies 47:79 Structural insight into distinct mechanisms of protease inhibition by antibodies. 18077410 5 57 dep inserted 1021:1028 arg1 S2 1119:1120 S2 1119:1120 S2 1119:1120 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 5 57 dep inserted 1021:1028 arg1 S3 1112:1113 S3 1112:1113 S3 1112:1113 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 1 58 from structures 255:264 arg1 complex 287:293 complex 287:293 complex with the trypsin-like hepatocyte growth-factor activator (HGFA) 287:357 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 7 59 with assays 1341:1346 arg1 inhibitors 1365:1374 active site inhibitors 1353:1374 active site inhibitors 1353:1374 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 9 60 theme substrate-binding 1691:1707 arg1 cleft 1709:1713 the substrate-binding cleft 1687:1713 the substrate-binding cleft 1687:1713 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 1 61 theme flat 122:125 arg1 antibodies 154:163 antibodies 154:163 antibodies 154:163 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 61 theme flat 122:125 arg1 sites 145:149 the relatively flat antigen-combining sites 107:149 the relatively flat antigen-combining sites of antibodies 107:163 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 4 62 theme cleft 1009:1013 arg1 part 979:982 part 979:982 part of the substrate-binding cleft 979:1013 In both cases, the antibodies interacted with the same protruding element (99-loop), which forms part of the substrate-binding cleft. 18077410 3 63 theme Fab75 715:719 arg1 resolution 733:742 2.2-A resolution 727:742 2.2-A resolution 727:742 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 3 63 theme Fab75 715:719 arg1 HGFA 721:724 the Fab75:HGFA 711:724 the Fab75:HGFA (2.2-A resolution) complexes 711:753 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 9 64 theme long 1788:1791 arg1 loops 1803:1807 long insertion loops 1788:1807 long insertion loops 1788:1807 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 1 65 theme antigen-combining 127:143 arg1 antibodies 154:163 antibodies 154:163 antibodies 154:163 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 65 theme antigen-combining 127:143 arg1 sites 145:149 the relatively flat antigen-combining sites 107:149 the relatively flat antigen-combining sites of antibodies 107:163 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 4 66 theme substrate-binding 991:1007 arg1 cleft 1009:1013 the substrate-binding cleft 987:1013 the substrate-binding cleft 987:1013 In both cases, the antibodies interacted with the same protruding element (99-loop), which forms part of the substrate-binding cleft. 18077410 9 67 theme canonical 1597:1605 arg1 inhibitors 1607:1616 canonical inhibitors 1597:1616 canonical inhibitors 1597:1616 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 7 68 theme partial 1456:1462 arg1 inhibitor 1476:1484 a partial competitive inhibitor 1454:1484 a partial competitive inhibitor 1454:1484 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 7 68 theme partial 1456:1462 arg1 Ab75 1446:1449 Ab75 1446:1449 Ab75 1446:1449 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 2 69 theme complementarity 498:512 arg1 regions 526:532 the three complementarity determining regions 488:532 the three complementarity determining regions (H1, H2, and H3) of the heavy chain 488:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 69 theme complementarity 498:512 arg1 H2 539:540 H2 539:540 H2 539:540 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 69 theme complementarity 498:512 arg1 H3 547:548 H3 547:548 H3 547:548 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 2 69 theme complementarity 498:512 arg1 H1 535:536 H1 535:536 H1 535:536 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 3 70 theme Biochemical 631:641 arg1 studies 643:649 Biochemical studies 631:649 Biochemical studies 631:649 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 2 71 with library 452:458 arg1 diversity 475:483 synthetic diversity 465:483 synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain 465:568 The two inhibitory antibodies, Ab58 and Ab75, were generated from a human Fab phage display library with synthetic diversity in the three complementarity determining regions (H1, H2, and H3) of the heavy chain, mimicking the natural diversity of the human Ig repertoire. 18077410 5 72 from loops 1044:1048 arg1 cleft 1057:1061 the cleft 1053:1061 the cleft 1053:1061 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 3 73 theme substrate 785:793 arg1 access 795:800 substrate access 785:800 substrate access to the active site 785:819 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 7 74 theme competitive 1419:1429 arg1 Ab58 1409:1412 Ab58 1409:1412 Ab58 1409:1412 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 7 74 theme competitive 1419:1429 arg1 inhibitor 1431:1439 a competitive inhibitor 1417:1439 a competitive inhibitor 1417:1439 In agreement with the structural analysis, binding assays with active site inhibitors and enzymatic assays showed that Ab58 is a competitive inhibitor, and Ab75 is a partial competitive inhibitor. 18077410 5 75 theme H2 1041:1042 arg1 loops 1044:1048 its H1 and H2 loops 1030:1048 its H1 and H2 loops in the cleft 1030:1061 Ab58 inserted its H1 and H2 loops in the cleft to occupy important substrate interaction sites (S3 and S2). 18077410 3 76 theme 2.2-A 727:731 arg1 resolution 733:742 2.2-A resolution 727:742 2.2-A resolution 727:742 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 3 76 theme 2.2-A 727:731 arg1 HGFA 721:724 the Fab75:HGFA 711:724 the Fab75:HGFA (2.2-A resolution) complexes 711:753 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 18077410 9 77 theme proteases 1760:1768 arg1 machinery 1747:1755 the catalytic machinery 1733:1755 the catalytic machinery of proteases 1733:1768 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 1 78 theme antibodies 154:163 arg1 antibodies 154:163 antibodies 154:163 antibodies 154:163 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 1 78 theme antibodies 154:163 arg1 sites 145:149 the relatively flat antigen-combining sites 107:149 the relatively flat antigen-combining sites of antibodies 107:163 To better understand how the relatively flat antigen-combining sites of antibodies interact with the concave shaped substrate-binding clefts of proteases, we determined the structures of two antibodies in complex with the trypsin-like hepatocyte growth-factor activator (HGFA). 18077410 9 79 theme insertion 1793:1801 arg1 loops 1803:1807 long insertion loops 1788:1807 long insertion loops 1788:1807 They suggest that unlike canonical inhibitors, antibodies may preferentially target protruding loops at the rim of the substrate-binding cleft to interfere with the catalytic machinery of proteases without requiring long insertion loops. 18077410 3 80 theme HGFA 721:724 arg1 complexes 745:753 the Fab75:HGFA (2.2-A resolution) complexes 711:753 the Fab75:HGFA (2.2-A resolution) complexes 711:753 Biochemical studies and the structures of the Fab58:HGFA (3.5-A resolution) and the Fab75:HGFA (2.2-A resolution) complexes revealed that Ab58 obstructed substrate access to the active site, whereas Ab75 allosterically inhibited substrate hydrolysis. 24036510 3 0 contain containing 444:453 arg2 site 469:472 the catalytic site 455:472 the catalytic site 455:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 0 contain containing 444:453 arg1 domain 488:493 a β-sandwich domain 475:493 a β-sandwich domain 475:493 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 0 contain containing 444:453 arg1 domain 437:442 a TIM barrel domain 424:442 a TIM barrel domain containing the catalytic site 424:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 0 contain containing 444:453 arg1 domain 518:523 a fibronectin-like domain 499:523 a fibronectin-like domain 499:523 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 6 1 theme IDUA 951:954 arg1 structures 956:965 these IDUA structures 945:965 these IDUA structures 945:965 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 0 2 theme α-L-iduronidase 71:85 arg1 action 61:66 action 61:66 action 61:66 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase. 24036510 0 2 theme α-L-iduronidase 71:85 arg1 structure 47:55 structure 47:55 structure 47:55 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase. 24036510 1 3 from mutations 136:144 arg1 gene 153:156 the gene 149:156 the gene encoding α-L-iduronidase (IDUA) 149:188 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 4 4 theme B 623:623 arg1 conformation 625:636 a (2,5)B conformation 616:636 a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182 616:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 1 5 theme genetic 218:224 arg1 disorders 226:234 approximately 70 genetic disorders 201:234 approximately 70 genetic disorders collectively known as the lysosomal storage diseases 201:287 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 3 6 theme β-sandwich 477:486 arg1 domain 488:493 a β-sandwich domain 475:493 a β-sandwich domain 475:493 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 6 theme β-sandwich 477:486 arg1 domain 437:442 a TIM barrel domain 424:442 a TIM barrel domain containing the catalytic site 424:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 1 7 theme disorders 226:234 arg1 disorders 226:234 approximately 70 genetic disorders 201:234 approximately 70 genetic disorders collectively known as the lysosomal storage diseases 201:287 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 1 7 theme disorders 226:234 arg1 one 194:196 one 194:196 one 194:196 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 1 8 theme Mucopolysaccharidosis 88:108 arg1 type 110:113 Mucopolysaccharidosis type I 88:115 Mucopolysaccharidosis type I (MPS I) 88:123 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 1 8 theme Mucopolysaccharidosis 88:108 arg1 MPS 118:120 MPS 118:120 MPS 118:120 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 5 9 theme enzymatic 916:924 arg1 activity 926:933 enzymatic activity 916:933 enzymatic activity 916:933 Unexpectedly, the N-glycan attached to Asn372 interacts with iduronate analogs in the active site and is required for enzymatic activity. 24036510 4 10 theme retaining 691:699 arg1 reaction 721:728 a retaining double displacement reaction 689:728 a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182 689:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 4 11 theme glycosyl-enzyme 645:659 arg1 intermediate 661:672 the glycosyl-enzyme intermediate 641:672 the glycosyl-enzyme intermediate 641:672 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 0 12 from structure 47:55 arg1 Insights 0:7 Insights 0:7 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase 0:85 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase. 24036510 2 13 theme human 348:352 arg1 IDUA 354:357 human IDUA 348:357 human IDUA produced in an Arabidopsis thaliana cgl mutant 348:404 To gain insight into the basis for MPS I, we crystallized human IDUA produced in an Arabidopsis thaliana cgl mutant. 24036510 2 14 dep Arabidopsis 374:384 arg1 thaliana 386:393 thaliana 386:393 thaliana 386:393 To gain insight into the basis for MPS I, we crystallized human IDUA produced in an Arabidopsis thaliana cgl mutant. 24036510 4 15 theme displacement 708:719 arg1 reaction 721:728 a retaining double displacement reaction 689:728 a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182 689:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 3 16 theme fibronectin-like 501:516 arg1 domain 437:442 a TIM barrel domain 424:442 a TIM barrel domain containing the catalytic site 424:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 16 theme fibronectin-like 501:516 arg1 domain 518:523 a fibronectin-like domain 499:523 a fibronectin-like domain 499:523 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 2 17 theme cgl 395:397 arg1 mutant 399:404 an Arabidopsis thaliana cgl mutant 371:404 an Arabidopsis thaliana cgl mutant 371:404 To gain insight into the basis for MPS I, we crystallized human IDUA produced in an Arabidopsis thaliana cgl mutant. 24036510 5 18 from analogs 869:875 arg1 site 891:894 the active site 880:894 the active site 880:894 Unexpectedly, the N-glycan attached to Asn372 interacts with iduronate analogs in the active site and is required for enzymatic activity. 24036510 5 19 attach attached 825:832 arg2 N-glycan 816:823 the N-glycan 812:823 the N-glycan attached to Asn372 812:842 Unexpectedly, the N-glycan attached to Asn372 interacts with iduronate analogs in the active site and is required for enzymatic activity. 24036510 5 19 attach attached 825:832 arg1 Asn372 837:842 Asn372 837:842 Asn372 837:842 Unexpectedly, the N-glycan attached to Asn372 interacts with iduronate analogs in the active site and is required for enzymatic activity. 24036510 6 20 theme mutation 1022:1029 arg1 structures 956:965 these IDUA structures 945:965 these IDUA structures 945:965 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 6 20 theme mutation 1022:1029 arg1 analysis 983:990 biochemical analysis 971:990 biochemical analysis of the disease-relevant P533R mutation 971:1029 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 5 21 theme active 884:889 arg1 site 891:894 the active site 880:894 the active site 880:894 Unexpectedly, the N-glycan attached to Asn372 interacts with iduronate analogs in the active site and is required for enzymatic activity. 24036510 6 22 theme clinical 1096:1103 arg1 phenotypes 1105:1114 clinical phenotypes 1096:1114 clinical phenotypes 1096:1114 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 6 23 theme P533R 1016:1020 arg1 mutation 1022:1029 the disease-relevant P533R mutation 995:1029 the disease-relevant P533R mutation 995:1029 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 2 24 theme Arabidopsis 374:384 arg1 mutant 399:404 an Arabidopsis thaliana cgl mutant 371:404 an Arabidopsis thaliana cgl mutant 371:404 To gain insight into the basis for MPS I, we crystallized human IDUA produced in an Arabidopsis thaliana cgl mutant. 24036510 5 25 theme iduronate 859:867 arg1 analogs 869:875 iduronate analogs 859:875 iduronate analogs in the active site 859:894 Unexpectedly, the N-glycan attached to Asn372 interacts with iduronate analogs in the active site and is required for enzymatic activity. 24036510 6 26 theme disease-relevant 999:1014 arg1 mutation 1022:1029 the disease-relevant P533R mutation 995:1029 the disease-relevant P533R mutation 995:1029 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 1 27 theme lysosomal 262:270 arg1 diseases 280:287 the lysosomal storage diseases 258:287 the lysosomal storage diseases 258:287 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 1 28 theme storage 272:278 arg1 diseases 280:287 the lysosomal storage diseases 258:287 the lysosomal storage diseases 258:287 Mucopolysaccharidosis type I (MPS I), caused by mutations in the gene encoding α-L-iduronidase (IDUA), is one of approximately 70 genetic disorders collectively known as the lysosomal storage diseases. 24036510 0 29 dep structure 47:55 arg1 the 43:45 the 43:45 the 43:45 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase. 24036510 6 30 from mutations 1075:1083 arg1 IDUA 1088:1091 IDUA 1088:1091 IDUA to clinical phenotypes 1088:1114 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 4 31 theme Michaelis 587:595 arg1 complex 597:603 the Michaelis complex 583:603 the Michaelis complex 583:603 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 0 32 from action 61:66 arg1 Insights 0:7 Insights 0:7 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase 0:85 Insights into mucopolysaccharidosis I from the structure and action of α-L-iduronidase. 24036510 4 33 theme acid/base 780:788 arg1 Glu182 790:795 the general acid/base Glu182 768:795 the general acid/base Glu182 768:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 3 34 theme barrel 430:435 arg1 domain 488:493 a β-sandwich domain 475:493 a β-sandwich domain 475:493 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 34 theme barrel 430:435 arg1 domain 437:442 a TIM barrel domain 424:442 a TIM barrel domain containing the catalytic site 424:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 34 theme barrel 430:435 arg1 domain 518:523 a fibronectin-like domain 499:523 a fibronectin-like domain 499:523 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 4 35 theme general 772:778 arg1 Glu182 790:795 the general acid/base Glu182 768:795 the general acid/base Glu182 768:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 6 36 from IDUA 1088:1091 arg1 effects 1064:1070 the effects 1060:1070 the effects of mutations in IDUA to clinical phenotypes 1060:1114 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 6 37 theme mutations 1075:1083 arg1 effects 1064:1070 the effects 1060:1070 the effects of mutations in IDUA to clinical phenotypes 1060:1114 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 4 38 theme double 701:706 arg1 reaction 721:728 a retaining double displacement reaction 689:728 a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182 689:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 3 39 theme TIM 426:428 arg1 domain 488:493 a β-sandwich domain 475:493 a β-sandwich domain 475:493 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 39 theme TIM 426:428 arg1 domain 437:442 a TIM barrel domain 424:442 a TIM barrel domain containing the catalytic site 424:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 3 39 theme TIM 426:428 arg1 domain 518:523 a fibronectin-like domain 499:523 a fibronectin-like domain 499:523 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 6 40 theme biochemical 971:981 arg1 analysis 983:990 biochemical analysis 971:990 biochemical analysis of the disease-relevant P533R mutation 971:1029 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 3 41 theme catalytic 459:467 arg1 site 469:472 the catalytic site 455:472 the catalytic site 455:472 IDUA consists of a TIM barrel domain containing the catalytic site, a β-sandwich domain and a fibronectin-like domain. 24036510 4 42 theme IDUA 540:543 arg1 Structures 526:535 Structures 526:535 Structures of IDUA bound to iduronate analogs 526:570 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 4 43 theme nucleophilic 744:755 arg1 Glu299 757:762 the nucleophilic Glu299 740:762 the nucleophilic Glu299 740:762 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 6 44 from effects 1064:1070 arg1 IDUA 1088:1091 IDUA 1088:1091 IDUA to clinical phenotypes 1088:1114 Finally, these IDUA structures and biochemical analysis of the disease-relevant P533R mutation have enabled us to correlate the effects of mutations in IDUA to clinical phenotypes. 24036510 4 45 theme 2,5 619:621 arg1 conformation 625:636 a (2,5)B conformation 616:636 a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182 616:795 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 4 46 from conformation 625:636 arg1 intermediate 661:672 the glycosyl-enzyme intermediate 641:672 the glycosyl-enzyme intermediate 641:672 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 24036510 4 47 theme iduronate 554:562 arg1 analogs 564:570 iduronate analogs 554:570 iduronate analogs 554:570 Structures of IDUA bound to iduronate analogs illustrate the Michaelis complex and reveal a (2,5)B conformation in the glycosyl-enzyme intermediate, which suggest a retaining double displacement reaction involving the nucleophilic Glu299 and the general acid/base Glu182. 29899144 0 0 theme lipoprotein 78:88 arg1 lipase 90:95 lipoprotein lipase 78:95 lipoprotein lipase 78:95 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine regulates lipoprotein lipase. 29899144 1 1 theme lipoprotein 172:182 arg1 protein 221:227 a membrane protein 210:227 a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen 210:335 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 1 1 theme lipoprotein 172:182 arg1 LPL 192:194 LPL 192:194 LPL 192:194 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 1 1 theme lipoprotein 172:182 arg1 lipase 184:189 lipoprotein lipase 172:189 lipoprotein lipase (LPL) 172:195 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 5 2 theme conserved 756:764 arg1 tyrosine 766:773 a conserved tyrosine 754:773 a conserved tyrosine in the middle of the IDR 754:798 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 5 3 from tyrosine 766:773 arg1 middle 782:787 the middle 778:787 the middle of the IDR 778:798 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 9 4 theme autoimmune 1553:1562 arg1 syndrome 1564:1571 the GPIHBP1 autoimmune syndrome 1541:1571 the GPIHBP1 autoimmune syndrome 1541:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 9 5 from factor 1477:1482 arg1 pathogenicity 1491:1503 the pathogenicity 1487:1503 the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome 1487:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 6 6 theme association 1131:1141 arg1 kon 1158:1160 kon 1158:1160 kon 1158:1160 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 6 6 theme association 1131:1141 arg1 constant 1148:1155 the association rate constant 1127:1155 the association rate constant (kon) for LPL binding 1127:1177 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 8 7 theme LPL 1348:1350 arg1 accumulation 1332:1343 the accumulation 1328:1343 the accumulation of LPL in close proximity to capillaries 1328:1384 In wild-type mice, we expect that the accumulation of LPL in close proximity to capillaries would increase interactions with GPIHBP1. 29899144 6 8 theme electrostatic 1092:1104 arg1 steering 1106:1113 electrostatic steering 1092:1113 electrostatic steering 1092:1113 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 9 9 theme key 1473:1475 arg1 factor 1477:1482 a key factor 1471:1482 a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome 1471:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 9 9 theme key 1473:1475 arg1 IDR 1460:1462 GPIHBP1's IDR 1450:1462 GPIHBP1's IDR 1450:1462 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 3 10 theme rich 554:557 arg1 IDR 549:551 IDR 549:551 IDR 549:551 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 10 theme rich 554:557 arg1 region 541:546 an intrinsically disordered region 513:546 an intrinsically disordered region (IDR) rich in acidic residues 513:576 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 10 theme rich 554:557 arg1 GPIHBP1 504:510 GPIHBP1 504:510 GPIHBP1 504:510 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 10 11 dep traverses 1667:1675 arg1 contributing 1769:1780 contributing 1769:1780 simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity 1754:1840 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 10 11 dep traverses 1667:1675 arg1 facilitating 1691:1702 facilitating 1691:1702 facilitating capture of LPL by capillary endothelial cells 1691:1748 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 3 12 theme GPIHBP1 504:510 arg1 domain 494:499 The N-terminal domain 479:499 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, 479:577 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 12 theme GPIHBP1 504:510 arg1 important 582:590 important 582:590 important 582:590 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 12 theme GPIHBP1 504:510 arg1 GPIHBP1 504:510 GPIHBP1 504:510 GPIHBP1 504:510 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 13 theme N-terminal 483:492 arg1 domain 494:499 The N-terminal domain 479:499 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, 479:577 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 13 theme N-terminal 483:492 arg1 important 582:590 important 582:590 important 582:590 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 13 theme N-terminal 483:492 arg1 GPIHBP1 504:510 GPIHBP1 504:510 GPIHBP1 504:510 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 5 14 theme GPIHBP1-LPL 897:907 arg1 interactions 909:920 GPIHBP1-LPL interactions 897:920 GPIHBP1-LPL interactions 897:920 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 2 15 dep GPIHBP1 356:362 arg1 absence 345:351 absence 345:351 absence 345:351 In the absence of GPIHBP1, LPL remains mislocalized within the subendothelial spaces, causing severe hypertriglyceridemia (chylomicronemia). 29899144 2 15 dep GPIHBP1 356:362 arg1 the 341:343 the 341:343 the 341:343 In the absence of GPIHBP1, LPL remains mislocalized within the subendothelial spaces, causing severe hypertriglyceridemia (chylomicronemia). 29899144 6 16 theme encounter 1078:1086 arg1 probability 1049:1059 the probability 1045:1059 the probability of a GPIHBP1-LPL encounter 1045:1086 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 5 17 theme ANGPTL4-catalyzed 972:988 arg1 unfolding 990:998 ANGPTL4-catalyzed unfolding 972:998 ANGPTL4-catalyzed unfolding 972:998 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 6 18 theme GPIHBP1-LPL 1066:1076 arg1 encounter 1078:1086 a GPIHBP1-LPL encounter 1064:1086 a GPIHBP1-LPL encounter 1064:1086 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 0 19 theme disordered 2:11 arg1 domain 20:25 A disordered acidic domain 0:25 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine 0:66 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine regulates lipoprotein lipase. 29899144 10 20 theme capillary 1722:1730 arg1 cells 1744:1748 capillary endothelial cells 1722:1748 capillary endothelial cells 1722:1748 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 10 21 theme endothelial 1732:1742 arg1 cells 1744:1748 capillary endothelial cells 1722:1748 capillary endothelial cells 1722:1748 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 2 22 theme severe 432:437 arg1 chylomicronemia 461:475 chylomicronemia 461:475 chylomicronemia 461:475 In the absence of GPIHBP1, LPL remains mislocalized within the subendothelial spaces, causing severe hypertriglyceridemia (chylomicronemia). 29899144 2 22 theme severe 432:437 arg1 hypertriglyceridemia 439:458 severe hypertriglyceridemia 432:458 severe hypertriglyceridemia (chylomicronemia) 432:476 In the absence of GPIHBP1, LPL remains mislocalized within the subendothelial spaces, causing severe hypertriglyceridemia (chylomicronemia). 29899144 10 23 theme charged 1644:1650 arg1 IDR 1652:1654 the negatively charged IDR 1629:1654 the negatively charged IDR of GPIHBP1 1629:1665 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 3 24 theme acidic 562:567 arg1 residues 569:576 acidic residues 562:576 acidic residues 562:576 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 25 from residues 569:576 arg1 rich 554:557 rich 554:557 rich 554:557 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 9 26 from pathogenicity 1491:1503 arg1 patients 1527:1534 patients 1527:1534 patients with the GPIHBP1 autoimmune syndrome 1527:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 5 27 theme IDR 796:798 arg1 middle 782:787 the middle 778:787 the middle of the IDR 778:798 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 1 28 theme subendothelial 276:289 arg1 spaces 291:296 the subendothelial spaces 272:296 the subendothelial spaces 272:296 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 5 29 theme GPIHBP1 941:947 arg1 affinity 885:892 the affinity 881:892 the affinity of GPIHBP1-LPL interactions 881:920 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 5 29 theme GPIHBP1 941:947 arg1 ability 930:936 the ability 926:936 the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding 926:998 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 10 30 theme LPL 1815:1817 arg1 structure 1819:1827 LPL structure 1815:1827 LPL structure 1815:1827 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 0 31 theme acidic 13:18 arg1 domain 20:25 A disordered acidic domain 0:25 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine 0:66 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine regulates lipoprotein lipase. 29899144 3 32 theme disordered 530:539 arg1 IDR 549:551 IDR 549:551 IDR 549:551 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 32 theme disordered 530:539 arg1 region 541:546 an intrinsically disordered region 513:546 an intrinsically disordered region (IDR) rich in acidic residues 513:576 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 3 32 theme disordered 530:539 arg1 GPIHBP1 504:510 GPIHBP1 504:510 GPIHBP1 504:510 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 1 33 theme intravascular 102:114 arg1 processing 116:125 The intravascular processing 98:125 The intravascular processing of triglyceride-rich lipoproteins 98:159 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 2 34 theme subendothelial 401:414 arg1 spaces 416:421 the subendothelial spaces 397:421 the subendothelial spaces 397:421 In the absence of GPIHBP1, LPL remains mislocalized within the subendothelial spaces, causing severe hypertriglyceridemia (chylomicronemia). 29899144 5 35 theme interactions 909:920 arg1 affinity 885:892 the affinity 881:892 the affinity of GPIHBP1-LPL interactions 881:920 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 5 35 theme interactions 909:920 arg1 ability 930:936 the ability 926:936 the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding 926:998 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 8 36 from accumulation 1332:1343 arg1 proximity 1361:1369 close proximity 1355:1369 close proximity to capillaries 1355:1384 In wild-type mice, we expect that the accumulation of LPL in close proximity to capillaries would increase interactions with GPIHBP1. 29899144 3 37 theme spontaneous 639:649 arg1 unfolding 673:681 spontaneous and ANGPTL4-catalyzed unfolding 639:681 spontaneous and ANGPTL4-catalyzed unfolding 639:681 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 9 38 theme GPIHBP1 1545:1551 arg1 syndrome 1564:1571 the GPIHBP1 autoimmune syndrome 1541:1571 the GPIHBP1 autoimmune syndrome 1541:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 10 39 theme biophysical 1592:1602 arg1 studies 1604:1610 biophysical studies 1592:1610 biophysical studies 1592:1610 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 5 40 mod modified 823:830 arg1 tyrosine 766:773 a conserved tyrosine 754:773 a conserved tyrosine in the middle of the IDR 754:798 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 5 40 mod modified 823:830 arg3 O-sulfation 835:845 O-sulfation 835:845 O-sulfation 835:845 First, a conserved tyrosine in the middle of the IDR is posttranslationally modified by O-sulfation; this modification increases both the affinity of GPIHBP1-LPL interactions and the ability of GPIHBP1 to protect LPL against ANGPTL4-catalyzed unfolding. 29899144 3 41 theme catalytic 614:622 arg1 domain 624:629 LPL's catalytic domain 608:629 LPL's catalytic domain 608:629 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 6 42 theme acidic 1013:1018 arg1 IDR 1020:1022 the acidic IDR 1009:1022 the acidic IDR of GPIHBP1 1009:1033 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 1 43 theme membrane 212:219 arg1 protein 221:227 a membrane protein 210:227 a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen 210:335 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 1 43 theme membrane 212:219 arg1 lipase 184:189 lipoprotein lipase 172:189 lipoprotein lipase (LPL) 172:195 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 8 44 with interactions 1401:1412 arg1 GPIHBP1 1419:1425 GPIHBP1 1419:1425 GPIHBP1 1419:1425 In wild-type mice, we expect that the accumulation of LPL in close proximity to capillaries would increase interactions with GPIHBP1. 29899144 6 45 theme GPIHBP1 1027:1033 arg1 IDR 1020:1022 the acidic IDR 1009:1022 the acidic IDR of GPIHBP1 1009:1033 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 10 46 theme GPIHBP1 1659:1665 arg1 IDR 1652:1654 the negatively charged IDR 1629:1654 the negatively charged IDR of GPIHBP1 1629:1665 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 4 47 theme important 708:716 arg1 properties 718:727 several important properties 700:727 several important properties of GPIHBP1's IDR 700:744 Here, we define several important properties of GPIHBP1's IDR. 29899144 10 48 theme vast 1679:1682 arg1 space 1684:1688 a vast space 1677:1688 a vast space 1677:1688 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 1 49 theme capillary 321:329 arg1 lumen 331:335 the capillary lumen 317:335 the capillary lumen 317:335 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 4 50 theme several 700:706 arg1 properties 718:727 several important properties 700:727 several important properties of GPIHBP1's IDR 700:744 Here, we define several important properties of GPIHBP1's IDR. 29899144 10 51 theme LPL 1715:1717 arg1 capture 1704:1710 capture 1704:1710 capture of LPL by capillary endothelial cells 1704:1748 Finally, based on biophysical studies, we propose that the negatively charged IDR of GPIHBP1 traverses a vast space, facilitating capture of LPL by capillary endothelial cells and simultaneously contributing to GPIHBP1's ability to preserve LPL structure and activity. 29899144 8 52 theme close 1355:1359 arg1 proximity 1361:1369 close proximity 1355:1369 close proximity to capillaries 1355:1384 In wild-type mice, we expect that the accumulation of LPL in close proximity to capillaries would increase interactions with GPIHBP1. 29899144 1 53 theme triglyceride-rich 130:146 arg1 lipoproteins 148:159 triglyceride-rich lipoproteins 130:159 triglyceride-rich lipoproteins 130:159 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 3 54 from rich 554:557 arg1 residues 569:576 acidic residues 562:576 acidic residues 562:576 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 9 55 theme chylomicronemia 1508:1522 arg1 pathogenicity 1491:1503 the pathogenicity 1487:1503 the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome 1487:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 8 56 theme wild-type 1297:1305 arg1 mice 1307:1310 wild-type mice 1297:1310 wild-type mice 1297:1310 In wild-type mice, we expect that the accumulation of LPL in close proximity to capillaries would increase interactions with GPIHBP1. 29899144 4 57 theme IDR 742:744 arg1 properties 718:727 several important properties 700:727 several important properties of GPIHBP1's IDR 700:744 Here, we define several important properties of GPIHBP1's IDR. 29899144 1 58 theme lipoproteins 148:159 arg1 processing 116:125 The intravascular processing 98:125 The intravascular processing of triglyceride-rich lipoproteins 98:159 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 6 59 theme rate 1143:1146 arg1 kon 1158:1160 kon 1158:1160 kon 1158:1160 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 6 59 theme rate 1143:1146 arg1 constant 1148:1155 the association rate constant 1127:1155 the association rate constant (kon) for LPL binding 1127:1177 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 3 60 theme ANGPTL4-catalyzed 655:671 arg1 unfolding 673:681 spontaneous and ANGPTL4-catalyzed unfolding 639:681 spontaneous and ANGPTL4-catalyzed unfolding 639:681 The N-terminal domain of GPIHBP1, an intrinsically disordered region (IDR) rich in acidic residues, is important for stabilizing LPL's catalytic domain against spontaneous and ANGPTL4-catalyzed unfolding. 29899144 7 61 dep GPIHBP1 1285:1291 arg1 absence 1274:1280 absence 1274:1280 absence 1274:1280 Third, we show that LPL accumulates near capillary endothelial cells even in the absence of GPIHBP1. 29899144 7 61 dep GPIHBP1 1285:1291 arg1 the 1270:1272 the 1270:1272 the 1270:1272 Third, we show that LPL accumulates near capillary endothelial cells even in the absence of GPIHBP1. 29899144 7 62 theme capillary 1234:1242 arg1 cells 1256:1260 capillary endothelial cells 1234:1260 capillary endothelial cells 1234:1260 Third, we show that LPL accumulates near capillary endothelial cells even in the absence of GPIHBP1. 29899144 6 63 theme LPL 1167:1169 arg1 binding 1171:1177 LPL binding 1167:1177 LPL binding 1167:1177 Second, the acidic IDR of GPIHBP1 increases the probability of a GPIHBP1-LPL encounter via electrostatic steering, increasing the association rate constant (kon) for LPL binding by >250-fold. 29899144 1 64 theme endothelial 232:242 arg1 cells 244:248 endothelial cells 232:248 endothelial cells 232:248 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 0 65 theme sulfated 50:57 arg1 tyrosine 59:66 a sulfated tyrosine 48:66 a sulfated tyrosine 48:66 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine regulates lipoprotein lipase. 29899144 0 66 from domain 20:25 arg1 GPIHBP1 30:36 GPIHBP1 30:36 GPIHBP1 30:36 A disordered acidic domain in GPIHBP1 harboring a sulfated tyrosine regulates lipoprotein lipase. 29899144 9 67 with patients 1527:1534 arg1 syndrome 1564:1571 the GPIHBP1 autoimmune syndrome 1541:1571 the GPIHBP1 autoimmune syndrome 1541:1571 Fourth, we found that GPIHBP1's IDR is not a key factor in the pathogenicity of chylomicronemia in patients with the GPIHBP1 autoimmune syndrome. 29899144 7 68 theme endothelial 1244:1254 arg1 cells 1256:1260 capillary endothelial cells 1234:1260 capillary endothelial cells 1234:1260 Third, we show that LPL accumulates near capillary endothelial cells even in the absence of GPIHBP1. 29899144 1 69 theme cells 244:248 arg1 protein 221:227 a membrane protein 210:227 a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen 210:335 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 29899144 1 69 theme cells 244:248 arg1 lipase 184:189 lipoprotein lipase 172:189 lipoprotein lipase (LPL) 172:195 The intravascular processing of triglyceride-rich lipoproteins depends on lipoprotein lipase (LPL) and GPIHBP1, a membrane protein of endothelial cells that binds LPL within the subendothelial spaces and shuttles it to the capillary lumen. 15907468 8 0 theme acid 1083:1086 arg1 intermediate 1088:1099 a cysteine sulfenic acid intermediate 1063:1099 a cysteine sulfenic acid intermediate 1063:1099 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 7 1 theme redox 920:924 arg1 environments 926:937 different redox environments 910:937 different redox environments 910:937 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 3 2 theme higher 318:323 arg1 eukaryotes 325:334 higher eukaryotes 318:334 higher eukaryotes 318:334 In higher eukaryotes, FGly is generated from a cysteine precursor by the FGly-generating enzyme (FGE). 15907468 5 3 theme surprising 607:616 arg1 paucity 618:624 a surprising paucity 605:624 a surprising paucity of secondary structure 605:647 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 6 4 theme FGE 757:759 arg1 structure 761:769 the FGE structure 753:769 the FGE structure 753:769 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 6 5 theme MSD 803:805 arg1 basis 794:798 a molecular basis 782:798 a molecular basis of MSD 782:805 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 0 6 theme human 100:104 arg1 enzyme 131:136 the human formylglycine-generating enzyme 96:136 the human formylglycine-generating enzyme 96:136 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 8 7 theme oxygenase 984:992 arg1 mechanism 994:1002 a novel oxygenase mechanism 976:1002 a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate 976:1099 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 4 8 theme sulfatase 456:464 arg1 deficiency 466:475 multiple sulfatase deficiency 447:475 multiple sulfatase deficiency (MSD) 447:481 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 8 theme sulfatase 456:464 arg1 syndrome 512:519 a fatal autosomal recessive syndrome 484:519 a fatal autosomal recessive syndrome 484:519 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 8 theme sulfatase 456:464 arg1 MSD 478:480 MSD 478:480 MSD 478:480 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 8 9 theme novel 978:982 arg1 mechanism 994:1002 a novel oxygenase mechanism 976:1002 a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate 976:1099 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 8 10 theme molecular 1025:1033 arg1 oxygen 1035:1040 molecular oxygen 1025:1040 molecular oxygen 1025:1040 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 0 11 theme enzyme 131:136 arg1 generation 82:91 formylglycine generation 68:91 formylglycine generation of the human formylglycine-generating enzyme 68:136 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 4 12 theme multiple 447:454 arg1 deficiency 466:475 multiple sulfatase deficiency 447:475 multiple sulfatase deficiency (MSD) 447:481 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 12 theme multiple 447:454 arg1 syndrome 512:519 a fatal autosomal recessive syndrome 484:519 a fatal autosomal recessive syndrome 484:519 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 12 theme multiple 447:454 arg1 MSD 478:480 MSD 478:480 MSD 478:480 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 0 13 theme formylglycine-generating 106:129 arg1 enzyme 131:136 the human formylglycine-generating enzyme 96:136 the human formylglycine-generating enzyme 96:136 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 5 14 theme single-domain 578:590 arg1 monomer 592:598 a single-domain monomer 576:598 a single-domain monomer with a surprising paucity of secondary structure 576:647 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 5 14 theme single-domain 578:590 arg1 FGE 569:571 FGE 569:571 FGE 569:571 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 5 15 theme crystal 535:541 arg1 structure 543:551 the crystal structure 531:551 the crystal structure 531:551 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 2 16 theme active 277:282 arg1 site 284:287 the active site 273:287 the active site 273:287 Formylglycine (FGly), the key catalytic residue in the active site, is unique to sulfatases. 15907468 7 17 theme high-resolution 873:887 arg1 structures 889:898 six high-resolution structures 869:898 six high-resolution structures of FGE in different redox environments 869:937 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 1 18 theme sulfate 206:212 arg1 esters 214:219 sulfate esters 206:219 sulfate esters 206:219 Sulfatases are enzymes essential for degradation and remodeling of sulfate esters. 15907468 0 19 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis for multiple sulfatase deficiency 0:48 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 6 20 theme MSD 724:726 arg1 patients 728:735 MSD patients 724:735 MSD patients 724:735 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 2 21 from residue 262:268 arg1 site 284:287 the active site 273:287 the active site 273:287 Formylglycine (FGly), the key catalytic residue in the active site, is unique to sulfatases. 15907468 1 22 theme esters 214:219 arg1 remodeling 192:201 remodeling 192:201 remodeling 192:201 Sulfatases are enzymes essential for degradation and remodeling of sulfate esters. 15907468 1 22 theme esters 214:219 arg1 degradation 176:186 degradation 176:186 degradation 176:186 Sulfatases are enzymes essential for degradation and remodeling of sulfate esters. 15907468 0 23 theme multiple 20:27 arg1 deficiency 39:48 multiple sulfatase deficiency 20:48 multiple sulfatase deficiency 20:48 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 7 24 theme FGE 903:905 arg1 structures 889:898 six high-resolution structures 869:898 six high-resolution structures of FGE in different redox environments 869:937 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 3 25 theme cysteine 362:369 arg1 precursor 371:379 a cysteine precursor 360:379 a cysteine precursor 360:379 In higher eukaryotes, FGly is generated from a cysteine precursor by the FGly-generating enzyme (FGE). 15907468 5 26 theme unique 662:667 arg1 fold 669:672 a unique fold 660:672 a unique fold 660:672 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 4 27 theme FGE 432:434 arg1 Inactivity 418:427 Inactivity 418:427 Inactivity of FGE 418:434 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 7 28 from structures 889:898 arg1 environments 926:937 different redox environments 910:937 different redox environments 910:937 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 2 29 theme key 248:250 arg1 residue 262:268 the key catalytic residue 244:268 the key catalytic residue in the active site 244:287 Formylglycine (FGly), the key catalytic residue in the active site, is unique to sulfatases. 15907468 2 29 theme key 248:250 arg1 Formylglycine 222:234 Formylglycine 222:234 Formylglycine (FGly) 222:241 Formylglycine (FGly), the key catalytic residue in the active site, is unique to sulfatases. 15907468 0 30 theme sulfatase 29:37 arg1 deficiency 39:48 multiple sulfatase deficiency 20:48 multiple sulfatase deficiency 20:48 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 4 31 theme autosomal 492:500 arg1 deficiency 466:475 multiple sulfatase deficiency 447:475 multiple sulfatase deficiency (MSD) 447:481 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 31 theme autosomal 492:500 arg1 syndrome 512:519 a fatal autosomal recessive syndrome 484:519 a fatal autosomal recessive syndrome 484:519 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 6 32 located found 715:719 arg1 patients 728:735 MSD patients 724:735 MSD patients 724:735 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 6 32 located found 715:719 arg2 mutations 705:713 all 18 missense mutations 689:713 all 18 missense mutations found in MSD patients 689:735 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 4 33 theme fatal 486:490 arg1 deficiency 466:475 multiple sulfatase deficiency 447:475 multiple sulfatase deficiency (MSD) 447:481 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 33 theme fatal 486:490 arg1 syndrome 512:519 a fatal autosomal recessive syndrome 484:519 a fatal autosomal recessive syndrome 484:519 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 5 34 theme secondary 629:637 arg1 structure 639:647 secondary structure 629:647 secondary structure 629:647 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 6 35 theme molecular 784:792 arg1 basis 794:798 a molecular basis 782:798 a molecular basis of MSD 782:805 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 6 36 theme mutations 705:713 arg1 effect 679:684 The effect 675:684 The effect of all 18 missense mutations found in MSD patients 675:735 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 5 37 with monomer 592:598 arg1 paucity 618:624 a surprising paucity 605:624 a surprising paucity of secondary structure 605:647 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 3 38 theme FGly-generating 388:402 arg1 FGE 412:414 FGE 412:414 FGE 412:414 In higher eukaryotes, FGly is generated from a cysteine precursor by the FGly-generating enzyme (FGE). 15907468 3 38 theme FGly-generating 388:402 arg1 enzyme 404:409 the FGly-generating enzyme 384:409 the FGly-generating enzyme (FGE) 384:415 In higher eukaryotes, FGly is generated from a cysteine precursor by the FGly-generating enzyme (FGE). 15907468 5 39 theme structure 639:647 arg1 paucity 618:624 a surprising paucity 605:624 a surprising paucity of secondary structure 605:647 Based on the crystal structure, we report that FGE is a single-domain monomer with a surprising paucity of secondary structure and adopts a unique fold. 15907468 8 40 theme mechanism 994:1002 arg1 formulation 961:971 formulation 961:971 formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate 961:1099 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 6 41 theme missense 696:703 arg1 mutations 705:713 all 18 missense mutations 689:713 all 18 missense mutations found in MSD patients 689:735 The effect of all 18 missense mutations found in MSD patients is explained by the FGE structure, providing a molecular basis of MSD. 15907468 7 42 theme generation 840:849 arg1 mechanism 822:830 The catalytic mechanism 808:830 The catalytic mechanism of FGly generation 808:849 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 4 43 theme recessive 502:510 arg1 deficiency 466:475 multiple sulfatase deficiency 447:475 multiple sulfatase deficiency (MSD) 447:481 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 4 43 theme recessive 502:510 arg1 syndrome 512:519 a fatal autosomal recessive syndrome 484:519 a fatal autosomal recessive syndrome 484:519 Inactivity of FGE results in multiple sulfatase deficiency (MSD), a fatal autosomal recessive syndrome. 15907468 8 44 theme cysteine 1065:1072 arg1 intermediate 1088:1099 a cysteine sulfenic acid intermediate 1063:1099 a cysteine sulfenic acid intermediate 1063:1099 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 0 45 theme formylglycine 68:80 arg1 generation 82:91 formylglycine generation 68:91 formylglycine generation of the human formylglycine-generating enzyme 68:136 Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. 15907468 1 46 theme essential 162:170 arg1 enzymes 154:160 enzymes 154:160 enzymes essential for degradation and remodeling of sulfate esters 154:219 Sulfatases are enzymes essential for degradation and remodeling of sulfate esters. 15907468 1 46 theme essential 162:170 arg1 Sulfatases 139:148 Sulfatases 139:148 Sulfatases 139:148 Sulfatases are enzymes essential for degradation and remodeling of sulfate esters. 15907468 7 47 theme FGly 835:838 arg1 generation 840:849 FGly generation 835:849 FGly generation 835:849 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 8 48 theme sulfenic 1074:1081 arg1 intermediate 1088:1099 a cysteine sulfenic acid intermediate 1063:1099 a cysteine sulfenic acid intermediate 1063:1099 The structures allow formulation of a novel oxygenase mechanism whereby FGE utilizes molecular oxygen to generate FGly via a cysteine sulfenic acid intermediate. 15907468 2 49 theme catalytic 252:260 arg1 residue 262:268 the key catalytic residue 244:268 the key catalytic residue in the active site 244:287 Formylglycine (FGly), the key catalytic residue in the active site, is unique to sulfatases. 15907468 2 49 theme catalytic 252:260 arg1 Formylglycine 222:234 Formylglycine 222:234 Formylglycine (FGly) 222:241 Formylglycine (FGly), the key catalytic residue in the active site, is unique to sulfatases. 15907468 7 50 theme different 910:918 arg1 environments 926:937 different redox environments 910:937 different redox environments 910:937 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 15907468 7 51 theme catalytic 812:820 arg1 mechanism 822:830 The catalytic mechanism 808:830 The catalytic mechanism of FGly generation 808:849 The catalytic mechanism of FGly generation was elucidated by six high-resolution structures of FGE in different redox environments. 22940367 9 0 theme diseases 1596:1603 arg1 family 1586:1591 the larger MPS family 1571:1591 the larger MPS family of diseases 1571:1603 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 1 1 theme inherited 223:231 arg1 defect 243:248 an inherited metabolic defect 220:248 an inherited metabolic defect known as a lysosomal storage disorder 220:286 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 8 2 theme paralogous 1338:1347 arg1 sulfatases 1349:1358 paralogous sulfatases 1338:1358 paralogous sulfatases 1338:1358 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 7 3 theme A 1106:1106 arg1 mutations 1117:1125 120 MPS IV A missense mutations 1095:1125 120 MPS IV A missense mutations onto the structure 1095:1144 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 7 4 theme MPS 1099:1101 arg1 mutations 1117:1125 120 MPS IV A missense mutations 1095:1125 120 MPS IV A missense mutations onto the structure 1095:1144 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 4 5 attach derived 726:732 arg1 modification 739:750 modification 739:750 modification of a cysteine side chain 739:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 4 5 attach derived 726:732 arg2 nucleophile 714:724 a catalytic gem diol nucleophile 693:724 a catalytic gem diol nucleophile derived from modification of a cysteine side chain 693:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 8 6 from geometries 1396:1405 arg1 family 1414:1419 the family 1410:1419 the family 1410:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 5 7 theme GALNS 797:801 arg1 trench 834:839 a large, positively charged trench 806:839 a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate 806:933 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 7 theme GALNS 797:801 arg1 GALNS 797:801 GALNS 797:801 GALNS 797:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 7 theme GALNS 797:801 arg1 site 789:792 The active site 778:792 The active site of GALNS 778:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 2 8 theme MPS 526:528 arg1 A 533:533 MPS IV A 526:533 MPS IV A 526:533 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 5 9 theme active 782:787 arg1 trench 834:839 a large, positively charged trench 806:839 a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate 806:933 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 9 theme active 782:787 arg1 GALNS 797:801 GALNS 797:801 GALNS 797:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 9 theme active 782:787 arg1 site 789:792 The active site 778:792 The active site of GALNS 778:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 8 10 theme strict 1425:1430 arg1 conservation 1432:1443 strict conservation 1425:1443 strict conservation of the catalytic machinery 1425:1470 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 2 11 theme Morquio 539:545 arg1 A 547:547 Morquio A 539:547 Morquio A 539:547 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 12 theme IV 506:507 arg1 A 509:509 mucopolysaccharidosis IV A 484:509 the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 454:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 8 13 theme catalytic 1452:1460 arg1 machinery 1462:1470 the catalytic machinery 1448:1470 the catalytic machinery 1448:1470 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 7 14 theme structure 1218:1226 arg1 core 1206:1209 the hydrophobic core 1190:1209 the hydrophobic core of the structure 1190:1226 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 5 15 theme suitable 841:848 arg1 trench 834:839 a large, positively charged trench 806:839 a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate 806:933 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 15 theme suitable 841:848 arg1 GALNS 797:801 GALNS 797:801 GALNS 797:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 15 theme suitable 841:848 arg1 site 789:792 The active site 778:792 The active site of GALNS 778:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 8 16 from family 1414:1419 arg1 variety 1373:1379 a wide variety 1366:1379 a wide variety of active-site geometries in the family 1366:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 8 16 from family 1414:1419 arg1 geometries 1396:1405 active-site geometries 1384:1405 active-site geometries in the family 1384:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 9 17 theme known 1504:1508 arg1 mutations 1510:1518 the known mutations 1500:1518 the known mutations 1500:1518 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 6 18 theme human 982:986 arg1 GALNS 988:992 the insect-cell-expressed human GALNS 956:992 the insect-cell-expressed human GALNS 956:992 Enzymatic assays on the insect-cell-expressed human GALNS indicate activity against synthetic substrates and inhibition by both substrate and product. 22940367 2 19 from deficient 427:435 arg1 patients 440:447 patients 440:447 patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 440:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 20 dep GALNS 343:347 arg1 E.C. 410:413 E.C. 410:413 E.C. 410:413 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 0 21 dep basis 51:55 arg1 catalyze 107:114 catalyze 107:114 catalyze the breakdown of macromolecules in the cell 107:158 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 0 22 theme A. 86:87 arg1 enzymes 99:105 A. Lysosomal enzymes 86:105 A. Lysosomal enzymes 86:105 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 2 23 from patients 440:447 arg1 deficient 427:435 deficient 427:435 deficient 427:435 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 23 from patients 440:447 arg1 galactosamine-6-sulfatase 316:340 The human lysosomal enzyme galactosamine-6-sulfatase 289:340 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) 289:422 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 8 24 theme GALNS 1329:1333 arg1 structure 1316:1324 the structure 1312:1324 the structure of GALNS 1312:1333 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 9 25 theme molecular 1534:1542 arg1 basis 1544:1548 the molecular basis 1530:1548 the molecular basis for MPS IV A and for the larger MPS family of diseases 1530:1603 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 4 26 theme gem 705:707 arg1 nucleophile 714:724 a catalytic gem diol nucleophile 693:724 a catalytic gem diol nucleophile derived from modification of a cysteine side chain 693:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 2 27 dep galactosamine-6-sulfatase 316:340 arg1 GALNS 343:347 GALNS 343:347 GALNS 343:347 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 9 28 theme IV 1558:1559 arg1 A 1561:1561 MPS IV A 1554:1561 MPS IV A 1554:1561 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 2 29 theme lysosomal 458:466 arg1 disease 476:482 the lysosomal storage disease 454:482 the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 454:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 8 30 theme wide 1368:1371 arg1 variety 1373:1379 a wide variety 1366:1379 a wide variety of active-site geometries in the family 1366:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 8 30 theme wide 1368:1371 arg1 geometries 1396:1405 active-site geometries 1384:1405 active-site geometries in the family 1384:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 0 31 theme GALNS 23:27 arg1 structure 4:12 The structure 0:12 The structure of human GALNS 0:27 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 1 32 theme storage 271:277 arg1 disorder 279:286 a lysosomal storage disorder 259:286 a lysosomal storage disorder 259:286 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 3 33 theme human 602:606 arg1 GALNS 608:612 human GALNS 602:612 human GALNS 602:612 Here, we report the three-dimensional structure of human GALNS, determined by X-ray crystallography at 2.2Å resolution. 22940367 5 34 dep large 808:812 arg1 charged 826:832 charged 826:832 charged 826:832 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 8 35 theme geometries 1396:1405 arg1 variety 1373:1379 a wide variety 1366:1379 a wide variety of active-site geometries in the family 1366:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 8 35 theme geometries 1396:1405 arg1 geometries 1396:1405 active-site geometries 1384:1405 active-site geometries in the family 1384:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 2 36 theme lysosomal 299:307 arg1 deficient 427:435 deficient 427:435 deficient 427:435 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 36 theme lysosomal 299:307 arg1 galactosamine-6-sulfatase 316:340 The human lysosomal enzyme galactosamine-6-sulfatase 289:340 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) 289:422 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 1 37 theme lysosomal 194:202 arg1 enzyme 204:209 a lysosomal enzyme 192:209 a lysosomal enzyme 192:209 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 9 38 theme MPS 1582:1584 arg1 family 1586:1591 the larger MPS family 1571:1591 the larger MPS family of diseases 1571:1603 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 7 39 theme IV 1254:1255 arg1 cases 1259:1263 most MPS IV A cases 1245:1263 most MPS IV A cases 1245:1263 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 4 40 theme chain 771:775 arg1 modification 739:750 modification 739:750 modification of a cysteine side chain 739:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 7 41 theme most 1245:1248 arg1 cases 1259:1263 most MPS IV A cases 1245:1263 most MPS IV A cases 1245:1263 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 7 42 theme missense 1108:1115 arg1 mutations 1117:1125 120 MPS IV A missense mutations 1095:1125 120 MPS IV A missense mutations onto the structure 1095:1144 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 3 43 theme 2.2Å 654:657 arg1 resolution 659:668 2.2Å resolution 654:668 2.2Å resolution 654:668 Here, we report the three-dimensional structure of human GALNS, determined by X-ray crystallography at 2.2Å resolution. 22940367 7 44 theme IV 1103:1104 arg1 mutations 1117:1125 120 MPS IV A missense mutations 1095:1125 120 MPS IV A missense mutations onto the structure 1095:1144 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 1 45 theme metabolic 233:241 arg1 defect 243:248 an inherited metabolic defect 220:248 an inherited metabolic defect known as a lysosomal storage disorder 220:286 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 8 46 from variety 1373:1379 arg1 family 1414:1419 the family 1410:1419 the family 1410:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 5 47 theme large 808:812 arg1 trench 834:839 a large, positively charged trench 806:839 a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate 806:933 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 47 theme large 808:812 arg1 GALNS 797:801 GALNS 797:801 GALNS 797:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 47 theme large 808:812 arg1 site 789:792 The active site 778:792 The active site of GALNS 778:801 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 6 48 theme Enzymatic 936:944 arg1 assays 946:951 Enzymatic assays 936:951 Enzymatic assays on the insect-cell-expressed human GALNS 936:992 Enzymatic assays on the insect-cell-expressed human GALNS indicate activity against synthetic substrates and inhibition by both substrate and product. 22940367 2 49 theme IV 530:531 arg1 A 533:533 MPS IV A 526:533 MPS IV A 526:533 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 0 50 theme macromolecules 133:146 arg1 breakdown 120:128 the breakdown 116:128 the breakdown of macromolecules in the cell 116:158 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 3 51 from resolution 659:668 arg1 crystallography 635:649 X-ray crystallography 629:649 X-ray crystallography at 2.2Å resolution 629:668 Here, we report the three-dimensional structure of human GALNS, determined by X-ray crystallography at 2.2Å resolution. 22940367 5 52 theme polyanionic 862:872 arg1 substrates 874:883 polyanionic substrates 862:883 polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate 862:933 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 52 theme polyanionic 862:872 arg1 chondroitin-6-sulfate 913:933 chondroitin-6-sulfate 913:933 chondroitin-6-sulfate 913:933 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 5 52 theme polyanionic 862:872 arg1 sulfate 901:907 keratan sulfate 893:907 keratan sulfate 893:907 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 4 53 theme cysteine 757:764 arg1 chain 771:775 a cysteine side chain 755:775 a cysteine side chain 755:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 3 54 theme three-dimensional 571:587 arg1 structure 589:597 the three-dimensional structure 567:597 the three-dimensional structure 567:597 Here, we report the three-dimensional structure of human GALNS, determined by X-ray crystallography at 2.2Å resolution. 22940367 6 55 theme synthetic 1020:1028 arg1 substrates 1030:1039 synthetic substrates 1020:1039 synthetic substrates 1020:1039 Enzymatic assays on the insect-cell-expressed human GALNS indicate activity against synthetic substrates and inhibition by both substrate and product. 22940367 8 56 theme machinery 1462:1470 arg1 conservation 1432:1443 strict conservation 1425:1443 strict conservation of the catalytic machinery 1425:1470 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 8 57 theme structure 1316:1324 arg1 Comparison 1298:1307 Comparison 1298:1307 Comparison of the structure of GALNS to paralogous sulfatases 1298:1358 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 2 58 with patients 440:447 arg1 disease 476:482 the lysosomal storage disease 454:482 the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 454:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 59 theme mucopolysaccharidosis 484:504 arg1 A 509:509 mucopolysaccharidosis IV A 484:509 the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 454:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 7 60 theme hydrophobic 1194:1204 arg1 core 1206:1209 the hydrophobic core 1190:1209 the hydrophobic core of the structure 1190:1226 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 2 61 theme storage 468:474 arg1 disease 476:482 the lysosomal storage disease 454:482 the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 454:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 0 62 theme Lysosomal 89:97 arg1 enzymes 99:105 A. Lysosomal enzymes 86:105 A. Lysosomal enzymes 86:105 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 6 63 theme insect-cell-expressed 960:980 arg1 GALNS 988:992 the insect-cell-expressed human GALNS 956:992 the insect-cell-expressed human GALNS 956:992 Enzymatic assays on the insect-cell-expressed human GALNS indicate activity against synthetic substrates and inhibition by both substrate and product. 22940367 7 64 theme mutations 1173:1181 arg1 majority 1161:1168 a majority 1159:1168 a majority of mutations 1159:1181 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 1 65 theme activity 180:187 arg1 loss 172:175 loss 172:175 loss of activity of a lysosomal enzyme 172:209 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 5 66 theme keratan 893:899 arg1 sulfate 901:907 keratan sulfate 893:907 keratan sulfate 893:907 The active site of GALNS is a large, positively charged trench suitable for binding polyanionic substrates such as keratan sulfate and chondroitin-6-sulfate. 22940367 7 67 theme GALNS 1291:1295 arg1 misfolding 1277:1286 misfolding 1277:1286 misfolding of GALNS 1277:1295 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 4 68 theme catalytic 695:703 arg1 nucleophile 714:724 a catalytic gem diol nucleophile 693:724 a catalytic gem diol nucleophile derived from modification of a cysteine side chain 693:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 0 69 theme human 17:21 arg1 GALNS 23:27 human GALNS 17:27 human GALNS 17:27 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 9 70 theme MPS 1554:1556 arg1 A 1561:1561 MPS IV A 1554:1561 MPS IV A 1554:1561 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 3 71 theme GALNS 608:612 arg1 structure 589:597 the three-dimensional structure 567:597 the three-dimensional structure 567:597 Here, we report the three-dimensional structure of human GALNS, determined by X-ray crystallography at 2.2Å resolution. 22940367 4 72 theme diol 709:712 arg1 nucleophile 714:724 a catalytic gem diol nucleophile 693:724 a catalytic gem diol nucleophile derived from modification of a cysteine side chain 693:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 22940367 1 73 theme lysosomal 261:269 arg1 disorder 279:286 a lysosomal storage disorder 259:286 a lysosomal storage disorder 259:286 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 2 74 dep known 517:521 arg1 also 512:515 also 512:515 also 512:515 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 6 75 from assays 946:951 arg1 GALNS 988:992 the insect-cell-expressed human GALNS 956:992 the insect-cell-expressed human GALNS 956:992 Enzymatic assays on the insect-cell-expressed human GALNS indicate activity against synthetic substrates and inhibition by both substrate and product. 22940367 0 76 theme molecular 41:49 arg1 basis 51:55 the molecular basis 37:55 the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell 37:158 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 2 77 dep disease 476:482 arg1 A 509:509 mucopolysaccharidosis IV A 484:509 the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A) 454:548 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 77 dep disease 476:482 arg1 known 517:521 known 517:521 known as MPS IV A and Morquio A 517:547 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 7 78 theme A 1257:1257 arg1 cases 1259:1263 most MPS IV A cases 1245:1263 most MPS IV A cases 1245:1263 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 3 79 theme X-ray 629:633 arg1 crystallography 635:649 X-ray crystallography 629:649 X-ray crystallography at 2.2Å resolution 629:668 Here, we report the three-dimensional structure of human GALNS, determined by X-ray crystallography at 2.2Å resolution. 22940367 2 80 theme enzyme 309:314 arg1 deficient 427:435 deficient 427:435 deficient 427:435 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 80 theme enzyme 309:314 arg1 galactosamine-6-sulfatase 316:340 The human lysosomal enzyme galactosamine-6-sulfatase 289:340 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) 289:422 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 9 81 theme larger 1575:1580 arg1 family 1586:1591 the larger MPS family 1571:1591 the larger MPS family of diseases 1571:1603 Overall, the structure and the known mutations establish the molecular basis for MPS IV A and for the larger MPS family of diseases. 22940367 7 82 theme MPS 1250:1252 arg1 cases 1259:1263 most MPS IV A cases 1245:1263 most MPS IV A cases 1245:1263 Mapping 120 MPS IV A missense mutations onto the structure reveals that a majority of mutations affect the hydrophobic core of the structure, indicating that most MPS IV A cases result from misfolding of GALNS. 22940367 8 83 theme active-site 1384:1394 arg1 geometries 1396:1405 active-site geometries 1384:1405 active-site geometries in the family 1384:1419 Comparison of the structure of GALNS to paralogous sulfatases shows a wide variety of active-site geometries in the family but strict conservation of the catalytic machinery. 22940367 2 84 theme human 293:297 arg1 deficient 427:435 deficient 427:435 deficient 427:435 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 2 84 theme human 293:297 arg1 galactosamine-6-sulfatase 316:340 The human lysosomal enzyme galactosamine-6-sulfatase 289:340 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) 289:422 The human lysosomal enzyme galactosamine-6-sulfatase (GALNS, also known as N-acetylgalactosamine-6-sulfatase and GalN6S; E.C. 3.1.6.4) is deficient in patients with the lysosomal storage disease mucopolysaccharidosis IV A (also known as MPS IV A and Morquio A). 22940367 1 85 theme enzyme 204:209 arg1 activity 180:187 activity 180:187 activity of a lysosomal enzyme 180:209 In humans, loss of activity of a lysosomal enzyme leads to an inherited metabolic defect known as a lysosomal storage disorder. 22940367 0 86 from breakdown 120:128 arg1 cell 155:158 the cell 151:158 the cell 151:158 The structure of human GALNS reveals the molecular basis for mucopolysaccharidosis IV A. Lysosomal enzymes catalyze the breakdown of macromolecules in the cell. 22940367 4 87 theme side 766:769 arg1 chain 771:775 a cysteine side chain 755:775 a cysteine side chain 755:775 The structure reveals a catalytic gem diol nucleophile derived from modification of a cysteine side chain. 19478429 2 0 theme PDB 383:385 arg1 2v5e 392:395 PDB code 2v5e 383:395 PDB code 2v5e 383:395 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 2 0 theme PDB 383:385 arg1 complex 374:380 the GDNF(2)-GFRalpha1(2) complex 349:380 the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) 349:396 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 5 1 theme small 944:948 arg1 change 950:955 a small change 942:955 a small change in the structure of the GDNF 942:984 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 6 2 theme structures 1031:1040 arg1 comparison 990:999 comparison 990:999 comparison of all known GDNF and artemin structures 990:1040 By comparison of all known GDNF and artemin structures, it is concluded that GDNF is more bent and more flexible than artemin and that this may be related to RET signalling. 19478429 2 3 theme ligand 454:459 arg1 complexes 467:475 GDNF-family ligand (GFL) complexes 442:475 GDNF-family ligand (GFL) complexes 442:475 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 7 4 theme monomer-monomer 1345:1359 arg1 interface 1361:1369 the monomer-monomer interface 1341:1369 the monomer-monomer interface 1341:1369 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 3 5 theme new 661:663 arg1 dimensions 670:679 new cell dimensions 661:679 new cell dimensions 661:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 1 6 theme -RET 286:289 arg1 complex 308:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 6 7 theme artemin 1023:1029 arg1 structures 1031:1040 all known GDNF and artemin structures 1004:1040 all known GDNF and artemin structures 1004:1040 By comparison of all known GDNF and artemin structures, it is concluded that GDNF is more bent and more flexible than artemin and that this may be related to RET signalling. 19478429 3 8 theme cell 665:668 arg1 dimensions 670:679 new cell dimensions 661:679 new cell dimensions 661:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 8 9 theme artemin-like 1495:1506 arg1 conformation 1508:1519 an artemin-like conformation 1492:1519 an artemin-like conformation 1492:1519 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 8 10 theme sequence 1417:1424 arg1 comparison 1426:1435 sequence comparison 1417:1435 sequence comparison 1417:1435 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 0 11 theme RET 82:84 arg1 signalling 86:95 RET signalling 82:95 RET signalling 82:95 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 3 12 theme A 584:584 arg1 structure 597:605 a 2.35 A resolution structure 577:605 a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions 577:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 6 13 theme GDNF 1014:1017 arg1 structures 1031:1040 all known GDNF and artemin structures 1004:1040 all known GDNF and artemin structures 1004:1040 By comparison of all known GDNF and artemin structures, it is concluded that GDNF is more bent and more flexible than artemin and that this may be related to RET signalling. 19478429 4 14 theme final 725:729 arg1 factor 733:738 a final R factor 723:738 a final R factor of 22.5% (R(free) = 28%) 723:763 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 7 15 theme artemin 1274:1280 arg1 curvature 1257:1265 the increased curvature 1243:1265 the increased curvature of the artemin 1243:1280 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 3 16 theme resolution 586:595 arg1 structure 597:605 a 2.35 A resolution structure 577:605 a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions 577:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 2 17 theme bend 513:516 arg1 angle 518:522 the bend angle 509:522 the bend angle between the two monomers in the GFL homodimer 509:568 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 5 18 from structures 770:779 arg1 unit 839:842 the asymmetric unit 824:842 the asymmetric unit 824:842 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 2 19 theme crystal 328:334 arg1 structure 336:344 A previous crystal structure 317:344 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) 317:396 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 4 20 dep factor 733:738 arg1 R 750:750 R 750:750 R(free) = 28% 750:762 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 1 21 theme heterohexamer 294:306 arg1 complex 308:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 2 22 theme complex 374:380 arg1 structure 336:344 A previous crystal structure 317:344 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) 317:396 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 2 23 theme GFL 556:558 arg1 homodimer 560:568 the GFL homodimer 552:568 the GFL homodimer 552:568 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 3 24 theme complex 635:641 arg1 structure 597:605 a 2.35 A resolution structure 577:605 a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions 577:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 7 25 theme intermonomer 1387:1398 arg1 angle 1405:1409 the intermonomer bend angle 1383:1409 the intermonomer bend angle 1383:1409 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 1 26 theme receptor 163:170 arg1 RET 188:190 the receptor tyrosine kinase RET 159:190 the receptor tyrosine kinase RET 159:190 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 2 27 theme -GFRalpha1 360:369 arg1 2v5e 392:395 PDB code 2v5e 383:395 PDB code 2v5e 383:395 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 2 27 theme -GFRalpha1 360:369 arg1 complex 374:380 the GDNF(2)-GFRalpha1(2) complex 349:380 the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) 349:396 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 3 28 theme 2.35 579:582 arg1 structure 597:605 a 2.35 A resolution structure 577:605 a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions 577:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 4 29 theme = 758:758 arg1 R 750:750 R 750:750 R(free) = 28% 750:762 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 7 30 theme bend 1400:1403 arg1 angle 1405:1409 the intermonomer bend angle 1383:1409 the intermonomer bend angle 1383:1409 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 1 31 theme tyrosine 172:179 arg1 RET 188:190 the receptor tyrosine kinase RET 159:190 the receptor tyrosine kinase RET 159:190 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 4 32 dep R 750:750 arg1 free 752:755 free 752:755 free 752:755 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 8 33 theme different 1544:1552 arg1 conformation 1554:1565 a different conformation 1542:1565 a different conformation to the other three 1542:1584 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 6 34 theme known 1008:1012 arg1 structures 1031:1040 all known GDNF and artemin structures 1004:1040 all known GDNF and artemin structures 1004:1040 By comparison of all known GDNF and artemin structures, it is concluded that GDNF is more bent and more flexible than artemin and that this may be related to RET signalling. 19478429 5 35 theme artemin-GFRalpha3 892:908 arg1 structure 910:918 the artemin-GFRalpha3 structure 888:918 the artemin-GFRalpha3 structure 888:918 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 1 36 theme kinase 181:186 arg1 RET 188:190 the receptor tyrosine kinase RET 159:190 the receptor tyrosine kinase RET 159:190 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 0 37 theme complexes 27:35 arg1 Comparison 0:9 Comparison 0:9 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 0:96 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 3 38 theme GDNF 614:617 arg1 complex 635:641 the GDNF(2)-GFRalpha1(2) complex 610:641 the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions 610:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 19478429 2 39 theme GDNF-family 442:452 arg1 complexes 467:475 GDNF-family ligand (GFL) complexes 442:475 GDNF-family ligand (GFL) complexes 442:475 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 2 40 theme GDNF 353:356 arg1 2v5e 392:395 PDB code 2v5e 383:395 PDB code 2v5e 383:395 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 2 40 theme GDNF 353:356 arg1 complex 374:380 the GDNF(2)-GFRalpha1(2) complex 349:380 the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) 349:396 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 0 41 theme GFL-GFRalpha 14:25 arg1 complexes 27:35 GFL-GFRalpha complexes 14:35 GFL-GFRalpha complexes 14:35 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 1 42 theme GDNF 266:269 arg1 complex 308:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 43 theme Glial 98:102 arg1 GDNF 143:146 GDNF 143:146 GDNF 143:146 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 43 theme Glial 98:102 arg1 factor 135:140 Glial cell line-derived neurotrophic factor 98:140 Glial cell line-derived neurotrophic factor (GDNF) 98:147 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 0 44 theme further 38:44 arg1 evidence 46:53 further evidence 38:53 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 0:96 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 4 45 theme % 747:747 arg1 factor 733:738 a final R factor 723:738 a final R factor of 22.5% (R(free) = 28%) 723:763 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 4 46 dep = 758:758 arg1 % 762:762 28% 760:762 28% 760:762 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 7 47 dep increases 1304:1312 arg1 both 1299:1302 both 1299:1302 both 1299:1302 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 5 48 from change 950:955 arg1 structure 964:972 the structure 960:972 the structure of the GDNF 960:984 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 7 49 theme buried 1318:1323 arg1 area 1333:1336 the buried surface area 1314:1336 the buried surface area in the monomer-monomer interface 1314:1369 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 5 50 from structure 910:918 arg1 different 873:881 different 873:881 different 873:881 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 6 51 theme RET 1145:1147 arg1 signalling 1149:1158 RET signalling 1145:1158 RET signalling 1145:1158 By comparison of all known GDNF and artemin structures, it is concluded that GDNF is more bent and more flexible than artemin and that this may be related to RET signalling. 19478429 2 52 from differences 494:504 arg1 angle 518:522 the bend angle 509:522 the bend angle between the two monomers in the GFL homodimer 509:568 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 7 53 from area 1333:1336 arg1 interface 1361:1369 the monomer-monomer interface 1341:1369 the monomer-monomer interface 1341:1369 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 7 54 theme increased 1247:1255 arg1 curvature 1257:1265 the increased curvature 1243:1265 the increased curvature of the artemin 1243:1280 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 5 55 theme biological 789:798 arg1 complexes 811:819 both biological tetrameric complexes 784:819 both biological tetrameric complexes 784:819 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 4 56 theme R 731:731 arg1 factor 733:738 a final R factor 723:738 a final R factor of 22.5% (R(free) = 28%) 723:763 The structure was refined to a final R factor of 22.5% (R(free) = 28%). 19478429 1 57 theme cell 104:107 arg1 GDNF 143:146 GDNF 143:146 GDNF 143:146 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 57 theme cell 104:107 arg1 factor 135:140 Glial cell line-derived neurotrophic factor 98:140 Glial cell line-derived neurotrophic factor (GDNF) 98:147 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 58 theme -GFRalpha1 273:282 arg1 complex 308:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex 262:314 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 2 59 theme GFL 462:464 arg1 complexes 467:475 GDNF-family ligand (GFL) complexes 442:475 GDNF-family ligand (GFL) complexes 442:475 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 5 60 theme complexes 811:819 arg1 structures 770:779 The structures 766:779 The structures of both biological tetrameric complexes in the asymmetric unit 766:842 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 5 60 theme complexes 811:819 arg1 similar 853:859 similar 853:859 similar 853:859 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 5 61 theme tetrameric 800:809 arg1 complexes 811:819 both biological tetrameric complexes 784:819 both biological tetrameric complexes 784:819 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 8 62 theme second 1473:1478 arg1 neuturin 1459:1466 neuturin 1459:1466 neuturin (the second GFL) 1459:1483 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 8 62 theme second 1473:1478 arg1 GFL 1480:1482 the second GFL 1469:1482 the second GFL 1469:1482 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 1 63 theme line-derived 109:120 arg1 GDNF 143:146 GDNF 143:146 GDNF 143:146 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 63 theme line-derived 109:120 arg1 factor 135:140 Glial cell line-derived neurotrophic factor 98:140 Glial cell line-derived neurotrophic factor (GDNF) 98:147 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 5 64 theme GDNF 981:984 arg1 structure 964:972 the structure 960:972 the structure of the GDNF 960:984 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 2 65 dep 2v5e 392:395 arg1 code 387:390 code 387:390 code 387:390 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 8 66 contain has 1538:1540 arg2 conformation 1554:1565 a different conformation 1542:1565 a different conformation to the other three 1542:1584 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 8 66 contain has 1538:1540 arg1 persephin 1528:1536 persephin 1528:1536 persephin 1528:1536 From sequence comparison, it is suggested that neuturin (the second GFL) adopts an artemin-like conformation, while persephin has a different conformation to the other three. 19478429 1 67 theme neurotrophic 122:133 arg1 GDNF 143:146 GDNF 143:146 GDNF 143:146 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 67 theme neurotrophic 122:133 arg1 factor 135:140 Glial cell line-derived neurotrophic factor 98:140 Glial cell line-derived neurotrophic factor (GDNF) 98:147 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 68 link line-derived 109:120 arg1 GDNF 143:146 GDNF 143:146 GDNF 143:146 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 68 link line-derived 109:120 arg1 factor 135:140 Glial cell line-derived neurotrophic factor 98:140 Glial cell line-derived neurotrophic factor (GDNF) 98:147 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 2 69 from differences 413:423 arg1 complexes 467:475 GDNF-family ligand (GFL) complexes 442:475 GDNF-family ligand (GFL) complexes 442:475 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 2 69 from differences 413:423 arg1 signalling 428:437 signalling 428:437 signalling 428:437 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 7 70 theme surface 1325:1331 arg1 area 1333:1336 the buried surface area 1314:1336 the buried surface area in the monomer-monomer interface 1314:1369 Comparisons also suggest that the differences between artemin and GDNF arise from the increased curvature of the artemin ;fingers', which both increases the buried surface area in the monomer-monomer interface and changes the intermonomer bend angle. 19478429 1 71 theme GDNF-family 210:220 arg1 GFRalpha1 239:247 GFRalpha1 239:247 GFRalpha1 239:247 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 71 theme GDNF-family 210:220 arg1 alpha1 231:236 the GDNF-family receptor alpha1 206:236 the GDNF-family receptor alpha1 (GFRalpha1) 206:248 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 0 72 theme bend 68:71 arg1 angle 73:77 GFL bend angle 64:77 GFL bend angle 64:77 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 5 73 theme asymmetric 828:837 arg1 unit 839:842 the asymmetric unit 824:842 the asymmetric unit 824:842 The structures of both biological tetrameric complexes in the asymmetric unit are very similar to 2v5e and different from the artemin-GFRalpha3 structure, even though there is a small change in the structure of the GDNF. 19478429 2 74 theme previous 319:326 arg1 structure 336:344 A previous crystal structure 317:344 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) 317:396 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 1 75 theme receptor 222:229 arg1 GFRalpha1 239:247 GFRalpha1 239:247 GFRalpha1 239:247 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 1 75 theme receptor 222:229 arg1 alpha1 231:236 the GDNF-family receptor alpha1 206:236 the GDNF-family receptor alpha1 (GFRalpha1) 206:248 Glial cell line-derived neurotrophic factor (GDNF) activates the receptor tyrosine kinase RET by binding to the GDNF-family receptor alpha1 (GFRalpha1) and forming the GDNF(2)-GFRalpha1(2)-RET(2) heterohexamer complex. 19478429 0 76 theme GFL 64:66 arg1 angle 73:77 GFL bend angle 64:77 GFL bend angle 64:77 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 0 77 dep Comparison 0:9 arg1 evidence 46:53 further evidence 38:53 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 0:96 Comparison of GFL-GFRalpha complexes: further evidence relating GFL bend angle to RET signalling. 19478429 2 78 from monomers 540:547 arg1 homodimer 560:568 the GFL homodimer 552:568 the GFL homodimer 552:568 A previous crystal structure of the GDNF(2)-GFRalpha1(2) complex (PDB code 2v5e) suggested that differences in signalling in GDNF-family ligand (GFL) complexes might arise from differences in the bend angle between the two monomers in the GFL homodimer. 19478429 3 79 theme -GFRalpha1 621:630 arg1 complex 635:641 the GDNF(2)-GFRalpha1(2) complex 610:641 the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions 610:679 Here, a 2.35 A resolution structure of the GDNF(2)-GFRalpha1(2) complex crystallized with new cell dimensions is reported. 2775232 4 0 theme receptors 970:978 arg1 positions 1001:1009 the conservative positions 984:1009 the conservative positions of cysteine residues in their amino acid sequences 984:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 4 0 theme receptors 970:978 arg1 alpha-subunits 931:944 the two-chain alpha-subunits 917:944 the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors 917:978 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 5 1 theme human 1379:1383 arg1 [Poncz 1413:1418 human erythroleukaemic-cell GPIIb [Poncz 1379:1418 human erythroleukaemic-cell GPIIb [Poncz 1379:1418 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 1 2 theme single 363:368 arg1 bond 381:384 a single disulphide bond 361:384 a single disulphide bond 361:384 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 2 3 theme N-terminal 533:542 arg1 analysis 553:560 N-terminal sequence analysis 533:560 N-terminal sequence analysis of the isolated fragments 533:586 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 8 4 theme distinct 1607:1614 arg1 domains 1616:1622 distinct domains 1607:1622 distinct domains in the alpha- and beta-subunits 1607:1654 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 3 5 from residue 750:756 arg1 GPIIb 761:765 GPIIb 761:765 GPIIb 761:765 It could be established that each cysteine residue in GPIIb, beginning at alpha-Cys-56, is disulphide-bonded to its nearest neighbour in the amino acid sequence. 2775232 8 6 from beta-subunits 1642:1654 arg1 existence 1594:1602 the existence 1590:1602 the existence of distinct domains in the alpha- and beta-subunits 1590:1654 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 1 7 from surface 264:270 arg1 components 196:205 the two molecular components 178:205 the two molecular components of the inducible receptor for fibrinogen on the platelet surface 178:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 4 8 from sequences 1052:1060 arg1 positions 1001:1009 the conservative positions 984:1009 the conservative positions of cysteine residues in their amino acid sequences 984:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 4 8 from sequences 1052:1060 arg1 alpha-subunits 931:944 the two-chain alpha-subunits 917:944 the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors 917:978 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 2 9 theme some 470:473 arg1 digestion 457:465 tryptic or endoproteinase Lys-C digestion 425:465 digestion 457:465 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 2 10 theme platelet 691:698 arg1 GPIIb 700:704 platelet GPIIb 691:704 platelet GPIIb 691:704 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 8 11 from identification 1672:1685 arg1 topography 1726:1735 its external topography 1713:1735 its external topography 1713:1735 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 0 12 theme platelet 118:125 arg1 IIb 140:142 human platelet glycoprotein IIb 112:142 human platelet glycoprotein IIb 112:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 1 13 theme GPIIb 328:332 arg1 beta 334:337 GPIIb beta 328:337 GPIIb beta (22.5 kDa) 328:348 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 13 theme GPIIb 328:332 arg1 kDa 345:347 22.5 kDa 340:347 22.5 kDa 340:347 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 13 theme GPIIb 328:332 arg1 subunits 292:299 two subunits 288:299 two subunits 288:299 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 0 14 from points 81:86 arg1 alpha-subunit 95:107 the alpha-subunit 91:107 the alpha-subunit of human platelet glycoprotein IIb 91:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 4 15 theme amino 1041:1045 arg1 sequences 1052:1060 their amino acid sequences 1035:1060 their amino acid sequences 1035:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 0 16 theme IIb 140:142 arg1 alpha-subunit 95:107 the alpha-subunit 91:107 the alpha-subunit of human platelet glycoprotein IIb 91:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 1 17 theme components 196:205 arg1 one 171:173 one 171:173 one 171:173 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 17 theme components 196:205 arg1 IIb 158:160 Glycoprotein IIb 145:160 Glycoprotein IIb (GPIIb) 145:168 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 17 theme components 196:205 arg1 components 196:205 the two molecular components 178:205 the two molecular components of the inducible receptor for fibrinogen on the platelet surface 178:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 2 18 theme CNBr 491:494 arg1 peptides 496:503 the isolated CNBr peptides 478:503 the isolated CNBr peptides 478:503 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 1 19 from receptor 224:231 arg1 surface 264:270 the platelet surface 251:270 the platelet surface 251:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 0 20 gly glycoprotein 127:138 arg1 glycoprotein 127:138 human platelet glycoprotein IIb 112:142 human platelet glycoprotein IIb 112:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 2 21 theme isolated 569:576 arg1 fragments 578:586 the isolated fragments 565:586 the isolated fragments 565:586 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 4 22 from positions 1001:1009 arg1 sequences 1052:1060 their amino acid sequences 1035:1060 their amino acid sequences 1035:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 4 23 located found 1117:1121 arg2 intrachain 1067:1076 the intrachain and interchain disulphide-bond pattern 1063:1115 intrachain 1067:1076 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 4 23 located found 1117:1121 arg1 GPIIb 1131:1135 GPIIb 1131:1135 GPIIb 1131:1135 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 2 24 theme unknown 632:638 arg1 bonds 651:655 all the unknown disulphide bonds 624:655 all the unknown disulphide bonds 624:655 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 5 25 theme glycosylation 1234:1246 arg1 points 1248:1253 The N-linked glycosylation points 1221:1253 The N-linked glycosylation points found here in platelet GPIIb 1221:1282 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 25 theme glycosylation 1234:1246 arg1 same 1292:1295 same 1292:1295 same 1292:1295 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 2 26 theme GPIIb 404:408 arg1 cleavage 392:399 CNBr cleavage 387:399 CNBr cleavage 387:399 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 4 27 theme disulphide-bond 1093:1107 arg1 pattern 1109:1115 the intrachain and interchain disulphide-bond pattern 1063:1115 pattern 1109:1115 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 8 28 theme well-defined 1690:1701 arg1 points 1703:1708 well-defined points 1690:1708 well-defined points in its external topography 1690:1735 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 2 29 theme endoproteinase 436:449 arg1 digestion 457:465 tryptic or endoproteinase Lys-C digestion 425:465 digestion 457:465 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 5 30 theme GPIIb 1407:1411 arg1 [Poncz 1413:1418 human erythroleukaemic-cell GPIIb [Poncz 1379:1418 human erythroleukaemic-cell GPIIb [Poncz 1379:1418 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 2 31 theme disulphide 640:649 arg1 bonds 651:655 all the unknown disulphide bonds 624:655 all the unknown disulphide bonds 624:655 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 2 32 theme tryptic 425:431 arg1 digestion 457:465 tryptic or endoproteinase Lys-C digestion 425:465 digestion 457:465 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 2 33 theme sequence 544:551 arg1 analysis 553:560 N-terminal sequence analysis 533:560 N-terminal sequence analysis of the isolated fragments 533:586 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 8 34 theme external 1717:1724 arg1 topography 1726:1735 its external topography 1713:1735 its external topography 1713:1735 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 0 35 theme Complete 0:7 arg1 localization 9:20 Complete localization 0:20 Complete localization of the intrachain disulphide bonds 0:55 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 1 36 theme platelet 255:262 arg1 surface 264:270 the platelet surface 251:270 the platelet surface 251:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 4 37 theme adhesion 961:968 arg1 receptors 970:978 Arg-Gly-Asp adhesion receptors 949:978 Arg-Gly-Asp adhesion receptors 949:978 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 5 38 theme [Poncz 1413:1418 arg1 sequencing 1365:1374 cDNA sequencing 1360:1374 cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz 1360:1418 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 38 theme [Poncz 1413:1418 arg1 Heindenreich 1429:1440 Heindenreich 1429:1440 Heindenreich 1429:1440 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 3 39 theme nearest 823:829 arg1 neighbour 831:839 its nearest neighbour 819:839 its nearest neighbour in the amino acid sequence 819:866 It could be established that each cysteine residue in GPIIb, beginning at alpha-Cys-56, is disulphide-bonded to its nearest neighbour in the amino acid sequence. 2775232 0 40 theme intrachain 29:38 arg1 bonds 51:55 the intrachain disulphide bonds 25:55 the intrachain disulphide bonds 25:55 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 4 41 theme two-chain 921:929 arg1 alpha-subunits 931:944 the two-chain alpha-subunits 917:944 the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors 917:978 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 5 42 located found 1255:1259 arg1 GPIIb 1278:1282 platelet GPIIb 1269:1282 platelet GPIIb 1269:1282 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 42 located found 1255:1259 arg2 same 1292:1295 same 1292:1295 same 1292:1295 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 42 located found 1255:1259 arg2 points 1248:1253 The N-linked glycosylation points 1221:1253 The N-linked glycosylation points found here in platelet GPIIb 1221:1282 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 8 43 from existence 1594:1602 arg1 beta-subunits 1642:1654 beta-subunits 1642:1654 beta-subunits 1642:1654 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 8 43 from existence 1594:1602 arg1 alpha- 1631:1636 alpha- 1631:1636 alpha- 1631:1636 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 5 44 theme asparagine 1324:1333 arg1 residues 1335:1342 the five N-glycosylated asparagine residues 1300:1342 the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol 1300:1500 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 0 45 theme bonds 51:55 arg1 points 81:86 the N-glycosylation points 61:86 the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb 61:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 0 45 theme bonds 51:55 arg1 localization 9:20 Complete localization 0:20 Complete localization of the intrachain disulphide bonds 0:55 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 3 46 theme amino 848:852 arg1 sequence 859:866 the amino acid sequence 844:866 the amino acid sequence 844:866 It could be established that each cysteine residue in GPIIb, beginning at alpha-Cys-56, is disulphide-bonded to its nearest neighbour in the amino acid sequence. 2775232 8 47 from alpha- 1631:1636 arg1 existence 1594:1602 the existence 1590:1602 the existence of distinct domains in the alpha- and beta-subunits 1590:1654 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 4 48 theme residues 1023:1030 arg1 positions 1001:1009 the conservative positions 984:1009 the conservative positions of cysteine residues in their amino acid sequences 984:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 4 48 theme residues 1023:1030 arg1 alpha-subunits 931:944 the two-chain alpha-subunits 917:944 the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors 917:978 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 8 49 theme structure 1562:1570 arg1 features 1546:1553 the general features 1534:1553 the general features 1534:1553 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 1 50 theme inducible 214:222 arg1 receptor 224:231 the inducible receptor 210:231 the inducible receptor for fibrinogen on the platelet surface 210:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 4 51 theme structural 889:898 arg1 similarity 900:909 the extensive structural similarity 875:909 the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences 875:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 8 52 theme GPIIb 1575:1579 arg1 structure 1562:1570 the structure 1558:1570 the structure of GPIIb 1558:1579 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 5 53 dep Bennett 1479:1485 arg1 Biol 1497:1500 Biol 1497:1500 Biol 1497:1500 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 4 54 theme conservative 988:999 arg1 positions 1001:1009 the conservative positions 984:1009 the conservative positions of cysteine residues in their amino acid sequences 984:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 5 55 theme erythroleukaemic-cell 1385:1405 arg1 [Poncz 1413:1418 human erythroleukaemic-cell GPIIb [Poncz 1379:1418 human erythroleukaemic-cell GPIIb [Poncz 1379:1418 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 2 56 theme N-glycosylation 665:679 arg1 points 681:686 the N-glycosylation points 661:686 the N-glycosylation points in platelet GPIIb 661:704 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 8 57 theme general 1538:1544 arg1 features 1546:1553 the general features 1534:1553 the general features 1534:1553 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 1 58 theme disulphide 370:379 arg1 bond 381:384 a single disulphide bond 361:384 a single disulphide bond 361:384 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 5 59 theme cDNA 1360:1363 arg1 sequencing 1365:1374 cDNA sequencing 1360:1374 cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz 1360:1418 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 59 theme cDNA 1360:1363 arg1 Heindenreich 1429:1440 Heindenreich 1429:1440 Heindenreich 1429:1440 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 2 60 theme amino 518:522 arg1 acid 524:527 amino acid 518:527 amino acid 518:527 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 5 61 gly N-glycosylated 1309:1322 arg1 residues 1335:1342 the five N-glycosylated asparagine residues 1300:1342 the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol 1300:1500 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 0 62 theme glycoprotein 127:138 arg1 IIb 140:142 human platelet glycoprotein IIb 112:142 human platelet glycoprotein IIb 112:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 8 63 theme domains 1616:1622 arg1 existence 1594:1602 the existence 1590:1602 the existence of distinct domains in the alpha- and beta-subunits 1590:1654 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 4 64 theme acid 1047:1050 arg1 sequences 1052:1060 their amino acid sequences 1035:1060 their amino acid sequences 1035:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 1 65 theme molecular 186:194 arg1 components 196:205 the two molecular components 178:205 the two molecular components of the inducible receptor for fibrinogen on the platelet surface 178:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 2 66 theme Lys-C 451:455 arg1 digestion 457:465 tryptic or endoproteinase Lys-C digestion 425:465 digestion 457:465 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 3 67 from neighbour 831:839 arg1 sequence 859:866 the amino acid sequence 844:866 the amino acid sequence 844:866 It could be established that each cysteine residue in GPIIb, beginning at alpha-Cys-56, is disulphide-bonded to its nearest neighbour in the amino acid sequence. 2775232 1 68 from components 196:205 arg1 surface 264:270 the platelet surface 251:270 the platelet surface 251:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 4 69 from residues 1023:1030 arg1 sequences 1052:1060 their amino acid sequences 1035:1060 their amino acid sequences 1035:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 2 70 theme isolated 482:489 arg1 peptides 496:503 the isolated CNBr peptides 478:503 the isolated CNBr peptides 478:503 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 5 71 link N-linked 1225:1232 arg1 points 1248:1253 The N-linked glycosylation points 1221:1253 The N-linked glycosylation points found here in platelet GPIIb 1221:1282 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 71 link N-linked 1225:1232 arg1 same 1292:1295 same 1292:1295 same 1292:1295 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 8 72 from points 1703:1708 arg1 topography 1726:1735 its external topography 1713:1735 its external topography 1713:1735 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 4 73 theme interchain 1082:1091 arg1 pattern 1109:1115 the intrachain and interchain disulphide-bond pattern 1063:1115 pattern 1109:1115 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 5 74 theme N-linked 1225:1232 arg1 points 1248:1253 The N-linked glycosylation points 1221:1253 The N-linked glycosylation points found here in platelet GPIIb 1221:1282 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 5 74 theme N-linked 1225:1232 arg1 same 1292:1295 same 1292:1295 same 1292:1295 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 0 75 theme N-glycosylation 65:79 arg1 points 81:86 the N-glycosylation points 61:86 the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb 61:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 1 76 theme Glycoprotein 145:156 arg1 one 171:173 one 171:173 one 171:173 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 76 theme Glycoprotein 145:156 arg1 IIb 158:160 Glycoprotein IIb 145:160 Glycoprotein IIb (GPIIb) 145:168 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 76 theme Glycoprotein 145:156 arg1 components 196:205 the two molecular components 178:205 the two molecular components of the inducible receptor for fibrinogen on the platelet surface 178:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 76 theme Glycoprotein 145:156 arg1 GPIIb 163:167 GPIIb 163:167 GPIIb 163:167 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 8 77 from topography 1726:1735 arg1 identification 1672:1685 the identification 1668:1685 the identification of well-defined points in its external topography 1668:1735 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 1 78 theme GPIIb 302:306 arg1 alpha 308:312 GPIIb alpha 302:312 GPIIb alpha (114 kDa) 302:322 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 78 theme GPIIb 302:306 arg1 kDa 319:321 114 kDa 315:321 114 kDa 315:321 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 1 78 theme GPIIb 302:306 arg1 subunits 292:299 two subunits 288:299 two subunits 288:299 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 2775232 4 79 theme extensive 879:887 arg1 similarity 900:909 the extensive structural similarity 875:909 the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences 875:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 0 80 from localization 9:20 arg1 alpha-subunit 95:107 the alpha-subunit 91:107 the alpha-subunit of human platelet glycoprotein IIb 91:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 8 81 dep alpha- 1631:1636 arg1 the 1627:1629 the 1627:1629 the 1627:1629 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 3 82 theme cysteine 741:748 arg1 residue 750:756 each cysteine residue 736:756 each cysteine residue in GPIIb, beginning at alpha-Cys-56, 736:793 It could be established that each cysteine residue in GPIIb, beginning at alpha-Cys-56, is disulphide-bonded to its nearest neighbour in the amino acid sequence. 2775232 4 83 theme two-chain 1176:1184 arg1 alpha-subunits 1186:1199 all two-chain alpha-subunits 1172:1199 all two-chain alpha-subunits of these receptors 1172:1218 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 5 84 theme platelet 1269:1276 arg1 GPIIb 1278:1282 platelet GPIIb 1269:1282 platelet GPIIb 1269:1282 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 0 85 theme human 112:116 arg1 IIb 140:142 human platelet glycoprotein IIb 112:142 human platelet glycoprotein IIb 112:142 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 8 86 theme points 1703:1708 arg1 identification 1672:1685 the identification 1668:1685 the identification of well-defined points in its external topography 1668:1735 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 2 87 theme fragments 578:586 arg1 acid 524:527 amino acid 518:527 amino acid 518:527 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 2 87 theme fragments 578:586 arg1 analysis 553:560 N-terminal sequence analysis 533:560 N-terminal sequence analysis of the isolated fragments 533:586 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 4 88 theme Arg-Gly-Asp 949:959 arg1 receptors 970:978 Arg-Gly-Asp adhesion receptors 949:978 Arg-Gly-Asp adhesion receptors 949:978 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 0 89 theme disulphide 40:49 arg1 bonds 51:55 the intrachain disulphide bonds 25:55 the intrachain disulphide bonds 25:55 Complete localization of the intrachain disulphide bonds and the N-glycosylation points in the alpha-subunit of human platelet glycoprotein IIb. 2775232 4 90 from alpha-subunits 931:944 arg1 sequences 1052:1060 their amino acid sequences 1035:1060 their amino acid sequences 1035:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 2 91 theme CNBr 387:390 arg1 cleavage 392:399 CNBr cleavage 387:399 CNBr cleavage 387:399 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 2 92 from bonds 651:655 arg1 GPIIb 700:704 platelet GPIIb 691:704 platelet GPIIb 691:704 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 5 93 theme N-glycosylated 1309:1322 arg1 residues 1335:1342 the five N-glycosylated asparagine residues 1300:1342 the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol 1300:1500 The N-linked glycosylation points found here in platelet GPIIb are the same as the five N-glycosylated asparagine residues suggested after cDNA sequencing of human erythroleukaemic-cell GPIIb [Poncz, Eisman, Heindenreich, Silver, Vilaire, Surrey, Schwartz & Bennett (1987) J. Biol. 2775232 2 94 dep together 411:418 arg1 with 420:423 with 420:423 with 420:423 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 3 95 theme acid 854:857 arg1 sequence 859:866 the amino acid sequence 844:866 the amino acid sequence 844:866 It could be established that each cysteine residue in GPIIb, beginning at alpha-Cys-56, is disulphide-bonded to its nearest neighbour in the amino acid sequence. 2775232 4 96 theme receptors 1210:1218 arg1 alpha-subunits 1186:1199 all two-chain alpha-subunits 1172:1199 all two-chain alpha-subunits of these receptors 1172:1218 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 2 97 from points 681:686 arg1 GPIIb 700:704 platelet GPIIb 691:704 platelet GPIIb 691:704 CNBr cleavage of GPIIb, together with tryptic or endoproteinase Lys-C digestion of some of the isolated CNBr peptides, followed by amino acid and N-terminal sequence analysis of the isolated fragments, allowed us to locate unambiguously all the unknown disulphide bonds and the N-glycosylation points in platelet GPIIb. 2775232 4 98 theme cysteine 1014:1021 arg1 residues 1023:1030 cysteine residues 1014:1030 cysteine residues in their amino acid sequences 1014:1060 Given the extensive structural similarity among the two-chain alpha-subunits of Arg-Gly-Asp adhesion receptors and the conservative positions of cysteine residues in their amino acid sequences, the intrachain and interchain disulphide-bond pattern found here in GPIIb will most probably be conserved in all two-chain alpha-subunits of these receptors. 2775232 8 99 from domains 1616:1622 arg1 beta-subunits 1642:1654 beta-subunits 1642:1654 beta-subunits 1642:1654 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 8 99 from domains 1616:1622 arg1 alpha- 1631:1636 alpha- 1631:1636 alpha- 1631:1636 Some of the general features of the structure of GPIIb, such as the existence of distinct domains in the alpha- and beta-subunits, as well as the identification of well-defined points in its external topography, are discussed. 2775232 1 100 theme receptor 224:231 arg1 components 196:205 the two molecular components 178:205 the two molecular components of the inducible receptor for fibrinogen on the platelet surface 178:270 Glycoprotein IIb (GPIIb), one of the two molecular components of the inducible receptor for fibrinogen on the platelet surface, is formed from two subunits, GPIIb alpha (114 kDa) and GPIIb beta (22.5 kDa), joined by a single disulphide bond. 1472036 0 0 theme domain 88:93 arg1 alpha-subunits 52:65 the alpha-subunits 48:65 the alpha-subunits of the extracellular domain of the insulin receptor 48:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 2 1 theme soluble 331:337 arg1 receptor 375:382 the insulin receptor 363:382 the insulin receptor 363:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 2 1 theme soluble 331:337 arg1 domain 353:358 the soluble extracellular domain 327:358 the soluble extracellular domain of the insulin receptor 327:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 0 2 theme extracellular 74:86 arg1 domain 88:93 the extracellular domain 70:93 the extracellular domain of the insulin receptor 70:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 0 2 theme extracellular 74:86 arg1 receptor 110:117 the insulin receptor 98:117 the insulin receptor 98:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 3 3 contain containing 505:514 arg2 Cys435 516:521 Cys435 516:521 Cys435 516:521 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 3 3 contain containing 505:514 arg1 peptides 496:503 The peptides 492:503 The peptides containing Cys435 and Cys468 492:532 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 3 3 contain containing 505:514 arg2 Cys468 527:532 Cys468 527:532 Cys468 527:532 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 4 4 theme other 804:808 arg1 sequences 810:818 no other sequences 801:818 no other sequences containing cysteine 801:838 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 1 5 contain containing 272:281 arg2 Cys524 302:307 Cys524 302:307 Cys524 302:307 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 1 5 contain containing 272:281 arg2 Cys468 291:296 Cys468 291:296 Cys468 291:296 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 1 5 contain containing 272:281 arg2 Cys435 283:288 Cys435 283:288 Cys435 283:288 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 1 5 contain containing 272:281 arg1 region 265:270 the region 261:270 the region containing Cys435, Cys468 and Cys524 261:307 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 4 6 contain has 772:774 arg2 thiols 784:789 no free thiols 776:789 no free thiols 776:789 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 6 contain has 772:774 arg1 domain 741:746 the extracellular domain 723:746 the extracellular domain of the insulin receptor 723:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 6 contain has 772:774 arg1 receptor 763:770 the insulin receptor 751:770 the insulin receptor 751:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 0 7 theme receptor 110:117 arg1 domain 88:93 the extracellular domain 70:93 the extracellular domain of the insulin receptor 70:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 0 7 theme receptor 110:117 arg1 receptor 110:117 the insulin receptor 98:117 the insulin receptor 98:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 4 8 theme extracellular 727:739 arg1 domain 741:746 the extracellular domain 723:746 the extracellular domain of the insulin receptor 723:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 8 theme extracellular 727:739 arg1 receptor 763:770 the insulin receptor 751:770 the insulin receptor 751:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 2 9 theme receptor 375:382 arg1 receptor 375:382 the insulin receptor 363:382 the insulin receptor 363:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 2 9 theme receptor 375:382 arg1 domain 353:358 the soluble extracellular domain 327:358 the soluble extracellular domain of the insulin receptor 327:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 0 10 theme insulin 102:108 arg1 receptor 110:117 the insulin receptor 98:117 the insulin receptor 98:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 3 11 theme peptide 667:673 arg1 sequence 651:658 the sequence 647:658 the sequence of the peptide containing Cys524 647:691 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 1 12 theme insulin 156:162 arg1 receptor 164:171 the insulin receptor 152:171 the insulin receptor 152:171 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 2 13 theme resulting 435:443 arg1 peptides 445:452 the resulting peptides 431:452 the resulting peptides 431:452 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 3 14 theme same 550:553 arg1 fraction 555:562 the same fraction 546:562 the same fraction 546:562 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 1 15 theme receptor 164:171 arg1 structure 139:147 The alpha 2 beta 2 structure 120:147 The alpha 2 beta 2 structure of the insulin receptor 120:171 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 0 16 theme disulfide 20:28 arg1 bridge 30:35 a disulfide bridge 18:35 a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor 18:117 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 1 17 from alpha-subunits 243:256 arg1 region 265:270 the region 261:270 the region containing Cys435, Cys468 and Cys524 261:307 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 4 18 located found 845:849 arg2 sequences 810:818 no other sequences 801:818 no other sequences containing cysteine 801:838 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 18 located found 845:849 arg1 fractions 860:868 these fractions 854:868 these fractions 854:868 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 2 19 theme succinylated 389:400 arg1 trypsin 402:408 succinylated trypsin 389:408 succinylated trypsin 389:408 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 0 20 theme bridge 30:35 arg1 Identification 0:13 Identification 0:13 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 0:118 Identification of a disulfide bridge connecting the alpha-subunits of the extracellular domain of the insulin receptor. 1472036 4 21 theme receptor 763:770 arg1 domain 741:746 the extracellular domain 723:746 the extracellular domain of the insulin receptor 723:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 21 theme receptor 763:770 arg1 receptor 763:770 the insulin receptor 751:770 the insulin receptor 751:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 22 theme other 939:943 arg1 alpha-subunit 945:957 the other alpha-subunit 935:957 the other alpha-subunit 935:957 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 23 theme free 779:782 arg1 thiols 784:789 no free thiols 776:789 no free thiols 776:789 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 3 24 contain containing 675:684 arg2 Cys524 686:691 Cys524 686:691 Cys524 686:691 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 3 24 contain containing 675:684 arg1 peptide 667:673 the peptide 663:673 the peptide containing Cys524 663:691 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 4 25 from Cys524 925:930 arg1 alpha-subunit 945:957 the other alpha-subunit 935:957 the other alpha-subunit 935:957 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 1 26 theme alpha 124:128 arg1 beta 132:135 alpha 2 beta 2 124:137 The alpha 2 beta 2 structure of the insulin receptor 120:171 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 3 27 theme disulfide 598:606 arg1 bond 608:611 a disulfide bond 596:611 a disulfide bond 596:611 The peptides containing Cys435 and Cys468 appeared in the same fraction, indicating that these two form a disulfide bond, and in another fraction we found the sequence of the peptide containing Cys524. 1472036 2 28 theme fractions 481:489 arg1 number 471:476 a number 469:476 a number of fractions 469:489 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 4 29 theme insulin 755:761 arg1 receptor 763:770 the insulin receptor 751:770 the insulin receptor 751:770 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 1 30 theme beta 132:135 arg1 structure 139:147 The alpha 2 beta 2 structure 120:147 The alpha 2 beta 2 structure of the insulin receptor 120:171 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 2 31 theme insulin 367:373 arg1 receptor 375:382 the insulin receptor 363:382 the insulin receptor 363:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 1 32 theme disulfide 214:222 arg1 bridge 224:229 one disulfide bridge 210:229 one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524 210:307 The alpha 2 beta 2 structure of the insulin receptor has previously been shown to involve one disulfide bridge between the alpha-subunits in the region containing Cys435, Cys468 and Cys524. 1472036 4 33 theme disulfide 903:911 arg1 bond 913:916 a disulfide bond 901:916 a disulfide bond to the Cys524 in the other alpha-subunit 901:957 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 2 34 theme extracellular 339:351 arg1 receptor 375:382 the insulin receptor 363:382 the insulin receptor 363:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 2 34 theme extracellular 339:351 arg1 domain 353:358 the soluble extracellular domain 327:358 the soluble extracellular domain of the insulin receptor 327:382 We have digested the soluble extracellular domain of the insulin receptor with succinylated trypsin, partially separated the resulting peptides, and sequenced a number of fractions. 1472036 4 35 contain containing 820:829 arg2 cysteine 831:838 cysteine 831:838 cysteine 831:838 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 1472036 4 35 contain containing 820:829 arg1 sequences 810:818 no other sequences 801:818 no other sequences containing cysteine 801:838 Since it has been shown that the extracellular domain of the insulin receptor has no free thiols and since no other sequences containing cysteine were found in these fractions, we conclude that Cys524 forms a disulfide bond to the Cys524 in the other alpha-subunit. 12122212 3 0 theme fragment 425:432 arg1 structure 393:401 the crystal structure 381:401 the crystal structure of the CD21 SCR1-SCR2 fragment 381:432 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 1 1 theme tumor 193:197 arg1 virus 199:203 a human tumor virus 185:203 a human tumor virus 185:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 1 theme tumor 193:197 arg1 virus 172:176 Epstein-Barr virus 159:176 Epstein-Barr virus (EBV) 159:182 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 7 2 theme additional 1235:1244 arg1 epitopes 1246:1253 additional epitopes 1235:1253 additional epitopes for C3d binding 1235:1269 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 7 3 theme protein 1124:1130 arg1 deglycosylation 1101:1115 deglycosylation 1101:1115 deglycosylation of the protein 1101:1130 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 5 4 theme compact 845:851 arg1 conformations 862:874 compact V-shaped conformations 845:874 compact V-shaped conformations 845:874 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 7 5 theme extended 1199:1206 arg1 conformation 1208:1219 a more extended conformation 1192:1219 a more extended conformation 1192:1219 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 2 6 theme consensus 231:239 arg1 SCR1-SCR2 250:258 SCR1-SCR2 250:258 SCR1-SCR2 250:258 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 2 6 theme consensus 231:239 arg1 repeats 241:247 The N-terminal two short consensus repeats 206:247 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor 206:275 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 0 7 theme C3d 77:79 arg1 binding 81:87 C3d binding 77:87 C3d binding 77:87 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 12122212 2 8 theme short 225:229 arg1 SCR1-SCR2 250:258 SCR1-SCR2 250:258 SCR1-SCR2 250:258 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 2 8 theme short 225:229 arg1 repeats 241:247 The N-terminal two short consensus repeats 206:247 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor 206:275 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 7 9 theme physiologic 1142:1152 arg1 glycosylation 1154:1166 physiologic glycosylation 1142:1166 physiologic glycosylation of CD21 1142:1174 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 3 10 theme CD21 410:413 arg1 fragment 425:432 the CD21 SCR1-SCR2 fragment 406:432 the CD21 SCR1-SCR2 fragment 406:432 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 3 11 attach present 373:379 arg2 we 370:371 we 370:371 we 370:371 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 3 11 attach present 373:379 arg1 absence 441:447 the absence 437:447 the absence of ligand 437:457 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 4 12 theme CD21 559:562 arg1 analysis 526:533 a functional analysis 513:533 a functional analysis of wild-type and mutant CD21 and molecular modeling 513:585 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 7 13 theme V-shaped 1065:1072 arg1 conformation 1074:1085 the V-shaped conformation 1061:1085 the V-shaped conformation 1061:1085 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 2 14 theme N-terminal 210:219 arg1 SCR1-SCR2 250:258 SCR1-SCR2 250:258 SCR1-SCR2 250:258 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 2 14 theme N-terminal 210:219 arg1 repeats 241:247 The N-terminal two short consensus repeats 206:247 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor 206:275 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 1 15 theme cellular 137:144 arg1 receptor 146:153 the cellular receptor 133:153 the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus 133:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 15 theme cellular 137:144 arg1 type 116:119 Human complement receptor type 2 90:121 Human complement receptor type 2 (CD21) 90:128 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 0 16 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 0:88 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 12122212 6 17 theme SCR1-SCR2 949:957 arg1 interface 959:967 the SCR1-SCR2 interface 945:967 the SCR1-SCR2 interface 945:967 However, our analysis reveals a surprising degree of flexibility at the SCR1-SCR2 interface, suggesting interactions between the two domains are not specific. 12122212 4 18 theme molecular 568:576 arg1 modeling 578:585 molecular modeling 568:585 molecular modeling 568:585 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 4 19 theme EBV 620:622 arg1 attachment 624:633 EBV attachment 620:633 EBV attachment 620:633 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 7 20 theme CD21 1171:1174 arg1 glycosylation 1154:1166 physiologic glycosylation 1142:1166 physiologic glycosylation of CD21 1142:1174 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 6 21 theme flexibility 930:940 arg1 degree 920:925 a surprising degree 907:925 a surprising degree of flexibility 907:940 However, our analysis reveals a surprising degree of flexibility at the SCR1-SCR2 interface, suggesting interactions between the two domains are not specific. 12122212 2 22 theme ligand 353:358 arg1 C3d 360:362 the natural CD21 ligand C3d 336:362 the natural CD21 ligand C3d 336:362 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 3 23 theme crystal 385:391 arg1 structure 393:401 the crystal structure 381:401 the crystal structure of the CD21 SCR1-SCR2 fragment 381:432 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 2 24 gly glycoprotein 299:310 arg1 glycoprotein 299:310 the EBV glycoprotein gp350/220 291:320 the EBV glycoprotein gp350/220 291:320 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 1 25 theme Human 90:94 arg1 type 116:119 Human complement receptor type 2 90:121 Human complement receptor type 2 (CD21) 90:128 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 25 theme Human 90:94 arg1 CD21 124:127 CD21 124:127 CD21 124:127 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 25 theme Human 90:94 arg1 receptor 146:153 the cellular receptor 133:153 the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus 133:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 2 26 theme CD21 348:351 arg1 C3d 360:362 the natural CD21 ligand C3d 336:362 the natural CD21 ligand C3d 336:362 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 0 27 theme CD21 31:34 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 0:88 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 12122212 2 28 theme receptor 268:275 arg1 SCR1-SCR2 250:258 SCR1-SCR2 250:258 SCR1-SCR2 250:258 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 2 28 theme receptor 268:275 arg1 repeats 241:247 The N-terminal two short consensus repeats 206:247 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor 206:275 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 6 29 theme surprising 909:918 arg1 degree 920:925 a surprising degree 907:925 a surprising degree of flexibility 907:940 However, our analysis reveals a surprising degree of flexibility at the SCR1-SCR2 interface, suggesting interactions between the two domains are not specific. 12122212 7 30 theme C3d 1259:1261 arg1 binding 1263:1269 C3d binding 1259:1269 C3d binding 1259:1269 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 2 31 theme natural 340:346 arg1 C3d 360:362 the natural CD21 ligand C3d 336:362 the natural CD21 ligand C3d 336:362 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 0 32 theme human 25:29 arg1 CD21 31:34 human CD21 25:34 human CD21 25:34 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 12122212 5 33 theme determined 746:755 arg1 structure 757:765 the previously determined structure 731:765 the previously determined structure of CD21 SCR1-SCR2 in complex 731:794 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 5 34 with comparison 715:724 arg1 C3d 801:803 C3d 801:803 C3d 801:803 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 5 34 with comparison 715:724 arg1 structure 757:765 the previously determined structure 731:765 the previously determined structure of CD21 SCR1-SCR2 in complex 731:794 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 4 35 theme wild-type 538:546 arg1 CD21 559:562 wild-type and mutant CD21 538:562 wild-type and mutant CD21 538:562 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 5 36 theme CD21 770:773 arg1 SCR1-SCR2 775:783 CD21 SCR1-SCR2 770:783 CD21 SCR1-SCR2 770:783 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 1 37 theme complement 96:105 arg1 type 116:119 Human complement receptor type 2 90:121 Human complement receptor type 2 (CD21) 90:128 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 37 theme complement 96:105 arg1 CD21 124:127 CD21 124:127 CD21 124:127 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 37 theme complement 96:105 arg1 receptor 146:153 the cellular receptor 133:153 the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus 133:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 7 38 gly glycosylation 1154:1166 arg1 CD21 1171:1174 CD21 1171:1174 CD21 1171:1174 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 1 39 theme receptor 107:114 arg1 type 116:119 Human complement receptor type 2 90:121 Human complement receptor type 2 (CD21) 90:128 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 39 theme receptor 107:114 arg1 CD21 124:127 CD21 124:127 CD21 124:127 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 39 theme receptor 107:114 arg1 receptor 146:153 the cellular receptor 133:153 the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus 133:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 4 40 theme likely 602:607 arg1 region 609:614 a likely region 600:614 a likely region for EBV attachment 600:633 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 5 41 theme V-shaped 853:860 arg1 conformations 862:874 compact V-shaped conformations 845:874 compact V-shaped conformations 845:874 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 3 42 theme ligand 452:457 arg1 absence 441:447 the absence 437:447 the absence of ligand 437:457 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 0 43 dep structure 12:20 arg1 Implications 37:48 Implications 37:48 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 0:88 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 12122212 5 44 from structure 757:765 arg1 complex 788:794 complex 788:794 complex 788:794 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 4 45 theme modeling 578:585 arg1 analysis 526:533 a functional analysis 513:533 a functional analysis of wild-type and mutant CD21 and molecular modeling 513:585 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 0 46 theme Epstein-Barr 54:65 arg1 virus 67:71 Epstein-Barr virus 54:71 Epstein-Barr virus 54:71 The crystal structure of human CD21: Implications for Epstein-Barr virus and C3d binding. 12122212 5 47 theme SCR1-SCR2 775:783 arg1 structure 757:765 the previously determined structure 731:765 the previously determined structure of CD21 SCR1-SCR2 in complex 731:794 A comparison with the previously determined structure of CD21 SCR1-SCR2 in complex with C3d shows that, in both cases, CD21 assumes compact V-shaped conformations. 12122212 1 48 theme Epstein-Barr 159:170 arg1 virus 172:176 Epstein-Barr virus 159:176 Epstein-Barr virus (EBV) 159:182 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 48 theme Epstein-Barr 159:170 arg1 virus 199:203 a human tumor virus 185:203 a human tumor virus 185:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 48 theme Epstein-Barr 159:170 arg1 EBV 179:181 EBV 179:181 EBV 179:181 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 2 49 theme glycoprotein 299:310 arg1 gp350/220 312:320 the EBV glycoprotein gp350/220 291:320 the EBV glycoprotein gp350/220 291:320 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 4 50 theme mutant 552:557 arg1 CD21 559:562 wild-type and mutant CD21 538:562 wild-type and mutant CD21 538:562 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 4 51 theme functional 515:524 arg1 analysis 526:533 a functional analysis 513:533 a functional analysis of wild-type and mutant CD21 and molecular modeling 513:585 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 7 52 gly deglycosylation 1101:1115 arg1 protein 1124:1130 the protein 1120:1130 the protein 1120:1130 We present evidence that the V-shaped conformation is induced by deglycosylation of the protein, and that physiologic glycosylation of CD21 would result in a more extended conformation, perhaps with additional epitopes for C3d binding. 12122212 4 53 with interaction 691:701 arg1 C3d 708:710 C3d 708:710 C3d 708:710 Based on a functional analysis of wild-type and mutant CD21 and molecular modeling, we identify a likely region for EBV attachment and demonstrate that this region is not involved in the interaction with C3d. 12122212 2 54 theme EBV 295:297 arg1 gp350/220 312:320 the EBV glycoprotein gp350/220 291:320 the EBV glycoprotein gp350/220 291:320 The N-terminal two short consensus repeats (SCR1-SCR2) of the receptor interact with the EBV glycoprotein gp350/220 and also with the natural CD21 ligand C3d. 12122212 3 55 theme SCR1-SCR2 415:423 arg1 fragment 425:432 the CD21 SCR1-SCR2 fragment 406:432 the CD21 SCR1-SCR2 fragment 406:432 Here we present the crystal structure of the CD21 SCR1-SCR2 fragment in the absence of ligand and demonstrate that it is able to bind EBV. 12122212 1 56 theme human 187:191 arg1 virus 199:203 a human tumor virus 185:203 a human tumor virus 185:203 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 12122212 1 56 theme human 187:191 arg1 virus 172:176 Epstein-Barr virus 159:176 Epstein-Barr virus (EBV) 159:182 Human complement receptor type 2 (CD21) is the cellular receptor for Epstein-Barr virus (EBV), a human tumor virus. 1709161 11 0 theme gastrointestinal 1390:1405 arg1 antigen 1424:1430 a gastrointestinal tumor-associated antigen 1388:1430 a gastrointestinal tumor-associated antigen 1388:1430 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 2 1 theme NH2-terminal 303:314 arg1 identical 362:370 identical 362:370 identical 362:370 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 2 1 theme NH2-terminal 303:314 arg1 acids 322:326 The 15 NH2-terminal amino acids 296:326 The 15 NH2-terminal amino acids of the 24-kDa binding protein 296:356 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 8 2 theme amino 994:998 arg1 acids 1000:1004 the 213-289 amino acids 982:1004 the 213-289 amino acids 982:1004 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 6 3 theme amino 787:791 arg1 acids 793:797 213 and 237 amino acids 775:797 acids 793:797 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 12 4 theme IGFBPs 1525:1530 arg1 cDNAs 1505:1509 The cDNAs 1501:1509 The cDNAs of five human IGFBPs 1501:1530 The cDNAs of five human IGFBPs are now available. 1709161 12 4 theme IGFBPs 1525:1530 arg1 available 1540:1548 available 1540:1548 available 1540:1548 The cDNAs of five human IGFBPs are now available. 1709161 6 5 dep respectively 800:811 arg1 Mr 814:815 Mr 814:815 Mr = 22,610 and 25,980 814:835 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 4 6 attach derived 596:602 arg2 sequences 586:594 The complete amino acid sequences 562:594 The complete amino acid sequences derived from the cloned cDNAs 562:624 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 4 6 attach derived 596:602 arg1 cDNAs 620:624 the cloned cDNAs 609:624 the cloned cDNAs 609:624 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 1 7 theme kDa 242:244 arg1 nonreduced 247:256 nonreduced 247:256 nonreduced 247:256 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 7 theme kDa 242:244 arg1 masses 214:219 apparent molecular masses 195:219 apparent molecular masses of 24, 28-30, and 30 kDa 195:244 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 4 8 theme amino 575:579 arg1 sequences 586:594 The complete amino acid sequences 562:594 The complete amino acid sequences derived from the cloned cDNAs 562:624 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 11 9 theme IGFBPs 1334:1339 arg1 domains 1356:1362 10 repetitive domains 1342:1362 10 repetitive domains of human thyroglobulin 1342:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 9 theme IGFBPs 1334:1339 arg1 antigen 1492:1498 class II histocompatibility antigen 1464:1498 the class II histocompatibility antigen 1460:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 9 theme IGFBPs 1334:1339 arg1 antigen 1424:1430 a gastrointestinal tumor-associated antigen 1388:1430 a gastrointestinal tumor-associated antigen 1388:1430 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 9 theme IGFBPs 1334:1339 arg1 IGFBPs 1334:1339 the five IGFBPs 1325:1339 the five IGFBPs 1325:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 9 theme IGFBPs 1334:1339 arg1 thyroglobulin 1373:1385 human thyroglobulin 1367:1385 human thyroglobulin 1367:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 9 theme IGFBPs 1334:1339 arg1 quarter 1314:1320 the COOH-terminal quarter 1296:1320 the COOH-terminal quarter of the five IGFBPs 1296:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 9 theme IGFBPs 1334:1339 arg1 chain 1451:1455 the invariant chain 1437:1455 the invariant chain of the class II histocompatibility antigen 1437:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 3 10 theme 30-kDa 500:505 arg1 BP 507:508 the 30-kDa BP 496:508 the 30-kDa BP 496:508 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 10 theme 30-kDa 500:505 arg1 form 530:533 the glycosylated form 513:533 the glycosylated form of the isolated 24-kDa BP 513:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 11 gly glycosylated 517:528 arg1 BP 507:508 the 30-kDa BP 496:508 the 30-kDa BP 496:508 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 11 gly glycosylated 517:528 arg1 BP 558:559 the isolated 24-kDa BP 538:559 the isolated 24-kDa BP 538:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 11 gly glycosylated 517:528 arg1 form 530:533 the glycosylated form 513:533 the glycosylated form of the isolated 24-kDa BP 513:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 0 12 theme human 119:123 arg1 serum 125:129 adult human serum 113:129 adult human serum 113:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 1 13 theme apparent 195:202 arg1 nonreduced 247:256 nonreduced 247:256 nonreduced 247:256 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 13 theme apparent 195:202 arg1 masses 214:219 apparent molecular masses 195:219 apparent molecular masses of 24, 28-30, and 30 kDa 195:244 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 8 14 theme cysteines 962:970 arg1 16 946:947 16 946:947 16 946:947 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 8 14 theme cysteines 962:970 arg1 % 1016:1016 12.8-17.1% 1007:1016 12.8-17.1% 1007:1016 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 8 14 theme cysteines 962:970 arg1 cysteines 962:970 the 16-20 cysteines 952:970 the 16-20 cysteines 952:970 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 3 15 theme glycosylated 517:528 arg1 BP 507:508 the 30-kDa BP 496:508 the 30-kDa BP 496:508 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 15 theme glycosylated 517:528 arg1 form 530:533 the glycosylated form 513:533 the glycosylated form of the isolated 24-kDa BP 513:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 7 16 theme BP-4 876:879 arg1 NH2- 843:846 NH2- 843:846 NH2- 843:846 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 7 16 theme BP-4 876:879 arg1 thirds 866:871 COOH-terminal thirds 852:871 COOH-terminal thirds 852:871 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 6 17 theme = 817:817 arg1 Mr 814:815 Mr 814:815 Mr = 22,610 and 25,980 814:835 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 18 dep = 817:817 arg1 25,980 830:835 25,980 830:835 25,980 830:835 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 18 dep = 817:817 arg1 22,610 819:824 22,610 819:824 22,610 819:824 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 9 19 theme amino 1060:1064 arg1 -5 1148:1149 -5 1148:1149 -5 1148:1149 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 19 theme amino 1060:1064 arg1 -3 1140:1141 -3 1140:1141 -3 1140:1141 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 19 theme amino 1060:1064 arg1 positions 1071:1079 10 amino acid positions 1057:1079 10 amino acid positions located in the NH2-terminal region and shared by BP-1 1057:1133 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 19 theme amino 1060:1064 arg1 -2 1136:1137 -2 1136:1137 -2 1136:1137 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 13 20 from production 1588:1597 arg1 quantities 1613:1622 sufficient quantities 1602:1622 sufficient quantities 1602:1622 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 4 21 theme new 640:642 arg1 IGFBPs 644:649 two new IGFBPs 636:649 two new IGFBPs 636:649 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 11 22 theme invariant 1441:1449 arg1 chain 1451:1455 the invariant chain 1437:1455 the invariant chain of the class II histocompatibility antigen 1437:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 3 23 theme isolated 542:549 arg1 BP 558:559 the isolated 24-kDa BP 538:559 the isolated 24-kDa BP 538:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 13 24 theme sufficient 1602:1611 arg1 quantities 1613:1622 sufficient quantities 1602:1622 sufficient quantities 1602:1622 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 0 25 theme insulin-like 55:66 arg1 factor 75:80 30-kDa insulin-like growth factor 48:80 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 1 26 theme growth 151:156 arg1 IGFBP 183:187 IGFBP 183:187 IGFBP 183:187 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 26 theme growth 151:156 arg1 proteins 173:180 Three insulin-like growth factor binding proteins 132:180 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, 132:257 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 4 27 theme cloned 613:618 arg1 cDNAs 620:624 the cloned cDNAs 609:624 the cloned cDNAs 609:624 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 2 28 with identical 362:370 arg1 those 377:381 those 377:381 those 377:381 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 0 29 theme factor 75:80 arg1 proteins 90:97 two new 30-kDa insulin-like growth factor binding proteins 40:97 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 9 30 theme NH2-terminal 1096:1107 arg1 region 1109:1114 the NH2-terminal region 1092:1114 the NH2-terminal region 1092:1114 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 1 31 theme binding 165:171 arg1 IGFBP 183:187 IGFBP 183:187 IGFBP 183:187 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 31 theme binding 165:171 arg1 proteins 173:180 Three insulin-like growth factor binding proteins 132:180 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, 132:257 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 0 32 theme proteins 90:97 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 0 32 theme proteins 90:97 arg1 cloning 29:35 molecular cloning 19:35 molecular cloning 19:35 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 2 33 theme protein 350:356 arg1 identical 362:370 identical 362:370 identical 362:370 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 2 33 theme protein 350:356 arg1 acids 322:326 The 15 NH2-terminal amino acids 296:326 The 15 NH2-terminal amino acids of the 24-kDa binding protein 296:356 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 10 34 theme BP-4 1237:1240 arg1 affinity 1225:1232 the preferential affinity 1208:1232 the preferential affinity of BP-4 for IGF II 1208:1251 These differences may account for the preferential affinity of BP-4 for IGF II. 1709161 2 35 theme 24-kDa 335:340 arg1 protein 350:356 the 24-kDa binding protein 331:356 the 24-kDa binding protein 331:356 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 13 36 from expression 1573:1582 arg1 quantities 1613:1622 sufficient quantities 1602:1622 sufficient quantities 1602:1622 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 7 37 dep NH2- 843:846 arg1 The 839:841 The 839:841 The 839:841 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 3 38 theme apparent 405:412 arg1 mass 424:427 The apparent molecular mass 401:427 The apparent molecular mass of the latter 401:441 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 0 39 theme molecular 19:27 arg1 cloning 29:35 molecular cloning 19:35 molecular cloning 19:35 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 6 40 theme mature 759:764 arg1 proteins 766:773 the mature proteins 755:773 the mature proteins 755:773 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 40 theme mature 759:764 arg1 21 751:752 21 751:752 21 751:752 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 41 dep 27 744:745 arg1 acids 793:797 213 and 237 amino acids 775:797 acids 793:797 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 3 42 theme BP 558:559 arg1 BP 507:508 the 30-kDa BP 496:508 the 30-kDa BP 496:508 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 42 theme BP 558:559 arg1 form 530:533 the glycosylated form 513:533 the glycosylated form of the isolated 24-kDa BP 513:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 0 43 theme new 44:46 arg1 proteins 90:97 two new 30-kDa insulin-like growth factor binding proteins 40:97 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 13 44 dep in 1628:1629 arg1 vivo 1631:1634 vivo 1631:1634 vivo 1631:1634 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 6 45 theme prepeptide 700:709 arg1 sequences 711:719 The prepeptide sequences 696:719 The prepeptide sequences of BP-4 and -5 696:734 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 11 46 theme repetitive 1345:1354 arg1 domains 1356:1362 10 repetitive domains 1342:1362 10 repetitive domains of human thyroglobulin 1342:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 46 theme repetitive 1345:1354 arg1 thyroglobulin 1373:1385 human thyroglobulin 1367:1385 human thyroglobulin 1367:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 46 theme repetitive 1345:1354 arg1 IGFBPs 1334:1339 the five IGFBPs 1325:1339 the five IGFBPs 1325:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 46 theme repetitive 1345:1354 arg1 antigen 1492:1498 class II histocompatibility antigen 1464:1498 the class II histocompatibility antigen 1460:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 domains 1356:1362 10 repetitive domains 1342:1362 10 repetitive domains of human thyroglobulin 1342:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 antigen 1492:1498 class II histocompatibility antigen 1464:1498 the class II histocompatibility antigen 1460:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 antigen 1424:1430 a gastrointestinal tumor-associated antigen 1388:1430 a gastrointestinal tumor-associated antigen 1388:1430 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 IGFBPs 1334:1339 the five IGFBPs 1325:1339 the five IGFBPs 1325:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 thyroglobulin 1373:1385 human thyroglobulin 1367:1385 human thyroglobulin 1367:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 quarter 1314:1320 the COOH-terminal quarter 1296:1320 the COOH-terminal quarter of the five IGFBPs 1296:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 47 theme antigen 1492:1498 arg1 chain 1451:1455 the invariant chain 1437:1455 the invariant chain of the class II histocompatibility antigen 1437:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 4 48 theme complete 566:573 arg1 sequences 586:594 The complete amino acid sequences 562:594 The complete amino acid sequences derived from the cloned cDNAs 562:624 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 9 49 from located 1081:1087 arg1 region 1109:1114 the NH2-terminal region 1092:1114 the NH2-terminal region 1092:1114 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 domains 1356:1362 10 repetitive domains 1342:1362 10 repetitive domains of human thyroglobulin 1342:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 antigen 1492:1498 class II histocompatibility antigen 1464:1498 the class II histocompatibility antigen 1460:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 antigen 1424:1430 a gastrointestinal tumor-associated antigen 1388:1430 a gastrointestinal tumor-associated antigen 1388:1430 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 IGFBPs 1334:1339 the five IGFBPs 1325:1339 the five IGFBPs 1325:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 thyroglobulin 1373:1385 human thyroglobulin 1367:1385 human thyroglobulin 1367:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 quarter 1314:1320 the COOH-terminal quarter 1296:1320 the COOH-terminal quarter of the five IGFBPs 1296:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 11 50 theme thyroglobulin 1373:1385 arg1 chain 1451:1455 the invariant chain 1437:1455 the invariant chain of the class II histocompatibility antigen 1437:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 2 51 theme amino 316:320 arg1 identical 362:370 identical 362:370 identical 362:370 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 2 51 theme amino 316:320 arg1 acids 322:326 The 15 NH2-terminal amino acids 296:326 The 15 NH2-terminal amino acids of the 24-kDa binding protein 296:356 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 11 52 theme tumor-associated 1407:1422 arg1 antigen 1424:1430 a gastrointestinal tumor-associated antigen 1388:1430 a gastrointestinal tumor-associated antigen 1388:1430 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 6 53 theme BP-4 724:727 arg1 sequences 711:719 The prepeptide sequences 696:719 The prepeptide sequences of BP-4 and -5 696:734 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 54 theme 237 783:785 arg1 acids 793:797 213 and 237 amino acids 775:797 acids 793:797 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 11 55 theme COOH-terminal 1300:1312 arg1 quarter 1314:1320 the COOH-terminal quarter 1296:1320 the COOH-terminal quarter of the five IGFBPs 1296:1339 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 8 56 theme acids 1000:1004 arg1 acids 1000:1004 the 213-289 amino acids 982:1004 the 213-289 amino acids 982:1004 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 8 56 theme acids 1000:1004 arg1 37 976:977 37 976:977 37 976:977 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 6 57 theme 213 775:777 arg1 acids 793:797 213 and 237 amino acids 775:797 acids 793:797 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 8 58 theme mature 1045:1050 arg1 BPs 1052:1054 all five mature BPs 1036:1054 all five mature BPs 1036:1054 16 of the 16-20 cysteines and 37 of the 213-289 amino acids (12.8-17.1%) are conserved in all five mature BPs. 1709161 7 59 theme human 935:939 arg1 BPs 941:943 the other three human BPs 919:943 the other three human BPs 919:943 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 1 60 attach isolated 269:276 arg2 proteins 173:180 Three insulin-like growth factor binding proteins 132:180 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, 132:257 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 60 attach isolated 269:276 arg1 serum 289:293 human serum 283:293 human serum 283:293 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 60 attach isolated 269:276 arg2 IGFBP 183:187 IGFBP 183:187 IGFBP 183:187 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 12 61 theme human 1519:1523 arg1 IGFBPs 1525:1530 five human IGFBPs 1514:1530 five human IGFBPs 1514:1530 The cDNAs of five human IGFBPs are now available. 1709161 4 62 theme acid 581:584 arg1 sequences 586:594 The complete amino acid sequences 562:594 The complete amino acid sequences derived from the cloned cDNAs 562:624 The complete amino acid sequences derived from the cloned cDNAs represent two new IGFBPs. 1709161 7 63 theme other 923:927 arg1 BPs 941:943 the other three human BPs 919:943 the other three human BPs 919:943 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 1 64 theme molecular 204:212 arg1 nonreduced 247:256 nonreduced 247:256 nonreduced 247:256 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 64 theme molecular 204:212 arg1 masses 214:219 apparent molecular masses 195:219 apparent molecular masses of 24, 28-30, and 30 kDa 195:244 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 9 65 from region 1109:1114 arg1 located 1081:1087 located 1081:1087 located 1081:1087 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 7 66 theme pronounced 896:905 arg1 homology 907:914 pronounced homology 896:914 pronounced homology to the other three human BPs 896:943 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 1 67 with proteins 173:180 arg1 nonreduced 247:256 nonreduced 247:256 nonreduced 247:256 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 67 with proteins 173:180 arg1 masses 214:219 apparent molecular masses 195:219 apparent molecular masses of 24, 28-30, and 30 kDa 195:244 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 7 68 theme -5 885:886 arg1 NH2- 843:846 NH2- 843:846 NH2- 843:846 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 7 68 theme -5 885:886 arg1 thirds 866:871 COOH-terminal thirds 852:871 COOH-terminal thirds 852:871 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 9 69 theme acid 1066:1069 arg1 -5 1148:1149 -5 1148:1149 -5 1148:1149 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 69 theme acid 1066:1069 arg1 -3 1140:1141 -3 1140:1141 -3 1140:1141 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 69 theme acid 1066:1069 arg1 positions 1071:1079 10 amino acid positions 1057:1079 10 amino acid positions located in the NH2-terminal region and shared by BP-1 1057:1133 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 69 theme acid 1066:1069 arg1 -2 1136:1137 -2 1136:1137 -2 1136:1137 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 0 70 theme growth 68:73 arg1 factor 75:80 30-kDa insulin-like growth factor 48:80 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 9 71 theme located 1081:1087 arg1 -5 1148:1149 -5 1148:1149 -5 1148:1149 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 71 theme located 1081:1087 arg1 -3 1140:1141 -3 1140:1141 -3 1140:1141 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 71 theme located 1081:1087 arg1 positions 1071:1079 10 amino acid positions 1057:1079 10 amino acid positions located in the NH2-terminal region and shared by BP-1 1057:1133 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 71 theme located 1081:1087 arg1 -2 1136:1137 -2 1136:1137 -2 1136:1137 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 7 72 theme COOH-terminal 852:864 arg1 thirds 866:871 COOH-terminal thirds 852:871 COOH-terminal thirds 852:871 The NH2- and COOH-terminal thirds of BP-4 and -5 display pronounced homology to the other three human BPs. 1709161 1 73 theme insulin-like 138:149 arg1 IGFBP 183:187 IGFBP 183:187 IGFBP 183:187 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 73 theme insulin-like 138:149 arg1 proteins 173:180 Three insulin-like growth factor binding proteins 132:180 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, 132:257 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 6 74 contain contain 736:742 arg2 proteins 766:773 the mature proteins 755:773 the mature proteins 755:773 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 74 contain contain 736:742 arg2 21 751:752 21 751:752 21 751:752 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 74 contain contain 736:742 arg1 sequences 711:719 The prepeptide sequences 696:719 The prepeptide sequences of BP-4 and -5 696:734 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 6 74 contain contain 736:742 arg2 27 744:745 27 744:745 27 744:745 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 0 75 theme binding 82:88 arg1 proteins 90:97 two new 30-kDa insulin-like growth factor binding proteins 40:97 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 1 76 theme factor 158:163 arg1 IGFBP 183:187 IGFBP 183:187 IGFBP 183:187 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 1 76 theme factor 158:163 arg1 proteins 173:180 Three insulin-like growth factor binding proteins 132:180 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, 132:257 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 10 77 theme preferential 1212:1223 arg1 affinity 1225:1232 the preferential affinity 1208:1232 the preferential affinity of BP-4 for IGF II 1208:1251 These differences may account for the preferential affinity of BP-4 for IGF II. 1709161 11 78 theme intriguing 1261:1270 arg1 homology 1272:1279 A most intriguing homology 1254:1279 A most intriguing homology 1254:1279 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 13 79 from role 1661:1664 arg1 metabolism 1680:1689 metabolism 1680:1689 metabolism 1680:1689 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 13 79 from role 1661:1664 arg1 growth 1669:1674 growth 1669:1674 growth 1669:1674 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 13 80 theme in 1628:1629 arg1 studies 1636:1642 in vivo studies 1628:1642 in vivo studies 1628:1642 They will allow their expression and production in sufficient quantities for in vivo studies to unravel their role in growth and metabolism. 1709161 0 81 theme adult 113:117 arg1 serum 125:129 adult human serum 113:129 adult human serum 113:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 9 82 theme shared 1120:1125 arg1 -5 1148:1149 -5 1148:1149 -5 1148:1149 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 82 theme shared 1120:1125 arg1 -3 1140:1141 -3 1140:1141 -3 1140:1141 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 82 theme shared 1120:1125 arg1 positions 1071:1079 10 amino acid positions 1057:1079 10 amino acid positions located in the NH2-terminal region and shared by BP-1 1057:1133 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 9 82 theme shared 1120:1125 arg1 -2 1136:1137 -2 1136:1137 -2 1136:1137 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 2 83 theme binding 342:348 arg1 protein 350:356 the 24-kDa binding protein 331:356 the 24-kDa binding protein 331:356 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 2 84 theme 30-kDa 390:395 arg1 BP 397:398 the 30-kDa BP 386:398 the 30-kDa BP 386:398 The 15 NH2-terminal amino acids of the 24-kDa binding protein are identical with those of the 30-kDa BP. 1709161 11 85 theme class 1464:1468 arg1 antigen 1492:1498 class II histocompatibility antigen 1464:1498 the class II histocompatibility antigen 1460:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 3 86 theme molecular 414:422 arg1 mass 424:427 The apparent molecular mass 401:427 The apparent molecular mass of the latter 401:441 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 3 87 theme 24-kDa 551:556 arg1 BP 558:559 the isolated 24-kDa BP 538:559 the isolated 24-kDa BP 538:559 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 0 88 theme 30-kDa 48:53 arg1 factor 75:80 30-kDa insulin-like growth factor 48:80 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 9 89 from BP-4 1168:1171 arg1 different 1155:1163 different 1155:1163 different 1155:1163 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 3 90 theme latter 436:441 arg1 mass 424:427 The apparent molecular mass 401:427 The apparent molecular mass of the latter 401:441 The apparent molecular mass of the latter is reduced to 24 kDa by N-glycanase, suggesting that the 30-kDa BP is the glycosylated form of the isolated 24-kDa BP. 1709161 1 91 theme human 283:287 arg1 serum 289:293 human serum 283:293 human serum 283:293 Three insulin-like growth factor binding proteins (IGFBP) with apparent molecular masses of 24, 28-30, and 30 kDa, nonreduced, have been isolated from human serum. 1709161 9 92 from different 1155:1163 arg1 BP-4 1168:1171 BP-4 1168:1171 BP-4 1168:1171 10 amino acid positions located in the NH2-terminal region and shared by BP-1, -2, -3, and -5 are different in BP-4. 1709161 11 93 theme histocompatibility 1473:1490 arg1 antigen 1492:1498 class II histocompatibility antigen 1464:1498 the class II histocompatibility antigen 1460:1498 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 1709161 6 94 theme -5 733:734 arg1 sequences 711:719 The prepeptide sequences 696:719 The prepeptide sequences of BP-4 and -5 696:734 The prepeptide sequences of BP-4 and -5 contain 27 and 21, the mature proteins 213 and 237 amino acids, respectively (Mr = 22,610 and 25,980). 1709161 0 95 attach isolated 99:106 arg2 proteins 90:97 two new 30-kDa insulin-like growth factor binding proteins 40:97 two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum 40:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 0 95 attach isolated 99:106 arg1 serum 125:129 adult human serum 113:129 adult human serum 113:129 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. 1709161 11 96 theme human 1367:1371 arg1 thyroglobulin 1373:1385 human thyroglobulin 1367:1385 human thyroglobulin 1367:1385 A most intriguing homology exists between the COOH-terminal quarter of the five IGFBPs, 10 repetitive domains of human thyroglobulin, a gastrointestinal tumor-associated antigen, and the invariant chain of the class II histocompatibility antigen. 28334865 7 0 theme Notch1 1474:1479 arg1 signaling 1481:1489 Notch1 signaling 1474:1489 Notch1 signaling 1474:1489 The reported Dowling-Degos mutations of POFUT1, except for M262T, fail to rescue Notch1 signaling efficiently in the CRISPR-engineered POFUT1-/- background. 28334865 6 1 theme side 1294:1297 arg1 chains 1299:1304 the side chains 1290:1304 the side chains of R43 and D244 1290:1320 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 7 2 theme POFUT1 1433:1438 arg1 mutations 1420:1428 The reported Dowling-Degos mutations 1393:1428 The reported Dowling-Degos mutations 1393:1428 The reported Dowling-Degos mutations of POFUT1, except for M262T, fail to rescue Notch1 signaling efficiently in the CRISPR-engineered POFUT1-/- background. 28334865 1 3 theme epidermal 270:278 arg1 repeats 299:305 epidermal growth factor-like repeats 270:305 epidermal growth factor-like repeats of recipient proteins 270:327 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 4 4 theme oncogenic 871:879 arg1 signaling 881:889 Normal and oncogenic signaling 860:889 Normal and oncogenic signaling 860:889 Normal and oncogenic signaling are rescued by wild-type POFUT1 but rescue is impaired by an active-site R240A mutation. 28334865 3 5 theme U2OS 721:724 arg1 cells 726:730 U2OS cells 721:730 U2OS cells 721:730 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 7 6 theme reported 1397:1404 arg1 mutations 1420:1428 The reported Dowling-Degos mutations 1393:1428 The reported Dowling-Degos mutations 1393:1428 The reported Dowling-Degos mutations of POFUT1, except for M262T, fail to rescue Notch1 signaling efficiently in the CRISPR-engineered POFUT1-/- background. 28334865 3 7 from cells 726:730 arg1 knockout 699:706 knockout 699:706 knockout 699:706 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 6 8 theme backbone 1249:1256 arg1 movement 1273:1280 limited backbone conformational movement 1241:1280 limited backbone conformational movement 1241:1280 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 2 9 theme POFUT1 563:568 arg1 structures 543:552 the structures 539:552 the structures of human POFUT1 in free and GDP-fucose bound states 539:604 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 3 10 theme normal 748:753 arg1 signaling 762:770 normal Notch1 signaling 748:770 normal Notch1 signaling 748:770 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 5 11 theme overall 1092:1098 arg1 RMSD 1109:1112 an overall backbone RMSD 1089:1112 an overall backbone RMSD of 0.93 Å 1089:1122 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 4 12 theme R240A 964:968 arg1 mutation 970:977 an active-site R240A mutation 949:977 an active-site R240A mutation 949:977 Normal and oncogenic signaling are rescued by wild-type POFUT1 but rescue is impaired by an active-site R240A mutation. 28334865 6 13 theme limited 1241:1247 arg1 movement 1273:1280 limited backbone conformational movement 1241:1280 limited backbone conformational movement 1241:1280 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 3 14 theme ligand-independent 781:798 arg1 signaling 800:808 the ligand-independent signaling 777:808 the ligand-independent signaling associated with leukemogenic mutations of Notch1 777:857 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 0 15 theme mutations 132:140 arg1 requirement 31:41 its requirement 27:41 its requirement in ligand-independent oncogenic Notch signaling 27:89 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 0 15 theme mutations 132:140 arg1 effects 107:113 functional effects 96:113 functional effects of Dowling-Degos mutations 96:140 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 0 16 theme POFUT1 19:24 arg1 Structure 0:8 Structure 0:8 Structure of human POFUT1 0:24 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 1 17 theme acceptor 219:226 arg1 sites 228:232 acceptor sites 219:232 acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins 219:327 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 5 18 theme mature 1178:1183 arg1 protein 1185:1191 the mature protein 1174:1191 the mature protein 1174:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 6 19 theme R43 1309:1311 arg1 chains 1299:1304 the side chains 1290:1304 the side chains of R43 and D244 1290:1320 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 2 20 theme free 573:576 arg1 states 599:604 free and GDP-fucose bound states 573:604 free and GDP-fucose bound states 573:604 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 3 21 theme leukemogenic 826:837 arg1 mutations 839:847 leukemogenic mutations 826:847 leukemogenic mutations of Notch1 826:857 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 2 22 theme human 518:522 arg1 Notch1 524:529 human Notch1 518:529 human Notch1 518:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 0 23 theme ligand-independent 46:63 arg1 signaling 81:89 ligand-independent oncogenic Notch signaling 46:89 ligand-independent oncogenic Notch signaling 46:89 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 5 24 theme primary 1133:1139 arg1 identity 1150:1157 primary sequence identity 1133:1157 primary sequence identity of only 39% in the mature protein 1133:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 0 25 theme Notch 75:79 arg1 signaling 81:89 ligand-independent oncogenic Notch signaling 46:89 ligand-independent oncogenic Notch signaling 46:89 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 1 26 theme Notch 347:351 arg1 transduction 360:371 Notch signal transduction 347:371 Notch signal transduction in mammals 347:382 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 2 27 theme bound 593:597 arg1 states 599:604 free and GDP-fucose bound states 573:604 free and GDP-fucose bound states 573:604 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 5 28 theme 0.93 1117:1120 arg1 RMSD 1109:1112 an overall backbone RMSD 1089:1112 an overall backbone RMSD of 0.93 Å 1089:1122 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 0 29 from requirement 31:41 arg1 signaling 81:89 ligand-independent oncogenic Notch signaling 46:89 ligand-independent oncogenic Notch signaling 46:89 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 2 30 theme Dowling-Degos 633:645 arg1 mutations 647:655 Dowling-Degos mutations 633:655 Dowling-Degos mutations 633:655 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 0 31 theme functional 96:105 arg1 effects 107:113 functional effects 96:113 functional effects of Dowling-Degos mutations 96:140 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 5 32 theme human 1009:1013 arg1 enzyme 1015:1020 the human enzyme 1005:1020 the human enzyme 1005:1020 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 2 33 theme oncogenic 441:449 arg1 signaling 451:459 the oncogenic signaling 437:459 the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1 437:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 1 34 theme growth 280:285 arg1 repeats 299:305 epidermal growth factor-like repeats 270:305 epidermal growth factor-like repeats of recipient proteins 270:327 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 2 35 theme POFUT1 666:671 arg1 function 673:680 human POFUT1 function 660:680 human POFUT1 function 660:680 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 2 36 theme POFUT1 422:427 arg1 loss 429:432 POFUT1 loss 422:432 POFUT1 loss 422:432 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 0 37 theme human 13:17 arg1 POFUT1 19:24 human POFUT1 13:24 human POFUT1 13:24 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 1 38 theme repeats 299:305 arg1 serine 237:242 serine 237:242 serine 237:242 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 1 38 theme repeats 299:305 arg1 threonine 248:256 threonine 248:256 threonine 248:256 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 5 39 theme overall 984:990 arg1 structure 992:1000 The overall structure 980:1000 The overall structure of the human enzyme 980:1020 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 2 40 theme certain 477:483 arg1 mutations 505:513 certain leukemia-associated mutations 477:513 certain leukemia-associated mutations of human Notch1 477:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 1 41 theme recipient 310:318 arg1 proteins 320:327 recipient proteins 310:327 recipient proteins 310:327 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 7 42 theme CRISPR-engineered 1510:1526 arg1 background 1538:1547 the CRISPR-engineered POFUT1-/- background 1506:1547 the CRISPR-engineered POFUT1-/- background 1506:1547 The reported Dowling-Degos mutations of POFUT1, except for M262T, fail to rescue Notch1 signaling efficiently in the CRISPR-engineered POFUT1-/- background. 28334865 0 43 dep requirement 31:41 arg1 Structure 0:8 Structure 0:8 Structure of human POFUT1 0:24 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 4 44 theme Normal 860:865 arg1 signaling 881:889 Normal and oncogenic signaling 860:889 Normal and oncogenic signaling 860:889 Normal and oncogenic signaling are rescued by wild-type POFUT1 but rescue is impaired by an active-site R240A mutation. 28334865 5 45 theme % 1169:1169 arg1 identity 1150:1157 primary sequence identity 1133:1157 primary sequence identity of only 39% in the mature protein 1133:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 5 46 theme elegans 1067:1073 arg1 protein 1075:1081 the Caenorhabditis elegans protein 1048:1081 the Caenorhabditis elegans protein 1048:1081 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 2 47 from effects 622:628 arg1 function 673:680 human POFUT1 function 660:680 human POFUT1 function 660:680 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 2 48 from consequences 406:417 arg1 signaling 451:459 the oncogenic signaling 437:459 the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1 437:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 2 49 from structures 543:552 arg1 states 599:604 free and GDP-fucose bound states 573:604 free and GDP-fucose bound states 573:604 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 5 50 from % 1169:1169 arg1 protein 1185:1191 the mature protein 1174:1191 the mature protein 1174:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 5 51 theme sequence 1141:1148 arg1 identity 1150:1157 primary sequence identity 1133:1157 primary sequence identity of only 39% in the mature protein 1133:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 4 52 theme wild-type 906:914 arg1 POFUT1 916:921 wild-type POFUT1 906:921 wild-type POFUT1 906:921 Normal and oncogenic signaling are rescued by wild-type POFUT1 but rescue is impaired by an active-site R240A mutation. 28334865 8 53 theme Notch1 1634:1639 arg1 mutations 1641:1649 Notch1 mutations 1634:1649 Notch1 mutations 1634:1649 Together, these studies identify POFUT1 as a potential target for cancers driven by Notch1 mutations and provide a structural roadmap for its inhibition. 28334865 5 54 from protein 1185:1191 arg1 identity 1150:1157 primary sequence identity 1133:1157 primary sequence identity of only 39% in the mature protein 1133:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 6 55 theme conformational 1258:1271 arg1 movement 1273:1280 limited backbone conformational movement 1241:1280 limited backbone conformational movement 1241:1280 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 7 56 theme Dowling-Degos 1406:1418 arg1 mutations 1420:1428 The reported Dowling-Degos mutations 1393:1428 The reported Dowling-Degos mutations 1393:1428 The reported Dowling-Degos mutations of POFUT1, except for M262T, fail to rescue Notch1 signaling efficiently in the CRISPR-engineered POFUT1-/- background. 28334865 0 57 theme Dowling-Degos 118:130 arg1 mutations 132:140 Dowling-Degos mutations 118:140 Dowling-Degos mutations 118:140 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 1 58 theme fucose 200:205 arg1 residues 207:214 fucose residues 200:214 fucose residues 200:214 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 2 59 theme human 557:561 arg1 POFUT1 563:568 human POFUT1 557:568 human POFUT1 557:568 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 1 60 dep serine 237:242 arg1 residues 258:265 residues 258:265 residues 258:265 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 3 61 from knockout 699:706 arg1 cells 726:730 U2OS cells 721:730 U2OS cells 721:730 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 3 62 theme Notch1 755:760 arg1 signaling 762:770 normal Notch1 signaling 748:770 normal Notch1 signaling 748:770 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 5 63 theme backbone 1100:1107 arg1 RMSD 1109:1112 an overall backbone RMSD 1089:1112 an overall backbone RMSD of 0.93 Å 1089:1122 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 2 64 dep bound 593:597 arg1 GDP-fucose 582:591 GDP-fucose 582:591 GDP-fucose 582:591 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 4 65 theme active-site 952:962 arg1 mutation 970:977 an active-site R240A mutation 949:977 an active-site R240A mutation 949:977 Normal and oncogenic signaling are rescued by wild-type POFUT1 but rescue is impaired by an active-site R240A mutation. 28334865 6 66 theme D244 1317:1320 arg1 chains 1299:1304 the side chains 1290:1304 the side chains of R43 and D244 1290:1320 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 6 67 theme direct 1339:1344 arg1 contact 1346:1352 direct contact 1339:1352 direct contact with the fucose moiety in the complex 1339:1390 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 0 68 from effects 107:113 arg1 signaling 81:89 ligand-independent oncogenic Notch signaling 46:89 ligand-independent oncogenic Notch signaling 46:89 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 6 69 theme human 1220:1224 arg1 enzyme 1226:1231 the human enzyme 1216:1231 the human enzyme 1216:1231 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 1 70 from sites 228:232 arg1 serine 237:242 serine 237:242 serine 237:242 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 1 70 from sites 228:232 arg1 threonine 248:256 threonine 248:256 threonine 248:256 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 0 71 theme oncogenic 65:73 arg1 signaling 81:89 ligand-independent oncogenic Notch signaling 46:89 ligand-independent oncogenic Notch signaling 46:89 Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. 28334865 6 72 with contact 1346:1352 arg1 moiety 1370:1375 the fucose moiety 1359:1375 the fucose moiety in the complex 1359:1390 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 1 73 theme Protein 143:149 arg1 O-fucosyltransferase-1  151:173 Protein O-fucosyltransferase-1  143:173 Protein O-fucosyltransferase-1 (POFUT1) 143:181 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 1 73 theme Protein 143:149 arg1 POFUT1 175:180 POFUT1 175:180 POFUT1 175:180 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 6 74 from moiety 1370:1375 arg1 complex 1384:1390 the complex 1380:1390 the complex 1380:1390 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 2 75 theme leukemia-associated 485:503 arg1 mutations 505:513 certain leukemia-associated mutations 477:513 certain leukemia-associated mutations of human Notch1 477:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 2 76 theme human 660:664 arg1 function 673:680 human POFUT1 function 660:680 human POFUT1 function 660:680 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 6 77 theme fucose 1363:1368 arg1 moiety 1370:1375 the fucose moiety 1359:1375 the fucose moiety in the complex 1359:1390 GDP-fucose binding to the human enzyme induces limited backbone conformational movement, though the side chains of R43 and D244 reorient to make direct contact with the fucose moiety in the complex. 28334865 1 78 theme signal 353:358 arg1 transduction 360:371 Notch signal transduction 347:371 Notch signal transduction in mammals 347:382 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 2 79 theme mutations 647:655 arg1 effects 622:628 the effects 618:628 the effects of Dowling-Degos mutations on human POFUT1 function 618:680 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 3 80 theme Notch1 852:857 arg1 mutations 839:847 leukemogenic mutations 826:847 leukemogenic mutations of Notch1 826:857 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 28334865 2 81 theme Notch1 524:529 arg1 mutations 505:513 certain leukemia-associated mutations 477:513 certain leukemia-associated mutations of human Notch1 477:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 8 82 theme potential 1595:1603 arg1 POFUT1 1583:1588 POFUT1 1583:1588 POFUT1 1583:1588 Together, these studies identify POFUT1 as a potential target for cancers driven by Notch1 mutations and provide a structural roadmap for its inhibition. 28334865 8 82 theme potential 1595:1603 arg1 target 1605:1610 a potential target 1593:1610 a potential target for cancers driven by Notch1 mutations 1593:1649 Together, these studies identify POFUT1 as a potential target for cancers driven by Notch1 mutations and provide a structural roadmap for its inhibition. 28334865 1 83 from transduction 360:371 arg1 mammals 376:382 mammals 376:382 mammals 376:382 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 1 84 theme factor-like 287:297 arg1 repeats 299:305 epidermal growth factor-like repeats 270:305 epidermal growth factor-like repeats of recipient proteins 270:327 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 2 85 theme loss 429:432 arg1 consequences 406:417 the consequences 402:417 the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1 402:529 Here, we examine the consequences of POFUT1 loss on the oncogenic signaling associated with certain leukemia-associated mutations of human Notch1, report the structures of human POFUT1 in free and GDP-fucose bound states, and assess the effects of Dowling-Degos mutations on human POFUT1 function. 28334865 5 86 from identity 1150:1157 arg1 protein 1185:1191 the mature protein 1174:1191 the mature protein 1174:1191 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 1 87 theme proteins 320:327 arg1 repeats 299:305 epidermal growth factor-like repeats 270:305 epidermal growth factor-like repeats of recipient proteins 270:327 Protein O-fucosyltransferase-1 (POFUT1), which transfers fucose residues to acceptor sites on serine and threonine residues of epidermal growth factor-like repeats of recipient proteins, is essential for Notch signal transduction in mammals. 28334865 7 88 theme POFUT1-/- 1528:1536 arg1 background 1538:1547 the CRISPR-engineered POFUT1-/- background 1506:1547 the CRISPR-engineered POFUT1-/- background 1506:1547 The reported Dowling-Degos mutations of POFUT1, except for M262T, fail to rescue Notch1 signaling efficiently in the CRISPR-engineered POFUT1-/- background. 28334865 5 89 theme Caenorhabditis 1052:1065 arg1 protein 1075:1081 the Caenorhabditis elegans protein 1048:1081 the Caenorhabditis elegans protein 1048:1081 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 5 90 theme enzyme 1015:1020 arg1 structure 992:1000 The overall structure 980:1000 The overall structure of the human enzyme 980:1020 The overall structure of the human enzyme closely resembles that of the Caenorhabditis elegans protein, with an overall backbone RMSD of 0.93 Å, despite primary sequence identity of only 39% in the mature protein. 28334865 8 91 theme structural 1665:1674 arg1 roadmap 1676:1682 a structural roadmap 1663:1682 a structural roadmap for its inhibition 1663:1701 Together, these studies identify POFUT1 as a potential target for cancers driven by Notch1 mutations and provide a structural roadmap for its inhibition. 28334865 3 92 theme POFUT1 711:716 arg1 knockout 699:706 knockout 699:706 knockout 699:706 CRISPR-mediated knockout of POFUT1 in U2OS cells suppresses both normal Notch1 signaling, and the ligand-independent signaling associated with leukemogenic mutations of Notch1. 12391027 0 0 theme layered 55:61 arg1 fold 63:66 a novel layered fold 47:66 a novel layered fold 47:66 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 4 1 theme superfamily 922:932 arg1 members 903:909 the diverse members 891:909 the diverse members of the TSR superfamily 891:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 2 theme domain 799:804 arg1 This 738:741 This 738:741 This 738:741 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 2 theme domain 799:804 arg1 structure 780:788 the first high-resolution crystal structure 746:788 the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily 746:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 1 3 theme glycosaminoglycan 194:210 arg1 binding 212:218 glycosaminoglycan binding 194:218 glycosaminoglycan binding 194:218 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 0 4 theme novel 49:53 arg1 fold 63:66 a novel layered fold 47:66 a novel layered fold 47:66 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 1 5 theme matrix 290:295 arg1 metalloproteinases 297:314 matrix metalloproteinases 290:314 matrix metalloproteinases 290:314 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 4 6 theme TSR 795:797 arg1 domain 799:804 a TSR domain 793:804 a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily 793:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 2 7 from strands 523:529 arg1 residues 498:505 tryptophan and arginine residues 474:505 tryptophan and arginine residues from respective strands 474:529 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 2 7 from strands 523:529 arg1 layers 464:469 stacked layers 456:469 stacked layers 456:469 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 1 8 theme metalloproteinases 297:314 arg1 attachment 182:191 cell attachment 177:191 cell attachment 177:191 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 8 theme metalloproteinases 297:314 arg1 binding 212:218 glycosaminoglycan binding 194:218 glycosaminoglycan binding 194:218 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 8 theme metalloproteinases 297:314 arg1 inhibition 276:285 inhibition 276:285 inhibition of matrix metalloproteinases 276:314 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 8 theme metalloproteinases 297:314 arg1 activation 249:258 activation 249:258 activation of TGFbeta 249:269 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 8 theme metalloproteinases 297:314 arg1 inhibition 221:230 inhibition 221:230 inhibition of angiogenesis 221:246 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 2 9 theme tryptophan 474:483 arg1 residues 498:505 tryptophan and arginine residues 474:505 tryptophan and arginine residues from respective strands 474:529 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 2 10 theme disulfide 542:550 arg1 bonds 552:556 disulfide bonds 542:556 disulfide bonds 542:556 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 3 11 with interactions 703:714 arg1 ligands 729:735 various ligands 721:735 various ligands 721:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 12 dep spiral 621:626 arg1 charged 640:646 charged 640:646 charged 640:646 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 0 13 theme biological 76:85 arg1 implication 87:97 its biological implication 72:97 its biological implication 72:97 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 1 14 contain contains 125:132 arg1 Thrombospondin-1 100:115 Thrombospondin-1 100:115 Thrombospondin-1 (TSP-1) 100:123 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 14 contain contains 125:132 arg2 TSRs 156:159 TSRs 156:159 TSRs 156:159 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 14 contain contains 125:132 arg1 TSP-1 118:122 TSP-1 118:122 TSP-1 118:122 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 14 contain contains 125:132 arg2 repeats 147:153 three type 1 repeats 134:153 three type 1 repeats (TSRs) 134:160 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 2 15 dep novel 386:390 arg1 three-stranded 407:420 three-stranded 407:420 three-stranded 407:420 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 2 15 dep novel 386:390 arg1 antiparallel 393:404 antiparallel 393:404 antiparallel 393:404 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 1 16 theme type 140:143 arg1 repeats 147:153 three type 1 repeats 134:153 three type 1 repeats (TSRs) 134:160 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 16 theme type 140:143 arg1 TSRs 156:159 TSRs 156:159 TSRs 156:159 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 4 17 theme diverse 895:901 arg1 members 903:909 the diverse members 891:909 the diverse members of the TSR superfamily 891:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 18 theme crystal 772:778 arg1 This 738:741 This 738:741 This 738:741 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 18 theme crystal 772:778 arg1 structure 780:788 the first high-resolution crystal structure 746:788 the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily 746:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 2 19 theme TSRs 346:349 arg1 structure 329:337 The crystal structure 317:337 The crystal structure of the TSRs reported in this article 317:374 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 4 20 theme TSR 918:920 arg1 superfamily 922:932 the TSR superfamily 914:932 the TSR superfamily 914:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 0 21 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 0:98 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 3 22 theme " 685:685 arg1 face 687:690 the "recognition" face 669:690 the "recognition" face 669:690 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 22 theme " 685:685 arg1 groove 648:653 a right-handed spiral, positively charged groove 606:653 a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands 606:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 2 23 theme novel 386:390 arg1 fold 422:425 a novel, antiparallel, three-stranded fold 384:425 a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end 384:568 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 1 24 theme angiogenesis 235:246 arg1 attachment 182:191 cell attachment 177:191 cell attachment 177:191 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 24 theme angiogenesis 235:246 arg1 binding 212:218 glycosaminoglycan binding 194:218 glycosaminoglycan binding 194:218 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 24 theme angiogenesis 235:246 arg1 inhibition 276:285 inhibition 276:285 inhibition of matrix metalloproteinases 276:314 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 24 theme angiogenesis 235:246 arg1 activation 249:258 activation 249:258 activation of TGFbeta 249:269 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 24 theme angiogenesis 235:246 arg1 inhibition 221:230 inhibition 221:230 inhibition of angiogenesis 221:246 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 2 25 theme respective 512:521 arg1 strands 523:529 respective strands 512:529 respective strands 512:529 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 4 26 theme first 750:754 arg1 This 738:741 This 738:741 This 738:741 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 26 theme first 750:754 arg1 structure 780:788 the first high-resolution crystal structure 746:788 the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily 746:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 27 theme structural 850:859 arg1 exploration 876:886 structural and functional exploration 850:886 structural and functional exploration of the diverse members of the TSR superfamily 850:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 0 28 theme type 31:34 arg1 repeats 38:44 the TSP-1 type 1 repeats 21:44 the TSP-1 type 1 repeats 21:44 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 3 29 theme recognition 674:684 arg1 face 687:690 the "recognition" face 669:690 the "recognition" face 669:690 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 29 theme recognition 674:684 arg1 groove 648:653 a right-handed spiral, positively charged groove 606:653 a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands 606:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 2 30 theme residues 498:505 arg1 layers 464:469 stacked layers 456:469 stacked layers 456:469 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 3 31 theme TSR 593:595 arg1 face 581:584 The front face 571:584 The front face of the TSR 571:595 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 0 32 theme TSP-1 25:29 arg1 repeats 38:44 the TSP-1 type 1 repeats 21:44 the TSP-1 type 1 repeats 21:44 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 2 33 theme arginine 489:496 arg1 residues 498:505 tryptophan and arginine residues 474:505 tryptophan and arginine residues from respective strands 474:529 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 0 34 theme repeats 38:44 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 0:98 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 1 35 theme TGFbeta 263:269 arg1 attachment 182:191 cell attachment 177:191 cell attachment 177:191 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 35 theme TGFbeta 263:269 arg1 binding 212:218 glycosaminoglycan binding 194:218 glycosaminoglycan binding 194:218 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 35 theme TGFbeta 263:269 arg1 inhibition 276:285 inhibition 276:285 inhibition of matrix metalloproteinases 276:314 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 35 theme TGFbeta 263:269 arg1 activation 249:258 activation 249:258 activation of TGFbeta 249:269 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 1 35 theme TGFbeta 263:269 arg1 inhibition 221:230 inhibition 221:230 inhibition of angiogenesis 221:246 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 4 36 theme prototypic 822:831 arg1 architecture 833:844 a prototypic architecture 820:844 a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily 820:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 37 theme high-resolution 756:770 arg1 This 738:741 This 738:741 This 738:741 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 37 theme high-resolution 756:770 arg1 structure 780:788 the first high-resolution crystal structure 746:788 the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily 746:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 4 38 theme functional 865:874 arg1 exploration 876:886 structural and functional exploration 850:886 structural and functional exploration of the diverse members of the TSR superfamily 850:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 3 39 theme right-handed 608:619 arg1 groove 648:653 a right-handed spiral, positively charged groove 606:653 a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands 606:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 39 theme right-handed 608:619 arg1 face 687:690 the "recognition" face 669:690 the "recognition" face 669:690 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 40 theme spiral 621:626 arg1 groove 648:653 a right-handed spiral, positively charged groove 606:653 a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands 606:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 40 theme spiral 621:626 arg1 face 687:690 the "recognition" face 669:690 the "recognition" face 669:690 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 4 41 theme members 903:909 arg1 exploration 876:886 structural and functional exploration 850:886 structural and functional exploration of the diverse members of the TSR superfamily 850:932 This is the first high-resolution crystal structure of a TSR domain that provides a prototypic architecture for structural and functional exploration of the diverse members of the TSR superfamily. 12391027 2 42 from layers 464:469 arg1 strands 523:529 respective strands 512:529 respective strands 512:529 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 3 43 theme front 575:579 arg1 face 581:584 The front face 571:584 The front face of the TSR 571:595 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 1 44 theme cell 177:180 arg1 attachment 182:191 cell attachment 177:191 cell attachment 177:191 Thrombospondin-1 (TSP-1) contains three type 1 repeats (TSRs), which mediate cell attachment, glycosaminoglycan binding, inhibition of angiogenesis, activation of TGFbeta, and inhibition of matrix metalloproteinases. 12391027 2 45 theme stacked 456:462 arg1 layers 464:469 stacked layers 456:469 stacked layers 456:469 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 12391027 0 46 dep structure 8:16 arg1 implication 87:97 its biological implication 72:97 its biological implication 72:97 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 0 46 dep structure 8:16 arg1 fold 63:66 a novel layered fold 47:66 a novel layered fold 47:66 Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. 12391027 3 47 contain contains 597:604 arg2 face 687:690 the "recognition" face 669:690 the "recognition" face 669:690 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 47 contain contains 597:604 arg1 face 581:584 The front face 571:584 The front face of the TSR 571:595 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 47 contain contains 597:604 arg2 groove 648:653 a right-handed spiral, positively charged groove 606:653 a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands 606:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 3 48 theme various 721:727 arg1 ligands 729:735 various ligands 721:735 various ligands 721:735 The front face of the TSR contains a right-handed spiral, positively charged groove that might be the "recognition" face, mediating interactions with various ligands. 12391027 2 49 theme crystal 321:327 arg1 structure 329:337 The crystal structure 317:337 The crystal structure of the TSRs reported in this article 317:374 The crystal structure of the TSRs reported in this article reveals a novel, antiparallel, three-stranded fold that consists of alternating stacked layers of tryptophan and arginine residues from respective strands, capped by disulfide bonds on each end. 16763549 7 0 theme physiological 1107:1119 arg1 angiogenesis 1138:1149 physiological and pathological angiogenesis 1107:1149 physiological and pathological angiogenesis 1107:1149 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 2 1 theme muscle 365:370 arg1 SMCs 379:382 SMCs 379:382 SMCs 379:382 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 1 theme muscle 365:370 arg1 cells 372:376 smooth muscle cells 358:376 smooth muscle cells (SMCs) 358:383 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 2 theme smooth 358:363 arg1 SMCs 379:382 SMCs 379:382 SMCs 379:382 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 2 theme smooth 358:363 arg1 cells 372:376 smooth muscle cells 358:376 smooth muscle cells (SMCs) 358:383 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 5 3 theme VEGF 722:725 arg1 binding 727:733 VEGF binding 722:733 VEGF binding 722:733 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 6 4 theme NRP1 867:870 arg1 expression 872:881 NRP1 expression 867:881 NRP1 expression 867:881 Finally, NRP1 expression and its GAG modification post-transcriptionally regulate VEGFR2 protein expression. 16763549 7 5 theme pathological 1125:1136 arg1 angiogenesis 1138:1149 physiological and pathological angiogenesis 1107:1149 physiological and pathological angiogenesis 1107:1149 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 7 6 theme new 1089:1091 arg1 insights 1093:1100 new insights 1089:1100 new insights into physiological and pathological angiogenesis 1089:1149 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 4 7 theme glycosaminoglycan 637:653 arg1 chains 661:666 the NRP1 glycosaminoglycan (GAG) chains 628:666 the NRP1 glycosaminoglycan (GAG) chains 628:666 The composition of the NRP1 glycosaminoglycan (GAG) chains differs between ECs and SMCs. 16763549 6 8 theme protein 947:953 arg1 expression 955:964 VEGFR2 protein expression 940:964 VEGFR2 protein expression 940:964 Finally, NRP1 expression and its GAG modification post-transcriptionally regulate VEGFR2 protein expression. 16763549 7 9 mod modification 1000:1011 arg1 NRP1 1016:1019 NRP1 1016:1019 NRP1 1016:1019 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 7 9 mod modification 1000:1011 arg3 GAG 996:998 GAG modification 996:1011 GAG modification of NRP1 996:1019 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 1 10 theme angiogenic 176:185 arg1 signals 187:193 the angiogenic signals 172:193 the angiogenic signals 172:193 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 0 11 theme Glycosaminoglycan 0:16 arg1 modification 18:29 Glycosaminoglycan modification 0:29 Glycosaminoglycan modification of neuropilin-1 0:45 Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. 16763549 2 12 theme endothelial 330:340 arg1 ECs 349:351 ECs 349:351 ECs 349:351 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 12 theme endothelial 330:340 arg1 cells 342:346 vascular endothelial cells 321:346 vascular endothelial cells (ECs) 321:352 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 0 13 theme neuropilin-1 34:45 arg1 modification 18:29 Glycosaminoglycan modification 0:29 Glycosaminoglycan modification of neuropilin-1 0:45 Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. 16763549 2 14 theme physiological 254:266 arg1 angiogenesis 285:296 physiological and pathological angiogenesis 254:296 physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs) 254:383 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 3 15 theme heparan 543:549 arg1 sulfate 551:557 heparan sulfate 543:557 heparan sulfate 543:557 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 5 16 theme opposite 809:816 arg1 responsiveness 818:831 opposite responsiveness 809:831 opposite responsiveness to VEGF in ECs and SMCs 809:855 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 3 17 theme chondroitin 562:572 arg1 sulfate 574:580 chondroitin sulfate 562:580 chondroitin sulfate 562:580 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 0 18 theme VEGFR2 57:62 arg1 signaling 64:72 VEGFR2 signaling 57:72 VEGFR2 signaling 57:72 Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. 16763549 3 19 theme substantial 477:487 arg1 proteoglycan 509:520 proteoglycan 509:520 proteoglycan modified with either heparan sulfate or chondroitin sulfate 509:580 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 3 19 theme substantial 477:487 arg1 fraction 489:496 a substantial fraction 475:496 a substantial fraction of NRP1 475:504 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 4 20 theme NRP1 632:635 arg1 GAG 656:658 GAG 656:658 GAG 656:658 The composition of the NRP1 glycosaminoglycan (GAG) chains differs between ECs and SMCs. 16763549 4 20 theme NRP1 632:635 arg1 glycosaminoglycan 637:653 NRP1 glycosaminoglycan 632:653 the NRP1 glycosaminoglycan (GAG) chains 628:666 The composition of the NRP1 glycosaminoglycan (GAG) chains differs between ECs and SMCs. 16763549 6 21 theme VEGFR2 940:945 arg1 expression 955:964 VEGFR2 protein expression 940:964 VEGFR2 protein expression 940:964 Finally, NRP1 expression and its GAG modification post-transcriptionally regulate VEGFR2 protein expression. 16763549 7 22 theme VEGF 1057:1060 arg1 signaling 1062:1070 VEGF signaling 1057:1070 VEGF signaling 1057:1070 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 3 23 theme single 587:592 arg1 Ser 604:606 a single conserved Ser 585:606 a single conserved Ser 585:606 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 7 24 theme NRP1 1016:1019 arg1 modification 1000:1011 GAG modification 996:1011 GAG modification of NRP1 996:1019 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 5 25 theme NRP1 795:798 arg1 composition 780:790 the differential GAG composition 759:790 the differential GAG composition of NRP1 759:798 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 5 26 theme differential 763:774 arg1 composition 780:790 the differential GAG composition 759:790 the differential GAG composition of NRP1 759:798 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 1 27 theme vascular 116:123 arg1 VEGF 152:155 VEGF 152:155 VEGF 152:155 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 1 27 theme vascular 116:123 arg1 factor 144:149 vascular endothelial growth factor 116:149 vascular endothelial growth factor (VEGF) 116:156 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 5 28 theme GAG 776:778 arg1 composition 780:790 the differential GAG composition 759:790 the differential GAG composition of NRP1 759:798 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 2 29 theme VEGF 222:225 arg1 signaling 227:235 VEGF signaling 222:235 VEGF signaling 222:235 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 1 30 theme endothelial 125:135 arg1 VEGF 152:155 VEGF 152:155 VEGF 152:155 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 1 30 theme endothelial 125:135 arg1 factor 144:149 vascular endothelial growth factor 116:149 vascular endothelial growth factor (VEGF) 116:156 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 0 31 mod modification 18:29 arg3 Glycosaminoglycan 0:16 Glycosaminoglycan modification 0:29 Glycosaminoglycan modification of neuropilin-1 0:45 Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. 16763549 0 31 mod modification 18:29 arg1 neuropilin-1 34:45 neuropilin-1 34:45 neuropilin-1 34:45 Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. 16763549 2 32 theme pathological 272:283 arg1 angiogenesis 285:296 physiological and pathological angiogenesis 254:296 physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs) 254:383 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 5 33 from responsiveness 818:831 arg1 ECs 844:846 ECs 844:846 ECs 844:846 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 5 33 from responsiveness 818:831 arg1 SMCs 852:855 SMCs 852:855 SMCs 852:855 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 1 34 theme growth 137:142 arg1 VEGF 152:155 VEGF 152:155 VEGF 152:155 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 1 34 theme growth 137:142 arg1 factor 144:149 vascular endothelial growth factor 116:149 vascular endothelial growth factor (VEGF) 116:156 Neuropilin-1 (NRP1) is a co-receptor for vascular endothelial growth factor (VEGF) that enhances the angiogenic signals cooperatively with VEGFR2. 16763549 2 35 theme vascular 321:328 arg1 ECs 349:351 ECs 349:351 ECs 349:351 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 35 theme vascular 321:328 arg1 cells 342:346 vascular endothelial cells 321:346 vascular endothelial cells (ECs) 321:352 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 7 36 theme critical 1029:1036 arg1 role 1038:1041 a critical role 1027:1041 a critical role 1027:1041 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 2 37 from effects 310:316 arg1 SMCs 379:382 SMCs 379:382 SMCs 379:382 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 37 from effects 310:316 arg1 cells 372:376 smooth muscle cells 358:376 smooth muscle cells (SMCs) 358:383 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 37 from effects 310:316 arg1 ECs 349:351 ECs 349:351 ECs 349:351 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 2 37 from effects 310:316 arg1 cells 342:346 vascular endothelial cells 321:346 vascular endothelial cells (ECs) 321:352 VEGF signaling is essential for physiological and pathological angiogenesis through its effects on vascular endothelial cells (ECs) and smooth muscle cells (SMCs), but the mechanisms coordinating this response are not well understood. 16763549 3 38 theme conserved 594:602 arg1 Ser 604:606 a single conserved Ser 585:606 a single conserved Ser 585:606 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 7 39 theme GAG 996:998 arg1 modification 1000:1011 GAG modification 996:1011 GAG modification of NRP1 996:1019 These findings indicate that GAG modification of NRP1 plays a critical role in modulating VEGF signaling, and may provide new insights into physiological and pathological angiogenesis. 16763549 3 40 theme NRP1 501:504 arg1 proteoglycan 509:520 proteoglycan 509:520 proteoglycan modified with either heparan sulfate or chondroitin sulfate 509:580 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 3 40 theme NRP1 501:504 arg1 fraction 489:496 a substantial fraction 475:496 a substantial fraction of NRP1 475:504 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 16763549 5 41 theme cell 743:746 arg1 types 748:752 both cell types 738:752 both cell types 738:752 Glycosylation increased VEGF binding in both cell types, but the differential GAG composition of NRP1 mediates opposite responsiveness to VEGF in ECs and SMCs. 16763549 6 42 theme GAG 891:893 arg1 modification 895:906 its GAG modification 887:906 its GAG modification 887:906 Finally, NRP1 expression and its GAG modification post-transcriptionally regulate VEGFR2 protein expression. 16763549 4 43 theme chains 661:666 arg1 composition 613:623 The composition 609:623 The composition of the NRP1 glycosaminoglycan (GAG) chains 609:666 The composition of the NRP1 glycosaminoglycan (GAG) chains differs between ECs and SMCs. 16763549 3 44 from proteoglycan 509:520 arg1 Ser 604:606 a single conserved Ser 585:606 a single conserved Ser 585:606 Here we show that a substantial fraction of NRP1 is proteoglycan modified with either heparan sulfate or chondroitin sulfate on a single conserved Ser. 17051150 6 0 theme pathway 1170:1176 arg1 convertases 1178:1188 the alternative pathway convertases 1154:1188 the alternative pathway convertases 1154:1188 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 0 1 with complex 20:26 arg1 CRIg 33:36 CRIg 33:36 CRIg 33:36 Structure of C3b in complex with CRIg gives insights into regulation of complement activation. 17051150 4 2 dep present 687:693 arg1 provide 770:776 provide 770:776 provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement 770:857 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 4 2 dep present 687:693 arg1 using 750:754 using 750:754 using CRIg mutants 750:767 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 2 3 theme convertases 407:417 arg1 formation 370:378 formation 370:378 formation of the alternative pathway convertases 370:417 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 6 4 theme alternative 1158:1168 arg1 convertases 1178:1188 the alternative pathway convertases 1154:1188 the alternative pathway convertases 1154:1188 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 3 5 theme complement 428:437 arg1 CRIg 420:423 CRIg 420:423 CRIg 420:423 CRIg, a complement receptor expressed on macrophages, binds to C3b and iC3b mediating phagocytosis of the particles, but it is unknown how CRIg selectively recognizes proteolytic C3-fragments and whether binding of CRIg to C3b inhibits convertase activation. 17051150 3 5 theme complement 428:437 arg1 receptor 439:446 a complement receptor 426:446 a complement receptor expressed on macrophages 426:471 CRIg, a complement receptor expressed on macrophages, binds to C3b and iC3b mediating phagocytosis of the particles, but it is unknown how CRIg selectively recognizes proteolytic C3-fragments and whether binding of CRIg to C3b inhibits convertase activation. 17051150 0 6 theme activation 83:92 arg1 regulation 58:67 regulation 58:67 regulation of complement activation 58:92 Structure of C3b in complex with CRIg gives insights into regulation of complement activation. 17051150 0 7 from Structure 0:8 arg1 complex 20:26 complex 20:26 complex with CRIg 20:36 Structure of C3b in complex with CRIg gives insights into regulation of complement activation. 17051150 4 8 from structure 707:715 arg1 complex 727:733 complex 727:733 complex with CRIg 727:743 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 0 9 theme complement 72:81 arg1 activation 83:92 complement activation 72:92 complement activation 72:92 Structure of C3b in complex with CRIg gives insights into regulation of complement activation. 17051150 4 10 theme crystal 699:705 arg1 structure 707:715 the crystal structure 695:715 the crystal structure of C3b in complex with CRIg 695:743 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 7 11 theme complex 1232:1238 arg1 rearrangements 1266:1279 the complex macromolecular structural rearrangements 1228:1279 the complex macromolecular structural rearrangements 1228:1279 The structure provides insights into the complex macromolecular structural rearrangements that occur during complement activation and inhibition. 17051150 1 12 theme innate 138:143 arg1 system 152:157 the innate immune system 134:157 the innate immune system 134:157 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 4 13 theme complement 848:857 arg1 pathway 837:843 the alternative pathway 821:843 the alternative pathway of complement 821:857 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 3 14 theme CRIg 635:638 arg1 binding 624:630 binding 624:630 binding of CRIg to C3b 624:645 CRIg, a complement receptor expressed on macrophages, binds to C3b and iC3b mediating phagocytosis of the particles, but it is unknown how CRIg selectively recognizes proteolytic C3-fragments and whether binding of CRIg to C3b inhibits convertase activation. 17051150 6 15 theme potent 1134:1139 arg1 inhibitor 1141:1149 a potent inhibitor 1132:1149 a potent inhibitor of the alternative pathway convertases 1132:1188 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 8 16 theme therapeutics 1532:1543 arg1 development 1517:1527 the development 1513:1527 the development of therapeutics that target complement 1513:1566 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 1 17 theme immune 145:150 arg1 system 152:157 the innate immune system 134:157 the innate immune system 134:157 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 5 18 theme domain 1014:1019 arg1 movement 964:971 a dramatic movement 953:971 a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces 953:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 18 theme domain 1014:1019 arg1 A 978:978 >80 A 974:978 >80 A 974:978 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 1 19 theme system 152:157 arg1 part 126:129 a key part 120:129 a key part of the innate immune system 120:157 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 1 19 theme system 152:157 arg1 system 110:115 The complement system 95:115 The complement system 95:115 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 4 20 theme CRIg 756:759 arg1 mutants 761:767 CRIg mutants 756:767 CRIg mutants 756:767 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 4 21 with complex 727:733 arg1 CRIg 740:743 CRIg 740:743 CRIg 740:743 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 8 22 theme activation 1422:1431 arg1 means 1441:1445 the means 1437:1445 the means by which CRIg inhibits the convertases 1437:1484 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 8 22 theme activation 1422:1431 arg1 basis 1402:1406 the structural basis 1387:1406 the structural basis of complement activation 1387:1431 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 5 23 theme C3 899:900 arg1 activation 885:894 activation 885:894 activation of C3 885:900 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 7 24 theme macromolecular 1240:1253 arg1 rearrangements 1266:1279 the complex macromolecular structural rearrangements 1228:1279 the complex macromolecular structural rearrangements 1228:1279 The structure provides insights into the complex macromolecular structural rearrangements that occur during complement activation and inhibition. 17051150 2 25 theme third 258:262 arg1 component 264:272 the third component 254:272 the third component of the complement system 254:297 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 2 25 theme third 258:262 arg1 C3 300:301 C3 300:301 C3 300:301 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 2 26 theme factor 350:355 arg1 B 357:357 factor B 350:357 factor B 350:357 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 2 27 theme B 357:357 arg1 recruitment 335:345 recruitment 335:345 recruitment of factor B 335:357 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 3 28 theme convertase 656:665 arg1 activation 667:676 convertase activation 656:676 convertase activation 656:676 CRIg, a complement receptor expressed on macrophages, binds to C3b and iC3b mediating phagocytosis of the particles, but it is unknown how CRIg selectively recognizes proteolytic C3-fragments and whether binding of CRIg to C3b inhibits convertase activation. 17051150 8 29 theme structure-function 1351:1368 arg1 studies 1370:1376 our structure-function studies 1347:1376 our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases 1347:1484 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 2 30 theme pathway 399:405 arg1 convertases 407:417 the alternative pathway convertases 383:417 the alternative pathway convertases 383:417 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 8 31 theme important 1494:1502 arg1 clues 1504:1508 important clues 1494:1508 important clues to the development of therapeutics that target complement 1494:1566 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 4 32 theme alternative 825:835 arg1 pathway 837:843 the alternative pathway 821:843 the alternative pathway of complement 821:857 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 5 33 theme major 910:914 arg1 movement 964:971 a dramatic movement 953:971 a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces 953:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 33 theme major 910:914 arg1 rearrangements 927:940 major structural rearrangements 910:940 major structural rearrangements 910:940 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 2 34 theme alternative 387:397 arg1 convertases 407:417 the alternative pathway convertases 383:417 the alternative pathway convertases 383:417 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 1 35 from bloodstream 212:222 arg1 clearance 180:188 clearance 180:188 clearance of pathogens from the bloodstream 180:222 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 8 36 theme structural 1391:1400 arg1 basis 1402:1406 the structural basis 1387:1406 the structural basis of complement activation 1387:1431 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 5 37 theme structural 916:925 arg1 movement 964:971 a dramatic movement 953:971 a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces 953:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 37 theme structural 916:925 arg1 rearrangements 927:940 major structural rearrangements 910:940 major structural rearrangements 910:940 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 38 theme pathogen 1051:1058 arg1 surfaces 1060:1067 pathogen surfaces 1051:1067 pathogen surfaces 1051:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 39 theme thioester-bond-containing 988:1012 arg1 domain 1014:1019 the thioester-bond-containing domain 984:1019 the thioester-bond-containing domain through which C3b attaches to pathogen surfaces 984:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 1 40 theme complement 99:108 arg1 part 126:129 a key part 120:129 a key part of the innate immune system 120:157 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 1 40 theme complement 99:108 arg1 system 110:115 The complement system 95:115 The complement system 95:115 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 3 41 theme proteolytic 587:597 arg1 C3-fragments 599:610 proteolytic C3-fragments 587:610 proteolytic C3-fragments 587:610 CRIg, a complement receptor expressed on macrophages, binds to C3b and iC3b mediating phagocytosis of the particles, but it is unknown how CRIg selectively recognizes proteolytic C3-fragments and whether binding of CRIg to C3b inhibits convertase activation. 17051150 6 42 theme phagocytic 1102:1111 arg1 receptor 1113:1120 a phagocytic receptor 1100:1120 a phagocytic receptor 1100:1120 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 6 42 theme phagocytic 1102:1111 arg1 CRIg 1083:1086 CRIg 1083:1086 CRIg 1083:1086 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 2 43 theme system 292:297 arg1 component 264:272 the third component 254:272 the third component of the complement system 254:297 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 2 43 theme system 292:297 arg1 C3 300:301 C3 300:301 C3 300:301 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 4 44 theme C3b 720:722 arg1 structure 707:715 the crystal structure 695:715 the crystal structure of C3b in complex with CRIg 695:743 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 7 45 theme structural 1255:1264 arg1 rearrangements 1266:1279 the complex macromolecular structural rearrangements 1228:1279 the complex macromolecular structural rearrangements 1228:1279 The structure provides insights into the complex macromolecular structural rearrangements that occur during complement activation and inhibition. 17051150 7 46 theme complement 1299:1308 arg1 activation 1310:1319 complement activation 1299:1319 complement activation 1299:1319 The structure provides insights into the complex macromolecular structural rearrangements that occur during complement activation and inhibition. 17051150 2 47 theme complement 281:290 arg1 system 292:297 the complement system 277:297 the complement system 277:297 After exposure to pathogens, the third component of the complement system, C3, is cleaved to C3b which, after recruitment of factor B, initiates formation of the alternative pathway convertases. 17051150 6 48 theme convertases 1178:1188 arg1 receptor 1113:1120 a phagocytic receptor 1100:1120 a phagocytic receptor 1100:1120 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 6 48 theme convertases 1178:1188 arg1 inhibitor 1141:1149 a potent inhibitor 1132:1149 a potent inhibitor of the alternative pathway convertases 1132:1188 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 6 48 theme convertases 1178:1188 arg1 CRIg 1083:1086 CRIg 1083:1086 CRIg 1083:1086 We show that CRIg is not only a phagocytic receptor, but also a potent inhibitor of the alternative pathway convertases. 17051150 5 49 theme dramatic 955:962 arg1 movement 964:971 a dramatic movement 953:971 a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces 953:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 49 theme dramatic 955:962 arg1 A 978:978 >80 A 974:978 >80 A 974:978 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 50 attach attaches 1039:1046 arg1 surfaces 1060:1067 pathogen surfaces 1051:1067 pathogen surfaces 1051:1067 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 5 50 attach attaches 1039:1046 arg2 C3b 1035:1037 C3b 1035:1037 C3b 1035:1037 The structure shows that activation of C3 induces major structural rearrangements, including a dramatic movement (>80 A) of the thioester-bond-containing domain through which C3b attaches to pathogen surfaces. 17051150 8 51 theme complement 1411:1420 arg1 activation 1422:1431 complement activation 1411:1431 complement activation 1411:1431 Moreover, our structure-function studies relating the structural basis of complement activation and the means by which CRIg inhibits the convertases provide important clues to the development of therapeutics that target complement. 17051150 3 52 theme particles 526:534 arg1 phagocytosis 506:517 phagocytosis 506:517 phagocytosis of the particles 506:534 CRIg, a complement receptor expressed on macrophages, binds to C3b and iC3b mediating phagocytosis of the particles, but it is unknown how CRIg selectively recognizes proteolytic C3-fragments and whether binding of CRIg to C3b inhibits convertase activation. 17051150 1 53 theme key 122:124 arg1 part 126:129 a key part 120:129 a key part of the innate immune system 120:157 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 1 53 theme key 122:124 arg1 system 110:115 The complement system 95:115 The complement system 95:115 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 1 54 theme pathogens 193:201 arg1 clearance 180:188 clearance 180:188 clearance of pathogens from the bloodstream 180:222 The complement system is a key part of the innate immune system, and is required for clearance of pathogens from the bloodstream. 17051150 0 55 theme C3b 13:15 arg1 Structure 0:8 Structure 0:8 Structure of C3b in complex with CRIg 0:36 Structure of C3b in complex with CRIg gives insights into regulation of complement activation. 17051150 4 56 theme pathway 837:843 arg1 CRIg 792:795 CRIg 792:795 CRIg 792:795 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 17051150 4 56 theme pathway 837:843 arg1 inhibitor 808:816 an inhibitor 805:816 an inhibitor of the alternative pathway of complement 805:857 Here we present the crystal structure of C3b in complex with CRIg and, using CRIg mutants, provide evidence that CRIg acts as an inhibitor of the alternative pathway of complement. 22664871 1 0 theme multiple 294:301 arg1 roles 303:307 multiple roles 294:307 multiple roles 294:307 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 7 1 with comparison 1207:1216 arg1 complex 1246:1252 TRAIL/death receptor 5 complex 1223:1252 TRAIL/death receptor 5 complex 1223:1252 Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL. 22664871 1 2 theme signaling 132:140 arg1 RANK 151:154 its signaling receptor RANK 128:154 its signaling receptor RANK 128:154 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 3 theme molecular 215:223 arg1 triad 225:229 a molecular triad 213:229 a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system 213:328 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 3 theme molecular 215:223 arg1 critical 239:246 critical 239:246 critical 239:246 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 5 4 theme binding 844:850 arg1 determinant 852:862 the major binding determinant 834:862 the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay 834:949 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 3 5 from structure 437:445 arg1 complex 472:478 complex 472:478 complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 472:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 1 6 theme receptor 142:149 arg1 RANK 151:154 its signaling receptor RANK 128:154 its signaling receptor RANK 128:154 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 6 7 theme RANKL 1168:1172 arg1 RANKL 1168:1172 RANKL 1168:1172 RANKL 1168:1172 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 6 7 theme RANKL 1168:1172 arg1 residues 1156:1163 important interacting residues 1134:1163 important interacting residues of RANKL 1134:1172 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 5 8 theme affinity 893:900 arg1 measurement 902:912 affinity measurement 893:912 affinity measurement 893:912 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 3 9 theme N-terminal 489:498 arg1 fragment 500:507 the N-terminal fragment 485:507 the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 485:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 6 10 theme important 1134:1142 arg1 RANKL 1168:1172 RANKL 1168:1172 RANKL 1168:1172 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 6 10 theme important 1134:1142 arg1 residues 1156:1163 important interacting residues 1134:1163 important interacting residues of RANKL 1134:1172 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 5 11 theme measurement 902:912 arg1 assay 945:949 affinity measurement and osteoclast differentiation assay 893:949 affinity measurement and osteoclast differentiation assay 893:949 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 3 12 contain containing 522:531 arg2 domains 568:574 four cysteine-rich TNFR homologous domains 533:574 four cysteine-rich TNFR homologous domains (OPG-CRD) 533:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 12 contain containing 522:531 arg1 OPG 518:520 human OPG 512:520 human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 512:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 12 contain containing 522:531 arg2 OPG-CRD 577:583 OPG-CRD 577:583 OPG-CRD 577:583 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 1 13 theme immune 316:321 arg1 system 323:328 the immune system 312:328 the immune system 312:328 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 5 14 theme OPG-CRD 774:780 arg1 OPG-CRD 774:780 OPG-CRD 774:780 OPG-CRD 774:780 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 5 14 theme OPG-CRD 774:780 arg1 domain 764:769 the CRD3 domain 755:769 the CRD3 domain of OPG-CRD 755:780 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 3 15 theme trimer 462:467 arg1 structure 437:445 the 2.7-Å crystal structure 419:445 the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 419:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 6 16 theme reported 994:1001 arg1 structure 1028:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 2 17 with interaction 369:379 arg1 RANK 386:389 RANK 386:389 RANK 386:389 OPG binds RANKL directly to block its interaction with RANK. 22664871 4 18 theme OPG-CRD 711:717 arg1 monomers 719:726 three OPG-CRD monomers 705:726 three OPG-CRD monomers 705:726 The structure shows that RANKL trimer uses three equivalent grooves between two neighboring monomers to interact with three OPG-CRD monomers symmetrically. 22664871 3 19 theme 2.7-Å 423:427 arg1 structure 437:445 the 2.7-Å crystal structure 419:445 the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 419:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 5 20 theme shallow 799:805 arg1 groove 807:812 the shallow groove 795:812 the shallow groove of RANKL 795:821 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 7 21 theme TRAIL/death 1223:1233 arg1 complex 1246:1252 TRAIL/death receptor 5 complex 1223:1252 TRAIL/death receptor 5 complex 1223:1252 Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL. 22664871 1 22 theme Receptor 84:91 arg1 activator 93:101 Receptor activator 84:101 Receptor activator of NF-κB ligand (RANKL) 84:125 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 5 23 from domain 764:769 arg1 loop 745:748 A loop 743:748 A loop from the CRD3 domain of OPG-CRD 743:780 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 6 24 theme complex 1020:1026 arg1 structure 1028:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 0 25 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human RANKL 0:31 Crystal structure of human RANKL complexed with its decoy receptor osteoprotegerin. 22664871 4 26 theme neighboring 667:677 arg1 monomers 679:686 two neighboring monomers 663:686 two neighboring monomers 663:686 The structure shows that RANKL trimer uses three equivalent grooves between two neighboring monomers to interact with three OPG-CRD monomers symmetrically. 22664871 6 27 theme RANKL/RANK 1009:1018 arg1 structure 1028:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 0 28 theme human 21:25 arg1 RANKL 27:31 human RANKL 21:31 human RANKL 21:31 Crystal structure of human RANKL complexed with its decoy receptor osteoprotegerin. 22664871 6 29 theme RANK 1178:1181 arg1 recognition 1183:1193 RANK recognition 1178:1193 RANK recognition 1178:1193 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 1 30 theme decoy 165:169 arg1 OPG 197:199 OPG 197:199 OPG 197:199 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 30 theme decoy 165:169 arg1 osteoprotegerin 180:194 its decoy receptor osteoprotegerin 161:194 its decoy receptor osteoprotegerin (OPG) 161:200 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 31 theme NF-κB 106:110 arg1 RANKL 120:124 RANKL 120:124 RANKL 120:124 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 31 theme NF-κB 106:110 arg1 ligand 112:117 NF-κB ligand 106:117 NF-κB ligand (RANKL) 106:125 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 32 theme receptor 171:178 arg1 OPG 197:199 OPG 197:199 OPG 197:199 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 32 theme receptor 171:178 arg1 osteoprotegerin 180:194 its decoy receptor osteoprotegerin 161:194 its decoy receptor osteoprotegerin (OPG) 161:200 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 33 theme bone 262:265 arg1 remodeling 267:276 bone remodeling 262:276 bone remodeling 262:276 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 6 34 theme receptor 1072:1079 arg1 function 1081:1088 its decoy receptor function 1062:1088 its decoy receptor function 1062:1088 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 7 35 theme structural 1267:1276 arg1 basis 1278:1282 structural basis 1267:1282 structural basis for the cross-reactivity of OPG to TRAIL 1267:1323 Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL. 22664871 0 36 theme RANKL 27:31 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human RANKL 0:31 Crystal structure of human RANKL complexed with its decoy receptor osteoprotegerin. 22664871 7 37 theme OPG 1312:1314 arg1 cross-reactivity 1292:1307 the cross-reactivity 1288:1307 the cross-reactivity of OPG to TRAIL 1288:1323 Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL. 22664871 5 38 theme CRD3 759:762 arg1 OPG-CRD 774:780 OPG-CRD 774:780 OPG-CRD 774:780 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 5 38 theme CRD3 759:762 arg1 domain 764:769 the CRD3 domain 755:769 the CRD3 domain of OPG-CRD 755:780 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 3 39 with complex 472:478 arg1 fragment 500:507 the N-terminal fragment 485:507 the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 485:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 5 40 theme RANKL 817:821 arg1 groove 807:812 the shallow groove 795:812 the shallow groove of RANKL 795:821 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 7 41 theme Structural 1196:1205 arg1 comparison 1207:1216 Structural comparison 1196:1216 Structural comparison with TRAIL/death receptor 5 complex 1196:1252 Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL. 22664871 3 42 theme human 512:516 arg1 OPG 518:520 human OPG 512:520 human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 512:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 1 43 theme ligand 112:117 arg1 activator 93:101 Receptor activator 84:101 Receptor activator of NF-κB ligand (RANKL) 84:125 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 43 theme ligand 112:117 arg1 RANK 151:154 its signaling receptor RANK 128:154 its signaling receptor RANK 128:154 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 43 theme ligand 112:117 arg1 osteoprotegerin 180:194 its decoy receptor osteoprotegerin 161:194 its decoy receptor osteoprotegerin (OPG) 161:200 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 1 43 theme ligand 112:117 arg1 OPG 197:199 OPG 197:199 OPG 197:199 Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. 22664871 3 44 theme OPG 518:520 arg1 fragment 500:507 the N-terminal fragment 485:507 the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 485:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 45 theme crystal 429:435 arg1 structure 437:445 the 2.7-Å crystal structure 419:445 the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD) 419:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 6 46 theme residues 1156:1163 arg1 accessibilities 1115:1129 the accessibilities 1111:1129 the accessibilities of important interacting residues of RANKL for RANK recognition 1111:1193 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 5 47 theme differentiation 929:943 arg1 assay 945:949 affinity measurement and osteoclast differentiation assay 893:949 affinity measurement and osteoclast differentiation assay 893:949 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 3 48 theme homologous 557:566 arg1 OPG-CRD 577:583 OPG-CRD 577:583 OPG-CRD 577:583 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 48 theme homologous 557:566 arg1 domains 568:574 four cysteine-rich TNFR homologous domains 533:574 four cysteine-rich TNFR homologous domains (OPG-CRD) 533:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 7 49 theme receptor 1235:1242 arg1 complex 1246:1252 TRAIL/death receptor 5 complex 1223:1252 TRAIL/death receptor 5 complex 1223:1252 Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL. 22664871 6 50 theme decoy 1066:1070 arg1 function 1081:1088 its decoy receptor function 1062:1088 its decoy receptor function 1062:1088 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 5 51 theme osteoclast 918:927 arg1 differentiation 929:943 osteoclast differentiation 918:943 osteoclast differentiation 918:943 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 5 52 theme major 838:842 arg1 determinant 852:862 the major binding determinant 834:862 the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay 834:949 A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. 22664871 6 53 theme interacting 1144:1154 arg1 RANKL 1168:1172 RANKL 1168:1172 RANKL 1168:1172 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 6 53 theme interacting 1144:1154 arg1 residues 1156:1163 important interacting residues 1134:1163 important interacting residues of RANKL 1134:1172 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 4 54 theme RANKL 612:616 arg1 trimer 618:623 RANKL trimer 612:623 RANKL trimer 612:623 The structure shows that RANKL trimer uses three equivalent grooves between two neighboring monomers to interact with three OPG-CRD monomers symmetrically. 22664871 0 55 theme receptor 58:65 arg1 osteoprotegerin 67:81 its decoy receptor osteoprotegerin 48:81 its decoy receptor osteoprotegerin 48:81 Crystal structure of human RANKL complexed with its decoy receptor osteoprotegerin. 22664871 3 56 theme cysteine-rich 538:550 arg1 OPG-CRD 577:583 OPG-CRD 577:583 OPG-CRD 577:583 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 56 theme cysteine-rich 538:550 arg1 domains 568:574 four cysteine-rich TNFR homologous domains 533:574 four cysteine-rich TNFR homologous domains (OPG-CRD) 533:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 6 57 theme mouse 1003:1007 arg1 structure 1028:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 a previously reported mouse RANKL/RANK complex structure 981:1036 These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. 22664871 3 58 theme human 450:454 arg1 trimer 462:467 human RANKL trimer 450:467 human RANKL trimer 450:467 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 0 59 theme decoy 52:56 arg1 osteoprotegerin 67:81 its decoy receptor osteoprotegerin 48:81 its decoy receptor osteoprotegerin 48:81 Crystal structure of human RANKL complexed with its decoy receptor osteoprotegerin. 22664871 3 60 theme TNFR 552:555 arg1 OPG-CRD 577:583 OPG-CRD 577:583 OPG-CRD 577:583 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 60 theme TNFR 552:555 arg1 domains 568:574 four cysteine-rich TNFR homologous domains 533:574 four cysteine-rich TNFR homologous domains (OPG-CRD) 533:584 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 3 61 theme RANKL 456:460 arg1 trimer 462:467 human RANKL trimer 450:467 human RANKL trimer 450:467 In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). 22664871 4 62 theme equivalent 636:645 arg1 grooves 647:653 three equivalent grooves 630:653 three equivalent grooves between two neighboring monomers 630:686 The structure shows that RANKL trimer uses three equivalent grooves between two neighboring monomers to interact with three OPG-CRD monomers symmetrically. 1569071 7 0 theme apparent 1486:1493 arg1 Mr 1495:1496 apparent Mr 1486:1496 apparent Mr = 15,000 and 11,500 IGF-II 1486:1523 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 10 1 theme mobility 2041:2048 arg1 changes 2050:2056 relative mobility changes 2032:2056 relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors 2032:2173 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 3 2 theme acid 652:655 arg1 extracts 657:664 the acetic acid extracts 641:664 the acetic acid extracts 641:664 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II was also identified in the acetic acid extracts. 1569071 6 3 theme endoprotease-generated 1224:1245 arg1 peptides 1247:1254 their endoprotease-generated peptides 1218:1254 their endoprotease-generated peptides 1218:1254 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 1 4 theme IV1 443:445 arg1 extracts 417:424 acetic acid extracts 405:424 acetic acid extracts of Cohn fraction IV1 405:445 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 5 theme acid 412:415 arg1 extracts 417:424 acetic acid extracts 405:424 acetic acid extracts of Cohn fraction IV1 405:445 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 6 6 theme gel 1438:1440 arg1 electrophoresis 1442:1456 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1400:1456 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1400:1456 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 1 7 theme pressure 268:275 arg1 chromatography 284:297 reversed-phase high pressure liquid chromatography 248:297 reversed-phase high pressure liquid chromatography 248:297 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 11 8 theme binding 2315:2321 arg1 specificity 2300:2310 the specificity 2296:2310 the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin 2296:2379 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 11 9 link O-linked 2204:2211 arg1 oligosaccharide 2213:2227 O-linked oligosaccharide 2204:2227 O-linked oligosaccharide 2204:2227 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 11 10 gly asialoglycopeptide 2342:2359 arg2 asialoglycopeptide 2342:2359 the Ser66-Lys88 asialoglycopeptide 2326:2359 the Ser66-Lys88 asialoglycopeptide 2326:2359 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 6 11 from change 1331:1336 arg1 peptides 1384:1391 peptides 1384:1391 peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1384:1456 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 6 11 from change 1331:1336 arg1 mobilities 1350:1359 relative mobilities 1341:1359 relative mobilities of the polypeptide 1341:1378 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 2 12 contain had 463:465 arg2 pI 487:488 pI 487:488 pI 487:488 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 12 contain had 463:465 arg1 precursors 452:461 Two precursors 448:461 Two precursors 448:461 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 12 contain had 463:465 arg2 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 12 contain had 463:465 arg2 points 479:484 isoelectric points 467:484 isoelectric points (pI) of 5.1 and 5.4 467:504 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 11 13 theme E 2238:2238 arg1 peptide 2247:2253 the E domain peptide 2234:2253 the E domain peptide of IGF-II 2234:2263 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 10 14 theme E 1935:1935 arg1 peptides 1944:1951 the Ser66-Lys88 endoprotease-generated E domain peptides 1896:1951 the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II 1896:1985 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 6 15 theme high 1184:1187 arg1 weight 1199:1204 the 125I-labeled high molecular weight 1167:1204 the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides 1167:1254 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 9 16 gly unglycosylated 1800:1813 arg1 form 1815:1818 an unglycosylated form 1797:1818 an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88 1797:1888 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 9 16 gly unglycosylated 1800:1813 arg1 IGF-II 1833:1838 precursor IGF-II 1823:1838 precursor IGF-II 1823:1838 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 2 17 theme 15,000 532:537 arg1 pI 487:488 pI 487:488 pI 487:488 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 17 theme 15,000 532:537 arg1 points 479:484 isoelectric points 467:484 isoelectric points (pI) of 5.1 and 5.4 467:504 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 17 theme 15,000 532:537 arg1 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 6 18 theme 125I-labeled 1171:1182 arg1 weight 1199:1204 the 125I-labeled high molecular weight 1167:1204 the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides 1167:1254 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 10 19 theme gel 2103:2105 arg1 electrophoresis 2107:2121 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 2065:2121 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 2065:2121 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 7 20 dep = 1498:1498 arg1 IGF-II 1518:1523 15,000 and 11,500 IGF-II 1500:1523 15,000 and 11,500 IGF-II 1500:1523 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 5 21 theme amino 1014:1018 arg1 acid 1020:1023 derivatized amino acid 1002:1023 derivatized amino acid 1002:1023 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 4 22 dep = 785:785 arg1 pI 802:803 pI 5.1 802:807 pI 5.1 802:807 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 22 dep = 785:785 arg1 IGF-II 794:799 15,000 IGF-II 787:799 15,000 IGF-II (pI 5.1) 787:808 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 23 theme endoprotease 844:855 arg1 Lys-C 857:861 the endoprotease Lys-C 840:861 the endoprotease Lys-C 840:861 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 24 theme acid 688:691 arg1 sequencing 693:702 Amino-terminal amino acid sequencing 667:702 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 667:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 2 25 theme 11,500 543:548 arg1 pI 487:488 pI 487:488 pI 487:488 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 25 theme 11,500 543:548 arg1 points 479:484 isoelectric points 467:484 isoelectric points (pI) of 5.1 and 5.4 467:504 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 25 theme 11,500 543:548 arg1 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 10 26 theme Mr 1977:1978 arg1 IGF-II 1980:1985 pI5.1 and 5.4 high Mr IGF-II 1958:1985 pI5.1 and 5.4 high Mr IGF-II 1958:1985 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 5 27 theme reduced 985:991 arg1 yield 993:997 a sharply reduced yield 975:997 a sharply reduced yield of derivatized amino acid 975:1023 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 7 28 theme 10,500 1570:1575 arg1 value 1561:1565 a common value 1552:1565 a common value of 10,500 1552:1575 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 2 29 theme 5.1 494:496 arg1 pI 487:488 pI 487:488 pI 487:488 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 29 theme 5.1 494:496 arg1 points 479:484 isoelectric points 467:484 isoelectric points (pI) of 5.1 and 5.4 467:504 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 29 theme 5.1 494:496 arg1 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 6 30 theme relative 1341:1348 arg1 mobilities 1350:1359 relative mobilities 1341:1359 relative mobilities of the polypeptide 1341:1378 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 3 31 theme 16,000 582:587 arg1 variant 600:606 An apparent Mr = 16,000 RLPG/Ser29 variant 565:606 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II 565:616 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II was also identified in the acetic acid extracts. 1569071 4 32 theme Amino-terminal 667:680 arg1 sequencing 693:702 Amino-terminal amino acid sequencing 667:702 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 667:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 33 attach isolated 759:766 arg1 Mr 782:783 apparent Mr 773:783 apparent Mr = 15,000 IGF-II (pI 5.1) 773:808 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 33 attach isolated 759:766 arg2 peptide 737:743 the major E domain-containing peptide 707:743 the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 707:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 1 34 theme insulin-like 361:372 arg1 factor 381:386 human insulin-like growth factor II 355:389 human insulin-like growth factor II (IGF-II) 355:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 34 theme insulin-like 361:372 arg1 IGF-II 392:397 IGF-II 392:397 IGF-II 392:397 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 35 theme Cohn 429:432 arg1 IV1 443:445 Cohn fraction IV1 429:445 Cohn fraction IV1 429:445 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 36 theme growth 155:160 arg1 protein 177:183 insulin-like growth factor-binding protein 3 142:185 insulin-like growth factor-binding protein 3 (IGF-BP-3) 142:196 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 36 theme growth 155:160 arg1 IGF-BP-3 188:195 IGF-BP-3 188:195 IGF-BP-3 188:195 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 2 37 theme apparent 510:517 arg1 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 11 38 theme jackfruit 2364:2372 arg1 lectin 2374:2379 jackfruit lectin 2364:2379 jackfruit lectin 2364:2379 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 1 39 theme factor 381:386 arg1 precursors 341:350 high molecular weight precursors 319:350 high molecular weight precursors of human insulin-like growth factor II (IGF-II) 319:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 2 40 theme 5.4 502:504 arg1 pI 487:488 pI 487:488 pI 487:488 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 40 theme 5.4 502:504 arg1 points 479:484 isoelectric points 467:484 isoelectric points (pI) of 5.1 and 5.4 467:504 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 40 theme 5.4 502:504 arg1 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 10 41 theme sulfate-polyacrylamide 2080:2101 arg1 electrophoresis 2107:2121 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 2065:2121 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 2065:2121 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 11 42 theme asialoglycopeptide 2342:2359 arg1 binding 2315:2321 binding 2315:2321 binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin 2315:2379 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 1 43 dep exchange 112:119 arg1 chromatography 207:220 affinity chromatography 198:220 affinity chromatography 198:220 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 9 44 theme Mr 1760:1761 arg1 This 1747:1750 This 1747:1750 This 1747:1750 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 9 44 theme Mr 1760:1761 arg1 value 1763:1767 the Mr value 1756:1767 the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88 1756:1888 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 4 45 theme major 711:715 arg1 peptide 737:743 the major E domain-containing peptide 707:743 the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 707:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 11 46 theme domain 2240:2245 arg1 peptide 2247:2253 the E domain peptide 2234:2253 the E domain peptide of IGF-II 2234:2263 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 4 47 theme domain-containing 719:735 arg1 peptide 737:743 the major E domain-containing peptide 707:743 the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 707:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 1 48 theme ion 108:110 arg1 exchange 112:119 ion exchange 108:119 ion exchange 108:119 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 0 49 theme factor 71:76 arg1 precursors 37:46 O-glycosylated precursors 22:46 O-glycosylated precursors of insulin-like growth factor II 22:79 The identification of O-glycosylated precursors of insulin-like growth factor II. 1569071 6 50 theme polypeptide 1368:1378 arg1 peptides 1384:1391 peptides 1384:1391 peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1384:1456 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 6 50 theme polypeptide 1368:1378 arg1 mobilities 1350:1359 relative mobilities 1341:1359 relative mobilities of the polypeptide 1341:1378 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 1 51 theme high 319:322 arg1 precursors 341:350 high molecular weight precursors 319:350 high molecular weight precursors of human insulin-like growth factor II (IGF-II) 319:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 0 52 theme precursors 37:46 arg1 identification 4:17 The identification 0:17 The identification of O-glycosylated precursors of insulin-like growth factor II. 0:80 The identification of O-glycosylated precursors of insulin-like growth factor II. 1569071 2 53 theme isoelectric 467:477 arg1 pI 487:488 pI 487:488 pI 487:488 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 2 53 theme isoelectric 467:477 arg1 points 479:484 isoelectric points 467:484 isoelectric points (pI) of 5.1 and 5.4 467:504 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 3 54 theme IGF-II 611:616 arg1 variant 600:606 An apparent Mr = 16,000 RLPG/Ser29 variant 565:606 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II 565:616 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II was also identified in the acetic acid extracts. 1569071 0 55 theme insulin-like 51:62 arg1 factor 71:76 insulin-like growth factor II 51:79 insulin-like growth factor II 51:79 The identification of O-glycosylated precursors of insulin-like growth factor II. 1569071 1 56 theme weight 334:339 arg1 precursors 341:350 high molecular weight precursors 319:350 high molecular weight precursors of human insulin-like growth factor II (IGF-II) 319:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 10 57 from IGF-II 1980:1985 arg1 peptides 1944:1951 the Ser66-Lys88 endoprotease-generated E domain peptides 1896:1951 the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II 1896:1985 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 0 58 gly O-glycosylated 22:35 arg1 precursors 37:46 O-glycosylated precursors 22:46 O-glycosylated precursors of insulin-like growth factor II 22:79 The identification of O-glycosylated precursors of insulin-like growth factor II. 1569071 9 59 theme unglycosylated 1800:1813 arg1 form 1815:1818 an unglycosylated form 1797:1818 an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88 1797:1888 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 3 60 theme acetic 645:650 arg1 extracts 657:664 the acetic acid extracts 641:664 the acetic acid extracts 641:664 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II was also identified in the acetic acid extracts. 1569071 7 61 theme Mr 1495:1496 arg1 treatment 1473:1481 Neuraminidase treatment 1459:1481 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II 1459:1523 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 11 62 theme IGF-II 2258:2263 arg1 peptide 2247:2253 the E domain peptide 2234:2253 the E domain peptide of IGF-II 2234:2263 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 9 63 theme IGF-II 1833:1838 arg1 form 1815:1818 an unglycosylated form 1797:1818 an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88 1797:1888 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 1 64 theme high 263:266 arg1 chromatography 284:297 reversed-phase high pressure liquid chromatography 248:297 reversed-phase high pressure liquid chromatography 248:297 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 6 65 dep weight 1199:1204 arg1 IGF-IIs 1206:1212 IGF-IIs 1206:1212 IGF-IIs 1206:1212 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 6 65 dep weight 1199:1204 arg1 peptides 1247:1254 their endoprotease-generated peptides 1218:1254 their endoprotease-generated peptides 1218:1254 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 7 66 theme Neuraminidase 1459:1471 arg1 treatment 1473:1481 Neuraminidase treatment 1459:1481 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II 1459:1523 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 10 67 theme relative 2032:2039 arg1 changes 2050:2056 relative mobility changes 2032:2056 relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors 2032:2173 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 6 68 theme sulfate-polyacrylamide 1415:1436 arg1 electrophoresis 1442:1456 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1400:1456 sodium dodecyl sulfate-polyacrylamide gel electrophoresis 1400:1456 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 1 69 theme liquid 277:282 arg1 chromatography 284:297 reversed-phase high pressure liquid chromatography 248:297 reversed-phase high pressure liquid chromatography 248:297 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 9 70 theme carboxyl-terminal 1851:1867 arg1 end 1869:1871 a carboxyl-terminal end 1849:1871 a carboxyl-terminal end 1849:1871 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 8 71 theme IGF-II 1614:1619 arg1 precursors 1600:1609 the desialylated precursors 1583:1609 the desialylated precursors of IGF-II 1583:1619 When the desialylated precursors of IGF-II were treated with O-glycosidase, but not when treated with N-glycosidase, the Mr values were reduced further to about 10,000. 1569071 1 72 theme factor-binding 162:175 arg1 protein 177:183 insulin-like growth factor-binding protein 3 142:185 insulin-like growth factor-binding protein 3 (IGF-BP-3) 142:196 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 72 theme factor-binding 162:175 arg1 IGF-BP-3 188:195 IGF-BP-3 188:195 IGF-BP-3 188:195 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 73 used used 303:306 arg2 procedure 84:92 A procedure 82:92 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography 82:297 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 6 74 theme molecular 1189:1197 arg1 weight 1199:1204 the 125I-labeled high molecular weight 1167:1204 the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides 1167:1254 To evaluate this possibility, the 125I-labeled high molecular weight IGF-IIs and their endoprotease-generated peptides were treated with glycosidases, and their effects were determined from the change in relative mobilities of the polypeptide and peptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 1569071 10 75 theme endoprotease-generated 1912:1933 arg1 peptides 1944:1951 the Ser66-Lys88 endoprotease-generated E domain peptides 1896:1951 the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II 1896:1985 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 8 76 gly desialylated 1587:1598 arg1 precursors 1600:1609 the desialylated precursors 1583:1609 the desialylated precursors of IGF-II 1583:1619 When the desialylated precursors of IGF-II were treated with O-glycosidase, but not when treated with N-glycosidase, the Mr values were reduced further to about 10,000. 1569071 8 77 theme Mr 1699:1700 arg1 values 1702:1707 the Mr values 1695:1707 the Mr values 1695:1707 When the desialylated precursors of IGF-II were treated with O-glycosidase, but not when treated with N-glycosidase, the Mr values were reduced further to about 10,000. 1569071 5 78 theme derivatized 1002:1012 arg1 acid 1020:1023 derivatized amino acid 1002:1023 derivatized amino acid 1002:1023 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 1 79 theme affinity 198:205 arg1 chromatography 207:220 affinity chromatography 198:220 affinity chromatography 198:220 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 5 80 mod modified 1098:1105 arg3 O-glycosylation 1120:1134 O-glycosylation 1120:1134 O-glycosylation 1120:1134 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 5 80 mod modified 1098:1105 arg1 Thr75 1063:1067 Thr75 1063:1067 Thr75 1063:1067 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 4 81 theme carboxyl 878:885 arg1 terminus 887:894 the carboxyl terminus 874:894 the carboxyl terminus of this precursor 874:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 11 82 theme oligosaccharide 2213:2227 arg1 association 2189:2199 the association 2185:2199 the association of O-linked oligosaccharide with the E domain peptide of IGF-II 2185:2263 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 5 83 theme acid 1020:1023 arg1 yield 993:997 a sharply reduced yield 975:997 a sharply reduced yield of derivatized amino acid 975:1023 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 9 84 contain had 1845:1847 arg1 form 1815:1818 an unglycosylated form 1797:1818 an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88 1797:1888 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 9 84 contain had 1845:1847 arg2 end 1869:1871 a carboxyl-terminal end 1849:1871 a carboxyl-terminal end 1849:1871 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 4 85 theme amino 682:686 arg1 sequencing 693:702 Amino-terminal amino acid sequencing 667:702 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 667:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 10 86 theme domain 1937:1942 arg1 peptides 1944:1951 the Ser66-Lys88 endoprotease-generated E domain peptides 1896:1951 the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II 1896:1985 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 3 87 theme RLPG/Ser29 589:598 arg1 variant 600:606 An apparent Mr = 16,000 RLPG/Ser29 variant 565:606 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II 565:616 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II was also identified in the acetic acid extracts. 1569071 1 88 theme gel 122:124 arg1 permeation 126:135 gel permeation 122:135 gel permeation 122:135 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 4 89 with digestion 825:833 arg1 Lys-C 857:861 the endoprotease Lys-C 840:861 the endoprotease Lys-C 840:861 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 10 90 theme high 1972:1975 arg1 IGF-II 1980:1985 pI5.1 and 5.4 high Mr IGF-II 1958:1985 pI5.1 and 5.4 high Mr IGF-II 1958:1985 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 1 91 theme fraction 434:441 arg1 IV1 443:445 Cohn fraction IV1 429:445 Cohn fraction IV1 429:445 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 3 92 theme = 580:580 arg1 variant 600:606 An apparent Mr = 16,000 RLPG/Ser29 variant 565:606 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II 565:616 An apparent Mr = 16,000 RLPG/Ser29 variant of IGF-II was also identified in the acetic acid extracts. 1569071 7 93 theme common 1554:1559 arg1 value 1561:1565 a common value 1552:1565 a common value of 10,500 1552:1575 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 11 94 with association 2189:2199 arg1 peptide 2247:2253 the E domain peptide 2234:2253 the E domain peptide of IGF-II 2234:2263 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 10 95 theme intact 2157:2162 arg1 precursors 2164:2173 the intact precursors 2153:2173 the intact precursors 2153:2173 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 1 96 theme human 355:359 arg1 factor 381:386 human insulin-like growth factor II 355:389 human insulin-like growth factor II (IGF-II) 355:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 96 theme human 355:359 arg1 IGF-II 392:397 IGF-II 392:397 IGF-II 392:397 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 97 theme insulin-like 142:153 arg1 protein 177:183 insulin-like growth factor-binding protein 3 142:185 insulin-like growth factor-binding protein 3 (IGF-BP-3) 142:196 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 97 theme insulin-like 142:153 arg1 IGF-BP-3 188:195 IGF-BP-3 188:195 IGF-BP-3 188:195 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 98 theme growth 374:379 arg1 factor 381:386 human insulin-like growth factor II 355:389 human insulin-like growth factor II (IGF-II) 355:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 1 98 theme growth 374:379 arg1 IGF-II 392:397 IGF-II 392:397 IGF-II 392:397 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 7 99 theme Mr 1539:1540 arg1 values 1542:1547 their Mr values 1533:1547 their Mr values 1533:1547 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 11 100 theme Ser66-Lys88 2330:2340 arg1 asialoglycopeptide 2342:2359 the Ser66-Lys88 asialoglycopeptide 2326:2359 the Ser66-Lys88 asialoglycopeptide 2326:2359 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 4 101 theme peptide 737:743 arg1 sequencing 693:702 Amino-terminal amino acid sequencing 667:702 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 667:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 102 theme precursor 904:912 arg1 terminus 887:894 the carboxyl terminus 874:894 the carboxyl terminus of this precursor 874:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 0 103 theme O-glycosylated 22:35 arg1 precursors 37:46 O-glycosylated precursors 22:46 O-glycosylated precursors of insulin-like growth factor II 22:79 The identification of O-glycosylated precursors of insulin-like growth factor II. 1569071 4 104 theme E 717:717 arg1 peptide 737:743 the major E domain-containing peptide 707:743 the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor 707:912 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 4 105 theme = 785:785 arg1 Mr 782:783 apparent Mr 773:783 apparent Mr = 15,000 IGF-II (pI 5.1) 773:808 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 11 106 theme O-linked 2204:2211 arg1 oligosaccharide 2213:2227 O-linked oligosaccharide 2204:2227 O-linked oligosaccharide 2204:2227 Finally, the association of O-linked oligosaccharide with the E domain peptide of IGF-II was confirmed by demonstrating the specificity of binding of the Ser66-Lys88 asialoglycopeptide to jackfruit lectin. 1569071 0 107 theme growth 64:69 arg1 factor 71:76 insulin-like growth factor II 51:79 insulin-like growth factor II 51:79 The identification of O-glycosylated precursors of insulin-like growth factor II. 1569071 1 108 theme molecular 324:332 arg1 precursors 341:350 high molecular weight precursors 319:350 high molecular weight precursors of human insulin-like growth factor II (IGF-II) 319:398 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 5 109 theme peptide 966:972 arg1 sequencing 947:956 the sequencing 943:956 the sequencing of this peptide 943:972 During the sequencing of this peptide, a sharply reduced yield of derivatized amino acid occurred at cycle 10, indicating that Thr75 had been posttranslationally modified, possibly by O-glycosylation. 1569071 2 110 theme Mr 519:520 arg1 values 522:527 apparent Mr values 510:527 apparent Mr values of 15,000 and 11,500 510:548 Two precursors had isoelectric points (pI) of 5.1 and 5.4 and apparent Mr values of 15,000 and 11,500, respectively. 1569071 10 111 contain had 2028:2030 arg2 changes 2050:2056 relative mobility changes 2032:2056 relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors 2032:2173 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 10 111 contain had 2028:2030 arg1 they 2023:2026 they 2023:2026 they 2023:2026 When the Ser66-Lys88 endoprotease-generated E domain peptides from pI5.1 and 5.4 high Mr IGF-II were treated with the glycosidases, they had relative mobility changes during sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were similar to those of the intact precursors. 1569071 1 112 theme acetic 405:410 arg1 extracts 417:424 acetic acid extracts 405:424 acetic acid extracts of Cohn fraction IV1 405:445 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 1569071 7 113 theme = 1498:1498 arg1 Mr 1495:1496 apparent Mr 1486:1496 apparent Mr = 15,000 and 11,500 IGF-II 1486:1523 Neuraminidase treatment of apparent Mr = 15,000 and 11,500 IGF-II reduced their Mr values to a common value of 10,500. 1569071 9 114 theme precursor 1823:1831 arg1 IGF-II 1833:1838 precursor IGF-II 1823:1838 precursor IGF-II 1823:1838 This was the Mr value that would be predicted for an unglycosylated form of precursor IGF-II that had a carboxyl-terminal end at or near Lys88. 1569071 8 115 theme desialylated 1587:1598 arg1 precursors 1600:1609 the desialylated precursors 1583:1609 the desialylated precursors of IGF-II 1583:1619 When the desialylated precursors of IGF-II were treated with O-glycosidase, but not when treated with N-glycosidase, the Mr values were reduced further to about 10,000. 1569071 4 116 theme apparent 773:780 arg1 Mr 782:783 apparent Mr 773:783 apparent Mr = 15,000 IGF-II (pI 5.1) 773:808 Amino-terminal amino acid sequencing of the major E domain-containing peptide that had been isolated from apparent Mr = 15,000 IGF-II (pI 5.1), following its digestion with the endoprotease Lys-C, indicated the carboxyl terminus of this precursor was near or at Lys88. 1569071 1 117 theme reversed-phase 248:261 arg1 chromatography 284:297 reversed-phase high pressure liquid chromatography 248:297 reversed-phase high pressure liquid chromatography 248:297 A procedure that combined ion exchange, gel permeation, and insulin-like growth factor-binding protein 3 (IGF-BP-3) affinity chromatography with chromatofocusing and reversed-phase high pressure liquid chromatography was used to isolate high molecular weight precursors of human insulin-like growth factor II (IGF-II) from acetic acid extracts of Cohn fraction IV1. 22483115 5 0 theme conserved 756:764 arg1 patch 774:778 A conserved surface patch 754:778 A conserved surface patch in the DS domain outside the collagen-binding site 754:829 A conserved surface patch in the DS domain outside the collagen-binding site is shown to be required for signaling. 22483115 7 1 theme extracellular 1117:1129 arg1 association 1131:1141 the extracellular association 1113:1141 the extracellular association of DDR1 subunits 1113:1158 The mAbs likely inhibit signaling by sterically blocking the extracellular association of DDR1 subunits. 22483115 1 2 theme discoidin 148:156 arg1 DDR1 176:179 DDR1 176:179 DDR1 176:179 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 2 theme discoidin 148:156 arg1 receptors 165:173 The discoidin domain receptors 144:173 The discoidin domain receptors 144:173 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 2 theme discoidin 148:156 arg1 kinases 236:242 constitutively dimeric receptor tyrosine kinases 195:242 constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen 195:288 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 2 theme discoidin 148:156 arg1 DDR2 185:188 DDR2 185:188 DDR2 185:188 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 4 3 theme mAb 592:594 arg1 3E3 596:598 mAb 3E3 592:598 mAb 3E3 592:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 5 4 theme surface 766:772 arg1 patch 774:778 A conserved surface patch 754:778 A conserved surface patch in the DS domain outside the collagen-binding site 754:829 A conserved surface patch in the DS domain outside the collagen-binding site is shown to be required for signaling. 22483115 4 5 theme monomeric 527:535 arg1 region 556:561 the monomeric DDR1 extracellular region 523:561 the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 523:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 1 6 theme domain 158:163 arg1 DDR1 176:179 DDR1 176:179 DDR1 176:179 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 6 theme domain 158:163 arg1 receptors 165:173 The discoidin domain receptors 144:173 The discoidin domain receptors 144:173 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 6 theme domain 158:163 arg1 kinases 236:242 constitutively dimeric receptor tyrosine kinases 195:242 constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen 195:288 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 6 theme domain 158:163 arg1 DDR2 185:188 DDR2 185:188 DDR2 185:188 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 0 7 theme Fab 89:91 arg1 fragment 93:100 an inhibitory Fab fragment 75:100 an inhibitory Fab fragment 75:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 6 8 theme active 880:885 arg1 conformation 887:898 the active conformation 876:898 the active conformation of the DDR1 dimer 876:916 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 2 9 theme Aberrant 291:298 arg1 signaling 304:312 Aberrant DDR signaling 291:312 Aberrant DDR signaling 291:312 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 0 10 theme inhibitory 78:87 arg1 fragment 93:100 an inhibitory Fab fragment 75:100 an inhibitory Fab fragment 75:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 4 11 contain contains 719:726 arg2 epitopes 732:739 the epitopes 728:739 the epitopes of all mAbs 728:751 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 4 11 contain contains 719:726 arg1 domain 705:710 the following DS-like domain 683:710 the following DS-like domain 683:710 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 6 12 dep association 1018:1028 arg1 addition 980:987 addition 980:987 addition 980:987 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 2 13 theme many 366:369 arg1 cancers 371:377 many cancers 366:377 many cancers 366:377 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 4 14 theme 3E3 596:598 arg1 fragment 580:587 the Fab fragment 572:587 the Fab fragment of mAb 3E3 572:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 4 15 theme DS-like 697:703 arg1 domain 705:710 the following DS-like domain 683:710 the following DS-like domain 683:710 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 6 16 theme transmembrane 1033:1045 arg1 helices 1047:1053 transmembrane helices 1033:1053 transmembrane helices 1033:1053 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 3 17 theme DDR1 440:443 arg1 signaling 445:453 DDR1 signaling 440:453 DDR1 signaling 440:453 We have generated monoclonal antibodies (mAbs) that inhibit DDR1 signaling without interfering with collagen binding. 22483115 7 18 theme DDR1 1146:1149 arg1 subunits 1151:1158 DDR1 subunits 1146:1158 DDR1 subunits 1146:1158 The mAbs likely inhibit signaling by sterically blocking the extracellular association of DDR1 subunits. 22483115 4 19 theme crystal 502:508 arg1 structure 510:518 The crystal structure 498:518 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 498:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 7 20 theme subunits 1151:1158 arg1 association 1131:1141 the extracellular association 1113:1141 the extracellular association of DDR1 subunits 1113:1158 The mAbs likely inhibit signaling by sterically blocking the extracellular association of DDR1 subunits. 22483115 6 21 theme DS 965:966 arg1 domains 968:974 the DS domains 961:974 the DS domains 961:974 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 3 22 theme monoclonal 398:407 arg1 mAbs 421:424 mAbs 421:424 mAbs 421:424 We have generated monoclonal antibodies (mAbs) that inhibit DDR1 signaling without interfering with collagen binding. 22483115 3 22 theme monoclonal 398:407 arg1 antibodies 409:418 monoclonal antibodies 398:418 monoclonal antibodies (mAbs) that inhibit DDR1 signaling without interfering with collagen binding 398:495 We have generated monoclonal antibodies (mAbs) that inhibit DDR1 signaling without interfering with collagen binding. 22483115 1 23 theme triple-helical 266:279 arg1 collagen 281:288 triple-helical collagen 266:288 triple-helical collagen 266:288 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 0 24 theme discoidin 17:25 arg1 receptor 34:41 the discoidin domain receptor 1 13:43 the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment 13:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 0 25 theme important 119:127 arg1 features 110:117 features 110:117 features important for signaling 110:141 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 4 26 theme mAbs 748:751 arg1 epitopes 732:739 the epitopes 728:739 the epitopes of all mAbs 728:751 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 4 27 theme DS 645:646 arg1 domain 649:654 the collagen-binding discoidin (DS) domain 613:654 the collagen-binding discoidin (DS) domain 613:654 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 5 28 theme collagen-binding 809:824 arg1 site 826:829 the collagen-binding site 805:829 the collagen-binding site 805:829 A conserved surface patch in the DS domain outside the collagen-binding site is shown to be required for signaling. 22483115 2 29 theme DDR 300:302 arg1 signaling 304:312 Aberrant DDR signaling 291:312 Aberrant DDR signaling 291:312 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 3 30 theme collagen 480:487 arg1 binding 489:495 collagen binding 480:495 collagen binding 480:495 We have generated monoclonal antibodies (mAbs) that inhibit DDR1 signaling without interfering with collagen binding. 22483115 0 31 theme receptor 34:41 arg1 region 59:64 the discoidin domain receptor 1 extracellular region 13:64 the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment 13:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 4 32 theme Fab 576:578 arg1 fragment 580:587 the Fab fragment 572:587 the Fab fragment of mAb 3E3 572:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 0 33 theme domain 27:32 arg1 receptor 34:41 the discoidin domain receptor 1 13:43 the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment 13:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 4 34 theme extracellular 542:554 arg1 region 556:561 the monomeric DDR1 extracellular region 523:561 the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 523:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 4 35 theme discoidin 634:642 arg1 domain 649:654 the collagen-binding discoidin (DS) domain 613:654 the collagen-binding discoidin (DS) domain 613:654 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 6 36 theme DDR1 907:910 arg1 dimer 912:916 the DDR1 dimer 903:916 the DDR1 dimer 903:916 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 0 37 theme extracellular 45:57 arg1 region 59:64 the discoidin domain receptor 1 extracellular region 13:64 the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment 13:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 4 38 theme DDR1 537:540 arg1 region 556:561 the monomeric DDR1 extracellular region 523:561 the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 523:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 5 39 theme DS 787:788 arg1 domain 790:795 the DS domain 783:795 the DS domain 783:795 A conserved surface patch in the DS domain outside the collagen-binding site is shown to be required for signaling. 22483115 6 40 theme dimer 912:916 arg1 conformation 887:898 the active conformation 876:898 the active conformation of the DDR1 dimer 876:916 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 4 41 theme region 556:561 arg1 structure 510:518 The crystal structure 498:518 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 498:598 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 6 42 theme collagen-induced 927:942 arg1 contacts 944:951 collagen-induced contacts 927:951 collagen-induced contacts between the DS domains 927:974 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 2 43 theme human 337:341 arg1 pathologies 343:353 several human pathologies 329:353 several human pathologies 329:353 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 2 43 theme human 337:341 arg1 cancers 371:377 many cancers 366:377 many cancers 366:377 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 1 44 theme dimeric 210:216 arg1 receptors 165:173 The discoidin domain receptors 144:173 The discoidin domain receptors 144:173 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 44 theme dimeric 210:216 arg1 kinases 236:242 constitutively dimeric receptor tyrosine kinases 195:242 constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen 195:288 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 2 45 theme several 329:335 arg1 pathologies 343:353 several human pathologies 329:353 several human pathologies 329:353 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 2 45 theme several 329:335 arg1 cancers 371:377 many cancers 366:377 many cancers 366:377 Aberrant DDR signaling contributes to several human pathologies, including many cancers. 22483115 1 46 theme receptor 218:225 arg1 receptors 165:173 The discoidin domain receptors 144:173 The discoidin domain receptors 144:173 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 46 theme receptor 218:225 arg1 kinases 236:242 constitutively dimeric receptor tyrosine kinases 195:242 constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen 195:288 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 6 47 theme helices 1047:1053 arg1 association 1018:1028 the previously identified association 992:1028 the previously identified association of transmembrane helices 992:1053 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 4 48 theme following 687:695 arg1 domain 705:710 the following DS-like domain 683:710 the following DS-like domain 683:710 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 22483115 1 49 theme tyrosine 227:234 arg1 receptors 165:173 The discoidin domain receptors 144:173 The discoidin domain receptors 144:173 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 1 49 theme tyrosine 227:234 arg1 kinases 236:242 constitutively dimeric receptor tyrosine kinases 195:242 constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen 195:288 The discoidin domain receptors, DDR1 and DDR2, are constitutively dimeric receptor tyrosine kinases that are activated by triple-helical collagen. 22483115 0 50 theme region 59:64 arg1 Structure 0:8 Structure 0:8 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment 0:100 Structure of the discoidin domain receptor 1 extracellular region bound to an inhibitory Fab fragment reveals features important for signaling. 22483115 5 51 from patch 774:778 arg1 domain 790:795 the DS domain 783:795 the DS domain 783:795 A conserved surface patch in the DS domain outside the collagen-binding site is shown to be required for signaling. 22483115 6 52 theme identified 1007:1016 arg1 association 1018:1028 the previously identified association 992:1028 the previously identified association of transmembrane helices 992:1053 Thus, the active conformation of the DDR1 dimer involves collagen-induced contacts between the DS domains, in addition to the previously identified association of transmembrane helices. 22483115 4 53 theme collagen-binding 617:632 arg1 domain 649:654 the collagen-binding discoidin (DS) domain 613:654 the collagen-binding discoidin (DS) domain 613:654 The crystal structure of the monomeric DDR1 extracellular region bound to the Fab fragment of mAb 3E3 reveals that the collagen-binding discoidin (DS) domain is tightly associated with the following DS-like domain, which contains the epitopes of all mAbs. 16036915 6 0 theme immunoaffinity 1161:1174 arg1 column 1176:1181 an immunoaffinity column 1158:1181 an immunoaffinity column 1158:1181 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 2 1 from mutations 296:304 arg1 ELOVL4 309:314 ELOVL4 309:314 ELOVL4 309:314 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 4 2 contain containing 731:740 arg1 ELOVL4 724:729 Wild type ELOVL4 714:729 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence 714:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 4 2 contain containing 731:740 arg2 sequence 777:784 an endoplasmic reticulum retention sequence 742:784 an endoplasmic reticulum retention sequence 742:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 7 3 theme mutant 1320:1325 arg1 ELOVL4 1327:1332 mutant ELOVL4 1320:1332 mutant ELOVL4 1320:1332 These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. 16036915 3 4 dep localization 567:578 arg1 the 551:553 the 551:553 the 551:553 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 2 5 theme Protein 277:283 arg1 mutations 296:304 Protein truncation mutations 277:304 Protein truncation mutations in ELOVL4 277:314 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 5 6 theme large 926:930 arg1 inclusions 932:941 large inclusions 926:941 large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells 926:1029 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 6 7 theme wild 1041:1044 arg1 type 1046:1049 the wild type 1037:1049 the wild type 1037:1049 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 1 8 dep acids 165:169 arg1 4 171:171 4 171:171 4 171:171 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 3 9 theme subcellular 555:565 arg1 localization 567:578 subcellular localization 555:578 subcellular localization 555:578 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 5 10 theme C-terminal 874:883 arg1 mutants 903:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 3 11 theme type 604:607 arg1 interaction 584:594 interaction 584:594 interaction 584:594 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 3 11 theme type 604:607 arg1 localization 567:578 subcellular localization 555:578 subcellular localization 555:578 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 5 12 theme ELOVL4 896:901 arg1 mutants 903:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 8 13 theme negative 1557:1564 arg1 effect 1566:1571 a dominant negative effect 1546:1571 a dominant negative effect 1546:1571 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 8 14 theme type 1581:1584 arg1 ELOVL4 1586:1591 wild type ELOVL4 1576:1591 wild type ELOVL4 1576:1591 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 3 15 theme mutant 613:618 arg1 ELOVL4 620:625 mutant ELOVL4 613:625 mutant ELOVL4 613:625 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 1 16 theme chain 153:157 arg1 acids 165:169 very long chain fatty acids 4 143:171 very long chain fatty acids 4 143:171 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 4 17 theme Wild 714:717 arg1 ELOVL4 724:729 Wild type ELOVL4 714:729 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence 714:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 7 18 theme wild 1306:1309 arg1 type 1311:1314 wild type 1306:1314 wild type 1306:1314 These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. 16036915 1 19 theme acids 165:169 arg1 elongation 129:138 elongation 129:138 elongation of very long chain fatty acids 4 129:171 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 2 20 theme Stargardt-like 373:386 arg1 degeneration 396:407 autosomal dominant Stargardt-like macular degeneration 354:407 autosomal dominant Stargardt-like macular degeneration 354:407 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 7 21 theme mutant 1433:1438 arg1 ELOVL4 1440:1445 individually expressed mutant ELOVL4 1410:1445 individually expressed mutant ELOVL4 1410:1445 These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. 16036915 4 22 theme reticulum 757:765 arg1 sequence 777:784 an endoplasmic reticulum retention sequence 742:784 an endoplasmic reticulum retention sequence 742:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 2 23 theme autosomal 354:362 arg1 degeneration 396:407 autosomal dominant Stargardt-like macular degeneration 354:407 autosomal dominant Stargardt-like macular degeneration 354:407 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 0 24 theme Dominant 0:7 arg1 mechanism 18:26 Dominant negative mechanism 0:26 Dominant negative mechanism 0:26 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 5 25 theme COS-7 1019:1023 arg1 cells 1025:1029 COS-7 cells 1019:1029 COS-7 cells 1019:1029 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 3 26 theme 293T 644:647 arg1 cells 658:662 COS-7 and HEK 293T cultured cells 630:662 COS-7 and HEK 293T cultured cells 630:662 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 1 27 theme chain 258:262 arg1 acids 270:274 very long chain fatty acids 248:274 very long chain fatty acids 248:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 6 28 theme type 1112:1115 arg1 ELOVL4 1117:1122 wild type ELOVL4 1107:1122 wild type ELOVL4 1107:1122 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 8 29 theme disease-linked 1485:1498 arg1 mutants 1522:1528 disease-linked C-terminal truncation mutants 1485:1528 disease-linked C-terminal truncation mutants of ELOVL4 1485:1538 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 0 30 theme autosomal 38:46 arg1 dystrophy 80:88 autosomal dominant Stargardt-like macular dystrophy 38:88 autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4 38:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 1 31 theme acids 270:274 arg1 biosynthesis 232:243 the biosynthesis 228:243 the biosynthesis of very long chain fatty acids 228:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 8 32 link disease-linked 1485:1498 arg1 mutants 1522:1528 disease-linked C-terminal truncation mutants 1485:1528 disease-linked C-terminal truncation mutants of ELOVL4 1485:1538 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 7 33 theme subcellular 1375:1385 arg1 localization 1387:1398 an abnormal subcellular localization 1363:1398 an abnormal subcellular localization found for individually expressed mutant ELOVL4 1363:1445 These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. 16036915 8 34 theme truncation 1511:1520 arg1 mutants 1522:1528 disease-linked C-terminal truncation mutants 1485:1528 disease-linked C-terminal truncation mutants of ELOVL4 1485:1538 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 3 35 theme autosomal 484:492 arg1 pattern 515:521 the autosomal dominant inheritance pattern 480:521 the autosomal dominant inheritance pattern of this disease 480:537 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 0 36 theme Stargardt-like 57:70 arg1 dystrophy 80:88 autosomal dominant Stargardt-like macular dystrophy 38:88 autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4 38:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 3 37 dep dominant 433:440 arg1 negative 442:449 negative 442:449 negative 442:449 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 9 38 theme autosomal 1685:1693 arg1 inheritance 1704:1714 the autosomal dominant inheritance 1681:1714 the autosomal dominant inheritance of Stargardt-like macular dystrophy 1681:1750 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 9 39 theme Stargardt-like 1719:1732 arg1 dystrophy 1742:1750 Stargardt-like macular dystrophy 1719:1750 Stargardt-like macular dystrophy 1719:1750 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 1 40 theme family 197:202 arg1 member 179:184 a member 177:184 a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids 177:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 1 40 theme family 197:202 arg1 ELOVL4 121:126 ELOVL4 121:126 ELOVL4 (elongation of very long chain fatty acids 4) 121:172 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 2 41 with patients 340:347 arg1 degeneration 396:407 autosomal dominant Stargardt-like macular degeneration 354:407 autosomal dominant Stargardt-like macular degeneration 354:407 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 6 42 theme wild 1107:1110 arg1 type 1112:1115 wild type 1107:1115 wild type ELOVL4 1107:1122 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 1 43 theme proteins 207:214 arg1 family 197:202 the ELO family 189:202 the ELO family of proteins involved in the biosynthesis of very long chain fatty acids 189:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 3 44 theme inheritance 503:513 arg1 pattern 515:521 the autosomal dominant inheritance pattern 480:521 the autosomal dominant inheritance pattern of this disease 480:537 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 5 45 theme aggresome-like 954:967 arg1 characteristics 969:983 aggresome-like characteristics 954:983 aggresome-like characteristics 954:983 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 3 46 theme disease 531:537 arg1 pattern 515:521 the autosomal dominant inheritance pattern 480:521 the autosomal dominant inheritance pattern of this disease 480:537 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 6 47 theme adjacent 1253:1260 arg1 inclusions 1242:1251 aggresome-like inclusions 1227:1251 aggresome-like inclusions adjacent to the nucleus 1227:1275 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 2 48 theme truncation 285:294 arg1 mutations 296:304 Protein truncation mutations 277:304 Protein truncation mutations in ELOVL4 277:314 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 6 49 theme aggresome-like 1227:1240 arg1 inclusions 1242:1251 aggresome-like inclusions 1227:1251 aggresome-like inclusions adjacent to the nucleus 1227:1275 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 9 50 theme dystrophy 1742:1750 arg1 inheritance 1704:1714 the autosomal dominant inheritance 1681:1714 the autosomal dominant inheritance of Stargardt-like macular dystrophy 1681:1750 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 5 51 theme disease-associated 855:872 arg1 mutants 903:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 0 52 from mutations 100:108 arg1 ELOVL4 113:118 ELOVL4 113:118 ELOVL4 113:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 9 53 theme dominant 1638:1645 arg1 mechanism 1656:1664 This dominant negative mechanism 1633:1664 This dominant negative mechanism 1633:1664 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 3 54 theme wild 599:602 arg1 type 604:607 wild type 599:607 wild type 599:607 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 5 55 theme truncation 885:894 arg1 mutants 903:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 disease-associated C-terminal truncation ELOVL4 mutants 855:909 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 8 56 theme wild 1576:1579 arg1 ELOVL4 1586:1591 wild type ELOVL4 1576:1591 wild type ELOVL4 1576:1591 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 4 57 theme type 719:722 arg1 ELOVL4 724:729 Wild type ELOVL4 714:729 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence 714:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 3 58 theme ELOVL4 620:625 arg1 interaction 584:594 interaction 584:594 interaction 584:594 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 3 58 theme ELOVL4 620:625 arg1 localization 567:578 subcellular localization 555:578 subcellular localization 555:578 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 5 59 theme juxtanuclear 990:1001 arg1 position 1003:1010 a juxtanuclear position 988:1010 a juxtanuclear position within COS-7 cells 988:1029 In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. 16036915 3 60 from localization 567:578 arg1 cells 658:662 COS-7 and HEK 293T cultured cells 630:662 COS-7 and HEK 293T cultured cells 630:662 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 1 61 theme long 148:151 arg1 acids 165:169 very long chain fatty acids 4 143:171 very long chain fatty acids 4 143:171 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 6 62 theme incultured 1089:1098 arg1 cells 1100:1104 incultured cells 1089:1104 incultured cells 1089:1104 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 1 63 dep ELOVL4 121:126 arg1 elongation 129:138 elongation 129:138 elongation of very long chain fatty acids 4 129:171 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 1 64 theme fatty 159:163 arg1 acids 165:169 very long chain fatty acids 4 143:171 very long chain fatty acids 4 143:171 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 8 65 theme subcellular 1607:1617 arg1 localization 1619:1630 its subcellular localization 1603:1630 its subcellular localization 1603:1630 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 8 66 theme dominant 1548:1555 arg1 effect 1566:1571 a dominant negative effect 1546:1571 a dominant negative effect 1546:1571 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 9 67 theme negative 1647:1654 arg1 mechanism 1656:1664 This dominant negative mechanism 1633:1664 This dominant negative mechanism 1633:1664 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 6 68 theme mutant 1055:1060 arg1 proteins 1062:1069 mutant proteins 1055:1069 mutant proteins 1055:1069 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 7 69 theme expressed 1423:1431 arg1 ELOVL4 1440:1445 individually expressed mutant ELOVL4 1410:1445 individually expressed mutant ELOVL4 1410:1445 These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. 16036915 2 70 theme dominant 364:371 arg1 degeneration 396:407 autosomal dominant Stargardt-like macular degeneration 354:407 autosomal dominant Stargardt-like macular degeneration 354:407 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 0 71 theme negative 9:16 arg1 mechanism 18:26 Dominant negative mechanism 0:26 Dominant negative mechanism 0:26 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 6 72 theme mutant 1141:1146 arg1 ELOVL4 1148:1153 mutant ELOVL4 1141:1153 mutant ELOVL4 1141:1153 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 4 73 theme endoplasmic 745:755 arg1 reticulum 757:765 an endoplasmic reticulum 742:765 an endoplasmic reticulum retention sequence 742:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 3 74 from interaction 584:594 arg1 cells 658:662 COS-7 and HEK 293T cultured cells 630:662 COS-7 and HEK 293T cultured cells 630:662 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 1 75 theme long 253:256 arg1 acids 270:274 very long chain fatty acids 248:274 very long chain fatty acids 248:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 9 76 theme dominant 1695:1702 arg1 inheritance 1704:1714 the autosomal dominant inheritance 1681:1714 the autosomal dominant inheritance of Stargardt-like macular dystrophy 1681:1750 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 3 77 theme HEK 640:642 arg1 293T 644:647 HEK 293T 640:647 HEK 293T 640:647 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 4 78 theme retention 767:775 arg1 sequence 777:784 an endoplasmic reticulum retention sequence 742:784 an endoplasmic reticulum retention sequence 742:784 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 0 79 theme dominant 48:55 arg1 dystrophy 80:88 autosomal dominant Stargardt-like macular dystrophy 38:88 autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4 38:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 1 80 theme fatty 264:268 arg1 acids 270:274 very long chain fatty acids 248:274 very long chain fatty acids 248:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 7 81 theme abnormal 1366:1373 arg1 localization 1387:1398 an abnormal subcellular localization 1363:1398 an abnormal subcellular localization found for individually expressed mutant ELOVL4 1363:1445 These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. 16036915 3 82 theme COS-7 630:634 arg1 cells 658:662 COS-7 and HEK 293T cultured cells 630:662 COS-7 and HEK 293T cultured cells 630:662 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 2 83 theme macular 388:394 arg1 degeneration 396:407 autosomal dominant Stargardt-like macular degeneration 354:407 autosomal dominant Stargardt-like macular degeneration 354:407 Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. 16036915 0 84 theme macular 72:78 arg1 dystrophy 80:88 autosomal dominant Stargardt-like macular dystrophy 38:88 autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4 38:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 9 85 theme macular 1734:1740 arg1 dystrophy 1742:1750 Stargardt-like macular dystrophy 1719:1750 Stargardt-like macular dystrophy 1719:1750 This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. 16036915 0 86 attach linked 90:95 arg2 dystrophy 80:88 autosomal dominant Stargardt-like macular dystrophy 38:88 autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4 38:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 0 86 attach linked 90:95 arg1 mutations 100:108 mutations 100:108 mutations in ELOVL4 100:118 Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. 16036915 6 87 theme mutant 1209:1214 arg1 protein 1216:1222 the mutant protein 1205:1222 the mutant protein 1205:1222 When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co-purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. 16036915 8 88 theme C-terminal 1500:1509 arg1 mutants 1522:1528 disease-linked C-terminal truncation mutants 1485:1528 disease-linked C-terminal truncation mutants of ELOVL4 1485:1538 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 16036915 3 89 theme dominant 494:501 arg1 pattern 515:521 the autosomal dominant inheritance pattern 480:521 the autosomal dominant inheritance pattern of this disease 480:537 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 3 90 theme cultured 649:656 arg1 cells 658:662 COS-7 and HEK 293T cultured cells 630:662 COS-7 and HEK 293T cultured cells 630:662 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 4 91 theme endoplasmic 807:817 arg1 reticulum 819:827 the endoplasmic reticulum 803:827 the endoplasmic reticulum 803:827 Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. 16036915 1 92 theme ELO 193:195 arg1 family 197:202 the ELO family 189:202 the ELO family of proteins involved in the biosynthesis of very long chain fatty acids 189:274 ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. 16036915 3 93 theme dominant 433:440 arg1 mechanism 451:459 a dominant negative mechanism 431:459 a dominant negative mechanism 431:459 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 3 93 theme dominant 433:440 arg1 responsible 464:474 responsible 464:474 responsible 464:474 To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. 16036915 8 94 theme ELOVL4 1533:1538 arg1 mutants 1522:1528 disease-linked C-terminal truncation mutants 1485:1528 disease-linked C-terminal truncation mutants of ELOVL4 1485:1538 From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. 11290788 3 0 theme growth 643:648 arg1 CUB-EGF 658:664 CUB-EGF 658:664 CUB-EGF 658:664 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 0 theme growth 643:648 arg1 factor 650:655 bone morphogenetic protein-1-epidermal growth factor 604:655 bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) 604:665 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 8 1 theme nM 1525:1526 arg1 values 1436:1441 K:(D) values 1430:1441 K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF) 1430:1543 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 4 2 theme Ca 961:962 arg1 presence 949:956 the presence 945:956 the presence of Ca(2+) 945:966 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 0 3 theme serine 71:76 arg1 proteases-1 78:88 serine proteases-1 71:88 serine proteases-1 71:88 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 3 4 theme bone 604:607 arg1 CUB-EGF 658:664 CUB-EGF 658:664 CUB-EGF 658:664 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 4 theme bone 604:607 arg1 factor 650:655 bone morphogenetic protein-1-epidermal growth factor 604:655 bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) 604:665 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 1 5 theme mannan-binding 138:151 arg1 MBL 161:163 MBL 161:163 MBL 161:163 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 1 5 theme mannan-binding 138:151 arg1 lectin 153:158 mannan-binding lectin 138:158 The mannan-binding lectin (MBL) activation pathway of complement 134:197 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 3 6 theme human 772:776 arg1 serum 778:782 human serum 772:782 human serum 772:782 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 7 theme morphogenetic 609:621 arg1 CUB-EGF 658:664 CUB-EGF 658:664 CUB-EGF 658:664 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 7 theme morphogenetic 609:621 arg1 factor 650:655 bone morphogenetic protein-1-epidermal growth factor 604:655 bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) 604:665 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 4 8 theme Sedimentation 785:797 arg1 analysis 808:815 Sedimentation velocity analysis 785:815 Sedimentation velocity analysis 785:815 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 8 9 theme MASP-1 1529:1534 arg1 nM 1525:1526 25.7 nM 1520:1526 25.7 nM (MASP-1 CUB-EGF) 1520:1543 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 8 9 theme MASP-1 1529:1534 arg1 CUB-EGF 1536:1542 MASP-1 CUB-EGF 1529:1542 MASP-1 CUB-EGF 1529:1542 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 6 10 from contrast 1074:1081 arg1 sensitive 1121:1129 sensitive 1121:1129 sensitive 1121:1129 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 6 10 from contrast 1074:1081 arg1 homodimer 1103:1111 the MASP-1 CUB-EGF homodimer 1084:1111 the MASP-1 CUB-EGF homodimer 1084:1111 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 1 11 theme innate 230:235 arg1 defense 244:250 the innate immune defense 226:250 the innate immune defense against pathogenic microorganisms 226:284 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 4 12 theme CUB-EGF 854:860 arg1 segments 862:869 the MASP-1 and MASP-2 CUB-EGF segments 832:869 segments 862:869 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 3 13 theme proteins 495:502 arg1 properties 475:484 the interaction properties 459:484 the interaction properties of these proteins 459:502 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 2 14 dep proteases 329:337 arg1 MASP-2 348:353 MASP-2 348:353 MASP-2 348:353 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 2 14 dep proteases 329:337 arg1 MASP-1 340:345 MASP-1 340:345 MASP-1 340:345 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 1 15 theme immune 237:242 arg1 defense 244:250 the innate immune defense 226:250 the innate immune defense against pathogenic microorganisms 226:284 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 0 16 theme MBL-associated 98:111 arg1 protein 113:119 MBL-associated protein 19 98:122 MBL-associated protein 19 98:122 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 7 17 theme gel 1244:1246 arg1 filtration 1248:1257 gel filtration 1244:1257 gel filtration 1244:1257 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 8 18 dep nM 1485:1486 arg1 MAp19 1489:1493 MAp19 1489:1493 MAp19 1489:1493 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 8 18 dep nM 1485:1486 arg1 CUB-EGF 1506:1512 MASP-2 CUB-EGF 1499:1512 MASP-2 CUB-EGF 1499:1512 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 7 19 theme resonance 1279:1287 arg1 spectroscopy 1289:1300 surface plasmon resonance spectroscopy 1263:1300 surface plasmon resonance spectroscopy 1263:1300 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 1 20 theme lectin 153:158 arg1 pathway 177:183 The mannan-binding lectin (MBL) activation pathway 134:183 The mannan-binding lectin (MBL) activation pathway of complement 134:197 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 6 21 from sensitive 1121:1129 arg1 contrast 1074:1081 contrast 1074:1081 contrast 1074:1081 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 3 22 theme factor 650:655 arg1 segments 667:674 C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments 585:674 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 2 23 located found 399:403 arg2 MAp19 387:391 MAp19 387:391 MAp19 387:391 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 2 23 located found 399:403 arg2 proteases 329:337 two MBL-associated serine proteases 303:337 two MBL-associated serine proteases (MASP-1, MASP-2) 303:354 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 2 23 located found 399:403 arg2 protein 375:381 MBL-associated protein 19 360:384 MBL-associated protein 19 (MAp19) 360:392 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 2 23 located found 399:403 arg1 serum 296:300 human serum 290:300 human serum 290:300 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 3 24 dep view 439:442 arg1 investigate 447:457 investigate 447:457 to investigate the interaction properties of these proteins 444:502 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 10 25 theme CUB-EGF 1772:1778 arg1 pair 1780:1783 the CUB-EGF pair 1768:1783 the CUB-EGF pair of each protein 1768:1799 These data indicate that MASP-1, MASP-2, and MAp19 each associate as homodimers, and individually form Ca(2+)-dependent complexes with MBL through the CUB-EGF pair of each protein. 11290788 3 26 theme MASP-2 690:695 arg1 segments 667:674 C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments 585:674 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 11 27 theme distinct 1821:1828 arg1 complexes 1839:1847 distinct MBL/MASP complexes 1821:1847 distinct MBL/MASP complexes 1821:1847 This suggests that distinct MBL/MASP complexes may be involved in the activation or regulation of the MBL pathway. 11290788 5 28 theme physical 1036:1043 arg1 properties 1045:1054 their physical properties 1030:1054 their physical properties 1030:1054 Although the latter two dimers were not dissociated by EDTA, their physical properties were affected. 11290788 7 29 theme surface 1263:1269 arg1 resonance 1279:1287 surface plasmon resonance 1263:1287 surface plasmon resonance spectroscopy 1263:1300 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 11 30 theme pathway 1908:1914 arg1 activation 1872:1881 activation 1872:1881 activation 1872:1881 This suggests that distinct MBL/MASP complexes may be involved in the activation or regulation of the MBL pathway. 11290788 11 30 theme pathway 1908:1914 arg1 regulation 1886:1895 regulation 1886:1895 regulation 1886:1895 This suggests that distinct MBL/MASP complexes may be involved in the activation or regulation of the MBL pathway. 11290788 0 31 theme Interaction 0:10 arg1 properties 12:21 Interaction properties 0:21 Interaction properties of human mannan-binding lectin (MBL) 0:58 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 3 32 theme MASP-1 679:684 arg1 segments 667:674 C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments 585:674 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 8 33 dep shown 1391:1395 arg1 each 1386:1389 each 1386:1389 each 1386:1389 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 2 34 theme human 290:294 arg1 serum 296:300 human serum 290:300 human serum 290:300 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 0 35 theme human 26:30 arg1 lectin 47:52 human mannan-binding lectin 26:52 human mannan-binding lectin (MBL) 26:58 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 0 35 theme human 26:30 arg1 MBL 55:57 MBL 55:57 MBL 55:57 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 8 36 theme CUB-EGF 1353:1359 arg1 segments 1361:1368 their CUB-EGF segments 1347:1368 their CUB-EGF segments 1347:1368 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 1 37 theme activation 166:175 arg1 pathway 177:183 The mannan-binding lectin (MBL) activation pathway 134:183 The mannan-binding lectin (MBL) activation pathway of complement 134:197 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 7 38 theme plasmon 1271:1277 arg1 resonance 1279:1287 surface plasmon resonance 1263:1287 surface plasmon resonance spectroscopy 1263:1300 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 1 39 theme pathogenic 260:269 arg1 microorganisms 271:284 pathogenic microorganisms 260:284 pathogenic microorganisms 260:284 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 8 40 theme latter 1313:1318 arg1 technique 1320:1328 the latter technique 1309:1328 the latter technique 1309:1328 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 3 41 theme human 505:509 arg1 MASP-1 511:516 human MASP-1 505:516 human MASP-1 505:516 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 10 42 theme protein 1793:1799 arg1 pair 1780:1783 the CUB-EGF pair 1768:1783 the CUB-EGF pair of each protein 1768:1799 These data indicate that MASP-1, MASP-2, and MAp19 each associate as homodimers, and individually form Ca(2+)-dependent complexes with MBL through the CUB-EGF pair of each protein. 11290788 0 43 theme lectin 47:52 arg1 properties 12:21 Interaction properties 0:21 Interaction properties of human mannan-binding lectin (MBL) 0:58 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 4 44 theme MASP-2 847:852 arg1 segments 862:869 the MASP-1 and MASP-2 CUB-EGF segments 832:869 segments 862:869 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 3 45 theme C1r/C1s 585:591 arg1 segments 667:674 C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments 585:674 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 8 46 theme nM 1467:1468 arg1 values 1436:1441 K:(D) values 1430:1441 K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF) 1430:1543 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 11 47 theme MBL/MASP 1830:1837 arg1 complexes 1839:1847 distinct MBL/MASP complexes 1821:1847 distinct MBL/MASP complexes 1821:1847 This suggests that distinct MBL/MASP complexes may be involved in the activation or regulation of the MBL pathway. 11290788 0 48 theme mannan-binding 32:45 arg1 lectin 47:52 human mannan-binding lectin 26:52 human mannan-binding lectin (MBL) 26:58 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 0 48 theme mannan-binding 32:45 arg1 MBL 55:57 MBL 55:57 MBL 55:57 Interaction properties of human mannan-binding lectin (MBL)-associated serine proteases-1 and -2, MBL-associated protein 19, and MBL. 11290788 3 49 theme N-terminal 549:558 arg1 subcomponents 571:583 the N-terminal complement subcomponents 545:583 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 50 dep subcomponents 571:583 arg1 segments 667:674 C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments 585:674 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 1 51 theme complement 188:197 arg1 pathway 177:183 The mannan-binding lectin (MBL) activation pathway 134:183 The mannan-binding lectin (MBL) activation pathway of complement 134:197 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 8 52 theme immobilized 1408:1418 arg1 MBL 1420:1422 immobilized MBL 1408:1422 immobilized MBL 1408:1422 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 3 53 theme complement 560:569 arg1 subcomponents 571:583 the N-terminal complement subcomponents 545:583 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 6 54 theme MASP-1 1088:1093 arg1 sensitive 1121:1129 sensitive 1121:1129 sensitive 1121:1129 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 6 54 theme MASP-1 1088:1093 arg1 homodimer 1103:1111 the MASP-1 CUB-EGF homodimer 1084:1111 the MASP-1 CUB-EGF homodimer 1084:1111 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 5 55 theme latter 982:987 arg1 dimers 993:998 the latter two dimers 978:998 the latter two dimers 978:998 Although the latter two dimers were not dissociated by EDTA, their physical properties were affected. 11290788 2 56 theme serine 322:327 arg1 proteases 329:337 two MBL-associated serine proteases 303:337 two MBL-associated serine proteases (MASP-1, MASP-2) 303:354 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 2 57 theme MBL-associated 360:373 arg1 protein 375:381 MBL-associated protein 19 360:384 MBL-associated protein 19 (MAp19) 360:392 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 2 57 theme MBL-associated 360:373 arg1 MAp19 387:391 MAp19 387:391 MAp19 387:391 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 4 58 theme velocity 799:806 arg1 analysis 808:815 Sedimentation velocity analysis 785:815 Sedimentation velocity analysis 785:815 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 4 59 theme homologous 879:888 arg1 MAp19 898:902 the homologous protein MAp19 875:902 the homologous protein MAp19 875:902 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 2 60 theme MBL-associated 307:320 arg1 proteases 329:337 two MBL-associated serine proteases 303:337 two MBL-associated serine proteases (MASP-1, MASP-2) 303:354 In human serum, two MBL-associated serine proteases (MASP-1, MASP-2) and MBL-associated protein 19 (MAp19) were found to be associated with MBL. 11290788 10 61 theme -dependent 1730:1739 arg1 homodimers 1690:1699 homodimers 1690:1699 homodimers 1690:1699 These data indicate that MASP-1, MASP-2, and MAp19 each associate as homodimers, and individually form Ca(2+)-dependent complexes with MBL through the CUB-EGF pair of each protein. 11290788 10 61 theme -dependent 1730:1739 arg1 complexes 1741:1749 Ca(2+)-dependent complexes 1724:1749 Ca(2+)-dependent complexes 1724:1749 These data indicate that MASP-1, MASP-2, and MAp19 each associate as homodimers, and individually form Ca(2+)-dependent complexes with MBL through the CUB-EGF pair of each protein. 11290788 10 62 dep associate 1677:1685 arg1 each 1672:1675 each 1672:1675 each 1672:1675 These data indicate that MASP-1, MASP-2, and MAp19 each associate as homodimers, and individually form Ca(2+)-dependent complexes with MBL through the CUB-EGF pair of each protein. 11290788 7 63 dep proteins 1150:1157 arg1 MASP-2 1186:1191 MASP-2 1186:1191 MASP-2 1186:1191 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 7 63 dep proteins 1150:1157 arg1 proteins 1150:1157 The three proteins and full-length MASP-1 and MASP-2 1140:1191 The three proteins and full-length MASP-1 and MASP-2 1140:1191 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 7 63 dep proteins 1150:1157 arg1 MASP-1 1175:1180 full-length MASP-1 1163:1180 full-length MASP-1 1163:1180 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 6 64 theme CUB-EGF 1095:1101 arg1 sensitive 1121:1129 sensitive 1121:1129 sensitive 1121:1129 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 6 64 theme CUB-EGF 1095:1101 arg1 homodimer 1103:1111 the MASP-1 CUB-EGF homodimer 1084:1111 the MASP-1 CUB-EGF homodimer 1084:1111 In contrast, the MASP-1 CUB-EGF homodimer was not sensitive to EDTA. 11290788 8 65 theme nM 1450:1451 arg1 values 1436:1441 K:(D) values 1430:1441 K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF) 1430:1543 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 11 66 dep activation 1872:1881 arg1 the 1868:1870 the 1868:1870 the 1868:1870 This suggests that distinct MBL/MASP complexes may be involved in the activation or regulation of the MBL pathway. 11290788 3 67 attach isolated 758:765 arg2 MBL 750:752 MBL 750:752 MBL 750:752 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 67 attach isolated 758:765 arg1 serum 778:782 human serum 772:782 human serum 772:782 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 8 68 theme nM 1485:1486 arg1 values 1436:1441 K:(D) values 1430:1441 K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF) 1430:1543 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 3 69 theme human 726:730 arg1 cells 739:743 insect or human kidney cells 716:743 cells 739:743 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 4 70 theme protein 890:896 arg1 MAp19 898:902 the homologous protein MAp19 875:902 the homologous protein MAp19 875:902 Sedimentation velocity analysis indicated that the MASP-1 and MASP-2 CUB-EGF segments and the homologous protein MAp19 all behaved as homodimers (2.8-3.2 S) in the presence of Ca(2+). 11290788 3 71 theme Uegf 594:597 arg1 segments 667:674 C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments 585:674 the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2 545:695 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 9 72 from EDTA 1602:1605 arg1 cases 1614:1618 all cases 1610:1618 all cases 1610:1618 The binding was Ca(2+)-dependent and fully sensitive to EDTA in all cases. 11290788 7 73 theme full-length 1163:1173 arg1 proteins 1150:1157 The three proteins and full-length MASP-1 and MASP-2 1140:1191 The three proteins and full-length MASP-1 and MASP-2 1140:1191 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 7 73 theme full-length 1163:1173 arg1 MASP-1 1175:1180 full-length MASP-1 1163:1180 full-length MASP-1 1163:1180 The three proteins and full-length MASP-1 and MASP-2 showed no interaction with each other as judged by gel filtration and surface plasmon resonance spectroscopy. 11290788 11 74 theme MBL 1904:1906 arg1 pathway 1908:1914 the MBL pathway 1900:1914 the MBL pathway 1900:1914 This suggests that distinct MBL/MASP complexes may be involved in the activation or regulation of the MBL pathway. 11290788 1 75 theme important 208:216 arg1 role 218:221 an important role 205:221 an important role 205:221 The mannan-binding lectin (MBL) activation pathway of complement plays an important role in the innate immune defense against pathogenic microorganisms. 11290788 3 76 theme kidney 732:737 arg1 cells 739:743 insect or human kidney cells 716:743 cells 739:743 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 77 theme protein-1-epidermal 623:641 arg1 CUB-EGF 658:664 CUB-EGF 658:664 CUB-EGF 658:664 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 3 77 theme protein-1-epidermal 623:641 arg1 factor 650:655 bone morphogenetic protein-1-epidermal growth factor 604:655 bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) 604:665 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 8 78 theme K 1430:1430 arg1 values 1436:1441 K:(D) values 1430:1441 K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF) 1430:1543 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 3 79 theme interaction 463:473 arg1 properties 475:484 the interaction properties 459:484 the interaction properties of these proteins 459:502 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 11290788 8 80 theme MASP-2 1499:1504 arg1 CUB-EGF 1506:1512 MASP-2 CUB-EGF 1499:1512 MASP-2 CUB-EGF 1499:1512 Using the latter technique, MASP-1, MASP-2, their CUB-EGF segments, and MAp19 were each shown to bind to immobilized MBL, with K:(D) values of 0.8 nM (MASP-2), 1.4 nM (MASP-1), 13.0 nM (MAp19 and MASP-2 CUB-EGF), and 25.7 nM (MASP-1 CUB-EGF). 11290788 3 81 theme insect 716:721 arg1 cells 739:743 insect or human kidney cells 716:743 cells 739:743 With a view to investigate the interaction properties of these proteins, human MASP-1, MASP-2, MAp19, as well as the N-terminal complement subcomponents C1r/C1s, Uegf, and bone morphogenetic protein-1-epidermal growth factor (CUB-EGF) segments of MASP-1 and MASP-2, were expressed in insect or human kidney cells, and MBL was isolated from human serum. 8216207 5 0 theme acceptor 1085:1092 arg1 acidic-Gly-Ser-Gly-acidic 1104:1128 the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1062:1128 the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1062:1128 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 5 1 dep possible 945:952 arg1 conform 1044:1050 conform 1044:1050 to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1041:1128 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 5 1 dep possible 945:952 arg1 proportion 1155:1164 a significant proportion 1141:1164 a significant proportion (approx 1141:1172 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 5 1 dep possible 945:952 arg1 It 938:939 It 938:939 It 938:939 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 5 2 theme consensus 1094:1102 arg1 acidic-Gly-Ser-Gly-acidic 1104:1128 the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1062:1128 the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1062:1128 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 9 3 theme apparent 1504:1511 arg1 consensus 1523:1531 An apparent 'acceptor consensus 1501:1531 An apparent 'acceptor consensus overlap' at Ser474 1501:1550 An apparent 'acceptor consensus overlap' at Ser474 suggests that the mechanism behind the glycosaminoglycan split of TM may involve a competition for substrate between xylosyltransferase and N-acetylgalactosaminyltransferase. 8216207 3 4 theme important 623:631 arg1 oligosaccharide 633:647 this functionally important oligosaccharide 605:647 this functionally important oligosaccharide 605:647 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 0 5 from Identification 0:13 arg1 thrombomodulin 97:110 soluble recombinant human thrombomodulin 71:110 soluble recombinant human thrombomodulin 71:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 3 6 theme soluble 518:524 arg1 TMD1 547:550 TMD1 547:550 TMD1 547:550 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 3 6 theme soluble 518:524 arg1 derivative 535:544 a soluble human TM derivative 516:544 a soluble human TM derivative (TMD1) 516:551 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 7 7 contain possess 1250:1256 arg2 substitution 1260:1271 a substitution 1258:1271 a substitution for Ser474 1258:1282 Mutants that possess a substitution for Ser474 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1. 8216207 7 7 contain possess 1250:1256 arg1 Mutants 1237:1243 Mutants 1237:1243 Mutants that possess a substitution for Ser474 1237:1282 Mutants that possess a substitution for Ser474 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1. 8216207 7 8 dep show 1284:1287 arg1 form 1329:1332 form 1329:1332 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1 1284:1370 Mutants that possess a substitution for Ser474 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1. 8216207 0 9 theme potential 113:121 arg1 regulation 123:132 potential regulation 113:132 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 0:199 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 6 10 theme glycosaminoglycan 1211:1227 arg1 moiety 1229:1234 a glycosaminoglycan moiety 1209:1234 a glycosaminoglycan moiety 1209:1234 35%) of the total TM still lacked a glycosaminoglycan moiety. 8216207 4 11 theme TMD1 744:747 arg1 domain 734:739 the Ser/Thr-rich domain 717:739 the Ser/Thr-rich domain of TMD1 717:747 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 11 theme TMD1 744:747 arg1 TMD1 744:747 TMD1 744:747 TMD1 744:747 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 0 12 theme functionality 137:149 arg1 regulation 123:132 potential regulation 113:132 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 0:199 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 4 13 theme serine 694:699 arg1 residues 701:708 four serine residues 689:708 four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly 689:793 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 8 14 theme C-terminal 1403:1412 arg1 peptide 1414:1420 a C-terminal peptide 1401:1420 a C-terminal peptide 1401:1420 Isolation and sequencing of a C-terminal peptide demonstrated that this serine is modified in the low-M(r) form of native TMD1. 8216207 3 15 theme TM 532:533 arg1 TMD1 547:550 TMD1 547:550 TMD1 547:550 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 3 15 theme TM 532:533 arg1 derivative 535:544 a soluble human TM derivative 516:544 a soluble human TM derivative (TMD1) 516:551 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 3 16 from s 599:599 arg1 protein 661:667 the core protein 652:667 the core protein 652:667 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 0 17 from site 63:66 arg1 thrombomodulin 97:110 soluble recombinant human thrombomodulin 71:110 soluble recombinant human thrombomodulin 71:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 4 18 theme primary 831:837 arg1 attachment 847:856 attachment 847:856 attachment 847:856 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 18 theme primary 831:837 arg1 site 839:842 the primary site 827:842 the primary site of attachment 827:856 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 3 19 theme derivative 535:544 arg1 mutagenesis 501:511 Site-directed mutagenesis 487:511 Site-directed mutagenesis of a soluble human TM derivative (TMD1) 487:551 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 2 20 theme r 452:452 arg1 form 455:458 a low-M(r) form 444:458 a low-M(r) form lacking this modification 444:484 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 4 21 theme attachment 847:856 arg1 attachment 847:856 attachment 847:856 attachment 847:856 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 21 theme attachment 847:856 arg1 site 839:842 the primary site 827:842 the primary site of attachment 827:856 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 22 theme Ser/Thr-rich 721:732 arg1 domain 734:739 the Ser/Thr-rich domain 717:739 the Ser/Thr-rich domain of TMD1 717:747 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 22 theme Ser/Thr-rich 721:732 arg1 TMD1 744:747 TMD1 744:747 TMD1 744:747 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 2 23 theme r 400:400 arg1 form 403:406 a high-M(r) form 391:406 a high-M(r) form containing chondroitin sulphate 391:438 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 1 24 theme anticoagulant 311:323 arg1 enzyme 325:330 an anticoagulant enzyme 308:330 an anticoagulant enzyme 308:330 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 6 25 theme TM 1193:1194 arg1 TM 1193:1194 the total TM 1183:1194 the total TM 1183:1194 35%) of the total TM still lacked a glycosaminoglycan moiety. 8216207 6 25 theme TM 1193:1194 arg1 % 1177:1177 35%) 1175:1178 35%) of the total TM 1175:1194 35%) of the total TM still lacked a glycosaminoglycan moiety. 8216207 3 26 theme core 656:659 arg1 protein 661:667 the core protein 652:667 the core protein 652:667 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 3 27 theme oligosaccharide 633:647 arg1 s 599:599 the attachment site(s) 579:600 the attachment site(s) of this functionally important oligosaccharide on the core protein 579:667 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 2 28 theme low-M 446:450 arg1 form 455:458 a low-M(r) form 444:458 a low-M(r) form lacking this modification 444:484 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 5 29 theme significant 1143:1153 arg1 proportion 1155:1164 a significant proportion 1141:1164 a significant proportion (approx 1141:1172 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 5 29 theme significant 1143:1153 arg1 It 938:939 It 938:939 It 938:939 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 0 30 theme glycosyltransferase 154:172 arg1 competition 174:184 glycosyltransferase competition 154:184 glycosyltransferase competition for serine474 154:198 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 6 31 theme total 1187:1191 arg1 TM 1193:1194 the total TM 1183:1194 the total TM 1183:1194 35%) of the total TM still lacked a glycosaminoglycan moiety. 8216207 1 32 theme endothelial 227:237 arg1 Thrombomodulin 201:214 Thrombomodulin 201:214 Thrombomodulin (TM) 201:219 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 1 32 theme endothelial 227:237 arg1 receptor 253:260 an endothelial cell thrombin receptor 224:260 an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme 224:330 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 9 33 dep consensus 1523:1531 arg1 overlap 1533:1539 overlap 1533:1539 overlap' at Ser474 1533:1550 An apparent 'acceptor consensus overlap' at Ser474 suggests that the mechanism behind the glycosaminoglycan split of TM may involve a competition for substrate between xylosyltransferase and N-acetylgalactosaminyltransferase. 8216207 2 34 theme chondroitin 419:429 arg1 sulphate 431:438 chondroitin sulphate 419:438 chondroitin sulphate 419:438 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 1 35 theme cell 239:242 arg1 Thrombomodulin 201:214 Thrombomodulin 201:214 Thrombomodulin (TM) 201:219 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 1 35 theme cell 239:242 arg1 receptor 253:260 an endothelial cell thrombin receptor 224:260 an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme 224:330 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 0 36 theme site 63:66 arg1 Identification 0:13 Identification 0:13 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 0:199 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 9 37 theme TM 1618:1619 arg1 split 1609:1613 split 1609:1613 split 1609:1613 An apparent 'acceptor consensus overlap' at Ser474 suggests that the mechanism behind the glycosaminoglycan split of TM may involve a competition for substrate between xylosyltransferase and N-acetylgalactosaminyltransferase. 8216207 0 38 from thrombomodulin 97:110 arg1 Identification 0:13 Identification 0:13 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 0:199 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 3 39 theme attachment 583:592 arg1 s 599:599 the attachment site(s) 579:600 the attachment site(s) of this functionally important oligosaccharide on the core protein 579:667 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 1 40 theme thrombin 244:251 arg1 Thrombomodulin 201:214 Thrombomodulin 201:214 Thrombomodulin (TM) 201:219 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 1 40 theme thrombin 244:251 arg1 receptor 253:260 an endothelial cell thrombin receptor 224:260 an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme 224:330 Thrombomodulin (TM) is an endothelial cell thrombin receptor that converts thrombin from a procoagulant to an anticoagulant enzyme. 8216207 0 41 theme glycosaminoglycan-attachment 34:61 arg1 site 63:66 the predominant glycosaminoglycan-attachment site 18:66 the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin 18:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 8 42 theme native 1488:1493 arg1 TMD1 1495:1498 native TMD1 1488:1498 native TMD1 1488:1498 Isolation and sequencing of a C-terminal peptide demonstrated that this serine is modified in the low-M(r) form of native TMD1. 8216207 4 43 from Ser472 930:935 arg1 attachment 916:925 attachment 916:925 attachment at Ser472 916:935 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 43 from Ser472 930:935 arg1 levels 906:911 low levels 902:911 low levels of attachment at Ser472 902:935 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 0 44 theme predominant 22:32 arg1 site 63:66 the predominant glycosaminoglycan-attachment site 18:66 the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin 18:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 0 45 theme soluble 71:77 arg1 thrombomodulin 97:110 soluble recombinant human thrombomodulin 71:110 soluble recombinant human thrombomodulin 71:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 7 46 theme increased 1292:1300 arg1 mobility 1302:1309 an increased mobility 1289:1309 an increased mobility of their low-M(r) 1289:1327 Mutants that possess a substitution for Ser474 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1. 8216207 7 47 theme low-M 1320:1324 arg1 mobility 1302:1309 an increased mobility 1289:1309 an increased mobility of their low-M(r) 1289:1327 Mutants that possess a substitution for Ser474 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1. 8216207 8 48 theme low-M 1471:1475 arg1 form 1480:1483 the low-M(r) form 1467:1483 the low-M(r) form of native TMD1 1467:1498 Isolation and sequencing of a C-terminal peptide demonstrated that this serine is modified in the low-M(r) form of native TMD1. 8216207 3 49 theme Site-directed 487:499 arg1 mutagenesis 501:511 Site-directed mutagenesis 487:511 Site-directed mutagenesis of a soluble human TM derivative (TMD1) 487:551 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 5 50 dep conform 1044:1050 arg1 as 1038:1039 as 1038:1039 as 1038:1039 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 4 51 theme low 902:904 arg1 levels 906:911 low levels 902:911 low levels of attachment at Ser472 902:935 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 2 52 contain containing 408:417 arg1 form 403:406 a high-M(r) form 391:406 a high-M(r) form containing chondroitin sulphate 391:438 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 2 52 contain containing 408:417 arg2 sulphate 431:438 chondroitin sulphate 419:438 chondroitin sulphate 419:438 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 9 53 theme acceptor 1514:1521 arg1 consensus 1523:1531 An apparent 'acceptor consensus 1501:1531 An apparent 'acceptor consensus overlap' at Ser474 1501:1550 An apparent 'acceptor consensus overlap' at Ser474 suggests that the mechanism behind the glycosaminoglycan split of TM may involve a competition for substrate between xylosyltransferase and N-acetylgalactosaminyltransferase. 8216207 8 54 theme TMD1 1495:1498 arg1 form 1480:1483 the low-M(r) form 1467:1483 the low-M(r) form of native TMD1 1467:1498 Isolation and sequencing of a C-terminal peptide demonstrated that this serine is modified in the low-M(r) form of native TMD1. 8216207 5 55 theme attachment 987:996 arg1 degree 977:982 the overall degree 965:982 the overall degree of attachment 965:996 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 3 56 theme site 594:597 arg1 s 599:599 the attachment site(s) 579:600 the attachment site(s) of this functionally important oligosaccharide on the core protein 579:667 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 7 57 theme native 1360:1365 arg1 TMD1 1367:1370 native TMD1 1360:1370 native TMD1 1360:1370 Mutants that possess a substitution for Ser474 show an increased mobility of their low-M(r) form on SDS/PAGE compared with native TMD1. 8216207 5 58 theme glutamic 1020:1027 arg1 acid 1029:1032 glutamic acid 1020:1032 glutamic acid 1020:1032 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 4 59 theme glycosaminoglycan 768:784 arg1 assembly 786:793 glycosaminoglycan assembly 768:793 glycosaminoglycan assembly 768:793 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 4 60 from levels 906:911 arg1 Ser472 930:935 Ser472 930:935 Ser472 930:935 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 8216207 0 61 theme human 91:95 arg1 thrombomodulin 97:110 soluble recombinant human thrombomodulin 71:110 soluble recombinant human thrombomodulin 71:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 8 62 theme peptide 1414:1420 arg1 sequencing 1387:1396 sequencing 1387:1396 sequencing 1387:1396 Isolation and sequencing of a C-terminal peptide demonstrated that this serine is modified in the low-M(r) form of native TMD1. 8216207 8 62 theme peptide 1414:1420 arg1 Isolation 1373:1381 Isolation 1373:1381 Isolation 1373:1381 Isolation and sequencing of a C-terminal peptide demonstrated that this serine is modified in the low-M(r) form of native TMD1. 8216207 5 63 theme overall 969:975 arg1 degree 977:982 the overall degree 965:982 the overall degree of attachment 965:996 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 2 64 theme high-M 393:398 arg1 form 403:406 a high-M(r) form 391:406 a high-M(r) form containing chondroitin sulphate 391:438 It has previously been shown that TM is expressed in both a high-M(r) form containing chondroitin sulphate and a low-M(r) form lacking this modification. 8216207 0 65 theme recombinant 79:89 arg1 thrombomodulin 97:110 soluble recombinant human thrombomodulin 71:110 soluble recombinant human thrombomodulin 71:110 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 5 66 theme xylosyltransferase 1066:1083 arg1 acidic-Gly-Ser-Gly-acidic 1104:1128 the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1062:1128 the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic 1062:1128 It was possible to improve the overall degree of attachment by mutating Ser472 to glutamic acid (so as to conform Ser474 to the xylosyltransferase acceptor consensus acidic-Gly-Ser-Gly-acidic); however, a significant proportion (approx. 8216207 0 67 dep Identification 0:13 arg1 regulation 123:132 potential regulation 113:132 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 0:199 Identification of the predominant glycosaminoglycan-attachment site in soluble recombinant human thrombomodulin: potential regulation of functionality by glycosyltransferase competition for serine474. 8216207 9 68 dep mechanism 1570:1578 arg1 behind 1580:1585 behind 1580:1585 behind the glycosaminoglycan split of TM 1580:1619 An apparent 'acceptor consensus overlap' at Ser474 suggests that the mechanism behind the glycosaminoglycan split of TM may involve a competition for substrate between xylosyltransferase and N-acetylgalactosaminyltransferase. 8216207 3 69 theme human 526:530 arg1 TMD1 547:550 TMD1 547:550 TMD1 547:550 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 3 69 theme human 526:530 arg1 derivative 535:544 a soluble human TM derivative 516:544 a soluble human TM derivative (TMD1) 516:551 Site-directed mutagenesis of a soluble human TM derivative (TMD1) was employed to determine the attachment site(s) of this functionally important oligosaccharide on the core protein. 8216207 4 70 theme attachment 916:925 arg1 levels 906:911 low levels 902:911 low levels of attachment at Ser472 902:935 Although there are four serine residues within the Ser/Thr-rich domain of TMD1 that might support glycosaminoglycan assembly, our analysis demonstrates that the primary site of attachment is at Ser474, and evidence is presented for low levels of attachment at Ser472. 18695242 4 0 dep led 546:548 arg1 characterized 629:641 characterized 629:641 characterized by aberrant cartilage morphology 629:674 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 7 1 theme nucleotide 1102:1111 arg1 product 1113:1119 the nucleotide product 1098:1119 the nucleotide product of sulfotransferases within the Golgi 1098:1157 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 0 2 from role 2:5 arg1 sulfation 78:86 sulfation 78:86 sulfation 78:86 A role for a lithium-inhibited Golgi nucleotidase in skeletal development and sulfation. 18695242 0 2 from role 2:5 arg1 development 62:72 development 62:72 development 62:72 A role for a lithium-inhibited Golgi nucleotidase in skeletal development and sulfation. 18695242 5 3 theme phenotypic 681:690 arg1 similarities 692:703 The phenotypic similarities 677:703 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway 677:815 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 5 4 from discovery 829:837 arg1 absence 875:881 the absence 871:881 the absence of functional enzyme 871:902 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 4 5 from inactivation 516:527 arg1 mice 541:544 mice 541:544 mice 541:544 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 7 6 from function 1281:1288 arg1 formation 1307:1315 the formation 1303:1315 the formation of skeletal elements derived through endochondral ossification 1303:1378 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 2 7 theme sulfated 310:317 arg1 acceptors 319:327 sulfated acceptors 310:327 sulfated acceptors 310:327 It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). 18695242 6 8 theme whole 1028:1032 arg1 embryos 1034:1040 whole embryos 1028:1040 whole embryos 1028:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 6 9 theme lung 1012:1015 arg1 tissue 1017:1022 lung tissue 1012:1022 lung tissue 1012:1022 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 7 10 with consistent 1056:1065 arg1 model 1074:1078 a model 1072:1078 a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation 1072:1212 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 7 11 theme unique 1225:1230 arg1 basis 1240:1244 a unique genetic basis 1223:1244 a unique genetic basis for chondrodysplasia 1223:1265 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 6 12 theme gPAPP 939:943 arg1 loss 931:934 loss 931:934 loss of gPAPP 931:943 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 5 13 theme functional 886:895 arg1 enzyme 897:902 functional enzyme 886:902 functional enzyme 886:902 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 5 14 theme chondroitin 856:866 arg1 discovery 829:837 the discovery 825:837 the discovery of undersulfated chondroitin in the absence of functional enzyme 825:902 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 1 15 theme numerous 165:172 arg1 molecules 174:182 numerous molecules 165:182 numerous molecules 165:182 Sulfation is an important biological process that modulates the function of numerous molecules. 18695242 7 16 theme genetic 1232:1238 arg1 basis 1240:1244 a unique genetic basis 1223:1244 a unique genetic basis for chondrodysplasia 1223:1265 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 1 17 theme molecules 174:182 arg1 function 153:160 the function 149:160 the function of numerous molecules 149:182 Sulfation is an important biological process that modulates the function of numerous molecules. 18695242 3 18 theme PAP 408:410 arg1 gPAPP 428:432 gPAPP 428:432 gPAPP 428:432 Here, we identify a Golgi-resident PAP 3'-phosphatase (gPAPP) and demonstrate that its activity is potently inhibited by lithium in vitro. 18695242 3 18 theme PAP 408:410 arg1 3'-phosphatase 412:425 a Golgi-resident PAP 3'-phosphatase 391:425 a Golgi-resident PAP 3'-phosphatase (gPAPP) 391:433 Here, we identify a Golgi-resident PAP 3'-phosphatase (gPAPP) and demonstrate that its activity is potently inhibited by lithium in vitro. 18695242 7 19 theme product 1113:1119 arg1 clearance 1085:1093 clearance 1085:1093 clearance of the nucleotide product of sulfotransferases within the Golgi 1085:1157 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 6 20 from perturbations 954:966 arg1 levels 975:980 the levels 971:980 the levels of heparan sulfate species in lung tissue and whole embryos 971:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 6 21 from levels 975:980 arg1 embryos 1034:1040 whole embryos 1028:1040 whole embryos 1028:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 6 21 from levels 975:980 arg1 tissue 1017:1022 lung tissue 1012:1022 lung tissue 1012:1022 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 7 22 theme endochondral 1354:1365 arg1 ossification 1367:1378 endochondral ossification 1354:1378 endochondral ossification 1354:1378 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 4 23 theme gPAPP 532:536 arg1 inactivation 516:527 The inactivation 512:527 The inactivation of gPAPP in mice 512:544 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 6 24 from embryos 1034:1040 arg1 levels 975:980 the levels 971:980 the levels of heparan sulfate species in lung tissue and whole embryos 971:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 2 25 theme 3'-phosphoadenosine 333:351 arg1 PAP 367:369 PAP 367:369 PAP 367:369 It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). 18695242 2 25 theme 3'-phosphoadenosine 333:351 arg1 5'-phosphate 353:364 3'-phosphoadenosine 5'-phosphate 333:364 3'-phosphoadenosine 5'-phosphate (PAP) 333:370 It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). 18695242 4 26 theme neonatal 553:560 arg1 lethality 562:570 neonatal lethality 553:570 neonatal lethality 553:570 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 5 27 theme chondrodysplastic 729:745 arg1 models 747:752 chondrodysplastic models 729:752 chondrodysplastic models harboring mutations within components of the sulfation pathway 729:815 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 4 28 theme cartilage 655:663 arg1 morphology 665:674 aberrant cartilage morphology 646:674 aberrant cartilage morphology 646:674 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 3 29 theme Golgi-resident 393:406 arg1 gPAPP 428:432 gPAPP 428:432 gPAPP 428:432 Here, we identify a Golgi-resident PAP 3'-phosphatase (gPAPP) and demonstrate that its activity is potently inhibited by lithium in vitro. 18695242 3 29 theme Golgi-resident 393:406 arg1 3'-phosphatase 412:425 a Golgi-resident PAP 3'-phosphatase 391:425 a Golgi-resident PAP 3'-phosphatase (gPAPP) 391:433 Here, we identify a Golgi-resident PAP 3'-phosphatase (gPAPP) and demonstrate that its activity is potently inhibited by lithium in vitro. 18695242 4 30 theme lung 573:576 arg1 abnormalities 578:590 lung abnormalities 573:590 lung abnormalities resembling atelectasis, and dwarfism 573:627 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 6 31 from species 1001:1007 arg1 embryos 1034:1040 whole embryos 1028:1040 whole embryos 1028:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 6 31 from species 1001:1007 arg1 tissue 1017:1022 lung tissue 1012:1022 lung tissue 1012:1022 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 7 32 dep model 1074:1078 arg1 plays 1159:1163 plays 1159:1163 plays an important role in glycosaminoglycan sulfation 1159:1212 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 4 33 theme aberrant 646:653 arg1 morphology 665:674 aberrant cartilage morphology 646:674 aberrant cartilage morphology 646:674 The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. 18695242 0 34 theme Golgi 31:35 arg1 nucleotidase 37:48 a lithium-inhibited Golgi nucleotidase 11:48 a lithium-inhibited Golgi nucleotidase 11:48 A role for a lithium-inhibited Golgi nucleotidase in skeletal development and sulfation. 18695242 6 35 theme species 1001:1007 arg1 levels 975:980 the levels 971:980 the levels of heparan sulfate species in lung tissue and whole embryos 971:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 6 36 from tissue 1017:1022 arg1 levels 975:980 the levels 971:980 the levels of heparan sulfate species in lung tissue and whole embryos 971:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 7 37 theme skeletal 1320:1327 arg1 elements 1329:1336 skeletal elements 1320:1336 skeletal elements derived through endochondral ossification 1320:1378 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 5 38 theme enzyme 897:902 arg1 absence 875:881 the absence 871:881 the absence of functional enzyme 871:902 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 0 39 theme lithium-inhibited 13:29 arg1 nucleotidase 37:48 a lithium-inhibited Golgi nucleotidase 11:48 a lithium-inhibited Golgi nucleotidase 11:48 A role for a lithium-inhibited Golgi nucleotidase in skeletal development and sulfation. 18695242 6 40 theme sulfate 993:999 arg1 species 1001:1007 heparan sulfate species 985:1007 heparan sulfate species in lung tissue and whole embryos 985:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 7 41 theme glycosaminoglycan 1186:1202 arg1 sulfation 1204:1212 glycosaminoglycan sulfation 1186:1212 glycosaminoglycan sulfation 1186:1212 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 2 42 theme Golgi 226:230 arg1 sulfotransferases 232:248 Golgi sulfotransferases 226:248 Golgi sulfotransferases 226:248 It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). 18695242 7 43 theme elements 1329:1336 arg1 formation 1307:1315 the formation 1303:1315 the formation of skeletal elements derived through endochondral ossification 1303:1378 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 7 44 theme important 1168:1176 arg1 role 1178:1181 an important role 1165:1181 an important role 1165:1181 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 6 45 theme heparan 985:991 arg1 sulfate 993:999 heparan sulfate 985:999 heparan sulfate species in lung tissue and whole embryos 985:1040 Additionally, we observed loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. 18695242 5 46 theme gPAPP 708:712 arg1 mice 721:724 gPAPP mutant mice 708:724 gPAPP mutant mice 708:724 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 5 47 theme pathway 809:815 arg1 components 781:790 components 781:790 components of the sulfation pathway 781:815 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 1 48 theme important 105:113 arg1 process 126:132 an important biological process 102:132 an important biological process that modulates the function of numerous molecules 102:182 Sulfation is an important biological process that modulates the function of numerous molecules. 18695242 1 48 theme important 105:113 arg1 Sulfation 89:97 Sulfation 89:97 Sulfation 89:97 Sulfation is an important biological process that modulates the function of numerous molecules. 18695242 5 49 theme undersulfated 842:854 arg1 chondroitin 856:866 undersulfated chondroitin 842:866 undersulfated chondroitin 842:866 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 5 50 theme mutant 714:719 arg1 mice 721:724 gPAPP mutant mice 708:724 gPAPP mutant mice 708:724 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 1 51 theme biological 115:124 arg1 process 126:132 an important biological process 102:132 an important biological process that modulates the function of numerous molecules 102:182 Sulfation is an important biological process that modulates the function of numerous molecules. 18695242 1 51 theme biological 115:124 arg1 Sulfation 89:97 Sulfation 89:97 Sulfation 89:97 Sulfation is an important biological process that modulates the function of numerous molecules. 18695242 5 52 theme mice 721:724 arg1 similarities 692:703 The phenotypic similarities 677:703 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway 677:815 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 2 53 theme 3'-phosphoadenosine 261:279 arg1 5'-phosphosulfate 281:297 3'-phosphoadenosine 5'-phosphosulfate 261:297 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors 261:327 It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). 18695242 7 54 theme sulfotransferases 1124:1140 arg1 product 1113:1119 the nucleotide product 1098:1119 the nucleotide product of sulfotransferases within the Golgi 1098:1157 Our data are consistent with a model that clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification. 18695242 5 55 theme sulfation 799:807 arg1 pathway 809:815 the sulfation pathway 795:815 the sulfation pathway 795:815 The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. 18695242 0 56 theme skeletal 53:60 arg1 development 62:72 development 62:72 development 62:72 A role for a lithium-inhibited Golgi nucleotidase in skeletal development and sulfation. 18695242 2 57 dep 5'-phosphosulfate 281:297 arg1 produce 302:308 produce 302:308 to produce sulfated acceptors 299:327 It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). 3550808 3 0 theme sequence 303:310 arg1 Comparison 285:294 Comparison 285:294 Comparison of the sequence to other serine proteinases 285:338 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 4 1 theme binding 663:669 arg1 types 654:658 alternative types 642:658 alternative types of binding with the human proteinase 642:695 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 1 2 theme complete 52:59 arg1 sequence 72:79 The complete amino acid sequence 48:79 The complete amino acid sequence of human neutrophil elastase 48:108 The complete amino acid sequence of human neutrophil elastase has been determined. 3550808 3 3 theme moderate 355:362 arg1 homology 364:371 only moderate homology 350:371 only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%) 350:446 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 4 4 with binding 663:669 arg1 proteinase 686:695 the human proteinase 676:695 the human proteinase 676:695 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 1 5 theme amino 61:65 arg1 sequence 72:79 The complete amino acid sequence 48:79 The complete amino acid sequence of human neutrophil elastase 48:108 The complete amino acid sequence of human neutrophil elastase has been determined. 3550808 2 6 theme disulfide 268:276 arg1 bonds 278:282 four disulfide bonds 263:282 four disulfide bonds 263:282 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 1 7 theme acid 67:70 arg1 sequence 72:79 The complete amino acid sequence 48:79 The complete amino acid sequence of human neutrophil elastase 48:108 The complete amino acid sequence of human neutrophil elastase has been determined. 3550808 3 8 theme neutrophil 417:426 arg1 G 438:438 neutrophil cathepsin G 417:438 neutrophil cathepsin G (37.2%) 417:446 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 8 theme neutrophil 417:426 arg1 % 445:445 37.2% 441:445 37.2% 441:445 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 9 theme pancreatic 386:395 arg1 elastase 397:404 porcine pancreatic elastase 378:404 porcine pancreatic elastase (43.0%) 378:412 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 9 theme pancreatic 386:395 arg1 % 411:411 43.0% 407:411 43.0% 407:411 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 10 theme cathepsin 428:436 arg1 G 438:438 neutrophil cathepsin G 417:438 neutrophil cathepsin G (37.2%) 417:446 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 10 theme cathepsin 428:436 arg1 % 445:445 37.2% 441:445 37.2% 441:445 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 11 theme porcine 378:384 arg1 elastase 397:404 porcine pancreatic elastase 378:404 porcine pancreatic elastase (43.0%) 378:412 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 11 theme porcine 378:384 arg1 % 411:411 43.0% 407:411 43.0% 407:411 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 12 with homology 364:371 arg1 % 445:445 37.2% 441:445 37.2% 441:445 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 12 with homology 364:371 arg1 G 438:438 neutrophil cathepsin G 417:438 neutrophil cathepsin G (37.2%) 417:446 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 12 with homology 364:371 arg1 elastase 397:404 porcine pancreatic elastase 378:404 porcine pancreatic elastase (43.0%) 378:412 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 3 12 with homology 364:371 arg1 % 411:411 43.0% 407:411 43.0% 407:411 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 0 13 theme Primary 0:6 arg1 structure 8:16 Primary structure 0:16 Primary structure of human neutrophil elastase 0:45 Primary structure of human neutrophil elastase. 3550808 1 14 theme human 84:88 arg1 elastase 101:108 human neutrophil elastase 84:108 human neutrophil elastase 84:108 The complete amino acid sequence of human neutrophil elastase has been determined. 3550808 0 15 theme human 21:25 arg1 elastase 38:45 human neutrophil elastase 21:45 human neutrophil elastase 21:45 Primary structure of human neutrophil elastase. 3550808 4 16 theme important 503:511 arg1 roles 513:517 important roles 503:517 important roles 503:517 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 1 17 theme neutrophil 90:99 arg1 elastase 101:108 human neutrophil elastase 84:108 human neutrophil elastase 84:108 The complete amino acid sequence of human neutrophil elastase has been determined. 3550808 1 18 theme elastase 101:108 arg1 sequence 72:79 The complete amino acid sequence 48:79 The complete amino acid sequence of human neutrophil elastase 48:108 The complete amino acid sequence of human neutrophil elastase has been determined. 3550808 2 19 contain contains 180:187 arg1 protein 135:141 The protein 131:141 The protein 131:141 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 2 19 contain contains 180:187 arg2 chains 229:234 two asparagine-linked carbohydrate side chains 189:234 two asparagine-linked carbohydrate side chains 189:234 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 0 20 theme elastase 38:45 arg1 structure 8:16 Primary structure 0:16 Primary structure of human neutrophil elastase 0:45 Primary structure of human neutrophil elastase. 3550808 2 21 theme acid 165:168 arg1 residues 170:177 218 amino acid residues 155:177 218 amino acid residues 155:177 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 2 22 link asparagine-linked 193:209 arg1 chains 229:234 two asparagine-linked carbohydrate side chains 189:234 two asparagine-linked carbohydrate side chains 189:234 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 0 23 theme neutrophil 27:36 arg1 elastase 38:45 human neutrophil elastase 21:45 human neutrophil elastase 21:45 Primary structure of human neutrophil elastase. 3550808 2 24 theme amino 159:163 arg1 residues 170:177 218 amino acid residues 155:177 218 amino acid residues 155:177 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 4 25 theme residues 476:483 arg1 many 464:467 many 464:467 many 464:467 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 4 25 theme residues 476:483 arg1 residues 476:483 the residues 472:483 the residues 472:483 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 2 26 theme side 224:227 arg1 chains 229:234 two asparagine-linked carbohydrate side chains 189:234 two asparagine-linked carbohydrate side chains 189:234 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 2 27 theme asparagine-linked 193:209 arg1 chains 229:234 two asparagine-linked carbohydrate side chains 189:234 two asparagine-linked carbohydrate side chains 189:234 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 2 28 theme carbohydrate 211:222 arg1 chains 229:234 two asparagine-linked carbohydrate side chains 189:234 two asparagine-linked carbohydrate side chains 189:234 The protein consists of 218 amino acid residues, contains two asparagine-linked carbohydrate side chains, and is joined together by four disulfide bonds. 3550808 4 29 theme elastase 560:567 arg1 functions 569:577 the pancreatic elastase functions 545:577 the pancreatic elastase functions 545:577 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 4 30 theme human 680:684 arg1 proteinase 686:695 the human proteinase 676:695 the human proteinase 676:695 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 4 31 theme pancreatic 549:558 arg1 functions 569:577 the pancreatic elastase functions 545:577 the pancreatic elastase functions 545:577 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 3 32 theme other 315:319 arg1 proteinases 328:338 other serine proteinases 315:338 other serine proteinases 315:338 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 4 33 theme neutrophil 612:621 arg1 enzyme 623:628 the neutrophil enzyme 608:628 the neutrophil enzyme 608:628 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 3550808 3 34 theme serine 321:326 arg1 proteinases 328:338 other serine proteinases 315:338 other serine proteinases 315:338 Comparison of the sequence to other serine proteinases indicates only moderate homology with porcine pancreatic elastase (43.0%) or neutrophil cathepsin G (37.2%). 3550808 4 35 theme alternative 642:652 arg1 types 654:658 alternative types 642:658 alternative types of binding with the human proteinase 642:695 In particular, many of the residues suggested to play important roles in the mechanism by which the pancreatic elastase functions are significantly changed in the neutrophil enzyme, indicating alternative types of binding with the human proteinase. 8089145 0 0 theme catalytic 80:88 arg1 activity 90:97 catalytic activity 80:97 catalytic activity 80:97 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 4 1 from localization 902:913 arg1 cells 991:995 baby hamster kidney cells 971:995 baby hamster kidney cells 971:995 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 4 2 theme intracellular 888:900 arg1 localization 902:913 intracellular localization 888:913 intracellular localization 888:913 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 1 3 theme leukocyte 156:164 arg1 hydrolase 143:151 Acyloxyacyl hydrolase 131:151 Acyloxyacyl hydrolase 131:151 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 1 3 theme leukocyte 156:164 arg1 enzyme 166:171 a leukocyte enzyme 154:171 a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids 154:244 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 2 4 theme amino 539:543 arg1 similarity 559:568 amino acid sequence similarity 539:568 amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases 539:658 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 3 5 theme acyloxyacyl 684:694 arg1 subunits 706:713 both acyloxyacyl hydrolase subunits 679:713 both acyloxyacyl hydrolase subunits 679:713 We show here that both acyloxyacyl hydrolase subunits are required for catalytic activity toward LPS and glycerophosphatidylcholine. 8089145 2 6 theme activator 596:604 arg1 proteins 606:613 sphingolipid activator proteins 583:613 sphingolipid activator proteins 583:613 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 6 theme activator 596:604 arg1 saposins 573:580 saposins 573:580 saposins (sphingolipid activator proteins) 573:614 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 7 theme smaller 519:525 arg1 subunit 527:533 the smaller subunit 515:533 the smaller subunit 515:533 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 3 8 theme hydrolase 696:704 arg1 subunits 706:713 both acyloxyacyl hydrolase subunits 679:713 both acyloxyacyl hydrolase subunits 679:713 We show here that both acyloxyacyl hydrolase subunits are required for catalytic activity toward LPS and glycerophosphatidylcholine. 8089145 2 9 theme sphingolipid 583:594 arg1 proteins 606:613 sphingolipid activator proteins 583:613 sphingolipid activator proteins 583:613 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 9 theme sphingolipid 583:594 arg1 saposins 573:580 saposins 573:580 saposins (sphingolipid activator proteins) 573:614 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 3 10 theme catalytic 732:740 arg1 activity 742:749 catalytic activity 732:749 catalytic activity toward LPS and glycerophosphatidylcholine 732:791 We show here that both acyloxyacyl hydrolase subunits are required for catalytic activity toward LPS and glycerophosphatidylcholine. 8089145 2 11 located found 466:470 arg2 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 11 located found 466:470 arg1 sites 486:490 the active sites 475:490 the active sites of many lipases 475:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 11 located found 466:470 arg2 sequence 431:438 an amino acid sequence 417:438 an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases 417:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 11 located found 466:470 arg1 lipases 500:506 many lipases 495:506 many lipases 495:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 0 12 theme acyloxyacyl 108:118 arg1 hydrolase 120:128 human acyloxyacyl hydrolase 102:128 human acyloxyacyl hydrolase 102:128 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 4 13 theme profound 864:871 arg1 effects 873:879 profound effects 864:879 profound effects 864:879 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 2 14 theme sphingolipid 631:642 arg1 glycohydrolases 644:658 sphingolipid glycohydrolases 631:658 sphingolipid glycohydrolases 631:658 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 15 contain has 535:537 arg1 subunit 527:533 the smaller subunit 515:533 the smaller subunit 515:533 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 15 contain has 535:537 arg2 similarity 559:568 amino acid sequence similarity 539:568 amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases 539:658 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 0 16 theme human 102:106 arg1 hydrolase 120:128 human acyloxyacyl hydrolase 102:128 human acyloxyacyl hydrolase 102:128 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 2 17 theme active 479:484 arg1 sites 486:490 the active sites 475:490 the active sites of many lipases 475:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 17 theme active 479:484 arg1 lipases 500:506 many lipases 495:506 many lipases 495:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 6 18 theme subunits 1213:1220 arg1 orientation 1190:1200 Proper orientation 1183:1200 Proper orientation of the two subunits 1183:1220 Proper orientation of the two subunits thus seems very important for the substrate specificity of this unusual enzyme. 8089145 4 19 theme kidney 984:989 arg1 cells 991:995 baby hamster kidney cells 971:995 baby hamster kidney cells 971:995 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 0 20 theme saposin-like 2:13 arg1 domain 15:20 A saposin-like domain 0:20 A saposin-like domain 0:20 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 4 21 theme baby 971:974 arg1 kidney 984:989 baby hamster kidney 971:989 baby hamster kidney cells 971:995 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 1 22 theme precursor 267:275 arg1 hydrolase 143:151 Acyloxyacyl hydrolase 131:151 Acyloxyacyl hydrolase 131:151 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 1 22 theme precursor 267:275 arg1 polypeptide 277:287 a precursor polypeptide 265:287 a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits 265:386 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 6 23 theme enzyme 1294:1299 arg1 specificity 1266:1276 the substrate specificity 1252:1276 the substrate specificity of this unusual enzyme 1252:1299 Proper orientation of the two subunits thus seems very important for the substrate specificity of this unusual enzyme. 8089145 1 24 theme bacterial 186:194 arg1 LPSs 217:220 LPSs 217:220 LPSs 217:220 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 1 24 theme bacterial 186:194 arg1 lipopolysaccharides 196:214 bacterial lipopolysaccharides 186:214 bacterial lipopolysaccharides (LPSs) 186:221 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 2 25 theme acid 545:548 arg1 similarity 559:568 amino acid sequence similarity 539:568 amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases 539:658 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 6 26 theme unusual 1286:1292 arg1 enzyme 1294:1299 this unusual enzyme 1281:1299 this unusual enzyme 1281:1299 Proper orientation of the two subunits thus seems very important for the substrate specificity of this unusual enzyme. 8089145 0 27 theme hydrolase 120:128 arg1 stability 65:73 stability 65:73 stability 65:73 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 0 27 theme hydrolase 120:128 arg1 localization 51:62 the intracellular localization 33:62 the intracellular localization 33:62 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 0 27 theme hydrolase 120:128 arg1 activity 90:97 catalytic activity 80:97 catalytic activity 80:97 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 0 28 theme intracellular 37:49 arg1 localization 51:62 the intracellular localization 33:62 the intracellular localization 33:62 A saposin-like domain influences the intracellular localization, stability, and catalytic activity of human acyloxyacyl hydrolase. 8089145 4 29 theme enzyme 961:966 arg1 localization 902:913 intracellular localization 888:913 intracellular localization 888:913 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 4 29 theme enzyme 961:966 arg1 processing 928:937 proteolytic processing 916:937 proteolytic processing 916:937 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 4 29 theme enzyme 961:966 arg1 stability 944:952 stability 944:952 stability 944:952 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 5 30 theme precursor 1038:1046 arg1 protein 1048:1054 the precursor protein 1034:1054 the precursor protein 1034:1054 Remarkably, proteolytic cleavage of the precursor protein increases the activity of the enzyme toward LPS by 10-20-fold without altering its activity toward glycerophosphatidylcholine. 8089145 2 31 theme larger 393:398 arg1 subunit 400:406 The larger subunit 389:406 The larger subunit 389:406 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 32 theme acid 426:429 arg1 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 32 theme acid 426:429 arg1 sequence 431:438 an amino acid sequence 417:438 an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases 417:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 1 33 theme internal 304:311 arg1 linkage 323:329 internal disulfide linkage 304:329 internal disulfide linkage 304:329 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 2 34 contain contains 408:415 arg2 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 34 contain contains 408:415 arg2 sequence 431:438 an amino acid sequence 417:438 an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases 417:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 34 contain contains 408:415 arg1 subunit 400:406 The larger subunit 389:406 The larger subunit 389:406 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 5 35 theme protein 1048:1054 arg1 cleavage 1022:1029 proteolytic cleavage 1010:1029 proteolytic cleavage of the precursor protein 1010:1054 Remarkably, proteolytic cleavage of the precursor protein increases the activity of the enzyme toward LPS by 10-20-fold without altering its activity toward glycerophosphatidylcholine. 8089145 2 36 theme amino 420:424 arg1 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 Gly-X-Ser-X-Gly 441:455 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 36 theme amino 420:424 arg1 sequence 431:438 an amino acid sequence 417:438 an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases 417:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 1 37 theme disulfide 313:321 arg1 linkage 323:329 internal disulfide linkage 304:329 internal disulfide linkage 304:329 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 4 38 from stability 944:952 arg1 cells 991:995 baby hamster kidney cells 971:995 baby hamster kidney cells 971:995 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 6 39 theme Proper 1183:1188 arg1 orientation 1190:1200 Proper orientation 1183:1200 Proper orientation of the two subunits 1183:1220 Proper orientation of the two subunits thus seems very important for the substrate specificity of this unusual enzyme. 8089145 4 40 theme proteolytic 916:926 arg1 processing 928:937 proteolytic processing 916:937 proteolytic processing 916:937 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 4 41 theme small 845:849 arg1 subunit 851:857 the small subunit 841:857 the small subunit 841:857 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 6 42 theme substrate 1256:1264 arg1 specificity 1266:1276 the substrate specificity 1252:1276 the substrate specificity of this unusual enzyme 1252:1299 Proper orientation of the two subunits thus seems very important for the substrate specificity of this unusual enzyme. 8089145 5 43 theme proteolytic 1010:1020 arg1 cleavage 1022:1029 proteolytic cleavage 1010:1029 proteolytic cleavage of the precursor protein 1010:1054 Remarkably, proteolytic cleavage of the precursor protein increases the activity of the enzyme toward LPS by 10-20-fold without altering its activity toward glycerophosphatidylcholine. 8089145 1 44 theme Acyloxyacyl 131:141 arg1 hydrolase 143:151 Acyloxyacyl hydrolase 131:151 Acyloxyacyl hydrolase 131:151 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 1 44 theme Acyloxyacyl 131:141 arg1 polypeptide 277:287 a precursor polypeptide 265:287 a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits 265:386 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 1 44 theme Acyloxyacyl 131:141 arg1 enzyme 166:171 a leukocyte enzyme 154:171 a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids 154:244 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 2 45 theme sequence 550:557 arg1 similarity 559:568 amino acid sequence similarity 539:568 amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases 539:658 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 4 46 from processing 928:937 arg1 cells 991:995 baby hamster kidney cells 971:995 baby hamster kidney cells 971:995 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 4 47 contain have 859:862 arg1 mutations 807:815 mutations 807:815 mutations that truncate or delete the small subunit 807:857 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 4 47 contain have 859:862 arg2 effects 873:879 profound effects 864:879 profound effects 864:879 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 8089145 1 48 theme many 227:230 arg1 glycerolipids 232:244 many glycerolipids 227:244 many glycerolipids 227:244 Acyloxyacyl hydrolase, a leukocyte enzyme that acts on bacterial lipopolysaccharides (LPSs) and many glycerolipids, is synthesized as a precursor polypeptide that undergoes internal disulfide linkage before being proteolytically processed into two subunits. 8089145 2 49 theme lipases 500:506 arg1 sites 486:490 the active sites 475:490 the active sites of many lipases 475:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 2 49 theme lipases 500:506 arg1 lipases 500:506 many lipases 495:506 many lipases 495:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 5 50 theme enzyme 1086:1091 arg1 activity 1070:1077 the activity 1066:1077 the activity of the enzyme toward LPS 1066:1102 Remarkably, proteolytic cleavage of the precursor protein increases the activity of the enzyme toward LPS by 10-20-fold without altering its activity toward glycerophosphatidylcholine. 8089145 2 51 theme many 495:498 arg1 lipases 500:506 many lipases 495:506 many lipases 495:506 The larger subunit contains an amino acid sequence (Gly-X-Ser-X-Gly) that is found at the active sites of many lipases, while the smaller subunit has amino acid sequence similarity to saposins (sphingolipid activator proteins), cofactors for sphingolipid glycohydrolases. 8089145 4 52 theme hamster 976:982 arg1 kidney 984:989 baby hamster kidney 971:989 baby hamster kidney cells 971:995 In addition, mutations that truncate or delete the small subunit have profound effects on the intracellular localization, proteolytic processing, and stability of the enzyme in baby hamster kidney cells. 11866528 10 0 theme Aplysia 1500:1506 arg1 cyclase 1508:1514 Aplysia cyclase 1500:1514 Aplysia cyclase 1500:1514 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 4 1 theme structural 480:489 arg1 basis 491:495 the structural basis 476:495 the structural basis of these enzymatic reactions 476:524 In spite of its functional importance, the structural basis of these enzymatic reactions remains elusive. 11866528 0 2 with BST-1/CD157 34:44 arg1 activities 94:103 NAD glycohydrolase activities 75:103 NAD glycohydrolase activities 75:103 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 0 2 with BST-1/CD157 34:44 arg1 cyclase 63:69 ADP-ribosyl cyclase 51:69 ADP-ribosyl cyclase 51:69 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 9 3 theme adenine 1394:1400 arg1 base 1402:1405 the adenine base and the ribose 1390:1420 base 1402:1405 This conformation of the catalytic cleft also implies the mechanism of cyclization between the adenine base and the ribose. 11866528 0 4 theme glycohydrolase 79:92 arg1 activities 94:103 NAD glycohydrolase activities 75:103 NAD glycohydrolase activities 75:103 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 0 5 from studies 17:23 arg1 BST-1/CD157 34:44 human BST-1/CD157 28:44 human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities 28:103 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 5 6 theme extracellular 587:599 arg1 region 601:606 the extracellular region 583:606 the extracellular region of human BST-1 583:621 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 7 7 theme glutamate 1189:1197 arg1 residue 1199:1205 a catalytic glutamate residue 1177:1205 a catalytic glutamate residue 1177:1205 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 10 8 theme BST-1 1483:1487 arg1 sequences 1470:1478 the sequences 1466:1478 the sequences of BST-1, CD38, and Aplysia cyclase 1466:1514 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 5 9 theme free 651:654 arg1 form 656:659 the free form 647:659 the free form 647:659 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 3 10 contain bear 381:384 arg2 activities 425:434 NAD glycohydrolase activities 406:434 NAD glycohydrolase activities 406:434 These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. 11866528 3 10 contain bear 381:384 arg2 hydrolase 392:400 cADPR hydrolase 386:400 cADPR hydrolase 386:400 These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. 11866528 3 10 contain bear 381:384 arg1 enzymes 347:353 These enzymes 341:353 These enzymes 341:353 These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. 11866528 10 11 theme substrate 1532:1540 arg1 mode 1554:1557 this substrate recognition mode 1527:1557 this substrate recognition mode 1527:1557 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 6 12 theme binding 878:884 arg1 mode 860:863 the mode 856:863 the mode of substrate binding 856:884 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 6 12 theme binding 878:884 arg1 mechanism 904:912 the catalytic mechanism 890:912 the catalytic mechanism of the multifunctional enzymatic reactions 890:955 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 6 13 theme structural 788:797 arg1 views 799:803 The three-dimensional structural views 766:803 The three-dimensional structural views of the reaction centre with these ligands 766:845 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 8 14 theme N 1279:1279 arg1 reaction 1289:1296 the S(N)-1 type reaction 1273:1296 the S(N)-1 type reaction 1273:1296 Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. 11866528 8 15 theme carboxyl 1212:1219 arg1 side-chain 1221:1230 Its carboxyl side-chain 1208:1230 Its carboxyl side-chain 1208:1230 Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. 11866528 4 16 theme functional 453:462 arg1 importance 464:473 its functional importance 449:473 its functional importance 449:473 In spite of its functional importance, the structural basis of these enzymatic reactions remains elusive. 11866528 5 17 theme BST-1 617:621 arg1 region 601:606 the extracellular region 583:606 the extracellular region of human BST-1 583:621 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 18 theme human 611:615 arg1 BST-1 617:621 human BST-1 611:621 human BST-1 611:621 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 2 19 theme cyclase 267:273 arg1 function 275:282 the ADP-ribosyl cyclase function 251:282 the ADP-ribosyl cyclase function 251:282 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 1 20 theme intracellular 176:188 arg1 stores 198:203 intracellular calcium stores 176:203 intracellular calcium stores 176:203 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 0 21 theme ADP-ribosyl 51:61 arg1 cyclase 63:69 ADP-ribosyl cyclase 51:69 ADP-ribosyl cyclase 51:69 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 5 22 theme crystal 561:567 arg1 structures 569:578 the crystal structures 557:578 the crystal structures of the extracellular region of human BST-1 557:621 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 2 23 theme ADP-ribosyl 255:265 arg1 cyclase 267:273 ADP-ribosyl cyclase 255:273 the ADP-ribosyl cyclase function 251:282 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 1 24 theme calcium 190:196 arg1 stores 198:203 intracellular calcium stores 176:203 intracellular calcium stores 176:203 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 10 25 theme catalytic 1563:1571 arg1 scheme 1573:1578 catalytic scheme 1563:1578 catalytic scheme 1563:1578 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 9 26 theme cleft 1334:1338 arg1 conformation 1304:1315 This conformation 1299:1315 This conformation of the catalytic cleft 1299:1338 This conformation of the catalytic cleft also implies the mechanism of cyclization between the adenine base and the ribose. 11866528 5 27 theme atomic 626:631 arg1 resolution 633:642 atomic resolution 626:642 atomic resolution in the free form 626:659 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 28 dep analogues 698:706 arg1 analogues 698:706 five substrate analogues 683:706 five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD 683:763 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 28 dep analogues 698:706 arg1 ATPgammaS 728:736 ATPgammaS 728:736 ATPgammaS 728:736 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 28 dep analogues 698:706 arg1 ethenoNADP 739:748 ethenoNADP 739:748 ethenoNADP 739:748 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 28 dep analogues 698:706 arg1 nicotinamide 709:720 nicotinamide 709:720 nicotinamide 709:720 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 28 dep analogues 698:706 arg1 NMN 723:725 NMN 723:725 NMN 723:725 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 28 dep analogues 698:706 arg1 ethenoNAD 755:763 ethenoNAD 755:763 ethenoNAD 755:763 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 2 29 theme membrane 300:307 arg1 CD38 319:322 CD38 319:322 CD38 319:322 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 2 29 theme membrane 300:307 arg1 proteins 309:316 two membrane proteins 296:316 two membrane proteins 296:316 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 2 29 theme membrane 300:307 arg1 BST-1/CD157 328:338 BST-1/CD157 328:338 BST-1/CD157 328:338 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 6 30 theme substrate 868:876 arg1 binding 878:884 substrate binding 868:884 substrate binding 868:884 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 3 31 theme NAD 406:408 arg1 activities 425:434 NAD glycohydrolase activities 406:434 NAD glycohydrolase activities 406:434 These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. 11866528 0 32 theme Crystallographic 0:15 arg1 studies 17:23 Crystallographic studies 0:23 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 0:104 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 7 33 theme NAD 1147:1149 arg1 bond 1139:1142 the N-glycosidic bond 1122:1142 the N-glycosidic bond of NAD 1122:1149 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 10 34 theme key 1433:1435 arg1 residues 1437:1444 The three key residues 1423:1444 The three key residues 1423:1444 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 10 34 theme key 1433:1435 arg1 invariant 1450:1458 invariant 1450:1458 invariant 1450:1458 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 7 35 theme tryptophan 1089:1098 arg1 residues 1100:1107 two tryptophan residues 1085:1107 two tryptophan residues 1085:1107 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 3 36 theme glycohydrolase 410:423 arg1 activities 425:434 NAD glycohydrolase activities 406:434 NAD glycohydrolase activities 406:434 These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. 11866528 0 37 theme human 28:32 arg1 BST-1/CD157 34:44 human BST-1/CD157 28:44 human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities 28:103 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 5 38 from resolution 633:642 arg1 form 656:659 the free form 647:659 the free form 647:659 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 7 39 theme Waals 1061:1065 arg1 interactions 1067:1078 van der Waals interactions 1053:1078 van der Waals interactions with two tryptophan residues 1053:1107 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 6 40 theme three-dimensional 770:786 arg1 views 799:803 The three-dimensional structural views 766:803 The three-dimensional structural views of the reaction centre with these ligands 766:845 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 6 41 with views 799:803 arg1 ligands 839:845 these ligands 833:845 these ligands 833:845 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 2 42 located found 287:291 arg1 CD38 319:322 CD38 319:322 CD38 319:322 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 2 42 located found 287:291 arg1 mammals 242:248 mammals 242:248 mammals 242:248 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 2 42 located found 287:291 arg1 proteins 309:316 two membrane proteins 296:316 two membrane proteins 296:316 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 2 42 located found 287:291 arg1 BST-1/CD157 328:338 BST-1/CD157 328:338 BST-1/CD157 328:338 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 2 42 located found 287:291 arg2 function 275:282 the ADP-ribosyl cyclase function 251:282 the ADP-ribosyl cyclase function 251:282 In mammals, the ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and BST-1/CD157. 11866528 5 43 theme substrate 688:696 arg1 analogues 698:706 five substrate analogues 683:706 five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD 683:763 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 43 theme substrate 688:696 arg1 ATPgammaS 728:736 ATPgammaS 728:736 ATPgammaS 728:736 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 43 theme substrate 688:696 arg1 ethenoNADP 739:748 ethenoNADP 739:748 ethenoNADP 739:748 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 43 theme substrate 688:696 arg1 nicotinamide 709:720 nicotinamide 709:720 nicotinamide 709:720 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 43 theme substrate 688:696 arg1 NMN 723:725 NMN 723:725 NMN 723:725 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 43 theme substrate 688:696 arg1 ethenoNAD 755:763 ethenoNAD 755:763 ethenoNAD 755:763 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 6 44 theme catalytic 894:902 arg1 mechanism 904:912 the catalytic mechanism 890:912 the catalytic mechanism of the multifunctional enzymatic reactions 890:955 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 6 45 theme reactions 947:955 arg1 mode 860:863 the mode 856:863 the mode of substrate binding 856:884 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 6 45 theme reactions 947:955 arg1 mechanism 904:912 the catalytic mechanism 890:912 the catalytic mechanism of the multifunctional enzymatic reactions 890:955 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 7 46 theme van 1053:1055 arg1 interactions 1067:1078 van der Waals interactions 1053:1078 van der Waals interactions with two tryptophan residues 1053:1107 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 8 47 theme catalytic 1247:1255 arg1 intermediate 1257:1268 the catalytic intermediate 1243:1268 the catalytic intermediate of the S(N)-1 type reaction 1243:1296 Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. 11866528 9 48 theme catalytic 1324:1332 arg1 cleft 1334:1338 the catalytic cleft 1320:1338 the catalytic cleft 1320:1338 This conformation of the catalytic cleft also implies the mechanism of cyclization between the adenine base and the ribose. 11866528 5 49 with complexes 668:676 arg1 analogues 698:706 five substrate analogues 683:706 five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD 683:763 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 49 with complexes 668:676 arg1 ATPgammaS 728:736 ATPgammaS 728:736 ATPgammaS 728:736 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 49 with complexes 668:676 arg1 ethenoNADP 739:748 ethenoNADP 739:748 ethenoNADP 739:748 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 49 with complexes 668:676 arg1 nicotinamide 709:720 nicotinamide 709:720 nicotinamide 709:720 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 49 with complexes 668:676 arg1 NMN 723:725 NMN 723:725 NMN 723:725 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 5 49 with complexes 668:676 arg1 ethenoNAD 755:763 ethenoNAD 755:763 ethenoNAD 755:763 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 4 50 theme reactions 516:524 arg1 basis 491:495 the structural basis 476:495 the structural basis of these enzymatic reactions 476:524 In spite of its functional importance, the structural basis of these enzymatic reactions remains elusive. 11866528 7 51 theme der 1057:1059 arg1 interactions 1067:1078 van der Waals interactions 1053:1078 van der Waals interactions with two tryptophan residues 1053:1107 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 7 52 theme catalytic 1179:1187 arg1 residue 1199:1205 a catalytic glutamate residue 1177:1205 a catalytic glutamate residue 1177:1205 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 7 53 theme N-glycosidic 1126:1137 arg1 bond 1139:1142 the N-glycosidic bond 1122:1142 the N-glycosidic bond of NAD 1122:1149 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 4 54 theme enzymatic 506:514 arg1 reactions 516:524 these enzymatic reactions 500:524 these enzymatic reactions 500:524 In spite of its functional importance, the structural basis of these enzymatic reactions remains elusive. 11866528 10 55 from common 1593:1598 arg1 family 1615:1620 the cyclase family 1603:1620 the cyclase family 1603:1620 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 1 56 theme novel 119:123 arg1 messenger 132:140 the novel second messenger 115:140 the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3) 115:236 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 1 56 theme novel 119:123 arg1 cADPR 106:110 cADPR 106:110 cADPR 106:110 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 3 57 theme cADPR 386:390 arg1 hydrolase 392:400 cADPR hydrolase 386:400 cADPR hydrolase 386:400 These enzymes, exposed extracellularly, bear cADPR hydrolase and NAD glycohydrolase activities. 11866528 10 58 theme CD38 1490:1493 arg1 sequences 1470:1478 the sequences 1466:1478 the sequences of BST-1, CD38, and Aplysia cyclase 1466:1514 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 7 59 theme dimeric 989:995 arg1 enzyme 997:1002 the dimeric enzyme 985:1002 the dimeric enzyme 985:1002 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 1 60 theme second 125:130 arg1 messenger 132:140 the novel second messenger 115:140 the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3) 115:236 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 1 60 theme second 125:130 arg1 cADPR 106:110 cADPR 106:110 cADPR 106:110 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 6 61 theme enzymatic 937:945 arg1 reactions 947:955 the multifunctional enzymatic reactions 917:955 the multifunctional enzymatic reactions 917:955 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 6 62 theme centre 821:826 arg1 views 799:803 The three-dimensional structural views 766:803 The three-dimensional structural views of the reaction centre with these ligands 766:845 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 5 63 theme region 601:606 arg1 structures 569:578 the crystal structures 557:578 the crystal structures of the extracellular region of human BST-1 557:621 We determined the crystal structures of the extracellular region of human BST-1 at atomic resolution in the free form and in complexes with five substrate analogues: nicotinamide, NMN, ATPgammaS, ethenoNADP, and ethenoNAD. 11866528 8 64 theme type 1284:1287 arg1 reaction 1289:1296 the S(N)-1 type reaction 1273:1296 the S(N)-1 type reaction 1273:1296 Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. 11866528 8 65 theme S 1277:1277 arg1 reaction 1289:1296 the S(N)-1 type reaction 1273:1296 the S(N)-1 type reaction 1273:1296 Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. 11866528 7 66 theme enzyme 997:1002 arg1 cleft 976:980 each catalytic cleft 961:980 each catalytic cleft of the dimeric enzyme 961:1002 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 6 67 theme reaction 812:819 arg1 centre 821:826 the reaction centre 808:826 the reaction centre 808:826 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 11866528 1 68 from stores 198:203 arg1 release 163:169 calcium release 155:169 calcium release from intracellular calcium stores 155:203 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 8 69 theme reaction 1289:1296 arg1 intermediate 1257:1268 the catalytic intermediate 1243:1268 the catalytic intermediate of the S(N)-1 type reaction 1243:1296 Its carboxyl side-chain stabilizes the catalytic intermediate of the S(N)-1 type reaction. 11866528 9 70 theme cyclization 1370:1380 arg1 mechanism 1357:1365 the mechanism 1353:1365 the mechanism of cyclization between the adenine base and the ribose 1353:1420 This conformation of the catalytic cleft also implies the mechanism of cyclization between the adenine base and the ribose. 11866528 0 71 theme NAD 75:77 arg1 activities 94:103 NAD glycohydrolase activities 75:103 NAD glycohydrolase activities 75:103 Crystallographic studies on human BST-1/CD157 with ADP-ribosyl cyclase and NAD glycohydrolase activities. 11866528 7 72 with interactions 1067:1078 arg1 residues 1100:1107 two tryptophan residues 1085:1107 two tryptophan residues 1085:1107 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 10 73 from family 1615:1620 arg1 common 1593:1598 common 1593:1598 common 1593:1598 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 10 74 theme cyclase 1607:1613 arg1 family 1615:1620 the cyclase family 1603:1620 the cyclase family 1603:1620 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 10 75 theme cyclase 1508:1514 arg1 sequences 1470:1478 the sequences 1466:1478 the sequences of BST-1, CD38, and Aplysia cyclase 1466:1514 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 10 76 theme recognition 1542:1552 arg1 mode 1554:1557 this substrate recognition mode 1527:1557 this substrate recognition mode 1527:1557 The three key residues are invariant among the sequences of BST-1, CD38, and Aplysia cyclase, and hence this substrate recognition mode and catalytic scheme appear to be common in the cyclase family. 11866528 7 77 theme catalytic 966:974 arg1 cleft 976:980 each catalytic cleft 961:980 each catalytic cleft of the dimeric enzyme 961:1002 In each catalytic cleft of the dimeric enzyme, substrates are recognized predominantly through van der Waals interactions with two tryptophan residues, and thereby the N-glycosidic bond of NAD is correctly exposed near a catalytic glutamate residue. 11866528 1 78 theme calcium 155:161 arg1 release 163:169 calcium release 155:169 calcium release from intracellular calcium stores 155:203 cADPR is the novel second messenger that elicits calcium release from intracellular calcium stores and works independently of IP(3). 11866528 6 79 theme multifunctional 921:935 arg1 reactions 947:955 the multifunctional enzymatic reactions 917:955 the multifunctional enzymatic reactions 917:955 The three-dimensional structural views of the reaction centre with these ligands revealed the mode of substrate binding and the catalytic mechanism of the multifunctional enzymatic reactions. 15099525 4 0 from existence 568:576 arg1 membrane 621:628 the cell membrane 612:628 the cell membrane 612:628 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 4 1 theme dimerization 450:461 arg1 interaction 463:473 The observed intimate dimerization interaction 428:473 The observed intimate dimerization interaction at D4 428:479 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 4 2 from D4 478:479 arg1 interaction 463:473 The observed intimate dimerization interaction 428:473 The observed intimate dimerization interaction at D4 428:479 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 4 2 from D4 478:479 arg1 architecture 509:520 a stiff D4-D5 stem-like architecture 485:520 a stiff D4-D5 stem-like architecture 485:520 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 1 3 theme C-terminal 125:134 arg1 ICAM-1 159:164 ICAM-1 159:164 ICAM-1 159:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 1 3 theme C-terminal 125:134 arg1 domains 136:142 the three C-terminal domains 3-5 (D3-D5) 115:154 the three C-terminal domains 3-5 (D3-D5) of ICAM-1 115:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 4 4 theme structural 537:546 arg1 explanation 548:558 a good structural explanation 530:558 a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane 530:628 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 5 5 theme immunological 839:851 arg1 synapse 853:859 an immunological synapse 836:859 an immunological synapse 836:859 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 1 6 theme domains 136:142 arg1 structure 102:110 the 3.0 A crystal structure 84:110 the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1 84:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 4 7 theme good 532:535 arg1 explanation 548:558 a good structural explanation 530:558 a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane 530:628 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 4 8 theme cell 616:619 arg1 membrane 621:628 the cell membrane 612:628 the cell membrane 612:628 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 5 9 theme band-like 685:693 arg1 cluster 718:724 a band-like one-dimensional linear cluster 683:724 a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface 683:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 2 10 with Combined 167:174 arg1 D1D2 235:238 D1D2 235:238 D1D2 235:238 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 2 10 with Combined 167:174 arg1 structure 224:232 the previously known N-terminal two-domain structure 181:232 the previously known N-terminal two-domain structure (D1D2) 181:239 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 5 11 theme T 882:882 arg1 critical 906:913 critical 906:913 critical 906:913 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 11 theme T 882:882 arg1 cell 884:887 an activated T cell 869:887 an activated T cell 869:887 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 12 theme one-dimensional 695:709 arg1 cluster 718:724 a band-like one-dimensional linear cluster 683:724 a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface 683:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 4 13 theme ICAM-1 591:596 arg1 dimers 602:607 preformed ICAM-1 cis dimers 581:607 preformed ICAM-1 cis dimers 581:607 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 3 14 theme members 387:393 arg1 ICAM-3 409:414 ICAM-3 409:414 ICAM-3 409:414 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 3 14 theme members 387:393 arg1 architecture 353:364 a general architecture 343:364 a general architecture of other ICAM family members 343:393 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 3 14 theme members 387:393 arg1 ICAM-5 420:425 ICAM-5 420:425 ICAM-5 420:425 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 5 15 theme linear 711:716 arg1 cluster 718:724 a band-like one-dimensional linear cluster 683:724 a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface 683:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 4 16 theme intimate 441:448 arg1 interaction 463:473 The observed intimate dimerization interaction 428:473 The observed intimate dimerization interaction at D4 428:479 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 5 17 from D1 679:680 arg1 interface 666:674 another dimerization interface 645:674 another dimerization interface at D1 645:680 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 18 theme T 919:919 arg1 signaling 935:943 T cell receptor signaling 919:943 T cell receptor signaling 919:943 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 4 19 theme observed 432:439 arg1 interaction 463:473 The observed intimate dimerization interaction 428:473 The observed intimate dimerization interaction at D4 428:479 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 2 20 theme entire 256:261 arg1 fragment 284:291 an entire ICAM-1 extracellular fragment 253:291 an entire ICAM-1 extracellular fragment 253:291 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 3 21 theme ICAM 375:378 arg1 members 387:393 other ICAM family members 369:393 other ICAM family members 369:393 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 0 22 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for dimerization of ICAM-1 on the cell surface. 0:63 Structural basis for dimerization of ICAM-1 on the cell surface. 15099525 1 23 dep domains 136:142 arg1 D3-D5 149:153 D3-D5 149:153 D3-D5 149:153 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 1 23 dep domains 136:142 arg1 3-5 144:146 3-5 144:146 3-5 144:146 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 3 24 theme family 380:385 arg1 members 387:393 other ICAM family members 369:393 other ICAM family members 369:393 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 4 25 theme stem-like 499:507 arg1 architecture 509:520 a stiff D4-D5 stem-like architecture 485:520 a stiff D4-D5 stem-like architecture 485:520 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 5 26 theme cell 761:764 arg1 surface 772:778 an antigen-presenting cell (APC) surface 739:778 an antigen-presenting cell (APC) surface 739:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 4 27 theme cis 598:600 arg1 dimers 602:607 preformed ICAM-1 cis dimers 581:607 preformed ICAM-1 cis dimers 581:607 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 5 28 theme synapse 853:859 arg1 formation 823:831 the formation 819:831 the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling 819:943 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 1 29 theme ICAM-1 159:164 arg1 ICAM-1 159:164 ICAM-1 159:164 ICAM-1 159:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 1 29 theme ICAM-1 159:164 arg1 domains 136:142 the three C-terminal domains 3-5 (D3-D5) 115:154 the three C-terminal domains 3-5 (D3-D5) of ICAM-1 115:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 0 30 theme ICAM-1 37:42 arg1 dimerization 21:32 dimerization 21:32 dimerization of ICAM-1 on the cell surface 21:62 Structural basis for dimerization of ICAM-1 on the cell surface. 15099525 2 31 theme fragment 284:291 arg1 model 244:248 a model 242:248 a model of an entire ICAM-1 extracellular fragment 242:291 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 2 32 theme known 196:200 arg1 D1D2 235:238 D1D2 235:238 D1D2 235:238 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 2 32 theme known 196:200 arg1 structure 224:232 the previously known N-terminal two-domain structure 181:232 the previously known N-terminal two-domain structure (D1D2) 181:239 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 3 33 theme other 369:373 arg1 members 387:393 other ICAM family members 369:393 other ICAM family members 369:393 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 5 34 theme antigen-presenting 742:759 arg1 cell 761:764 antigen-presenting cell 742:764 an antigen-presenting cell (APC) surface 739:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 34 theme antigen-presenting 742:759 arg1 APC 767:769 APC 767:769 APC 767:769 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 4 35 theme dimers 602:607 arg1 existence 568:576 the existence 564:576 the existence of preformed ICAM-1 cis dimers on the cell membrane 564:628 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 3 36 theme general 345:351 arg1 ICAM-3 409:414 ICAM-3 409:414 ICAM-3 409:414 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 3 36 theme general 345:351 arg1 architecture 353:364 a general architecture 343:364 a general architecture of other ICAM family members 343:393 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 3 36 theme general 345:351 arg1 ICAM-5 420:425 ICAM-5 420:425 ICAM-5 420:425 This model should represent a general architecture of other ICAM family members, particularly ICAM-3 and ICAM-5. 15099525 5 37 theme ICAM-1 729:734 arg1 cluster 718:724 a band-like one-dimensional linear cluster 683:724 a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface 683:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 1 38 theme A 92:92 arg1 structure 102:110 the 3.0 A crystal structure 84:110 the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1 84:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 4 39 theme D4-D5 493:497 arg1 architecture 509:520 a stiff D4-D5 stem-like architecture 485:520 a stiff D4-D5 stem-like architecture 485:520 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 1 40 theme crystal 94:100 arg1 structure 102:110 the 3.0 A crystal structure 84:110 the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1 84:164 We have determined the 3.0 A crystal structure of the three C-terminal domains 3-5 (D3-D5) of ICAM-1. 15099525 4 41 theme stiff 487:491 arg1 architecture 509:520 a stiff D4-D5 stem-like architecture 485:520 a stiff D4-D5 stem-like architecture 485:520 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 5 42 theme cell 921:924 arg1 signaling 935:943 T cell receptor signaling 919:943 T cell receptor signaling 919:943 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 2 43 theme extracellular 270:282 arg1 fragment 284:291 an entire ICAM-1 extracellular fragment 253:291 an entire ICAM-1 extracellular fragment 253:291 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 5 44 theme dimerization 653:664 arg1 interface 666:674 another dimerization interface 645:674 another dimerization interface at D1 645:680 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 45 from cluster 718:724 arg1 surface 772:778 an antigen-presenting cell (APC) surface 739:778 an antigen-presenting cell (APC) surface 739:778 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 46 theme receptor 926:933 arg1 signaling 935:943 T cell receptor signaling 919:943 T cell receptor signaling 919:943 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 2 47 theme ICAM-1 263:268 arg1 fragment 284:291 an entire ICAM-1 extracellular fragment 253:291 an entire ICAM-1 extracellular fragment 253:291 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 5 48 theme activated 872:880 arg1 critical 906:913 critical 906:913 critical 906:913 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 5 48 theme activated 872:880 arg1 cell 884:887 an activated T cell 869:887 an activated T cell 869:887 Together with another dimerization interface at D1, a band-like one-dimensional linear cluster of ICAM-1 on an antigen-presenting cell (APC) surface can be envisioned, which might explain the formation of an immunological synapse between an activated T cell and APC which is critical for T cell receptor signaling. 15099525 2 49 theme two-domain 213:222 arg1 D1D2 235:238 D1D2 235:238 D1D2 235:238 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 2 49 theme two-domain 213:222 arg1 structure 224:232 the previously known N-terminal two-domain structure 181:232 the previously known N-terminal two-domain structure (D1D2) 181:239 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 4 50 theme preformed 581:589 arg1 dimers 602:607 preformed ICAM-1 cis dimers 581:607 preformed ICAM-1 cis dimers 581:607 The observed intimate dimerization interaction at D4 and a stiff D4-D5 stem-like architecture provide a good structural explanation for the existence of preformed ICAM-1 cis dimers on the cell membrane. 15099525 0 51 theme cell 51:54 arg1 surface 56:62 the cell surface 47:62 the cell surface 47:62 Structural basis for dimerization of ICAM-1 on the cell surface. 15099525 0 52 from dimerization 21:32 arg1 surface 56:62 the cell surface 47:62 the cell surface 47:62 Structural basis for dimerization of ICAM-1 on the cell surface. 15099525 2 53 theme N-terminal 202:211 arg1 D1D2 235:238 D1D2 235:238 D1D2 235:238 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 15099525 2 53 theme N-terminal 202:211 arg1 structure 224:232 the previously known N-terminal two-domain structure 181:232 the previously known N-terminal two-domain structure (D1D2) 181:239 Combined with the previously known N-terminal two-domain structure (D1D2), a model of an entire ICAM-1 extracellular fragment has been constructed. 19520913 7 0 theme uptake 1123:1128 arg1 regulation 1097:1106 sterol regulation 1090:1106 sterol regulation of cholesterol uptake 1090:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 4 1 theme LXR 668:670 arg1 ligand 672:677 LXR ligand 668:677 LXR ligand 668:677 LXR ligand reduces, whereas LXR knockout increases, LDLR protein levels in vivo in a tissue-selective manner. 19520913 0 2 theme receptor 82:89 arg1 ubiquitination 56:69 Idol-dependent ubiquitination 41:69 Idol-dependent ubiquitination of the LDL receptor 41:89 LXR regulates cholesterol uptake through Idol-dependent ubiquitination of the LDL receptor. 19520913 6 3 theme adenovirus-mediated 875:893 arg1 expression 895:904 adenovirus-mediated expression 875:904 adenovirus-mediated expression of Idol in mouse liver 875:927 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 3 4 theme Idol 500:503 arg1 induction 487:495 transcriptional induction 471:495 transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation 471:665 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 6 5 theme LDL 975:977 arg1 levels 979:984 plasma LDL levels 968:984 plasma LDL levels 968:984 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 2 6 theme nuclear 228:234 arg1 LXR 254:256 LXR 254:256 LXR 254:256 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 6 theme nuclear 228:234 arg1 receptor 244:251 the sterol-responsive nuclear liver X receptor 206:251 the sterol-responsive nuclear liver X receptor (LXR) 206:257 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 4 7 theme protein 725:731 arg1 levels 733:738 LDLR protein levels 720:738 LDLR protein levels 720:738 LXR ligand reduces, whereas LXR knockout increases, LDLR protein levels in vivo in a tissue-selective manner. 19520913 6 8 theme plasma 968:973 arg1 levels 979:984 plasma LDL levels 968:984 plasma LDL levels 968:984 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 4 9 theme knockout 700:707 arg1 increases 709:717 LXR knockout increases 696:717 LXR knockout increases 696:717 LXR ligand reduces, whereas LXR knockout increases, LDLR protein levels in vivo in a tissue-selective manner. 19520913 3 10 theme LDLR 532:535 arg1 degrader 516:523 inducible degrader 506:523 inducible degrader of the LDLR 506:535 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 2 11 theme cholesterol 274:284 arg1 homeostasis 286:296 cholesterol homeostasis 274:296 cholesterol homeostasis 274:296 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 3 12 from ubiquitination 576:589 arg1 domain 622:627 its cytoplasmic domain 606:627 its cytoplasmic domain 606:627 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 3 13 theme transcriptional 471:485 arg1 induction 487:495 transcriptional induction 471:495 transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation 471:665 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 7 14 theme sterol 1090:1095 arg1 regulation 1097:1106 sterol regulation 1090:1106 sterol regulation of cholesterol uptake 1090:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 1 15 theme endogenous 166:175 arg1 synthesis 177:185 endogenous synthesis 166:185 endogenous synthesis 166:185 Cellular cholesterol levels reflect a balance between uptake, efflux, and endogenous synthesis. 19520913 3 16 from domain 622:627 arg1 ubiquitination 576:589 ubiquitination 576:589 ubiquitination of the LDLR on its cytoplasmic domain 576:627 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 2 17 theme lipoprotein 392:402 arg1 uptake 410:415 low-density lipoprotein (LDL) uptake 380:415 low-density lipoprotein (LDL) uptake 380:415 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 6 18 from expression 895:904 arg1 liver 923:927 mouse liver 917:927 mouse liver 917:927 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 1 19 theme Cellular 92:99 arg1 levels 113:118 Cellular cholesterol levels 92:118 Cellular cholesterol levels 92:118 Cellular cholesterol levels reflect a balance between uptake, efflux, and endogenous synthesis. 19520913 3 20 from LDLR 598:601 arg1 domain 622:627 its cytoplasmic domain 606:627 its cytoplasmic domain 606:627 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 2 21 theme low-density 380:390 arg1 lipoprotein 392:402 low-density lipoprotein 380:402 low-density lipoprotein (LDL) uptake 380:415 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 21 theme low-density 380:390 arg1 LDL 405:407 LDL 405:407 LDL 405:407 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 1 22 theme cholesterol 101:111 arg1 levels 113:118 Cellular cholesterol levels 92:118 Cellular cholesterol levels 92:118 Cellular cholesterol levels reflect a balance between uptake, efflux, and endogenous synthesis. 19520913 3 23 dep Idol 500:503 arg1 degrader 516:523 inducible degrader 506:523 inducible degrader of the LDLR 506:535 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 5 24 theme Idol 778:781 arg1 knockdown 783:791 Idol knockdown 778:791 Idol knockdown in hepatocytes 778:806 Idol knockdown in hepatocytes increases LDLR protein levels and promotes LDL uptake. 19520913 4 25 theme LXR 696:698 arg1 increases 709:717 LXR knockout increases 696:717 LXR knockout increases 696:717 LXR ligand reduces, whereas LXR knockout increases, LDLR protein levels in vivo in a tissue-selective manner. 19520913 7 26 theme response 1052:1059 arg1 proteins 1077:1084 sterol response element-binding proteins 1045:1084 sterol response element-binding proteins for sterol regulation of cholesterol uptake 1045:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 3 27 theme E3 542:543 arg1 Idol 500:503 Idol 500:503 Idol (inducible degrader of the LDLR) 500:536 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 3 27 theme E3 542:543 arg1 ligase 555:560 an E3 ubiquitin ligase 539:560 an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation 539:665 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 6 28 theme LDLR 938:941 arg1 degradation 943:953 LDLR degradation 938:953 LDLR degradation 938:953 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 7 29 theme element-binding 1061:1075 arg1 proteins 1077:1084 sterol response element-binding proteins 1045:1084 sterol response element-binding proteins for sterol regulation of cholesterol uptake 1045:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 3 30 theme cytoplasmic 610:620 arg1 domain 622:627 its cytoplasmic domain 606:627 its cytoplasmic domain 606:627 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 2 31 theme uptake 410:415 arg1 suppression 365:375 suppression 365:375 suppression of low-density lipoprotein (LDL) uptake 365:415 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 0 32 theme Idol-dependent 41:54 arg1 ubiquitination 56:69 Idol-dependent ubiquitination 41:69 Idol-dependent ubiquitination of the LDL receptor 41:89 LXR regulates cholesterol uptake through Idol-dependent ubiquitination of the LDL receptor. 19520913 5 33 from knockdown 783:791 arg1 hepatocytes 796:806 hepatocytes 796:806 hepatocytes 796:806 Idol knockdown in hepatocytes increases LDLR protein levels and promotes LDL uptake. 19520913 2 34 theme X 242:242 arg1 LXR 254:256 LXR 254:256 LXR 254:256 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 34 theme X 242:242 arg1 receptor 244:251 the sterol-responsive nuclear liver X receptor 206:251 the sterol-responsive nuclear liver X receptor (LXR) 206:257 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 3 35 theme receptor 439:446 arg1 pathway 455:461 the LDL receptor (LDLR) pathway 431:461 the LDL receptor (LDLR) pathway 431:461 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 5 36 theme protein 823:829 arg1 levels 831:836 LDLR protein levels 818:836 LDLR protein levels 818:836 Idol knockdown in hepatocytes increases LDLR protein levels and promotes LDL uptake. 19520913 7 37 theme sterol 1045:1050 arg1 proteins 1077:1084 sterol response element-binding proteins 1045:1084 sterol response element-binding proteins for sterol regulation of cholesterol uptake 1045:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 4 38 theme LDLR 720:723 arg1 levels 733:738 LDLR protein levels 720:738 LDLR protein levels 720:738 LXR ligand reduces, whereas LXR knockout increases, LDLR protein levels in vivo in a tissue-selective manner. 19520913 2 39 theme liver 236:240 arg1 LXR 254:256 LXR 254:256 LXR 254:256 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 39 theme liver 236:240 arg1 receptor 244:251 the sterol-responsive nuclear liver X receptor 206:251 the sterol-responsive nuclear liver X receptor (LXR) 206:257 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 7 40 theme LXR-Idol-LDLR 991:1003 arg1 axis 1005:1008 The LXR-Idol-LDLR axis 987:1008 The LXR-Idol-LDLR axis 987:1008 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 5 41 theme LDLR 818:821 arg1 levels 831:836 LDLR protein levels 818:836 LDLR protein levels 818:836 Idol knockdown in hepatocytes increases LDLR protein levels and promotes LDL uptake. 19520913 6 42 theme mouse 917:921 arg1 liver 923:927 mouse liver 917:927 mouse liver 917:927 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 7 43 theme complementary 1020:1032 arg1 pathway 1034:1040 a complementary pathway 1018:1040 a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake 1018:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 19520913 3 44 theme ubiquitin 545:553 arg1 Idol 500:503 Idol 500:503 Idol (inducible degrader of the LDLR) 500:536 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 3 44 theme ubiquitin 545:553 arg1 ligase 555:560 an E3 ubiquitin ligase 539:560 an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation 539:665 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 2 45 theme efflux 341:346 arg1 promotion 316:324 promotion 316:324 promotion of cholesterol efflux 316:346 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 46 theme sterol-responsive 210:226 arg1 LXR 254:256 LXR 254:256 LXR 254:256 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 46 theme sterol-responsive 210:226 arg1 receptor 244:251 the sterol-responsive nuclear liver X receptor 206:251 the sterol-responsive nuclear liver X receptor (LXR) 206:257 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 2 47 theme cholesterol 329:339 arg1 efflux 341:346 cholesterol efflux 329:346 cholesterol efflux 329:346 Here we show that the sterol-responsive nuclear liver X receptor (LXR) helps maintain cholesterol homeostasis, not only through promotion of cholesterol efflux but also through suppression of low-density lipoprotein (LDL) uptake. 19520913 6 48 theme Idol 909:912 arg1 expression 895:904 adenovirus-mediated expression 875:904 adenovirus-mediated expression of Idol in mouse liver 875:927 Conversely, adenovirus-mediated expression of Idol in mouse liver promotes LDLR degradation and elevates plasma LDL levels. 19520913 3 49 theme LDLR 598:601 arg1 ubiquitination 576:589 ubiquitination 576:589 ubiquitination of the LDLR on its cytoplasmic domain 576:627 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 0 50 theme LDL 78:80 arg1 receptor 82:89 the LDL receptor 74:89 the LDL receptor 74:89 LXR regulates cholesterol uptake through Idol-dependent ubiquitination of the LDL receptor. 19520913 4 51 theme tissue-selective 753:768 arg1 manner 770:775 a tissue-selective manner 751:775 a tissue-selective manner 751:775 LXR ligand reduces, whereas LXR knockout increases, LDLR protein levels in vivo in a tissue-selective manner. 19520913 5 52 theme LDL 851:853 arg1 uptake 855:860 LDL uptake 851:860 LDL uptake 851:860 Idol knockdown in hepatocytes increases LDLR protein levels and promotes LDL uptake. 19520913 3 53 theme LDL 435:437 arg1 LDLR 449:452 LDLR 449:452 LDLR 449:452 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 3 53 theme LDL 435:437 arg1 receptor 439:446 LDL receptor 435:446 the LDL receptor (LDLR) pathway 431:461 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 0 54 theme cholesterol 14:24 arg1 uptake 26:31 cholesterol uptake 14:31 cholesterol uptake 14:31 LXR regulates cholesterol uptake through Idol-dependent ubiquitination of the LDL receptor. 19520913 3 55 theme inducible 506:514 arg1 degrader 516:523 inducible degrader 506:523 inducible degrader of the LDLR 506:535 LXR inhibits the LDL receptor (LDLR) pathway through transcriptional induction of Idol (inducible degrader of the LDLR), an E3 ubiquitin ligase that triggers ubiquitination of the LDLR on its cytoplasmic domain, thereby targeting it for degradation. 19520913 7 56 theme cholesterol 1111:1121 arg1 uptake 1123:1128 cholesterol uptake 1111:1128 cholesterol uptake 1111:1128 The LXR-Idol-LDLR axis defines a complementary pathway to sterol response element-binding proteins for sterol regulation of cholesterol uptake. 27995897 0 0 theme loss-of-function 86:101 arg1 mechanisms 103:112 distinct loss-of-function mechanisms 77:112 distinct loss-of-function mechanisms 77:112 Neurodegenerative disease mutations in TREM2 reveal a functional surface and distinct loss-of-function mechanisms. 27995897 0 1 from mutations 26:34 arg1 TREM2 39:43 TREM2 39:43 TREM2 39:43 Neurodegenerative disease mutations in TREM2 reveal a functional surface and distinct loss-of-function mechanisms. 27995897 6 2 theme functional 1003:1012 arg1 differences 1014:1024 structural and functional differences 988:1024 structural and functional differences among genetic variants of TREM2 988:1056 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 4 3 theme Biophysical 604:614 arg1 methods 629:635 Biophysical and cellular methods 604:635 Biophysical and cellular methods 604:635 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 0 4 theme distinct 77:84 arg1 mechanisms 103:112 distinct loss-of-function mechanisms 77:112 distinct loss-of-function mechanisms 77:112 Neurodegenerative disease mutations in TREM2 reveal a functional surface and distinct loss-of-function mechanisms. 27995897 6 5 with differences 1164:1174 arg1 approaches 1207:1216 patient-specific medicine approaches 1181:1216 patient-specific medicine approaches for neurodegenerative disorders 1181:1248 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 3 6 theme TREM2 375:379 arg1 structure 389:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 2 7 theme functional 311:320 arg1 studies 322:328 structural and functional studies 296:328 structural and functional studies of wild-type and variant proteins 296:362 To determine how TREM2 variants contribute to these diseases, we performed structural and functional studies of wild-type and variant proteins. 27995897 4 8 theme cellular 620:627 arg1 methods 629:635 Biophysical and cellular methods 604:635 Biophysical and cellular methods 604:635 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 3 9 theme crystal 381:387 arg1 structure 389:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 4 10 theme TREM2 798:802 arg1 ligand 804:809 a TREM2 ligand 796:809 a TREM2 ligand 796:809 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 3 11 theme distinct 568:575 arg1 effects 577:583 distinct effects 568:583 distinct effects 568:583 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 2 12 theme variant 347:353 arg1 proteins 355:362 wild-type and variant proteins 333:362 wild-type and variant proteins 333:362 To determine how TREM2 variants contribute to these diseases, we performed structural and functional studies of wild-type and variant proteins. 27995897 3 13 theme 3.1 369:371 arg1 structure 389:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 5 14 theme risk 842:845 arg1 variants 847:854 the Alzheimer's risk variants 826:854 the Alzheimer's risk variants 826:854 Additionally, the Alzheimer's risk variants appear to epitope map a functional surface on TREM2 that is unique within the larger TREM family. 27995897 2 15 theme structural 296:305 arg1 studies 322:328 structural and functional studies 296:328 structural and functional studies of wild-type and variant proteins 296:362 To determine how TREM2 variants contribute to these diseases, we performed structural and functional studies of wild-type and variant proteins. 27995897 4 16 theme TREM2 718:722 arg1 expression 732:741 folded TREM2 surface expression 711:741 folded TREM2 surface expression 711:741 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 3 17 theme Å 373:373 arg1 structure 389:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure 365:397 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 1 18 attach linked 175:180 arg2 variations 123:132 Genetic variations 115:132 Genetic variations in the myeloid immune receptor TREM2 115:169 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 1 18 attach linked 175:180 arg1 diseases 211:218 several neurodegenerative diseases 185:218 several neurodegenerative diseases 185:218 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 6 19 theme genetic 1032:1038 arg1 variants 1040:1047 genetic variants 1032:1047 genetic variants of TREM2 1032:1056 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 6 20 theme patient-specific 1181:1196 arg1 approaches 1207:1216 patient-specific medicine approaches 1181:1216 patient-specific medicine approaches for neurodegenerative disorders 1181:1248 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 6 21 theme TREM2 1052:1056 arg1 variants 1040:1047 genetic variants 1032:1047 genetic variants of TREM2 1032:1056 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 1 22 theme several 185:191 arg1 diseases 211:218 several neurodegenerative diseases 185:218 several neurodegenerative diseases 185:218 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 0 23 theme disease 18:24 arg1 mutations 26:34 Neurodegenerative disease mutations 0:34 Neurodegenerative disease mutations in TREM2 0:43 Neurodegenerative disease mutations in TREM2 reveal a functional surface and distinct loss-of-function mechanisms. 27995897 3 24 theme TREM2 588:592 arg1 function 594:601 TREM2 function 588:601 TREM2 function 588:601 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 1 25 theme neurodegenerative 193:209 arg1 diseases 211:218 several neurodegenerative diseases 185:218 several neurodegenerative diseases 185:218 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 0 26 theme Neurodegenerative 0:16 arg1 mutations 26:34 Neurodegenerative disease mutations 0:34 Neurodegenerative disease mutations in TREM2 0:43 Neurodegenerative disease mutations in TREM2 reveal a functional surface and distinct loss-of-function mechanisms. 27995897 4 27 theme risk 764:767 arg1 variants 769:776 Alzheimer's risk variants 752:776 Alzheimer's risk variants 752:776 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 2 28 theme wild-type 333:341 arg1 proteins 355:362 wild-type and variant proteins 333:362 wild-type and variant proteins 333:362 To determine how TREM2 variants contribute to these diseases, we performed structural and functional studies of wild-type and variant proteins. 27995897 4 29 theme folded 711:716 arg1 expression 732:741 folded TREM2 surface expression 711:741 folded TREM2 surface expression 711:741 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 6 30 theme structural 988:997 arg1 differences 1014:1024 structural and functional differences 988:1024 structural and functional differences among genetic variants of TREM2 988:1056 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 1 31 theme Genetic 115:121 arg1 variations 123:132 Genetic variations 115:132 Genetic variations in the myeloid immune receptor TREM2 115:169 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 3 32 theme risk 491:494 arg1 variants 496:503 Alzheimer's disease risk variants 471:503 Alzheimer's disease risk variants 471:503 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 6 33 theme medicine 1198:1205 arg1 approaches 1207:1216 patient-specific medicine approaches 1181:1216 patient-specific medicine approaches for neurodegenerative disorders 1181:1248 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 6 34 theme neurodegenerative 1222:1238 arg1 disorders 1240:1248 neurodegenerative disorders 1222:1248 neurodegenerative disorders 1222:1248 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 3 35 theme Nasu-Hakola 432:442 arg1 disease 444:450 Nasu-Hakola disease 432:450 Nasu-Hakola disease 432:450 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 6 36 theme functional 1153:1162 arg1 differences 1164:1174 these genetic and functional differences 1135:1174 these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders 1135:1248 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 6 37 theme TREM2 1099:1103 arg1 pathway 1105:1111 the TREM2 pathway 1095:1111 the TREM2 pathway 1095:1111 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 5 38 from surface 891:897 arg1 TREM2 902:906 TREM2 902:906 TREM2 902:906 Additionally, the Alzheimer's risk variants appear to epitope map a functional surface on TREM2 that is unique within the larger TREM family. 27995897 5 39 theme functional 880:889 arg1 unique 916:921 unique 916:921 unique 916:921 Additionally, the Alzheimer's risk variants appear to epitope map a functional surface on TREM2 that is unique within the larger TREM family. 27995897 5 39 theme functional 880:889 arg1 surface 891:897 a functional surface 878:897 a functional surface on TREM2 that is unique within the larger TREM family 878:951 Additionally, the Alzheimer's risk variants appear to epitope map a functional surface on TREM2 that is unique within the larger TREM family. 27995897 3 40 theme disease 483:489 arg1 variants 496:503 Alzheimer's disease risk variants 471:503 Alzheimer's disease risk variants 471:503 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 0 41 theme functional 54:63 arg1 surface 65:71 a functional surface 52:71 a functional surface 52:71 Neurodegenerative disease mutations in TREM2 reveal a functional surface and distinct loss-of-function mechanisms. 27995897 3 42 located found 423:427 arg2 mutations 413:421 mutations 413:421 mutations found in Nasu-Hakola disease 413:450 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 3 42 located found 423:427 arg1 disease 444:450 Nasu-Hakola disease 432:450 Nasu-Hakola disease 432:450 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 4 43 dep impact 673:678 arg1 whereas 744:750 whereas 744:750 whereas 744:750 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 5 44 theme larger 934:939 arg1 family 946:951 the larger TREM family 930:951 the larger TREM family 930:951 Additionally, the Alzheimer's risk variants appear to epitope map a functional surface on TREM2 that is unique within the larger TREM family. 27995897 2 45 theme TREM2 238:242 arg1 variants 244:251 TREM2 variants 238:251 TREM2 variants 238:251 To determine how TREM2 variants contribute to these diseases, we performed structural and functional studies of wild-type and variant proteins. 27995897 5 46 theme TREM 941:944 arg1 family 946:951 the larger TREM family 930:951 the larger TREM family 930:951 Additionally, the Alzheimer's risk variants appear to epitope map a functional surface on TREM2 that is unique within the larger TREM family. 27995897 3 47 contain have 563:566 arg1 mutations 553:561 these mutations 547:561 these mutations 547:561 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 3 47 contain have 563:566 arg2 effects 577:583 distinct effects 568:583 distinct effects 568:583 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 1 48 theme myeloid 141:147 arg1 TREM2 165:169 the myeloid immune receptor TREM2 137:169 the myeloid immune receptor TREM2 137:169 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 1 49 from variations 123:132 arg1 TREM2 165:169 the myeloid immune receptor TREM2 137:169 the myeloid immune receptor TREM2 137:169 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 1 50 theme immune 149:154 arg1 TREM2 165:169 the myeloid immune receptor TREM2 137:169 the myeloid immune receptor TREM2 137:169 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 2 51 theme proteins 355:362 arg1 studies 322:328 structural and functional studies 296:328 structural and functional studies of wild-type and variant proteins 296:362 To determine how TREM2 variants contribute to these diseases, we performed structural and functional studies of wild-type and variant proteins. 27995897 4 52 theme protein 680:686 arg1 stability 688:696 protein stability 680:696 protein stability 680:696 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 1 53 theme receptor 156:163 arg1 TREM2 165:169 the myeloid immune receptor TREM2 137:169 the myeloid immune receptor TREM2 137:169 Genetic variations in the myeloid immune receptor TREM2 are linked to several neurodegenerative diseases. 27995897 4 54 theme Nasu-Hakola 651:661 arg1 mutations 663:671 Nasu-Hakola mutations 651:671 Nasu-Hakola mutations 651:671 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 27995897 6 55 theme genetic 1141:1147 arg1 differences 1164:1174 these genetic and functional differences 1135:1174 these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders 1135:1248 These findings provide a guide to structural and functional differences among genetic variants of TREM2, indicating that therapies targeting the TREM2 pathway should be tailored to these genetic and functional differences with patient-specific medicine approaches for neurodegenerative disorders. 27995897 3 56 located found 509:513 arg2 variants 496:503 Alzheimer's disease risk variants 471:503 Alzheimer's disease risk variants 471:503 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 3 56 located found 509:513 arg1 surface 522:528 the surface 518:528 the surface 518:528 Our 3.1 Å TREM2 crystal structure revealed that mutations found in Nasu-Hakola disease are buried whereas Alzheimer's disease risk variants are found on the surface, suggesting that these mutations have distinct effects on TREM2 function. 27995897 4 57 theme surface 724:730 arg1 expression 732:741 folded TREM2 surface expression 711:741 folded TREM2 surface expression 711:741 Biophysical and cellular methods indicate that Nasu-Hakola mutations impact protein stability and decrease folded TREM2 surface expression, whereas Alzheimer's risk variants impact binding to a TREM2 ligand. 19358553 4 0 theme N-glycans 931:939 arg1 forms 922:926 various forms 914:926 various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5) 914:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 7 1 theme structure/function 1891:1908 arg1 relationships 1910:1922 glycan structure/function relationships 1884:1922 glycan structure/function relationships 1884:1922 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 4 2 theme peak 1121:1124 arg1 intensities 1126:1136 peak intensities 1121:1136 peak intensities 1121:1136 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 6 3 theme instrumental 1725:1736 arg1 settings 1738:1745 instrumental settings 1725:1745 instrumental settings 1725:1745 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 6 4 theme MS 1505:1506 arg1 strength 1515:1522 MALDI-TOF MS signal strength 1495:1522 MALDI-TOF MS signal strength of glycopeptides 1495:1539 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 3 5 dep quantitation 744:755 arg1 = 796:796 = 796:796 = 796:796 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 6 theme neutral 628:634 arg1 glycans 636:642 neutral glycans 628:642 neutral glycans (ribonuclease B, IgG, and ovalbumin) 628:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 6 theme neutral 628:634 arg1 B 658:658 ribonuclease B 645:658 ribonuclease B 645:658 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 6 theme neutral 628:634 arg1 ovalbumin 670:678 ovalbumin 670:678 ovalbumin 670:678 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 6 theme neutral 628:634 arg1 IgG 661:663 IgG 661:663 IgG 661:663 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 6 7 theme relative 1582:1589 arg1 quantities 1591:1600 the relative quantities 1578:1600 the relative quantities of glycoforms 1578:1614 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 1 8 theme laser 221:225 arg1 spectrometry 269:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 205:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 205:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 8 theme laser 221:225 arg1 MS 293:294 MALDI-TOF MS 283:294 MALDI-TOF MS 283:294 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 7 9 theme biomarker 1845:1853 arg1 discovery 1855:1863 biomarker discovery 1845:1863 biomarker discovery 1845:1863 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 4 10 theme acids 1205:1209 arg1 loss 1190:1193 only a minor loss 1177:1193 only a minor loss of sialic acids (r = 0.9664, n = 5) 1177:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 5 11 theme critical 1460:1467 arg1 parameter 1469:1477 a critical parameter 1458:1477 a critical parameter 1458:1477 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 5 11 theme critical 1460:1467 arg1 capacity 1446:1453 the column capacity 1435:1453 the column capacity 1435:1453 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 1 12 theme time-of-flight 249:262 arg1 spectrometry 269:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 205:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 205:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 12 theme time-of-flight 249:262 arg1 MS 293:294 MALDI-TOF MS 283:294 MALDI-TOF MS 283:294 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 4 13 theme minor 1184:1188 arg1 loss 1190:1193 only a minor loss 1177:1193 only a minor loss of sialic acids (r = 0.9664, n = 5) 1177:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 3 14 theme HPLC 839:842 arg1 profile 856:862 a normal phase HPLC 2-AB glycan profile 824:862 a normal phase HPLC 2-AB glycan profile 824:862 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 1 15 theme MALDI-TOF 283:291 arg1 spectrometry 269:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 205:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 205:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 15 theme MALDI-TOF 283:291 arg1 MS 293:294 MALDI-TOF MS 283:294 MALDI-TOF MS 283:294 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 3 16 dep = 784:784 arg1 0.9958 786:791 0.9958 786:791 0.9958 786:791 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 17 theme normal 826:831 arg1 profile 856:862 a normal phase HPLC 2-AB glycan profile 824:862 a normal phase HPLC 2-AB glycan profile 824:862 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 6 18 theme technical 1640:1648 arg1 issues 1650:1655 certain technical issues 1632:1655 certain technical issues 1632:1655 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 1 19 gly glycopeptides 185:197 arg2 glycopeptides 185:197 N-linked glycopeptides 176:197 N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 176:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 7 20 theme site-specific 1781:1793 arg1 glycoprofiling 1795:1808 rapid and sensitive site-specific glycoprofiling 1761:1808 rapid and sensitive site-specific glycoprofiling of N-glycan populations 1761:1832 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 1 21 theme Site-specific 144:156 arg1 technique 312:320 an emerging technique 300:320 an emerging technique 300:320 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 21 theme Site-specific 144:156 arg1 glycoprofiling 158:171 Site-specific glycoprofiling 144:171 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 144:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 6 22 theme matrix 1706:1711 arg1 choice 1713:1718 matrix choice 1706:1718 matrix choice 1706:1718 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 3 23 theme glycan 849:854 arg1 profile 856:862 a normal phase HPLC 2-AB glycan profile 824:862 a normal phase HPLC 2-AB glycan profile 824:862 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 1 24 theme N-linked 176:183 arg1 glycopeptides 185:197 N-linked glycopeptides 176:197 N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 176:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 2 25 theme signal 559:564 arg1 strength 566:573 the MALDI-TOF MS signal strength 542:573 the MALDI-TOF MS signal strength 542:573 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 3 26 gly glycoproteins 603:615 arg1 glycoproteins 603:615 glycoproteins 603:615 glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) 603:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 5 27 theme materials 1299:1307 arg1 range 1265:1269 a range 1263:1269 a range of hydrophilic and graphite materials packed in microcolumn format 1263:1336 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 0 28 theme signal 102:107 arg1 strength 109:116 signal strength 102:116 signal strength 102:116 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 6 29 theme sample 1689:1694 arg1 handling 1696:1703 nonbiased sample handling 1679:1703 nonbiased sample handling 1679:1703 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 4 30 gly sialoglycopeptides 1028:1045 arg2 sialoglycopeptides 1028:1045 three different sialoglycopeptides 1012:1045 three different sialoglycopeptides from fetuin 1012:1057 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 4 30 gly sialoglycopeptides 1028:1045 arg1 fetuin 1052:1057 fetuin 1052:1057 fetuin 1052:1057 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 2 31 theme individual 516:525 arg1 glycoforms 527:536 the individual glycoforms 512:536 the individual glycoforms 512:536 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 0 32 theme Site-specific 0:12 arg1 glycoprofiling 14:27 Site-specific glycoprofiling 0:27 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 0:142 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 2 33 gly glycopeptides 419:431 arg2 glycopeptides 419:431 widely different glycopeptides 402:431 widely different glycopeptides 402:431 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 5 34 gly glycopeptide 1236:1247 arg2 glycopeptide 1236:1247 glycopeptide purification 1236:1260 glycopeptide purification 1236:1260 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 2 35 theme relative 489:496 arg1 abundances 498:507 the relative abundances 485:507 the relative abundances of the individual glycoforms 485:536 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 2 36 theme MALDI-TOF 546:554 arg1 MS 556:557 MALDI-TOF MS 546:557 the MALDI-TOF MS signal strength 542:573 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 3 37 theme reproducible 731:742 arg1 quantitation 744:755 excellent and reproducible quantitation 717:755 excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) 717:799 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 4 38 theme precise 876:882 arg1 quantitation 884:895 precise quantitation 876:895 precise quantitation 876:895 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 0 39 theme glycopeptides 41:53 arg1 glycoprofiling 14:27 Site-specific glycoprofiling 0:27 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 0:142 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 3 40 contain containing 617:626 arg2 IgG 661:663 IgG 661:663 IgG 661:663 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 40 contain containing 617:626 arg2 B 658:658 ribonuclease B 645:658 ribonuclease B 645:658 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 40 contain containing 617:626 arg2 glycans 636:642 neutral glycans 628:642 neutral glycans (ribonuclease B, IgG, and ovalbumin) 628:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 40 contain containing 617:626 arg2 ovalbumin 670:678 ovalbumin 670:678 ovalbumin 670:678 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 40 contain containing 617:626 arg1 glycoproteins 603:615 glycoproteins 603:615 glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) 603:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 41 theme excellent 717:725 arg1 quantitation 744:755 excellent and reproducible quantitation 717:755 excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) 717:799 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 0 42 theme MALDI-TOF 61:69 arg1 MS 71:72 MALDI-TOF MS 61:72 MALDI-TOF MS 61:72 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 3 43 attach derived 590:596 arg2 Glycopeptides 576:588 Glycopeptides 576:588 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) 576:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 43 attach derived 590:596 arg1 glycoproteins 603:615 glycoproteins 603:615 glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) 603:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 44 theme coefficient 770:780 arg1 r 782:782 correlation coefficient r 758:782 correlation coefficient r = 0.9958 758:791 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 2 45 theme systematic 382:391 arg1 study 393:397 a systematic study 380:397 a systematic study of widely different glycopeptides 380:431 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 4 46 from fetuin 1052:1057 arg1 sialoglycopeptides 1028:1045 three different sialoglycopeptides 1012:1045 three different sialoglycopeptides from fetuin 1012:1057 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 7 47 theme relationships 1910:1922 arg1 elucidation 1869:1879 elucidation 1869:1879 elucidation of glycan structure/function relationships 1869:1922 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 7 47 theme relationships 1910:1922 arg1 discovery 1855:1863 biomarker discovery 1845:1863 biomarker discovery 1845:1863 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 5 48 theme information 1406:1416 arg1 loss 1385:1388 loss 1385:1388 loss of quantitative information 1385:1416 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 7 49 theme glycan 1884:1889 arg1 relationships 1910:1922 glycan structure/function relationships 1884:1922 glycan structure/function relationships 1884:1922 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 2 50 theme different 409:417 arg1 glycopeptides 419:431 widely different glycopeptides 402:431 widely different glycopeptides 402:431 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 6 51 theme signal 1508:1513 arg1 strength 1515:1522 MALDI-TOF MS signal strength 1495:1522 MALDI-TOF MS signal strength of glycopeptides 1495:1539 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 1 52 theme emerging 303:310 arg1 technique 312:320 an emerging technique 300:320 an emerging technique 300:320 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 52 theme emerging 303:310 arg1 glycoprofiling 158:171 Site-specific glycoprofiling 144:171 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 144:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 0 53 link N-linked 32:39 arg1 glycopeptides 41:53 N-linked glycopeptides 32:53 N-linked glycopeptides using MALDI-TOF MS 32:72 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 4 54 theme relative 1142:1149 arg1 abundances 1151:1160 relative abundances 1142:1160 relative abundances 1142:1160 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 3 55 theme = 784:784 arg1 r 782:782 correlation coefficient r 758:782 correlation coefficient r = 0.9958 758:791 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 6 56 theme MALDI-TOF 1495:1503 arg1 MS 1505:1506 MALDI-TOF MS 1495:1506 MALDI-TOF MS signal strength of glycopeptides 1495:1539 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 5 57 theme glycopeptide 1236:1247 arg1 purification 1249:1260 glycopeptide purification 1236:1260 glycopeptide purification 1236:1260 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 4 58 theme sialic 1198:1203 arg1 r 1212:1212 r = 0.9664 1212:1221 r = 0.9664 1212:1221 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 4 58 theme sialic 1198:1203 arg1 acids 1205:1209 sialic acids 1198:1209 sialic acids (r = 0.9664, n = 5) 1198:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 1 59 theme matrix-assisted 205:219 arg1 spectrometry 269:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 205:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 205:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 59 theme matrix-assisted 205:219 arg1 MS 293:294 MALDI-TOF MS 283:294 MALDI-TOF MS 283:294 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 4 60 dep r 1212:1212 arg1 n 1224:1224 n = 5 1224:1228 n = 5 1224:1228 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 1 61 theme desorption/ionization 227:247 arg1 spectrometry 269:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 205:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 205:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 61 theme desorption/ionization 227:247 arg1 MS 293:294 MALDI-TOF MS 283:294 MALDI-TOF MS 283:294 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 4 62 theme various 914:920 arg1 forms 922:926 various forms 914:926 various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5) 914:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 7 63 theme populations 1822:1832 arg1 glycoprofiling 1795:1808 rapid and sensitive site-specific glycoprofiling 1761:1808 rapid and sensitive site-specific glycoprofiling of N-glycan populations 1761:1832 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 3 64 theme ribonuclease 645:656 arg1 glycans 636:642 neutral glycans 628:642 neutral glycans (ribonuclease B, IgG, and ovalbumin) 628:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 64 theme ribonuclease 645:656 arg1 B 658:658 ribonuclease B 645:658 ribonuclease B 645:658 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 1 65 theme mass 264:267 arg1 spectrometry 269:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 205:280 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 205:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 65 theme mass 264:267 arg1 MS 293:294 MALDI-TOF MS 283:294 MALDI-TOF MS 283:294 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 4 66 theme = 1226:1226 arg1 n 1224:1224 n = 5 1224:1228 n = 5 1224:1228 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 3 67 theme 2-AB 844:847 arg1 profile 856:862 a normal phase HPLC 2-AB glycan profile 824:862 a normal phase HPLC 2-AB glycan profile 824:862 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 68 theme phase 833:837 arg1 profile 856:862 a normal phase HPLC 2-AB glycan profile 824:862 a normal phase HPLC 2-AB glycan profile 824:862 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 6 69 theme certain 1632:1638 arg1 issues 1650:1655 certain technical issues 1632:1655 certain technical issues 1632:1655 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 3 70 dep glycans 636:642 arg1 ovalbumin 670:678 ovalbumin 670:678 ovalbumin 670:678 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 70 dep glycans 636:642 arg1 B 658:658 ribonuclease B 645:658 ribonuclease B 645:658 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 70 dep glycans 636:642 arg1 glycans 636:642 neutral glycans 628:642 neutral glycans (ribonuclease B, IgG, and ovalbumin) 628:679 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 3 70 dep glycans 636:642 arg1 IgG 661:663 IgG 661:663 IgG 661:663 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 7 71 theme N-glycan 1813:1820 arg1 populations 1822:1832 N-glycan populations 1813:1832 N-glycan populations 1813:1832 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 4 72 theme = 1214:1214 arg1 r 1212:1212 r = 0.9664 1212:1221 r = 0.9664 1212:1221 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 4 72 theme = 1214:1214 arg1 acids 1205:1209 sialic acids 1198:1209 sialic acids (r = 0.9664, n = 5) 1198:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 5 73 theme microcolumn 1319:1329 arg1 format 1331:1336 microcolumn format 1319:1336 microcolumn format 1319:1336 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 0 74 theme strong 75:80 arg1 correlation 82:92 strong correlation 75:92 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 0:142 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 4 75 theme different 1018:1026 arg1 sialoglycopeptides 1028:1045 three different sialoglycopeptides 1012:1045 three different sialoglycopeptides from fetuin 1012:1057 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 5 76 theme hydrophilic 1274:1284 arg1 materials 1299:1307 hydrophilic and graphite materials 1274:1307 hydrophilic and graphite materials packed in microcolumn format 1274:1336 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 6 77 theme glycoforms 1605:1614 arg1 quantities 1591:1600 the relative quantities 1578:1600 the relative quantities of glycoforms 1578:1614 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 1 78 theme glycopeptides 185:197 arg1 technique 312:320 an emerging technique 300:320 an emerging technique 300:320 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 1 78 theme glycopeptides 185:197 arg1 glycoprofiling 158:171 Site-specific glycoprofiling 144:171 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 144:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 7 79 theme sensitive 1771:1779 arg1 glycoprofiling 1795:1808 rapid and sensitive site-specific glycoprofiling 1761:1808 rapid and sensitive site-specific glycoprofiling of N-glycan populations 1761:1832 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 2 80 theme MS 556:557 arg1 strength 566:573 the MALDI-TOF MS signal strength 542:573 the MALDI-TOF MS signal strength 542:573 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 5 81 theme graphite 1290:1297 arg1 materials 1299:1307 hydrophilic and graphite materials 1274:1307 hydrophilic and graphite materials packed in microcolumn format 1274:1336 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 4 82 dep MS. 995:997 arg1 profiled 1082:1089 profiled 1082:1089 were site-specifically profiled 1059:1089 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 4 82 dep MS. 995:997 arg1 found 1166:1170 found 1166:1170 was found with only a minor loss of sialic acids (r = 0.9664, n = 5) 1162:1229 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 6 83 located found 1550:1554 arg2 strength 1515:1522 MALDI-TOF MS signal strength 1495:1522 MALDI-TOF MS signal strength of glycopeptides 1495:1539 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 6 83 located found 1550:1554 arg1 conclusion 1483:1492 conclusion 1483:1492 conclusion 1483:1492 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 7 84 theme rapid 1761:1765 arg1 glycoprofiling 1795:1808 rapid and sensitive site-specific glycoprofiling 1761:1808 rapid and sensitive site-specific glycoprofiling of N-glycan populations 1761:1832 This enables rapid and sensitive site-specific glycoprofiling of N-glycan populations to promote biomarker discovery and elucidation of glycan structure/function relationships. 19358553 0 85 theme glycoform 122:130 arg1 quantities 132:141 glycoform quantities 122:141 glycoform quantities 122:141 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 6 86 theme nonbiased 1679:1687 arg1 handling 1696:1703 nonbiased sample handling 1679:1703 nonbiased sample handling 1679:1703 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 6 87 gly glycopeptides 1527:1539 arg2 glycopeptides 1527:1539 glycopeptides 1527:1539 glycopeptides 1527:1539 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 0 88 theme N-linked 32:39 arg1 glycopeptides 41:53 N-linked glycopeptides 32:53 N-linked glycopeptides using MALDI-TOF MS 32:72 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 1 89 theme quantitative 331:342 arg1 accuracy 344:351 its quantitative accuracy 327:351 its quantitative accuracy 327:351 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 6 90 dep handling 1696:1703 arg1 i.e. 1673:1676 i.e. 1673:1676 i.e. 1673:1676 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 1 91 link N-linked 176:183 arg1 glycopeptides 185:197 N-linked glycopeptides 176:197 N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 176:295 Site-specific glycoprofiling of N-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging technique, but its quantitative accuracy lacks documentation. 19358553 4 92 dep N-glycans 931:939 arg1 permethylated 948:960 permethylated 948:960 permethylated 948:960 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 4 92 dep N-glycans 931:939 arg1 fluorescence-labeled 967:986 fluorescence-labeled 967:986 fluorescence-labeled 967:986 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 4 92 dep N-glycans 931:939 arg1 free 942:945 free 942:945 free 942:945 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 19358553 3 93 dep = 796:796 arg1 r 782:782 correlation coefficient r 758:782 correlation coefficient r = 0.9958 758:791 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 2 94 theme glycoforms 527:536 arg1 strength 566:573 the MALDI-TOF MS signal strength 542:573 the MALDI-TOF MS signal strength 542:573 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 2 94 theme glycoforms 527:536 arg1 abundances 498:507 the relative abundances 485:507 the relative abundances of the individual glycoforms 485:536 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 3 95 theme correlation 758:768 arg1 r 782:782 correlation coefficient r 758:782 correlation coefficient r = 0.9958 758:791 Glycopeptides derived from glycoproteins containing neutral glycans (ribonuclease B, IgG, and ovalbumin) were initially profiled and yielded excellent and reproducible quantitation (correlation coefficient r = 0.9958, n = 5) when evaluated against a normal phase HPLC 2-AB glycan profile. 19358553 5 96 theme column 1439:1444 arg1 parameter 1469:1477 a critical parameter 1458:1477 a critical parameter 1458:1477 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 5 96 theme column 1439:1444 arg1 capacity 1446:1453 the column capacity 1435:1453 the column capacity 1435:1453 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 0 97 gly glycopeptides 41:53 arg2 glycopeptides 41:53 N-linked glycopeptides 32:53 N-linked glycopeptides using MALDI-TOF MS 32:72 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 5 98 theme quantitative 1393:1404 arg1 information 1406:1416 quantitative information 1393:1416 quantitative information 1393:1416 For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format proved capable of performing desalting without loss of quantitative information, but highlighted the column capacity as a critical parameter. 19358553 2 99 theme glycopeptides 419:431 arg1 study 393:397 a systematic study 380:397 a systematic study of widely different glycopeptides 380:431 Thus, a systematic study of widely different glycopeptides was performed to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. 19358553 6 100 theme glycopeptides 1527:1539 arg1 strength 1515:1522 MALDI-TOF MS signal strength 1495:1522 MALDI-TOF MS signal strength of glycopeptides 1495:1539 In conclusion, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, providing that certain technical issues are considered, i.e., nonbiased sample handling, matrix choice, and instrumental settings. 19358553 0 101 dep glycoprofiling 14:27 arg1 correlation 82:92 strong correlation 75:92 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 0:142 Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: strong correlation between signal strength and glycoform quantities. 19358553 4 102 theme good 1096:1099 arg1 correlation 1101:1111 good correlation 1096:1111 good correlation between peak intensities and relative abundances 1096:1160 Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialoglycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r = 0.9664, n = 5). 21156796 0 0 theme receptor 79:86 arg1 triggering 88:97 cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering 37:97 cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering 37:97 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 4 1 theme 1.8-Å 694:698 arg1 structure 711:719 The 1.8-Å resolution structure 690:719 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer 690:752 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 1 2 theme lymphocyte-associated 162:182 arg1 receptor 140:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 2 theme lymphocyte-associated 162:182 arg1 CTLA-4 195:200 CTLA-4 195:200 CTLA-4 195:200 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 2 theme lymphocyte-associated 162:182 arg1 antigen-4 184:192 cytotoxic T lymphocyte-associated antigen-4 150:192 cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) 150:201 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 3 theme proteins 247:254 arg1 class 225:229 the class 221:229 the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors 221:336 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 8 4 theme CTLA-4 1536:1541 arg1 phosphorylation 1543:1557 CTLA-4 phosphorylation 1536:1557 CTLA-4 phosphorylation by extrinsic kinase(s) 1536:1580 The new structure establishes that rigid-body ligand interactions are capable of triggering CTLA-4 phosphorylation by extrinsic kinase(s). 21156796 1 5 theme related 349:355 arg1 B7-1 366:369 B7-1 366:369 B7-1 366:369 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 5 theme related 349:355 arg1 ligands 357:363 the related ligands 345:363 the related ligands 345:363 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 5 theme related 349:355 arg1 B7-2 375:378 B7-2 375:378 B7-2 375:378 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 8 6 theme extrinsic 1562:1570 arg1 s 1579:1579 extrinsic kinase(s) 1562:1580 extrinsic kinase(s) 1562:1580 The new structure establishes that rigid-body ligand interactions are capable of triggering CTLA-4 phosphorylation by extrinsic kinase(s). 21156796 4 7 theme resolution 700:709 arg1 structure 711:719 The 1.8-Å resolution structure 690:719 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer 690:752 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 5 8 from contrast 979:986 arg1 similar 960:966 similar 960:966 similar 960:966 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 5 8 from contrast 979:986 arg1 forms 915:919 The ligand-bound and unbound forms 886:919 The ligand-bound and unbound forms of both CTLA-4 and B7-1 886:943 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 8 9 theme rigid-body 1479:1488 arg1 interactions 1497:1508 rigid-body ligand interactions 1479:1508 rigid-body ligand interactions 1479:1508 The new structure establishes that rigid-body ligand interactions are capable of triggering CTLA-4 phosphorylation by extrinsic kinase(s). 21156796 7 10 theme parameters 1230:1239 arg1 similarity 1198:1207 The similarity 1194:1207 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 1194:1300 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 3 11 theme ligand-bound 553:564 arg1 CTLA-4 566:571 ligand-bound CTLA-4 553:571 ligand-bound CTLA-4 553:571 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 5 12 theme marked 972:977 arg1 contrast 979:986 marked contrast 972:986 marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit 972:1047 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 7 13 theme selectivity 1431:1441 arg1 level 1422:1426 some level 1417:1426 some level of selectivity 1417:1441 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 4 14 theme shared 769:774 arg1 history 789:795 the shared evolutionary history 765:795 the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors 765:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 3 15 theme CTLA-4 566:571 arg1 structures 539:548 Crystal structures 531:548 Crystal structures of ligand-bound CTLA-4 531:571 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 1 16 theme extrinsic 274:282 arg1 kinases 293:299 extrinsic tyrosine kinases 274:299 extrinsic tyrosine kinases that also includes antigen receptors 274:336 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 2 17 theme receptors 520:528 arg1 class 511:515 this class 506:515 this class of receptors 506:528 Conformational changes are commonly invoked to explain ligand-induced "triggering" of this class of receptors. 21156796 8 18 theme kinase 1572:1577 arg1 s 1579:1579 extrinsic kinase(s) 1562:1580 extrinsic kinase(s) 1562:1580 The new structure establishes that rigid-body ligand interactions are capable of triggering CTLA-4 phosphorylation by extrinsic kinase(s). 21156796 4 19 theme antigen 867:873 arg1 receptors 875:883 the antigen receptors 863:883 the antigen receptors 863:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 6 20 theme Isothermal 1050:1059 arg1 calorimetry 1071:1081 Isothermal titration calorimetry 1050:1081 Isothermal titration calorimetry 1050:1081 Isothermal titration calorimetry reveals that ligand binding by CTLA-4 is enthalpically driven and accompanied by unfavorable entropic changes. 21156796 6 21 theme ligand 1096:1101 arg1 binding 1103:1109 ligand binding 1096:1109 ligand binding by CTLA-4 1096:1119 Isothermal titration calorimetry reveals that ligand binding by CTLA-4 is enthalpically driven and accompanied by unfavorable entropic changes. 21156796 7 22 theme thermodynamic 1216:1228 arg1 parameters 1230:1239 the thermodynamic parameters 1212:1239 the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 1212:1300 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 5 23 theme CTLA-4 929:934 arg1 forms 915:919 The ligand-bound and unbound forms 886:919 The ligand-bound and unbound forms of both CTLA-4 and B7-1 886:943 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 5 23 theme CTLA-4 929:934 arg1 similar 960:966 similar 960:966 similar 960:966 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 2 24 theme triggering 491:500 arg1 " 501:501 ligand-induced "triggering" 475:501 ligand-induced "triggering" of this class of receptors 475:528 Conformational changes are commonly invoked to explain ligand-induced "triggering" of this class of receptors. 21156796 5 25 theme ligand-bound 890:901 arg1 forms 915:919 The ligand-bound and unbound forms 886:919 The ligand-bound and unbound forms of both CTLA-4 and B7-1 886:943 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 5 25 theme ligand-bound 890:901 arg1 similar 960:966 similar 960:966 similar 960:966 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 0 26 theme ligand 11:16 arg1 recognition 18:28 Rigid-body ligand recognition 0:28 Rigid-body ligand recognition 0:28 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 4 27 theme homodimer 744:752 arg1 structure 711:719 The 1.8-Å resolution structure 690:719 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer 690:752 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 0 28 theme Rigid-body 0:9 arg1 recognition 18:28 Rigid-body ligand recognition 0:28 Rigid-body ligand recognition 0:28 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 1 29 theme tyrosine 284:291 arg1 kinases 293:299 extrinsic tyrosine kinases 274:299 extrinsic tyrosine kinases that also includes antigen receptors 274:336 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 4 30 theme apo 727:729 arg1 homodimer 744:752 an apo human CTLA-4 homodimer 724:752 an apo human CTLA-4 homodimer 724:752 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 4 31 theme CTLA-4 737:742 arg1 homodimer 744:752 an apo human CTLA-4 homodimer 724:752 an apo human CTLA-4 homodimer 724:752 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 5 32 theme unbound 907:913 arg1 forms 915:919 The ligand-bound and unbound forms 886:919 The ligand-bound and unbound forms of both CTLA-4 and B7-1 886:943 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 5 32 theme unbound 907:913 arg1 similar 960:966 similar 960:966 similar 960:966 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 7 33 with interactions 1260:1271 arg1 B7-1 1288:1291 B7-1 1288:1291 B7-1 1288:1291 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 7 33 with interactions 1260:1271 arg1 B7-2 1297:1300 B7-2 1297:1300 B7-2 1297:1300 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 8 34 theme ligand 1490:1495 arg1 interactions 1497:1508 rigid-body ligand interactions 1479:1508 rigid-body ligand interactions 1479:1508 The new structure establishes that rigid-body ligand interactions are capable of triggering CTLA-4 phosphorylation by extrinsic kinase(s). 21156796 5 35 theme induced 1037:1043 arg1 fit 1045:1047 induced fit 1037:1047 induced fit 1037:1047 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 6 36 theme unfavorable 1164:1174 arg1 changes 1185:1191 unfavorable entropic changes 1164:1191 unfavorable entropic changes 1164:1191 Isothermal titration calorimetry reveals that ligand binding by CTLA-4 is enthalpically driven and accompanied by unfavorable entropic changes. 21156796 6 37 theme titration 1061:1069 arg1 calorimetry 1071:1081 Isothermal titration calorimetry 1050:1081 Isothermal titration calorimetry 1050:1081 Isothermal titration calorimetry reveals that ligand binding by CTLA-4 is enthalpically driven and accompanied by unfavorable entropic changes. 21156796 6 38 theme entropic 1176:1183 arg1 changes 1185:1191 unfavorable entropic changes 1164:1191 unfavorable entropic changes 1164:1191 Isothermal titration calorimetry reveals that ligand binding by CTLA-4 is enthalpically driven and accompanied by unfavorable entropic changes. 21156796 3 39 theme structural 642:651 arg1 changes 653:659 the structural changes 638:659 the structural changes accompanying ligand binding 638:687 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 1 40 theme inhibitory 104:113 arg1 receptor 140:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 40 theme inhibitory 104:113 arg1 antigen-4 184:192 cytotoxic T lymphocyte-associated antigen-4 150:192 cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) 150:201 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 2 41 theme Conformational 420:433 arg1 changes 435:441 Conformational changes 420:441 Conformational changes 420:441 Conformational changes are commonly invoked to explain ligand-induced "triggering" of this class of receptors. 21156796 0 42 theme T-lymphocyte 47:58 arg1 antigen 60:66 cytotoxic T-lymphocyte antigen 4 37:68 cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering 37:97 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 0 42 theme T-lymphocyte 47:58 arg1 CTLA-4 71:76 CTLA-4 71:76 CTLA-4 71:76 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 5 43 theme B7-1 940:943 arg1 forms 915:919 The ligand-bound and unbound forms 886:919 The ligand-bound and unbound forms of both CTLA-4 and B7-1 886:943 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 5 43 theme B7-1 940:943 arg1 similar 960:966 similar 960:966 similar 960:966 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 3 44 theme changes 653:659 arg1 analysis 626:633 analysis 626:633 analysis of the structural changes accompanying ligand binding 626:687 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 4 45 theme immunoglobulin 832:845 arg1 superfamily 847:857 the immunoglobulin superfamily 828:857 the immunoglobulin superfamily 828:857 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 0 46 theme cytotoxic 37:45 arg1 antigen 60:66 cytotoxic T-lymphocyte antigen 4 37:68 cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering 37:97 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 0 46 theme cytotoxic 37:45 arg1 CTLA-4 71:76 CTLA-4 71:76 CTLA-4 71:76 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 3 47 theme apo 605:607 arg1 form 609:612 the apo form 601:612 the apo form 601:612 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 2 48 theme ligand-induced 475:488 arg1 " 501:501 ligand-induced "triggering" 475:501 ligand-induced "triggering" of this class of receptors 475:528 Conformational changes are commonly invoked to explain ligand-induced "triggering" of this class of receptors. 21156796 1 49 theme antigen 320:326 arg1 receptors 328:336 antigen receptors 320:336 antigen receptors 320:336 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 50 theme antigen-presenting 394:411 arg1 cells 413:417 antigen-presenting cells 394:417 antigen-presenting cells 394:417 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 7 51 theme B7-2 1396:1399 arg1 binding 1401:1407 B7-2 binding 1396:1407 B7-2 binding 1396:1407 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 4 52 theme receptors 875:883 arg1 subgroup 816:823 the CTLA-4/CD28 subgroup 800:823 the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors 800:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 1 53 theme T-cell 115:120 arg1 receptor 140:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 53 theme T-cell 115:120 arg1 antigen-4 184:192 cytotoxic T lymphocyte-associated antigen-4 150:192 cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) 150:201 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 0 54 theme antigen 60:66 arg1 triggering 88:97 cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering 37:97 cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering 37:97 Rigid-body ligand recognition drives cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor triggering. 21156796 5 55 theme fit 1045:1047 arg1 elements 1025:1032 elements 1025:1032 elements of induced fit 1025:1047 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 4 56 theme superfamily 847:857 arg1 subgroup 816:823 the CTLA-4/CD28 subgroup 800:823 the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors 800:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 1 57 theme surface-expressed 122:138 arg1 receptor 140:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 57 theme surface-expressed 122:138 arg1 antigen-4 184:192 cytotoxic T lymphocyte-associated antigen-4 150:192 cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) 150:201 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 8 58 theme new 1448:1450 arg1 structure 1452:1460 The new structure 1444:1460 The new structure 1444:1460 The new structure establishes that rigid-body ligand interactions are capable of triggering CTLA-4 phosphorylation by extrinsic kinase(s). 21156796 4 59 theme CTLA-4/CD28 804:814 arg1 subgroup 816:823 the CTLA-4/CD28 subgroup 800:823 the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors 800:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 4 60 theme human 731:735 arg1 homodimer 744:752 an apo human CTLA-4 homodimer 724:752 an apo human CTLA-4 homodimer 724:752 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 4 61 theme evolutionary 776:787 arg1 history 789:795 the shared evolutionary history 765:795 the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors 765:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 3 62 theme Crystal 531:537 arg1 structures 539:548 Crystal structures 531:548 Crystal structures of ligand-bound CTLA-4 531:571 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 5 63 from similar 960:966 arg1 contrast 979:986 marked contrast 972:986 marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit 972:1047 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 2 64 theme class 511:515 arg1 " 501:501 ligand-induced "triggering" 475:501 ligand-induced "triggering" of this class of receptors 475:528 Conformational changes are commonly invoked to explain ligand-induced "triggering" of this class of receptors. 21156796 3 65 theme ligand 674:679 arg1 binding 681:687 ligand binding 674:687 ligand binding 674:687 Crystal structures of ligand-bound CTLA-4 have been reported, but not the apo form, precluding analysis of the structural changes accompanying ligand binding. 21156796 7 66 theme conformational 1360:1373 arg1 changes 1375:1381 the conformational changes 1356:1381 the conformational changes observed for B7-2 binding 1356:1407 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 1 67 theme cytotoxic 150:158 arg1 receptor 140:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 67 theme cytotoxic 150:158 arg1 CTLA-4 195:200 CTLA-4 195:200 CTLA-4 195:200 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 67 theme cytotoxic 150:158 arg1 antigen-4 184:192 cytotoxic T lymphocyte-associated antigen-4 150:192 cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) 150:201 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 5 68 contain has 1021:1023 arg2 elements 1025:1032 elements 1025:1032 elements of induced fit 1025:1047 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 5 68 contain has 1021:1023 arg1 binding 1003:1009 binding 1003:1009 binding to CTLA-4 1003:1019 The ligand-bound and unbound forms of both CTLA-4 and B7-1 are remarkably similar, in marked contrast to B7-2, whose binding to CTLA-4 has elements of induced fit. 21156796 1 69 theme cell 234:237 arg1 proteins 247:254 cell surface proteins 234:254 cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors 234:336 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 4 70 theme subgroup 816:823 arg1 history 789:795 the shared evolutionary history 765:795 the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors 765:883 The 1.8-Å resolution structure of an apo human CTLA-4 homodimer emphasizes the shared evolutionary history of the CTLA-4/CD28 subgroup of the immunoglobulin superfamily and the antigen receptors. 21156796 1 71 theme T 160:160 arg1 receptor 140:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor 100:147 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 71 theme T 160:160 arg1 CTLA-4 195:200 CTLA-4 195:200 CTLA-4 195:200 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 1 71 theme T 160:160 arg1 antigen-4 184:192 cytotoxic T lymphocyte-associated antigen-4 150:192 cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) 150:201 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 21156796 7 72 theme CTLA-4 1276:1281 arg1 interactions 1260:1271 the interactions 1256:1271 the interactions of CTLA-4 with B7-1 and B7-2 1256:1300 The similarity of the thermodynamic parameters determined for the interactions of CTLA-4 with B7-1 and B7-2 suggests that the binding is not highly specific, but the conformational changes observed for B7-2 binding suggest some level of selectivity. 21156796 1 73 theme surface 239:245 arg1 proteins 247:254 cell surface proteins 234:254 cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors 234:336 The inhibitory T-cell surface-expressed receptor, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which belongs to the class of cell surface proteins phosphorylated by extrinsic tyrosine kinases that also includes antigen receptors, binds the related ligands, B7-1 and B7-2, expressed on antigen-presenting cells. 27316455 0 0 theme endoplasmic 72:82 arg1 reticulum 84:92 the endoplasmic reticulum 68:92 the endoplasmic reticulum 68:92 Molecular determinants that mediate the sorting of human ATG9A from the endoplasmic reticulum. 27316455 3 1 theme present 323:329 arg1 work 331:334 the present work 319:334 the present work 319:334 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 7 2 theme LYM/AAA 1042:1048 arg1 mutation 1050:1057 the LYM/AAA mutation 1038:1057 the LYM/AAA mutation 1038:1057 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 1 3 theme autophagosome 150:162 arg1 formation 164:172 autophagosome formation 150:172 autophagosome formation 150:172 ATG9A is a multispanning membrane protein required for autophagosome formation. 27316455 6 4 with oligomerization 828:842 arg1 ATG9A 896:900 co-transfected wild-type ATG9A 871:900 co-transfected wild-type ATG9A 871:900 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 6 5 theme ATG9A 919:923 arg1 oligomerization 925:939 ATG9A oligomerization 919:939 ATG9A oligomerization 919:939 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 4 6 theme amino 505:509 arg1 acids 511:515 amino acids L(340) to L(354) 505:532 amino acids L(340) to L(354) 505:532 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 7 7 theme mutation 1050:1057 arg1 consequences 1022:1033 the consequences 1018:1033 the consequences of the LYM/AAA mutation 1018:1057 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 3 8 theme molecular 353:361 arg1 determinants 363:374 molecular determinants 353:374 molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations 353:490 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 5 9 from region 696:701 arg1 located 670:676 located 670:676 located 670:676 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 8 10 theme protein 1427:1433 arg1 sorting 1411:1417 the sorting 1407:1417 the sorting of this protein 1407:1433 Taken together, these findings provide new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein. 27316455 8 11 theme intracellular 1269:1281 arg1 pathways 1283:1290 the intracellular pathways 1265:1290 the intracellular pathways followed by ATG9A to reach different subcellular compartments 1265:1352 Taken together, these findings provide new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein. 27316455 4 12 from retention 550:558 arg1 reticulum 588:596 the endoplasmic reticulum 572:596 the endoplasmic reticulum 572:596 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 7 13 theme intracellular 1066:1078 arg1 trafficking 1080:1090 the intracellular trafficking 1062:1090 the intracellular trafficking of ATG9A 1062:1099 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 6 14 theme biosynthetic 970:981 arg1 compartments 983:994 biosynthetic compartments 970:994 biosynthetic compartments 970:994 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 2 15 theme trans-Golgi 281:291 arg1 network 293:299 the trans-Golgi network 277:299 the trans-Golgi network 277:299 Under basal conditions, neosynthesized ATG9A proteins travel to the Golgi apparatus and cycle between the trans-Golgi network and endosomes. 27316455 5 16 theme sequence 660:667 arg1 substitution 626:637 substitution 626:637 substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues 626:731 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 5 17 theme C-terminal 685:694 arg1 region 696:701 the C-terminal region 681:701 the C-terminal region of ATG9A 681:710 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 7 18 theme synthesized 1129:1139 arg1 ATG9A 1141:1145 some newly synthesized ATG9A 1118:1145 some newly synthesized ATG9A 1118:1145 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 0 19 theme Molecular 0:8 arg1 determinants 10:21 Molecular determinants 0:21 Molecular determinants that mediate the sorting of human ATG9A from the endoplasmic reticulum 0:92 Molecular determinants that mediate the sorting of human ATG9A from the endoplasmic reticulum. 27316455 5 20 theme Golgi 777:781 arg1 apparatus 783:791 the Golgi apparatus 773:791 the Golgi apparatus 773:791 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 3 21 theme subcellular 392:402 arg1 trafficking 404:414 the subcellular trafficking 388:414 the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations 388:490 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 7 22 from study 1009:1013 arg1 trafficking 1080:1090 the intracellular trafficking 1062:1090 the intracellular trafficking of ATG9A 1062:1099 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 5 23 from located 670:676 arg1 region 696:701 the C-terminal region 681:701 the C-terminal region of ATG9A 681:710 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 3 24 theme HeLa 434:437 arg1 cells 439:443 HeLa cells 434:443 HeLa cells using sequential deletions and point mutations 434:490 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 5 25 theme alanine 716:722 arg1 residues 724:731 alanine residues 716:731 alanine residues 716:731 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 6 26 theme wild-type 886:894 arg1 ATG9A 896:900 co-transfected wild-type ATG9A 871:900 co-transfected wild-type ATG9A 871:900 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 8 27 theme subcellular 1329:1339 arg1 compartments 1341:1352 different subcellular compartments 1319:1352 different subcellular compartments 1319:1352 Taken together, these findings provide new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein. 27316455 7 28 theme ATG9A 1095:1099 arg1 trafficking 1080:1090 the intracellular trafficking 1062:1090 the intracellular trafficking of ATG9A 1062:1099 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 6 29 theme co-transfected 871:884 arg1 ATG9A 896:900 co-transfected wild-type ATG9A 871:900 co-transfected wild-type ATG9A 871:900 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 3 30 theme point 476:480 arg1 mutations 482:490 point mutations 476:490 point mutations 476:490 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 3 31 theme human 419:423 arg1 ATG9A 425:429 human ATG9A 419:429 human ATG9A 419:429 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 2 32 theme basal 181:185 arg1 conditions 187:196 basal conditions 181:196 basal conditions 181:196 Under basal conditions, neosynthesized ATG9A proteins travel to the Golgi apparatus and cycle between the trans-Golgi network and endosomes. 27316455 2 33 theme Golgi 243:247 arg1 apparatus 249:257 the Golgi apparatus 239:257 the Golgi apparatus 239:257 Under basal conditions, neosynthesized ATG9A proteins travel to the Golgi apparatus and cycle between the trans-Golgi network and endosomes. 27316455 3 34 theme sequential 451:460 arg1 deletions 462:470 sequential deletions 451:470 sequential deletions 451:470 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 3 35 theme ATG9A 425:429 arg1 trafficking 404:414 the subcellular trafficking 388:414 the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations 388:490 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 6 36 theme protein 858:864 arg1 oligomerization 828:842 oligomerization 828:842 oligomerization of the mutant protein with co-transfected wild-type ATG9A 828:900 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 5 37 theme YM 652:653 arg1 sequence 660:667 the L(711)YM(713) sequence 642:667 the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) 642:711 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 8 38 theme different 1319:1327 arg1 compartments 1341:1352 different subcellular compartments 1319:1352 different subcellular compartments 1319:1352 Taken together, these findings provide new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein. 27316455 0 39 theme human 51:55 arg1 ATG9A 57:61 human ATG9A 51:61 human ATG9A 51:61 Molecular determinants that mediate the sorting of human ATG9A from the endoplasmic reticulum. 27316455 4 40 dep L 517:517 arg1 to 524:525 to 524:525 to 524:525 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 4 40 dep L 517:517 arg1 L 527:527 L(354) 527:532 amino acids L(340) to L(354) 505:532 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 5 41 theme located 670:676 arg1 sequence 660:667 the L(711)YM(713) sequence 642:667 the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) 642:711 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 5 42 theme ATG9A 706:710 arg1 region 696:701 the C-terminal region 681:701 the C-terminal region of ATG9A 681:710 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 4 43 dep acids 511:515 arg1 L 517:517 L(340) 517:522 amino acids L(340) to L(354) 505:532 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 4 44 theme acids 511:515 arg1 Deletion 493:500 Deletion 493:500 Deletion of amino acids L(340) to L(354) 493:532 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 7 45 theme consequences 1022:1033 arg1 study 1009:1013 the study 1005:1013 the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A 1005:1099 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 1 46 theme multispanning 106:118 arg1 protein 129:135 a multispanning membrane protein 104:135 a multispanning membrane protein required for autophagosome formation 104:172 ATG9A is a multispanning membrane protein required for autophagosome formation. 27316455 1 46 theme multispanning 106:118 arg1 ATG9A 95:99 ATG9A 95:99 ATG9A 95:99 ATG9A is a multispanning membrane protein required for autophagosome formation. 27316455 2 47 theme ATG9A 214:218 arg1 proteins 220:227 neosynthesized ATG9A proteins 199:227 neosynthesized ATG9A proteins 199:227 Under basal conditions, neosynthesized ATG9A proteins travel to the Golgi apparatus and cycle between the trans-Golgi network and endosomes. 27316455 1 48 theme membrane 120:127 arg1 protein 129:135 a multispanning membrane protein 104:135 a multispanning membrane protein required for autophagosome formation 104:172 ATG9A is a multispanning membrane protein required for autophagosome formation. 27316455 1 48 theme membrane 120:127 arg1 ATG9A 95:99 ATG9A 95:99 ATG9A 95:99 ATG9A is a multispanning membrane protein required for autophagosome formation. 27316455 5 49 theme L 646:646 arg1 sequence 660:667 the L(711)YM(713) sequence 642:667 the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) 642:711 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 27316455 2 50 theme neosynthesized 199:212 arg1 proteins 220:227 neosynthesized ATG9A proteins 199:227 neosynthesized ATG9A proteins 199:227 Under basal conditions, neosynthesized ATG9A proteins travel to the Golgi apparatus and cycle between the trans-Golgi network and endosomes. 27316455 4 51 theme ATG9A 563:567 arg1 retention 550:558 the retention 546:558 the retention of ATG9A in the endoplasmic reticulum 546:596 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 0 52 from reticulum 84:92 arg1 sorting 40:46 the sorting 36:46 the sorting of human ATG9A from the endoplasmic reticulum 36:92 Molecular determinants that mediate the sorting of human ATG9A from the endoplasmic reticulum. 27316455 8 53 theme intramolecular 1368:1381 arg1 determinants 1383:1394 the intramolecular determinants 1364:1394 the intramolecular determinants that drive the sorting of this protein 1364:1433 Taken together, these findings provide new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein. 27316455 8 54 theme new 1247:1249 arg1 insights 1251:1258 new insights 1247:1258 new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein 1247:1433 Taken together, these findings provide new insights into the intracellular pathways followed by ATG9A to reach different subcellular compartments, and into the intramolecular determinants that drive the sorting of this protein. 27316455 7 55 theme Golgi 1162:1166 arg1 apparatus 1168:1176 the Golgi apparatus 1158:1176 the Golgi apparatus to reach the plasma membrane 1158:1205 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 4 56 theme endoplasmic 576:586 arg1 reticulum 588:596 the endoplasmic reticulum 572:596 the endoplasmic reticulum 572:596 Deletion of amino acids L(340) to L(354) resulted in the retention of ATG9A in the endoplasmic reticulum. 27316455 0 57 theme ATG9A 57:61 arg1 sorting 40:46 the sorting 36:46 the sorting of human ATG9A from the endoplasmic reticulum 36:92 Molecular determinants that mediate the sorting of human ATG9A from the endoplasmic reticulum. 27316455 3 58 from trafficking 404:414 arg1 cells 439:443 HeLa cells 434:443 HeLa cells using sequential deletions and point mutations 434:490 In the present work, we searched for molecular determinants involved in the subcellular trafficking of human ATG9A in HeLa cells using sequential deletions and point mutations. 27316455 7 59 theme plasma 1191:1196 arg1 membrane 1198:1205 the plasma membrane 1187:1205 the plasma membrane 1187:1205 Lastly, the study of the consequences of the LYM/AAA mutation on the intracellular trafficking of ATG9A highlighted that some newly synthesized ATG9A can bypass the Golgi apparatus to reach the plasma membrane. 27316455 6 60 theme mutant 851:856 arg1 protein 858:864 the mutant protein 847:864 the mutant protein 847:864 This defect could be corrected by oligomerization of the mutant protein with co-transfected wild-type ATG9A, suggesting that ATG9A oligomerization may help its sorting through biosynthetic compartments. 27316455 5 61 theme 713 655:657 arg1 sequence 660:667 the L(711)YM(713) sequence 642:667 the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) 642:711 In addition, we found that substitution of the L(711)YM(713) sequence (located in the C-terminal region of ATG9A) by alanine residues severely impaired its transport through the Golgi apparatus. 16940348 6 0 theme endosomal 1049:1057 arg1 membranes 1059:1067 peripheral, endosomal membranes 1037:1067 membranes 1059:1067 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 9 1 theme starvation-induced 1386:1403 arg1 autophagy 1405:1413 starvation-induced autophagy 1386:1413 starvation-induced autophagy 1386:1413 We speculate that starvation-induced autophagy, which requires mAtg9, may rely on an alteration of the steady-state trafficking of mAtg9, in a Atg1-dependent manner. 16940348 7 2 theme Atg1p 1192:1196 arg1 homologue 1179:1187 the putative mammalian homologue 1156:1187 the putative mammalian homologue of Atg1p 1156:1196 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 7 2 theme Atg1p 1192:1196 arg1 ULK1 1199:1202 ULK1 1199:1202 ULK1 1199:1202 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 3 3 from yeast 511:515 arg1 essential 498:506 essential 498:506 essential 498:506 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 5 4 theme cation-independent 849:866 arg1 receptor 888:895 the cation-independent mannose-6-phosphate receptor 845:895 the cation-independent mannose-6-phosphate receptor 845:895 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 4 theme cation-independent 849:866 arg1 TGN46 838:842 TGN46 838:842 TGN46 838:842 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 4 theme cation-independent 849:866 arg1 Rab9 907:910 Rab9 907:910 Rab9 907:910 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 4 theme cation-independent 849:866 arg1 Rab7 898:901 Rab7 898:901 Rab7 898:901 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 7 5 theme mammalian 1169:1177 arg1 homologue 1179:1187 the putative mammalian homologue 1156:1187 the putative mammalian homologue of Atg1p 1156:1196 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 7 5 theme mammalian 1169:1177 arg1 ULK1 1199:1202 ULK1 1199:1202 ULK1 1199:1202 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 9 6 theme mAtg9 1499:1503 arg1 trafficking 1484:1494 the steady-state trafficking 1467:1494 the steady-state trafficking of mAtg9 1467:1503 We speculate that starvation-induced autophagy, which requires mAtg9, may rely on an alteration of the steady-state trafficking of mAtg9, in a Atg1-dependent manner. 16940348 7 7 theme homologue 1179:1187 arg1 depletion 1143:1151 siRNA-mediated depletion 1128:1151 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, 1128:1203 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 6 8 theme marker 1112:1117 arg1 GFP-LC3 1119:1125 the autophagosomal marker GFP-LC3 1093:1125 the autophagosomal marker GFP-LC3 1093:1125 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 4 9 theme membrane 721:728 arg1 times 734:738 the membrane six times 717:738 the membrane six times 717:738 We show that both the N- and C-termini of mammalian Atg9 (mAtg9) are cytosolic, and predict that mAtg9 spans the membrane six times. 16940348 3 10 theme better 542:547 arg1 understanding 549:561 a better understanding 540:561 a better understanding of the function of this ubiquitious protein 540:605 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 9 11 theme trafficking 1484:1494 arg1 alteration 1453:1462 an alteration 1450:1462 an alteration of the steady-state trafficking of mAtg9 1450:1503 We speculate that starvation-induced autophagy, which requires mAtg9, may rely on an alteration of the steady-state trafficking of mAtg9, in a Atg1-dependent manner. 16940348 5 12 theme mannose-6-phosphate 868:886 arg1 receptor 888:895 the cation-independent mannose-6-phosphate receptor 845:895 the cation-independent mannose-6-phosphate receptor 845:895 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 12 theme mannose-6-phosphate 868:886 arg1 TGN46 838:842 TGN46 838:842 TGN46 838:842 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 12 theme mannose-6-phosphate 868:886 arg1 Rab9 907:910 Rab9 907:910 Rab9 907:910 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 12 theme mannose-6-phosphate 868:886 arg1 Rab7 898:901 Rab7 898:901 Rab7 898:901 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 3 13 theme multi-spanning 461:474 arg1 homologue 439:447 the mammalian homologue 425:447 the mammalian homologue of Atg9p 425:456 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 13 theme multi-spanning 461:474 arg1 protein 490:496 a multi-spanning transmembrane protein 459:496 a multi-spanning transmembrane protein essential in yeast for autophagy 459:529 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 6 14 theme mAtg9 1015:1019 arg1 redistribution 997:1010 a redistribution 995:1010 a redistribution 995:1010 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 6 14 theme mAtg9 1015:1019 arg1 positive 1080:1087 positive 1080:1087 positive 1080:1087 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 7 15 theme putative 1160:1167 arg1 homologue 1179:1187 the putative mammalian homologue 1156:1187 the putative mammalian homologue of Atg1p 1156:1196 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 7 15 theme putative 1160:1167 arg1 ULK1 1199:1202 ULK1 1199:1202 ULK1 1199:1202 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 1 16 theme normal 171:176 arg1 growth 178:183 normal growth 171:183 normal growth 171:183 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 5 17 from located 763:769 arg1 network 790:796 the trans-Golgi network 774:796 the trans-Golgi network 774:796 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 17 from located 763:769 arg1 endosomes 807:815 late endosomes 802:815 late endosomes 802:815 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 4 18 theme Atg9 660:663 arg1 N- 630:631 N- 630:631 N- 630:631 We show that both the N- and C-termini of mammalian Atg9 (mAtg9) are cytosolic, and predict that mAtg9 spans the membrane six times. 16940348 6 19 theme rapamycin 938:946 arg1 treatment 948:956 rapamycin treatment 938:956 rapamycin treatment 938:956 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 3 20 from essential 498:506 arg1 yeast 511:515 yeast 511:515 yeast 511:515 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 21 theme function 570:577 arg1 understanding 549:561 a better understanding 540:561 a better understanding of the function of this ubiquitious protein 540:605 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 22 theme mammalian 429:437 arg1 homologue 439:447 the mammalian homologue 425:447 the mammalian homologue of Atg9p 425:456 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 22 theme mammalian 429:437 arg1 protein 490:496 a multi-spanning transmembrane protein 459:496 a multi-spanning transmembrane protein essential in yeast for autophagy 459:529 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 0 23 dep TGN 68:70 arg1 the 64:66 the 64:66 the 64:66 Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. 16940348 5 24 located located 763:769 arg2 mAtg9 754:758 mAtg9 754:758 mAtg9 754:758 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 24 located located 763:769 arg1 network 790:796 the trans-Golgi network 774:796 the trans-Golgi network 774:796 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 5 24 located located 763:769 arg1 endosomes 807:815 late endosomes 802:815 late endosomes 802:815 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 1 25 theme certain 189:195 arg1 conditions 210:219 certain pathological conditions 189:219 certain pathological conditions 189:219 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 0 26 theme ULK1-dependent 15:28 arg1 cycling 30:36 ULK1-dependent cycling 15:36 ULK1-dependent cycling 15:36 Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. 16940348 9 27 theme steady-state 1471:1482 arg1 trafficking 1484:1494 the steady-state trafficking 1467:1494 the steady-state trafficking of mAtg9 1467:1503 We speculate that starvation-induced autophagy, which requires mAtg9, may rely on an alteration of the steady-state trafficking of mAtg9, in a Atg1-dependent manner. 16940348 8 28 theme amino 1355:1359 arg1 acids 1361:1365 amino acids 1355:1365 amino acids 1355:1365 The redistribution of mAtg9 also requires PI 3-kinase activity, and is reversed after restoration of amino acids. 16940348 1 29 theme pathological 197:208 arg1 conditions 210:219 certain pathological conditions 189:219 certain pathological conditions 189:219 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 0 30 theme mammalian 41:49 arg1 Atg9 51:54 mammalian Atg9 41:54 mammalian Atg9 41:54 Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. 16940348 6 31 theme acid 919:922 arg1 starvation 924:933 Amino acid starvation 913:933 Amino acid starvation 913:933 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 3 32 theme ubiquitious 587:597 arg1 protein 599:605 this ubiquitious protein 582:605 this ubiquitious protein 582:605 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 2 33 theme genes 379:383 arg1 identification 356:369 the identification 352:369 the identification of many genes required for autophagy 352:406 Autophagosomes are formed through a mechanism that is not well understood, despite the identification of many genes required for autophagy. 16940348 3 34 theme Atg9p 452:456 arg1 homologue 439:447 the mammalian homologue 425:447 the mammalian homologue of Atg9p 425:456 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 34 theme Atg9p 452:456 arg1 protein 490:496 a multi-spanning transmembrane protein 459:496 a multi-spanning transmembrane protein essential in yeast for autophagy 459:529 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 6 35 theme peripheral 1037:1046 arg1 membranes 1059:1067 peripheral, endosomal membranes 1037:1067 membranes 1059:1067 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 6 36 theme Amino 913:917 arg1 starvation 924:933 Amino acid starvation 913:933 Amino acid starvation 913:933 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 3 37 theme protein 599:605 arg1 function 570:577 the function 566:577 the function of this ubiquitious protein 566:605 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 5 38 theme late 802:805 arg1 endosomes 807:815 late endosomes 802:815 late endosomes 802:815 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 8 39 theme mAtg9 1276:1280 arg1 redistribution 1258:1271 The redistribution 1254:1271 The redistribution of mAtg9 1254:1280 The redistribution of mAtg9 also requires PI 3-kinase activity, and is reversed after restoration of amino acids. 16940348 3 40 theme essential 498:506 arg1 homologue 439:447 the mammalian homologue 425:447 the mammalian homologue of Atg9p 425:456 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 40 theme essential 498:506 arg1 protein 490:496 a multi-spanning transmembrane protein 459:496 a multi-spanning transmembrane protein essential in yeast for autophagy 459:529 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 0 41 theme Atg9 51:54 arg1 Starvation 0:9 Starvation 0:9 Starvation 0:9 Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. 16940348 0 41 theme Atg9 51:54 arg1 cycling 30:36 ULK1-dependent cycling 15:36 ULK1-dependent cycling 15:36 Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. 16940348 1 42 theme lysosomal 114:122 arg1 Autophagy 87:95 Autophagy 87:95 Autophagy 87:95 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 1 42 theme lysosomal 114:122 arg1 pathway 136:142 fundamentally a lysosomal degradation pathway 98:142 fundamentally a lysosomal degradation pathway 98:142 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 6 43 from TGN 1030:1032 arg1 redistribution 997:1010 a redistribution 995:1010 a redistribution 995:1010 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 6 43 from TGN 1030:1032 arg1 positive 1080:1087 positive 1080:1087 positive 1080:1087 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 4 44 theme mammalian 650:658 arg1 mAtg9 666:670 mAtg9 666:670 mAtg9 666:670 We show that both the N- and C-termini of mammalian Atg9 (mAtg9) are cytosolic, and predict that mAtg9 spans the membrane six times. 16940348 4 44 theme mammalian 650:658 arg1 Atg9 660:663 mammalian Atg9 650:663 mammalian Atg9 (mAtg9) 650:671 We show that both the N- and C-termini of mammalian Atg9 (mAtg9) are cytosolic, and predict that mAtg9 spans the membrane six times. 16940348 4 45 dep N- 630:631 arg1 the 626:628 the 626:628 the 626:628 We show that both the N- and C-termini of mammalian Atg9 (mAtg9) are cytosolic, and predict that mAtg9 spans the membrane six times. 16940348 1 46 theme degradation 124:134 arg1 Autophagy 87:95 Autophagy 87:95 Autophagy 87:95 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 1 46 theme degradation 124:134 arg1 pathway 136:142 fundamentally a lysosomal degradation pathway 98:142 fundamentally a lysosomal degradation pathway 98:142 Autophagy, fundamentally a lysosomal degradation pathway, functions in cells during normal growth and certain pathological conditions, including starvation, to maintain homeostasis. 16940348 6 47 theme autophagosomal 1097:1110 arg1 GFP-LC3 1119:1125 the autophagosomal marker GFP-LC3 1093:1125 the autophagosomal marker GFP-LC3 1093:1125 Amino acid starvation or rapamycin treatment, which upregulates autophagy, causes a redistribution of mAtg9 from the TGN to peripheral, endosomal membranes, which are positive for the autophagosomal marker GFP-LC3. 16940348 5 48 from network 790:796 arg1 located 763:769 located 763:769 located 763:769 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 8 49 theme 3-kinase 1299:1306 arg1 activity 1308:1315 PI 3-kinase activity 1296:1315 PI 3-kinase activity 1296:1315 The redistribution of mAtg9 also requires PI 3-kinase activity, and is reversed after restoration of amino acids. 16940348 8 50 theme PI 1296:1297 arg1 3-kinase 1299:1306 PI 3-kinase 1296:1306 PI 3-kinase activity 1296:1315 The redistribution of mAtg9 also requires PI 3-kinase activity, and is reversed after restoration of amino acids. 16940348 3 51 theme transmembrane 476:488 arg1 homologue 439:447 the mammalian homologue 425:447 the mammalian homologue of Atg9p 425:456 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 3 51 theme transmembrane 476:488 arg1 protein 490:496 a multi-spanning transmembrane protein 459:496 a multi-spanning transmembrane protein essential in yeast for autophagy 459:529 We have studied the mammalian homologue of Atg9p, a multi-spanning transmembrane protein essential in yeast for autophagy, to gain a better understanding of the function of this ubiquitious protein. 16940348 8 52 theme acids 1361:1365 arg1 restoration 1340:1350 restoration 1340:1350 restoration of amino acids 1340:1365 The redistribution of mAtg9 also requires PI 3-kinase activity, and is reversed after restoration of amino acids. 16940348 5 53 theme trans-Golgi 778:788 arg1 network 790:796 the trans-Golgi network 774:796 the trans-Golgi network 774:796 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 2 54 theme many 374:377 arg1 genes 379:383 many genes 374:383 many genes required for autophagy 374:406 Autophagosomes are formed through a mechanism that is not well understood, despite the identification of many genes required for autophagy. 16940348 9 55 theme Atg1-dependent 1511:1524 arg1 manner 1526:1531 a Atg1-dependent manner 1509:1531 a Atg1-dependent manner 1509:1531 We speculate that starvation-induced autophagy, which requires mAtg9, may rely on an alteration of the steady-state trafficking of mAtg9, in a Atg1-dependent manner. 16940348 7 56 theme siRNA-mediated 1128:1141 arg1 depletion 1143:1151 siRNA-mediated depletion 1128:1151 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, 1128:1203 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 16940348 5 57 from endosomes 807:815 arg1 located 763:769 located 763:769 located 763:769 We find that mAtg9 is located in the trans-Golgi network and late endosomes and colocalizes with TGN46, the cation-independent mannose-6-phosphate receptor, Rab7 and Rab9. 16940348 7 58 theme starvation-induced 1219:1236 arg1 redistribution 1238:1251 this starvation-induced redistribution 1214:1251 this starvation-induced redistribution 1214:1251 siRNA-mediated depletion of the putative mammalian homologue of Atg1p, ULK1, inhibits this starvation-induced redistribution. 9539703 2 0 theme leukocyte 324:332 arg1 LFA-1 363:367 LFA-1 363:367 LFA-1 363:367 ICAM-1 binds to leukocyte function-associated antigen (LFA-1) or macrophage-1 antigen (Mac-1). 9539703 2 0 theme leukocyte 324:332 arg1 antigen 354:360 leukocyte function-associated antigen 324:360 leukocyte function-associated antigen (LFA-1) 324:368 ICAM-1 binds to leukocyte function-associated antigen (LFA-1) or macrophage-1 antigen (Mac-1). 9539703 9 1 theme known 1471:1475 arg1 structure 1477:1485 the known structure 1467:1485 the known structure of the I-domain 1467:1501 Domain D1 has been docked with the known structure of the I-domain. 9539703 6 2 from different 962:970 arg1 structure 975:983 structure 975:983 structure to those of human ICAM-2 or murine ICAM-1, which do not bind rhinoviruses 975:1057 The loops are considerably different in structure to those of human ICAM-2 or murine ICAM-1, which do not bind rhinoviruses. 9539703 3 3 theme major 453:457 arg1 group 459:463 the major group 449:463 the major group of human rhinoviruses 449:485 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 3 3 theme major 453:457 arg1 rhinoviruses 474:485 human rhinoviruses 468:485 human rhinoviruses 468:485 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 7 4 theme charge 1080:1085 arg1 interactions 1087:1098 extensive charge interactions 1070:1098 extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses 1070:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 4 5 theme complex 774:780 arg1 reconstruction 736:749 a cryoelectron microscopy reconstruction 710:749 a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex 710:780 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 5 6 theme domain 879:884 arg1 DE 832:833 DE 832:833 DE 832:833 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 6 theme domain 879:884 arg1 loops 843:847 FG loops 840:847 FG loops 840:847 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 6 theme domain 879:884 arg1 CD 828:829 CD 828:829 CD 828:829 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 6 theme domain 879:884 arg1 BC 824:825 the BC 820:825 the BC 820:825 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 8 7 from -domain 1330:1336 arg1 chain 1351:1355 the alpha chain 1341:1355 the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs 1341:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 8 from group 1383:1387 arg1 ICAMs 1429:1433 ICAMs 1429:1433 ICAMs 1429:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 6 9 from structure 975:983 arg1 loops 939:943 The loops 935:943 The loops 935:943 The loops are considerably different in structure to those of human ICAM-2 or murine ICAM-1, which do not bind rhinoviruses. 9539703 6 9 from structure 975:983 arg1 different 962:970 different 962:970 different 962:970 The loops are considerably different in structure to those of human ICAM-2 or murine ICAM-1, which do not bind rhinoviruses. 9539703 10 10 theme structural 1574:1583 arg1 framework 1585:1593 a structural framework 1572:1593 a structural framework for the adhesion between these molecules 1572:1634 The resultant model is consistent with mutational data and provides a structural framework for the adhesion between these molecules. 9539703 5 11 from end 898:900 arg1 domain 879:884 the amino-terminal Ig-like domain 852:884 the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane 852:932 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 11 from end 898:900 arg1 loops 843:847 FG loops 840:847 FG loops 840:847 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 11 from end 898:900 arg1 D1 887:888 D1 887:888 D1 887:888 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 11 from end 898:900 arg1 CD 828:829 CD 828:829 CD 828:829 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 11 from end 898:900 arg1 DE 832:833 DE 832:833 DE 832:833 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 11 from end 898:900 arg1 BC 824:825 the BC 820:825 the BC 820:825 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 10 12 theme resultant 1508:1516 arg1 consistent 1527:1536 consistent 1527:1536 consistent 1527:1536 The resultant model is consistent with mutational data and provides a structural framework for the adhesion between these molecules. 9539703 10 12 theme resultant 1508:1516 arg1 model 1518:1522 The resultant model 1504:1522 The resultant model 1504:1522 The resultant model is consistent with mutational data and provides a structural framework for the adhesion between these molecules. 9539703 8 13 theme LFA-1 1360:1364 arg1 chain 1351:1355 the alpha chain 1341:1355 the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs 1341:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 5 14 theme Rhinovirus 783:792 arg1 attachment 794:803 Rhinovirus attachment 783:803 Rhinovirus attachment 783:803 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 4 15 theme three-dimensional 563:579 arg1 structure 588:596 The three-dimensional atomic structure 559:596 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 559:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 3 16 theme human 468:472 arg1 rhinoviruses 474:485 human rhinoviruses 468:485 human rhinoviruses 468:485 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 4 17 theme domains 624:630 arg1 structure 588:596 The three-dimensional atomic structure 559:596 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 559:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 7 18 theme rhinoviruses 1209:1220 arg1 rhinoviruses 1209:1220 rhinoviruses 1209:1220 rhinoviruses 1209:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 7 18 theme rhinoviruses 1209:1220 arg1 groups 1199:1204 both major and minor receptor groups 1169:1204 both major and minor receptor groups of rhinoviruses 1169:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 7 19 theme extensive 1070:1078 arg1 interactions 1087:1098 extensive charge interactions 1070:1098 extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses 1070:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 8 20 theme carboxyl 1374:1381 arg1 group 1383:1387 the carboxyl group 1370:1387 the carboxyl group of a conserved glutamic acid residue on ICAMs 1370:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 20 theme carboxyl 1374:1381 arg1 residue 1418:1424 a conserved glutamic acid residue 1392:1424 a conserved glutamic acid residue on ICAMs 1392:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 21 from residue 1418:1424 arg1 ICAMs 1429:1433 ICAMs 1429:1433 ICAMs 1429:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 7 22 theme receptor 1190:1197 arg1 rhinoviruses 1209:1220 rhinoviruses 1209:1220 rhinoviruses 1209:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 7 22 theme receptor 1190:1197 arg1 groups 1199:1204 both major and minor receptor groups 1169:1204 both major and minor receptor groups of rhinoviruses 1169:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 3 23 theme rhinoviruses 474:485 arg1 group 459:463 the major group 449:463 the major group of human rhinoviruses 449:485 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 3 23 theme rhinoviruses 474:485 arg1 rhinoviruses 474:485 human rhinoviruses 468:485 human rhinoviruses 468:485 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 8 24 theme group 1383:1387 arg1 chain 1351:1355 the alpha chain 1341:1355 the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs 1341:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 5 25 from loops 843:847 arg1 end 898:900 the end 894:900 the end 894:900 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 26 theme amino-terminal 856:869 arg1 D1 887:888 D1 887:888 D1 887:888 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 26 theme amino-terminal 856:869 arg1 domain 879:884 the amino-terminal Ig-like domain 852:884 the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane 852:932 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 1 27 theme normal 152:157 arg1 function 159:166 The normal function 148:166 The normal function of human intercellular adhesion molecule-1 (ICAM-1) 148:218 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 8 28 theme ICAMs 1242:1246 arg1 interaction 1227:1237 The interaction 1223:1237 The interaction of ICAMs with LFA-1 1223:1257 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 4 29 theme amino-terminal 609:622 arg1 D2 640:641 D2 640:641 D2 640:641 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 29 theme amino-terminal 609:622 arg1 ICAM-1 647:652 ICAM-1 647:652 ICAM-1 647:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 29 theme amino-terminal 609:622 arg1 D1 633:634 D1 633:634 D1 633:634 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 29 theme amino-terminal 609:622 arg1 domains 624:630 the two amino-terminal domains 601:630 the two amino-terminal domains (D1 and D2) of ICAM-1 601:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 1 30 theme endothelial 251:261 arg1 cells 263:267 endothelial cells 251:267 endothelial cells 251:267 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 8 31 theme insertion 1317:1325 arg1 -domain 1330:1336 the insertion (I)-domain 1313:1336 the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs 1313:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 4 32 theme rhinovirus-ICAM-1 756:772 arg1 complex 774:780 a rhinovirus-ICAM-1 complex 754:780 a rhinovirus-ICAM-1 complex 754:780 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 3 33 used used 427:430 arg2 ICAM-1 412:417 ICAM-1 412:417 ICAM-1 412:417 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 3 33 used used 427:430 arg2 receptor 437:444 a receptor 435:444 a receptor 435:444 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 4 34 theme atomic 581:586 arg1 structure 588:596 The three-dimensional atomic structure 559:596 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 559:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 8 35 theme conserved 1394:1402 arg1 residue 1418:1424 a conserved glutamic acid residue 1392:1424 a conserved glutamic acid residue on ICAMs 1392:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 0 36 theme rhinovirus 95:104 arg1 receptor 106:113 a rhinovirus receptor 93:113 a rhinovirus receptor 93:113 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 0 36 theme rhinovirus 95:104 arg1 it 77:78 it 77:78 it 77:78 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 5 37 theme Ig-like 871:877 arg1 D1 887:888 D1 887:888 D1 887:888 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 37 theme Ig-like 871:877 arg1 domain 879:884 the amino-terminal Ig-like domain 852:884 the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane 852:932 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 38 theme cellular 916:923 arg1 membrane 925:932 the cellular membrane 912:932 the cellular membrane 912:932 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 1 39 theme human 171:175 arg1 ICAM-1 212:217 ICAM-1 212:217 ICAM-1 212:217 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 1 39 theme human 171:175 arg1 molecule-1 200:209 human intercellular adhesion molecule-1 171:209 human intercellular adhesion molecule-1 (ICAM-1) 171:218 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 0 40 theme amino-terminal 25:38 arg1 domains 40:46 the two amino-terminal domains 17:46 the two amino-terminal domains of human ICAM-1 17:62 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 0 40 theme amino-terminal 25:38 arg1 ICAM-1 57:62 human ICAM-1 51:62 human ICAM-1 51:62 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 8 41 theme residue 1418:1424 arg1 group 1383:1387 the carboxyl group 1370:1387 the carboxyl group of a conserved glutamic acid residue on ICAMs 1370:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 41 theme residue 1418:1424 arg1 residue 1418:1424 a conserved glutamic acid residue 1392:1424 a conserved glutamic acid residue on ICAMs 1392:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 41 theme residue 1418:1424 arg1 LFA-1 1360:1364 LFA-1 1360:1364 LFA-1 1360:1364 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 7 42 theme major 1174:1178 arg1 rhinoviruses 1209:1220 rhinoviruses 1209:1220 rhinoviruses 1209:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 7 42 theme major 1174:1178 arg1 groups 1199:1204 both major and minor receptor groups 1169:1204 both major and minor receptor groups of rhinoviruses 1169:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 1 43 theme intercellular 177:189 arg1 ICAM-1 212:217 ICAM-1 212:217 ICAM-1 212:217 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 1 43 theme intercellular 177:189 arg1 molecule-1 200:209 human intercellular adhesion molecule-1 171:209 human intercellular adhesion molecule-1 (ICAM-1) 171:218 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 10 44 theme mutational 1543:1552 arg1 data 1554:1557 mutational data 1543:1557 mutational data 1543:1557 The resultant model is consistent with mutational data and provides a structural framework for the adhesion between these molecules. 9539703 8 45 theme acid 1413:1416 arg1 residue 1418:1424 a conserved glutamic acid residue 1392:1424 a conserved glutamic acid residue on ICAMs 1392:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 46 from ICAMs 1429:1433 arg1 group 1383:1387 the carboxyl group 1370:1387 the carboxyl group of a conserved glutamic acid residue on ICAMs 1370:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 46 from ICAMs 1429:1433 arg1 residue 1418:1424 a conserved glutamic acid residue 1392:1424 a conserved glutamic acid residue on ICAMs 1392:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 8 46 from ICAMs 1429:1433 arg1 LFA-1 1360:1364 LFA-1 1360:1364 LFA-1 1360:1364 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 10 47 with consistent 1527:1536 arg1 data 1554:1557 mutational data 1543:1557 mutational data 1543:1557 The resultant model is consistent with mutational data and provides a structural framework for the adhesion between these molecules. 9539703 1 48 theme adhesion 191:198 arg1 ICAM-1 212:217 ICAM-1 212:217 ICAM-1 212:217 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 1 48 theme adhesion 191:198 arg1 molecule-1 200:209 human intercellular adhesion molecule-1 171:209 human intercellular adhesion molecule-1 (ICAM-1) 171:218 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 5 49 from CD 828:829 arg1 end 898:900 the end 894:900 the end 894:900 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 8 50 theme glutamic 1404:1411 arg1 residue 1418:1424 a conserved glutamic acid residue 1392:1424 a conserved glutamic acid residue on ICAMs 1392:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 0 51 theme LFA-1 125:129 arg1 it 77:78 it 77:78 it 77:78 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 0 51 theme LFA-1 125:129 arg1 ligand 140:145 an LFA-1 integrin ligand 122:145 an LFA-1 integrin ligand 122:145 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 7 52 theme human 1119:1123 arg1 rhinoviruses 1125:1136 human rhinoviruses 1119:1136 human rhinoviruses 1119:1136 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 3 53 theme subsequent 513:522 arg1 uncoating 530:538 the subsequent viral uncoating 509:538 the subsequent viral uncoating during cell entry 509:556 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 0 54 theme domains 40:46 arg1 structure 4:12 The structure 0:12 The structure of the two amino-terminal domains of human ICAM-1 0:62 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 9 55 theme I-domain 1494:1501 arg1 structure 1477:1485 the known structure 1467:1485 the known structure of the I-domain 1467:1501 Domain D1 has been docked with the known structure of the I-domain. 9539703 3 56 theme viral 524:528 arg1 uncoating 530:538 the subsequent viral uncoating 509:538 the subsequent viral uncoating during cell entry 509:556 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 6 57 theme murine 1013:1018 arg1 ICAM-1 1020:1025 murine ICAM-1 1013:1025 murine ICAM-1 1013:1025 The loops are considerably different in structure to those of human ICAM-2 or murine ICAM-1, which do not bind rhinoviruses. 9539703 8 58 theme divalent 1288:1295 arg1 cation 1297:1302 a divalent cation 1286:1302 a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs 1286:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 5 59 theme distal 902:907 arg1 D1 887:888 D1 887:888 D1 887:888 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 59 theme distal 902:907 arg1 domain 879:884 the amino-terminal Ig-like domain 852:884 the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane 852:932 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 9 60 theme Domain 1436:1441 arg1 D1 1443:1444 Domain D1 1436:1444 Domain D1 1436:1444 Domain D1 has been docked with the known structure of the I-domain. 9539703 4 61 dep domains 624:630 arg1 D2 640:641 D2 640:641 D2 640:641 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 61 dep domains 624:630 arg1 ICAM-1 647:652 ICAM-1 647:652 ICAM-1 647:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 61 dep domains 624:630 arg1 D1 633:634 D1 633:634 D1 633:634 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 61 dep domains 624:630 arg1 domains 624:630 the two amino-terminal domains 601:630 the two amino-terminal domains (D1 and D2) of ICAM-1 601:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 62 theme cryoelectron 712:723 arg1 microscopy 725:734 cryoelectron microscopy 712:734 a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex 710:780 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 5 63 from DE 832:833 arg1 end 898:900 the end 894:900 the end 894:900 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 5 64 theme FG 840:841 arg1 loops 843:847 FG loops 840:847 FG loops 840:847 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 4 65 theme 2.2-A 677:681 arg1 resolution 683:692 2.2-A resolution 677:692 2.2-A resolution 677:692 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 1 66 theme molecule-1 200:209 arg1 function 159:166 The normal function 148:166 The normal function of human intercellular adhesion molecule-1 (ICAM-1) 148:218 The normal function of human intercellular adhesion molecule-1 (ICAM-1) is to provide adhesion between endothelial cells and leukocytes after injury or stress. 9539703 0 67 theme integrin 131:138 arg1 it 77:78 it 77:78 it 77:78 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 0 67 theme integrin 131:138 arg1 ligand 140:145 an LFA-1 integrin ligand 122:145 an LFA-1 integrin ligand 122:145 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 4 68 theme ICAM-1 647:652 arg1 D2 640:641 D2 640:641 D2 640:641 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 68 theme ICAM-1 647:652 arg1 ICAM-1 647:652 ICAM-1 647:652 ICAM-1 647:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 68 theme ICAM-1 647:652 arg1 D1 633:634 D1 633:634 D1 633:634 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 4 68 theme ICAM-1 647:652 arg1 domains 624:630 the two amino-terminal domains 601:630 the two amino-terminal domains (D1 and D2) of ICAM-1 601:652 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 0 69 theme ICAM-1 57:62 arg1 domains 40:46 the two amino-terminal domains 17:46 the two amino-terminal domains of human ICAM-1 17:62 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 0 69 theme ICAM-1 57:62 arg1 ICAM-1 57:62 human ICAM-1 51:62 human ICAM-1 51:62 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 2 70 theme macrophage-1 373:384 arg1 antigen 386:392 macrophage-1 antigen 373:392 macrophage-1 antigen (Mac-1) 373:400 ICAM-1 binds to leukocyte function-associated antigen (LFA-1) or macrophage-1 antigen (Mac-1). 9539703 2 70 theme macrophage-1 373:384 arg1 Mac-1 395:399 Mac-1 395:399 Mac-1 395:399 ICAM-1 binds to leukocyte function-associated antigen (LFA-1) or macrophage-1 antigen (Mac-1). 9539703 3 71 theme cell 547:550 arg1 entry 552:556 cell entry 547:556 cell entry 547:556 However, ICAM-1 is also used as a receptor by the major group of human rhinoviruses and is a catalyst for the subsequent viral uncoating during cell entry. 9539703 5 72 from BC 824:825 arg1 end 898:900 the end 894:900 the end 894:900 Rhinovirus attachment is confined to the BC, CD, DE, and FG loops of the amino-terminal Ig-like domain (D1) at the end distal to the cellular membrane. 9539703 6 73 theme human 997:1001 arg1 ICAM-2 1003:1008 human ICAM-2 997:1008 human ICAM-2 997:1008 The loops are considerably different in structure to those of human ICAM-2 or murine ICAM-1, which do not bind rhinoviruses. 9539703 7 74 theme minor 1184:1188 arg1 rhinoviruses 1209:1220 rhinoviruses 1209:1220 rhinoviruses 1209:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 7 74 theme minor 1184:1188 arg1 groups 1199:1204 both major and minor receptor groups 1169:1204 both major and minor receptor groups of rhinoviruses 1169:1220 There are extensive charge interactions between ICAM-1 and human rhinoviruses, which are mostly conserved in both major and minor receptor groups of rhinoviruses. 9539703 8 75 with interaction 1227:1237 arg1 LFA-1 1253:1257 LFA-1 1253:1257 LFA-1 1253:1257 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 0 76 theme human 51:55 arg1 ICAM-1 57:62 human ICAM-1 51:62 human ICAM-1 51:62 The structure of the two amino-terminal domains of human ICAM-1 suggests how it functions as a rhinovirus receptor and as an LFA-1 integrin ligand. 9539703 8 77 theme alpha 1345:1349 arg1 chain 1351:1355 the alpha chain 1341:1355 the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs 1341:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 9539703 4 78 theme microscopy 725:734 arg1 reconstruction 736:749 a cryoelectron microscopy reconstruction 710:749 a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex 710:780 The three-dimensional atomic structure of the two amino-terminal domains (D1 and D2) of ICAM-1 has been determined to 2.2-A resolution and fitted into a cryoelectron microscopy reconstruction of a rhinovirus-ICAM-1 complex. 9539703 2 79 theme function-associated 334:352 arg1 LFA-1 363:367 LFA-1 363:367 LFA-1 363:367 ICAM-1 binds to leukocyte function-associated antigen (LFA-1) or macrophage-1 antigen (Mac-1). 9539703 2 79 theme function-associated 334:352 arg1 antigen 354:360 leukocyte function-associated antigen 324:360 leukocyte function-associated antigen (LFA-1) 324:368 ICAM-1 binds to leukocyte function-associated antigen (LFA-1) or macrophage-1 antigen (Mac-1). 9539703 8 80 from LFA-1 1360:1364 arg1 ICAMs 1429:1433 ICAMs 1429:1433 ICAMs 1429:1433 The interaction of ICAMs with LFA-1 is known to be mediated by a divalent cation bound to the insertion (I)-domain on the alpha chain of LFA-1 and the carboxyl group of a conserved glutamic acid residue on ICAMs. 1898736 0 0 theme pre-alpha-inhibitor 83:101 arg1 chains 49:54 the chains 45:54 the chains of the human blood protein pre-alpha-inhibitor 45:101 Chondroitin 4-sulfate covalently cross-links the chains of the human blood protein pre-alpha-inhibitor. 1898736 2 1 theme PGP 360:362 arg1 cross-link 365:374 a protein-glycosaminoglycan-protein (PGP) cross-link 323:374 a protein-glycosaminoglycan-protein (PGP) cross-link 323:374 The chains are covalently linked to each other by a structure that has not previously been described, which we designate a protein-glycosaminoglycan-protein (PGP) cross-link. 1898736 0 2 theme protein 75:81 arg1 pre-alpha-inhibitor 83:101 the human blood protein pre-alpha-inhibitor 59:101 the human blood protein pre-alpha-inhibitor 59:101 Chondroitin 4-sulfate covalently cross-links the chains of the human blood protein pre-alpha-inhibitor. 1898736 5 3 from present 793:799 arg1 proteins 810:817 other proteins 804:817 other proteins 804:817 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 3 4 theme light 602:606 arg1 chain 608:612 the light chain 598:612 the light chain 598:612 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 3 5 theme O-glycosidic 567:578 arg1 link 580:583 a typical O-glycosidic link 557:583 a typical O-glycosidic link to Ser-10 of the light chain 557:612 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 4 6 theme chain 759:763 arg1 N-acetylgalactosamine 712:732 an internal N-acetylgalactosamine 700:732 an internal N-acetylgalactosamine of the glycosaminoglycan chain 700:763 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 2 7 attach linked 228:233 arg3 structure 254:262 a structure 252:262 a structure that has not previously been described, which we designate a protein-glycosaminoglycan-protein (PGP) cross-link 252:374 The chains are covalently linked to each other by a structure that has not previously been described, which we designate a protein-glycosaminoglycan-protein (PGP) cross-link. 1898736 2 7 attach linked 228:233 arg2 chains 206:211 The chains 202:211 The chains 202:211 The chains are covalently linked to each other by a structure that has not previously been described, which we designate a protein-glycosaminoglycan-protein (PGP) cross-link. 1898736 2 7 attach linked 228:233 arg1 other 243:247 other 243:247 other 243:247 The chains are covalently linked to each other by a structure that has not previously been described, which we designate a protein-glycosaminoglycan-protein (PGP) cross-link. 1898736 1 8 theme protein 120:126 arg1 pre-alpha-inhibitor 128:146 The human blood protein pre-alpha-inhibitor 104:146 The human blood protein pre-alpha-inhibitor 104:146 The human blood protein pre-alpha-inhibitor is composed of one heavy and one light protein chain. 1898736 3 9 dep protein 394:400 arg1 techniques 430:439 analytical techniques 419:439 analytical techniques 419:439 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 4 10 theme Asp 685:687 arg1 alpha-carbon 654:665 the alpha-carbon 650:665 the alpha-carbon of its C-terminal Asp 650:687 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 5 11 from proteins 810:817 arg1 present 793:799 present 793:799 present 793:799 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 5 12 attach present 793:799 arg2 cross-link 775:784 This PGP cross-link 766:784 This PGP cross-link 766:784 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 5 12 attach present 793:799 arg1 proteins 810:817 other proteins 804:817 other proteins 804:817 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 3 13 theme typical 559:565 arg1 link 580:583 a typical O-glycosidic link 557:583 a typical O-glycosidic link to Ser-10 of the light chain 557:612 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 1 14 theme light 181:185 arg1 chain 195:199 one heavy and one light protein chain 163:199 chain 195:199 The human blood protein pre-alpha-inhibitor is composed of one heavy and one light protein chain. 1898736 4 15 theme glycosaminoglycan 741:757 arg1 chain 759:763 the glycosaminoglycan chain 737:763 the glycosaminoglycan chain 737:763 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 1 16 theme protein 187:193 arg1 chain 195:199 one heavy and one light protein chain 163:199 chain 195:199 The human blood protein pre-alpha-inhibitor is composed of one heavy and one light protein chain. 1898736 0 17 theme Chondroitin 0:10 arg1 4-sulfate 12:20 Chondroitin 4-sulfate 0:20 Chondroitin 4-sulfate 0:20 Chondroitin 4-sulfate covalently cross-links the chains of the human blood protein pre-alpha-inhibitor. 1898736 4 18 theme C-terminal 674:683 arg1 Asp 685:687 its C-terminal Asp 670:687 its C-terminal Asp 670:687 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 1 19 theme human 108:112 arg1 pre-alpha-inhibitor 128:146 The human blood protein pre-alpha-inhibitor 104:146 The human blood protein pre-alpha-inhibitor 104:146 The human blood protein pre-alpha-inhibitor is composed of one heavy and one light protein chain. 1898736 1 20 theme blood 114:118 arg1 pre-alpha-inhibitor 128:146 The human blood protein pre-alpha-inhibitor 104:146 The human blood protein pre-alpha-inhibitor 104:146 The human blood protein pre-alpha-inhibitor is composed of one heavy and one light protein chain. 1898736 3 21 theme carbohydrate 406:417 arg1 combination 379:389 A combination 377:389 A combination of protein and carbohydrate analytical techniques 377:439 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 3 22 theme interchain 460:469 arg1 linkage 471:477 the interchain linkage 456:477 the interchain linkage 456:477 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 3 23 theme analytical 419:428 arg1 techniques 430:439 analytical techniques 419:439 analytical techniques 419:439 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 5 24 theme heterogeneous 862:874 arg1 behavior 876:883 the heterogeneous behavior 858:883 the heterogeneous behavior of proteins containing glycosaminoglycan 858:924 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 3 25 theme protein 394:400 arg1 combination 379:389 A combination 377:389 A combination of protein and carbohydrate analytical techniques 377:439 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 5 26 theme PGP 771:773 arg1 cross-link 775:784 This PGP cross-link 766:784 This PGP cross-link 766:784 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 2 27 theme protein-glycosaminoglycan-protein 325:357 arg1 cross-link 365:374 a protein-glycosaminoglycan-protein (PGP) cross-link 323:374 a protein-glycosaminoglycan-protein (PGP) cross-link 323:374 The chains are covalently linked to each other by a structure that has not previously been described, which we designate a protein-glycosaminoglycan-protein (PGP) cross-link. 1898736 3 28 theme chondroitin 496:506 arg1 glycosaminoglycan 518:534 a chondroitin 4-sulfate glycosaminoglycan 494:534 a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain 494:612 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 5 29 theme other 804:808 arg1 proteins 810:817 other proteins 804:817 other proteins 804:817 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 3 30 theme chain 608:612 arg1 Ser-10 588:593 Ser-10 588:593 Ser-10 of the light chain 588:612 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 3 31 theme 4-sulfate 508:516 arg1 glycosaminoglycan 518:534 a chondroitin 4-sulfate glycosaminoglycan 494:534 a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain 494:612 A combination of protein and carbohydrate analytical techniques indicates that the interchain linkage is mediated by a chondroitin 4-sulfate glycosaminoglycan that originates from a typical O-glycosidic link to Ser-10 of the light chain. 1898736 4 32 theme internal 703:710 arg1 N-acetylgalactosamine 712:732 an internal N-acetylgalactosamine 700:732 an internal N-acetylgalactosamine of the glycosaminoglycan chain 700:763 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 5 33 located present 793:799 arg2 cross-link 775:784 This PGP cross-link 766:784 This PGP cross-link 766:784 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 5 33 located present 793:799 arg1 proteins 810:817 other proteins 804:817 other proteins 804:817 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 4 34 theme N-acetylgalactosamine 712:732 arg1 C-6 693:695 C-6 693:695 C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain 693:763 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 4 35 theme heavy 619:623 arg1 chain 625:629 The heavy chain 615:629 The heavy chain 615:629 The heavy chain is esterified, via the alpha-carbon of its C-terminal Asp, to C-6 of an internal N-acetylgalactosamine of the glycosaminoglycan chain. 1898736 0 36 theme blood 69:73 arg1 pre-alpha-inhibitor 83:101 the human blood protein pre-alpha-inhibitor 59:101 the human blood protein pre-alpha-inhibitor 59:101 Chondroitin 4-sulfate covalently cross-links the chains of the human blood protein pre-alpha-inhibitor. 1898736 5 37 contain containing 897:906 arg2 glycosaminoglycan 908:924 glycosaminoglycan 908:924 glycosaminoglycan 908:924 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 5 37 contain containing 897:906 arg1 proteins 888:895 proteins 888:895 proteins containing glycosaminoglycan 888:924 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 1898736 0 38 theme human 63:67 arg1 pre-alpha-inhibitor 83:101 the human blood protein pre-alpha-inhibitor 59:101 the human blood protein pre-alpha-inhibitor 59:101 Chondroitin 4-sulfate covalently cross-links the chains of the human blood protein pre-alpha-inhibitor. 1898736 5 39 theme proteins 888:895 arg1 behavior 876:883 the heterogeneous behavior 858:883 the heterogeneous behavior of proteins containing glycosaminoglycan 858:924 This PGP cross-link may be present in other proteins, but could have been overlooked due to the heterogeneous behavior of proteins containing glycosaminoglycan. 11279095 1 0 theme neuronal 151:158 arg1 development 160:170 neuronal development 151:170 neuronal development 151:170 NCAM polysialylation plays a critical role in neuronal development and regeneration. 11279095 2 1 gly Polysialylation 190:204 arg1 NCAM 244:247 NCAM 244:247 NCAM 244:247 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 1 gly Polysialylation 190:204 arg1 molecule 234:241 the neural cell adhesion molecule 209:241 the neural cell adhesion molecule (NCAM) 209:248 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 11 2 theme intramolecular 1833:1846 arg1 bonds 1858:1862 intramolecular disulfide bonds 1833:1862 intramolecular disulfide bonds 1833:1862 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 3 3 theme COOH 601:604 arg1 terminus 606:613 the COOH terminus 597:613 the COOH terminus 597:613 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 11 4 theme ST8Sia 1982:1987 arg1 activity 1970:1977 the catalytic activity 1956:1977 the catalytic activity of ST8Sia IV 1956:1990 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 7 5 from terminus 1365:1372 arg1 cysteine 1310:1317 the second cysteine 1299:1317 the second cysteine in sialylmotif L 1299:1334 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 7 5 from terminus 1365:1372 arg1 cysteine 1344:1351 the cysteine 1340:1351 the cysteine at the COOH terminus 1340:1372 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 10 6 theme COOH 1700:1703 arg1 terminus 1705:1712 the COOH terminus 1696:1712 the COOH terminus of ST8Sia IV 1696:1725 By contrast, the addition of green fluorescence protein at the COOH terminus of ST8Sia IV did not render the enzyme inactive. 11279095 11 7 theme catalytic 1960:1968 arg1 activity 1970:1977 the catalytic activity 1956:1977 the catalytic activity of ST8Sia IV 1956:1990 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 10 8 theme green 1666:1670 arg1 protein 1685:1691 green fluorescence protein 1666:1691 green fluorescence protein 1666:1691 By contrast, the addition of green fluorescence protein at the COOH terminus of ST8Sia IV did not render the enzyme inactive. 11279095 5 9 theme Trichoplusia 856:867 arg1 cells 849:853 insect cells 842:853 insect cells 842:853 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 5 9 theme Trichoplusia 856:867 arg1 ni 869:870 Trichoplusia ni 856:870 Trichoplusia ni 856:870 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 4 10 theme disulfide 663:671 arg1 bonds 673:677 disulfide bonds 663:677 disulfide bonds 663:677 However, structural information, including how disulfide bonds are formed, has not been determined for any of the sialyltransferases. 11279095 7 11 theme disulfide 1388:1396 arg1 bridge 1398:1403 a second disulfide bridge 1379:1403 a second disulfide bridge 1379:1403 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 11 12 theme disulfide 1848:1856 arg1 bonds 1858:1862 intramolecular disulfide bonds 1833:1862 intramolecular disulfide bonds 1833:1862 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 10 13 from terminus 1705:1712 arg1 addition 1654:1661 the addition 1650:1661 the addition of green fluorescence protein at the COOH terminus of ST8Sia IV 1650:1725 By contrast, the addition of green fluorescence protein at the COOH terminus of ST8Sia IV did not render the enzyme inactive. 11279095 7 14 from cysteine 1206:1213 arg1 L 1230:1230 sialylmotif L 1218:1230 sialylmotif L 1218:1230 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 7 14 from cysteine 1206:1213 arg1 S 1264:1264 sialylmotif S 1252:1264 sialylmotif S 1252:1264 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 3 15 theme ST8Sia 437:442 arg1 family 449:454 the ST8Sia gene family 433:454 the ST8Sia gene family 433:454 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 4 16 theme structural 625:634 arg1 information 636:646 structural information 625:646 structural information 625:646 However, structural information, including how disulfide bonds are formed, has not been determined for any of the sialyltransferases. 11279095 7 17 theme disulfide 1273:1281 arg1 bridge 1283:1288 a disulfide bridge 1271:1288 a disulfide bridge 1271:1288 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 8 18 theme disulfide 1496:1504 arg1 bridges 1506:1512 the disulfide bridges 1492:1512 the disulfide bridges 1492:1512 Site-directed mutagenesis demonstrated that mutation at cysteine residues involved in the disulfide bridges completely inactivated the enzyme. 11279095 11 19 theme sterical 1804:1811 arg1 critical 1943:1950 critical 1943:1950 critical 1943:1950 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 11 19 theme sterical 1804:1811 arg1 structure 1813:1821 the sterical structure 1800:1821 the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, 1800:1938 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 0 20 located found 33:37 arg1 polysialyltransferase 52:72 ST8Sia IV polysialyltransferase 42:72 ST8Sia IV polysialyltransferase 42:72 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase are required for its activity. 11279095 0 20 located found 33:37 arg2 structures 22:31 Unique disulfide bond structures 0:31 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase 0:72 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase are required for its activity. 11279095 3 21 from cysteine 550:557 arg1 S 574:574 sialylmotif S 562:574 sialylmotif S 562:574 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 7 22 theme first 1200:1204 arg1 cysteine 1206:1213 the first cysteine 1196:1213 the first cysteine in sialylmotif L 1196:1230 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 0 23 theme disulfide 7:15 arg1 structures 22:31 Unique disulfide bond structures 0:31 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase 0:72 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase are required for its activity. 11279095 7 24 theme second 1303:1308 arg1 cysteine 1310:1317 the second cysteine 1299:1317 the second cysteine in sialylmotif L 1299:1334 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 11 25 theme COOH 1902:1905 arg1 terminus 1907:1914 the COOH terminus 1898:1914 the COOH terminus 1898:1914 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 0 26 theme Unique 0:5 arg1 structures 22:31 Unique disulfide bond structures 0:31 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase 0:72 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase are required for its activity. 11279095 5 27 theme insect 842:847 arg1 cells 849:853 insect cells 842:853 insect cells 842:853 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 5 27 theme insect 842:847 arg1 ni 869:870 Trichoplusia ni 856:870 Trichoplusia ni 856:870 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 3 28 from cysteines 518:526 arg1 cysteine 585:592 one cysteine 581:592 one cysteine at the COOH terminus 581:613 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 28 from cysteines 518:526 arg1 L 543:543 sialylmotif L 531:543 sialylmotif L 531:543 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 28 from cysteines 518:526 arg1 cysteine 550:557 one cysteine 546:557 one cysteine in sialylmotif S 546:574 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 28 from cysteines 518:526 arg1 terminus 606:613 the COOH terminus 597:613 the COOH terminus 597:613 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 7 29 from cysteine 1310:1317 arg1 L 1334:1334 sialylmotif L 1322:1334 sialylmotif L 1322:1334 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 9 30 from changes 1559:1565 arg1 position 1574:1581 the position 1570:1581 the position of the COOH-terminal cysteine 1570:1611 Moreover, changes in the position of the COOH-terminal cysteine abolished its activity. 11279095 5 31 theme insect 915:920 arg1 cells 922:926 the insect cells 911:926 the insect cells 911:926 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 7 32 theme sialylmotif 1252:1262 arg1 S 1264:1264 sialylmotif S 1252:1264 sialylmotif S 1252:1264 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 5 33 gly polysialylate 979:991 arg1 " 1021:1021 "autopolysialylation" 1001:1021 "autopolysialylation" 1001:1021 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 5 33 gly polysialylate 979:991 arg1 itself 993:998 itself 993:998 itself 993:998 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 0 34 theme bond 17:20 arg1 structures 22:31 Unique disulfide bond structures 0:31 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase 0:72 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase are required for its activity. 11279095 8 35 theme Site-directed 1406:1418 arg1 mutagenesis 1420:1430 Site-directed mutagenesis 1406:1430 Site-directed mutagenesis 1406:1430 Site-directed mutagenesis demonstrated that mutation at cysteine residues involved in the disulfide bridges completely inactivated the enzyme. 11279095 8 36 from residues 1471:1478 arg1 mutation 1450:1457 mutation 1450:1457 mutation at cysteine residues involved in the disulfide bridges 1450:1512 Site-directed mutagenesis demonstrated that mutation at cysteine residues involved in the disulfide bridges completely inactivated the enzyme. 11279095 2 37 contain contain 337:343 arg1 polysialyltransferases 270:291 two polysialyltransferases 266:291 two polysialyltransferases 266:291 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 37 contain contain 337:343 arg2 sialylmotifs 345:356 sialylmotifs L and S 345:364 sialylmotifs L and S conserved in all members of the sialyltransferases 345:415 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 37 contain contain 337:343 arg1 ST8Sia 294:299 ST8Sia II 294:302 ST8Sia II (STX) 294:308 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 37 contain contain 337:343 arg1 ST8Sia 314:319 ST8Sia IV 314:322 ST8Sia IV (PST) 314:328 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 37 contain contain 337:343 arg2 L 358:358 L 358:358 L 358:358 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 37 contain contain 337:343 arg2 S 364:364 S 364:364 S 364:364 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 1 38 theme NCAM 105:108 arg1 polysialylation 110:124 NCAM polysialylation 105:124 NCAM polysialylation 105:124 NCAM polysialylation plays a critical role in neuronal development and regeneration. 11279095 7 39 from cysteine 1240:1247 arg1 L 1230:1230 sialylmotif L 1218:1230 sialylmotif L 1218:1230 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 7 39 from cysteine 1240:1247 arg1 S 1264:1264 sialylmotif S 1252:1264 sialylmotif S 1252:1264 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 7 40 theme same 1114:1117 arg1 preparation 1126:1136 the same enzyme preparation 1110:1136 the same enzyme preparation 1110:1136 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 5 41 theme human 823:827 arg1 ST8Sia 829:834 human ST8Sia IV 823:837 human ST8Sia IV in insect cells, Trichoplusia ni, 823:871 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 7 42 theme second 1381:1386 arg1 bridge 1398:1403 a second disulfide bridge 1379:1403 a second disulfide bridge 1379:1403 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 3 43 theme sialylmotif 531:541 arg1 L 543:543 sialylmotif L 531:543 sialylmotif L 531:543 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 44 from L 543:543 arg1 S 574:574 sialylmotif S 562:574 sialylmotif S 562:574 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 7 45 theme spectrometric 1158:1170 arg1 analysis 1172:1179 mass spectrometric analysis 1153:1179 mass spectrometric analysis 1153:1179 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 3 46 from terminus 606:613 arg1 cysteine 585:592 one cysteine 581:592 one cysteine at the COOH terminus 581:613 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 46 from terminus 606:613 arg1 cysteines 518:526 three cysteines 512:526 three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus 512:613 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 46 from terminus 606:613 arg1 L 543:543 sialylmotif L 531:543 sialylmotif L 531:543 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 46 from terminus 606:613 arg1 cysteine 550:557 one cysteine 546:557 one cysteine in sialylmotif S 546:574 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 7 47 theme sialylmotif 1322:1332 arg1 L 1334:1334 sialylmotif L 1322:1334 sialylmotif L 1322:1334 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 11 48 theme close 1923:1927 arg1 proximity 1929:1937 close proximity 1923:1937 close proximity 1923:1937 These results combined indicate that the sterical structure formed by intramolecular disulfide bonds, which bring the sialylmotifs and the COOH terminus within close proximity, is critical for the catalytic activity of ST8Sia IV. 11279095 3 49 theme sialylmotif 562:572 arg1 S 574:574 sialylmotif S 562:574 sialylmotif S 562:574 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 10 50 theme fluorescence 1672:1683 arg1 protein 1685:1691 green fluorescence protein 1666:1691 green fluorescence protein 1666:1691 By contrast, the addition of green fluorescence protein at the COOH terminus of ST8Sia IV did not render the enzyme inactive. 11279095 5 51 from ST8Sia 829:834 arg1 cells 849:853 insect cells 842:853 insect cells 842:853 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 5 51 from ST8Sia 829:834 arg1 ni 869:870 Trichoplusia ni 856:870 Trichoplusia ni 856:870 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 2 52 theme molecule 234:241 arg1 Polysialylation 190:204 Polysialylation 190:204 Polysialylation of the neural cell adhesion molecule (NCAM) 190:248 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 3 53 theme gene 444:447 arg1 family 449:454 the ST8Sia gene family 433:454 the ST8Sia gene family 433:454 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 0 54 theme ST8Sia 42:47 arg1 polysialyltransferase 52:72 ST8Sia IV polysialyltransferase 42:72 ST8Sia IV polysialyltransferase 42:72 Unique disulfide bond structures found in ST8Sia IV polysialyltransferase are required for its activity. 11279095 9 55 theme COOH-terminal 1590:1602 arg1 cysteine 1604:1611 the COOH-terminal cysteine 1586:1611 the COOH-terminal cysteine 1586:1611 Moreover, changes in the position of the COOH-terminal cysteine abolished its activity. 11279095 2 56 theme cell 220:223 arg1 NCAM 244:247 NCAM 244:247 NCAM 244:247 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 56 theme cell 220:223 arg1 molecule 234:241 the neural cell adhesion molecule 209:241 the neural cell adhesion molecule (NCAM) 209:248 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 57 theme adhesion 225:232 arg1 NCAM 244:247 NCAM 244:247 NCAM 244:247 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 57 theme adhesion 225:232 arg1 molecule 234:241 the neural cell adhesion molecule 209:241 the neural cell adhesion molecule (NCAM) 209:248 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 3 58 theme family 449:454 arg1 ST8Sia 467:472 ST8Sia II 467:475 ST8Sia II 467:475 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 58 theme family 449:454 arg1 members 422:428 The members 418:428 The members 418:428 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 58 theme family 449:454 arg1 ST8Sia 481:486 ST8Sia IV 481:489 ST8Sia IV 481:489 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 3 58 theme family 449:454 arg1 unique 495:500 unique 495:500 unique 495:500 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 6 59 theme disulfide 1084:1092 arg1 bonds 1094:1098 disulfide bonds 1084:1098 disulfide bonds 1084:1098 We also found that ST8Sia IV does not form a dimer through disulfide bonds. 11279095 7 60 theme enzyme 1119:1124 arg1 preparation 1126:1136 the same enzyme preparation 1110:1136 the same enzyme preparation 1110:1136 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 9 61 theme cysteine 1604:1611 arg1 position 1574:1581 the position 1570:1581 the position of the COOH-terminal cysteine 1570:1611 Moreover, changes in the position of the COOH-terminal cysteine abolished its activity. 11279095 2 62 theme neural 213:218 arg1 NCAM 244:247 NCAM 244:247 NCAM 244:247 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 62 theme neural 213:218 arg1 molecule 234:241 the neural cell adhesion molecule 209:241 the neural cell adhesion molecule (NCAM) 209:248 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 63 dep sialylmotifs 345:356 arg1 L 358:358 L 358:358 L 358:358 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 63 dep sialylmotifs 345:356 arg1 sialylmotifs 345:356 sialylmotifs L and S 345:364 sialylmotifs L and S conserved in all members of the sialyltransferases 345:415 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 2 63 dep sialylmotifs 345:356 arg1 S 364:364 S 364:364 S 364:364 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 8 64 theme cysteine 1462:1469 arg1 residues 1471:1478 cysteine residues 1462:1478 cysteine residues involved in the disulfide bridges 1462:1512 Site-directed mutagenesis demonstrated that mutation at cysteine residues involved in the disulfide bridges completely inactivated the enzyme. 11279095 5 65 theme NCAM 938:941 arg1 polysialylation 943:957 NCAM polysialylation 938:957 NCAM polysialylation 938:957 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 7 66 theme sialylmotif 1218:1228 arg1 L 1230:1230 sialylmotif L 1218:1230 sialylmotif L 1218:1230 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 1 67 theme critical 134:141 arg1 role 143:146 a critical role 132:146 a critical role 132:146 NCAM polysialylation plays a critical role in neuronal development and regeneration. 11279095 7 68 from cysteine 1344:1351 arg1 L 1334:1334 sialylmotif L 1322:1334 sialylmotif L 1322:1334 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 10 69 theme ST8Sia 1717:1722 arg1 terminus 1705:1712 the COOH terminus 1696:1712 the COOH terminus of ST8Sia IV 1696:1725 By contrast, the addition of green fluorescence protein at the COOH terminus of ST8Sia IV did not render the enzyme inactive. 11279095 5 70 theme ST8Sia 799:804 arg1 structure/function 777:794 structure/function 777:794 structure/function 777:794 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 3 71 from cysteine 585:592 arg1 S 574:574 sialylmotif S 562:574 sialylmotif S 562:574 The members of the ST8Sia gene family, including ST8Sia II and ST8Sia IV are unique in having three cysteines in sialylmotif L, one cysteine in sialylmotif S, and one cysteine at the COOH terminus. 11279095 5 72 theme autopolysialylation 1002:1020 arg1 " 1021:1021 "autopolysialylation" 1001:1021 "autopolysialylation" 1001:1021 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 5 72 theme autopolysialylation 1002:1020 arg1 itself 993:998 itself 993:998 itself 993:998 To obtain insight into the structure/function of ST8Sia IV, we expressed human ST8Sia IV in insect cells, Trichoplusia ni, and found that the enzyme produced in the insect cells catalyzes NCAM polysialylation, although it cannot polysialylate itself ("autopolysialylation"). 11279095 7 73 theme COOH 1360:1363 arg1 terminus 1365:1372 the COOH terminus 1356:1372 the COOH terminus 1356:1372 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 11279095 10 74 theme protein 1685:1691 arg1 addition 1654:1661 the addition 1650:1661 the addition of green fluorescence protein at the COOH terminus of ST8Sia IV 1650:1725 By contrast, the addition of green fluorescence protein at the COOH terminus of ST8Sia IV did not render the enzyme inactive. 11279095 2 75 theme sialyltransferases 398:415 arg1 members 383:389 all members 379:389 all members of the sialyltransferases 379:415 Polysialylation of the neural cell adhesion molecule (NCAM) is catalyzed by two polysialyltransferases, ST8Sia II (STX) and ST8Sia IV (PST), which contain sialylmotifs L and S conserved in all members of the sialyltransferases. 11279095 7 76 theme mass 1153:1156 arg1 analysis 1172:1179 mass spectrometric analysis 1153:1179 mass spectrometric analysis 1153:1179 By using the same enzyme preparation and performing mass spectrometric analysis, we found that the first cysteine in sialylmotif L and the cysteine in sialylmotif S form a disulfide bridge, whereas the second cysteine in sialylmotif L and the cysteine at the COOH terminus form a second disulfide bridge. 20223216 6 0 theme Alanine 889:895 arg1 mutagenesis 906:916 Alanine scanning mutagenesis 889:916 Alanine scanning mutagenesis of the IL-13 interface 889:939 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 5 1 theme magnitude 806:814 arg1 increase 816:823 magnitude increase 806:823 magnitude increase 806:823 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 3 2 theme biophysical 467:477 arg1 analysis 479:486 a mutational and biophysical analysis 450:486 a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex 450:520 Here we present the crystal structure along with a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex. 20223216 1 3 theme responses 229:237 arg1 development 187:197 development 187:197 development of T helper cell type 2 (Th2) responses 187:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 0 4 theme high 90:93 arg1 complex 104:110 an unusually high affinity complex 77:110 an unusually high affinity complex between IL-13 and IL-13Ralpha2 77:141 Molecular basis for shared cytokine recognition revealed in the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2. 20223216 3 5 theme complex 514:520 arg1 analysis 479:486 a mutational and biophysical analysis 450:486 a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex 450:520 Here we present the crystal structure along with a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex. 20223216 4 6 theme binding 563:569 arg1 IL-13Ralpha1 606:617 IL-13Ralpha1 606:617 IL-13Ralpha1 606:617 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 4 6 theme binding 563:569 arg1 mode 549:552 a similar mode 539:552 a similar mode of IL-13 binding to its related signaling receptor 539:603 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 4 6 theme binding 563:569 arg1 IL-13Ralpha2 620:631 IL-13Ralpha2 620:631 IL-13Ralpha2 620:631 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 5 7 from orders 796:801 arg1 affinity 828:835 affinity 828:835 affinity 828:835 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 4 8 theme complementary 738:750 arg1 interface 752:760 a larger and more complementary interface 720:760 a larger and more complementary interface for IL-13 720:770 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 4 9 theme IL-13 557:561 arg1 binding 563:569 IL-13 binding 557:569 IL-13 binding to its related signaling receptor 557:603 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 0 10 theme complex 104:110 arg1 structure 64:72 the structure 60:72 the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2 60:141 Molecular basis for shared cytokine recognition revealed in the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2. 20223216 5 11 theme increase 816:823 arg1 orders 796:801 a four orders 789:801 a four orders of magnitude increase in affinity, to the femtomolar level 789:860 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 2 12 theme Jak/STAT 371:378 arg1 functions 390:398 canonical Jak/STAT signaling functions 361:398 canonical Jak/STAT signaling functions 361:398 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 6 13 theme prominent 1046:1054 arg1 contact 1078:1084 a prominent IL-13Ralpha2-specific contact 1044:1084 a prominent IL-13Ralpha2-specific contact 1044:1084 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 0 14 theme affinity 95:102 arg1 complex 104:110 an unusually high affinity complex 77:110 an unusually high affinity complex between IL-13 and IL-13Ralpha2 77:141 Molecular basis for shared cytokine recognition revealed in the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2. 20223216 3 15 theme IL-13/IL-13Ralpha2 495:512 arg1 complex 514:520 the IL-13/IL-13Ralpha2 complex 491:520 the IL-13/IL-13Ralpha2 complex 491:520 Here we present the crystal structure along with a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex. 20223216 1 16 theme important 173:181 arg1 Interleukin-13 144:157 Interleukin-13 144:157 Interleukin-13 144:157 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 1 16 theme important 173:181 arg1 cytokine 164:171 a cytokine 162:171 a cytokine important for development of T helper cell type 2 (Th2) responses 162:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 7 17 theme antagonists 1173:1183 arg1 development 1125:1135 development 1125:1135 development of receptor subtype-selective IL-13 antagonists 1125:1183 These results provide a framework for development of receptor subtype-selective IL-13 antagonists and indicate a decoy function for IL-13Ralpha2. 20223216 6 18 theme interface 931:939 arg1 mutagenesis 906:916 Alanine scanning mutagenesis 889:916 Alanine scanning mutagenesis of the IL-13 interface 889:939 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 4 19 theme larger 722:727 arg1 interface 752:760 a larger and more complementary interface 720:760 a larger and more complementary interface for IL-13 720:770 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 4 20 theme receptor 649:656 arg1 residues 658:665 peripheral receptor residues 638:665 peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex 638:706 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 0 21 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis for shared cytokine recognition 0:46 Molecular basis for shared cytokine recognition revealed in the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2. 20223216 7 22 theme receptor 1140:1147 arg1 antagonists 1173:1183 receptor subtype-selective IL-13 antagonists 1140:1183 receptor subtype-selective IL-13 antagonists 1140:1183 These results provide a framework for development of receptor subtype-selective IL-13 antagonists and indicate a decoy function for IL-13Ralpha2. 20223216 5 23 dep results 778:784 arg1 compared 863:870 compared 863:870 compared to IL-13Ralpha1 863:886 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 4 24 theme peripheral 638:647 arg1 residues 658:665 peripheral receptor residues 638:665 peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex 638:706 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 0 25 theme shared 20:25 arg1 recognition 36:46 shared cytokine recognition 20:46 shared cytokine recognition 20:46 Molecular basis for shared cytokine recognition revealed in the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2. 20223216 1 26 theme critical 251:258 arg1 role 260:263 a critical role 249:263 a critical role 249:263 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 3 27 theme crystal 421:427 arg1 structure 429:437 the crystal structure 417:437 the crystal structure 417:437 Here we present the crystal structure along with a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex. 20223216 7 28 theme IL-13 1167:1171 arg1 antagonists 1173:1183 receptor subtype-selective IL-13 antagonists 1140:1183 receptor subtype-selective IL-13 antagonists 1140:1183 These results provide a framework for development of receptor subtype-selective IL-13 antagonists and indicate a decoy function for IL-13Ralpha2. 20223216 4 29 theme similar 541:547 arg1 IL-13Ralpha1 606:617 IL-13Ralpha1 606:617 IL-13Ralpha1 606:617 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 4 29 theme similar 541:547 arg1 mode 549:552 a similar mode 539:552 a similar mode of IL-13 binding to its related signaling receptor 539:603 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 4 29 theme similar 541:547 arg1 IL-13Ralpha2 620:631 IL-13Ralpha2 620:631 IL-13Ralpha2 620:631 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 6 30 theme IL-13Ralpha2-specific 1056:1076 arg1 contact 1078:1084 a prominent IL-13Ralpha2-specific contact 1044:1084 a prominent IL-13Ralpha2-specific contact 1044:1084 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 1 31 theme T 202:202 arg1 responses 229:237 T helper cell type 2 (Th2) responses 202:237 T helper cell type 2 (Th2) responses 202:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 3 32 theme mutational 452:461 arg1 analysis 479:486 a mutational and biophysical analysis 450:486 a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex 450:520 Here we present the crystal structure along with a mutational and biophysical analysis of the IL-13/IL-13Ralpha2 complex. 20223216 1 33 theme helper 204:209 arg1 responses 229:237 T helper cell type 2 (Th2) responses 202:237 T helper cell type 2 (Th2) responses 202:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 5 34 theme femtomolar 845:854 arg1 level 856:860 the femtomolar level 841:860 the femtomolar level 841:860 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 2 35 theme canonical 361:369 arg1 functions 390:398 canonical Jak/STAT signaling functions 361:398 canonical Jak/STAT signaling functions 361:398 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 0 36 theme cytokine 27:34 arg1 recognition 36:46 shared cytokine recognition 20:46 shared cytokine recognition 20:46 Molecular basis for shared cytokine recognition revealed in the structure of an unusually high affinity complex between IL-13 and IL-13Ralpha2. 20223216 6 37 theme IL-13 925:929 arg1 interface 931:939 the IL-13 interface 921:939 the IL-13 interface 921:939 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 6 38 theme " 972:972 arg1 residues 974:981 several common "hotspot" residues 949:981 several common "hotspot" residues important for binding to both receptors 949:1021 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 4 39 theme IL-13/IL-13Ralpha1 681:698 arg1 complex 700:706 the IL-13/IL-13Ralpha1 complex 677:706 the IL-13/IL-13Ralpha1 complex 677:706 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 6 40 theme important 983:991 arg1 residues 974:981 several common "hotspot" residues 949:981 several common "hotspot" residues important for binding to both receptors 949:1021 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 1 41 theme cell 211:214 arg1 responses 229:237 T helper cell type 2 (Th2) responses 202:237 T helper cell type 2 (Th2) responses 202:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 7 42 theme decoy 1200:1204 arg1 function 1206:1213 a decoy function 1198:1213 a decoy function for IL-13Ralpha2 1198:1230 These results provide a framework for development of receptor subtype-selective IL-13 antagonists and indicate a decoy function for IL-13Ralpha2. 20223216 6 43 theme several 949:955 arg1 residues 974:981 several common "hotspot" residues 949:981 several common "hotspot" residues important for binding to both receptors 949:1021 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 5 44 from affinity 828:835 arg1 orders 796:801 a four orders 789:801 a four orders of magnitude increase in affinity, to the femtomolar level 789:860 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 2 45 theme Receptor 298:305 arg1 IL-13Ralpha2 315:326 IL-13Ralpha2 315:326 IL-13Ralpha2 315:326 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 2 45 theme Receptor 298:305 arg1 alpha2 307:312 The IL-13 Receptor alpha2 288:312 The IL-13 Receptor alpha2 (IL-13Ralpha2) 288:327 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 2 45 theme Receptor 298:305 arg1 receptor 334:341 a receptor 332:341 a receptor for IL-13 lacking canonical Jak/STAT signaling functions 332:398 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 7 46 theme subtype-selective 1149:1165 arg1 antagonists 1173:1183 receptor subtype-selective IL-13 antagonists 1140:1183 receptor subtype-selective IL-13 antagonists 1140:1183 These results provide a framework for development of receptor subtype-selective IL-13 antagonists and indicate a decoy function for IL-13Ralpha2. 20223216 5 47 from increase 816:823 arg1 affinity 828:835 affinity 828:835 affinity 828:835 This results in a four orders of magnitude increase in affinity, to the femtomolar level, compared to IL-13Ralpha1. 20223216 2 48 theme IL-13 292:296 arg1 IL-13Ralpha2 315:326 IL-13Ralpha2 315:326 IL-13Ralpha2 315:326 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 2 48 theme IL-13 292:296 arg1 alpha2 307:312 The IL-13 Receptor alpha2 288:312 The IL-13 Receptor alpha2 (IL-13Ralpha2) 288:327 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 2 48 theme IL-13 292:296 arg1 receptor 334:341 a receptor 332:341 a receptor for IL-13 lacking canonical Jak/STAT signaling functions 332:398 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 1 49 theme type 216:219 arg1 responses 229:237 T helper cell type 2 (Th2) responses 202:237 T helper cell type 2 (Th2) responses 202:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 20223216 2 50 theme signaling 380:388 arg1 functions 390:398 canonical Jak/STAT signaling functions 361:398 canonical Jak/STAT signaling functions 361:398 The IL-13 Receptor alpha2 (IL-13Ralpha2) is a receptor for IL-13 lacking canonical Jak/STAT signaling functions. 20223216 4 51 theme signaling 586:594 arg1 receptor 596:603 its related signaling receptor 574:603 its related signaling receptor 574:603 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 6 52 theme scanning 897:904 arg1 mutagenesis 906:916 Alanine scanning mutagenesis 889:916 Alanine scanning mutagenesis of the IL-13 interface 889:939 Alanine scanning mutagenesis of the IL-13 interface reveals several common "hotspot" residues important for binding to both receptors, but also identifies a prominent IL-13Ralpha2-specific contact. 20223216 4 53 theme related 578:584 arg1 receptor 596:603 its related signaling receptor 574:603 its related signaling receptor 574:603 While retaining a similar mode of IL-13 binding to its related signaling receptor, IL-13Ralpha1, IL-13Ralpha2 uses peripheral receptor residues unused in the IL-13/IL-13Ralpha1 complex to generate a larger and more complementary interface for IL-13. 20223216 1 54 theme Th2 224:226 arg1 responses 229:237 T helper cell type 2 (Th2) responses 202:237 T helper cell type 2 (Th2) responses 202:237 Interleukin-13 is a cytokine important for development of T helper cell type 2 (Th2) responses and plays a critical role in asthma and allergy. 22041449 6 0 theme crystal 964:970 arg1 structure 972:980 the crystal structure 960:980 the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 960:1048 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 8 1 theme NGL 1525:1527 arg1 recognition 1529:1539 NGL recognition 1525:1539 NGL recognition 1525:1539 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 7 2 theme binding 1292:1298 arg1 site 1300:1303 a Ca(2+) binding site 1283:1303 a Ca(2+) binding site in the LN domain 1283:1320 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 8 3 theme netrin 1476:1481 arg1 splicing 1497:1504 netrin G alternative splicing 1476:1504 netrin G alternative splicing 1476:1504 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 3 4 theme LN 480:481 arg1 domain 484:489 a laminin N-terminal (LN) domain 458:489 a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor 458:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 6 5 dep domain 1015:1020 arg1 LE1 1033:1035 LE1 1033:1035 LE1 1033:1035 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 6 6 theme 1.8 Å 1126:1130 arg1 resolution 1132:1141 1.8 Å resolution 1126:1141 1.8 Å resolution 1126:1141 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 3 7 theme N-terminal 468:477 arg1 domain 484:489 a laminin N-terminal (LN) domain 458:489 a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor 458:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 7 8 theme Ca 1285:1286 arg1 site 1300:1303 a Ca(2+) binding site 1283:1303 a Ca(2+) binding site in the LN domain 1283:1320 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 1 9 theme polymerization 170:183 arg1 laminins 196:203 laminins 196:203 laminins 196:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 1 9 theme polymerization 170:183 arg1 domains 185:191 the polymerization domains 166:191 the polymerization domains of laminins 166:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 7 10 theme significant 1249:1259 arg1 site 1300:1303 a Ca(2+) binding site 1283:1303 a Ca(2+) binding site in the LN domain 1283:1320 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 7 10 theme significant 1249:1259 arg1 differences 1261:1271 significant differences 1249:1271 significant differences including a Ca(2+) binding site in the LN domain 1249:1320 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 3 11 theme growth 527:532 arg1 domains 551:557 three laminin epidermal growth factor-like (LE) domains 503:557 three laminin epidermal growth factor-like (LE) domains 503:557 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 8 12 theme Gs 1395:1396 arg1 interaction 1373:1383 interaction 1373:1383 interaction of netrin Gs with NGLs 1373:1406 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 3 13 theme laminin 509:515 arg1 domains 551:557 three laminin epidermal growth factor-like (LE) domains 503:557 three laminin epidermal growth factor-like (LE) domains 503:557 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 6 14 theme anomalous 1078:1086 arg1 phasing 1100:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 7 15 theme LN 1225:1226 arg1 domains 1228:1234 laminin α chain LN domains 1209:1234 laminin α chain LN domains 1209:1234 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 4 16 contain contains 703:710 arg2 site 724:727 the binding site 712:727 the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity 712:806 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 4 16 contain contains 703:710 arg1 region 683:688 the LN domain region 669:688 the LN domain region of netrin Gs 669:701 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 6 17 theme G2 1047:1048 arg1 domain 1026:1031 domain 1026:1031 domain 1026:1031 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 6 17 theme G2 1047:1048 arg1 domain 1015:1020 the LN domain 1008:1020 the LN domain 1008:1020 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 6 17 theme G2 1047:1048 arg1 G2 1047:1048 netrin G2 1040:1048 netrin G2 1040:1048 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 1 18 theme synaptic 107:114 arg1 molecules 130:138 synaptic cell adhesion molecules 107:138 synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins 107:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 7 19 theme α 1217:1217 arg1 chain 1219:1223 laminin α chain 1209:1223 laminin α chain LN domains 1209:1234 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 0 20 theme netrin 50:55 arg1 G2 57:58 netrin G2 50:58 netrin G2 50:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 2 21 theme G 217:217 arg1 proteins 219:226 Two netrin G proteins 206:226 Two netrin G proteins 206:226 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 1 22 theme adhesion 121:128 arg1 molecules 130:138 synaptic cell adhesion molecules 107:138 synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins 107:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 1 23 theme related 140:146 arg1 molecules 130:138 synaptic cell adhesion molecules 107:138 synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins 107:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 8 24 theme binding 1356:1362 arg1 unit 1364:1367 the minimal binding unit 1344:1367 the minimal binding unit for interaction of netrin Gs with NGLs 1344:1406 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 4 25 theme domain 676:681 arg1 region 683:688 the LN domain region 669:688 the LN domain region of netrin Gs 669:701 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 2 26 theme G 346:346 arg1 NGL 356:358 netrin G ligand (NGL)-1 339:361 netrin G ligand (NGL)-1 339:361 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 26 theme G 346:346 arg1 proteins 330:337 the leucine-rich repeat proteins 306:337 the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2 306:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 1 27 theme Netrin 61:66 arg1 proteins 70:77 Netrin G proteins 61:77 Netrin G proteins 61:77 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 7 28 theme laminin 1209:1215 arg1 chain 1219:1223 laminin α chain 1209:1223 laminin α chain LN domains 1209:1234 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 0 29 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the ligand binding domain of netrin G2 0:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 6 30 theme sulfur 1053:1058 arg1 phasing 1100:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 3 31 theme Netrin 388:393 arg1 proteins 397:404 Netrin G proteins 388:404 Netrin G proteins 388:404 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 3 32 theme laminin 460:466 arg1 domain 484:489 a laminin N-terminal (LN) domain 458:489 a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor 458:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 5 33 theme domain 857:862 arg1 regions 864:870 their LE domain regions 848:870 their LE domain regions 848:870 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 7 34 theme similar 1190:1196 arg1 architecture 1177:1188 an overall architecture 1166:1188 an overall architecture similar to that of laminin α chain LN domains 1166:1234 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 3 35 theme C' 565:566 arg1 region 568:573 a C' region 563:573 a C' region containing a glycosylphosphatidylinositol anchor 563:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 8 36 with interaction 1373:1383 arg1 NGLs 1403:1406 NGLs 1403:1406 NGLs 1403:1406 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 0 37 theme ligand 25:30 arg1 domain 40:45 the ligand binding domain 21:45 the ligand binding domain of netrin G2 21:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 0 37 theme ligand 25:30 arg1 G2 57:58 netrin G2 50:58 netrin G2 50:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 7 38 theme overall 1169:1175 arg1 architecture 1177:1188 an overall architecture 1166:1188 an overall architecture similar to that of laminin α chain LN domains 1166:1234 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 0 39 theme domain 40:45 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the ligand binding domain of netrin G2 0:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 3 40 theme glycosylphosphatidylinositol 588:615 arg1 anchor 617:622 a glycosylphosphatidylinositol anchor 586:622 a glycosylphosphatidylinositol anchor 586:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 2 41 dep proteins 330:337 arg1 NGL-2 367:371 NGL-2 367:371 NGL-2 367:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 41 dep proteins 330:337 arg1 NGL 356:358 netrin G ligand (NGL)-1 339:361 netrin G ligand (NGL)-1 339:361 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 41 dep proteins 330:337 arg1 proteins 330:337 the leucine-rich repeat proteins 306:337 the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2 306:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 8 42 theme G 1483:1483 arg1 splicing 1497:1504 netrin G alternative splicing 1476:1504 netrin G alternative splicing 1476:1504 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 1 43 theme laminins 196:203 arg1 laminins 196:203 laminins 196:203 laminins 196:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 1 43 theme laminins 196:203 arg1 domains 185:191 the polymerization domains 166:191 the polymerization domains of laminins 166:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 2 44 theme leucine-rich 310:321 arg1 NGL-2 367:371 NGL-2 367:371 NGL-2 367:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 44 theme leucine-rich 310:321 arg1 NGL 356:358 netrin G ligand (NGL)-1 339:361 netrin G ligand (NGL)-1 339:361 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 44 theme leucine-rich 310:321 arg1 proteins 330:337 the leucine-rich repeat proteins 306:337 the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2 306:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 6 45 theme LN 1012:1013 arg1 domain 1015:1020 the LN domain 1008:1020 the LN domain 1008:1020 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 6 45 theme LN 1012:1013 arg1 G2 1047:1048 netrin G2 1040:1048 netrin G2 1040:1048 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 3 46 theme multi-domain 421:432 arg1 architecture 434:445 a common multi-domain architecture 412:445 a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor 412:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 7 47 theme LN 1312:1313 arg1 domain 1315:1320 the LN domain 1308:1320 the LN domain 1308:1320 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 6 48 theme netrin 1040:1045 arg1 G2 1047:1048 netrin G2 1040:1048 netrin G2 1040:1048 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 2 49 theme vertebrate 243:252 arg1 genomes 254:260 vertebrate genomes 243:260 vertebrate genomes 243:260 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 49 theme vertebrate 243:252 arg1 netrins 263:269 netrins G1 and G2 263:279 netrins G1 and G2 263:279 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 5 50 theme Netrin 809:814 arg1 Gs 816:817 Netrin Gs 809:817 Netrin Gs 809:817 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 3 51 theme epidermal 517:525 arg1 domains 551:557 three laminin epidermal growth factor-like (LE) domains 503:557 three laminin epidermal growth factor-like (LE) domains 503:557 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 1 52 theme small 91:95 arg1 family 97:102 a small family 89:102 a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins 89:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 4 53 theme sub-micromolar 784:797 arg1 affinity 799:806 sub-micromolar affinity 784:806 sub-micromolar affinity 784:806 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 6 54 theme diffraction 1088:1098 arg1 phasing 1100:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 2 55 dep netrins 263:269 arg1 G1 271:272 G1 271:272 G1 271:272 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 55 dep netrins 263:269 arg1 netrins 263:269 netrins G1 and G2 263:279 netrins G1 and G2 263:279 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 55 dep netrins 263:269 arg1 G2 278:279 G2 278:279 G2 278:279 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 4 56 theme Gs 700:701 arg1 region 683:688 the LN domain region 669:688 the LN domain region of netrin Gs 669:701 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 3 57 theme LE 547:548 arg1 domains 551:557 three laminin epidermal growth factor-like (LE) domains 503:557 three laminin epidermal growth factor-like (LE) domains 503:557 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 6 58 theme single-wavelength 1060:1076 arg1 phasing 1100:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 sulfur single-wavelength anomalous diffraction phasing 1053:1106 We determined the crystal structure for a fragment comprising the LN domain and domain LE1 of netrin G2 by sulfur single-wavelength anomalous diffraction phasing and refined it to 1.8 Å resolution. 22041449 4 59 theme deletion 638:645 arg1 analysis 647:654 deletion analysis 638:654 deletion analysis 638:654 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 0 60 theme G2 57:58 arg1 domain 40:45 the ligand binding domain 21:45 the ligand binding domain of netrin G2 21:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 0 60 theme G2 57:58 arg1 G2 57:58 netrin G2 50:58 netrin G2 50:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 5 61 theme splice 932:937 arg1 forms 939:943 all splice forms 928:943 all splice forms 928:943 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 3 62 theme factor-like 534:544 arg1 domains 551:557 three laminin epidermal growth factor-like (LE) domains 503:557 three laminin epidermal growth factor-like (LE) domains 503:557 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 1 63 theme cell 116:119 arg1 molecules 130:138 synaptic cell adhesion molecules 107:138 synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins 107:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 8 64 theme minimal 1348:1354 arg1 unit 1364:1367 the minimal binding unit 1344:1367 the minimal binding unit for interaction of netrin Gs with NGLs 1344:1406 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 7 65 theme chain 1219:1223 arg1 domains 1228:1234 laminin α chain LN domains 1209:1234 laminin α chain LN domains 1209:1234 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 5 66 from identical 915:923 arg1 forms 939:943 all splice forms 928:943 all splice forms 928:943 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 1 67 theme molecules 130:138 arg1 family 97:102 a small family 89:102 a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins 89:203 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 7 68 from site 1300:1303 arg1 domain 1315:1320 the LN domain 1308:1320 the LN domain 1308:1320 The structure reveals an overall architecture similar to that of laminin α chain LN domains but includes significant differences including a Ca(2+) binding site in the LN domain. 22041449 4 69 theme LN 673:674 arg1 region 683:688 the LN domain region 669:688 the LN domain region of netrin Gs 669:701 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 5 70 from forms 939:943 arg1 identical 915:923 identical 915:923 identical 915:923 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 5 70 from forms 939:943 arg1 region 889:894 the binding region 877:894 the binding region 877:894 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 8 71 theme netrin 1388:1393 arg1 Gs 1395:1396 netrin Gs 1388:1396 netrin Gs 1388:1396 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 2 72 theme netrin 339:344 arg1 NGL 356:358 netrin G ligand (NGL)-1 339:361 netrin G ligand (NGL)-1 339:361 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 72 theme netrin 339:344 arg1 proteins 330:337 the leucine-rich repeat proteins 306:337 the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2 306:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 1 73 theme G 68:68 arg1 proteins 70:77 Netrin G proteins 61:77 Netrin G proteins 61:77 Netrin G proteins represent a small family of synaptic cell adhesion molecules related to netrins and to the polymerization domains of laminins. 22041449 2 74 theme repeat 323:328 arg1 NGL-2 367:371 NGL-2 367:371 NGL-2 367:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 74 theme repeat 323:328 arg1 NGL 356:358 netrin G ligand (NGL)-1 339:361 netrin G ligand (NGL)-1 339:361 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 74 theme repeat 323:328 arg1 proteins 330:337 the leucine-rich repeat proteins 306:337 the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2 306:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 5 75 theme LE 854:855 arg1 regions 864:870 their LE domain regions 848:870 their LE domain regions 848:870 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 2 76 theme netrin 210:215 arg1 proteins 219:226 Two netrin G proteins 206:226 Two netrin G proteins 206:226 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 3 77 theme G 395:395 arg1 proteins 397:404 Netrin G proteins 388:404 Netrin G proteins 388:404 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 0 78 theme binding 32:38 arg1 domain 40:45 the ligand binding domain 21:45 the ligand binding domain of netrin G2 21:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 0 78 theme binding 32:38 arg1 G2 57:58 netrin G2 50:58 netrin G2 50:58 Crystal structure of the ligand binding domain of netrin G2. 22041449 5 79 theme LN 901:902 arg1 domain 904:909 the LN domain 897:909 the LN domain 897:909 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 5 79 theme LN 901:902 arg1 region 889:894 the binding region 877:894 the binding region 877:894 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 8 80 theme structural 1416:1425 arg1 features 1427:1434 structural features 1416:1434 structural features specific to netrin Gs 1416:1456 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 4 81 theme binding 716:722 arg1 site 724:727 the binding site 712:727 the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity 712:806 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 22041449 2 82 theme ligand 348:353 arg1 NGL 356:358 netrin G ligand (NGL)-1 339:361 netrin G ligand (NGL)-1 339:361 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 2 82 theme ligand 348:353 arg1 proteins 330:337 the leucine-rich repeat proteins 306:337 the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2 306:371 Two netrin G proteins are encoded in vertebrate genomes, netrins G1 and G2, which are known to bind the leucine-rich repeat proteins netrin G ligand (NGL)-1 and NGL-2, respectively. 22041449 8 83 theme specific 1436:1443 arg1 features 1427:1434 structural features 1416:1434 structural features specific to netrin Gs 1416:1456 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 3 84 theme common 414:419 arg1 architecture 434:445 a common multi-domain architecture 412:445 a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor 412:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 8 85 theme alternative 1485:1495 arg1 splicing 1497:1504 netrin G alternative splicing 1476:1504 netrin G alternative splicing 1476:1504 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 3 86 contain containing 575:584 arg2 anchor 617:622 a glycosylphosphatidylinositol anchor 586:622 a glycosylphosphatidylinositol anchor 586:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 3 86 contain containing 575:584 arg1 region 568:573 a C' region 563:573 a C' region containing a glycosylphosphatidylinositol anchor 563:622 Netrin G proteins share a common multi-domain architecture comprising a laminin N-terminal (LN) domain followed by three laminin epidermal growth factor-like (LE) domains and a C' region containing a glycosylphosphatidylinositol anchor. 22041449 5 87 theme binding 881:887 arg1 domain 904:909 the LN domain 897:909 the LN domain 897:909 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 5 87 theme binding 881:887 arg1 identical 915:923 identical 915:923 identical 915:923 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 5 87 theme binding 881:887 arg1 region 889:894 the binding region 877:894 the binding region 877:894 Netrin Gs are alternatively spliced in their LE domain regions, but the binding region, the LN domain, is identical in all splice forms. 22041449 8 88 theme netrin 1448:1453 arg1 Gs 1455:1456 netrin Gs 1448:1456 netrin Gs 1448:1456 These results reveal the minimal binding unit for interaction of netrin Gs with NGLs, define structural features specific to netrin Gs, and suggest that netrin G alternative splicing is not involved in NGL recognition. 22041449 4 89 theme netrin 693:698 arg1 Gs 700:701 netrin Gs 693:701 netrin Gs 693:701 Here, we use deletion analysis to show that the LN domain region of netrin Gs contains the binding site for NGLs to which they bind with 1:1 stoichiometry and sub-micromolar affinity. 30140003 7 0 gly N-glycoprotein 875:888 arg1 N-glycoprotein 875:888 the substrate N-glycoprotein 861:888 the substrate N-glycoprotein 861:888 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 30140003 6 1 theme aromatic 674:681 arg1 rings 683:687 two aromatic rings 670:687 two aromatic rings 670:687 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 8 2 theme GnT-V 1108:1112 arg1 inhibition 1094:1103 the previously observed inhibition 1070:1103 the previously observed inhibition of GnT-V by branching enzyme GnT-III 1070:1140 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 5 3 theme accessary 636:644 arg1 domains 646:652 two accessary domains 632:652 two accessary domains 632:652 GnT-V luminal domain is composed of a GT-B fold and two accessary domains. 30140003 1 4 theme β1-6-linked 205:215 arg1 branch 237:242 a β1-6-linked N-acetylglucosamine branch 203:242 a β1-6-linked N-acetylglucosamine branch 203:242 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 7 5 theme protein-selective 923:939 arg1 modification 966:977 protein-selective and site-specific glycan modification 923:977 protein-selective and site-specific glycan modification 923:977 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 30140003 4 6 theme crystal 506:512 arg1 structures 514:523 crystal structures 506:523 crystal structures of human GnT-V luminal domain with a substrate analog 506:577 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 2 7 from formation 257:265 arg1 receptors 283:291 cell surface receptors 270:291 cell surface receptors 270:291 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 4 8 theme substrate 562:570 arg1 analog 572:577 a substrate analog 560:577 a substrate analog 560:577 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 8 9 theme enzyme 1127:1132 arg1 GnT-III 1134:1140 branching enzyme GnT-III 1117:1140 branching enzyme GnT-III 1117:1140 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 2 10 theme drug 360:363 arg1 development 365:375 drug development 360:375 drug development 360:375 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 3 11 theme catalytic 418:426 arg1 mechanism 428:436 GnT-V's catalytic mechanism 410:436 GnT-V's catalytic mechanism 410:436 However, the molecular basis of GnT-V's catalytic mechanism and substrate specificity are not fully understood. 30140003 4 12 theme GnT-V 534:538 arg1 domain 548:553 human GnT-V luminal domain 528:553 human GnT-V luminal domain 528:553 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 8 13 theme branching 1117:1125 arg1 GnT-III 1134:1140 branching enzyme GnT-III 1117:1140 branching enzyme GnT-III 1117:1140 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 6 14 theme global 756:761 arg1 conformation 763:774 the global conformation 752:774 the global conformation 752:774 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 4 15 theme human 528:532 arg1 GnT-V 534:538 human GnT-V 528:538 human GnT-V luminal domain 528:553 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 7 16 theme glycan 959:964 arg1 modification 966:977 protein-selective and site-specific glycan modification 923:977 protein-selective and site-specific glycan modification 923:977 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 30140003 2 17 theme promising 339:347 arg1 target 349:354 a promising target 337:354 a promising target for drug development 337:375 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 1 18 theme N-acetylglucosamine 217:235 arg1 branch 237:242 a β1-6-linked N-acetylglucosamine branch 203:242 a β1-6-linked N-acetylglucosamine branch 203:242 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 4 19 theme domain 548:553 arg1 structures 514:523 crystal structures 506:523 crystal structures of human GnT-V luminal domain with a substrate analog 506:577 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 5 20 theme GT-B 618:621 arg1 fold 623:626 a GT-B fold 616:626 a GT-B fold 616:626 GnT-V luminal domain is composed of a GT-B fold and two accessary domains. 30140003 4 21 with structures 514:523 arg1 analog 572:577 a substrate analog 560:577 a substrate analog 560:577 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 3 22 theme substrate 442:450 arg1 specificity 452:462 substrate specificity 442:462 substrate specificity 442:462 However, the molecular basis of GnT-V's catalytic mechanism and substrate specificity are not fully understood. 30140003 4 23 theme luminal 540:546 arg1 domain 548:553 human GnT-V luminal domain 528:553 human GnT-V luminal domain 528:553 Here, we report crystal structures of human GnT-V luminal domain with a substrate analog. 30140003 8 24 theme N-glycan 1207:1214 arg1 branching 1216:1224 N-glycan branching 1207:1224 N-glycan branching 1207:1224 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 8 25 theme rational 1172:1179 arg1 design 1181:1186 the rational design 1168:1186 the rational design of drugs targeting N-glycan branching 1168:1224 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 6 26 theme GnT-V 826:830 arg1 specificity 811:821 the fine branch specificity 795:821 the fine branch specificity of GnT-V 795:830 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 6 27 theme N-glycan 730:737 arg1 branch 707:712 the α1-6 branch 698:712 the α1-6 branch of the acceptor N-glycan 698:737 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 7 28 with interaction 846:856 arg1 GnT-V 895:899 GnT-V 895:899 GnT-V 895:899 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 30140003 6 29 theme acceptor 721:728 arg1 N-glycan 730:737 the acceptor N-glycan 717:737 the acceptor N-glycan 717:737 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 1 30 link β1-6-linked 205:215 arg1 branch 237:242 a β1-6-linked N-acetylglucosamine branch 203:242 a β1-6-linked N-acetylglucosamine branch 203:242 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 2 31 theme β1-6 245:248 arg1 formation 257:265 β1-6 branch formation 245:265 β1-6 branch formation on cell surface receptors 245:291 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 1 32 theme specific 146:153 arg1 N-glycans 155:163 specific N-glycans 146:163 specific N-glycans 146:163 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 7 33 theme N-glycoprotein 875:888 arg1 interaction 846:856 interaction 846:856 interaction of the substrate N-glycoprotein with GnT-V 846:899 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 30140003 0 34 theme N-acetylglucosaminyltransferase-V 45:77 arg1 mechanism 14:22 mechanism 14:22 mechanism 14:22 Structure and mechanism of cancer-associated N-acetylglucosaminyltransferase-V. 30140003 0 34 theme N-acetylglucosaminyltransferase-V 45:77 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and mechanism of cancer-associated N-acetylglucosaminyltransferase-V. 30140003 2 35 theme cancer 305:310 arg1 metastasis 312:321 cancer metastasis 305:321 cancer metastasis 305:321 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 7 36 theme substrate 865:873 arg1 N-glycoprotein 875:888 the substrate N-glycoprotein 861:888 the substrate N-glycoprotein 861:888 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 30140003 0 37 theme cancer-associated 27:43 arg1 N-acetylglucosaminyltransferase-V 45:77 cancer-associated N-acetylglucosaminyltransferase-V 27:77 cancer-associated N-acetylglucosaminyltransferase-V 27:77 Structure and mechanism of cancer-associated N-acetylglucosaminyltransferase-V. 30140003 8 38 theme catalytic 1040:1048 arg1 mechanism 1050:1058 a catalytic mechanism 1038:1058 a catalytic mechanism 1038:1058 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 6 39 theme branch 804:809 arg1 specificity 811:821 the fine branch specificity 795:821 the fine branch specificity of GnT-V 795:830 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 1 40 theme N-glycans 155:163 arg1 structure 133:141 the structure 129:141 the structure of specific N-glycans 129:163 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 1 41 link α1-6-linked 178:188 arg1 mannose 190:196 α1-6-linked mannose 178:196 α1-6-linked mannose 178:196 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 2 42 theme cell 270:273 arg1 receptors 283:291 cell surface receptors 270:291 cell surface receptors 270:291 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 2 43 theme surface 275:281 arg1 receptors 283:291 cell surface receptors 270:291 cell surface receptors 270:291 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 6 44 theme fine 799:802 arg1 specificity 811:821 the fine branch specificity 795:821 the fine branch specificity of GnT-V 795:830 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 8 45 theme drugs 1191:1195 arg1 design 1181:1186 the rational design 1168:1186 the rational design of drugs targeting N-glycan branching 1168:1224 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 8 46 theme observed 1085:1092 arg1 inhibition 1094:1103 the previously observed inhibition 1070:1103 the previously observed inhibition of GnT-V by branching enzyme GnT-III 1070:1140 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 8 47 theme acceptor-GnT-V 996:1009 arg1 structure 1019:1027 the acceptor-GnT-V complex structure 992:1027 the acceptor-GnT-V complex structure 992:1027 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 6 48 theme α1-6 702:705 arg1 branch 707:712 the α1-6 branch 698:712 the α1-6 branch of the acceptor N-glycan 698:737 Interestingly, two aromatic rings sandwich the α1-6 branch of the acceptor N-glycan and restrain the global conformation, partly explaining the fine branch specificity of GnT-V. 30140003 1 49 theme α1-6-linked 178:188 arg1 mannose 190:196 α1-6-linked mannose 178:196 α1-6-linked mannose 178:196 N-acetylglucosaminyltransferase-V (GnT-V) alters the structure of specific N-glycans by modifying α1-6-linked mannose with a β1-6-linked N-acetylglucosamine branch. 30140003 3 50 theme molecular 391:399 arg1 basis 401:405 the molecular basis 387:405 the molecular basis of GnT-V's catalytic mechanism and substrate specificity 387:462 However, the molecular basis of GnT-V's catalytic mechanism and substrate specificity are not fully understood. 30140003 2 51 theme branch 250:255 arg1 formation 257:265 β1-6 branch formation 245:265 β1-6 branch formation on cell surface receptors 245:291 β1-6 branch formation on cell surface receptors accelerates cancer metastasis, making GnT-V a promising target for drug development. 30140003 5 52 theme GnT-V 580:584 arg1 domain 594:599 GnT-V luminal domain 580:599 GnT-V luminal domain 580:599 GnT-V luminal domain is composed of a GT-B fold and two accessary domains. 30140003 5 53 theme luminal 586:592 arg1 domain 594:599 GnT-V luminal domain 580:599 GnT-V luminal domain 580:599 GnT-V luminal domain is composed of a GT-B fold and two accessary domains. 30140003 8 54 theme complex 1011:1017 arg1 structure 1019:1027 the acceptor-GnT-V complex structure 992:1027 the acceptor-GnT-V complex structure 992:1027 In summary, the acceptor-GnT-V complex structure suggests a catalytic mechanism, explains the previously observed inhibition of GnT-V by branching enzyme GnT-III, and provides a basis for the rational design of drugs targeting N-glycan branching. 30140003 7 55 theme site-specific 945:957 arg1 modification 966:977 protein-selective and site-specific glycan modification 923:977 protein-selective and site-specific glycan modification 923:977 In addition, interaction of the substrate N-glycoprotein with GnT-V likely contributes to protein-selective and site-specific glycan modification. 29374258 4 0 theme domain 736:741 arg1 insertion 743:751 a tandem transmembrane domain insertion 713:751 a tandem transmembrane domain insertion into its glycosyltransferase-A fold 713:787 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 1 1 theme cell 168:171 arg1 surface 173:179 the cell surface 164:179 the cell surface 164:179 Many eukaryotic proteins are anchored to the cell surface via the glycolipid glycosylphosphatidylinositol (GPI). 29374258 5 2 theme Mutational 827:836 arg1 analysis 838:845 Mutational analysis 827:845 Mutational analysis 827:845 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 7 3 theme PGAP4 1192:1196 arg1 structure 1179:1187 the structure 1175:1187 the structure of PGAP4 1175:1196 In summary, we present insights into the structure of PGAP4 and elucidate the initial step of GPI-GalNAc biosynthesis. 29374258 4 4 theme transmembrane 722:734 arg1 insertion 743:751 a tandem transmembrane domain insertion 713:751 a tandem transmembrane domain insertion into its glycosyltransferase-A fold 713:787 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 5 theme Golgi 595:599 arg1 glycosyltransferases 601:620 known Golgi glycosyltransferases 589:620 known Golgi glycosyltransferases 589:620 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 2 6 theme GalNAc 316:321 arg1 modifications 324:336 diverse N-acetylgalactosamine (GalNAc) modifications 285:336 diverse N-acetylgalactosamine (GalNAc) modifications 285:336 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 3 7 theme Golgi-resident 406:419 arg1 PGAP4 444:448 the Golgi-resident GPI-GalNAc transferase PGAP4 402:448 the Golgi-resident GPI-GalNAc transferase PGAP4 402:448 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 0 8 theme catalytic 105:113 arg1 domain 115:120 the catalytic domain 101:120 the catalytic domain 101:120 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 4 9 theme comparative 805:815 arg1 modeling 817:824 comparative modeling 805:824 comparative modeling 805:824 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 2 10 theme unresolved 365:374 arg1 process 376:382 a yet unresolved process 359:382 a yet unresolved process 359:382 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 6 11 theme various 1044:1050 arg1 proteins 1065:1072 various GPI-anchored proteins 1044:1072 various GPI-anchored proteins 1044:1072 We suggest that a juxtamembrane region of PGAP4 accommodates various GPI-anchored proteins, presenting their acceptor residue toward the catalytic center. 29374258 3 12 theme GPI-GalNAc 511:520 arg1 structures 522:531 GPI-GalNAc structures 511:531 GPI-GalNAc structures 511:531 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 6 13 theme catalytic 1120:1128 arg1 center 1130:1135 the catalytic center 1116:1135 the catalytic center 1116:1135 We suggest that a juxtamembrane region of PGAP4 accommodates various GPI-anchored proteins, presenting their acceptor residue toward the catalytic center. 29374258 2 14 theme N-acetylgalactosamine 293:313 arg1 modifications 324:336 diverse N-acetylgalactosamine (GalNAc) modifications 285:336 diverse N-acetylgalactosamine (GalNAc) modifications 285:336 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 5 15 theme catalytic 857:865 arg1 site 867:870 a catalytic site 855:870 a catalytic site 855:870 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 3 16 theme transferase 432:442 arg1 PGAP4 444:448 the Golgi-resident GPI-GalNAc transferase PGAP4 402:448 the Golgi-resident GPI-GalNAc transferase PGAP4 402:448 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 2 17 theme diverse 285:291 arg1 modifications 324:336 diverse N-acetylgalactosamine (GalNAc) modifications 285:336 diverse N-acetylgalactosamine (GalNAc) modifications 285:336 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 3 18 theme mass 462:465 arg1 spectrometry 467:478 mass spectrometry 462:478 mass spectrometry 462:478 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 7 19 theme GPI-GalNAc 1232:1241 arg1 biosynthesis 1243:1254 GPI-GalNAc biosynthesis 1232:1254 GPI-GalNAc biosynthesis 1232:1254 In summary, we present insights into the structure of PGAP4 and elucidate the initial step of GPI-GalNAc biosynthesis. 29374258 1 20 theme glycolipid 189:198 arg1 GPI 230:232 GPI 230:232 GPI 230:232 Many eukaryotic proteins are anchored to the cell surface via the glycolipid glycosylphosphatidylinositol (GPI). 29374258 1 20 theme glycolipid 189:198 arg1 glycosylphosphatidylinositol 200:227 the glycolipid glycosylphosphatidylinositol 185:227 the glycolipid glycosylphosphatidylinositol (GPI) 185:233 Many eukaryotic proteins are anchored to the cell surface via the glycolipid glycosylphosphatidylinositol (GPI). 29374258 7 21 theme biosynthesis 1243:1254 arg1 step 1224:1227 the initial step 1212:1227 the initial step of GPI-GalNAc biosynthesis 1212:1254 In summary, we present insights into the structure of PGAP4 and elucidate the initial step of GPI-GalNAc biosynthesis. 29374258 5 22 theme several 924:930 arg1 residues 932:939 several residues 924:939 several residues potentially involved in acceptor binding 924:980 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 0 23 theme Golgi 20:24 arg1 transferase 52:62 a Golgi GPI-N-acetylgalactosamine transferase 18:62 a Golgi GPI-N-acetylgalactosamine transferase 18:62 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 6 24 theme PGAP4 1025:1029 arg1 region 1015:1020 a juxtamembrane region 999:1020 a juxtamembrane region of PGAP4 999:1029 We suggest that a juxtamembrane region of PGAP4 accommodates various GPI-anchored proteins, presenting their acceptor residue toward the catalytic center. 29374258 3 25 theme PGAP4 485:489 arg1 cells 500:504 PGAP4 knockout cells 485:504 PGAP4 knockout cells 485:504 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 5 26 theme DXD-like 875:882 arg1 motif 884:888 a DXD-like motif 873:888 a DXD-like motif for UDP-GalNAc donor binding 873:917 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 4 27 theme known 589:593 arg1 glycosyltransferases 601:620 known Golgi glycosyltransferases 589:620 known Golgi glycosyltransferases 589:620 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 28 contain contains 665:672 arg1 PGAP4 567:571 PGAP4 567:571 PGAP4 567:571 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 28 contain contains 665:672 arg2 domains 694:700 three transmembrane domains 674:700 three transmembrane domains 674:700 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 28 contain contains 665:672 arg1 protein 653:659 a single-pass membrane protein 630:659 a single-pass membrane protein 630:659 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 28 contain contains 665:672 arg2 insertion 743:751 a tandem transmembrane domain insertion 713:751 a tandem transmembrane domain insertion into its glycosyltransferase-A fold 713:787 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 1 29 theme Many 123:126 arg1 proteins 139:146 Many eukaryotic proteins 123:146 Many eukaryotic proteins 123:146 Many eukaryotic proteins are anchored to the cell surface via the glycolipid glycosylphosphatidylinositol (GPI). 29374258 3 30 theme knockout 491:498 arg1 cells 500:504 PGAP4 knockout cells 485:504 PGAP4 knockout cells 485:504 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 0 31 theme transferase 52:62 arg1 Identification 0:13 Identification 0:13 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 0:121 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 5 32 theme UDP-GalNAc 894:903 arg1 binding 911:917 UDP-GalNAc donor binding 894:917 UDP-GalNAc donor binding 894:917 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 0 33 theme GPI-N-acetylgalactosamine 26:50 arg1 transferase 52:62 a Golgi GPI-N-acetylgalactosamine transferase 18:62 a Golgi GPI-N-acetylgalactosamine transferase 18:62 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 6 34 theme juxtamembrane 1001:1013 arg1 region 1015:1020 a juxtamembrane region 999:1020 a juxtamembrane region of PGAP4 999:1029 We suggest that a juxtamembrane region of PGAP4 accommodates various GPI-anchored proteins, presenting their acceptor residue toward the catalytic center. 29374258 7 35 theme initial 1216:1222 arg1 step 1224:1227 the initial step 1212:1227 the initial step of GPI-GalNAc biosynthesis 1212:1254 In summary, we present insights into the structure of PGAP4 and elucidate the initial step of GPI-GalNAc biosynthesis. 29374258 0 36 from regions 90:96 arg1 domain 115:120 the catalytic domain 101:120 the catalytic domain 101:120 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 4 37 theme membrane 644:651 arg1 PGAP4 567:571 PGAP4 567:571 PGAP4 567:571 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 37 theme membrane 644:651 arg1 protein 653:659 a single-pass membrane protein 630:659 a single-pass membrane protein 630:659 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 5 38 theme donor 905:909 arg1 binding 911:917 UDP-GalNAc donor binding 894:917 UDP-GalNAc donor binding 894:917 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 0 39 theme tandem 69:74 arg1 regions 90:96 tandem transmembrane regions 69:96 tandem transmembrane regions in the catalytic domain 69:120 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 1 40 theme eukaryotic 128:137 arg1 proteins 139:146 Many eukaryotic proteins 123:146 Many eukaryotic proteins 123:146 Many eukaryotic proteins are anchored to the cell surface via the glycolipid glycosylphosphatidylinositol (GPI). 29374258 3 41 theme GPI-GalNAc 421:430 arg1 PGAP4 444:448 the Golgi-resident GPI-GalNAc transferase PGAP4 402:448 the Golgi-resident GPI-GalNAc transferase PGAP4 402:448 Here we identify the Golgi-resident GPI-GalNAc transferase PGAP4 and show by mass spectrometry that PGAP4 knockout cells lose GPI-GalNAc structures. 29374258 2 42 theme Mammalian 236:244 arg1 GPIs 246:249 Mammalian GPIs 236:249 Mammalian GPIs 236:249 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 4 43 dep glycosyltransferases 601:620 arg1 contrast 577:584 contrast 577:584 contrast 577:584 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 0 44 with Identification 0:13 arg1 regions 90:96 tandem transmembrane regions 69:96 tandem transmembrane regions in the catalytic domain 69:120 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 4 45 theme glycosyltransferase-A 762:782 arg1 fold 784:787 its glycosyltransferase-A fold 758:787 its glycosyltransferase-A fold 758:787 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 6 46 theme GPI-anchored 1052:1063 arg1 proteins 1065:1072 various GPI-anchored proteins 1044:1072 various GPI-anchored proteins 1044:1072 We suggest that a juxtamembrane region of PGAP4 accommodates various GPI-anchored proteins, presenting their acceptor residue toward the catalytic center. 29374258 5 47 theme acceptor 965:972 arg1 binding 974:980 acceptor binding 965:980 acceptor binding 965:980 Mutational analysis reveals a catalytic site, a DXD-like motif for UDP-GalNAc donor binding, and several residues potentially involved in acceptor binding. 29374258 7 48 attach present 1153:1159 arg2 we 1150:1151 we 1150:1151 we 1150:1151 In summary, we present insights into the structure of PGAP4 and elucidate the initial step of GPI-GalNAc biosynthesis. 29374258 7 48 attach present 1153:1159 arg1 summary 1141:1147 summary 1141:1147 summary 1141:1147 In summary, we present insights into the structure of PGAP4 and elucidate the initial step of GPI-GalNAc biosynthesis. 29374258 6 49 theme acceptor 1092:1099 arg1 residue 1101:1107 their acceptor residue 1086:1107 their acceptor residue 1086:1107 We suggest that a juxtamembrane region of PGAP4 accommodates various GPI-anchored proteins, presenting their acceptor residue toward the catalytic center. 29374258 4 50 theme tandem 715:720 arg1 insertion 743:751 a tandem transmembrane domain insertion 713:751 a tandem transmembrane domain insertion into its glycosyltransferase-A fold 713:787 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 51 theme transmembrane 680:692 arg1 domains 694:700 three transmembrane domains 674:700 three transmembrane domains 674:700 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 51 theme transmembrane 680:692 arg1 insertion 743:751 a tandem transmembrane domain insertion 713:751 a tandem transmembrane domain insertion into its glycosyltransferase-A fold 713:787 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 2 52 contain have 251:254 arg2 core 268:271 a conserved core 256:271 a conserved core 256:271 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 2 52 contain have 251:254 arg1 GPIs 246:249 Mammalian GPIs 236:249 Mammalian GPIs 236:249 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 0 53 theme transmembrane 76:88 arg1 regions 90:96 tandem transmembrane regions 69:96 tandem transmembrane regions in the catalytic domain 69:120 Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain. 29374258 2 54 theme conserved 258:266 arg1 core 268:271 a conserved core 256:271 a conserved core 256:271 Mammalian GPIs have a conserved core but exhibit diverse N-acetylgalactosamine (GalNAc) modifications, which are added via a yet unresolved process. 29374258 4 55 theme single-pass 632:642 arg1 PGAP4 567:571 PGAP4 567:571 PGAP4 567:571 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 29374258 4 55 theme single-pass 632:642 arg1 protein 653:659 a single-pass membrane protein 630:659 a single-pass membrane protein 630:659 Furthermore, we demonstrate that PGAP4, in contrast to known Golgi glycosyltransferases, is not a single-pass membrane protein but contains three transmembrane domains, including a tandem transmembrane domain insertion into its glycosyltransferase-A fold as indicated by comparative modeling. 28011641 10 0 theme Adhesion 1363:1370 arg1 experiments 1382:1392 Adhesion frequency experiments 1363:1392 Adhesion frequency experiments with a biomembrane force probe 1363:1423 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 9 1 theme Soluble 1240:1246 arg1 P-selectinE88D 1248:1261 Soluble P-selectinE88D 1240:1261 Soluble P-selectinE88D 1240:1261 Soluble P-selectinE88D bound with an ∼9-fold lower affinity to PSGL-1, a physiological ligand, due to faster dissociation. 28011641 1 2 theme fucosylated 140:150 arg1 ligands 159:165 fucosylated glycan ligands 140:165 fucosylated glycan ligands 140:165 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 5 3 theme force-assisted 715:728 arg1 allostery 730:738 force-assisted allostery 715:738 force-assisted allostery 715:738 However, transition intermediates have not been described, and the role of Glu-88 in force-assisted allostery has not been examined. 28011641 13 4 theme high 1804:1807 arg1 state 1818:1822 the high affinity state 1800:1822 the high affinity state 1800:1822 Our data support a two-state model for selectins in which Glu-88 must engage ligand to trigger allostery that stabilizes the high affinity state under force. 28011641 3 5 theme lectin/EGF 486:495 arg1 hinge 504:508 the lectin/EGF domain hinge 482:508 the lectin/EGF domain hinge 482:508 This triggers further allostery that opens the lectin/EGF domain hinge. 28011641 0 6 theme High 80:83 arg1 State 94:98 the High Affinity State 76:98 the High Affinity State 76:98 Glycan Bound to the Selectin Low Affinity State Engages Glu-88 to Stabilize the High Affinity State under Force. 28011641 1 7 with interactions 122:133 arg1 ligands 159:165 fucosylated glycan ligands 140:165 fucosylated glycan ligands 140:165 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 2 8 theme two-state 282:290 arg1 model 292:296 a two-state model 280:296 a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose 280:436 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 10 9 with interactions 1442:1453 arg1 PSGL-1 1478:1483 PSGL-1 1478:1483 PSGL-1 1478:1483 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 4 10 from bent 571:574 arg1 transition 551:560 transition 551:560 transition from the bent (low affinity) to the extended (high affinity) state 551:627 The model posits that force accelerates transition from the bent (low affinity) to the extended (high affinity) state. 28011641 13 11 theme two-state 1698:1706 arg1 model 1708:1712 a two-state model 1696:1712 a two-state model for selectins in which Glu-88 must engage ligand to trigger allostery that stabilizes the high affinity state under force 1696:1834 Our data support a two-state model for selectins in which Glu-88 must engage ligand to trigger allostery that stabilizes the high affinity state under force. 28011641 6 12 attach attached 906:913 arg1 molecule 937:944 a symmetry-related molecule 918:944 a symmetry-related molecule 918:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 12 attach attached 906:913 arg2 N-glycan 897:904 an N-glycan 894:904 an N-glycan attached to a symmetry-related molecule 894:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 13 theme L-selectin 825:834 arg1 L-selectin 825:834 L-selectin 825:834 L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule 825:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 13 theme L-selectin 825:834 arg1 domains 814:820 the lectin and EGF domains 795:820 domains 814:820 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 8 14 theme WT 1174:1175 arg1 selectins 1177:1185 WT selectins 1174:1185 WT selectins 1174:1185 We used three complementary assays to compare ligand binding to WT selectins and to E88D selectins that replaced Glu-88 with Asp. 28011641 2 15 theme coordination 377:388 arg1 site 390:393 the Ca2+ coordination site 368:393 the Ca2+ coordination site 368:393 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 0 16 theme Affinity 85:92 arg1 State 94:98 the High Affinity State 76:98 the High Affinity State 76:98 Glycan Bound to the Selectin Low Affinity State Engages Glu-88 to Stabilize the High Affinity State under Force. 28011641 10 17 theme biomembrane 1401:1411 arg1 probe 1419:1423 a biomembrane force probe 1399:1423 a biomembrane force probe 1399:1423 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 2 18 theme E-selectin 261:270 arg1 structures 240:249 Crystal structures 232:249 Crystal structures of P- and E-selectin 232:270 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 2 19 theme Ca2+ 372:375 arg1 site 390:393 the Ca2+ coordination site 368:393 the Ca2+ coordination site 368:393 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 7 20 theme domain 1096:1101 arg1 hinge 1103:1107 the lectin/EGF domain hinge 1081:1107 the lectin/EGF domain hinge 1081:1107 The structure is a transition intermediate where loop 83-89 closes to engage Ca2+ and mannose without triggering allostery that opens the lectin/EGF domain hinge. 28011641 6 21 theme domains 814:820 arg1 structure 782:790 the structure 778:790 the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule 778:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 2 22 theme ligand 307:312 arg1 binding 314:320 ligand binding 307:320 ligand binding to the lectin domain 307:341 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 1 23 theme leukocyte 175:183 arg1 rolling 185:191 leukocyte rolling 175:191 leukocyte rolling 175:191 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 10 24 theme force 1413:1417 arg1 probe 1419:1423 a biomembrane force probe 1399:1423 a biomembrane force probe 1399:1423 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 10 25 theme frequency 1372:1380 arg1 experiments 1382:1392 Adhesion frequency experiments 1363:1392 Adhesion frequency experiments with a biomembrane force probe 1363:1423 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 10 26 with experiments 1382:1392 arg1 probe 1419:1423 a biomembrane force probe 1399:1423 a biomembrane force probe 1399:1423 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 8 27 used used 1113:1116 arg2 We 1110:1111 We 1110:1111 We 1110:1111 We used three complementary assays to compare ligand binding to WT selectins and to E88D selectins that replaced Glu-88 with Asp. 28011641 9 28 theme faster 1342:1347 arg1 dissociation 1349:1360 faster dissociation 1342:1360 faster dissociation 1342:1360 Soluble P-selectinE88D bound with an ∼9-fold lower affinity to PSGL-1, a physiological ligand, due to faster dissociation. 28011641 4 29 theme extended 598:605 arg1 state 623:627 the extended (high affinity) state 594:627 the extended (high affinity) state 594:627 The model posits that force accelerates transition from the bent (low affinity) to the extended (high affinity) state. 28011641 7 30 theme lectin/EGF 1085:1094 arg1 hinge 1103:1107 the lectin/EGF domain hinge 1081:1107 the lectin/EGF domain hinge 1081:1107 The structure is a transition intermediate where loop 83-89 closes to engage Ca2+ and mannose without triggering allostery that opens the lectin/EGF domain hinge. 28011641 6 31 theme terminal 874:881 arg1 fucose 847:852 a fucose 845:852 a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule 845:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 31 theme terminal 874:881 arg1 mannose 883:889 a terminal mannose 872:889 a terminal mannose on an N-glycan attached to a symmetry-related molecule 872:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 32 theme EGF 810:812 arg1 L-selectin 825:834 L-selectin 825:834 L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule 825:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 32 theme EGF 810:812 arg1 domains 814:820 the lectin and EGF domains 795:820 domains 814:820 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 1 33 theme glycan 152:157 arg1 ligands 159:165 fucosylated glycan ligands 140:165 fucosylated glycan ligands 140:165 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 13 34 theme affinity 1809:1816 arg1 state 1818:1822 the high affinity state 1800:1822 the high affinity state 1800:1822 Our data support a two-state model for selectins in which Glu-88 must engage ligand to trigger allostery that stabilizes the high affinity state under force. 28011641 0 35 theme Low 29:31 arg1 State 42:46 the Selectin Low Affinity State 16:46 the Selectin Low Affinity State 16:46 Glycan Bound to the Selectin Low Affinity State Engages Glu-88 to Stabilize the High Affinity State under Force. 28011641 7 36 theme transition 966:975 arg1 intermediate 977:988 a transition intermediate 964:988 a transition intermediate where loop 83-89 closes to engage Ca2+ and mannose without triggering allostery that opens the lectin/EGF domain hinge 964:1107 The structure is a transition intermediate where loop 83-89 closes to engage Ca2+ and mannose without triggering allostery that opens the lectin/EGF domain hinge. 28011641 7 36 theme transition 966:975 arg1 structure 951:959 The structure 947:959 The structure 947:959 The structure is a transition intermediate where loop 83-89 closes to engage Ca2+ and mannose without triggering allostery that opens the lectin/EGF domain hinge. 28011641 5 37 from role 697:700 arg1 allostery 730:738 force-assisted allostery 715:738 force-assisted allostery 715:738 However, transition intermediates have not been described, and the role of Glu-88 in force-assisted allostery has not been examined. 28011641 2 38 theme Crystal 232:238 arg1 structures 240:249 Crystal structures 232:249 Crystal structures of P- and E-selectin 232:270 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 4 39 theme high 608:611 arg1 affinity 613:620 high affinity 608:620 high affinity 608:620 The model posits that force accelerates transition from the bent (low affinity) to the extended (high affinity) state. 28011641 0 40 theme Selectin 20:27 arg1 State 42:46 the Selectin Low Affinity State 16:46 the Selectin Low Affinity State 16:46 Glycan Bound to the Selectin Low Affinity State Engages Glu-88 to Stabilize the High Affinity State under Force. 28011641 6 41 from mannose 883:889 arg1 N-glycan 897:904 an N-glycan 894:904 an N-glycan attached to a symmetry-related molecule 894:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 42 theme lectin 799:804 arg1 L-selectin 825:834 L-selectin 825:834 L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule 825:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 42 theme lectin 799:804 arg1 domains 814:820 the lectin and EGF domains 795:820 domains 814:820 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 10 43 theme P-selectinE88D 1458:1471 arg1 interactions 1442:1453 interactions 1442:1453 interactions of P-selectinE88D with PSGL-1 1442:1483 Adhesion frequency experiments with a biomembrane force probe could not detect interactions of P-selectinE88D with PSGL-1. 28011641 9 44 theme lower 1285:1289 arg1 affinity 1291:1298 an ∼9-fold lower affinity 1274:1298 an ∼9-fold lower affinity to PSGL-1, a physiological ligand, 1274:1333 Soluble P-selectinE88D bound with an ∼9-fold lower affinity to PSGL-1, a physiological ligand, due to faster dissociation. 28011641 8 45 theme complementary 1124:1136 arg1 assays 1138:1143 three complementary assays 1118:1143 three complementary assays 1118:1143 We used three complementary assays to compare ligand binding to WT selectins and to E88D selectins that replaced Glu-88 with Asp. 28011641 4 46 dep extended 598:605 arg1 affinity 613:620 high affinity 608:620 high affinity 608:620 The model posits that force accelerates transition from the bent (low affinity) to the extended (high affinity) state. 28011641 2 47 theme lectin 329:334 arg1 domain 336:341 the lectin domain 325:341 the lectin domain 325:341 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 0 48 theme Affinity 33:40 arg1 State 42:46 the Selectin Low Affinity State 16:46 the Selectin Low Affinity State 16:46 Glycan Bound to the Selectin Low Affinity State Engages Glu-88 to Stabilize the High Affinity State under Force. 28011641 5 49 theme transition 639:648 arg1 intermediates 650:662 transition intermediates 639:662 transition intermediates 639:662 However, transition intermediates have not been described, and the role of Glu-88 in force-assisted allostery has not been examined. 28011641 11 50 theme transmembrane 1503:1515 arg1 P-selectinE88D 1517:1530 transmembrane P-selectinE88D 1503:1530 transmembrane P-selectinE88D 1503:1530 Cells expressing transmembrane P-selectinE88D or L-selectinE88D detached from immobilized ligands immediately after initiating flow. 28011641 3 51 theme further 453:459 arg1 allostery 461:469 further allostery 453:469 further allostery that opens the lectin/EGF domain hinge 453:508 This triggers further allostery that opens the lectin/EGF domain hinge. 28011641 1 52 theme shear 218:222 arg1 forces 224:229 shear forces 218:229 shear forces 218:229 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 1 53 gly fucosylated 140:150 arg1 ligands 159:165 fucosylated glycan ligands 140:165 fucosylated glycan ligands 140:165 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 2 54 theme P- 254:255 arg1 structures 240:249 Crystal structures 232:249 Crystal structures of P- and E-selectin 232:270 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 1 55 theme Selectin 113:120 arg1 interactions 122:133 Selectin interactions 113:133 Selectin interactions with fucosylated glycan ligands 113:165 Selectin interactions with fucosylated glycan ligands mediate leukocyte rolling in the vasculature under shear forces. 28011641 4 56 theme low 577:579 arg1 bent 571:574 bent 571:574 bent 571:574 The model posits that force accelerates transition from the bent (low affinity) to the extended (high affinity) state. 28011641 4 56 theme low 577:579 arg1 affinity 581:588 low affinity 577:588 low affinity 577:588 The model posits that force accelerates transition from the bent (low affinity) to the extended (high affinity) state. 28011641 3 57 theme domain 497:502 arg1 hinge 504:508 the lectin/EGF domain hinge 482:508 the lectin/EGF domain hinge 482:508 This triggers further allostery that opens the lectin/EGF domain hinge. 28011641 8 58 theme ligand 1156:1161 arg1 binding 1163:1169 ligand binding 1156:1169 ligand binding to WT selectins and to E88D selectins that replaced Glu-88 with Asp 1156:1237 We used three complementary assays to compare ligand binding to WT selectins and to E88D selectins that replaced Glu-88 with Asp. 28011641 6 59 theme symmetry-related 920:935 arg1 molecule 937:944 a symmetry-related molecule 918:944 a symmetry-related molecule 918:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 5 60 theme Glu-88 705:710 arg1 role 697:700 the role 693:700 the role of Glu-88 in force-assisted allostery 693:738 However, transition intermediates have not been described, and the role of Glu-88 in force-assisted allostery has not been examined. 28011641 6 61 theme mimetic 854:860 arg1 mannose 883:889 a terminal mannose 872:889 a terminal mannose on an N-glycan attached to a symmetry-related molecule 872:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 6 61 theme mimetic 854:860 arg1 fucose 847:852 a fucose 845:852 a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule 845:944 Here we report the structure of the lectin and EGF domains of L-selectin bound to a fucose mimetic; that is, a terminal mannose on an N-glycan attached to a symmetry-related molecule. 28011641 8 62 theme E88D 1194:1197 arg1 selectins 1199:1207 E88D selectins 1194:1207 E88D selectins that replaced Glu-88 with Asp 1194:1237 We used three complementary assays to compare ligand binding to WT selectins and to E88D selectins that replaced Glu-88 with Asp. 28011641 11 63 theme immobilized 1564:1574 arg1 ligands 1576:1582 immobilized ligands 1564:1582 immobilized ligands 1564:1582 Cells expressing transmembrane P-selectinE88D or L-selectinE88D detached from immobilized ligands immediately after initiating flow. 28011641 2 64 theme loop 350:353 arg1 83-89 355:359 loop 83-89 350:359 loop 83-89 350:359 Crystal structures of P- and E-selectin suggest a two-state model in which ligand binding to the lectin domain closes loop 83-89 around the Ca2+ coordination site, enabling Glu-88 to engage Ca2+ and fucose. 28011641 9 65 theme physiological 1313:1325 arg1 PSGL-1 1303:1308 PSGL-1 1303:1308 PSGL-1 1303:1308 Soluble P-selectinE88D bound with an ∼9-fold lower affinity to PSGL-1, a physiological ligand, due to faster dissociation. 28011641 9 65 theme physiological 1313:1325 arg1 ligand 1327:1332 a physiological ligand 1311:1332 a physiological ligand 1311:1332 Soluble P-selectinE88D bound with an ∼9-fold lower affinity to PSGL-1, a physiological ligand, due to faster dissociation. 32111832 3 0 from ablation 418:425 arg1 cells 435:439 Hap1 cells 430:439 Hap1 cells 430:439 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 0 from ablation 418:425 arg1 hepatocytes 485:495 primary murine hepatocytes 470:495 primary murine hepatocytes 470:495 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 0 from ablation 418:425 arg1 cells 459:463 Hepa1-6 hepatoma cells 442:463 Hepa1-6 hepatoma cells 442:463 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 4 1 from mice 525:528 arg1 lethal 560:565 lethal 560:565 lethal 560:565 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 7 2 theme SREBP 1013:1017 arg1 role 1005:1008 the role 1001:1008 the role of SREBP in tumor growth 1001:1033 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 8 3 theme unrecognized 1160:1171 arg1 modulator 1173:1181 a previously unrecognized modulator 1147:1181 a previously unrecognized modulator of SREBP signaling 1147:1200 In conclusion, we identify SPRING as a previously unrecognized modulator of SREBP signaling. 32111832 7 4 theme tumor 1052:1056 arg1 lines 1063:1067 tumor cell lines 1052:1067 tumor cell lines 1052:1067 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 4 5 theme hepatic 584:590 arg1 expression 599:608 hepatic Spring expression 584:608 hepatic Spring expression 584:608 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 0 6 theme signaling 75:83 arg1 SPRING/C12ORF49 33:47 SPRING/C12ORF49 33:47 SPRING/C12ORF49 33:47 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 0 6 theme signaling 75:83 arg1 determinant 54:64 a determinant 52:64 a determinant of SREBP signaling and cholesterol metabolism 52:110 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 5 7 theme SREBP 674:678 arg1 signaling 680:688 attenuated SREBP signaling 663:688 attenuated SREBP signaling in SPRINGKO cells 663:706 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 7 8 from role 1005:1008 arg1 growth 1028:1033 tumor growth 1022:1033 tumor growth 1022:1033 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 5 9 theme attenuated 663:672 arg1 signaling 680:688 attenuated SREBP signaling 663:688 attenuated SREBP signaling in SPRINGKO cells 663:706 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 0 10 theme metabolism 101:110 arg1 SPRING/C12ORF49 33:47 SPRING/C12ORF49 33:47 SPRING/C12ORF49 33:47 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 0 10 theme metabolism 101:110 arg1 determinant 54:64 a determinant 52:64 a determinant of SREBP signaling and cholesterol metabolism 52:110 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 6 11 theme SREBP-dependent 938:952 arg1 signaling 954:962 SREBP-dependent signaling 938:962 SREBP-dependent signaling 938:962 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 4 12 theme expression 599:608 arg1 silencing 571:579 silencing 571:579 silencing of hepatic Spring expression 571:608 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 0 13 theme cholesterol 89:99 arg1 metabolism 101:110 cholesterol metabolism 89:110 cholesterol metabolism 89:110 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 4 14 theme Spring 592:597 arg1 expression 599:608 hepatic Spring expression 584:608 hepatic Spring expression 584:608 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 8 15 theme SREBP 1186:1190 arg1 signaling 1192:1200 SREBP signaling 1186:1200 SREBP signaling 1186:1200 In conclusion, we identify SPRING as a previously unrecognized modulator of SREBP signaling. 32111832 6 16 with Consistent 851:860 arg1 SCAP 886:889 limited functional SCAP 867:889 limited functional SCAP in SPRINGKO cells 867:907 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 3 17 theme primary 470:476 arg1 hepatocytes 485:495 primary murine hepatocytes 470:495 primary murine hepatocytes 470:495 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 18 theme hepatoma 450:457 arg1 cells 459:463 Hepa1-6 hepatoma cells 442:463 Hepa1-6 hepatoma cells 442:463 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 1 19 theme central 172:178 arg1 regulators 196:205 central transcriptional regulators 172:205 central transcriptional regulators of lipid metabolism 172:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 19 theme central 172:178 arg1 proteins 151:158 The sterol-regulatory element binding proteins 113:158 The sterol-regulatory element binding proteins (SREBP) 113:166 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 6 20 theme reintroducing 910:922 arg1 SCAP 924:927 reintroducing SCAP 910:927 reintroducing SCAP 910:927 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 5 21 theme sterol 836:841 arg1 status 843:848 the cellular sterol status 823:848 the cellular sterol status 823:848 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 1 22 theme transcriptional 180:194 arg1 regulators 196:205 central transcriptional regulators 172:205 central transcriptional regulators of lipid metabolism 172:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 22 theme transcriptional 180:194 arg1 proteins 151:158 The sterol-regulatory element binding proteins 113:158 The sterol-regulatory element binding proteins (SREBP) 113:166 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 0 23 theme genetic 8:14 arg1 screens 16:22 Haploid genetic screens 0:22 Haploid genetic screens 0:22 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 8 24 theme signaling 1192:1200 arg1 modulator 1173:1181 a previously unrecognized modulator 1147:1181 a previously unrecognized modulator of SREBP signaling 1147:1200 In conclusion, we identify SPRING as a previously unrecognized modulator of SREBP signaling. 32111832 7 25 theme SPRING 1091:1096 arg1 expression 1098:1107 SPRING expression 1091:1107 SPRING expression 1091:1107 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 7 26 theme cell 1058:1061 arg1 lines 1063:1067 tumor cell lines 1052:1067 tumor cell lines 1052:1067 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 0 27 theme Haploid 0:6 arg1 screens 16:22 Haploid genetic screens 0:22 Haploid genetic screens 0:22 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 6 28 from SCAP 886:889 arg1 cells 903:907 SPRINGKO cells 894:907 SPRINGKO cells 894:907 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 3 29 theme Golgi-resident 378:391 arg1 protein 402:408 a glycosylated Golgi-resident membrane protein 363:408 a glycosylated Golgi-resident membrane protein 363:408 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 29 theme Golgi-resident 378:391 arg1 SPRING 353:358 SPRING 353:358 SPRING 353:358 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 30 gly glycosylated 365:376 arg1 protein 402:408 a glycosylated Golgi-resident membrane protein 363:408 a glycosylated Golgi-resident membrane protein 363:408 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 30 gly glycosylated 365:376 arg1 SPRING 353:358 SPRING 353:358 SPRING 353:358 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 1 31 theme lipid 210:214 arg1 metabolism 216:225 lipid metabolism 210:225 lipid metabolism 210:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 3 32 theme membrane 393:400 arg1 protein 402:408 a glycosylated Golgi-resident membrane protein 363:408 a glycosylated Golgi-resident membrane protein 363:408 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 32 theme membrane 393:400 arg1 SPRING 353:358 SPRING 353:358 SPRING 353:358 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 7 33 theme lines 1063:1067 arg1 range 1043:1047 a wide range 1036:1047 a wide range of tumor cell lines 1036:1067 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 5 34 theme SREBP 729:733 arg1 SCAP 764:767 SCAP 764:767 SCAP 764:767 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 5 34 theme SREBP 729:733 arg1 protein 755:761 reduced SREBP cleavage-activating protein 721:761 reduced SREBP cleavage-activating protein (SCAP) 721:768 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 3 35 theme SREBP 505:509 arg1 signaling 511:519 SREBP signaling 505:519 SREBP signaling 505:519 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 1 36 theme metabolism 216:225 arg1 regulators 196:205 central transcriptional regulators 172:205 central transcriptional regulators of lipid metabolism 172:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 36 theme metabolism 216:225 arg1 proteins 151:158 The sterol-regulatory element binding proteins 113:158 The sterol-regulatory element binding proteins (SREBP) 113:166 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 2 37 theme pathway 344:350 arg1 Gene 291:294 the SREBP Regulating Gene 270:294 the SREBP Regulating Gene (SPRING/C12ORF49) 270:312 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 2 37 theme pathway 344:350 arg1 determinant 319:329 a determinant 317:329 a determinant of the SREBP pathway 317:350 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 5 38 theme cleavage-activating 735:753 arg1 SCAP 764:767 SCAP 764:767 SCAP 764:767 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 5 38 theme cleavage-activating 735:753 arg1 protein 755:761 reduced SREBP cleavage-activating protein 721:761 reduced SREBP cleavage-activating protein (SCAP) 721:768 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 6 39 theme SPRINGKO 894:901 arg1 cells 903:907 SPRINGKO cells 894:907 SPRINGKO cells 894:907 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 3 40 theme Hap1 430:433 arg1 cells 435:439 Hap1 cells 430:439 Hap1 cells 430:439 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 4 41 theme Spring 531:536 arg1 deletion 538:545 Spring deletion 531:545 Spring deletion 531:545 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 5 42 theme irrespective 807:818 arg1 Golgi 801:805 the Golgi 797:805 the Golgi irrespective of the cellular sterol status 797:848 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 4 43 from lethal 560:565 arg1 mice 525:528 mice 525:528 mice 525:528 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 5 44 theme status 843:848 arg1 irrespective 807:818 irrespective 807:818 irrespective 807:818 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 5 45 from signaling 680:688 arg1 cells 702:706 SPRINGKO cells 693:706 SPRINGKO cells 693:706 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 5 46 theme SPRINGKO 693:700 arg1 cells 702:706 SPRINGKO cells 693:706 SPRINGKO cells 693:706 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 3 47 theme Hepa1-6 442:448 arg1 cells 459:463 Hepa1-6 hepatoma cells 442:463 Hepa1-6 hepatoma cells 442:463 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 2 48 theme genetic 242:248 arg1 screens 250:256 haploid genetic screens 234:256 haploid genetic screens 234:256 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 7 49 theme wide 1038:1041 arg1 range 1043:1047 a wide range 1036:1047 a wide range of tumor cell lines 1036:1067 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 1 50 theme sterol-regulatory 117:133 arg1 regulators 196:205 central transcriptional regulators 172:205 central transcriptional regulators of lipid metabolism 172:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 50 theme sterol-regulatory 117:133 arg1 SREBP 161:165 SREBP 161:165 SREBP 161:165 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 50 theme sterol-regulatory 117:133 arg1 proteins 151:158 The sterol-regulatory element binding proteins 113:158 The sterol-regulatory element binding proteins (SREBP) 113:166 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 3 51 theme murine 478:483 arg1 hepatocytes 485:495 primary murine hepatocytes 470:495 primary murine hepatocytes 470:495 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 6 52 theme functional 875:884 arg1 SCAP 886:889 limited functional SCAP 867:889 limited functional SCAP in SPRINGKO cells 867:907 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 2 53 theme haploid 234:240 arg1 screens 250:256 haploid genetic screens 234:256 haploid genetic screens 234:256 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 1 54 theme element 135:141 arg1 regulators 196:205 central transcriptional regulators 172:205 central transcriptional regulators of lipid metabolism 172:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 54 theme element 135:141 arg1 SREBP 161:165 SREBP 161:165 SREBP 161:165 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 54 theme element 135:141 arg1 proteins 151:158 The sterol-regulatory element binding proteins 113:158 The sterol-regulatory element binding proteins (SREBP) 113:166 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 7 55 with line 991:994 arg1 role 1005:1008 the role 1001:1008 the role of SREBP in tumor growth 1001:1033 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 6 56 theme limited 867:873 arg1 SCAP 886:889 limited functional SCAP 867:889 limited functional SCAP in SPRINGKO cells 867:907 Consistent with limited functional SCAP in SPRINGKO cells, reintroducing SCAP restores SREBP-dependent signaling and function. 32111832 2 57 theme Regulating 280:289 arg1 SPRING/C12ORF49 297:311 SPRING/C12ORF49 297:311 SPRING/C12ORF49 297:311 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 2 57 theme Regulating 280:289 arg1 Gene 291:294 the SREBP Regulating Gene 270:294 the SREBP Regulating Gene (SPRING/C12ORF49) 270:312 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 2 57 theme Regulating 280:289 arg1 determinant 319:329 a determinant 317:329 a determinant of the SREBP pathway 317:350 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 1 58 theme binding 143:149 arg1 regulators 196:205 central transcriptional regulators 172:205 central transcriptional regulators of lipid metabolism 172:225 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 58 theme binding 143:149 arg1 SREBP 161:165 SREBP 161:165 SREBP 161:165 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 1 58 theme binding 143:149 arg1 proteins 151:158 The sterol-regulatory element binding proteins 113:158 The sterol-regulatory element binding proteins (SREBP) 113:166 The sterol-regulatory element binding proteins (SREBP) are central transcriptional regulators of lipid metabolism. 32111832 2 59 theme SREBP 274:278 arg1 SPRING/C12ORF49 297:311 SPRING/C12ORF49 297:311 SPRING/C12ORF49 297:311 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 2 59 theme SREBP 274:278 arg1 Gene 291:294 the SREBP Regulating Gene 270:294 the SREBP Regulating Gene (SPRING/C12ORF49) 270:312 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 2 59 theme SREBP 274:278 arg1 determinant 319:329 a determinant 317:329 a determinant of the SREBP pathway 317:350 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 2 60 theme SREBP 338:342 arg1 pathway 344:350 the SREBP pathway 334:350 the SREBP pathway 334:350 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 0 61 theme SREBP 69:73 arg1 signaling 75:83 SREBP signaling 69:83 SREBP signaling 69:83 Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism. 32111832 7 62 theme tumor 1022:1026 arg1 growth 1028:1033 tumor growth 1022:1033 tumor growth 1022:1033 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 2 63 dep identify 261:268 arg1 Using 228:232 Using 228:232 Using haploid genetic screens 228:256 Using haploid genetic screens we identify the SREBP Regulating Gene (SPRING/C12ORF49) as a determinant of the SREBP pathway. 32111832 4 64 theme SREBP 630:634 arg1 response 636:643 the SREBP response 626:643 the SREBP response 626:643 In mice, Spring deletion is embryonic lethal yet silencing of hepatic Spring expression also attenuates the SREBP response. 32111832 7 65 from dependency 1077:1086 arg1 expression 1098:1107 SPRING expression 1091:1107 SPRING expression 1091:1107 Moreover, in line with the role of SREBP in tumor growth, a wide range of tumor cell lines display dependency on SPRING expression. 32111832 5 66 theme reduced 721:727 arg1 SCAP 764:767 SCAP 764:767 SCAP 764:767 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 5 66 theme reduced 721:727 arg1 protein 755:761 reduced SREBP cleavage-activating protein 721:761 reduced SREBP cleavage-activating protein (SCAP) 721:768 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 5 67 theme cellular 827:834 arg1 status 843:848 the cellular sterol status 823:848 the cellular sterol status 823:848 Mechanistically, attenuated SREBP signaling in SPRINGKO cells results from reduced SREBP cleavage-activating protein (SCAP) and its mislocalization to the Golgi irrespective of the cellular sterol status. 32111832 3 68 theme glycosylated 365:376 arg1 protein 402:408 a glycosylated Golgi-resident membrane protein 363:408 a glycosylated Golgi-resident membrane protein 363:408 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 32111832 3 68 theme glycosylated 365:376 arg1 SPRING 353:358 SPRING 353:358 SPRING 353:358 SPRING is a glycosylated Golgi-resident membrane protein and its ablation in Hap1 cells, Hepa1-6 hepatoma cells, and primary murine hepatocytes reduces SREBP signaling. 11285216 6 0 dep compact 852:858 arg1 globular 861:868 globular 861:868 globular 861:868 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 1 1 from region 183:188 arg1 fragment 149:156 the (6)F1(1)F2(2)F2 fragment 129:156 the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 129:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 1 1 from region 183:188 arg1 structure 116:124 The solution structure 103:124 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 103:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 3 2 theme intermodule 470:480 arg1 interface 482:490 the largest intermodule interface 458:490 the largest intermodule interface seen in fibronectin to date 458:518 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 3 2 theme intermodule 470:480 arg1 area 400:403 The buried surface area 381:403 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) 381:453 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 2 3 theme non-contiguous 341:354 arg1 F1 359:360 the non-contiguous (6)F1 337:360 the non-contiguous (6)F1 337:360 The structure reveals an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules. 11285216 5 4 theme intramolecular 704:717 arg1 contact 719:725 intramolecular contact 704:725 intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain 704:797 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 5 5 theme F2 686:687 arg1 topology 661:668 The hairpin topology 649:668 The hairpin topology of (6)F1(1)F2(2)F2 649:687 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 1 6 theme F2 141:142 arg1 fragment 149:156 the (6)F1(1)F2(2)F2 fragment 129:156 the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 129:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 2 7 dep F2 369:370 arg1 2 367:367 2 367:367 2 367:367 The structure reveals an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules. 11285216 6 8 theme first 812:816 arg1 This 800:803 This 800:803 This 800:803 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 6 8 theme first 812:816 arg1 study 834:838 the first high-resolution study 808:838 the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin 808:906 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 7 9 theme linear 987:992 arg1 fibronectin 963:973 fibronectin 963:973 fibronectin 963:973 This arrangement is not consistent with the view that fibronectin is simply a linear 'string of beads'. 11285216 7 9 theme linear 987:992 arg1 string 995:1000 a linear 'string 985:1000 a linear 'string of beads' 985:1010 This arrangement is not consistent with the view that fibronectin is simply a linear 'string of beads'. 11285216 4 10 theme near-complete 601:613 arg1 loss 615:618 near-complete loss 601:618 near-complete loss of gelatin-binding activity 601:646 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 0 11 theme F2 37:38 arg1 fragment 45:52 the (6)F1(1)F2(2)F2 fragment 25:52 the (6)F1(1)F2(2)F2 fragment 25:52 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 0 12 from fibronectin 65:75 arg1 structure 12:20 The hairpin structure 0:20 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin 0:75 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 4 13 theme F2 587:588 arg1 dissection 525:534 The dissection 521:534 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 521:588 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 1 14 theme Protein 226:232 arg1 Bank 239:242 Protein Data Bank 226:242 Protein Data Bank entry 226:248 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 6 15 theme compact 852:858 arg1 arrangement 870:880 a compact, globular arrangement 850:880 a compact, globular arrangement of modules in fibronectin 850:906 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 4 16 dep F1 542:543 arg1 1 545:545 1 545:545 1 545:545 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 4 16 dep F1 542:543 arg1 F2 552:553 F2 552:553 F2 552:553 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 4 16 dep F1 542:543 arg1 2 550:550 2 550:550 2 550:550 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 2 17 theme F1 359:360 arg1 modules 372:378 the non-contiguous (6)F1 and (2)F2 modules 337:378 the non-contiguous (6)F1 and (2)F2 modules 337:378 The structure reveals an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules. 11285216 1 18 theme Data 234:237 arg1 Bank 239:242 Protein Data Bank 226:242 Protein Data Bank entry 226:248 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 3 19 dep F2 426:427 arg1 2 424:424 2 424:424 2 424:424 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 0 20 theme hairpin 4:10 arg1 structure 12:20 The hairpin structure 0:20 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin 0:75 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 1 21 theme Bank 239:242 arg1 entry 244:248 Protein Data Bank entry 226:248 Protein Data Bank entry 226:248 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 3 22 theme surface 392:398 arg1 interface 482:490 the largest intermodule interface 458:490 the largest intermodule interface seen in fibronectin to date 458:518 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 3 22 theme surface 392:398 arg1 area 400:403 The buried surface area 381:403 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) 381:453 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 1 23 theme solution 107:114 arg1 structure 116:124 The solution structure 103:124 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 103:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 2 24 theme F2 369:370 arg1 modules 372:378 the non-contiguous (6)F1 and (2)F2 modules 337:378 the non-contiguous (6)F1 and (2)F2 modules 337:378 The structure reveals an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules. 11285216 1 25 theme F2 146:147 arg1 fragment 149:156 the (6)F1(1)F2(2)F2 fragment 129:156 the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 129:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 0 26 theme fragment 45:52 arg1 structure 12:20 The hairpin structure 0:20 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin 0:75 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 4 27 dep F2 587:588 arg1 2 585:585 2 585:585 2 585:585 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 1 28 theme fragment 149:156 arg1 structure 116:124 The solution structure 103:124 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 103:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 0 29 theme F2 42:43 arg1 fragment 45:52 the (6)F1(1)F2(2)F2 fragment 25:52 the (6)F1(1)F2(2)F2 fragment 25:52 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 3 30 theme buried 385:390 arg1 interface 482:490 the largest intermodule interface 458:490 the largest intermodule interface seen in fibronectin to date 458:518 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 3 30 theme buried 385:390 arg1 area 400:403 The buried surface area 381:403 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) 381:453 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 4 31 theme gelatin-binding 623:637 arg1 activity 639:646 gelatin-binding activity 623:646 gelatin-binding activity 623:646 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 5 32 theme gelatin-binding 776:790 arg1 domain 792:797 the gelatin-binding domain 772:797 the gelatin-binding domain 772:797 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 0 33 theme human 59:63 arg1 fibronectin 65:75 human fibronectin 59:75 human fibronectin 59:75 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 4 34 theme activity 639:646 arg1 loss 615:618 near-complete loss 601:618 near-complete loss of gelatin-binding activity 601:646 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 5 35 theme hairpin 653:659 arg1 topology 661:668 The hairpin topology 649:668 The hairpin topology of (6)F1(1)F2(2)F2 649:687 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 6 36 theme modules 885:891 arg1 arrangement 870:880 a compact, globular arrangement 850:880 a compact, globular arrangement of modules in fibronectin 850:906 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 1 37 theme gelatin-binding 167:181 arg1 region 183:188 the gelatin-binding region 163:188 the gelatin-binding region of fibronectin 163:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 4 38 theme F1 567:568 arg1 pair 575:578 the (6)F1(1)F2 pair 560:578 the (6)F1(1)F2 pair 560:578 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 5 39 theme F2 681:682 arg1 F2 686:687 (6)F1(1)F2(2)F2 673:687 (6)F1(1)F2(2)F2 673:687 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 4 40 theme F2 547:548 arg1 F1 542:543 (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair 539:578 (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair 539:578 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 3 41 theme largest 462:468 arg1 interface 482:490 the largest intermodule interface 458:490 the largest intermodule interface seen in fibronectin to date 458:518 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 3 41 theme largest 462:468 arg1 area 400:403 The buried surface area 381:403 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) 381:453 The buried surface area between (6)F1 and (2)F2 ( approximately 870 A(2)) is the largest intermodule interface seen in fibronectin to date. 11285216 2 42 theme hydrophobic 307:317 arg1 interface 319:327 an extensive hydrophobic interface 294:327 an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules 294:378 The structure reveals an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules. 11285216 5 43 theme matrix 739:744 arg1 regions 755:761 the matrix assembly regions 735:761 the matrix assembly regions flanking the gelatin-binding domain 735:797 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 4 44 theme F2 572:573 arg1 pair 575:578 the (6)F1(1)F2 pair 560:578 the (6)F1(1)F2 pair 560:578 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 5 45 theme assembly 746:753 arg1 regions 755:761 the matrix assembly regions 735:761 the matrix assembly regions flanking the gelatin-binding domain 735:797 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 6 46 theme high-resolution 818:832 arg1 This 800:803 This 800:803 This 800:803 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 6 46 theme high-resolution 818:832 arg1 study 834:838 the first high-resolution study 808:838 the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin 808:906 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 0 47 theme gelatin 86:92 arg1 binding 94:100 gelatin binding 86:100 gelatin binding 86:100 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 1 48 theme F1 136:137 arg1 fragment 149:156 the (6)F1(1)F2(2)F2 fragment 129:156 the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin 129:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 7 49 with consistent 933:942 arg1 view 953:956 the view that fibronectin is simply a linear 'string of beads' 949:1010 the view that fibronectin is simply a linear 'string of beads' 949:1010 This arrangement is not consistent with the view that fibronectin is simply a linear 'string of beads'. 11285216 5 50 theme F1 676:677 arg1 F2 686:687 (6)F1(1)F2(2)F2 673:687 (6)F1(1)F2(2)F2 673:687 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 11285216 4 51 theme F1 542:543 arg1 dissection 525:534 The dissection 521:534 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 521:588 The dissection of (6)F1(1)F2(2)F2 into the (6)F1(1)F2 pair and (2)F2 results in near-complete loss of gelatin-binding activity. 11285216 1 52 from structure 116:124 arg1 region 183:188 the gelatin-binding region 163:188 the gelatin-binding region of fibronectin 163:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 0 53 theme F1 32:33 arg1 fragment 45:52 the (6)F1(1)F2(2)F2 fragment 25:52 the (6)F1(1)F2(2)F2 fragment 25:52 The hairpin structure of the (6)F1(1)F2(2)F2 fragment from human fibronectin enhances gelatin binding. 11285216 1 54 theme fibronectin 193:203 arg1 region 183:188 the gelatin-binding region 163:188 the gelatin-binding region of fibronectin 163:203 The solution structure of the (6)F1(1)F2(2)F2 fragment from the gelatin-binding region of fibronectin has been determined (Protein Data Bank entry codes 1e88 and 1e8b). 11285216 6 55 from arrangement 870:880 arg1 fibronectin 896:906 fibronectin 896:906 fibronectin 896:906 This is the first high-resolution study to reveal a compact, globular arrangement of modules in fibronectin. 11285216 2 56 theme extensive 297:305 arg1 interface 319:327 an extensive hydrophobic interface 294:327 an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules 294:378 The structure reveals an extensive hydrophobic interface between the non-contiguous (6)F1 and (2)F2 modules. 11285216 5 57 dep F2 686:687 arg1 2 684:684 2 684:684 2 684:684 The hairpin topology of (6)F1(1)F2(2)F2 may facilitate intramolecular contact between the matrix assembly regions flanking the gelatin-binding domain. 21752865 4 0 dep residues 826:833 arg1 170-173 835:841 170-173 835:841 170-173 835:841 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 4 1 theme consensus 794:802 arg1 sequence 804:811 the consensus sequence 790:811 the consensus sequence Asn-Thr-Ser (residues 170-173) 790:842 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 2 2 theme type 317:320 arg1 BRI2 307:310 BRI2 307:310 BRI2 307:310 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 2 2 theme type 317:320 arg1 protein 339:345 a type II transmembrane protein 315:345 a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function 315:549 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 1 3 theme British 206:212 arg1 dementias 225:233 familial British and Danish dementias 197:233 familial British and Danish dementias 197:233 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 11 4 from surface 1876:1882 arg1 expression 1853:1862 its expression 1849:1862 its expression at the cell surface 1849:1882 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 8 5 theme pulse-chase 1401:1411 arg1 experiments 1413:1423 pulse-chase experiments 1401:1423 pulse-chase experiments 1401:1423 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 6 6 theme kidney 1064:1069 arg1 cells 1072:1076 HEK293 (human embryonic kidney) cells 1040:1076 HEK293 (human embryonic kidney) cells expressing BRI2 1040:1092 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 0 7 theme cell 78:81 arg1 surface 83:89 the cell surface 74:89 the cell surface 74:89 Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10. 21752865 0 8 from Glycosylation 0:12 arg1 asparagine 25:34 asparagine 170 25:38 asparagine 170 25:38 Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10. 21752865 2 9 theme function 542:549 arg1 fragments 521:529 secreted fragments 512:529 secreted fragments of unknown function 512:549 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 9 10 theme Asn170 1468:1473 arg1 mutation 1456:1463 mutation 1456:1463 mutation of Asn170 to alanine 1456:1484 These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. 21752865 1 11 theme Danish 218:223 arg1 dementias 225:233 familial British and Danish dementias 197:233 familial British and Danish dementias 197:233 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 9 12 theme BRI2 1494:1497 arg1 trafficking 1499:1509 BRI2 trafficking 1494:1509 BRI2 trafficking 1494:1509 These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. 21752865 2 13 theme unknown 534:540 arg1 function 542:549 unknown function 534:549 unknown function 534:549 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 6 14 theme inhibitor 1119:1127 arg1 tunicamycin 1129:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 6 15 theme human 1048:1052 arg1 kidney 1064:1069 human embryonic kidney 1048:1069 HEK293 (human embryonic kidney) cells expressing BRI2 1040:1092 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 3 16 theme amino 669:673 arg1 acids 675:679 amino acids 669:679 amino acids (30 kDa) 669:688 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 3 16 theme amino 669:673 arg1 kDa 685:687 30 kDa 682:687 30 kDa 682:687 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 6 17 theme N-glycosylation 1103:1117 arg1 tunicamycin 1129:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 11 18 theme post-translational 1800:1817 arg1 modification 1819:1830 this post-translational modification 1795:1830 this post-translational modification 1795:1830 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 8 19 gly N-glycosylation 1296:1310 arg1 BRI2 1315:1318 BRI2 trafficking 1315:1330 BRI2 trafficking 1315:1330 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 11 20 gly N-glycosylated 1756:1769 arg2 Asn170 1774:1779 Asn170 1774:1779 Asn170 1774:1779 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 11 20 gly N-glycosylated 1756:1769 arg1 BRI2 1748:1751 BRI2 1748:1751 BRI2 1748:1751 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 3 21 dep number 643:648 arg1 the 639:641 the 639:641 the 639:641 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 9 22 theme plasma 1566:1571 arg1 membrane 1573:1580 the plasma membrane 1562:1580 the plasma membrane 1562:1580 These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. 21752865 8 23 theme N-glycosylation 1296:1310 arg1 effect 1286:1291 the effect 1282:1291 the effect of N-glycosylation on BRI2 trafficking 1282:1330 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 6 24 with Treatment 1027:1035 arg1 mutation 1144:1151 mutation 1144:1151 mutation of Asn170 to alanine 1144:1172 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 6 24 with Treatment 1027:1035 arg1 tunicamycin 1129:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 1 25 theme different 141:149 arg1 forms 159:163 Two different mutated forms 137:163 Two different mutated forms of BRI2 protein 137:179 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 2 26 theme cell 403:406 arg1 surface 408:414 the cell surface 399:414 the cell surface 399:414 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 6 27 theme molecular 1186:1194 arg1 mass 1196:1199 its molecular mass 1182:1199 its molecular mass 1182:1199 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 7 28 gly N-glycosylated 1245:1258 arg1 BRI2 1237:1240 BRI2 1237:1240 BRI2 1237:1240 These data indicate that BRI2 is N-glycosylated at Asn170. 21752865 7 28 gly N-glycosylated 1245:1258 arg2 Asn170 1263:1268 Asn170 1263:1268 Asn170 1263:1268 These data indicate that BRI2 is N-glycosylated at Asn170. 21752865 1 29 theme mutated 151:157 arg1 forms 159:163 Two different mutated forms 137:163 Two different mutated forms of BRI2 protein 137:179 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 11 30 from Asn170 1774:1779 arg1 N-glycosylated 1756:1769 N-glycosylated 1756:1769 N-glycosylated 1756:1769 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 5 31 theme protein 938:944 arg1 folding 946:952 protein folding 938:952 protein folding 938:952 Given that N-glycosylation is considered essential for protein folding, processing and trafficking, we examined whether BRI2 is N-glycosylated. 21752865 11 32 theme cell 1871:1874 arg1 surface 1876:1882 the cell surface 1867:1882 the cell surface 1867:1882 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 8 33 theme BRI2 1315:1318 arg1 trafficking 1320:1330 BRI2 trafficking 1315:1330 BRI2 trafficking 1315:1330 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 9 34 theme cell 1518:1521 arg1 surface 1523:1529 the cell surface 1514:1529 the cell surface 1514:1529 These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. 21752865 0 35 theme BRI2 17:20 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of BRI2 on asparagine 170 0:38 Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10. 21752865 3 36 theme acids 675:679 arg1 composition 654:664 composition 654:664 composition 654:664 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 3 36 theme acids 675:679 arg1 number 643:648 number 643:648 number 643:648 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 6 37 theme cells 1072:1076 arg1 Treatment 1027:1035 Treatment 1027:1035 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine 1027:1172 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 10 38 theme BRI2 1670:1673 arg1 cleavage 1658:1665 cleavage 1658:1665 cleavage of BRI2 by furin or ADAM10 1658:1692 Furthermore, we obtained data indicating that this mutation did not affect cleavage of BRI2 by furin or ADAM10. 21752865 2 39 theme secretory 378:386 arg1 pathway 388:394 the secretory pathway 374:394 the secretory pathway 374:394 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 4 40 dep Asn-Thr-Ser 813:823 arg1 residues 826:833 residues 170-173 826:841 residues 170-173 826:841 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 1 41 theme neuropathological 250:266 arg1 similarities 268:279 neuropathological similarities 250:279 neuropathological similarities with Alzheimer's disease 250:304 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 0 42 gly Glycosylation 0:12 arg1 trafficking 59:69 its trafficking 55:69 its trafficking to the cell surface 55:89 Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10. 21752865 0 42 gly Glycosylation 0:12 arg1 BRI2 17:20 BRI2 17:20 BRI2 17:20 Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10. 21752865 0 42 gly Glycosylation 0:12 arg1 processing 106:115 its processing 102:115 its processing by furin or ADAM10 102:134 Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10. 21752865 1 43 theme BRI2 168:171 arg1 protein 173:179 BRI2 protein 168:179 BRI2 protein 168:179 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 1 44 theme protein 173:179 arg1 forms 159:163 Two different mutated forms 137:163 Two different mutated forms of BRI2 protein 137:179 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 21752865 2 45 theme metalloproteinase 472:488 arg1 domain 490:495 metalloproteinase domain 10 472:498 metalloproteinase domain 10 472:498 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 8 46 theme cell 1339:1342 arg1 surface 1344:1350 the cell surface 1335:1350 the cell surface 1335:1350 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 11 47 theme theoretical 1719:1729 arg1 predictions 1731:1741 the theoretical predictions 1715:1741 the theoretical predictions 1715:1741 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 2 48 dep furin 436:440 arg1 disintegrin 456:466 a disintegrin 454:466 a disintegrin 454:466 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 2 48 dep furin 436:440 arg1 domain 490:495 metalloproteinase domain 10 472:498 metalloproteinase domain 10 472:498 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 8 49 theme S 1388:1388 arg1 35 1385:1386 35 1385:1386 35 1385:1386 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 8 49 theme S 1388:1388 arg1 methionine 1390:1399 S methionine 1388:1399 (35)S methionine 1384:1399 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 4 50 theme bioinformatics 740:753 arg1 analysis 755:762 bioinformatics analysis 740:762 bioinformatics analysis 740:762 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 4 51 theme Asn-Thr-Ser 813:823 arg1 sequence 804:811 the consensus sequence 790:811 the consensus sequence Asn-Thr-Ser (residues 170-173) 790:842 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 8 52 from effect 1286:1291 arg1 trafficking 1320:1330 BRI2 trafficking 1315:1330 BRI2 trafficking 1315:1330 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 8 53 dep biotinylation 1366:1378 arg1 experiments 1413:1423 pulse-chase experiments 1401:1423 pulse-chase experiments 1401:1423 To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. 21752865 2 54 theme secreted 512:519 arg1 fragments 521:529 secreted fragments 512:529 secreted fragments of unknown function 512:549 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 9 55 theme steady 1539:1544 arg1 levels 1552:1557 its steady state levels 1535:1557 its steady state levels 1535:1557 These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. 21752865 6 56 theme Asn170 1156:1161 arg1 mutation 1144:1151 mutation 1144:1151 mutation of Asn170 to alanine 1144:1172 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 6 56 theme Asn170 1156:1161 arg1 tunicamycin 1129:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 the N-glycosylation inhibitor tunicamycin 1099:1139 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 6 57 theme embryonic 1054:1062 arg1 kidney 1064:1069 human embryonic kidney 1048:1069 HEK293 (human embryonic kidney) cells expressing BRI2 1040:1092 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 4 58 contain bears 784:788 arg1 BRI2 779:782 BRI2 779:782 BRI2 779:782 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 4 58 contain bears 784:788 arg2 sequence 804:811 the consensus sequence 790:811 the consensus sequence Asn-Thr-Ser (residues 170-173) 790:842 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 3 59 gly glycosylated 714:725 arg1 BRI2 706:709 BRI2 706:709 BRI2 706:709 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 9 60 theme state 1546:1550 arg1 levels 1552:1557 its steady state levels 1535:1557 its steady state levels 1535:1557 These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. 21752865 4 61 gly N-glycosylated 857:870 arg1 BRI2 779:782 BRI2 779:782 BRI2 779:782 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 4 61 gly N-glycosylated 857:870 arg2 Asn170 875:880 Asn170 875:880 Asn170 875:880 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 3 62 theme apparent 556:563 arg1 mass 575:578 Its apparent molecular mass 552:578 Its apparent molecular mass (42-44 kDa) 552:590 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 3 62 theme apparent 556:563 arg1 kDa 587:589 42-44 kDa 581:589 42-44 kDa 581:589 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 11 63 theme proteolytic 1900:1910 arg1 processing 1912:1921 its proteolytic processing 1896:1921 its proteolytic processing 1896:1921 Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing. 21752865 2 64 theme transmembrane 325:337 arg1 BRI2 307:310 BRI2 307:310 BRI2 307:310 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 2 64 theme transmembrane 325:337 arg1 protein 339:345 a type II transmembrane protein 315:345 a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function 315:549 BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. 21752865 5 65 gly N-glycosylated 1011:1024 arg1 BRI2 1003:1006 BRI2 1003:1006 BRI2 1003:1006 Given that N-glycosylation is considered essential for protein folding, processing and trafficking, we examined whether BRI2 is N-glycosylated. 21752865 4 66 from Asn170 875:880 arg1 N-glycosylated 857:870 N-glycosylated 857:870 N-glycosylated 857:870 In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. 21752865 3 67 theme molecular 565:573 arg1 mass 575:578 Its apparent molecular mass 552:578 Its apparent molecular mass (42-44 kDa) 552:590 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 3 67 theme molecular 565:573 arg1 kDa 587:589 42-44 kDa 581:589 42-44 kDa 581:589 Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. 21752865 7 68 from Asn170 1263:1268 arg1 N-glycosylated 1245:1258 N-glycosylated 1245:1258 N-glycosylated 1245:1258 These data indicate that BRI2 is N-glycosylated at Asn170. 21752865 6 69 theme HEK293 1040:1045 arg1 cells 1072:1076 HEK293 (human embryonic kidney) cells 1040:1076 HEK293 (human embryonic kidney) cells expressing BRI2 1040:1092 Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by ~2 kDa. 21752865 1 70 theme familial 197:204 arg1 dementias 225:233 familial British and Danish dementias 197:233 familial British and Danish dementias 197:233 Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. 22846996 5 0 theme high 1055:1058 arg1 affinity 1060:1067 high affinity 1055:1067 high affinity 1055:1067 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 4 1 theme structural 809:818 arg1 homology 820:827 high structural homology 804:827 high structural homology to the B7 superfamily of proteins 804:861 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 3 2 from role 417:420 arg1 response 477:484 this response 472:484 this response 472:484 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 0 3 theme responses 58:66 arg1 modulation 24:33 modulation 24:33 modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies 24:118 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 4 4 theme high 804:807 arg1 homology 820:827 high structural homology 804:827 high structural homology to the B7 superfamily of proteins 804:861 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 5 theme proteins 854:861 arg1 superfamily 839:849 the B7 superfamily 832:849 the B7 superfamily of proteins 832:861 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 6 6 theme Vγ9Vδ2 1364:1369 arg1 receptor 1378:1385 the Vγ9Vδ2 T cell receptor 1360:1385 the Vγ9Vδ2 T cell receptor 1360:1385 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 4 7 from homodimers 885:894 arg1 solution 899:906 solution 899:906 solution 899:906 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 8 theme V-shaped 876:883 arg1 homodimers 885:894 V-shaped homodimers 876:894 V-shaped homodimers in solution 876:906 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 6 9 theme extracellular 1301:1313 arg1 events 1315:1320 the extracellular events 1297:1320 the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor 1297:1385 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 5 10 theme BTN3A 1032:1036 arg1 domain 1043:1048 the BTN3A Ig-V domain 1028:1048 the BTN3A Ig-V domain 1028:1048 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 2 11 theme population 366:375 arg1 activation 337:346 the activation 333:346 the activation of this γδ T cell population 333:375 However, the molecular mechanisms behind the activation of this γδ T cell population remains unclear. 22846996 4 12 theme C-type 951:956 arg1 domain 961:966 the membrane proximal C-type Ig domain 929:966 the membrane proximal C-type Ig domain 929:966 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 5 13 theme Ig-V 1038:1041 arg1 domain 1043:1048 the BTN3A Ig-V domain 1028:1048 the BTN3A Ig-V domain 1028:1048 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 6 14 theme cell 1373:1376 arg1 receptor 1378:1385 the Vγ9Vδ2 T cell receptor 1360:1385 the Vγ9Vδ2 T cell receptor 1360:1385 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 2 15 theme cell 361:364 arg1 population 366:375 this γδ T cell population 351:375 this γδ T cell population 351:375 However, the molecular mechanisms behind the activation of this γδ T cell population remains unclear. 22846996 0 16 theme molecular 4:12 arg1 basis 14:18 The molecular basis 0:18 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 0:119 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 5 17 theme binding 1120:1126 arg1 orientation 1128:1138 their binding orientation 1114:1138 their binding orientation 1114:1138 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 3 18 theme molecules 532:540 arg1 structures 512:521 the structures 508:521 the structures of these molecules 508:540 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 3 18 theme molecules 532:540 arg1 modifications 552:564 their modifications 546:564 their modifications 546:564 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 2 19 theme T 359:359 arg1 population 366:375 this γδ T cell population 351:375 this γδ T cell population 351:375 However, the molecular mechanisms behind the activation of this γδ T cell population remains unclear. 22846996 4 20 theme Ig 958:959 arg1 domain 961:966 the membrane proximal C-type Ig domain 929:966 the membrane proximal C-type Ig domain 929:966 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 2 21 theme γδ 356:357 arg1 population 366:375 this γδ T cell population 351:375 this γδ T cell population 351:375 However, the molecular mechanisms behind the activation of this γδ T cell population remains unclear. 22846996 1 22 theme endogenous 248:257 arg1 phosphoisoprenoids 272:289 endogenous or exogenous phosphoisoprenoids 248:289 endogenous or exogenous phosphoisoprenoids 248:289 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 1 23 theme rapid 167:171 arg1 response 184:191 their rapid and potent response 161:191 their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids 161:289 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 0 24 theme -specific 99:107 arg1 antibodies 109:118 CD277/butyrophilin-3 (BTN3A)-specific antibodies 71:118 CD277/butyrophilin-3 (BTN3A)-specific antibodies 71:118 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 4 25 theme proximal 942:949 arg1 domain 961:966 the membrane proximal C-type Ig domain 929:966 the membrane proximal C-type Ig domain 929:966 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 1 26 theme exogenous 262:270 arg1 phosphoisoprenoids 272:289 endogenous or exogenous phosphoisoprenoids 248:289 endogenous or exogenous phosphoisoprenoids 248:289 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 5 27 theme separate 1007:1014 arg1 epitopes 1016:1023 separate epitopes 1007:1023 separate epitopes on the BTN3A Ig-V domain 1007:1048 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 1 28 theme potent 177:182 arg1 response 184:191 their rapid and potent response 161:191 their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids 161:289 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 4 29 theme membrane 933:940 arg1 domain 961:966 the membrane proximal C-type Ig domain 929:966 the membrane proximal C-type Ig domain 929:966 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 1 30 theme phosphoisoprenoids 272:289 arg1 presence 236:243 the presence 232:243 the presence of endogenous or exogenous phosphoisoprenoids 232:289 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 6 31 theme T 1371:1371 arg1 receptor 1378:1385 the Vγ9Vδ2 T cell receptor 1360:1385 the Vγ9Vδ2 T cell receptor 1360:1385 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 6 32 theme functional 1178:1187 arg1 studies 1189:1195 functional studies 1178:1195 functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation 1178:1274 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 0 33 theme Vγ9Vδ2 44:49 arg1 responses 58:66 human Vγ9Vδ2 T cell responses 38:66 human Vγ9Vδ2 T cell responses 38:66 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 5 34 from epitopes 1016:1023 arg1 domain 1043:1048 the BTN3A Ig-V domain 1028:1048 the BTN3A Ig-V domain 1028:1048 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 4 35 theme B7 836:837 arg1 superfamily 839:849 the B7 superfamily 832:849 the B7 superfamily of proteins 832:861 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 1 36 from response 184:191 arg1 presence 236:243 the presence 232:243 the presence of endogenous or exogenous phosphoisoprenoids 232:289 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 0 37 theme human 38:42 arg1 responses 58:66 human Vγ9Vδ2 T cell responses 38:66 human Vγ9Vδ2 T cell responses 38:66 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 5 38 theme different 1085:1093 arg1 valencies 1095:1103 different valencies 1085:1103 different valencies based on their binding orientation 1085:1138 The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. 22846996 6 39 theme Vγ9Vδ2 1258:1263 arg1 activation 1265:1274 Vγ9Vδ2 activation 1258:1274 Vγ9Vδ2 activation 1258:1274 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 3 40 theme Vγ9Vδ2 648:653 arg1 activation 655:664 phosphoisoprenoid-mediated Vγ9Vδ2 activation 621:664 phosphoisoprenoid-mediated Vγ9Vδ2 activation 621:664 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 6 41 theme system 1205:1210 arg1 studies 1189:1195 functional studies 1178:1195 functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation 1178:1274 These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor. 22846996 4 42 theme BTN3A 755:759 arg1 isoforms 761:768 the three BTN3A isoforms 745:768 the three BTN3A isoforms 745:768 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 42 theme BTN3A 755:759 arg1 BTN3A2 779:784 BTN3A2 779:784 BTN3A2 779:784 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 42 theme BTN3A 755:759 arg1 BTN3A3 791:796 BTN3A3 791:796 BTN3A3 791:796 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 42 theme BTN3A 755:759 arg1 BTN3A1 771:776 BTN3A1 771:776 BTN3A1 771:776 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 3 43 theme agonist 585:591 arg1 20.1 603:606 20.1 603:606 20.1 603:606 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 3 43 theme agonist 585:591 arg1 antibody 593:600 an agonist antibody 582:600 an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation 582:664 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 2 44 theme molecular 305:313 arg1 mechanisms 315:324 the molecular mechanisms 301:324 the molecular mechanisms behind the activation of this γδ T cell population 301:375 However, the molecular mechanisms behind the activation of this γδ T cell population remains unclear. 22846996 4 45 contain have 799:802 arg1 isoforms 761:768 the three BTN3A isoforms 745:768 the three BTN3A isoforms 745:768 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 45 contain have 799:802 arg2 homology 820:827 high structural homology 804:827 high structural homology to the B7 superfamily of proteins 804:861 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 45 contain have 799:802 arg1 BTN3A2 779:784 BTN3A2 779:784 BTN3A2 779:784 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 45 contain have 799:802 arg1 BTN3A3 791:796 BTN3A3 791:796 BTN3A3 791:796 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 45 contain have 799:802 arg1 BTN3A1 771:776 BTN3A1 771:776 BTN3A1 771:776 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 1 46 theme Human 121:125 arg1 cells 136:140 Human Vγ9Vδ2 T cells 121:140 Human Vγ9Vδ2 T cells 121:140 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 3 47 theme BTN3A 452:456 arg1 molecules 459:467 the CD277/butyrophilin-3 (BTN3A) molecules 426:467 the CD277/butyrophilin-3 (BTN3A) molecules 426:467 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 0 48 theme cell 53:56 arg1 responses 58:66 human Vγ9Vδ2 T cell responses 38:66 human Vγ9Vδ2 T cell responses 38:66 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 3 49 theme phosphoisoprenoid-mediated 621:646 arg1 activation 655:664 phosphoisoprenoid-mediated Vγ9Vδ2 activation 621:664 phosphoisoprenoid-mediated Vγ9Vδ2 activation 621:664 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 3 50 theme CD277/butyrophilin-3 430:449 arg1 molecules 459:467 the CD277/butyrophilin-3 (BTN3A) molecules 426:467 the CD277/butyrophilin-3 (BTN3A) molecules 426:467 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 1 51 theme Vγ9Vδ2 127:132 arg1 cells 136:140 Human Vγ9Vδ2 T cells 121:140 Human Vγ9Vδ2 T cells 121:140 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 22846996 4 52 dep isoforms 761:768 arg1 isoforms 761:768 the three BTN3A isoforms 745:768 the three BTN3A isoforms 745:768 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 52 dep isoforms 761:768 arg1 BTN3A2 779:784 BTN3A2 779:784 BTN3A2 779:784 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 52 dep isoforms 761:768 arg1 BTN3A3 791:796 BTN3A3 791:796 BTN3A3 791:796 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 4 52 dep isoforms 761:768 arg1 BTN3A1 771:776 BTN3A1 771:776 BTN3A1 771:776 We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. 22846996 0 53 theme T 51:51 arg1 responses 58:66 human Vγ9Vδ2 T cell responses 38:66 human Vγ9Vδ2 T cell responses 38:66 The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies. 22846996 3 54 theme antagonist 673:682 arg1 103.2 694:698 103.2 694:698 103.2 694:698 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 3 54 theme antagonist 673:682 arg1 antibody 684:691 an antagonist antibody 670:691 an antagonist antibody (103.2) that inhibits this reactivity 670:729 Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. 22846996 1 55 theme T 134:134 arg1 cells 136:140 Human Vγ9Vδ2 T cells 121:140 Human Vγ9Vδ2 T cells 121:140 Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. 26134396 1 0 theme model 270:274 arg1 systems 276:282 various model systems 262:282 various model systems 262:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 1 1 from toxicity 250:257 arg1 systems 276:282 various model systems 262:282 various model systems 262:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 5 2 theme P2 988:989 arg1 availability 958:969 the availability 954:969 the availability of PA and PI(3,5)P2 954:989 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 6 3 from stress 1209:1214 arg1 disorders 1233:1241 related disorders 1225:1241 related disorders 1225:1241 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 6 3 from stress 1209:1214 arg1 PD 1219:1220 PD 1219:1220 PD 1219:1220 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 1 4 theme Mn 231:232 arg1 toxicity 250:257 α-synuclein, Mn(2+), and Zn(2+) toxicity 218:257 α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems 218:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 3 5 theme [PI 630:632 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 5 theme [PI 630:632 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 6 theme membrane 476:483 arg1 surface 485:491 the cytosolic membrane surface 462:491 the cytosolic membrane surface of the lysosome 462:507 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 7 theme lysosome 500:507 arg1 surface 485:491 the cytosolic membrane surface 462:491 the cytosolic membrane surface of the lysosome 462:507 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 5 8 theme active 849:854 arg1 ATP13A2 856:862 only catalytically active ATP13A2 830:862 only catalytically active ATP13A2 830:862 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 6 9 theme α-synuclein 1171:1181 arg1 toxicity 1183:1190 α-synuclein toxicity 1171:1190 α-synuclein toxicity 1171:1190 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 5 10 theme 3,5 984:986 arg1 P2 988:989 PI(3,5)P2 981:989 PI(3,5)P2 981:989 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 4 11 theme PI 747:748 arg1 P2 754:755 PI(3,5)P2 747:755 PI(3,5)P2 747:755 We further demonstrate that ATP13A2 accumulates in an inactive autophosphorylated state and that PA and PI(3,5)P2 stimulate the autophosphorylation of ATP13A2. 26134396 6 12 theme PI 1031:1032 arg1 P2 1038:1039 PI(3,5)P2 1031:1039 PI(3,5)P2 1031:1039 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 3 13 theme cytosolic 466:474 arg1 surface 485:491 the cytosolic membrane surface 462:491 the cytosolic membrane surface of the lysosome 462:507 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 0 14 theme lipid 2:6 arg1 switch 8:13 A lipid switch 0:13 A lipid switch 0:13 A lipid switch unlocks Parkinson's disease-associated ATP13A2. 26134396 3 15 theme 3,5 634:636 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 15 theme 3,5 634:636 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 4 16 theme 3,5 750:752 arg1 P2 754:755 PI(3,5)P2 747:755 PI(3,5)P2 747:755 We further demonstrate that ATP13A2 accumulates in an inactive autophosphorylated state and that PA and PI(3,5)P2 stimulate the autophosphorylation of ATP13A2. 26134396 1 17 theme lysosomal 76:84 arg1 ATP13A2 63:69 ATP13A2 63:69 ATP13A2 63:69 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 1 17 theme lysosomal 76:84 arg1 ATPase 103:108 a lysosomal P-type transport ATPase 74:108 a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems 74:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 6 18 theme ATP13A2 1084:1090 arg1 activity 1072:1079 the activity 1068:1079 the activity 1068:1079 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 1 19 theme P-type 86:91 arg1 ATP13A2 63:69 ATP13A2 63:69 ATP13A2 63:69 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 1 19 theme P-type 86:91 arg1 ATPase 103:108 a lysosomal P-type transport ATPase 74:108 a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems 74:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 2 20 theme molecular 297:305 arg1 function 307:314 molecular function 297:314 molecular function 297:314 So far, the molecular function and regulation of ATP13A2 remains undetermined. 26134396 6 21 theme P2 1038:1039 arg1 binding 1013:1019 the N-terminal binding 998:1019 the N-terminal binding of PA and PI(3,5)P2 998:1039 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 1 22 theme transport 93:101 arg1 ATP13A2 63:69 ATP13A2 63:69 ATP13A2 63:69 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 1 22 theme transport 93:101 arg1 ATPase 103:108 a lysosomal P-type transport ATPase 74:108 a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems 74:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 6 23 from toxicity 1183:1190 arg1 disorders 1233:1241 related disorders 1225:1241 related disorders 1225:1241 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 6 23 from toxicity 1183:1190 arg1 PD 1219:1220 PD 1219:1220 PD 1219:1220 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 5 24 theme PD 826:827 arg1 model 817:821 a cellular model 806:821 a cellular model of PD 806:827 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 1 25 theme Zn 243:244 arg1 toxicity 250:257 α-synuclein, Mn(2+), and Zn(2+) toxicity 218:257 α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems 218:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 6 26 theme N-terminal 1002:1011 arg1 binding 1013:1019 the N-terminal binding 998:1019 the N-terminal binding of PA and PI(3,5)P2 998:1039 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 3 27 theme phosphatidic 565:576 arg1 PA 584:585 PA 584:585 PA 584:585 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 27 theme phosphatidic 565:576 arg1 acid 578:581 phosphatidic acid 565:581 phosphatidic acid (PA) 565:586 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 27 theme phosphatidic 565:576 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 5 28 theme rotenone-induced 899:914 arg1 stress 930:935 rotenone-induced mitochondrial stress 899:935 rotenone-induced mitochondrial stress 899:935 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 5 29 theme PA 974:975 arg1 availability 958:969 the availability 954:969 the availability of PA and PI(3,5)P2 954:989 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 5 30 theme mitochondrial 916:928 arg1 stress 930:935 rotenone-induced mitochondrial stress 899:935 rotenone-induced mitochondrial stress 899:935 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 2 31 theme ATP13A2 334:340 arg1 regulation 320:329 regulation 320:329 regulation 320:329 So far, the molecular function and regulation of ATP13A2 remains undetermined. 26134396 2 31 theme ATP13A2 334:340 arg1 function 307:314 molecular function 297:314 molecular function 297:314 So far, the molecular function and regulation of ATP13A2 remains undetermined. 26134396 6 32 dep activate 1135:1142 arg1 reduce 1164:1169 reduce 1164:1169 reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders 1164:1241 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 3 33 contain contains 398:405 arg1 ATP13A2 390:396 ATP13A2 390:396 ATP13A2 390:396 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 33 contain contains 398:405 arg2 extension 439:447 a unique N-terminal hydrophobic extension 407:447 a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 407:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 34 dep lipids 558:563 arg1 PA 584:585 PA 584:585 PA 584:585 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 34 dep lipids 558:563 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 34 dep lipids 558:563 arg1 acid 578:581 phosphatidic acid 565:581 phosphatidic acid (PA) 565:586 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 34 dep lipids 558:563 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 6 35 theme related 1225:1231 arg1 disorders 1233:1241 related disorders 1225:1241 related disorders 1225:1241 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 5 36 theme PI 981:982 arg1 P2 988:989 PI(3,5)P2 981:989 PI(3,5)P2 981:989 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 3 37 theme N-terminal 416:425 arg1 extension 439:447 a unique N-terminal hydrophobic extension 407:447 a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 407:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 0 38 theme disease-associated 35:52 arg1 ATP13A2 54:60 Parkinson's disease-associated ATP13A2 23:60 Parkinson's disease-associated ATP13A2 23:60 A lipid switch unlocks Parkinson's disease-associated ATP13A2. 26134396 1 39 theme various 262:268 arg1 systems 276:282 various model systems 262:282 various model systems 262:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 3 40 theme phosphatidylinositol 592:611 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 40 theme phosphatidylinositol 592:611 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 41 theme hydrophobic 427:437 arg1 extension 439:447 a unique N-terminal hydrophobic extension 407:447 a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 407:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 4 42 theme inactive 697:704 arg1 state 725:729 an inactive autophosphorylated state 694:729 an inactive autophosphorylated state 694:729 We further demonstrate that ATP13A2 accumulates in an inactive autophosphorylated state and that PA and PI(3,5)P2 stimulate the autophosphorylation of ATP13A2. 26134396 6 43 theme therapeutic 1111:1121 arg1 strategy 1123:1130 a therapeutic strategy 1109:1130 a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders 1109:1241 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 3 44 theme signaling 548:556 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 44 theme signaling 548:556 arg1 acid 578:581 phosphatidic acid 565:581 phosphatidic acid (PA) 565:586 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 44 theme signaling 548:556 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 2 45 dep function 307:314 arg1 the 293:295 the 293:295 the 293:295 So far, the molecular function and regulation of ATP13A2 remains undetermined. 26134396 5 46 theme cellular 808:815 arg1 model 817:821 a cellular model 806:821 a cellular model of PD 806:827 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 4 47 theme ATP13A2 794:800 arg1 autophosphorylation 771:789 the autophosphorylation 767:789 the autophosphorylation of ATP13A2 767:800 We further demonstrate that ATP13A2 accumulates in an inactive autophosphorylated state and that PA and PI(3,5)P2 stimulate the autophosphorylation of ATP13A2. 26134396 6 48 theme 3,5 1034:1036 arg1 P2 1038:1039 PI(3,5)P2 1031:1039 PI(3,5)P2 1031:1039 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 3 49 theme unique 409:414 arg1 extension 439:447 a unique N-terminal hydrophobic extension 407:447 a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 407:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 6 50 theme mitochondrial 1195:1207 arg1 stress 1209:1214 mitochondrial stress 1195:1214 mitochondrial stress 1195:1214 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 4 51 theme autophosphorylated 706:723 arg1 state 725:729 an inactive autophosphorylated state 694:729 an inactive autophosphorylated state 694:729 We further demonstrate that ATP13A2 accumulates in an inactive autophosphorylated state and that PA and PI(3,5)P2 stimulate the autophosphorylation of ATP13A2. 26134396 6 52 theme PA 1024:1025 arg1 binding 1013:1019 the N-terminal binding 998:1019 the N-terminal binding of PA and PI(3,5)P2 998:1039 Thus, the N-terminal binding of PA and PI(3,5)P2 emerges as a key to unlock the activity of ATP13A2, which may offer a therapeutic strategy to activate ATP13A2 and thereby reduce α-synuclein toxicity or mitochondrial stress in PD or related disorders. 26134396 1 53 theme α-synuclein 218:228 arg1 toxicity 250:257 α-synuclein, Mn(2+), and Zn(2+) toxicity 218:257 α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems 218:282 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 26134396 5 54 theme cellular 871:878 arg1 protection 880:889 cellular protection 871:889 cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2 871:989 In a cellular model of PD, only catalytically active ATP13A2 offers cellular protection against rotenone-induced mitochondrial stress, which relies on the availability of PA and PI(3,5)P2. 26134396 3 55 theme bisphosphate 617:628 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 55 theme bisphosphate 617:628 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 56 theme lysosomal 538:546 arg1 P2 638:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2 592:639 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 56 theme lysosomal 538:546 arg1 acid 578:581 phosphatidic acid 565:581 phosphatidic acid (PA) 565:586 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 3 56 theme lysosomal 538:546 arg1 lipids 558:563 the lysosomal signaling lipids 534:563 the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2] 534:640 Here, we demonstrate that ATP13A2 contains a unique N-terminal hydrophobic extension that lies on the cytosolic membrane surface of the lysosome, where it interacts with the lysosomal signaling lipids phosphatidic acid (PA) and phosphatidylinositol(3,5)bisphosphate [PI(3,5)P2]. 26134396 1 57 theme Kufor-Rakeb 138:148 arg1 syndrome 150:157 Kufor-Rakeb syndrome 138:157 Kufor-Rakeb syndrome 138:157 ATP13A2 is a lysosomal P-type transport ATPase that has been implicated in Kufor-Rakeb syndrome and Parkinson's disease (PD), providing protection against α-synuclein, Mn(2+), and Zn(2+) toxicity in various model systems. 3198637 0 0 theme type 61:64 arg1 collagen 69:76 human type IV collagen 55:76 human type IV collagen 55:76 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 1 1 theme clones 253:258 arg1 sequencing 234:243 nucleotide sequencing 223:243 nucleotide sequencing of cDNA clones 223:258 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 4 2 theme translated 613:622 arg1 polypeptide 624:634 The translated polypeptide 609:634 The translated polypeptide 609:634 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 peptide 649:655 a signal peptide 640:655 a signal peptide of 36 amino acids 640:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 part 709:712 an amino-terminal noncollagenous part 676:712 an amino-terminal noncollagenous part of 21 residues 676:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 domain 758:763 a 1,428-residue collagenous domain 730:763 a 1,428-residue collagenous domain with 23 interruptions 730:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg2 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 3 contain has 636:638 arg1 polypeptide 624:634 The translated polypeptide 609:634 The translated polypeptide 609:634 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 2 4 theme cDNA 277:280 arg1 clones 282:287 The overlapping cDNA clones 261:287 The overlapping cDNA clones 261:287 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 4 5 theme noncollagenous 812:825 arg1 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 5 theme noncollagenous 812:825 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 5 theme noncollagenous 812:825 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 5 theme noncollagenous 812:825 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 0 6 theme human 55:59 arg1 collagen 69:76 human type IV collagen 55:76 human type IV collagen 55:76 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 2 7 theme overlapping 265:275 arg1 clones 282:287 The overlapping cDNA clones 261:287 The overlapping cDNA clones 261:287 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 4 8 with domain 758:763 arg1 interruptions 773:785 23 interruptions 770:785 23 interruptions 770:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 0 9 theme collagen 69:76 arg1 chain 46:50 the alpha 2 chain 34:50 the alpha 2 chain of human type IV collagen 34:76 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 0 9 theme collagen 69:76 arg1 comparison 82:91 comparison 82:91 comparison with the alpha 1(IV) chain 82:118 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 2 10 with pairs 306:310 arg1 region 425:430 the 3'-untranslated region 405:430 the 3'-untranslated region of 838 base pairs 405:448 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 10 with pairs 306:310 arg1 region 335:340 a 5'-untranslated region 317:340 a 5'-untranslated region of 283 base pairs 317:358 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 10 with pairs 306:310 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 5 11 theme molecular 871:879 arg1 Da 935:936 163,774 Da 927:936 163,774 Da 927:936 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 5 11 theme molecular 871:879 arg1 mass 881:884 The calculated molecular mass 856:884 The calculated molecular mass of the mature human alpha 2(IV) chain 856:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 5 12 theme calculated 860:869 arg1 Da 935:936 163,774 Da 927:936 163,774 Da 927:936 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 5 12 theme calculated 860:869 arg1 mass 881:884 The calculated molecular mass 856:884 The calculated molecular mass of the mature human alpha 2(IV) chain 856:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 1 13 theme IV 190:191 arg1 chain 194:198 the human type IV collagen alpha 2(IV) chain 155:198 the human type IV collagen alpha 2(IV) chain 155:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 4 14 theme residues 846:853 arg1 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 14 theme residues 846:853 arg1 domain 758:763 a 1,428-residue collagenous domain 730:763 a 1,428-residue collagenous domain with 23 interruptions 730:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 14 theme residues 846:853 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 14 theme residues 846:853 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 14 theme residues 846:853 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 14 theme residues 846:853 arg1 peptide 649:655 a signal peptide 640:655 a signal peptide of 36 amino acids 640:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 14 theme residues 846:853 arg1 part 709:712 an amino-terminal noncollagenous part 676:712 an amino-terminal noncollagenous part of 21 residues 676:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 15 with domain 832:837 arg1 interruptions 773:785 23 interruptions 770:785 23 interruptions 770:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 domain 758:763 a 1,428-residue collagenous domain 730:763 a 1,428-residue collagenous domain with 23 interruptions 730:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 peptide 649:655 a signal peptide 640:655 a signal peptide of 36 amino acids 640:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 16 theme acids 669:673 arg1 part 709:712 an amino-terminal noncollagenous part 676:712 an amino-terminal noncollagenous part of 21 residues 676:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 0 17 theme comparison 82:91 arg1 structure 21:29 The complete primary structure 0:29 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 0:119 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 5 18 theme human 900:904 arg1 chain 918:922 the mature human alpha 2(IV) chain 889:922 the mature human alpha 2(IV) chain 889:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 0 19 with chain 46:50 arg1 chain 114:118 the alpha 1(IV) chain 98:118 the alpha 1(IV) chain 98:118 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 4 20 theme amino-terminal 679:692 arg1 part 709:712 an amino-terminal noncollagenous part 676:712 an amino-terminal noncollagenous part of 21 residues 676:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 2 21 theme base 301:304 arg1 pairs 306:310 6,257 base pairs 295:310 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs 295:448 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 4 22 theme collagenous 746:756 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 22 theme collagenous 746:756 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 22 theme collagenous 746:756 arg1 domain 758:763 a 1,428-residue collagenous domain 730:763 a 1,428-residue collagenous domain with 23 interruptions 730:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 22 theme collagenous 746:756 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 0 23 with comparison 82:91 arg1 chain 114:118 the alpha 1(IV) chain 98:118 the alpha 1(IV) chain 98:118 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 4 24 theme 227 842:844 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 2 25 theme 3'-untranslated 409:423 arg1 region 425:430 the 3'-untranslated region 405:430 the 3'-untranslated region of 838 base pairs 405:448 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 0 26 theme complete 4:11 arg1 structure 21:29 The complete primary structure 0:29 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 0:119 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 2 27 theme reading 386:392 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 3 28 theme amino 465:469 arg1 sequence 476:483 The predicted amino acid sequence 451:483 The predicted amino acid sequence 451:483 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 1 29 theme complete 125:132 arg1 structure 142:150 The complete primary structure 121:150 The complete primary structure of the human type IV collagen alpha 2(IV) chain 121:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 2 30 theme pairs 354:358 arg1 region 425:430 the 3'-untranslated region 405:430 the 3'-untranslated region of 838 base pairs 405:448 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 30 theme pairs 354:358 arg1 region 335:340 a 5'-untranslated region 317:340 a 5'-untranslated region of 283 base pairs 317:358 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 30 theme pairs 354:358 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 1 31 theme chain 194:198 arg1 structure 142:150 The complete primary structure 121:150 The complete primary structure of the human type IV collagen alpha 2(IV) chain 121:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 3 32 theme acid 471:474 arg1 sequence 476:483 The predicted amino acid sequence 451:483 The predicted amino acid sequence 451:483 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 1 33 theme primary 134:140 arg1 structure 142:150 The complete primary structure 121:150 The complete primary structure of the human type IV collagen alpha 2(IV) chain 121:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 2 34 theme base 349:352 arg1 pairs 354:358 283 base pairs 345:358 283 base pairs 345:358 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 5 35 theme alpha 906:910 arg1 chain 918:922 the mature human alpha 2(IV) chain 889:922 the mature human alpha 2(IV) chain 889:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 4 36 theme amino 663:667 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 0 37 theme primary 13:19 arg1 structure 21:29 The complete primary structure 0:29 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 0:119 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 2 38 theme pairs 444:448 arg1 region 425:430 the 3'-untranslated region 405:430 the 3'-untranslated region of 838 base pairs 405:448 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 38 theme pairs 444:448 arg1 region 335:340 a 5'-untranslated region 317:340 a 5'-untranslated region of 283 base pairs 317:358 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 38 theme pairs 444:448 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 3 39 theme predicted 455:463 arg1 sequence 476:483 The predicted amino acid sequence 451:483 The predicted amino acid sequence 451:483 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 2 40 theme open 381:384 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 41 theme base 439:442 arg1 pairs 444:448 838 base pairs 435:448 838 base pairs 435:448 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 42 theme 5'-untranslated 319:333 arg1 region 335:340 a 5'-untranslated region 317:340 a 5'-untranslated region of 283 base pairs 317:358 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 2 43 theme pair 376:379 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 0 44 theme alpha 102:106 arg1 chain 114:118 the alpha 1(IV) chain 98:118 the alpha 1(IV) chain 98:118 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 4 45 theme residues 720:727 arg1 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 45 theme residues 720:727 arg1 domain 758:763 a 1,428-residue collagenous domain 730:763 a 1,428-residue collagenous domain with 23 interruptions 730:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 45 theme residues 720:727 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 45 theme residues 720:727 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 45 theme residues 720:727 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 45 theme residues 720:727 arg1 peptide 649:655 a signal peptide 640:655 a signal peptide of 36 amino acids 640:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 45 theme residues 720:727 arg1 part 709:712 an amino-terminal noncollagenous part 676:712 an amino-terminal noncollagenous part of 21 residues 676:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 1 46 theme human 159:163 arg1 chain 194:198 the human type IV collagen alpha 2(IV) chain 155:198 the human type IV collagen alpha 2(IV) chain 155:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 2 47 theme 5,136-base 365:374 arg1 frame 394:398 the 5,136-base pair open reading frame 361:398 the 5,136-base pair open reading frame 361:398 The overlapping cDNA clones cover 6,257 base pairs with a 5'-untranslated region of 283 base pairs, the 5,136-base pair open reading frame, and the 3'-untranslated region of 838 base pairs. 3198637 1 48 theme nucleotide 223:232 arg1 sequencing 234:243 nucleotide sequencing 223:243 nucleotide sequencing of cDNA clones 223:258 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 0 49 theme IV 110:111 arg1 chain 114:118 the alpha 1(IV) chain 98:118 the alpha 1(IV) chain 98:118 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 3 50 theme 1,712 548:552 arg1 residues 554:561 1,712 residues 548:561 1,712 residues corresponding in molecular weight to 167,560 548:606 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 4 51 theme 1,428-residue 732:744 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 51 theme 1,428-residue 732:744 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 51 theme 1,428-residue 732:744 arg1 domain 758:763 a 1,428-residue collagenous domain 730:763 a 1,428-residue collagenous domain with 23 interruptions 730:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 51 theme 1,428-residue 732:744 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 0 52 theme alpha 38:42 arg1 chain 46:50 the alpha 2 chain 34:50 the alpha 2 chain of human type IV collagen 34:76 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 4 53 theme carboxyl-terminal 794:810 arg1 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 53 theme carboxyl-terminal 794:810 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 53 theme carboxyl-terminal 794:810 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 53 theme carboxyl-terminal 794:810 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 5 54 theme chain 918:922 arg1 Da 935:936 163,774 Da 927:936 163,774 Da 927:936 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 5 54 theme chain 918:922 arg1 mass 881:884 The calculated molecular mass 856:884 The calculated molecular mass of the mature human alpha 2(IV) chain 856:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 4 55 with peptide 649:655 arg1 interruptions 773:785 23 interruptions 770:785 23 interruptions 770:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 56 theme NC 828:829 arg1 domain 832:837 a carboxyl-terminal noncollagenous (NC) domain 792:837 a carboxyl-terminal noncollagenous (NC) domain of 227 residues 792:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 56 theme NC 828:829 arg1 residues 846:853 227 residues 842:853 227 residues 842:853 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 56 theme NC 828:829 arg1 acids 669:673 36 amino acids 660:673 36 amino acids 660:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 4 56 theme NC 828:829 arg1 residues 720:727 21 residues 717:727 21 residues 717:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 5 57 theme mature 893:898 arg1 chain 918:922 the mature human alpha 2(IV) chain 889:922 the mature human alpha 2(IV) chain 889:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 5 58 theme IV 914:915 arg1 chain 918:922 the mature human alpha 2(IV) chain 889:922 the mature human alpha 2(IV) chain 889:922 The calculated molecular mass of the mature human alpha 2(IV) chain is 163,774 Da. 3198637 4 59 theme signal 642:647 arg1 peptide 649:655 a signal peptide 640:655 a signal peptide of 36 amino acids 640:673 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 1 60 theme type 165:168 arg1 chain 194:198 the human type IV collagen alpha 2(IV) chain 155:198 the human type IV collagen alpha 2(IV) chain 155:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 4 61 theme noncollagenous 694:707 arg1 part 709:712 an amino-terminal noncollagenous part 676:712 an amino-terminal noncollagenous part of 21 residues 676:727 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 1 62 theme IV 170:171 arg1 chain 194:198 the human type IV collagen alpha 2(IV) chain 155:198 the human type IV collagen alpha 2(IV) chain 155:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 3 63 theme molecular 580:588 arg1 weight 590:595 molecular weight 580:595 molecular weight to 167,560 580:606 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 3 64 theme complete 507:514 arg1 product 528:534 the complete translation product 503:534 the complete translation product 503:534 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 1 65 theme collagen 173:180 arg1 chain 194:198 the human type IV collagen alpha 2(IV) chain 155:198 the human type IV collagen alpha 2(IV) chain 155:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 0 66 theme chain 46:50 arg1 structure 21:29 The complete primary structure 0:29 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 0:119 The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. 3198637 1 67 theme cDNA 248:251 arg1 clones 253:258 cDNA clones 248:258 cDNA clones 248:258 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 3198637 3 68 theme translation 516:526 arg1 product 528:534 the complete translation product 503:534 the complete translation product 503:534 The predicted amino acid sequence demonstrates that the complete translation product consists of 1,712 residues corresponding in molecular weight to 167,560. 3198637 4 69 with part 709:712 arg1 interruptions 773:785 23 interruptions 770:785 23 interruptions 770:785 The translated polypeptide has a signal peptide of 36 amino acids, an amino-terminal noncollagenous part of 21 residues, a 1,428-residue collagenous domain with 23 interruptions, and a carboxyl-terminal noncollagenous (NC) domain of 227 residues. 3198637 1 70 theme alpha 182:186 arg1 chain 194:198 the human type IV collagen alpha 2(IV) chain 155:198 the human type IV collagen alpha 2(IV) chain 155:198 The complete primary structure of the human type IV collagen alpha 2(IV) chain has been determined by nucleotide sequencing of cDNA clones. 2493678 3 0 from angstroms 379:387 arg1 data 358:361 data 358:361 data from 8.0 to 2.5 angstroms (A) 358:391 The present agreement factor for data from 8.0 to 2.5 angstroms (A) is 0.236. 2493678 1 1 theme recombinant 132:142 arg1 renin 150:154 recombinant human renin 132:154 recombinant human renin 132:154 The x-ray crystal structure of recombinant human renin has been determined. 2493678 4 2 theme disordered 462:471 arg1 regions 473:479 these disordered regions 456:479 these disordered regions 456:479 Some of the surface loops are poorly determined, and these disordered regions border a 30 A wide solvent channel. 2493678 5 3 theme structural 594:603 arg1 cores 605:609 the structural cores 590:609 the structural cores 590:609 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 1 4 theme human 144:148 arg1 renin 150:154 recombinant human renin 132:154 recombinant human renin 132:154 The x-ray crystal structure of recombinant human renin has been determined. 2493678 5 5 theme other 542:546 arg1 proteinases 557:567 other aspartyl proteinases 542:567 other aspartyl proteinases 542:567 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 6 6 theme inhibitors 869:878 arg1 design 853:858 the design 849:858 the design of renin inhibitors 849:878 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 6 7 theme renin 863:867 arg1 inhibitors 869:878 renin inhibitors 863:878 renin inhibitors 863:878 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 5 8 theme aspartyl 548:555 arg1 proteinases 557:567 other aspartyl proteinases 542:567 other aspartyl proteinases 542:567 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 2 9 theme porcine 305:311 arg1 pepsinogen 313:322 porcine pepsinogen 305:322 porcine pepsinogen 305:322 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 2493678 3 10 theme present 329:335 arg1 0.236 396:400 0.236 396:400 0.236 396:400 The present agreement factor for data from 8.0 to 2.5 angstroms (A) is 0.236. 2493678 3 10 theme present 329:335 arg1 factor 347:352 The present agreement factor 325:352 The present agreement factor for data from 8.0 to 2.5 angstroms (A) 325:391 The present agreement factor for data from 8.0 to 2.5 angstroms (A) is 0.236. 2493678 3 11 dep 2.5 375:377 arg1 to 372:373 to 372:373 to 372:373 The present agreement factor for data from 8.0 to 2.5 angstroms (A) is 0.236. 2493678 1 12 theme renin 150:154 arg1 structure 119:127 The x-ray crystal structure 101:127 The x-ray crystal structure of recombinant human renin 101:154 The x-ray crystal structure of recombinant human renin has been determined. 2493678 0 13 theme A 87:87 arg1 resolution 89:98 2.5 A resolution 83:98 2.5 A resolution 83:98 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 5 14 with Comparison 517:526 arg1 proteinases 557:567 other aspartyl proteinases 542:567 other aspartyl proteinases 542:567 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 6 15 theme models 798:803 arg1 use 791:793 the use 787:793 the use of models based on related enzymes 787:828 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 4 16 theme wide 495:498 arg1 channel 508:514 A wide solvent channel 493:514 a 30 A wide solvent channel 488:514 Some of the surface loops are poorly determined, and these disordered regions border a 30 A wide solvent channel. 2493678 5 17 dep vary 712:715 arg1 10A 732:734 10A 732:734 10A 732:734 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 0 18 theme human 25:29 arg1 renin 31:35 recombinant human renin 13:35 recombinant human renin 13:35 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 0 18 theme human 25:29 arg1 target 40:45 a target 38:45 a target for cardiovascular-active drugs 38:77 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 5 19 theme renin 531:535 arg1 Comparison 517:526 Comparison 517:526 Comparison of renin with other aspartyl proteinases 517:567 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 0 20 theme recombinant 13:23 arg1 renin 31:35 recombinant human renin 13:35 recombinant human renin 13:35 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 0 20 theme recombinant 13:23 arg1 target 40:45 a target 38:45 a target for cardiovascular-active drugs 38:77 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 2 21 theme initial 282:288 arg1 model 290:294 an initial model 279:294 an initial model based on porcine pepsinogen 279:322 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 2493678 4 22 theme solvent 500:506 arg1 channel 508:514 A wide solvent channel 493:514 a 30 A wide solvent channel 488:514 Some of the surface loops are poorly determined, and these disordered regions border a 30 A wide solvent channel. 2493678 5 23 theme surface 650:656 arg1 residues 658:665 surface residues 650:665 surface residues 650:665 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 0 24 theme renin 31:35 arg1 Structure 0:8 Structure 0:8 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 0:99 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 6 25 dep structure 762:770 arg1 opposed 776:782 opposed 776:782 opposed to the use of models based on related enzymes 776:828 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 4 26 theme surface 415:421 arg1 loops 423:427 the surface loops 411:427 the surface loops 411:427 Some of the surface loops are poorly determined, and these disordered regions border a 30 A wide solvent channel. 2493678 2 27 theme crystallographic 221:236 arg1 data 238:241 crystallographic data 221:241 crystallographic data 221:241 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 2493678 2 27 theme crystallographic 221:236 arg1 restraint 248:256 a restraint 246:256 a restraint 246:256 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 2493678 3 28 theme agreement 337:345 arg1 0.236 396:400 0.236 396:400 0.236 396:400 The present agreement factor for data from 8.0 to 2.5 angstroms (A) is 0.236. 2493678 3 28 theme agreement 337:345 arg1 factor 347:352 The present agreement factor 325:352 The present agreement factor for data from 8.0 to 2.5 angstroms (A) 325:391 The present agreement factor for data from 8.0 to 2.5 angstroms (A) is 0.236. 2493678 5 29 theme active 615:620 arg1 sites 622:626 active sites 615:626 active sites 615:626 Comparison of renin with other aspartyl proteinases shows that, although the structural cores and active sites are highly conserved, surface residues, some of which are critical for specificity, vary greatly (up to 10A). 2493678 6 30 theme actual 755:760 arg1 structure 762:770 the actual structure 751:770 the actual structure 751:770 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 0 31 from resolution 89:98 arg1 Structure 0:8 Structure 0:8 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 0:99 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 6 32 theme structure 762:770 arg1 Knowledge 738:746 Knowledge 738:746 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, 738:829 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 6 33 theme related 814:820 arg1 enzymes 822:828 related enzymes 814:828 related enzymes 814:828 Knowledge of the actual structure, as opposed to the use of models based on related enzymes, should facilitate the design of renin inhibitors. 2493678 2 34 theme dynamics 187:194 arg1 techniques 196:205 Molecular dynamics techniques 177:205 Molecular dynamics techniques that included crystallographic data as a restraint 177:256 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 2493678 1 35 theme x-ray 105:109 arg1 structure 119:127 The x-ray crystal structure 101:127 The x-ray crystal structure of recombinant human renin 101:154 The x-ray crystal structure of recombinant human renin has been determined. 2493678 2 36 theme Molecular 177:185 arg1 techniques 196:205 Molecular dynamics techniques 177:205 Molecular dynamics techniques that included crystallographic data as a restraint 177:256 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 2493678 1 37 theme crystal 111:117 arg1 structure 119:127 The x-ray crystal structure 101:127 The x-ray crystal structure of recombinant human renin 101:154 The x-ray crystal structure of recombinant human renin has been determined. 2493678 0 38 theme cardiovascular-active 51:71 arg1 drugs 73:77 cardiovascular-active drugs 51:77 cardiovascular-active drugs 51:77 Structure of recombinant human renin, a target for cardiovascular-active drugs, at 2.5 A resolution. 2493678 2 39 used used 263:266 arg2 techniques 196:205 Molecular dynamics techniques 177:205 Molecular dynamics techniques that included crystallographic data as a restraint 177:256 Molecular dynamics techniques that included crystallographic data as a restraint were used to improve an initial model based on porcine pepsinogen. 12775711 0 0 theme articular 89:97 arg1 chondrocytes 99:110 human articular chondrocytes 83:110 human articular chondrocytes 83:110 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 8 1 theme large 1123:1127 arg1 change 1144:1149 a large conformational change 1121:1149 a large conformational change 1121:1149 However, unlike the chitinases, binding of the oligosaccharide ligand to HCGP39 induces a large conformational change. 12775711 8 2 theme ligand 1096:1101 arg1 binding 1065:1071 binding 1065:1071 binding of the oligosaccharide ligand to HCGP39 1065:1111 However, unlike the chitinases, binding of the oligosaccharide ligand to HCGP39 induces a large conformational change. 12775711 5 3 theme Chi-lectins 738:748 arg1 ligands 723:729 any physiological or pathological ligands 689:729 any physiological or pathological ligands of the Chi-lectins 689:748 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 0 4 theme human 83:87 arg1 chondrocytes 99:110 human articular chondrocytes 83:110 human articular chondrocytes 83:110 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 4 5 theme HCGP39 507:512 arg1 ability 496:502 The ability 492:502 The ability of HCGP39 to activate cytoplasmic signaling pathways 492:555 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 5 6 theme interaction 786:796 arg1 identity 677:684 the identity 673:684 the identity of any physiological or pathological ligands of the Chi-lectins 673:748 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 5 6 theme interaction 786:796 arg1 nature 757:762 the nature 753:762 the nature of the protein-ligand interaction 753:796 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 5 7 theme ligands 723:729 arg1 identity 677:684 the identity 673:684 the identity of any physiological or pathological ligands of the Chi-lectins 673:748 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 5 7 theme ligands 723:729 arg1 nature 757:762 the nature 753:762 the nature of the protein-ligand interaction 753:796 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 0 8 gly glycoprotein 65:76 arg1 glycoprotein 65:76 the 39-kDa glycoprotein 54:76 the 39-kDa glycoprotein 54:76 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 8 9 theme conformational 1129:1142 arg1 change 1144:1149 a large conformational change 1121:1149 a large conformational change 1121:1149 However, unlike the chitinases, binding of the oligosaccharide ligand to HCGP39 induces a large conformational change. 12775711 7 10 with structures 897:906 arg1 GlcNAc8 949:955 GlcNAc8 949:955 GlcNAc8 949:955 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 1 11 theme articular 250:258 arg1 chondrocytes 260:271 articular chondrocytes 250:271 articular chondrocytes 250:271 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 3 12 theme cells 485:489 arg1 growth 457:462 the growth 453:462 the growth of connective tissue cells 453:489 Similar to other Chi-lectins, HCGP39 promotes the growth of connective tissue cells. 12775711 7 13 theme Crystal 889:895 arg1 structures 897:906 Crystal structures 889:906 Crystal structures of the native protein 889:928 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 4 14 theme ligand 584:589 arg1 presence 570:577 the presence 566:577 the presence of a ligand for this protein at the cell surface 566:626 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 4 15 attach presence 570:577 arg1 surface 620:626 the cell surface 611:626 the cell surface 611:626 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 4 15 attach presence 570:577 arg2 ligand 584:589 a ligand 582:589 a ligand for this protein 582:606 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 6 16 theme micromolar 868:877 arg1 affinity 879:886 micromolar affinity 868:886 micromolar affinity 868:886 Here, we show that HCGP39 is able to bind chitooligosaccharides with micromolar affinity. 12775711 2 17 from role 339:342 arg1 remodeling 365:374 connective tissue remodeling 347:374 connective tissue remodeling 347:374 Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. 12775711 9 18 theme chitinous 1279:1287 arg1 pathogens 1289:1297 chitinous pathogens 1279:1297 chitinous pathogens 1279:1297 Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. 12775711 9 18 theme chitinous 1279:1287 arg1 nematodes 1318:1326 nematodes 1318:1326 nematodes 1318:1326 Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. 12775711 9 18 theme chitinous 1279:1287 arg1 fungi 1308:1312 fungi 1308:1312 fungi 1308:1312 Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. 12775711 3 19 theme other 418:422 arg1 Chi-lectins 424:434 other Chi-lectins 418:434 other Chi-lectins 418:434 Similar to other Chi-lectins, HCGP39 promotes the growth of connective tissue cells. 12775711 7 20 theme protein 922:928 arg1 complex 936:942 a complex 934:942 a complex with GlcNAc8 934:955 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 7 20 theme protein 922:928 arg1 structures 897:906 Crystal structures 889:906 Crystal structures of the native protein 889:928 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 1 21 theme novel 177:181 arg1 family 183:188 a novel family 175:188 a novel family of chitinase-like lectins (Chilectins) 175:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 0 22 theme conformational 29:42 arg1 change 44:49 ligand-induced conformational change 14:49 ligand-induced conformational change 14:49 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 2 23 theme Proposed 294:301 arg1 functions 303:311 Proposed functions 294:311 Proposed functions of this protein 294:327 Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. 12775711 1 24 theme family 183:188 arg1 glycoprotein 140:151 The 39-kDa human cartilage glycoprotein 113:151 The 39-kDa human cartilage glycoprotein (HCGP39) 113:160 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 24 theme family 183:188 arg1 member 165:170 a member 163:170 a member of a novel family of chitinase-like lectins (Chilectins) 163:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 0 25 theme ligand-induced 14:27 arg1 change 44:49 ligand-induced conformational change 14:49 ligand-induced conformational change 14:49 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 1 26 theme certain 277:283 arg1 cancers 285:291 certain cancers 277:291 certain cancers 277:291 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 5 27 theme physiological 693:705 arg1 ligands 723:729 any physiological or pathological ligands 689:729 any physiological or pathological ligands of the Chi-lectins 689:748 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 9 28 theme chitin-like 1194:1204 arg1 ligands 1222:1228 chitin-like oligosaccharide ligands 1194:1228 chitin-like oligosaccharide ligands 1194:1228 Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. 12775711 1 29 theme 39-kDa 117:122 arg1 glycoprotein 140:151 The 39-kDa human cartilage glycoprotein 113:151 The 39-kDa human cartilage glycoprotein (HCGP39) 113:160 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 29 theme 39-kDa 117:122 arg1 member 165:170 a member 163:170 a member of a novel family of chitinase-like lectins (Chilectins) 163:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 29 theme 39-kDa 117:122 arg1 HCGP39 154:159 HCGP39 154:159 HCGP39 154:159 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 4 30 from surface 620:626 arg1 presence 570:577 the presence 566:577 the presence of a ligand for this protein at the cell surface 566:626 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 4 31 theme cell 615:618 arg1 surface 620:626 the cell surface 611:626 the cell surface 611:626 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 7 32 with complex 936:942 arg1 GlcNAc8 949:955 GlcNAc8 949:955 GlcNAc8 949:955 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 9 33 theme oligosaccharide 1206:1220 arg1 ligands 1222:1228 chitin-like oligosaccharide ligands 1194:1228 chitin-like oligosaccharide ligands 1194:1228 Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. 12775711 6 34 with chitooligosaccharides 841:861 arg1 affinity 879:886 micromolar affinity 868:886 micromolar affinity 868:886 Here, we show that HCGP39 is able to bind chitooligosaccharides with micromolar affinity. 12775711 1 35 theme chitinase-like 193:206 arg1 Chilectins 217:226 Chilectins 217:226 Chilectins 217:226 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 35 theme chitinase-like 193:206 arg1 lectins 208:214 chitinase-like lectins 193:214 chitinase-like lectins (Chilectins) 193:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 5 36 theme protein-ligand 771:784 arg1 interaction 786:796 the protein-ligand interaction 767:796 the protein-ligand interaction 767:796 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 1 37 gly glycoprotein 140:151 arg1 glycoprotein 140:151 The 39-kDa human cartilage glycoprotein 113:151 The 39-kDa human cartilage glycoprotein (HCGP39) 113:160 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 37 gly glycoprotein 140:151 arg1 member 165:170 a member 163:170 a member of a novel family of chitinase-like lectins (Chilectins) 163:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 37 gly glycoprotein 140:151 arg1 HCGP39 154:159 HCGP39 154:159 HCGP39 154:159 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 38 theme lectins 208:214 arg1 family 183:188 a novel family 175:188 a novel family of chitinase-like lectins (Chilectins) 175:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 0 39 theme 39-kDa 58:63 arg1 glycoprotein 65:76 the 39-kDa glycoprotein 54:76 the 39-kDa glycoprotein 54:76 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 8 40 theme oligosaccharide 1080:1094 arg1 ligand 1096:1101 the oligosaccharide ligand 1076:1101 the oligosaccharide ligand 1076:1101 However, unlike the chitinases, binding of the oligosaccharide ligand to HCGP39 induces a large conformational change. 12775711 1 41 theme human 124:128 arg1 glycoprotein 140:151 The 39-kDa human cartilage glycoprotein 113:151 The 39-kDa human cartilage glycoprotein (HCGP39) 113:160 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 41 theme human 124:128 arg1 member 165:170 a member 163:170 a member of a novel family of chitinase-like lectins (Chilectins) 163:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 41 theme human 124:128 arg1 HCGP39 154:159 HCGP39 154:159 HCGP39 154:159 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 5 42 theme pathological 710:721 arg1 ligands 723:729 any physiological or pathological ligands 689:729 any physiological or pathological ligands of the Chi-lectins 689:748 There is currently no information regarding the identity of any physiological or pathological ligands of the Chi-lectins or the nature of the protein-ligand interaction. 12775711 4 43 theme signaling 538:546 arg1 pathways 548:555 cytoplasmic signaling pathways 526:555 cytoplasmic signaling pathways 526:555 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 9 44 theme innate 1259:1264 arg1 responses 1266:1274 innate responses 1259:1274 innate responses to chitinous pathogens, such as fungi and nematodes 1259:1326 Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. 12775711 1 45 theme cartilage 130:138 arg1 glycoprotein 140:151 The 39-kDa human cartilage glycoprotein 113:151 The 39-kDa human cartilage glycoprotein (HCGP39) 113:160 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 45 theme cartilage 130:138 arg1 member 165:170 a member 163:170 a member of a novel family of chitinase-like lectins (Chilectins) 163:227 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 1 45 theme cartilage 130:138 arg1 HCGP39 154:159 HCGP39 154:159 HCGP39 154:159 The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. 12775711 4 46 theme cytoplasmic 526:536 arg1 pathways 548:555 cytoplasmic signaling pathways 526:555 cytoplasmic signaling pathways 526:555 The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. 12775711 7 47 theme family 1011:1016 arg1 chitinases 1021:1030 family 18 chitinases 1011:1030 family 18 chitinases 1011:1030 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 2 48 from defense 380:386 arg1 remodeling 365:374 connective tissue remodeling 347:374 connective tissue remodeling 347:374 Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. 12775711 2 49 theme protein 321:327 arg1 functions 303:311 Proposed functions 294:311 Proposed functions of this protein 294:327 Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. 12775711 7 50 theme native 915:920 arg1 protein 922:928 the native protein 911:928 the native protein 911:928 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 7 51 theme identical 990:998 arg1 fashion 1000:1006 identical fashion 990:1006 identical fashion 990:1006 Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. 12775711 3 52 theme tissue 478:483 arg1 cells 485:489 connective tissue cells 467:489 connective tissue cells 467:489 Similar to other Chi-lectins, HCGP39 promotes the growth of connective tissue cells. 12775711 3 53 theme connective 467:476 arg1 cells 485:489 connective tissue cells 467:489 connective tissue cells 467:489 Similar to other Chi-lectins, HCGP39 promotes the growth of connective tissue cells. 12775711 0 54 from chondrocytes 99:110 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 0 54 from chondrocytes 99:110 arg1 change 44:49 ligand-induced conformational change 14:49 ligand-induced conformational change 14:49 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 2 55 theme tissue 358:363 arg1 remodeling 365:374 connective tissue remodeling 347:374 connective tissue remodeling 347:374 Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. 12775711 0 56 theme glycoprotein 65:76 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 0 56 theme glycoprotein 65:76 arg1 change 44:49 ligand-induced conformational change 14:49 ligand-induced conformational change 14:49 Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. 12775711 2 57 theme connective 347:356 arg1 remodeling 365:374 connective tissue remodeling 347:374 connective tissue remodeling 347:374 Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. 17502376 0 0 theme other 110:114 arg1 substrates 116:125 other substrates 110:125 other substrates 110:125 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 2 1 theme p97-based 465:473 arg1 complex 475:481 a p97-based complex 463:481 a p97-based complex 463:481 Although it is clear that IP(3) receptors are polyubiquitinated upon activation and are transferred to the proteasome by a p97-based complex, currently nothing is known about the proteins that initially select activated IP(3) receptors for ERAD. 17502376 6 2 theme receptor 1212:1219 arg1 polyubiquitination 1221:1238 IP(3) receptor polyubiquitination 1206:1238 IP(3) receptor polyubiquitination 1206:1238 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 7 3 theme substrate 1404:1412 arg1 it 1388:1389 it 1388:1389 it 1388:1389 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 7 3 theme substrate 1404:1412 arg1 factor 1426:1431 a substrate recognition factor 1402:1431 a substrate recognition factor 1402:1431 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 5 4 theme ERAD 1049:1052 arg1 components 1062:1071 ERAD pathway components 1049:1071 ERAD pathway components 1049:1071 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 2 5 theme 3 565:565 arg1 receptors 568:576 activated IP(3) receptors 552:576 activated IP(3) receptors 552:576 Although it is clear that IP(3) receptors are polyubiquitinated upon activation and are transferred to the proteasome by a p97-based complex, currently nothing is known about the proteins that initially select activated IP(3) receptors for ERAD. 17502376 6 6 theme expression 1156:1165 arg1 Suppression 1135:1145 Suppression 1135:1145 Suppression of SPFH2 expression by RNA interference 1135:1185 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 0 7 theme substrates 116:125 arg1 degradation 52:62 the endoplasmic reticulum-associated degradation 15:62 the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells 15:144 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 5 8 theme resting 1006:1012 arg1 cells 1029:1033 resting and stimulated cells 1006:1033 resting and stimulated cells 1006:1033 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 6 9 theme substrates 1289:1298 arg1 degradation 1244:1254 degradation 1244:1254 degradation 1244:1254 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 6 9 theme substrates 1289:1298 arg1 processing 1264:1273 the processing 1260:1273 the processing of other ERAD substrates 1260:1298 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 6 9 theme substrates 1289:1298 arg1 polyubiquitination 1221:1238 IP(3) receptor polyubiquitination 1206:1238 IP(3) receptor polyubiquitination 1206:1238 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 3 10 theme endogenous 675:684 arg1 receptors 702:710 endogenous activated IP(3) receptors 675:710 endogenous activated IP(3) receptors 675:710 Here, we sought to identify novel proteins that associate with and mediate the ERAD of endogenous activated IP(3) receptors. 17502376 7 11 theme ERAD 1348:1351 arg1 SPFH2 1333:1337 SPFH2 1333:1337 SPFH2 1333:1337 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 7 11 theme ERAD 1348:1351 arg1 component 1361:1369 a key ERAD pathway component 1342:1369 a key ERAD pathway component 1342:1369 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 0 12 theme mammalian 130:138 arg1 cells 140:144 mammalian cells 130:144 mammalian cells 130:144 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 0 13 from degradation 52:62 arg1 cells 140:144 mammalian cells 130:144 mammalian cells 130:144 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 6 14 theme ERAD 1284:1287 arg1 substrates 1289:1298 other ERAD substrates 1278:1298 other ERAD substrates 1278:1298 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 7 15 theme pathway 1353:1359 arg1 SPFH2 1333:1337 SPFH2 1333:1337 SPFH2 1333:1337 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 7 15 theme pathway 1353:1359 arg1 component 1361:1369 a key ERAD pathway component 1342:1369 a key ERAD pathway component 1342:1369 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 5 16 dep protein 954:960 arg1 linked 1039:1044 linked 1039:1044 linked 1039:1044 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 4 17 theme proteins 907:914 arg1 polyubiquitination 849:866 significant polyubiquitination 837:866 significant polyubiquitination 837:866 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 4 17 theme proteins 907:914 arg1 association 876:886 the association 872:886 the association of p97 and related proteins 872:914 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 7 18 theme key 1344:1346 arg1 SPFH2 1333:1337 SPFH2 1333:1337 SPFH2 1333:1337 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 7 18 theme key 1344:1346 arg1 component 1361:1369 a key ERAD pathway component 1342:1369 a key ERAD pathway component 1342:1369 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 2 19 theme IP 368:369 arg1 receptors 374:382 IP(3) receptors 368:382 IP(3) receptors 368:382 Although it is clear that IP(3) receptors are polyubiquitinated upon activation and are transferred to the proteasome by a p97-based complex, currently nothing is known about the proteins that initially select activated IP(3) receptors for ERAD. 17502376 0 20 theme endoplasmic 19:29 arg1 degradation 52:62 the endoplasmic reticulum-associated degradation 15:62 the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells 15:144 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 5 21 theme endogenous 1089:1098 arg1 substrates 1100:1109 endogenous substrates 1089:1109 endogenous substrates undergoing degradation 1089:1132 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 6 22 theme other 1278:1282 arg1 substrates 1289:1298 other ERAD substrates 1278:1298 other ERAD substrates 1278:1298 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 1 23 theme reticulum 210:218 arg1 channels 242:249 endoplasmic reticulum (ER) membrane calcium channels 198:249 endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway 198:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 23 theme reticulum 210:218 arg1 receptors 184:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 3 24 theme activated 686:694 arg1 receptors 702:710 endogenous activated IP(3) receptors 675:710 endogenous activated IP(3) receptors 675:710 Here, we sought to identify novel proteins that associate with and mediate the ERAD of endogenous activated IP(3) receptors. 17502376 4 25 theme significant 837:847 arg1 polyubiquitination 849:866 significant polyubiquitination 837:866 significant polyubiquitination 837:866 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 5 26 theme ER 990:991 arg1 lumen 993:997 the ER lumen 986:997 the ER lumen 986:997 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 3 27 theme IP 696:697 arg1 receptors 702:710 endogenous activated IP(3) receptors 675:710 endogenous activated IP(3) receptors 675:710 Here, we sought to identify novel proteins that associate with and mediate the ERAD of endogenous activated IP(3) receptors. 17502376 6 28 theme RNA 1170:1172 arg1 interference 1174:1185 RNA interference 1170:1185 RNA interference 1170:1185 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 1 29 theme endoplasmic 198:208 arg1 ER 221:222 ER 221:222 ER 221:222 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 29 theme endoplasmic 198:208 arg1 reticulum 210:218 endoplasmic reticulum 198:218 endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway 198:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 0 30 theme reticulum-associated 31:50 arg1 degradation 52:62 the endoplasmic reticulum-associated degradation 15:62 the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells 15:144 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 1 31 theme ER-associated 300:312 arg1 degradation 314:324 ER-associated degradation 300:324 the ER-associated degradation (ERAD) pathway 296:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 31 theme ER-associated 300:312 arg1 ERAD 327:330 ERAD 327:330 ERAD 327:330 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 4 32 theme domain-containing 744:760 arg1 protein 762:768 an uncharacterized SPFH domain-containing protein 720:768 an uncharacterized SPFH domain-containing protein 720:768 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 4 32 theme domain-containing 744:760 arg1 SPFH2 713:717 SPFH2 713:717 SPFH2 713:717 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 4 33 theme p97 891:893 arg1 polyubiquitination 849:866 significant polyubiquitination 837:866 significant polyubiquitination 837:866 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 4 33 theme p97 891:893 arg1 association 876:886 the association 872:886 the association of p97 and related proteins 872:914 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 0 34 theme inositol 67:74 arg1 receptors 96:104 inositol 1,4,5-trisphosphate receptors 67:104 inositol 1,4,5-trisphosphate receptors 67:104 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 1 35 theme degradation 314:324 arg1 pathway 333:339 the ER-associated degradation (ERAD) pathway 296:339 the ER-associated degradation (ERAD) pathway 296:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 4 36 theme related 899:905 arg1 proteins 907:914 related proteins 899:914 related proteins 899:914 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 5 37 theme membrane 945:952 arg1 protein 954:960 an ER membrane protein 939:960 an ER membrane protein largely residing within the ER lumen 939:997 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 5 38 theme stimulated 1018:1027 arg1 cells 1029:1033 resting and stimulated cells 1006:1033 resting and stimulated cells 1006:1033 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 5 39 attach linked 1039:1044 arg3 substrates 1100:1109 endogenous substrates 1089:1109 endogenous substrates undergoing degradation 1089:1132 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 2 40 theme activated 552:560 arg1 receptors 568:576 activated IP(3) receptors 552:576 activated IP(3) receptors 552:576 Although it is clear that IP(3) receptors are polyubiquitinated upon activation and are transferred to the proteasome by a p97-based complex, currently nothing is known about the proteins that initially select activated IP(3) receptors for ERAD. 17502376 6 41 theme SPFH2 1150:1154 arg1 expression 1156:1165 SPFH2 expression 1150:1165 SPFH2 expression 1150:1165 Suppression of SPFH2 expression by RNA interference markedly inhibited IP(3) receptor polyubiquitination and degradation and the processing of other ERAD substrates. 17502376 5 42 theme pathway 1054:1060 arg1 components 1062:1071 ERAD pathway components 1049:1071 ERAD pathway components 1049:1071 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 1 43 theme Inositol 147:154 arg1 1,4,5-trisphosphate 156:174 Inositol 1,4,5-trisphosphate 147:174 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 43 theme Inositol 147:154 arg1 IP 177:178 IP(3) 177:181 IP(3) 177:181 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 3 44 theme receptors 702:710 arg1 ERAD 667:670 the ERAD 663:670 the ERAD of endogenous activated IP(3) receptors 663:710 Here, we sought to identify novel proteins that associate with and mediate the ERAD of endogenous activated IP(3) receptors. 17502376 5 45 theme ER 942:943 arg1 protein 954:960 an ER membrane protein 939:960 an ER membrane protein largely residing within the ER lumen 939:997 SPFH2 was found to be an ER membrane protein largely residing within the ER lumen and in resting and stimulated cells was linked to ERAD pathway components, apparently via endogenous substrates undergoing degradation. 17502376 1 46 theme 1,4,5-trisphosphate 156:174 arg1 channels 242:249 endoplasmic reticulum (ER) membrane calcium channels 198:249 endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway 198:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 46 theme 1,4,5-trisphosphate 156:174 arg1 receptors 184:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 47 theme membrane 225:232 arg1 channels 242:249 endoplasmic reticulum (ER) membrane calcium channels 198:249 endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway 198:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 47 theme membrane 225:232 arg1 receptors 184:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 7 48 theme recognition 1414:1424 arg1 it 1388:1389 it 1388:1389 it 1388:1389 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 7 48 theme recognition 1414:1424 arg1 factor 1426:1431 a substrate recognition factor 1402:1431 a substrate recognition factor 1402:1431 Overall, these studies identify SPFH2 as a key ERAD pathway component and suggest that it may act as a substrate recognition factor. 17502376 1 49 theme calcium 234:240 arg1 channels 242:249 endoplasmic reticulum (ER) membrane calcium channels 198:249 endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway 198:339 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 1 49 theme calcium 234:240 arg1 receptors 184:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors 147:192 Inositol 1,4,5-trisphosphate (IP(3)) receptors are endoplasmic reticulum (ER) membrane calcium channels that, upon activation, become substrates for the ER-associated degradation (ERAD) pathway. 17502376 0 50 theme receptors 96:104 arg1 degradation 52:62 the endoplasmic reticulum-associated degradation 15:62 the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells 15:144 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 3 51 theme novel 616:620 arg1 proteins 622:629 novel proteins 616:629 novel proteins that associate with and mediate the ERAD of endogenous activated IP(3) receptors 616:710 Here, we sought to identify novel proteins that associate with and mediate the ERAD of endogenous activated IP(3) receptors. 17502376 4 52 theme SPFH 739:742 arg1 protein 762:768 an uncharacterized SPFH domain-containing protein 720:768 an uncharacterized SPFH domain-containing protein 720:768 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 4 52 theme SPFH 739:742 arg1 SPFH2 713:717 SPFH2 713:717 SPFH2 713:717 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 0 53 theme 1,4,5-trisphosphate 76:94 arg1 receptors 96:104 inositol 1,4,5-trisphosphate receptors 67:104 inositol 1,4,5-trisphosphate receptors 67:104 SPFH2 mediates the endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors and other substrates in mammalian cells. 17502376 4 54 theme uncharacterized 723:737 arg1 protein 762:768 an uncharacterized SPFH domain-containing protein 720:768 an uncharacterized SPFH domain-containing protein 720:768 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 17502376 4 54 theme uncharacterized 723:737 arg1 SPFH2 713:717 SPFH2 713:717 SPFH2 713:717 SPFH2, an uncharacterized SPFH domain-containing protein, rapidly associated with IP(3) receptors in a manner that preceded significant polyubiquitination and the association of p97 and related proteins. 22285761 7 0 theme binding 1150:1156 arg1 properties 1158:1167 the uPA binding properties 1142:1167 the uPA binding properties of uPAR 1142:1175 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 5 1 from structure 698:706 arg1 state 741:745 its ligand-free state 725:745 its ligand-free state 725:745 The crystal structure of human uPAR in its ligand-free state would clarify this issue, but such information remains unfortunately elusive. 22285761 2 2 theme multifunctional 370:384 arg1 role 386:389 this multifunctional role 365:389 this multifunctional role 365:389 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 1 3 theme -mediated 238:246 arg1 fibrinolysis 248:259 urokinase-type plasminogen activator (uPA)-mediated fibrinolysis 196:259 urokinase-type plasminogen activator (uPA)-mediated fibrinolysis 196:259 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 6 4 theme amino-terminal 956:969 arg1 fragment 971:978 amino-terminal fragment 956:978 amino-terminal fragment (ATF) to 3.2 Å 956:993 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 6 4 theme amino-terminal 956:969 arg1 ATF 981:983 ATF 981:983 ATF 981:983 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 0 5 from structure 8:16 arg1 form 61:64 a ligand-free form 47:64 a ligand-free form 47:64 Crystal structure of the urokinase receptor in a ligand-free form. 22285761 7 6 theme wild-type 1064:1072 arg1 complex 1083:1089 the wild-type uPAR·ATF complex 1060:1089 the wild-type uPAR·ATF complex 1060:1089 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 1 7 dep receptor 81:88 arg1 uPAR 137:140 uPAR 137:140 uPAR 137:140 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 7 dep receptor 81:88 arg1 receptor 127:134 urokinase-type plasminogen activator receptor 90:134 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 4 8 theme inherent 593:600 arg1 flexibility 575:585 this flexibility 570:585 this flexibility 570:585 It is, however, not clear whether this flexibility is an inherent property of the uPAR structure per se or whether it is induced upon ligand binding. 22285761 4 8 theme inherent 593:600 arg1 property 602:609 an inherent property 590:609 an inherent property of the uPAR structure 590:631 It is, however, not clear whether this flexibility is an inherent property of the uPAR structure per se or whether it is induced upon ligand binding. 22285761 10 9 theme important 1513:1521 arg1 features 1534:1541 Two important structural features 1509:1541 Two important structural features 1509:1541 Two important structural features are highlighted by the present uPAR structure. 22285761 12 10 theme loop 1661:1664 arg1 130-140 1666:1672 loop 130-140 1661:1672 loop 130-140 of uPAR 1661:1680 Second, loop 130-140 of uPAR plays a dynamic role during ligand loading/unloading. 22285761 9 11 theme DII-DIII 1483:1490 arg1 boundary 1499:1506 the DII-DIII domain boundary 1479:1506 the DII-DIII domain boundary 1479:1506 The domain boundary between uPAR DI-DII domains is more flexible than the DII-DIII domain boundary. 22285761 1 12 theme surface 148:154 arg1 receptor 81:88 The urokinase receptor 67:88 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 12 theme surface 148:154 arg1 receptor 156:163 a surface receptor 146:163 a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis 146:346 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 5 13 theme ligand-free 729:739 arg1 state 741:745 its ligand-free state 725:745 its ligand-free state 725:745 The crystal structure of human uPAR in its ligand-free state would clarify this issue, but such information remains unfortunately elusive. 22285761 11 14 theme domain 1609:1614 arg1 boundary 1616:1623 the DI-DIII domain boundary 1597:1623 the DI-DIII domain boundary 1597:1623 First, the DI-DIII domain boundary may face the cell membrane. 22285761 13 15 theme receptor 1897:1904 arg1 dynamics 1872:1879 the inter-domain dynamics 1855:1879 the inter-domain dynamics of this modular receptor 1855:1904 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 8 16 theme uPAR 1264:1267 arg1 definition 1250:1259 the first structural definition 1229:1259 the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies 1229:1406 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 13 17 theme uPAR 1786:1789 arg1 relationships 1810:1822 uPAR structure-function relationships 1786:1822 uPAR structure-function relationships 1786:1822 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 9 18 theme DI-DII 1442:1447 arg1 domains 1449:1455 uPAR DI-DII domains 1437:1455 uPAR DI-DII domains 1437:1455 The domain boundary between uPAR DI-DII domains is more flexible than the DII-DIII domain boundary. 22285761 3 19 contain possesses 502:510 arg2 flexibility 523:533 structural flexibility 512:533 structural flexibility 512:533 Structural studies suggest that uPAR possesses structural flexibility. 22285761 3 19 contain possesses 502:510 arg1 uPAR 497:500 uPAR 497:500 uPAR 497:500 Structural studies suggest that uPAR possesses structural flexibility. 22285761 12 20 theme dynamic 1690:1696 arg1 role 1698:1701 a dynamic role 1688:1701 a dynamic role 1688:1701 Second, loop 130-140 of uPAR plays a dynamic role during ligand loading/unloading. 22285761 6 21 with complex 943:949 arg1 fragment 971:978 amino-terminal fragment 956:978 amino-terminal fragment (ATF) to 3.2 Å 956:993 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 6 21 with complex 943:949 arg1 ATF 981:983 ATF 981:983 ATF 981:983 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 13 22 theme dynamics 1872:1879 arg1 importance 1841:1850 the importance 1837:1850 the importance of the inter-domain dynamics of this modular receptor 1837:1904 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 2 23 theme extracellular 411:423 arg1 vitronectin 452:462 vitronectin 452:462 vitronectin 452:462 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 2 23 theme extracellular 411:423 arg1 ligands 425:431 several extracellular ligands 403:431 several extracellular ligands 403:431 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 2 23 theme extracellular 411:423 arg1 uPA 444:446 uPA 444:446 uPA 444:446 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 6 24 theme human 879:883 arg1 uPAR 885:888 a stabilized, human uPAR 865:888 uPAR 885:888 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 6 24 theme human 879:883 arg1 H47C/N259C 891:900 H47C/N259C 891:900 H47C/N259C 891:900 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 10 25 theme uPAR 1574:1577 arg1 structure 1579:1587 the present uPAR structure 1562:1587 the present uPAR structure 1562:1587 Two important structural features are highlighted by the present uPAR structure. 22285761 4 26 theme uPAR 618:621 arg1 structure 623:631 the uPAR structure 614:631 the uPAR structure 614:631 It is, however, not clear whether this flexibility is an inherent property of the uPAR structure per se or whether it is induced upon ligand binding. 22285761 1 27 theme capable 165:171 arg1 receptor 81:88 The urokinase receptor 67:88 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 27 theme capable 165:171 arg1 receptor 156:163 a surface receptor 146:163 a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis 146:346 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 11 28 theme DI-DIII 1601:1607 arg1 boundary 1616:1623 the DI-DIII domain boundary 1597:1623 the DI-DIII domain boundary 1597:1623 First, the DI-DIII domain boundary may face the cell membrane. 22285761 9 29 theme domain 1492:1497 arg1 boundary 1499:1506 the DII-DIII domain boundary 1479:1506 the DII-DIII domain boundary 1479:1506 The domain boundary between uPAR DI-DII domains is more flexible than the DII-DIII domain boundary. 22285761 1 30 theme urokinase 71:79 arg1 receptor 81:88 The urokinase receptor 67:88 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 30 theme urokinase 71:79 arg1 receptor 156:163 a surface receptor 146:163 a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis 146:346 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 6 31 theme uPAR 885:888 arg1 structures 851:860 the crystal structures 839:860 the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å 839:993 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 0 32 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the urokinase receptor in a ligand-free form. 0:65 Crystal structure of the urokinase receptor in a ligand-free form. 22285761 1 33 theme pericellular 268:279 arg1 micro-environment 281:297 the pericellular micro-environment 264:297 the pericellular micro-environment 264:297 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 8 34 from definition 1250:1259 arg1 form 1288:1291 its ligand-free form 1272:1291 its ligand-free form 1272:1291 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 6 35 from structures 851:860 arg1 form 922:925 its ligand-free form 906:925 its ligand-free form to 2.4 Å 906:934 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 6 35 from structures 851:860 arg1 complex 943:949 complex 943:949 complex with amino-terminal fragment (ATF) to 3.2 Å 943:993 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 4 36 theme ligand 670:675 arg1 binding 677:683 ligand binding 670:683 ligand binding 670:683 It is, however, not clear whether this flexibility is an inherent property of the uPAR structure per se or whether it is induced upon ligand binding. 22285761 8 37 theme first 1233:1237 arg1 definition 1250:1259 the first structural definition 1229:1259 the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies 1229:1406 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 8 38 theme extensive 1378:1386 arg1 studies 1400:1406 extensive biochemical studies 1378:1406 extensive biochemical studies 1378:1406 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 1 39 theme urokinase-type 90:103 arg1 uPAR 137:140 uPAR 137:140 uPAR 137:140 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 39 theme urokinase-type 90:103 arg1 receptor 127:134 urokinase-type plasminogen activator receptor 90:134 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 13 40 theme new 1768:1770 arg1 insights 1772:1779 new insights 1768:1779 new insights into uPAR structure-function relationships 1768:1822 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 8 41 theme uPAR 1203:1206 arg1 structure 1190:1198 The present structure 1178:1198 The present structure of uPAR(H47C/N259C) 1178:1218 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 10 42 theme structural 1523:1532 arg1 features 1534:1541 Two important structural features 1509:1541 Two important structural features 1509:1541 Two important structural features are highlighted by the present uPAR structure. 22285761 13 43 theme modular 1889:1895 arg1 receptor 1897:1904 this modular receptor 1884:1904 this modular receptor 1884:1904 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 5 44 theme human 711:715 arg1 uPAR 717:720 human uPAR 711:720 human uPAR 711:720 The crystal structure of human uPAR in its ligand-free state would clarify this issue, but such information remains unfortunately elusive. 22285761 6 45 theme stabilized 867:876 arg1 uPAR 885:888 a stabilized, human uPAR 865:888 uPAR 885:888 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 6 45 theme stabilized 867:876 arg1 H47C/N259C 891:900 H47C/N259C 891:900 H47C/N259C 891:900 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 1 46 theme plasminogen 105:115 arg1 uPAR 137:140 uPAR 137:140 uPAR 137:140 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 46 theme plasminogen 105:115 arg1 receptor 127:134 urokinase-type plasminogen activator receptor 90:134 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 0 47 theme receptor 35:42 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the urokinase receptor in a ligand-free form. 0:65 Crystal structure of the urokinase receptor in a ligand-free form. 22285761 13 48 theme structure-function 1791:1808 arg1 relationships 1810:1822 uPAR structure-function relationships 1786:1822 uPAR structure-function relationships 1786:1822 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 5 49 theme uPAR 717:720 arg1 structure 698:706 The crystal structure 686:706 The crystal structure of human uPAR in its ligand-free state 686:745 The crystal structure of human uPAR in its ligand-free state would clarify this issue, but such information remains unfortunately elusive. 22285761 11 50 theme cell 1638:1641 arg1 membrane 1643:1650 the cell membrane 1634:1650 the cell membrane 1634:1650 First, the DI-DIII domain boundary may face the cell membrane. 22285761 8 51 theme present 1182:1188 arg1 structure 1190:1198 The present structure 1178:1198 The present structure of uPAR(H47C/N259C) 1178:1218 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 1 52 theme urokinase-type 196:209 arg1 uPA 234:236 uPA 234:236 uPA 234:236 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 52 theme urokinase-type 196:209 arg1 activator 223:231 urokinase-type plasminogen activator 196:231 urokinase-type plasminogen activator (uPA) 196:237 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 0 53 theme urokinase 25:33 arg1 receptor 35:42 the urokinase receptor 21:42 the urokinase receptor 21:42 Crystal structure of the urokinase receptor in a ligand-free form. 22285761 5 54 theme such 777:780 arg1 information 782:792 such information 777:792 such information 777:792 The crystal structure of human uPAR in its ligand-free state would clarify this issue, but such information remains unfortunately elusive. 22285761 8 55 theme biochemical 1388:1398 arg1 studies 1400:1406 extensive biochemical studies 1378:1406 extensive biochemical studies 1378:1406 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 1 56 theme plasminogen 211:221 arg1 uPA 234:236 uPA 234:236 uPA 234:236 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 56 theme plasminogen 211:221 arg1 activator 223:231 urokinase-type plasminogen activator 196:231 urokinase-type plasminogen activator (uPA) 196:237 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 7 57 theme uPAR·ATF 1074:1081 arg1 complex 1083:1089 the wild-type uPAR·ATF complex 1060:1089 the wild-type uPAR·ATF complex 1060:1089 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 1 58 theme cell 318:321 arg1 migration 323:331 cell migration 318:331 cell migration 318:331 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 2 59 with Consistent 349:358 arg1 role 386:389 this multifunctional role 365:389 this multifunctional role 365:389 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 8 60 theme active 1335:1340 arg1 conformations 1342:1354 the biologically active conformations 1318:1354 the biologically active conformations of uPAR 1318:1362 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 1 61 theme activator 117:125 arg1 uPAR 137:140 uPAR 137:140 uPAR 137:140 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 1 61 theme activator 117:125 arg1 receptor 127:134 urokinase-type plasminogen activator receptor 90:134 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) 67:141 The urokinase receptor urokinase-type plasminogen activator receptor (uPAR) is a surface receptor capable of not only focalizing urokinase-type plasminogen activator (uPA)-mediated fibrinolysis to the pericellular micro-environment but also promoting cell migration and chemotaxis. 22285761 7 62 theme uPAR 1013:1016 arg1 structure 1000:1008 The structure 996:1008 The structure of uPAR(H47C/N259C) in complex with ATF 996:1048 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 9 63 theme uPAR 1437:1440 arg1 domains 1449:1455 uPAR DI-DII domains 1437:1455 uPAR DI-DII domains 1437:1455 The domain boundary between uPAR DI-DII domains is more flexible than the DII-DIII domain boundary. 22285761 6 64 theme crystal 843:849 arg1 structures 851:860 the crystal structures 839:860 the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å 839:993 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 8 65 theme uPAR 1359:1362 arg1 conformations 1342:1354 the biologically active conformations 1318:1354 the biologically active conformations of uPAR 1318:1362 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 6 66 theme ligand-free 910:920 arg1 form 922:925 its ligand-free form 906:925 its ligand-free form to 2.4 Å 906:934 We now report the crystal structures of a stabilized, human uPAR (H47C/N259C) in its ligand-free form to 2.4 Å and in complex with amino-terminal fragment (ATF) to 3.2 Å. 22285761 5 67 theme crystal 690:696 arg1 structure 698:706 The crystal structure 686:706 The crystal structure of human uPAR in its ligand-free state 686:745 The crystal structure of human uPAR in its ligand-free state would clarify this issue, but such information remains unfortunately elusive. 22285761 7 68 with complex 1033:1039 arg1 ATF 1046:1048 ATF 1046:1048 ATF 1046:1048 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 13 69 theme inter-domain 1859:1870 arg1 dynamics 1872:1879 the inter-domain dynamics 1855:1879 the inter-domain dynamics of this modular receptor 1855:1904 Together, these studies provide new insights into uPAR structure-function relationships, emphasizing the importance of the inter-domain dynamics of this modular receptor. 22285761 4 70 theme structure 623:631 arg1 flexibility 575:585 this flexibility 570:585 this flexibility 570:585 It is, however, not clear whether this flexibility is an inherent property of the uPAR structure per se or whether it is induced upon ligand binding. 22285761 4 70 theme structure 623:631 arg1 property 602:609 an inherent property 590:609 an inherent property of the uPAR structure 590:631 It is, however, not clear whether this flexibility is an inherent property of the uPAR structure per se or whether it is induced upon ligand binding. 22285761 8 71 theme ligand-free 1276:1286 arg1 form 1288:1291 its ligand-free form 1272:1291 its ligand-free form 1272:1291 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 8 72 theme conformations 1342:1354 arg1 conformations 1342:1354 the biologically active conformations 1318:1354 the biologically active conformations of uPAR 1318:1362 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 8 72 theme conformations 1342:1354 arg1 one 1311:1313 one 1311:1313 one 1311:1313 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 12 73 theme ligand 1710:1715 arg1 loading/unloading 1717:1733 ligand loading/unloading 1710:1733 ligand loading/unloading 1710:1733 Second, loop 130-140 of uPAR plays a dynamic role during ligand loading/unloading. 22285761 3 74 theme structural 512:521 arg1 flexibility 523:533 structural flexibility 512:533 structural flexibility 512:533 Structural studies suggest that uPAR possesses structural flexibility. 22285761 12 75 theme uPAR 1677:1680 arg1 130-140 1666:1672 loop 130-140 1661:1672 loop 130-140 of uPAR 1661:1680 Second, loop 130-140 of uPAR plays a dynamic role during ligand loading/unloading. 22285761 3 76 theme Structural 465:474 arg1 studies 476:482 Structural studies 465:482 Structural studies 465:482 Structural studies suggest that uPAR possesses structural flexibility. 22285761 7 77 theme uPAR 1172:1175 arg1 properties 1158:1167 the uPA binding properties 1142:1167 the uPA binding properties of uPAR 1142:1175 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 9 78 theme domain 1413:1418 arg1 boundary 1420:1427 The domain boundary 1409:1427 The domain boundary between uPAR DI-DII domains 1409:1455 The domain boundary between uPAR DI-DII domains is more flexible than the DII-DIII domain boundary. 22285761 9 78 theme domain 1413:1418 arg1 flexible 1465:1472 flexible 1465:1472 flexible 1465:1472 The domain boundary between uPAR DI-DII domains is more flexible than the DII-DIII domain boundary. 22285761 2 79 theme several 403:409 arg1 vitronectin 452:462 vitronectin 452:462 vitronectin 452:462 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 2 79 theme several 403:409 arg1 ligands 425:431 several extracellular ligands 403:431 several extracellular ligands 403:431 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 2 79 theme several 403:409 arg1 uPA 444:446 uPA 444:446 uPA 444:446 Consistent with this multifunctional role, uPAR binds several extracellular ligands, including uPA and vitronectin. 22285761 0 80 theme ligand-free 49:59 arg1 form 61:64 a ligand-free form 47:64 a ligand-free form 47:64 Crystal structure of the urokinase receptor in a ligand-free form. 22285761 7 81 from structure 1000:1008 arg1 complex 1033:1039 complex 1033:1039 complex with ATF 1033:1048 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 22285761 8 82 theme structural 1239:1248 arg1 definition 1250:1259 the first structural definition 1229:1259 the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies 1229:1406 The present structure of uPAR(H47C/N259C) provides the first structural definition of uPAR in its ligand-free form, which represents one of the biologically active conformations of uPAR as defined by extensive biochemical studies. 22285761 10 83 theme present 1566:1572 arg1 structure 1579:1587 the present uPAR structure 1562:1587 the present uPAR structure 1562:1587 Two important structural features are highlighted by the present uPAR structure. 22285761 7 84 theme uPA 1146:1148 arg1 properties 1158:1167 the uPA binding properties 1142:1167 the uPA binding properties of uPAR 1142:1175 The structure of uPAR(H47C/N259C) in complex with ATF resembles the wild-type uPAR·ATF complex, demonstrating that these mutations do not perturb the uPA binding properties of uPAR. 18697946 7 0 from elsewhere 1326:1334 arg1 molecules 1343:1351 the molecules 1339:1351 the molecules 1339:1351 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 2 1 theme several 283:289 arg1 isotypes 297:304 several other isotypes 283:304 several other isotypes that use the same alpha chain but have beta chains encoded by other genes 283:378 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 6 2 contain has 1006:1008 arg2 features 1022:1029 some unique features 1010:1029 some unique features 1010:1029 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 6 2 contain has 1006:1008 arg1 molecule 920:927 The molecule 916:927 The molecule 916:927 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 7 3 theme major 1086:1090 arg1 b 1117:1117 b 1117:1117 b 1117:1117 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 3 theme major 1086:1090 arg1 c 1124:1124 c 1124:1124 c 1124:1124 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 3 theme major 1086:1090 arg1 DR52a 1110:1114 DR52a 1110:1114 DR52a 1110:1114 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 3 theme major 1086:1090 arg1 alleles 1101:1107 the three major HLA-DBR3 alleles 1076:1107 the three major HLA-DBR3 alleles (DR52a, b, and c) 1076:1125 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 4 theme no 1309:1310 arg1 polymorphisms 1312:1324 left virtually no polymorphisms 1294:1324 left virtually no polymorphisms 1294:1324 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 6 5 theme other 973:977 arg1 molecules 992:1000 other MHC class II molecules 973:1000 other MHC class II molecules 973:1000 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 1 6 theme CD4 146:148 arg1 cells 155:159 CD4(+) T cells 146:159 CD4(+) T cells 146:159 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 4 7 theme specific 673:680 arg1 diseases 693:700 specific autoimmune diseases 673:700 specific autoimmune diseases 673:700 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 7 8 dep one 1164:1166 arg1 another 1168:1174 another 1168:1174 another 1168:1174 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 5 9 theme HLA-DRB3 838:845 arg1 chains 830:835 these alternate beta chains 809:835 these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 809:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 9 theme HLA-DRB3 838:845 arg1 * 846:846 HLA-DRB3*0301 838:850 HLA-DRB3*0301 838:850 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 1 10 theme complex 100:106 arg1 molecules 116:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 5 11 theme alternate 815:823 arg1 chains 830:835 these alternate beta chains 809:835 these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 809:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 11 theme alternate 815:823 arg1 * 846:846 HLA-DRB3*0301 838:850 HLA-DRB3*0301 838:850 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 6 12 theme unique 1015:1020 arg1 features 1022:1029 some unique features 1010:1029 some unique features 1010:1029 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 6 13 theme peptide-binding 1038:1052 arg1 groove 1054:1059 the peptide-binding groove 1034:1059 the peptide-binding groove 1034:1059 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 4 14 theme HLA-DRB1 589:596 arg1 alleles 598:604 the HLA-DRB1 alleles 585:604 the HLA-DRB1 alleles 585:604 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 6 15 theme class 983:987 arg1 molecules 992:1000 other MHC class II molecules 973:1000 other MHC class II molecules 973:1000 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 7 16 theme major 1224:1228 arg1 pockets 1246:1252 the four major peptide-binding pockets 1215:1252 the four major peptide-binding pockets 1215:1252 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 2 17 dep isotypes 297:304 arg1 HLA-DR 224:229 HLA-DR 224:229 HLA-DR 224:229 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 17 dep isotypes 297:304 arg1 isotype 215:221 the most commonly studied human MHCII isotype 177:221 the most commonly studied human MHCII isotype 177:221 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 18 theme same 319:322 arg1 chain 330:334 the same alpha chain 315:334 the same alpha chain 315:334 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 1 19 theme Class 66:70 arg1 molecules 116:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 3 20 theme antigen-presenting 428:445 arg1 cells 447:451 antigen-presenting cells 428:451 antigen-presenting cells 428:451 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 4 21 theme alternate 624:632 arg1 molecules 637:645 these alternate DR molecules 618:645 these alternate DR molecules 618:645 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 7 22 theme elsewhere 1326:1334 arg1 positions 1265:1273 peptide positions 1257:1273 peptide positions 1, 4, 6, and 9 1257:1288 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 23 attach derived 1151:1157 arg1 one 1164:1166 one 1164:1166 one 1164:1166 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 23 attach derived 1151:1157 arg2 they 1141:1144 they 1141:1144 they 1141:1144 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 1 24 theme major 75:79 arg1 molecules 116:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 7 25 dep alleles 1101:1107 arg1 b 1117:1117 b 1117:1117 b 1117:1117 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 25 dep alleles 1101:1107 arg1 c 1124:1124 c 1124:1124 c 1124:1124 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 25 dep alleles 1101:1107 arg1 DR52a 1110:1114 DR52a 1110:1114 DR52a 1110:1114 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 25 dep alleles 1101:1107 arg1 alleles 1101:1107 the three major HLA-DBR3 alleles 1076:1107 the three major HLA-DBR3 alleles (DR52a, b, and c) 1076:1125 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 1 26 theme T 153:153 arg1 cells 155:159 CD4(+) T cells 146:159 CD4(+) T cells 146:159 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 0 27 theme HLA-DR52c 17:25 arg1 structure 4:12 The structure 0:12 The structure of HLA-DR52c 0:25 The structure of HLA-DR52c: comparison to other HLA-DRB3 alleles. 18697946 7 28 theme peptide 1257:1263 arg1 positions 1265:1273 peptide positions 1257:1273 peptide positions 1, 4, 6, and 9 1257:1288 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 1 29 theme histocompatibility 81:98 arg1 molecules 116:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 4 30 theme cell 708:711 arg1 hypersensitivity 713:728 T cell hypersensitivity 706:728 T cell hypersensitivity 706:728 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 2 31 theme beta 345:348 arg1 chains 350:355 beta chains 345:355 beta chains encoded by other genes 345:378 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 5 32 theme beta 825:828 arg1 chains 830:835 these alternate beta chains 809:835 these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 809:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 32 theme beta 825:828 arg1 * 846:846 HLA-DRB3*0301 838:850 HLA-DRB3*0301 838:850 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 33 theme chains 830:835 arg1 chains 830:835 these alternate beta chains 809:835 these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 809:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 33 theme chains 830:835 arg1 one 802:804 one 802:804 one 802:804 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 33 theme chains 830:835 arg1 * 846:846 HLA-DRB3*0301 838:850 HLA-DRB3*0301 838:850 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 2 34 theme beta 238:241 arg1 chain 243:247 chain 243:247 chain 243:247 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 6 35 theme MHC 979:981 arg1 molecules 992:1000 other MHC class II molecules 973:1000 other MHC class II molecules 973:1000 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 2 36 theme HLA-DRB1 267:274 arg1 locus 276:280 the HLA-DRB1 locus 263:280 the HLA-DRB1 locus 263:280 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 3 37 theme other 557:561 arg1 cells 565:569 other T cells 557:569 other T cells 557:569 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 3 38 theme T 509:509 arg1 cells 511:515 T cells 509:515 T cells 509:515 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 5 39 theme HLA-DR 767:772 arg1 DR52c 784:788 DR52c 784:788 DR52c 784:788 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 39 theme HLA-DR 767:772 arg1 molecule 774:781 an HLA-DR molecule 764:781 an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 764:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 3 40 theme peptide 485:491 arg1 presentation 493:504 peptide presentation 485:504 peptide presentation to T cells 485:515 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 6 41 theme conserved 949:957 arg1 elements 959:966 structurally conserved elements 936:966 structurally conserved elements 936:966 The molecule shares structurally conserved elements with other MHC class II molecules but has some unique features in the peptide-binding groove. 18697946 7 42 theme peptide-binding 1230:1244 arg1 pockets 1246:1252 the four major peptide-binding pockets 1215:1252 the four major peptide-binding pockets 1215:1252 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 4 43 theme T 706:706 arg1 hypersensitivity 713:728 T cell hypersensitivity 706:728 T cell hypersensitivity 706:728 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 2 44 theme alpha 324:328 arg1 chain 330:334 the same alpha chain 315:334 the same alpha chain 315:334 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 0 45 theme other 42:46 arg1 alleles 57:63 other HLA-DRB3 alleles 42:63 other HLA-DRB3 alleles 42:63 The structure of HLA-DR52c: comparison to other HLA-DRB3 alleles. 18697946 7 46 theme alleles 1101:1107 arg1 Comparison 1062:1071 Comparison 1062:1071 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) 1062:1125 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 0 47 dep comparison 28:37 arg1 structure 4:12 The structure 0:12 The structure of HLA-DR52c 0:25 The structure of HLA-DR52c: comparison to other HLA-DRB3 alleles. 18697946 2 48 theme other 368:372 arg1 genes 374:378 other genes 368:378 other genes 368:378 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 5 49 theme elongation 897:906 arg1 factor 908:913 the Tu elongation factor 890:913 the Tu elongation factor 890:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 50 theme Tu 894:895 arg1 factor 908:913 the Tu elongation factor 890:913 the Tu elongation factor 890:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 1 51 theme MHCII 109:113 arg1 molecules 116:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules 66:124 Class II major histocompatibility complex (MHCII) molecules present antigens to CD4(+) T cells. 18697946 7 52 theme recombination 1179:1191 arg1 events 1193:1198 recombination events 1179:1198 recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules 1179:1351 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 4 53 theme autoimmune 682:691 arg1 diseases 693:700 specific autoimmune diseases 673:700 specific autoimmune diseases 673:700 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 5 54 contain containing 791:800 arg1 DR52c 784:788 DR52c 784:788 DR52c 784:788 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 54 contain containing 791:800 arg1 molecule 774:781 an HLA-DR molecule 764:781 an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 764:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 54 contain containing 791:800 arg2 one 802:804 one 802:804 one 802:804 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 54 contain containing 791:800 arg2 * 846:846 HLA-DRB3*0301 838:850 HLA-DRB3*0301 838:850 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 54 contain containing 791:800 arg2 chains 830:835 these alternate beta chains 809:835 these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 809:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 3 55 theme other 387:391 arg1 molecules 396:404 These other DR molecules 381:404 These other DR molecules 381:404 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 4 56 theme molecules 637:645 arg1 several 607:613 several 607:613 several 607:613 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 3 57 theme T 563:563 arg1 cells 565:569 other T cells 557:569 other T cells 557:569 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 2 58 theme MHCII 209:213 arg1 HLA-DR 224:229 HLA-DR 224:229 HLA-DR 224:229 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 58 theme MHCII 209:213 arg1 isotype 215:221 the most commonly studied human MHCII isotype 177:221 the most commonly studied human MHCII isotype 177:221 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 4 59 theme DR 634:635 arg1 molecules 637:645 these alternate DR molecules 618:645 these alternate DR molecules 618:645 Like some of the HLA-DRB1 alleles, several of these alternate DR molecules have been associated with specific autoimmune diseases and T cell hypersensitivity. 18697946 3 60 theme DR 393:394 arg1 molecules 396:404 These other DR molecules 381:404 These other DR molecules 381:404 These other DR molecules also are expressed in antigen-presenting cells and are known to participate in peptide presentation to T cells and to be recognized as alloantigens by other T cells. 18697946 5 61 attach derived 877:883 arg2 self-peptide 864:875 a self-peptide 862:875 a self-peptide derived from the Tu elongation factor 862:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 5 61 attach derived 877:883 arg1 factor 908:913 the Tu elongation factor 890:913 the Tu elongation factor 890:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 7 62 theme HLA-DBR3 1092:1099 arg1 b 1117:1117 b 1117:1117 b 1117:1117 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 62 theme HLA-DBR3 1092:1099 arg1 c 1124:1124 c 1124:1124 c 1124:1124 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 62 theme HLA-DBR3 1092:1099 arg1 DR52a 1110:1114 DR52a 1110:1114 DR52a 1110:1114 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 7 62 theme HLA-DBR3 1092:1099 arg1 alleles 1101:1107 the three major HLA-DBR3 alleles 1076:1107 the three major HLA-DBR3 alleles (DR52a, b, and c) 1076:1125 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 2 63 contain have 340:343 arg2 chains 350:355 beta chains 345:355 beta chains encoded by other genes 345:378 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 63 contain have 340:343 arg1 isotypes 297:304 several other isotypes 283:304 several other isotypes that use the same alpha chain but have beta chains encoded by other genes 283:378 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 64 theme human 203:207 arg1 HLA-DR 224:229 HLA-DR 224:229 HLA-DR 224:229 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 64 theme human 203:207 arg1 isotype 215:221 the most commonly studied human MHCII isotype 177:221 the most commonly studied human MHCII isotype 177:221 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 5 65 theme molecule 774:781 arg1 structure 751:759 the structure 747:759 the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor 747:913 Here we present the structure of an HLA-DR molecule (DR52c) containing one of these alternate beta chains (HLA-DRB3*0301) bound to a self-peptide derived from the Tu elongation factor. 18697946 0 66 theme HLA-DRB3 48:55 arg1 alleles 57:63 other HLA-DRB3 alleles 42:63 other HLA-DRB3 alleles 42:63 The structure of HLA-DR52c: comparison to other HLA-DRB3 alleles. 18697946 7 67 theme left 1294:1297 arg1 polymorphisms 1312:1324 left virtually no polymorphisms 1294:1324 left virtually no polymorphisms 1294:1324 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 18697946 2 68 theme studied 195:201 arg1 HLA-DR 224:229 HLA-DR 224:229 HLA-DR 224:229 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 68 theme studied 195:201 arg1 isotype 215:221 the most commonly studied human MHCII isotype 177:221 the most commonly studied human MHCII isotype 177:221 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 2 69 theme other 291:295 arg1 isotypes 297:304 several other isotypes 283:304 several other isotypes that use the same alpha chain but have beta chains encoded by other genes 283:378 In addition to the most commonly studied human MHCII isotype, HLA-DR, whose beta chain is encoded by the HLA-DRB1 locus, several other isotypes that use the same alpha chain but have beta chains encoded by other genes. 18697946 7 70 from molecules 1343:1351 arg1 elsewhere 1326:1334 elsewhere 1326:1334 elsewhere 1326:1334 Comparison of the three major HLA-DBR3 alleles (DR52a, b, and c) suggests that they were derived from one another by recombination events that scrambled the four major peptide-binding pockets at peptide positions 1, 4, 6, and 9 but left virtually no polymorphisms elsewhere in the molecules. 1451807 1 0 gly O-glycosylation 65:79 arg1 Thr7 84:87 Thr7 84:87 Thr7 84:87 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 1 0 gly O-glycosylation 65:79 arg2 Thr7 84:87 Thr7 84:87 Thr7 84:87 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 4 1 from O-glycosylated 495:508 arg1 addition 459:466 addition 459:466 addition 459:466 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 2 2 from heterogeneous 270:282 arg1 mass 297:300 molecular mass 287:300 molecular mass 287:300 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 5 3 theme O-linked 770:777 arg1 carbohydrates 779:791 O-linked carbohydrates 770:791 O-linked carbohydrates 770:791 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 3 4 from Asn62 395:399 arg1 N-glycosylated 377:390 N-glycosylated 377:390 N-glycosylated 377:390 All components are N-glycosylated at Asn62; N-glycosylation does not contribute to heterogeneity. 1451807 6 5 theme amino 828:832 arg1 position 839:846 amino acid position 26 828:849 amino acid position 26 (threonine/asparagine) 828:872 Two allelic variants differing in amino acid position 26 (threonine/asparagine) were identified. 1451807 6 5 theme amino 828:832 arg1 threonine/asparagine 852:871 threonine/asparagine 852:871 threonine/asparagine 852:871 Two allelic variants differing in amino acid position 26 (threonine/asparagine) were identified. 1451807 2 6 from mass 297:300 arg1 heterogeneous 270:282 heterogeneous 270:282 heterogeneous 270:282 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 7 with heterogeneous 270:282 arg1 components 314:323 seven components 308:323 seven components resolved by gel electrophoresis 308:355 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 5 8 theme N-proximal 722:731 arg1 region 733:738 the N-proximal region 718:738 the N-proximal region 718:738 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 4 9 theme neuraminic 584:593 arg1 acid 595:598 neuraminic acid 584:598 neuraminic acid 584:598 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 2 10 theme human 145:149 arg1 TNF-beta 179:186 TNF-beta 179:186 TNF-beta 179:186 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 10 theme human 145:149 arg1 beta 173:176 Natural human tumor necrosis factor beta 137:176 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line 137:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 3 11 gly N-glycosylated 377:390 arg1 components 362:371 All components 358:371 All components 358:371 All components are N-glycosylated at Asn62; N-glycosylation does not contribute to heterogeneity. 1451807 3 11 gly N-glycosylated 377:390 arg2 Asn62 395:399 Asn62 395:399 Asn62 395:399 All components are N-glycosylated at Asn62; N-glycosylation does not contribute to heterogeneity. 1451807 5 12 from cleavage 706:713 arg1 region 733:738 the N-proximal region 718:738 the N-proximal region 718:738 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 2 13 theme Natural 137:143 arg1 TNF-beta 179:186 TNF-beta 179:186 TNF-beta 179:186 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 13 theme Natural 137:143 arg1 beta 173:176 Natural human tumor necrosis factor beta 137:176 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line 137:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 5 14 theme lower 610:614 arg1 forms 631:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 1 15 theme Variable 56:63 arg1 O-glycosylation 65:79 Variable O-glycosylation 56:79 Variable O-glycosylation at Thr7 56:87 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 5 16 theme molecular 616:624 arg1 forms 631:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 6 17 theme allelic 798:804 arg1 variants 806:813 Two allelic variants 794:813 Two allelic variants differing in amino acid position 26 (threonine/asparagine) 794:872 Two allelic variants differing in amino acid position 26 (threonine/asparagine) were identified. 1451807 0 18 theme human 8:12 arg1 beta 36:39 Natural human tumor necrosis factor beta 0:39 Natural human tumor necrosis factor beta (lymphotoxin). 0:54 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 0 18 theme human 8:12 arg1 lymphotoxin 42:52 lymphotoxin 42:52 lymphotoxin 42:52 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 5 19 theme mass 626:629 arg1 forms 631:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 4 20 gly O-glycosylated 495:508 arg1 part 469:472 part 469:472 part of the molecules 469:489 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 4 20 gly O-glycosylated 495:508 arg1 addition 459:466 addition 459:466 addition 459:466 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 4 20 gly O-glycosylated 495:508 arg2 Thr7 513:516 Thr7 513:516 Thr7 513:516 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 4 20 gly O-glycosylated 495:508 arg2 part 469:472 part 469:472 part of the molecules 469:489 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 0 21 theme Natural 0:6 arg1 beta 36:39 Natural human tumor necrosis factor beta 0:39 Natural human tumor necrosis factor beta (lymphotoxin). 0:54 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 0 21 theme Natural 0:6 arg1 lymphotoxin 42:52 lymphotoxin 42:52 lymphotoxin 42:52 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 4 22 from addition 459:466 arg1 O-glycosylated 495:508 O-glycosylated 495:508 O-glycosylated 495:508 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 2 23 theme molecular 287:295 arg1 mass 297:300 molecular mass 287:300 molecular mass 287:300 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 0 24 theme necrosis 20:27 arg1 beta 36:39 Natural human tumor necrosis factor beta 0:39 Natural human tumor necrosis factor beta (lymphotoxin). 0:54 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 0 24 theme necrosis 20:27 arg1 lymphotoxin 42:52 lymphotoxin 42:52 lymphotoxin 42:52 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 4 25 with decoration 568:577 arg1 acid 595:598 neuraminic acid 584:598 neuraminic acid 584:598 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 5 26 theme proteolytic 694:704 arg1 cleavage 706:713 trypsin-like proteolytic cleavage 681:713 trypsin-like proteolytic cleavage in the N-proximal region 681:738 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 0 27 theme tumor 14:18 arg1 beta 36:39 Natural human tumor necrosis factor beta 0:39 Natural human tumor necrosis factor beta (lymphotoxin). 0:54 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 0 27 theme tumor 14:18 arg1 lymphotoxin 42:52 lymphotoxin 42:52 lymphotoxin 42:52 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 1 28 theme proteolytic 90:100 arg1 processing 102:111 proteolytic processing 90:111 proteolytic processing 90:111 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 2 29 theme factor 166:171 arg1 TNF-beta 179:186 TNF-beta 179:186 TNF-beta 179:186 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 29 theme factor 166:171 arg1 beta 173:176 Natural human tumor necrosis factor beta 137:176 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line 137:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 0 30 theme factor 29:34 arg1 beta 36:39 Natural human tumor necrosis factor beta 0:39 Natural human tumor necrosis factor beta (lymphotoxin). 0:54 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 0 30 theme factor 29:34 arg1 lymphotoxin 42:52 lymphotoxin 42:52 lymphotoxin 42:52 Natural human tumor necrosis factor beta (lymphotoxin). 1451807 4 31 theme variable 559:566 arg1 decoration 568:577 variable decoration 559:577 variable decoration with neuraminic acid 559:598 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 2 32 theme necrosis 157:164 arg1 TNF-beta 179:186 TNF-beta 179:186 TNF-beta 179:186 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 32 theme necrosis 157:164 arg1 beta 173:176 Natural human tumor necrosis factor beta 137:176 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line 137:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 4 33 from Thr7 513:516 arg1 O-glycosylated 495:508 O-glycosylated 495:508 O-glycosylated 495:508 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 5 34 theme full-length 658:668 arg1 protein 670:676 the full-length protein 654:676 the full-length protein 654:676 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 2 35 theme tumor 151:155 arg1 TNF-beta 179:186 TNF-beta 179:186 TNF-beta 179:186 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 35 theme tumor 151:155 arg1 beta 173:176 Natural human tumor necrosis factor beta 137:176 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line 137:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 5 36 link O-linked 770:777 arg1 carbohydrates 779:791 O-linked carbohydrates 770:791 O-linked carbohydrates 770:791 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 5 37 attach derived 641:647 arg2 forms 631:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms 601:635 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 5 37 attach derived 641:647 arg1 protein 670:676 the full-length protein 654:676 the full-length protein 654:676 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 2 38 theme line 249:252 arg1 supernatants 203:214 supernatants 203:214 supernatants of a human B-lymphoblastoid cell line 203:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 5 39 theme trypsin-like 681:692 arg1 cleavage 706:713 trypsin-like proteolytic cleavage 681:713 trypsin-like proteolytic cleavage in the N-proximal region 681:738 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 2 40 theme cell 244:247 arg1 line 249:252 a human B-lymphoblastoid cell line 219:252 a human B-lymphoblastoid cell line 219:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 1 41 theme allelic 118:124 arg1 variation 126:134 allelic variation 118:134 allelic variation 118:134 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 5 42 theme clipped 747:753 arg1 molecules 755:763 these clipped molecules 741:763 these clipped molecules 741:763 The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. 1451807 6 43 theme acid 834:837 arg1 position 839:846 amino acid position 26 828:849 amino acid position 26 (threonine/asparagine) 828:872 Two allelic variants differing in amino acid position 26 (threonine/asparagine) were identified. 1451807 6 43 theme acid 834:837 arg1 threonine/asparagine 852:871 threonine/asparagine 852:871 threonine/asparagine 852:871 Two allelic variants differing in amino acid position 26 (threonine/asparagine) were identified. 1451807 2 44 theme B-lymphoblastoid 227:242 arg1 line 249:252 a human B-lymphoblastoid cell line 219:252 a human B-lymphoblastoid cell line 219:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 1 45 from Thr7 84:87 arg1 processing 102:111 proteolytic processing 90:111 proteolytic processing 90:111 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 1 45 from Thr7 84:87 arg1 O-glycosylation 65:79 Variable O-glycosylation 56:79 Variable O-glycosylation at Thr7 56:87 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 1 45 from Thr7 84:87 arg1 variation 126:134 allelic variation 118:134 allelic variation 118:134 Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. 1451807 4 46 theme molecules 481:489 arg1 part 469:472 part 469:472 part of the molecules 469:489 In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. 1451807 2 47 theme human 221:225 arg1 line 249:252 a human B-lymphoblastoid cell line 219:252 a human B-lymphoblastoid cell line 219:252 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1451807 2 48 theme gel 337:339 arg1 electrophoresis 341:355 gel electrophoresis 337:355 gel electrophoresis 337:355 Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. 1517205 7 0 theme peptide 1509:1515 arg1 N-3 1517:1519 peptide N-3 1509:1519 peptide N-3 1509:1519 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 6 1 contain contain 1377:1383 arg2 Xyl 1385:1387 Xyl 1385:1387 Xyl 1385:1387 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 6 1 contain contain 1377:1383 arg2 Glc 1393:1395 Glc 1393:1395 Glc 1393:1395 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 6 1 contain contain 1377:1383 arg1 glycopeptides 1287:1299 These glycopeptides 1281:1299 These glycopeptides 1281:1299 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 9 2 theme 1 1833:1833 arg1 mol 1835:1837 mol 1835:1837 mol 1835:1837 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 4 3 theme acid 886:889 arg1 analysis 891:898 sialic acid analysis 879:898 sialic acid analysis of the isolated peptides 879:923 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 2 4 from evidence 433:440 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 4 from evidence 433:440 arg1 domain 488:493 the first EGF-like domain 469:493 the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 469:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 7 5 theme mass 1478:1481 arg1 analysis 1497:1504 mass spectrometric analysis 1478:1504 the fast atom bombardment mass spectrometric analysis 1452:1504 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 7 6 from data 1412:1415 arg1 beta-elimination 1420:1435 beta-elimination 1420:1435 beta-elimination for N-9 1420:1443 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 9 7 theme Gal 1847:1849 arg1 mol 1835:1837 mol 1835:1837 mol 1835:1837 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 2 8 theme first 473:477 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 8 theme first 473:477 arg1 domain 488:493 the first EGF-like domain 469:493 the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 469:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 7 9 theme atom 1461:1464 arg1 bombardment 1466:1476 the fast atom bombardment 1452:1476 the fast atom bombardment mass spectrometric analysis 1452:1504 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 7 10 from beta-elimination 1420:1435 arg1 N-3 1517:1519 peptide N-3 1509:1519 peptide N-3 1509:1519 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 4 11 theme each 961:964 arg1 mol 957:959 1 mol each 955:964 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc) 955:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 9 12 theme Fuc 1852:1854 arg1 mol 1835:1837 mol 1835:1837 mol 1835:1837 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 1 13 theme protein 341:347 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 13 theme protein 341:347 arg1 Ser-53 352:357 Ser-53 352:357 Ser-53 352:357 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 13 theme protein 341:347 arg1 Z 349:349 protein Z 341:349 protein Z (Ser-53) 341:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 5 14 dep positions 1234:1242 arg1 55-63 1244:1248 55-63 1244:1248 55-63 1244:1248 hIX-GP1 was further digested with asparaginyl endopeptidase, and two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), were isolated, respectively. 1517205 9 15 dep mol 1835:1837 arg1 each 1839:1842 each 1839:1842 each 1839:1842 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 0 16 attach linked 55:60 arg2 tetrasaccharide 22:36 a tetrasaccharide 20:36 a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue 20:100 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 0 16 attach linked 55:60 arg1 serine 65:70 serine 61 65:73 serine 61 65:73 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 5 17 gly glycopeptides 1164:1176 arg2 glycopeptides 1164:1176 two glycopeptides 1160:1176 two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), 1160:1250 hIX-GP1 was further digested with asparaginyl endopeptidase, and two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), were isolated, respectively. 1517205 1 18 theme bovine 286:291 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 18 theme bovine 286:291 arg1 IX 324:325 IX 324:325 IX (Ser-53) 324:334 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 18 theme bovine 286:291 arg1 VII 310:312 VII (Ser-52) 310:321 VII (Ser-52) 310:321 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 18 theme bovine 286:291 arg1 Z 349:349 protein Z 341:349 protein Z (Ser-53) 341:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 19 theme factors 302:308 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 19 theme factors 302:308 arg1 domains 265:271 the epidermal growth factor (EGF)-like domains 226:271 the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 226:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 6 20 theme Gal 1337:1339 arg1 mol 1325:1327 1 mol 1323:1327 1 mol each of Gal, Fuc, GlcNAc, and NeuAc 1323:1363 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 2 21 theme tetrasaccharide 536:550 arg1 O-fucosidically 552:566 a tetrasaccharide O-fucosidically 534:566 a tetrasaccharide O-fucosidically linked to Ser-61 534:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 21 theme tetrasaccharide 536:550 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 6 22 gly glycopeptides 1287:1299 arg2 glycopeptides 1287:1299 These glycopeptides 1281:1299 These glycopeptides 1281:1299 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 5 23 dep positions 1208:1216 arg1 59-63 1218:1222 59-63 1218:1222 59-63 1218:1222 hIX-GP1 was further digested with asparaginyl endopeptidase, and two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), were isolated, respectively. 1517205 8 24 attach released 1621:1628 arg1 hIX-GP1 1635:1641 hIX-GP1 1635:1641 hIX-GP1 1635:1641 An analysis of the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination revealed that the reducing end was PA-Fuc. 1517205 8 24 attach released 1621:1628 arg2 PA-oligosaccharide 1602:1619 the PA-oligosaccharide 1598:1619 the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination 1598:1688 An analysis of the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination revealed that the reducing end was PA-Fuc. 1517205 3 25 theme named 642:646 arg1 peptides 596:603 Two large peptides 586:603 Two large peptides containing Ser-61 (positions 44-63) 586:639 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 25 theme named 642:646 arg1 hIX-GP1 648:654 named hIX-GP1 642:654 named hIX-GP1 642:654 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 9 26 contain has 1791:1793 arg2 tetrasaccharide 1803:1817 a novel tetrasaccharide 1795:1817 a novel tetrasaccharide 1795:1817 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 9 26 contain has 1791:1793 arg1 IX 1788:1789 human factor IX 1775:1789 human factor IX 1775:1789 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 4 27 theme galactose 969:977 arg1 mol 957:959 1 mol each 955:964 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc) 955:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 1 28 attach linked 196:201 arg1 residue 215:221 a serine residue 206:221 a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 206:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 28 attach linked 196:201 arg2 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 28 attach linked 196:201 arg2 2-Glc 189:193 (Xyl)2-Glc 184:193 (Xyl)2-Glc 184:193 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 28 attach linked 196:201 arg2 -glucose 165:172 xylose (Xyl)-glucose 153:172 xylose (Xyl)-glucose (Glc) 153:178 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 29 theme sugar 139:143 arg1 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 29 theme sugar 139:143 arg1 -glucose 165:172 xylose (Xyl)-glucose 153:172 xylose (Xyl)-glucose (Glc) 153:178 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 29 theme sugar 139:143 arg1 2-Glc 189:193 (Xyl)2-Glc 184:193 (Xyl)2-Glc 184:193 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 3 30 theme lysyl 698:702 arg1 endopeptidase-digest 704:723 the lysyl endopeptidase-digest 694:723 the lysyl endopeptidase-digest of human factor IX 694:742 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 2 31 theme factor 504:509 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 31 theme factor 504:509 arg1 O-fucosidically 552:566 a tetrasaccharide O-fucosidically 534:566 a tetrasaccharide O-fucosidically linked to Ser-61 534:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 9 32 attach linked 1902:1907 arg2 tetrasaccharide 1803:1817 a novel tetrasaccharide 1795:1817 a novel tetrasaccharide 1795:1817 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 9 32 attach linked 1902:1907 arg1 Ser-61 1912:1917 Ser-61 1912:1917 Ser-61 1912:1917 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 1 33 theme glycoprotein 392:403 arg1 thrombospondin 405:418 bovine platelet glycoprotein thrombospondin 376:418 bovine platelet glycoprotein thrombospondin 376:418 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 34 theme human 276:280 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 34 theme human 276:280 arg1 IX 324:325 IX 324:325 IX (Ser-53) 324:334 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 34 theme human 276:280 arg1 VII 310:312 VII (Ser-52) 310:321 VII (Ser-52) 310:321 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 34 theme human 276:280 arg1 Z 349:349 protein Z 341:349 protein Z (Ser-53) 341:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 6 35 theme GlcNAc 1347:1352 arg1 mol 1325:1327 1 mol 1323:1327 1 mol each of Gal, Fuc, GlcNAc, and NeuAc 1323:1363 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 3 36 theme factor 734:739 arg1 IX 741:742 human factor IX 728:742 human factor IX 728:742 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 1 37 dep chains 145:150 arg1 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 37 dep chains 145:150 arg1 Glc 175:177 Glc 175:177 Glc 175:177 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 37 dep chains 145:150 arg1 -glucose 165:172 xylose (Xyl)-glucose 153:172 xylose (Xyl)-glucose (Glc) 153:178 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 37 dep chains 145:150 arg1 2-Glc 189:193 (Xyl)2-Glc 184:193 (Xyl)2-Glc 184:193 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 0 38 theme Human 0:4 arg1 IX 13:14 Human factor IX 0:14 Human factor IX 0:14 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 6 39 theme Fuc 1342:1344 arg1 mol 1325:1327 1 mol 1323:1327 1 mol each of Gal, Fuc, GlcNAc, and NeuAc 1323:1363 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 3 40 theme performance 768:778 arg1 HPLC 803:806 HPLC 803:806 HPLC 803:806 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 40 theme performance 768:778 arg1 chromatography 787:800 reversed-phase high performance liquid chromatography 748:800 reversed-phase high performance liquid chromatography (HPLC) 748:807 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 4 41 theme N-acetylglucosamine 1000:1018 arg1 mol 957:959 1 mol each 955:964 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc) 955:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 3 42 contain containing 605:614 arg1 peptides 596:603 Two large peptides 586:603 Two large peptides containing Ser-61 (positions 44-63) 586:639 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 42 contain containing 605:614 arg1 hIX-GP2 660:666 hIX-GP2 660:666 hIX-GP2 660:666 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 42 contain containing 605:614 arg1 hIX-GP1 648:654 named hIX-GP1 642:654 named hIX-GP1 642:654 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 42 contain containing 605:614 arg2 positions 624:632 positions 44-63 624:638 positions 44-63 624:638 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 42 contain containing 605:614 arg2 Ser-61 616:621 Ser-61 616:621 Ser-61 (positions 44-63) 616:639 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 4 43 contain contained 945:953 arg1 they 940:943 they 940:943 they 940:943 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 4 43 contain contained 945:953 arg2 mol 957:959 1 mol each 955:964 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc) 955:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 3 44 theme reversed-phase 748:761 arg1 HPLC 803:806 HPLC 803:806 HPLC 803:806 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 44 theme reversed-phase 748:761 arg1 chromatography 787:800 reversed-phase high performance liquid chromatography 748:800 reversed-phase high performance liquid chromatography (HPLC) 748:807 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 4 45 theme sugar 832:836 arg1 analysis 838:845 the component sugar analysis 818:845 the component sugar analysis after pyridylamination (PA) 818:873 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 2 46 from modification 453:464 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 46 from modification 453:464 arg1 domain 488:493 the first EGF-like domain 469:493 the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 469:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 9 47 theme human 1775:1779 arg1 IX 1788:1789 human factor IX 1775:1789 human factor IX 1775:1789 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 1 48 theme growth 240:245 arg1 EGF 255:257 EGF 255:257 EGF 255:257 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 48 theme growth 240:245 arg1 factor 247:252 the epidermal growth factor 226:252 the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 226:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 7 49 dep bombardment 1466:1476 arg1 analysis 1497:1504 mass spectrometric analysis 1478:1504 the fast atom bombardment mass spectrometric analysis 1452:1504 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 4 50 theme N-acetylneuraminic 1034:1051 arg1 NeuAc 1059:1063 NeuAc 1059:1063 NeuAc 1059:1063 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 4 50 theme N-acetylneuraminic 1034:1051 arg1 acid 1053:1056 N-acetylneuraminic acid 1034:1056 N-acetylneuraminic acid (NeuAc) 1034:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 4 51 theme peptides 916:923 arg1 analysis 838:845 the component sugar analysis 818:845 the component sugar analysis after pyridylamination (PA) 818:873 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 4 51 theme peptides 916:923 arg1 analysis 891:898 sialic acid analysis 879:898 sialic acid analysis of the isolated peptides 879:923 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 4 52 from Data 810:813 arg1 analysis 838:845 the component sugar analysis 818:845 the component sugar analysis after pyridylamination (PA) 818:873 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 4 52 from Data 810:813 arg1 analysis 891:898 sialic acid analysis 879:898 sialic acid analysis of the isolated peptides 879:923 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 1 53 gly glycoprotein 392:403 arg1 glycoprotein 392:403 bovine platelet glycoprotein thrombospondin 376:418 bovine platelet glycoprotein thrombospondin 376:418 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 4 54 theme sialic 879:884 arg1 analysis 891:898 sialic acid analysis 879:898 sialic acid analysis of the isolated peptides 879:923 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 1 55 theme bovine 376:381 arg1 thrombospondin 405:418 bovine platelet glycoprotein thrombospondin 376:418 bovine platelet glycoprotein thrombospondin 376:418 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 2 56 theme modification 453:464 arg1 evidence 433:440 evidence 433:440 evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 433:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 7 57 theme spectrometric 1483:1495 arg1 analysis 1497:1504 mass spectrometric analysis 1478:1504 the fast atom bombardment mass spectrometric analysis 1452:1504 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 7 58 theme bombardment 1466:1476 arg1 data 1412:1415 the data 1408:1415 the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 1408:1519 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 1 59 from residue 215:221 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 59 from residue 215:221 arg1 domains 265:271 the epidermal growth factor (EGF)-like domains 226:271 the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 226:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 6 60 dep mol 1325:1327 arg1 each 1329:1332 each 1329:1332 each 1329:1332 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 1 61 theme xylose 153:158 arg1 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 61 theme xylose 153:158 arg1 Glc 175:177 Glc 175:177 Glc 175:177 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 61 theme xylose 153:158 arg1 -glucose 165:172 xylose (Xyl)-glucose 153:172 xylose (Xyl)-glucose (Glc) 153:178 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 2 62 theme EGF-like 479:486 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 62 theme EGF-like 479:486 arg1 domain 488:493 the first EGF-like domain 469:493 the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 469:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 7 63 theme fast 1456:1459 arg1 bombardment 1466:1476 the fast atom bombardment 1452:1476 the fast atom bombardment mass spectrometric analysis 1452:1504 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 0 64 contain has 16:18 arg1 IX 13:14 Human factor IX 0:14 Human factor IX 0:14 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 0 64 contain has 16:18 arg2 tetrasaccharide 22:36 a tetrasaccharide 20:36 a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue 20:100 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 1 65 theme Xyl 161:163 arg1 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 65 theme Xyl 161:163 arg1 Glc 175:177 Glc 175:177 Glc 175:177 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 65 theme Xyl 161:163 arg1 -glucose 165:172 xylose (Xyl)-glucose 153:172 xylose (Xyl)-glucose (Glc) 153:178 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 8 66 theme PA-oligosaccharide 1602:1619 arg1 analysis 1586:1593 An analysis 1583:1593 An analysis of the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination 1583:1688 An analysis of the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination revealed that the reducing end was PA-Fuc. 1517205 4 67 dep Glc 1082:1084 arg1 addition 1070:1077 addition 1070:1077 addition 1070:1077 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 3 68 theme large 590:594 arg1 peptides 596:603 Two large peptides 586:603 Two large peptides containing Ser-61 (positions 44-63) 586:639 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 68 theme large 590:594 arg1 hIX-GP2 660:666 hIX-GP2 660:666 hIX-GP2 660:666 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 68 theme large 590:594 arg1 hIX-GP1 648:654 named hIX-GP1 642:654 named hIX-GP1 642:654 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 1 69 theme clotting 293:300 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 69 theme clotting 293:300 arg1 IX 324:325 IX 324:325 IX (Ser-53) 324:334 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 69 theme clotting 293:300 arg1 VII 310:312 VII (Ser-52) 310:321 VII (Ser-52) 310:321 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 69 theme clotting 293:300 arg1 Z 349:349 protein Z 341:349 protein Z (Ser-53) 341:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 4 70 theme fucose 986:991 arg1 mol 957:959 1 mol each 955:964 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc) 955:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 8 71 theme reducing 1708:1715 arg1 PA-Fuc 1725:1730 PA-Fuc 1725:1730 PA-Fuc 1725:1730 An analysis of the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination revealed that the reducing end was PA-Fuc. 1517205 8 71 theme reducing 1708:1715 arg1 end 1717:1719 the reducing end 1704:1719 the reducing end 1704:1719 An analysis of the PA-oligosaccharide released from hIX-GP1 by hydrazinolysis followed by pyridylamination revealed that the reducing end was PA-Fuc. 1517205 3 72 attach isolated 680:687 arg1 endopeptidase-digest 704:723 the lysyl endopeptidase-digest 694:723 the lysyl endopeptidase-digest of human factor IX 694:742 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 72 attach isolated 680:687 arg2 hIX-GP2 660:666 hIX-GP2 660:666 hIX-GP2 660:666 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 72 attach isolated 680:687 arg2 hIX-GP1 648:654 named hIX-GP1 642:654 named hIX-GP1 642:654 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 72 attach isolated 680:687 arg2 peptides 596:603 Two large peptides 586:603 Two large peptides containing Ser-61 (positions 44-63) 586:639 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 1 73 theme unusual 131:137 arg1 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 73 theme unusual 131:137 arg1 -glucose 165:172 xylose (Xyl)-glucose 153:172 xylose (Xyl)-glucose (Glc) 153:178 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 73 theme unusual 131:137 arg1 2-Glc 189:193 (Xyl)2-Glc 184:193 (Xyl)2-Glc 184:193 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 2 74 attach linked 568:573 arg2 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 74 attach linked 568:573 arg1 Ser-61 578:583 Ser-61 578:583 Ser-61 578:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 74 attach linked 568:573 arg2 O-fucosidically 552:566 a tetrasaccharide O-fucosidically 534:566 a tetrasaccharide O-fucosidically linked to Ser-61 534:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 9 75 theme GlcNAc 1857:1862 arg1 mol 1835:1837 mol 1835:1837 mol 1835:1837 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 3 76 theme human 728:732 arg1 IX 741:742 human factor IX 728:742 human factor IX 728:742 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 2 77 theme IX 511:512 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 77 theme IX 511:512 arg1 domain 488:493 the first EGF-like domain 469:493 the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 469:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 0 78 theme fucose 87:92 arg1 residue 94:100 the fucose residue 83:100 the fucose residue 83:100 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 1 79 theme -like 259:263 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 79 theme -like 259:263 arg1 domains 265:271 the epidermal growth factor (EGF)-like domains 226:271 the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 226:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 6 80 theme NeuAc 1359:1363 arg1 mol 1325:1327 1 mol 1323:1327 1 mol each of Gal, Fuc, GlcNAc, and NeuAc 1323:1363 These glycopeptides were both composed of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc but did not contain Xyl and Glc. 1517205 2 81 theme human 498:502 arg1 IX 511:512 human factor IX 498:512 human factor IX 498:512 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 81 theme human 498:502 arg1 O-fucosidically 552:566 a tetrasaccharide O-fucosidically 534:566 a tetrasaccharide O-fucosidically linked to Ser-61 534:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 1 82 theme platelet 383:390 arg1 thrombospondin 405:418 bovine platelet glycoprotein thrombospondin 376:418 bovine platelet glycoprotein thrombospondin 376:418 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 9 83 theme Fuc 1931:1933 arg1 residue 1935:1941 the Fuc residue 1927:1941 the Fuc residue 1927:1941 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 7 84 attach linked 1565:1570 arg2 tetrasaccharide 1549:1563 a tetrasaccharide 1547:1563 a tetrasaccharide linked to Ser-61 1547:1580 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 7 84 attach linked 1565:1570 arg1 Ser-61 1575:1580 Ser-61 1575:1580 Ser-61 1575:1580 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 0 85 theme factor 6:11 arg1 IX 13:14 Human factor IX 0:14 Human factor IX 0:14 Human factor IX has a tetrasaccharide O-glycosidically linked to serine 61 through the fucose residue. 1517205 3 86 theme IX 741:742 arg1 endopeptidase-digest 704:723 the lysyl endopeptidase-digest 694:723 the lysyl endopeptidase-digest of human factor IX 694:742 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 1 87 theme serine 208:213 arg1 residue 215:221 a serine residue 206:221 a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 206:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 3 88 theme liquid 780:785 arg1 HPLC 803:806 HPLC 803:806 HPLC 803:806 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 88 theme liquid 780:785 arg1 chromatography 787:800 reversed-phase high performance liquid chromatography 748:800 reversed-phase high performance liquid chromatography (HPLC) 748:807 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 2 89 contain have 428:431 arg2 evidence 433:440 evidence 433:440 evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 433:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 2 89 contain have 428:431 arg1 We 421:422 We 421:422 We 421:422 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 3 90 theme high 763:766 arg1 HPLC 803:806 HPLC 803:806 HPLC 803:806 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 3 90 theme high 763:766 arg1 chromatography 787:800 reversed-phase high performance liquid chromatography 748:800 reversed-phase high performance liquid chromatography (HPLC) 748:807 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 4 91 theme component 822:830 arg1 analysis 838:845 the component sugar analysis 818:845 the component sugar analysis after pyridylamination (PA) 818:873 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 9 92 theme factor 1781:1786 arg1 IX 1788:1789 human factor IX 1775:1789 human factor IX 1775:1789 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 1 93 theme Xyl 185:187 arg1 chains 145:150 unusual sugar chains 131:150 unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 131:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 93 theme Xyl 185:187 arg1 2-Glc 189:193 (Xyl)2-Glc 184:193 (Xyl)2-Glc 184:193 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 5 94 contain containing 1178:1187 arg2 Ser-61 1189:1194 Ser-61 1189:1194 Ser-61 1189:1194 hIX-GP1 was further digested with asparaginyl endopeptidase, and two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), were isolated, respectively. 1517205 5 94 contain containing 1178:1187 arg1 glycopeptides 1164:1176 two glycopeptides 1160:1176 two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), 1160:1250 hIX-GP1 was further digested with asparaginyl endopeptidase, and two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), were isolated, respectively. 1517205 1 95 theme epidermal 230:238 arg1 EGF 255:257 EGF 255:257 EGF 255:257 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 95 theme epidermal 230:238 arg1 factor 247:252 the epidermal growth factor 226:252 the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 226:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 3 96 dep positions 624:632 arg1 44-63 634:638 44-63 634:638 44-63 634:638 Two large peptides containing Ser-61 (positions 44-63), named hIX-GP1 and hIX-GP2, were first isolated from the lysyl endopeptidase-digest of human factor IX, by reversed-phase high performance liquid chromatography (HPLC). 1517205 2 97 from domain 488:493 arg1 evidence 433:440 evidence 433:440 evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61 433:583 We now have evidence of another modification in the first EGF-like domain of human factor IX, which proved to be a tetrasaccharide O-fucosidically linked to Ser-61. 1517205 4 98 theme acid 1053:1056 arg1 mol 957:959 1 mol each 955:964 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc) 955:1064 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 1 99 theme factor 247:252 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 99 theme factor 247:252 arg1 domains 265:271 the epidermal growth factor (EGF)-like domains 226:271 the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 226:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 7 100 theme tetrasaccharide 1549:1563 arg1 presence 1535:1542 the presence 1531:1542 the presence of a tetrasaccharide linked to Ser-61 1531:1580 Moreover, the data on beta-elimination for N-9 and of the fast atom bombardment mass spectrometric analysis on peptide N-3 suggested the presence of a tetrasaccharide linked to Ser-61. 1517205 1 101 dep factors 302:308 arg1 factors 302:308 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53) 276:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 101 dep factors 302:308 arg1 IX 324:325 IX 324:325 IX (Ser-53) 324:334 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 101 dep factors 302:308 arg1 VII 310:312 VII (Ser-52) 310:321 VII (Ser-52) 310:321 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 101 dep factors 302:308 arg1 Ser-53 328:333 Ser-53 328:333 Ser-53 328:333 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 101 dep factors 302:308 arg1 Ser-53 352:357 Ser-53 352:357 Ser-53 352:357 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 1 101 dep factors 302:308 arg1 Z 349:349 protein Z 341:349 protein Z (Ser-53) 341:358 We have recently discovered unusual sugar chains (xylose (Xyl)-glucose (Glc) and (Xyl)2-Glc) linked to a serine residue in the epidermal growth factor (EGF)-like domains of human and bovine clotting factors VII (Ser-52), IX (Ser-53), and protein Z (Ser-53), in addition to bovine platelet glycoprotein thrombospondin. 1517205 9 102 theme novel 1797:1801 arg1 tetrasaccharide 1803:1817 a novel tetrasaccharide 1795:1817 a novel tetrasaccharide 1795:1817 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 1517205 4 103 theme isolated 907:914 arg1 peptides 916:923 the isolated peptides 903:923 the isolated peptides 903:923 Data on the component sugar analysis after pyridylamination (PA) and sialic acid analysis of the isolated peptides indicated that they contained 1 mol each of galactose (Gal), fucose (Fuc), N-acetylglucosamine (GlcNAc), and N-acetylneuraminic acid (NeuAc), in addition to Glc and Xyl. 1517205 5 104 theme asparaginyl 1129:1139 arg1 endopeptidase 1141:1153 asparaginyl endopeptidase 1129:1153 asparaginyl endopeptidase 1129:1153 hIX-GP1 was further digested with asparaginyl endopeptidase, and two glycopeptides containing Ser-61, named N-3 (positions 59-63) and N-9 (positions 55-63), were isolated, respectively. 1517205 9 105 theme NeuAc 1869:1873 arg1 mol 1835:1837 mol 1835:1837 mol 1835:1837 All the results support the proposal that human factor IX has a novel tetrasaccharide consisting of 1 mol each of Gal, Fuc, GlcNAc, and NeuAc, which is O-glycosidically linked to Ser-61 through the Fuc residue. 9578468 10 0 theme natural 1733:1739 arg1 responses 1759:1767 natural eotaxin-dependent responses 1733:1767 natural eotaxin-dependent responses 1733:1767 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 3 1 with protein 604:610 arg1 mass 639:642 an apparent molecular mass 617:642 an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis 617:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 9 2 theme eotaxin 1334:1340 arg1 preparations 1342:1353 natural eotaxin preparations 1326:1353 natural eotaxin preparations 1326:1353 Neither neutrophils, monocytes or lymphocytes responded towards natural eotaxin preparations indicating that N-terminal truncation and O-glycosylation did not affect the cell-specificity of chemotactic activity. 9578468 4 3 theme Peptide 697:703 arg1 mapping 705:711 Peptide mapping 697:711 Peptide mapping with subsequent amino acid sequence analyses 697:756 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 3 4 theme heparin-binding 588:602 arg1 protein 604:610 a heparin-binding protein 586:610 a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis 586:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 10 5 dep T-lymphocyte 1592:1603 arg1 secreted 1619:1626 secreted 1619:1626 secreted 1619:1626 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 10 5 dep T-lymphocyte 1592:1603 arg1 expressed 1605:1613 expressed 1605:1613 expressed 1605:1613 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 9 6 theme N-terminal 1371:1380 arg1 truncation 1382:1391 N-terminal truncation 1371:1391 N-terminal truncation 1371:1391 Neither neutrophils, monocytes or lymphocytes responded towards natural eotaxin preparations indicating that N-terminal truncation and O-glycosylation did not affect the cell-specificity of chemotactic activity. 9578468 3 7 theme molecular 629:637 arg1 mass 639:642 an apparent molecular mass 617:642 an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis 617:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 1 8 theme selective 208:216 arg1 infiltration 229:240 selective eosinophil infiltration 208:240 selective eosinophil infiltration 208:240 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 4 9 theme human 776:780 arg1 eotaxin 782:788 human eotaxin 776:788 human eotaxin 776:788 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 3 10 theme SDS/polyacrylamide 657:674 arg1 electrophoresis 680:694 SDS/polyacrylamide gel electrophoresis 657:694 SDS/polyacrylamide gel electrophoresis 657:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 4 11 theme sequence 740:747 arg1 analyses 749:756 subsequent amino acid sequence analyses 718:756 subsequent amino acid sequence analyses 718:756 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 3 12 theme kDa 650:652 arg1 mass 639:642 an apparent molecular mass 617:642 an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis 617:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 5 13 theme % 830:830 arg1 forms 855:859 50% N-terminally truncated forms 828:859 50% N-terminally truncated forms missing two or three amino acids 828:892 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 7 14 theme Da 1143:1144 arg1 masses 1116:1121 principal masses 1106:1121 principal masses of 9033 Da and 9317 Da 1106:1144 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 9 15 gly O-glycosylation 1397:1411 arg1 activity 1464:1471 chemotactic activity 1452:1471 chemotactic activity 1452:1471 Neither neutrophils, monocytes or lymphocytes responded towards natural eotaxin preparations indicating that N-terminal truncation and O-glycosylation did not affect the cell-specificity of chemotactic activity. 9578468 7 16 with heterogeneous 1087:1099 arg1 masses 1116:1121 principal masses 1106:1121 principal masses of 9033 Da and 9317 Da 1106:1144 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 4 17 with mapping 705:711 arg1 analyses 749:756 subsequent amino acid sequence analyses 718:756 subsequent amino acid sequence analyses 718:756 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 5 18 theme eotaxin 799:805 arg1 preparations 807:818 Natural eotaxin preparations 791:818 Natural eotaxin preparations 791:818 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 9 19 theme chemotactic 1452:1462 arg1 activity 1464:1471 chemotactic activity 1452:1471 chemotactic activity 1452:1471 Neither neutrophils, monocytes or lymphocytes responded towards natural eotaxin preparations indicating that N-terminal truncation and O-glycosylation did not affect the cell-specificity of chemotactic activity. 9578468 1 20 theme inflammatory 160:171 arg1 diseases 178:185 inflammatory skin diseases 160:185 inflammatory skin diseases 160:185 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 0 21 theme human 66:70 arg1 eotaxin 72:78 human eotaxin 66:78 human eotaxin 66:78 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 1 22 theme diseases 178:185 arg1 number 150:155 a number 148:155 a number of inflammatory skin diseases 148:185 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 5 23 theme amino 882:886 arg1 acids 888:892 two or three amino acids 869:892 two or three amino acids 869:892 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 8 24 theme Natural 1147:1153 arg1 eotaxin 1155:1161 Natural eotaxin 1147:1161 Natural eotaxin 1147:1161 Natural eotaxin stimulated eosinophil chemotaxis with identical potency and efficacy as recombinant human eotaxin. 9578468 10 25 theme chemotactic 1650:1660 arg1 protein 1662:1668 monocyte chemotactic protein 3 1641:1670 monocyte chemotactic protein 3 (MCP-3) 1641:1678 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 10 25 theme chemotactic 1650:1660 arg1 MCP-3 1673:1677 MCP-3 1673:1677 MCP-3 1673:1677 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 0 26 theme dermal 122:127 arg1 fibroblasts 129:139 tumor-necrosis-factor-alpha-stimulated dermal fibroblasts 83:139 tumor-necrosis-factor-alpha-stimulated dermal fibroblasts 83:139 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 2 27 theme chemotactic 525:535 arg1 activity 537:544 eosinophil-specific chemotactic activity 505:544 eosinophil-specific chemotactic activity 505:544 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 0 28 theme Delayed 0:6 arg1 production 8:17 Delayed production 0:17 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts 0:139 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 8 29 theme identical 1201:1209 arg1 potency 1211:1217 identical potency 1201:1217 identical potency 1201:1217 Natural eotaxin stimulated eosinophil chemotaxis with identical potency and efficacy as recombinant human eotaxin. 9578468 7 30 theme natural 1053:1059 arg1 preparation 1069:1079 the natural eotaxin preparation 1049:1079 the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da 1049:1144 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 6 31 from Thr71 919:923 arg1 O-glycosylated 901:914 O-glycosylated 901:914 O-glycosylated 901:914 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 1 32 theme cellular 338:345 arg1 source 347:352 a potential cellular source 326:352 a potential cellular source of eosinophil-selective attractants 326:388 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 6 33 gly sialylated 952:961 arg1 variants 978:985 at least two sialylated O-glycosylated variants 939:985 at least two sialylated O-glycosylated variants 939:985 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 0 34 theme active 35:40 arg1 forms 57:61 biologically active O-glycosylated forms 22:61 biologically active O-glycosylated forms of human eotaxin 22:78 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 6 35 theme sialylated 952:961 arg1 variants 978:985 at least two sialylated O-glycosylated variants 939:985 at least two sialylated O-glycosylated variants 939:985 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 0 36 theme forms 57:61 arg1 production 8:17 Delayed production 0:17 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts 0:139 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 1 37 theme attractants 378:388 arg1 source 347:352 a potential cellular source 326:352 a potential cellular source of eosinophil-selective attractants 326:388 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 8 38 theme human 1247:1251 arg1 eotaxin 1253:1259 recombinant human eotaxin 1235:1259 recombinant human eotaxin 1235:1259 Natural eotaxin stimulated eosinophil chemotaxis with identical potency and efficacy as recombinant human eotaxin. 9578468 10 39 theme chemotactic 1533:1543 arg1 responses 1545:1553 chemotactic responses 1533:1553 chemotactic responses 1533:1553 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 2 40 theme dermal 402:407 arg1 fibroblasts 409:419 Cultivated dermal fibroblasts 391:419 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha 391:460 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 0 41 gly O-glycosylated 42:55 arg1 forms 57:61 biologically active O-glycosylated forms 22:61 biologically active O-glycosylated forms of human eotaxin 22:78 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 0 41 gly O-glycosylated 42:55 arg1 eotaxin 72:78 human eotaxin 66:78 human eotaxin 66:78 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 2 42 theme factor 449:454 arg1 alpha 456:460 tumor necrosis factor alpha 434:460 tumor necrosis factor alpha 434:460 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 4 43 theme subsequent 718:727 arg1 analyses 749:756 subsequent amino acid sequence analyses 718:756 subsequent amino acid sequence analyses 718:756 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 10 44 theme eotaxin-dependent 1741:1757 arg1 responses 1759:1767 natural eotaxin-dependent responses 1733:1767 natural eotaxin-dependent responses 1733:1767 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 9 45 theme natural 1326:1332 arg1 preparations 1342:1353 natural eotaxin preparations 1326:1353 natural eotaxin preparations 1326:1353 Neither neutrophils, monocytes or lymphocytes responded towards natural eotaxin preparations indicating that N-terminal truncation and O-glycosylation did not affect the cell-specificity of chemotactic activity. 9578468 2 46 theme tumor 434:438 arg1 alpha 456:460 tumor necrosis factor alpha 434:460 tumor necrosis factor alpha 434:460 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 7 47 theme mass 1012:1015 arg1 analyses 1031:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 1 48 theme dermal 291:296 arg1 fibroblasts 298:308 dermal fibroblasts 291:308 dermal fibroblasts 291:308 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 7 49 theme Electrospray 988:999 arg1 analyses 1031:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 3 50 theme apparent 620:627 arg1 mass 639:642 an apparent molecular mass 617:642 an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis 617:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 1 51 theme eosinophil 218:227 arg1 infiltration 229:240 selective eosinophil infiltration 208:240 selective eosinophil infiltration 208:240 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 4 52 theme acid 735:738 arg1 analyses 749:756 subsequent amino acid sequence analyses 718:756 subsequent amino acid sequence analyses 718:756 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 5 53 theme 50 828:829 arg1 % 830:830 % 830:830 % 830:830 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 3 54 theme gel 676:678 arg1 electrophoresis 680:694 SDS/polyacrylamide gel electrophoresis 657:694 SDS/polyacrylamide gel electrophoresis 657:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 6 55 gly O-glycosylated 901:914 arg1 It 895:896 It 895:896 It 895:896 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 6 55 gly O-glycosylated 901:914 arg2 Thr71 919:923 Thr71 919:923 Thr71 919:923 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 5 56 theme Natural 791:797 arg1 preparations 807:818 Natural eotaxin preparations 791:818 Natural eotaxin preparations 791:818 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 0 57 theme eotaxin 72:78 arg1 forms 57:61 biologically active O-glycosylated forms 22:61 biologically active O-glycosylated forms of human eotaxin 22:78 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 9 58 theme activity 1464:1471 arg1 cell-specificity 1432:1447 the cell-specificity 1428:1447 the cell-specificity of chemotactic activity 1428:1471 Neither neutrophils, monocytes or lymphocytes responded towards natural eotaxin preparations indicating that N-terminal truncation and O-glycosylation did not affect the cell-specificity of chemotactic activity. 9578468 7 59 theme Da 1131:1132 arg1 masses 1116:1121 principal masses 1106:1121 principal masses of 9033 Da and 9317 Da 1106:1144 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 1 60 theme skin 173:176 arg1 diseases 178:185 inflammatory skin diseases 160:185 inflammatory skin diseases 160:185 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 5 61 theme truncated 845:853 arg1 forms 855:859 50% N-terminally truncated forms 828:859 50% N-terminally truncated forms missing two or three amino acids 828:892 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 0 62 theme tumor-necrosis-factor-alpha-stimulated 83:120 arg1 fibroblasts 129:139 tumor-necrosis-factor-alpha-stimulated dermal fibroblasts 83:139 tumor-necrosis-factor-alpha-stimulated dermal fibroblasts 83:139 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 10 63 with Treatment 1474:1482 arg1 eotaxin 1512:1518 natural eotaxin 1504:1518 natural eotaxin 1504:1518 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 3 64 from mass 639:642 arg1 electrophoresis 680:694 SDS/polyacrylamide gel electrophoresis 657:694 SDS/polyacrylamide gel electrophoresis 657:694 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 5 65 theme N-terminally 832:843 arg1 forms 855:859 50% N-terminally truncated forms 828:859 50% N-terminally truncated forms missing two or three amino acids 828:892 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 8 66 theme eosinophil 1174:1183 arg1 chemotaxis 1185:1194 eosinophil chemotaxis 1174:1194 eosinophil chemotaxis 1174:1194 Natural eotaxin stimulated eosinophil chemotaxis with identical potency and efficacy as recombinant human eotaxin. 9578468 7 67 theme principal 1106:1114 arg1 masses 1116:1121 principal masses 1106:1121 principal masses of 9033 Da and 9317 Da 1106:1144 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 10 68 theme monocyte 1641:1648 arg1 protein 1662:1668 monocyte chemotactic protein 3 1641:1670 monocyte chemotactic protein 3 (MCP-3) 1641:1678 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 10 68 theme monocyte 1641:1648 arg1 MCP-3 1673:1677 MCP-3 1673:1677 MCP-3 1673:1677 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 10 69 theme eosinophils 1487:1497 arg1 Treatment 1474:1482 Treatment 1474:1482 Treatment of eosinophils with natural eotaxin 1474:1518 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 3 70 theme activity 568:575 arg1 Purification 547:558 Purification 547:558 Purification of this activity 547:575 Purification of this activity revealed a heparin-binding protein with an apparent molecular mass of 13 kDa in SDS/polyacrylamide gel electrophoresis. 9578468 2 71 theme eosinophil-specific 505:523 arg1 activity 537:544 eosinophil-specific chemotactic activity 505:544 eosinophil-specific chemotactic activity 505:544 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 6 72 gly O-glycosylated 963:976 arg1 variants 978:985 at least two sialylated O-glycosylated variants 939:985 at least two sialylated O-glycosylated variants 939:985 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 1 73 theme potential 328:336 arg1 source 347:352 a potential cellular source 326:352 a potential cellular source of eosinophil-selective attractants 326:388 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 7 74 theme eotaxin 1061:1067 arg1 preparation 1069:1079 the natural eotaxin preparation 1049:1079 the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da 1049:1144 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 2 75 theme stimulation 492:502 arg1 day 483:485 day 3 483:487 day 3 of stimulation 483:502 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 5 76 contain contain 820:826 arg1 preparations 807:818 Natural eotaxin preparations 791:818 Natural eotaxin preparations 791:818 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 5 76 contain contain 820:826 arg2 forms 855:859 50% N-terminally truncated forms 828:859 50% N-terminally truncated forms missing two or three amino acids 828:892 Natural eotaxin preparations contain 50% N-terminally truncated forms missing two or three amino acids. 9578468 10 77 theme natural 1504:1510 arg1 eotaxin 1512:1518 natural eotaxin 1504:1518 natural eotaxin 1504:1518 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 0 78 theme O-glycosylated 42:55 arg1 forms 57:61 biologically active O-glycosylated forms 22:61 biologically active O-glycosylated forms of human eotaxin 22:78 Delayed production of biologically active O-glycosylated forms of human eotaxin by tumor-necrosis-factor-alpha-stimulated dermal fibroblasts. 9578468 8 79 theme recombinant 1235:1245 arg1 eotaxin 1253:1259 recombinant human eotaxin 1235:1259 recombinant human eotaxin 1235:1259 Natural eotaxin stimulated eosinophil chemotaxis with identical potency and efficacy as recombinant human eotaxin. 9578468 6 80 theme O-glycosylated 963:976 arg1 variants 978:985 at least two sialylated O-glycosylated variants 939:985 at least two sialylated O-glycosylated variants 939:985 It is O-glycosylated at Thr71, resulting in at least two sialylated O-glycosylated variants. 9578468 1 81 theme eosinophil-selective 357:376 arg1 attractants 378:388 eosinophil-selective attractants 357:388 eosinophil-selective attractants 357:388 Since a number of inflammatory skin diseases are characterized by selective eosinophil infiltration preferentially in the dermis, we speculated that dermal fibroblasts might represent a potential cellular source of eosinophil-selective attractants. 9578468 10 82 theme normal 1585:1590 arg1 RANTES 1629:1634 RANTES 1629:1634 RANTES 1629:1634 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 10 82 theme normal 1585:1590 arg1 T-lymphocyte 1592:1603 an normal T-lymphocyte 1582:1603 an normal T-lymphocyte expressed and secreted (RANTES) 1582:1635 Treatment of eosinophils with natural eotaxin desensitizes chemotactic responses towards eotaxin, regulated an normal T-lymphocyte expressed and secreted (RANTES) and monocyte chemotactic protein 3 (MCP-3), whereas RANTES and MCP-3 were unable to desensitize natural eotaxin-dependent responses. 9578468 2 83 theme Cultivated 391:400 arg1 fibroblasts 409:419 Cultivated dermal fibroblasts 391:419 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha 391:460 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 7 84 theme spectrometric 1017:1029 arg1 analyses 1031:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 9578468 4 85 theme amino 729:733 arg1 analyses 749:756 subsequent amino acid sequence analyses 718:756 subsequent amino acid sequence analyses 718:756 Peptide mapping with subsequent amino acid sequence analyses revealed it to be human eotaxin. 9578468 2 86 theme necrosis 440:447 arg1 alpha 456:460 tumor necrosis factor alpha 434:460 tumor necrosis factor alpha 434:460 Cultivated dermal fibroblasts treated with tumor necrosis factor alpha secreted, not before day 3 of stimulation, eosinophil-specific chemotactic activity. 9578468 7 87 theme ionization 1001:1010 arg1 analyses 1031:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses 988:1038 Electrospray ionization mass spectrometric analyses revealed the natural eotaxin preparation to be heterogeneous with principal masses of 9033 Da and 9317 Da. 19879838 1 0 theme HIV-1 153:157 arg1 release 142:148 the release 138:148 the release of HIV-1 and other enveloped viral particles 138:193 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 5 1 theme native 680:685 arg1 tetherin 687:694 native tetherin 680:694 native tetherin 680:694 These findings allowed the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity. 19879838 4 2 theme primary 526:532 arg1 sequence 534:541 primary sequence 526:541 tetherin configuration rather than primary sequence 491:541 We demonstrate, through mutational analyses and domain replacement experiments, that tetherin configuration rather than primary sequence is critical for antiviral activity. 19879838 2 3 theme virion 294:299 arg1 retention 301:309 virion retention 294:309 virion retention 294:309 The underlying mechanism by which tetherin functions and whether it directly or indirectly causes virion retention are unknown. 19879838 7 4 theme plasma 1136:1141 arg1 membrane 1143:1150 the plasma membrane 1132:1150 the plasma membrane 1132:1150 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19879838 5 5 theme artificial 629:638 arg1 protein 640:646 a completely artificial protein 616:646 a completely artificial protein 616:646 These findings allowed the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity. 19879838 5 6 theme protein 640:646 arg1 design 606:611 the design 602:611 the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity 602:745 These findings allowed the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity. 19879838 1 7 theme other 163:167 arg1 particles 185:193 other enveloped viral particles 163:193 other enveloped viral particles 163:193 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 6 8 theme parallel 820:827 arg1 tetherin 769:776 tetherin 769:776 tetherin 769:776 We further show that tetherin is incorporated into HIV-1 particles as a parallel homodimer using either of its two membrane anchors. 19879838 6 8 theme parallel 820:827 arg1 homodimer 829:837 a parallel homodimer 818:837 a parallel homodimer using either of its two membrane anchors 818:878 We further show that tetherin is incorporated into HIV-1 particles as a parallel homodimer using either of its two membrane anchors. 19879838 7 9 dep necessary 1045:1053 arg1 sufficient 1067:1076 sufficient 1067:1076 sufficient 1067:1076 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19879838 1 10 theme enveloped 169:177 arg1 particles 185:193 other enveloped viral particles 163:193 other enveloped viral particles 163:193 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 4 11 theme tetherin 491:498 arg1 configuration 500:512 tetherin configuration 491:512 tetherin configuration rather than primary sequence 491:541 We demonstrate, through mutational analyses and domain replacement experiments, that tetherin configuration rather than primary sequence is critical for antiviral activity. 19879838 5 12 theme sequence 657:664 arg1 homology 666:673 sequence homology 657:673 sequence homology with native tetherin 657:694 These findings allowed the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity. 19879838 7 13 theme envelopes 986:994 arg1 infiltration 963:974 infiltration 963:974 infiltration of virion envelopes by one or both of tetherin's membrane anchors 963:1040 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19879838 1 14 theme viral 179:183 arg1 particles 185:193 other enveloped viral particles 163:193 other enveloped viral particles 163:193 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 3 15 theme antiviral 386:394 arg1 activity 396:403 its antiviral activity 382:403 its antiviral activity 382:403 Here, we elucidate the mechanism by which tetherin exerts its antiviral activity. 19879838 1 16 theme particles 185:193 arg1 release 142:148 the release 138:148 the release of HIV-1 and other enveloped viral particles 138:193 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 5 17 theme antiviral 728:736 arg1 activity 738:745 its antiviral activity 724:745 its antiviral activity 724:745 These findings allowed the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity. 19879838 1 18 theme interferon-induced 87:104 arg1 Tetherin 72:79 Tetherin 72:79 Tetherin 72:79 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 1 18 theme interferon-induced 87:104 arg1 protein 106:112 an interferon-induced protein 84:112 an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles 84:193 Tetherin is an interferon-induced protein whose expression blocks the release of HIV-1 and other enveloped viral particles. 19879838 0 19 theme HIV-1 18:22 arg1 release 24:30 HIV-1 release 18:30 HIV-1 release 18:30 Tetherin inhibits HIV-1 release by directly tethering virions to cells. 19879838 7 20 theme virus 1099:1103 arg1 particles 1105:1113 enveloped virus particles 1089:1113 enveloped virus particles that bud 1089:1122 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19879838 4 21 theme domain 454:459 arg1 experiments 473:483 domain replacement experiments 454:483 domain replacement experiments 454:483 We demonstrate, through mutational analyses and domain replacement experiments, that tetherin configuration rather than primary sequence is critical for antiviral activity. 19879838 7 22 theme virion 979:984 arg1 envelopes 986:994 virion envelopes 979:994 virion envelopes 979:994 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19879838 6 23 theme HIV-1 799:803 arg1 particles 805:813 HIV-1 particles 799:813 HIV-1 particles 799:813 We further show that tetherin is incorporated into HIV-1 particles as a parallel homodimer using either of its two membrane anchors. 19879838 4 24 theme replacement 461:471 arg1 experiments 473:483 domain replacement experiments 454:483 domain replacement experiments 454:483 We demonstrate, through mutational analyses and domain replacement experiments, that tetherin configuration rather than primary sequence is critical for antiviral activity. 19879838 4 25 theme antiviral 559:567 arg1 activity 569:576 antiviral activity 559:576 antiviral activity 559:576 We demonstrate, through mutational analyses and domain replacement experiments, that tetherin configuration rather than primary sequence is critical for antiviral activity. 19879838 7 26 theme enveloped 1089:1097 arg1 particles 1105:1113 enveloped virus particles 1089:1113 enveloped virus particles that bud 1089:1122 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19879838 6 27 theme membrane 863:870 arg1 anchors 872:878 its two membrane anchors 855:878 its two membrane anchors 855:878 We further show that tetherin is incorporated into HIV-1 particles as a parallel homodimer using either of its two membrane anchors. 19879838 5 28 with homology 666:673 arg1 tetherin 687:694 native tetherin 680:694 native tetherin 680:694 These findings allowed the design of a completely artificial protein, lacking sequence homology with native tetherin, that nevertheless mimicked its antiviral activity. 19879838 2 29 theme underlying 200:209 arg1 unknown 315:321 unknown 315:321 unknown 315:321 The underlying mechanism by which tetherin functions and whether it directly or indirectly causes virion retention are unknown. 19879838 2 29 theme underlying 200:209 arg1 mechanism 211:219 The underlying mechanism 196:219 The underlying mechanism by which tetherin functions and whether it directly or indirectly causes virion retention 196:309 The underlying mechanism by which tetherin functions and whether it directly or indirectly causes virion retention are unknown. 19879838 4 30 theme mutational 430:439 arg1 analyses 441:448 mutational analyses 430:448 mutational analyses 430:448 We demonstrate, through mutational analyses and domain replacement experiments, that tetherin configuration rather than primary sequence is critical for antiviral activity. 19879838 7 31 theme membrane 1025:1032 arg1 anchors 1034:1040 tetherin's membrane anchors 1014:1040 tetherin's membrane anchors 1014:1040 These results indicate that tetherin functions autonomously and directly and that infiltration of virion envelopes by one or both of tetherin's membrane anchors is necessary, and likely sufficient, to tether enveloped virus particles that bud through the plasma membrane. 19880749 3 0 dep residues 408:415 arg1 287-685 417:423 287-685 417:423 287-685 417:423 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 7 1 theme D 1032:1032 arg1 domains 1047:1053 the D, C(A), and S domains 1028:1053 the D, C(A), and S domains traversing the W-shaped molecule 1028:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 1 2 theme Willebrand 166:175 arg1 VWF 185:187 VWF 185:187 VWF 185:187 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 1 2 theme Willebrand 166:175 arg1 factor 177:182 plasma von Willebrand factor 155:182 plasma von Willebrand factor (VWF) 155:188 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 5 3 theme 10-stranded 737:747 arg1 fold 763:766 a 10-stranded beta-sandwich fold 735:766 a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain 735:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 0 4 theme discontinuous 75:87 arg1 exosites 89:96 multiple discontinuous exosites 66:96 multiple discontinuous exosites for von Willebrand factor 66:122 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 4 5 theme disintegrin-like 562:577 arg1 domain 638:643 the cysteine-rich domain 620:643 the cysteine-rich domain (designated the C(A) domain) 620:672 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 4 5 theme disintegrin-like 562:577 arg1 domain 583:588 the disintegrin-like (D) domain 558:588 the disintegrin-like (D) domain 558:588 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 4 5 theme disintegrin-like 562:577 arg1 D 580:580 D 580:580 D 580:580 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 7 6 theme W-shaped 1070:1077 arg1 molecule 1079:1086 the W-shaped molecule 1066:1086 the W-shaped molecule 1066:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 8 7 theme enzyme-substrate 1198:1213 arg1 interactions 1215:1226 the multiple enzyme-substrate interactions 1185:1226 the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system 1185:1264 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 0 8 theme von 102:104 arg1 factor 117:122 von Willebrand factor 102:122 von Willebrand factor 102:122 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 7 9 theme VWF-binding 957:967 arg1 exosites 969:976 3 VWF-binding exosites 955:976 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule 955:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 2 10 theme thrombocytopenic 323:338 arg1 purpura 340:346 life-threatening thrombotic thrombocytopenic purpura 295:346 life-threatening thrombotic thrombocytopenic purpura 295:346 Severe deficiencies in ADAMTS13 can cause life-threatening thrombotic thrombocytopenic purpura. 19880749 4 11 theme domain 638:643 arg1 domain 638:643 the cysteine-rich domain 620:643 the cysteine-rich domain (designated the C(A) domain) 620:672 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 4 11 theme domain 638:643 arg1 domain 583:588 the disintegrin-like (D) domain 558:588 the disintegrin-like (D) domain 558:588 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 4 11 theme domain 638:643 arg1 D 580:580 D 580:580 D 580:580 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 4 11 theme domain 638:643 arg1 portion 609:615 the N-terminal portion 594:615 the N-terminal portion of the cysteine-rich domain (designated the C(A) domain) 594:672 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 5 12 theme spacer 679:684 arg1 domain 690:695 The spacer (S) domain 675:695 The spacer (S) domain 675:695 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 2 13 theme thrombotic 312:321 arg1 purpura 340:346 life-threatening thrombotic thrombocytopenic purpura 295:346 life-threatening thrombotic thrombocytopenic purpura 295:346 Severe deficiencies in ADAMTS13 can cause life-threatening thrombotic thrombocytopenic purpura. 19880749 4 14 theme cysteine-rich 624:636 arg1 domain 638:643 the cysteine-rich domain 620:643 the cysteine-rich domain (designated the C(A) domain) 620:672 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 8 15 theme ADAMTS 1133:1138 arg1 proteins 1147:1154 ADAMTS family proteins 1133:1154 ADAMTS family proteins 1133:1154 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 7 16 theme domains 1047:1053 arg1 surfaces 1016:1023 the linearly aligned discontinuous surfaces 981:1023 the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule 981:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 2 17 theme life-threatening 295:310 arg1 purpura 340:346 life-threatening thrombotic thrombocytopenic purpura 295:346 life-threatening thrombotic thrombocytopenic purpura 295:346 Severe deficiencies in ADAMTS13 can cause life-threatening thrombotic thrombocytopenic purpura. 19880749 5 18 theme interaction 786:796 arg1 sites 798:802 multiple interaction sites 777:802 multiple interaction sites 777:802 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 0 19 theme Willebrand 106:115 arg1 factor 117:122 von Willebrand factor 102:122 von Willebrand factor 102:122 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 4 20 theme folding 529:535 arg1 similarities 537:548 folding similarities 529:548 folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain) 529:672 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 3 21 theme human 449:453 arg1 fragment 464:471 an exosite-containing human ADAMTS13 fragment 427:471 an exosite-containing human ADAMTS13 fragment 427:471 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 3 21 theme human 449:453 arg1 ADAMTS13-DTCS 393:405 ADAMTS13-DTCS 393:405 ADAMTS13-DTCS (residues 287-685) 393:424 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 5 22 theme C 813:813 arg1 domain 818:823 the C(A) domain 809:823 the C(A) domain 809:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 3 23 theme 2.6-A 477:481 arg1 resolution 493:502 2.6-A and 2.8-A resolution 477:502 resolution 493:502 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 8 24 theme MDTCS 1099:1103 arg1 domains 1105:1111 the MDTCS domains 1095:1111 the MDTCS domains 1095:1111 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 5 25 theme beta-sandwich 749:761 arg1 fold 763:766 a 10-stranded beta-sandwich fold 735:766 a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain 735:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 8 26 theme proteinases 1342:1352 arg1 class 1333:1337 this class 1328:1337 this class of proteinases 1328:1352 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 7 27 theme C 1035:1035 arg1 domains 1047:1053 the D, C(A), and S domains 1028:1053 the D, C(A), and S domains traversing the W-shaped molecule 1028:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 0 28 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of the noncatalytic domains of ADAMTS13 0:57 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 7 29 theme S 1045:1045 arg1 domains 1047:1053 the D, C(A), and S domains 1028:1053 the D, C(A), and S domains traversing the W-shaped molecule 1028:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 8 30 theme interactions 1215:1226 arg1 framework 1172:1180 the structural framework 1157:1180 the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system 1157:1264 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 8 31 theme substrate 1298:1306 arg1 mode 1320:1323 a common substrate recognition mode 1289:1323 a common substrate recognition mode in this class of proteinases 1289:1352 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 2 32 from deficiencies 260:271 arg1 ADAMTS13 276:283 ADAMTS13 276:283 ADAMTS13 276:283 Severe deficiencies in ADAMTS13 can cause life-threatening thrombotic thrombocytopenic purpura. 19880749 4 33 theme A 663:663 arg1 domain 666:671 the C(A) domain 657:671 the C(A) domain 657:671 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 5 34 theme A 815:815 arg1 domain 818:823 the C(A) domain 809:823 the C(A) domain 809:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 5 35 theme functional 714:723 arg1 unit 725:728 a globular functional unit 703:728 a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain 703:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 8 36 theme common 1291:1296 arg1 mode 1320:1323 a common substrate recognition mode 1289:1323 a common substrate recognition mode in this class of proteinases 1289:1352 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 0 37 theme domains 39:45 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of the noncatalytic domains of ADAMTS13 0:57 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 4 38 theme N-terminal 598:607 arg1 portion 609:615 the N-terminal portion 594:615 the N-terminal portion of the cysteine-rich domain (designated the C(A) domain) 594:672 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 5 39 theme globular 705:712 arg1 unit 725:728 a globular functional unit 703:728 a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain 703:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 6 40 theme structure-based 842:856 arg1 mutants 858:864 25 structure-based mutants 839:864 25 structure-based mutants of ADAMTS13-MDTCS (residues 75-685) 839:900 We expressed 25 structure-based mutants of ADAMTS13-MDTCS (residues 75-685) and measured their enzymatic activity. 19880749 2 41 theme Severe 253:258 arg1 deficiencies 260:271 Severe deficiencies 253:271 Severe deficiencies in ADAMTS13 253:283 Severe deficiencies in ADAMTS13 can cause life-threatening thrombotic thrombocytopenic purpura. 19880749 8 42 from mode 1320:1323 arg1 class 1333:1337 this class 1328:1337 this class of proteinases 1328:1352 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 1 43 theme VWF-mediated 211:222 arg1 formation 242:250 VWF-mediated platelet thrombus formation 211:250 VWF-mediated platelet thrombus formation 211:250 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 0 44 theme noncatalytic 26:37 arg1 domains 39:45 the noncatalytic domains 22:45 the noncatalytic domains of ADAMTS13 22:57 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 0 44 theme noncatalytic 26:37 arg1 ADAMTS13 50:57 ADAMTS13 50:57 ADAMTS13 50:57 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 3 45 theme ADAMTS13 455:462 arg1 fragment 464:471 an exosite-containing human ADAMTS13 fragment 427:471 an exosite-containing human ADAMTS13 fragment 427:471 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 3 45 theme ADAMTS13 455:462 arg1 ADAMTS13-DTCS 393:405 ADAMTS13-DTCS 393:405 ADAMTS13-DTCS (residues 287-685) 393:424 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 6 46 dep residues 885:892 arg1 75-685 894:899 75-685 894:899 75-685 894:899 We expressed 25 structure-based mutants of ADAMTS13-MDTCS (residues 75-685) and measured their enzymatic activity. 19880749 1 47 theme platelet 224:231 arg1 formation 242:250 VWF-mediated platelet thrombus formation 211:250 VWF-mediated platelet thrombus formation 211:250 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 0 48 theme ADAMTS13 50:57 arg1 domains 39:45 the noncatalytic domains 22:45 the noncatalytic domains of ADAMTS13 22:57 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 0 48 theme ADAMTS13 50:57 arg1 ADAMTS13 50:57 ADAMTS13 50:57 ADAMTS13 50:57 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 3 49 theme ADAMTS13-DTCS 393:405 arg1 structures 379:388 2 crystal structures 369:388 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, 369:472 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 8 50 theme structural 1161:1170 arg1 framework 1172:1180 the structural framework 1157:1180 the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system 1157:1264 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 5 51 with unit 725:728 arg1 fold 763:766 a 10-stranded beta-sandwich fold 735:766 a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain 735:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 6 52 theme ADAMTS13-MDTCS 869:882 arg1 mutants 858:864 25 structure-based mutants 839:864 25 structure-based mutants of ADAMTS13-MDTCS (residues 75-685) 839:900 We expressed 25 structure-based mutants of ADAMTS13-MDTCS (residues 75-685) and measured their enzymatic activity. 19880749 3 53 theme exosite-containing 430:447 arg1 fragment 464:471 an exosite-containing human ADAMTS13 fragment 427:471 an exosite-containing human ADAMTS13 fragment 427:471 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 3 53 theme exosite-containing 430:447 arg1 ADAMTS13-DTCS 393:405 ADAMTS13-DTCS 393:405 ADAMTS13-DTCS (residues 287-685) 393:424 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 6 54 theme enzymatic 921:929 arg1 activity 931:938 their enzymatic activity 915:938 their enzymatic activity 915:938 We expressed 25 structure-based mutants of ADAMTS13-MDTCS (residues 75-685) and measured their enzymatic activity. 19880749 5 55 theme S 687:687 arg1 domain 690:695 The spacer (S) domain 675:695 The spacer (S) domain 675:695 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 7 56 theme aligned 994:1000 arg1 surfaces 1016:1023 the linearly aligned discontinuous surfaces 981:1023 the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule 981:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 7 57 theme discontinuous 1002:1014 arg1 surfaces 1016:1023 the linearly aligned discontinuous surfaces 981:1023 the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule 981:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 7 58 from exosites 969:976 arg1 surfaces 1016:1023 the linearly aligned discontinuous surfaces 981:1023 the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule 981:1086 We identified 3 VWF-binding exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S domains traversing the W-shaped molecule. 19880749 1 59 theme thrombus 233:240 arg1 formation 242:250 VWF-mediated platelet thrombus formation 211:250 VWF-mediated platelet thrombus formation 211:250 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 5 60 theme multiple 777:784 arg1 sites 798:802 multiple interaction sites 777:802 multiple interaction sites 777:802 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 8 61 theme family 1140:1145 arg1 proteins 1147:1154 ADAMTS family proteins 1133:1154 ADAMTS family proteins 1133:1154 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 8 62 theme recognition 1308:1318 arg1 mode 1320:1323 a common substrate recognition mode 1289:1323 a common substrate recognition mode in this class of proteinases 1289:1352 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 0 63 theme multiple 66:73 arg1 exosites 89:96 multiple discontinuous exosites 66:96 multiple discontinuous exosites for von Willebrand factor 66:122 Crystal structures of the noncatalytic domains of ADAMTS13 reveal multiple discontinuous exosites for von Willebrand factor. 19880749 3 64 theme 2.8-A 487:491 arg1 resolution 493:502 2.6-A and 2.8-A resolution 477:502 resolution 493:502 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 8 65 theme ADAMTS13-VWF 1246:1257 arg1 system 1259:1264 the ADAMTS13-VWF system 1242:1264 the ADAMTS13-VWF system 1242:1264 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 1 66 theme plasma 155:160 arg1 VWF 185:187 VWF 185:187 VWF 185:187 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 1 66 theme plasma 155:160 arg1 factor 177:182 plasma von Willebrand factor 155:182 plasma von Willebrand factor (VWF) 155:188 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 8 67 theme multiple 1189:1196 arg1 interactions 1215:1226 the multiple enzyme-substrate interactions 1185:1226 the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system 1185:1264 Since the MDTCS domains are conserved among ADAMTS family proteins, the structural framework of the multiple enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the basis for a common substrate recognition mode in this class of proteinases. 19880749 4 68 theme C 661:661 arg1 domain 666:671 the C(A) domain 657:671 the C(A) domain 657:671 The structures revealed folding similarities between the disintegrin-like (D) domain and the N-terminal portion of the cysteine-rich domain (designated the C(A) domain). 19880749 3 69 theme crystal 371:377 arg1 structures 379:388 2 crystal structures 369:388 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, 369:472 Here, we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. 19880749 5 70 contain has 773:775 arg1 fold 763:766 a 10-stranded beta-sandwich fold 735:766 a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain 735:823 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 5 70 contain has 773:775 arg2 sites 798:802 multiple interaction sites 777:802 multiple interaction sites 777:802 The spacer (S) domain forms a globular functional unit with a 10-stranded beta-sandwich fold that has multiple interaction sites with the C(A) domain. 19880749 1 71 theme von 162:164 arg1 VWF 185:187 VWF 185:187 VWF 185:187 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 19880749 1 71 theme von 162:164 arg1 factor 177:182 plasma von Willebrand factor 155:182 plasma von Willebrand factor (VWF) 155:188 ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby controls VWF-mediated platelet thrombus formation. 16905106 7 0 with interactions 1021:1032 arg1 receptors 1039:1047 receptors 1039:1047 receptors 1039:1047 These structures demonstrate the structural plasticity of TNFSF members and their interactions with receptors. 16905106 3 1 theme murine 331:336 arg1 OX40L 338:342 murine OX40L 331:342 murine OX40L 331:342 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 4 2 theme small 462:466 arg1 OX40L 440:444 OX40L 440:444 OX40L 440:444 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 4 2 theme small 462:466 arg1 member 468:473 an unusually small member 449:473 an unusually small member of the tumor necrosis factor superfamily (TNFSF) 449:522 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 3 3 theme OX40L 338:342 arg1 structure 318:326 the crystal structure 306:326 the crystal structure of murine OX40L and of the human OX40-OX40L complex 306:378 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 2 4 theme interaction 140:150 arg1 Blockade 113:120 Blockade 113:120 Blockade of the OX40L-OX40 interaction 113:150 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 5 5 theme OX40L 548:552 arg1 protomers 554:562 the OX40L protomers 544:562 the OX40L protomers forming the functional trimer 544:592 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 6 6 theme binding 877:883 arg1 energy 885:890 binding energy 877:890 binding energy 877:890 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 2 7 theme OX40L-OX40 129:138 arg1 interaction 140:150 the OX40L-OX40 interaction 125:150 the OX40L-OX40 interaction 125:150 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 7 8 theme TNFSF 997:1001 arg1 members 1003:1009 TNFSF members 997:1009 TNFSF members 997:1009 These structures demonstrate the structural plasticity of TNFSF members and their interactions with receptors. 16905106 5 9 theme protomers 554:562 arg1 atypical 597:604 atypical 597:604 atypical 597:604 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 5 9 theme protomers 554:562 arg1 arrangement 529:539 The arrangement 525:539 The arrangement of the OX40L protomers forming the functional trimer 525:592 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 2 10 contain has 152:154 arg1 Blockade 113:120 Blockade 113:120 Blockade of the OX40L-OX40 interaction 113:150 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 2 10 contain has 152:154 arg2 effects 169:175 ameliorative effects 156:175 ameliorative effects 156:175 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 3 11 dep understand 236:245 arg1 to 226:227 to 226:227 to 226:227 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 6 12 theme OX40L 804:808 arg1 OX40L 804:808 OX40L 804:808 OX40L 804:808 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 6 12 theme OX40L 804:808 arg1 residues 792:799 the interfacial residues 776:799 the interfacial residues of OX40L 776:808 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 7 13 theme members 1003:1009 arg1 plasticity 983:992 the structural plasticity 968:992 the structural plasticity of TNFSF members 968:1009 These structures demonstrate the structural plasticity of TNFSF members and their interactions with receptors. 16905106 7 13 theme members 1003:1009 arg1 interactions 1021:1032 their interactions 1015:1032 their interactions with receptors 1015:1047 These structures demonstrate the structural plasticity of TNFSF members and their interactions with receptors. 16905106 6 14 theme hot 849:851 arg1 " 857:857 a single "hot spot" 839:857 a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas 839:936 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 0 15 theme crystal 4:10 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of the costimulatory OX40-OX40L complex 0:60 The crystal structure of the costimulatory OX40-OX40L complex. 16905106 5 16 theme functional 576:585 arg1 trimer 587:592 the functional trimer 572:592 the functional trimer 572:592 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 1 17 theme T 73:73 arg1 costimulator 80:91 a T cell costimulator 71:91 a T cell costimulator activated by OX40L 71:110 OX40 is a T cell costimulator activated by OX40L. 16905106 1 17 theme T 73:73 arg1 OX40 63:66 OX40 63:66 OX40 63:66 OX40 is a T cell costimulator activated by OX40L. 16905106 3 18 theme human 355:359 arg1 complex 372:378 the human OX40-OX40L complex 351:378 the human OX40-OX40L complex 351:378 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 4 19 theme superfamily 504:514 arg1 OX40L 440:444 OX40L 440:444 OX40L 440:444 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 4 19 theme superfamily 504:514 arg1 member 468:473 an unusually small member 449:473 an unusually small member of the tumor necrosis factor superfamily (TNFSF) 449:522 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 1 20 theme cell 75:78 arg1 costimulator 80:91 a T cell costimulator 71:91 a T cell costimulator activated by OX40L 71:110 OX40 is a T cell costimulator activated by OX40L. 16905106 1 20 theme cell 75:78 arg1 OX40 63:66 OX40 63:66 OX40 63:66 OX40 is a T cell costimulator activated by OX40L. 16905106 0 21 theme OX40-OX40L 43:52 arg1 complex 54:60 the costimulatory OX40-OX40L complex 25:60 the costimulatory OX40-OX40L complex 25:60 The crystal structure of the costimulatory OX40-OX40L complex. 16905106 4 22 theme factor 497:502 arg1 superfamily 504:514 tumor necrosis factor superfamily 482:514 the tumor necrosis factor superfamily (TNFSF) 478:522 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 4 22 theme factor 497:502 arg1 TNFSF 517:521 TNFSF 517:521 TNFSF 517:521 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 5 23 theme open 736:739 arg1 assembly 741:748 an open assembly 733:748 an open assembly 733:748 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 7 24 theme structural 972:981 arg1 plasticity 983:992 the structural plasticity 968:992 the structural plasticity of TNFSF members 968:1009 These structures demonstrate the structural plasticity of TNFSF members and their interactions with receptors. 16905106 6 25 theme residues 792:799 arg1 changes 765:771 Site-directed changes 751:771 Site-directed changes of the interfacial residues of OX40L 751:808 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 0 26 theme costimulatory 29:41 arg1 complex 54:60 the costimulatory OX40-OX40L complex 25:60 the costimulatory OX40-OX40L complex 25:60 The crystal structure of the costimulatory OX40-OX40L complex. 16905106 6 27 theme interfacial 780:790 arg1 OX40L 804:808 OX40L 804:808 OX40L 804:808 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 6 27 theme interfacial 780:790 arg1 residues 792:799 the interfacial residues 776:799 the interfacial residues of OX40L 776:808 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 5 28 theme trimer 707:712 arg1 axis 714:717 the trimer axis 703:717 the trimer axis 703:717 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 2 29 theme ameliorative 156:167 arg1 effects 169:175 ameliorative effects 156:175 ameliorative effects 156:175 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 6 30 theme distinct 923:930 arg1 areas 932:936 at least two distinct areas 910:936 at least two distinct areas 910:936 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 3 31 theme OX40-OX40L 361:370 arg1 complex 372:378 the human OX40-OX40L complex 351:378 the human OX40-OX40L complex 351:378 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 5 32 theme degrees 653:659 arg1 rotation 661:668 a 15 degrees rotation 648:668 a 15 degrees rotation of each protomer 648:685 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 3 33 theme crystal 310:316 arg1 structure 318:326 the crystal structure 306:326 the crystal structure of murine OX40L and of the human OX40-OX40L complex 306:378 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 2 34 theme pathologies 204:214 arg1 models 187:192 animal models 180:192 animal models of T cell pathologies 180:214 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 7 35 with plasticity 983:992 arg1 receptors 1039:1047 receptors 1039:1047 receptors 1039:1047 These structures demonstrate the structural plasticity of TNFSF members and their interactions with receptors. 16905106 3 36 theme complex 372:378 arg1 structure 318:326 the crystal structure 306:326 the crystal structure of murine OX40L and of the human OX40-OX40L complex 306:378 In order to better understand the interaction between OX40 and OX40L, we have determined the crystal structure of murine OX40L and of the human OX40-OX40L complex at 1.45 and 2.4 A, respectively. 16905106 4 37 theme necrosis 488:495 arg1 superfamily 504:514 tumor necrosis factor superfamily 482:514 the tumor necrosis factor superfamily (TNFSF) 478:522 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 4 37 theme necrosis 488:495 arg1 TNFSF 517:521 TNFSF 517:521 TNFSF 517:521 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 2 38 theme T 197:197 arg1 pathologies 204:214 T cell pathologies 197:214 T cell pathologies 197:214 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 2 39 theme cell 199:202 arg1 pathologies 204:214 T cell pathologies 197:214 T cell pathologies 197:214 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 4 40 theme tumor 482:486 arg1 superfamily 504:514 tumor necrosis factor superfamily 482:514 the tumor necrosis factor superfamily (TNFSF) 478:522 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 4 40 theme tumor 482:486 arg1 TNFSF 517:521 TNFSF 517:521 TNFSF 517:521 These structures show that OX40L is an unusually small member of the tumor necrosis factor superfamily (TNFSF). 16905106 6 41 theme Site-directed 751:763 arg1 changes 765:771 Site-directed changes 751:771 Site-directed changes of the interfacial residues of OX40L 751:808 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 5 42 theme other 631:635 arg1 members 637:643 other members 631:643 other members 631:643 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 5 43 theme protomer 678:685 arg1 rotation 661:668 a 15 degrees rotation 648:668 a 15 degrees rotation of each protomer 648:685 The arrangement of the OX40L protomers forming the functional trimer is atypical and differs from that of other members by a 15 degrees rotation of each protomer with respect to the trimer axis, resulting in an open assembly. 16905106 2 44 theme animal 180:185 arg1 models 187:192 animal models 180:192 animal models of T cell pathologies 180:214 Blockade of the OX40L-OX40 interaction has ameliorative effects in animal models of T cell pathologies. 16905106 0 45 theme complex 54:60 arg1 structure 12:20 The crystal structure 0:20 The crystal structure of the costimulatory OX40-OX40L complex 0:60 The crystal structure of the costimulatory OX40-OX40L complex. 16905106 6 46 theme single 841:846 arg1 " 857:857 a single "hot spot" 839:857 a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas 839:936 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16905106 6 47 theme spot 853:856 arg1 " 857:857 a single "hot spot" 839:857 a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas 839:936 Site-directed changes of the interfacial residues of OX40L suggest this interface lacks a single "hot spot" and that instead, binding energy is dispersed over at least two distinct areas. 16765900 1 0 theme populations 303:313 arg1 maintenance 274:284 maintenance 274:284 maintenance 274:284 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 0 theme populations 303:313 arg1 development 258:268 development 258:268 development 258:268 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 2 1 theme RET 554:556 arg1 receptor 558:565 the shared RET receptor 543:565 the shared RET receptor 543:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 5 2 theme therapeutic 1113:1123 arg1 applications 1125:1136 therapeutic applications 1113:1136 therapeutic applications 1113:1136 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 3 3 theme specificity-determining 682:704 arg1 points 713:718 both conserved and specificity-determining anchor points 663:718 both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 663:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 1 4 theme ligands 224:230 arg1 Artemin 132:138 Artemin 132:138 Artemin (ARTN) 132:145 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 4 theme ligands 224:230 arg1 member 152:157 a member 150:157 a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system 150:345 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 5 dep development 258:268 arg1 the 254:256 the 254:256 the 254:256 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 2 6 theme shared 547:552 arg1 receptor 558:565 the shared RET receptor 543:565 the shared RET receptor 543:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 1 7 from maintenance 274:284 arg1 system 340:345 the mammalian nervous system 318:345 the mammalian nervous system 318:345 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 5 8 from exploitation 1096:1107 arg1 neurobiology 1067:1078 neurobiology 1067:1078 neurobiology 1067:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 0 9 theme cell 92:95 arg1 factors 123:129 glial cell line-derived neurotrophic factors 86:129 glial cell line-derived neurotrophic factors 86:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 5 10 theme roles 1043:1047 arg1 dissection 997:1006 the functional dissection 982:1006 the functional dissection of the specific versus pleiotropic roles of this system in neurobiology 982:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 5 10 theme roles 1043:1047 arg1 exploitation 1096:1107 its exploitation 1092:1107 its exploitation for therapeutic applications 1092:1136 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 0 11 theme glial 86:90 arg1 factors 123:129 glial cell line-derived neurotrophic factors 86:129 glial cell line-derived neurotrophic factors 86:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 1 12 theme mammalian 322:330 arg1 system 340:345 the mammalian nervous system 318:345 the mammalian nervous system 318:345 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 13 link line-derived 177:188 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 13 link line-derived 177:188 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 0 14 theme neurotrophic 110:121 arg1 factors 123:129 glial cell line-derived neurotrophic factors 86:129 glial cell line-derived neurotrophic factors 86:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 1 15 theme nervous 332:338 arg1 system 340:345 the mammalian nervous system 318:345 the mammalian nervous system 318:345 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 2 16 theme crystal 374:380 arg1 structure 382:390 the 1.92 A crystal structure 363:390 the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor 363:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 0 17 theme line-derived 97:108 arg1 factors 123:129 glial cell line-derived neurotrophic factors 86:129 glial cell line-derived neurotrophic factors 86:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 3 18 theme conserved 668:676 arg1 points 713:718 both conserved and specificity-determining anchor points 663:718 both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 663:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 1 19 from development 258:268 arg1 system 340:345 the mammalian nervous system 318:345 the mammalian nervous system 318:345 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 5 20 theme functional 986:995 arg1 dissection 997:1006 the functional dissection 982:1006 the functional dissection of the specific versus pleiotropic roles of this system in neurobiology 982:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 5 21 from dissection 997:1006 arg1 neurobiology 1067:1078 neurobiology 1067:1078 neurobiology 1067:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 3 22 theme interaction 604:614 arg1 mode 616:619 a new receptor-ligand interaction mode 582:619 a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 582:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 2 23 theme receptor 435:442 arg1 ARTN 422:425 ARTN 422:425 ARTN 422:425 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 2 23 theme receptor 435:442 arg1 GFRalpha3 444:452 its receptor GFRalpha3 431:452 its receptor GFRalpha3 431:452 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 2 23 theme receptor 435:442 arg1 step 479:482 the initiating step 464:482 the initiating step in the formation of a ternary signaling complex containing the shared RET receptor 464:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 4 24 theme dyad-symmetric 837:850 arg1 interfaces 862:871 dyad-symmetric composite interfaces 837:871 dyad-symmetric composite interfaces 837:871 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 3 25 theme anchor 706:711 arg1 points 713:718 both conserved and specificity-determining anchor points 663:718 both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 663:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 2 26 theme complex 399:405 arg1 structure 382:390 the 1.92 A crystal structure 363:390 the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor 363:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 4 27 with tandem 751:756 arg1 structure 775:783 the complex structure 763:783 the complex structure 763:783 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 0 28 dep recognition 71:81 arg1 Structure 0:8 Structure 0:8 Structure of artemin complexed with its receptor GFRalpha3 0:57 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 5 29 from roles 1043:1047 arg1 neurobiology 1067:1078 neurobiology 1067:1078 neurobiology 1067:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 1 30 theme glial 166:170 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 30 theme glial 166:170 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 3 31 theme GFL-GFRalpha 728:739 arg1 pairs 741:745 all GFL-GFRalpha pairs 724:745 all GFL-GFRalpha pairs 724:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 4 32 theme composite 852:860 arg1 interfaces 862:871 dyad-symmetric composite interfaces 837:871 dyad-symmetric composite interfaces 837:871 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 1 33 theme cell 172:175 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 33 theme cell 172:175 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 0 34 theme artemin 13:19 arg1 Structure 0:8 Structure 0:8 Structure of artemin complexed with its receptor GFRalpha3 0:57 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 0 35 theme factors 123:129 arg1 recognition 71:81 convergent recognition 60:81 convergent recognition of glial cell line-derived neurotrophic factors 60:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 3 36 theme receptor-ligand 588:602 arg1 mode 616:619 a new receptor-ligand interaction mode 582:619 a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 582:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 1 37 theme line-derived 177:188 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 37 theme line-derived 177:188 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 5 38 theme specific 1015:1022 arg1 roles 1043:1047 the specific versus pleiotropic roles 1011:1047 the specific versus pleiotropic roles of this system in neurobiology 1011:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 4 39 theme intracellular 925:937 arg1 signaling 939:947 intracellular signaling 925:947 intracellular signaling 925:947 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 2 40 theme A 372:372 arg1 structure 382:390 the 1.92 A crystal structure 363:390 the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor 363:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 4 41 theme cellular 786:793 arg1 studies 795:801 cellular studies 786:801 cellular studies using receptor chimeras 786:825 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 1 42 theme neurotrophic 190:201 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 42 theme neurotrophic 190:201 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 5 43 theme pleiotropic 1031:1041 arg1 roles 1043:1047 the specific versus pleiotropic roles 1011:1047 the specific versus pleiotropic roles of this system in neurobiology 1011:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 1 44 theme factor 203:208 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 44 theme factor 203:208 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 2 45 theme complex 524:530 arg1 formation 491:499 the formation 487:499 the formation of a ternary signaling complex containing the shared RET receptor 487:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 3 46 theme new 584:586 arg1 mode 616:619 a new receptor-ligand interaction mode 582:619 a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 582:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 2 47 theme initiating 468:477 arg1 ARTN 422:425 ARTN 422:425 ARTN 422:425 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 2 47 theme initiating 468:477 arg1 GFRalpha3 444:452 its receptor GFRalpha3 431:452 its receptor GFRalpha3 431:452 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 2 47 theme initiating 468:477 arg1 step 479:482 the initiating step 464:482 the initiating step in the formation of a ternary signaling complex containing the shared RET receptor 464:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 4 48 theme RET 909:911 arg1 dimerization 893:904 dimerization 893:904 dimerization of RET 893:911 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 4 48 theme RET 909:911 arg1 recruitment 877:887 recruitment 877:887 recruitment 877:887 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 0 49 theme receptor 40:47 arg1 GFRalpha3 49:57 its receptor GFRalpha3 36:57 its receptor GFRalpha3 36:57 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 2 50 theme signaling 514:522 arg1 complex 524:530 a ternary signaling complex 504:530 a ternary signaling complex containing the shared RET receptor 504:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 4 51 theme receptor 809:816 arg1 chimeras 818:825 receptor chimeras 809:825 receptor chimeras 809:825 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 2 52 contain containing 532:541 arg2 receptor 558:565 the shared RET receptor 543:565 the shared RET receptor 543:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 2 52 contain containing 532:541 arg1 complex 524:530 a ternary signaling complex 504:530 a ternary signaling complex containing the shared RET receptor 504:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 1 53 theme many 289:292 arg1 populations 303:313 many neuronal populations 289:313 many neuronal populations 289:313 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 2 54 theme ternary 506:512 arg1 complex 524:530 a ternary signaling complex 504:530 a ternary signaling complex containing the shared RET receptor 504:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 3 55 theme TGF-beta 629:636 arg1 superfamily 638:648 the TGF-beta superfamily 625:648 the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs 625:745 It represents a new receptor-ligand interaction mode for the TGF-beta superfamily that reveals both conserved and specificity-determining anchor points for all GFL-GFRalpha pairs. 16765900 5 56 theme system 1057:1062 arg1 roles 1043:1047 the specific versus pleiotropic roles 1011:1047 the specific versus pleiotropic roles of this system in neurobiology 1011:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 2 57 from step 479:482 arg1 formation 491:499 the formation 487:499 the formation of a ternary signaling complex containing the shared RET receptor 487:565 Here we report the 1.92 A crystal structure of the complex formed between ARTN and its receptor GFRalpha3, which is the initiating step in the formation of a ternary signaling complex containing the shared RET receptor. 16765900 1 58 theme GDNF 211:214 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 58 theme GDNF 211:214 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 0 59 theme convergent 60:69 arg1 recognition 71:81 convergent recognition 60:81 convergent recognition of glial cell line-derived neurotrophic factors 60:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 0 60 link line-derived 97:108 arg1 factors 123:129 glial cell line-derived neurotrophic factors 86:129 glial cell line-derived neurotrophic factors 86:129 Structure of artemin complexed with its receptor GFRalpha3: convergent recognition of glial cell line-derived neurotrophic factors. 16765900 5 61 from neurobiology 1067:1078 arg1 dissection 997:1006 the functional dissection 982:1006 the functional dissection of the specific versus pleiotropic roles of this system in neurobiology 982:1078 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 5 61 from neurobiology 1067:1078 arg1 exploitation 1096:1107 its exploitation 1092:1107 its exploitation for therapeutic applications 1092:1136 These studies should facilitate the functional dissection of the specific versus pleiotropic roles of this system in neurobiology, as well as its exploitation for therapeutic applications. 16765900 1 62 theme neuronal 294:301 arg1 populations 303:313 many neuronal populations 289:313 many neuronal populations 289:313 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 4 63 theme complex 767:773 arg1 structure 775:783 the complex structure 763:783 the complex structure 763:783 In tandem with the complex structure, cellular studies using receptor chimeras implicate dyad-symmetric composite interfaces for recruitment and dimerization of RET, leading to intracellular signaling. 16765900 1 64 theme family 217:222 arg1 GFLs 233:236 GFLs 233:236 GFLs 233:236 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 16765900 1 64 theme family 217:222 arg1 ligands 224:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands 162:230 the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) 162:237 Artemin (ARTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) which regulate the development and maintenance of many neuronal populations in the mammalian nervous system. 12878160 6 0 theme various 840:846 arg1 lines 853:857 various cell lines 840:857 various cell lines 840:857 Using an antiserum prepared against the CRD, the 32-kDa VIPL protein was detected in various cell lines. 12878160 10 1 theme VIPL 1348:1351 arg1 transport 1335:1343 transport 1335:1343 transport of VIPL to the cell surface 1335:1371 Mutating the retrograde transport signal KR to AA resulted in transport of VIPL to the cell surface. 12878160 1 2 theme glycoproteins 135:147 arg1 Subsets 124:130 Subsets 124:130 Subsets of glycoproteins 124:147 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 5 3 theme VIPL 684:687 arg1 mRNA 689:692 The 2.4-kb VIPL mRNA 673:692 The 2.4-kb VIPL mRNA 673:692 The 2.4-kb VIPL mRNA was widely expressed to varying levels in different tissues. 12878160 1 4 theme endoplasmic 235:245 arg1 reticulum 247:255 the endoplasmic reticulum 231:255 the endoplasmic reticulum (ER) 231:260 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 1 4 theme endoplasmic 235:245 arg1 ER 258:259 ER 258:259 ER 258:259 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 9 5 dep ER 1249:1250 arg1 the 1245:1247 the 1245:1247 the 1245:1247 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 10 6 theme signal 1307:1312 arg1 KR 1314:1315 the retrograde transport signal KR 1282:1315 the retrograde transport signal KR 1282:1315 Mutating the retrograde transport signal KR to AA resulted in transport of VIPL to the cell surface. 12878160 7 7 theme plasma 1032:1037 arg1 membrane 1039:1046 the plasma membrane 1028:1046 the plasma membrane 1028:1046 The single N-linked glycan of VIPL remained endoglycosidase H-sensitive during a 2-h pulse-chase, even when the protein was overexpressed or mutated to allow export to the plasma membrane. 12878160 1 8 theme reticulum 247:255 arg1 export 217:222 efficient export out 207:226 efficient export out of the endoplasmic reticulum (ER) 207:260 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 9 9 theme cytoplasmic 1133:1143 arg1 tail 1145:1148 the cytoplasmic tail 1129:1148 the cytoplasmic tail of VIPL 1129:1156 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 7 10 theme VIPL 890:893 arg1 glycan 880:885 The single N-linked glycan 860:885 The single N-linked glycan of VIPL 860:893 The single N-linked glycan of VIPL remained endoglycosidase H-sensitive during a 2-h pulse-chase, even when the protein was overexpressed or mutated to allow export to the plasma membrane. 12878160 11 11 theme VIPL 1397:1400 arg1 mRNA 1402:1405 VIPL mRNA 1397:1405 VIPL mRNA using siRNA 1397:1417 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 3 12 theme related 516:522 arg1 gene 497:500 A gene 495:500 A gene 495:500 A gene, more closely related to VIP36 than to ERGIC-53/p58, and hence called VIPL (VIP36-Like), was identified. 12878160 7 13 theme N-linked 871:878 arg1 glycan 880:885 The single N-linked glycan 860:885 The single N-linked glycan of VIPL 860:893 The single N-linked glycan of VIPL remained endoglycosidase H-sensitive during a 2-h pulse-chase, even when the protein was overexpressed or mutated to allow export to the plasma membrane. 12878160 2 14 dep lectins 341:347 arg1 ERGIC-53/p58 349:360 ERGIC-53/p58 349:360 ERGIC-53/p58 349:360 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 14 dep lectins 341:347 arg1 lectins 341:347 two previously characterized intracellular lectins 298:347 two previously characterized intracellular lectins ERGIC-53/p58 and VIP36 298:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 14 dep lectins 341:347 arg1 VIP36 366:370 VIP36 366:370 VIP36 366:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 11 15 dep glycoproteins 1466:1478 arg1 M 1481:1481 M 1481:1481 M(R) 35 and 250 kDa 1481:1499 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 11 15 dep glycoproteins 1466:1478 arg1 R 1483:1483 R 1483:1483 R 1483:1483 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 5 16 theme 2.4-kb 677:682 arg1 mRNA 689:692 The 2.4-kb VIPL mRNA 673:692 The 2.4-kb VIPL mRNA 673:692 The 2.4-kb VIPL mRNA was widely expressed to varying levels in different tissues. 12878160 9 17 theme characteristic 1199:1212 arg1 KRFY 1185:1188 the sequence KRFY 1172:1188 the sequence KRFY 1172:1188 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 9 17 theme characteristic 1199:1212 arg1 motif 1193:1197 a motif 1191:1197 a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi 1191:1270 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 3 18 dep ERGIC-53/p58 541:552 arg1 to 538:539 to 538:539 to 538:539 A gene, more closely related to VIP36 than to ERGIC-53/p58, and hence called VIPL (VIP36-Like), was identified. 12878160 0 19 theme glycoproteins 78:90 arg1 export 68:73 the export 64:73 the export of glycoproteins from the endoplasmic reticulum 64:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 6 20 theme VIPL 811:814 arg1 protein 816:822 the 32-kDa VIPL protein 800:822 the 32-kDa VIPL protein 800:822 Using an antiserum prepared against the CRD, the 32-kDa VIPL protein was detected in various cell lines. 12878160 7 21 link N-linked 871:878 arg1 glycan 880:885 The single N-linked glycan 860:885 The single N-linked glycan of VIPL 860:893 The single N-linked glycan of VIPL remained endoglycosidase H-sensitive during a 2-h pulse-chase, even when the protein was overexpressed or mutated to allow export to the plasma membrane. 12878160 0 22 from reticulum 113:121 arg1 function 52:59 a putative function 41:59 a putative function in the export of glycoproteins from the endoplasmic reticulum 41:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 0 22 from reticulum 113:121 arg1 export 68:73 the export 64:73 the export of glycoproteins from the endoplasmic reticulum 64:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 11 23 dep M 1481:1481 arg1 kDa 1497:1499 35 and 250 kDa 1486:1499 M(R) 35 and 250 kDa 1481:1499 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 2 24 theme carbohydrate 437:448 arg1 domain 462:467 the conserved carbohydrate recognition domain 423:467 the conserved carbohydrate recognition domain (CRD) 423:473 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 24 theme carbohydrate 437:448 arg1 string 487:492 a search string 478:492 a search string 478:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 24 theme carbohydrate 437:448 arg1 CRD 470:472 CRD 470:472 CRD 470:472 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 6 25 theme 32-kDa 804:809 arg1 protein 816:822 the 32-kDa VIPL protein 800:822 the 32-kDa VIPL protein 800:822 Using an antiserum prepared against the CRD, the 32-kDa VIPL protein was detected in various cell lines. 12878160 7 26 theme single 864:869 arg1 glycan 880:885 The single N-linked glycan 860:885 The single N-linked glycan of VIPL 860:893 The single N-linked glycan of VIPL remained endoglycosidase H-sensitive during a 2-h pulse-chase, even when the protein was overexpressed or mutated to allow export to the plasma membrane. 12878160 2 27 theme conserved 427:435 arg1 domain 462:467 the conserved carbohydrate recognition domain 423:467 the conserved carbohydrate recognition domain (CRD) 423:473 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 27 theme conserved 427:435 arg1 string 487:492 a search string 478:492 a search string 478:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 27 theme conserved 427:435 arg1 CRD 470:472 CRD 470:472 CRD 470:472 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 28 theme related 287:293 arg1 members 279:285 new members 275:285 new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36 275:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 11 29 gly glycoproteins 1466:1478 arg1 glycoproteins 1466:1478 two glycoproteins 1462:1478 two glycoproteins (M(R) 35 and 250 kDa) 1462:1500 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 1 30 theme lectin-like 172:182 arg1 receptors 193:201 lectin-like membrane receptors 172:201 lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER) 172:260 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 0 31 theme VIP36-like 8:17 arg1 protein 28:34 a VIP36-like membrane protein 6:34 a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum 6:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 0 31 theme VIP36-like 8:17 arg1 VIPL 0:3 VIPL 0:3 VIPL 0:3 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 0 32 theme endoplasmic 101:111 arg1 reticulum 113:121 the endoplasmic reticulum 97:121 the endoplasmic reticulum 97:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 0 33 gly glycoproteins 78:90 arg1 glycoproteins 78:90 glycoproteins 78:90 glycoproteins 78:90 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 1 34 theme membrane 184:191 arg1 receptors 193:201 lectin-like membrane receptors 172:201 lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER) 172:260 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 8 35 theme Golgi 1102:1106 arg1 complex 1108:1114 the Golgi complex 1098:1114 the Golgi complex 1098:1114 VIPL localized primarily to the ER and partly to the Golgi complex. 12878160 2 36 theme new 275:277 arg1 members 279:285 new members 275:285 new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36 275:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 37 theme database 401:408 arg1 search 410:415 an extensive database search 388:415 an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string 388:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 38 theme characterized 313:325 arg1 ERGIC-53/p58 349:360 ERGIC-53/p58 349:360 ERGIC-53/p58 349:360 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 38 theme characterized 313:325 arg1 lectins 341:347 two previously characterized intracellular lectins 298:347 two previously characterized intracellular lectins ERGIC-53/p58 and VIP36 298:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 38 theme characterized 313:325 arg1 VIP36 366:370 VIP36 366:370 VIP36 366:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 6 39 located detected 828:835 arg1 lines 853:857 various cell lines 840:857 various cell lines 840:857 Using an antiserum prepared against the CRD, the 32-kDa VIPL protein was detected in various cell lines. 12878160 6 39 located detected 828:835 arg2 protein 816:822 the 32-kDa VIPL protein 800:822 the 32-kDa VIPL protein 800:822 Using an antiserum prepared against the CRD, the 32-kDa VIPL protein was detected in various cell lines. 12878160 2 40 theme extensive 391:399 arg1 search 410:415 an extensive database search 388:415 an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string 388:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 0 41 theme membrane 19:26 arg1 protein 28:34 a VIP36-like membrane protein 6:34 a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum 6:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 0 41 theme membrane 19:26 arg1 VIPL 0:3 VIPL 0:3 VIPL 0:3 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 5 42 theme varying 718:724 arg1 levels 726:731 varying levels 718:731 varying levels 718:731 The 2.4-kb VIPL mRNA was widely expressed to varying levels in different tissues. 12878160 2 43 contain carried 376:382 arg2 search 410:415 an extensive database search 388:415 an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string 388:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 43 contain carried 376:382 arg1 we 373:374 we 373:374 we 373:374 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 1 44 theme efficient 207:215 arg1 export 217:222 efficient export out 207:226 efficient export out of the endoplasmic reticulum (ER) 207:260 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 2 45 theme intracellular 327:339 arg1 ERGIC-53/p58 349:360 ERGIC-53/p58 349:360 ERGIC-53/p58 349:360 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 45 theme intracellular 327:339 arg1 lectins 341:347 two previously characterized intracellular lectins 298:347 two previously characterized intracellular lectins ERGIC-53/p58 and VIP36 298:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 45 theme intracellular 327:339 arg1 VIP36 366:370 VIP36 366:370 VIP36 366:370 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 46 theme recognition 450:460 arg1 domain 462:467 the conserved carbohydrate recognition domain 423:467 the conserved carbohydrate recognition domain (CRD) 423:473 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 46 theme recognition 450:460 arg1 string 487:492 a search string 478:492 a search string 478:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 46 theme recognition 450:460 arg1 CRD 470:472 CRD 470:472 CRD 470:472 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 9 47 theme sequence 1176:1183 arg1 KRFY 1185:1188 the sequence KRFY 1172:1188 the sequence KRFY 1172:1188 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 9 47 theme sequence 1176:1183 arg1 motif 1193:1197 a motif 1191:1197 a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi 1191:1270 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 9 48 theme VIPL 1153:1156 arg1 tail 1145:1148 the cytoplasmic tail 1129:1148 the cytoplasmic tail of VIPL 1129:1156 Like VIP36, the cytoplasmic tail of VIPL terminates in the sequence KRFY, a motif characteristic for proteins recycling between the ER and ERGIC/cis-Golgi. 12878160 10 49 theme cell 1360:1363 arg1 surface 1365:1371 the cell surface 1356:1371 the cell surface 1356:1371 Mutating the retrograde transport signal KR to AA resulted in transport of VIPL to the cell surface. 12878160 11 50 theme glycoproteins 1466:1478 arg1 secretion 1449:1457 the secretion 1445:1457 the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium 1445:1514 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 0 51 from function 52:59 arg1 reticulum 113:121 the endoplasmic reticulum 97:121 the endoplasmic reticulum 97:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 0 51 from function 52:59 arg1 export 68:73 the export 64:73 the export of glycoproteins from the endoplasmic reticulum 64:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 11 52 theme ER 1562:1563 arg1 receptor 1572:1579 an ER export receptor 1559:1579 an ER export receptor 1559:1579 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 11 52 theme ER 1562:1563 arg1 VIPL 1533:1536 VIPL 1533:1536 VIPL 1533:1536 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 10 53 theme transport 1297:1305 arg1 KR 1314:1315 the retrograde transport signal KR 1282:1315 the retrograde transport signal KR 1282:1315 Mutating the retrograde transport signal KR to AA resulted in transport of VIPL to the cell surface. 12878160 1 54 gly glycoproteins 135:147 arg1 glycoproteins 135:147 glycoproteins 135:147 glycoproteins 135:147 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 4 55 from fish 660:663 arg1 evolution 639:647 evolution 639:647 evolution from zebra fish to man 639:670 VIPL has been conserved through evolution from zebra fish to man. 12878160 1 56 theme out 224:226 arg1 export 217:222 efficient export out 207:226 efficient export out of the endoplasmic reticulum (ER) 207:260 Subsets of glycoproteins are thought to require lectin-like membrane receptors for efficient export out of the endoplasmic reticulum (ER). 12878160 2 57 theme search 480:485 arg1 domain 462:467 the conserved carbohydrate recognition domain 423:467 the conserved carbohydrate recognition domain (CRD) 423:473 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 2 57 theme search 480:485 arg1 string 487:492 a search string 478:492 a search string 478:492 To identify new members related to two previously characterized intracellular lectins ERGIC-53/p58 and VIP36, we carried out an extensive database search using the conserved carbohydrate recognition domain (CRD) as a search string. 12878160 10 58 theme retrograde 1286:1295 arg1 KR 1314:1315 the retrograde transport signal KR 1282:1315 the retrograde transport signal KR 1282:1315 Mutating the retrograde transport signal KR to AA resulted in transport of VIPL to the cell surface. 12878160 11 59 theme export 1565:1570 arg1 receptor 1572:1579 an ER export receptor 1559:1579 an ER export receptor 1559:1579 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 11 59 theme export 1565:1570 arg1 VIPL 1533:1536 VIPL 1533:1536 VIPL 1533:1536 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 0 60 theme putative 43:50 arg1 function 52:59 a putative function 41:59 a putative function in the export of glycoproteins from the endoplasmic reticulum 41:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 6 61 theme cell 848:851 arg1 lines 853:857 various cell lines 840:857 various cell lines 840:857 Using an antiserum prepared against the CRD, the 32-kDa VIPL protein was detected in various cell lines. 12878160 0 62 with protein 28:34 arg1 function 52:59 a putative function 41:59 a putative function in the export of glycoproteins from the endoplasmic reticulum 41:121 VIPL, a VIP36-like membrane protein with a putative function in the export of glycoproteins from the endoplasmic reticulum. 12878160 5 63 theme different 736:744 arg1 tissues 746:752 different tissues 736:752 different tissues 736:752 The 2.4-kb VIPL mRNA was widely expressed to varying levels in different tissues. 12878160 11 64 theme mRNA 1402:1405 arg1 knock-down 1383:1392 knock-down 1383:1392 knock-down of VIPL mRNA using siRNA 1383:1417 Finally, knock-down of VIPL mRNA using siRNA significantly slowed down the secretion of two glycoproteins (M(R) 35 and 250 kDa) to the medium, suggesting that VIPL may also function as an ER export receptor. 12878160 4 65 theme zebra 654:658 arg1 fish 660:663 zebra fish 654:663 zebra fish to man 654:670 VIPL has been conserved through evolution from zebra fish to man. 18692472 6 0 dep IL-3 983:986 arg1 receptors 997:1005 receptors 997:1005 receptors 997:1005 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 1 1 theme therapeutic 336:346 arg1 value 348:352 therapeutic value 336:352 therapeutic value for other diseases 336:371 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 0 2 theme receptor 81:88 arg1 activation 90:99 cytokine receptor activation 72:99 cytokine receptor activation 72:99 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 5 3 theme receptor 886:893 arg1 activation 895:904 receptor activation 886:904 receptor activation 886:904 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 4 4 theme GM-CSF/GM-CSF 630:642 arg1 complex 661:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 0 5 theme cytokine 72:79 arg1 activation 90:99 cytokine receptor activation 72:99 cytokine receptor activation 72:99 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 5 6 from interface 805:813 arg1 mutagenesis 753:763 mutagenesis 753:763 mutagenesis of the GM-CSF receptor at the dodecamer interface 753:813 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 5 6 from interface 805:813 arg1 studies 830:836 functional studies 819:836 functional studies 819:836 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 1 7 theme clinical 262:269 arg1 conditions 271:280 clinical conditions 262:280 clinical conditions such as rheumatoid arthritis and leukemia 262:322 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 7 theme clinical 262:269 arg1 leukemia 315:322 leukemia 315:322 leukemia 315:322 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 7 theme clinical 262:269 arg1 arthritis 301:309 rheumatoid arthritis 290:309 rheumatoid arthritis 290:309 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 8 theme other 358:362 arg1 diseases 364:371 other diseases 358:371 other diseases 358:371 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 9 theme pleiotropic 165:175 arg1 factor 144:149 Granulocyte-macrophage colony-stimulating factor 102:149 Granulocyte-macrophage colony-stimulating factor (GM-CSF) 102:158 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 9 theme pleiotropic 165:175 arg1 cytokine 177:184 a pleiotropic cytokine 163:184 a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases 163:371 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 0 10 theme activation 90:99 arg1 mode 64:67 a distinct mode 53:67 a distinct mode of cytokine receptor activation 53:99 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 4 11 theme human 624:628 arg1 complex 661:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 2 12 theme interleukin 495:505 arg1 IL 508:509 the interleukin (IL)-3 491:512 the interleukin (IL)-3 491:512 Its receptors are heterodimers consisting of a ligand-specific alpha subunit and a betac subunit that is shared with the interleukin (IL)-3 and IL-5 receptors. 18692472 4 13 theme higher-order 718:729 arg1 complex 731:737 an unexpected dodecamer or higher-order complex 691:737 complex 731:737 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 4 14 theme ternary 653:659 arg1 complex 661:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 5 15 theme dodecamer 795:803 arg1 interface 805:813 the dodecamer interface 791:813 the dodecamer interface 791:813 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 5 16 theme dodecamer 850:858 arg1 formation 860:868 dodecamer formation 850:868 dodecamer formation 850:868 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 4 17 theme receptor 644:651 arg1 complex 661:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 the human GM-CSF/GM-CSF receptor ternary complex 620:667 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 2 18 dep IL 508:509 arg1 receptors 523:531 receptors 523:531 receptors 523:531 Its receptors are heterodimers consisting of a ligand-specific alpha subunit and a betac subunit that is shared with the interleukin (IL)-3 and IL-5 receptors. 18692472 2 19 theme ligand-specific 421:435 arg1 subunit 443:449 a ligand-specific alpha subunit 419:449 a ligand-specific alpha subunit 419:449 Its receptors are heterodimers consisting of a ligand-specific alpha subunit and a betac subunit that is shared with the interleukin (IL)-3 and IL-5 receptors. 18692472 2 20 theme alpha 437:441 arg1 subunit 443:449 a ligand-specific alpha subunit 419:449 a ligand-specific alpha subunit 419:449 Its receptors are heterodimers consisting of a ligand-specific alpha subunit and a betac subunit that is shared with the interleukin (IL)-3 and IL-5 receptors. 18692472 4 21 theme crystal 599:605 arg1 structure 607:615 the crystal structure 595:615 the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex 595:667 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 0 22 theme receptor 28:35 arg1 complex 37:43 the GM-CSF receptor complex 17:43 the GM-CSF receptor complex 17:43 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 1 23 theme rheumatoid 290:299 arg1 arthritis 301:309 rheumatoid arthritis 290:309 rheumatoid arthritis 290:309 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 0 24 theme GM-CSF 21:26 arg1 complex 37:43 the GM-CSF receptor complex 17:43 the GM-CSF receptor complex 17:43 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 6 25 theme assembly 951:958 arg1 form 934:937 This unusual form 921:937 This unusual form of receptor assembly 921:958 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 5 26 theme GM-CSF 772:777 arg1 receptor 779:786 the GM-CSF receptor 768:786 the GM-CSF receptor 768:786 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 5 27 theme receptor 779:786 arg1 mutagenesis 753:763 mutagenesis 753:763 mutagenesis of the GM-CSF receptor at the dodecamer interface 753:813 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 5 27 theme receptor 779:786 arg1 studies 830:836 functional studies 819:836 functional studies 819:836 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 6 28 theme receptor 942:949 arg1 assembly 951:958 receptor assembly 942:958 receptor assembly 942:958 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 1 29 theme Granulocyte-macrophage 102:123 arg1 factor 144:149 Granulocyte-macrophage colony-stimulating factor 102:149 Granulocyte-macrophage colony-stimulating factor (GM-CSF) 102:158 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 29 theme Granulocyte-macrophage 102:123 arg1 cytokine 177:184 a pleiotropic cytokine 163:184 a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases 163:371 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 29 theme Granulocyte-macrophage 102:123 arg1 GM-CSF 152:157 GM-CSF 152:157 GM-CSF 152:157 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 0 30 theme complex 37:43 arg1 structure 4:12 The structure 0:12 The structure of the GM-CSF receptor complex 0:43 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 2 31 theme betac 457:461 arg1 subunit 463:469 a betac subunit 455:469 a betac subunit that is shared with the interleukin (IL)-3 and IL-5 receptors 455:531 Its receptors are heterodimers consisting of a ligand-specific alpha subunit and a betac subunit that is shared with the interleukin (IL)-3 and IL-5 receptors. 18692472 1 32 theme colony-stimulating 125:142 arg1 factor 144:149 Granulocyte-macrophage colony-stimulating factor 102:149 Granulocyte-macrophage colony-stimulating factor (GM-CSF) 102:158 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 32 theme colony-stimulating 125:142 arg1 cytokine 177:184 a pleiotropic cytokine 163:184 a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases 163:371 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 32 theme colony-stimulating 125:142 arg1 GM-CSF 152:157 GM-CSF 152:157 GM-CSF 152:157 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 6 33 theme therapeutics 1112:1123 arg1 development 1097:1107 the development 1093:1107 the development of therapeutics 1093:1123 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 6 34 theme structural 1020:1029 arg1 basis 1031:1035 a structural basis 1018:1035 a structural basis for understanding their mechanism of activation and for the development of therapeutics 1018:1123 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 4 35 theme unexpected 694:703 arg1 dodecamer 705:713 an unexpected dodecamer or higher-order complex 691:737 dodecamer 705:713 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 1 36 theme blood 231:235 arg1 cells 237:241 blood cells 231:241 blood cells 231:241 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 0 37 theme distinct 55:62 arg1 mode 64:67 a distinct mode 53:67 a distinct mode of cytokine receptor activation 53:99 The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. 18692472 5 38 theme functional 819:828 arg1 studies 830:836 functional studies 819:836 functional studies 819:836 Importantly, mutagenesis of the GM-CSF receptor at the dodecamer interface and functional studies reveal that dodecamer formation is required for receptor activation and signaling. 18692472 6 39 theme activation 1074:1083 arg1 mechanism 1061:1069 their mechanism 1055:1069 their mechanism of activation 1055:1083 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 1 40 theme cells 237:241 arg1 production 204:213 production 204:213 production 204:213 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 1 40 theme cells 237:241 arg1 function 219:226 function 219:226 function 219:226 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 6 41 theme unusual 926:932 arg1 form 934:937 This unusual form 921:937 This unusual form of receptor assembly 921:958 This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors, providing a structural basis for understanding their mechanism of activation and for the development of therapeutics. 18692472 1 42 dep production 204:213 arg1 the 200:202 the 200:202 the 200:202 Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that controls the production and function of blood cells, is deregulated in clinical conditions such as rheumatoid arthritis and leukemia, yet offers therapeutic value for other diseases. 18692472 4 43 theme complex 661:667 arg1 assembly 677:684 its assembly 673:684 its assembly into an unexpected dodecamer or higher-order complex 673:737 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 18692472 4 43 theme complex 661:667 arg1 structure 607:615 the crystal structure 595:615 the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex 595:667 We report here the crystal structure of the human GM-CSF/GM-CSF receptor ternary complex and its assembly into an unexpected dodecamer or higher-order complex. 16754968 2 0 from region 544:549 arg1 stabilization 449:461 stabilization 449:461 stabilization 449:461 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 0 from region 544:549 arg1 position 436:443 position 436:443 position 436:443 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 1 theme binding 163:169 arg1 epitope 171:177 the high-affinity binding epitope 145:177 the high-affinity binding epitope 145:177 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 2 2 theme stabilization 449:461 arg1 indication 418:427 a clearer indication 408:427 a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule 408:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 3 theme high-affinity 149:161 arg1 epitope 171:177 the high-affinity binding epitope 145:177 the high-affinity binding epitope 145:177 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 2 4 dep position 436:443 arg1 the 432:434 the 432:434 the 432:434 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 5 from improvement 317:327 arg1 resolution 332:341 resolution 332:341 resolution 332:341 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 6 theme epitope 171:177 arg1 definition 131:140 better definition 124:140 better definition of the high-affinity binding epitope 124:177 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 2 7 theme electron-density 368:383 arg1 maps 385:388 improved electron-density maps 359:388 improved electron-density maps 359:388 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 8 theme residues 474:481 arg1 stabilization 449:461 stabilization 449:461 stabilization 449:461 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 8 theme residues 474:481 arg1 position 436:443 position 436:443 position 436:443 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 9 dep IL-3 79:82 arg1 signalling 101:110 signalling 101:110 signalling 101:110 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 2 10 theme improved 359:366 arg1 maps 385:388 improved electron-density maps 359:388 improved electron-density maps 359:388 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 11 theme improved 3:10 arg1 structure 23:31 An improved resolution structure 0:31 An improved resolution structure of the human beta common receptor 0:65 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 2 12 from position 436:443 arg1 region 544:549 the elbow region 534:549 the elbow region between domain 1 and domain 4 of the dimer-related molecule 534:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 Tyr15 483:487 Tyr15 483:487 Tyr15 483:487 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 Phe79 490:494 Phe79 490:494 Phe79 490:494 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 Ile350 513:518 Ile350 513:518 Ile350 513:518 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 Tyr403 524:529 Tyr403 524:529 Tyr403 524:529 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 Tyr347 497:502 Tyr347 497:502 Tyr347 497:502 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 His349 505:510 His349 505:510 His349 505:510 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 13 theme key 470:472 arg1 residues 474:481 the key residues 466:481 the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule 466:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 14 theme clearer 410:416 arg1 indication 418:427 a clearer indication 408:427 a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule 408:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 15 theme molecule 602:609 arg1 domain 559:564 domain 1 559:566 domain 1 559:566 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 15 theme molecule 602:609 arg1 domain 572:577 domain 4 572:579 domain 4 of the dimer-related molecule 572:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 15 theme molecule 602:609 arg1 molecule 602:609 the dimer-related molecule 584:609 the dimer-related molecule 584:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 16 theme elbow 538:542 arg1 region 544:549 the elbow region 534:549 the elbow region between domain 1 and domain 4 of the dimer-related molecule 534:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 1 17 theme extracellular 249:261 arg1 domains 263:269 the extracellular domains 245:269 the extracellular domains of the beta common cytokine receptor 245:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 1 17 theme extracellular 249:261 arg1 receptor 299:306 the beta common cytokine receptor 274:306 the beta common cytokine receptor 274:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 2 18 theme dimer-related 588:600 arg1 molecule 602:609 the dimer-related molecule 584:609 the dimer-related molecule 584:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 1 19 theme X-ray 180:184 arg1 diffraction 186:196 X-ray diffraction 180:196 X-ray diffraction 180:196 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 0 20 theme resolution 12:21 arg1 structure 23:31 An improved resolution structure 0:31 An improved resolution structure of the human beta common receptor 0:65 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 1 21 theme domains 263:269 arg1 model 236:240 a model 234:240 a model of the extracellular domains of the beta common cytokine receptor 234:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 2 22 dep residues 474:481 arg1 Tyr15 483:487 Tyr15 483:487 Tyr15 483:487 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 22 dep residues 474:481 arg1 Phe79 490:494 Phe79 490:494 Phe79 490:494 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 22 dep residues 474:481 arg1 Ile350 513:518 Ile350 513:518 Ile350 513:518 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 22 dep residues 474:481 arg1 Tyr403 524:529 Tyr403 524:529 Tyr403 524:529 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 22 dep residues 474:481 arg1 Tyr347 497:502 Tyr347 497:502 Tyr347 497:502 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 22 dep residues 474:481 arg1 His349 505:510 His349 505:510 His349 505:510 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 22 dep residues 474:481 arg1 residues 474:481 the key residues 466:481 the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule 466:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 23 from stabilization 449:461 arg1 region 544:549 the elbow region 534:549 the elbow region between domain 1 and domain 4 of the dimer-related molecule 534:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 2 24 theme position 436:443 arg1 indication 418:427 a clearer indication 408:427 a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule 408:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 25 theme beta 46:49 arg1 receptor 58:65 the human beta common receptor 36:65 the human beta common receptor 36:65 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 1 26 theme beta 278:281 arg1 receptor 299:306 the beta common cytokine receptor 274:306 the beta common cytokine receptor 274:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 2 27 theme minor 311:315 arg1 improvement 317:327 A minor improvement 309:327 A minor improvement in resolution 309:341 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 28 theme human 40:44 arg1 receptor 58:65 the human beta common receptor 36:65 the human beta common receptor 36:65 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 1 29 theme common 283:288 arg1 receptor 299:306 the beta common cytokine receptor 274:306 the beta common cytokine receptor 274:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 0 30 theme better 124:129 arg1 definition 131:140 better definition 124:140 better definition of the high-affinity binding epitope 124:177 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 1 31 theme cytokine 290:297 arg1 receptor 299:306 the beta common cytokine receptor 274:306 the beta common cytokine receptor 274:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 2 32 from residues 474:481 arg1 region 544:549 the elbow region 534:549 the elbow region between domain 1 and domain 4 of the dimer-related molecule 534:609 A minor improvement in resolution has resulted in improved electron-density maps, which have given a clearer indication of the position and stabilization of the key residues Tyr15, Phe79, Tyr347, His349, Ile350 and Tyr403 in the elbow region between domain 1 and domain 4 of the dimer-related molecule. 16754968 0 33 theme receptor 58:65 arg1 structure 23:31 An improved resolution structure 0:31 An improved resolution structure of the human beta common receptor 0:65 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 1 34 used used 207:210 arg2 diffraction 186:196 X-ray diffraction 180:196 X-ray diffraction 180:196 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 0 35 theme common 51:56 arg1 receptor 58:65 the human beta common receptor 36:65 the human beta common receptor 36:65 An improved resolution structure of the human beta common receptor involved in IL-3, IL-5 and GM-CSF signalling which gives better definition of the high-affinity binding epitope. 16754968 1 36 theme receptor 299:306 arg1 domains 263:269 the extracellular domains 245:269 the extracellular domains of the beta common cytokine receptor 245:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 16754968 1 36 theme receptor 299:306 arg1 receptor 299:306 the beta common cytokine receptor 274:306 the beta common cytokine receptor 274:306 X-ray diffraction has been used to produce and refine a model of the extracellular domains of the beta common cytokine receptor. 21959264 11 0 theme reported 1425:1432 arg1 TLR2/TLR6 1478:1486 TLR2/TLR6 1478:1486 TLR2/TLR6 1478:1486 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 0 theme reported 1425:1432 arg1 TLR3 1493:1496 TLR3 1493:1496 TLR3 1493:1496 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 0 theme reported 1425:1432 arg1 TLR1/TLR2 1467:1475 TLR1/TLR2 1467:1475 TLR1/TLR2 1467:1475 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 0 theme reported 1425:1432 arg1 dimers 1438:1443 previously reported TLR dimers 1414:1443 previously reported TLR dimers 1414:1443 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 0 theme reported 1425:1432 arg1 TLR4/MD-2 1456:1464 TLR4/MD-2 1456:1464 TLR4/MD-2 1456:1464 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 6 1 theme oligomer 830:837 arg1 structure 783:791 the structure 779:791 the structure of the RP105/MD-1 heterodimer or its oligomer 779:837 Little is known, however, about the structure of the RP105/MD-1 heterodimer or its oligomer. 21959264 7 2 theme mouse 891:895 arg1 complexes 918:926 mouse and human RP105/MD-1 complexes 891:926 mouse and human RP105/MD-1 complexes 891:926 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 21959264 6 3 theme heterodimer 811:821 arg1 structure 783:791 the structure 779:791 the structure of the RP105/MD-1 heterodimer or its oligomer 779:837 Little is known, however, about the structure of the RP105/MD-1 heterodimer or its oligomer. 21959264 0 4 theme dimer 73:77 arg1 organization 79:90 unique dimer organization 66:90 unique dimer organization of the toll-like receptor family 66:123 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 10 5 theme TLR4/MD-2 1320:1328 arg1 complex 1330:1336 the 2:2 TLR4/MD-2 complex 1312:1336 the 2:2 TLR4/MD-2 complex 1312:1336 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 7 6 theme crystal 869:875 arg1 structures 877:886 the crystal structures 865:886 the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions 865:955 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 21959264 9 7 theme inverse 1142:1148 arg1 arrangement 1150:1160 an inverse arrangement 1139:1160 an inverse arrangement 1139:1160 The "m"-shaped 2:2 RP105/MD-1 complex exhibits an inverse arrangement, with N-termini interacting in the middle. 21959264 11 8 theme cytoplasmic 1578:1588 arg1 domain 1600:1605 the C-terminal cytoplasmic signaling domain 1563:1605 the C-terminal cytoplasmic signaling domain 1563:1605 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 9 9 theme -shaped 1099:1105 arg1 complex 1122:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex exhibits an inverse arrangement, with N-termini interacting in the middle. 21959264 10 10 theme 2:2 1316:1318 arg1 complex 1330:1336 the 2:2 TLR4/MD-2 complex 1312:1336 the 2:2 TLR4/MD-2 complex 1312:1336 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 5 11 contain has 574:576 arg2 cavity 592:597 a hydrophobic cavity 578:597 a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products 578:744 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 5 11 contain has 574:576 arg1 MD-1 564:567 MD-1 564:567 MD-1 564:567 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 11 12 theme heterotypic 1536:1546 arg1 interaction 1548:1558 homotypic or heterotypic interaction 1523:1558 homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain 1523:1605 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 5 13 theme lipid 643:647 arg1 A 649:649 lipid A 643:649 lipid A 643:649 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 5 14 theme lipid 617:621 arg1 IVa 623:625 lipid IVa 617:625 lipid IVa 617:625 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 5 14 theme lipid 617:621 arg1 precursor 630:638 a precursor 628:638 a precursor of lipid A 628:649 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 0 15 theme receptor 109:116 arg1 family 118:123 the toll-like receptor family 95:123 the toll-like receptor family 95:123 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 6 16 theme RP105/MD-1 800:809 arg1 heterodimer 811:821 the RP105/MD-1 heterodimer 796:821 the RP105/MD-1 heterodimer 796:821 Little is known, however, about the structure of the RP105/MD-1 heterodimer or its oligomer. 21959264 10 17 located located 1255:1261 arg2 located 1255:1261 located 1255:1261 located 1255:1261 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 10 17 located located 1255:1261 arg1 side 1279:1282 the opposite side 1266:1282 the opposite side of the complex 1266:1297 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 10 17 located located 1255:1261 arg2 interface 1228:1236 the dimerization interface 1211:1236 the dimerization interface of RP105/MD-1 1211:1250 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 3 18 theme lipid 435:439 arg1 A 441:441 lipid A 435:441 lipid A 435:441 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 3 18 theme lipid 435:439 arg1 center 455:460 the active center 444:460 the active center of LPS 444:467 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 4 19 with interface 505:513 arg1 TLR4 520:523 TLR4 520:523 TLR4 520:523 LPS-bound MD-2 opens the secondary interface with TLR4, leading to dimerization of TLR4/MD-2. 21959264 1 20 theme 4/MD-2 155:160 arg1 heterodimer 162:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer 126:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer 126:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer senses lipopolysaccharide (LPS). 21959264 0 21 theme toll-like 99:107 arg1 family 118:123 the toll-like receptor family 95:123 the toll-like receptor family 95:123 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 5 22 theme sensing 704:710 arg1 LPS 712:714 sensing LPS 704:714 sensing LPS 704:714 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 2 23 theme TLR-related 242:252 arg1 RP105 207:211 RP105 207:211 RP105 (radioprotective 105 kDa) 207:237 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 2 23 theme TLR-related 242:252 arg1 molecule 254:261 a TLR-related molecule 240:261 a TLR-related molecule 240:261 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 2 24 theme MD-2-like 355:363 arg1 molecule 365:372 the MD-2-like molecule 351:372 the MD-2-like molecule 351:372 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 2 24 theme MD-2-like 355:363 arg1 MD-1 345:348 MD-1 345:348 MD-1 345:348 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 5 25 theme microbial 727:735 arg1 products 737:744 related microbial products 719:744 related microbial products 719:744 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 8 26 theme complex 1047:1053 arg1 dimerization 1012:1023 dimerization 1012:1023 dimerization of the 1:1 RP105/MD-1 complex 1012:1053 Both mouse and human RP105/MD-1 exhibit dimerization of the 1:1 RP105/MD-1 complex, demonstrating a novel organization. 21959264 8 27 theme RP105/MD-1 1036:1045 arg1 complex 1047:1053 the 1:1 RP105/MD-1 complex 1028:1053 the 1:1 RP105/MD-1 complex 1028:1053 Both mouse and human RP105/MD-1 exhibit dimerization of the 1:1 RP105/MD-1 complex, demonstrating a novel organization. 21959264 3 28 theme unique 386:391 arg1 cavity 405:410 a unique hydrophobic cavity 384:410 a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS 384:467 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 0 29 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of mouse and human RP105/MD-1 complexes 0:57 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 8 30 theme novel 1072:1076 arg1 organization 1078:1089 a novel organization 1070:1089 a novel organization 1070:1089 Both mouse and human RP105/MD-1 exhibit dimerization of the 1:1 RP105/MD-1 complex, demonstrating a novel organization. 21959264 5 31 theme hydrophobic 580:590 arg1 cavity 592:597 a hydrophobic cavity 578:597 a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products 578:744 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 9 32 theme 2:2 1107:1109 arg1 complex 1122:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex exhibits an inverse arrangement, with N-termini interacting in the middle. 21959264 3 33 theme hydrophobic 393:403 arg1 cavity 405:410 a unique hydrophobic cavity 384:410 a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS 384:467 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 0 34 theme mouse 22:26 arg1 complexes 49:57 mouse and human RP105/MD-1 complexes 22:57 mouse and human RP105/MD-1 complexes 22:57 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 4 35 theme secondary 495:503 arg1 interface 505:513 the secondary interface 491:513 the secondary interface with TLR4 491:523 LPS-bound MD-2 opens the secondary interface with TLR4, leading to dimerization of TLR4/MD-2. 21959264 3 36 contain has 380:382 arg1 MD-2 375:378 MD-2 375:378 MD-2 375:378 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 3 36 contain has 380:382 arg2 cavity 405:410 a unique hydrophobic cavity 384:410 a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS 384:467 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 11 37 theme TLR 1434:1436 arg1 TLR2/TLR6 1478:1486 TLR2/TLR6 1478:1486 TLR2/TLR6 1478:1486 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 37 theme TLR 1434:1436 arg1 TLR3 1493:1496 TLR3 1493:1496 TLR3 1493:1496 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 37 theme TLR 1434:1436 arg1 TLR1/TLR2 1467:1475 TLR1/TLR2 1467:1475 TLR1/TLR2 1467:1475 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 37 theme TLR 1434:1436 arg1 dimers 1438:1443 previously reported TLR dimers 1414:1443 previously reported TLR dimers 1414:1443 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 37 theme TLR 1434:1436 arg1 TLR4/MD-2 1456:1464 TLR4/MD-2 1456:1464 TLR4/MD-2 1456:1464 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 10 38 theme complex 1291:1297 arg1 side 1279:1282 the opposite side 1266:1282 the opposite side of the complex 1266:1297 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 0 39 theme family 118:123 arg1 organization 79:90 unique dimer organization 66:90 unique dimer organization of the toll-like receptor family 66:123 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 5 40 from role 665:668 arg1 products 737:744 related microbial products 719:744 related microbial products 719:744 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 5 40 from role 665:668 arg1 LPS 712:714 sensing LPS 704:714 sensing LPS 704:714 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 11 41 from dimers 1438:1443 arg1 distinct 1400:1407 distinct 1400:1407 distinct 1400:1407 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 41 from dimers 1438:1443 arg1 complex 1389:1395 the 2:2 RP105/MD-1 complex 1370:1395 the 2:2 RP105/MD-1 complex 1370:1395 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 2 42 dep RP105 207:211 arg1 radioprotective 214:228 radioprotective 214:228 radioprotective 214:228 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 0 43 theme human 32:36 arg1 complexes 49:57 mouse and human RP105/MD-1 complexes 22:57 mouse and human RP105/MD-1 complexes 22:57 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 3 44 theme active 448:453 arg1 A 441:441 lipid A 435:441 lipid A 435:441 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 3 44 theme active 448:453 arg1 center 455:460 the active center 444:460 the active center of LPS 444:467 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 5 45 theme A 649:649 arg1 IVa 623:625 lipid IVa 617:625 lipid IVa 617:625 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 5 45 theme A 649:649 arg1 precursor 630:638 a precursor 628:638 a precursor of lipid A 628:649 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 11 46 theme signaling 1590:1598 arg1 domain 1600:1605 the C-terminal cytoplasmic signaling domain 1563:1605 the C-terminal cytoplasmic signaling domain 1563:1605 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 8 47 theme human 987:991 arg1 RP105/MD-1 993:1002 human RP105/MD-1 987:1002 human RP105/MD-1 987:1002 Both mouse and human RP105/MD-1 exhibit dimerization of the 1:1 RP105/MD-1 complex, demonstrating a novel organization. 21959264 10 48 theme RP105/MD-1 1241:1250 arg1 located 1255:1261 located 1255:1261 located 1255:1261 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 10 48 theme RP105/MD-1 1241:1250 arg1 interface 1228:1236 the dimerization interface 1211:1236 the dimerization interface of RP105/MD-1 1211:1250 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 2 49 dep radioprotective 214:228 arg1 kDa 234:236 105 kDa 230:236 105 kDa 230:236 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 0 50 theme complexes 49:57 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of mouse and human RP105/MD-1 complexes 0:57 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 4 51 theme LPS-bound 470:478 arg1 MD-2 480:483 LPS-bound MD-2 470:483 LPS-bound MD-2 470:483 LPS-bound MD-2 opens the secondary interface with TLR4, leading to dimerization of TLR4/MD-2. 21959264 5 52 theme related 719:725 arg1 products 737:744 related microbial products 719:744 related microbial products 719:744 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 8 53 theme 1:1 1032:1034 arg1 complex 1047:1053 the 1:1 RP105/MD-1 complex 1028:1053 the 1:1 RP105/MD-1 complex 1028:1053 Both mouse and human RP105/MD-1 exhibit dimerization of the 1:1 RP105/MD-1 complex, demonstrating a novel organization. 21959264 2 54 theme leucine-rich 309:320 arg1 repeats 322:328 the extracellular leucine-rich repeats 291:328 the extracellular leucine-rich repeats 291:328 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 0 55 theme RP105/MD-1 38:47 arg1 complexes 49:57 mouse and human RP105/MD-1 complexes 22:57 mouse and human RP105/MD-1 complexes 22:57 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 7 56 theme RP105/MD-1 907:916 arg1 complexes 918:926 mouse and human RP105/MD-1 complexes 891:926 mouse and human RP105/MD-1 complexes 891:926 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 21959264 2 57 theme extracellular 295:307 arg1 repeats 322:328 the extracellular leucine-rich repeats 291:328 the extracellular leucine-rich repeats 291:328 RP105 (radioprotective 105 kDa), a TLR-related molecule, is similar to TLR4 in that the extracellular leucine-rich repeats associate with MD-1, the MD-2-like molecule. 21959264 11 58 theme 2:2 1374:1376 arg1 distinct 1400:1407 distinct 1400:1407 distinct 1400:1407 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 58 theme 2:2 1374:1376 arg1 complex 1389:1395 the 2:2 RP105/MD-1 complex 1370:1395 the 2:2 RP105/MD-1 complex 1370:1395 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 9 59 theme RP105/MD-1 1111:1120 arg1 complex 1122:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex exhibits an inverse arrangement, with N-termini interacting in the middle. 21959264 1 60 theme receptor 140:147 arg1 heterodimer 162:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer 126:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer 126:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer senses lipopolysaccharide (LPS). 21959264 7 61 theme complexes 918:926 arg1 structures 877:886 the crystal structures 865:886 the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions 865:955 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 21959264 11 62 theme homotypic 1523:1531 arg1 interaction 1548:1558 homotypic or heterotypic interaction 1523:1558 homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain 1523:1605 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 63 theme RP105/MD-1 1378:1387 arg1 distinct 1400:1407 distinct 1400:1407 distinct 1400:1407 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 11 63 theme RP105/MD-1 1378:1387 arg1 complex 1389:1395 the 2:2 RP105/MD-1 complex 1370:1395 the 2:2 RP105/MD-1 complex 1370:1395 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 4 64 theme TLR4/MD-2 553:561 arg1 dimerization 537:548 dimerization 537:548 dimerization of TLR4/MD-2 537:561 LPS-bound MD-2 opens the secondary interface with TLR4, leading to dimerization of TLR4/MD-2. 21959264 10 65 theme dimerization 1215:1226 arg1 located 1255:1261 located 1255:1261 located 1255:1261 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 10 65 theme dimerization 1215:1226 arg1 interface 1228:1236 the dimerization interface 1211:1236 the dimerization interface of RP105/MD-1 1211:1250 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 11 66 theme domain 1600:1605 arg1 interaction 1548:1558 homotypic or heterotypic interaction 1523:1558 homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain 1523:1605 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 1 67 theme Toll-like 130:138 arg1 TLR 150:152 TLR 150:152 TLR 150:152 The Toll-like receptor (TLR) 4/MD-2 heterodimer senses lipopolysaccharide (LPS). 21959264 1 67 theme Toll-like 130:138 arg1 receptor 140:147 Toll-like receptor 130:147 The Toll-like receptor (TLR) 4/MD-2 heterodimer 126:172 The Toll-like receptor (TLR) 4/MD-2 heterodimer senses lipopolysaccharide (LPS). 21959264 9 68 theme m 1097:1097 arg1 complex 1122:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex 1092:1128 The "m"-shaped 2:2 RP105/MD-1 complex exhibits an inverse arrangement, with N-termini interacting in the middle. 21959264 0 69 theme unique 66:71 arg1 organization 79:90 unique dimer organization 66:90 unique dimer organization of the toll-like receptor family 66:123 Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. 21959264 7 70 theme Å 943:943 arg1 resolutions 945:955 1.9 and 2.8 Å resolutions 931:955 1.9 and 2.8 Å resolutions 931:955 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 21959264 7 71 theme human 901:905 arg1 complexes 918:926 mouse and human RP105/MD-1 complexes 891:926 mouse and human RP105/MD-1 complexes 891:926 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 21959264 3 72 theme LPS 465:467 arg1 A 441:441 lipid A 435:441 lipid A 435:441 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 3 72 theme LPS 465:467 arg1 center 455:460 the active center 444:460 the active center of LPS 444:467 MD-2 has a unique hydrophobic cavity that directly binds to lipid A, the active center of LPS. 21959264 10 73 theme opposite 1270:1277 arg1 side 1279:1282 the opposite side 1266:1282 the opposite side of the complex 1266:1297 Thus, the dimerization interface of RP105/MD-1 is located on the opposite side of the complex, compared to the 2:2 TLR4/MD-2 complex. 21959264 5 74 theme RP105/MD-1 678:687 arg1 heterodimer 689:699 the RP105/MD-1 heterodimer 674:699 the RP105/MD-1 heterodimer 674:699 MD-1 also has a hydrophobic cavity that accommodates lipid IVa, a precursor of lipid A, suggesting a role for the RP105/MD-1 heterodimer in sensing LPS or related microbial products. 21959264 11 75 theme C-terminal 1567:1576 arg1 domain 1600:1605 the C-terminal cytoplasmic signaling domain 1563:1605 the C-terminal cytoplasmic signaling domain 1563:1605 These results demonstrate that the 2:2 RP105/MD-1 complex is distinct from previously reported TLR dimers, including TLR4/MD-2, TLR1/TLR2, TLR2/TLR6, and TLR3, all of which facilitate homotypic or heterotypic interaction of the C-terminal cytoplasmic signaling domain. 21959264 7 76 from resolutions 945:955 arg1 structures 877:886 the crystal structures 865:886 the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions 865:955 Here, we have determined the crystal structures of mouse and human RP105/MD-1 complexes at 1.9 and 2.8 Å resolutions, respectively. 2137200 3 0 theme interleukin-1 335:347 arg1 receptor 349:356 the interleukin-1 receptor 331:356 the interleukin-1 receptor 331:356 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 1 1 theme human 171:175 arg1 monocytes 177:185 human monocytes 171:185 human monocytes 171:185 Three interleukin-1 inhibitors have been purified to homogeneity from medium conditioned by human monocytes. 2137200 3 2 theme high 411:414 arg1 concentrations 416:429 very high concentrations 406:429 very high concentrations 406:429 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 3 3 dep binds 322:326 arg1 even 398:401 even 398:401 even 398:401 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 2 4 theme protein 301:307 arg1 these 259:263 these 259:263 these 259:263 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 2 4 theme protein 301:307 arg1 forms 283:287 glycosylation forms 269:287 glycosylation forms of a single protein 269:307 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 3 5 theme pure 451:454 arg1 antagonist 465:474 a pure receptor antagonist 449:474 a pure receptor antagonist 449:474 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 3 6 theme interleukin-1-like 369:386 arg1 activity 388:395 no interleukin-1-like activity 366:395 no interleukin-1-like activity 366:395 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 1 7 theme interleukin-1 85:97 arg1 inhibitors 99:108 Three interleukin-1 inhibitors 79:108 Three interleukin-1 inhibitors 79:108 Three interleukin-1 inhibitors have been purified to homogeneity from medium conditioned by human monocytes. 2137200 3 8 theme receptor 456:463 arg1 antagonist 465:474 a pure receptor antagonist 449:474 a pure receptor antagonist 449:474 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 0 9 theme receptor 14:21 arg1 activity 34:41 Interleukin-1 receptor antagonist activity 0:41 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 0:77 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 2137200 0 10 theme Interleukin-1 0:12 arg1 activity 34:41 Interleukin-1 receptor antagonist activity 0:41 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 0:77 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 2137200 0 11 theme antagonist 23:32 arg1 activity 34:41 Interleukin-1 receptor antagonist activity 0:41 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 0:77 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 2137200 2 12 theme sequence 196:203 arg1 analysis 205:212 Partial sequence analysis 188:212 Partial sequence analysis 188:212 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 2 13 theme Partial 188:194 arg1 analysis 205:212 Partial sequence analysis 188:212 Partial sequence analysis 188:212 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 0 14 theme interleukin-1 54:66 arg1 inhibitor 68:76 a human interleukin-1 inhibitor 46:76 a human interleukin-1 inhibitor 46:76 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 2137200 0 15 theme human 48:52 arg1 inhibitor 68:76 a human interleukin-1 inhibitor 46:76 a human interleukin-1 inhibitor 46:76 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 2137200 2 16 with analysis 205:212 arg1 N-glycanase 233:243 N-glycanase 233:243 N-glycanase 233:243 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 2 17 theme single 294:299 arg1 protein 301:307 a single protein 292:307 a single protein 292:307 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 2 18 with digestion 218:226 arg1 N-glycanase 233:243 N-glycanase 233:243 N-glycanase 233:243 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 0 19 theme inhibitor 68:76 arg1 activity 34:41 Interleukin-1 receptor antagonist activity 0:41 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 0:77 Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. 2137200 2 20 theme glycosylation 269:281 arg1 these 259:263 these 259:263 these 259:263 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 2 20 theme glycosylation 269:281 arg1 forms 283:287 glycosylation forms 269:287 glycosylation forms of a single protein 269:307 Partial sequence analysis and digestion with N-glycanase indicate that these are glycosylation forms of a single protein. 2137200 3 21 contain has 362:364 arg2 activity 388:395 no interleukin-1-like activity 366:395 no interleukin-1-like activity 366:395 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 2137200 3 21 contain has 362:364 arg1 protein 314:320 The protein 310:320 The protein 310:320 The protein binds to the interleukin-1 receptor but has no interleukin-1-like activity, even at very high concentrations, and is therefore a pure receptor antagonist. 8670172 3 0 theme mass 718:721 arg1 MALDI-MS 737:744 MALDI-MS 737:744 MALDI-MS 737:744 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 0 theme mass 718:721 arg1 spectrometry 723:734 matrix-assisted laser-desorption mass spectrometry 685:734 matrix-assisted laser-desorption mass spectrometry (MALDI-MS) 685:745 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 7 1 theme anchor 1136:1141 arg1 forms 1123:1127 Variant forms 1115:1127 Variant forms of the anchor 1115:1141 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 10 2 theme oligosaccharide 1425:1439 arg1 chains 1441:1446 The predominant oligosaccharide chains 1409:1446 The predominant oligosaccharide chains 1409:1446 The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. 8670172 11 3 theme terminal 1651:1658 arg1 complexes 1678:1686 the terminal complement SC5b-8 complexes 1647:1686 the terminal complement SC5b-8 complexes 1647:1686 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 10 4 theme triantennary 1481:1492 arg1 complexes 1494:1502 biantennary and triantennary complexes 1465:1502 biantennary and triantennary complexes 1465:1502 The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. 8670172 7 5 theme units 1205:1209 arg1 deletions 1147:1155 deletions 1147:1155 deletions 1147:1155 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 14 6 theme CD59 2020:2023 arg1 activity 2008:2015 the full functional activity 1988:2015 the full functional activity of CD59 1988:2023 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 8 7 theme Sialic 1212:1217 arg1 acid 1219:1222 Sialic acid 1212:1222 Sialic acid linked to an N-acetylhexosamine-galactose arm 1212:1268 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 14 8 theme CD59 1926:1929 arg1 anchor 1887:1892 the GPI anchor 1879:1892 the GPI anchor 1879:1892 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 14 8 theme CD59 1926:1929 arg1 oligosaccharide 1907:1921 N-linked oligosaccharide 1898:1921 N-linked oligosaccharide 1898:1921 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 7 9 theme Variant 1115:1121 arg1 forms 1123:1127 Variant forms 1115:1127 Variant forms of the anchor 1115:1141 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 10 10 theme biantennary 1465:1475 arg1 complexes 1494:1502 biantennary and triantennary complexes 1465:1502 biantennary and triantennary complexes 1465:1502 The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. 8670172 3 11 theme CD59 578:581 arg1 composition 523:533 the detailed structural composition 499:533 the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) 499:589 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 15 12 theme proteins 2196:2203 arg1 solubilization 2165:2178 the solubilization 2161:2178 the solubilization of GPI-anchored proteins 2161:2203 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 0 13 theme GPI 135:137 arg1 anchor 140:145 glycophosphoinositol (GPI) anchor 113:145 glycophosphoinositol (GPI) anchor 113:145 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 3 14 theme soluble 562:568 arg1 CD59u 584:588 CD59u 584:588 CD59u 584:588 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 14 theme soluble 562:568 arg1 CD59 578:581 the naturally occurring soluble urinary CD59 538:581 the naturally occurring soluble urinary CD59 (CD59u) 538:589 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 5 15 theme peptide 843:849 arg1 analysis 807:814 Mass analysis 802:814 Mass analysis of the isolated C-terminal peptide (T9) 802:854 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 13 16 dep lower 1779:1783 arg1 efficiency 1785:1794 efficiency 1785:1794 efficiency 1785:1794 200-fold lower efficiency than erythrocyte CD59. 8670172 2 17 from fluids 470:475 arg1 present 448:454 present 448:454 present 448:454 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 0 18 theme anchor 140:145 arg1 heterogeneity 72:84 heterogeneity 72:84 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 0:203 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 5 19 attach present 944:950 arg2 GPI-anchor 873:882 a GPI-anchor 871:882 a GPI-anchor (at Asn-77) without an inositol-associated phospholipid 871:938 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 19 attach present 944:950 arg2 present 944:950 present 944:950 present 944:950 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 19 attach present 944:950 arg1 CD59u 963:967 soluble CD59u 955:967 soluble CD59u 955:967 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 0 20 theme mass 176:179 arg1 analysis 195:202 laser-desorption mass spectrometric analysis 159:202 laser-desorption mass spectrometric analysis 159:202 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 3 21 theme anion-exchange 614:627 arg1 chromatography 629:642 anion-exchange chromatography 614:642 anion-exchange chromatography 614:642 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 5 22 theme inositol-associated 907:925 arg1 phospholipid 927:938 an inositol-associated phospholipid 904:938 an inositol-associated phospholipid 904:938 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 14 23 gly heterogeneity 1857:1869 arg1 anchor 1887:1892 the GPI anchor 1879:1892 the GPI anchor 1879:1892 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 14 23 gly heterogeneity 1857:1869 arg1 oligosaccharide 1907:1921 N-linked oligosaccharide 1898:1921 N-linked oligosaccharide 1898:1921 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 9 24 theme N-linked 1312:1319 arg1 chain 1339:1343 The N-linked carbohydrate side chain 1308:1343 The N-linked carbohydrate side chain of CD59u (at Asn-18) 1308:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 15 25 dep phosphate 2075:2083 arg1 the 2055:2057 the 2055:2057 the 2055:2057 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 6 26 theme chemical 1013:1020 arg1 modification 1022:1033 chemical modification 1013:1033 chemical modification 1013:1033 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 14 27 theme oligosaccharide 1907:1921 arg1 heterogeneity 1857:1869 the structural heterogeneity 1842:1869 the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 1842:1929 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 15 28 theme diradylglycerol 2059:2073 arg1 phosphate 2075:2083 diradylglycerol phosphate 2059:2083 diradylglycerol phosphate 2059:2083 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 9 29 theme side 1334:1337 arg1 chain 1339:1343 The N-linked carbohydrate side chain 1308:1343 The N-linked carbohydrate side chain of CD59u (at Asn-18) 1308:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 8 30 located found 1274:1278 arg1 variants 1298:1305 two GPI-anchor variants 1283:1305 two GPI-anchor variants 1283:1305 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 8 30 located found 1274:1278 arg2 acid 1219:1222 Sialic acid 1212:1222 Sialic acid linked to an N-acetylhexosamine-galactose arm 1212:1268 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 7 31 contain had 1143:1145 arg1 forms 1123:1127 Variant forms 1115:1127 Variant forms of the anchor 1115:1141 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 7 31 contain had 1143:1145 arg2 deletions 1147:1155 deletions 1147:1155 deletions 1147:1155 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 7 31 contain had 1143:1145 arg2 extensions 1164:1173 extensions 1164:1173 extensions 1164:1173 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 0 32 theme oligosaccharide 93:107 arg1 heterogeneity 72:84 heterogeneity 72:84 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 0:203 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 1 33 theme epithelial 355:364 arg1 cells 366:370 epithelial cells 355:370 epithelial cells 355:370 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 2 34 attach present 448:454 arg2 forms 430:434 soluble lipid-free forms 411:434 soluble lipid-free forms of CD59 411:442 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 2 34 attach present 448:454 arg1 fluids 470:475 human body fluids 459:475 human body fluids 459:475 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 0 35 gly heterogeneity 72:84 arg1 anchor 140:145 glycophosphoinositol (GPI) anchor 113:145 glycophosphoinositol (GPI) anchor 113:145 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 0 35 gly heterogeneity 72:84 arg1 oligosaccharide 93:107 oligosaccharide 93:107 oligosaccharide 93:107 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 14 36 theme phospholipid 1956:1967 arg1 tail 1969:1972 the phospholipid tail 1952:1972 the phospholipid tail 1952:1972 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 2 37 from present 448:454 arg1 fluids 470:475 human body fluids 459:475 human body fluids 459:475 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 4 38 theme 12444 783:787 arg1 M 775:775 an average M 764:775 an average M(r) of 12444 in MALDI-MS 764:799 CD59u exhibited an average M(r) of 12444 in MALDI-MS. 8670172 4 38 theme 12444 783:787 arg1 r 777:777 r 777:777 r 777:777 CD59u exhibited an average M(r) of 12444 in MALDI-MS. 8670172 2 39 theme lipid-tailed 392:403 arg1 CD59 405:408 the lipid-tailed CD59 388:408 the lipid-tailed CD59 388:408 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 0 40 theme glycophosphoinositol 113:132 arg1 anchor 140:145 glycophosphoinositol (GPI) anchor 113:145 glycophosphoinositol (GPI) anchor 113:145 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 0 41 dep oligosaccharide 93:107 arg1 the 89:91 the 89:91 the 89:91 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 4 42 theme average 767:773 arg1 M 775:775 an average M 764:775 an average M(r) of 12444 in MALDI-MS 764:799 CD59u exhibited an average M(r) of 12444 in MALDI-MS. 8670172 4 42 theme average 767:773 arg1 r 777:777 r 777:777 r 777:777 CD59u exhibited an average M(r) of 12444 in MALDI-MS. 8670172 0 43 theme Structural 0:9 arg1 composition 11:21 Structural composition 0:21 Structural composition 0:21 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 1 44 theme membrane 280:287 arg1 complex 296:302 the membrane attack complex 276:302 the membrane attack complex of complement 276:316 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 6 45 theme GPI-anchor 1078:1087 arg1 variants 1089:1096 seven different GPI-anchor variants 1062:1096 seven different GPI-anchor variants 1062:1096 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 10 46 gly fucosylated 1453:1463 arg1 chains 1441:1446 The predominant oligosaccharide chains 1409:1446 The predominant oligosaccharide chains 1409:1446 The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. 8670172 15 47 theme phospholipase 2124:2136 arg1 D 2138:2138 a mammalian phospholipase D 2112:2138 a mammalian phospholipase D 2112:2138 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 11 48 theme anion-exchange 1538:1551 arg1 chromatography 1553:1566 Mono Q anion-exchange chromatography 1531:1566 Mono Q anion-exchange chromatography 1531:1566 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 2 49 theme lipid-free 419:428 arg1 forms 430:434 soluble lipid-free forms 411:434 soluble lipid-free forms of CD59 411:442 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 5 50 theme isolated 823:830 arg1 T9 852:853 T9 852:853 T9 852:853 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 50 theme isolated 823:830 arg1 peptide 843:849 the isolated C-terminal peptide 819:849 the isolated C-terminal peptide (T9) 819:854 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 1 51 theme complex 296:302 arg1 CD59 205:208 CD59 205:208 CD59 (protectin) 205:220 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 1 51 theme complex 296:302 arg1 inhibitor 263:271 a glycophosphoinositol (GPI)-anchored inhibitor 225:271 a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells 225:370 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 9 52 link N-linked 1312:1319 arg1 chain 1339:1343 The N-linked carbohydrate side chain 1308:1343 The N-linked carbohydrate side chain of CD59u (at Asn-18) 1308:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 2 53 theme body 465:468 arg1 fluids 470:475 human body fluids 459:475 human body fluids 459:475 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 3 54 theme exoglycosidase 656:669 arg1 digestion 671:679 sequential exoglycosidase digestion 645:679 sequential exoglycosidase digestion 645:679 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 11 55 theme Mono 1531:1534 arg1 chromatography 1553:1566 Mono Q anion-exchange chromatography 1531:1566 Mono Q anion-exchange chromatography 1531:1566 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 1 56 theme complement 307:316 arg1 complex 296:302 the membrane attack complex 276:302 the membrane attack complex of complement 276:316 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 9 57 theme considerable 1381:1392 arg1 heterogeneity 1394:1406 considerable heterogeneity 1381:1406 considerable heterogeneity 1381:1406 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 5 58 theme Mass 802:805 arg1 analysis 807:814 Mass analysis 802:814 Mass analysis of the isolated C-terminal peptide (T9) 802:854 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 14 59 theme GPI 1883:1885 arg1 anchor 1887:1892 the GPI anchor 1879:1892 the GPI anchor 1879:1892 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 3 60 theme laser-desorption 701:716 arg1 MALDI-MS 737:744 MALDI-MS 737:744 MALDI-MS 737:744 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 60 theme laser-desorption 701:716 arg1 spectrometry 723:734 matrix-assisted laser-desorption mass spectrometry 685:734 matrix-assisted laser-desorption mass spectrometry (MALDI-MS) 685:745 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 61 theme detailed 503:510 arg1 composition 523:533 the detailed structural composition 499:533 the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) 499:589 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 1 62 theme -anchored 253:261 arg1 CD59 205:208 CD59 205:208 CD59 (protectin) 205:220 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 1 62 theme -anchored 253:261 arg1 inhibitor 263:271 a glycophosphoinositol (GPI)-anchored inhibitor 225:271 a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells 225:370 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 5 63 from CD59u 963:967 arg1 present 944:950 present 944:950 present 944:950 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 63 from CD59u 963:967 arg1 GPI-anchor 873:882 a GPI-anchor 871:882 a GPI-anchor (at Asn-77) without an inositol-associated phospholipid 871:938 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 2 64 located present 448:454 arg2 forms 430:434 soluble lipid-free forms 411:434 soluble lipid-free forms of CD59 411:442 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 2 64 located present 448:454 arg1 fluids 470:475 human body fluids 459:475 human body fluids 459:475 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 10 65 theme predominant 1413:1423 arg1 chains 1441:1446 The predominant oligosaccharide chains 1409:1446 The predominant oligosaccharide chains 1409:1446 The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. 8670172 0 66 theme soluble 58:64 arg1 CD59 66:69 soluble CD59 58:69 soluble CD59 58:69 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 11 67 theme complement 1660:1669 arg1 complexes 1678:1686 the terminal complement SC5b-8 complexes 1647:1686 the terminal complement SC5b-8 complexes 1647:1686 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 3 68 theme urinary 570:576 arg1 CD59u 584:588 CD59u 584:588 CD59u 584:588 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 68 theme urinary 570:576 arg1 CD59 578:581 the naturally occurring soluble urinary CD59 538:581 the naturally occurring soluble urinary CD59 (CD59u) 538:589 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 13 69 theme erythrocyte 1801:1811 arg1 CD59 1813:1816 erythrocyte CD59 1801:1816 erythrocyte CD59 1801:1816 200-fold lower efficiency than erythrocyte CD59. 8670172 3 70 theme occurring 552:560 arg1 CD59u 584:588 CD59u 584:588 CD59u 584:588 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 70 theme occurring 552:560 arg1 CD59 578:581 the naturally occurring soluble urinary CD59 538:581 the naturally occurring soluble urinary CD59 (CD59u) 538:589 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 5 71 theme C-terminal 832:841 arg1 T9 852:853 T9 852:853 T9 852:853 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 71 theme C-terminal 832:841 arg1 peptide 843:849 the isolated C-terminal peptide 819:849 the isolated C-terminal peptide (T9) 819:854 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 8 72 theme GPI-anchor 1287:1296 arg1 variants 1298:1305 two GPI-anchor variants 1283:1305 two GPI-anchor variants 1283:1305 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 9 73 from chain 1339:1343 arg1 Asn-18 1358:1363 Asn-18 1358:1363 Asn-18 1358:1363 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 5 74 located present 944:950 arg1 CD59u 963:967 soluble CD59u 955:967 soluble CD59u 955:967 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 74 located present 944:950 arg2 present 944:950 present 944:950 present 944:950 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 74 located present 944:950 arg2 GPI-anchor 873:882 a GPI-anchor 871:882 a GPI-anchor (at Asn-77) without an inositol-associated phospholipid 871:938 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 0 75 theme laser-desorption 159:174 arg1 analysis 195:202 laser-desorption mass spectrometric analysis 159:202 laser-desorption mass spectrometric analysis 159:202 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 8 76 theme N-acetylhexosamine-galactose 1237:1264 arg1 arm 1266:1268 an N-acetylhexosamine-galactose arm 1234:1268 an N-acetylhexosamine-galactose arm 1234:1268 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 8 77 attach linked 1224:1229 arg1 arm 1266:1268 an N-acetylhexosamine-galactose arm 1234:1268 an N-acetylhexosamine-galactose arm 1234:1268 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 8 77 attach linked 1224:1229 arg2 acid 1219:1222 Sialic acid 1212:1222 Sialic acid linked to an N-acetylhexosamine-galactose arm 1212:1268 Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. 8670172 14 78 link N-linked 1898:1905 arg1 oligosaccharide 1907:1921 N-linked oligosaccharide 1898:1921 N-linked oligosaccharide 1898:1921 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 11 79 theme urinary 1577:1583 arg1 CD59 1585:1588 urinary CD59 1577:1588 urinary CD59 1577:1588 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 0 80 theme spectrometric 181:193 arg1 analysis 195:202 laser-desorption mass spectrometric analysis 159:202 laser-desorption mass spectrometric analysis 159:202 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 6 81 theme MALDI-MS 1039:1046 arg1 structures 1048:1057 MALDI-MS structures 1039:1057 MALDI-MS structures 1039:1057 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 3 82 theme peptide 597:603 arg1 mapping 605:611 peptide mapping 597:611 peptide mapping 597:611 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 14 83 theme functional 1997:2006 arg1 activity 2008:2015 the full functional activity 1988:2015 the full functional activity of CD59 1988:2023 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 12 84 theme complement 1739:1748 arg1 lysis 1750:1754 complement lysis 1739:1754 complement lysis 1739:1754 Despite binding to C5b-8, soluble CD59u inhibited complement lysis at an approx. 8670172 11 85 theme different 1600:1608 arg1 fractions 1610:1618 nine different fractions 1595:1618 nine different fractions that bound equally well to the terminal complement SC5b-8 complexes 1595:1686 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 9 86 theme carbohydrate 1321:1332 arg1 chain 1339:1343 The N-linked carbohydrate side chain 1308:1343 The N-linked carbohydrate side chain of CD59u (at Asn-18) 1308:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 10 87 theme variable 1509:1516 arg1 sialylation 1518:1528 variable sialylation 1509:1528 variable sialylation 1509:1528 The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. 8670172 5 88 from Asn-77 888:893 arg1 present 944:950 present 944:950 present 944:950 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 88 from Asn-77 888:893 arg1 GPI-anchor 873:882 a GPI-anchor 871:882 a GPI-anchor (at Asn-77) without an inositol-associated phospholipid 871:938 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 5 89 theme soluble 955:961 arg1 CD59u 963:967 soluble CD59u 955:967 soluble CD59u 955:967 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 6 90 theme residue-specific 979:994 arg1 exoglycosidases 996:1010 residue-specific exoglycosidases 979:1010 residue-specific exoglycosidases 979:1010 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 15 91 theme GPI-anchored 2183:2194 arg1 proteins 2196:2203 GPI-anchored proteins 2183:2203 GPI-anchored proteins 2183:2203 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 9 92 theme CD59u 1348:1352 arg1 chain 1339:1343 The N-linked carbohydrate side chain 1308:1343 The N-linked carbohydrate side chain of CD59u (at Asn-18) 1308:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 14 93 theme full 1992:1995 arg1 activity 2008:2015 the full functional activity 1988:2015 the full functional activity of CD59 1988:2023 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 15 94 theme cleavage 2038:2045 arg1 cleavage 2038:2045 cleavage 2038:2045 cleavage 2038:2045 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 15 94 theme cleavage 2038:2045 arg1 site 2030:2033 The site 2026:2033 The site of cleavage between the diradylglycerol phosphate and inositol 2026:2096 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 7 95 theme monosaccharide 1190:1203 arg1 units 1205:1209 one or more monosaccharide units 1178:1209 one or more monosaccharide units 1178:1209 Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. 8670172 9 96 from Asn-18 1358:1363 arg1 CD59u 1348:1352 CD59u 1348:1352 CD59u (at Asn-18) 1348:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 9 96 from Asn-18 1358:1363 arg1 chain 1339:1343 The N-linked carbohydrate side chain 1308:1343 The N-linked carbohydrate side chain of CD59u (at Asn-18) 1308:1364 The N-linked carbohydrate side chain of CD59u (at Asn-18) also displayed considerable heterogeneity. 8670172 4 97 from M 775:775 arg1 MALDI-MS 792:799 MALDI-MS 792:799 MALDI-MS 792:799 CD59u exhibited an average M(r) of 12444 in MALDI-MS. 8670172 1 98 located found 318:322 arg1 endothelia 340:349 endothelia 340:349 endothelia 340:349 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 1 98 located found 318:322 arg1 cells 366:370 epithelial cells 355:370 epithelial cells 355:370 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 1 98 located found 318:322 arg1 cells 333:337 blood cells 327:337 blood cells 327:337 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 1 98 located found 318:322 arg2 inhibitor 263:271 a glycophosphoinositol (GPI)-anchored inhibitor 225:271 a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells 225:370 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 1 98 located found 318:322 arg2 CD59 205:208 CD59 205:208 CD59 (protectin) 205:220 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 6 99 theme variants 1089:1096 arg1 modification 1022:1033 chemical modification 1013:1033 chemical modification 1013:1033 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 6 99 theme variants 1089:1096 arg1 structures 1048:1057 MALDI-MS structures 1039:1057 MALDI-MS structures 1039:1057 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 2 100 theme CD59 439:442 arg1 forms 430:434 soluble lipid-free forms 411:434 soluble lipid-free forms of CD59 411:442 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 11 101 theme Q 1536:1536 arg1 chromatography 1553:1566 Mono Q anion-exchange chromatography 1531:1566 Mono Q anion-exchange chromatography 1531:1566 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 0 102 theme functional 27:36 arg1 characterization 38:53 functional characterization 27:53 functional characterization 27:53 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 1 103 theme attack 289:294 arg1 complex 296:302 the membrane attack complex 276:302 the membrane attack complex of complement 276:316 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 12 104 theme soluble 1715:1721 arg1 CD59u 1723:1727 soluble CD59u 1715:1727 soluble CD59u 1715:1727 Despite binding to C5b-8, soluble CD59u inhibited complement lysis at an approx. 8670172 6 105 theme different 1068:1076 arg1 variants 1089:1096 seven different GPI-anchor variants 1062:1096 seven different GPI-anchor variants 1062:1096 By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. 8670172 3 106 theme sequential 645:654 arg1 digestion 671:679 sequential exoglycosidase digestion 645:679 sequential exoglycosidase digestion 645:679 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 2 107 theme soluble 411:417 arg1 forms 430:434 soluble lipid-free forms 411:434 soluble lipid-free forms of CD59 411:442 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 14 108 theme N-linked 1898:1905 arg1 oligosaccharide 1907:1921 N-linked oligosaccharide 1898:1921 N-linked oligosaccharide 1898:1921 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 2 109 theme human 459:463 arg1 fluids 470:475 human body fluids 459:475 human body fluids 459:475 In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. 8670172 15 110 theme mammalian 2114:2122 arg1 D 2138:2138 a mammalian phospholipase D 2112:2138 a mammalian phospholipase D 2112:2138 The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins. 8670172 0 111 theme CD59 66:69 arg1 composition 11:21 Structural composition 0:21 Structural composition 0:21 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 0 111 theme CD59 66:69 arg1 characterization 38:53 functional characterization 27:53 functional characterization 27:53 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 14 112 theme anchor 1887:1892 arg1 heterogeneity 1857:1869 the structural heterogeneity 1842:1869 the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 1842:1929 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 8670172 3 113 theme matrix-assisted 685:699 arg1 MALDI-MS 737:744 MALDI-MS 737:744 MALDI-MS 737:744 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 3 113 theme matrix-assisted 685:699 arg1 spectrometry 723:734 matrix-assisted laser-desorption mass spectrometry 685:734 matrix-assisted laser-desorption mass spectrometry (MALDI-MS) 685:745 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 11 114 theme SC5b-8 1671:1676 arg1 complexes 1678:1686 the terminal complement SC5b-8 complexes 1647:1686 the terminal complement SC5b-8 complexes 1647:1686 Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b-8 complexes. 8670172 5 115 from present 944:950 arg1 CD59u 963:967 soluble CD59u 955:967 soluble CD59u 955:967 Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59u. 8670172 1 116 theme blood 327:331 arg1 cells 333:337 blood cells 327:337 blood cells 327:337 CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. 8670172 0 117 dep composition 11:21 arg1 heterogeneity 72:84 heterogeneity 72:84 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 0:203 Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis. 8670172 3 118 theme structural 512:521 arg1 composition 523:533 the detailed structural composition 499:533 the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) 499:589 We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59u) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). 8670172 14 119 theme structural 1846:1855 arg1 heterogeneity 1857:1869 the structural heterogeneity 1842:1869 the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 1842:1929 These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. 17558413 6 0 theme tyrosylprotein 949:962 arg1 TPSTs 983:987 TPSTs 983:987 TPSTs 983:987 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 0 theme tyrosylprotein 949:962 arg1 sulfotransferases 964:980 human tyrosylprotein sulfotransferases 943:980 purified human tyrosylprotein sulfotransferases (TPSTs) 934:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 1 theme several 905:911 arg1 peptides 913:920 several peptides 905:920 several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs) 905:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 2 theme tyrosine 1024:1031 arg1 sites 1043:1047 the tyrosine sulfation sites 1020:1047 the tyrosine sulfation sites in mouse lumican and human vitronectin 1020:1086 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 3 theme human 943:947 arg1 TPSTs 983:987 TPSTs 983:987 TPSTs 983:987 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 3 theme human 943:947 arg1 sulfotransferases 964:980 human tyrosylprotein sulfotransferases 943:980 purified human tyrosylprotein sulfotransferases (TPSTs) 934:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 3 4 theme one-step 389:396 arg1 reaction 398:405 a one-step reaction 387:405 a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0 387:486 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 0 5 from Determination 0:12 arg1 proteins 67:74 proteins 67:74 proteins 67:74 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 17558413 0 5 from Determination 0:12 arg1 peptides 54:61 peptides 54:61 peptides 54:61 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 17558413 5 6 used used 752:755 arg2 we 749:750 we 749:750 we 749:750 Since phosphorylation and sulfation of tyrosine are isobaric, we used alkaline phosphatase treatment to distinguish these two modifications. 17558413 6 7 theme sulfation 1033:1041 arg1 sites 1043:1047 the tyrosine sulfation sites 1020:1047 the tyrosine sulfation sites in mouse lumican and human vitronectin 1020:1086 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 3 8 from groups 311:316 arg1 tyrosines 332:340 unsulfated tyrosines 321:340 unsulfated tyrosines 321:340 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 5 9 theme alkaline 757:764 arg1 phosphatase 766:776 alkaline phosphatase 757:776 alkaline phosphatase treatment 757:786 Since phosphorylation and sulfation of tyrosine are isobaric, we used alkaline phosphatase treatment to distinguish these two modifications. 17558413 1 10 theme extracellular 186:198 arg1 space 200:204 extracellular space 186:204 extracellular space 186:204 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 5 11 theme phosphatase 766:776 arg1 treatment 778:786 alkaline phosphatase treatment 757:786 alkaline phosphatase treatment 757:786 Since phosphorylation and sulfation of tyrosine are isobaric, we used alkaline phosphatase treatment to distinguish these two modifications. 17558413 6 12 theme peptides 913:920 arg1 sulfation 892:900 sulfation 892:900 sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs) 892:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 2 13 from sites 259:263 arg1 proteins 292:299 proteins 292:299 proteins 292:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 4 14 theme sulfotyrosine 505:517 arg1 presence 493:500 The presence 489:500 The presence of sulfotyrosine 489:517 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 1 15 theme Tyrosine 77:84 arg1 modification 126:137 a key post-translational modification 101:137 a key post-translational modification that regulates protein-protein interactions in extracellular space 101:204 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 1 15 theme Tyrosine 77:84 arg1 O-sulfation 86:96 Tyrosine O-sulfation 77:96 Tyrosine O-sulfation 77:96 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 4 16 theme mass 579:582 arg1 MS/MS 598:602 MS/MS 598:602 MS/MS 598:602 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 4 16 theme mass 579:582 arg1 spectrometry 584:595 tandem mass spectrometry 572:595 tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile 572:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 6 17 from sites 1043:1047 arg1 lumican 1058:1064 mouse lumican 1052:1064 mouse lumican 1052:1064 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 17 from sites 1043:1047 arg1 vitronectin 1076:1086 human vitronectin 1070:1086 human vitronectin 1070:1086 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 0 18 theme sites 21:25 arg1 Determination 0:12 Determination 0:12 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 0:75 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 17558413 4 19 theme tandem 572:577 arg1 MS/MS 598:602 MS/MS 598:602 MS/MS 598:602 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 4 19 theme tandem 572:577 arg1 spectrometry 584:595 tandem mass spectrometry 572:595 tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile 572:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 3 20 theme Hydroxyl 302:309 arg1 groups 311:316 Hydroxyl groups 302:316 Hydroxyl groups on unsulfated tyrosines 302:340 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 3 21 attach presence 456:463 arg1 pH 481:482 pH 7.0 481:486 pH 7.0 481:486 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 3 21 attach presence 456:463 arg2 imidazole 468:476 imidazole 468:476 imidazole 468:476 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 3 22 theme sulfosuccinimidyl 413:429 arg1 acetate 431:437 sulfosuccinimidyl acetate 413:437 sulfosuccinimidyl acetate (S-NHSAc) 413:447 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 3 22 theme sulfosuccinimidyl 413:429 arg1 S-NHSAc 440:446 S-NHSAc 440:446 S-NHSAc 440:446 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 6 23 dep identified 854:863 arg1 Using 828:832 Using 828:832 Using this methodology 828:849 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 4 24 theme sulfuryl 644:651 arg1 labile 679:684 labile 679:684 labile 679:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 4 24 theme sulfuryl 644:651 arg1 sulfotyrosine 662:674 sulfotyrosine 662:674 sulfotyrosine 662:674 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 4 24 theme sulfuryl 644:651 arg1 group 653:657 the sulfuryl group 640:657 the sulfuryl group of sulfotyrosine 640:674 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 0 25 theme tyrosine 30:37 arg1 O-sulfation 39:49 tyrosine O-sulfation 30:49 tyrosine O-sulfation 30:49 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 17558413 4 26 theme spectrometry 584:595 arg1 analysis 605:612 tandem mass spectrometry (MS/MS) analysis 572:612 tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile 572:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 6 27 theme sulfation 892:900 arg1 sulfation 892:900 sulfation 892:900 sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs) 892:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 27 theme sulfation 892:900 arg1 sites 869:873 the sites 865:873 the sites 865:873 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 27 theme sulfation 892:900 arg1 order 883:887 the order 879:887 the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs) 879:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 2 28 from O-sulfation 277:287 arg1 proteins 292:299 proteins 292:299 proteins 292:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 4 29 theme sulfotyrosine 662:674 arg1 labile 679:684 labile 679:684 labile 679:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 4 29 theme sulfotyrosine 662:674 arg1 sulfotyrosine 662:674 sulfotyrosine 662:674 sulfotyrosine 662:674 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 4 29 theme sulfotyrosine 662:674 arg1 group 653:657 the sulfuryl group 640:657 the sulfuryl group of sulfotyrosine 640:674 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 3 30 theme imidazole 468:476 arg1 presence 456:463 the presence 452:463 the presence of imidazole at pH 7.0 452:486 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 4 31 theme free 552:555 arg1 tyrosine 557:564 free tyrosine 552:564 free tyrosine 552:564 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 3 32 from pH 481:482 arg1 presence 456:463 the presence 452:463 the presence of imidazole at pH 7.0 452:486 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 1 33 theme key 103:105 arg1 modification 126:137 a key post-translational modification 101:137 a key post-translational modification that regulates protein-protein interactions in extracellular space 101:204 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 1 33 theme key 103:105 arg1 O-sulfation 86:96 Tyrosine O-sulfation 77:96 Tyrosine O-sulfation 77:96 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 0 34 theme O-sulfation 39:49 arg1 sites 21:25 the sites 17:25 the sites of tyrosine O-sulfation 17:49 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 17558413 0 34 theme O-sulfation 39:49 arg1 O-sulfation 39:49 tyrosine O-sulfation 30:49 tyrosine O-sulfation 30:49 Determination of the sites of tyrosine O-sulfation in peptides and proteins. 17558413 5 35 theme tyrosine 726:733 arg1 sulfation 713:721 sulfation 713:721 sulfation 713:721 Since phosphorylation and sulfation of tyrosine are isobaric, we used alkaline phosphatase treatment to distinguish these two modifications. 17558413 5 35 theme tyrosine 726:733 arg1 phosphorylation 693:707 phosphorylation 693:707 phosphorylation 693:707 Since phosphorylation and sulfation of tyrosine are isobaric, we used alkaline phosphatase treatment to distinguish these two modifications. 17558413 1 36 theme post-translational 107:124 arg1 modification 126:137 a key post-translational modification 101:137 a key post-translational modification that regulates protein-protein interactions in extracellular space 101:204 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 1 36 theme post-translational 107:124 arg1 O-sulfation 86:96 Tyrosine O-sulfation 77:96 Tyrosine O-sulfation 77:96 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 4 37 theme tyrosine 557:564 arg1 detection 539:547 the detection 535:547 the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile 535:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 2 38 theme subtractive 221:231 arg1 strategy 233:240 a subtractive strategy 219:240 a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins 219:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 2 39 from proteins 292:299 arg1 sites 259:263 the sites 255:263 the sites of tyrosine O-sulfation in proteins 255:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 2 39 from proteins 292:299 arg1 O-sulfation 277:287 tyrosine O-sulfation 268:287 tyrosine O-sulfation in proteins 268:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 6 40 theme human 1070:1074 arg1 vitronectin 1076:1086 human vitronectin 1070:1086 human vitronectin 1070:1086 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 3 41 theme unsulfated 321:330 arg1 tyrosines 332:340 unsulfated tyrosines 321:340 unsulfated tyrosines 321:340 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 2 42 theme O-sulfation 277:287 arg1 sites 259:263 the sites 255:263 the sites of tyrosine O-sulfation in proteins 255:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 2 42 theme O-sulfation 277:287 arg1 O-sulfation 277:287 tyrosine O-sulfation 268:287 tyrosine O-sulfation in proteins 268:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 4 43 from labile 679:684 arg1 conditions 620:629 conditions 620:629 conditions in which the sulfuryl group of sulfotyrosine is labile 620:684 The presence of sulfotyrosine is indicated by the detection of free tyrosine after tandem mass spectrometry (MS/MS) analysis under conditions in which the sulfuryl group of sulfotyrosine is labile. 17558413 3 44 theme stoichiometric 357:370 arg1 acetylation 372:382 stoichiometric acetylation 357:382 stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0 357:486 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 3 45 from acetylation 372:382 arg1 reaction 398:405 a one-step reaction 387:405 a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0 387:486 Hydroxyl groups on unsulfated tyrosines are blocked by stoichiometric acetylation in a one-step reaction using sulfosuccinimidyl acetate (S-NHSAc) in the presence of imidazole at pH 7.0. 17558413 2 46 theme tyrosine 268:275 arg1 O-sulfation 277:287 tyrosine O-sulfation 268:287 tyrosine O-sulfation in proteins 268:299 We describe a subtractive strategy to determine the sites of tyrosine O-sulfation in proteins. 17558413 6 47 theme purified 934:941 arg1 TPSTs 983:987 TPSTs 983:987 TPSTs 983:987 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 6 47 theme purified 934:941 arg1 sulfotransferases 964:980 human tyrosylprotein sulfotransferases 943:980 purified human tyrosylprotein sulfotransferases (TPSTs) 934:988 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 17558413 1 48 theme protein-protein 154:168 arg1 interactions 170:181 protein-protein interactions 154:181 protein-protein interactions 154:181 Tyrosine O-sulfation is a key post-translational modification that regulates protein-protein interactions in extracellular space. 17558413 6 49 theme mouse 1052:1056 arg1 lumican 1058:1064 mouse lumican 1052:1064 mouse lumican 1052:1064 Using this methodology we identified the sites and the order of sulfation of several peptides mediated by purified human tyrosylprotein sulfotransferases (TPSTs), and unambiguously determined the tyrosine sulfation sites in mouse lumican and human vitronectin. 12889478 2 0 theme recognition 563:573 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 0 theme recognition 563:573 arg1 determinant 575:585 a recognition determinant 561:585 a recognition determinant within their heavily glycosylated mucin domains 561:633 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 5 1 link O-linked 974:981 arg1 structures 988:997 O-linked sLex structures 974:997 O-linked sLex structures 974:997 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 2 2 theme mucin 621:625 arg1 domains 627:633 their heavily glycosylated mucin domains 594:633 their heavily glycosylated mucin domains 594:633 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 7 3 dep cells 1402:1406 arg1 addition 1378:1385 addition 1378:1385 addition 1378:1385 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 5 4 theme tyrosine 952:959 arg1 residues 961:968 two tyrosine residues 948:968 two tyrosine residues 948:968 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 0 5 theme L-selectin 57:66 arg1 ligand 68:73 an L-selectin ligand 54:73 an L-selectin ligand 54:73 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 0 5 theme L-selectin 57:66 arg1 Endoglycan 0:9 Endoglycan 0:9 Endoglycan 0:9 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 2 6 theme glycosylated 608:619 arg1 domains 627:633 their heavily glycosylated mucin domains 594:633 their heavily glycosylated mucin domains 594:633 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 1 7 from ligands 392:398 arg1 endothelium 407:417 the endothelium 403:417 the endothelium 403:417 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 7 8 theme endoglycan 1300:1309 arg1 distribution 1284:1295 the distribution 1280:1295 the distribution of endoglycan 1280:1309 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 2 9 theme L-selectin 425:434 arg1 CD34 452:455 CD34 452:455 CD34 452:455 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 9 theme L-selectin 425:434 arg1 podocalyxin 461:471 podocalyxin 461:471 podocalyxin 461:471 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 9 theme L-selectin 425:434 arg1 ligands 436:442 Most L-selectin ligands 420:442 Most L-selectin ligands such as CD34 and podocalyxin 420:471 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 5 10 theme O-linked 974:981 arg1 structures 988:997 O-linked sLex structures 974:997 O-linked sLex structures 974:997 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 2 11 dep structures 503:512 arg1 x 539:539 6-sulfated sialyl Lewis x 515:539 6-sulfated sialyl Lewis x 515:539 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 11 dep structures 503:512 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 11 dep structures 503:512 arg1 6-sulfo-sLex 544:555 6-sulfo-sLex 544:555 6-sulfo-sLex 544:555 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 12 theme Lewis 533:537 arg1 x 539:539 6-sulfated sialyl Lewis x 515:539 6-sulfated sialyl Lewis x 515:539 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 12 theme Lewis 533:537 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 1 13 theme several 362:368 arg1 ligands 392:398 several carbohydrate-modified ligands 362:398 several carbohydrate-modified ligands on the endothelium 362:417 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 2 14 theme carbohydrate 490:501 arg1 x 539:539 6-sulfated sialyl Lewis x 515:539 6-sulfated sialyl Lewis x 515:539 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 14 theme carbohydrate 490:501 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 14 theme carbohydrate 490:501 arg1 determinant 575:585 a recognition determinant 561:585 a recognition determinant within their heavily glycosylated mucin domains 561:633 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 14 theme carbohydrate 490:501 arg1 6-sulfo-sLex 544:555 6-sulfo-sLex 544:555 6-sulfo-sLex 544:555 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 0 15 with modification 83:94 arg1 sulfation 110:118 tyrosine sulfation 101:118 tyrosine sulfation 101:118 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 0 15 with modification 83:94 arg1 x 137:137 sialyl Lewis x 124:137 sialyl Lewis x 124:137 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 4 16 theme family 773:778 arg1 members 757:763 the two other members 743:763 the two other members of this family 743:778 We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. 12889478 1 17 theme lymphocyte 147:156 arg1 homing 158:163 lymphocyte homing 147:163 lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking 147:234 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 1 18 theme carbohydrate-modified 370:390 arg1 ligands 392:398 several carbohydrate-modified ligands 362:398 several carbohydrate-modified ligands on the endothelium 362:417 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 0 19 theme sialyl 124:129 arg1 x 137:137 sialyl Lewis x 124:137 sialyl Lewis x 124:137 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 1 20 theme leukocytes 252:261 arg1 rolling 241:247 the rolling 237:247 the rolling of leukocytes on vascular endothelium 237:285 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 5 21 theme amino-terminal 1043:1056 arg1 region 1058:1063 its highly acidic amino-terminal region 1025:1063 its highly acidic amino-terminal region 1025:1063 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 7 22 theme endothelial 1390:1400 arg1 cells 1402:1406 endothelial cells 1390:1406 endothelial cells 1390:1406 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 1 23 from leukocytes 347:356 arg1 endothelium 407:417 the endothelium 403:417 the endothelium 403:417 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 1 24 from rolling 241:247 arg1 endothelium 275:285 vascular endothelium 266:285 vascular endothelium 266:285 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 5 25 theme acidic 1036:1041 arg1 region 1058:1063 its highly acidic amino-terminal region 1025:1063 its highly acidic amino-terminal region 1025:1063 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 4 26 dep endoglycan 726:735 arg1 podocalyxin 790:800 podocalyxin 790:800 podocalyxin 790:800 We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. 12889478 4 26 dep endoglycan 726:735 arg1 CD34 781:784 CD34 781:784 CD34 781:784 We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. 12889478 1 27 theme secondary 168:176 arg1 organs 187:192 secondary lymphoid organs 168:192 secondary lymphoid organs 168:192 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 1 28 theme vascular 266:273 arg1 endothelium 275:285 vascular endothelium 266:285 vascular endothelium 266:285 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 3 29 theme new 675:677 arg1 member 679:684 a new member 673:684 a new member of the CD34 family 673:703 We recently identified endoglycan as a new member of the CD34 family. 12889478 3 29 theme new 675:677 arg1 endoglycan 659:668 endoglycan 659:668 endoglycan 659:668 We recently identified endoglycan as a new member of the CD34 family. 12889478 4 30 theme other 751:755 arg1 members 757:763 the two other members 743:763 the two other members of this family 743:778 We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. 12889478 1 31 theme lymphoid 178:185 arg1 organs 187:192 secondary lymphoid organs 168:192 secondary lymphoid organs 168:192 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 1 32 from L-selectin 333:342 arg1 ligands 392:398 several carbohydrate-modified ligands 362:398 several carbohydrate-modified ligands on the endothelium 362:417 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 1 32 from L-selectin 333:342 arg1 leukocytes 347:356 leukocytes 347:356 leukocytes 347:356 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 2 33 theme Most 420:423 arg1 CD34 452:455 CD34 452:455 CD34 452:455 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 33 theme Most 420:423 arg1 podocalyxin 461:471 podocalyxin 461:471 podocalyxin 461:471 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 33 theme Most 420:423 arg1 ligands 436:442 Most L-selectin ligands 420:442 Most L-selectin ligands such as CD34 and podocalyxin 420:471 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 34 gly glycosylated 608:619 arg1 domains 627:633 their heavily glycosylated mucin domains 594:633 their heavily glycosylated mucin domains 594:633 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 35 theme sulfated 481:488 arg1 x 539:539 6-sulfated sialyl Lewis x 515:539 6-sulfated sialyl Lewis x 515:539 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 35 theme sulfated 481:488 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 35 theme sulfated 481:488 arg1 determinant 575:585 a recognition determinant 561:585 a recognition determinant within their heavily glycosylated mucin domains 561:633 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 35 theme sulfated 481:488 arg1 6-sulfo-sLex 544:555 6-sulfo-sLex 544:555 6-sulfo-sLex 544:555 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 6 36 with parallels 1100:1108 arg1 ligand 1135:1140 a leukocyte ligand 1123:1140 a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions 1123:1271 Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. 12889478 6 36 with parallels 1100:1108 arg1 PSGL-1 1115:1120 PSGL-1 1115:1120 PSGL-1 1115:1120 Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. 12889478 7 37 theme endoglycan-mediated 1461:1479 arg1 events 1490:1495 endoglycan-mediated adhesion events 1461:1495 endoglycan-mediated adhesion events 1461:1495 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 0 38 theme family 33:38 arg1 Endoglycan 0:9 Endoglycan 0:9 Endoglycan 0:9 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 0 38 theme family 33:38 arg1 member 14:19 a member 12:19 a member of the CD34 family 12:38 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 0 39 theme Lewis 131:135 arg1 x 137:137 sialyl Lewis x 124:137 sialyl Lewis x 124:137 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 7 40 theme potential 1438:1446 arg1 settings 1448:1455 several potential settings 1430:1455 several potential settings for endoglycan-mediated adhesion events 1430:1495 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 0 41 theme CD34 28:31 arg1 family 33:38 the CD34 family 24:38 the CD34 family 24:38 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 0 42 theme tyrosine 101:108 arg1 sulfation 110:118 tyrosine sulfation 101:118 tyrosine sulfation 101:118 Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. 12889478 3 43 theme CD34 693:696 arg1 family 698:703 the CD34 family 689:703 the CD34 family 689:703 We recently identified endoglycan as a new member of the CD34 family. 12889478 4 44 theme L-selectin 821:830 arg1 endoglycan 726:735 endoglycan 726:735 endoglycan 726:735 We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. 12889478 4 44 theme L-selectin 821:830 arg1 ligand 832:837 a L-selectin ligand 819:837 a L-selectin ligand 819:837 We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. 12889478 7 45 theme leukocyte 1349:1357 arg1 subpopulations 1359:1372 leukocyte subpopulations 1349:1372 leukocyte subpopulations 1349:1372 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 1 46 theme transient 302:310 arg1 interactions 312:323 transient interactions 302:323 transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium 302:417 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 3 47 theme family 698:703 arg1 member 679:684 a new member 673:684 a new member of the CD34 family 673:703 We recently identified endoglycan as a new member of the CD34 family. 12889478 3 47 theme family 698:703 arg1 endoglycan 659:668 endoglycan 659:668 endoglycan 659:668 We recently identified endoglycan as a new member of the CD34 family. 12889478 6 48 theme platelet-leukocyte 1241:1258 arg1 interactions 1260:1271 leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions 1193:1271 interactions 1260:1271 Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. 12889478 7 49 theme hematopoietic 1320:1332 arg1 precursors 1334:1343 hematopoietic precursors 1320:1343 hematopoietic precursors 1320:1343 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 7 50 theme adhesion 1481:1488 arg1 events 1490:1495 endoglycan-mediated adhesion events 1461:1495 endoglycan-mediated adhesion events 1461:1495 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 5 51 theme sLex 983:986 arg1 structures 988:997 O-linked sLex structures 974:997 O-linked sLex structures 974:997 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 7 52 theme several 1430:1436 arg1 settings 1448:1455 several potential settings 1430:1455 several potential settings for endoglycan-mediated adhesion events 1430:1495 Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan-mediated adhesion events. 12889478 6 53 theme leukocyte 1125:1133 arg1 ligand 1135:1140 a leukocyte ligand 1123:1140 a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions 1123:1271 Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. 12889478 6 53 theme leukocyte 1125:1133 arg1 PSGL-1 1115:1120 PSGL-1 1115:1120 PSGL-1 1115:1120 Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. 12889478 5 54 from sulfation 935:943 arg1 structures 988:997 O-linked sLex structures 974:997 O-linked sLex structures 974:997 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 5 54 from sulfation 935:943 arg1 residues 961:968 two tyrosine residues 948:968 two tyrosine residues 948:968 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 2 55 theme sialyl 526:531 arg1 x 539:539 6-sulfated sialyl Lewis x 515:539 6-sulfated sialyl Lewis x 515:539 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 55 theme sialyl 526:531 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 5 56 theme different 870:878 arg1 mechanism 888:896 a different binding mechanism 868:896 a different binding mechanism 868:896 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 6 57 theme striking 1091:1098 arg1 parallels 1100:1108 striking parallels 1091:1108 striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions 1091:1271 Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. 12889478 1 58 theme inflammatory 211:222 arg1 trafficking 224:234 inflammatory trafficking 211:234 inflammatory trafficking 211:234 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 2 59 theme 6-sulfated 515:524 arg1 x 539:539 6-sulfated sialyl Lewis x 515:539 6-sulfated sialyl Lewis x 515:539 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 2 59 theme 6-sulfated 515:524 arg1 structures 503:512 sulfated carbohydrate structures 481:512 sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) 481:556 Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. 12889478 5 60 theme binding 880:886 arg1 mechanism 888:896 a different binding mechanism 868:896 a different binding mechanism 868:896 However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. 12889478 1 61 theme trafficking 224:234 arg1 instances 198:206 instances 198:206 instances 198:206 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 12889478 1 61 theme trafficking 224:234 arg1 organs 187:192 secondary lymphoid organs 168:192 secondary lymphoid organs 168:192 During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. 19528533 4 0 theme protease 877:884 arg1 inhibition 886:895 optimal protease inhibition 869:895 optimal protease inhibition 869:895 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 6 1 theme PZ 1341:1342 arg1 PZ 1341:1342 PZ 1341:1342 PZ 1341:1342 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 1 theme PZ 1341:1342 arg1 domains 1330:1336 the gamma-carboxy-glutamic acid-containing domains 1287:1336 the gamma-carboxy-glutamic acid-containing domains of PZ and FXa 1287:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 1 theme PZ 1341:1342 arg1 FXa 1348:1350 FXa 1348:1350 FXa 1348:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 4 2 theme optimal 869:875 arg1 inhibition 886:895 optimal protease inhibition 869:895 optimal protease inhibition 869:895 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 1 3 theme PZ 364:365 arg1 structure 351:359 A 2.3A resolution crystal structure 325:359 A 2.3A resolution crystal structure of PZ complexed with ZPI 325:384 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 4 4 theme thermal 980:986 arg1 stability 988:996 thermal stability 980:996 thermal stability 980:996 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 4 5 theme inhibition 1036:1045 arg1 efficiency 1013:1022 efficiency 1013:1022 efficiency of protease inhibition 1013:1045 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 6 6 theme FXa 1348:1350 arg1 PZ 1341:1342 PZ 1341:1342 PZ 1341:1342 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 6 theme FXa 1348:1350 arg1 domains 1330:1336 the gamma-carboxy-glutamic acid-containing domains 1287:1336 the gamma-carboxy-glutamic acid-containing domains of PZ and FXa 1287:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 6 theme FXa 1348:1350 arg1 FXa 1348:1350 FXa 1348:1350 FXa 1348:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 7 from surfaces 1397:1404 arg1 membranes 1428:1436 platelet and other membranes 1409:1436 membranes 1428:1436 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 3 8 theme molecule 771:778 arg1 core 759:762 the hydrophobic core 743:762 the hydrophobic core of the molecule 743:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 3 9 theme charged 713:719 arg1 residue 721:727 a negatively charged residue 700:727 a negatively charged residue buried within the hydrophobic core of the molecule 700:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 5 10 theme exosite 1202:1208 arg1 interactions 1210:1221 exosite interactions 1202:1221 exosite interactions 1202:1221 The structure of ZPI shows negatively and positively charged surfaces on top of the molecule, in keeping with mutagenesis studies in this work indicating exosite interactions with FXa when it docks on top of ZPI. 19528533 1 11 theme typical 406:412 arg1 ZPI 397:399 ZPI 397:399 ZPI 397:399 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 11 theme typical 406:412 arg1 inhibitor 430:438 a typical serine protease inhibitor 404:438 a typical serine protease inhibitor 404:438 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 3 12 contain has 665:667 arg2 region 688:693 an unusual shutter region 669:693 an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule 669:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 3 12 contain has 665:667 arg1 ZPI 661:663 ZPI 661:663 ZPI 661:663 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 6 13 theme platelet 1409:1416 arg1 membranes 1428:1436 platelet and other membranes 1409:1436 membranes 1428:1436 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 2 14 theme hydrophobic 635:645 arg1 interactions 647:658 extensive ionic and hydrophobic interactions 615:658 extensive ionic and hydrophobic interactions 615:658 The 2 molecules bind with fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions. 19528533 0 15 theme PZ-dependent 167:178 arg1 ZPI 191:193 ZPI 191:193 ZPI 191:193 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 15 theme PZ-dependent 167:178 arg1 inhibitor 180:188 PZ-dependent inhibitor 167:188 PZ-dependent inhibitor (ZPI) 167:194 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 6 16 theme complexed 1495:1503 arg1 ZPI 1505:1507 the complexed ZPI 1491:1507 the complexed ZPI 1491:1507 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 4 17 theme balanced 833:840 arg1 metastability 842:854 the balanced metastability 829:854 the balanced metastability required for optimal protease inhibition 829:895 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 6 18 theme optimal 1444:1450 arg1 proximity 1452:1460 optimal proximity 1444:1460 optimal proximity for the inhibition of FXa by the complexed ZPI 1444:1507 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 4 19 with replacement 936:946 arg1 Asn 953:955 Asn 953:955 Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition 953:1045 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 6 20 theme FXa 1484:1486 arg1 inhibition 1470:1479 the inhibition 1466:1479 the inhibition of FXa by the complexed ZPI 1466:1507 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 5 21 theme mutagenesis 1158:1168 arg1 studies 1170:1176 mutagenesis studies 1158:1176 mutagenesis studies in this work indicating exosite interactions with FXa when it docks on top of ZPI 1158:1258 The structure of ZPI shows negatively and positively charged surfaces on top of the molecule, in keeping with mutagenesis studies in this work indicating exosite interactions with FXa when it docks on top of ZPI. 19528533 1 22 theme serine 458:463 arg1 fold 474:477 a serine protease fold 456:477 a serine protease fold with distorted oxyanion hole and S1 pocket 456:520 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 6 23 theme acid-containing 1314:1328 arg1 PZ 1341:1342 PZ 1341:1342 PZ 1341:1342 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 23 theme acid-containing 1314:1328 arg1 domains 1330:1336 the gamma-carboxy-glutamic acid-containing domains 1287:1336 the gamma-carboxy-glutamic acid-containing domains of PZ and FXa 1287:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 23 theme acid-containing 1314:1328 arg1 FXa 1348:1350 FXa 1348:1350 FXa 1348:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 5 24 from studies 1170:1176 arg1 work 1186:1189 this work 1181:1189 this work indicating exosite interactions with FXa when it docks on top of ZPI 1181:1258 The structure of ZPI shows negatively and positively charged surfaces on top of the molecule, in keeping with mutagenesis studies in this work indicating exosite interactions with FXa when it docks on top of ZPI. 19528533 1 25 theme protease 465:472 arg1 fold 474:477 a serine protease fold 456:477 a serine protease fold with distorted oxyanion hole and S1 pocket 456:520 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 6 26 theme phospholipid 1384:1395 arg1 surfaces 1397:1404 the same phospholipid surfaces 1375:1404 the same phospholipid surfaces on platelet and other membranes 1375:1436 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 0 27 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of protein Z-dependent inhibitor complex 0:57 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 3 28 theme hydrophobic 747:757 arg1 core 759:762 the hydrophobic core 743:762 the hydrophobic core of the molecule 743:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 6 29 theme same 1379:1382 arg1 surfaces 1397:1404 the same phospholipid surfaces 1375:1404 the same phospholipid surfaces on platelet and other membranes 1375:1436 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 0 30 theme protein 21:27 arg1 complex 51:57 protein Z-dependent inhibitor complex 21:57 protein Z-dependent inhibitor complex 21:57 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 5 31 theme ZPI 1065:1067 arg1 structure 1052:1060 The structure 1048:1060 The structure of ZPI 1048:1067 The structure of ZPI shows negatively and positively charged surfaces on top of the molecule, in keeping with mutagenesis studies in this work indicating exosite interactions with FXa when it docks on top of ZPI. 19528533 0 32 theme membrane 110:117 arg1 inhibition 119:128 the membrane inhibition 106:128 the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease 106:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 2 33 theme complementary 555:567 arg1 surfaces 569:576 fully complementary surfaces 549:576 fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions 549:658 The 2 molecules bind with fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions. 19528533 2 34 theme ionic 625:629 arg1 interactions 647:658 extensive ionic and hydrophobic interactions 615:658 extensive ionic and hydrophobic interactions 615:658 The 2 molecules bind with fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions. 19528533 1 35 theme serine 414:419 arg1 ZPI 397:399 ZPI 397:399 ZPI 397:399 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 35 theme serine 414:419 arg1 inhibitor 430:438 a typical serine protease inhibitor 404:438 a typical serine protease inhibitor 404:438 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 36 theme oxyanion 494:501 arg1 hole 503:506 distorted oxyanion hole 484:506 distorted oxyanion hole 484:506 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 37 theme distorted 484:492 arg1 hole 503:506 distorted oxyanion hole 484:506 distorted oxyanion hole 484:506 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 0 38 theme inhibitor 41:49 arg1 complex 51:57 protein Z-dependent inhibitor complex 21:57 protein Z-dependent inhibitor complex 21:57 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 39 theme phospholipids 301:313 arg1 presence 289:296 the presence 285:296 the presence of phospholipids and Ca2+ 285:322 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 40 theme Z-dependent 29:39 arg1 complex 51:57 protein Z-dependent inhibitor complex 21:57 protein Z-dependent inhibitor complex 21:57 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 3 41 theme unusual 672:678 arg1 region 688:693 an unusual shutter region 669:693 an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule 669:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 1 42 contain has 452:454 arg2 fold 474:477 a serine protease fold 456:477 a serine protease fold with distorted oxyanion hole and S1 pocket 456:520 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 42 contain has 452:454 arg1 PZ 449:450 PZ 449:450 PZ 449:450 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 43 with fold 474:477 arg1 hole 503:506 distorted oxyanion hole 484:506 distorted oxyanion hole 484:506 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 43 with fold 474:477 arg1 pocket 515:520 S1 pocket 512:520 S1 pocket 512:520 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 6 44 theme other 1422:1426 arg1 membranes 1428:1436 platelet and other membranes 1409:1436 membranes 1428:1436 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 0 45 theme protease 246:253 arg1 inhibition 216:225 the inhibition 212:225 the inhibition of the coagulation protease 212:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 3 46 theme shutter 680:686 arg1 region 688:693 an unusual shutter region 669:693 an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule 669:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 0 47 theme X. 140:141 arg1 PZ 154:155 PZ 154:155 PZ 154:155 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 47 theme X. 140:141 arg1 Z 151:151 X. Protein Z 140:151 factor X. Protein Z (PZ) 133:156 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 1 48 theme 2.3A 327:330 arg1 structure 351:359 A 2.3A resolution crystal structure 325:359 A 2.3A resolution crystal structure of PZ complexed with ZPI 325:384 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 0 49 theme coagulation 234:244 arg1 protease 246:253 the coagulation protease 230:253 the coagulation protease 230:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 6 50 theme gamma-carboxy-glutamic 1291:1312 arg1 PZ 1341:1342 PZ 1341:1342 PZ 1341:1342 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 50 theme gamma-carboxy-glutamic 1291:1312 arg1 domains 1330:1336 the gamma-carboxy-glutamic acid-containing domains 1287:1336 the gamma-carboxy-glutamic acid-containing domains of PZ and FXa 1287:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 6 50 theme gamma-carboxy-glutamic 1291:1312 arg1 FXa 1348:1350 FXa 1348:1350 FXa 1348:1350 As modeled in this study, the gamma-carboxy-glutamic acid-containing domains of PZ and FXa enable them to bind to the same phospholipid surfaces on platelet and other membranes, with optimal proximity for the inhibition of FXa by the complexed ZPI. 19528533 0 51 theme complex 51:57 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of protein Z-dependent inhibitor complex 0:57 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 52 theme factor 133:138 arg1 inhibition 119:128 the membrane inhibition 106:128 the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease 106:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 1 53 theme resolution 332:341 arg1 structure 351:359 A 2.3A resolution crystal structure 325:359 A 2.3A resolution crystal structure of PZ complexed with ZPI 325:384 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 54 theme crystal 343:349 arg1 structure 351:359 A 2.3A resolution crystal structure 325:359 A 2.3A resolution crystal structure of PZ complexed with ZPI 325:384 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 0 55 theme activated 256:264 arg1 X 273:273 activated factor X 256:273 activated factor X (FXa) 256:279 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 55 theme activated 256:264 arg1 cofactor 94:101 a cofactor 92:101 a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease 92:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 55 theme activated 256:264 arg1 FXa 276:278 FXa 276:278 FXa 276:278 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 56 from cofactor 94:101 arg1 inhibition 119:128 the membrane inhibition 106:128 the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease 106:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 1 57 theme protease 421:428 arg1 ZPI 397:399 ZPI 397:399 ZPI 397:399 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 1 57 theme protease 421:428 arg1 inhibitor 430:438 a typical serine protease inhibitor 404:438 a typical serine protease inhibitor 404:438 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 0 58 theme Ca2+ 319:322 arg1 presence 289:296 the presence 285:296 the presence of phospholipids and Ca2+ 285:322 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 59 theme Protein 143:149 arg1 PZ 154:155 PZ 154:155 PZ 154:155 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 59 theme Protein 143:149 arg1 Z 151:151 X. Protein Z 140:151 factor X. Protein Z (PZ) 133:156 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 3 60 with region 688:693 arg1 residue 721:727 a negatively charged residue 700:727 a negatively charged residue buried within the hydrophobic core of the molecule 700:778 ZPI has an unusual shutter region with a negatively charged residue buried within the hydrophobic core of the molecule. 19528533 0 61 dep factor 133:138 arg1 PZ 154:155 PZ 154:155 PZ 154:155 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 61 dep factor 133:138 arg1 Z 151:151 X. Protein Z 140:151 factor X. Protein Z (PZ) 133:156 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 1 62 theme S1 512:513 arg1 pocket 515:520 S1 pocket 512:520 S1 pocket 512:520 A 2.3A resolution crystal structure of PZ complexed with ZPI shows that ZPI is a typical serine protease inhibitor and that PZ has a serine protease fold with distorted oxyanion hole and S1 pocket. 19528533 4 63 theme protease 1027:1034 arg1 inhibition 1036:1045 protease inhibition 1027:1045 protease inhibition 1027:1045 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 0 64 theme protein 69:75 arg1 Z 77:77 protein Z 69:77 protein Z 69:77 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 64 theme protein 69:75 arg1 cofactor 94:101 a cofactor 92:101 a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease 92:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 65 theme factor 266:271 arg1 X 273:273 activated factor X 256:273 activated factor X (FXa) 256:279 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 65 theme factor 266:271 arg1 cofactor 94:101 a cofactor 92:101 a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease 92:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 65 theme factor 266:271 arg1 FXa 276:278 FXa 276:278 FXa 276:278 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 4 66 theme unique 786:791 arg1 213 797:799 213 797:799 213 797:799 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 4 66 theme unique 786:791 arg1 Asp 793:795 This unique Asp 781:795 This unique Asp(213) 781:800 This unique Asp(213) is critical in maintaining the balanced metastability required for optimal protease inhibition, especially when PZ is bound, with its replacement with Asn resulting in increased thermal stability, but decreased efficiency of protease inhibition. 19528533 2 67 dep surfaces 569:576 arg1 spanning 578:585 spanning 578:585 spanning over 2400A(2) 578:599 The 2 molecules bind with fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions. 19528533 2 67 dep surfaces 569:576 arg1 involving 605:613 involving 605:613 involving extensive ionic and hydrophobic interactions 605:658 The 2 molecules bind with fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions. 19528533 5 68 theme charged 1101:1107 arg1 surfaces 1109:1116 negatively and positively charged surfaces 1075:1116 negatively and positively charged surfaces 1075:1116 The structure of ZPI shows negatively and positively charged surfaces on top of the molecule, in keeping with mutagenesis studies in this work indicating exosite interactions with FXa when it docks on top of ZPI. 19528533 2 69 theme extensive 615:623 arg1 interactions 647:658 extensive ionic and hydrophobic interactions 615:658 extensive ionic and hydrophobic interactions 615:658 The 2 molecules bind with fully complementary surfaces spanning over 2400A(2) and involving extensive ionic and hydrophobic interactions. 19528533 0 70 dep inhibition 119:128 arg1 accelerates 200:210 accelerates 200:210 accelerates the inhibition of the coagulation protease 200:253 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 19528533 0 70 dep inhibition 119:128 arg1 binds 158:162 binds 158:162 binds to PZ-dependent inhibitor (ZPI) 158:194 Crystal structure of protein Z-dependent inhibitor complex shows how protein Z functions as a cofactor in the membrane inhibition of factor X. Protein Z (PZ) binds to PZ-dependent inhibitor (ZPI) and accelerates the inhibition of the coagulation protease, activated factor X (FXa), in the presence of phospholipids and Ca2+. 2579379 6 0 theme % 795:795 arg1 identity 797:804 79% identity 793:804 79% identity to mouse C3 at the nucleotide level 793:840 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 3 1 attach present 404:410 arg2 residues 391:398 four arginine residues 377:398 four arginine residues not present in the mature protein 377:432 The two chains are joined by four arginine residues not present in the mature protein. 2579379 3 1 attach present 404:410 arg1 protein 426:432 the mature protein 415:432 the mature protein 415:432 The two chains are joined by four arginine residues not present in the mature protein. 2579379 4 2 theme protease 610:617 arg1 factor 619:624 the serine protease factor I 599:626 the serine protease factor I 599:626 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 3 theme thiolester 505:514 arg1 site 516:519 the thiolester site 501:519 the thiolester site 501:519 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 3 theme thiolester 505:514 arg1 cleavage 587:594 cleavage 587:594 cleavage by the serine protease factor I 587:626 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 0 4 theme primary 64:70 arg1 structure 72:80 derived primary structure 56:80 derived primary structure 56:80 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 6 5 theme 79 793:794 arg1 % 795:795 % 795:795 % 795:795 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 3 6 from protein 426:432 arg1 present 404:410 present 404:410 present 404:410 The two chains are joined by four arginine residues not present in the mature protein. 2579379 4 7 theme serine 603:608 arg1 factor 619:624 the serine protease factor I 599:626 the serine protease factor I 599:626 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 8 theme Several 435:441 arg1 sites 466:470 Several functionally important sites 435:470 Several functionally important sites 435:470 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 6 9 from level 877:881 arg1 identity 850:857 77% identity 846:857 77% identity at the amino acid level 846:881 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 6 9 from level 877:881 arg1 identity 797:804 79% identity 793:804 79% identity to mouse C3 at the nucleotide level 793:840 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 2 10 theme residues 271:278 arg1 peptide 246:252 a signal peptide 237:252 a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues) 237:345 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 6 11 theme % 848:848 arg1 identity 850:857 77% identity 846:857 77% identity at the amino acid level 846:881 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 2 12 theme acid 266:269 arg1 chain 326:330 the alpha chain 316:330 the alpha chain (992 residues) 316:345 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 12 theme acid 266:269 arg1 residues 271:278 22 amino acid residues 257:278 22 amino acid residues 257:278 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 12 theme acid 266:269 arg1 chain 290:294 the beta chain 281:294 the beta chain (645 residues) 281:309 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 6 13 theme 77 846:847 arg1 % 848:848 % 848:848 % 848:848 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 7 14 theme common 1036:1041 arg1 ancestor 1043:1050 a common ancestor 1034:1050 a common ancestor 1034:1050 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 6 15 theme Human 780:784 arg1 C3 786:787 Human C3 780:787 Human C3 780:787 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 7 16 theme considerable 952:963 arg1 homology 965:972 considerable homology 952:972 considerable homology to C3 952:978 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 2 17 theme amino 260:264 arg1 chain 326:330 the alpha chain 316:330 the alpha chain (992 residues) 316:345 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 17 theme amino 260:264 arg1 residues 271:278 22 amino acid residues 257:278 22 amino acid residues 257:278 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 17 theme amino 260:264 arg1 chain 290:294 the beta chain 281:294 the beta chain (645 residues) 281:309 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 4 18 theme cleavage 587:594 arg1 site 516:519 the thiolester site 501:519 the thiolester site 501:519 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 18 theme cleavage 587:594 arg1 sites 578:582 two sites 574:582 two sites of cleavage by the serine protease factor I 574:626 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 18 theme cleavage 587:594 arg1 site 535:538 the cleavage site 522:538 the cleavage site liberating the anaphylatoxin 522:567 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 18 theme cleavage 587:594 arg1 fragment 650:657 a peptide fragment 640:657 a peptide fragment with leukocyte mobilizing activity 640:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 18 theme cleavage 587:594 arg1 cleavage 587:594 cleavage 587:594 cleavage by the serine protease factor I 587:626 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 1 19 theme human 152:156 arg1 C3 179:180 human complement component C3 152:180 human complement component C3 152:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 3 20 from present 404:410 arg1 protein 426:432 the mature protein 415:432 the mature protein 415:432 The two chains are joined by four arginine residues not present in the mature protein. 2579379 4 21 theme leukocyte 664:672 arg1 activity 685:692 leukocyte mobilizing activity 664:692 leukocyte mobilizing activity 664:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 1 22 theme complement 158:167 arg1 C3 179:180 human complement component C3 152:180 human complement component C3 152:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 0 23 theme complement 6:15 arg1 C3 27:28 Human complement component C3 0:28 Human complement component C3 0:28 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 7 24 dep 2-macroglobulin 903:917 arg1 The 884:886 The 884:886 The 884:886 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 7 25 theme alpha 897:901 arg1 2-macroglobulin 903:917 protease alpha 2-macroglobulin 888:917 protease alpha 2-macroglobulin 888:917 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 1 26 theme component 169:177 arg1 C3 179:180 human complement component C3 152:180 human complement component C3 152:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 0 27 theme Human 0:4 arg1 C3 27:28 Human complement component C3 0:28 Human complement component C3 0:28 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 2 28 theme beta 285:288 arg1 residues 301:308 645 residues 297:308 645 residues 297:308 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 28 theme beta 285:288 arg1 residues 271:278 22 amino acid residues 257:278 22 amino acid residues 257:278 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 28 theme beta 285:288 arg1 chain 290:294 the beta chain 281:294 the beta chain (645 residues) 281:309 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 1 29 theme C3 179:180 arg1 sequence 140:147 derived amino acid sequence 121:147 derived amino acid sequence of human complement component C3 121:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 1 29 theme C3 179:180 arg1 sequence 108:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 5 30 theme attachment 721:730 arg1 one 739:741 one 739:741 one 739:741 At least two carbohydrate attachment sites, one on each chain, have been identified. 2579379 5 30 theme attachment 721:730 arg1 sites 732:736 At least two carbohydrate attachment sites 695:736 At least two carbohydrate attachment sites 695:736 At least two carbohydrate attachment sites, one on each chain, have been identified. 2579379 5 31 theme carbohydrate 708:719 arg1 one 739:741 one 739:741 one 739:741 At least two carbohydrate attachment sites, one on each chain, have been identified. 2579379 5 31 theme carbohydrate 708:719 arg1 sites 732:736 At least two carbohydrate attachment sites 695:736 At least two carbohydrate attachment sites 695:736 At least two carbohydrate attachment sites, one on each chain, have been identified. 2579379 7 32 theme complement 923:932 arg1 C4 944:945 complement component C4 923:945 complement component C4 923:945 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 1 33 theme complete 87:94 arg1 sequence 108:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 0 34 theme component 17:25 arg1 C3 27:28 Human complement component C3 0:28 Human complement component C3 0:28 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 6 35 theme nucleotide 825:834 arg1 level 836:840 the nucleotide level 821:840 the nucleotide level 821:840 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 4 36 theme mobilizing 674:683 arg1 activity 685:692 leukocyte mobilizing activity 664:692 leukocyte mobilizing activity 664:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 1 37 theme cDNA 96:99 arg1 sequence 108:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 2 38 theme alpha 320:324 arg1 chain 326:330 the alpha chain 316:330 the alpha chain (992 residues) 316:345 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 38 theme alpha 320:324 arg1 residues 337:344 992 residues 333:344 992 residues 333:344 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 38 theme alpha 320:324 arg1 residues 271:278 22 amino acid residues 257:278 22 amino acid residues 257:278 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 0 39 theme cDNA 31:34 arg1 sequence 43:50 cDNA coding sequence 31:50 cDNA coding sequence 31:50 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 4 40 with fragment 650:657 arg1 activity 685:692 leukocyte mobilizing activity 664:692 leukocyte mobilizing activity 664:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 0 41 dep sequence 43:50 arg1 C3 27:28 Human complement component C3 0:28 Human complement component C3 0:28 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 6 42 contain has 789:791 arg2 identity 797:804 79% identity 793:804 79% identity to mouse C3 at the nucleotide level 793:840 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 6 42 contain has 789:791 arg1 C3 786:787 Human C3 780:787 Human C3 780:787 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 6 42 contain has 789:791 arg2 identity 850:857 77% identity 846:857 77% identity at the amino acid level 846:881 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 6 43 from level 836:840 arg1 C3 815:816 mouse C3 809:816 mouse C3 at the nucleotide level 809:840 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 4 44 with sites 578:582 arg1 activity 685:692 leukocyte mobilizing activity 664:692 leukocyte mobilizing activity 664:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 45 with site 535:538 arg1 activity 685:692 leukocyte mobilizing activity 664:692 leukocyte mobilizing activity 664:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 3 46 theme arginine 382:389 arg1 residues 391:398 four arginine residues 377:398 four arginine residues not present in the mature protein 377:432 The two chains are joined by four arginine residues not present in the mature protein. 2579379 4 47 theme important 456:464 arg1 sites 466:470 Several functionally important sites 435:470 Several functionally important sites 435:470 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 7 48 theme protease 888:895 arg1 2-macroglobulin 903:917 protease alpha 2-macroglobulin 888:917 protease alpha 2-macroglobulin 888:917 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 2 49 theme precursor 209:217 arg1 molecule 219:226 The encoded precursor molecule 197:226 The encoded precursor molecule 197:226 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 1 50 theme coding 101:106 arg1 sequence 108:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence 83:115 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 0 51 theme coding 36:41 arg1 sequence 43:50 cDNA coding sequence 31:50 cDNA coding sequence 31:50 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 6 52 theme mouse 809:813 arg1 C3 815:816 mouse C3 809:816 mouse C3 at the nucleotide level 809:840 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 1 53 link derived 121:127 arg1 sequence 140:147 derived amino acid sequence 121:147 derived amino acid sequence of human complement component C3 121:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 2 54 theme encoded 201:207 arg1 molecule 219:226 The encoded precursor molecule 197:226 The encoded precursor molecule 197:226 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 0 55 link derived 56:62 arg1 structure 72:80 derived primary structure 56:80 derived primary structure 56:80 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 3 56 theme mature 419:424 arg1 protein 426:432 the mature protein 415:432 the mature protein 415:432 The two chains are joined by four arginine residues not present in the mature protein. 2579379 5 57 from one 739:741 arg1 chain 751:755 each chain 746:755 each chain 746:755 At least two carbohydrate attachment sites, one on each chain, have been identified. 2579379 3 58 theme present 404:410 arg1 residues 391:398 four arginine residues 377:398 four arginine residues not present in the mature protein 377:432 The two chains are joined by four arginine residues not present in the mature protein. 2579379 4 59 theme peptide 642:648 arg1 fragment 650:657 a peptide fragment 640:657 a peptide fragment with leukocyte mobilizing activity 640:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 60 theme cleavage 526:533 arg1 site 535:538 the cleavage site 522:538 the cleavage site liberating the anaphylatoxin 522:567 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 4 60 theme cleavage 526:533 arg1 cleavage 587:594 cleavage 587:594 cleavage by the serine protease factor I 587:626 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 7 61 theme component 934:942 arg1 C4 944:945 complement component C4 923:945 complement component C4 923:945 The protease alpha 2-macroglobulin and complement component C4 show considerable homology to C3, suggesting that the three proteins have evolved from a common ancestor. 2579379 4 62 with site 516:519 arg1 activity 685:692 leukocyte mobilizing activity 664:692 leukocyte mobilizing activity 664:692 Several functionally important sites have been localized, such as the thiolester site, the cleavage site liberating the anaphylatoxin, and two sites of cleavage by the serine protease factor I, as well as a peptide fragment with leukocyte mobilizing activity. 2579379 1 63 theme derived 121:127 arg1 sequence 140:147 derived amino acid sequence 121:147 derived amino acid sequence of human complement component C3 121:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 6 64 theme acid 872:875 arg1 level 877:881 the amino acid level 862:881 the amino acid level 862:881 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 0 65 theme derived 56:62 arg1 structure 72:80 derived primary structure 56:80 derived primary structure 56:80 Human complement component C3: cDNA coding sequence and derived primary structure. 2579379 2 66 theme signal 239:244 arg1 peptide 246:252 a signal peptide 237:252 a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues) 237:345 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 1 67 theme amino 129:133 arg1 sequence 140:147 derived amino acid sequence 121:147 derived amino acid sequence of human complement component C3 121:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 6 68 theme amino 866:870 arg1 level 877:881 the amino acid level 862:881 the amino acid level 862:881 Human C3 has 79% identity to mouse C3 at the nucleotide level and 77% identity at the amino acid level. 2579379 1 69 theme acid 135:138 arg1 sequence 140:147 derived amino acid sequence 121:147 derived amino acid sequence of human complement component C3 121:180 The complete cDNA coding sequence and derived amino acid sequence of human complement component C3 are presented. 2579379 2 70 contain contains 228:235 arg1 molecule 219:226 The encoded precursor molecule 197:226 The encoded precursor molecule 197:226 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 2579379 2 70 contain contains 228:235 arg2 peptide 246:252 a signal peptide 237:252 a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues) 237:345 The encoded precursor molecule contains a signal peptide of 22 amino acid residues, the beta chain (645 residues), and the alpha chain (992 residues). 17684013 0 0 theme new 75:77 arg1 mechanism 79:87 a new mechanism 73:87 a new mechanism of complement inhibition 73:112 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 6 1 theme stable 827:832 arg1 MG-ring 834:840 the structurally stable MG-ring 810:840 the structurally stable MG-ring formed by domains MG 1-6 810:865 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 0 2 with complex 27:33 arg1 C3c 61:63 complement component C3c 40:63 complement component C3c 40:63 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 1 3 theme major 152:156 arg1 activation 136:145 Undesired complement activation 115:145 Undesired complement activation 115:145 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 1 3 theme major 152:156 arg1 cause 158:162 a major cause 150:162 a major cause of tissue injury in various pathological conditions 150:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 1 4 theme complex 250:256 arg1 diseases 258:265 several immune complex diseases 235:265 several immune complex diseases 235:265 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 5 5 theme compstatin-binding 704:721 arg1 site 723:726 the compstatin-binding site 700:726 the compstatin-binding site 700:726 The structure reveals that the compstatin-binding site is formed by the macroglobulin (MG) domains 4 and 5. 17684013 6 6 theme binding 786:792 arg1 site 794:797 This binding site 781:797 This binding site 781:797 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 0 7 from Structure 0:8 arg1 complex 27:33 complex 27:33 complex with complement component C3c 27:63 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 1 8 from cause 158:162 arg1 conditions 205:214 various pathological conditions 184:214 various pathological conditions 184:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 2 9 theme 13-residue 282:291 arg1 peptide 293:299 a 13-residue peptide 280:299 a 13-residue peptide 280:299 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 2 9 theme 13-residue 282:291 arg1 Compstatin 268:277 Compstatin 268:277 Compstatin 268:277 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 2 10 theme complement 349:358 arg1 C3 370:371 complement component C3 349:371 complement component C3 349:371 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 6 11 theme known 898:902 arg1 site 912:915 any other known binding site 888:915 any other known binding site 888:915 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 0 12 theme inhibition 103:112 arg1 mechanism 79:87 a new mechanism 73:87 a new mechanism of complement inhibition 73:112 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 4 13 theme proteolytic 645:655 arg1 fragment 657:664 a major proteolytic fragment 637:664 a major proteolytic fragment of C3 637:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 4 13 theme proteolytic 645:655 arg1 C3c 632:634 C3c 632:634 C3c 632:634 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 2 14 theme crucial 403:409 arg1 step 411:414 a central and crucial step 389:414 a central and crucial step in the complement cascade 389:440 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 3 15 theme bound 496:500 arg1 form 502:505 the bound form 492:505 the bound form 492:505 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 8 16 theme substrate 1131:1139 arg1 C3 1141:1142 the substrate C3 1127:1142 the substrate C3 1127:1142 We propose a model in which compstatin sterically hinders the access of the substrate C3 to the convertase complexes, thus blocking complement activation and amplification. 17684013 4 17 theme compstatin 605:614 arg1 structure 592:600 the crystal structure 580:600 the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3 580:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 0 18 theme complement 92:101 arg1 inhibition 103:112 complement inhibition 92:112 complement inhibition 92:112 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 6 19 theme other 892:896 arg1 site 912:915 any other known binding site 888:915 any other known binding site 888:915 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 3 20 from site 463:466 arg1 C3 471:472 C3 471:472 C3 471:472 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 20 from site 463:466 arg1 form 502:505 the bound form 492:505 the bound form 492:505 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 5 21 theme macroglobulin 745:757 arg1 domains 764:770 the macroglobulin (MG) domains 4 and 5 741:778 the macroglobulin (MG) domains 4 and 5 741:778 The structure reveals that the compstatin-binding site is formed by the macroglobulin (MG) domains 4 and 5. 17684013 1 22 theme tissue 167:172 arg1 injury 174:179 tissue injury 167:179 tissue injury 167:179 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 2 23 theme effective 308:316 arg1 inhibitor 318:326 an effective inhibitor 305:326 an effective inhibitor of the activation of complement component C3 305:371 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 2 23 theme effective 308:316 arg1 Compstatin 268:277 Compstatin 268:277 Compstatin 268:277 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 1 24 theme injury 174:179 arg1 activation 136:145 Undesired complement activation 115:145 Undesired complement activation 115:145 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 1 24 theme injury 174:179 arg1 cause 158:162 a major cause 150:162 a major cause of tissue injury in various pathological conditions 150:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 2 25 theme complement 423:432 arg1 cascade 434:440 the complement cascade 419:440 the complement cascade 419:440 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 7 26 theme C3c 970:972 arg1 conformation 954:965 the conformation 950:965 the conformation of C3c 950:972 Compstatin does not alter the conformation of C3c, whereas compstatin itself undergoes a large conformational change upon binding. 17684013 4 27 theme C3 669:670 arg1 fragment 657:664 a major proteolytic fragment 637:664 a major proteolytic fragment of C3 637:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 4 27 theme C3 669:670 arg1 C3c 632:634 C3c 632:634 C3c 632:634 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 4 28 from structure 592:600 arg1 complex 619:625 complex 619:625 complex with C3c, a major proteolytic fragment of C3 619:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 3 29 theme binding 455:461 arg1 site 463:466 The precise binding site 443:466 The precise binding site on C3 443:472 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 29 theme binding 455:461 arg1 action 530:535 action 530:535 action of compstatin 530:549 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 29 theme binding 455:461 arg1 unknown 555:561 unknown 555:561 unknown 555:561 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 1 30 theme various 184:190 arg1 conditions 205:214 various pathological conditions 184:214 various pathological conditions 184:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 0 31 theme compstatin 13:22 arg1 Structure 0:8 Structure 0:8 Structure of compstatin in complex with complement component C3c 0:63 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 4 32 with complex 619:625 arg1 fragment 657:664 a major proteolytic fragment 637:664 a major proteolytic fragment of C3 637:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 4 32 with complex 619:625 arg1 C3c 632:634 C3c 632:634 C3c 632:634 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 3 33 theme exact 516:520 arg1 mode 522:525 the exact mode 512:525 the exact mode of action of compstatin 512:549 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 33 theme exact 516:520 arg1 unknown 555:561 unknown 555:561 unknown 555:561 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 1 34 theme pathological 192:203 arg1 conditions 205:214 various pathological conditions 184:214 various pathological conditions 184:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 4 35 theme major 639:643 arg1 fragment 657:664 a major proteolytic fragment 637:664 a major proteolytic fragment of C3 637:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 4 35 theme major 639:643 arg1 C3c 632:634 C3c 632:634 C3c 632:634 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 3 36 theme compstatin 540:549 arg1 action 530:535 action 530:535 action of compstatin 530:549 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 8 37 theme convertase 1151:1160 arg1 complexes 1162:1170 the convertase complexes 1147:1170 the convertase complexes 1147:1170 We propose a model in which compstatin sterically hinders the access of the substrate C3 to the convertase complexes, thus blocking complement activation and amplification. 17684013 6 38 theme binding 904:910 arg1 site 912:915 any other known binding site 888:915 any other known binding site 888:915 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 2 39 theme central 391:397 arg1 step 411:414 a central and crucial step 389:414 a central and crucial step in the complement cascade 389:440 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 0 40 theme component 51:59 arg1 C3c 61:63 complement component C3c 40:63 complement component C3c 40:63 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 8 41 theme C3 1141:1142 arg1 access 1117:1122 the access 1113:1122 the access of the substrate C3 to the convertase complexes 1113:1170 We propose a model in which compstatin sterically hinders the access of the substrate C3 to the convertase complexes, thus blocking complement activation and amplification. 17684013 6 42 dep domains 852:858 arg1 MG 860:861 MG 1-6 860:865 domains MG 1-6 852:865 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 3 43 theme action 530:535 arg1 structure 479:487 the structure 475:487 the structure in the bound form 475:505 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 43 theme action 530:535 arg1 site 463:466 The precise binding site 443:466 The precise binding site on C3 443:472 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 43 theme action 530:535 arg1 mode 522:525 the exact mode 512:525 the exact mode of action of compstatin 512:549 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 43 theme action 530:535 arg1 action 530:535 action 530:535 action of compstatin 530:549 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 43 theme action 530:535 arg1 unknown 555:561 unknown 555:561 unknown 555:561 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 2 44 theme activation 335:344 arg1 inhibitor 318:326 an effective inhibitor 305:326 an effective inhibitor of the activation of complement component C3 305:371 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 2 44 theme activation 335:344 arg1 Compstatin 268:277 Compstatin 268:277 Compstatin 268:277 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 0 45 theme complement 40:49 arg1 C3c 61:63 complement component C3c 40:63 complement component C3c 40:63 Structure of compstatin in complex with complement component C3c reveals a new mechanism of complement inhibition. 17684013 9 46 theme further 1264:1270 arg1 development 1272:1282 further development 1264:1282 further development of compstatin as a potential therapeutic 1264:1323 These insights are instrumental for further development of compstatin as a potential therapeutic. 17684013 1 47 theme Undesired 115:123 arg1 activation 136:145 Undesired complement activation 115:145 Undesired complement activation 115:145 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 1 47 theme Undesired 115:123 arg1 cause 158:162 a major cause 150:162 a major cause of tissue injury in various pathological conditions 150:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 7 48 theme conformational 1019:1032 arg1 change 1034:1039 a large conformational change 1011:1039 a large conformational change 1011:1039 Compstatin does not alter the conformation of C3c, whereas compstatin itself undergoes a large conformational change upon binding. 17684013 3 49 from structure 479:487 arg1 C3 471:472 C3 471:472 C3 471:472 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 49 from structure 479:487 arg1 form 502:505 the bound form 492:505 the bound form 492:505 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 1 50 theme complement 125:134 arg1 activation 136:145 Undesired complement activation 115:145 Undesired complement activation 115:145 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 1 50 theme complement 125:134 arg1 cause 158:162 a major cause 150:162 a major cause of tissue injury in various pathological conditions 150:214 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 8 51 theme complement 1187:1196 arg1 activation 1198:1207 complement activation 1187:1207 complement activation 1187:1207 We propose a model in which compstatin sterically hinders the access of the substrate C3 to the convertase complexes, thus blocking complement activation and amplification. 17684013 3 52 from mode 522:525 arg1 C3 471:472 C3 471:472 C3 471:472 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 52 from mode 522:525 arg1 form 502:505 the bound form 492:505 the bound form 492:505 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 9 53 theme compstatin 1287:1296 arg1 development 1272:1282 further development 1264:1282 further development of compstatin as a potential therapeutic 1264:1323 These insights are instrumental for further development of compstatin as a potential therapeutic. 17684013 4 54 theme crystal 584:590 arg1 structure 592:600 the crystal structure 580:600 the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3 580:670 Here we present the crystal structure of compstatin in complex with C3c, a major proteolytic fragment of C3. 17684013 3 55 theme precise 447:453 arg1 site 463:466 The precise binding site 443:466 The precise binding site on C3 443:472 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 55 theme precise 447:453 arg1 action 530:535 action 530:535 action of compstatin 530:549 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 3 55 theme precise 447:453 arg1 unknown 555:561 unknown 555:561 unknown 555:561 The precise binding site on C3, the structure in the bound form, and the exact mode of action of compstatin are unknown. 17684013 2 56 theme C3 370:371 arg1 activation 335:344 the activation 331:344 the activation of complement component C3 331:371 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 1 57 theme several 235:241 arg1 diseases 258:265 several immune complex diseases 235:265 several immune complex diseases 235:265 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 17684013 7 58 theme large 1013:1017 arg1 change 1034:1039 a large conformational change 1011:1039 a large conformational change 1011:1039 Compstatin does not alter the conformation of C3c, whereas compstatin itself undergoes a large conformational change upon binding. 17684013 6 59 theme MG-ring 834:840 arg1 part 802:805 part 802:805 part of the structurally stable MG-ring formed by domains MG 1-6 802:865 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 6 59 theme MG-ring 834:840 arg1 C3 920:921 C3 920:921 C3 920:921 This binding site is part of the structurally stable MG-ring formed by domains MG 1-6 and is far away from any other known binding site on C3. 17684013 5 60 theme MG 760:761 arg1 domains 764:770 the macroglobulin (MG) domains 4 and 5 741:778 the macroglobulin (MG) domains 4 and 5 741:778 The structure reveals that the compstatin-binding site is formed by the macroglobulin (MG) domains 4 and 5. 17684013 2 61 from step 411:414 arg1 cascade 434:440 the complement cascade 419:440 the complement cascade 419:440 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 2 62 theme component 360:368 arg1 C3 370:371 complement component C3 349:371 complement component C3 349:371 Compstatin, a 13-residue peptide, is an effective inhibitor of the activation of complement component C3 and thus blocks a central and crucial step in the complement cascade. 17684013 1 63 theme immune 243:248 arg1 diseases 258:265 several immune complex diseases 235:265 several immune complex diseases 235:265 Undesired complement activation is a major cause of tissue injury in various pathological conditions and contributes to several immune complex diseases. 19563754 1 0 theme receptor-mediated 149:165 arg1 endocytosis 167:177 receptor-mediated endocytosis 149:177 receptor-mediated endocytosis 149:177 LDL delivers cholesterol to lysosomes by receptor-mediated endocytosis. 19563754 3 1 theme isooctyl 326:333 arg1 chain 340:344 its isooctyl side chain 322:344 its isooctyl side chain buried 322:351 NPC2 binds cholesterol with its isooctyl side chain buried and its 3beta-hydroxyl exposed. 19563754 2 2 from lysosomes 205:213 arg1 Exit 180:183 Exit 180:183 Exit of cholesterol from lysosomes 180:213 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 7 3 theme cholesterol 932:942 arg1 exit 944:947 cholesterol exit 932:947 cholesterol exit from lysosomes 932:962 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 5 4 theme 3beta-hydroxyl 591:604 arg1 chain 631:635 3beta-hydroxyl buried and isooctyl side chain 591:635 3beta-hydroxyl buried and isooctyl side chain 591:635 NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3beta-hydroxyl buried and isooctyl side chain exposed. 19563754 7 5 theme distinct 840:847 arg1 subdomain 829:837 this subdomain 824:837 this subdomain (distinct from the binding subdomain) 824:875 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 8 6 theme working 1002:1008 arg1 model 1010:1014 a working model 1000:1014 a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes 1000:1244 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 5 7 theme buried 606:611 arg1 chain 631:635 3beta-hydroxyl buried and isooctyl side chain 591:635 3beta-hydroxyl buried and isooctyl side chain 591:635 NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3beta-hydroxyl buried and isooctyl side chain exposed. 19563754 4 8 theme NPC1 464:467 arg1 NTD 456:458 NTD 456:458 NTD 456:458 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 8 theme NPC1 464:467 arg1 NPC1 464:467 NPC1 464:467 NPC1 464:467 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 8 theme NPC1 464:467 arg1 domain 448:453 the N-terminal domain 433:453 the N-terminal domain (NTD) of NPC1 433:467 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 5 9 theme isooctyl 617:624 arg1 chain 631:635 3beta-hydroxyl buried and isooctyl side chain 591:635 3beta-hydroxyl buried and isooctyl side chain 591:635 NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3beta-hydroxyl buried and isooctyl side chain exposed. 19563754 4 10 theme high-resolution 403:417 arg1 structures 419:428 high-resolution structures 403:428 high-resolution structures of the N-terminal domain (NTD) of NPC1 403:467 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 2 11 theme soluble 280:286 arg1 NPC2 288:291 soluble NPC2 280:291 soluble NPC2 280:291 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 2 11 theme soluble 280:286 arg1 proteins 228:235 two proteins 224:235 two proteins 224:235 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 7 12 theme binding 858:864 arg1 subdomain 866:874 the binding subdomain 854:874 the binding subdomain 854:874 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 8 13 theme isooctyl 1191:1198 arg1 chain 1205:1209 its isooctyl side chain 1187:1209 its isooctyl side chain 1187:1209 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 0 14 theme cholesterol 95:105 arg1 binding 71:77 binding 71:77 binding 71:77 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 0 14 theme cholesterol 95:105 arg1 transfer 83:90 transfer 83:90 transfer 83:90 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 5 15 theme side 626:629 arg1 chain 631:635 3beta-hydroxyl buried and isooctyl side chain 591:635 3beta-hydroxyl buried and isooctyl side chain 591:635 NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3beta-hydroxyl buried and isooctyl side chain exposed. 19563754 4 16 with structures 419:428 arg1 25-hydroxycholesterol 504:524 25-hydroxycholesterol 504:524 25-hydroxycholesterol 504:524 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 16 with structures 419:428 arg1 cholesterol 488:498 cholesterol 488:498 cholesterol 488:498 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 8 17 theme chain 1205:1209 arg1 insertion 1174:1182 insertion 1174:1182 insertion of its isooctyl side chain into the outer lysosomal membranes 1174:1244 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 8 18 dep transfers 1108:1116 arg1 allowing 1165:1172 allowing 1165:1172 allowing insertion of its isooctyl side chain into the outer lysosomal membranes 1165:1244 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 8 18 dep transfers 1108:1116 arg1 reversing 1135:1143 reversing 1135:1143 reversing its orientation 1135:1159 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 5 19 theme opposite 573:580 arg1 orientation 561:571 an orientation 558:571 an orientation opposite to NPC2 558:588 NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3beta-hydroxyl buried and isooctyl side chain exposed. 19563754 7 20 from lysosomes 954:962 arg1 exit 944:947 cholesterol exit 932:947 cholesterol exit from lysosomes 932:962 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 8 21 theme lysosomal 1030:1038 arg1 hydrolysis 1040:1049 lysosomal hydrolysis 1030:1049 lysosomal hydrolysis of LDL-cholesteryl esters 1030:1075 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 6 22 from NPC2 672:675 arg1 transfer 658:665 Cholesterol transfer 646:665 Cholesterol transfer from NPC2 to NPC1(NTD) 646:688 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 8 23 theme side 1200:1203 arg1 chain 1205:1209 its isooctyl side chain 1187:1209 its isooctyl side chain 1187:1209 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 0 24 theme domain 24:29 arg1 Structure 0:8 Structure 0:8 Structure of N-terminal domain of NPC1 0:37 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 8 25 theme LDL-cholesteryl 1054:1068 arg1 esters 1070:1075 LDL-cholesteryl esters 1054:1075 LDL-cholesteryl esters 1054:1075 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 7 26 theme point 805:809 arg1 mutations 811:819 point mutations 805:819 point mutations in this subdomain (distinct from the binding subdomain) 805:875 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 0 27 theme N-terminal 13:22 arg1 NPC1 34:37 NPC1 34:37 NPC1 34:37 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 0 27 theme N-terminal 13:22 arg1 domain 24:29 N-terminal domain 13:29 N-terminal domain of NPC1 13:37 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 6 28 from reorientation 699:711 arg1 NPC1 739:742 NPC1 739:742 NPC1(NTD) 739:747 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 6 28 from reorientation 699:711 arg1 NTD 744:746 NTD 744:746 NTD 744:746 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 2 29 theme cholesterol 188:198 arg1 Exit 180:183 Exit 180:183 Exit of cholesterol from lysosomes 180:213 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 0 30 theme NPC1 34:37 arg1 NPC1 34:37 NPC1 34:37 NPC1 34:37 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 0 30 theme NPC1 34:37 arg1 domain 24:29 N-terminal domain 13:29 N-terminal domain of NPC1 13:37 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 7 31 with NPC1 795:798 arg1 mutations 811:819 point mutations 805:819 point mutations in this subdomain (distinct from the binding subdomain) 805:875 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 2 32 theme Niemann-Pick 253:264 arg1 proteins 228:235 two proteins 224:235 two proteins 224:235 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 2 32 theme Niemann-Pick 253:264 arg1 NPC1 270:273 NPC1 270:273 NPC1 270:273 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 2 32 theme Niemann-Pick 253:264 arg1 C1 266:267 membrane-bound Niemann-Pick C1 238:267 membrane-bound Niemann-Pick C1 (NPC1) 238:274 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 8 33 theme lysosomal 1226:1234 arg1 membranes 1236:1244 the outer lysosomal membranes 1216:1244 the outer lysosomal membranes 1216:1244 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 8 34 theme esters 1070:1075 arg1 hydrolysis 1040:1049 lysosomal hydrolysis 1030:1049 lysosomal hydrolysis of LDL-cholesteryl esters 1030:1075 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 2 35 theme membrane-bound 238:251 arg1 proteins 228:235 two proteins 224:235 two proteins 224:235 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 2 35 theme membrane-bound 238:251 arg1 NPC1 270:273 NPC1 270:273 NPC1 270:273 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 2 35 theme membrane-bound 238:251 arg1 C1 266:267 membrane-bound Niemann-Pick C1 238:267 membrane-bound Niemann-Pick C1 (NPC1) 238:274 Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. 19563754 8 36 theme outer 1220:1224 arg1 membranes 1236:1244 the outer lysosomal membranes 1216:1244 the outer lysosomal membranes 1216:1244 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 0 37 theme distinct 47:54 arg1 subdomains 56:65 distinct subdomains 47:65 distinct subdomains 47:65 Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. 19563754 8 38 dep model 1010:1014 arg1 wherein 1016:1022 wherein 1016:1022 wherein 1016:1022 We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes. 19563754 7 39 theme NPC1-deficient 967:980 arg1 cells 982:986 NPC1-deficient cells 967:986 NPC1-deficient cells 967:986 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 7 40 from mutations 811:819 arg1 subdomain 829:837 this subdomain 824:837 this subdomain (distinct from the binding subdomain) 824:875 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 6 41 theme cholesterol 776:786 arg1 entry 788:792 cholesterol entry 776:792 cholesterol entry 776:792 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 7 42 from subdomain 866:874 arg1 distinct 840:847 distinct 840:847 distinct 840:847 NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. 19563754 4 43 theme domain 448:453 arg1 complexes 473:481 complexes 473:481 complexes with cholesterol and 25-hydroxycholesterol 473:524 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 43 theme domain 448:453 arg1 structures 419:428 high-resolution structures 403:428 high-resolution structures of the N-terminal domain (NTD) of NPC1 403:467 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 6 44 theme subdomain 726:734 arg1 reorientation 699:711 reorientation 699:711 reorientation of a helical subdomain in NPC1(NTD) 699:747 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 6 45 theme Cholesterol 646:656 arg1 transfer 658:665 Cholesterol transfer 646:665 Cholesterol transfer from NPC2 to NPC1(NTD) 646:688 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 3 46 theme side 335:338 arg1 chain 340:344 its isooctyl side chain 322:344 its isooctyl side chain buried 322:351 NPC2 binds cholesterol with its isooctyl side chain buried and its 3beta-hydroxyl exposed. 19563754 4 47 theme N-terminal 437:446 arg1 NTD 456:458 NTD 456:458 NTD 456:458 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 47 theme N-terminal 437:446 arg1 NPC1 464:467 NPC1 464:467 NPC1 464:467 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 47 theme N-terminal 437:446 arg1 domain 448:453 the N-terminal domain 433:453 the N-terminal domain (NTD) of NPC1 433:467 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 6 48 theme helical 718:724 arg1 subdomain 726:734 a helical subdomain 716:734 a helical subdomain 716:734 Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. 19563754 4 49 with complexes 473:481 arg1 25-hydroxycholesterol 504:524 25-hydroxycholesterol 504:524 25-hydroxycholesterol 504:524 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 19563754 4 49 with complexes 473:481 arg1 cholesterol 488:498 cholesterol 488:498 cholesterol 488:498 Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. 3881423 9 0 theme purified 1817:1824 arg1 alpha-glucosidase 1840:1856 purified human neutral alpha-glucosidase A. 1817:1859 purified human neutral alpha-glucosidase A. 1817:1859 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 8 1 theme gel 1614:1616 arg1 electrophoresis 1618:1632 native gel electrophoresis 1607:1632 native gel electrophoresis 1607:1632 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 3 2 theme enzyme 532:537 arg1 glucosidase 539:549 the glycoprotein processing enzyme glucosidase II 504:552 the glycoprotein processing enzyme glucosidase II 504:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 9 3 theme neutral 1832:1838 arg1 alpha-glucosidase 1840:1856 purified human neutral alpha-glucosidase A. 1817:1859 purified human neutral alpha-glucosidase A. 1817:1859 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 4 4 theme alpha-glucosidase 669:685 arg1 AB 687:688 neutral alpha-glucosidase AB 661:688 neutral alpha-glucosidase AB 661:688 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 10 5 theme chromosome 2269:2278 arg1 arm 2256:2258 the long arm 2247:2258 the long arm of human chromosome II 2247:2281 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 10 5 theme chromosome 2269:2278 arg1 chromosome 2269:2278 human chromosome II 2263:2281 human chromosome II 2263:2281 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 5 6 theme cell 809:812 arg1 lines 814:818 both mutant and parental cell lines 784:818 both mutant and parental cell lines 784:818 In contrast, both mutant and parental cell lines exhibited several lysosomal hydrolases which are processed by glucosidase II. 3881423 10 7 theme long 2251:2254 arg1 arm 2256:2258 the long arm 2247:2258 the long arm of human chromosome II 2247:2281 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 10 7 theme long 2251:2254 arg1 chromosome 2269:2278 human chromosome II 2263:2281 human chromosome II 2263:2281 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 2 8 dep pH. 272:274 arg1 appears 289:295 appears 289:295 appears as a doublet of enzyme activity on native gel electrophoresis 289:357 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 2 8 dep pH. 272:274 arg1 termed 367:372 termed 367:372 was termed neutral alpha-glucosidase AB 363:401 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 6 9 theme neutral 979:985 arg1 AB 1005:1006 neutral alpha-glucosidase AB 979:1006 neutral alpha-glucosidase AB 979:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 9 10 theme neutral 2123:2129 arg1 AB 2149:2150 neutral alpha-glucosidase AB 2123:2150 neutral alpha-glucosidase AB 2123:2150 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 7 11 theme neutral 1168:1174 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 11 theme neutral 1168:1174 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 11 theme neutral 1168:1174 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 5 12 theme lysosomal 838:846 arg1 hydrolases 848:857 several lysosomal hydrolases 830:857 several lysosomal hydrolases which are processed by glucosidase II 830:895 In contrast, both mutant and parental cell lines exhibited several lysosomal hydrolases which are processed by glucosidase II. 3881423 9 13 theme subunit 1691:1697 arg1 size 1699:1702 a subunit size 1689:1702 a subunit size of 67 kDa 1689:1712 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 3 14 theme biochemical 430:440 arg1 evidence 442:449 genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II 418:552 genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II 418:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 8 15 theme native 1607:1612 arg1 electrophoresis 1618:1632 native gel electrophoresis 1607:1632 native gel electrophoresis 1607:1632 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 0 16 theme alpha-glucosidase 20:36 arg1 AB 38:39 neutral alpha-glucosidase AB 12:39 neutral alpha-glucosidase AB 12:39 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 6 17 theme alpha-glucosidase 946:962 arg1 component 966:974 the human neutral alpha-glucosidase A component 928:974 the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 928:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 9 18 theme alpha-glucosidase 1908:1924 arg1 AB 1926:1927 neutral alpha-glucosidase AB 1900:1927 neutral alpha-glucosidase AB 1900:1927 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 19 dep alpha-glucosidase 1840:1856 arg1 A. 1858:1859 A. 1858:1859 A. 1858:1859 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 6 20 theme human 932:936 arg1 component 966:974 the human neutral alpha-glucosidase A component 928:974 the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 928:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 9 21 theme kDa 1710:1712 arg1 size 1699:1702 a subunit size 1689:1702 a subunit size of 67 kDa 1689:1712 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 22 theme pig 1715:1717 arg1 glucosidase 1719:1729 pig glucosidase II 1715:1732 pig glucosidase II 1715:1732 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 7 23 theme anionic 1251:1257 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 23 theme anionic 1251:1257 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 23 theme anionic 1251:1257 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 4 24 from deficient 612:620 arg1 glucosidase 625:635 glucosidase II 625:638 glucosidase II 625:638 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 9 25 theme kDa 1779:1781 arg1 size 1767:1770 a subunit size 1757:1770 a subunit size of 100 kDa 1757:1781 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 6 26 theme AB 1005:1006 arg1 component 966:974 the human neutral alpha-glucosidase A component 928:974 the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 928:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 8 27 theme neutral 1505:1511 arg1 AB 1531:1532 human neutral alpha-glucosidase AB 1499:1532 human neutral alpha-glucosidase AB 1499:1532 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 7 28 contain have 1303:1306 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 28 contain have 1303:1306 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 28 contain have 1303:1306 arg2 optima 1319:1324 a broad pH optima 1308:1324 a broad pH optima (5.5-8.5) 1308:1334 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 28 contain have 1303:1306 arg2 5.5-8.5 1327:1333 5.5-8.5 1327:1333 5.5-8.5 1327:1333 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 28 contain have 1303:1306 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 9 29 theme glucosidase 1991:2001 arg1 substrate 1978:1986 the natural substrate 1966:1986 the natural substrate of glucosidase II 1966:2004 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 3 30 gly glycoprotein 508:519 arg1 glycoprotein 508:519 the glycoprotein processing enzyme glucosidase II 504:552 the glycoprotein processing enzyme glucosidase II 504:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 2 31 theme alpha-glucosidase 382:398 arg1 AB 400:401 neutral alpha-glucosidase AB 374:401 neutral alpha-glucosidase AB 374:401 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 7 32 theme high-molecular 1201:1214 arg1 mass 1216:1219 high-molecular mass 1201:1219 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 1 33 theme Biochemical 96:106 arg1 studies 120:126 Biochemical and genetic studies 96:126 Biochemical and genetic studies. 96:127 Biochemical and genetic studies. 3881423 8 34 theme purified 1535:1542 arg1 glucosidase 1548:1558 purified rat glucosidase II 1535:1561 purified rat glucosidase II 1535:1561 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 4 35 theme parental 760:767 arg1 parental 760:767 parental 760:767 parental 760:767 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 35 theme parental 760:767 arg1 % 755:755 less than 0.5% 742:755 less than 0.5% of parental 742:767 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 0 36 theme neutral 12:18 arg1 AB 38:39 neutral alpha-glucosidase AB 12:39 neutral alpha-glucosidase AB 12:39 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 1 37 theme genetic 112:118 arg1 studies 120:126 Biochemical and genetic studies 96:126 Biochemical and genetic studies. 96:127 Biochemical and genetic studies. 3881423 2 38 from pH. 272:274 arg1 active 254:259 active 254:259 active 254:259 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 2 38 from pH. 272:274 arg1 isozyme 216:222 a human isozyme 208:222 a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB 208:401 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 4 39 theme mutant 576:581 arg1 deficient 648:656 deficient 648:656 deficient 648:656 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 39 theme mutant 576:581 arg1 line 598:601 a mutant mouse lymphoma line 574:601 a mutant mouse lymphoma line which is deficient in glucosidase II 574:638 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 39 theme mutant 576:581 arg1 deficient 612:620 deficient 612:620 deficient 612:620 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 3 40 theme neutral 456:462 arg1 AB 482:483 neutral alpha-glucosidase AB 456:483 neutral alpha-glucosidase AB 456:483 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 0 41 theme AB 38:39 arg1 glucosidase 80:90 the glycoprotein processing enzyme glucosidase II 45:93 the glycoprotein processing enzyme glucosidase II 45:93 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 0 41 theme AB 38:39 arg1 Identity 0:7 Identity 0:7 Identity of neutral alpha-glucosidase AB 0:39 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 7 42 gly glycoproteins 1259:1271 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 42 gly glycoproteins 1259:1271 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 42 gly glycoproteins 1259:1271 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 43 theme broad 1310:1314 arg1 5.5-8.5 1327:1333 5.5-8.5 1327:1333 5.5-8.5 1327:1333 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 43 theme broad 1310:1314 arg1 optima 1319:1324 a broad pH optima 1308:1324 a broad pH optima (5.5-8.5) 1308:1334 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 44 dep mass 1216:1219 arg1 greater 1222:1228 greater 1222:1228 greater 1222:1228 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 4 45 from glucosidase 625:635 arg1 deficient 648:656 deficient 648:656 deficient 648:656 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 45 from glucosidase 625:635 arg1 line 598:601 a mutant mouse lymphoma line 574:601 a mutant mouse lymphoma line which is deficient in glucosidase II 574:638 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 45 from glucosidase 625:635 arg1 deficient 612:620 deficient 612:620 deficient 612:620 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 2 46 theme gel 339:341 arg1 electrophoresis 343:357 native gel electrophoresis 332:357 native gel electrophoresis 332:357 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 2 47 theme human 210:214 arg1 active 254:259 active 254:259 active 254:259 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 2 47 theme human 210:214 arg1 isozyme 216:222 a human isozyme 208:222 a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB 208:401 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 7 48 contain have 1341:1344 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 48 contain have 1341:1344 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 48 contain have 1341:1344 arg2 microM 1479:1484 35 versus 19 microM 1466:1484 35 versus 19 microM 1466:1484 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 48 contain have 1341:1344 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 48 contain have 1341:1344 arg2 Km 1356:1357 a similar Km 1346:1357 a similar Km for maltose (4.8 versus 2.1 mM) 1346:1389 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 48 contain have 1341:1344 arg2 4-methylumbelliferyl-alpha-D-glucopyranoside 1420:1463 the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside 1395:1463 the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1395:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 9 49 theme major 1800:1804 arg1 protein 1806:1812 the 98-kDa major protein 1789:1812 the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous 1789:2184 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 0 50 theme glycoprotein 49:60 arg1 glucosidase 80:90 the glycoprotein processing enzyme glucosidase II 45:93 the glycoprotein processing enzyme glucosidase II 45:93 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 0 51 gly glycoprotein 49:60 arg1 glycoprotein 49:60 the glycoprotein processing enzyme glucosidase II 45:93 the glycoprotein processing enzyme glucosidase II 45:93 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 6 52 theme pig 1117:1119 arg1 kidney 1121:1126 pig kidney 1117:1126 pig kidney 1117:1126 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 8 53 theme activity 1595:1602 arg1 doublet 1577:1583 a doublet 1575:1583 a doublet of enzyme activity 1575:1602 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 3 54 theme processing 521:530 arg1 glucosidase 539:549 the glycoprotein processing enzyme glucosidase II 504:552 the glycoprotein processing enzyme glucosidase II 504:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 9 55 theme rat 1644:1646 arg1 glucosidase 1648:1658 rat glucosidase II 1644:1661 rat glucosidase II 1644:1661 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 56 theme human 1826:1830 arg1 alpha-glucosidase 1840:1856 purified human neutral alpha-glucosidase A. 1817:1859 purified human neutral alpha-glucosidase A. 1817:1859 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 4 57 theme lymphoma 589:596 arg1 deficient 648:656 deficient 648:656 deficient 648:656 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 57 theme lymphoma 589:596 arg1 line 598:601 a mutant mouse lymphoma line 574:601 a mutant mouse lymphoma line which is deficient in glucosidase II 574:638 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 57 theme lymphoma 589:596 arg1 deficient 612:620 deficient 612:620 deficient 612:620 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 9 58 theme biochemical 2070:2080 arg1 synonymous 2175:2184 synonymous 2175:2184 synonymous 2175:2184 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 58 theme biochemical 2070:2080 arg1 data 2082:2085 the biochemical data 2066:2085 the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB 2066:2150 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 59 contain have 1752:1755 arg1 glucosidase 1719:1729 pig glucosidase II 1715:1732 pig glucosidase II 1715:1732 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 59 contain have 1752:1755 arg2 size 1767:1770 a subunit size 1757:1770 a subunit size of 100 kDa 1757:1781 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 7 60 theme artificial 1399:1408 arg1 microM 1479:1484 35 versus 19 microM 1466:1484 35 versus 19 microM 1466:1484 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 60 theme artificial 1399:1408 arg1 4-methylumbelliferyl-alpha-D-glucopyranoside 1420:1463 the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside 1395:1463 the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1395:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 9 61 theme consistent 2087:2096 arg1 synonymous 2175:2184 synonymous 2175:2184 synonymous 2175:2184 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 61 theme consistent 2087:2096 arg1 data 2082:2085 the biochemical data 2066:2085 the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB 2066:2150 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 3 62 with synonymous 488:497 arg1 glucosidase 539:549 the glycoprotein processing enzyme glucosidase II 504:552 the glycoprotein processing enzyme glucosidase II 504:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 4 63 from AB 687:688 arg1 deficient 648:656 deficient 648:656 deficient 648:656 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 63 from AB 687:688 arg1 line 598:601 a mutant mouse lymphoma line 574:601 a mutant mouse lymphoma line which is deficient in glucosidase II 574:638 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 63 from AB 687:688 arg1 deficient 612:620 deficient 612:620 deficient 612:620 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 7 64 theme substrate 1410:1418 arg1 microM 1479:1484 35 versus 19 microM 1466:1484 35 versus 19 microM 1466:1484 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 64 theme substrate 1410:1418 arg1 4-methylumbelliferyl-alpha-D-glucopyranoside 1420:1463 the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside 1395:1463 the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1395:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 4 65 theme neutral 661:667 arg1 AB 687:688 neutral alpha-glucosidase AB 661:688 neutral alpha-glucosidase AB 661:688 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 2 66 theme enzyme 313:318 arg1 activity 320:327 enzyme activity 313:327 enzyme activity 313:327 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 6 67 theme alpha-glucosidase 987:1003 arg1 AB 1005:1006 neutral alpha-glucosidase AB 979:1006 neutral alpha-glucosidase AB 979:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 9 68 from protein 1806:1812 arg1 alpha-glucosidase 1840:1856 purified human neutral alpha-glucosidase A. 1817:1859 purified human neutral alpha-glucosidase A. 1817:1859 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 4 69 from deficient 648:656 arg1 AB 687:688 neutral alpha-glucosidase AB 661:688 neutral alpha-glucosidase AB 661:688 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 5 70 theme several 830:836 arg1 hydrolases 848:857 several lysosomal hydrolases 830:857 several lysosomal hydrolases which are processed by glucosidase II 830:895 In contrast, both mutant and parental cell lines exhibited several lysosomal hydrolases which are processed by glucosidase II. 3881423 2 71 theme native 332:337 arg1 electrophoresis 343:357 native gel electrophoresis 332:357 native gel electrophoresis 332:357 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 6 72 theme A 964:964 arg1 component 966:974 the human neutral alpha-glucosidase A component 928:974 the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 928:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 10 73 theme human 2263:2267 arg1 chromosome 2269:2278 human chromosome II 2263:2281 human chromosome II 2263:2281 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 9 74 theme neutral 1900:1906 arg1 AB 1926:1927 neutral alpha-glucosidase AB 1900:1927 neutral alpha-glucosidase AB 1900:1927 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 2 75 theme alpha-glucosidase 227:243 arg1 active 254:259 active 254:259 active 254:259 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 2 75 theme alpha-glucosidase 227:243 arg1 isozyme 216:222 a human isozyme 208:222 a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB 208:401 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 6 76 theme neutral 938:944 arg1 component 966:974 the human neutral alpha-glucosidase A component 928:974 the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 928:1006 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 7 77 theme similar 1348:1354 arg1 Km 1356:1357 a similar Km 1346:1357 a similar Km for maltose (4.8 versus 2.1 mM) 1346:1389 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 0 78 theme enzyme 73:78 arg1 glucosidase 80:90 the glycoprotein processing enzyme glucosidase II 45:93 the glycoprotein processing enzyme glucosidase II 45:93 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 8 79 theme human 1499:1503 arg1 AB 1531:1532 human neutral alpha-glucosidase AB 1499:1532 human neutral alpha-glucosidase AB 1499:1532 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 10 80 theme These 2187:2191 arg1 results 2206:2212 These and previous results 2187:2212 results 2206:2212 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 9 81 dep protein 1806:1812 arg1 believe 2010:2016 believe 2010:2016 believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous 2010:2184 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 8 82 theme alpha-glucosidase 1513:1529 arg1 AB 1531:1532 human neutral alpha-glucosidase AB 1499:1532 human neutral alpha-glucosidase AB 1499:1532 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 9 83 theme natural 1970:1976 arg1 substrate 1978:1986 the natural substrate 1966:1986 the natural substrate of glucosidase II 1966:2004 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 2 84 theme activity 320:327 arg1 doublet 302:308 a doublet 300:308 a doublet of enzyme activity 300:327 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 7 85 theme mass 1216:1219 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 85 theme mass 1216:1219 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 85 theme mass 1216:1219 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 4 86 dep defined 694:700 arg1 parental 760:767 parental 760:767 parental 760:767 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 86 dep defined 694:700 arg1 % 755:755 less than 0.5% 742:755 less than 0.5% of parental 742:767 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 6 87 theme rat 1103:1105 arg1 liver 1107:1111 rat liver 1103:1111 rat liver 1103:1111 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 5 88 theme parental 800:807 arg1 lines 814:818 both mutant and parental cell lines 784:818 both mutant and parental cell lines 784:818 In contrast, both mutant and parental cell lines exhibited several lysosomal hydrolases which are processed by glucosidase II. 3881423 9 89 theme alpha-glucosidase 2131:2147 arg1 AB 2149:2150 neutral alpha-glucosidase AB 2123:2150 neutral alpha-glucosidase AB 2123:2150 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 90 theme subunit 1759:1765 arg1 size 1767:1770 a subunit size 1757:1770 a subunit size of 100 kDa 1757:1781 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 8 91 theme rat 1544:1546 arg1 glucosidase 1548:1558 purified rat glucosidase II 1535:1561 purified rat glucosidase II 1535:1561 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 10 92 theme previous 2197:2204 arg1 results 2206:2212 These and previous results 2187:2212 results 2206:2212 These and previous results would localize glucosidase II to the long arm of human chromosome II. 3881423 2 93 theme neutral 374:380 arg1 AB 400:401 neutral alpha-glucosidase AB 374:401 neutral alpha-glucosidase AB 374:401 We have previously partially purified, characterized, and chromosomally mapped a human isozyme of alpha-glucosidase which is active at neutral pH. This isozyme appears as a doublet of enzyme activity on native gel electrophoresis and was termed neutral alpha-glucosidase AB. 3881423 7 94 theme alpha-glucosidase 1176:1192 arg1 AB 1194:1195 neutral alpha-glucosidase AB 1168:1195 neutral alpha-glucosidase AB 1168:1195 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 94 theme alpha-glucosidase 1176:1192 arg1 glycoproteins 1259:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins 1201:1271 high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM) 1201:1485 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 94 theme alpha-glucosidase 1176:1192 arg1 glucosidase 1149:1159 glucosidase II 1149:1162 glucosidase II 1149:1162 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 95 theme pH 1316:1317 arg1 5.5-8.5 1327:1333 5.5-8.5 1327:1333 5.5-8.5 1327:1333 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 7 95 theme pH 1316:1317 arg1 optima 1319:1324 a broad pH optima 1308:1324 a broad pH optima (5.5-8.5) 1308:1334 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 4 96 theme mouse 583:587 arg1 deficient 648:656 deficient 648:656 deficient 648:656 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 96 theme mouse 583:587 arg1 line 598:601 a mutant mouse lymphoma line 574:601 a mutant mouse lymphoma line which is deficient in glucosidase II 574:638 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 4 96 theme mouse 583:587 arg1 deficient 612:620 deficient 612:620 deficient 612:620 We have found that a mutant mouse lymphoma line which is deficient in glucosidase II is also deficient in neutral alpha-glucosidase AB, as defined electrophoretically and quantitatively (less than 0.5% of parental). 3881423 6 97 theme biochemical 1055:1065 arg1 properties 1067:1076 its biochemical properties 1051:1076 its biochemical properties 1051:1076 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 3 98 theme genetic 418:424 arg1 evidence 442:449 genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II 418:552 genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II 418:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 6 99 dep liver 1107:1111 arg1 glucosidase 1128:1138 glucosidase II 1128:1141 glucosidase II 1128:1141 We have also further purified the human neutral alpha-glucosidase A component of neutral alpha-glucosidase AB 740-fold from placenta in order to compare its biochemical properties with those described for rat liver and pig kidney glucosidase II. 3881423 3 100 theme alpha-glucosidase 464:480 arg1 AB 482:483 neutral alpha-glucosidase AB 456:483 neutral alpha-glucosidase AB 456:483 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 5 101 theme mutant 789:794 arg1 lines 814:818 both mutant and parental cell lines 784:818 both mutant and parental cell lines 784:818 In contrast, both mutant and parental cell lines exhibited several lysosomal hydrolases which are processed by glucosidase II. 3881423 0 102 theme processing 62:71 arg1 glucosidase 80:90 the glycoprotein processing enzyme glucosidase II 45:93 the glycoprotein processing enzyme glucosidase II 45:93 Identity of neutral alpha-glucosidase AB and the glycoprotein processing enzyme glucosidase II. 3881423 9 103 theme genetic 2027:2033 arg1 evidence 2035:2042 the genetic evidence 2023:2042 the genetic evidence 2023:2042 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 103 theme genetic 2027:2033 arg1 compelling 2047:2056 compelling 2047:2056 compelling 2047:2056 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 3 104 theme glycoprotein 508:519 arg1 glucosidase 539:549 the glycoprotein processing enzyme glucosidase II 504:552 the glycoprotein processing enzyme glucosidase II 504:552 We now report genetic and biochemical evidence that neutral alpha-glucosidase AB is synonymous with the glycoprotein processing enzyme glucosidase II. 3881423 7 105 theme concanavalin 1287:1298 arg1 A 1300:1300 concanavalin A 1287:1300 concanavalin A 1287:1300 Both glucosidase II and neutral alpha-glucosidase AB are high-molecular mass (greater than 200,000 dalton) anionic glycoproteins which bind to concanavalin A, have a broad pH optima (5.5-8.5), and have a similar Km for maltose (4.8 versus 2.1 mM) and the artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (35 versus 19 microM). 3881423 9 106 theme 98-kDa 1793:1798 arg1 protein 1806:1812 the 98-kDa major protein 1789:1812 the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous 1789:2184 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 8 107 theme enzyme 1588:1593 arg1 activity 1595:1602 enzyme activity 1588:1602 enzyme activity 1588:1602 Similar to human neutral alpha-glucosidase AB, purified rat glucosidase II migrates as a doublet of enzyme activity on native gel electrophoresis. 3881423 9 108 contain have 1684:1687 arg1 glucosidase 1648:1658 rat glucosidase II 1644:1661 rat glucosidase II 1644:1661 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 108 contain have 1684:1687 arg2 size 1699:1702 a subunit size 1689:1702 a subunit size of 67 kDa 1689:1712 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 3881423 9 109 with consistent 2087:2096 arg1 hypothesis 2107:2116 the hypothesis 2103:2116 the hypothesis that neutral alpha-glucosidase AB 2103:2150 Although rat glucosidase II has been reported to have a subunit size of 67 kDa, pig glucosidase II has been found to have a subunit size of 100 kDa, like the 98-kDa major protein in purified human neutral alpha-glucosidase A. Although we have not demonstrated that neutral alpha-glucosidase AB is microsomal nor that it hydrolyzes the natural substrate of glucosidase II, we believe that the genetic evidence is compelling for and the biochemical data consistent with the hypothesis that neutral alpha-glucosidase AB and glucosidase II are synonymous. 23742080 8 0 theme Enzyme-linked 1114:1126 arg1 analysis 1140:1147 Enzyme-linked immunoassay analysis 1114:1147 Enzyme-linked immunoassay analysis of progranulin protein levels 1114:1177 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 0 1 theme protein 36:42 arg1 levels 44:49 TMEM106B protein levels 27:49 TMEM106B protein levels: implications for frontotemporal dementia 27:91 TMEM106B p.T185S regulates TMEM106B protein levels: implications for frontotemporal dementia. 23742080 2 2 theme intranuclear 281:292 arg1 inclusions 294:303 neuronal cytoplasmic or intranuclear inclusions 257:303 neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 257:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 8 3 theme progranulin 1152:1162 arg1 levels 1172:1177 progranulin protein levels 1152:1177 progranulin protein levels 1152:1177 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 4 4 theme progranulin 655:665 arg1 mutations 673:681 progranulin (GRN) mutations 655:681 progranulin (GRN) mutations 655:681 Intriguingly, the most significant association was in FTLD-TDP patients carrying progranulin (GRN) mutations. 23742080 7 5 theme TMEM106B-specific 1052:1068 arg1 antibody 1070:1077 a TMEM106B-specific antibody 1050:1077 a TMEM106B-specific antibody for investigation of this protein 1050:1111 We next generated and characterized a TMEM106B-specific antibody for investigation of this protein. 23742080 2 6 theme cytoplasmic 266:276 arg1 inclusions 294:303 neuronal cytoplasmic or intranuclear inclusions 257:303 neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 257:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 8 7 theme levels 1172:1177 arg1 analysis 1140:1147 Enzyme-linked immunoassay analysis 1114:1147 Enzyme-linked immunoassay analysis of progranulin protein levels 1114:1177 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 4 8 theme GRN 668:670 arg1 mutations 673:681 progranulin (GRN) mutations 655:681 progranulin (GRN) mutations 655:681 Intriguingly, the most significant association was in FTLD-TDP patients carrying progranulin (GRN) mutations. 23742080 13 9 theme cell 1793:1796 arg1 studies 1806:1812 Our cell culture studies 1789:1812 Our cell culture studies 1789:1812 Our cell culture studies provide evidence that the protective S185 isoform is degraded more rapidly than T185 TMEM106B, potentially due to differences in glycosylation. 23742080 2 10 theme TAR 322:324 arg1 protein 338:344 ubiquitinated TAR DNA binding protein 43 308:347 ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 308:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 2 10 theme TAR 322:324 arg1 FTLD-TDP 350:357 FTLD-TDP 350:357 FTLD-TDP 350:357 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 8 11 theme similar 1186:1192 arg1 effects 1194:1200 similar effects 1186:1200 similar effects 1186:1200 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 5 12 with disequilibrium 772:785 arg1 rs1990622 792:800 rs1990622 792:800 rs1990622 792:800 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 5 13 theme progranulin 811:821 arg1 biology 838:844 progranulin protein (PGRN) biology 811:844 progranulin protein (PGRN) biology 811:844 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 10 14 from differences 1452:1462 arg1 residue 1486:1492 residue N183 1486:1497 residue N183 1486:1497 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 10 14 from differences 1452:1462 arg1 N-glycosylation 1467:1481 N-glycosylation 1467:1481 N-glycosylation at residue N183 1467:1497 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 8 15 link Enzyme-linked 1114:1126 arg1 analysis 1140:1147 Enzyme-linked immunoassay analysis 1114:1147 Enzyme-linked immunoassay analysis of progranulin protein levels 1114:1177 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 5 16 from rs3173615 741:749 arg1 disequilibrium 772:785 linkage disequilibrium 764:785 linkage disequilibrium with rs1990622 764:800 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 2 17 theme many 207:210 arg1 patients 212:219 many patients 207:219 many patients 207:219 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 10 18 from residue 1486:1492 arg1 differences 1452:1462 differences 1452:1462 differences in N-glycosylation at residue N183 1452:1497 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 10 18 from residue 1486:1492 arg1 N-glycosylation 1467:1481 N-glycosylation 1467:1481 N-glycosylation at residue N183 1467:1497 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 11 19 from differences 1587:1597 arg1 risk 1611:1614 FTLD-TDP risk 1602:1614 FTLD-TDP risk 1602:1614 Together, our results provide a potential mechanism by which TMEM106B variants lead to differences in FTLD-TDP risk. 23742080 6 20 theme new 977:979 arg1 cohort 981:986 a new cohort 975:986 a new cohort of GRN mutation carriers 975:1011 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 9 21 theme T185 1275:1278 arg1 over-expression 1256:1270 over-expression 1256:1270 over-expression of T185 1256:1278 However, over-expression of T185 consistently led to higher TMEM106B protein levels than S185. 23742080 2 22 theme predominant 226:236 arg1 pathology 238:246 the predominant pathology 222:246 the predominant pathology 222:246 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 5 23 theme protein 864:870 arg1 TMEM106B 879:886 TMEM106B 879:886 TMEM106B 879:886 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 5 23 theme protein 864:870 arg1 B 876:876 transmembrane protein 106 B 850:876 transmembrane protein 106 B (TMEM106B) regulation 850:898 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 12 24 theme TAR 1719:1721 arg1 protein 1735:1741 TAR DNA binding protein 43 1719:1744 TAR DNA binding protein 43 pathology caused by progranulin mutations 1719:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 11 25 theme potential 1532:1540 arg1 mechanism 1542:1550 a potential mechanism 1530:1550 a potential mechanism by which TMEM106B variants lead to differences in FTLD-TDP risk 1530:1614 Together, our results provide a potential mechanism by which TMEM106B variants lead to differences in FTLD-TDP risk. 23742080 1 26 from cause 157:161 arg1 individuals 178:188 individuals 178:188 individuals under age 65 178:201 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 5 27 theme B 876:876 arg1 regulation 889:898 transmembrane protein 106 B (TMEM106B) regulation 850:898 transmembrane protein 106 B (TMEM106B) regulation 850:898 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 12 28 theme binding 1727:1733 arg1 protein 1735:1741 TAR DNA binding protein 43 1719:1744 TAR DNA binding protein 43 pathology caused by progranulin mutations 1719:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 9 29 theme higher 1300:1305 arg1 levels 1324:1329 higher TMEM106B protein levels 1300:1329 higher TMEM106B protein levels than S185 1300:1339 However, over-expression of T185 consistently led to higher TMEM106B protein levels than S185. 23742080 6 30 theme TMEM106B 940:947 arg1 variants 949:956 TMEM106B variants 940:956 TMEM106B variants 940:956 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 3 31 theme risk 441:444 arg1 factor 446:451 the first FTLD-TDP genetic risk factor 414:451 the first FTLD-TDP genetic risk factor 414:451 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 5 32 theme coding 725:730 arg1 rs3173615 741:749 rs3173615 741:749 rs3173615 (p.T185S) in linkage disequilibrium with rs1990622 741:800 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 5 32 theme coding 725:730 arg1 variant 732:738 the coding variant 721:738 the coding variant 721:738 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 9 33 theme protein 1316:1322 arg1 levels 1324:1329 higher TMEM106B protein levels 1300:1329 higher TMEM106B protein levels than S185 1300:1339 However, over-expression of T185 consistently led to higher TMEM106B protein levels than S185. 23742080 3 34 theme genome-wide 373:383 arg1 study 397:401 a genome-wide association study 371:401 a genome-wide association study 371:401 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 3 35 theme FTLD-TDP 424:431 arg1 factor 446:451 the first FTLD-TDP genetic risk factor 414:451 the first FTLD-TDP genetic risk factor 414:451 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 0 36 theme TMEM106B 0:7 arg1 p.T185S 9:15 TMEM106B p.T185S 0:15 TMEM106B p.T185S 0:15 TMEM106B p.T185S regulates TMEM106B protein levels: implications for frontotemporal dementia. 23742080 1 37 theme lobar 109:113 arg1 cause 157:161 the second leading cause 138:161 the second leading cause of dementia in individuals under age 65 138:201 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 37 theme lobar 109:113 arg1 FTLD 129:132 FTLD 129:132 FTLD 129:132 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 37 theme lobar 109:113 arg1 degeneration 115:126 Frontotemporal lobar degeneration 94:126 Frontotemporal lobar degeneration (FTLD) 94:133 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 12 38 theme genetic 1649:1655 arg1 variant 1657:1663 the p.T185S TMEM106B genetic variant 1628:1663 the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations 1628:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 6 39 theme mutation 995:1002 arg1 carriers 1004:1011 GRN mutation carriers 991:1011 GRN mutation carriers 991:1011 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 1 40 theme dementia 166:173 arg1 degeneration 115:126 Frontotemporal lobar degeneration 94:126 Frontotemporal lobar degeneration (FTLD) 94:133 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 40 theme dementia 166:173 arg1 cause 157:161 the second leading cause 138:161 the second leading cause of dementia in individuals under age 65 138:201 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 10 41 dep due 1445:1447 arg1 faster 1406:1411 faster 1406:1411 faster 1406:1411 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 5 42 theme linkage 764:770 arg1 disequilibrium 772:785 linkage disequilibrium 764:785 linkage disequilibrium with rs1990622 764:800 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 11 43 theme FTLD-TDP 1602:1609 arg1 risk 1611:1614 FTLD-TDP risk 1602:1614 FTLD-TDP risk 1602:1614 Together, our results provide a potential mechanism by which TMEM106B variants lead to differences in FTLD-TDP risk. 23742080 2 44 theme protein 338:344 arg1 inclusions 294:303 neuronal cytoplasmic or intranuclear inclusions 257:303 neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 257:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 14 45 theme low 1986:1988 arg1 levels 1999:2004 low TMEM106B levels 1986:2004 low TMEM106B levels 1986:2004 These findings suggest that low TMEM106B levels might protect against FTLD-TDP in these patients. 23742080 1 46 theme leading 149:155 arg1 degeneration 115:126 Frontotemporal lobar degeneration 94:126 Frontotemporal lobar degeneration (FTLD) 94:133 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 46 theme leading 149:155 arg1 cause 157:161 the second leading cause 138:161 the second leading cause of dementia in individuals under age 65 138:201 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 10 47 gly N-glycosylation 1467:1481 arg2 residue 1486:1492 residue N183 1486:1497 residue N183 1486:1497 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 10 47 gly N-glycosylation 1467:1481 arg1 residue 1486:1492 residue N183 1486:1497 residue N183 1486:1497 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 13 48 theme protective 1840:1849 arg1 isoform 1856:1862 the protective S185 isoform 1836:1862 the protective S185 isoform 1836:1862 Our cell culture studies provide evidence that the protective S185 isoform is degraded more rapidly than T185 TMEM106B, potentially due to differences in glycosylation. 23742080 4 49 theme significant 597:607 arg1 association 609:619 the most significant association 588:619 the most significant association 588:619 Intriguingly, the most significant association was in FTLD-TDP patients carrying progranulin (GRN) mutations. 23742080 10 50 theme treatment 1356:1364 arg1 experiments 1366:1376 Cycloheximide treatment experiments 1342:1376 Cycloheximide treatment experiments 1342:1376 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 3 51 theme TMEM106B 490:497 arg1 gene 499:502 the TMEM106B gene 486:502 the TMEM106B gene (top SNP rs1990622) 486:522 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 3 51 theme TMEM106B 490:497 arg1 SNP 509:511 top SNP rs1990622 505:521 top SNP rs1990622 505:521 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 11 52 theme TMEM106B 1561:1568 arg1 variants 1570:1577 TMEM106B variants 1561:1577 TMEM106B variants 1561:1577 Together, our results provide a potential mechanism by which TMEM106B variants lead to differences in FTLD-TDP risk. 23742080 8 53 theme immunoassay 1128:1138 arg1 analysis 1140:1147 Enzyme-linked immunoassay analysis 1114:1147 Enzyme-linked immunoassay analysis of progranulin protein levels 1114:1177 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 5 54 theme PGRN 832:835 arg1 biology 838:844 progranulin protein (PGRN) biology 811:844 progranulin protein (PGRN) biology 811:844 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 7 55 theme protein 1105:1111 arg1 investigation 1083:1095 investigation 1083:1095 investigation of this protein 1083:1111 We next generated and characterized a TMEM106B-specific antibody for investigation of this protein. 23742080 12 56 theme progranulin 1766:1776 arg1 mutations 1778:1786 progranulin mutations 1766:1786 progranulin mutations 1766:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 6 57 with association 925:935 arg1 FTLD-TDP 963:970 FTLD-TDP 963:970 FTLD-TDP 963:970 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 2 58 theme DNA 326:328 arg1 protein 338:344 ubiquitinated TAR DNA binding protein 43 308:347 ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 308:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 2 58 theme DNA 326:328 arg1 FTLD-TDP 350:357 FTLD-TDP 350:357 FTLD-TDP 350:357 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 8 59 theme protein 1164:1170 arg1 levels 1172:1177 progranulin protein levels 1152:1177 progranulin protein levels 1152:1177 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 13 60 theme culture 1798:1804 arg1 studies 1806:1812 Our cell culture studies 1789:1812 Our cell culture studies 1789:1812 Our cell culture studies provide evidence that the protective S185 isoform is degraded more rapidly than T185 TMEM106B, potentially due to differences in glycosylation. 23742080 2 61 theme ubiquitinated 308:320 arg1 protein 338:344 ubiquitinated TAR DNA binding protein 43 308:347 ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 308:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 2 61 theme ubiquitinated 308:320 arg1 FTLD-TDP 350:357 FTLD-TDP 350:357 FTLD-TDP 350:357 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 3 62 theme FTLD-TDP 559:566 arg1 risk 568:571 FTLD-TDP risk 559:571 FTLD-TDP risk 559:571 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 12 63 theme p.T185S 1632:1638 arg1 variant 1657:1663 the p.T185S TMEM106B genetic variant 1628:1663 the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations 1628:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 1 64 theme second 142:147 arg1 degeneration 115:126 Frontotemporal lobar degeneration 94:126 Frontotemporal lobar degeneration (FTLD) 94:133 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 64 theme second 142:147 arg1 cause 157:161 the second leading cause 138:161 the second leading cause of dementia in individuals under age 65 138:201 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 5 65 theme protein 823:829 arg1 biology 838:844 progranulin protein (PGRN) biology 811:844 progranulin protein (PGRN) biology 811:844 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 4 66 theme FTLD-TDP 628:635 arg1 patients 637:644 FTLD-TDP patients 628:644 FTLD-TDP patients carrying progranulin (GRN) mutations 628:681 Intriguingly, the most significant association was in FTLD-TDP patients carrying progranulin (GRN) mutations. 23742080 2 67 theme neuronal 257:264 arg1 inclusions 294:303 neuronal cytoplasmic or intranuclear inclusions 257:303 neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 257:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 12 68 theme protein 1735:1741 arg1 pathology 1746:1754 TAR DNA binding protein 43 pathology 1719:1754 TAR DNA binding protein 43 pathology caused by progranulin mutations 1719:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 5 69 theme transmembrane 850:862 arg1 TMEM106B 879:886 TMEM106B 879:886 TMEM106B 879:886 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 5 69 theme transmembrane 850:862 arg1 B 876:876 transmembrane protein 106 B 850:876 transmembrane protein 106 B (TMEM106B) regulation 850:898 Here, we investigated to what extent the coding variant, rs3173615 (p.T185S) in linkage disequilibrium with rs1990622, affects progranulin protein (PGRN) biology and transmembrane protein 106 B (TMEM106B) regulation. 23742080 13 70 from differences 1928:1938 arg1 glycosylation 1943:1955 glycosylation 1943:1955 glycosylation 1943:1955 Our cell culture studies provide evidence that the protective S185 isoform is degraded more rapidly than T185 TMEM106B, potentially due to differences in glycosylation. 23742080 12 71 theme DNA 1723:1725 arg1 protein 1735:1741 TAR DNA binding protein 43 1719:1744 TAR DNA binding protein 43 pathology caused by progranulin mutations 1719:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 6 72 theme variants 949:956 arg1 association 925:935 the association 921:935 the association of TMEM106B variants with FTLD-TDP 921:970 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 9 73 theme TMEM106B 1307:1314 arg1 levels 1324:1329 higher TMEM106B protein levels 1300:1329 higher TMEM106B protein levels than S185 1300:1339 However, over-expression of T185 consistently led to higher TMEM106B protein levels than S185. 23742080 1 74 theme Frontotemporal 94:107 arg1 cause 157:161 the second leading cause 138:161 the second leading cause of dementia in individuals under age 65 138:201 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 74 theme Frontotemporal 94:107 arg1 FTLD 129:132 FTLD 129:132 FTLD 129:132 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 1 74 theme Frontotemporal 94:107 arg1 degeneration 115:126 Frontotemporal lobar degeneration 94:126 Frontotemporal lobar degeneration (FTLD) 94:133 Frontotemporal lobar degeneration (FTLD) is the second leading cause of dementia in individuals under age 65. 23742080 3 75 theme association 385:395 arg1 study 397:401 a genome-wide association study 371:401 a genome-wide association study 371:401 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 3 76 theme genetic 433:439 arg1 factor 446:451 the first FTLD-TDP genetic risk factor 414:451 the first FTLD-TDP genetic risk factor 414:451 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 0 77 theme TMEM106B 27:34 arg1 levels 44:49 TMEM106B protein levels 27:49 TMEM106B protein levels: implications for frontotemporal dementia 27:91 TMEM106B p.T185S regulates TMEM106B protein levels: implications for frontotemporal dementia. 23742080 12 78 theme TMEM106B 1640:1647 arg1 variant 1657:1663 the p.T185S TMEM106B genetic variant 1628:1663 the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations 1628:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 6 79 theme carriers 1004:1011 arg1 cohort 981:986 a new cohort 975:986 a new cohort of GRN mutation carriers 975:1011 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 8 80 dep T185 1207:1210 arg1 over-expression 1230:1244 over-expression 1230:1244 over-expression 1230:1244 Enzyme-linked immunoassay analysis of progranulin protein levels showed similar effects upon T185 and S185 TMEM106B over-expression. 23742080 12 81 with dementia 1705:1712 arg1 pathology 1746:1754 TAR DNA binding protein 43 pathology 1719:1754 TAR DNA binding protein 43 pathology caused by progranulin mutations 1719:1786 We studied the p.T185S TMEM106B genetic variant previously implicated in frontotemporal dementia with TAR DNA binding protein 43 pathology caused by progranulin mutations. 23742080 6 82 theme GRN 991:993 arg1 carriers 1004:1011 GRN mutation carriers 991:1011 GRN mutation carriers 991:1011 First, we confirmed the association of TMEM106B variants with FTLD-TDP in a new cohort of GRN mutation carriers. 23742080 3 83 theme first 418:422 arg1 factor 446:451 the first FTLD-TDP genetic risk factor 414:451 the first FTLD-TDP genetic risk factor 414:451 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 10 84 theme Cycloheximide 1342:1354 arg1 experiments 1366:1376 Cycloheximide treatment experiments 1342:1376 Cycloheximide treatment experiments 1342:1376 Cycloheximide treatment experiments revealed that S185 degrades faster than T185 TMEM106B, potentially due to differences in N-glycosylation at residue N183. 23742080 0 85 dep levels 44:49 arg1 implications 52:63 implications 52:63 TMEM106B protein levels: implications for frontotemporal dementia 27:91 TMEM106B p.T185S regulates TMEM106B protein levels: implications for frontotemporal dementia. 23742080 14 86 theme TMEM106B 1990:1997 arg1 levels 1999:2004 low TMEM106B levels 1986:2004 low TMEM106B levels 1986:2004 These findings suggest that low TMEM106B levels might protect against FTLD-TDP in these patients. 23742080 4 87 contain carrying 646:653 arg1 patients 637:644 FTLD-TDP patients 628:644 FTLD-TDP patients carrying progranulin (GRN) mutations 628:681 Intriguingly, the most significant association was in FTLD-TDP patients carrying progranulin (GRN) mutations. 23742080 4 87 contain carrying 646:653 arg2 mutations 673:681 progranulin (GRN) mutations 655:681 progranulin (GRN) mutations 655:681 Intriguingly, the most significant association was in FTLD-TDP patients carrying progranulin (GRN) mutations. 23742080 2 88 theme binding 330:336 arg1 protein 338:344 ubiquitinated TAR DNA binding protein 43 308:347 ubiquitinated TAR DNA binding protein 43 (FTLD-TDP) 308:358 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 2 88 theme binding 330:336 arg1 FTLD-TDP 350:357 FTLD-TDP 350:357 FTLD-TDP 350:357 In many patients, the predominant pathology includes neuronal cytoplasmic or intranuclear inclusions of ubiquitinated TAR DNA binding protein 43 (FTLD-TDP). 23742080 13 89 theme S185 1851:1854 arg1 isoform 1856:1862 the protective S185 isoform 1836:1862 the protective S185 isoform 1836:1862 Our cell culture studies provide evidence that the protective S185 isoform is degraded more rapidly than T185 TMEM106B, potentially due to differences in glycosylation. 23742080 3 90 theme top 505:507 arg1 gene 499:502 the TMEM106B gene 486:502 the TMEM106B gene (top SNP rs1990622) 486:522 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 23742080 3 90 theme top 505:507 arg1 SNP 509:511 top SNP rs1990622 505:521 top SNP rs1990622 505:521 Recently, a genome-wide association study identified the first FTLD-TDP genetic risk factor, in which variants in and around the TMEM106B gene (top SNP rs1990622) were significantly associated with FTLD-TDP risk. 26416729 5 0 theme antagonists 796:806 arg1 recognition 781:791 the molecular recognition 767:791 the molecular recognition of antagonists 767:806 Our structure allows us to explore in detail the molecular recognition of antagonists. 26416729 2 1 theme agonists 402:409 arg1 binding 391:397 the binding 387:397 the binding of agonists 387:409 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 6 2 theme ion 1041:1043 arg1 pore 1045:1048 the ion pore 1037:1048 the ion pore 1037:1048 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 6 3 theme site 974:977 arg1 expansion 937:945 an expansion 934:945 an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 934:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 1 4 theme fast 174:177 arg1 neurotransmission 179:195 fast neurotransmission 174:195 fast neurotransmission 174:195 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 6 5 theme pore 1045:1048 arg1 closure 1026:1032 closure 1026:1032 closure of the ion pore in the transmembrane domain 1026:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 6 6 theme binding 966:972 arg1 site 974:977 the orthosteric binding site 950:977 the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 950:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 2 7 theme eukaryotic 324:333 arg1 receptors 344:352 prokaryotic and eukaryotic Cys-loop receptors 308:352 prokaryotic and eukaryotic Cys-loop receptors 308:352 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 4 8 theme high-specificity 681:696 arg1 affinity 671:678 a high affinity 664:678 a high affinity 664:678 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 8 theme high-specificity 681:696 arg1 antagonist 698:707 high-specificity antagonist 681:707 high-specificity antagonist 681:707 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 9 theme glycine 615:621 arg1 homopentamer 635:646 the human glycine receptor-α3 homopentamer 605:646 the human glycine receptor-α3 homopentamer 605:646 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 2 10 theme tremendous 362:371 arg1 insights 373:380 tremendous insights 362:380 tremendous insights into the binding of agonists 362:409 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 6 11 theme orthosteric 954:964 arg1 site 974:977 the orthosteric binding site 950:977 the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 950:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 4 12 theme human 609:613 arg1 homopentamer 635:646 the human glycine receptor-α3 homopentamer 605:646 the human glycine receptor-α3 homopentamer 605:646 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 3 13 theme inactivation 517:528 arg1 mechanism 504:512 the mechanism 500:512 the mechanism of inactivation by antagonists 500:543 Yet the mechanism of inactivation by antagonists remains unknown. 26416729 2 14 theme prokaryotic 308:318 arg1 receptors 344:352 prokaryotic and eukaryotic Cys-loop receptors 308:352 prokaryotic and eukaryotic Cys-loop receptors 308:352 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 1 15 theme neurotransmission 179:195 arg1 mediators 161:169 essential mediators 151:169 essential mediators of fast neurotransmission 151:195 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 1 15 theme neurotransmission 179:195 arg1 channels 106:113 Neurotransmitter-gated ion channels 79:113 Neurotransmitter-gated ion channels of the Cys-loop receptor family 79:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 5 16 theme molecular 771:779 arg1 recognition 781:791 the molecular recognition 767:791 the molecular recognition of antagonists 767:806 Our structure allows us to explore in detail the molecular recognition of antagonists. 26416729 1 17 theme Neurotransmitter-gated 79:100 arg1 channels 106:113 Neurotransmitter-gated ion channels 79:113 Neurotransmitter-gated ion channels of the Cys-loop receptor family 79:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 1 17 theme Neurotransmitter-gated 79:100 arg1 mediators 161:169 essential mediators 151:169 essential mediators of fast neurotransmission 151:195 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 2 18 theme channel 476:482 arg1 activation 484:493 channel activation 476:493 channel activation 476:493 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 6 19 theme previous 826:833 arg1 structures 835:844 previous structures 826:844 previous structures 826:844 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 1 20 theme ion 102:104 arg1 channels 106:113 Neurotransmitter-gated ion channels 79:113 Neurotransmitter-gated ion channels of the Cys-loop receptor family 79:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 1 20 theme ion 102:104 arg1 mediators 161:169 essential mediators 151:169 essential mediators of fast neurotransmission 151:195 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 0 21 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human glycine receptor-α3 0:45 Crystal structure of human glycine receptor-α3 bound to antagonist strychnine. 26416729 4 22 theme Å 584:584 arg1 structure 592:600 a 3.0 Å X-ray structure 578:600 a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine 578:719 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 6 23 theme antagonist-induced 869:886 arg1 inactivation 888:899 antagonist-induced inactivation 869:899 antagonist-induced inactivation of Cys-loop receptors 869:921 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 1 24 theme nervous 212:218 arg1 system 220:225 the nervous system 208:225 the nervous system 208:225 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 0 25 theme human 21:25 arg1 receptor-α3 35:45 human glycine receptor-α3 21:45 human glycine receptor-α3 21:45 Crystal structure of human glycine receptor-α3 bound to antagonist strychnine. 26416729 2 26 theme receptors 344:352 arg1 structures 294:303 Available X-ray structures 278:303 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors 278:352 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 6 27 with Comparisons 809:819 arg1 structures 835:844 previous structures 826:844 previous structures 826:844 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 2 28 theme Cys-loop 335:342 arg1 receptors 344:352 prokaryotic and eukaryotic Cys-loop receptors 308:352 prokaryotic and eukaryotic Cys-loop receptors 308:352 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 0 29 theme receptor-α3 35:45 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of human glycine receptor-α3 0:45 Crystal structure of human glycine receptor-α3 bound to antagonist strychnine. 26416729 4 30 theme X-ray 586:590 arg1 structure 592:600 a 3.0 Å X-ray structure 578:600 a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine 578:719 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 31 with complex 651:657 arg1 strychnine 710:719 strychnine 710:719 strychnine 710:719 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 31 with complex 651:657 arg1 affinity 671:678 a high affinity 664:678 a high affinity 664:678 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 31 with complex 651:657 arg1 antagonist 698:707 high-specificity antagonist 681:707 high-specificity antagonist 681:707 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 2 32 theme subsequent 416:425 arg1 opening 427:433 the subsequent opening 412:433 the subsequent opening of the ion channel 412:452 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 2 33 theme channel 446:452 arg1 insights 373:380 tremendous insights 362:380 tremendous insights into the binding of agonists 362:409 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 2 33 theme channel 446:452 arg1 mechanism 463:471 the mechanism 459:471 the mechanism of channel activation 459:493 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 2 33 theme channel 446:452 arg1 opening 427:433 the subsequent opening 412:433 the subsequent opening of the ion channel 412:452 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 0 34 theme glycine 27:33 arg1 receptor-α3 35:45 human glycine receptor-α3 21:45 human glycine receptor-α3 21:45 Crystal structure of human glycine receptor-α3 bound to antagonist strychnine. 26416729 6 35 from closure 1026:1032 arg1 domain 1071:1076 the transmembrane domain 1053:1076 the transmembrane domain 1053:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 6 36 theme transmembrane 1057:1069 arg1 domain 1071:1076 the transmembrane domain 1053:1076 the transmembrane domain 1053:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 2 37 theme ion 442:444 arg1 channel 446:452 the ion channel 438:452 the ion channel 438:452 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 6 38 from domain 1000:1005 arg1 expansion 937:945 an expansion 934:945 an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 934:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 6 39 theme receptors 913:921 arg1 inactivation 888:899 antagonist-induced inactivation 869:899 antagonist-induced inactivation of Cys-loop receptors 869:921 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 1 40 theme Cys-loop 122:129 arg1 family 140:145 the Cys-loop receptor family 118:145 the Cys-loop receptor family 118:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 2 41 theme activation 484:493 arg1 insights 373:380 tremendous insights 362:380 tremendous insights into the binding of agonists 362:409 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 2 41 theme activation 484:493 arg1 mechanism 463:471 the mechanism 459:471 the mechanism of channel activation 459:493 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 2 41 theme activation 484:493 arg1 opening 427:433 the subsequent opening 412:433 the subsequent opening of the ion channel 412:452 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 6 42 theme Cys-loop 904:911 arg1 receptors 913:921 Cys-loop receptors 904:921 Cys-loop receptors 904:921 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 1 43 theme receptor 131:138 arg1 family 140:145 the Cys-loop receptor family 118:145 the Cys-loop receptor family 118:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 6 44 theme extracellular 986:998 arg1 domain 1000:1005 the extracellular domain 982:1005 the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 982:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 1 45 theme family 140:145 arg1 channels 106:113 Neurotransmitter-gated ion channels 79:113 Neurotransmitter-gated ion channels of the Cys-loop receptor family 79:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 1 45 theme family 140:145 arg1 mediators 161:169 essential mediators 151:169 essential mediators of fast neurotransmission 151:195 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 4 46 theme homopentamer 635:646 arg1 structure 592:600 a 3.0 Å X-ray structure 578:600 a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine 578:719 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 2 47 theme X-ray 288:292 arg1 structures 294:303 Available X-ray structures 278:303 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors 278:352 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 1 48 theme many 249:252 arg1 disorders 267:275 many neurological disorders 249:275 many neurological disorders 249:275 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 6 49 from site 974:977 arg1 domain 1000:1005 the extracellular domain 982:1005 the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 982:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 26416729 4 50 theme high 666:669 arg1 strychnine 710:719 strychnine 710:719 strychnine 710:719 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 50 theme high 666:669 arg1 affinity 671:678 a high affinity 664:678 a high affinity 664:678 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 50 theme high 666:669 arg1 antagonist 698:707 high-specificity antagonist 681:707 high-specificity antagonist 681:707 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 2 51 theme Available 278:286 arg1 structures 294:303 Available X-ray structures 278:303 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors 278:352 Available X-ray structures of prokaryotic and eukaryotic Cys-loop receptors provide tremendous insights into the binding of agonists, the subsequent opening of the ion channel, and the mechanism of channel activation. 26416729 1 52 theme essential 151:159 arg1 mediators 161:169 essential mediators 151:169 essential mediators of fast neurotransmission 151:195 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 1 52 theme essential 151:159 arg1 channels 106:113 Neurotransmitter-gated ion channels 79:113 Neurotransmitter-gated ion channels of the Cys-loop receptor family 79:145 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 1 53 theme neurological 254:265 arg1 disorders 267:275 many neurological disorders 249:275 many neurological disorders 249:275 Neurotransmitter-gated ion channels of the Cys-loop receptor family are essential mediators of fast neurotransmission throughout the nervous system and are implicated in many neurological disorders. 26416729 0 54 theme antagonist 56:65 arg1 strychnine 67:76 antagonist strychnine 56:76 antagonist strychnine 56:76 Crystal structure of human glycine receptor-α3 bound to antagonist strychnine. 26416729 4 55 theme receptor-α3 623:633 arg1 homopentamer 635:646 the human glycine receptor-α3 homopentamer 605:646 the human glycine receptor-α3 homopentamer 605:646 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 4 56 from structure 592:600 arg1 complex 651:657 complex 651:657 complex with a high affinity, high-specificity antagonist, strychnine 651:719 Here we present a 3.0 Å X-ray structure of the human glycine receptor-α3 homopentamer in complex with a high affinity, high-specificity antagonist, strychnine. 26416729 6 57 from expansion 937:945 arg1 domain 1000:1005 the extracellular domain 982:1005 the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain 982:1076 Comparisons with previous structures reveal a mechanism for antagonist-induced inactivation of Cys-loop receptors, involving an expansion of the orthosteric binding site in the extracellular domain that is coupled to closure of the ion pore in the transmembrane domain. 19136951 5 0 with individuals 916:926 arg1 cblF 933:936 cblF 933:936 cblF 933:936 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 4 1 from mutations 723:731 arg1 LMBRD1 736:741 LMBRD1 736:741 LMBRD1 resulting in loss of LMBD1 function 736:777 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 2 2 theme inborn 299:304 arg1 error 306:310 the cblF inborn error 290:310 the cblF inborn error of vitamin B(12) metabolism 290:338 In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. 19136951 0 3 theme cblF 73:76 arg1 defect 78:83 the cblF defect 69:83 the cblF defect of vitamin B12 metabolism 69:109 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 3 4 theme lysosomal 605:613 arg1 protein 624:630 a lysosomal membrane protein 603:630 a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR 603:680 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 4 theme lysosomal 605:613 arg1 LMBD1 596:600 LMBD1 596:600 LMBD1 596:600 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 2 5 theme cblF 294:297 arg1 error 306:310 the cblF inborn error 290:310 the cblF inborn error of vitamin B(12) metabolism 290:338 In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. 19136951 3 6 with protein 624:630 arg1 homology 637:644 homology 637:644 homology to lipocalin membrane receptor LIMR 637:680 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 1 7 theme odd 211:213 arg1 acids 227:231 odd chain fatty acids 211:231 odd chain fatty acids 211:231 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 0 8 theme vitamin 88:94 arg1 metabolism 100:109 vitamin B12 metabolism 88:109 vitamin B12 metabolism 88:109 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 3 9 theme membrane 615:622 arg1 protein 624:630 a lysosomal membrane protein 603:630 a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR 603:680 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 9 theme membrane 615:622 arg1 LMBD1 596:600 LMBD1 596:600 LMBD1 596:600 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 4 10 theme disease 798:804 arg1 chromosomes 806:816 the 24 disease chromosomes 791:816 the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype 791:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 3 11 theme cblF 467:470 arg1 individuals 472:482 12 unrelated cblF individuals 454:482 12 unrelated cblF individuals 454:482 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 12 theme microcell-mediated 488:505 arg1 transfer 518:525 microcell-mediated chromosome transfer 488:525 microcell-mediated chromosome transfer 488:525 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 1 13 theme chain 215:219 arg1 acids 227:231 odd chain fatty acids 211:231 odd chain fatty acids 211:231 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 4 14 theme common 859:864 arg1 haplotype 874:882 a common 1.34-Mb haplotype 857:882 a common 1.34-Mb haplotype 857:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 3 15 theme lipocalin 649:657 arg1 LIMR 677:680 lipocalin membrane receptor LIMR 649:680 lipocalin membrane receptor LIMR 649:680 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 16 from mapping 443:449 arg1 individuals 472:482 12 unrelated cblF individuals 454:482 12 unrelated cblF individuals 454:482 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 1 17 theme fatty 221:225 arg1 acids 227:231 odd chain fatty acids 211:231 odd chain fatty acids 211:231 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 0 18 theme metabolism 100:109 arg1 defect 78:83 the cblF defect 69:83 the cblF defect of vitamin B12 metabolism 69:109 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 3 19 theme membrane 659:666 arg1 LIMR 677:680 lipocalin membrane receptor LIMR 649:680 lipocalin membrane receptor LIMR 649:680 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 4 20 theme frameshift 712:721 arg1 mutations 723:731 five different frameshift mutations 697:731 five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function 697:777 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 3 21 from gene 554:557 arg1 chromosome 562:571 chromosome 6q13 562:576 chromosome 6q13 562:576 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 0 22 theme B12 96:98 arg1 metabolism 100:109 vitamin B12 metabolism 88:109 vitamin B12 metabolism 88:109 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 4 23 theme function 770:777 arg1 loss 756:759 loss 756:759 loss of LMBD1 function 756:777 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 3 24 theme unrelated 457:465 arg1 individuals 472:482 12 unrelated cblF individuals 454:482 12 unrelated cblF individuals 454:482 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 25 theme chromosome 507:516 arg1 transfer 518:525 microcell-mediated chromosome transfer 488:525 microcell-mediated chromosome transfer 488:525 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 4 26 contain carrying 818:825 arg1 chromosomes 806:816 the 24 disease chromosomes 791:816 the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype 791:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 4 26 contain carrying 818:825 arg2 mutation 836:843 the same mutation 827:843 the same mutation embedded in a common 1.34-Mb haplotype 827:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 2 27 theme B 323:323 arg1 metabolism 329:338 vitamin B(12) metabolism 315:338 vitamin B(12) metabolism 315:338 In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. 19136951 1 28 theme acids 227:231 arg1 metabolism 166:175 metabolism 166:175 metabolism of branched chain amino acids and odd chain fatty acids 166:231 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 6 29 theme metabolism 1090:1099 arg1 defect 1070:1075 the cblF defect 1061:1075 the cblF defect of cobalamin metabolism 1061:1099 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 2 30 theme vitamin 315:321 arg1 metabolism 329:338 vitamin B(12) metabolism 315:338 vitamin B(12) metabolism 315:338 In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. 19136951 0 31 theme putative 20:27 arg1 exporter 49:56 a putative lysosomal cobalamin exporter 18:56 a putative lysosomal cobalamin exporter 18:56 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 6 32 theme lysosomal 1130:1138 arg1 exporter 1149:1156 a lysosomal membrane exporter 1128:1156 a lysosomal membrane exporter for cobalamin 1128:1170 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 6 32 theme lysosomal 1130:1138 arg1 LMBD1 1119:1123 LMBD1 1119:1123 LMBD1 1119:1123 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 4 33 theme embedded 845:852 arg1 mutation 836:843 the same mutation 827:843 the same mutation embedded in a common 1.34-Mb haplotype 827:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 6 34 theme cobalamin 1080:1088 arg1 metabolism 1090:1099 cobalamin metabolism 1080:1099 cobalamin metabolism 1080:1099 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 2 35 theme free 341:344 arg1 vitamin 346:352 free vitamin 341:352 free vitamin 341:352 In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. 19136951 3 36 theme receptor 668:675 arg1 LIMR 677:680 lipocalin membrane receptor LIMR 649:680 lipocalin membrane receptor LIMR 649:680 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 5 37 theme fibroblasts 901:911 arg1 Transfection 885:896 Transfection 885:896 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 885:957 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 3 38 theme homozygosity 430:441 arg1 mapping 443:449 homozygosity mapping 430:449 homozygosity mapping in 12 unrelated cblF individuals 430:482 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 0 39 theme cobalamin 39:47 arg1 exporter 49:56 a putative lysosomal cobalamin exporter 18:56 a putative lysosomal cobalamin exporter 18:56 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 6 40 theme cblF 1065:1068 arg1 defect 1070:1075 the cblF defect 1061:1075 the cblF defect of cobalamin metabolism 1061:1099 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 5 41 theme wild-type 943:951 arg1 LMBD1 953:957 wild-type LMBD1 943:957 wild-type LMBD1 943:957 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 2 42 theme metabolism 329:338 arg1 error 306:310 the cblF inborn error 290:310 the cblF inborn error of vitamin B(12) metabolism 290:338 In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. 19136951 0 43 theme lysosomal 29:37 arg1 exporter 49:56 a putative lysosomal cobalamin exporter 18:56 a putative lysosomal cobalamin exporter 18:56 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 4 44 from haplotype 874:882 arg1 embedded 845:852 embedded 845:852 embedded 845:852 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 4 45 from embedded 845:852 arg1 haplotype 874:882 a common 1.34-Mb haplotype 857:882 a common 1.34-Mb haplotype 857:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 5 46 with Transfection 885:896 arg1 LMBD1 953:957 wild-type LMBD1 943:957 wild-type LMBD1 943:957 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 4 47 theme 1.34-Mb 866:872 arg1 haplotype 874:882 a common 1.34-Mb haplotype 857:882 a common 1.34-Mb haplotype 857:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 4 48 theme LMBD1 764:768 arg1 function 770:777 LMBD1 function 764:777 LMBD1 function 764:777 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 0 49 theme exporter 49:56 arg1 Identification 0:13 Identification 0:13 Identification of a putative lysosomal cobalamin exporter 0:56 Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. 19136951 1 50 theme homocysteine 259:270 arg1 remethylation 242:254 remethylation 242:254 remethylation of homocysteine to methionine 242:284 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 4 51 theme chromosomes 806:816 arg1 chromosomes 806:816 the 24 disease chromosomes 791:816 the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype 791:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 4 51 theme chromosomes 806:816 arg1 18 785:786 18 785:786 18 785:786 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 5 52 theme coenzyme 977:984 arg1 synthesis 986:994 cobalamin coenzyme synthesis 967:994 cobalamin coenzyme synthesis 967:994 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 5 53 theme cobalamin 967:975 arg1 synthesis 986:994 cobalamin coenzyme synthesis 967:994 cobalamin coenzyme synthesis 967:994 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 1 54 from animals 154:160 arg1 essential 141:149 essential 141:149 essential 141:149 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 1 55 theme Vitamin 112:118 arg1 B 120:120 Vitamin B 112:120 Vitamin B(12) (cobalamin) 112:136 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 1 55 theme Vitamin 112:118 arg1 12 122:123 12 122:123 12 122:123 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 1 55 theme Vitamin 112:118 arg1 cobalamin 127:135 cobalamin 127:135 cobalamin 127:135 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 4 56 theme same 831:834 arg1 mutation 836:843 the same mutation 827:843 the same mutation embedded in a common 1.34-Mb haplotype 827:882 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 5 57 theme individuals 916:926 arg1 fibroblasts 901:911 fibroblasts 901:911 fibroblasts of individuals with cblF 901:936 Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. 19136951 1 58 theme branched 180:187 arg1 acids 201:205 branched chain amino acids 180:205 branched chain amino acids 180:205 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 4 59 theme different 702:710 arg1 mutations 723:731 five different frameshift mutations 697:731 five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function 697:777 We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34-Mb haplotype. 19136951 6 60 theme membrane 1140:1147 arg1 exporter 1149:1156 a lysosomal membrane exporter 1128:1156 a lysosomal membrane exporter for cobalamin 1128:1170 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 6 60 theme membrane 1140:1147 arg1 LMBD1 1119:1123 LMBD1 1119:1123 LMBD1 1119:1123 This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. 19136951 1 61 from essential 141:149 arg1 animals 154:160 animals 154:160 animals 154:160 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 1 62 theme chain 189:193 arg1 acids 201:205 branched chain amino acids 180:205 branched chain amino acids 180:205 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 3 63 theme candidate 544:552 arg1 LMBRD1 579:584 LMBRD1 579:584 LMBRD1 579:584 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 63 theme candidate 544:552 arg1 gene 554:557 a candidate gene 542:557 a candidate gene on chromosome 6q13 542:576 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 3 64 from transfer 518:525 arg1 individuals 472:482 12 unrelated cblF individuals 454:482 12 unrelated cblF individuals 454:482 Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. 19136951 1 65 theme amino 195:199 arg1 acids 201:205 branched chain amino acids 180:205 branched chain amino acids 180:205 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 19136951 1 66 theme acids 201:205 arg1 metabolism 166:175 metabolism 166:175 metabolism of branched chain amino acids and odd chain fatty acids 166:231 Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. 21047790 0 0 theme family 65:70 arg1 neurofascin 33:43 neurofascin 33:43 neurofascin 33:43 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 0 0 theme family 65:70 arg1 member 48:53 a member 46:53 a member of the L1 family of neural cell adhesion molecules 46:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 1 1 theme cell 128:131 arg1 molecules 142:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 1 2 theme neural 217:222 arg1 network 224:230 the neural network 213:230 the neural network 213:230 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 0 3 theme L1 62:63 arg1 family 65:70 the L1 family 58:70 the L1 family of neural cell adhesion molecules 58:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 4 4 with forms 649:653 arg1 patterns 680:687 independent packing patterns 660:687 independent packing patterns 660:687 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 1 5 theme adhesion 133:140 arg1 molecules 142:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 1 6 theme network 224:230 arg1 remodeling 199:208 remodeling 199:208 remodeling 199:208 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 1 6 theme network 224:230 arg1 formation 185:193 formation 185:193 formation 185:193 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 0 7 theme neural 75:80 arg1 molecules 96:104 neural cell adhesion molecules 75:104 neural cell adhesion molecules 75:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 5 8 theme large 926:930 arg1 cluster 963:969 a large but loosely packed hydrophobic cluster 924:969 a large but loosely packed hydrophobic cluster 924:969 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 6 9 theme hydrophobic 1169:1179 arg1 cluster 1181:1187 the hydrophobic cluster 1165:1187 the hydrophobic cluster 1165:1187 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 5 10 theme individual 896:905 arg1 β-sheets 911:918 two individual GFC β-sheets 892:918 two individual GFC β-sheets 892:918 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 3 11 theme several 501:507 arg1 axonin 583:588 axonin 583:588 axonin 583:588 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 11 theme several 501:507 arg1 molecules 556:564 several other immunoglobulin superfamily cell adhesion molecules 501:564 several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam 501:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 11 theme several 501:507 arg1 hemolin 574:580 hemolin 574:580 hemolin 574:580 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 11 theme several 501:507 arg1 Dscam 595:599 Dscam 595:599 Dscam 595:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 5 12 theme GFC 907:909 arg1 β-sheets 911:918 two individual GFC β-sheets 892:918 two individual GFC β-sheets 892:918 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 3 13 theme akin 493:496 arg1 shape 486:490 a horseshoe shape 474:490 a horseshoe shape 474:490 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 14 theme N-terminal 427:436 arg1 neurofascin 457:467 neurofascin 457:467 neurofascin 457:467 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 14 theme N-terminal 427:436 arg1 domains 446:452 The four N-terminal Ig-like domains 418:452 The four N-terminal Ig-like domains of neurofascin 418:467 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 0 15 theme adhesion 87:94 arg1 molecules 96:104 neural cell adhesion molecules 75:104 neural cell adhesion molecules 75:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 4 16 theme packing 672:678 arg1 patterns 680:687 independent packing patterns 660:687 independent packing patterns 660:687 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 6 17 from β-sheet 1087:1093 arg1 essential 1099:1107 essential 1099:1107 essential 1099:1107 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 17 from β-sheet 1087:1093 arg1 bonds 1059:1063 the side chain hydrogen bonds 1035:1063 the side chain hydrogen bonds at the intermolecular β-sheet 1035:1093 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 18 theme side 1039:1042 arg1 essential 1099:1107 essential 1099:1107 essential 1099:1107 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 18 theme side 1039:1042 arg1 bonds 1059:1063 the side chain hydrogen bonds 1035:1063 the side chain hydrogen bonds at the intermolecular β-sheet 1035:1093 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 3 19 theme horseshoe 476:484 arg1 shape 486:490 a horseshoe shape 474:490 a horseshoe shape 474:490 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 5 20 theme Ig-like 810:816 arg1 domain 818:823 the second Ig-like domain 799:823 the second Ig-like domain 799:823 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 2 21 theme form 359:362 arg1 structures 335:344 two crystal structures 323:344 two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member 323:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 0 22 theme cell 82:85 arg1 molecules 96:104 neural cell adhesion molecules 75:104 neural cell adhesion molecules 75:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 2 23 theme dimeric 351:357 arg1 form 359:362 a dimeric form 349:362 a dimeric form of the headpiece of neurofascin, an L1 family member 349:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 0 24 theme adhesion 11:18 arg1 mechanism 20:28 Homophilic adhesion mechanism 0:28 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules 0:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 3 25 theme other 509:513 arg1 axonin 583:588 axonin 583:588 axonin 583:588 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 25 theme other 509:513 arg1 molecules 556:564 several other immunoglobulin superfamily cell adhesion molecules 501:564 several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam 501:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 25 theme other 509:513 arg1 hemolin 574:580 hemolin 574:580 hemolin 574:580 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 25 theme other 509:513 arg1 Dscam 595:599 Dscam 595:599 Dscam 595:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 1 26 theme key 157:159 arg1 roles 161:165 key roles 157:165 key roles 157:165 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 0 27 theme Homophilic 0:9 arg1 mechanism 20:28 Homophilic adhesion mechanism 0:28 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules 0:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 6 28 theme supplementary 1194:1206 arg1 roles 1208:1212 supplementary roles 1194:1212 supplementary roles 1194:1212 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 3 29 theme immunoglobulin 515:528 arg1 axonin 583:588 axonin 583:588 axonin 583:588 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 29 theme immunoglobulin 515:528 arg1 molecules 556:564 several other immunoglobulin superfamily cell adhesion molecules 501:564 several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam 501:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 29 theme immunoglobulin 515:528 arg1 hemolin 574:580 hemolin 574:580 hemolin 574:580 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 29 theme immunoglobulin 515:528 arg1 Dscam 595:599 Dscam 595:599 Dscam 595:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 4 30 theme neurofascin 606:616 arg1 dimer 618:622 The neurofascin dimer 602:622 The neurofascin dimer 602:622 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 5 31 theme adhesion 756:763 arg1 interaction 765:775 The adhesion interaction 752:775 The adhesion interaction 752:775 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 1 32 theme homophilic 243:252 arg1 interaction 254:264 their homophilic interaction 237:264 their homophilic interaction that mediates adhesion 237:287 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 3 33 theme cell 542:545 arg1 axonin 583:588 axonin 583:588 axonin 583:588 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 33 theme cell 542:545 arg1 molecules 556:564 several other immunoglobulin superfamily cell adhesion molecules 501:564 several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam 501:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 33 theme cell 542:545 arg1 hemolin 574:580 hemolin 574:580 hemolin 574:580 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 33 theme cell 542:545 arg1 Dscam 595:599 Dscam 595:599 Dscam 595:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 5 34 theme packed 944:949 arg1 cluster 963:969 a large but loosely packed hydrophobic cluster 924:969 a large but loosely packed hydrophobic cluster 924:969 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 6 35 theme gel 998:1000 arg1 assays 1013:1018 gel filtration assays 998:1018 gel filtration assays 998:1018 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 2 36 theme neurofascin 384:394 arg1 headpiece 371:379 the headpiece 367:379 the headpiece of neurofascin, an L1 family member 367:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 0 37 theme molecules 96:104 arg1 family 65:70 the L1 family 58:70 the L1 family of neural cell adhesion molecules 58:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 3 38 theme neurofascin 457:467 arg1 neurofascin 457:467 neurofascin 457:467 neurofascin 457:467 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 38 theme neurofascin 457:467 arg1 domains 446:452 The four N-terminal Ig-like domains 418:452 The four N-terminal Ig-like domains of neurofascin 418:467 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 2 39 theme headpiece 371:379 arg1 form 359:362 a dimeric form 349:362 a dimeric form of the headpiece of neurofascin, an L1 family member 349:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 7 40 theme conserved 1239:1247 arg1 mode 1269:1272 a conserved homophilic adhesion mode 1237:1272 a conserved homophilic adhesion mode for the L1 family 1237:1290 Our structures reveal a conserved homophilic adhesion mode for the L1 family and also shed light on how the pathological mutations of L1 affect its structure and function. 21047790 3 41 theme adhesion 547:554 arg1 axonin 583:588 axonin 583:588 axonin 583:588 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 41 theme adhesion 547:554 arg1 molecules 556:564 several other immunoglobulin superfamily cell adhesion molecules 501:564 several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam 501:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 41 theme adhesion 547:554 arg1 hemolin 574:580 hemolin 574:580 hemolin 574:580 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 41 theme adhesion 547:554 arg1 Dscam 595:599 Dscam 595:599 Dscam 595:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 6 42 theme filtration 1002:1011 arg1 assays 1013:1018 gel filtration assays 998:1018 gel filtration assays 998:1018 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 7 43 theme homophilic 1249:1258 arg1 mode 1269:1272 a conserved homophilic adhesion mode 1237:1272 a conserved homophilic adhesion mode for the L1 family 1237:1290 Our structures reveal a conserved homophilic adhesion mode for the L1 family and also shed light on how the pathological mutations of L1 affect its structure and function. 21047790 0 44 theme neurofascin 33:43 arg1 mechanism 20:28 Homophilic adhesion mechanism 0:28 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules 0:104 Homophilic adhesion mechanism of neurofascin, a member of the L1 family of neural cell adhesion molecules. 21047790 5 45 theme hydrophobic 951:961 arg1 cluster 963:969 a large but loosely packed hydrophobic cluster 924:969 a large but loosely packed hydrophobic cluster 924:969 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 7 46 theme adhesion 1260:1267 arg1 mode 1269:1272 a conserved homophilic adhesion mode 1237:1272 a conserved homophilic adhesion mode for the L1 family 1237:1290 Our structures reveal a conserved homophilic adhesion mode for the L1 family and also shed light on how the pathological mutations of L1 affect its structure and function. 21047790 3 47 theme superfamily 530:540 arg1 axonin 583:588 axonin 583:588 axonin 583:588 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 47 theme superfamily 530:540 arg1 molecules 556:564 several other immunoglobulin superfamily cell adhesion molecules 501:564 several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam 501:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 47 theme superfamily 530:540 arg1 hemolin 574:580 hemolin 574:580 hemolin 574:580 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 47 theme superfamily 530:540 arg1 Dscam 595:599 Dscam 595:599 Dscam 595:599 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 6 48 theme hydrogen 1050:1057 arg1 essential 1099:1107 essential 1099:1107 essential 1099:1107 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 48 theme hydrogen 1050:1057 arg1 bonds 1059:1063 the side chain hydrogen bonds 1035:1063 the side chain hydrogen bonds at the intermolecular β-sheet 1035:1093 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 3 49 theme Ig-like 438:444 arg1 neurofascin 457:467 neurofascin 457:467 neurofascin 457:467 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 3 49 theme Ig-like 438:444 arg1 domains 446:452 The four N-terminal Ig-like domains 418:452 The four N-terminal Ig-like domains of neurofascin 418:467 The four N-terminal Ig-like domains of neurofascin form a horseshoe shape, akin to several other immunoglobulin superfamily cell adhesion molecules such as hemolin, axonin, and Dscam. 21047790 2 50 theme family 403:408 arg1 member 410:415 an L1 family member 397:415 an L1 family member 397:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 2 50 theme family 403:408 arg1 neurofascin 384:394 neurofascin 384:394 neurofascin 384:394 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 7 51 theme L1 1282:1283 arg1 family 1285:1290 the L1 family 1278:1290 the L1 family 1278:1290 Our structures reveal a conserved homophilic adhesion mode for the L1 family and also shed light on how the pathological mutations of L1 affect its structure and function. 21047790 7 52 theme pathological 1323:1334 arg1 mutations 1336:1344 the pathological mutations 1319:1344 the pathological mutations of L1 1319:1350 Our structures reveal a conserved homophilic adhesion mode for the L1 family and also shed light on how the pathological mutations of L1 affect its structure and function. 21047790 1 53 theme neural 121:126 arg1 molecules 142:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 4 54 theme crystal 641:647 arg1 forms 649:653 two crystal forms 637:653 two crystal forms with independent packing patterns 637:687 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 2 55 theme crystal 327:333 arg1 structures 335:344 two crystal structures 323:344 two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member 323:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 6 56 theme homophilic 1117:1126 arg1 interaction 1128:1138 the homophilic interaction 1113:1138 the homophilic interaction 1113:1138 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 4 57 theme trans-synaptic 722:735 arg1 mode 746:749 trans-synaptic adhesion mode 722:749 trans-synaptic adhesion mode 722:749 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 4 58 theme independent 660:670 arg1 patterns 680:687 independent packing patterns 660:687 independent packing patterns 660:687 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 7 59 theme L1 1349:1350 arg1 mutations 1336:1344 the pathological mutations 1319:1344 the pathological mutations of L1 1319:1350 Our structures reveal a conserved homophilic adhesion mode for the L1 family and also shed light on how the pathological mutations of L1 affect its structure and function. 21047790 4 60 theme horseshoes 708:717 arg1 pair 700:703 a pair 698:703 a pair of horseshoes 698:717 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 5 61 theme intermolecular 844:857 arg1 β-sheet 859:865 an intermolecular β-sheet 841:865 an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster 841:969 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 1 62 dep formation 185:193 arg1 the 181:183 the 181:183 the 181:183 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 1 63 theme L1 111:112 arg1 molecules 142:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 6 64 theme intermolecular 1072:1085 arg1 β-sheet 1087:1093 the intermolecular β-sheet 1068:1093 the intermolecular β-sheet 1068:1093 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 65 from cluster 1181:1187 arg1 residues 1153:1160 the residues 1149:1160 the residues at the hydrophobic cluster 1149:1187 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 66 theme chain 1044:1048 arg1 essential 1099:1107 essential 1099:1107 essential 1099:1107 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 6 66 theme chain 1044:1048 arg1 bonds 1059:1063 the side chain hydrogen bonds 1035:1063 the side chain hydrogen bonds at the intermolecular β-sheet 1035:1093 Mutagenesis combined with gel filtration assays suggested that the side chain hydrogen bonds at the intermolecular β-sheet are essential for the homophilic interaction and that the residues at the hydrophobic cluster play supplementary roles. 21047790 1 67 theme family 114:119 arg1 molecules 142:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules 107:150 The L1 family neural cell adhesion molecules play key roles in specifying the formation and remodeling of the neural network, but their homophilic interaction that mediates adhesion is not well understood. 21047790 5 68 theme second 803:808 arg1 domain 818:823 the second Ig-like domain 799:823 the second Ig-like domain 799:823 The adhesion interaction is mediated mostly by the second Ig-like domain, which features an intermolecular β-sheet formed by the joining of two individual GFC β-sheets and a large but loosely packed hydrophobic cluster. 21047790 4 69 theme adhesion 737:744 arg1 mode 746:749 trans-synaptic adhesion mode 722:749 trans-synaptic adhesion mode 722:749 The neurofascin dimer, captured in two crystal forms with independent packing patterns, reveals a pair of horseshoes in trans-synaptic adhesion mode. 21047790 2 70 theme L1 400:401 arg1 member 410:415 an L1 family member 397:415 an L1 family member 397:415 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 21047790 2 70 theme L1 400:401 arg1 neurofascin 384:394 neurofascin 384:394 neurofascin 384:394 We report two crystal structures of a dimeric form of the headpiece of neurofascin, an L1 family member. 26235030 4 0 from UNC5 613:616 arg1 distinct 599:606 distinct 599:606 distinct 599:606 The LPHN3/FLRT3 structure reveals that LPHN3 binds to FLRT3 at a site distinct from UNC5. 26235030 8 1 theme synapse 1151:1157 arg1 function 1159:1166 synapse function 1151:1166 synapse function 1151:1166 These findings provide molecular insights for understanding the role of cell-adhesion proteins in synapse function. 26235030 2 2 theme the developing 417:430 arg1 cortex 432:437 the developing cortex 417:437 the developing cortex 417:437 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 0 3 from Basis 11:15 arg1 Adhesion 62:69 Cell Adhesion 57:69 Cell Adhesion 57:69 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 26235030 1 4 from functions 177:185 arg1 development 199:209 cortical development 190:209 cortical development 190:209 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 4 from functions 177:185 arg1 formation 223:231 synapse formation 215:231 synapse formation 215:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 7 5 theme transsynaptic 1003:1015 arg1 complexes 1017:1025 transsynaptic complexes 1003:1025 transsynaptic complexes with both LPHN3 and UNC5 1003:1050 Our results suggest that UNC5 and LPHN3 can simultaneously bind to FLRT3, forming a trimeric complex, and that FLRT3 may form transsynaptic complexes with both LPHN3 and UNC5. 26235030 4 6 theme distinct 599:606 arg1 site 594:597 a site 592:597 a site distinct from UNC5 592:616 The LPHN3/FLRT3 structure reveals that LPHN3 binds to FLRT3 at a site distinct from UNC5. 26235030 5 7 theme cell-membrane 754:766 arg1 localization 768:779 their cell-membrane localization 748:779 their cell-membrane localization 748:779 Structure-based mutations specifically disrupt LPHN3/FLRT3 binding, but do not disturb their interactions with other proteins or their cell-membrane localization. 26235030 5 8 theme other 730:734 arg1 proteins 736:743 other proteins 730:743 other proteins 730:743 Structure-based mutations specifically disrupt LPHN3/FLRT3 binding, but do not disturb their interactions with other proteins or their cell-membrane localization. 26235030 1 9 theme Fibronectin 72:82 arg1 repeat 97:102 Fibronectin leucine-rich repeat 72:102 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) 72:133 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 7 10 with complexes 1017:1025 arg1 LPHN3 1037:1041 LPHN3 1037:1041 LPHN3 1037:1041 Our results suggest that UNC5 and LPHN3 can simultaneously bind to FLRT3, forming a trimeric complex, and that FLRT3 may form transsynaptic complexes with both LPHN3 and UNC5. 26235030 7 10 with complexes 1017:1025 arg1 UNC5 1047:1050 UNC5 1047:1050 UNC5 1047:1050 Our results suggest that UNC5 and LPHN3 can simultaneously bind to FLRT3, forming a trimeric complex, and that FLRT3 may form transsynaptic complexes with both LPHN3 and UNC5. 26235030 5 11 theme Structure-based 619:633 arg1 mutations 635:643 Structure-based mutations 619:643 Structure-based mutations 619:643 Structure-based mutations specifically disrupt LPHN3/FLRT3 binding, but do not disturb their interactions with other proteins or their cell-membrane localization. 26235030 1 12 theme leucine-rich 84:95 arg1 repeat 97:102 Fibronectin leucine-rich repeat 72:102 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) 72:133 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 13 theme cortical 190:197 arg1 development 199:209 cortical development 190:209 cortical development 190:209 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 6 14 used used 800:803 arg2 they 788:791 they 788:791 they 788:791 Thus, they can be used as molecular tools to dissect the functions of FLRTs and LPHNs in vivo. 26235030 6 14 used used 800:803 arg2 tools 818:822 molecular tools 808:822 molecular tools 808:822 Thus, they can be used as molecular tools to dissect the functions of FLRTs and LPHNs in vivo. 26235030 1 15 with molecules 153:161 arg1 functions 177:185 emerging functions 168:185 emerging functions in cortical development and synapse formation 168:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 6 16 theme molecular 808:816 arg1 they 788:791 they 788:791 they 788:791 Thus, they can be used as molecular tools to dissect the functions of FLRTs and LPHNs in vivo. 26235030 6 16 theme molecular 808:816 arg1 tools 818:822 molecular tools 808:822 molecular tools 808:822 Thus, they can be used as molecular tools to dissect the functions of FLRTs and LPHNs in vivo. 26235030 1 17 theme repeat 97:102 arg1 proteins 118:125 Fibronectin leucine-rich repeat transmembrane proteins 72:125 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) 72:133 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 17 theme repeat 97:102 arg1 molecules 153:161 cell-adhesion molecules 139:161 cell-adhesion molecules with emerging functions in cortical development and synapse formation 139:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 17 theme repeat 97:102 arg1 FLRTs 128:132 FLRTs 128:132 FLRTs 128:132 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 3 18 theme crystal 461:467 arg1 structures 469:478 the crystal structures 457:478 the crystal structures of FLRT3 in isolation and in complex with LPHN3 457:526 Here, we present the crystal structures of FLRT3 in isolation and in complex with LPHN3. 26235030 1 19 theme transmembrane 104:116 arg1 proteins 118:125 Fibronectin leucine-rich repeat transmembrane proteins 72:125 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) 72:133 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 19 theme transmembrane 104:116 arg1 molecules 153:161 cell-adhesion molecules 139:161 cell-adhesion molecules with emerging functions in cortical development and synapse formation 139:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 19 theme transmembrane 104:116 arg1 FLRTs 128:132 FLRTs 128:132 FLRTs 128:132 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 2 20 theme neurons 406:412 arg1 migration 393:401 the migration 389:401 the migration of neurons in the developing cortex 389:437 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 0 21 theme Structural 0:9 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 0:70 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 26235030 3 22 with complex 509:515 arg1 LPHN3 522:526 LPHN3 522:526 LPHN3 522:526 Here, we present the crystal structures of FLRT3 in isolation and in complex with LPHN3. 26235030 2 23 from migration 393:401 arg1 cortex 432:437 the developing cortex 417:437 the developing cortex 417:437 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 2 24 theme Uncoordinated-5 338:352 arg1 receptors 368:376 Uncoordinated-5 (UNC5)/netrin receptors 338:376 Uncoordinated-5 (UNC5)/netrin receptors 338:376 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 1 25 theme synapse 215:221 arg1 formation 223:231 synapse formation 215:231 synapse formation 215:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 0 26 theme Latrophilin-FLRT-UNC5 20:40 arg1 Interaction 42:52 Latrophilin-FLRT-UNC5 Interaction 20:52 Latrophilin-FLRT-UNC5 Interaction 20:52 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 26235030 8 27 theme molecular 1076:1084 arg1 insights 1086:1093 molecular insights 1076:1093 molecular insights for understanding the role of cell-adhesion proteins in synapse function 1076:1166 These findings provide molecular insights for understanding the role of cell-adhesion proteins in synapse function. 26235030 7 28 theme trimeric 961:968 arg1 complex 970:976 a trimeric complex 959:976 a trimeric complex 959:976 Our results suggest that UNC5 and LPHN3 can simultaneously bind to FLRT3, forming a trimeric complex, and that FLRT3 may form transsynaptic complexes with both LPHN3 and UNC5. 26235030 3 29 theme FLRT3 483:487 arg1 structures 469:478 the crystal structures 457:478 the crystal structures of FLRT3 in isolation and in complex with LPHN3 457:526 Here, we present the crystal structures of FLRT3 in isolation and in complex with LPHN3. 26235030 2 30 theme synapse 308:314 arg1 development 316:326 synapse development 308:326 synapse development 308:326 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 0 31 theme Interaction 42:52 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 0:70 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 26235030 8 32 from role 1117:1120 arg1 function 1159:1166 synapse function 1151:1166 synapse function 1151:1166 These findings provide molecular insights for understanding the role of cell-adhesion proteins in synapse function. 26235030 2 33 from neurons 406:412 arg1 cortex 432:437 the developing cortex 417:437 the developing cortex 417:437 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 4 34 theme LPHN3/FLRT3 533:543 arg1 structure 545:553 The LPHN3/FLRT3 structure 529:553 The LPHN3/FLRT3 structure 529:553 The LPHN3/FLRT3 structure reveals that LPHN3 binds to FLRT3 at a site distinct from UNC5. 26235030 6 35 theme LPHNs 862:866 arg1 functions 839:847 the functions 835:847 the functions of FLRTs and LPHNs in vivo 835:874 Thus, they can be used as molecular tools to dissect the functions of FLRTs and LPHNs in vivo. 26235030 0 36 theme Cell 57:60 arg1 Adhesion 62:69 Cell Adhesion 57:69 Cell Adhesion 57:69 Structural Basis of Latrophilin-FLRT-UNC5 Interaction in Cell Adhesion. 26235030 2 37 theme extracellular 240:252 arg1 regions 254:260 Their extracellular regions 234:260 Their extracellular regions 234:260 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 2 38 theme /netrin 360:366 arg1 receptors 368:376 Uncoordinated-5 (UNC5)/netrin receptors 338:376 Uncoordinated-5 (UNC5)/netrin receptors 338:376 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 6 39 theme FLRTs 852:856 arg1 functions 839:847 the functions 835:847 the functions of FLRTs and LPHNs in vivo 835:874 Thus, they can be used as molecular tools to dissect the functions of FLRTs and LPHNs in vivo. 26235030 1 40 theme cell-adhesion 139:151 arg1 proteins 118:125 Fibronectin leucine-rich repeat transmembrane proteins 72:125 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) 72:133 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 1 40 theme cell-adhesion 139:151 arg1 molecules 153:161 cell-adhesion molecules 139:161 cell-adhesion molecules with emerging functions in cortical development and synapse formation 139:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 3 41 from structures 469:478 arg1 isolation 492:500 isolation 492:500 isolation 492:500 Here, we present the crystal structures of FLRT3 in isolation and in complex with LPHN3. 26235030 3 41 from structures 469:478 arg1 complex 509:515 complex 509:515 complex with LPHN3 509:526 Here, we present the crystal structures of FLRT3 in isolation and in complex with LPHN3. 26235030 2 42 theme UNC5 355:358 arg1 receptors 368:376 Uncoordinated-5 (UNC5)/netrin receptors 338:376 Uncoordinated-5 (UNC5)/netrin receptors 338:376 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 8 43 theme cell-adhesion 1125:1137 arg1 proteins 1139:1146 cell-adhesion proteins 1125:1146 cell-adhesion proteins 1125:1146 These findings provide molecular insights for understanding the role of cell-adhesion proteins in synapse function. 26235030 8 44 theme proteins 1139:1146 arg1 role 1117:1120 the role 1113:1120 the role of cell-adhesion proteins in synapse function 1113:1166 These findings provide molecular insights for understanding the role of cell-adhesion proteins in synapse function. 26235030 2 45 from cortex 432:437 arg1 migration 393:401 the migration 389:401 the migration of neurons in the developing cortex 389:437 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 2 46 dep mediate 300:306 arg1 control 381:387 control 381:387 to control the migration of neurons in the developing cortex 378:437 Their extracellular regions interact with latrophilins (LPHNs) to mediate synapse development, and with Uncoordinated-5 (UNC5)/netrin receptors to control the migration of neurons in the developing cortex. 26235030 5 47 with interactions 712:723 arg1 localization 768:779 their cell-membrane localization 748:779 their cell-membrane localization 748:779 Structure-based mutations specifically disrupt LPHN3/FLRT3 binding, but do not disturb their interactions with other proteins or their cell-membrane localization. 26235030 5 47 with interactions 712:723 arg1 proteins 736:743 other proteins 730:743 other proteins 730:743 Structure-based mutations specifically disrupt LPHN3/FLRT3 binding, but do not disturb their interactions with other proteins or their cell-membrane localization. 26235030 1 48 theme emerging 168:175 arg1 functions 177:185 emerging functions 168:185 emerging functions in cortical development and synapse formation 168:231 Fibronectin leucine-rich repeat transmembrane proteins (FLRTs) are cell-adhesion molecules with emerging functions in cortical development and synapse formation. 26235030 5 49 theme LPHN3/FLRT3 666:676 arg1 binding 678:684 LPHN3/FLRT3 binding 666:684 LPHN3/FLRT3 binding 666:684 Structure-based mutations specifically disrupt LPHN3/FLRT3 binding, but do not disturb their interactions with other proteins or their cell-membrane localization. 3353370 1 0 theme platelet 216:223 arg1 receptor 225:232 the platelet receptor 212:232 the platelet receptor for von Willebrand factor 212:258 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 0 theme platelet 216:223 arg1 Ib 201:202 human glycoprotein Ib 182:202 human glycoprotein Ib (GPIb) 182:209 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 10 1 theme protein 2085:2091 arg1 kinase 2093:2098 cAMP-dependent protein kinase 2070:2098 cAMP-dependent protein kinase 2070:2098 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 4 2 theme nucleotides 794:804 arg1 frame 781:785 an open reading frame 765:785 an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon 765:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 2 theme nucleotides 794:804 arg1 region 739:744 a 5' untranslated region 721:744 a 5' untranslated region of 22 nucleotides 721:762 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 2 theme nucleotides 794:804 arg1 region 924:929 a 3' noncoding region 909:929 a 3' noncoding region of 307 nucleotides 909:948 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 7 3 theme amino-terminal 1290:1303 arg1 region 1305:1310 The amino-terminal region 1286:1310 The amino-terminal region of the beta chain 1286:1328 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 5 4 from tail 1069:1072 arg1 upstream 1043:1050 upstream 1043:1050 upstream 1043:1050 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 7 5 theme chain 1324:1328 arg1 region 1305:1310 The amino-terminal region 1286:1310 The amino-terminal region of the beta chain 1286:1328 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 6 6 theme intact 1118:1123 arg1 chain 1130:1134 the intact beta chain 1114:1134 the intact beta chain 1114:1134 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 8 7 theme GPIb 1583:1586 arg1 chain 1574:1578 the beta chain 1565:1578 the beta chain of GPIb 1565:1586 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 1 8 theme chain 173:177 arg1 structure 151:159 The primary structure 139:159 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, 139:259 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 10 9 theme cAMP-dependent 2070:2083 arg1 kinase 2093:2098 cAMP-dependent protein kinase 2070:2098 cAMP-dependent protein kinase 2070:2098 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 9 10 theme amino-terminal 1774:1787 arg1 region 1789:1794 The amino-terminal region 1770:1794 The amino-terminal region of the beta chain of GPIb 1770:1820 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 10 11 contain contains 1993:2000 arg2 cysteine 2014:2021 an unpaired cysteine 2002:2021 an unpaired cysteine 2002:2021 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 10 11 contain contains 1993:2000 arg1 segment 1985:1991 The intracellular segment 1967:1991 The intracellular segment 1967:1991 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 10 11 contain contains 1993:2000 arg2 sites 2041:2045 two potential sites 2027:2045 two potential sites for phosphorylation by cAMP-dependent protein kinase 2027:2098 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 0 12 theme leucine-rich 105:116 arg1 sequence 129:136 a leucine-rich amino acid sequence 103:136 a leucine-rich amino acid sequence 103:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 5 13 theme noncoding 958:966 arg1 sequence 968:975 The 3' noncoding sequence 951:975 The 3' noncoding sequence 951:975 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 1 14 theme human 182:186 arg1 receptor 225:232 the platelet receptor 212:232 the platelet receptor for von Willebrand factor 212:258 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 14 theme human 182:186 arg1 GPIb 205:208 GPIb 205:208 GPIb 205:208 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 14 theme human 182:186 arg1 Ib 201:202 human glycoprotein Ib 182:202 human glycoprotein Ib (GPIb) 182:209 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 2 15 theme polyclonal 500:509 arg1 antibody 511:518 a radiolabeled affinity-purified rabbit polyclonal antibody 460:518 a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb 460:544 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 0 16 theme acid 124:127 arg1 sequence 129:136 a leucine-rich amino acid sequence 103:136 a leucine-rich amino acid sequence 103:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 3 17 theme positive 556:563 arg1 clones 565:570 Eighteen positive clones 547:570 Eighteen positive clones 547:570 Eighteen positive clones were isolated and plaque-purified and the nucleotide sequences of three were determined. 3353370 1 18 theme Ib 201:202 arg1 chain 173:177 the beta chain 164:177 the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, 164:259 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 7 19 theme GPIb 1469:1472 arg1 chain 1460:1464 the alpha chain 1450:1464 the alpha chain of GPIb 1450:1472 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 4 20 theme acids 844:848 arg1 peptide 824:830 a signal peptide 815:830 a signal peptide of 28 amino acids 815:848 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 20 theme acids 844:848 arg1 codon 898:902 a stop codon 891:902 a stop codon 891:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 20 theme acids 844:848 arg1 protein 863:869 a mature protein 854:869 a mature protein of 181 amino acids 854:888 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 1 21 theme Willebrand 242:251 arg1 factor 253:258 von Willebrand factor 238:258 von Willebrand factor 238:258 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 9 22 theme beta 1803:1806 arg1 chain 1808:1812 the beta chain 1799:1812 the beta chain of GPIb 1799:1820 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 8 23 theme alpha 1711:1715 arg1 chain 1717:1721 the alpha chain 1707:1721 the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1707:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 2 24 theme beta 527:530 arg1 chain 532:536 the beta chain 523:536 the beta chain of GPIb 523:544 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 10 25 theme intracellular 1971:1983 arg1 segment 1985:1991 The intracellular segment 1967:1991 The intracellular segment 1967:1991 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 6 26 theme chemical 1164:1171 arg1 cleavage 1173:1180 chemical cleavage 1164:1180 chemical cleavage 1164:1180 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 8 27 theme acid 1622:1625 arg1 sequences 1627:1635 amino acid sequences 1616:1635 amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1616:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 7 28 contain contains 1330:1337 arg1 region 1305:1310 The amino-terminal region 1286:1310 The amino-terminal region of the beta chain 1286:1328 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 28 contain contains 1330:1337 arg2 sequence 1354:1361 a leucine-rich sequence 1339:1361 a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein 1339:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 28 contain contains 1330:1337 arg2 similar 1389:1395 similar 1389:1395 similar 1389:1395 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 0 29 theme glycoprotein 44:55 arg1 Ib 57:58 human platelet glycoprotein Ib 29:58 human platelet glycoprotein Ib 29:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 5 30 theme AATAAA 1020:1025 arg1 nucleotides 1031:1041 a polyadenylylation signal (AATAAA) 14 nucleotides 992:1041 a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides 992:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 10 31 theme unpaired 2005:2012 arg1 cysteine 2014:2021 an unpaired cysteine 2002:2021 an unpaired cysteine 2002:2021 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 4 32 theme composite 665:673 arg1 sequence 675:682 The composite sequence 661:682 The composite sequence 661:682 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 2 33 theme affinity-purified 475:491 arg1 antibody 511:518 a radiolabeled affinity-purified rabbit polyclonal antibody 460:518 a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb 460:544 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 9 34 theme transmembrane 1839:1851 arg1 segment 1853:1859 a transmembrane segment 1837:1859 a transmembrane segment of 25 amino acids 1837:1877 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 4 35 theme untranslated 726:737 arg1 region 739:744 a 5' untranslated region 721:744 a 5' untranslated region of 22 nucleotides 721:762 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 0 36 theme transmembrane 69:81 arg1 proteins 83:90 both transmembrane proteins 64:90 both transmembrane proteins containing a leucine-rich amino acid sequence 64:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 0 36 theme transmembrane 69:81 arg1 chains 19:24 The alpha and beta chains 0:24 The alpha and beta chains of human platelet glycoprotein Ib 0:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 1 37 theme cloning 307:313 arg1 combination 287:297 a combination 285:297 a combination of cDNA cloning and amino acid sequence analysis 285:346 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 10 38 theme potential 2031:2039 arg1 sites 2041:2045 two potential sites 2027:2045 two potential sites for phosphorylation by cAMP-dependent protein kinase 2027:2098 The intracellular segment contains an unpaired cysteine and two potential sites for phosphorylation by cAMP-dependent protein kinase. 3353370 1 39 theme amino 319:323 arg1 sequence 330:337 amino acid sequence 319:337 amino acid sequence analysis 319:346 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 5 40 contain contained 982:990 arg2 nucleotides 1031:1041 a polyadenylylation signal (AATAAA) 14 nucleotides 992:1041 a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides 992:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 5 40 contain contained 982:990 arg1 sequence 968:975 The 3' noncoding sequence 951:975 The 3' noncoding sequence 951:975 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 9 41 theme amino 1867:1871 arg1 acids 1873:1877 25 amino acids 1864:1877 25 amino acids 1864:1877 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 8 42 from sequence 1553:1560 arg1 chain 1574:1578 the beta chain 1565:1578 the beta chain of GPIb 1565:1586 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 8 43 theme alpha 1748:1752 arg1 2-glycoprotein 1754:1767 leucine-rich alpha 2-glycoprotein 1735:1767 leucine-rich alpha 2-glycoprotein 1735:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 1 44 theme von 238:240 arg1 factor 253:258 von Willebrand factor 238:258 von Willebrand factor 238:258 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 7 45 from repeats 1490:1496 arg1 2-glycoprotein 1520:1533 leucine-rich alpha 2-glycoprotein 1501:1533 leucine-rich alpha 2-glycoprotein 1501:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 45 from repeats 1490:1496 arg1 chain 1460:1464 the alpha chain 1450:1464 the alpha chain of GPIb 1450:1472 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 8 46 theme tandem 1689:1694 arg1 repeats 1696:1702 the leucine-rich tandem repeats 1672:1702 the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1672:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 7 47 theme alpha 1514:1518 arg1 2-glycoprotein 1520:1533 leucine-rich alpha 2-glycoprotein 1501:1533 leucine-rich alpha 2-glycoprotein 1501:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 9 48 theme intracellular 1886:1898 arg1 segment 1900:1906 an intracellular segment 1883:1906 an intracellular segment of 34 amino acids 1883:1924 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 4 49 theme nucleotides 938:948 arg1 frame 781:785 an open reading frame 765:785 an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon 765:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 49 theme nucleotides 938:948 arg1 region 739:744 a 5' untranslated region 721:744 a 5' untranslated region of 22 nucleotides 721:762 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 49 theme nucleotides 938:948 arg1 region 924:929 a 3' noncoding region 909:929 a 3' noncoding region of 307 nucleotides 909:948 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 2 50 theme expression 369:378 arg1 library 380:386 A lambda phage cDNA expression library 349:386 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) 349:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 2 51 theme HEL 431:433 arg1 cells 424:428 human erythroleukemia cells 402:428 human erythroleukemia cells (HEL cells) 402:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 2 51 theme HEL 431:433 arg1 cells 435:439 HEL cells 431:439 HEL cells 431:439 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 3 52 theme nucleotide 614:623 arg1 sequences 625:633 the nucleotide sequences 610:633 the nucleotide sequences of three 610:642 Eighteen positive clones were isolated and plaque-purified and the nucleotide sequences of three were determined. 3353370 4 53 theme reading 773:779 arg1 frame 781:785 an open reading frame 765:785 an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon 765:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 2 54 theme phage 358:362 arg1 library 380:386 A lambda phage cDNA expression library 349:386 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) 349:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 5 55 theme upstream 1043:1050 arg1 nucleotides 1031:1041 a polyadenylylation signal (AATAAA) 14 nucleotides 992:1041 a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides 992:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 4 56 theme noncoding 914:922 arg1 region 924:929 a 3' noncoding region 909:929 a 3' noncoding region of 307 nucleotides 909:948 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 0 57 theme human 29:33 arg1 Ib 57:58 human platelet glycoprotein Ib 29:58 human platelet glycoprotein Ib 29:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 7 58 from repeats 1439:1445 arg1 2-glycoprotein 1520:1533 leucine-rich alpha 2-glycoprotein 1501:1533 leucine-rich alpha 2-glycoprotein 1501:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 58 from repeats 1439:1445 arg1 chain 1460:1464 the alpha chain 1450:1464 the alpha chain of GPIb 1450:1472 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 6 59 theme chain 1130:1134 arg1 degradation 1099:1109 Edman degradation 1093:1109 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage 1093:1180 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 9 60 theme amino 1914:1918 arg1 acids 1920:1924 34 amino acids 1911:1924 34 amino acids 1911:1924 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 4 61 theme nucleotides 752:762 arg1 frame 781:785 an open reading frame 765:785 an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon 765:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 61 theme nucleotides 752:762 arg1 region 739:744 a 5' untranslated region 721:744 a 5' untranslated region of 22 nucleotides 721:762 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 61 theme nucleotides 752:762 arg1 region 924:929 a 3' noncoding region 909:929 a 3' noncoding region of 307 nucleotides 909:948 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 7 62 theme amino 1369:1373 arg1 acids 1375:1379 24 amino acids 1366:1379 24 amino acids 1366:1379 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 63 gly 2-glycoprotein 1520:1533 arg1 2-glycoprotein 1520:1533 leucine-rich alpha 2-glycoprotein 1501:1533 leucine-rich alpha 2-glycoprotein 1501:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 8 64 theme chain 1717:1721 arg1 repeats 1696:1702 the leucine-rich tandem repeats 1672:1702 the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1672:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 9 65 theme carboxyl 1933:1940 arg1 terminus 1942:1949 the carboxyl terminus 1929:1949 the carboxyl terminus of the protein 1929:1964 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 7 66 theme leucine-rich 1341:1352 arg1 sequence 1354:1361 a leucine-rich sequence 1339:1361 a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein 1339:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 66 theme leucine-rich 1341:1352 arg1 similar 1389:1395 similar 1389:1395 similar 1389:1395 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 4 67 theme signal 817:822 arg1 peptide 824:830 a signal peptide 815:830 a signal peptide of 28 amino acids 815:848 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 6 68 theme peptides 1143:1150 arg1 degradation 1099:1109 Edman degradation 1093:1109 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage 1093:1180 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 8 69 theme leucine-rich 1540:1551 arg1 sequence 1553:1560 The leucine-rich sequence 1536:1560 The leucine-rich sequence in the beta chain of GPIb 1536:1586 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 5 70 theme nucleotides 1080:1090 arg1 tail 1069:1072 the poly(A) tail 1057:1072 the poly(A) tail of 18 nucleotides 1057:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 2 71 theme GPIb 541:544 arg1 chain 532:536 the beta chain 523:536 the beta chain of GPIb 523:544 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 9 72 theme protein 1958:1964 arg1 terminus 1942:1949 the carboxyl terminus 1929:1949 the carboxyl terminus of the protein 1929:1964 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 7 73 theme beta 1319:1322 arg1 chain 1324:1328 the beta chain 1315:1328 the beta chain 1315:1328 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 8 74 gly 2-glycoprotein 1754:1767 arg1 2-glycoprotein 1754:1767 leucine-rich alpha 2-glycoprotein 1735:1767 leucine-rich alpha 2-glycoprotein 1735:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 6 75 theme beta 1125:1128 arg1 chain 1130:1134 the intact beta chain 1114:1134 the intact beta chain 1114:1134 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 8 76 theme beta 1569:1572 arg1 chain 1574:1578 the beta chain 1565:1578 the beta chain of GPIb 1565:1586 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 1 77 theme beta 168:171 arg1 chain 173:177 the beta chain 164:177 the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, 164:259 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 8 78 theme GPIb 1726:1729 arg1 chain 1717:1721 the alpha chain 1707:1721 the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1707:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 7 79 theme tandem 1483:1488 arg1 repeats 1490:1496 nine tandem repeats 1478:1496 nine tandem repeats in leucine-rich alpha 2-glycoprotein 1478:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 1 80 gly glycoprotein 188:199 arg1 glycoprotein 188:199 human glycoprotein Ib 182:202 human glycoprotein Ib (GPIb) 182:209 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 0 81 theme amino 118:122 arg1 sequence 129:136 a leucine-rich amino acid sequence 103:136 a leucine-rich amino acid sequence 103:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 4 82 theme mature 856:861 arg1 protein 863:869 a mature protein 854:869 a mature protein of 181 amino acids 854:888 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 82 theme mature 856:861 arg1 codon 898:902 a stop codon 891:902 a stop codon 891:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 1 83 theme analysis 339:346 arg1 combination 287:297 a combination 285:297 a combination of cDNA cloning and amino acid sequence analysis 285:346 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 84 theme glycoprotein 188:199 arg1 receptor 225:232 the platelet receptor 212:232 the platelet receptor for von Willebrand factor 212:258 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 84 theme glycoprotein 188:199 arg1 GPIb 205:208 GPIb 205:208 GPIb 205:208 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 84 theme glycoprotein 188:199 arg1 Ib 201:202 human glycoprotein Ib 182:202 human glycoprotein Ib (GPIb) 182:209 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 2 85 theme rabbit 493:498 arg1 antibody 511:518 a radiolabeled affinity-purified rabbit polyclonal antibody 460:518 a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb 460:544 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 4 86 theme acids 884:888 arg1 peptide 824:830 a signal peptide 815:830 a signal peptide of 28 amino acids 815:848 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 86 theme acids 884:888 arg1 codon 898:902 a stop codon 891:902 a stop codon 891:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 86 theme acids 884:888 arg1 protein 863:869 a mature protein 854:869 a mature protein of 181 amino acids 854:888 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 6 87 theme amino 1190:1194 arg1 sequences 1201:1209 amino acid sequences 1190:1209 amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA 1190:1283 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 7 88 theme alpha 1454:1458 arg1 chain 1460:1464 the alpha chain 1450:1464 the alpha chain of GPIb 1450:1472 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 4 89 theme amino 838:842 arg1 acids 844:848 28 amino acids 835:848 28 amino acids 835:848 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 6 90 theme Edman 1093:1097 arg1 degradation 1099:1109 Edman degradation 1093:1109 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage 1093:1180 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 9 91 theme chain 1808:1812 arg1 region 1789:1794 The amino-terminal region 1770:1794 The amino-terminal region of the beta chain of GPIb 1770:1820 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 9 92 theme GPIb 1817:1820 arg1 chain 1808:1812 the beta chain 1799:1812 the beta chain of GPIb 1799:1820 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 0 93 theme Ib 57:58 arg1 proteins 83:90 both transmembrane proteins 64:90 both transmembrane proteins containing a leucine-rich amino acid sequence 64:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 0 93 theme Ib 57:58 arg1 chains 19:24 The alpha and beta chains 0:24 The alpha and beta chains of human platelet glycoprotein Ib 0:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 8 94 theme amino 1616:1620 arg1 sequences 1627:1635 amino acid sequences 1616:1635 amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1616:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 7 95 theme tandem 1432:1437 arg1 repeats 1439:1445 seven tandem repeats 1426:1445 seven tandem repeats in the alpha chain of GPIb 1426:1472 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 4 96 theme stop 893:896 arg1 codon 898:902 a stop codon 891:902 a stop codon 891:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 4 96 theme stop 893:896 arg1 protein 863:869 a mature protein 854:869 a mature protein of 181 amino acids 854:888 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 2 97 theme radiolabeled 462:473 arg1 antibody 511:518 a radiolabeled affinity-purified rabbit polyclonal antibody 460:518 a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb 460:544 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 1 98 theme primary 143:149 arg1 structure 151:159 The primary structure 139:159 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, 139:259 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 8 99 theme leucine-rich 1735:1746 arg1 2-glycoprotein 1754:1767 leucine-rich alpha 2-glycoprotein 1735:1767 leucine-rich alpha 2-glycoprotein 1735:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 1 100 theme sequence 330:337 arg1 analysis 339:346 amino acid sequence analysis 319:346 amino acid sequence analysis 319:346 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 1 101 theme cDNA 302:305 arg1 cloning 307:313 cDNA cloning 302:313 cDNA cloning 302:313 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 0 102 gly glycoprotein 44:55 arg1 glycoprotein 44:55 human platelet glycoprotein Ib 29:58 human platelet glycoprotein Ib 29:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 4 103 theme amino 878:882 arg1 acids 884:888 181 amino acids 874:888 181 amino acids 874:888 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 8 104 theme 2-glycoprotein 1754:1767 arg1 chain 1717:1721 the alpha chain 1707:1721 the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1707:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 5 105 theme polyadenylylation 994:1010 arg1 nucleotides 1031:1041 a polyadenylylation signal (AATAAA) 14 nucleotides 992:1041 a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides 992:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 0 106 theme alpha 4:8 arg1 proteins 83:90 both transmembrane proteins 64:90 both transmembrane proteins containing a leucine-rich amino acid sequence 64:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 0 106 theme alpha 4:8 arg1 chains 19:24 The alpha and beta chains 0:24 The alpha and beta chains of human platelet glycoprotein Ib 0:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 1 107 theme acid 325:328 arg1 sequence 330:337 amino acid sequence 319:337 amino acid sequence analysis 319:346 The primary structure of the beta chain of human glycoprotein Ib (GPIb), the platelet receptor for von Willebrand factor, has been established by a combination of cDNA cloning and amino acid sequence analysis. 3353370 9 108 theme acids 1873:1877 arg1 segment 1853:1859 a transmembrane segment 1837:1859 a transmembrane segment of 25 amino acids 1837:1877 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 9 108 theme acids 1873:1877 arg1 segment 1900:1906 an intracellular segment 1883:1906 an intracellular segment of 34 amino acids 1883:1924 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 0 109 theme beta 14:17 arg1 proteins 83:90 both transmembrane proteins 64:90 both transmembrane proteins containing a leucine-rich amino acid sequence 64:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 0 109 theme beta 14:17 arg1 chains 19:24 The alpha and beta chains 0:24 The alpha and beta chains of human platelet glycoprotein Ib 0:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 5 110 theme A 1066:1066 arg1 tail 1069:1072 the poly(A) tail 1057:1072 the poly(A) tail of 18 nucleotides 1057:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 8 111 theme leucine-rich 1676:1687 arg1 repeats 1696:1702 the leucine-rich tandem repeats 1672:1702 the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein 1672:1767 The leucine-rich sequence in the beta chain of GPIb is flanked on both sides by amino acid sequences that are similar to those flanking the leucine-rich tandem repeats of the alpha chain of GPIb and leucine-rich alpha 2-glycoprotein. 3353370 2 112 theme cDNA 364:367 arg1 library 380:386 A lambda phage cDNA expression library 349:386 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) 349:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 7 113 theme leucine-rich 1501:1512 arg1 2-glycoprotein 1520:1533 leucine-rich alpha 2-glycoprotein 1501:1533 leucine-rich alpha 2-glycoprotein 1501:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 4 114 theme open 768:771 arg1 frame 781:785 an open reading frame 765:785 an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon 765:902 The composite sequence spanned 968 nucleotides and included a 5' untranslated region of 22 nucleotides, an open reading frame of 618 nucleotides encoding a signal peptide of 28 amino acids and a mature protein of 181 amino acids, a stop codon, and a 3' noncoding region of 307 nucleotides. 3353370 0 115 contain containing 92:101 arg1 proteins 83:90 both transmembrane proteins 64:90 both transmembrane proteins containing a leucine-rich amino acid sequence 64:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 0 115 contain containing 92:101 arg2 sequence 129:136 a leucine-rich amino acid sequence 103:136 a leucine-rich amino acid sequence 103:136 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 0 115 contain containing 92:101 arg1 chains 19:24 The alpha and beta chains 0:24 The alpha and beta chains of human platelet glycoprotein Ib 0:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 2 116 theme lambda 351:356 arg1 library 380:386 A lambda phage cDNA expression library 349:386 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) 349:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 0 117 theme platelet 35:42 arg1 Ib 57:58 human platelet glycoprotein Ib 29:58 human platelet glycoprotein Ib 29:58 The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. 3353370 5 118 theme signal 1012:1017 arg1 nucleotides 1031:1041 a polyadenylylation signal (AATAAA) 14 nucleotides 992:1041 a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides 992:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 2 119 theme erythroleukemia 408:422 arg1 cells 424:428 human erythroleukemia cells 402:428 human erythroleukemia cells (HEL cells) 402:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 2 119 theme erythroleukemia 408:422 arg1 cells 435:439 HEL cells 431:439 HEL cells 431:439 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 3 120 theme three 638:642 arg1 sequences 625:633 the nucleotide sequences 610:633 the nucleotide sequences of three 610:642 Eighteen positive clones were isolated and plaque-purified and the nucleotide sequences of three were determined. 3353370 5 121 theme poly 1061:1064 arg1 tail 1069:1072 the poly(A) tail 1057:1072 the poly(A) tail of 18 nucleotides 1057:1090 The 3' noncoding sequence also contained a polyadenylylation signal (AATAAA) 14 nucleotides upstream from the poly(A) tail of 18 nucleotides. 3353370 6 122 theme acid 1196:1199 arg1 sequences 1201:1209 amino acid sequences 1190:1209 amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA 1190:1283 Edman degradation of the intact beta chain and of peptides produced by chemical cleavage yielded amino acid sequences spanning 76 residues that were identical to those predicted from the cDNA. 3353370 9 123 theme acids 1920:1924 arg1 segment 1853:1859 a transmembrane segment 1837:1859 a transmembrane segment of 25 amino acids 1837:1877 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 9 123 theme acids 1920:1924 arg1 segment 1900:1906 an intracellular segment 1883:1906 an intracellular segment of 34 amino acids 1883:1924 The amino-terminal region of the beta chain of GPIb is followed by a transmembrane segment of 25 amino acids and an intracellular segment of 34 amino acids at the carboxyl terminus of the protein. 3353370 7 124 theme acids 1375:1379 arg1 sequence 1354:1361 a leucine-rich sequence 1339:1361 a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein 1339:1533 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 7 124 theme acids 1375:1379 arg1 similar 1389:1395 similar 1389:1395 similar 1389:1395 The amino-terminal region of the beta chain contains a leucine-rich sequence of 24 amino acids that is similar to a sequence that occurs as seven tandem repeats in the alpha chain of GPIb and nine tandem repeats in leucine-rich alpha 2-glycoprotein. 3353370 2 125 theme human 402:406 arg1 cells 424:428 human erythroleukemia cells 402:428 human erythroleukemia cells (HEL cells) 402:440 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 3353370 2 125 theme human 402:406 arg1 cells 435:439 HEL cells 431:439 HEL cells 431:439 A lambda phage cDNA expression library prepared from human erythroleukemia cells (HEL cells) was screened with a radiolabeled affinity-purified rabbit polyclonal antibody to the beta chain of GPIb. 29217581 2 0 theme TRP 317:319 arg1 channels 321:328 many other TRP channels 306:328 many other TRP channels 306:328 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 6 1 theme voltage-dependent 894:910 arg1 opening 912:918 voltage-dependent opening 894:918 voltage-dependent opening 894:918 Calcium binding induces conformational changes that likely prime the channel for voltage-dependent opening. 29217581 2 2 theme other 311:315 arg1 channels 321:328 many other TRP channels 306:328 many other TRP channels 306:328 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 0 3 from Structure 0:8 arg1 nanodisc 52:59 a lipid nanodisc 44:59 a lipid nanodisc 44:59 Structure of the human TRPM4 ion channel in a lipid nanodisc. 29217581 4 4 theme intracellular 716:728 arg1 side 730:733 the intracellular side 712:733 the intracellular side of the S1-S4 domain 712:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 3 5 theme single-particle 502:516 arg1 microscopy 532:541 single-particle cryo-electron microscopy 502:541 single-particle cryo-electron microscopy 502:541 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 3 6 from resolution 473:482 arg1 nanodiscs 448:456 lipid nanodiscs 442:456 lipid nanodiscs at ~3-angstrom resolution 442:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 2 7 theme intracellular 242:254 arg1 calcium 256:262 increased intracellular calcium 232:262 increased intracellular calcium 232:262 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 4 8 theme channels 656:663 arg1 subfamily 639:647 this major subfamily 628:647 this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain 628:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 3 9 theme lipid 442:446 arg1 nanodiscs 448:456 lipid nanodiscs 442:456 lipid nanodiscs at ~3-angstrom resolution 442:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 2 10 theme increased 232:240 arg1 calcium 256:262 increased intracellular calcium 232:262 increased intracellular calcium 232:262 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 1 11 theme expressed 130:138 arg1 melastatin 97:106 Transient receptor potential (TRP) melastatin 4 62:108 Transient receptor potential (TRP) melastatin 4 (TRPM4) 62:116 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 11 theme expressed 130:138 arg1 channel 147:153 a widely expressed cation channel 121:153 a widely expressed cation channel associated with a variety of cardiovascular disorders 121:207 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 2 12 theme monovalent 347:356 arg1 cations 358:364 monovalent cations 347:364 monovalent cations 347:364 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 3 13 from embedded 430:437 arg1 nanodiscs 448:456 lipid nanodiscs 442:456 lipid nanodiscs at ~3-angstrom resolution 442:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 4 14 theme S1-S4 742:746 arg1 domain 748:753 the S1-S4 domain 738:753 the S1-S4 domain 738:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 1 15 theme Transient 62:70 arg1 TRP 92:94 TRP 92:94 TRP 92:94 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 15 theme Transient 62:70 arg1 potential 81:89 Transient receptor potential 62:89 Transient receptor potential (TRP) melastatin 4 (TRPM4) 62:116 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 2 16 dep activated 219:227 arg1 permeable 334:342 permeable 334:342 permeable 334:342 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 2 17 theme voltage-dependent 269:285 arg1 manner 287:292 a voltage-dependent manner 267:292 a voltage-dependent manner 267:292 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 1 18 theme cation 140:145 arg1 melastatin 97:106 Transient receptor potential (TRP) melastatin 4 62:108 Transient receptor potential (TRP) melastatin 4 (TRPM4) 62:116 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 18 theme cation 140:145 arg1 channel 147:153 a widely expressed cation channel 121:153 a widely expressed cation channel associated with a variety of cardiovascular disorders 121:207 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 0 19 theme human 17:21 arg1 channel 33:39 the human TRPM4 ion channel 13:39 the human TRPM4 ion channel 13:39 Structure of the human TRPM4 ion channel in a lipid nanodisc. 29217581 3 20 from nanodiscs 448:456 arg1 embedded 430:437 embedded 430:437 embedded 430:437 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 1 21 theme potential 81:89 arg1 melastatin 97:106 Transient receptor potential (TRP) melastatin 4 62:108 Transient receptor potential (TRP) melastatin 4 (TRPM4) 62:116 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 21 theme potential 81:89 arg1 TRPM4 111:115 TRPM4 111:115 TRPM4 111:115 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 21 theme potential 81:89 arg1 channel 147:153 a widely expressed cation channel 121:153 a widely expressed cation channel associated with a variety of cardiovascular disorders 121:207 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 3 22 theme human 418:422 arg1 TRPM4 424:428 full-length human TRPM4 406:428 full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution 406:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 1 23 theme receptor 72:79 arg1 TRP 92:94 TRP 92:94 TRP 92:94 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 23 theme receptor 72:79 arg1 potential 81:89 Transient receptor potential 62:89 Transient receptor potential (TRP) melastatin 4 (TRPM4) 62:116 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 4 24 theme domain 748:753 arg1 side 730:733 the intracellular side 712:733 the intracellular side of the S1-S4 domain 712:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 3 25 theme ~3-angstrom 461:471 arg1 resolution 473:482 ~3-angstrom resolution 461:482 ~3-angstrom resolution 461:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 3 26 theme TRPM4 424:428 arg1 structures 392:401 two structures 388:401 two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution 388:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 0 27 theme ion 29:31 arg1 channel 33:39 the human TRPM4 ion channel 13:39 the human TRPM4 ion channel 13:39 Structure of the human TRPM4 ion channel in a lipid nanodisc. 29217581 4 28 theme major 633:637 arg1 subfamily 639:647 this major subfamily 628:647 this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain 628:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 0 29 theme TRPM4 23:27 arg1 channel 33:39 the human TRPM4 ion channel 13:39 the human TRPM4 ion channel 13:39 Structure of the human TRPM4 ion channel in a lipid nanodisc. 29217581 6 30 theme conformational 837:850 arg1 changes 852:858 conformational changes 837:858 conformational changes that likely prime the channel for voltage-dependent opening 837:918 Calcium binding induces conformational changes that likely prime the channel for voltage-dependent opening. 29217581 5 31 theme distinct 789:796 arg1 states 805:810 two distinct closed states 785:810 two distinct closed states 785:810 The structures correspond to two distinct closed states. 29217581 3 32 theme full-length 406:416 arg1 TRPM4 424:428 full-length human TRPM4 406:428 full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution 406:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 2 33 theme many 306:309 arg1 channels 321:328 many other TRP channels 306:328 many other TRP channels 306:328 TRPM4 is activated by increased intracellular calcium in a voltage-dependent manner but, unlike many other TRP channels, is permeable to monovalent cations only. 29217581 0 34 theme channel 33:39 arg1 Structure 0:8 Structure 0:8 Structure of the human TRPM4 ion channel in a lipid nanodisc. 0:60 Structure of the human TRPM4 ion channel in a lipid nanodisc. 29217581 4 35 theme well-defined 671:682 arg1 site 700:703 a well-defined calcium-binding site 669:703 a well-defined calcium-binding site within the intracellular side of the S1-S4 domain 669:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 6 36 theme Calcium 813:819 arg1 binding 821:827 Calcium binding 813:827 Calcium binding 813:827 Calcium binding induces conformational changes that likely prime the channel for voltage-dependent opening. 29217581 3 37 theme embedded 430:437 arg1 TRPM4 424:428 full-length human TRPM4 406:428 full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution 406:482 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 3 38 theme cryo-electron 518:530 arg1 microscopy 532:541 single-particle cryo-electron microscopy 502:541 single-particle cryo-electron microscopy 502:541 Here we present two structures of full-length human TRPM4 embedded in lipid nanodiscs at ~3-angstrom resolution, as determined by single-particle cryo-electron microscopy. 29217581 4 39 theme calcium-binding 684:698 arg1 site 700:703 a well-defined calcium-binding site 669:703 a well-defined calcium-binding site within the intracellular side of the S1-S4 domain 669:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 4 40 theme site 700:703 arg1 subfamily 639:647 this major subfamily 628:647 this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain 628:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 0 41 theme lipid 46:50 arg1 nanodisc 52:59 a lipid nanodisc 44:59 a lipid nanodisc 44:59 Structure of the human TRPM4 ion channel in a lipid nanodisc. 29217581 4 42 theme general 603:609 arg1 architecture 611:622 a general architecture 601:622 a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain 601:753 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 4 43 theme TRP 652:654 arg1 channels 656:663 TRP channels 652:663 TRP channels 652:663 These structures, with and without calcium bound, reveal a general architecture for this major subfamily of TRP channels and a well-defined calcium-binding site within the intracellular side of the S1-S4 domain. 29217581 1 44 theme cardiovascular 184:197 arg1 disorders 199:207 cardiovascular disorders 184:207 cardiovascular disorders 184:207 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 5 45 theme closed 798:803 arg1 states 805:810 two distinct closed states 785:810 two distinct closed states 785:810 The structures correspond to two distinct closed states. 29217581 1 46 theme disorders 199:207 arg1 variety 173:179 a variety 171:179 a variety of cardiovascular disorders 171:207 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 29217581 1 46 theme disorders 199:207 arg1 disorders 199:207 cardiovascular disorders 184:207 cardiovascular disorders 184:207 Transient receptor potential (TRP) melastatin 4 (TRPM4) is a widely expressed cation channel associated with a variety of cardiovascular disorders. 32589945 0 0 theme Receptor 86:93 arg1 Specificity 95:105 Receptor Specificity 86:105 Receptor Specificity 86:105 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 9 1 theme related 1221:1227 arg1 toxins 1229:1234 highly related toxins 1214:1234 highly related toxins 1214:1234 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 7 2 theme Å 771:771 arg1 structure 781:789 A 3.3 Å cryo-EM structure 765:789 A 3.3 Å cryo-EM structure 765:789 A 3.3 Å cryo-EM structure shows that TcsL binds SEMA6A with the same region that in TcdB binds structurally unrelated Frizzled. 32589945 3 3 theme similar 379:385 arg1 % 377:377 87% 375:377 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium 375:474 TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. 32589945 8 4 theme binding 984:990 arg1 specificity 992:1002 binding specificity 984:1002 binding specificity of TcsL 984:1010 Remarkably, 15 mutations in this evolutionarily divergent surface are sufficient to switch binding specificity of TcsL to that of TcdB. 32589945 2 5 theme shock 310:314 arg1 syndrome 316:323 an almost invariably lethal toxic shock syndrome 276:323 an almost invariably lethal toxic shock syndrome associated with gynecological infections 276:364 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 0 6 theme Specificity 95:105 arg1 Principles 72:81 Principles 72:81 Principles of Receptor Specificity in Clostridial Toxins 72:127 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 2 7 theme toxic 304:308 arg1 syndrome 316:323 an almost invariably lethal toxic shock syndrome 276:323 an almost invariably lethal toxic shock syndrome associated with gynecological infections 276:364 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 8 8 theme divergent 941:949 arg1 surface 951:957 this evolutionarily divergent surface 921:957 this evolutionarily divergent surface 921:957 Remarkably, 15 mutations in this evolutionarily divergent surface are sufficient to switch binding specificity of TcsL to that of TcdB. 32589945 4 9 theme P. sordellii 486:497 arg1 infections 499:508 P. sordellii infections 486:508 P. sordellii infections 486:508 However, P. sordellii infections target vascular endothelium, suggesting that TcsL exploits another receptor. 32589945 2 10 theme lethal 297:302 arg1 syndrome 316:323 an almost invariably lethal toxic shock syndrome 276:323 an almost invariably lethal toxic shock syndrome associated with gynecological infections 276:364 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 3 11 theme Frizzled 437:444 arg1 receptors 446:454 Frizzled receptors 437:454 Frizzled receptors in colon epithelium 437:474 TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. 32589945 0 12 theme Clostridial 110:120 arg1 Toxins 122:127 Clostridial Toxins 110:127 Clostridial Toxins 110:127 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 4 13 theme vascular 517:524 arg1 endothelium 526:536 vascular endothelium 517:536 vascular endothelium 517:536 However, P. sordellii infections target vascular endothelium, suggesting that TcsL exploits another receptor. 32589945 5 14 theme CRISPR/Cas9 599:609 arg1 screening 611:619 CRISPR/Cas9 screening 599:619 CRISPR/Cas9 screening 599:619 Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. 32589945 7 15 theme unrelated 873:881 arg1 Frizzled 883:890 structurally unrelated Frizzled 860:890 structurally unrelated Frizzled 860:890 A 3.3 Å cryo-EM structure shows that TcsL binds SEMA6A with the same region that in TcdB binds structurally unrelated Frizzled. 32589945 1 16 theme severe 195:200 arg1 damage 214:219 severe host tissue damage 195:219 severe host tissue damage 195:219 Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. 32589945 7 17 theme same 829:832 arg1 region 834:839 the same region 825:839 the same region that in TcdB binds structurally unrelated Frizzled 825:890 A 3.3 Å cryo-EM structure shows that TcsL binds SEMA6A with the same region that in TcdB binds structurally unrelated Frizzled. 32589945 5 18 theme TcsL 668:671 arg1 semaphorins 635:645 semaphorins SEMA6A and SEMA6B 635:663 semaphorins SEMA6A and SEMA6B 635:663 Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. 32589945 5 18 theme TcsL 668:671 arg1 receptors 673:681 TcsL receptors 668:681 TcsL receptors 668:681 Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. 32589945 2 19 theme gynecological 341:353 arg1 infections 355:364 gynecological infections 341:364 gynecological infections 341:364 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 1 20 theme host 202:205 arg1 damage 214:219 severe host tissue damage 195:219 severe host tissue damage 195:219 Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. 32589945 1 21 theme tissue 207:212 arg1 damage 214:219 severe host tissue damage 195:219 severe host tissue damage 195:219 Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. 32589945 3 22 theme C. difficile 390:401 arg1 TcdB 403:406 C. difficile TcdB 390:406 C. difficile TcdB 390:406 TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. 32589945 0 23 theme Semaphorin 15:24 arg1 Proteins 26:33 Semaphorin Proteins 15:33 Semaphorin Proteins 15:33 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 6 24 theme recombinant 704:714 arg1 SEMA6A 716:721 recombinant SEMA6A 704:721 recombinant SEMA6A 704:721 We demonstrate that recombinant SEMA6A can protect mice from TcsL-induced edema. 32589945 0 25 theme P. sordellii 38:49 arg1 Toxin 58:62 P. sordellii Lethal Toxin 38:62 P. sordellii Lethal Toxin 38:62 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 6 26 theme TcsL-induced 745:756 arg1 edema 758:762 TcsL-induced edema 745:762 TcsL-induced edema 745:762 We demonstrate that recombinant SEMA6A can protect mice from TcsL-induced edema. 32589945 9 27 from difference 1150:1159 arg1 targeting 1171:1179 tissue targeting 1164:1179 tissue targeting 1164:1179 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 9 27 from difference 1150:1159 arg1 pathogenesis 1193:1204 disease pathogenesis 1185:1204 disease pathogenesis 1185:1204 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 5 28 dep semaphorins 635:645 arg1 semaphorins 635:645 semaphorins SEMA6A and SEMA6B 635:663 semaphorins SEMA6A and SEMA6B 635:663 Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. 32589945 5 28 dep semaphorins 635:645 arg1 SEMA6B 658:663 SEMA6B 658:663 SEMA6B 658:663 Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. 32589945 5 28 dep semaphorins 635:645 arg1 SEMA6A 647:652 SEMA6A 647:652 SEMA6A 647:652 Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. 32589945 0 29 from Principles 72:81 arg1 Toxins 122:127 Clostridial Toxins 110:127 Clostridial Toxins 110:127 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 0 30 theme Proteins 26:33 arg1 Recognition 0:10 Recognition 0:10 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin 0:62 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 9 31 theme tissue 1164:1169 arg1 targeting 1171:1179 tissue targeting 1164:1179 tissue targeting 1164:1179 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 0 32 theme Lethal 51:56 arg1 Toxin 58:62 P. sordellii Lethal Toxin 38:62 P. sordellii Lethal Toxin 38:62 Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins. 32589945 2 33 theme lethal 249:254 arg1 TcsL 263:266 TcsL 263:266 TcsL 263:266 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 2 33 theme lethal 249:254 arg1 toxin 256:260 Paeniclostridium sordellii lethal toxin 222:260 Paeniclostridium sordellii lethal toxin (TcsL) 222:267 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 1 34 theme Pathogenic 130:139 arg1 species 153:159 Pathogenic clostridial species 130:159 Pathogenic clostridial species 130:159 Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. 32589945 2 35 theme Paeniclostridium 222:237 arg1 TcsL 263:266 TcsL 263:266 TcsL 263:266 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 2 35 theme Paeniclostridium 222:237 arg1 toxin 256:260 Paeniclostridium sordellii lethal toxin 222:260 Paeniclostridium sordellii lethal toxin (TcsL) 222:267 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 32589945 8 36 from mutations 908:916 arg1 surface 951:957 this evolutionarily divergent surface 921:957 this evolutionarily divergent surface 921:957 Remarkably, 15 mutations in this evolutionarily divergent surface are sufficient to switch binding specificity of TcsL to that of TcdB. 32589945 1 37 theme clostridial 141:151 arg1 species 153:159 Pathogenic clostridial species 130:159 Pathogenic clostridial species 130:159 Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. 32589945 9 38 theme disease 1185:1191 arg1 pathogenesis 1193:1204 disease pathogenesis 1185:1204 disease pathogenesis 1185:1204 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 9 39 theme molecular 1126:1134 arg1 basis 1136:1140 the molecular basis 1122:1140 the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins 1122:1234 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 3 40 from receptors 446:454 arg1 epithelium 465:474 colon epithelium 459:474 colon epithelium 459:474 TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. 32589945 3 41 theme host 422:425 arg1 cells 427:431 host cells 422:431 host cells 422:431 TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. 32589945 3 42 theme colon 459:463 arg1 epithelium 465:474 colon epithelium 459:474 colon epithelium 459:474 TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. 32589945 7 43 theme cryo-EM 773:779 arg1 structure 781:789 A 3.3 Å cryo-EM structure 765:789 A 3.3 Å cryo-EM structure 765:789 A 3.3 Å cryo-EM structure shows that TcsL binds SEMA6A with the same region that in TcdB binds structurally unrelated Frizzled. 32589945 9 44 theme relevant 1083:1090 arg1 semaphorins 1052:1062 semaphorins 1052:1062 semaphorins 1052:1062 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 9 44 theme relevant 1083:1090 arg1 receptors 1092:1100 physiologically relevant receptors 1067:1100 physiologically relevant receptors for TcsL 1067:1109 Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins. 32589945 8 45 theme TcsL 1007:1010 arg1 specificity 992:1002 binding specificity 984:1002 binding specificity of TcsL 984:1010 Remarkably, 15 mutations in this evolutionarily divergent surface are sufficient to switch binding specificity of TcsL to that of TcdB. 32589945 1 46 theme potent 169:174 arg1 toxins 176:181 potent toxins 169:181 potent toxins that induce severe host tissue damage 169:219 Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. 32589945 2 47 dep Paeniclostridium 222:237 arg1 sordellii 239:247 sordellii 239:247 sordellii 239:247 Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. 26371182 8 0 theme ATX 1110:1112 arg1 inhibitors 1114:1123 novel ATX inhibitors 1104:1123 novel ATX inhibitors 1104:1123 Our studies may provide a basis for the rational design of novel ATX inhibitors. 26371182 7 1 theme inhibition 968:977 arg1 mechanism 955:963 the mechanism 951:963 the mechanism of inhibition of each compound 951:994 We demonstrate that the mechanism of inhibition of each compound reflects its unique interactions with human ATX. 26371182 8 2 theme rational 1085:1092 arg1 design 1094:1099 the rational design 1081:1099 the rational design of novel ATX inhibitors 1081:1123 Our studies may provide a basis for the rational design of novel ATX inhibitors. 26371182 8 3 theme inhibitors 1114:1123 arg1 design 1094:1099 the rational design 1081:1099 the rational design of novel ATX inhibitors 1081:1123 Our studies may provide a basis for the rational design of novel ATX inhibitors. 26371182 1 4 theme secreted 129:136 arg1 enzyme 138:143 a secreted enzyme 127:143 a secreted enzyme that hydrolyzes lysophosphatidylcholine to lysophosphatidic acid (LPA) 127:214 Autotaxin (ATX) is a secreted enzyme that hydrolyzes lysophosphatidylcholine to lysophosphatidic acid (LPA). 26371182 1 4 theme secreted 129:136 arg1 Autotaxin 108:116 Autotaxin 108:116 Autotaxin (ATX) 108:122 Autotaxin (ATX) is a secreted enzyme that hydrolyzes lysophosphatidylcholine to lysophosphatidic acid (LPA). 26371182 6 5 dep unpublished 880:890 arg1 distinct 906:913 distinct 906:913 distinct 906:913 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 6 6 theme unpublished 880:890 arg1 inhibitors 919:928 four previously unpublished, structurally distinct ATX inhibitors 864:928 four previously unpublished, structurally distinct ATX inhibitors 864:928 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 0 7 with Compounds 66:74 arg1 Modes 90:94 Distinct Modes 81:94 Distinct Modes of Binding 81:105 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 2 8 theme biological 272:281 arg1 migration 324:332 migration 324:332 migration 324:332 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 8 theme biological 272:281 arg1 survival/apoptosis 339:356 survival/apoptosis 339:356 survival/apoptosis 339:356 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 8 theme biological 272:281 arg1 proliferation 309:321 cell proliferation 304:321 cell proliferation 304:321 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 8 theme biological 272:281 arg1 processes 283:291 diverse biological processes 264:291 diverse biological processes 264:291 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 0 9 theme Distinct 81:88 arg1 Modes 90:94 Distinct Modes 81:94 Distinct Modes of Binding 81:105 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 2 10 theme diverse 264:270 arg1 migration 324:332 migration 324:332 migration 324:332 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 10 theme diverse 264:270 arg1 survival/apoptosis 339:356 survival/apoptosis 339:356 survival/apoptosis 339:356 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 10 theme diverse 264:270 arg1 proliferation 309:321 cell proliferation 304:321 cell proliferation 304:321 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 10 theme diverse 264:270 arg1 processes 283:291 diverse biological processes 264:291 diverse biological processes 264:291 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 0 11 theme Binding 99:105 arg1 Modes 90:94 Distinct Modes 81:94 Distinct Modes of Binding 81:105 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 4 12 theme attractive 610:619 arg1 strategy 621:628 an attractive strategy 607:628 an attractive strategy for the development of therapeutics to treat a variety of diseases 607:695 Therefore, ATX inhibitors represent an attractive strategy for the development of therapeutics to treat a variety of diseases. 26371182 7 13 with interactions 1016:1027 arg1 ATX 1040:1042 human ATX 1034:1042 human ATX 1034:1042 We demonstrate that the mechanism of inhibition of each compound reflects its unique interactions with human ATX. 26371182 3 14 theme ATX-LPA 430:436 arg1 pathway 438:444 The ATX-LPA pathway 426:444 The ATX-LPA pathway 426:444 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 5 15 dep Mouse 698:702 arg1 ATX 712:714 ATX 712:714 ATX 712:714 Mouse and rat ATX have been crystallized previously with LPA or small-molecule inhibitors bound. 26371182 2 16 theme cell 304:307 arg1 proliferation 309:321 cell proliferation 304:321 cell proliferation 304:321 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 0 17 theme Structural 0:9 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 0:106 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 4 18 theme therapeutics 653:664 arg1 development 638:648 the development 634:648 the development of therapeutics 634:664 Therefore, ATX inhibitors represent an attractive strategy for the development of therapeutics to treat a variety of diseases. 26371182 3 19 theme cholestatic 539:549 arg1 pruritus 551:558 cholestatic pruritus 539:558 cholestatic pruritus 539:558 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 2 20 theme protein-coupled 399:413 arg1 receptors 415:423 G protein-coupled receptors 397:423 G protein-coupled receptors 397:423 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 21 theme receptors 415:423 arg1 family 387:392 a family 385:392 a family of G protein-coupled receptors 385:423 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 1 22 theme lysophosphatidic 188:203 arg1 LPA 211:213 LPA 211:213 LPA 211:213 Autotaxin (ATX) is a secreted enzyme that hydrolyzes lysophosphatidylcholine to lysophosphatidic acid (LPA). 26371182 1 22 theme lysophosphatidic 188:203 arg1 acid 205:208 lysophosphatidic acid 188:208 lysophosphatidic acid (LPA) 188:214 Autotaxin (ATX) is a secreted enzyme that hydrolyzes lysophosphatidylcholine to lysophosphatidic acid (LPA). 26371182 0 23 theme Human 35:39 arg1 Autotaxin 41:49 Human Autotaxin 35:49 Human Autotaxin 35:49 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 6 24 theme ATX 844:846 arg1 structures 824:833 the crystal structures 812:833 the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors 812:928 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 5 25 theme small-molecule 762:775 arg1 inhibitors 777:786 small-molecule inhibitors 762:786 small-molecule inhibitors bound 762:792 Mouse and rat ATX have been crystallized previously with LPA or small-molecule inhibitors bound. 26371182 6 26 theme human 838:842 arg1 ATX 844:846 human ATX 838:846 human ATX 838:846 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 2 27 theme family 387:392 arg1 activation 371:380 the activation 367:380 the activation of a family of G protein-coupled receptors 367:423 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 6 28 from structures 824:833 arg1 complex 851:857 complex 851:857 complex with four previously unpublished, structurally distinct ATX inhibitors 851:928 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 7 29 theme human 1034:1038 arg1 ATX 1040:1042 human ATX 1034:1042 human ATX 1034:1042 We demonstrate that the mechanism of inhibition of each compound reflects its unique interactions with human ATX. 26371182 7 30 theme unique 1009:1014 arg1 interactions 1016:1027 its unique interactions 1005:1027 its unique interactions with human ATX 1005:1042 We demonstrate that the mechanism of inhibition of each compound reflects its unique interactions with human ATX. 26371182 3 31 theme pathologic 474:483 arg1 cancer 507:512 cancer 507:512 cancer 507:512 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 31 theme pathologic 474:483 arg1 pruritus 551:558 cholestatic pruritus 539:558 cholestatic pruritus 539:558 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 31 theme pathologic 474:483 arg1 fibrosis 515:522 fibrosis 515:522 fibrosis 515:522 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 31 theme pathologic 474:483 arg1 conditions 485:494 many pathologic conditions 469:494 many pathologic conditions 469:494 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 31 theme pathologic 474:483 arg1 inflammation 525:536 inflammation 525:536 inflammation 525:536 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 31 theme pathologic 474:483 arg1 pain 565:568 pain 565:568 pain 565:568 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 0 32 theme Autotaxin 41:49 arg1 Inhibition 21:30 Inhibition 21:30 Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding 21:105 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 7 33 theme compound 987:994 arg1 inhibition 968:977 inhibition 968:977 inhibition of each compound 968:994 We demonstrate that the mechanism of inhibition of each compound reflects its unique interactions with human ATX. 26371182 4 34 theme diseases 688:695 arg1 variety 677:683 a variety 675:683 a variety of diseases 675:695 Therefore, ATX inhibitors represent an attractive strategy for the development of therapeutics to treat a variety of diseases. 26371182 4 34 theme diseases 688:695 arg1 diseases 688:695 diseases 688:695 diseases 688:695 Therefore, ATX inhibitors represent an attractive strategy for the development of therapeutics to treat a variety of diseases. 26371182 6 35 theme ATX 915:917 arg1 inhibitors 919:928 four previously unpublished, structurally distinct ATX inhibitors 864:928 four previously unpublished, structurally distinct ATX inhibitors 864:928 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 6 36 theme crystal 816:822 arg1 structures 824:833 the crystal structures 812:833 the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors 812:928 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26371182 2 37 theme bioactive 226:234 arg1 LPA 217:219 LPA 217:219 LPA 217:219 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 2 37 theme bioactive 226:234 arg1 phospholipid 236:247 a bioactive phospholipid 224:247 a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors 224:423 LPA is a bioactive phospholipid that regulates diverse biological processes, including cell proliferation, migration, and survival/apoptosis, through the activation of a family of G protein-coupled receptors. 26371182 3 38 theme many 469:472 arg1 cancer 507:512 cancer 507:512 cancer 507:512 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 38 theme many 469:472 arg1 pruritus 551:558 cholestatic pruritus 539:558 cholestatic pruritus 539:558 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 38 theme many 469:472 arg1 fibrosis 515:522 fibrosis 515:522 fibrosis 515:522 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 38 theme many 469:472 arg1 conditions 485:494 many pathologic conditions 469:494 many pathologic conditions 469:494 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 38 theme many 469:472 arg1 inflammation 525:536 inflammation 525:536 inflammation 525:536 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 3 38 theme many 469:472 arg1 pain 565:568 pain 565:568 pain 565:568 The ATX-LPA pathway has been implicated in many pathologic conditions, including cancer, fibrosis, inflammation, cholestatic pruritus, and pain. 26371182 4 39 theme ATX 582:584 arg1 inhibitors 586:595 ATX inhibitors 582:595 ATX inhibitors 582:595 Therefore, ATX inhibitors represent an attractive strategy for the development of therapeutics to treat a variety of diseases. 26371182 0 40 theme Potent 59:64 arg1 Compounds 66:74 Four Potent Compounds 54:74 Four Potent Compounds with Distinct Modes of Binding 54:105 Structural Basis for Inhibition of Human Autotaxin by Four Potent Compounds with Distinct Modes of Binding. 26371182 8 41 theme novel 1104:1108 arg1 inhibitors 1114:1123 novel ATX inhibitors 1104:1123 novel ATX inhibitors 1104:1123 Our studies may provide a basis for the rational design of novel ATX inhibitors. 26371182 6 42 with complex 851:857 arg1 inhibitors 919:928 four previously unpublished, structurally distinct ATX inhibitors 864:928 four previously unpublished, structurally distinct ATX inhibitors 864:928 Here, we present the crystal structures of human ATX in complex with four previously unpublished, structurally distinct ATX inhibitors. 26811476 7 0 gly glycoprotein 1322:1333 arg1 glycoproteins 1285:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins 1235:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1235:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 0 gly glycoprotein 1322:1333 arg1 glycoprotein 1322:1333 glycoprotein 2 1322:1335 glycoprotein 2 (GP2) 1322:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 0 gly glycoprotein 1322:1333 arg1 GP2 1338:1340 GP2 1338:1340 GP2 1338:1340 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 6 1 theme domain 1047:1052 arg1 homodimerization 1054:1069 ZP-N domain homodimerization 1042:1069 ZP-N domain homodimerization 1042:1069 The structure of UMOD reveals that an extensive hydrophobic interface mediates ZP-N domain homodimerization. 26811476 2 2 theme extracellular 327:339 arg1 filaments 341:349 extracellular filaments 327:349 extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones 327:451 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 1 3 theme therapeutic 226:236 arg1 target 238:243 a promising therapeutic target 214:243 a promising therapeutic target for hypertension 214:260 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 3 4 theme UMOD 494:497 arg1 filaments 499:507 UMOD filaments 494:507 UMOD filaments 494:507 Moreover, salt-dependent aggregation of UMOD filaments in the urine generates a soluble molecular net that captures uropathogenic bacteria and facilitates their clearance. 26811476 9 5 theme pathogenic 1538:1547 arg1 mutations 1549:1557 numerous pathogenic mutations 1529:1557 numerous pathogenic mutations 1529:1557 Moreover, the architecture of UMOD rationalizes numerous pathogenic mutations in both UMOD and TECTA genes. 26811476 3 6 theme filaments 499:507 arg1 aggregation 479:489 salt-dependent aggregation 464:489 salt-dependent aggregation of UMOD filaments in the urine 464:520 Moreover, salt-dependent aggregation of UMOD filaments in the urine generates a soluble molecular net that captures uropathogenic bacteria and facilitates their clearance. 26811476 2 7 theme bipartite 271:279 arg1 pellucida 286:294 its bipartite zona pellucida module (ZP-N/ZP-C) 267:313 its bipartite zona pellucida module (ZP-N/ZP-C) 267:313 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 4 8 dep available 710:718 arg1 self-assembles 739:752 self-assembles 739:752 self-assembles into filaments 739:767 Despite the functional importance of its homopolymers, no structural information is available on UMOD and how it self-assembles into filaments. 26811476 6 9 theme UMOD 980:983 arg1 structure 967:975 The structure 963:975 The structure of UMOD 963:983 The structure of UMOD reveals that an extensive hydrophobic interface mediates ZP-N domain homodimerization. 26811476 7 10 dep glycoproteins 1285:1297 arg1 GP2 1338:1340 GP2 1338:1340 GP2 1338:1340 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 10 dep glycoproteins 1285:1297 arg1 TECTA 1311:1315 TECTA 1311:1315 TECTA 1311:1315 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 10 dep glycoproteins 1285:1297 arg1 glycoprotein 1322:1333 glycoprotein 2 1322:1335 glycoprotein 2 (GP2) 1322:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 10 dep glycoproteins 1285:1297 arg1 α-tectorin 1299:1308 α-tectorin 1299:1308 α-tectorin (TECTA) 1299:1316 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 10 dep glycoproteins 1285:1297 arg1 glycoproteins 1285:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins 1235:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1235:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 5 11 theme receptor 883:890 arg1 UMOD 844:847 human UMOD 838:847 human UMOD 838:847 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 11 theme receptor 883:890 arg1 protein 892:898 an essential sperm receptor protein 864:898 an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 864:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 11 theme receptor 883:890 arg1 related 921:927 related 921:927 related 921:927 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 12 theme UMOD 844:847 arg1 regions 827:833 polymerization regions 812:833 polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 812:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 3 13 theme salt-dependent 464:477 arg1 aggregation 479:489 salt-dependent aggregation 464:489 salt-dependent aggregation of UMOD filaments in the urine 464:520 Moreover, salt-dependent aggregation of UMOD filaments in the urine generates a soluble molecular net that captures uropathogenic bacteria and facilitates their clearance. 26811476 7 14 theme filament 1105:1112 arg1 formation 1114:1122 filament formation 1105:1122 filament formation 1105:1122 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 5 15 theme human 838:842 arg1 UMOD 844:847 human UMOD 838:847 human UMOD 838:847 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 15 theme human 838:842 arg1 protein 892:898 an essential sperm receptor protein 864:898 an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 864:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 15 theme human 838:842 arg1 related 921:927 related 921:927 related 921:927 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 1 16 theme key 171:173 arg1 role 175:178 a key role 169:178 a key role 169:178 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 5 17 theme mouse 853:857 arg1 ZP2 859:861 mouse ZP2 853:861 mouse ZP2 853:861 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 7 18 theme ordered 1146:1152 arg1 linker 1164:1169 an ordered ZP-N/ZP-C linker 1143:1169 an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1143:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 19 located observed 1183:1190 arg2 linker 1164:1169 an ordered ZP-N/ZP-C linker 1143:1169 an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1143:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 19 located observed 1183:1190 arg1 ZP2 1195:1197 ZP2 1195:1197 ZP2 1195:1197 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 9 20 theme TECTA 1576:1580 arg1 genes 1582:1586 TECTA genes 1576:1586 TECTA genes 1576:1586 Moreover, the architecture of UMOD rationalizes numerous pathogenic mutations in both UMOD and TECTA genes. 26811476 2 21 theme electrolyte 372:382 arg1 balance 384:390 kidney electrolyte balance 365:390 kidney electrolyte balance 365:390 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 0 22 theme structured 2:11 arg1 linker 25:30 A structured interdomain linker 0:30 A structured interdomain linker 0:30 A structured interdomain linker directs self-polymerization of human uromodulin. 26811476 7 23 theme glycoproteins 1285:1297 arg1 sequence 1223:1230 the sequence 1219:1230 the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1219:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 2 24 theme kidney 365:370 arg1 balance 384:390 kidney electrolyte balance 365:390 kidney electrolyte balance 365:390 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 6 25 theme ZP-N 1042:1045 arg1 homodimerization 1054:1069 ZP-N domain homodimerization 1042:1069 ZP-N domain homodimerization 1042:1069 The structure of UMOD reveals that an extensive hydrophobic interface mediates ZP-N domain homodimerization. 26811476 4 26 theme homopolymers 667:678 arg1 importance 649:658 the functional importance 634:658 the functional importance of its homopolymers 634:678 Despite the functional importance of its homopolymers, no structural information is available on UMOD and how it self-assembles into filaments. 26811476 5 27 theme polymerization 812:825 arg1 regions 827:833 polymerization regions 812:833 polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 812:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 8 28 theme multidomain 1459:1469 arg1 proteins 1471:1478 structurally similar multidomain proteins 1438:1478 structurally similar multidomain proteins 1438:1478 Our data provide an example of how interdomain linker plasticity can modulate the function of structurally similar multidomain proteins. 26811476 3 29 from aggregation 479:489 arg1 urine 516:520 the urine 512:520 the urine 512:520 Moreover, salt-dependent aggregation of UMOD filaments in the urine generates a soluble molecular net that captures uropathogenic bacteria and facilitates their clearance. 26811476 1 30 theme /Tamm-Horsfall 98:111 arg1 protein 154:160 the most abundant human urinary protein 122:160 the most abundant human urinary protein 122:160 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 1 30 theme /Tamm-Horsfall 98:111 arg1 protein 113:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 0 31 theme interdomain 13:23 arg1 linker 25:30 A structured interdomain linker 0:30 A structured interdomain linker 0:30 A structured interdomain linker directs self-polymerization of human uromodulin. 26811476 9 32 theme UMOD 1511:1514 arg1 architecture 1495:1506 the architecture 1491:1506 the architecture of UMOD 1491:1514 Moreover, the architecture of UMOD rationalizes numerous pathogenic mutations in both UMOD and TECTA genes. 26811476 8 33 theme similar 1451:1457 arg1 proteins 1471:1478 structurally similar multidomain proteins 1438:1478 structurally similar multidomain proteins 1438:1478 Our data provide an example of how interdomain linker plasticity can modulate the function of structurally similar multidomain proteins. 26811476 2 34 theme zona 281:284 arg1 pellucida 286:294 its bipartite zona pellucida module (ZP-N/ZP-C) 267:313 its bipartite zona pellucida module (ZP-N/ZP-C) 267:313 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 1 35 theme chronic 183:189 arg1 diseases 198:205 chronic kidney diseases 183:205 chronic kidney diseases 183:205 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 7 36 theme ZP-N/ZP-C 1154:1162 arg1 linker 1164:1169 an ordered ZP-N/ZP-C linker 1143:1169 an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1143:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 1 37 theme kidney 191:196 arg1 diseases 198:205 chronic kidney diseases 183:205 chronic kidney diseases 183:205 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 8 38 theme linker 1391:1396 arg1 plasticity 1398:1407 interdomain linker plasticity 1379:1407 interdomain linker plasticity 1379:1407 Our data provide an example of how interdomain linker plasticity can modulate the function of structurally similar multidomain proteins. 26811476 2 39 theme innate 396:401 arg1 immunity 403:410 innate immunity 396:410 innate immunity 396:410 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 4 40 theme structural 684:693 arg1 information 695:705 no structural information 681:705 no structural information 681:705 Despite the functional importance of its homopolymers, no structural information is available on UMOD and how it self-assembles into filaments. 26811476 8 41 theme proteins 1471:1478 arg1 function 1426:1433 the function 1422:1433 the function of structurally similar multidomain proteins 1422:1478 Our data provide an example of how interdomain linker plasticity can modulate the function of structurally similar multidomain proteins. 26811476 8 42 theme interdomain 1379:1389 arg1 plasticity 1398:1407 interdomain linker plasticity 1379:1407 interdomain linker plasticity 1379:1407 Our data provide an example of how interdomain linker plasticity can modulate the function of structurally similar multidomain proteins. 26811476 4 43 theme functional 638:647 arg1 importance 649:658 the functional importance 634:658 the functional importance of its homopolymers 634:678 Despite the functional importance of its homopolymers, no structural information is available on UMOD and how it self-assembles into filaments. 26811476 2 44 theme renal 440:444 arg1 stones 446:451 renal stones 440:451 renal stones 440:451 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 5 45 theme essential 867:875 arg1 UMOD 844:847 human UMOD 838:847 human UMOD 838:847 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 45 theme essential 867:875 arg1 protein 892:898 an essential sperm receptor protein 864:898 an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 864:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 45 theme essential 867:875 arg1 related 921:927 related 921:927 related 921:927 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 0 46 theme uromodulin 69:78 arg1 self-polymerization 40:58 self-polymerization 40:58 self-polymerization of human uromodulin 40:78 A structured interdomain linker directs self-polymerization of human uromodulin. 26811476 2 47 dep pellucida 286:294 arg1 ZP-N/ZP-C 304:312 ZP-N/ZP-C 304:312 ZP-N/ZP-C 304:312 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 2 47 dep pellucida 286:294 arg1 module 296:301 module 296:301 its bipartite zona pellucida module (ZP-N/ZP-C) 267:313 Via its bipartite zona pellucida module (ZP-N/ZP-C), UMOD forms extracellular filaments that regulate kidney electrolyte balance and innate immunity, as well as protect against renal stones. 26811476 5 48 theme crystal 790:796 arg1 structures 798:807 the crystal structures 786:807 the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 786:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 49 dep UMOD 932:935 arg1 heteropolymers 947:960 heteropolymers 947:960 heteropolymers 947:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 3 50 theme uropathogenic 570:582 arg1 bacteria 584:591 uropathogenic bacteria 570:591 uropathogenic bacteria 570:591 Moreover, salt-dependent aggregation of UMOD filaments in the urine generates a soluble molecular net that captures uropathogenic bacteria and facilitates their clearance. 26811476 7 51 theme disease-associated 1252:1269 arg1 glycoproteins 1285:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins 1235:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1235:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 51 theme disease-associated 1252:1269 arg1 glycoprotein 1322:1333 glycoprotein 2 1322:1335 glycoprotein 2 (GP2) 1322:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 51 theme disease-associated 1252:1269 arg1 α-tectorin 1299:1308 α-tectorin 1299:1308 α-tectorin (TECTA) 1299:1316 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 52 gly glycoproteins 1285:1297 arg1 glycoproteins 1285:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins 1235:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1235:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 52 gly glycoproteins 1285:1297 arg1 glycoprotein 1322:1333 glycoprotein 2 1322:1335 glycoprotein 2 (GP2) 1322:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 52 gly glycoproteins 1285:1297 arg1 α-tectorin 1299:1308 α-tectorin 1299:1308 α-tectorin (TECTA) 1299:1316 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 53 theme homopolymeric 1271:1283 arg1 glycoproteins 1285:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins 1235:1297 deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2) 1235:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 53 theme homopolymeric 1271:1283 arg1 glycoprotein 1322:1333 glycoprotein 2 1322:1335 glycoprotein 2 (GP2) 1322:1341 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 7 53 theme homopolymeric 1271:1283 arg1 α-tectorin 1299:1308 α-tectorin 1299:1308 α-tectorin (TECTA) 1299:1316 This arrangement is required for filament formation and is directed by an ordered ZP-N/ZP-C linker that is not observed in ZP2 but is conserved in the sequence of deafness/Crohn's disease-associated homopolymeric glycoproteins α-tectorin (TECTA) and glycoprotein 2 (GP2). 26811476 1 54 theme abundant 131:138 arg1 protein 154:160 the most abundant human urinary protein 122:160 the most abundant human urinary protein 122:160 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 1 54 theme abundant 131:138 arg1 protein 113:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 6 55 theme hydrophobic 1011:1021 arg1 interface 1023:1031 an extensive hydrophobic interface 998:1031 an extensive hydrophobic interface 998:1031 The structure of UMOD reveals that an extensive hydrophobic interface mediates ZP-N domain homodimerization. 26811476 5 56 theme sperm 877:881 arg1 UMOD 844:847 human UMOD 838:847 human UMOD 838:847 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 56 theme sperm 877:881 arg1 protein 892:898 an essential sperm receptor protein 864:898 an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 864:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 5 56 theme sperm 877:881 arg1 related 921:927 related 921:927 related 921:927 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 9 57 theme numerous 1529:1536 arg1 mutations 1549:1557 numerous pathogenic mutations 1529:1557 numerous pathogenic mutations 1529:1557 Moreover, the architecture of UMOD rationalizes numerous pathogenic mutations in both UMOD and TECTA genes. 26811476 1 58 theme human 140:144 arg1 protein 154:160 the most abundant human urinary protein 122:160 the most abundant human urinary protein 122:160 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 1 58 theme human 140:144 arg1 protein 113:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 5 59 theme ZP2 859:861 arg1 regions 827:833 polymerization regions 812:833 polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 812:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 6 60 theme extensive 1001:1009 arg1 interface 1023:1031 an extensive hydrophobic interface 998:1031 an extensive hydrophobic interface 998:1031 The structure of UMOD reveals that an extensive hydrophobic interface mediates ZP-N domain homodimerization. 26811476 0 61 theme human 63:67 arg1 uromodulin 69:78 human uromodulin 63:78 human uromodulin 63:78 A structured interdomain linker directs self-polymerization of human uromodulin. 26811476 5 62 theme regions 827:833 arg1 structures 798:807 the crystal structures 786:807 the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers 786:960 Here, we report the crystal structures of polymerization regions of human UMOD and mouse ZP2, an essential sperm receptor protein that is structurally related to UMOD but forms heteropolymers. 26811476 1 63 theme urinary 146:152 arg1 protein 154:160 the most abundant human urinary protein 122:160 the most abundant human urinary protein 122:160 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 1 63 theme urinary 146:152 arg1 protein 113:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein 81:119 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 26811476 1 64 theme promising 216:224 arg1 target 238:243 a promising therapeutic target 214:243 a promising therapeutic target for hypertension 214:260 Uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant human urinary protein, plays a key role in chronic kidney diseases and is a promising therapeutic target for hypertension. 8665956 2 0 theme PSA-PCI 414:420 arg1 complex 422:428 a PSA-PCI complex 412:428 a PSA-PCI complex 412:428 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 4 1 with interaction 632:642 arg1 PSA 655:657 PSA 655:657 PSA 655:657 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 4 1 with interaction 632:642 arg1 PCI 663:665 PCI 663:665 PCI 663:665 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 3 2 with Digestion 431:439 arg1 PSA 465:467 PSA 465:467 PSA 465:467 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 9 3 from pH 1683:1684 arg1 plasma 1760:1765 seminal plasma 1752:1765 seminal plasma 1752:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 5 4 theme nM 970:971 arg1 Kd 961:962 a Kd 959:962 a Kd of 28 nM 959:971 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 9 5 theme divalent 1703:1710 arg1 cations 1712:1718 divalent cations 1703:1718 divalent cations 1703:1718 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 0 6 theme C 108:108 arg1 inhibitor 110:118 protein C inhibitor 100:118 protein C inhibitor 100:118 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 1 7 theme human 293:297 arg1 plasma 307:312 human seminal plasma 293:312 human seminal plasma 293:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 5 8 theme dissociation 907:918 arg1 kd 930:931 kd 930:931 kd 930:931 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 5 8 theme dissociation 907:918 arg1 constant 920:927 an apparent dissociation constant 895:927 an apparent dissociation constant (kd) of 41 nM 895:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 7 9 theme Sg 1168:1169 arg1 bindings 1156:1163 The bindings 1152:1163 The bindings of Sg II to both iPr2P-PSA and PCI 1152:1198 The bindings of Sg II to both iPr2P-PSA and PCI were influenced by pH, ionic strength, heparin, dextran sulfate, and divalent cations, particularly by Zn2+. 8665956 9 10 from cations 1712:1718 arg1 plasma 1760:1765 seminal plasma 1752:1765 seminal plasma 1752:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 0 11 theme protein 100:106 arg1 inhibitor 110:118 protein C inhibitor 100:118 protein C inhibitor 100:118 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 1 12 theme seminal 299:305 arg1 plasma 307:312 human seminal plasma 293:312 human seminal plasma 293:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 8 13 with Treatment 1309:1317 arg1 heparinase 1333:1342 heparinase 1333:1342 heparinase 1333:1342 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 8 13 with Treatment 1309:1317 arg1 N-glycanase 1359:1369 N-glycanase 1359:1369 N-glycanase 1359:1369 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 8 13 with Treatment 1309:1317 arg1 heparitinase 1345:1356 heparitinase 1345:1356 heparitinase 1345:1356 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 8 13 with Treatment 1309:1317 arg1 sialidase 1402:1410 O-glycanase following sialidase 1380:1410 O-glycanase following sialidase 1380:1410 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 9 14 theme ionic 1687:1691 arg1 strength 1693:1700 ionic strength 1687:1700 ionic strength 1687:1700 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 5 15 theme diisopropylfluorophosphate 846:871 arg1 complex 835:841 the solid-phase complex 819:841 the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM 819:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 1 16 theme protein 230:236 arg1 antigen 160:166 prostate-specific antigen 142:166 prostate-specific antigen (PSA) 142:172 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 16 theme protein 230:236 arg1 inhibitor 240:248 protein C inhibitor 230:248 protein C inhibitor (PCI) 230:254 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 16 theme protein 230:236 arg1 substrates 187:196 its protein substrates 175:196 its protein substrates 175:196 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 16 theme protein 230:236 arg1 PCI 251:253 PCI 251:253 PCI 251:253 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 16 theme protein 230:236 arg1 semenogelin 199:209 semenogelin 199:209 semenogelin (Sg) 199:214 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 16 theme protein 230:236 arg1 components 279:288 components 279:288 components of human seminal plasma 279:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 16 theme protein 230:236 arg1 protease 132:139 The serine protease 121:139 The serine protease 121:139 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 17 theme plasma 307:312 arg1 antigen 160:166 prostate-specific antigen 142:166 prostate-specific antigen (PSA) 142:172 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 17 theme plasma 307:312 arg1 inhibitor 240:248 protein C inhibitor 230:248 protein C inhibitor (PCI) 230:254 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 17 theme plasma 307:312 arg1 substrates 187:196 its protein substrates 175:196 its protein substrates 175:196 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 17 theme plasma 307:312 arg1 semenogelin 199:209 semenogelin 199:209 semenogelin (Sg) 199:214 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 17 theme plasma 307:312 arg1 components 279:288 components 279:288 components of human seminal plasma 279:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 17 theme plasma 307:312 arg1 protease 132:139 The serine protease 121:139 The serine protease 121:139 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 9 18 theme Sg 1579:1580 arg1 degradation 1564:1574 the PSA-catalyzed degradation 1546:1574 the PSA-catalyzed degradation of Sg in seminal plasma 1546:1598 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 19 theme PSA-catalyzed 1550:1562 arg1 degradation 1564:1574 the PSA-catalyzed degradation 1546:1574 the PSA-catalyzed degradation of Sg in seminal plasma 1546:1598 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 5 20 with complex 835:841 arg1 kd 930:931 kd 930:931 kd 930:931 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 5 20 with complex 835:841 arg1 constant 920:927 an apparent dissociation constant 895:927 an apparent dissociation constant (kd) of 41 nM 895:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 8 21 theme Sg 1442:1443 arg1 binding 1431:1437 the binding 1427:1437 the binding of Sg II to iPr2P-PSA and PCI 1427:1467 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 9 22 from Sg 1513:1514 arg1 vesicles 1527:1534 seminal vesicles 1519:1534 seminal vesicles 1519:1534 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 5 23 theme PSA 886:888 arg1 complex 835:841 the solid-phase complex 819:841 the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM 819:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 6 24 theme Sg 989:990 arg1 binding 978:984 The binding 974:984 The binding of Sg II to iPr2P-PSA 974:1006 The binding of Sg II to iPr2P-PSA was not affected by PCI and that of Sg II to PCI was not affected by iPr2P-PSA, suggesting that Sg II forms a ternary complex with PSA and PCI. 8665956 2 25 theme Sg 391:392 arg1 degradation 358:368 the PSA-catalyzed degradation 340:368 the PSA-catalyzed degradation of insoluble coagula Sg I + II 340:399 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 1 26 theme C 238:238 arg1 antigen 160:166 prostate-specific antigen 142:166 prostate-specific antigen (PSA) 142:172 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 26 theme C 238:238 arg1 inhibitor 240:248 protein C inhibitor 230:248 protein C inhibitor (PCI) 230:254 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 26 theme C 238:238 arg1 substrates 187:196 its protein substrates 175:196 its protein substrates 175:196 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 26 theme C 238:238 arg1 PCI 251:253 PCI 251:253 PCI 251:253 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 26 theme C 238:238 arg1 semenogelin 199:209 semenogelin 199:209 semenogelin (Sg) 199:214 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 26 theme C 238:238 arg1 components 279:288 components 279:288 components of human seminal plasma 279:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 26 theme C 238:238 arg1 protease 132:139 The serine protease 121:139 The serine protease 121:139 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 2 27 dep Sg 391:392 arg1 II 398:399 II 398:399 II 398:399 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 2 27 dep Sg 391:392 arg1 I 394:394 I 394:394 I 394:394 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 4 28 theme Sg 647:648 arg1 interaction 632:642 the molecular interaction 618:642 the molecular interaction of Sg with PSA and PCI 618:665 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 9 29 theme PCI 1625:1627 arg1 binding 1614:1620 the binding 1610:1620 the binding of PCI and PSA to Sg 1610:1641 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 1 30 theme protein 179:185 arg1 antigen 160:166 prostate-specific antigen 142:166 prostate-specific antigen (PSA) 142:172 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 30 theme protein 179:185 arg1 inhibitor 240:248 protein C inhibitor 230:248 protein C inhibitor (PCI) 230:254 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 30 theme protein 179:185 arg1 substrates 187:196 its protein substrates 175:196 its protein substrates 175:196 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 30 theme protein 179:185 arg1 semenogelin 199:209 semenogelin 199:209 semenogelin (Sg) 199:214 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 30 theme protein 179:185 arg1 components 279:288 components 279:288 components of human seminal plasma 279:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 30 theme protein 179:185 arg1 protease 132:139 The serine protease 121:139 The serine protease 121:139 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 2 31 theme coagula 383:389 arg1 Sg 391:392 insoluble coagula Sg I + II 373:399 insoluble coagula Sg I + II 373:399 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 4 32 theme seminal 691:697 arg1 coagula 699:705 seminal coagula 691:705 seminal coagula 691:705 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 6 33 theme ternary 1118:1124 arg1 complex 1126:1132 a ternary complex 1116:1132 a ternary complex with PSA and PCI 1116:1149 The binding of Sg II to iPr2P-PSA was not affected by PCI and that of Sg II to PCI was not affected by iPr2P-PSA, suggesting that Sg II forms a ternary complex with PSA and PCI. 8665956 5 34 theme solid-phase 823:833 arg1 complex 835:841 the solid-phase complex 819:841 the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM 819:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 2 35 theme insoluble 373:381 arg1 Sg 391:392 insoluble coagula Sg I + II 373:399 insoluble coagula Sg I + II 373:399 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 7 36 theme divalent 1269:1276 arg1 cations 1278:1284 divalent cations 1269:1284 divalent cations 1269:1284 The bindings of Sg II to both iPr2P-PSA and PCI were influenced by pH, ionic strength, heparin, dextran sulfate, and divalent cations, particularly by Zn2+. 8665956 3 37 theme PSA-PCI 486:492 arg1 complex 494:500 PSA-PCI complex 486:500 PSA-PCI complex 486:500 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 9 38 theme PSA 1633:1635 arg1 binding 1614:1620 the binding 1610:1620 the binding of PCI and PSA to Sg 1610:1641 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 3 39 theme seminal 444:450 arg1 coagula 452:458 seminal coagula 444:458 seminal coagula 444:458 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 3 40 theme active 568:573 arg1 PCI 575:577 active PCI 568:577 active PCI binding to the coagula 568:600 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 4 41 gly glycosylated 749:760 arg1 Sg 740:741 Sg II 740:744 Sg II 740:744 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 2 42 theme PSA-catalyzed 344:356 arg1 degradation 358:368 the PSA-catalyzed degradation 340:368 the PSA-catalyzed degradation of insoluble coagula Sg I + II 340:399 PCI was found to inhibit the PSA-catalyzed degradation of insoluble coagula Sg I + II by forming a PSA-PCI complex. 8665956 5 43 theme apparent 898:905 arg1 kd 930:931 kd 930:931 kd 930:931 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 5 43 theme apparent 898:905 arg1 constant 920:927 an apparent dissociation constant 895:927 an apparent dissociation constant (kd) of 41 nM 895:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 9 44 theme seminal 1519:1525 arg1 vesicles 1527:1534 seminal vesicles 1519:1534 seminal vesicles 1519:1534 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 4 45 theme heterogeneous 767:779 arg1 moieties 794:801 heterogeneous carbohydrate moieties 767:801 heterogeneous carbohydrate moieties 767:801 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 4 46 theme soluble 712:718 arg1 form 720:723 a soluble form 710:723 a soluble form 710:723 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 4 46 theme soluble 712:718 arg1 Sg 680:681 Sg II 680:684 Sg II 680:684 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 4 47 theme molecular 622:630 arg1 interaction 632:642 the molecular interaction 618:642 the molecular interaction of Sg with PSA and PCI 618:665 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 0 48 theme molecular 43:51 arg1 interaction 53:63 its molecular interaction 39:63 its molecular interaction with prostate-specific antigen and protein C inhibitor 39:118 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 8 49 theme Sg 1322:1323 arg1 Treatment 1309:1317 Treatment 1309:1317 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase 1309:1410 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 9 50 theme heparin-like 1725:1736 arg1 substances 1738:1747 heparin-like substances 1725:1747 heparin-like substances in seminal plasma 1725:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 7 51 theme dextran 1248:1254 arg1 sulfate 1256:1262 dextran sulfate 1248:1262 dextran sulfate 1248:1262 The bindings of Sg II to both iPr2P-PSA and PCI were influenced by pH, ionic strength, heparin, dextran sulfate, and divalent cations, particularly by Zn2+. 8665956 5 52 theme nM 940:941 arg1 kd 930:931 kd 930:931 kd 930:931 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 5 52 theme nM 940:941 arg1 constant 920:927 an apparent dissociation constant 895:927 an apparent dissociation constant (kd) of 41 nM 895:941 Sg II bound to the solid-phase complex of diisopropylfluorophosphate (iPr2FP) and PSA with an apparent dissociation constant (kd) of 41 nM and to PCI with a Kd of 28 nM. 8665956 1 53 theme serine 125:130 arg1 antigen 160:166 prostate-specific antigen 142:166 prostate-specific antigen (PSA) 142:172 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 53 theme serine 125:130 arg1 inhibitor 240:248 protein C inhibitor 230:248 protein C inhibitor (PCI) 230:254 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 53 theme serine 125:130 arg1 substrates 187:196 its protein substrates 175:196 its protein substrates 175:196 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 53 theme serine 125:130 arg1 semenogelin 199:209 semenogelin 199:209 semenogelin (Sg) 199:214 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 53 theme serine 125:130 arg1 components 279:288 components 279:288 components of human seminal plasma 279:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 53 theme serine 125:130 arg1 protease 132:139 The serine protease 121:139 The serine protease 121:139 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 9 54 theme seminal 1752:1758 arg1 plasma 1760:1765 seminal plasma 1752:1765 seminal plasma 1752:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 55 theme several 1659:1665 arg1 substances 1738:1747 heparin-like substances 1725:1747 heparin-like substances in seminal plasma 1725:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 55 theme several 1659:1665 arg1 factors 1667:1673 several factors 1659:1673 several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma 1659:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 55 theme several 1659:1665 arg1 pH 1683:1684 pH 1683:1684 pH 1683:1684 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 55 theme several 1659:1665 arg1 strength 1693:1700 ionic strength 1687:1700 ionic strength 1687:1700 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 55 theme several 1659:1665 arg1 cations 1712:1718 divalent cations 1703:1718 divalent cations 1703:1718 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 9 56 from substances 1738:1747 arg1 plasma 1760:1765 seminal plasma 1752:1765 seminal plasma 1752:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 0 57 with Characterization 0:15 arg1 antigen 88:94 prostate-specific antigen 70:94 prostate-specific antigen 70:94 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 0 57 with Characterization 0:15 arg1 inhibitor 110:118 protein C inhibitor 100:118 protein C inhibitor 100:118 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 7 58 theme ionic 1223:1227 arg1 strength 1229:1236 ionic strength 1223:1236 ionic strength 1223:1236 The bindings of Sg II to both iPr2P-PSA and PCI were influenced by pH, ionic strength, heparin, dextran sulfate, and divalent cations, particularly by Zn2+. 8665956 9 59 from degradation 1564:1574 arg1 plasma 1593:1598 seminal plasma 1585:1598 seminal plasma 1585:1598 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 4 60 theme carbohydrate 781:792 arg1 moieties 794:801 heterogeneous carbohydrate moieties 767:801 heterogeneous carbohydrate moieties 767:801 To investigate the molecular interaction of Sg with PSA and PCI, we purified Sg II from seminal coagula as a soluble form and found that Sg II is glycosylated with heterogeneous carbohydrate moieties. 8665956 3 61 theme PCI 575:577 arg1 presence 556:563 the presence 552:563 the presence of active PCI binding to the coagula 552:600 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 0 62 with interaction 53:63 arg1 antigen 88:94 prostate-specific antigen 70:94 prostate-specific antigen 70:94 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 0 62 with interaction 53:63 arg1 inhibitor 110:118 protein C inhibitor 100:118 protein C inhibitor 100:118 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 9 63 from strength 1693:1700 arg1 plasma 1760:1765 seminal plasma 1752:1765 seminal plasma 1752:1765 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 8665956 8 64 theme following 1392:1400 arg1 sialidase 1402:1410 O-glycanase following sialidase 1380:1410 O-glycanase following sialidase 1380:1410 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 0 65 theme prostate-specific 70:86 arg1 antigen 88:94 prostate-specific antigen 70:94 prostate-specific antigen 70:94 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 3 66 theme soluble 526:532 arg1 phase 534:538 a soluble phase 524:538 a soluble phase 524:538 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 6 67 with complex 1126:1132 arg1 PSA 1139:1141 PSA 1139:1141 PSA 1139:1141 The binding of Sg II to iPr2P-PSA was not affected by PCI and that of Sg II to PCI was not affected by iPr2P-PSA, suggesting that Sg II forms a ternary complex with PSA and PCI. 8665956 6 67 with complex 1126:1132 arg1 PCI 1147:1149 PCI 1147:1149 PCI 1147:1149 The binding of Sg II to iPr2P-PSA was not affected by PCI and that of Sg II to PCI was not affected by iPr2P-PSA, suggesting that Sg II forms a ternary complex with PSA and PCI. 8665956 1 68 theme prostate-specific 142:158 arg1 antigen 160:166 prostate-specific antigen 142:166 prostate-specific antigen (PSA) 142:172 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 68 theme prostate-specific 142:158 arg1 inhibitor 240:248 protein C inhibitor 230:248 protein C inhibitor (PCI) 230:254 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 68 theme prostate-specific 142:158 arg1 substrates 187:196 its protein substrates 175:196 its protein substrates 175:196 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 68 theme prostate-specific 142:158 arg1 PSA 169:171 PSA 169:171 PSA 169:171 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 68 theme prostate-specific 142:158 arg1 semenogelin 199:209 semenogelin 199:209 semenogelin (Sg) 199:214 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 68 theme prostate-specific 142:158 arg1 components 279:288 components 279:288 components of human seminal plasma 279:312 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 1 68 theme prostate-specific 142:158 arg1 protease 132:139 The serine protease 121:139 The serine protease 121:139 The serine protease, prostate-specific antigen (PSA), its protein substrates, semenogelin (Sg) I and II, and protein C inhibitor (PCI) have been described as components of human seminal plasma. 8665956 8 69 theme O-glycanase 1380:1390 arg1 sialidase 1402:1410 O-glycanase following sialidase 1380:1410 O-glycanase following sialidase 1380:1410 Treatment of Sg II with heparinase, heparitinase, N-glycanase, or with O-glycanase following sialidase did not affect the binding of Sg II to iPr2P-PSA and PCI. 8665956 0 70 theme semenogelin 20:30 arg1 Characterization 0:15 Characterization 0:15 Characterization of semenogelin II 0:33 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 0 70 theme semenogelin 20:30 arg1 interaction 53:63 its molecular interaction 39:63 its molecular interaction with prostate-specific antigen and protein C inhibitor 39:118 Characterization of semenogelin II and its molecular interaction with prostate-specific antigen and protein C inhibitor. 8665956 3 71 theme coagula 452:458 arg1 Digestion 431:439 Digestion 431:439 Digestion of seminal coagula with PSA 431:467 Digestion of seminal coagula with PSA released PCI and PSA-PCI complex from the coagula into a soluble phase, suggesting the presence of active PCI binding to the coagula. 8665956 9 72 theme seminal 1585:1591 arg1 plasma 1593:1598 seminal plasma 1585:1598 seminal plasma 1585:1598 These findings suggested that PCI bound to Sg in seminal vesicles regulates the PSA-catalyzed degradation of Sg in seminal plasma, and that the binding of PCI and PSA to Sg is modulated by several factors such as pH, ionic strength, divalent cations, and heparin-like substances in seminal plasma. 22547800 6 0 theme hyperpolarizing 1191:1205 arg1 direction 1207:1215 the hyperpolarizing direction 1187:1215 the hyperpolarizing direction 1187:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 7 1 theme distinct 1316:1323 arg1 expression 1325:1334 distinct expression 1316:1334 distinct expression in different human tissues 1316:1361 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 7 2 theme different 1339:1347 arg1 tissues 1355:1361 different human tissues 1339:1361 different human tissues 1339:1361 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 7 3 from LRRC26 1364:1369 arg1 glands 1402:1407 secretory glands 1392:1407 secretory glands 1392:1407 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 3 4 theme tissue-specific 649:663 arg1 β-subunits 675:684 the tissue-specific auxiliary β-subunits 645:684 the tissue-specific auxiliary β-subunits (β1-β4) 645:692 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 4 theme tissue-specific 649:663 arg1 β1-β4 687:691 β1-β4 687:691 β1-β4 687:691 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 8 5 theme different 1688:1696 arg1 tissues 1698:1704 different tissues 1688:1704 different tissues 1688:1704 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 4 6 theme -containing 745:755 arg1 protein 766:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 6 theme -containing 745:755 arg1 subunit 809:815 a BK channel auxiliary subunit 786:815 a BK channel auxiliary subunit 786:815 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 6 theme -containing 745:755 arg1 LRRC26 775:780 LRRC26 775:780 LRRC26 775:780 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 8 7 theme tissues 1698:1704 arg1 spectrum 1676:1683 a spectrum 1674:1683 a spectrum of different tissues or cell types 1674:1718 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 3 8 theme voltage- 386:393 arg1 channels 427:434 voltage- and calcium-activated potassium channels 386:434 voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) 386:449 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 9 theme auxiliary 665:673 arg1 β-subunits 675:684 the tissue-specific auxiliary β-subunits 645:684 the tissue-specific auxiliary β-subunits (β1-β4) 645:692 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 9 theme auxiliary 665:673 arg1 β1-β4 687:691 β1-β4 687:691 β1-β4 687:691 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 10 dep channels 427:434 arg1 K 441:441 K(Ca)1.1 441:448 K(Ca)1.1 441:448 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 10 dep channels 427:434 arg1 BK 437:438 BK 437:438 BK 437:438 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 5 11 theme paralogous 957:966 arg1 LRRC55 986:991 LRRC55 986:991 LRRC55 986:991 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 11 theme paralogous 957:966 arg1 LRRC38 998:1003 LRRC38 998:1003 LRRC38 998:1003 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 11 theme paralogous 957:966 arg1 LRRC52 978:983 LRRC52 978:983 LRRC52 978:983 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 11 theme paralogous 957:966 arg1 proteins 968:975 LRRC26 paralogous proteins 950:975 LRRC26 paralogous proteins 950:975 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 8 12 from β-subunits 1525:1534 arg1 distinct 1507:1514 distinct 1507:1514 distinct 1507:1514 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 2 13 theme ion 335:337 arg1 function 347:354 ion channel function 335:354 ion channel function 335:354 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 7 14 theme secretory 1392:1400 arg1 glands 1402:1407 secretory glands 1392:1407 secretory glands 1392:1407 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 1 15 theme Molecular 76:84 arg1 diversity 86:94 Molecular diversity 76:94 Molecular diversity of ion channel structure and function 76:132 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 6 16 theme activation 1173:1182 arg1 dependence 1159:1168 the BK channel's voltage dependence 1134:1168 the BK channel's voltage dependence of activation in the hyperpolarizing direction 1134:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 2 17 theme tissue- 297:303 arg1 diversity 322:330 tissue- or cell-specific diversity 297:330 tissue- or cell-specific diversity of ion channel function 297:354 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 6 18 from dependence 1159:1168 arg1 direction 1207:1215 the hyperpolarizing direction 1187:1215 the hyperpolarizing direction 1187:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 8 19 theme proteins 1600:1607 arg1 them 1553:1556 them 1553:1556 them 1553:1556 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 8 19 theme proteins 1600:1607 arg1 family 1565:1570 a γ family 1561:1570 a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types 1561:1718 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 4 20 theme channel 791:797 arg1 protein 766:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 20 theme channel 791:797 arg1 subunit 809:815 a BK channel auxiliary subunit 786:815 a BK channel auxiliary subunit 786:815 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 1 21 from signaling 170:178 arg1 cells 215:219 nonexcitable cells 202:219 nonexcitable cells 202:219 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 1 21 from signaling 170:178 arg1 muscle 190:195 muscle 190:195 muscle 190:195 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 1 21 from signaling 170:178 arg1 nerve 183:187 nerve 183:187 nerve 183:187 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 6 22 theme marked 1118:1123 arg1 shift 1125:1129 a marked shift 1116:1129 a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction 1116:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 4 23 theme BK 788:789 arg1 protein 766:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 23 theme BK 788:789 arg1 subunit 809:815 a BK channel auxiliary subunit 786:815 a BK channel auxiliary subunit 786:815 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 1 24 from variability 144:154 arg1 signaling 170:178 electrical signaling 159:178 electrical signaling in nerve, muscle, and nonexcitable cells 159:219 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 6 25 from activation 1173:1182 arg1 direction 1207:1215 the hyperpolarizing direction 1187:1215 the hyperpolarizing direction 1187:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 8 26 theme γ 1563:1563 arg1 them 1553:1556 them 1553:1556 them 1553:1556 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 8 26 theme γ 1563:1563 arg1 family 1565:1570 a γ family 1561:1570 a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types 1561:1718 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 0 27 theme potassium 3:11 arg1 modulation 21:30 BK potassium channel modulation 0:30 BK potassium channel modulation by leucine-rich repeat-containing proteins. 0:74 BK potassium channel modulation by leucine-rich repeat-containing proteins. 22547800 5 28 theme LRRC26-type 1034:1044 arg1 subunits 1056:1063 LRRC26-type auxiliary subunits 1034:1063 LRRC26-type auxiliary subunits of BK channels 1034:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 28 theme LRRC26-type 1034:1044 arg1 group 941:945 a group 939:945 a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels 939:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 28 theme LRRC26-type 1034:1044 arg1 proteins 968:975 LRRC26 paralogous proteins 950:975 LRRC26 paralogous proteins 950:975 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 6 29 theme voltage 1151:1157 arg1 dependence 1159:1168 the BK channel's voltage dependence 1134:1168 the BK channel's voltage dependence of activation in the hyperpolarizing direction 1134:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 3 30 from functions 491:499 arg1 tissues 514:520 different tissues 504:520 different tissues 504:520 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 30 from functions 491:499 arg1 types 530:534 cell types 525:534 cell types 525:534 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 1 31 theme ion 99:101 arg1 structure 111:119 ion channel structure 99:119 ion channel structure 99:119 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 7 32 from LRRC55 1432:1437 arg1 brain 1442:1446 brain 1442:1446 brain 1442:1446 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 7 33 dep show 1311:1314 arg1 LRRC38 1375:1380 LRRC38 1375:1380 LRRC38 1375:1380 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 7 33 dep show 1311:1314 arg1 LRRC55 1432:1437 LRRC55 1432:1437 LRRC55 1432:1437 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 7 33 dep show 1311:1314 arg1 LRRC52 1410:1415 LRRC52 1410:1415 LRRC52 1410:1415 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 7 33 dep show 1311:1314 arg1 LRRC26 1364:1369 LRRC26 1364:1369 LRRC26 1364:1369 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 0 34 theme BK 0:1 arg1 modulation 21:30 BK potassium channel modulation 0:30 BK potassium channel modulation by leucine-rich repeat-containing proteins. 0:74 BK potassium channel modulation by leucine-rich repeat-containing proteins. 22547800 4 35 theme auxiliary 799:807 arg1 protein 766:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 35 theme auxiliary 799:807 arg1 subunit 809:815 a BK channel auxiliary subunit 786:815 a BK channel auxiliary subunit 786:815 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 7 36 from expression 1325:1334 arg1 tissues 1355:1361 different human tissues 1339:1361 different human tissues 1339:1361 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 1 37 theme channel 103:109 arg1 structure 111:119 ion channel structure 99:119 ion channel structure 99:119 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 3 38 theme calcium-activated 399:415 arg1 channels 427:434 voltage- and calcium-activated potassium channels 386:434 voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) 386:449 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 39 dep expressed 468:476 arg1 either 615:620 either 615:620 either 615:620 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 39 dep expressed 468:476 arg1 β-subunits 675:684 the tissue-specific auxiliary β-subunits 645:684 the tissue-specific auxiliary β-subunits (β1-β4) 645:692 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 39 dep expressed 468:476 arg1 β1-β4 687:691 β1-β4 687:691 β1-β4 687:691 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 7 40 from LRRC52 1410:1415 arg1 testis 1420:1425 testis 1420:1425 testis 1420:1425 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 8 41 theme gating 1651:1656 arg1 properties 1658:1667 the channel's gating properties 1637:1667 the channel's gating properties 1637:1667 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 1 42 theme structure 111:119 arg1 diversity 86:94 Molecular diversity 76:94 Molecular diversity of ion channel structure and function 76:132 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 1 43 theme nonexcitable 202:213 arg1 cells 215:219 nonexcitable cells 202:219 nonexcitable cells 202:219 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 0 44 theme channel 13:19 arg1 modulation 21:30 BK potassium channel modulation 0:30 BK potassium channel modulation by leucine-rich repeat-containing proteins. 0:74 BK potassium channel modulation by leucine-rich repeat-containing proteins. 22547800 2 45 theme cell-specific 308:320 arg1 diversity 322:330 tissue- or cell-specific diversity 297:330 tissue- or cell-specific diversity of ion channel function 297:354 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 8 46 theme BK 1579:1580 arg1 proteins 1600:1607 the BK channel auxiliary proteins 1575:1607 the BK channel auxiliary proteins 1575:1607 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 3 47 theme cell 525:528 arg1 types 530:534 cell types 525:534 cell types 525:534 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 0 48 theme leucine-rich 35:46 arg1 proteins 66:73 leucine-rich repeat-containing proteins 35:73 leucine-rich repeat-containing proteins 35:73 BK potassium channel modulation by leucine-rich repeat-containing proteins. 22547800 8 49 theme cell 1709:1712 arg1 types 1714:1718 cell types 1709:1718 cell types 1709:1718 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 5 50 theme BK 1068:1069 arg1 channels 1071:1078 BK channels 1068:1078 BK channels 1068:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 6 51 theme calcium 1279:1285 arg1 absence 1268:1274 the absence 1264:1274 the absence of calcium 1264:1285 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 5 52 theme channels 1071:1078 arg1 subunits 1056:1063 LRRC26-type auxiliary subunits 1034:1063 LRRC26-type auxiliary subunits of BK channels 1034:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 52 theme channels 1071:1078 arg1 group 941:945 a group 939:945 a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels 939:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 52 theme channels 1071:1078 arg1 proteins 968:975 LRRC26 paralogous proteins 950:975 LRRC26 paralogous proteins 950:975 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 2 53 theme auxiliary 245:253 arg1 subunits 255:262 variable auxiliary subunits 236:262 variable auxiliary subunits 236:262 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 8 54 theme auxiliary 1590:1598 arg1 proteins 1600:1607 the BK channel auxiliary proteins 1575:1607 the BK channel auxiliary proteins 1575:1607 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 4 55 theme leucine-rich 720:731 arg1 LRR 741:743 LRR 741:743 LRR 741:743 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 55 theme leucine-rich 720:731 arg1 repeat 733:738 a leucine-rich repeat 718:738 a leucine-rich repeat (LRR) 718:744 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 56 theme voltage 882:888 arg1 dependence 890:899 voltage dependence 882:899 voltage dependence of channel activation 882:921 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 57 theme large 848:852 arg1 shift 863:867 an unprecedented large negative shift 831:867 an unprecedented large negative shift (∼140 mV) 831:877 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 57 theme large 848:852 arg1 mV 875:876 ∼140 mV 870:876 ∼140 mV 870:876 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 58 theme channel 904:910 arg1 activation 912:921 channel activation 904:921 channel activation 904:921 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 2 59 theme variable 236:243 arg1 subunits 255:262 variable auxiliary subunits 236:262 variable auxiliary subunits 236:262 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 8 60 theme channel 1582:1588 arg1 proteins 1600:1607 the BK channel auxiliary proteins 1575:1607 the BK channel auxiliary proteins 1575:1607 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 1 61 theme function 125:132 arg1 diversity 86:94 Molecular diversity 76:94 Molecular diversity of ion channel structure and function 76:132 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 0 62 theme repeat-containing 48:64 arg1 proteins 66:73 leucine-rich repeat-containing proteins 35:73 leucine-rich repeat-containing proteins 35:73 BK potassium channel modulation by leucine-rich repeat-containing proteins. 22547800 4 63 theme unprecedented 834:846 arg1 shift 863:867 an unprecedented large negative shift 831:867 an unprecedented large negative shift (∼140 mV) 831:877 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 63 theme unprecedented 834:846 arg1 mV 875:876 ∼140 mV 870:876 ∼140 mV 870:876 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 5 64 theme proteins 968:975 arg1 subunits 1056:1063 LRRC26-type auxiliary subunits 1034:1063 LRRC26-type auxiliary subunits of BK channels 1034:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 64 theme proteins 968:975 arg1 group 941:945 a group 939:945 a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels 939:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 64 theme proteins 968:975 arg1 LRRC52 978:983 LRRC52 978:983 LRRC52 978:983 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 64 theme proteins 968:975 arg1 LRRC55 986:991 LRRC55 986:991 LRRC55 986:991 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 64 theme proteins 968:975 arg1 proteins 968:975 LRRC26 paralogous proteins 950:975 LRRC26 paralogous proteins 950:975 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 64 theme proteins 968:975 arg1 LRRC38 998:1003 LRRC38 998:1003 LRRC38 998:1003 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 3 65 theme different 504:512 arg1 tissues 514:520 different tissues 504:520 different tissues 504:520 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 7 66 theme human 1349:1353 arg1 tissues 1355:1361 different human tissues 1339:1361 different human tissues 1339:1361 They along with LRRC26 show distinct expression in different human tissues: LRRC26 and LRRC38 mainly in secretory glands, LRRC52 in testis, and LRRC55 in brain. 22547800 3 67 theme voltage- 569:576 arg1 α-subunits 597:606 the pore-forming, voltage- and Ca(2+)-sensing α-subunits 551:606 α-subunits 597:606 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 67 theme voltage- 569:576 arg1 BKα 609:611 BKα 609:611 BKα 609:611 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 4 68 theme membrane 757:764 arg1 protein 766:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 a leucine-rich repeat (LRR)-containing membrane protein 718:772 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 68 theme membrane 757:764 arg1 subunit 809:815 a BK channel auxiliary subunit 786:815 a BK channel auxiliary subunit 786:815 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 68 theme membrane 757:764 arg1 LRRC26 775:780 LRRC26 775:780 LRRC26 775:780 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 6 69 theme BK 1138:1139 arg1 channel 1141:1147 the BK channel 1134:1147 the BK channel's voltage dependence of activation in the hyperpolarizing direction 1134:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 22547800 2 70 theme function 347:354 arg1 diversity 322:330 tissue- or cell-specific diversity 297:330 tissue- or cell-specific diversity of ion channel function 297:354 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 3 71 theme pore-forming 555:566 arg1 α-subunits 597:606 the pore-forming, voltage- and Ca(2+)-sensing α-subunits 551:606 α-subunits 597:606 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 71 theme pore-forming 555:566 arg1 BKα 609:611 BKα 609:611 BKα 609:611 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 4 72 theme negative 854:861 arg1 shift 863:867 an unprecedented large negative shift 831:867 an unprecedented large negative shift (∼140 mV) 831:877 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 4 72 theme negative 854:861 arg1 mV 875:876 ∼140 mV 870:876 ∼140 mV 870:876 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 2 73 theme major 269:273 arg1 Regulation 222:231 Regulation 222:231 Regulation by variable auxiliary subunits 222:262 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 2 73 theme major 269:273 arg1 mechanism 275:283 a major mechanism 267:283 a major mechanism to generate tissue- or cell-specific diversity of ion channel function 267:354 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 2 74 theme channel 339:345 arg1 function 347:354 ion channel function 335:354 ion channel function 335:354 Regulation by variable auxiliary subunits is a major mechanism to generate tissue- or cell-specific diversity of ion channel function. 22547800 3 75 theme diverse 483:489 arg1 functions 491:499 diverse functions 483:499 diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα) 483:612 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 4 76 theme activation 912:921 arg1 dependence 890:899 voltage dependence 882:899 voltage dependence of channel activation 882:921 We recently identified a leucine-rich repeat (LRR)-containing membrane protein, LRRC26, as a BK channel auxiliary subunit, which causes an unprecedented large negative shift (∼140 mV) in voltage dependence of channel activation. 22547800 3 77 theme -sensing 588:595 arg1 α-subunits 597:606 the pore-forming, voltage- and Ca(2+)-sensing α-subunits 551:606 α-subunits 597:606 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 77 theme -sensing 588:595 arg1 BKα 609:611 BKα 609:611 BKα 609:611 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 1 78 theme electrical 159:168 arg1 signaling 170:178 electrical signaling 159:178 electrical signaling in nerve, muscle, and nonexcitable cells 159:219 Molecular diversity of ion channel structure and function underlies variability in electrical signaling in nerve, muscle, and nonexcitable cells. 22547800 3 79 dep either 615:620 arg1 alone 622:626 alone 622:626 alone 622:626 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 3 80 theme Mammalian 357:365 arg1 large-conductance 367:383 Mammalian large-conductance 357:383 Mammalian large-conductance 357:383 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 5 81 theme auxiliary 1046:1054 arg1 subunits 1056:1063 LRRC26-type auxiliary subunits 1034:1063 LRRC26-type auxiliary subunits of BK channels 1034:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 81 theme auxiliary 1046:1054 arg1 group 941:945 a group 939:945 a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels 939:1078 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 5 81 theme auxiliary 1046:1054 arg1 proteins 968:975 LRRC26 paralogous proteins 950:975 LRRC26 paralogous proteins 950:975 Here we report a group of LRRC26 paralogous proteins, LRRC52, LRRC55, and LRRC38 that potentially function as LRRC26-type auxiliary subunits of BK channels. 22547800 3 82 theme potassium 417:425 arg1 channels 427:434 voltage- and calcium-activated potassium channels 386:434 voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) 386:449 Mammalian large-conductance, voltage- and calcium-activated potassium channels (BK, K(Ca)1.1) are ubiquitously expressed with diverse functions in different tissues or cell types, consisting of the pore-forming, voltage- and Ca(2+)-sensing α-subunits (BKα), either alone or together with the tissue-specific auxiliary β-subunits (β1-β4). 22547800 8 83 theme types 1714:1718 arg1 spectrum 1676:1683 a spectrum 1674:1683 a spectrum of different tissues or cell types 1674:1718 LRRC26 and its paralogs are structurally and functionally distinct from the β-subunits and we designate them as a γ family of the BK channel auxiliary proteins, which potentially regulate the channel's gating properties over a spectrum of different tissues or cell types. 22547800 6 84 from direction 1207:1215 arg1 dependence 1159:1168 the BK channel's voltage dependence 1134:1168 the BK channel's voltage dependence of activation in the hyperpolarizing direction 1134:1215 LRRC52, LRRC55, and LRRC38 produce a marked shift in the BK channel's voltage dependence of activation in the hyperpolarizing direction by ∼100 mV, 50 mV, and 20 mV, respectively, in the absence of calcium. 16169851 5 0 theme CD28-like 650:658 arg1 receptors 660:668 other CD28-like receptors 644:668 other CD28-like receptors 644:668 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 7 1 theme scattering 827:836 arg1 analysis 838:845 Light scattering analysis 821:845 Light scattering analysis 821:845 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 0 2 theme complex 72:78 arg1 structure 45:53 the crystal structure 33:53 the crystal structure of the BTLA-HVEM complex 33:78 Attenuating lymphocyte activity: the crystal structure of the BTLA-HVEM complex. 16169851 4 3 theme crystal 468:474 arg1 structure 476:484 the 2.8-A crystal structure 458:484 the 2.8-A crystal structure of the BTLA-HVEM complex 458:509 To better understand this interaction, we determined the 2.8-A crystal structure of the BTLA-HVEM complex. 16169851 5 4 theme unique 607:612 arg1 surface 622:628 a unique binding surface 605:628 a unique binding surface 605:628 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 6 5 theme virus 761:765 arg1 glycoprotein 767:778 herpes virus glycoprotein 754:778 herpes virus glycoprotein D 754:780 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 4 6 theme 2.8-A 462:466 arg1 structure 476:484 the 2.8-A crystal structure 458:484 the 2.8-A crystal structure of the BTLA-HVEM complex 458:509 To better understand this interaction, we determined the 2.8-A crystal structure of the BTLA-HVEM complex. 16169851 5 7 theme binding 614:620 arg1 surface 622:628 a unique binding surface 605:628 a unique binding surface 605:628 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 5 8 theme N-terminal 553:562 arg1 HVEM 588:591 HVEM 588:591 HVEM 588:591 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 5 8 theme N-terminal 553:562 arg1 domain 578:583 the N-terminal cysteine-rich domain 549:583 the N-terminal cysteine-rich domain of HVEM 549:591 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 3 9 theme tumor 319:323 arg1 superfamily 350:360 tumor necrosis factor receptor superfamily 319:360 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 5 10 theme cysteine-rich 564:576 arg1 HVEM 588:591 HVEM 588:591 HVEM 588:591 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 5 10 theme cysteine-rich 564:576 arg1 domain 578:583 the N-terminal cysteine-rich domain 549:583 the N-terminal cysteine-rich domain of HVEM 549:591 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 9 11 theme I-set 1084:1088 arg1 fold 1090:1093 an immunoglobulin I-set fold 1066:1093 an immunoglobulin I-set fold 1066:1093 Finally, BTLA adopts an immunoglobulin I-set fold. 16169851 8 12 used used 995:998 arg2 mutagenesis 971:981 Alanine-scanning mutagenesis 954:981 Alanine-scanning mutagenesis of HVEM 954:989 Alanine-scanning mutagenesis of HVEM was used to further define critical binding residues. 16169851 5 13 theme HVEM 588:591 arg1 HVEM 588:591 HVEM 588:591 HVEM 588:591 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 5 13 theme HVEM 588:591 arg1 domain 578:583 the N-terminal cysteine-rich domain 549:583 the N-terminal cysteine-rich domain of HVEM 549:591 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 6 14 gly glycoprotein 767:778 arg1 glycoprotein 767:778 herpes virus glycoprotein 754:778 herpes virus glycoprotein D 754:780 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 0 15 theme lymphocyte 12:21 arg1 activity 23:30 lymphocyte activity 12:30 lymphocyte activity 12:30 Attenuating lymphocyte activity: the crystal structure of the BTLA-HVEM complex. 16169851 7 16 theme BTLA 893:896 arg1 monomeric 901:909 monomeric 901:909 monomeric 901:909 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 7 16 theme BTLA 893:896 arg1 BTLA 893:896 BTLA 893:896 BTLA 893:896 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 7 16 theme BTLA 893:896 arg1 domain 883:888 the extracellular domain 865:888 the extracellular domain of BTLA 865:896 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 3 17 theme virus 382:386 arg1 mediator 394:401 herpes virus entry mediator 375:401 herpes virus entry mediator 375:401 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 17 theme virus 382:386 arg1 HVEM 369:372 the tumor necrosis factor receptor superfamily member HVEM 315:372 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 18 theme superfamily 350:360 arg1 member 362:367 the tumor necrosis factor receptor superfamily member 315:367 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 7 19 theme extracellular 869:881 arg1 monomeric 901:909 monomeric 901:909 monomeric 901:909 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 7 19 theme extracellular 869:881 arg1 BTLA 893:896 BTLA 893:896 BTLA 893:896 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 7 19 theme extracellular 869:881 arg1 domain 883:888 the extracellular domain 865:888 the extracellular domain of BTLA 865:896 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 1 20 theme CD28-like 86:94 arg1 proteins 96:103 Five CD28-like proteins 81:103 Five CD28-like proteins 81:103 Five CD28-like proteins exert positive or negative effects on immune cells. 16169851 3 21 theme entry 388:392 arg1 mediator 394:401 herpes virus entry mediator 375:401 herpes virus entry mediator 375:401 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 21 theme entry 388:392 arg1 HVEM 369:372 the tumor necrosis factor receptor superfamily member HVEM 315:372 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 22 theme necrosis 325:332 arg1 superfamily 350:360 tumor necrosis factor receptor superfamily 319:360 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 4 23 theme complex 503:509 arg1 structure 476:484 the 2.8-A crystal structure 458:484 the 2.8-A crystal structure of the BTLA-HVEM complex 458:509 To better understand this interaction, we determined the 2.8-A crystal structure of the BTLA-HVEM complex. 16169851 8 24 theme critical 1018:1025 arg1 residues 1035:1042 critical binding residues 1018:1042 critical binding residues 1018:1042 Alanine-scanning mutagenesis of HVEM was used to further define critical binding residues. 16169851 3 25 theme factor 334:339 arg1 superfamily 350:360 tumor necrosis factor receptor superfamily 319:360 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 6 26 theme binding 806:812 arg1 motif 814:818 a similar binding motif 796:818 a similar binding motif 796:818 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 6 27 from surface 731:737 arg1 HVEM 742:745 HVEM 742:745 HVEM 742:745 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 0 28 theme crystal 37:43 arg1 structure 45:53 the crystal structure 33:53 the crystal structure of the BTLA-HVEM complex 33:78 Attenuating lymphocyte activity: the crystal structure of the BTLA-HVEM complex. 16169851 6 29 theme similar 798:804 arg1 motif 814:818 a similar binding motif 796:818 a similar binding motif 796:818 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 2 30 theme B7 220:221 arg1 family 223:228 the B7 family 216:228 the B7 family 216:228 Only four of these five receptors interact with members of the B7 family. 16169851 3 31 theme herpes 375:380 arg1 mediator 394:401 herpes virus entry mediator 375:401 herpes virus entry mediator 375:401 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 31 theme herpes 375:380 arg1 HVEM 369:372 the tumor necrosis factor receptor superfamily member HVEM 315:372 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 10 32 theme structural 1104:1113 arg1 similarities 1115:1126 structural similarities 1104:1126 structural similarities to other CD28-like members 1104:1153 Despite structural similarities to other CD28-like members, BTLA represents a unique co-receptor. 16169851 8 33 theme binding 1027:1033 arg1 residues 1035:1042 critical binding residues 1018:1042 critical binding residues 1018:1042 Alanine-scanning mutagenesis of HVEM was used to further define critical binding residues. 16169851 6 34 theme same 726:729 arg1 surface 731:737 the same surface 722:737 the same surface on HVEM 722:745 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 3 35 theme member 362:367 arg1 mediator 394:401 herpes virus entry mediator 375:401 herpes virus entry mediator 375:401 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 35 theme member 362:367 arg1 HVEM 369:372 the tumor necrosis factor receptor superfamily member HVEM 315:372 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 36 theme lymphocyte 262:271 arg1 attenuator 273:282 T lymphocyte attenuator 260:282 T lymphocyte attenuator 260:282 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 1 37 theme positive 111:118 arg1 effects 132:138 positive or negative effects 111:138 positive or negative effects 111:138 Five CD28-like proteins exert positive or negative effects on immune cells. 16169851 4 38 theme BTLA-HVEM 493:501 arg1 complex 503:509 the BTLA-HVEM complex 489:509 the BTLA-HVEM complex 489:509 To better understand this interaction, we determined the 2.8-A crystal structure of the BTLA-HVEM complex. 16169851 7 39 theme 1:1 941:943 arg1 complex 945:951 a 1:1 complex 939:951 a 1:1 complex 939:951 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 16169851 9 40 theme immunoglobulin 1069:1082 arg1 fold 1090:1093 an immunoglobulin I-set fold 1066:1093 an immunoglobulin I-set fold 1066:1093 Finally, BTLA adopts an immunoglobulin I-set fold. 16169851 8 41 theme HVEM 986:989 arg1 mutagenesis 971:981 Alanine-scanning mutagenesis 954:981 Alanine-scanning mutagenesis of HVEM 954:989 Alanine-scanning mutagenesis of HVEM was used to further define critical binding residues. 16169851 0 42 dep Attenuating 0:10 arg1 structure 45:53 the crystal structure 33:53 the crystal structure of the BTLA-HVEM complex 33:78 Attenuating lymphocyte activity: the crystal structure of the BTLA-HVEM complex. 16169851 5 43 theme other 644:648 arg1 receptors 660:668 other CD28-like receptors 644:668 other CD28-like receptors 644:668 This structure shows that BTLA binds the N-terminal cysteine-rich domain of HVEM and employs a unique binding surface compared with other CD28-like receptors. 16169851 6 44 theme herpes 754:759 arg1 glycoprotein 767:778 herpes virus glycoprotein 754:778 herpes virus glycoprotein D 754:780 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 1 45 theme negative 123:130 arg1 effects 132:138 positive or negative effects 111:138 positive or negative effects 111:138 Five CD28-like proteins exert positive or negative effects on immune cells. 16169851 10 46 theme CD28-like 1137:1145 arg1 members 1147:1153 other CD28-like members 1131:1153 other CD28-like members 1131:1153 Despite structural similarities to other CD28-like members, BTLA represents a unique co-receptor. 16169851 3 47 theme T 260:260 arg1 lymphocyte 262:271 T lymphocyte 260:271 T lymphocyte attenuator 260:282 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 2 48 theme receptors 181:189 arg1 receptors 181:189 these five receptors 170:189 these five receptors 170:189 Only four of these five receptors interact with members of the B7 family. 16169851 2 48 theme receptors 181:189 arg1 four 162:165 four 162:165 four 162:165 Only four of these five receptors interact with members of the B7 family. 16169851 1 49 theme immune 143:148 arg1 cells 150:154 immune cells 143:154 immune cells 143:154 Five CD28-like proteins exert positive or negative effects on immune cells. 16169851 10 50 theme unique 1174:1179 arg1 co-receptor 1181:1191 a unique co-receptor 1172:1191 a unique co-receptor 1172:1191 Despite structural similarities to other CD28-like members, BTLA represents a unique co-receptor. 16169851 10 51 theme other 1131:1135 arg1 members 1147:1153 other CD28-like members 1131:1153 other CD28-like members 1131:1153 Despite structural similarities to other CD28-like members, BTLA represents a unique co-receptor. 16169851 0 52 theme BTLA-HVEM 62:70 arg1 complex 72:78 the BTLA-HVEM complex 58:78 the BTLA-HVEM complex 58:78 Attenuating lymphocyte activity: the crystal structure of the BTLA-HVEM complex. 16169851 6 53 theme glycoprotein 767:778 arg1 D 780:780 herpes virus glycoprotein D 754:780 herpes virus glycoprotein D 754:780 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 6 53 theme glycoprotein 767:778 arg1 gD 750:751 gD 750:751 gD (herpes virus glycoprotein D) 750:781 Moreover, the structure shows that BTLA recognizes the same surface on HVEM as gD (herpes virus glycoprotein D) and utilizes a similar binding motif. 16169851 8 54 theme Alanine-scanning 954:969 arg1 mutagenesis 971:981 Alanine-scanning mutagenesis 954:981 Alanine-scanning mutagenesis of HVEM 954:989 Alanine-scanning mutagenesis of HVEM was used to further define critical binding residues. 16169851 3 55 dep BTLA 248:251 arg1 B 254:254 B 254:254 B 254:254 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 3 55 dep BTLA 248:251 arg1 attenuator 273:282 T lymphocyte attenuator 260:282 T lymphocyte attenuator 260:282 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 2 56 theme family 223:228 arg1 members 205:211 members 205:211 members of the B7 family 205:228 Only four of these five receptors interact with members of the B7 family. 16169851 3 57 theme receptor 341:348 arg1 superfamily 350:360 tumor necrosis factor receptor superfamily 319:360 the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator) 315:402 The exception is BTLA (B and T lymphocyte attenuator), which instead interacts with the tumor necrosis factor receptor superfamily member HVEM (herpes virus entry mediator). 16169851 7 58 theme Light 821:825 arg1 analysis 838:845 Light scattering analysis 821:845 Light scattering analysis 821:845 Light scattering analysis demonstrates that the extracellular domain of BTLA is monomeric and that BTLA and HVEM form a 1:1 complex. 17400922 7 0 theme free 1127:1130 arg1 ephrin-A5 1132:1140 free ephrin-A5 1127:1140 free ephrin-A5 1127:1140 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 5 1 theme key 845:847 arg1 topology 869:876 the Greek key beta-barrel folding topology 835:876 the Greek key beta-barrel folding topology 835:876 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 1 2 theme ephrin 134:139 arg1 mediators 163:171 important mediators 153:171 important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems 153:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 2 theme ephrin 134:139 arg1 receptors 61:69 The Eph receptors 53:69 The Eph receptors 53:69 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 2 theme ephrin 134:139 arg1 ligands 141:147 their ephrin ligands 128:147 their ephrin ligands 128:147 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 8 3 from rearrangement 1404:1416 arg1 ephrin-B2 1421:1429 ephrin-B2 1421:1429 ephrin-B2 1421:1429 Interestingly, the G-H loop undergoes a very similar conformational rearrangement in ephrin-B2 upon receptor binding. 17400922 9 4 theme starting 1592:1599 arg1 results 1458:1464 The results 1454:1464 The results of this study 1454:1478 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 9 4 theme starting 1592:1599 arg1 point 1601:1605 a starting point 1590:1605 a starting point for the development of structure-based Eph antagonists 1590:1660 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 5 5 theme disulphide 932:941 arg1 bonds 943:947 two disulphide bonds 928:947 two disulphide bonds 928:947 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 3 6 theme molecular 519:527 arg1 mechanism 529:537 the precise molecular mechanism 507:537 the precise molecular mechanism underlining their binding-partner preferences and subclass specificity 507:608 Nevertheless, open questions remain, including understanding the precise molecular mechanism underlining their binding-partner preferences and subclass specificity. 17400922 9 7 theme G-H 1520:1522 arg1 loop 1524:1527 the G-H loop 1516:1527 the G-H loop 1516:1527 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 7 8 theme significant 1218:1228 arg1 changes 1245:1251 significant conformational changes 1218:1251 significant conformational changes 1218:1251 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 9 9 theme antagonists 1650:1660 arg1 development 1615:1625 the development 1611:1625 the development of structure-based Eph antagonists 1611:1660 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 7 10 theme conformational 1230:1243 arg1 changes 1245:1251 significant conformational changes 1218:1251 significant conformational changes 1218:1251 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 5 11 contain containing 879:888 arg2 beta-strands 896:907 eight beta-strands 890:907 eight beta-strands 890:907 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 5 11 contain containing 879:888 arg1 topology 869:876 the Greek key beta-barrel folding topology 835:876 the Greek key beta-barrel folding topology 835:876 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 1 12 theme Eph 57:59 arg1 receptors 61:69 The Eph receptors 53:69 The Eph receptors 53:69 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 12 theme Eph 57:59 arg1 mediators 163:171 important mediators 153:171 important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems 153:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 12 theme Eph 57:59 arg1 subfamily 84:92 the largest subfamily 72:92 the largest subfamily of receptor tyrosine kinases 72:121 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 12 theme Eph 57:59 arg1 ligands 141:147 their ephrin ligands 128:147 their ephrin ligands 128:147 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 9 13 theme receptor 1533:1540 arg1 recognition 1542:1552 receptor recognition 1533:1552 receptor recognition 1533:1552 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 1 14 theme receptor 97:104 arg1 kinases 115:121 receptor tyrosine kinases 97:121 receptor tyrosine kinases 97:121 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 5 15 theme beta-barrel 849:859 arg1 topology 869:876 the Greek key beta-barrel folding topology 835:876 the Greek key beta-barrel folding topology 835:876 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 4 16 theme extracellular 686:698 arg1 structure 730:738 ephrin-A5-the first structure 710:738 ephrin-A5-the first structure of an unbound A-class ephrin 710:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 4 16 theme extracellular 686:698 arg1 domain 700:705 the extracellular domain 682:705 the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin 682:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 3 17 theme binding-partner 557:571 arg1 preferences 573:583 their binding-partner preferences 551:583 their binding-partner preferences 551:583 Nevertheless, open questions remain, including understanding the precise molecular mechanism underlining their binding-partner preferences and subclass specificity. 17400922 1 18 theme important 153:161 arg1 mediators 163:171 important mediators 153:171 important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems 153:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 18 theme important 153:161 arg1 receptors 61:69 The Eph receptors 53:69 The Eph receptors 53:69 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 18 theme important 153:161 arg1 ligands 141:147 their ephrin ligands 128:147 their ephrin ligands 128:147 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 7 19 theme EphB2-bound 1183:1193 arg1 ephrin-A5 1195:1203 EphB2-bound ephrin-A5 1183:1203 EphB2-bound ephrin-A5 1183:1203 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 8 20 theme conformational 1389:1402 arg1 rearrangement 1404:1416 a very similar conformational rearrangement 1374:1416 a very similar conformational rearrangement in ephrin-B2 1374:1429 Interestingly, the G-H loop undergoes a very similar conformational rearrangement in ephrin-B2 upon receptor binding. 17400922 9 21 theme structure-based 1630:1644 arg1 antagonists 1650:1660 structure-based Eph antagonists 1630:1660 structure-based Eph antagonists 1630:1660 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 9 22 theme loop 1524:1527 arg1 importance 1502:1511 the importance 1498:1511 the importance of the G-H loop for receptor recognition and selectivity 1498:1568 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 0 23 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human ephrin-A5 ectodomain 0:50 Crystal structure of the human ephrin-A5 ectodomain. 17400922 1 24 theme nervous 261:267 arg1 systems 288:294 the nervous and cardiovascular systems 257:294 the nervous and cardiovascular systems 257:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 25 theme largest 76:82 arg1 receptors 61:69 The Eph receptors 53:69 The Eph receptors 53:69 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 25 theme largest 76:82 arg1 subfamily 84:92 the largest subfamily 72:92 the largest subfamily of receptor tyrosine kinases 72:121 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 4 26 theme domain 700:705 arg1 structure 669:677 the crystal structure 657:677 the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin 657:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 2 27 theme ephrins 437:443 arg1 properties 414:423 the biochemical and signaling properties 384:423 the biochemical and signaling properties of Ephs and ephrins 384:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 7 28 theme ephrin 1279:1284 arg1 loop 1286:1289 the G-H ephrin loop 1271:1289 the G-H ephrin loop that upon binding bends toward the receptor 1271:1333 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 2 29 theme Recent 297:302 arg1 studies 315:321 Recent structural studies 297:321 Recent structural studies 297:321 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 2 30 theme Ephs 428:431 arg1 properties 414:423 the biochemical and signaling properties 384:423 the biochemical and signaling properties of Ephs and ephrins 384:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 2 31 theme structural 304:313 arg1 studies 315:321 Recent structural studies 297:321 Recent structural studies 297:321 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 1 32 theme cardiovascular 273:286 arg1 systems 288:294 the nervous and cardiovascular systems 257:294 the nervous and cardiovascular systems 257:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 4 33 theme structure 730:738 arg1 structure 730:738 ephrin-A5-the first structure 710:738 ephrin-A5-the first structure of an unbound A-class ephrin 710:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 4 33 theme structure 730:738 arg1 domain 700:705 the extracellular domain 682:705 the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin 682:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 1 34 theme communication 186:198 arg1 mediators 163:171 important mediators 153:171 important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems 153:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 34 theme communication 186:198 arg1 receptors 61:69 The Eph receptors 53:69 The Eph receptors 53:69 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 34 theme communication 186:198 arg1 ligands 141:147 their ephrin ligands 128:147 their ephrin ligands 128:147 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 0 35 theme ephrin-A5 31:39 arg1 ectodomain 41:50 the human ephrin-A5 ectodomain 21:50 the human ephrin-A5 ectodomain 21:50 Crystal structure of the human ephrin-A5 ectodomain. 17400922 2 36 theme unique 337:342 arg1 features 354:361 unique molecular features 337:361 unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins 337:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 5 37 theme folding 861:867 arg1 topology 869:876 the Greek key beta-barrel folding topology 835:876 the Greek key beta-barrel folding topology 835:876 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 2 38 theme properties 414:423 arg1 properties 414:423 the biochemical and signaling properties 384:423 the biochemical and signaling properties of Ephs and ephrins 384:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 2 38 theme properties 414:423 arg1 many 376:379 many 376:379 many 376:379 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 0 39 theme human 25:29 arg1 ectodomain 41:50 the human ephrin-A5 ectodomain 21:50 the human ephrin-A5 ectodomain 21:50 Crystal structure of the human ephrin-A5 ectodomain. 17400922 7 40 theme published 1160:1168 arg1 structure 1170:1178 the previously published structure 1145:1178 the previously published structure of EphB2-bound ephrin-A5 1145:1203 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 5 41 theme topology 869:876 arg1 variation 822:830 a variation 820:830 a variation of the Greek key beta-barrel folding topology, containing eight beta-strands 820:907 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 2 42 theme signaling 404:412 arg1 properties 414:423 the biochemical and signaling properties 384:423 the biochemical and signaling properties of Ephs and ephrins 384:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 3 43 theme precise 511:517 arg1 mechanism 529:537 the precise molecular mechanism 507:537 the precise molecular mechanism underlining their binding-partner preferences and subclass specificity 507:608 Nevertheless, open questions remain, including understanding the precise molecular mechanism underlining their binding-partner preferences and subclass specificity. 17400922 5 44 theme Greek 839:843 arg1 topology 869:876 the Greek key beta-barrel folding topology 835:876 the Greek key beta-barrel folding topology 835:876 The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. 17400922 8 45 theme similar 1381:1387 arg1 rearrangement 1404:1416 a very similar conformational rearrangement 1374:1416 a very similar conformational rearrangement in ephrin-B2 1374:1429 Interestingly, the G-H loop undergoes a very similar conformational rearrangement in ephrin-B2 upon receptor binding. 17400922 2 46 theme molecular 344:352 arg1 features 354:361 unique molecular features 337:361 unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins 337:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 0 47 theme ectodomain 41:50 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of the human ephrin-A5 ectodomain 0:50 Crystal structure of the human ephrin-A5 ectodomain. 17400922 7 48 theme G-H 1275:1277 arg1 loop 1286:1289 the G-H ephrin loop 1271:1289 the G-H ephrin loop that upon binding bends toward the receptor 1271:1333 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 17400922 8 49 theme receptor 1436:1443 arg1 binding 1445:1451 receptor binding 1436:1451 receptor binding 1436:1451 Interestingly, the G-H loop undergoes a very similar conformational rearrangement in ephrin-B2 upon receptor binding. 17400922 1 50 theme tyrosine 106:113 arg1 kinases 115:121 receptor tyrosine kinases 97:121 receptor tyrosine kinases 97:121 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 4 51 theme unbound 746:752 arg1 ephrin 762:767 an unbound A-class ephrin 743:767 an unbound A-class ephrin 743:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 9 52 theme Eph 1646:1648 arg1 antagonists 1650:1660 structure-based Eph antagonists 1630:1660 structure-based Eph antagonists 1630:1660 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 1 53 theme kinases 115:121 arg1 receptors 61:69 The Eph receptors 53:69 The Eph receptors 53:69 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 1 53 theme kinases 115:121 arg1 subfamily 84:92 the largest subfamily 72:92 the largest subfamily of receptor tyrosine kinases 72:121 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 3 54 theme subclass 589:596 arg1 specificity 598:608 subclass specificity 589:608 subclass specificity 589:608 Nevertheless, open questions remain, including understanding the precise molecular mechanism underlining their binding-partner preferences and subclass specificity. 17400922 9 55 theme study 1474:1478 arg1 point 1601:1605 a starting point 1590:1605 a starting point for the development of structure-based Eph antagonists 1590:1660 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 9 55 theme study 1474:1478 arg1 results 1458:1464 The results 1454:1464 The results of this study 1454:1478 The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. 17400922 1 56 dep cell 211:214 arg1 attachment 216:225 attachment 216:225 attachment 216:225 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 4 57 theme A-class 754:760 arg1 ephrin 762:767 an unbound A-class ephrin 743:767 an unbound A-class ephrin 743:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 8 58 theme G-H 1355:1357 arg1 loop 1359:1362 the G-H loop 1351:1362 the G-H loop 1351:1362 Interestingly, the G-H loop undergoes a very similar conformational rearrangement in ephrin-B2 upon receptor binding. 17400922 4 59 theme ephrin 762:767 arg1 structure 730:738 ephrin-A5-the first structure 710:738 ephrin-A5-the first structure of an unbound A-class ephrin 710:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 3 60 theme open 460:463 arg1 questions 465:473 open questions 460:473 open questions 460:473 Nevertheless, open questions remain, including understanding the precise molecular mechanism underlining their binding-partner preferences and subclass specificity. 17400922 1 61 theme cell-cell 176:184 arg1 communication 186:198 cell-cell communication 176:198 cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems 176:294 The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. 17400922 4 62 theme first 724:728 arg1 structure 730:738 ephrin-A5-the first structure 710:738 ephrin-A5-the first structure of an unbound A-class ephrin 710:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 2 63 theme biochemical 388:398 arg1 properties 414:423 the biochemical and signaling properties 384:423 the biochemical and signaling properties of Ephs and ephrins 384:443 Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. 17400922 4 64 theme crystal 661:667 arg1 structure 669:677 the crystal structure 657:677 the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin 657:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 4 65 theme ephrin-A5-the 710:722 arg1 structure 730:738 ephrin-A5-the first structure 710:738 ephrin-A5-the first structure of an unbound A-class ephrin 710:767 In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A-class ephrin. 17400922 7 66 theme ephrin-A5 1195:1203 arg1 structure 1170:1178 the previously published structure 1145:1178 the previously published structure of EphB2-bound ephrin-A5 1145:1203 Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. 21908432 10 0 with interface 1404:1412 arg1 GPIX 1419:1422 GPIX 1419:1422 GPIX 1419:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 3 1 with repeat 455:460 arg1 C-terminal 474:483 C-terminal 474:483 C-terminal 474:483 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 3 1 with repeat 455:460 arg1 N- 467:468 N- 467:468 N- 467:468 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 5 2 from tetramer 693:700 arg1 crystal 709:715 the crystal 705:715 the crystal 705:715 The chimera (GPIbβ(Eabc)), but not GPIbβ(E), forms a tetramer in the crystal, showing a quaternary interface between GPIbβ and GPIX. 21908432 1 3 theme GPIb-IX 157:163 arg1 complex 174:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 4 4 theme chimera 539:545 arg1 structure 524:532 The structure 520:532 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence 520:617 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 2 5 theme Bernard-Soulier 317:331 arg1 syndrome 333:340 Bernard-Soulier syndrome 317:340 Bernard-Soulier syndrome 317:340 Dysfunction in surface expression of the complex leads to Bernard-Soulier syndrome. 21908432 7 6 theme Mutagenesis 895:905 arg1 studies 907:913 Mutagenesis studies 895:913 Mutagenesis studies 895:913 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 8 7 with patients 1080:1087 arg1 syndrome 1110:1117 Bernard-Soulier syndrome 1094:1117 Bernard-Soulier syndrome 1094:1117 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 9 8 theme GPIX 1302:1305 arg1 expression 1315:1324 GPIX surface expression 1302:1324 GPIX surface expression 1302:1324 Two mutations, A108P and P74R, were found to maintain normal secretion/folding of GPIbβ(E) but were unable to support GPIX surface expression. 21908432 10 9 theme complex 1490:1496 arg1 organization 1462:1473 the quaternary organization 1447:1473 the quaternary organization of the GPIb-IX complex 1447:1496 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 3 10 theme capping 503:509 arg1 regions 511:517 disulphide-bonded capping regions 485:517 disulphide-bonded capping regions 485:517 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 4 11 theme GPIbβ 550:554 arg1 chimera 539:545 a chimera 537:545 a chimera of GPIbβ(E) and 3 loops (a,b,c) 537:577 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 9 12 theme surface 1307:1313 arg1 expression 1315:1324 GPIX surface expression 1302:1324 GPIX surface expression 1302:1324 Two mutations, A108P and P74R, were found to maintain normal secretion/folding of GPIbβ(E) but were unable to support GPIX surface expression. 21908432 0 13 theme GPIbβ 116:120 arg1 structures 102:111 the structures 98:111 the structures of GPIbβ and a GPIbβ/GPIX chimera 98:145 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 4 14 theme ectodomain 599:608 arg1 sequence 610:617 the GPIX ectodomain sequence 590:617 the GPIX ectodomain sequence 590:617 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 8 15 theme surface 1164:1170 arg1 expression 1172:1181 GPIb-IX complex surface expression 1148:1181 GPIb-IX complex surface expression 1148:1181 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 8 16 theme GPIbβ 1039:1043 arg1 mutations 1054:1062 Eight GPIbβ missense mutations 1033:1062 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome 1033:1117 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 8 17 theme complex 1156:1162 arg1 expression 1172:1181 GPIb-IX complex surface expression 1148:1181 GPIb-IX complex surface expression 1148:1181 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 3 18 theme GPIbβ 368:372 arg1 GPIbβ 386:390 GPIbβ 386:390 GPIbβ(E) 386:393 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 3 18 theme GPIbβ 368:372 arg1 ectodomain 374:383 the GPIbβ ectodomain 364:383 the GPIbβ ectodomain (GPIbβ(E)) 364:394 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 7 19 theme interactions 969:980 arg1 interface 930:938 this interface 925:938 this interface 925:938 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 7 19 theme interactions 969:980 arg1 representation 951:964 a valid representation 943:964 a valid representation of interactions between GPIbβ and GPIX in the full-length complex 943:1030 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 6 20 from GPIbβ 822:826 arg1 residue 802:808 residue Tyr106 802:815 residue Tyr106 802:815 Central to this interface is residue Tyr106 from GPIbβ, which inserts into a pocket generated by 2 loops (b,c) from GPIX. 21908432 7 21 from GPIbβ 990:994 arg1 complex 1024:1030 the full-length complex 1008:1030 the full-length complex 1008:1030 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 0 22 theme Quaternary 0:9 arg1 organization 11:22 Quaternary organization 0:22 Quaternary organization of GPIb-IX complex 0:41 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 0 23 theme GPIb-IX 27:33 arg1 complex 35:41 GPIb-IX complex 27:41 GPIb-IX complex 27:41 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 1 24 theme 1:2:1 252:256 arg1 ratio 243:247 a ratio 241:247 a ratio of 1:2:1 241:256 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 2 25 theme surface 274:280 arg1 expression 282:291 surface expression 274:291 surface expression of the complex 274:306 Dysfunction in surface expression of the complex leads to Bernard-Soulier syndrome. 21908432 4 26 theme loops 565:569 arg1 chimera 539:545 a chimera 537:545 a chimera of GPIbβ(E) and 3 loops (a,b,c) 537:577 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 7 27 theme valid 945:949 arg1 interface 930:938 this interface 925:938 this interface 925:938 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 7 27 theme valid 945:949 arg1 representation 951:964 a valid representation 943:964 a valid representation of interactions between GPIbβ and GPIX in the full-length complex 943:1030 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 1 28 theme receptor 165:172 arg1 complex 174:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 8 29 theme missense 1045:1052 arg1 mutations 1054:1062 Eight GPIbβ missense mutations 1033:1062 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome 1033:1117 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 4 30 theme GPIX 594:597 arg1 sequence 610:617 the GPIX ectodomain sequence 590:617 the GPIX ectodomain sequence 590:617 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 0 31 theme chimera 139:145 arg1 structures 102:111 the structures 98:111 the structures of GPIbβ and a GPIbβ/GPIX chimera 98:145 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 10 32 theme quaternary 1451:1460 arg1 organization 1462:1473 the quaternary organization 1447:1473 the quaternary organization of the GPIb-IX complex 1447:1496 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 0 33 theme complex 35:41 arg1 insights 47:54 insights 47:54 insights into Bernard-Soulier syndrome 47:84 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 0 33 theme complex 35:41 arg1 organization 11:22 Quaternary organization 0:22 Quaternary organization of GPIb-IX complex 0:41 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 0 34 theme GPIbβ/GPIX 128:137 arg1 chimera 139:145 a GPIbβ/GPIX chimera 126:145 a GPIbβ/GPIX chimera 126:145 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 1 35 contain has 182:184 arg1 complex 174:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 35 contain has 182:184 arg2 GPIbβ 204:208 GPIbβ 204:208 GPIbβ 204:208 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 35 contain has 182:184 arg2 subunits 188:195 3 subunits 186:195 3 subunits GPIbα, GPIbβ, and GPIX 186:218 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 35 contain has 182:184 arg2 GPIbα 197:201 GPIbα 197:201 GPIbα 197:201 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 35 contain has 182:184 arg2 GPIX 215:218 GPIX 215:218 GPIX 215:218 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 36 dep subunits 188:195 arg1 subunits 188:195 3 subunits 186:195 3 subunits GPIbα, GPIbβ, and GPIX 186:218 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 36 dep subunits 188:195 arg1 GPIbβ 204:208 GPIbβ 204:208 GPIbβ 204:208 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 36 dep subunits 188:195 arg1 GPIX 215:218 GPIX 215:218 GPIX 215:218 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 1 36 dep subunits 188:195 arg1 GPIbα 197:201 GPIbα 197:201 GPIbα 197:201 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 10 37 with Tyr106 1380:1385 arg1 GPIX 1419:1422 GPIX 1419:1422 GPIX 1419:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 6 38 dep b 879:879 arg1 c 881:881 c 881:881 c 881:881 Central to this interface is residue Tyr106 from GPIbβ, which inserts into a pocket generated by 2 loops (b,c) from GPIX. 21908432 10 39 theme mutations 1367:1375 arg1 proximity 1348:1356 The close structural proximity 1327:1356 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX 1327:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 7 40 theme full-length 1012:1022 arg1 complex 1024:1030 the full-length complex 1008:1030 the full-length complex 1008:1030 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 4 41 dep loops 565:569 arg1 b 574:574 b 574:574 b 574:574 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 4 41 dep loops 565:569 arg1 a 572:572 a 572:572 a 572:572 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 4 41 dep loops 565:569 arg1 c 576:576 c 576:576 c 576:576 The structure of a chimera of GPIbβ(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. 21908432 3 42 theme single 435:440 arg1 repeat 455:460 a single leucine-rich repeat 433:460 a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions 433:517 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 10 43 theme GPIbβ 1395:1399 arg1 interface 1404:1412 the GPIbβ(E) interface 1391:1412 the GPIbβ(E) interface with GPIX 1391:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 7 44 from GPIX 1000:1003 arg1 complex 1024:1030 the full-length complex 1008:1030 the full-length complex 1008:1030 Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIbβ and GPIX in the full-length complex. 21908432 3 45 theme leucine-rich 442:453 arg1 repeat 455:460 a single leucine-rich repeat 433:460 a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions 433:517 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 10 46 theme close 1331:1335 arg1 proximity 1348:1356 The close structural proximity 1327:1356 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX 1327:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 2 47 theme complex 300:306 arg1 expression 282:291 surface expression 274:291 surface expression of the complex 274:306 Dysfunction in surface expression of the complex leads to Bernard-Soulier syndrome. 21908432 3 48 dep N- 467:468 arg1 regions 511:517 disulphide-bonded capping regions 485:517 disulphide-bonded capping regions 485:517 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 3 49 theme disulphide-bonded 485:501 arg1 regions 511:517 disulphide-bonded capping regions 485:517 disulphide-bonded capping regions 485:517 We have crystallized the GPIbβ ectodomain (GPIbβ(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. 21908432 5 50 theme quaternary 728:737 arg1 interface 739:747 a quaternary interface 726:747 a quaternary interface between GPIbβ and GPIX 726:770 The chimera (GPIbβ(Eabc)), but not GPIbβ(E), forms a tetramer in the crystal, showing a quaternary interface between GPIbβ and GPIX. 21908432 8 51 theme GPIb-IX 1148:1154 arg1 expression 1172:1181 GPIb-IX complex surface expression 1148:1181 GPIb-IX complex surface expression 1148:1181 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 8 52 theme Bernard-Soulier 1094:1108 arg1 syndrome 1110:1117 Bernard-Soulier syndrome 1094:1117 Bernard-Soulier syndrome 1094:1117 Eight GPIbβ missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. 21908432 2 53 from Dysfunction 259:269 arg1 expression 282:291 surface expression 274:291 surface expression of the complex 274:306 Dysfunction in surface expression of the complex leads to Bernard-Soulier syndrome. 21908432 10 54 theme structural 1337:1346 arg1 proximity 1348:1356 The close structural proximity 1327:1356 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX 1327:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 9 55 theme GPIbβ 1266:1270 arg1 secretion/folding 1245:1261 normal secretion/folding 1238:1261 normal secretion/folding of GPIbβ(E) 1238:1273 Two mutations, A108P and P74R, were found to maintain normal secretion/folding of GPIbβ(E) but were unable to support GPIX surface expression. 21908432 9 56 theme normal 1238:1243 arg1 secretion/folding 1245:1261 normal secretion/folding 1238:1261 normal secretion/folding of GPIbβ(E) 1238:1273 Two mutations, A108P and P74R, were found to maintain normal secretion/folding of GPIbβ(E) but were unable to support GPIX surface expression. 21908432 0 57 theme Bernard-Soulier 61:75 arg1 syndrome 77:84 Bernard-Soulier syndrome 61:84 Bernard-Soulier syndrome 61:84 Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIbβ and a GPIbβ/GPIX chimera. 21908432 1 58 theme Platelet 148:155 arg1 complex 174:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex 148:180 Platelet GPIb-IX receptor complex has 3 subunits GPIbα, GPIbβ, and GPIX, which assemble with a ratio of 1:2:1. 21908432 10 59 theme E 1401:1401 arg1 interface 1404:1412 the GPIbβ(E) interface 1391:1412 the GPIbβ(E) interface with GPIX 1391:1422 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 21908432 10 60 theme GPIb-IX 1482:1488 arg1 complex 1490:1496 the GPIb-IX complex 1478:1496 the GPIb-IX complex 1478:1496 The close structural proximity of these mutations to Tyr106 and the GPIbβ(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex. 10684642 8 0 theme low 1105:1107 arg1 affinity 1118:1125 the low relative affinity 1101:1125 the low relative affinity for N-formylated tripeptides 1101:1154 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 9 1 theme overall 1473:1479 arg1 flexibility 1481:1491 the overall flexibility 1469:1491 the overall flexibility of the fold 1469:1503 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 4 2 theme lipocalin 592:600 arg1 fold 602:605 a typical lipocalin fold 582:605 a typical lipocalin fold 582:605 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 0 3 theme lipocalin 82:90 arg1 structure 36:44 the structure 32:44 the structure of neutrophil gelatinase associated lipocalin 32:90 Ligand preference inferred from the structure of neutrophil gelatinase associated lipocalin. 10684642 3 4 theme platelet 515:522 arg1 factor 535:540 platelet activating factor 515:540 platelet activating factor 515:540 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 4 5 theme typical 584:590 arg1 fold 602:605 a typical lipocalin fold 582:605 a typical lipocalin fold 582:605 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 6 6 theme sulfate 894:900 arg1 ions 902:905 sulfate ions 894:905 sulfate ions 894:905 The crystal structures also reveal either sulfate ions or an adventitiously copurified fatty acid bound in the binding site. 10684642 1 7 theme neutrophil 161:170 arg1 granules 172:179 neutrophil granules 161:179 neutrophil granules 161:179 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 8 8 theme relative 1109:1116 arg1 affinity 1118:1125 the low relative affinity 1101:1125 the low relative affinity for N-formylated tripeptides 1101:1154 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 9 theme protein 1282:1288 arg1 any 1208:1210 any 1208:1210 any 1208:1210 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 9 theme protein 1282:1288 arg1 ligand 1267:1272 the preferred ligand 1253:1272 the preferred ligand of this protein 1253:1288 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 9 theme protein 1282:1288 arg1 acid 1199:1202 the copurified fatty acid 1178:1202 the copurified fatty acid 1178:1202 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 9 theme protein 1282:1288 arg1 likely 1240:1245 likely 1240:1245 likely 1240:1245 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 9 10 theme crystal 1315:1321 arg1 structures 1323:1332 the crystal structures 1311:1332 the crystal structures 1311:1332 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 9 11 theme structure 1455:1463 arg1 details 1440:1446 the details 1436:1446 the details of the structure 1436:1463 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 9 11 theme structure 1455:1463 arg1 flexibility 1481:1491 the overall flexibility 1469:1491 the overall flexibility of the fold 1469:1503 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 8 12 theme N-formylated 1131:1142 arg1 tripeptides 1144:1154 N-formylated tripeptides 1131:1154 N-formylated tripeptides 1131:1154 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 9 13 theme details 1440:1446 arg1 terms 1422:1426 terms 1422:1426 terms of both the details of the structure and the overall flexibility of the fold 1422:1503 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 5 14 theme calyces 824:830 arg1 architecture 804:815 the overall architecture 792:815 the overall architecture of the calyces 792:830 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 5 14 theme calyces 824:830 arg1 different 841:849 different 841:849 different 841:849 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 4 15 theme large 633:637 arg1 site 668:671 an unusually large and atypically polar binding site 620:671 an unusually large and atypically polar binding site 620:671 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 9 16 theme fold 1500:1503 arg1 details 1440:1446 the details 1436:1446 the details of the structure 1436:1463 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 9 16 theme fold 1500:1503 arg1 flexibility 1481:1491 the overall flexibility 1469:1491 the overall flexibility of the fold 1469:1503 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 1 17 theme granules 172:179 arg1 constituent 146:156 a constituent 144:156 a constituent of neutrophil granules 144:179 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 17 theme granules 172:179 arg1 lipocalin 126:134 Neutrophil gelatinase associated lipocalin 93:134 Neutrophil gelatinase associated lipocalin (NGAL) 93:141 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 5 18 theme epididymal 724:733 arg1 protein 757:763 the epididymal retinoic acid-binding protein 720:763 the epididymal retinoic acid-binding protein 720:763 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 5 18 theme epididymal 724:733 arg1 lipocalin 774:782 another lipocalin 766:782 another lipocalin 766:782 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 3 19 theme inflammatory 375:386 arg1 responses 388:396 inflammatory responses 375:396 inflammatory responses 375:396 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 3 20 theme N-formylated 456:467 arg1 chemoattractants 425:440 neutrophil chemoattractants 414:440 neutrophil chemoattractants 414:440 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 3 20 theme N-formylated 456:467 arg1 tripeptides 469:479 N-formylated tripeptides 456:479 particularly N-formylated tripeptides 443:479 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 5 21 theme retinoic 735:742 arg1 protein 757:763 the epididymal retinoic acid-binding protein 720:763 the epididymal retinoic acid-binding protein 720:763 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 5 21 theme retinoic 735:742 arg1 lipocalin 774:782 another lipocalin 766:782 another lipocalin 766:782 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 1 22 theme Neutrophil 93:102 arg1 constituent 146:156 a constituent 144:156 a constituent of neutrophil granules 144:179 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 22 theme Neutrophil 93:102 arg1 NGAL 137:140 NGAL 137:140 NGAL 137:140 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 22 theme Neutrophil 93:102 arg1 member 187:192 a member 185:192 a member of the lipocalin family of binding proteins 185:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 22 theme Neutrophil 93:102 arg1 lipocalin 126:134 Neutrophil gelatinase associated lipocalin 93:134 Neutrophil gelatinase associated lipocalin (NGAL) 93:141 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 0 23 theme Ligand 0:5 arg1 preference 7:16 Ligand preference 0:16 Ligand preference 0:16 Ligand preference inferred from the structure of neutrophil gelatinase associated lipocalin. 10684642 4 24 theme NGAL 569:572 arg1 structures 555:564 The crystal structures 543:564 The crystal structures of NGAL 543:572 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 9 25 theme NGAL 1382:1385 arg1 structures 1323:1332 the crystal structures 1311:1332 the crystal structures 1311:1332 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 9 25 theme NGAL 1382:1385 arg1 structure 1369:1377 the recently reported solution structure 1338:1377 the recently reported solution structure of NGAL 1338:1385 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 1 26 theme gelatinase 104:113 arg1 constituent 146:156 a constituent 144:156 a constituent of neutrophil granules 144:179 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 26 theme gelatinase 104:113 arg1 NGAL 137:140 NGAL 137:140 NGAL 137:140 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 26 theme gelatinase 104:113 arg1 member 187:192 a member 185:192 a member of the lipocalin family of binding proteins 185:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 26 theme gelatinase 104:113 arg1 lipocalin 126:134 Neutrophil gelatinase associated lipocalin 93:134 Neutrophil gelatinase associated lipocalin (NGAL) 93:141 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 9 27 theme solution 1360:1367 arg1 structure 1369:1377 the recently reported solution structure 1338:1377 the recently reported solution structure of NGAL 1338:1385 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 5 28 theme overall 796:802 arg1 architecture 804:815 the overall architecture 792:815 the overall architecture of the calyces 792:830 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 5 28 theme overall 796:802 arg1 different 841:849 different 841:849 different 841:849 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 9 29 theme significant 1394:1404 arg1 differences 1406:1416 significant differences 1394:1416 significant differences 1394:1416 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 1 30 theme associated 115:124 arg1 constituent 146:156 a constituent 144:156 a constituent of neutrophil granules 144:179 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 30 theme associated 115:124 arg1 NGAL 137:140 NGAL 137:140 NGAL 137:140 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 30 theme associated 115:124 arg1 member 187:192 a member 185:192 a member of the lipocalin family of binding proteins 185:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 30 theme associated 115:124 arg1 lipocalin 126:134 Neutrophil gelatinase associated lipocalin 93:134 Neutrophil gelatinase associated lipocalin (NGAL) 93:141 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 8 31 theme proposed 1219:1226 arg1 ligands 1228:1234 the proposed ligands 1215:1234 the proposed ligands 1215:1234 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 5 32 theme NGAL 696:699 arg1 similar 709:715 similar 709:715 similar 709:715 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 5 32 theme NGAL 696:699 arg1 fold 688:691 The fold 684:691 The fold of NGAL 684:699 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 6 33 theme crystal 856:862 arg1 structures 864:873 The crystal structures 852:873 The crystal structures 852:873 The crystal structures also reveal either sulfate ions or an adventitiously copurified fatty acid bound in the binding site. 10684642 2 34 theme epithelial 274:283 arg1 cells 285:289 epithelial cells 274:289 epithelial cells in both inflammatory and neoplastic colorectal disease 274:344 NGAL can also be highly induced in epithelial cells in both inflammatory and neoplastic colorectal disease. 10684642 9 35 theme flexibility 1481:1491 arg1 terms 1422:1426 terms 1422:1426 terms of both the details of the structure and the overall flexibility of the fold 1422:1503 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 3 36 theme leukotriene 494:504 arg1 4 508:508 4 508:508 4 508:508 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 3 36 theme leukotriene 494:504 arg1 B 506:506 leukotriene B 494:506 leukotriene B(4) 494:509 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 4 37 theme polar 654:658 arg1 site 668:671 an unusually large and atypically polar binding site 620:671 an unusually large and atypically polar binding site 620:671 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 3 38 theme neutrophil 414:423 arg1 chemoattractants 425:440 neutrophil chemoattractants 414:440 neutrophil chemoattractants 414:440 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 3 38 theme neutrophil 414:423 arg1 tripeptides 469:479 N-formylated tripeptides 456:479 particularly N-formylated tripeptides 443:479 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 9 39 theme reported 1351:1358 arg1 structure 1369:1377 the recently reported solution structure 1338:1377 the recently reported solution structure of NGAL 1338:1385 Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. 10684642 1 40 theme lipocalin 201:209 arg1 family 211:216 the lipocalin family 197:216 the lipocalin family of binding proteins 197:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 0 41 theme neutrophil 49:58 arg1 lipocalin 82:90 neutrophil gelatinase associated lipocalin 49:90 neutrophil gelatinase associated lipocalin 49:90 Ligand preference inferred from the structure of neutrophil gelatinase associated lipocalin. 10684642 6 42 theme fatty 939:943 arg1 acid 945:948 an adventitiously copurified fatty acid 910:948 an adventitiously copurified fatty acid 910:948 The crystal structures also reveal either sulfate ions or an adventitiously copurified fatty acid bound in the binding site. 10684642 8 43 theme calyx 1083:1087 arg1 affinity 1118:1125 the low relative affinity 1101:1125 the low relative affinity for N-formylated tripeptides 1101:1154 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 43 theme calyx 1083:1087 arg1 shape 1050:1054 shape 1050:1054 shape 1050:1054 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 43 theme calyx 1083:1087 arg1 character 1061:1069 character 1061:1069 character 1061:1069 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 43 theme calyx 1083:1087 arg1 size 1044:1047 size 1044:1047 size 1044:1047 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 5 44 theme acid-binding 744:755 arg1 protein 757:763 the epididymal retinoic acid-binding protein 720:763 the epididymal retinoic acid-binding protein 720:763 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 5 44 theme acid-binding 744:755 arg1 lipocalin 774:782 another lipocalin 766:782 another lipocalin 766:782 The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. 10684642 7 45 theme added 1008:1012 arg1 tripeptides 1027:1037 added N-formylated tripeptides 1008:1037 added N-formylated tripeptides 1008:1037 Neither ligand is displaced by added N-formylated tripeptides. 10684642 2 46 theme colorectal 327:336 arg1 disease 338:344 both inflammatory and neoplastic colorectal disease 294:344 both inflammatory and neoplastic colorectal disease 294:344 NGAL can also be highly induced in epithelial cells in both inflammatory and neoplastic colorectal disease. 10684642 8 47 theme copurified 1182:1191 arg1 any 1208:1210 any 1208:1210 any 1208:1210 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 47 theme copurified 1182:1191 arg1 ligand 1267:1272 the preferred ligand 1253:1272 the preferred ligand of this protein 1253:1288 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 47 theme copurified 1182:1191 arg1 acid 1199:1202 the copurified fatty acid 1178:1202 the copurified fatty acid 1178:1202 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 47 theme copurified 1182:1191 arg1 likely 1240:1245 likely 1240:1245 likely 1240:1245 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 6 48 theme copurified 928:937 arg1 acid 945:948 an adventitiously copurified fatty acid 910:948 an adventitiously copurified fatty acid 910:948 The crystal structures also reveal either sulfate ions or an adventitiously copurified fatty acid bound in the binding site. 10684642 8 49 theme NGAL 1078:1081 arg1 calyx 1083:1087 the NGAL calyx 1074:1087 the NGAL calyx 1074:1087 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 2 50 theme neoplastic 316:325 arg1 disease 338:344 both inflammatory and neoplastic colorectal disease 294:344 both inflammatory and neoplastic colorectal disease 294:344 NGAL can also be highly induced in epithelial cells in both inflammatory and neoplastic colorectal disease. 10684642 4 51 theme crystal 547:553 arg1 structures 555:564 The crystal structures 543:564 The crystal structures of NGAL 543:572 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 7 52 theme N-formylated 1014:1025 arg1 tripeptides 1027:1037 added N-formylated tripeptides 1008:1037 added N-formylated tripeptides 1008:1037 Neither ligand is displaced by added N-formylated tripeptides. 10684642 1 53 theme family 211:216 arg1 member 187:192 a member 185:192 a member of the lipocalin family of binding proteins 185:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 1 53 theme family 211:216 arg1 lipocalin 126:134 Neutrophil gelatinase associated lipocalin 93:134 Neutrophil gelatinase associated lipocalin (NGAL) 93:141 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 2 54 theme inflammatory 299:310 arg1 disease 338:344 both inflammatory and neoplastic colorectal disease 294:344 both inflammatory and neoplastic colorectal disease 294:344 NGAL can also be highly induced in epithelial cells in both inflammatory and neoplastic colorectal disease. 10684642 8 55 theme fatty 1193:1197 arg1 any 1208:1210 any 1208:1210 any 1208:1210 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 55 theme fatty 1193:1197 arg1 ligand 1267:1272 the preferred ligand 1253:1272 the preferred ligand of this protein 1253:1288 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 55 theme fatty 1193:1197 arg1 acid 1199:1202 the copurified fatty acid 1178:1202 the copurified fatty acid 1178:1202 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 55 theme fatty 1193:1197 arg1 likely 1240:1245 likely 1240:1245 likely 1240:1245 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 0 56 theme associated 71:80 arg1 lipocalin 82:90 neutrophil gelatinase associated lipocalin 49:90 neutrophil gelatinase associated lipocalin 49:90 Ligand preference inferred from the structure of neutrophil gelatinase associated lipocalin. 10684642 3 57 theme activating 524:533 arg1 factor 535:540 platelet activating factor 515:540 platelet activating factor 515:540 NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. 10684642 4 58 theme binding 660:666 arg1 site 668:671 an unusually large and atypically polar binding site 620:671 an unusually large and atypically polar binding site 620:671 The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. 10684642 1 59 theme binding 221:227 arg1 proteins 229:236 binding proteins 221:236 binding proteins 221:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 10684642 0 60 theme gelatinase 60:69 arg1 lipocalin 82:90 neutrophil gelatinase associated lipocalin 49:90 neutrophil gelatinase associated lipocalin 49:90 Ligand preference inferred from the structure of neutrophil gelatinase associated lipocalin. 10684642 6 61 theme binding 963:969 arg1 site 971:974 the binding site 959:974 the binding site 959:974 The crystal structures also reveal either sulfate ions or an adventitiously copurified fatty acid bound in the binding site. 10684642 8 62 theme preferred 1257:1265 arg1 any 1208:1210 any 1208:1210 any 1208:1210 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 62 theme preferred 1257:1265 arg1 ligand 1267:1272 the preferred ligand 1253:1272 the preferred ligand of this protein 1253:1288 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 62 theme preferred 1257:1265 arg1 acid 1199:1202 the copurified fatty acid 1178:1202 the copurified fatty acid 1178:1202 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 8 62 theme preferred 1257:1265 arg1 likely 1240:1245 likely 1240:1245 likely 1240:1245 The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. 10684642 2 63 from cells 285:289 arg1 disease 338:344 both inflammatory and neoplastic colorectal disease 294:344 both inflammatory and neoplastic colorectal disease 294:344 NGAL can also be highly induced in epithelial cells in both inflammatory and neoplastic colorectal disease. 10684642 1 64 theme proteins 229:236 arg1 family 211:216 the lipocalin family 197:216 the lipocalin family of binding proteins 197:236 Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. 24190431 7 0 theme regulatory 1164:1173 arg1 cells 1177:1181 regulatory T cells 1164:1181 regulatory T cells relative to healthy individuals 1164:1213 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 6 1 theme * 1018:1018 arg1 blood 1001:1005 the peripheral blood 986:1005 the peripheral blood of HLA-DRB1*04:01 986:1023 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 2 2 theme citrullinated 309:321 arg1 aggrecan 323:330 citrullinated aggrecan 309:330 citrullinated aggrecan 309:330 We show how citrullinated aggrecan and vimentin epitopes bind to HLA-DRB1*04:01/04. 24190431 7 3 theme T 1175:1175 arg1 cells 1177:1181 regulatory T cells 1164:1181 regulatory T cells relative to healthy individuals 1164:1213 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 6 4 theme HLA-II 892:897 arg1 tetramers 899:907 HLA-II tetramers 892:907 HLA-II tetramers 892:907 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 8 5 from association 1264:1274 arg1 RA 1349:1350 RA 1349:1350 RA 1349:1350 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 7 6 theme healthy 1195:1201 arg1 individuals 1203:1213 healthy individuals 1195:1213 healthy individuals 1195:1213 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 4 7 from * 689:689 arg1 peptides 667:674 P4 arginine-containing peptides 644:674 P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04 644:726 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 6 8 theme T 975:975 arg1 cells 977:981 T cells 975:981 T cells in the peripheral blood of HLA-DRB1*04:01 975:1023 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 7 9 theme relative 1183:1190 arg1 cells 1177:1181 regulatory T cells 1164:1181 regulatory T cells relative to healthy individuals 1164:1213 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 1 10 theme shared 221:226 arg1 SE 252:253 SE 252:253 SE 252:253 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 10 theme shared 221:226 arg1 epitope 243:249 the shared susceptibility epitope 217:249 the shared susceptibility epitope (SE) 217:254 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 8 11 theme cell 1326:1329 arg1 autoreactivity 1331:1344 T cell autoreactivity 1324:1344 T cell autoreactivity 1324:1344 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 7 12 theme RA 1068:1069 arg1 patients 1071:1078 RA patients 1068:1078 RA patients 1068:1078 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 8 13 theme HLA-DRB1 1304:1311 arg1 locus 1313:1317 the HLA-DRB1 locus 1300:1317 the HLA-DRB1 locus 1300:1317 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 1 14 theme susceptibility 228:241 arg1 SE 252:253 SE 252:253 SE 252:253 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 14 theme susceptibility 228:241 arg1 epitope 243:249 the shared susceptibility epitope 217:249 the shared susceptibility epitope (SE) 217:254 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 5 15 theme T 844:844 arg1 cells 846:850 T cells 844:850 T cells 844:850 Citrullination altered protease susceptibility of vimentin, thereby generating self-epitopes that are presented to T cells in HLA-DRB1*04:01(+) individuals. 24190431 4 16 theme Peptide 611:617 arg1 studies 627:633 Peptide elution studies 611:633 Peptide elution studies 611:633 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 3 17 theme P4 496:497 arg1 pocket 499:504 the electronegative P4 pocket 476:504 the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph 476:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 4 18 theme arginine-containing 647:665 arg1 peptides 667:674 P4 arginine-containing peptides 644:674 P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04 644:726 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 0 19 theme rheumatoid 81:90 arg1 arthritis 92:100 rheumatoid arthritis 81:100 rheumatoid arthritis 81:100 A molecular basis for the association of the HLA-DRB1 locus, citrullination, and rheumatoid arthritis. 24190431 7 20 theme cell 1096:1099 arg1 numbers 1101:1107 autoreactive T cell numbers 1081:1107 autoreactive T cell numbers 1081:1107 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 4 21 theme elution 619:625 arg1 studies 627:633 Peptide elution studies 611:633 Peptide elution studies 611:633 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 6 22 theme peripheral 990:999 arg1 blood 1001:1005 the peripheral blood 986:1005 the peripheral blood of HLA-DRB1*04:01 986:1023 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 4 23 theme P4 644:645 arg1 peptides 667:674 P4 arginine-containing peptides 644:674 P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04 644:726 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 0 24 theme molecular 2:10 arg1 basis 12:16 A molecular basis 0:16 A molecular basis for the association of the HLA-DRB1 locus 0:58 A molecular basis for the association of the HLA-DRB1 locus, citrullination, and rheumatoid arthritis. 24190431 6 25 from cells 977:981 arg1 blood 1001:1005 the peripheral blood 986:1005 the peripheral blood of HLA-DRB1*04:01 986:1023 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 3 26 theme * 534:534 arg1 allomorph 541:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 7 27 theme disease 1125:1131 arg1 activity 1133:1140 disease activity 1125:1140 disease activity 1125:1140 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 8 28 from RA 1349:1350 arg1 understanding 1243:1255 our understanding 1239:1255 our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA 1239:1350 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 3 29 theme 04:02 535:539 arg1 allomorph 541:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 2 30 theme vimentin 336:343 arg1 epitopes 345:352 vimentin epitopes 336:352 vimentin epitopes 336:352 We show how citrullinated aggrecan and vimentin epitopes bind to HLA-DRB1*04:01/04. 24190431 8 31 theme association 1264:1274 arg1 understanding 1243:1255 our understanding 1239:1255 our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA 1239:1350 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 4 32 from * 718:718 arg1 peptides 667:674 P4 arginine-containing peptides 644:674 P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04 644:726 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 3 33 theme RA-resistant 513:524 arg1 allomorph 541:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 1 34 theme leukocyte 167:175 arg1 antigen 177:183 human leukocyte antigen 161:183 the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens 157:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 34 theme leukocyte 167:175 arg1 HLA 186:188 HLA 186:188 HLA 186:188 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 8 35 theme T 1324:1324 arg1 autoreactivity 1331:1344 T cell autoreactivity 1324:1344 T cell autoreactivity 1324:1344 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 6 36 theme aggrecan-specific 950:966 arg1 individuals 1052:1062 citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals 922:1062 citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals 922:1062 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 3 37 theme HLA-DRB1 526:533 arg1 allomorph 541:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 the RA-resistant HLA-DRB1*04:02 allomorph 509:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 1 38 theme antigen 177:183 arg1 locus 196:200 the human leukocyte antigen (HLA)-DRB1 locus 157:200 the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens 157:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 5 39 theme vimentin 779:786 arg1 susceptibility 761:774 protease susceptibility 752:774 protease susceptibility of vimentin 752:786 Citrullination altered protease susceptibility of vimentin, thereby generating self-epitopes that are presented to T cells in HLA-DRB1*04:01(+) individuals. 24190431 1 40 theme human 161:165 arg1 antigen 177:183 human leukocyte antigen 161:183 the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens 157:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 40 theme human 161:165 arg1 HLA 186:188 HLA 186:188 HLA 186:188 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 6 41 theme HLA-DRB1 1010:1017 arg1 * 1018:1018 HLA-DRB1*04:01 1010:1023 HLA-DRB1*04:01 1010:1023 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 5 42 theme HLA-DRB1 855:862 arg1 * 863:863 HLA-DRB1*04:01 855:868 HLA-DRB1*04:01 855:868 Citrullination altered protease susceptibility of vimentin, thereby generating self-epitopes that are presented to T cells in HLA-DRB1*04:01(+) individuals. 24190431 6 43 theme vimentin- 936:944 arg1 individuals 1052:1062 citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals 922:1062 citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals 922:1062 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 8 44 from understanding 1243:1255 arg1 RA 1349:1350 RA 1349:1350 RA 1349:1350 These findings reshape our understanding of the association between citrullination, the HLA-DRB1 locus, and T cell autoreactivity in RA. 24190431 7 45 from deficient 1151:1159 arg1 cells 1177:1181 regulatory T cells 1164:1181 regulatory T cells relative to healthy individuals 1164:1213 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 2 46 theme HLA-DRB1 362:369 arg1 * 370:370 HLA-DRB1*04:01/04 362:378 HLA-DRB1*04:01/04 362:378 We show how citrullinated aggrecan and vimentin epitopes bind to HLA-DRB1*04:01/04. 24190431 7 47 theme autoreactive 1081:1092 arg1 numbers 1101:1107 autoreactive T cell numbers 1081:1107 autoreactive T cell numbers 1081:1107 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 6 48 theme citrullinated 922:934 arg1 individuals 1052:1062 citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals 922:1062 citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals 922:1062 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 5 49 theme protease 752:759 arg1 susceptibility 761:774 protease susceptibility 752:774 protease susceptibility of vimentin 752:786 Citrullination altered protease susceptibility of vimentin, thereby generating self-epitopes that are presented to T cells in HLA-DRB1*04:01(+) individuals. 24190431 3 50 theme allomorph 541:549 arg1 pocket 499:504 the electronegative P4 pocket 476:504 the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph 476:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 3 51 theme * 457:457 arg1 pocket 439:444 the electropositive P4 pocket 416:444 the electropositive P4 pocket of HLA-DRB1*04:01/04 416:465 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 7 52 theme T 1094:1094 arg1 numbers 1101:1107 autoreactive T cell numbers 1081:1107 autoreactive T cell numbers 1081:1107 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 6 53 dep aggrecan-specific 950:966 arg1 cells 977:981 T cells 975:981 T cells in the peripheral blood of HLA-DRB1*04:01 975:1023 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 6 53 dep aggrecan-specific 950:966 arg1 CD4 968:970 CD4 968:970 CD4 968:970 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 6 53 dep aggrecan-specific 950:966 arg1 healthy 1044:1050 healthy 1044:1050 healthy 1044:1050 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 6 53 dep aggrecan-specific 950:966 arg1 RA-affected 1028:1038 RA-affected 1028:1038 RA-affected 1028:1038 Using HLA-II tetramers, we observed citrullinated vimentin- and aggrecan-specific CD4(+) T cells in the peripheral blood of HLA-DRB1*04:01(+) RA-affected and healthy individuals. 24190431 1 54 theme Rheumatoid 103:112 arg1 arthritis 114:122 Rheumatoid arthritis 103:122 Rheumatoid arthritis (RA) 103:127 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 54 theme Rheumatoid 103:112 arg1 RA 125:126 RA 125:126 RA 125:126 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 3 55 theme citrulline-containing 579:599 arg1 epitopes 601:608 citrulline-containing epitopes 579:608 citrulline-containing epitopes 579:608 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 4 56 theme HLA-DRB1 681:688 arg1 * 689:689 HLA-DRB1*04:02 681:694 HLA-DRB1*04:02 681:694 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 4 57 theme HLA-DRB1 710:717 arg1 * 718:718 HLA-DRB1*04:01/04 710:726 HLA-DRB1*04:01/04 710:726 Peptide elution studies revealed P4 arginine-containing peptides from HLA-DRB1*04:02, but not from HLA-DRB1*04:01/04. 24190431 7 58 from cells 1177:1181 arg1 deficient 1151:1159 deficient 1151:1159 deficient 1151:1159 In RA patients, autoreactive T cell numbers correlated with disease activity and were deficient in regulatory T cells relative to healthy individuals. 24190431 3 59 theme HLA-DRB1 449:456 arg1 * 457:457 HLA-DRB1*04:01/04 449:465 HLA-DRB1*04:01/04 449:465 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 1 60 theme -DRB1 190:194 arg1 locus 196:200 the human leukocyte antigen (HLA)-DRB1 locus 157:200 the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens 157:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 0 61 theme locus 54:58 arg1 association 26:36 the association 22:36 the association of the HLA-DRB1 locus 22:58 A molecular basis for the association of the HLA-DRB1 locus, citrullination, and rheumatoid arthritis. 24190431 3 62 theme electropositive 420:434 arg1 pocket 439:444 the electropositive P4 pocket 416:444 the electropositive P4 pocket of HLA-DRB1*04:01/04 416:465 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 3 63 theme P4 436:437 arg1 pocket 439:444 the electropositive P4 pocket 416:444 the electropositive P4 pocket of HLA-DRB1*04:01/04 416:465 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 24190431 5 64 theme * 863:863 arg1 individuals 873:883 HLA-DRB1*04:01(+) individuals 855:883 HLA-DRB1*04:01(+) individuals 855:883 Citrullination altered protease susceptibility of vimentin, thereby generating self-epitopes that are presented to T cells in HLA-DRB1*04:01(+) individuals. 24190431 0 65 theme HLA-DRB1 45:52 arg1 locus 54:58 the HLA-DRB1 locus 41:58 the HLA-DRB1 locus 41:58 A molecular basis for the association of the HLA-DRB1 locus, citrullination, and rheumatoid arthritis. 24190431 1 66 theme self-antigens 282:294 arg1 SE 252:253 SE 252:253 SE 252:253 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 66 theme self-antigens 282:294 arg1 epitope 243:249 the shared susceptibility epitope 217:249 the shared susceptibility epitope (SE) 217:254 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 66 theme self-antigens 282:294 arg1 citrullination 264:277 the citrullination 260:277 the citrullination of self-antigens 260:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 67 contain possesses 207:215 arg1 locus 196:200 the human leukocyte antigen (HLA)-DRB1 locus 157:200 the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens 157:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 67 contain possesses 207:215 arg2 epitope 243:249 the shared susceptibility epitope 217:249 the shared susceptibility epitope (SE) 217:254 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 67 contain possesses 207:215 arg2 citrullination 264:277 the citrullination 260:277 the citrullination of self-antigens 260:294 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 1 67 contain possesses 207:215 arg2 SE 252:253 SE 252:253 SE 252:253 Rheumatoid arthritis (RA) is strongly associated with the human leukocyte antigen (HLA)-DRB1 locus that possesses the shared susceptibility epitope (SE) and the citrullination of self-antigens. 24190431 3 68 theme electronegative 480:494 arg1 pocket 499:504 the electronegative P4 pocket 476:504 the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph 476:549 Citrulline was accommodated within the electropositive P4 pocket of HLA-DRB1*04:01/04, whereas the electronegative P4 pocket of the RA-resistant HLA-DRB1*04:02 allomorph interacted with arginine or citrulline-containing epitopes. 19303388 3 0 theme MS 626:627 arg1 autoantigen 629:639 a candidate MS autoantigen 614:639 a candidate MS autoantigen 614:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 0 theme MS 626:627 arg1 protein 605:611 myelin basic protein 592:611 myelin basic protein 592:611 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 1 theme bacteria 455:462 arg1 classes 444:450 several major classes 430:450 several major classes of bacteria 430:462 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 2 from disease 484:490 arg1 mice 505:508 humanized mice 495:508 humanized mice 495:508 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 0 3 theme microbial 81:89 arg1 peptides 91:98 common microbial peptides 74:98 common microbial peptides 74:98 T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. 19303388 5 4 theme Biophysical 828:838 arg1 studies 840:846 Biophysical studies 828:846 Biophysical studies 828:846 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 6 5 theme possible 1015:1022 arg1 explanation 1024:1034 a possible explanation 1013:1034 a possible explanation for the difficulty in incriminating individual infections in the development of MS 1013:1117 Thus, these data suggest a possible explanation for the difficulty in incriminating individual infections in the development of MS. 19303388 4 6 theme TCR 811:813 arg1 recognition 815:825 degenerate TCR recognition 800:825 degenerate TCR recognition 800:825 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 2 7 theme them 277:280 arg1 none 269:272 none 269:272 none of them 269:280 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 4 8 theme degenerate 800:809 arg1 recognition 815:825 degenerate TCR recognition 800:825 degenerate TCR recognition 800:825 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 3 9 theme humanized 495:503 arg1 mice 505:508 humanized mice 495:508 humanized mice 495:508 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 4 10 theme Structural 642:651 arg1 analysis 653:660 Structural analysis 642:660 Structural analysis 642:660 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 3 11 theme MS-like 476:482 arg1 disease 484:490 MS-like disease 476:490 MS-like disease in humanized mice 476:508 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 1 12 theme multiple 165:172 arg1 MS 185:186 MS 185:186 MS 185:186 Environmental factors account for 75% of the risk of developing multiple sclerosis (MS). 19303388 1 12 theme multiple 165:172 arg1 sclerosis 174:182 multiple sclerosis 165:182 multiple sclerosis (MS) 165:187 Environmental factors account for 75% of the risk of developing multiple sclerosis (MS). 19303388 4 13 theme microbial 765:773 arg1 antigens 775:782 microbial antigens 765:782 microbial antigens 765:782 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 4 14 theme structural 706:715 arg1 mimicry 717:723 structural mimicry 706:723 structural mimicry of a binding hotspot shared by self and microbial antigens 706:782 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 3 15 theme microbial 401:409 arg1 peptide 411:417 a microbial peptide 399:417 a microbial peptide 399:417 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 0 16 theme cell-mediated 2:14 arg1 disease 27:33 T cell-mediated autoimmune disease 0:33 T cell-mediated autoimmune disease 0:33 T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. 19303388 3 17 from protein 605:611 arg1 peptide 579:585 a peptide 577:585 a peptide from myelin basic protein, a candidate MS autoantigen 577:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 5 18 theme mimicry 935:941 arg1 peptide 944:950 the microbial (mimicry) peptide 920:950 the microbial (mimicry) peptide 920:950 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 5 19 theme TCR 877:879 arg1 affinity 889:896 the autoreactive TCR binding affinity 860:896 the autoreactive TCR binding affinity 860:896 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 5 19 theme TCR 877:879 arg1 lower 910:914 lower 910:914 lower 910:914 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 1 20 theme Environmental 101:113 arg1 factors 115:121 Environmental factors 101:121 Environmental factors 101:121 Environmental factors account for 75% of the risk of developing multiple sclerosis (MS). 19303388 2 21 theme different 348:356 arg1 infections 358:367 so many different infections 340:367 so many different infections 340:367 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 4 22 theme hotspot 738:744 arg1 mimicry 717:723 structural mimicry 706:723 structural mimicry of a binding hotspot shared by self and microbial antigens 706:782 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 5 23 theme autoreactive 864:875 arg1 affinity 889:896 the autoreactive TCR binding affinity 860:896 the autoreactive TCR binding affinity 860:896 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 5 23 theme autoreactive 864:875 arg1 lower 910:914 lower 910:914 lower 910:914 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 5 24 theme autoantigenic 965:977 arg1 peptide 979:985 the autoantigenic peptide 961:985 the autoantigenic peptide 961:985 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 3 25 theme common 420:425 arg1 peptide 411:417 a microbial peptide 399:417 a microbial peptide 399:417 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 0 26 theme autoimmune 16:25 arg1 disease 27:33 T cell-mediated autoimmune disease 0:33 T cell-mediated autoimmune disease 0:33 T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. 19303388 5 27 theme binding 881:887 arg1 affinity 889:896 the autoreactive TCR binding affinity 860:896 the autoreactive TCR binding affinity 860:896 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 5 27 theme binding 881:887 arg1 lower 910:914 lower 910:914 lower 910:914 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 6 28 theme individual 1072:1081 arg1 infections 1083:1092 individual infections 1072:1092 individual infections 1072:1092 Thus, these data suggest a possible explanation for the difficulty in incriminating individual infections in the development of MS. 19303388 3 29 theme cell 536:539 arg1 TCR 551:553 TCR 551:553 TCR 551:553 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 29 theme cell 536:539 arg1 receptor 541:548 a T cell receptor 532:548 a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen 532:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 0 30 theme low-affinity 42:53 arg1 crossreactivity 55:69 low-affinity crossreactivity 42:69 low-affinity crossreactivity to common microbial peptides 42:98 T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. 19303388 6 31 theme MS 1116:1117 arg1 development 1101:1111 the development 1097:1111 the development of MS 1097:1117 Thus, these data suggest a possible explanation for the difficulty in incriminating individual infections in the development of MS. 19303388 3 32 theme basic 599:603 arg1 autoantigen 629:639 a candidate MS autoantigen 614:639 a candidate MS autoantigen 614:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 32 theme basic 599:603 arg1 protein 605:611 myelin basic protein 592:611 myelin basic protein 592:611 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 33 theme T 534:534 arg1 TCR 551:553 TCR 551:553 TCR 551:553 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 33 theme T 534:534 arg1 receptor 541:548 a T cell receptor 532:548 a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen 532:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 4 34 theme binding 730:736 arg1 hotspot 738:744 a binding hotspot 728:744 a binding hotspot shared by self and microbial antigens 728:782 Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. 19303388 2 35 theme Numerous 190:197 arg1 triggers 255:262 environmental disease triggers 233:262 environmental disease triggers 233:262 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 2 35 theme Numerous 190:197 arg1 infections 199:208 Numerous infections 190:208 Numerous infections 190:208 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 0 36 dep disease 27:33 arg1 crossreactivity 55:69 low-affinity crossreactivity 42:69 low-affinity crossreactivity to common microbial peptides 42:98 T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. 19303388 3 37 theme several 430:436 arg1 classes 444:450 several major classes 430:450 several major classes of bacteria 430:462 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 38 theme major 438:442 arg1 classes 444:450 several major classes 430:450 several major classes of bacteria 430:462 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 0 39 theme common 74:79 arg1 peptides 91:98 common microbial peptides 74:98 common microbial peptides 74:98 T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. 19303388 3 40 theme myelin 592:597 arg1 autoantigen 629:639 a candidate MS autoantigen 614:639 a candidate MS autoantigen 614:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 40 theme myelin 592:597 arg1 protein 605:611 myelin basic protein 592:611 myelin basic protein 592:611 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 2 41 theme disease 247:253 arg1 triggers 255:262 environmental disease triggers 233:262 environmental disease triggers 233:262 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 2 41 theme disease 247:253 arg1 infections 199:208 Numerous infections 190:208 Numerous infections 190:208 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 2 42 theme many 343:346 arg1 infections 358:367 so many different infections 340:367 so many different infections 340:367 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 5 43 theme microbial 924:932 arg1 peptide 944:950 the microbial (mimicry) peptide 920:950 the microbial (mimicry) peptide 920:950 Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. 19303388 3 44 theme candidate 616:624 arg1 autoantigen 629:639 a candidate MS autoantigen 614:639 a candidate MS autoantigen 614:639 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 3 44 theme candidate 616:624 arg1 protein 605:611 myelin basic protein 592:611 myelin basic protein 592:611 We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. 19303388 2 45 theme environmental 233:245 arg1 triggers 255:262 environmental disease triggers 233:262 environmental disease triggers 233:262 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 2 45 theme environmental 233:245 arg1 infections 199:208 Numerous infections 190:208 Numerous infections 190:208 Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. 19303388 1 46 theme risk 146:149 arg1 % 137:137 75% 135:137 75% of the risk of developing multiple sclerosis (MS) 135:187 Environmental factors account for 75% of the risk of developing multiple sclerosis (MS). 19303388 1 46 theme risk 146:149 arg1 risk 146:149 the risk 142:149 the risk of developing multiple sclerosis (MS) 142:187 Environmental factors account for 75% of the risk of developing multiple sclerosis (MS). 9501082 8 0 theme alpha6 1107:1112 arg1 lactose 1139:1145 lactose 1139:1145 lactose 1139:1145 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 8 0 theme alpha6 1107:1112 arg1 subunits 1114:1121 alpha2 and alpha6 subunits 1096:1121 alpha2 and alpha6 subunits 1096:1121 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 8 0 theme alpha6 1107:1112 arg1 peptide 1128:1134 RGD peptide 1124:1134 RGD peptide 1124:1134 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 1 1 theme 92 309:310 arg1 kDa 312:314 kDa 312:314 kDa 312:314 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 7 2 with strength 990:997 arg1 laminin 1004:1010 laminin 1004:1010 laminin 1004:1010 The protein also mediated adhesion of cell lines at comparable strength with laminin. 9501082 10 3 located detected 1362:1369 arg2 secretion 1309:1317 variable secretion 1300:1317 variable secretion 1300:1317 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 10 3 located detected 1362:1369 arg1 matrix 1392:1397 the extracellular matrix 1374:1397 the extracellular matrix of several mouse tissues 1374:1422 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 0 4 theme matrix 70:75 arg1 protein 14:20 Mac-2 binding protein 0:20 Mac-2 binding protein 0:20 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 0 4 theme matrix 70:75 arg1 protein 41:47 a cell-adhesive protein 25:47 a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin 25:175 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 1 5 theme kDa 312:314 arg1 subunit 316:322 a 92 kDa subunit 307:322 a 92 kDa subunit 307:322 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 10 6 theme tissues 1416:1422 arg1 matrix 1392:1397 the extracellular matrix 1374:1397 the extracellular matrix of several mouse tissues 1374:1422 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 8 7 theme alpha2 1096:1101 arg1 lactose 1139:1145 lactose 1139:1145 lactose 1139:1145 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 8 7 theme alpha2 1096:1101 arg1 subunits 1114:1121 alpha2 and alpha6 subunits 1096:1121 alpha2 and alpha6 subunits 1096:1121 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 8 7 theme alpha2 1096:1101 arg1 peptide 1128:1134 RGD peptide 1124:1134 RGD peptide 1124:1134 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 9 8 theme cell 1167:1170 arg1 adhesion 1172:1179 cell adhesion 1167:1179 cell adhesion of M2BP 1167:1187 This distinguishes cell adhesion of M2BP from that of laminin and excludes involvement of lactose-binding galectin-3. 9501082 10 9 theme mouse 1410:1414 arg1 tissues 1416:1422 several mouse tissues 1402:1422 several mouse tissues 1402:1422 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 4 10 theme molar 661:665 arg1 masses 667:672 molar masses 661:672 molar masses of 1000-1500 kDa 661:689 Ultracentrifugation showed non-covalent association into oligomers with molar masses of 1000-1500 kDa. 9501082 2 11 theme immunological 485:497 arg1 analysis 499:506 immunological analysis 485:506 immunological analysis 485:506 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 10 12 theme several 1402:1408 arg1 tissues 1416:1422 several mouse tissues 1402:1422 several mouse tissues 1402:1422 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 5 13 theme nm 760:761 arg1 diameters 741:749 diameters 741:749 diameters of 30-40 nm 741:761 Electron microscopy showed ring-like shapes with diameters of 30-40 nm. 9501082 1 14 theme culture 255:261 arg1 medium 263:268 the culture medium 251:268 the culture medium of 293 kidney cells 251:288 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 6 15 theme fibrillar 860:868 arg1 collagens 870:878 fibrillar collagens I and III 860:888 fibrillar collagens I and III 860:888 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 15 theme fibrillar 860:868 arg1 III 886:888 III 886:888 III 886:888 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 15 theme fibrillar 860:868 arg1 I 880:880 I 880:880 I 880:880 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 5 16 with shapes 729:734 arg1 diameters 741:749 diameters 741:749 diameters of 30-40 nm 741:761 Electron microscopy showed ring-like shapes with diameters of 30-40 nm. 9501082 2 17 theme native 355:360 arg1 state 362:366 a native state 353:366 a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure 353:451 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 0 18 theme ring-like 103:111 arg1 structures 113:122 ring-like structures 103:122 ring-like structures 103:122 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 1 19 theme Human 178:182 arg1 M2BP 207:210 M2BP 207:210 M2BP 207:210 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 1 19 theme Human 178:182 arg1 protein 198:204 Human Mac-2 binding protein 178:204 Human Mac-2 binding protein (M2BP) 178:211 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 0 20 theme binding 6:12 arg1 protein 14:20 Mac-2 binding protein 0:20 Mac-2 binding protein 0:20 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 0 20 theme binding 6:12 arg1 protein 41:47 a cell-adhesive protein 25:47 a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin 25:175 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 1 21 from medium 263:268 arg1 form 241:244 recombinant form 229:244 recombinant form from the culture medium of 293 kidney cells 229:288 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 0 22 theme extracellular 56:68 arg1 matrix 70:75 the extracellular matrix 52:75 the extracellular matrix 52:75 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 1 23 theme Mac-2 184:188 arg1 M2BP 207:210 M2BP 207:210 M2BP 207:210 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 1 23 theme Mac-2 184:188 arg1 protein 198:204 Human Mac-2 binding protein 178:204 Human Mac-2 binding protein (M2BP) 178:211 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 0 24 theme Mac-2 0:4 arg1 protein 14:20 Mac-2 binding protein 0:20 Mac-2 binding protein 0:20 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 0 24 theme Mac-2 0:4 arg1 protein 41:47 a cell-adhesive protein 25:47 a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin 25:175 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 6 25 theme solid-phase 778:788 arg1 assays 790:795 solid-phase assays 778:795 solid-phase assays 778:795 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 10 26 theme variable 1300:1307 arg1 secretion 1309:1317 variable secretion 1300:1317 variable secretion 1300:1317 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 1 27 theme binding 190:196 arg1 M2BP 207:210 M2BP 207:210 M2BP 207:210 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 1 27 theme binding 190:196 arg1 protein 198:204 Human Mac-2 binding protein 178:204 Human Mac-2 binding protein (M2BP) 178:211 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 10 28 theme extracellular 1378:1390 arg1 matrix 1392:1397 the extracellular matrix 1374:1397 the extracellular matrix of several mouse tissues 1374:1422 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 6 29 dep collagens 800:808 arg1 IV 810:811 IV 810:811 IV 810:811 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 29 dep collagens 800:808 arg1 fibronectin 824:834 fibronectin 824:834 fibronectin 824:834 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 29 dep collagens 800:808 arg1 collagens 800:808 collagens IV, V and VI, fibronectin and nidogen 800:846 collagens IV, V and VI, fibronectin and nidogen 800:846 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 29 dep collagens 800:808 arg1 V 814:814 V 814:814 V 814:814 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 2 30 theme CD 384:385 arg1 spectroscopy 387:398 CD spectroscopy 384:398 CD spectroscopy 384:398 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 6 31 dep fibronectin 824:834 arg1 VI 820:821 VI 820:821 VI 820:821 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 9 32 theme M2BP 1184:1187 arg1 adhesion 1172:1179 cell adhesion 1167:1179 cell adhesion of M2BP 1167:1187 This distinguishes cell adhesion of M2BP from that of laminin and excludes involvement of lactose-binding galectin-3. 9501082 4 33 theme non-covalent 616:627 arg1 association 629:639 non-covalent association 616:639 non-covalent association into oligomers with molar masses of 1000-1500 kDa 616:689 Ultracentrifugation showed non-covalent association into oligomers with molar masses of 1000-1500 kDa. 9501082 0 34 theme cell-adhesive 27:39 arg1 protein 14:20 Mac-2 binding protein 0:20 Mac-2 binding protein 0:20 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 0 34 theme cell-adhesive 27:39 arg1 protein 41:47 a cell-adhesive protein 25:47 a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin 25:175 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 1 35 theme kidney 277:282 arg1 cells 284:288 293 kidney cells 273:288 293 kidney cells 273:288 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 5 36 theme ring-like 719:727 arg1 shapes 729:734 ring-like shapes 719:734 ring-like shapes with diameters of 30-40 nm 719:761 Electron microscopy showed ring-like shapes with diameters of 30-40 nm. 9501082 9 37 theme lactose-binding 1238:1252 arg1 galectin-3 1254:1263 lactose-binding galectin-3 1238:1263 lactose-binding galectin-3 1238:1263 This distinguishes cell adhesion of M2BP from that of laminin and excludes involvement of lactose-binding galectin-3. 9501082 10 38 theme Immunological 1266:1278 arg1 assays 1280:1285 Immunological assays 1266:1285 Immunological assays 1266:1285 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 2 39 theme beta-type 433:441 arg1 structure 443:451 alpha-helical and beta-type structure 415:451 alpha-helical and beta-type structure 415:451 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 1 40 theme cells 284:288 arg1 medium 263:268 the culture medium 251:268 the culture medium of 293 kidney cells 251:288 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 9501082 0 41 theme beta1 134:138 arg1 integrins 140:148 beta1 integrins 134:148 beta1 integrins 134:148 Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin. 9501082 2 42 dep state 362:366 arg1 indicated 371:379 indicated 371:379 indicated 371:379 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 7 43 theme comparable 979:988 arg1 strength 990:997 comparable strength 979:997 comparable strength with laminin 979:1010 The protein also mediated adhesion of cell lines at comparable strength with laminin. 9501082 8 44 theme RGD 1124:1126 arg1 subunits 1114:1121 alpha2 and alpha6 subunits 1096:1121 alpha2 and alpha6 subunits 1096:1121 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 8 44 theme RGD 1124:1126 arg1 peptide 1128:1134 RGD peptide 1124:1134 RGD peptide 1124:1134 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 6 45 theme membrane 908:915 arg1 proteins 917:924 other basement membrane proteins 893:924 other basement membrane proteins 893:924 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 10 46 theme cultured 1322:1329 arg1 cells 1337:1341 cultured human cells 1322:1341 cultured human cells of M2BP 1322:1349 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 2 47 theme alpha-helical 415:427 arg1 structure 443:451 alpha-helical and beta-type structure 415:451 alpha-helical and beta-type structure 415:451 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 6 48 theme basement 899:906 arg1 proteins 917:924 other basement membrane proteins 893:924 other basement membrane proteins 893:924 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 8 49 theme beta1 1070:1074 arg1 subunits 1076:1083 integrin beta1 subunits 1061:1083 integrin beta1 subunits 1061:1083 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 7 50 theme lines 970:974 arg1 adhesion 953:960 adhesion 953:960 adhesion of cell lines 953:974 The protein also mediated adhesion of cell lines at comparable strength with laminin. 9501082 6 51 theme other 893:897 arg1 proteins 917:924 other basement membrane proteins 893:924 other basement membrane proteins 893:924 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 8 52 theme integrin 1061:1068 arg1 subunits 1076:1083 integrin beta1 subunits 1061:1083 integrin beta1 subunits 1061:1083 Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. 9501082 3 53 mod modified 523:530 arg1 It 509:510 It 509:510 It 509:510 It was highly modified by N- and O-glycosylation but not by glycosaminoglycans. 9501082 3 53 mod modified 523:530 arg3 N- 535:536 N- 535:536 N- 535:536 It was highly modified by N- and O-glycosylation but not by glycosaminoglycans. 9501082 5 54 theme Electron 692:699 arg1 microscopy 701:710 Electron microscopy 692:710 Electron microscopy 692:710 Electron microscopy showed ring-like shapes with diameters of 30-40 nm. 9501082 4 55 with association 629:639 arg1 masses 667:672 molar masses 661:672 molar masses of 1000-1500 kDa 661:689 Ultracentrifugation showed non-covalent association into oligomers with molar masses of 1000-1500 kDa. 9501082 10 56 theme human 1331:1335 arg1 cells 1337:1341 cultured human cells 1322:1341 cultured human cells of M2BP 1322:1349 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 2 57 theme protease 461:468 arg1 resistance 470:479 protease resistance 461:479 protease resistance 461:479 The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. 9501082 10 58 theme M2BP 1346:1349 arg1 cells 1337:1341 cultured human cells 1322:1341 cultured human cells of M2BP 1322:1349 Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues. 9501082 7 59 theme cell 965:968 arg1 lines 970:974 cell lines 965:974 cell lines 965:974 The protein also mediated adhesion of cell lines at comparable strength with laminin. 9501082 4 60 theme kDa 687:689 arg1 masses 667:672 molar masses 661:672 molar masses of 1000-1500 kDa 661:689 Ultracentrifugation showed non-covalent association into oligomers with molar masses of 1000-1500 kDa. 9501082 6 61 dep collagens 870:878 arg1 collagens 870:878 fibrillar collagens I and III 860:888 fibrillar collagens I and III 860:888 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 61 dep collagens 870:878 arg1 III 886:888 III 886:888 III 886:888 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 6 61 dep collagens 870:878 arg1 I 880:880 I 880:880 I 880:880 M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. 9501082 9 62 theme galectin-3 1254:1263 arg1 involvement 1223:1233 involvement 1223:1233 involvement of lactose-binding galectin-3 1223:1263 This distinguishes cell adhesion of M2BP from that of laminin and excludes involvement of lactose-binding galectin-3. 9501082 1 63 theme recombinant 229:239 arg1 form 241:244 recombinant form 229:244 recombinant form from the culture medium of 293 kidney cells 229:288 Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. 12370246 1 0 theme epithelial 249:258 arg1 Golgi 236:240 the Golgi 232:240 the Golgi (e.g., epithelial MDCK cells) 232:270 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 1 0 theme epithelial 249:258 arg1 cells 265:269 epithelial MDCK cells 249:269 epithelial MDCK cells 249:269 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 0 1 from cells 124:128 arg1 MAL2 0:3 MAL2 0:3 MAL2 0:3 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 1 from cells 124:128 arg1 component 62:70 an essential component 49:70 an essential component of the machinery for transcytosis in hepatoma HepG2 cells 49:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 1 2 theme MDCK 260:263 arg1 Golgi 236:240 the Golgi 232:240 the Golgi (e.g., epithelial MDCK cells) 232:270 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 1 2 theme MDCK 260:263 arg1 cells 265:269 epithelial MDCK cells 249:269 epithelial MDCK cells 249:269 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 2 3 from route 427:431 arg1 cells 441:445 MDCK cells 436:445 MDCK cells 436:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 3 4 theme MAL 524:526 arg1 family 536:541 the MAL protein family 520:541 the MAL protein family 520:541 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 0 5 theme essential 52:60 arg1 MAL2 0:3 MAL2 0:3 MAL2 0:3 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 5 theme essential 52:60 arg1 component 62:70 an essential component 49:70 an essential component of the machinery for transcytosis in hepatoma HepG2 cells 49:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 7 6 from accumulation 1124:1135 arg1 elements 1161:1168 perinuclear endosome elements 1140:1168 perinuclear endosome elements that were accessible to transferrin 1140:1204 MAL2 depletion did not affect the internalization of these molecules but produced their accumulation in perinuclear endosome elements that were accessible to transferrin. 12370246 3 7 theme polarized 547:555 arg1 cells 563:567 polarized HepG2 cells 547:567 polarized HepG2 cells 547:567 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 6 8 theme polymeric 912:920 arg1 receptor 937:944 exogenous polymeric immunoglobulin receptor 902:944 exogenous polymeric immunoglobulin receptor 902:944 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 2 9 theme direct 420:425 arg1 route 427:431 the direct route 416:431 the direct route in MDCK cells 416:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 3 10 theme HepG2 557:561 arg1 cells 563:567 polarized HepG2 cells 547:567 polarized HepG2 cells 547:567 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 1 11 from Golgi 236:240 arg1 pathway 219:225 a direct pathway 210:225 a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface 210:336 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 6 12 theme exogenous 902:910 arg1 receptor 937:944 exogenous polymeric immunoglobulin receptor 902:944 exogenous polymeric immunoglobulin receptor 902:944 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 3 13 theme family 536:541 arg1 MAL2 502:505 MAL2 502:505 MAL2 502:505 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 3 13 theme family 536:541 arg1 member 510:515 a member 508:515 a member of the MAL protein family 508:541 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 4 14 theme localized 654:662 arg1 compartment 642:652 a compartment localized 640:662 a compartment localized beneath the subapical F-actin cytoskeleton 640:705 MAL2 resided selectively in rafts and is predominantly distributed in a compartment localized beneath the subapical F-actin cytoskeleton. 12370246 4 15 theme F-actin 686:692 arg1 cytoskeleton 694:705 the subapical F-actin cytoskeleton 672:705 the subapical F-actin cytoskeleton 672:705 MAL2 resided selectively in rafts and is predominantly distributed in a compartment localized beneath the subapical F-actin cytoskeleton. 12370246 6 16 theme glycosylphosphatidylinositol-anchored 961:997 arg1 protein 999:1005 endogenous glycosylphosphatidylinositol-anchored protein CD59 950:1010 endogenous glycosylphosphatidylinositol-anchored protein CD59 950:1010 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 2 17 theme MAL 359:361 arg1 element 387:393 an essential element 374:393 an essential element of the machinery for the direct route in MDCK cells 374:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 2 17 theme MAL 359:361 arg1 protein 363:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 1 18 theme indirect 278:285 arg1 route 287:291 an indirect route 275:291 an indirect route for targeting proteins to the apical surface 275:336 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 2 19 theme machinery 402:410 arg1 element 387:393 an essential element 374:393 an essential element of the machinery for the direct route in MDCK cells 374:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 2 19 theme machinery 402:410 arg1 protein 363:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 7 20 theme endosome 1152:1159 arg1 elements 1161:1168 perinuclear endosome elements 1140:1168 perinuclear endosome elements that were accessible to transferrin 1140:1204 MAL2 depletion did not affect the internalization of these molecules but produced their accumulation in perinuclear endosome elements that were accessible to transferrin. 12370246 2 21 theme raft-associated 343:357 arg1 element 387:393 an essential element 374:393 an essential element of the machinery for the direct route in MDCK cells 374:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 2 21 theme raft-associated 343:357 arg1 protein 363:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 6 22 theme receptor 937:944 arg1 transport 889:897 transcytotic transport 876:897 transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane 876:1033 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 7 23 theme molecules 1095:1103 arg1 internalization 1070:1084 the internalization 1066:1084 the internalization of these molecules 1066:1103 MAL2 depletion did not affect the internalization of these molecules but produced their accumulation in perinuclear endosome elements that were accessible to transferrin. 12370246 0 24 theme novel 8:12 arg1 protein 19:25 a novel raft protein 6:25 a novel raft protein of the MAL family 6:43 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 24 theme novel 8:12 arg1 MAL2 0:3 MAL2 0:3 MAL2 0:3 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 1 25 dep cells 265:269 arg1 e.g. 243:246 e.g. 243:246 e.g. 243:246 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 0 26 theme machinery 79:87 arg1 MAL2 0:3 MAL2 0:3 MAL2 0:3 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 26 theme machinery 79:87 arg1 component 62:70 an essential component 49:70 an essential component of the machinery for transcytosis in hepatoma HepG2 cells 49:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 6 27 theme apical 1019:1024 arg1 membrane 1026:1033 the apical membrane 1015:1033 the apical membrane 1015:1033 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 7 28 theme perinuclear 1140:1150 arg1 elements 1161:1168 perinuclear endosome elements 1140:1168 perinuclear endosome elements that were accessible to transferrin 1140:1204 MAL2 depletion did not affect the internalization of these molecules but produced their accumulation in perinuclear endosome elements that were accessible to transferrin. 12370246 6 29 theme transcytotic 876:887 arg1 transport 889:897 transcytotic transport 876:897 transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane 876:1033 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 9 30 from transcytosis 1352:1363 arg1 cells 1374:1378 HepG2 cells 1368:1378 HepG2 cells 1368:1378 MAL2 is therefore essential for transcytosis in HepG2 cells. 12370246 1 31 dep cells 180:184 arg1 e.g. 159:162 e.g. 159:162 e.g. 159:162 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 3 32 from characterization 482:497 arg1 cells 563:567 polarized HepG2 cells 547:567 polarized HepG2 cells 547:567 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 2 33 theme essential 377:385 arg1 element 387:393 an essential element 374:393 an essential element of the machinery for the direct route in MDCK cells 374:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 2 33 theme essential 377:385 arg1 protein 363:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein 339:369 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 2 34 theme MDCK 436:439 arg1 cells 441:445 MDCK cells 436:445 MDCK cells 436:445 The raft-associated MAL protein is an essential element of the machinery for the direct route in MDCK cells. 12370246 3 35 theme protein 528:534 arg1 family 536:541 the MAL protein family 520:541 the MAL protein family 520:541 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 5 36 theme subapical 736:744 arg1 structures 755:764 subapical endosome structures 736:764 subapical endosome structures 736:764 MAL2 greatly colocalized in subapical endosome structures with transcytosing molecules en route to the apical surface. 12370246 0 37 theme raft 14:17 arg1 protein 19:25 a novel raft protein 6:25 a novel raft protein of the MAL family 6:43 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 37 theme raft 14:17 arg1 MAL2 0:3 MAL2 0:3 MAL2 0:3 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 6 38 theme immunoglobulin 922:935 arg1 receptor 937:944 exogenous polymeric immunoglobulin receptor 902:944 exogenous polymeric immunoglobulin receptor 902:944 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 6 39 theme endogenous 950:959 arg1 protein 999:1005 endogenous glycosylphosphatidylinositol-anchored protein CD59 950:1010 endogenous glycosylphosphatidylinositol-anchored protein CD59 950:1010 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 1 40 with combination 193:203 arg1 pathway 219:225 a direct pathway 210:225 a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface 210:336 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 5 41 theme endosome 746:753 arg1 structures 755:764 subapical endosome structures 736:764 subapical endosome structures 736:764 MAL2 greatly colocalized in subapical endosome structures with transcytosing molecules en route to the apical surface. 12370246 6 42 theme MAL2 851:854 arg1 Depletion 827:835 Depletion 827:835 Depletion of endogenous MAL2 827:854 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 0 43 theme hepatoma 109:116 arg1 cells 124:128 hepatoma HepG2 cells 109:128 hepatoma HepG2 cells 109:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 1 44 theme direct 212:217 arg1 pathway 219:225 a direct pathway 210:225 a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface 210:336 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 5 45 theme apical 811:816 arg1 surface 818:824 the apical surface 807:824 the apical surface 807:824 MAL2 greatly colocalized in subapical endosome structures with transcytosing molecules en route to the apical surface. 12370246 8 46 theme Normal 1207:1212 arg1 transcytosis 1214:1225 Normal transcytosis 1207:1225 Normal transcytosis 1207:1225 Normal transcytosis persisted in cells that expressed exogenous MAL2 designed to resist the depletion treatment. 12370246 6 47 theme endogenous 840:849 arg1 MAL2 851:854 endogenous MAL2 840:854 endogenous MAL2 840:854 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 7 48 theme MAL2 1036:1039 arg1 depletion 1041:1049 MAL2 depletion 1036:1049 MAL2 depletion 1036:1049 MAL2 depletion did not affect the internalization of these molecules but produced their accumulation in perinuclear endosome elements that were accessible to transferrin. 12370246 3 49 theme functional 471:480 arg1 characterization 482:497 the functional characterization 467:497 the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells 467:567 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 0 50 from component 62:70 arg1 cells 124:128 hepatoma HepG2 cells 109:128 hepatoma HepG2 cells 109:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 1 51 used used 147:150 arg2 Transcytosis 131:142 Transcytosis 131:142 Transcytosis 131:142 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 8 52 theme depletion 1299:1307 arg1 treatment 1309:1317 the depletion treatment 1295:1317 the depletion treatment 1295:1317 Normal transcytosis persisted in cells that expressed exogenous MAL2 designed to resist the depletion treatment. 12370246 3 53 theme MAL2 502:505 arg1 characterization 482:497 the functional characterization 467:497 the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells 467:567 Herein, we present the functional characterization of MAL2, a member of the MAL protein family, in polarized HepG2 cells. 12370246 1 54 theme hepatoma 165:172 arg1 cells 180:184 hepatoma HepG2 cells 165:184 hepatoma HepG2 cells 165:184 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 0 55 theme HepG2 118:122 arg1 cells 124:128 hepatoma HepG2 cells 109:128 hepatoma HepG2 cells 109:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 1 56 theme apical 323:328 arg1 surface 330:336 the apical surface 319:336 the apical surface 319:336 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 1 57 theme HepG2 174:178 arg1 cells 180:184 hepatoma HepG2 cells 165:184 hepatoma HepG2 cells 165:184 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 0 58 theme family 38:43 arg1 protein 19:25 a novel raft protein 6:25 a novel raft protein of the MAL family 6:43 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 58 theme family 38:43 arg1 MAL2 0:3 MAL2 0:3 MAL2 0:3 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 1 59 dep alone 152:156 arg1 cells 180:184 hepatoma HepG2 cells 165:184 hepatoma HepG2 cells 165:184 Transcytosis is used alone (e.g., hepatoma HepG2 cells) or in combination with a direct pathway from the Golgi (e.g., epithelial MDCK cells) as an indirect route for targeting proteins to the apical surface. 12370246 8 60 theme exogenous 1261:1269 arg1 MAL2 1271:1274 exogenous MAL2 1261:1274 exogenous MAL2 designed to resist the depletion treatment 1261:1317 Normal transcytosis persisted in cells that expressed exogenous MAL2 designed to resist the depletion treatment. 12370246 0 61 from machinery 79:87 arg1 cells 124:128 hepatoma HepG2 cells 109:128 hepatoma HepG2 cells 109:128 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 0 62 theme MAL 34:36 arg1 family 38:43 the MAL family 30:43 the MAL family 30:43 MAL2, a novel raft protein of the MAL family, is an essential component of the machinery for transcytosis in hepatoma HepG2 cells. 12370246 4 63 theme subapical 676:684 arg1 cytoskeleton 694:705 the subapical F-actin cytoskeleton 672:705 the subapical F-actin cytoskeleton 672:705 MAL2 resided selectively in rafts and is predominantly distributed in a compartment localized beneath the subapical F-actin cytoskeleton. 12370246 6 64 theme protein 999:1005 arg1 transport 889:897 transcytotic transport 876:897 transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane 876:1033 Depletion of endogenous MAL2 drastically blocked transcytotic transport of exogenous polymeric immunoglobulin receptor and endogenous glycosylphosphatidylinositol-anchored protein CD59 to the apical membrane. 12370246 9 65 theme HepG2 1368:1372 arg1 cells 1374:1378 HepG2 cells 1368:1378 HepG2 cells 1368:1378 MAL2 is therefore essential for transcytosis in HepG2 cells. 20427278 5 0 theme APP 697:699 arg1 N- 671:672 complex N- 663:672 complex N- 663:672 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 5 0 theme APP 697:699 arg1 O-glycosylation 678:692 O-glycosylation 678:692 O-glycosylation of APP in cultured cells 678:717 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 1 1 theme disease 307:313 arg1 Abeta 337:341 Abeta 337:341 Abeta 337:341 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 1 theme disease 307:313 arg1 peptide 328:334 the Alzheimer disease amyloid beta peptide 293:334 the Alzheimer disease amyloid beta peptide (Abeta) 293:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 9 2 theme oligomeric 1427:1436 arg1 proteins 1450:1457 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 2 theme oligomeric 1427:1436 arg1 COG2 1468:1471 COG2 1468:1471 COG2 1468:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 2 theme oligomeric 1427:1436 arg1 COG1 1459:1462 COG1 1459:1462 COG1 1459:1462 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 3 theme COG 1445:1447 arg1 proteins 1450:1457 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 3 theme COG 1445:1447 arg1 COG2 1468:1471 COG2 1468:1471 COG2 1468:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 3 theme COG 1445:1447 arg1 COG1 1459:1462 COG1 1459:1462 COG1 1459:1462 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 6 4 theme APP 818:820 arg1 cleavage 822:829 APP cleavage 818:829 Abeta generation as well as APP cleavage 790:829 Additionally, TMEM59 induced APP retention in the Golgi and inhibited Abeta generation as well as APP cleavage by alpha- and beta-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. 20427278 11 5 theme APP 1696:1698 arg1 shedding 1700:1707 APP shedding 1696:1707 APP shedding 1696:1707 In summary, this study shows that TMEM59 expression modulates complex N- and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by alpha- and beta-secretase. 20427278 0 6 theme surface 72:78 arg1 expression 80:89 cell surface expression 67:89 cell surface expression 67:89 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 1 7 theme amyloid 315:321 arg1 Abeta 337:341 Abeta 337:341 Abeta 337:341 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 7 theme amyloid 315:321 arg1 peptide 328:334 the Alzheimer disease amyloid beta peptide 293:334 the Alzheimer disease amyloid beta peptide (Abeta) 293:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 7 8 theme general 1036:1042 arg1 modulation 1044:1053 a more general modulation 1029:1053 a more general modulation of Golgi glycosylation reactions 1029:1086 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 6 9 theme APP 749:751 arg1 retention 753:761 APP retention 749:761 APP retention 749:761 Additionally, TMEM59 induced APP retention in the Golgi and inhibited Abeta generation as well as APP cleavage by alpha- and beta-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. 20427278 8 10 theme general 1273:1279 arg1 function 1287:1294 the general Golgi function 1269:1294 the general Golgi function 1269:1294 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 5 11 from N- 671:672 arg1 cells 713:717 cultured cells 704:717 cultured cells 704:717 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 1 12 theme beta 323:326 arg1 Abeta 337:341 Abeta 337:341 Abeta 337:341 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 12 theme beta 323:326 arg1 peptide 328:334 the Alzheimer disease amyloid beta peptide 293:334 the Alzheimer disease amyloid beta peptide (Abeta) 293:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 5 13 theme complex 663:669 arg1 N- 671:672 complex N- 663:672 complex N- 663:672 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 1 14 theme peptide 328:334 arg1 generation 279:288 the generation 275:288 the generation of the Alzheimer disease amyloid beta peptide (Abeta) 275:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 5 15 gly O-glycosylation 678:692 arg1 APP 697:699 APP 697:699 APP 697:699 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 5 15 gly O-glycosylation 678:692 arg1 cells 713:717 cultured cells 704:717 cultured cells 704:717 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 3 16 theme transmembrane 488:500 arg1 protein 479:485 a novel protein 471:485 a novel protein 471:485 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 3 16 theme transmembrane 488:500 arg1 protein 502:508 transmembrane protein 59 488:511 transmembrane protein 59 (TMEM59) 488:520 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 3 16 theme transmembrane 488:500 arg1 TMEM59 514:519 TMEM59 514:519 TMEM59 514:519 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 8 17 theme alpha 1220:1224 arg1 shedding 1186:1193 shedding 1186:1193 shedding of tumor necrosis factor alpha 1186:1224 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 11 18 theme TMEM59 1604:1609 arg1 expression 1611:1620 TMEM59 expression 1604:1620 TMEM59 expression 1604:1620 In summary, this study shows that TMEM59 expression modulates complex N- and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by alpha- and beta-secretase. 20427278 8 19 theme factor 1213:1218 arg1 alpha 1220:1224 tumor necrosis factor alpha 1198:1224 tumor necrosis factor alpha 1198:1224 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 1 20 from event 266:270 arg1 generation 279:288 the generation 275:288 the generation of the Alzheimer disease amyloid beta peptide (Abeta) 275:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 3 21 theme novel 473:477 arg1 modulator 532:540 a new modulator 526:540 a new modulator of APP shedding 526:556 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 3 21 theme novel 473:477 arg1 protein 479:485 a novel protein 471:485 a novel protein 471:485 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 3 21 theme novel 473:477 arg1 protein 502:508 transmembrane protein 59 488:511 transmembrane protein 59 (TMEM59) 488:520 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 10 22 theme enzymes 1561:1567 arg1 activity 1529:1536 activity 1529:1536 activity 1529:1536 Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. 20427278 10 22 theme enzymes 1561:1567 arg1 localization 1512:1523 normal localization 1505:1523 normal localization 1505:1523 Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. 20427278 9 23 theme APP 1337:1339 arg1 glycosylation 1341:1353 APP glycosylation 1337:1353 APP glycosylation 1337:1353 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 7 24 theme Golgi 1058:1062 arg1 reactions 1078:1086 Golgi glycosylation reactions 1058:1086 Golgi glycosylation reactions 1058:1086 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 9 25 located observed 1391:1398 arg1 cells 1403:1407 cells 1403:1407 cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2 1403:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 25 located observed 1391:1398 arg2 one 1387:1389 the one 1383:1389 the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2 1383:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 1 26 theme amyloid 167:173 arg1 APP 194:196 APP 194:196 APP 194:196 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 26 theme amyloid 167:173 arg1 protein 185:191 amyloid precursor protein 167:191 the amyloid precursor protein (APP) 163:197 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 0 27 theme novel 4:8 arg1 protein 19:25 The novel membrane protein TMEM59 0:32 The novel membrane protein TMEM59 0:32 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 2 28 theme Abeta 433:437 arg1 generation 439:448 Abeta generation 433:448 Abeta generation 433:448 At present, little is known about the cellular mechanisms that control APP shedding and Abeta generation. 20427278 10 29 theme normal 1505:1510 arg1 localization 1512:1523 normal localization 1505:1523 normal localization 1505:1523 Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. 20427278 6 30 theme plasma 889:894 arg1 membrane 896:903 the plasma membrane 885:903 the plasma membrane 885:903 Additionally, TMEM59 induced APP retention in the Golgi and inhibited Abeta generation as well as APP cleavage by alpha- and beta-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. 20427278 1 31 theme precursor 175:183 arg1 APP 194:196 APP 194:196 APP 194:196 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 31 theme precursor 175:183 arg1 protein 185:191 amyloid precursor protein 167:191 the amyloid precursor protein (APP) 163:197 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 5 32 theme TMEM59 633:638 arg1 transfection 640:651 TMEM59 transfection 633:651 TMEM59 transfection 633:651 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 1 33 theme key 251:253 arg1 event 266:270 a key regulatory event 249:270 a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta) 249:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 33 theme key 251:253 arg1 shedding 151:158 Ectodomain shedding 140:158 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase 140:244 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 7 34 theme prion 1003:1007 arg1 protein 1009:1015 the prion protein 999:1015 the prion protein 999:1015 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 7 35 theme protein 1009:1015 arg1 N-glycosylation 980:994 the complex N-glycosylation 968:994 the complex N-glycosylation of the prion protein 968:1015 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 1 36 theme protein 185:191 arg1 event 266:270 a key regulatory event 249:270 a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta) 249:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 36 theme protein 185:191 arg1 shedding 151:158 Ectodomain shedding 140:158 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase 140:244 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 2 37 theme cellular 383:390 arg1 mechanisms 392:401 the cellular mechanisms 379:401 the cellular mechanisms that control APP shedding and Abeta generation 379:448 At present, little is known about the cellular mechanisms that control APP shedding and Abeta generation. 20427278 1 38 theme regulatory 255:264 arg1 event 266:270 a key regulatory event 249:270 a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta) 249:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 38 theme regulatory 255:264 arg1 shedding 151:158 Ectodomain shedding 140:158 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase 140:244 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 0 39 theme amyloid 113:119 arg1 protein 131:137 the amyloid precursor protein 109:137 the amyloid precursor protein 109:137 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 0 40 theme membrane 10:17 arg1 protein 19:25 The novel membrane protein TMEM59 0:32 The novel membrane protein TMEM59 0:32 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 4 41 dep expressed 597:605 arg1 Golgi-localized 608:622 Golgi-localized 608:622 Golgi-localized 608:622 TMEM59 was found to be a ubiquitously expressed, Golgi-localized protein. 20427278 1 42 dep proteases 210:218 arg1 proteases 210:218 the two proteases alpha- and beta-secretase 202:244 the two proteases alpha- and beta-secretase 202:244 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 42 dep proteases 210:218 arg1 beta-secretase 231:244 beta-secretase 231:244 beta-secretase 231:244 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 42 dep proteases 210:218 arg1 alpha- 220:225 alpha- 220:225 alpha- 220:225 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 9 43 theme Golgi 1438:1442 arg1 proteins 1450:1457 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 43 theme Golgi 1438:1442 arg1 COG2 1468:1471 COG2 1468:1471 COG2 1468:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 43 theme Golgi 1438:1442 arg1 COG1 1459:1462 COG1 1459:1462 COG1 1459:1462 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 5 44 theme cultured 704:711 arg1 cells 713:717 cultured cells 704:717 cultured cells 704:717 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 0 45 theme protein 131:137 arg1 expression 80:89 cell surface expression 67:89 cell surface expression 67:89 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 0 45 theme protein 131:137 arg1 secretion 96:104 secretion 96:104 secretion of the amyloid precursor protein 96:137 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 0 45 theme protein 131:137 arg1 glycosylation 52:64 complex glycosylation 44:64 complex glycosylation 44:64 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 6 46 theme Abeta 790:794 arg1 generation 796:805 Abeta generation 790:805 Abeta generation as well as APP cleavage 790:829 Additionally, TMEM59 induced APP retention in the Golgi and inhibited Abeta generation as well as APP cleavage by alpha- and beta-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. 20427278 10 47 theme glycosylation 1547:1559 arg1 enzymes 1561:1567 Golgi glycosylation enzymes 1541:1567 Golgi glycosylation enzymes 1541:1567 Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. 20427278 8 48 theme tumor 1198:1202 arg1 alpha 1220:1224 tumor necrosis factor alpha 1198:1224 tumor necrosis factor alpha 1198:1224 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 7 49 theme glycosylation 1064:1076 arg1 reactions 1078:1086 Golgi glycosylation reactions 1058:1086 Golgi glycosylation reactions 1058:1086 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 0 50 theme precursor 121:129 arg1 protein 131:137 the amyloid precursor protein 109:137 the amyloid precursor protein 109:137 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 3 51 theme shedding 549:556 arg1 modulator 532:540 a new modulator 526:540 a new modulator of APP shedding 526:556 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 3 51 theme shedding 549:556 arg1 protein 479:485 a novel protein 471:485 a novel protein 471:485 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 7 52 theme reactions 1078:1086 arg1 modulation 1044:1053 a more general modulation 1029:1053 a more general modulation of Golgi glycosylation reactions 1029:1086 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 2 53 theme APP 416:418 arg1 shedding 420:427 APP shedding 416:427 APP shedding 416:427 At present, little is known about the cellular mechanisms that control APP shedding and Abeta generation. 20427278 11 54 theme cellular 1742:1749 arg1 compartments 1751:1762 the cellular compartments 1738:1762 the cellular compartments 1738:1762 In summary, this study shows that TMEM59 expression modulates complex N- and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by alpha- and beta-secretase. 20427278 8 55 theme Golgi 1281:1285 arg1 function 1287:1294 the general Golgi function 1269:1294 the general Golgi function 1269:1294 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 7 56 theme complex 972:978 arg1 N-glycosylation 980:994 the complex N-glycosylation 968:994 the complex N-glycosylation of the prion protein 968:1015 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 0 57 theme complex 44:50 arg1 glycosylation 52:64 complex glycosylation 44:64 complex glycosylation 44:64 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 11 58 theme APP 1731:1733 arg1 access 1721:1726 access 1721:1726 access of APP to the cellular compartments, where it is normally cleaved by alpha- and beta-secretase 1721:1821 In summary, this study shows that TMEM59 expression modulates complex N- and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by alpha- and beta-secretase. 20427278 8 59 theme necrosis 1204:1211 arg1 alpha 1220:1224 tumor necrosis factor alpha 1198:1224 tumor necrosis factor alpha 1198:1224 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 10 60 theme Golgi 1541:1545 arg1 enzymes 1561:1567 Golgi glycosylation enzymes 1541:1567 Golgi glycosylation enzymes 1541:1567 Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. 20427278 6 61 theme beta-secretase 845:858 arg1 cleavage 860:867 beta-secretase cleavage 845:867 beta-secretase cleavage 845:867 Additionally, TMEM59 induced APP retention in the Golgi and inhibited Abeta generation as well as APP cleavage by alpha- and beta-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. 20427278 9 62 from phenotype 1301:1309 arg1 shedding 1359:1366 shedding 1359:1366 shedding 1359:1366 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 62 from phenotype 1301:1309 arg1 glycosylation 1341:1353 APP glycosylation 1337:1353 APP glycosylation 1337:1353 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 63 theme conserved 1417:1425 arg1 proteins 1450:1457 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 63 theme conserved 1417:1425 arg1 COG2 1468:1471 COG2 1468:1471 COG2 1468:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 63 theme conserved 1417:1425 arg1 COG1 1459:1462 COG1 1459:1462 COG1 1459:1462 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 5 64 from O-glycosylation 678:692 arg1 cells 713:717 cultured cells 704:717 cultured cells 704:717 TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. 20427278 3 65 theme APP 545:547 arg1 shedding 549:556 APP shedding 545:556 APP shedding 545:556 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 7 66 gly N-glycosylation 980:994 arg1 protein 1009:1015 the prion protein 999:1015 the prion protein 999:1015 Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. 20427278 9 67 dep proteins 1450:1457 arg1 proteins 1450:1457 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 conserved oligomeric Golgi (COG) proteins COG1 and COG2 1417:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 67 dep proteins 1450:1457 arg1 COG2 1468:1471 COG2 1468:1471 COG2 1468:1471 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 67 dep proteins 1450:1457 arg1 COG1 1459:1462 COG1 1459:1462 COG1 1459:1462 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 0 68 theme cell 67:70 arg1 expression 80:89 cell surface expression 67:89 cell surface expression 67:89 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 11 69 theme complex 1632:1638 arg1 N- 1640:1641 complex N- 1632:1641 complex N- 1632:1641 In summary, this study shows that TMEM59 expression modulates complex N- and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by alpha- and beta-secretase. 20427278 3 70 theme new 528:530 arg1 modulator 532:540 a new modulator 526:540 a new modulator of APP shedding 526:556 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 3 70 theme new 528:530 arg1 protein 479:485 a novel protein 471:485 a novel protein 471:485 Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. 20427278 4 71 theme expressed 597:605 arg1 TMEM59 559:564 TMEM59 559:564 TMEM59 559:564 TMEM59 was found to be a ubiquitously expressed, Golgi-localized protein. 20427278 4 71 theme expressed 597:605 arg1 protein 624:630 a ubiquitously expressed, Golgi-localized protein 582:630 a ubiquitously expressed, Golgi-localized protein 582:630 TMEM59 was found to be a ubiquitously expressed, Golgi-localized protein. 20427278 8 72 theme proteins 1149:1156 arg1 secretion 1128:1136 the secretion 1124:1136 the secretion of soluble proteins 1124:1156 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 8 72 theme proteins 1149:1156 arg1 alpha-secretase 1165:1179 the alpha-secretase 1161:1179 the alpha-secretase like shedding of tumor necrosis factor alpha 1161:1224 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 8 73 theme soluble 1141:1147 arg1 proteins 1149:1156 soluble proteins 1141:1156 soluble proteins 1141:1156 Importantly, TMEM59 did not affect the secretion of soluble proteins or the alpha-secretase like shedding of tumor necrosis factor alpha, demonstrating that TMEM59 did not disturb the general Golgi function. 20427278 1 74 theme Alzheimer 297:305 arg1 Abeta 337:341 Abeta 337:341 Abeta 337:341 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 74 theme Alzheimer 297:305 arg1 peptide 328:334 the Alzheimer disease amyloid beta peptide 293:334 the Alzheimer disease amyloid beta peptide (Abeta) 293:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 0 75 gly glycosylation 52:64 arg1 protein 131:137 the amyloid precursor protein 109:137 the amyloid precursor protein 109:137 The novel membrane protein TMEM59 modulates complex glycosylation, cell surface expression, and secretion of the amyloid precursor protein. 20427278 1 76 theme Ectodomain 140:149 arg1 event 266:270 a key regulatory event 249:270 a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta) 249:342 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 1 76 theme Ectodomain 140:149 arg1 shedding 151:158 Ectodomain shedding 140:158 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase 140:244 Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases alpha- and beta-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid beta peptide (Abeta). 20427278 9 77 theme transfection 1321:1332 arg1 similar 1372:1378 similar 1372:1378 similar 1372:1378 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 20427278 9 77 theme transfection 1321:1332 arg1 phenotype 1301:1309 The phenotype 1297:1309 The phenotype of TMEM59 transfection on APP glycosylation and shedding 1297:1366 The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. 23139753 5 0 theme cysteine 661:668 arg1 proteases 670:678 the serine, aspartyl and cysteine proteases 636:678 proteases 670:678 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 1 1 theme sperm 145:149 arg1 maturation 131:140 maturation 131:140 maturation of sperm 131:149 Epididymal proteins represent the factors necessary for maturation of sperm and play a crucial role in sperm maturation. 23139753 0 2 theme protease 55:62 arg1 inhibitor 64:72 a novel cross-class protease inhibitor 35:72 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 0:73 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 4 3 theme acid 437:440 arg1 HE-4 417:420 HE-4 417:420 HE-4 417:420 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 4 3 theme acid 437:440 arg1 polypeptide 447:457 a 124 amino acid long polypeptide 425:457 a 124 amino acid long polypeptide with two WFDC domains 425:479 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 8 4 theme temperature 945:955 arg1 range 929:933 a wide range 922:933 a wide range of pH and temperature 922:955 Moreover, HE-4 is N-glycosylated and highly stable on a wide range of pH and temperature. 23139753 6 5 with binding 782:788 arg1 proteases 805:813 all these proteases 795:813 all these proteases 795:813 Using SPR we have also observed that HE-4 shows a significant binding with all these proteases. 23139753 8 6 gly N-glycosylated 886:899 arg1 HE-4 878:881 HE-4 878:881 HE-4 878:881 Moreover, HE-4 is N-glycosylated and highly stable on a wide range of pH and temperature. 23139753 4 7 theme amino 431:435 arg1 HE-4 417:420 HE-4 417:420 HE-4 417:420 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 4 7 theme amino 431:435 arg1 polypeptide 447:457 a 124 amino acid long polypeptide 425:457 a 124 amino acid long polypeptide with two WFDC domains 425:479 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 5 8 theme disulfide-bonded 550:565 arg1 HE-4 501:504 HE-4 501:504 HE-4 501:504 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 5 8 theme disulfide-bonded 550:565 arg1 homo-trimer 567:577 a disulfide-bonded homo-trimer 548:577 a disulfide-bonded homo-trimer 548:577 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 9 9 theme microbial 1071:1079 arg1 factors 1091:1097 microbial virulence factors 1071:1097 microbial virulence factors of proteolytic nature 1071:1119 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 2 10 theme acidic 245:250 arg1 family 287:292 whey acidic protein four-disulfide core (WFDC) family 240:292 whey acidic protein four-disulfide core (WFDC) family with no known function 240:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 7 11 theme Disulfide 816:824 arg1 linkages 826:833 Disulfide linkages 816:833 Disulfide linkages 816:833 Disulfide linkages are essential for this activity. 23139753 2 12 theme whey 240:243 arg1 family 287:292 whey acidic protein four-disulfide core (WFDC) family 240:292 whey acidic protein four-disulfide core (WFDC) family with no known function 240:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 9 13 theme virulence 1081:1089 arg1 factors 1091:1097 microbial virulence factors 1071:1097 microbial virulence factors of proteolytic nature 1071:1119 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 2 14 theme known 302:306 arg1 function 308:315 no known function 299:315 no known function 299:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 5 15 theme serine 640:645 arg1 proteases 670:678 the serine, aspartyl and cysteine proteases 636:678 proteases 670:678 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 8 16 theme pH 938:939 arg1 range 929:933 a wide range 922:933 a wide range of pH and temperature 922:955 Moreover, HE-4 is N-glycosylated and highly stable on a wide range of pH and temperature. 23139753 3 17 theme cystine 400:406 arg1 residue 408:414 eight conserved cystine residue 384:414 eight conserved cystine residue 384:414 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 3 18 theme amino 367:371 arg1 acids 373:377 50 amino acids 364:377 50 amino acids 364:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 8 19 theme wide 924:927 arg1 range 929:933 a wide range 922:933 a wide range of pH and temperature 922:955 Moreover, HE-4 is N-glycosylated and highly stable on a wide range of pH and temperature. 23139753 6 20 theme significant 770:780 arg1 binding 782:788 a significant binding 768:788 a significant binding with all these proteases 768:813 Using SPR we have also observed that HE-4 shows a significant binding with all these proteases. 23139753 5 21 theme human 525:529 arg1 fluid 539:543 the human seminal fluid 521:543 the human seminal fluid 521:543 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 3 22 theme acids 373:377 arg1 acids 373:377 50 amino acids 364:377 50 amino acids 364:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 3 22 theme acids 373:377 arg1 domain 354:359 a conserved WFDC domain 337:359 a conserved WFDC domain of 50 amino acids 337:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 0 23 theme epididymis 6:15 arg1 protein-4 17:25 Human epididymis protein-4 0:25 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 0:73 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 0 23 theme epididymis 6:15 arg1 HE-4 28:31 HE-4 28:31 HE-4 28:31 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 5 24 dep inhibitor 609:617 arg1 inhibits 619:626 inhibits 619:626 inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate 619:717 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 5 25 theme seminal 531:537 arg1 fluid 539:543 the human seminal fluid 521:543 the human seminal fluid 521:543 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 5 26 theme aspartyl 648:655 arg1 proteases 670:678 the serine, aspartyl and cysteine proteases 636:678 proteases 670:678 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 9 27 theme nature 1114:1119 arg1 factors 1091:1097 microbial virulence factors 1071:1097 microbial virulence factors of proteolytic nature 1071:1119 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 1 28 theme crucial 162:168 arg1 role 170:173 a crucial role 160:173 a crucial role 160:173 Epididymal proteins represent the factors necessary for maturation of sperm and play a crucial role in sperm maturation. 23139753 0 29 theme Human 0:4 arg1 protein-4 17:25 Human epididymis protein-4 0:25 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 0:73 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 0 29 theme Human 0:4 arg1 HE-4 28:31 HE-4 28:31 HE-4 28:31 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 3 30 contain has 333:335 arg1 protein 325:331 A WFDC protein 318:331 A WFDC protein 318:331 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 3 30 contain has 333:335 arg2 domain 354:359 a conserved WFDC domain 337:359 a conserved WFDC domain of 50 amino acids 337:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 3 30 contain has 333:335 arg2 acids 373:377 50 amino acids 364:377 50 amino acids 364:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 1 31 theme Epididymal 75:84 arg1 proteins 86:93 Epididymal proteins 75:93 Epididymal proteins 75:93 Epididymal proteins represent the factors necessary for maturation of sperm and play a crucial role in sperm maturation. 23139753 4 32 theme WFDC 468:471 arg1 domains 473:479 two WFDC domains 464:479 two WFDC domains 464:479 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 5 33 theme protease 600:607 arg1 inhibitor 609:617 a cross-class protease inhibitor 586:617 a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate 586:717 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 2 34 theme four-disulfide 260:273 arg1 family 287:292 whey acidic protein four-disulfide core (WFDC) family 240:292 whey acidic protein four-disulfide core (WFDC) family with no known function 240:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 3 35 theme conserved 390:398 arg1 residue 408:414 eight conserved cystine residue 384:414 eight conserved cystine residue 384:414 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 2 36 theme core 275:278 arg1 family 287:292 whey acidic protein four-disulfide core (WFDC) family 240:292 whey acidic protein four-disulfide core (WFDC) family with no known function 240:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 2 37 theme protein 252:258 arg1 family 287:292 whey acidic protein four-disulfide core (WFDC) family 240:292 whey acidic protein four-disulfide core (WFDC) family with no known function 240:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 1 38 theme sperm 178:182 arg1 maturation 184:193 sperm maturation 178:193 sperm maturation 178:193 Epididymal proteins represent the factors necessary for maturation of sperm and play a crucial role in sperm maturation. 23139753 3 39 theme WFDC 349:352 arg1 acids 373:377 50 amino acids 364:377 50 amino acids 364:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 3 39 theme WFDC 349:352 arg1 domain 354:359 a conserved WFDC domain 337:359 a conserved WFDC domain of 50 amino acids 337:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 2 40 theme epididymal 205:214 arg1 HE-4 196:199 HE-4 196:199 HE-4 196:199 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 2 40 theme epididymal 205:214 arg1 protein 216:222 an epididymal protein 202:222 an epididymal protein 202:222 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 4 41 theme long 442:445 arg1 HE-4 417:420 HE-4 417:420 HE-4 417:420 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 4 41 theme long 442:445 arg1 polypeptide 447:457 a 124 amino acid long polypeptide 425:457 a 124 amino acid long polypeptide with two WFDC domains 425:479 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 2 42 with family 287:292 arg1 function 308:315 no known function 299:315 no known function 299:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 9 43 theme cross-class 1002:1012 arg1 inhibitor 1023:1031 a cross-class protease inhibitor 1000:1031 a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature 1000:1119 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 9 43 theme cross-class 1002:1012 arg1 HE-4 992:995 HE-4 992:995 HE-4 992:995 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 3 44 theme conserved 339:347 arg1 acids 373:377 50 amino acids 364:377 50 amino acids 364:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 3 44 theme conserved 339:347 arg1 domain 354:359 a conserved WFDC domain 337:359 a conserved WFDC domain of 50 amino acids 337:377 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 9 45 theme protease 1014:1021 arg1 inhibitor 1023:1031 a cross-class protease inhibitor 1000:1031 a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature 1000:1119 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 9 45 theme protease 1014:1021 arg1 HE-4 992:995 HE-4 992:995 HE-4 992:995 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 2 46 theme family 287:292 arg1 member 230:235 a member 228:235 a member of whey acidic protein four-disulfide core (WFDC) family with no known function 228:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 2 46 theme family 287:292 arg1 HE-4 196:199 HE-4 196:199 HE-4 196:199 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 1 47 theme necessary 117:125 arg1 factors 109:115 the factors 105:115 the factors necessary for maturation of sperm 105:149 Epididymal proteins represent the factors necessary for maturation of sperm and play a crucial role in sperm maturation. 23139753 4 48 with polypeptide 447:457 arg1 domains 473:479 two WFDC domains 464:479 two WFDC domains 464:479 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 5 49 theme cross-class 588:598 arg1 inhibitor 609:617 a cross-class protease inhibitor 586:617 a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate 586:717 Here, we show that HE-4 is secreted in the human seminal fluid as a disulfide-bonded homo-trimer and is a cross-class protease inhibitor inhibits some of the serine, aspartyl and cysteine proteases tested using hemoglobin as a substrate. 23139753 0 50 dep protein-4 17:25 arg1 inhibitor 64:72 a novel cross-class protease inhibitor 35:72 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 0:73 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 2 51 theme WFDC 281:284 arg1 family 287:292 whey acidic protein four-disulfide core (WFDC) family 240:292 whey acidic protein four-disulfide core (WFDC) family with no known function 240:315 HE-4, an epididymal protein, is a member of whey acidic protein four-disulfide core (WFDC) family with no known function. 23139753 0 52 theme cross-class 43:53 arg1 inhibitor 64:72 a novel cross-class protease inhibitor 35:72 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 0:73 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 3 53 dep conserved 390:398 arg1 eight 384:388 eight 384:388 eight 384:388 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 9 54 theme proteolytic 1102:1112 arg1 nature 1114:1119 proteolytic nature 1102:1119 proteolytic nature 1102:1119 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 0 55 theme novel 37:41 arg1 inhibitor 64:72 a novel cross-class protease inhibitor 35:72 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 0:73 Human epididymis protein-4 (HE-4): a novel cross-class protease inhibitor. 23139753 4 56 theme 124 427:429 arg1 HE-4 417:420 HE-4 417:420 HE-4 417:420 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 4 56 theme 124 427:429 arg1 polypeptide 447:457 a 124 amino acid long polypeptide 425:457 a 124 amino acid long polypeptide with two WFDC domains 425:479 HE-4 is a 124 amino acid long polypeptide with two WFDC domains. 23139753 3 57 theme WFDC 320:323 arg1 protein 325:331 A WFDC protein 318:331 A WFDC protein 318:331 A WFDC protein has a conserved WFDC domain of 50 amino acids with eight conserved cystine residue. 23139753 9 58 dep suggests 978:985 arg1 Taken 958:962 Taken 958:962 Taken together 958:971 Taken together this suggests that HE-4 is a cross-class protease inhibitor which might confer protection against microbial virulence factors of proteolytic nature. 23139753 6 59 dep observed 743:750 arg1 Using 720:724 Using 720:724 Using SPR 720:728 Using SPR we have also observed that HE-4 shows a significant binding with all these proteases. 29632068 8 0 theme cognate 1427:1433 arg1 acyl-CoAs 1435:1443 their cognate acyl-CoAs 1421:1443 their cognate acyl-CoAs 1421:1443 Additional data supported the notion that substitutions of these 11 residues alter the specificities of the CerS toward their cognate acyl-CoAs. 29632068 6 1 dep generated 1101:1109 arg1 such 1088:1091 such 1088:1091 such 1088:1091 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 5 2 theme 11-residue 767:776 arg1 sequence 778:785 an 11-residue sequence 764:785 an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS 764:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 3 from sequence 778:785 arg1 loop 792:795 a loop 790:795 a loop located between the last two putative transmembrane domains (TMDs) of the CerS 790:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 3 4 theme CerS 474:477 arg1 regions 479:485 the CerS regions 470:485 the CerS regions that determine specificity toward different acyl-CoA substrates 470:549 Little information is available about the CerS regions that determine specificity toward different acyl-CoA substrates. 29632068 9 5 theme TMD 1669:1671 arg1 flexibility 1673:1683 TMD flexibility 1669:1683 TMD flexibility 1669:1683 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 6 6 theme other 1123:1127 arg1 ceramides 1129:1137 other ceramides 1123:1137 other ceramides 1123:1137 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 4 7 theme substrate 584:592 arg1 specificity 594:604 substrate specificity 584:604 substrate specificity 584:604 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 8 8 theme Additional 1301:1310 arg1 data 1312:1315 Additional data 1301:1315 Additional data 1301:1315 Additional data supported the notion that substitutions of these 11 residues alter the specificities of the CerS toward their cognate acyl-CoAs. 29632068 2 9 theme ceramide 281:288 arg1 activities 249:258 the biological activities 234:258 the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids 234:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 3 10 theme different 521:529 arg1 substrates 540:549 different acyl-CoA substrates 521:549 different acyl-CoA substrates 521:549 Little information is available about the CerS regions that determine specificity toward different acyl-CoA substrates. 29632068 4 11 from domain 664:669 arg1 region 619:624 a region 617:624 a region of ∼150 residues in the Tram-Lag-CLN8 domain 617:669 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 2 12 theme sphingolipids 311:323 arg1 activities 249:258 the biological activities 234:258 the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids 234:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 5 13 theme narrow 737:742 arg1 specificity 744:754 narrow specificity 737:754 narrow specificity 737:754 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 9 14 contain have 1623:1626 arg1 CerS 1586:1589 the CerS 1582:1589 the CerS acting on shorter acyl-CoAs 1582:1617 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 9 14 contain have 1623:1626 arg2 loop 1652:1655 a longer, more flexible loop 1628:1655 a longer, more flexible loop 1628:1655 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 5 15 theme putative 826:833 arg1 TMDs 858:861 TMDs 858:861 TMDs 858:861 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 15 theme putative 826:833 arg1 domains 849:855 the last two putative transmembrane domains 813:855 the last two putative transmembrane domains (TMDs) of the CerS 813:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 15 theme putative 826:833 arg1 CerS 871:874 the CerS 867:874 the CerS 867:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 9 16 dep longer 1630:1635 arg1 flexible 1643:1650 flexible 1643:1650 flexible 1643:1650 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 9 17 from conformation 1543:1554 arg1 membrane 1563:1570 the membrane 1559:1570 the membrane 1559:1570 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 2 18 theme sphingolipid 267:278 arg1 ceramide 281:288 the sphingolipid, ceramide 263:288 ceramide 281:288 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 5 19 theme located 797:803 arg1 loop 792:795 a loop 790:795 a loop located between the last two putative transmembrane domains (TMDs) of the CerS 790:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 7 20 theme chimeric 1152:1159 arg1 CerS4 1170:1174 CerS4 1170:1174 CerS4(291-301→CerS2) 1170:1189 Moreover, a chimeric protein, CerS4(291-301→CerS2), based on CerS4 (which normally generates C18-C22 ceramides) displayed significant activity toward C24:1-CoA. 29632068 7 20 theme chimeric 1152:1159 arg1 protein 1161:1167 a chimeric protein 1150:1167 a chimeric protein 1150:1167 Moreover, a chimeric protein, CerS4(291-301→CerS2), based on CerS4 (which normally generates C18-C22 ceramides) displayed significant activity toward C24:1-CoA. 29632068 2 21 theme complex 303:309 arg1 sphingolipids 311:323 more complex sphingolipids 298:323 more complex sphingolipids 298:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 9 22 theme longer 1630:1635 arg1 loop 1652:1655 a longer, more flexible loop 1628:1655 a longer, more flexible loop 1628:1655 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 7 23 theme C18-C22 1233:1239 arg1 ceramides 1241:1249 C18-C22 ceramides 1233:1249 C18-C22 ceramides 1233:1249 Moreover, a chimeric protein, CerS4(291-301→CerS2), based on CerS4 (which normally generates C18-C22 ceramides) displayed significant activity toward C24:1-CoA. 29632068 2 24 theme synthases 414:422 arg1 family 391:396 a family 389:396 a family of six ceramide synthases (CerS) 389:429 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 1 25 theme large 91:95 arg1 complexity 108:117 large structural complexity 91:117 large structural complexity 91:117 Lipids display large structural complexity, with ∼40,000 different lipids identified to date, ∼4000 of which are sphingolipids. 29632068 4 26 from residues 634:641 arg1 domain 664:669 the Tram-Lag-CLN8 domain 646:669 the Tram-Lag-CLN8 domain 646:669 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 4 27 theme Tram-Lag-CLN8 650:662 arg1 domain 664:669 the Tram-Lag-CLN8 domain 646:669 the Tram-Lag-CLN8 domain 646:669 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 2 28 theme ceramide 405:412 arg1 synthases 414:422 six ceramide synthases 401:422 six ceramide synthases (CerS) 401:429 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 2 28 theme ceramide 405:412 arg1 CerS 425:428 CerS 425:428 CerS 425:428 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 8 29 theme residues 1369:1376 arg1 substitutions 1343:1355 substitutions 1343:1355 substitutions of these 11 residues 1343:1376 Additional data supported the notion that substitutions of these 11 residues alter the specificities of the CerS toward their cognate acyl-CoAs. 29632068 9 30 theme shorter 1601:1607 arg1 acyl-CoAs 1609:1617 shorter acyl-CoAs 1601:1617 shorter acyl-CoAs 1601:1617 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 3 31 theme acyl-CoA 531:538 arg1 substrates 540:549 different acyl-CoA substrates 521:549 different acyl-CoA substrates 521:549 Little information is available about the CerS regions that determine specificity toward different acyl-CoA substrates. 29632068 5 32 theme last 817:820 arg1 TMDs 858:861 TMDs 858:861 TMDs 858:861 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 32 theme last 817:820 arg1 domains 849:855 the last two putative transmembrane domains 813:855 the last two putative transmembrane domains (TMDs) of the CerS 813:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 32 theme last 817:820 arg1 CerS 871:874 the CerS 867:874 the CerS 867:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 33 theme biochemical 708:718 arg1 analyses 720:727 biochemical analyses 708:727 biochemical analyses 708:727 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 10 34 theme acyl-CoA 1758:1765 arg1 specificity 1767:1777 the acyl-CoA specificity 1754:1777 the acyl-CoA specificity of CerS 1754:1785 In summary, we have identified an 11-residue region that determines the acyl-CoA specificity of CerS. 29632068 2 35 theme chain 341:345 arg1 length 347:352 their N-acyl chain length 328:352 their N-acyl chain length 328:352 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 2 35 theme chain 341:345 arg1 factor 215:220 A critical factor 204:220 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids 204:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 3 36 theme Little 432:437 arg1 information 439:449 Little information 432:449 Little information 432:449 Little information is available about the CerS regions that determine specificity toward different acyl-CoA substrates. 29632068 0 37 theme chain 35:39 arg1 specificity 41:51 the acyl chain specificity 26:51 the acyl chain specificity of ceramide synthases 26:73 Eleven residues determine the acyl chain specificity of ceramide synthases. 29632068 5 38 theme transmembrane 835:847 arg1 TMDs 858:861 TMDs 858:861 TMDs 858:861 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 38 theme transmembrane 835:847 arg1 domains 849:855 the last two putative transmembrane domains 813:855 the last two putative transmembrane domains (TMDs) of the CerS 813:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 38 theme transmembrane 835:847 arg1 CerS 871:874 the CerS 867:874 the CerS 867:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 4 39 from region 619:624 arg1 domain 664:669 the Tram-Lag-CLN8 domain 646:669 the Tram-Lag-CLN8 domain 646:669 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 4 40 theme residues 634:641 arg1 region 619:624 a region 617:624 a region of ∼150 residues in the Tram-Lag-CLN8 domain 617:669 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 0 41 theme acyl 30:33 arg1 specificity 41:51 the acyl chain specificity 26:51 the acyl chain specificity of ceramide synthases 26:73 Eleven residues determine the acyl chain specificity of ceramide synthases. 29632068 4 42 theme ∼150 629:632 arg1 residues 634:641 ∼150 residues 629:641 ∼150 residues in the Tram-Lag-CLN8 domain 629:669 We previously demonstrated that substrate specificity resides in a region of ∼150 residues in the Tram-Lag-CLN8 domain. 29632068 2 43 theme N-acyl 334:339 arg1 length 347:352 their N-acyl chain length 328:352 their N-acyl chain length 328:352 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 2 43 theme N-acyl 334:339 arg1 factor 215:220 A critical factor 204:220 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids 204:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 6 44 theme chimeric 898:905 arg1 CerS5 916:920 CerS5 916:920 CerS5(299-309→CerS2) 916:935 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 6 44 theme chimeric 898:905 arg1 protein 907:913 a chimeric protein 896:913 a chimeric protein 896:913 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 8 45 theme CerS 1409:1412 arg1 specificities 1388:1400 the specificities 1384:1400 the specificities of the CerS toward their cognate acyl-CoAs 1384:1443 Additional data supported the notion that substitutions of these 11 residues alter the specificities of the CerS toward their cognate acyl-CoAs. 29632068 2 46 theme critical 206:213 arg1 length 347:352 their N-acyl chain length 328:352 their N-acyl chain length 328:352 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 2 46 theme critical 206:213 arg1 factor 215:220 A critical factor 204:220 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids 204:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 9 47 theme adjacent 1505:1512 arg1 TMDs 1514:1517 adjacent TMDs 1505:1517 adjacent TMDs 1505:1517 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 6 48 theme protein 907:913 arg1 specificity 881:891 The specificity 877:891 The specificity 877:891 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 5 49 theme site-directed 678:690 arg1 mutagenesis 692:702 site-directed mutagenesis 678:702 site-directed mutagenesis 678:702 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 10 50 theme CerS 1782:1785 arg1 specificity 1767:1777 the acyl-CoA specificity 1754:1777 the acyl-CoA specificity of CerS 1754:1785 In summary, we have identified an 11-residue region that determines the acyl-CoA specificity of CerS. 29632068 9 51 theme open 1538:1541 arg1 conformation 1543:1554 a more open conformation 1531:1554 a more open conformation in the membrane 1531:1570 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 1 52 theme structural 97:106 arg1 complexity 108:117 large structural complexity 91:117 large structural complexity 91:117 Lipids display large structural complexity, with ∼40,000 different lipids identified to date, ∼4000 of which are sphingolipids. 29632068 10 53 theme 11-residue 1720:1729 arg1 region 1731:1736 an 11-residue region 1717:1736 an 11-residue region that determines the acyl-CoA specificity of CerS 1717:1785 In summary, we have identified an 11-residue region that determines the acyl-CoA specificity of CerS. 29632068 1 54 theme different 133:141 arg1 lipids 143:148 ∼40,000 different lipids 125:148 ∼40,000 different lipids 125:148 Lipids display large structural complexity, with ∼40,000 different lipids identified to date, ∼4000 of which are sphingolipids. 29632068 0 55 theme synthases 65:73 arg1 specificity 41:51 the acyl chain specificity 26:51 the acyl chain specificity of ceramide synthases 26:73 Eleven residues determine the acyl chain specificity of ceramide synthases. 29632068 7 56 theme significant 1262:1272 arg1 activity 1274:1281 significant activity 1262:1281 significant activity toward C24:1-CoA 1262:1298 Moreover, a chimeric protein, CerS4(291-301→CerS2), based on CerS4 (which normally generates C18-C22 ceramides) displayed significant activity toward C24:1-CoA. 29632068 6 57 from CerS2 1033:1037 arg1 residues 1019:1026 11 residues 1016:1026 11 residues from CerS2 (which generates C22-C24-ceramides) 1016:1073 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 1 58 theme lipids 143:148 arg1 sphingolipids 189:201 sphingolipids 189:201 sphingolipids 189:201 Lipids display large structural complexity, with ∼40,000 different lipids identified to date, ∼4000 of which are sphingolipids. 29632068 1 58 theme lipids 143:148 arg1 ∼4000 170:174 ∼4000 170:174 ∼4000 170:174 Lipids display large structural complexity, with ∼40,000 different lipids identified to date, ∼4000 of which are sphingolipids. 29632068 1 58 theme lipids 143:148 arg1 lipids 143:148 ∼40,000 different lipids 125:148 ∼40,000 different lipids 125:148 Lipids display large structural complexity, with ∼40,000 different lipids identified to date, ∼4000 of which are sphingolipids. 29632068 9 59 theme short 1481:1485 arg1 loop 1487:1490 this short loop 1476:1490 this short loop 1476:1490 Our findings may suggest that this short loop may restrict adjacent TMDs, leading to a more open conformation in the membrane, and that the CerS acting on shorter acyl-CoAs may have a longer, more flexible loop, permitting TMD flexibility. 29632068 0 60 theme ceramide 56:63 arg1 synthases 65:73 ceramide synthases 56:73 ceramide synthases 56:73 Eleven residues determine the acyl chain specificity of ceramide synthases. 29632068 6 61 contain containing 1005:1014 arg2 residues 1019:1026 11 residues 1016:1026 11 residues from CerS2 (which generates C22-C24-ceramides) 1016:1073 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 6 61 contain containing 1005:1014 arg1 CerS5 963:967 CerS5 963:967 CerS5 963:967 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 29632068 2 62 theme biological 238:247 arg1 activities 249:258 the biological activities 234:258 the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids 234:323 A critical factor determining the biological activities of the sphingolipid, ceramide, and of more complex sphingolipids is their N-acyl chain length, which in mammals is determined by a family of six ceramide synthases (CerS). 29632068 5 63 theme CerS 871:874 arg1 TMDs 858:861 TMDs 858:861 TMDs 858:861 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 63 theme CerS 871:874 arg1 domains 849:855 the last two putative transmembrane domains 813:855 the last two putative transmembrane domains (TMDs) of the CerS 813:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 5 63 theme CerS 871:874 arg1 CerS 871:874 the CerS 867:874 the CerS 867:874 Using site-directed mutagenesis and biochemical analyses, we now narrow specificity down to an 11-residue sequence in a loop located between the last two putative transmembrane domains (TMDs) of the CerS. 29632068 6 64 theme CerS5 963:967 arg1 backbone 951:958 the backbone 947:958 the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides) 947:1073 The specificity of a chimeric protein, CerS5(299-309→CerS2), based on the backbone of CerS5 (which generates C16-ceramide), but containing 11 residues from CerS2 (which generates C22-C24-ceramides), was altered such that it generated C22-C24 and other ceramides. 19561611 1 0 contain have 128:131 arg1 morphogens 117:126 Hedgehog (Hh) morphogens 103:126 Hedgehog (Hh) morphogens 103:126 Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. 19561611 1 0 contain have 128:131 arg2 roles 145:149 fundamental roles 133:149 fundamental roles 133:149 Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. 19561611 7 1 theme signaling 1178:1186 arg1 regulation 1161:1170 regulation 1161:1170 regulation of Hh signaling 1161:1186 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 19561611 6 2 theme Hh-HHIP 937:943 arg1 interface 945:953 the Hh-HHIP interface 933:953 the Hh-HHIP interface 933:953 Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. 19561611 4 3 theme HHIP 500:503 arg1 ectodomain 505:514 the human HHIP ectodomain 490:514 the human HHIP ectodomain 490:514 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 4 theme Desert 520:525 arg1 DHH 537:539 DHH 537:539 DHH 537:539 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 4 theme Desert 520:525 arg1 hedgehog 527:534 Desert hedgehog 520:534 Desert hedgehog (DHH) 520:540 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 3 5 dep conserved 382:390 arg1 vertebrate-specific 393:411 vertebrate-specific 393:411 vertebrate-specific 393:411 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 7 6 theme tuneable 1138:1145 arg1 mechanism 1147:1155 a tuneable mechanism 1136:1155 a tuneable mechanism for regulation of Hh signaling 1136:1186 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 19561611 4 7 from HHIP 567:570 arg1 complex 575:581 complex 575:581 complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+ 575:660 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 2 8 theme Hh 305:306 arg1 signals 308:314 Hh signals 305:314 Hh signals 305:314 Multiple cell-surface receptors are responsible for transducing and/or regulating Hh signals. 19561611 5 9 theme interaction 667:677 arg1 determinants 679:690 The interaction determinants 663:690 The interaction determinants 663:690 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 4 10 theme structures 475:484 arg1 DHH 537:539 DHH 537:539 DHH 537:539 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 10 theme structures 475:484 arg1 series 457:462 a series 455:462 a series of crystal structures for the human HHIP ectodomain 455:514 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 10 theme structures 475:484 arg1 hedgehog 527:534 Desert hedgehog 520:534 Desert hedgehog (DHH) 520:540 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 5 11 theme Hh 808:809 arg1 Zn2+ 811:814 the Hh Zn2+ and Ca2+ binding sites--functions 804:848 Zn2+ 811:814 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 4 12 theme human 494:498 arg1 ectodomain 505:514 the human HHIP ectodomain 490:514 the human HHIP ectodomain 490:514 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 7 13 theme sites 1121:1125 arg1 interplay 1086:1094 This interplay 1081:1094 This interplay of several metal binding sites 1081:1125 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 19561611 7 14 theme binding 1113:1119 arg1 sites 1121:1125 several metal binding sites 1099:1125 several metal binding sites 1099:1125 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 19561611 0 15 theme protein 89:95 arg1 HHIP 97:100 the human hedgehog-interacting protein HHIP 58:100 the human hedgehog-interacting protein HHIP 58:100 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 19561611 5 16 theme vertebrate 876:885 arg1 proteins 890:897 all vertebrate Hh proteins 872:897 all vertebrate Hh proteins 872:897 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 0 17 theme Structural 0:9 arg1 insights 11:18 Structural insights 0:18 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 0:101 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 19561611 6 18 theme key 913:915 arg1 contribution 917:928 a key contribution 911:928 a key contribution to the Hh-HHIP interface 911:953 Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. 19561611 1 19 theme Hedgehog 103:110 arg1 morphogens 117:126 Hedgehog (Hh) morphogens 103:126 Hedgehog (Hh) morphogens 103:126 Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. 19561611 1 20 theme Hh 192:193 arg1 signaling 195:203 Hh signaling 192:203 Hh signaling 192:203 Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. 19561611 0 21 theme hedgehog 25:32 arg1 sequestration 41:53 hedgehog ligand sequestration 25:53 hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP 25:100 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 19561611 1 22 theme signaling 195:203 arg1 dysregulation 175:187 dysregulation 175:187 dysregulation of Hh signaling 175:203 Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. 19561611 3 23 theme conserved 382:390 arg1 protein 355:361 the Hedgehog-interacting protein 330:361 the Hedgehog-interacting protein (Hhip) 330:368 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 3 23 theme conserved 382:390 arg1 inhibitor 413:421 a highly conserved, vertebrate-specific inhibitor 373:421 a highly conserved, vertebrate-specific inhibitor of Hh signaling 373:437 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 5 24 theme Hh 887:888 arg1 proteins 890:897 all vertebrate Hh proteins 872:897 all vertebrate Hh proteins 872:897 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 5 25 theme uncharacterized 761:775 arg1 functions 790:798 previously uncharacterized and distinct functions 750:798 previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins 750:897 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 3 26 theme Hh 426:427 arg1 signaling 429:437 Hh signaling 426:437 Hh signaling 426:437 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 5 27 theme Ca2+ 820:823 arg1 sites--functions 833:848 the Hh Zn2+ and Ca2+ binding sites--functions 804:848 sites--functions 833:848 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 4 28 theme Sonic 607:611 arg1 Shh 623:625 Shh 623:625 Shh 623:625 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 28 theme Sonic 607:611 arg1 HHIP-Shh 629:636 HHIP-Shh 629:636 HHIP-Shh 629:636 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 28 theme Sonic 607:611 arg1 hedgehog 613:620 Sonic hedgehog 607:620 Sonic hedgehog (Shh) (HHIP-Shh) 607:637 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 0 29 theme ligand 34:39 arg1 sequestration 41:53 hedgehog ligand sequestration 25:53 hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP 25:100 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 19561611 3 30 theme signaling 429:437 arg1 protein 355:361 the Hedgehog-interacting protein 330:361 the Hedgehog-interacting protein (Hhip) 330:368 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 3 30 theme signaling 429:437 arg1 inhibitor 413:421 a highly conserved, vertebrate-specific inhibitor 373:421 a highly conserved, vertebrate-specific inhibitor of Hh signaling 373:437 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 4 31 theme crystal 467:473 arg1 structures 475:484 crystal structures 467:484 crystal structures for the human HHIP ectodomain 467:514 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 2 32 theme cell-surface 232:243 arg1 receptors 245:253 Multiple cell-surface receptors 223:253 Multiple cell-surface receptors 223:253 Multiple cell-surface receptors are responsible for transducing and/or regulating Hh signals. 19561611 4 33 from isolation 545:553 arg1 complex 575:581 complex 575:581 complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+ 575:660 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 7 34 theme Hh 1175:1176 arg1 signaling 1178:1186 Hh signaling 1175:1186 Hh signaling 1175:1186 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 19561611 2 35 theme Multiple 223:230 arg1 receptors 245:253 Multiple cell-surface receptors 223:253 Multiple cell-surface receptors 223:253 Multiple cell-surface receptors are responsible for transducing and/or regulating Hh signals. 19561611 6 36 dep makes 905:909 arg1 whereas 956:962 whereas 956:962 whereas 956:962 Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. 19561611 4 37 with complex 575:581 arg1 HHIP-DHH 593:600 HHIP-DHH 593:600 HHIP-DHH 593:600 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 4 37 with complex 575:581 arg1 DHH 588:590 DHH 588:590 DHH (HHIP-DHH) 588:601 We have solved a series of crystal structures for the human HHIP ectodomain and Desert hedgehog (DHH) in isolation, as well as HHIP in complex with DHH (HHIP-DHH) and Sonic hedgehog (Shh) (HHIP-Shh), with and without Ca2+. 19561611 6 38 theme modulatory 1064:1073 arg1 role 1075:1078 an important modulatory role 1051:1078 an important modulatory role 1051:1078 Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. 19561611 7 39 theme several 1099:1105 arg1 sites 1121:1125 several metal binding sites 1099:1125 several metal binding sites 1099:1125 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 19561611 6 40 theme important 1054:1062 arg1 role 1075:1078 an important modulatory role 1051:1078 an important modulatory role 1051:1078 Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. 19561611 6 41 theme electrostatic 990:1002 arg1 repulsion 1004:1012 electrostatic repulsion 990:1012 electrostatic repulsion between the two proteins 990:1037 Zn2+ makes a key contribution to the Hh-HHIP interface, whereas Ca2+ is likely to prevent electrostatic repulsion between the two proteins, suggesting an important modulatory role. 19561611 5 42 theme biophysical 706:716 arg1 studies 718:724 biophysical studies 706:724 biophysical studies 706:724 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 1 43 theme fundamental 133:143 arg1 roles 145:149 fundamental roles 133:149 fundamental roles 133:149 Hedgehog (Hh) morphogens have fundamental roles in development, whereas dysregulation of Hh signaling leads to disease. 19561611 0 44 theme hedgehog-interacting 68:87 arg1 HHIP 97:100 the human hedgehog-interacting protein HHIP 58:100 the human hedgehog-interacting protein HHIP 58:100 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 19561611 3 45 theme Hedgehog-interacting 334:353 arg1 protein 355:361 the Hedgehog-interacting protein 330:361 the Hedgehog-interacting protein (Hhip) 330:368 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 3 45 theme Hedgehog-interacting 334:353 arg1 Hhip 364:367 Hhip 364:367 Hhip 364:367 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 3 45 theme Hedgehog-interacting 334:353 arg1 inhibitor 413:421 a highly conserved, vertebrate-specific inhibitor 373:421 a highly conserved, vertebrate-specific inhibitor of Hh signaling 373:437 Among these, the Hedgehog-interacting protein (Hhip) is a highly conserved, vertebrate-specific inhibitor of Hh signaling. 19561611 5 46 theme distinct 781:788 arg1 functions 790:798 previously uncharacterized and distinct functions 750:798 previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins 750:897 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 5 47 theme binding 825:831 arg1 sites--functions 833:848 the Hh Zn2+ and Ca2+ binding sites--functions 804:848 sites--functions 833:848 The interaction determinants, confirmed by biophysical studies and mutagenesis, reveal previously uncharacterized and distinct functions for the Hh Zn2+ and Ca2+ binding sites--functions that may be common to all vertebrate Hh proteins. 19561611 0 48 theme human 62:66 arg1 HHIP 97:100 the human hedgehog-interacting protein HHIP 58:100 the human hedgehog-interacting protein HHIP 58:100 Structural insights into hedgehog ligand sequestration by the human hedgehog-interacting protein HHIP. 19561611 7 49 theme metal 1107:1111 arg1 sites 1121:1125 several metal binding sites 1099:1125 several metal binding sites 1099:1125 This interplay of several metal binding sites suggests a tuneable mechanism for regulation of Hh signaling. 11726493 0 0 theme exclusion 57:65 arg1 domain 67:72 exclusion domain 57:72 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework 0:108 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 1 theme inflammatory 297:308 arg1 cells 310:314 immune and inflammatory cells 286:314 immune and inflammatory cells vital for defense of an organism 286:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 0 2 theme serine 157:162 arg1 proteases 164:172 granular serine proteases 148:172 granular serine proteases 148:172 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 3 3 theme proteins 600:607 arg1 approach 588:595 approach 588:595 approach of proteins 588:607 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 4 theme substrate 857:865 arg1 group 844:848 the N-terminal amino group 823:848 the N-terminal amino group of the substrate 823:865 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 4 theme substrate 857:865 arg1 substrate 857:865 the substrate 853:865 the substrate 853:865 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 2 5 theme endopeptidase 447:459 arg1 framework 420:428 the framework 416:428 the framework of a papain-like endopeptidase 416:459 The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. 11726493 3 6 theme exclusion 656:664 arg1 domain 666:671 the exclusion domain 652:671 the exclusion domain fastened within the tetrahedral framework 652:713 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 0 7 theme granular 148:155 arg1 proteases 164:172 granular serine proteases 148:172 granular serine proteases 148:172 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 3 8 theme Asp1 808:811 arg1 group 799:803 the carboxylic group 784:803 the carboxylic group of Asp1 784:811 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 8 theme Asp1 808:811 arg1 Asp1 808:811 Asp1 808:811 Asp1 808:811 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 4 9 theme structural 879:888 arg1 comparison 890:899 a structural comparison 877:899 a structural comparison 877:899 Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. 11726493 1 10 theme vital 316:320 arg1 cells 310:314 immune and inflammatory cells 286:314 immune and inflammatory cells vital for defense of an organism 286:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 5 11 dep fold 1183:1186 arg1 the 1179:1181 the 1179:1181 the 1179:1181 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 3 12 theme carboxylic 788:797 arg1 group 799:803 the carboxylic group 784:803 the carboxylic group of Asp1 784:811 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 12 theme carboxylic 788:797 arg1 Asp1 808:811 Asp1 808:811 Asp1 808:811 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 0 13 theme proteases 164:172 arg1 activation 134:143 activation 134:143 activation of granular serine proteases 134:172 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 14 theme physiological 227:239 arg1 activator 241:249 the physiological activator 223:249 the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism 223:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 14 theme physiological 227:239 arg1 I 196:196 Dipeptidyl peptidase I 175:196 Dipeptidyl peptidase I (DPPI) 175:203 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 14 theme physiological 227:239 arg1 C 218:218 cathepsin C 208:218 cathepsin C 208:218 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 2 15 theme protease-processing 486:504 arg1 enzyme 506:511 a robust oligomeric protease-processing enzyme 466:511 a robust oligomeric protease-processing enzyme 466:511 The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. 11726493 1 16 from groups 254:259 arg1 cells 310:314 immune and inflammatory cells 286:314 immune and inflammatory cells vital for defense of an organism 286:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 4 17 theme metallo-protease 1008:1023 arg1 inhibitor 1025:1033 a metallo-protease inhibitor 1006:1033 a metallo-protease inhibitor 1006:1033 Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. 11726493 2 18 theme oligomeric 475:484 arg1 enzyme 506:511 a robust oligomeric protease-processing enzyme 466:511 a robust oligomeric protease-processing enzyme 466:511 The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. 11726493 2 19 theme papain-like 435:445 arg1 endopeptidase 447:459 a papain-like endopeptidase 433:459 a papain-like endopeptidase 433:459 The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. 11726493 3 20 theme tetrahedral 518:528 arg1 arrangement 530:540 The tetrahedral arrangement 514:540 The tetrahedral arrangement of the active sites exposed to solvent 514:579 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 4 21 theme site 936:939 arg1 cleft 941:945 the active site cleft 925:945 the active site cleft 925:945 Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. 11726493 3 22 theme tetrahedral 693:703 arg1 framework 705:713 the tetrahedral framework 689:713 the tetrahedral framework 689:713 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 23 theme sites 556:560 arg1 arrangement 530:540 The tetrahedral arrangement 514:540 The tetrahedral arrangement of the active sites exposed to solvent 514:579 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 5 24 theme Haim-Munk 1111:1119 arg1 syndromes 1142:1150 Haim-Munk and Papillon-Lefevre syndromes 1111:1150 Haim-Munk and Papillon-Lefevre syndromes 1111:1150 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 0 25 theme dipeptidyl 19:28 arg1 C 53:53 cathepsin C 43:53 cathepsin C 43:53 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 0 25 theme dipeptidyl 19:28 arg1 peptidase 30:38 human dipeptidyl peptidase I 13:40 human dipeptidyl peptidase I (cathepsin C) 13:54 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 3 26 theme polypeptide 738:748 arg1 chain 750:754 a polypeptide chain 736:754 a polypeptide chain 736:754 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 1 27 theme Dipeptidyl 175:184 arg1 DPPI 199:202 DPPI 199:202 DPPI 199:202 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 27 theme Dipeptidyl 175:184 arg1 I 196:196 Dipeptidyl peptidase I 175:196 Dipeptidyl peptidase I (DPPI) 175:203 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 27 theme Dipeptidyl 175:184 arg1 activator 241:249 the physiological activator 223:249 the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism 223:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 27 theme Dipeptidyl 175:184 arg1 C 218:218 cathepsin C 208:218 cathepsin C 208:218 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 0 28 theme human 13:17 arg1 C 53:53 cathepsin C 43:53 cathepsin C 43:53 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 0 28 theme human 13:17 arg1 peptidase 30:38 human dipeptidyl peptidase I 13:40 human dipeptidyl peptidase I (cathepsin C) 13:54 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 29 theme groups 254:259 arg1 activator 241:249 the physiological activator 223:249 the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism 223:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 29 theme groups 254:259 arg1 I 196:196 Dipeptidyl peptidase I 175:196 Dipeptidyl peptidase I (DPPI) 175:203 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 29 theme groups 254:259 arg1 C 218:218 cathepsin C 208:218 cathepsin C 208:218 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 0 30 theme endopeptidase 86:98 arg1 framework 100:108 an endopeptidase framework 83:108 an endopeptidase framework 83:108 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 31 theme organism 340:347 arg1 defense 326:332 defense 326:332 defense of an organism 326:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 3 32 theme active 549:554 arg1 sites 556:560 the active sites 545:560 the active sites exposed to solvent 545:579 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 1 33 theme peptidase 186:194 arg1 DPPI 199:202 DPPI 199:202 DPPI 199:202 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 33 theme peptidase 186:194 arg1 I 196:196 Dipeptidyl peptidase I 175:196 Dipeptidyl peptidase I (DPPI) 175:203 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 33 theme peptidase 186:194 arg1 activator 241:249 the physiological activator 223:249 the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism 223:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 33 theme peptidase 186:194 arg1 C 218:218 cathepsin C 208:218 cathepsin C 208:218 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 34 from cells 310:314 arg1 groups 254:259 groups 254:259 groups of serine proteases from immune and inflammatory cells vital for defense of an organism 254:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 34 from cells 310:314 arg1 proteases 271:279 serine proteases 264:279 serine proteases from immune and inflammatory cells vital for defense of an organism 264:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 0 35 theme peptidase 30:38 arg1 Structure 0:8 Structure 0:8 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework 0:108 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 36 theme serine 264:269 arg1 proteases 271:279 serine proteases 264:279 serine proteases from immune and inflammatory cells vital for defense of an organism 264:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 3 37 theme native 618:623 arg1 state 625:629 their native state 612:629 their native state 612:629 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 4 38 theme active 929:934 arg1 cleft 941:945 the active site cleft 925:945 the active site cleft 925:945 Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. 11726493 0 39 theme cathepsin 43:51 arg1 C 53:53 cathepsin C 43:53 cathepsin C 43:53 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 0 39 theme cathepsin 43:51 arg1 peptidase 30:38 human dipeptidyl peptidase I 13:40 human dipeptidyl peptidase I (cathepsin C) 13:54 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 40 theme proteases 271:279 arg1 groups 254:259 groups 254:259 groups of serine proteases from immune and inflammatory cells vital for defense of an organism 254:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 40 theme proteases 271:279 arg1 proteases 271:279 serine proteases 264:279 serine proteases from immune and inflammatory cells vital for defense of an organism 264:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 5 41 theme mutations 1061:1069 arg1 location 1040:1047 The location 1036:1047 The location 1036:1047 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 3 42 theme domain 666:671 arg1 body 644:647 the massive body 632:647 the massive body of the exclusion domain fastened within the tetrahedral framework 632:713 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 43 theme chain 750:754 arg1 approach 724:731 approach 724:731 approach of a polypeptide chain 724:754 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 5 44 theme Papillon-Lefevre 1125:1140 arg1 syndromes 1142:1150 Haim-Munk and Papillon-Lefevre syndromes 1111:1150 Haim-Munk and Papillon-Lefevre syndromes 1111:1150 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 5 45 theme missense 1052:1059 arg1 mutations 1061:1069 missense mutations 1052:1069 missense mutations 1052:1069 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 3 46 theme N-terminal 827:836 arg1 group 844:848 the N-terminal amino group 823:848 the N-terminal amino group of the substrate 823:865 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 46 theme N-terminal 827:836 arg1 substrate 857:865 the substrate 853:865 the substrate 853:865 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 1 47 theme immune 286:291 arg1 cells 310:314 immune and inflammatory cells 286:314 immune and inflammatory cells vital for defense of an organism 286:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 3 48 theme amino 838:842 arg1 group 844:848 the N-terminal amino group 823:848 the N-terminal amino group of the substrate 823:865 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 48 theme amino 838:842 arg1 substrate 857:865 the substrate 853:865 the substrate 853:865 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 3 49 theme massive 636:642 arg1 body 644:647 the massive body 632:647 the massive body of the exclusion domain fastened within the tetrahedral framework 632:713 The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. 11726493 5 50 theme enzyme 1208:1213 arg1 function 1192:1199 function 1192:1199 function 1192:1199 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 5 50 theme enzyme 1208:1213 arg1 fold 1183:1186 fold 1183:1186 fold 1183:1186 The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. 11726493 2 51 theme robust 468:473 arg1 enzyme 506:511 a robust oligomeric protease-processing enzyme 466:511 a robust oligomeric protease-processing enzyme 466:511 The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. 11726493 2 52 theme additional 387:396 arg1 domain 398:403 an additional domain 384:403 an additional domain 384:403 The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. 11726493 4 53 theme exclusion 973:981 arg1 domain 983:988 the exclusion domain 969:988 the exclusion domain 969:988 Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. 11726493 0 54 dep Structure 0:8 arg1 domain 67:72 exclusion domain 57:72 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework 0:108 Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. 11726493 1 55 theme cathepsin 208:216 arg1 activator 241:249 the physiological activator 223:249 the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism 223:347 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 55 theme cathepsin 208:216 arg1 I 196:196 Dipeptidyl peptidase I 175:196 Dipeptidyl peptidase I (DPPI) 175:203 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. 11726493 1 55 theme cathepsin 208:216 arg1 C 218:218 cathepsin C 208:218 cathepsin C 208:218 Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism.